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Permit applications 08-011-106rm 

and 08-014-101rm received from 

ArborGen LLC

Field testing of genetically engineered 

Eucalyptus grandis X Eucalyptus 

urophylla

Final Environmental Assessment

April 2010

Biotechnology Regulatory Services 

Agency Contact

Cynthia Eck, Document Control Officer 

USDA, APHIS, BRS 

4700 River Road, Unit 147 

Riverdale, MD 20737-1237 

Phone: (301) 734-0667

Fax: (301) 734-8669 

Cynthia.A.Eck@aphis.usda.gov 

____________________________________

The U.S. Department of Agriculture (USDA) prohibits 

discrimination in all its programs and activities on the basis of race, 

color, national origin, sex, religion, age, disability, political beliefs, 

sexual orientation, or marital or family status. (Not all prohibited 

bases apply to all programs.) Persons with disabilities who require 

alternative means for communication of program information 

(Braille, large print, audiotape, etc.) should contact USDA’S 

TARGET Center at (202) 720–2600 (voice and TDD).

United States

Department of 

Agriculture

Marketing and 

Regulatory 

Programs

Animal and 

Plant Health 

Inspection 

Service
Page 2 of 98

To file a complaint of discrimination, write USDA, Director, Office of 

Civil Rights, Room 326–W, Whitten Building, 1400 Independence 

Avenue, SW, Washington, DC 20250–9410 or call (202) 720–5964 

(voice and TDD). USDA is an equal opportunity provider and 

employer.

____________________________________

Mention of companies or commercial products in this report does not 

imply recommendation or endorsement by the U.S. Department of 

Agriculture over others not mentioned. USDA neither guarantees 

nor warrants the standard of any product mentioned. Product names 

are mentioned solely to report factually on available data and to 

provide specific information.

____________________________________

This publication reports research involving pesticides. All uses of 

pesticides must be registered by appropriate State and/or Federal 

agencies before they can be recommended. 

____________________________________

CAUTION: Pesticides can be injurious to humans, domestic animals, 

desirable plants, and fish or other wildlife—if they are not handled or applied 

properly. Use all pesticides selectively and carefully. Follow recommended

practices for the disposal of surplus pesticides and pesticide containers.
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TABLE OF CONTENTS

I. SUMMARY................................................................................................................................... 4

II. PURPOSE AND NEED.............................................................................................................. 5

A. PROPOSED ACTION................................................................................................................... 5

B. PURPOSE OF THIS ENVIRONMENTAL ASSESSMENT................................................................ 5

C. NEED FOR THIS ACTION........................................................................................................... 6

D. PURPOSE AND DESCRIPTION OF THE RESEARCH ................................................................... 6

III. AFFECTED ENVIRONMENT ................................................................................................ 6

IV. ALTERNATIVES.................................................................................................................... 16

A. NO ACTION – DENY THE PERMIT .......................................................................................... 17

B. PREFERRED ALTERNATIVE – ISSUE THE APHIS PERMIT.................................................... 17

V. ENVIRONMENTAL CONSEQUENCES.............................................................................. 18

A. NO ACTION.............................................................................................................................. 18

B. PREFERRED ALTERNATIVE.................................................................................................... 18

C. CUMULATIVE EFFECTS .......................................................................................................... 33

D. THE DEGREE TO WHICH THE POSSIBLE EFFECTS ARE HIGHLY UNCERTAIN OR INVOLVE 

UNIQUE OR UNKNOWN RISKS ........................................................................................ 34

E. RISKS TO THREATENED AND ENDANGERED SPECIES .......................................................... 37

F. OTHER CONSIDERATIONS ...................................................................................................... 38

G. CONCLUSION........................................................................................................................... 39

VI. LISTING OF AGENCIES AND PERSONS CONSULTED............................................... 40

VII. REFERENCES....................................................................................................................... 40

APPENDIX I: STATUS OF EXISTING FIELD TESTS ALLOWED TO FLOWER ............ 46

APPENDIX II: HYDROLOGY CONSIDERATIONS FOR PLANTED 

EUCALYPTUS SUBMITTED BY ARBORGEN LLC.................................................... 48

APPENDIX III: USDA FOREST SERVICE ASSESSMENT OF IMPACTS ON 

HYDROLOGY.................................................................................................................... 59

APPENDIX IV: THREATENED AND ENDANGERED SPECIES ANALYSIS.................... 68

APPENDIX V: SUPPLEMENTAL PERMIT CONDITIONS................................................... 94
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I. Summary

United States Department of Agriculture (USDA), Animal Plant Health Inspection 

Service (APHIS) has prepared an environmental assessment (EA) in response to confined 

environmental release permit applications (APHIS Number 08-011-106rm and 08-014-

101rm) received from ArborGen LLC (ArborGen) to continue research on genetically 

engineered (transgenic) Eucalyptus trees currently permitted by APHIS, to allow the 

planting of additional trees, and to allow genetically engineered (GE) Eucalyptus trees to 

flower at confined field site locations. These plants are a clone1 coded EH1 derived from 

a hybrid of Eucalyptus grandis X Eucalyptus urophylla and have been genetically 

engineered with different constructs. The purpose of the confined environmental release 

is for ArborGen to assess the effectiveness of gene constructs which are intended to 

confer cold tolerance; to test the efficacy of a gene introduced to alter lignin biosynthesis; 

and to test the efficacy of a gene designed to alter fertility. In addition the trees have 

been engineered with a selectable marker gene which confers resistance to the antibiotic 

kanamycin.

ArborGen has been granted several permits for transgenic Eucalyptus trees, some of 

which allow flowering and some of which do not allow flowering. ArborGen previously 

applied for a permit (06-325-111r) requesting that transgenic Eucalyptus trees be allowed 

to flower at a confined field site in Alabama on 1.1 acres. The permit was granted 

following the completion of an EA and reaching of a FONSI 

(http://www.aphis.usda.gov/brs/aphisdocs/06_325111r_ea.pdf). This permit was 

subsequently amended to allow flowering on an additional 5.1 acres at the same location 

bringing the total acres at this site allowed to flower to 6.2 acres. ArborGen has also 

been granted permit 08-151-101r to allow flowering of transgenic Eucalyptus on 1.4 

acres on a confined field trial site in Florida. Under permits 06-325-111r and 08-151-

101r, a total of 7.6 acres of trees at 2 sites are authorized by APHIS to flower. These two 

permits were issued for three years and will expire on June 27, 2010 and June 30, 

2011repectively. ArborGen has been allowed to plant trees under a different permit 08-

039-102rm, but they cannot flower under this permit which expires April 25, 2011. All 

multiyear field trials are issued under three-year permits, and if applicants wish to 

continue the field trials thay can renew these permits for an additional 3 years.

 

ArborGen has submitted two new permit applications 08-014-101rm and 08-011-106rm, 

for planting and growing trees on 28 sites and is also requesting that trees on 27 of these 

sites be allowed to flower (the subject of this EA). The original request was to plant 29 

sites and allow 28 to flower, however, one location was removed from permit application 

08-014-101rm, which reduced the number to 28 sites, with 27 allowed to flower. The 28 

sites include those already authorized for planting under permit 08-039-102rm, As of 

April 2010, ArborGen is currently growing approximately 37 acres of the transgenic 

Eucalyptus hybrid on 15 sites under APHIS permit 08-039-102rm that would be allowed 

to flower under the two new permits if approved. These two new permits (08-014-101rm 

and 08-011-106rm) combined would allow flowering on up to 330 acres across 27 

 1 Clone – as defined in horticulture and forestry means is a population of genetically identical plants that 

has been derived from one individual. Despite popular use of the word, a clone is not an individual.
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locations. The size of each confined field test site in these new permit applications 

ranges from 0.5 to 20 acres. 

The genes are the same as those in permits 06-325-111r and 08-151-101r, with the 

addition of a gene aimed at altering lignin biosynthesis that is being tested on some of the 

sites. The status of the field tests permitted under permits 06-325-111r and 08-151-101r 

is included in Appendix I.

II. Purpose and Need

A. Proposed Action

The proposed action is for APHIS, Biotechnology Regulatory Services (BRS) to issue 

two confined environmental release permits to allow the planting, field testing and 

flowering of a GE Eucalyptus hybrid clone engineered to express various genes on 28

confined field site locations in the southeastern United States. The genes are intended to 

confer increased tolerance to cold temperatures, alter fertility and alter lignin

biosynthesis. In addition there is a gene used as a selectable marker that confers no other 

benefit to the GE Eucalyptus trees.

B. Purpose of this Environmental Assessment

The purpose of this EA is to assess potential environmental impacts of field research 

studies being conducted on 28 research sites in Alabama, Florida, Georgia, Louisiana, 

Mississippi, South Carolina, and Texas. Two permit applications were received by 

APHIS–BRS in January 2008 from ArborGen to continue research on GE Eucalyptus

hybrid trees originally planted under APHIS authorized notifications, to plant additional 

trees, and to allow all the trees except the trees on one of the sites in South Carolina to 

flower. These are permit application numbers 08-011-106rm and 08-014-101rm.

This EA was conducted pursuant to: (1) The National Environmental Policy Act of 1969 

(NEPA), as amended (42 U.S.C. § 4321 et seq.), (2) regulations of the Council on 

Environmental Quality for implementing the procedural provisions of NEPA (40 CFR 

parts 1500-1508), (3) USDA regulations implementing NEPA (7 CFR part 1b), and (4) 

APHIS' NEPA Implementing Procedures (7 CFR part 372). Generally, issuances of 

permits for confined field trials of regulated articles are categorically excluded under 

APHIS NEPA Implementing Procedures. However, if APHIS determines that a confined 

field release of genetically engineered organisms may have the potential to significantly 

impact the quality of the human environment, as those terms are defined in 40 CFR 

1508.27 and 1508.14, an environmental assessment may be prepared, pursuant to 7 CFR 

§ 372.5(d) to determine if an Environmental Impact Statement is warranted. In this case, 

an EA has been prepared because the permittee intends to allow GE Eucalyptus trees to 

grow under an APHIS confined environmental release permit for 7 years to allow the 

trees to reach maturity and flower. The actions described in the permit applications 

involve the confined field release of transgenic Eucalyptus grandis x Eucalyptus 

urophylla into the environment. APHIS has prepared this EA to address concerns that 

the increased number of locations and size of the releases that would be allowed to flower 

could potentially result in the unconfined release of pollen and/or seed into the 
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environment. Other impacts such as hydrology, allelopathy and fire are also addressed in 

this EA.

C. Need for This Action

USDA-APHIS-BRS mission is to protect America’s agriculture and the environment 

using a dynamic and science-based regulatory framework that allows for the safe 

development and use of GE organisms. APHIS’ regulations in 7 CFR part 340, which 

were promulgated pursuant to authority granted by the Plant Protection Act, as amended, 

(7 U.S.C. 7701–7772), regulates the introduction (importation, interstate movement, or 

release into the environment) of certain GE organisms and products. The Plant 

Protection Act directs the USDA to facilitate imports and interstate commerce in 

agricultural products in ways that will reduce, to the extent practicable, the risk of 

dissemination of plant pests. Under APHIS regulations, the APHIS Administrator has 

authority to regulate any organism or product altered or produced through genetic 

engineering that the Administrator determines is a plant pest or has reason to believe is a 

plant pest. When APHIS receives an application for a permit for environmental release, 

the application is evaluated to determine whether the environmental release, with 

appropriate conditions imposed, can be carried out while preventing the dissemination 

and establishment of plant pests. The receipt of a permit application to introduce a 

genetically engineered organism requires a response from the Administrator:

Administrative action on applications. After receipt and review by 

APHIS of the application and the data submitted pursuant to paragraph 

(a) of this section, including any additional information requested by 

APHIS, a permit shall be granted or denied (7 CFR 340.4(e)).

The applicant has provided the information associated with this request in the permit 

application. This information has been reviewed and analyzed in this EA. 

D. Purpose and Description of the Research

The purpose of the ArborGen research is to assess the efficacy of the introduced cold 

tolerance genes and gene to alter lignin biosynthesis in Eucalyptus. According to the 

applicant, genetically engineered cold tolerant Eucalyptus would enable the production of 

this hardwood species for pulping and for biofuel applications in managed plantation 

forests in the southeastern U.S. In addition, the applicant is researching mechanisms for 

altered fertility. The confined release of the trees in different areas of the southeast U.S. 

will allow the applicant to obtain data on performance of the transgenic trees and the 

efficacy of the inserted genes in a wide variety of environments.

III. Affected Environment

Biology of Eucalyptus and Status in the United States
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The genus Eucalyptus belongs to family Myrtaceae (subfamily: Leptospermoideae) 

which includes over 700 species. Eucalyptus is native to Australia with the exception of 

some species that are native to the Timor Islands (Groves 1994, Ladiges 1997). There 

are no wild relatives of Eucalyptus that occur naturally in the United States. An overview 

of the biology of Eucalyptus grandis has been published by the US Forest Service 

(Meskimen and Francis 1990). Eucalyptus has been planted as an ornamental species in 

the extreme southern United States where mild winters will allow some species to grow. 

Eucalyptus normally propagates in its native range via seeds. It does not spread 

vegetatively like other trees such as poplar or willow. In the United States it is usually 

propagated and sold commercially as rooted stem cuttings. 

There have been numerous attempts to grow Eucalyptus as a commercial forest tree in the 

southeastern United States, but due to its sensitivity to cold temperatures, these attempts 

have not met with success. It is only grown in commercial plantations in central and 

southern Florida, where it normally survives freezing temperatures. Eucalyptus is 

adapted to live in the mild arid and semi-arid climate of Australia. Severe freezing events 

that can occur in the southern United States have limited its establishment as a 

commercial forest tree. There are plantations of Eucalyptus grandis and E. amplifolia

currently grown in south central Florida as short rotation energy crops and for mulch 

production (Stricker et al. 2000, Rockwood et al. 2004). These trees are generally 

planted in areas where severe freezing events are rare.

The species hybrid E. grandis x E. urophylla that ArborGen wishes to allow to mature 

and flower under this permit has not been categorized as invasive. The E. grandis x E. 

urophylla hybrid has been grown for forty years in South America and during this time 

there has been no evidence of invasiveness by into natural forest areas which are growing 

as part of an integrated land management system (Luis Silva, International Paper 

Company, Brazil, comment to docket APHIS-2008-0059). One of the parents of the 

hybrid, E. grandis has been shown to be naturalized in the State of Florida

(http://www.florida.plantatlas.usf.edu). In South Africa, E. grandis has been shown to be 

an invasive species (Forsyth et al., 2004). It is also naturalized in both New Zealand 

(New Zealand Plant Conservation Network 2005: 

http://www.nzpcn.org.nz/exotic_plant_life_and_weeds/weed_list.asp.) and Ecuador

(http://i3n.iabin.net/participants/ecuador.html). Daehler (1998) listed E. grandis as an 

invader of natural areas. These and other data were recently used in the University of 

Florida’s IFAS Assessment of Non-native Plants in Florida’s Natural Areas. The 

assessment conclusion for E. grandis is that it is now: "Predicted to be invasive: 

recommend only under specific management practices that have been approved by the 

IFAS Invasive Plant 2 Working Group" 

(http://plants.ifas.ufl.edu/assessment/predictive_response_forms.html). Specific 

management practices for four specific cultivars of E. grandis have been approved by the 

Working Group (http://plants.ifas.ufl.edu/assessment/ -- see Approved, Specified and 

Limited Uses link). The University of Florida IFAS has not made an assessment of the E. 

grandis and E. urophylla hybrid. 
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Numerous species of Eucalyptus were introduced into California during that State’s early 

history (see Santos: http://wwwlibrary.csustan.edu/bsantos/euctoc.htm), and some of 

these species have become established. Two of these, E. globulus (Tasmanian blue gum) 

and E. camaldulensis (Red gum) are now categorized as invasive by the California 

Invasive Plant Council (http://www.cal-ipc.org/ip/inventory/weedlist.php). Neither of 

these species is being proposed to be planted at the permitted field site. 

Traits Engineered into Eucalyptus

ArborGen LLC wishes to field test genetically engineered (transgenic) Eucalyptus trees 

during which time the trees may flower. These plants are a clone coded EH1 derived 

from a hybrid of Eucalyptus grandis X Eucalyptus urophylla. These have been 

genetically engineered with different constructs. The purpose of the field trials is to test 

the effectiveness of the CBF gene which is intended to confer cold tolerance and to test 

the efficacy of the Barnase gene designed to alter fertility. In a small set of experiments 

the CBF and Barnase genes are also being tested in combination with genes introduced to 

alter lignin biosynthesis (claimed as CBI). In addition the trees have been engineered 

with a common selectable marker gene (nptII) which confers resistance to the antibiotic 

kanamycin.

Confined Field Test Locations

The confined field tests are taking place on land controlled by ArborGen or through 

contracts for field testing. The exact locations are claimed as Confidential Business 

Information (CBI) and have been submitted as part of the APHIS permit application. 

Under the two permits, there are 28 sites where trees have been planted or will be 

planted, and on 27 of these sites the trees will be allowed to flower. See below for the 

States and Counties in which these 28 sites are located. An additional site in South 

Carolina is a holding area for plants in pots and trees. Trees will be held there for 

planting and will not be allowed to flower.

All the confined field test sites listed in this permit application are either on privately 

owned managed plantation forests and agricultural farm lands or experimental research 

stations managed by academic institutions and industry. The standard agricultural and

silvicultural practices for land preparation, planting, irrigation, and harvesting of plants 

have been routinely used on these sites. Sites that include managed pastures have had 

intense activity including the use of heavy machinery for general upkeep, irrigation, 

fertilization, controlled grazing and management of grasses. Standard silvicultural

practices will be used at these sites for the duration of the field tests. Surveys conducted 

by the applicant at each of these locations indicate that there are not any old growth 

forests or undisturbed natural areas in the immediate surroundings of the test sites. The 

trees will be planted from 0.5 up to 20 acres, depending on the location. In the case of 

these tests the planting density will be from 300 - 600 trees per acre2

 2 Planting density typically refers to the number of trees per acre. Planting densities can vary greatly 

depending upon the tree species and the environment, but densities of short rotation hardwood trees in the 

southeastern US are typically in the range of 300–800 trees per acre. Therefore sites ranging from 10 to 20 

. An acre is about 

the size of a football field.
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Baldwin County Alabama Site:

This location has been an agricultural research station for more than 20 years. The location 

has been used for managed production of annual agricultural crops and forest trees. Site 

preparation involved herbicide application, subsoiling, and planting of trees in flat beds. The 

surrounding areas of the test site consist of field plantings of agricultural crops, experimental 

forest trees and an abandoned pecan orchard. Approximately 6.2 acres of existing field trials 

of genetically modified Eucalyptus of the same clone (EH1) are being grown under issued 

permit # 06-325-111r under which these trees are allowed to flower. The oldest of these trees 

at this site are now entering their fifth growing season.

Up to 8,000 containerized trees, including non-transgenic trees could be transferred to this 

field test site and planted in field plots of up to 10 acres (at around 300 - 600 trees per acre) 

over the next three years. An additional field trial of the same clone (EH1) was established at 

this site on approximately 0.8 acres now covered under permit 08-039-102rm. This trial 

would be transferred to permit # 08-011-106rm to allow flowering. 

Escambia County Alabama Site:

This location had previously been used as an intensely managed pasture for more than 5 

years and was planted with grasses suitable for cattle grazing. Site preparation involved 

herbicide application to remove existing grasses, subsoiling, preparation for possible 

irrigation, and planting of the test trees in flat beds. The surrounding areas of the test site 

consist of experimental forest trees; approximately 30 year-old slash pine and a re-forested 

area with less than 7 year-old mixed stands of pine and hardwood species.

Up to 8,000 containerized trees, including non-transgenic trees could be transferred to this 

field test site and planted in field plots of up to 10.3 acres over the next three years. The 

existing field trials of the same clone (EH1) were planted at this site on approximately 0.5 

acres that are now covered under Permit #08-039-102rm. These trials would be transferred to 

permit # 08-011-106rm to allow flowering. 

Evans County Georgia Site:

This location has been a commercial nursery for forest seedling production for over 30 years. 

Site preparation involved herbicide application, subsoiling and planting of trees in flat beds. 

The areas surrounding the test site consist of nursery beds of forest tree seedlings, 

experimental forest trees, agricultural crops and mixed stands of hardwood and pine. 

Up to 4,000 containerized trees, including non-transgenic trees, could be transferred to this 

field test site and planted on field plots of up to 5 acres over the next three years. A field trial 

of the same clone (EH1) was planted on approximately 0.2 acres at this site that is now 

covered under BRS permit # 08-039-102rm. This trial would be transferred to permit # 08-

011-106rm to allow flowering.

 acres can have from 3000 to 16,000 total trees planted in the ground. Twenty acres, as defined by forest 

plantation standards in the southeast, is considered a small planting.
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Saint Landry’s Parrish Louisiana Site:

This location has been an experimental agricultural farm for more than 25 years, used for 

conducting research experiments with soybean, cotton and wheat. Site preparation involved 

herbicide application, subsoiling and planting of trees in flat beds. The areas surrounding the 

test site consist of agricultural fields of rice, sugarcane and millet.

Up to 8,000 containerized trees, including non-transgenic trees could be transferred to this 

field test site and planted in field plots of up to 11.7 acres over the next three years. Field 

trials of the same clone (EH1) were planted at this site on approximately 1.9 acres under 

Permit #08-039-102rm. All tests except 0.2 acres were subsequently terminated. The 

remaining 0.2 acres would be transferred to permit # 08-011-106rm to allow flowering.

Marshall County Mississippi Site:

This location has been an agricultural research station for more than 50 years, used for 

conducting research experiments with agricultural crops and grasses. The test site was 

previously used for experimental planting of grasses. Site preparation involved herbicide 

application, subsoiling and planting of trees in flat beds. The areas surrounding the test site 

consist of experimental forest trees, agricultural fields, and less than 5 year-old pine 

plantations.

A field test of the same clone (EH1) was established at this site on 0.5 acres under permit 08-

039-102rm. This test was subsequently terminated. No additional plantings are currently 

anticipated at this site. Additional tests, if planted at this site, would be allowed to flower 

under permit # 08-011-106rm.

Pearl River County Mississippi Site:

This location has been used as an agricultural research station for more than 5 years for 

conducting research experiments with agricultural crops and grasses. The test site was used 

for experimental planting of grasses. Site preparation involved herbicide application to 

remove existing grasses, subsoiling, preparation for irrigation installation, and planting of 

trees in flat beds. The areas surrounding the test site consist of a grape research farm, mixed 

stands of hardwoods and pine, and a residential area.

Up to 4,000 containerized trees, including non-transgenic trees could be transferred to this 

field test site and planted in field plots of up to 7 acres over the next three years. The existing 

field trials of the same clone (EH1) were established at this site on approximately 3.0 acres 

that are now covered under permit 08-039-102rm. These trials would be transferred to 

permit # 08-011-106rm to allow flowering.

Bamberg County South Carolina Site:

This location has been a managed forest plantation for more than 14 years. The location has 

been specifically used for short-rotation planting of hardwoods and softwood trees for 

forestry research. Standard silvicultural practices for site preparation, irrigation, fertilization, 

planting and harvesting have been used at this location. Similar practices will be used for the 

additional field tests to be established at this site. The areas surrounding the test site consist 
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of experimental forest trees, young pine plantations, mixed stands of hardwoods and pine, 

and agricultural fields. There are experimental test plots of non-transgenic cold-hardy E. 

macarthurii, E. benthamii, E. viminalis, E. badjensis, and E. dorrigoensis planted at least 

1000 meters from the test plot location.

Up to 4,000 containerized trees, including non-transgenic trees could be transferred to this 

field test site and planted in field plots of up to 8.3 acres over the next three years. Field 

trials of the same clone (EH1) were established at this site on 3.5 acres under permit 08-039-

102rm. All tests except 0.2 acres were subsequently terminated. The remaining 0.2 acres 

would be transferred to permit # 08-011-106rm to allow flowering.

Berkeley County South Carolina Site 1:

This is an extension of a greenhouse facility that has been used for acclimatization of 

transgenic and non-transgenic plants for more than 7 years. The 0.5 acre release site is 

located adjacent to greenhouse facilities and is surrounded by hardwoods and pine 

plantations. This site is a secure fenced holding area where trees growing in containers 

are transferred from the greenhouse to the out-of-doors for acclimatization prior to field 

planting. Trees will not be allowed to flower at this location.

Berkeley County South Carolina Site 2:

This location has been a managed forest plantation for more than 7 years. The location 

has been specifically used for short rotation planting of cottonwood and Eucalyptus

Hybrid (EH1) for forestry research. Site preparation involved herbicide application, 

subsoiling, drip irrigation installation, and planting of trees in flat beds. The test site is 

located adjacent to greenhouse facilities and is surrounded by pine plantations.

Field trials of the same clone (EH1) were established at this site on 0.5 acres under permit 

08-039-102rm. These tests were subsequently terminated. No additional plantings are 

currently anticipated at this site. Additional tests, if planted at this site, would be allowed to 

flower under permit # 08-011-106rm.

Charleston County South Carolina Site: 

This location has been a managed forest plantation for more than 10 years. The location has 

been specifically used for short-term planting of hardwoods and softwood trees for forestry 

research. Standard silvicultural practices for site preparation, irrigation, fertilization, planting 

and harvesting were used at this location. Similar practices were used for the additional field 

tests established at this site. The areas adjacent to the field test site include young mixed 

stands of hardwoods and pines.

Field trials of the same clone (EH1) were established at this site on 3.1 acres under permit 08-

039-102rm. All tests were subsequently terminated. No additional trials are currently 

planned for this site. Additional tests, if planted at this site, would be allowed to flower under

permit # 08-011-106rm.

Marlboro County South Carolina Site:
Page 12 of 98

This location has been a commercial nursery for forest seedling production for over 30 years. 

Site preparation involved herbicide application, subsoiling and planting of trees in flat beds. 

The surrounding areas consist of field plantings of agricultural crops, nursery beds of forest 

tree seedlings and less than 30 year-old mixed hardwood and pine plantations.

A field trial of the same clone (EH1) was established at this site on 0.3 acre under permit 08-

039-102rm. This test was subsequently terminated. No additional trials are currently planned 

for this site. Additional tests, if planted at this site, would be allowed to flower under permit 

# 08-011-106rm.

Hardin County Texas Site:

This location has been a managed forest plantation for more than 30 years. The location 

consists of mixed hardwood tree plantations planted using standard silvicultural practices and 

was harvested by the owner in 2004. The test site is within the larger harvested area and was 

re-bedded by the owner for planting. Site preparation included herbicide application and subsoiling. The areas surrounding the test site consist of mixed hardwood stands and managed 

loblolly pine plantations. There are experimental test plots of non-transgenic cold-hardy 

Eucalyptus species, E. macarthurii, E. benthamii, and E. viminalis planted within 100 m of 

the transgenic test plot location.

Up to 16,000 containerized trees, including non-transgenic trees, could be transferred to this 

field test site and planted in field test plots of up to 20 acres over three years. A field trial of 

the same clone (EH1) was established at this site on 19.7 acres under permit 08-039-102rm.

This trial would be transferred to permit # 08-011-106rm to allow flowering.

Jasper County Texas Site 1:

This location has been a managed pine plantation for more than 25 years. Previous plantings 

were cultivated in beds using standard silvicultural practices. The existing pine plantation at 

this site was harvested by the owner in 2007 and re-bedded for planting. The test site is 

within the larger harvested and bedded site prepared by the site owner. Further site 

preparation involved herbicide application, subsoiling and planting of trees in flat beds. The 

areas surrounding the test site consist of harvest age pine plantations.

Up to 16,000 containerized trees, including non-transgenic trees, could be transferred to this 

field test site and planted in field test plots of up to 20 acres over three years. A field trial of 

the same clone (EH1) was planted at this site on approximately 0.7 acres under BRS permit # 

08-039-102rm. This trial would be transferred to permit # 08-011-106rm to allow 

flowering.

Jasper County Texas Site 2:

This location has been in agriculture for more than 30 years. The test site was used for 

pasture for the past 10 years. Site preparation involved herbicide application to remove 

existing grasses, subsoiling, preparation for irrigation installation, and flat plantings of trees. 

The areas surrounding the test site consist of natural stands of mixed pine and hardwoods.
Page 13 of 98

Up to 8,000 containerized trees, including non-transgenic trees, could be transferred to this 

field test site and planted in field test plots of up to 10 acres over three years. A field trial of 

the same clone (EH1) was established at this site on 1.0 acre under permit 08-039-102rm.

This trial would be transferred to permit # 08-011-106rm to allow flowering.

Jefferson County Texas Site:

This location has been used for managed agricultural production of rice for more than 5 

years. Site preparation will involve herbicide application, subsoiling and planting of trees in 

flat beds. The surrounding areas of the test site consist of rice plantations. Up to 8,000 

containerized trees, including non-transgenic trees, could be transferred to this field test site 

and planted in field test plots of up to 10 acres over three years

Newton County Texas Site 1:

This location has been a managed loblolly pine plantation for at least 30 years. The previous 

plantings were cultivated in beds using standard silvicultural practices and the areas have 

recently been harvested. Site preparation has or will include herbicide application, plowing, 

and planting of trees in raised or flat beds. The areas surrounding the test sites consist of 

managed loblolly pine plantations and/or mixed hardwood stands. Up to 16,000 

containerized trees, including non-transgenic trees, could be transferred to this field test site 

and planted in field test plots of up to 20 acres over three years.

Newton County Texas Site 2:

This location has been a managed loblolly pine plantation for at least 30 years. The previous 

plantings were cultivated in beds using standard silvicultural practices and the areas have 

recently been harvested. Site preparation has or will include herbicide application, plowing, 

and planting of trees in raised or flat beds. The areas surrounding the test sites consist of 

managed loblolly pine plantations and/or mixed hardwood stands. Up to 16,000 

containerized trees, including non-transgenic trees, could be transferred to this field test site 

and planted in field test plots of up to 20 acres over three years.

Newton County Texas Site 3:

This location has been a managed loblolly pine plantation for at least 30 years. The previous 

plantings were cultivated in beds using standard silvicultural practices and the areas have 

recently been harvested. Site preparation has or will include herbicide application, plowing, 

and planting of trees in raised or flat beds. The areas surrounding the test sites consist of 

managed loblolly pine plantations and/or mixed hardwood stands. Up to 16,000 

containerized trees, including non-transgenic trees, could be transferred to this field test site 

and planted in field test plots of up to 20 acres over three years.

Newton County Texas Site 4:

This location has been a managed loblolly pine plantation for at least 30 years. The previous 

plantings were cultivated in beds using standard silvicultural practices and the areas have 

recently been harvested. Site preparation has or will include herbicide application, plowing, 

and planting of trees in raised or flat beds. The areas surrounding the test sites consist of 
Page 14 of 98

managed loblolly pine plantations and/or mixed hardwood stands. Up to 16,000 

containerized trees, including non-transgenic trees, could be transferred to this field test site 

and planted in field test plots of up to 20 acres over three years.

Bay County Florida Site:

This location was used as an intensely managed pasture for more than 15 years, and was 

planted with grasses suitable for cattle grazing. Site preparation involved herbicide 

application to remove existing grasses, subsoiling, preparation for possible irrigation 

installation, and planting of trees in flat beds. The areas surrounding the test site consist of 

experimental forest trees, agricultural crops and less than 25 year-old hardwoods and pine.

Up to 4,000 containerized trees, including non-transgenic trees, could be transferred to this 

site and planted in field test plots of up to 5 acres over the next three years. Field trials of the 

same clone (EH1) were planted at this site on approximately 1.35 acres of test plots under 

permit #08-039-102rm. All tests except approximately 0.2 acre were subsequently 

terminated. The remaining 0.2 acres would be transferred to permit # 08-014-101rm to allow 

flowering.

Columbia County Florida Site:

This location has been a managed pine plantation for more than 20 years. Previous plantings 

were cultivated in beds using standard silvicultural practices. This area of the tract was 

burned in a fire in May 2007. After the fire, the area was raked and bedded by the site owner 

in preparation for re-planting. The test site is within the larger harvested and bedded area, 

which is surrounded by existing pine plantations and additional harvested tracts. Up to 

16,000 containerized trees, including non-transgenic trees, could be transferred to this site 

and planted in field test plots of up to 20 acres over the next three years.

Gadsden County Florida Site 1:

This location has been used as an agricultural research station for more than 10 years for 

conducting research experiments on agricultural crops. Standard silvicultural practices were 

used for site preparation, including herbicide application, plowing and planting of trees in 

raised or flat beds. The areas surrounding the test site consist of mixed pine-hardwood 

forests and pine plantations as well as research plantings of agricultural and horticultural 

crops. There is an experimental plot of non-transgenic Eucalyptus species including E. 

grandis, E. amplifolia, and E. camaldulensis at least 1000 meters away from the transgenic 

test location.

Up to 8,000 containerized trees, including non-transgenic trees, could be transferred to this 

site and planted in field test plots of up to 10 acres over the next three years. A field trial of 

the same clone (EH1) was planted at this site on approximately 0.2 acres under permit # 08-

039-102rm. This trial would be transferred to permit # 08-014-101rm to allow flowering.

Gadsden County Florida Site 2:

This location has been an agricultural research station for more than 10 years. The field has 

been fallow for approximately seven years. Standard silvicultural practices will be used for 
Page 15 of 98

site preparation, including herbicide application, plowing and planting of trees in raised or 

flat beds. The areas surrounding the test site consist of mixed pine-hardwood forests and 

pine plantations, as well as research plantings of agricultural and horticultural crops. Up to 

12,000 containerized trees, including non-transgenic trees, could be transferred to this site 

and planted in field test plots of up to 15 acres over the next three years.

Glades County Florida Site:

Due to changes in experimental plans this site will not be used for field trials of these 

trees and was removed from the permit application per the applicant’s request.

Highlands County Florida Site:

This location was previously used for managed production of citrus for at least 15 years. The 

planting area at this location had been used for field trials of transgenic Eucalyptus for more 

than 6 years. Site preparation involved herbicide application, plowing, and planting of trees 

in flat beds. Areas surrounding the test site consist of less than 5 year-old second-growth 

pine and hardwood with mixed grasses. Field trials of 1.4 acres of some of EH1 translines are 

being grown under issued permit # 08-151-101r under which these trees are allowed to

flower. These trees are now entering their fifth growing season. An additional 2.3 acres of 

tests of different lines with the same EH1 hybrid are being grown under permits 08-039-

102rm (2.0 acres) and 09-070-101rm (0.3 acres).

Up to 8,000 containerized trees, including non-transgenic trees, could be transferred to this 

site and planted in field test plots of up to 10 acres over the next three years.

Marion County Florida Site:

This location has been used as an agricultural research station for more than 5 years for 

conducting research experiments on agricultural crops. Site preparation involved herbicide 

application, plowing, and planting of trees in flat beds. Areas surrounding the test site consist 

of agricultural fields and plantings of horticultural crops. An experimental test of nontransgenic E. amplifolia is planted approximately 200 meters from the test plot location. In 

addition, a field test of 0.3 acres of different lines with the same EH1 hybrid was planted at 

this site under permit 9-070-101rm.

Up to 8,000 containerized trees, including non-transgenic trees, could be transferred to this 

site and planted in field test plots of up to 10 acres over the next three years. A field trial of 

some of the same lines of EH1 clone was established at this site on 0.9 acres under permit 08-

039-102rm. This trial would be transferred to permit # 08-014-101rm to allow flowering. 

Taylor County Florida Site 1:

This location has been a managed pine plantation for over 20 years. The previous plantings 

were cultivated in beds using standard silvicultural practices. The existing pine plantations at 

these sites were harvested by the owner and prepared for re-planting. Additional site 

preparation involved herbicide application, plowing to remove stumps, and planting of trees 
Page 16 of 98

in raised beds. Areas surrounding these test sites consist of managed stands of pine 

plantations.

Up to 16,000 containerized trees, including non-transgenic trees, could be transferred to this 

site and planted in field test plots of up to 20 acres over the next three years. A field trial of 

the same clone (EH1) was planted at this site on approximately 4.3 acres under permit # 08-

039-102rm. This trial would be transferred to permit # 08-014-101rm to allow flowering.

Taylor County Florida Site 2:

This location has been under managed pine plantations for over 20 years. The previous 

plantings were cultivated in beds using standard silvicultural practices. The existing pine 

plantations at these sites were harvested by the owner and prepared for re-planting. 

Additional site preparation involved herbicide application, plowing to remove stumps, and 

planting of trees in raised beds. Areas surrounding these test sites consist of managed stands 

of pine plantations. 

Up to 16,000 containerized trees, including non-transgenic trees, could be transferred to this 

site and planted in field test plots of up to 20 acres over the next three years. A field trial of 

the same clone (EH1) was planted at this site on approximately 3.7 acres under permit # 08-

039-102rm. This trial would be transferred to permit # 08-014-101rm to allow flowering.

Taylor County Florida Site 3:

This location has been under managed pine plantations for over 20 years. The previous 

plantings were cultivated in beds using standard silvicultural practices. The existing pine 

plantations at these sites were harvested by the owner and prepared for re-planting. 

Additional site preparation involved herbicide application, plowing to remove stumps, and 

planting of trees in raised beds. Areas surrounding these test sites consist of managed stands 

of pine plantations. 

Up to 16,000 containerized trees, including non-transgenic trees, could be transferred to this 

site and planted in field test plots of up to 20 acres over the next three years. A field trial of 

the same clone (EH1) was planted at this site on approximately 1.3 acres under permit # 08-

039-102rm. This trial would be transferred to permit # 08-014-101rm to allow flowering.

IV. Alternatives

This EA analyzes the potential environmental consequences of a proposal for APHIS to 

issue two confined environmental release permits to ArborGen to allow the planting, field 

testing and flowering of a GE Eucalyptus hybrid clone engineered to express various 

genes on 28 confined field site locations in the southeastern United States. Specifically, 

these two permits would authorize ArborGen to continue research on GE Eucalyptus

hybrid trees allowed to be planted under APHIS authorized notifications and permits on 

28 sites, to plant additional trees, and to allow all the trees except the trees on one of the 

sites in South Carolina to flower. Therefore 27 of 28 sites originally allowed to be 

planted with the GE Eucalyptus would be allowed to flower. Based upon the permit 
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applications submitted by ArborGen, two alternatives are considered and analyzed in this 

EA: (1) deny the permits and (2) issue the APHIS permits. 

A. No Action – Deny the Permit

Under APHIS–BRS regulations, the Administrator must either grant or deny permits 

properly submitted under 7 CFR part 340. For the purposes of this EA, the No Action 

alternative would be the denial of permit applications 08-011-106rm and 08-014-101rm.

Transgenic Eucalyptus trees (hybrid of Eucalyptus grandis X Eucalyptus urophylla) have 

been previously approved for planting under APHIS authorized notifications, and 

permits. ArborGen has been allowed to plant trees on 28 sites under permit 08-039-

102rm that are requested under the two new permit applications to allow flowering; and 

two field test sites totaling 7.6 acres have been authorized by APHIS to flower under 

permits 06-325-111r and 08-151-101r. All three permits are still active. Under the No 

Action Alternative, the transgenic Eucalyptus plants (hybrid of Eucalyptus grandis X

Eucalyptus urophylla) currently approved by APHIS for confined field release under 

permit 08-039-102rm will continue to not be allowed to flower. The trees could remain 

in the field, but the applicant will be required to either remove developing flowers or 

remove the trees from the field test if removing flowers becomes too difficult. Under this 

alternative, the applicant would not be allowed to gather data on performance of the 

transgenic trees over a multiyear period and the efficacy of the genes in a wide variety of 

environments. (Permits 06-325-111r (as amended) and 08-151-101r , both of which 

already allow flowering, will remain in effect until they expire (unless they are renewed 

under a new permit). These two permits are not affected by the issuance of permits 08-

014-101rm and 08-011-106rm.)

B. Preferred Alternative – Issue the APHIS Permit

The APHIS-preferred alternative is to issue the APHIS confined field release permits for 

a hybrid of Eucalyptus grandis X Eucalyptus urophylla with supplemental permit 

conditions (Appendix V) for the requested three-year period. The permits will need to be 

renewed and subsequently approved by APHIS to allow the transgenic plants to remain in 

the ground beyond the 3 year time period. Under this alternative, APHIS would issue 

two confined environmental release permits to ArborGen to allow the planting, field 

testing and flowering of a GE Eucalyptus hybrid clone engineered to express various 

genes on 28 confined field site locations in the southeastern United States. Specifically, 

these two permits would authorize ArborGen to continue research on GE Eucalyptus

hybrid trees originally planted under APHIS authorized notifications, to plant additional 

trees, and to allow all the trees except the trees on one of the sites in South Carolina to 

flower. Under this alternative, GE Eucalyptus hybrid trees would be allowed to grow and 

flower where the applicant can gather data on performance of the transgenic trees over a 

multiyear period and the efficacy of the genes in a wide variety of environments. This 

alternative would allow the safe development and use of GE organisms under the mission 

of BRS.
Page 18 of 98

V. Environmental Consequences

A. No Action

Under the no action alternative, the applicant would not be allowed to let the trees flower

on 27 sites currently authorized by APHIS for planting under permit 08-039-102rm. At 

this time 15 of the 27 sites have been planted with Eucalyptus hybrid trees. The applicant 

could continue to plant trees under permit 08-039-102rm, but would not allow the trees to 

flower. Preventing flower formation would prove impossible over time because the trees 

grow too tall and too many flowers are produced to remove manually. Therefore the 

trees on these sites under 08-039-102rm would likely be cut down prior to maturity. The 

Eucalyptus trees currently planted under APHIS permits would remain in the ground but 

would likely be cut down within 2 or 3 years to prevent flowering. At each of the field 

sites the locations could remain planted in short rotation Eucalyptus field tests if 

approved by APHIS under a notification or permit, or the sites could be returned to other 

forest tree production or agriculture/forestry research. Some sites could be returned to 

pasture or other agricultural activities. Intense activity including the use of heavy 

machinery for land preparation, general upkeep, irrigation, and fertilization for the 

management of tree plantings and grasses would continue. For forest tree plantings, 

standard silvicultural practices would continue to be used at these sites.

B. Preferred Alternative

Under this alternative, the applicant would be allowed to let GE Eucalyptus trees produce 

flowers and to plant more trees. This means that the trees would remain in the ground for 

at least 3 years and most likely longer since the applicant has indicated that they are 

planning to renew the permits. If the renewal permits are approved by APHIS, the trees 

could be allowed to stay in the ground until maturity or when normally harvested (age 7-

9). The standard silvicultural practices for land preparation, planting, irrigation, and 

harvesting of trees would continue to be routinely used on these sites. 

Potential Environmental Impact of the Preferred Alternative

Alteration in Susceptibility to Disease or Insects – Potential of the Eucalyptus to 

Harbor Plant Pests

Overall impacts on disease or insect susceptibility would be similar to the no action 

alternative. 

As presented in the permit application submitted by ArborGen:

• There has been no intentional genetic change in these plants to affect their 

susceptibility to disease or insect damage. 
Page 19 of 98

• All of the genes have been previously tested in Eucalyptus in existing field tests 

and the permittee has observed no changes in the incidence of pests, beneficial 

insects or pathogens between the transgenic and non-transgenic controls. 

• None of the genes being engineered into the Eucalyptus plants are expected to 

alter the susceptibility of the transgenic Eucalyptus plants to disease or insect 

damage. 

There might be a concern that altered lignin could lead to an increase in insect or disease 

susceptibility, but the results so far with this particular gene do not indicate that this is the 

case (see below). As prescribed in the supplemental permit conditions assigned to these 

permits, periodic monitoring of the field plots will allow the detection of any unexpected 

infestation by plant disease organisms or animal pests. The permittee is required to 

report any such unanticipated effects to APHIS under the terms of the permit - see 7 

CFR 340.4(f)(10)(ii). 

Although the trees originated from New Zealand, the trees were propagated in sterile 

tissue culture and were free of pests upon importation into the U.S. prior to their 

introduction. All materials were handled in accordance with the USDA–APHIS 

requirements for import and quarantine under a USDA–APHIS PPQ Post-entry 

quarantine permit.

Expression of the Gene Products, New Enzymes, or Changes to Plant Metabolism -

Risk of the Gene Products on the Environment

Overall impacts of genes for the selectable marker, cold tolerance, altered flowering and 

altered lignin would be similar to the no action alternative. These same types of genes 

are currently being used in GE Eucalyptus trees being field tested under current APHIS 

notifications or permits for confined field release. The same genes are being expressed 

whether the trees are allowed to flower or not to flower.

Gene used as selectable marker 

The kanamycin resistance selectable marker gene (nptII) engineered into the trees is 

generally accepted as being safe (Fuchs et al. 1993) and has been used in thousands of 

field tests with no evidence that it has led to an increase in plant pest characteristics. This 

gene does not alter the expression of a gene product or change plant metabolism in such a 

way that it would be expected to cause risk to the environment. In a number of instances, 

plants transformed with this gene have been deregulated by APHIS (e.g. corn, petition 

01-137-01p; rapeseed, petition 01-206-02p; cotton, petition 95-045-01p; and papaya, 

petition 96-051-01p). 

Genes conferring cold tolerance

The C-Repeat Binding Factor (CBF) genes are transcription factors that belong to the 

AP2/EREBP family of DNA binding proteins (Riechmann and Meyerowitz 1998) and 

like other transcription factors act as control switches for the coordinated expression of 

other genes in defined metabolic pathways. CBF protein recognizes and binds to a cold-
Page 20 of 98

and drought-responsive DNA regulatory sequence designated as the C-repeat 

(CRT)/dehydration-responsive element (DRE) (Baker et al. 1994), (YamaguchiShinozaki and Shinozaki 1994), which is found in the promoter regions of many coldinducible genes (Maruyama et al. 2004).

A common observation across experiments in which CBF genes are overexpressed in 

transgenic plants is that constitutive expression of CBF negatively impacts a number of 

other traits (Hsieh et al. 2002). In potato, for example, constitutive expression of 

Arabidopsis CBF genes using the CaMV35S promoter was associated with smaller 

leaves, stunted plants, delayed flowering, and reduction or lack of tuber production (Pino 

et al. 2007). In contrast, CBF genes under the control of a cold-induced promoter, rd29A 

(Yamaguchi-Shinozaki and Shinozaki 1993, Kasuga et al. 1999), (Narusaka et al. 2003), 

increased freezing tolerance to the same level as constitutive expression (about 2 °C, or 

~3 °F) while restoring growth and tuber production to the levels similar to wild-type 

plants (Pino et al. 2007). In the rd29A controlled CBF plants the same level of freezing 

tolerance as the CaMV35S versions was observed after only a few hours of exposure to 

low but non-freezing temperatures. These results suggest that using a stress-inducible 

promoter to direct CBF transgene expression could significantly improve freeze tolerance 

without negatively impacting other agronomically important traits. In the case of these 

Eucalyptus trees, the CBF gene is under the control of a cold inducible promoter which 

causes the gene to be expressed under cold temperatures, thus mitigating the potential of 

reduced growth by overexpression. Under this promoter the trees exhibit normal plant 

growth.

The CBF gene is not expected to produce any toxic substances and is not expected to 

alter the characteristics of the engineered plants other than imparting tolerance to cold 

temperatures. These genes do not alter the expression of a gene product or change plant 

metabolism in such a way that it would be expected to cause risk to the environment. 

These genes are in a number of previous and existing field tests and have not produced 

unanticipated phenotypes that would indicate there have been changes to plant 

metabolism leading to increase plant pest characteristics. 

Gene for altered fertility 

The barnase gene has been engineered into other crops that have been previously 

reviewed and addressed in multiple environmental assessments by APHIS. Male sterile 

corn (USDA APHIS petitions for deregulation 95-288-01p, 97-342-01p and 98-349-01p), 

rapeseed (petitions 98-278-01p and 01-206-01p) and chicory (petition 97-148-01p) have 

been reviewed and granted non-regulated status by APHIS. There is no reason to believe 

that the function and expression of this gene will be any different from the plants in 

which it has been previously assessed. There were no toxicity or allergenicity issues 

found with this gene in previous FDA reviews (See BNF Nos. 31, 32, 45, 57 and 66 at: 

http://www.cfsan.fda.gov/~lrd/biocon.html).). The presence of this gene is likely to 

reduce the ability of the trees to produce progeny and thus further reduce the likelihood 

of the release of the regulated article into the environment. In greenhouse tests using 

tobacco and an early flowering model Eucalyptus (E. occidentalis), the applicant has 

found that the barnase gene has demonstrated 100% efficacy in preventing pollen 
Page 21 of 98

formation. In developing flower buds from field grown transgenic Eucalyptus lines 

containing this cassette, 90% of lines showed complete pollen ablation. Recent 

observations from the replicated field study being conducted in Alabama under the 

approved BRS permit (BRS # 06-325-111r-a1) confirm that cold tolerant trees grown at 

the site and allowed to flower did not produce any viable pollen (see also Appendix I). 

Gene for altered lignin

This gene has been engineered into other crops that have been previously released into 

the environment under both notifications and permits. The gene engineered into the 

plants in these field tests has been previously tested in ArborGen field trials for more than 

two years. There might be a concern that altered lignin could lead to an increase in insect 

or disease susceptibility since lignin is often associated with resistance to insects and 

disease organisms (Pederson et al. 2005), but the results of field tests with this particular 

gene have shown no differences in plant pest susceptibility. Growth measurements have 

indicated that trees containing this gene had normal to a moderately reduced growth

phenotype. The trees were also visually inspected on a monthly basis for the presence of 

any insect and disease damage and these observations found that there have been no 

differences in insect or diseases occurrence in the transgenic lines compared to the 

control trees. However, if during the tests there is evidence of increase disease or insect 

susceptibility, the applicant is required to report this to APHIS. The permittee is required 

to report any such unanticipated effects (including excessive mortality or morbidity) to 

APHIS under the terms of the permit - see 7 CFR 340.4(f)(10)(ii).

Non-coding sequences

The transgenic Eucalyptus also contains non-coding regulatory sequences3 derived from 

plants and plant pathogens. The non-coding regions of the plant pathogens will not result 

in the production of an infectious entity or cause plant disease symptoms. None of these 

sequences are expected to pose a plant pest risk.

Method of transformation

The genes were transferred to Eucalyptus via well-characterized laboratory techniques 

that utilize DNA sequences from Agrobacterium tumefaciens to transfer introduced genes 

into the chromosome of the recipient plant (see reviews by (Zambryski 1988, Klee and 

Rogers 1989) A. tumefaciens is a bacterial plant pathogen that can cause crown gall 

disease on a wide range of dicotyledonous plant species. Although some of the DNA 

sequences used in the transformation process were derived from the A. tumefaciens, the 

genes that cause crown gall disease are first removed, and therefore the recipient plant 

does not have crown gall disease. Following transformation, the bacteria are eliminated 

 3 A non-coding sequence is the strand of DNA that does not carry the information necessary to make a 

protein. In this case the non-coding sequences are strands of DNA such as promoters and terminators that 

drive the expression of the gene but do not result in the formation of a protein, which is the product of the 

gene. Therefore promoters and terminators, by themselves, cannot result in the production of a diseasecausing entity.
Page 22 of 98

from the transformed plant tissue, and the DNA sequences introduced into the plant are 

maintained and inherited as any other genes of the plant cell.

Alteration in Weediness characteristics – Potential of the Engineered Eucalyptus to 

be Invasive.

The potential of the engineered Eucalyptus to be weedy and become invasive was 

covered in a previous EA and response to comments for APHIS permit 06-325-111r 

(http://www.aphis.usda.gov/brs/aphisdocs/06_325111r_ea.pdf) and is herein incorporated 

by reference.

The hybrid Eucalyptus EH1 used to produce the transgenic trees has not been shown to 

be weedy or invasive in the U.S (L. Pearson, ArborGen pers. comm.), but no formal 

assessment has been conducted on its weediness or invasiveness potential. None of the 

genes introduced into Eucalyptus code for traits that would be expected to make the GE 

hybrids more weedy or invasive. The genes introduced to affect cold tolerance could 

make the engineered Eucalyptus more adapted to cold temperatures in the southern 

United States, but this trait alone would not impart invasive or weediness characteristics 

(Kolar and Lodge, 2001) to the engineered plants. The trees would be considered weedy 

or invasive if they were to produce many seedlings that were readily spread away from 

the field test sites. Where the non-engineered hybrid Eucalyptus (EH1) has been grown 

in Brazil, on an estimated 400,000 acres planted over 15 years, there has been no 

indication that large numbers of seedlings are being produced and are becoming invasive 

from the commercial plantations (L. Pearson, ArborGen pers. comm. and Luis Silva, 

International Paper Company, Brazil, comment to docket APHIS-2008-0059). 

Eucalyptus generally has difficulty establishing without human intervention, even in 

warmer climates. Eucalyptus is intolerant of shade or weedy competition. In order to 

successfully germinate and establish, Eucalyptus seed need contact with bare mineral soil 

and the removal of competing plants, either as a result of human intervention or naturally 

following a fire event (Meskimen and Francis 1990, Bell and Williams 1997). The areas 

surrounding the field release sites would not be readily conducive to the establishment of 

seedlings because they are managed or unmanaged areas where other plant species are 

growing. So any seeds that attempted to germinate would face competition and would 

likely be unsuccessful. The addition of the cold-tolerance genes are not expected to 

affect the reproductive biology such as seed production or vegetative reproduction 

capabilities. The gene introduced to alter lignin biosynthesis would also not be expected 

to affect seed production or vegetative reproduction capabilities. The selectable marker 

gene, when used previously, did not contribute to weediness or invasive properties of the 

genetically engineered plants. The gene for altered fertility should not contribute to 

weediness or invasive properties and should reduce the ability of the trees to produce 

progeny. None of the traits introduced into the transgenic Eucalyptus will compromise 

the ability to control these plants as weeds.

In addition, the supplemental permit conditions assigned to these permits will limit the 

reproductive capabilities of this GE Eucalyptus outside the confined field trial locations.
Page 23 of 98

Possibility of Gene Flow Within the Field Test

All of the trees in the test plots, including control non-transgenic trees, have the same 

parental genotype EH1. The high level of self incompatibility in Eucalyptus (Campinhos 

et al. 1998, Pound et al. 2002) is expected to significantly reduce the potential for 

crossing4 (gene flow) within the test plots. Seed set from any self pollination is expected 

to be poor, and the vigor of any selfed progeny is also expected to be greatly reduced. In 

experiments conducted in Brazil and Alabama, the control self-pollinated seed obtained 

from this genotype had abnormal morphology and failed to germinate (ArborGen, 

unpublished results). In recent field releases allowed to flower in Alabama and Florida, 

ArborGen has observed a low level of seed production. However, no seedlings have been 

found established beneath the trees or in the surrounding areas (Appendix I). Even if 

seed are produced in the test, several factors in the biology of Eucalyptus would limit the 

potential for seed dissemination. Although Eucalyptus seed is very light and small, it is 

not adapted to wind dispersal and consequently the dispersal of seed is very limited, 

generally being confined within a radius of twice the tree or canopy height 

(approximately 50 meters for a 25 meter tall tree at harvest age)(Cremer 1977, Gill 1997, 

Linacre and Ades 2004). Another consequence of the very small size of Eucalyptus

seeds is that they have very limited reserves and are intolerant of shade or weedy 

competition. In order to successfully germinate and establish, Eucalyptus seed needs 

contact with bare mineral soil and lack of competition either as a result of human 

intervention or naturally following a fire event (Meskimen and Francis 1990, Bell and 

Williams 1997). Eucalyptus plantations are typically established using rooted plantlets 

because of poor establishment using direct seeding methods. Even for the rooted plants, 

competition control is recommended for several months after planting to ensure good 

survival (Meskimen and Francis 1990). Therefore there is limited possibility that 

volunteer seedlings could become established in any unmanaged areas that may be close 

to the site. However, if they were to appear, the supplemental permit conditions for these 

two permits will require that all volunteers be reported to APHIS, found and destroyed to 

prevent any spread of trees from the field release site.

Eucalyptus seeds do not have any dormancy barriers to prevent germination of volunteer 

seeds (Grose 1960, Wellington 1989, Gill 1997) and seed viability and storage of 

Eucalyptus seeds in soil are less than one year (Gill 1997). The Eucalyptus species that 

have become invasive in California are particularly adapted to a Mediterranean climate 

subject to summer fog, which is conducive to seed germination in those species 

(http://ucce.ucdavis.edu/datastore/detailreport.cfm?usernumber=48&surveynumber=182)

. This type of climate does not exist in the Southeastern U.S. In the event that any viable 

seeds are produced and are deposited on in an area conducive to germination from this 

field trial, these seeds would be expected to germinate within 7 – 14 days (Meskimen and 

Francis 1990). In accordance with the supplemental permit conditions for these permits, 

 4 When plants or trees “cross” the male pollen from one tree can pollinate (fertilize) the female ovule (or 

egg) on the same tree or on another tree. Unlike animals, some plants can fertilize themselves when the 

pollen and ovule are produced on the same tree. In this case all the trees are genetically identical (i.e. the 

same clone)(see footnote 1). Eucalyptus has a built-in mechanism that will inhibit self-fertilization. So 

these GE trees are likely to exhibit reduced fertility and reduced numbers of viable seed compared to fully 

sexually compatible Eucalyptus trees since they are genetically identical individuals.
Page 24 of 98

the bordering fields within 100 meters from the edge of the trials will be monitored every 

six months for germinating seedlings by the applicant. This distance is twice the 50 

meter distance that seeds would be expected to be dispersed from a tree at harvest age

(Cremer 1977, Gill 1997, Linacre and Ades 2004). If transgenic seedlings are observed 

they will be destroyed by the applicant either by uprooting or by spraying with EPA 

approved herbicides (e.g., glyphosate or other herbicides to which these trees are 

susceptible) and APHIS will be notified of their occurrence.

Possibility of Gene Flow Outside of the Field Test

Eucalyptus is adapted for insect pollination, with bees being the predominant vector

(Pacheco et al. 1986, Pacheco 1987, House 1997). Under ideal conditions of humidity 

and temperature, viable Eucalyptus pollen can only be found within approximately 100 

meters from the edge of nearest tree stand (Peters et al. 1990, Linacre and Ades 2004). 

Pacheco (1987) verified that bees (Apis spp.) are the most effective pollinators of 

Eucalyptus, with activity increasing up to 100 meters from the beehive, and decreasing 

after this distance. de Assis (1996) indicated that the minimum distance to prevent 

undesirable pollen contamination of seed producing areas is approximately 300 meters. 

Even if bees were to transport pollen farther distances from the field test sites, there are 

no sexually compatible species nearby with which they could cross and produce offspring 

(see description of the field test sites below).

There could be two possible routes of gene flow outside of the confined field test to other 

Eucalyptus species trials. One could be with nearby transgenic Eucalyptus field test trees 

and the other could be with other nearby non-transgenic Eucalyptus species trials.

• Transgenic trials being conducted by the applicant under approved under current 

BRS notifications and permits, of the same hybrid Eucalyptus variety EH1, are 

planted adjacent to or within the proposed field test plot locations at the sites in

Escambia and Baldwin Counties in Alabama; Evans County in Georgia; Saint 

Landry’s Parish, Louisiana; Pearl River County in Mississippi; Jasper County in 

Texas; and Bay, Highlands, Taylor and Gadsden (Site 1) Counties in Florida. The 

applicant has indicated that they are not aware of any commercial plantings of 

compatible Eucalyptus species within 1000 meters of the proposed test plot 

location at any of these sites. Therefore, based upon the limited distance that 

viable pollen is likely to occur outside a tree stand, it is highly unlikely that gene 

flow would occur outside of the confined field test sites at these locations. The 

two field trials in Alabama and Florida that have allowed these hybrids to flower

under an APHIS permit are producing low numbers of viable seeds (see Appendix 

I). Based upon monitoring conducted from the applicant, no volunteer seedlings 

have been observed at these two field trials to date.

• At all test sites in Berkeley, Charleston, and Marlboro Counties in SC; Marshall 

County in MS; Jefferson and Newton Counties in Texas; Columbia and Gadsden 

(except Site 1) Counties in Florida, there are no known existing transgenic or nontransgenic Eucalyptus field trials. The applicant is not aware of any commercial 

plantings of compatible Eucalyptus species within 1000 meters of the proposed 
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test plot location at these sites. Therefore, based upon the limited distance that 

viable pollen (if it were to be produced) is likely to occur outside a tree stand, it is 

highly unlikely that gene flow would occur outside of the confined field test sites 

at these locations.

• At the Bamberg County South Carolina site, there are transgenic trials, covered 

under permit # 08-039-102rm, of the same hybrid Eucalyptus variety EH1 planted 

adjacent to the proposed field test plot location at this site. There are also 

experimental test plots of non-transgenic cold-hardy Eucalyptus species (E. 

macarthurii, E. benthamii, E. viminalis, E. badjensis, and E. dorrigoensis) planted 

by the applicant within 1000 meters of the proposed test plot location. These cold 

hardy Eucalyptus species are highly unlikely to be reproductively compatible with 

the hybrid Eucalyptus variety because of distant phylogenetic relationship and 

asynchronous flowering. E. grandis and E. urophylla, for which hybrids have 

been generated in directed breeding programs, are in the Salignae and Resiniferae 

series, respectively, of Section Transversaria 

(http://plantnet.rbgsyd.nsw.gov.au/cgi-bin/eucclass.pl?gn=Eucalyptus). In 

contrast, E. macarthurii, E. benthamii, E. viminalis, E. badjensis, and E. 

dorrigoensis are far removed genetically from the genotype used in this field trial 

on the evolutionary scale and reside within different Series and in the Section

Maidenaria of genus Eucalyptus (see http://plantnet.rbgsyd.nsw.gov.au/cgibin/eucclass.pl?gn=Eucalyptus for details on sections and series in Eucalyptus). 

The published literature supports the fact that natural hybridization among 

distantly related species within genus Eucalyptus is rare and hybrid inviability 

increases with increasing taxonomic distance between parents (Potts and Dungey 

2004). In accordance with the supplemental permit conditions, the applicant will 

monitor flower development in both the transgenic trial and the non-transgenic 

trial to determine if there is any overlap in the occurrence of mature flowers. If 

overlaps are observed the applicant is required to report their findings to APHIS.

In the unlikely event that viable pollen from the non transgenic Eucalyptus trees 

fertilizes the transgenic trees, any seed dispersal is expected to be within 100 m of 

the test plot. Should any hybridization and viable seed production occur, in 

accordance with supplemental permit conditions, the monitoring for and removal 

of volunteers within 100 m from the edge of transgenic test plot by the applicant 

would effectively eliminate any seedlings that may be produced.

• At the Hardin County Texas site, there are transgenic trials, covered under permit # 

08-039-102rm, of the same hybrid Eucalyptus variety EH1 planted adjacent to the 

proposed field test plot location at this site. Experimental test plots of nontransgenic E. macarthurii, E. benthamii and E. viminalis are planted within 100 

meters of the proposed test plot location. These cold-hardy Eucalyptus species 

are unlikely to be reproductively compatible with the hybrid Eucalyptus variety 

EH1 because of distant phylogenetic relationship and asynchronous flowering.

E. grandis and E. urophylla, for which hybrids have been generated in directed 

breeding programs, are in the Salignae and Resiniferae series, respectively, of 

Section Transversaria (http://plantnet.rbgsyd.nsw.gov.au/cgibin/eucclass.pl?gn=Eucalyptus). In contrast, E. macarthurii, E. benthamii and E. 
Page 26 of 98

viminalis are far removed genetically from the genotype used in this field trial on 

the evolutionary scale and reside within a different Series and in the Section

Maidenaria of genus Eucalyptus (see http://plantnet.rbgsyd.nsw.gov.au/cgibin/eucclass.pl?gn=Eucalyptus for details on sections and series in Eucalyptus). 

The published literature supports the fact that natural hybridization among 

distantly related species within genus Eucalyptus is rare and hybrid inviability 

increases with increasing taxonomic distance between parents (Potts and Dungey 

2004). In accordance with the supplemental permit conditions, the applicant will 

monitor flower development in both the transgenic trial and the non-transgenic 

trial to determine if there is any overlap in the occurrence of mature flowers. If 

overlaps are observed the applicant is required to report their findings to APHIS. 

In the unlikely event that viable pollen from the non transgenic Eucalyptus trees 

fertilizes the transgenic trees, any seed dispersal is expected to be within 100 m of 

the test plot. Should any hybridization and viable seed production occur, in 

accordance with supplemental permit conditions, the monitoring for and removal 

of volunteers within 100 m from the edge of transgenic test plot by the applicant 

would effectively eliminate any seedlings that may be produced.

• At the Marion County Florida site, a transgenic trial, covered under Permit # 08-

039-102rm, of the same hybrid Eucalyptus variety EH1 is planted at the proposed 

field test plot location. An experimental test of non-transgenic E. amplifolia is 

planted approximately 200 meters from the proposed test plot location. This 

Eucalyptus species is unlikely to be reproductively compatible with the hybrid 

Eucalyptus variety EH1 because of its distant phylogenetic relationship and 

asynchronous flowering. Phenology of the Eucalyptus hybrid used in the 

transgenic trial (mature flowers in mid to late summer) and E. amplifolia (mature 

flowers in the spring) indicates that there would not be any overlap in flowering 

times. As noted above, viable pollen dispersal is generally limited to within 100 

m of the tree stand with the minimum distance to prevent undesirable pollen 

contamination of seed producing areas set at approximately 300 m. All trees to be 

planted in this proposed field trial have been shown in previous trials to exhibit 

the reduced fertility trait (see Appendix I). Results from these studies have shown 

that the reduced fertility trait is highly effective in preventing the formation of 

mature viable pollen. Based upon this information, the hybrid Eucalyptus variety 

EH1 are not expected to successfully fertilize the non transgenic E. amplifolia

trees. Observations will be made by the applicant on developing flowers in the 

test site to verify that these trees do have reduced fertility. The applicant will 

monitor flower development in both the transgenic trial and the non-transgenic 

trial to determine if there is any overlap in the occurrence of mature flowers. In 

the unlikely event that viable pollen from the non transgenic E. amplifolia trees 

could fertilize the transgenic trees, any seed dispersal is expected to be within 100 

m of the test plot. Should any hybridization and viable seed production occur, in 

accordance with supplemental permit conditions, the monitoring for and removal 

of volunteers within 100m from the edge of transgenic test plot by the applicant 

would effectively eliminate any seedlings that may be produced.
Page 27 of 98

• At the Gadsden County Florida Site 1, a transgenic trial, covered under Permit # 

08-039-102rm, of the same hybrid Eucalyptus variety EH1 is planted at the 

proposed field test plot location. There are experimental plantings of nontransgenic Eucalyptus species including E. grandis, E. amplifolia, and E. 

camaldulensis which are at least 1000 meters away from the transgenic test 

location. Therefore, based upon the limited distance that viable pollen (if it were 

to be produced) is likely to occur outside a tree stand, it is highly unlikely that 

gene flow would occur outside of the confined field test sites at these locations. 

In addition, even though the hybrid genotype used in these studies for 

transformation may be compatible with E. grandis trees grown in Florida, E. 

grandis produces mature flowers in the fall whereas the hybrid genotype used in 

these tests produces mature flowers in mid to late summer. Therefore, because of 

asynchronous flowering, hybridization of transgenic trees with the E. grandis at 

this site would be virtually negligible.

There are other species of cold-hardy Eucalyptus that can possibly be grown in the 

Southeast U.S. These species include E. neglecta, E. niphophila, E. pauciflora, E. 

camphora, E. nova-anglica, E. macarthurii, E. gunnii and E. cinerea. These could occur 

in the same states as the proposed field trials . Among these species, E. cinerea, also 

known as the silver dollar tree or Argyle Apple, is the most popular species grown for its 

ornamental foliage (http://www.australiaplants.com/Eucalyptus_cinerea.htm). 

The transgenic Eucalyptus hybrids that will be grown in the proposed field trials are not 

likely to be sexually compatible with any of the cold hardy species listed above. For 

example, E. grandis and E. urophylla, for which hybrids have been generated in directed 

breeding programs, are in the Salignae and Resiniferae series, respectively, of section 

Transversaria (http://plantnet.rbgsyd.nsw.gov.au/cgi-bin/eucclass.pl?gn=Eucalyptus). In 

contrast, E. cinerea, and other cold hardy species mentioned above are far removed 

genetically from the genotype used in this field trial on the evolutionary scale and reside 

within different Series and Sections of genus Eucalyptus (see 

http://plantnet.rbgsyd.nsw.gov.au/cgi-bin/eucclass.pl?gn=Eucalyptus for details on 

sections and series in Eucalyptus). Even among the closely related species of Eucalyptus, 

hybridization rates are generally very low (Volker 1995). The published literature 

supports the fact that natural hybridization among distantly related species within genus 

Eucalyptus is rare and hybrid inviability increases with increasing taxonomic distance 

between parents (Potts and Dungey 2004). Where hybridization is possible, it often 

requires significant human intervention in directed breeding/crossing efforts. (Potts and 

Dungey 2004) make reference to the high degree of inviability in F1 hybrids (offspring). 

Inviability of these offspring may be expressed at germination, in the nursery and even 

after planting in the field. Slower germination of hybrid seed often occurs, along with 

reduced survival of germinants in the nursery, and many seedlings have abnormal 

phenotypes. Griffin et al. (1988) surveyed natural and manipulated hybrids in the genus 

Eucalyptus and discussed the challenges of developing even human-made hybrids from 

such wide crosses (in this case E. grandis and E. globulus in sections Transversaria and 

Maidenaria respectively), with only 4.4% of seed germinating and only 3.2% of these 

producing trees that were worthy of further evaluation. To achieve the development of 

viable hybrids sometimes hundreds of hand pollinations must be made to find a viable 
Page 28 of 98

hybrid that will grow normally. An example of the procedures required to make these 

wide-cross hybrids is given in (Barbour and Spencer 2000). 

A further barrier to potential crossing between the transgenic trees with ornamental E. 

cinerea and other species is the expected differences in flowering times between species

(Gore and Potts 1995, Potts et al. 2003). For example, E. cinerea flowers in spring, while 

the transgenic hybrid genotype used in this test initiates flowers in early summer with 

expected maturation in mid to late summer. In the United States, ArborGen data indicate 

that flowering of the clone being tested occurs in the summer.

Based on the above information, there is little if any significant risk for outcrossing to or 

from other Eucalyptus species because: 1) to date the trees that have been allowed to 

flower have shown no mature pollen formation; 2) other species that are or could be 

grown in the area are unlikely to be compatible; 3) it is unlikely that flowering time in 

other species will overlap with the hybrid used in this test and; 4) hybrids, in the event 

that they could form, would be expected to be of very poor vigor.

Possibility of Vegetative Propagation / Persistence Outside of the Field Test

Overall impacts on the possibility of vegetative propagation and persistence outside the 

field test location would be similar to the no action alternative. 

Unlike some other hardwood forest trees, Eucalyptus does not spread in the environment 

via natural abscissions of branches, or cladoptosis. The asexual propagation of shoots via 

rooted cuttings requires specific environmental conditions such as a greenhouse or a high 

humidity environment (Hartney 1980), so it is highly unlikely that any shoots that fall or 

that are removed from the trees would propagate themselves in the wild. 

Suckering (production of shoots from subterranean roots) does not occur in this 

Eucalyptus hybrid. Regrowth of shoots from stumps of felled trees is common and this 

practice, known as coppicing, is used to regrow trees in a plantation after harvest

(Meskimen and Francis 1990). In accordance with supplemental permit conditions this 

regrowth will be managed in this confined field test at termination by devitalizing any 

sprouts that form from the stumps of harvested trees using EPA registered herbicide 

treatments.

Potential of the Eucalyptus in the Field Tests to Become an Invasive Species that 

Threatens Native Plant and Animal Communities.

There could be a concern that adding the cold tolerance trait would make the engineered 

Eucalyptus more adaptive and invasive in the southeastern U.S. It has been hypothesized 

that engineered traits such as cold tolerance could significantly affect the engineered 

variety’s ability to propagate, survive, and impact native ecosystems. 

There is no evidence to date that the untransformed clone of the Eucalyptus hybrid in 

these permits is weedy or invasive in the U.S. (L. Pearson, ArborGen pers. comm.) None 

of the genes introduced into Eucalyptus code for traits that would be expected to make 
Page 29 of 98

the plants more weedy or invasive. The genes introduced to affect cold tolerance could 

make the engineered Eucalyptus more adapted to cold temperatures in the southern 

United States, but this trait in and of itself would not impart invasive or weediness 

characteristics (Kolar and Lodge 2001) to the engineered plants. 

As discussed in previous sections of this EA, there are multiple mechanisms in place that 

would prevent these Eucalyptus hybrids from establishing themselves in the wild. Since 

only one clone is being planted, viable seed set is likely to be limited due to self 

incompatibility. In accordance with supplemental permit conditions monitoring of seed 

set and seedling viability will be required by the applicant during these confined field 

tests so that the extent of seed production and seedling establishment can be determined. 

In addition altered fertility leading to the lack of viable pollen development has been 

engineered into the trees. The addition of the cold-tolerance genes are not expected to 

affect the reproductive biology such as seed production or vegetative reproduction 

capabilities. The gene introduced to alter lignin biosynthesis would also not be expected 

to affect seed production or vegetative reproduction capabilities. The selectable marker 

gene, when used previously, did not contribute to weediness or invasive properties of the 

genetically engineered plants. The gene for altered fertility should not contribute to 

weediness or invasive properties and should reduce the ability of the tree to produce 

progeny. 

In unlikely event that seeds are formed and seedlings are produced, none of the traits 

introduced into the transgenic Eucalyptus will compromise the ability to control these 

plants as weeds so spread of seedlings and trees from the field test sites is highly 

unlikely. Volunteers can be readily identified and controlled. Should any hybridization 

and viable seed production occur, in accordance with supplemental permit conditions, the

monitoring for and removal of volunteers within 100m from the edge of transgenic test 

plot by the applicant would effectively eliminate any seedlings that may be produced. If 

transgenic seedlings are observed they will be destroyed by the applicant either by 

uprooting or by spraying with EPA approved herbicides (e.g., glyphosate or other 

herbicides to which these trees are susceptible) and APHIS will be notified of their 

occurrence.

Impact on Existing Agricultural Practices

Overall impacts on existing agricultural practices would be similar to the no action 

alternative. 

The establishment and growth of these small confined field tests will not have any impact 

on existing agricultural practices because they are solely for research purposes. Current 

agricultural practices will essentially remain unchanged. As indentified by the applicant, 

the field sites that are being proposed under these permits have been used as forest tree 

plantations, pastures, or for forestry and agriculture research and are specifically designed 

for field testing crop plants or forest trees. 

Potential Impacts to Wildlife 
Page 30 of 98

Native floral communities

The field sites in the permit applications are located in Bamberg, Berkeley, Charleston, 

and Marlboro counties, South Carolina; Evans county, Georgia; Baldwin and Escambia 

counties, Alabama; St. Landry's Parish, Louisiana; Marshall and Pearl River counties, 

Mississippi; Hardin Jasper, Jefferson and Newton counties, Texas; and Bay, Columbia, 

Gadsden, Highlands, Marion, and Taylor counties, Florida. These sites are a mixture of 

pasture, crop lands and forested areas. In the unlikely event that viable seeds are formed, 

these areas are unsuitable for the establishment of the Eucalyptus hybrid clone. As 

discussed in previous sections of this EA, Eucalyptus is intolerant of shade or weedy 

competition. In order to successfully germinate and establish, Eucalyptus seed need 

contact with bare mineral soil and the removal of competing plants, either as a result of 

human intervention or naturally following a fire event. With the exception of the field 

test area, the agricultural areas surrounding the field sites are not conducive to the 

establishment of Eucalyptus. The surrounding agricultural and tree crops would provide 

a shady canopy and competition for light and other resources that would impede seedling 

establishment of Eucalyptus. The inhospitable conditions for seed germination, in 

combination with the supplemental permit conditions established for these permits, will 

make the establishment of Eucalyptus in the surrounding area highly unlikely. Should 

any hybridization and viable seed production occur, in accordance with supplemental 

permit conditions, the monitoring for and removal of volunteers within 100m from the 

edge of transgenic test plot by the applicant would effectively eliminate any seedlings 

that may be produced. If transgenic seedlings are observed they will be destroyed by the 

applicant either by uprooting or by spraying with EPA approved herbicides (e.g., 

glyphosate or other herbicides to which these trees are susceptible) and APHIS will be 

notified of their occurrence.

Terrestrial animals

The most likely animals to encounter the transgenic Eucalyptus trees in this confined

field trial would be browsing mammals (e.g., deer), burrowing animals (such as rodents), 

and leaf consuming insects (considered plant pests). In the event of consumption of plant 

material or seeds by other animals, the gene products produced by the selectable marker 

gene and genes of interest do not produce any toxin or have any similarity to known 

toxins (see Section above on - Risk of the Gene products on the Environment). 

Therefore APHIS concludes that the Eucalyptus hybrid would have no adverse impacts 

on any native vertebrate or invertebrate animal species.

Aquatic organisms

Eucalyptus germinates in areas of bare mineral soils and would not be expected to 

establish in aquatic or riparian environments. There is no expectation of toxicological 

effects on any aquatic organism due to the ingestion of the transgenic plant material in 

this confined field trial (see Section above on - Risk of the Gene products on the 

Environment). Therefore, APHIS concludes that the Eucalyptus hybrid would have no 

adverse impacts on any aquatic species.
Page 31 of 98

Potential Impacts by Fire

Overall impacts of fire would be similar to the no action alternative.

Most Eucalyptus communities in Australia have evolved in the presence of periodic fire, 

and fires are an integral part of the Eucalyptus ecosystem (Ashton 1981) (Gill 1997). 

Many Eucalyptus species are known to be highly flammable and depending upon the 

species, location and age, they can be very resistant or susceptible to fire damage (Gill 

1997). Eucalyptus fires can be very hot and move rapidly. The bark catches fire readily, 

and deciduous bark streamers tend to carry fire into the canopy and to disseminate fire 

ahead of the main front (Ashton 1981) (Skolmen and Ledig 1990) (Esser 1993). Other 

features of Eucalyptus that promote fire spread include heavy litter fall, flammable oils in 

the foliage, and open crowns bearing pendulous branches, which encourages maximum 

updraft (Esser 1993, Gill 1997). In the U.S., there have been reports of significant fires 

in California and many have been blamed on the widespread planting of Eucalyptus. 

Fuel buildup occurs very rapidly in unmanaged bluegum Eucalyptus stands in California

which has lead to significant forest fires. The buildup of litter and dead grass are primary 

responsible for the spread of these fires (see Santos: 

http://wwwlibrary.csustan.edu/bsantos/euctoc.htm).). The Forest Service indicates that 

fuel reduction programs and the establishment of firebreaks in Eucalyptus plantings can 

reduce wildfire hazard. (Esser 1993). 

There is a historical risk of forest fire in the southeastern U.S., however, the probability 

that these confined field trials will increase the risk and severity of forest fires in their 

respective locations is very small. These plantings are small (none greater than 20 acres)

and they will be managed by the applicant to prevent litter buildup. These sites are also 

physically isolated from nearby plantations. If they were to catch fire, the fires would 

likely be readily contained. The sites are managed to reduce the risk of fire spreading to 

or from the study areas by maintaining a firebreak between the test plots and adjacent 

forested areas. Depending on local conditions at each site the firebreak may be a road, a 

cultivated strip or a plowed fire line.

Potential Impacts to Human Health

Overall impacts on human health would be similar to the no action alternative.

During the comment period for the EA prepared for permit 06-325-111r, there were 

concerns expressed that Eucalyptus field tests could be a source of Cryptococcus 

neoformans gattii. APHIS conducted a thorough review of C. neoformans gattii and the 

possibility that the field tests could pose a risk to human health (APHIS 2004) EA and 

response to comments for permit 06-325-111r 

(http://www.aphis.usda.gov/brs/aphisdocs/06_325111r_ea.pdf) and concluded that the 

field trial would not lead to a higher incidence of C. gattii in the U.S. and therefore should 

not pose an unnecessary risk to human or animal health.

C. neoformans gattii is a fungal pathogen that is hosted on a variety of species of 

Eucalyptus as well as other tree species (Upton et al. 2007). It causes systemic fungal 
Page 32 of 98

infections in humans, leading to fungal meningitis and death (Datta et al., 2009). C. 

neoformans gattii has been found on a number of Eucalyptus hosts, some of which are 

being grown in commercial plantations and imported and exported for ornamental use. 

People have contracted and died from cryptococcosis in India, Africa, Taiwan, South 

America and California (Datta e al, 2009). C. neoformans infections are found 

particularly in AIDS patients due to their weak immune systems (Chaturvedi et al, 2005),. 

Infections with this fungus are rare in those with fully functioning immune systems. For 

this reason, C. neoformans is sometimes referred to as an opportunistic fungus. There 

was an outbreak of cryptococcal disease on the eastern portion of Vancouver Island, 

British Columbia in 1999 (Datta et al., 2009). The disease was previously only known to 

occur in tropical or semi-tropical climates.

 

It is unlikely that the trees that are the subject of the proposed field release can be a 

source that might introduce the pathogen into the U.S because the trees were derived 

from sterile tissue culture lines. The transgenic Eucalyptus started as a hybrid developed 

in Brazil. In Brazil, small pieces of the tissue derived from the hybrid were put into sterile 

tissue culture and sent to New Zealand for transformation. The transformed lines were 

sent to the U.S as sterile tissue culture lines that were inspected by APHIS Plant 

Protection and Quarantine inspectors prior to entry into the U.S. C. gattii spores readily 

germinate in culture. If C. gattii spores were present in the tissue culture, contamination, 

would be evident and the affected lines would be discarded prior to regeneration of trees 

for introduction into the environment. Another reason it is unlikely that spores could be 

or were ever present in the hybrid lines used in the proposed confined field trial is that in 

the Eucalyptus species where C. gattii is associated, the pathogen is primarily found 

colonizing the bark or decaying wood in hollows of older trees (Kidd et al, 2007). The 

tissue culture used for the Eucalyptus hybrid was not derived from woody tissue nor was 

woody tissue generated during tissue culture. Because the trees were derived from 

tissues that are not known to be a source of the spores and were derived from sterile 

tissue culture lines that by all appearances were free from any fungal contamination, there 

is a negligible risk that the hybrid trees used in the field trial could be or have been 

contaminated with C. gattii. 

The risk that these field trials will result in a higher incidence of the fungus in the U.S. 

and thereby pose a risk to human health is considered to be negligible for the following 

reasons. First, there is not a clear association between E. grandis or E. urophylla and C. 

gattii. Second, there is no reason to believe that the genetic modification of the hybrids 

will alter the association of the trees with C. gattii. Third, the scale of the field tests is 

miniscule compared to the vast expanses of native trees that could potentially harbor the 

pathogen. Based on the above considerations we have concluded that an increase of 

additional acreage planted to Eucalyptus would not impact the likelihood that these field 

trials should lead to a higher incidence of C. gattii in the U.S. and therefore should not 

pose an unnecessary risk to human health. 

Transfer of Genetic Information to Organisms with which it Cannot Interbreed -

Horizontal Gene Transfer to Other Organisms

Overall impacts on horizontal gene transfer would be similar to the no action alternative.
Page 33 of 98

Horizontal gene transfer (HGT) is any process in which an organism incorporates genetic 

material from another organism without being the offspring of that organism. HGT is a 

common phenomenon among bacteria but is not common between higher organisms

(Keese 2008). HGT and expression of DNA from these Eucalyptus hybrid plant species 

to bacteria is unlikely to occur. First, many genomes (or parts thereof) have been 

sequenced from bacteria that are closely associated with plants including Agrobacterium

and Rhizobium (Kaneko et al. 2000, Wood et al. 2001, Kaneko et al. 2002). There is no 

evidence that these organisms contain genes derived from plants. Second, in cases where 

review of sequence data implied that horizontal gene transfer occurred, these events are 

inferred to occur on an evolutionary time scale on the order of millions of years (Koonin 

et al. 2001, Brown 2003). Third, transgene DNA promoters and coding sequences are 

optimized for plant expression, not prokaryotic (i.e., bacterial) expression. Thus even if 

horizontal gene transfer occurred, proteins corresponding to the transgenes are not likely 

to be produced. Fourth, many common transgenes used in plant biotechnology are 

derived from bacteria commonly found in the environment. The FDA has evaluated 

horizontal gene transfer from the use of selectable marker genes and concluded that the 

likelihood of transfer of such genes from plant genomes to microorganisms in the 

gastrointestinal tract of humans or animals, or in the environment, is remote 

http://www.fda.gov/Food/GuidanceComplianceRegulatoryInformation/GuidanceDocume

nts/Biotechnology/ucm096135.htm. Therefore APHIS concludes that horizontal gene 

transfer poses no environmental risk.

C. Cumulative Effects

The field test sites in this permit application have been in agricultural or forest research, 

or in agricultural production or forest tree plantations for 5 to 50 years. Therefore the 

land has been in continuous agricultural or forest tree production at all the proposed field 

sites for at least 5 years prior to these proposed releases and it is reasonably foreseeable 

that if the permit were not issued that the sites would continue to be maintained under 

similar agriculture or forestry production. It is also reasonably foreseeable that the 

applicant may request to further extend the permit for this environmental release for 

additional years beyond the 3 years indicated in the pending permit to observe the growth 

of these trees to maturity. Moreover, APHIS has received a petition for the deregulation 

of these transgenic Eucalyptus trees, however, the environmental effects of that petition 

will be analyzed in a separate NEPA document. The temporary change from agricultural 

crops to a tree crop may result in a temporary change in resident animal and plant 

species, but after harvest and termination of the proposed permits, it is reasonably 

foreseeable that the land will return to agriculture or be replanted to tree production or 

research. At the end of the field test, transgenic plant material will be removed from the 

test site and/or destroyed in accordance with supplemental permit conditions established 

for these permits. Therefore the only past, present, and reasonably foreseeable actions 

associated with the locations for the proposed releases under permit are those related to 

agricultural or forest tree production. APHIS has determined that there are no past, 

present, or reasonably foreseeable actions that would aggregate with effects of the 

proposed action to create cumulative impacts or reduce the long-term productivity or 

sustainability of any of the resources (soil, water, ecosystem quality, biodiversity, etc.) 
Page 34 of 98

associated with the release sites or the ecosystem in which they are situated. No 

resources will be significantly impacted due to cumulative impacts resulting from the 

proposed action. 

D. The Degree to Which the Possible Effects are Highly Uncertain or 

Involve Unique or Unknown Risks

Potential Effects of Growing Eucalyptus on Soil Hydrology

Eucalyptus is recognized as having impacts on hydrology and large widespread plantings 

could have potential impacts on hydrology in the southeastern United States (Farley et al. 

2005a). Since large plantings of Eucalyptus have not been grown in many parts of the 

southeastern US (other than southern Florida) the potential impacts of such plantings on 

hydrology are unknown. APHIS requested additional information on potential impacts of 

hydrology from ArborGen and also consulted with the USDA Forest Service to assess the 

potential impacts of planting Eucalyptus on hydrology. The additional information 

supplied by ArborGen and the Forest Service are included as Appendices II and III. The 

document supplied by the Forest Service represents only their opinion on the potential 

impacts of these field tests on hydrology and does not represent the position of the USDA 

on the pros and cons of deploying Eucalyptus as a biofuel, bioenergy or fiber crop.

The Forest Service indicates that planting large-scale Eucalyptus plantations may 

potentially lower the water table, and affect groundwater recharge and stream flow 

dynamics. Eucalyptus is very efficient at using water. It can produce more biomass per 

unit water consumed than native southeastern pines; however, their extremely rapid 

biomass production has proportionally higher transpirational costs and therefore greater 

water use. The Forest Service has estimated that a mature Eucalyptus plantation growing 

in southwest Georgia could potentially transpire 882 mm per year, exceeding all other 

forest types on average by a factor of 2.5. Eucalyptus transpiration could exceed that of 

pine plantations by a factor of 1.6, and previous pasture land by a factor of 3.5. The 

comparison with agricultural crops is more variable where Eucalyptus transpiration may 

be greater or lesser than that of crop plants depending on the crop, the growing season, 

and the management practices.

Eucalyptus has a dimorphic rooting pattern which means that it has surface roots that 

draw water from the surface as well as deep roots which draw water from deep within the 

soil. The mean maximum rooting depth for Eucalyptus is 15 meters, which is a 

characteristic of a dimorphic rooting pattern. In contrast, mean maximum rooting depths 

of pine plantation (P. taeda and P. elliottii) and grass species are 3 meters and 2.6 meters, 

respectively (Canadell et al. 1996a). According to the Forest Service, conversion to

Eucalyptus on sites where the water tables are less than 10 meters will likely lower downslope water tables via direct means (i.e., direct use of ground water by deep roots), affect 

groundwater-aquifer dynamics, and result in evapotranspiration rates that exceed 

precipitation input, as have been reported for this species in other locations (Calder et al. 

1997a).
Page 35 of 98

Recent research suggests that Eucalyptus plantations would reduce stream flow more 

than pine plantations, and could potentially eliminate low flows. In a review of more 

than 20 catchment5 conversion studies, Farley and others (Farley et al. 2005a) showed 

that converting existing vegetation to Eucalyptus plantations reduced stream flow by 20% 

more than converting it to a pine plantation. This review also showed that the loss of low 

flows were more complete for Eucalyptus plantations compared to pine plantations 

(100% vs. ~80% reduction of low flows). Elimination of low stream flows could have 

important ramifications for threatened and endangered aquatic species, such as the gulf 

strain striped bass, and species of endemic freshwater mussels (Golladay et al. 2004a, 

Couch and McDowell 2006b). 

Due to a lack of available data in the southeastern U.S. on planting Eucalyptus, it is 

difficult to determine the significance of the effects on hydrology if large acreage of 

Eucalyptus were to be planted. The Forest Service has indicated that collection of data 

and modeling will be useful to determine the long-term impacts of planting large 

acreages of the genus. The Forest Service has also pointed out that the significance of the 

impact on groundwater and stream flow will depend greatly on the area extent, size, and 

spatial distribution of the plantations. For example, a few small (less than 10 hectares, 

i.e. approximately 25 acres) and well-dispersed plantations may only have very localized 

impacts and negligible impacts at the watershed scale.

The field test sites requested under the two permits are well dispersed and are limited in 

size (none are greater than 20 acres) and it is anticipated that they are not likely to have 

significant impacts on hydrology. At the request of APHIS, ArborGen has supplied data 

indicating the maximum size of each of the plantings at each site, the individual 

watersheds where the plantings occur, the area of the watershed, how much of the 

watershed will be occupied by the field tests, the location of the closest primary and 

secondary streams, and the location of any critical habitat for Federally listed threatened 

and endangered species within the watershed. 

Using the 8 digit HUC (Hydrologic Unit Code) as the Watershed to be analyzed, the data 

provided by ArborGen show that none of the sites occupy more than 0.03% of any given 

watershed. The closest critical habitat for an aquatic species (such as a fish and mussel) 

is 6 kilometers at one location and ranges from 6 to 90 km for any of the sites having any 

proximity to habitats that could be impacted. There are no nearby threatened or 

endangered plant species that could be impacted by hydrological effects (see also 

Appendix IV). Any effects would be very localized on existing nearby agricultural and 

forestry plantings. Therefore APHIS concludes that while the effects on hydrology, 

including the watershed and aquifers, are unknown and uncertain for very large plantings 

of Eucalyptus, these small-scale field tests are unlikely to have any significant negative 

impacts on hydrology and on native flora and fauna.

Potential Allelopathic Effects of Eucalyptus

 5 A catchment or drainage basin is an extent of land where water from rain or snow-melt drains downhill 

into a body of water, such as a river, lake, reservoir, estuary, wetland, sea or ocean. The drainage basin 

includes both the streams and rivers that convey the water as well as the land surfaces from which water 

drains into those channels, and is separated from adjacent basins by a drainage divide.
Page 36 of 98

Allelopathy refers to “any process involving secondary metabolites produced by plants, 

microorganisms, viruses and fungi that influence the growth and development of 

agricultural and biological systems” (See: International Allelopathy Foundation -

http://www.allelopathy-journal.com/allelopathy.aspx). Allelochemicals from plants are 

released into the environment by exudation from roots, leaching from stems and leaves, 

or decomposition of plant material. Allelopathy can have both negative and positive 

impacts on the environment (Eljarrat and Barceló 2001, Xuan et al. 2005, Kohli et al. 

2006). There has been increased research activity in this area, one of which is taking 

advantage of plants that produce allelopathic compounds in developing agroforestry and 

sustainable agriculture systems (Kohli et al. 2006, Narwal 2006). Allelopathy has been 

demonstrated in many commercially important tree species including Acacia, Ailanthus 

Eucalyptus, Juglans, Quercus, Leucaena, Pinus, Picea, Aibes, Populus and Acer; and has 

been demonstrated in agronomic crops such as rye, wheat and alfalfa (Nandal et al. 1994, 

Ferguson and Rathinasabapathi 2003, Reigosa and Gonzáles 2006, Mallik 2008).

There have been extensive studies conducted on allelopathy in Eucalyptus and there are 

several comprehensive reports and review articles on this genus (Ong 1993, Sunder 1995)

(Nandal et al. 1994, Davidson 1995, White 1995). Eucalyptus species are known to 

produce chemical compounds that are required by the plant for defense against herbivores 

and pathogens. There are several studies in the literature that demonstrate the negative, 

positive and neutral allelopathic interaction of Eucalyptus species and their hybrids with 

other crop plants (Sanginga and Swift 1992) (Khan et al. 2004) (Espinosa-Garcia et al. 

2008). These interactions vary greatly depending upon the crop species and conditions 

under which they are grown. There is inconclusive data as to whether these compounds 

produced by Eucalyptus are exclusively responsible for allelopathic influence on 

understory vegetation in Eucalyptus plantations. Most allelopathic studies in Eucalyptus 

species have involved laboratory experiments with extracts obtained from different plant 

parts or leaf litter to investigate allelopathic effects on seed germination and growth in 

potted plants. These laboratory bioassays and pot culture studies may or may not be 

applicable to field conditions. The perceived allelopathic effects observed in the field on 

growth of understory or adjacent intercropped food crops could also result from 

competition for water, nutrients and light.

Allelopathy tends to be an inexact science and many studies in allelopathy are 

inconclusive and difficult to interpret due to potential interactions with other aspects of 

the environment. For example in a recent study, (Nandal and Dhillon 2005) tested the 

allelopathic effects of poplar (Populus deltoides) leaf extracts on germination and growth 

of ten wheat varieties under laboratory conditions. They reported that lower concentration 

of leaf extracts from poplar had stimulatory effects on root length in all wheat varieties 

whereas higher concentrations adversely affected germination and seedling growth of 

some of the wheat varieties tested. In a field experiment, the performance of all ten 

wheat varieties was also evaluated under four different poplar spacings in an agrisilviculture system. Although the grain yield of wheat varieties was significantly lower 

under all spacings of poplar compared to controls, yields increased significantly with 

increased spacing of poplar, possibly due to reduced competition for light and nutrients. 
Page 37 of 98

However, no correlation was found between the laboratory bioassay using leaf extracts 

and the field studies.

In a recent study, the allelopathic interaction of Eucalyptus grandis, E. urophylla and E. 

grandis x urophylla on the germination and early growth of four annual crops (maize, 

bean, watermelon and squash) was investigated (Espinosa-Garcia et al. 2008). Soil 

samples were collected from different soil horizons and at varying distances from 

Eucalyptus trees growing at the plantation edge and used for growth studies in pots. The 

dried soil samples used for growth studies were also analyzed for total soluble phenolics 

present in the soil. The study showed that soil samples from different plantations had 

differential effects ranging from no effect, to slightly inhibitory, to a stimulatory effect on 

germination and radicle6 growth of test crops. Among the three Eucalyptus species tested, 

the soil samples from E. grandis x urophylla plantations had an inhibitory effect on 

germination of maize, bean and watermelon but had a stimulatory effect on squash. The

soil from E. grandis plantations had an inhibitory effect on squash. The total soluble 

phenolics varied in different soil samples but did not explain the differential effects on 

the test crops. The authors concluded that soil samples collected from plantations of 

Eucalyptus species contained allelochemicals that affected germination and early growth 

of some annual crops but such effects could be avoided by planting crops at a distance of 

15 meters away from the edge of plantations.

Even though the Eucalyptus under this permit could demonstrate allelopathic properties, 

the presence of any allelochemicals is not going to make the Eucalyptus planted under 

these permits more invasive or present a plant pest risk. Since all these field tests are 

confined and limited in size, any allelopathic effects should be small. As a standard 

silvicultural practice, herbicides will also be used within the field test sites and any of 

their effects on understory vegetation will be as severe or more severe than any 

allelopathic effects. In the future, should any negative allelopathic or other competitive 

interactions be observed under field conditions outside of the immediate field tests sites, 

these could be mitigated by adjusting the tree spacing, irrigation and fertilization 

practices or by planting the field tests at least 15 meters away from any agronomic crops 

or sensitive areas. Any unusual observations at the field test sites are to be reported to 

APHIS under the supplemental permit conditions of the permit; including any indications 

of allelopathic effects.

E. Risks to Threatened and Endangered Species

APHIS analyzed the potential for effects from the preferred alternative of this EA on 

federally listed threatened and endangered species (TES) and species proposed for listing, 

as well as designated critical habitat and habitat proposed for designation, as required 

under Section 7 of the Endangered Species Act. APHIS worked with the United States 

Fish and Wildlife Service to obtain species lists and information and critical habitat 

information for the proposed field site locations. After analyzing the potential for any 

effect, APHIS has reached a determination that the proposed environmental release will 

 6 The radicle is the first part of a seedling (a growing plant embryo) to emerge from the seed during the 

process of germination. It is an embryonic root.
Page 38 of 98

have no effect on federally listed threatened or endangered species or species proposed 

for listing, and no effect on designated critical habitat or habitat proposed for designation. 

Consequently, consultation with the United States Fish and Wildlife Service is not 

required for the action described in the preferred alternative of this EA. Appendix IV

includes the BRS analysis of threatened and endangered species in the areas of the field 

releases.

F. Other Considerations

Consideration of Executive Orders, Standards and Treaties Relating to 

Environmental Impacts.

Executive Order (EO) 12898, "Federal Actions to Address Environmental Justice in 

Minority Populations and Low-Income Populations," requires Federal agencies to 

conduct their programs, policies, and activities that substantially affect human health or 

the environment in a manner so as not to exclude persons and populations from 

participation in or benefiting from such programs. It also enforces existing statutes to 

prevent minority and low-income communities from being subjected to 

disproportionately high and adverse human health or environmental effects.

EO 13045, “Protection of Children from Environmental Health Risks and Safety Risks,” 

acknowledges that children may suffer disproportionately from environmental health and 

safety risks because of their developmental stage, greater metabolic activity levels, and 

behavior patterns, as compared to adults. The EO (to the extent permitted by law and 

consistent with the agency’s mission) requires each Federal agency to identify, assess, 

and address environmental health risks and safety risks that may disproportionately affect 

children. 

Each alternative was analyzed with respect to the above EO 12898 and 13045. The 

human health and environmental impacts of the action alternatives are presented in 

Section V of this EA. No human health or environmental effects were identified for any 

of the action alternatives that would have a disproportionate adverse effect or that would 

exclude a particular group of persons or populations, including minority and low-income 

populations, or children, from expected benefits.

EO 13112, “Invasive Species”, states that federal agencies take action to prevent the 

introduction of invasive species and provide for their control and to minimize the 

economic, ecological, and human health impacts that invasive species cause. As 

presented in Section V of this EA, the hybrid species of Eucalyptus being grown is not 

considered an invasive species and does not establish itself without human intervention. 

Based on historical experience with the Eucalyptus in these field tests, the engineered 

plant is not expected to have an increased invasive potential. 

Executive Order 12114, “Environmental Effects Abroad of Major Federal Actions”

requires Federal officials to take into consideration any potential environmental effects 

outside the U.S., its territories and possessions that result from actions being taken. 

APHIS has given this due consideration and does not expect an environmental impact 
Page 39 of 98

outside the United States should APHIS choose any of the two alternatives. These 

confined field tests are being conducted in the continental U.S. and would not be 

expected on have environmental effects outside of the U.S.

Migratory Bird Treaty Act, 1918 as amended and Executive Order 13186. Migratory 

birds include all native wild birds found in the United States except the house sparrow, 

starling, feral pigeon, and resident game birds such as pheasant, grouse, quail, and wild 

turkeys. A reference list of migratory game birds is found in Title 50, Code of Federal 

Regulations, Part 10. The Migratory Bird Treaty Act makes it unlawful for anyone to 

kill, capture, collect, possess, buy, sell, trade, ship, import, or export any migratory bird, 

including feathers, parts, nests, or eggs. Executive Order 13186 “Responsibilities of 

Federal Agencies to Protect Migratory Birds” requires Federal officials to consider the 

impacts of planned actions on migratory bird populations and habitats for all planning 

activities. APHIS has determined that it is reasonable to assume that the activities at the 

field test sites such as planting, collecting samples and eventual harvest of the trees 

should have no adverse impact on migratory birds since they would not be expected to

nest or permanently inhabit these types of field test sites.

Consistency of Proposal with other Environmental Requirements:

The proposal is believed to be consistent with other environmental requirements. This 

environmental assessment was prepared in accordance with: (1) The National 

Environmental Policy Act of 1969 (NEPA), as amended (42 U.S.C 4321 et seq.); (2) 

regulations of the Council on Environmental Quality for implementing the procedural 

provisions of NEPA (40 CFR parts 1500-1508); (3) USDA regulations implementing 

NEPA (7 CFR part 1b); and (4) APHIS’ NEPA Implementing Procedures (7 CFR part 

372).

G. Conclusion

As outlined under the Purpose and Need sections of this document, this EA was prepared 

because it was necessary for APHIS to evaluate the potential environmental impacts 

resulting from the increased number of locations and size of the releases of flowering 

Eucalyptus, which could potentially lead to a lack of confinement of the field tests and

other impacts to the environment. APHIS has evaluated the permit applications to 

determine whether the environmental release, with appropriate conditions imposed, can 

be carried out while preventing the dissemination and establishment of plant pests. After 

preparing this draft EA, APHIS has concluded that even though there is an increase in the 

number of sites where trees will be allowed to reach maturity and flower, over those 

already allowed to flower under permits 06-325-111r and 08-151-101r, there is no 

substantially greater risk of loss of confinement and risk to the environment. APHIS 

concludes that the releases will remain as confined field tests and that the genetically 

engineered trees will not pose a significant plant pest risk. In addition, APHIS concludes 

that granting permits will not significantly affect the quality of the human environment. 

No threatened and endangered species or critical habitat should be impacted by letting the 

trees reach maturity and flower at the increased number of locations.
Page 40 of 98

VI. Listing of Agencies and Persons Consulted

James M. Vose - USDA-Forest Service Coweeta Hydrologic Laboratory, Otto, NC

Chelcy R. Ford - USDA-Forest Service Coweeta Hydrologic Laboratory, Otto, NC

Jody Smithen – US Fish and Wildlife Service – Daphne, Alabama Field Office

Kathy Chapman - US Fish and Wildlife Service - Coastal Georgia Field Office

James Harris – US Fish and Wildlife Service – Lacombe, Louisiana Field Office

Laura Zimmerman - US Fish and Wildlife Service – Charleston, South Carolina Field 

Office

Caroline Stahaller - US Fish and Wildlife Service - Panama City, Florida Field Office

Brad Rick - US Fish and Wildlife Service - Vero Beach, Florida Field Office

Candice Martino - US Fish and Wildlife Service - Jacksonville, Florida Field Office

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APPENDIX I: Status of Existing Field Tests Allowed to Flower

The applicant has been allowed to let transgenic Eucalyptus trees flower under APHIS 

permits 06-325-111r-a2 and 08-151-101r. These trees have the same genetic constructs 

as those Eucalyptus hybrid trees under APHIS consideration in the two permit 

applications submitted by ArborGen. The following is a summary of information that has 

been collected by the applicant and provided to APHIS as part of their annual reports 

under the existing permits that allow flowering.

All transgenic and non-transgenic trees in these field tests, spanning 6.2 acres in Alabama 

and 1.4 acres in Florida, produced mature flowers in late August-early September of 2007 

and/or 2008. On the 25-30 foot tall trees in these tests there were estimated to be several 

thousand flowers on each tree. No differences were noted in flower formation in 

transgenic trees compared to non-transgenic controls (the same parental genotype). 

In January 2008, mature (but not yet opened) capsules were collected from nontransgenic and transgenic trees under permit 06-325-111r. Replicate samples were 

collected from transgenic trees and non-transgenic controls in two separate blocks. 

Capsules from a subset of transgenic trees plus non-transgenic controls were dried in the 

laboratory and allowed to open to evaluate the presence of seed or seed like structures in 

the capsules. Approximately 100 capsules for each of the two replicate samples were 

analyzed. Microscopic examination of the material inside the capsules did not show any 

seed or seed like structures in capsules of either non-transgenic or transgenic lines. 

Controlled germination studies of the material extracted from the capsules did not 

produce any germinating seeds. Observations from the replicated field study being 

conducted in Alabama under permit No. 06-325-111r confirm that cold tolerant lines 

grown in this field test also did not produce any pollen. The results to date have shown 

that the barnase gene that has been engineered into these trees is effective at preventing 

pollen formation.

After the initial draft EA was published, the applicant submitted annual reports for 

permits (06-325-111r-a2 and 08-151-101r) as required under the supplemental 

conditions. The transgenic and non-transgenic trees planted under these permits in 

Baldwin County, Alabama and Highlands County, Florida were previously allowed to 

flower. The following is a summary of this report.

In Alabama, the transgenic trees produced several hundred to a few thousand flowers per 

tree whereas the non-transgenic controls had much fewer flowers as a result of significant 

cold damage. In Florida, there was no difference in the number of flowers produced by 

both transgenic and non-transgenic trees. Mature seed capsules, prior to opening, were 

sampled from select trees in three field tests in Baldwin County, Alabama and trees from 

the single field test in Highlands County, Florida in early March 2009. Samples 

consisted of approximately 70 to 100 capsules collected from different positions in the 

crown. The capsules were returned to ArborGen’s greenhouse facility where they were 

dried and contents of the capsules were extracted and stored at 4°C. A controlled 

germination test was conducted using approximately 0.1 g of the extracted contents of 
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each sample spread on moist filter paper in a standard Petri dish. Open pollinated seed of 

EH1 obtained from Brazil were germinated as control seedlings for comparison.

Of the samples collected and analyzed from trees in Baldwin County, AL, approximately 

4% of the seeds showed a low level of germination while approximately 83% of the 

samples collected from trees in the field trial in Highlands County, FL, showed 

germination, including samples from both transgenic and non-transgenic control trees. 

For both transgenic and non-transgenic tree samples analyzed, the applicant observed 2-8 

seedlings for each 0.1g sample plated for germination. In the literature, for the same 

sample size tested for E. grandis 31 to 65 seedlings have been reported. The applicant 

indicates that, as expected from limited self pollination that may have occurred in these 

trees, the number of seedlings observed in these tests is much lower than would be 

expected for open pollinated seeds. 

The controlled germination tests from samples collected in spring 2009 from field trials 

at two sites indicate that a very low level of seedlings can be produced from these trees. 

The number of seedlings produced is significantly less than what would be expected for 

an open-pollinating mixed stand of Eucalyptus. So far the data are consistent with the

hypothesis that limited self pollination can occur from viable pollen produced by the nontransformed control trees. The applicant has also not observed any volunteer seedlings in 

or around the test sites. Volunteer monitoring and reporting the presence of volunteers is 

required in the supplemental permit conditions.

Monthly field test monitoring observations have not identified any differences in diseases 

and insects or other non-target organisms between the transgenic and non transgenic trees 

in the field test.
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APPENDIX II: Hydrology considerations for planted 

Eucalyptus submitted by ArborGen LLC

Submitted by ArborGen. LLC to USDA APHIS BRS in support of consideration for

approval of permits for field trials of Eucalyptus at multiple sites.

August 12, 2008

Introduction

The relevant scientific literature and conclusions drawn by experts in the field of

hydrology, ecology, and plantation management on the hydrology of Eucalyptus

plantations are discussed in this document. Extensive research on hydrology has been

conducted in countries where large plantations of Eucalyptus have been established for

many years, including India, China, South Africa and Brazil, as well as in its native

Australia. The FAO, in response to the criticisms and concerns expressed about

Eucalyptus plantations, has developed several expert reviews on the ecological impacts of

Eucalyptus (discussed below).

The main hydrological concerns voiced against Eucalyptus plantations are that they

deplete water supplies. The authors of an early FAO report (Poore and Fries, 1985) noted

that these same criticisms would apply equally to any other plantation tree species, and

that society tends to judge more harshly forestry crops relative to agricultural crops. As

stated in Poore and Fries, 1985, “... most crops in many parts of the world are of foreign

origin (wheat, maize, rice, potatoes, manioc, rubber, oil palm, coconut and many others.

No one is surprised either that the soil under agricultural crops becomes depleted if these

are continuously cropped without adding fertiliser. But both of these features are

considered grounds for criticism in forestry.” Ironically, Eucalyptus evolved to be water

efficient as the Australian continent itself became more dry (Davidson, 1995). In fact

Eucalyptus uses less water per unit weight of biomass produced than do other kinds of

trees (Chaturvedi, 1987) and many agricultural crops (Davidson, 1995).

It is also important to take into account the breadth of the genus Eucalyptus, often

referred to broadly as eucalypts, where different species have different characteristics

which may prove detrimental or beneficial under different situations. There are several

different ecological situations globally in which eucalypts may be planted: in place of

existing closed forest; in place of other natural vegetation such as savannah, scrub or

grassland; on degraded or waste land either as a potential crop or to assist in the control

of erosion or salinity; within agricultural land as shelter belts or as components of

agroforestry systems, or as intensively managed crops for wood production. It is

important to understand the particular application in order to evaluate the potential

ecological impact of Eucalyptus in a scientific manner. For example, the effect on soil

moisture content and the water table can only satisfactorily be judged with reference to

the pre-existing conditions before the establishment of the planting of Eucalyptus. In the
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case of field trials of Eucalyptus requested in ArborGen’s permit applications, all sites are

on land previously managed for forestry, agricultural production or maintained as pasture

land.

The Water Cycle

In considering the potential hydrological impacts of Eucalyptus it is important to put

these in the context of the water cycle. Poore and Fries (1985) provide a good overview

of the water cycle (see Figure 1 in Poore and Fries, 1985, or see Figure 1-1 in NRC,

2008). One key variable is the amount of rain that is intercepted by the canopy and is

then evaporated back into the atmosphere. As a result, such intercepted water does not

contribute to water in the soil. Of the water that does reach the soil some of this is

absorbed while some fraction runs off the surface or is evaporated. A certain amount of

water is maintained in the soil layer against the forces of gravity (called the 

‘fieldcapacity’ and dependant on soil texture and organic content) while any excess 

drains to the water table, the level at which the soil is permanently saturated. Depending 

on the depth of the water table it may be accessed by deep rooting plants such as trees. 

Even without roots that reach to the water table, plants may be able to access water from

deeper, wetter soil layers through capillary action, depending on the soil type, where

there can actually be an upward movement of water.

Through normal transpiration plant roots take up available water which is transported

through the stem to the leaves, the majority of which is lost to the atmosphere.

Evapotranspiration is the total water returned to the atmosphere through transpiration and

evaporation from the ground, bodies of water, plus intercepted water in the canopy. The

relative rates of evapotranspiration and precipitation are often compared in assessments

of hydrological systems. Where precipitation exceeds evapotranspiration then there is a

net water gain to groundwater or downstream systems. Where evapotranspiration

exceeds precipitation then the available water resources may be depleted. It is important

to understand, as pointed out in Poore and Fries (1985), that water loss is a “price that

plants must pay for growth”. When stomata in plant leaves close then photosynthesis and

growth both cease. In general terms, the rate of growth or biomass production in a plant

is proportional to its water use. Consequently fast growing trees, of any species, use

large quantities of water. As described below, Eucalyptus is actually more efficient in

terms of water used per gram of biomass produced than many other tree species.

Hydrology of Eucalyptus – Key Literature Reviews

There has been extensive literature published on the hydrology of Eucalyptus. Google

Scholar for example lists over 60,000 hits for the keywords ‘Eucalyptus’ and ‘water’. It

is therefore not possible to provide summaries of the entire breadth of the literature.

Several review articles are available including reviews sponsored by FAO that assessed

the ecological impacts of Eucalyptus plantations, including analysis of the impact of

Eucalyptus on hydrology. We provide here a summary of these reviews together with

data from some specific reports (see below) where the hydrology of Eucalyptus has been

studied in detail. FAO has also released two annotated bibliographies (FAO, 2002a,

2002b) that collate and summarize publications on environmental, social and economic
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impacts of Eucalypts, and which include many references to water use. While there are

specific examples and geographic regions where Eucalyptus (and other trees) can

negatively impact hydrology, in general, the literature indicates that Eucalyptus can be

grown in a sustainable manner and that its associated water use is not a major ecological

threat. Most of the reviews indicate that soil and water characteristics of the site should

be taken into consideration when establishing and maintaining a Eucalyptus plantation, in

the same way that would be appropriate for plantings of agricultural crops.

The first FAO review was published in 1985 by Poore and Fries. At that time the authors

suggested that there were relatively few existing studies in several important areas

including hydrology (Lima, 1984; Poore and Fries, 1985). Where comparative studies

showed that for dry alpine conditions the water regime for Eucalyptus did not differ from

adjacent grasslands (see Lima, 1984), this was attributed to Eucalyptus’ ability to control

the rate of transpiration, an evolutionary adaptation for survival of drought stress which is

often typical of the rainfall regimens of their native habitats. For deep soils and higher

rainfall Eucalyptus plantations might be expected to reduce streamflow or groundwater

recharge but that this is comparable to these same effects in pine plantations. In contrast,

the water intercepted and re-evaporated by the foliage, and therefore not available to the

soil, is less for Eucalyptus when compared to pines, due in part to the near vertical

orientation of leaves in Eucalyptus (Whitehead and Beadle, 2004). It was concluded that

the conditions of a particular site need to be taken into account as well as balancing local

demands for forest products and water.

By 1993, at an FAO sponsored regional Expert Consultation on Eucalyptus (White et al,

1995) more information was available about hydrology and Eucalyptus plantations.

These experts presented their experiences with Eucalyptus plantations from Asia. The

report recognized the potential benefits of Eucalyptus and noted that many of the

criticisms of the species were based on inappropriate government policies on

afforestation or social concerns rather than the biology of the trees themselves (see also

Casson, 1997). Calder et al (2004) highlights that many early policies were based on

public misconceptions about the impacts of forests on water. With regard to hydrological

effects on intercropping with other species, the experts in the FAO report concluded that

while Eucalyptus can have negative effects in drier climates, in regions where rainfall is

above 1,200 mm/year this is not expected to be a problem. The report suggests that for

Eucalyptus plantings in those regions where water is scarce or demanded by other

sectors, biomass production could be adjusted to match the amount of water available, for

example by planting fewer trees per unit area or by thinning existing plantations.

As part of this expert consultation Sunder (1995) reported that the overall use of water by

Eucalyptus is limited to the total rainfall of the area, in the absence of access of the tree to

the water table. He concluded that there is an equilibrium between rainfall and

evapotranspiration in Eucalyptus and that this does not differ significantly from other

trees. As an example, monthly evapotranspiration of an E. globulus plantation in

Portugal was the same as that of a natural open stand of cork oak (Quercus suber) with a

developing understory of shrubs (de Almeida and Riekerk, 1990). Patil (1995) reported

data on water consumption at sites in India, which although high in Eucalyptus, was the

most efficient in terms of water consumed per gram of biomass produced (see also Silva
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et al 2004). In fact, water use efficiency in Eucalyptus actually increases with greater

water availability (Stape et al, 2004a, 2004b). Patil (1995) also noted that there were no

hydrological impacts of Eucalyptus on adjacent crops at these sites. White (1995) stated

that large plantings of Eucalyptus may reduce water yield and lower water tables but this

varies from one situation to another and most importantly can be mitigated through

management practices such as changes in tree stocking regimes. The environmental

considerations of Eucalyptus are the same as those for agricultural crops. Davidson

(1995) noted that drawing water from shallow or deep wells to supply high water

demanding crops such as rice or cotton can have a greater impact on drawing down water

tables than fast growing tree plantations. He also concluded that many potential adverse

effects are reversible, as noted earlier by Poore and Fries (1985).

A review of the environmental issues of Eucalyptus plantations in Brazil was published

by Oak Ridge National Laboratory (Couto and Betters, 1995). This report summarized

that the hydrology of Eucalyptus plantations was comparable to other tree plantations or

natural forest cover and that any effects would largely depend on management practices.

Numerous studies demonstrate that forest cover and any changes in this alter water yield:

reducing forest cover typically increases water yield and vice versa (Bosch and Hewlett,

1982; Sahin and Hall, 1996).

More recent reviews support the points made above. Binkley and Stape (2004) contend

that very large tree plantations must address similar issues of sustainability as seen in

agriculture. They refer to the many hundreds of trials that have been conducted in Brazil,

with particular reference to a very large watershed project conducted in collaboration

with Aracruz Cellulose Company (reported by Almeida et al 2007, described in more

detail below). Binkley and Stape conclude that in semi-arid environments afforestation

with any species of trees may increase water use, lower ground water levels and reduce

streamflow. Given appropriate silvicultural management however, wood production

should face no barriers to sustainability.

Whitehead and Beadle (2004) provided a comprehensive review of the physiological

regulation of water use in Eucalyptus. These species have evolved several mechanisms

to allow them to cope with drought conditions in their native habitats. These include

dynamic changes in leaf area index (LAI), arrangement of leaves, high stomatal

sensitivity to air saturation deficit, osmotic manipulation to maintain turgor in leaves, as

well as an ability to form deep roots. Maximum potential rates of photosynthesis are high

in Eucalyptus compared to other broad-leaved trees, but actual rates are often much less

because of water limitations. Some examples are noted where Eucalyptus plantings have

led to reductions in yields of water catchments. Conversely, the high water usage by

Eucalyptus may be valuable in purposefully lowering water tables to reduced potential

salinity problems. It is therefore important to assess productivity and water use in

relation to climate variables, nutrient supply and options for silvicultural management,

and careful matching of species to sites where available water may be limited. One of the

physiological responses of Eucalyptus to limited water noted by Whitehead and Beadle is

to reduce LAI, thus although Eucalyptus are evergreen species there can be large seasonal

changes in LAI in response to dry seasons. Similar observations were made in reduced

LAI along a gradient of water availability by Ares and Fownes (2000). The root systems
Page 52 of 98

of Eucalypts are dimorphic, with widely spreading lateral root systems below the surface

plus a deep tap root system. In a plantation of 7 year old Eucalyptus trees in Brazil the

tap root extended to a depth of about 2.5m (Almeida and Soares, 2003) consistent with

other observations (see Srivastava et al, 2003). When artificially stressed, by using

plastic sheets on the soil surface to prevent rain entering the soil, young Eucalyptus

developed roots 8 m or greater in depth. Under other conditions water was utilized from

soil below the root zone by upward movement from wetter levels. In considering these

physiological adaptations Whitehead and Beadle conclude that in the case of South

Africa, where planted Eucalyptus replaced native grasslands, the decreased water yields

resulted from increased transpiration in the evergreen and deep rooted Eucalyptus during

the dry season compared to the seasonally dormant grasses. It is well established that

forests have greater evapotranspiration than grasslands (Zhang et al, 1999).

Specific Hydrology Issues for Planted Eucalyptus:

Eucalyptus Afforestation and Hydrology.

In those cases where Eucalyptus has been shown to have negative impacts on hydrology

this has been associated with afforestation, most notably of lands where trees were

previously absent. Typically, these are areas of low rainfall that are normally dominated

by grasses. Under these conditions afforestation with different species of trees, including

Eucalyptus, has lead to changes in the water balance including lowering of water tables

and restricting stream flows. Calder and colleagues have published several reports on

afforestation efforts in India including examples where deep-rooted Eucalyptus were able

to tap into water resources not previously utilized by short-rooted species (Calder et al ,

1997), but also describes cases where water use by Eucalyptus was comparable to

indigenous forests at some sites (Calder, 1994). Similar studies of native grasslands have

documented negative impacts of Eucalyptus on the water balance in South Africa (Lesch

and Scott, 1997; Scott and Lesch, 1997; Scott et al, 1998) and Argentina (Jobbagy and

Jackson, 2004; Engel et al, 2005; Nosetto, 2005). In many of these cases other

introduced trees including pines had similar impacts and particularly in South Africa

impacts on water balance result from a wide variety of introduced species (Le Maitre et

al, 2000; 2002).

These examples contrast with the experience in Brazil where there has been extensive

reforestation with Eucalyptus over many decades. Much of this literature is in

Portuguese but often abstracts are published in English. Lima and colleagues have

published a number of reports that analyzed potential impacts of both Eucalyptus and

pine plantations on the cerrado grasslands in Brazil. Lima et al (1990) showed that in the

region there was adequate rainfall to meet the evapotranspiration demands of Eucalyptus.

A comparison of 6-year old Eucalyptus and pine plantings showed comparable levels of

evapotranspiration during the dry season (May through September) as herbaceous

vegetation (Lima and Freire, 1976). In these trials Eucalyptus actually showed greater

interception than pine (Lima, 1976) and contrasts with references above, but likely

reflected the greater average height of the Eucalyptus at 13.4 m compared to an average

of 6 m for pine.
Page 53 of 98

Similarly, an examination of the water balance of Eucalyptus plantations in China were

not considered to be deleterious for water supplies (Lane et al 2004). While

evapotranspiration exceeded precipitation in the dry season, water storages were

replenished during the wet season.

Comparison of Water Use by Eucalyptus with other Tree Species.

In addition to the reports cited above many authors have concluded that the hydrological

impacts of Eucalyptus are comparable to and should be viewed in the context of other

tree species (see for example Myers et al, 1995; Wullschleger et al, 1998).

One of the largest studies comparing Eucalyptus and native trees conducted to date has

been a catchment area in Brazil of over 280 hectares (owned by Aracruz Cellulose S.A.)

consisting of 190 ha of planted hybrid Eucalyptus and almost 90 ha of native Atlantic

rainforest, that was analyzed over a period of six years. Average precipitation at this site

was 1147 mm, which is similar or less than the sites listed in this permit (range from

~1160 mm in Glades County Florida to almost 1750 mm in Escambia County, Alabama).

Mean high temperature at the Aracruz site was 32.6 C (~91 F) for February (the summer

season in the southern hemisphere), again, comparable to mean high temperatures in the

summer for the sites in this permit. Data from the studies of this catchment area in Brazil

indicated that evapotranspiration was strongly influenced by precipitation (Almeida et al,

2007). In an unusually dry year evapotranspiration was about half that compared to when

water was readily available. In this dry year evapotranspiration exceeded precipitation

but conversely in wetter years evapotranspiration was much less than precipitation. Over

the length of the study evapotranspiration was ~95% of precipitation. This adjustment in

response to varying conditions and water availability was indicative that these hybrid

trees exert strong stomata control and utilize water according to its availability. In a

series of studies in this same area conducted over a period of 8 years Almeida and Soares

(2003, text in Portuguese with abstract and figure legends in English) examined a number

of other hydrological parameters. Stomatal conductance was steady over several months

with adequate water and then dropped significantly as available water dropped and the

predawn leaf water potential (Ψ) increased, again demonstrating strong stomatal control.

Rainfall interception by the Eucalypts averaged ~11% compared to ~24% in the native

forest and water availability (at a depth of 2.5m) is almost identical in the native forests

and the Eucalyptus plantations during the wet summer months but is less in the area with

Eucalyptus during the drier winter. The authors attribute this to the deeper roots systems

(>5m) of the native trees accessing water at deeper levels, while the Eucalyptus (with

roots only to ~2.5m) are limited to the available water in the shallower levels. Finally,

the authors compared the ratio of evapotranspiration and precipitation (ET/P) of the

planted Eucalyptus with the native forest. In years with normal precipitation ET/P was

comparable for both the Eucalyptus and native forest. In years with less than normal

precipitation the native forest had higher ET/P (that is, evapotranspiration was much

greater than precipitation) compared to the Eucalyptus. Based on their data, the authors

suggest that the native forest has a greater consumption of water relative to the

growth/harvest cycle of Eucalyptus, since in the first few years after planting

transpiration in the plantation is much less than the native forest.
Page 54 of 98

Competition for Resources between Eucalyptus and Adjacent Crops

There has been speculation that water use by Eucalyptus could have a negative impact on

water resources available for adjacent vegetation or agricultural crops. Such issues have

been extensively researched in relation to widespread agroforestry systems (reviewed by

Nuberg 1998, and Schroth, 1999). There are important tradeoffs between the positive

effects from windbreaks and shelter belts versus potential competition for light, nutrients

and water resources. Such effects typically occur within 1 to 2 tree-heights (50 to 100

feet for a 50 foot tall tree, Nuberg, 1998) and can be attributed to direct competition by

roots for available soil moisture. Often this can be managed by root pruning to reduce

the area occupied by the tree roots. Impacts attributed to Eucalyptus depend on specific

site conditions, and as with other concerns, there are examples where no negative impact

on adjacent agricultural crops were observed (e.g. Patil, 1995). At some sites this could

be attributed to deeper rooted trees versus shallow rooting crops utilizing water from

different soil profiles. Finally, tree planting has been proposed as a mitigation strategy

where rising water levels increase salinity and reduce crop yields (Hatton and George,

2001).

Summary and Conclusions:

Many studies report that water use in Eucalyptus is comparable to other tree species.

There are some cases where afforestation with Eucalyptus (or other tree species) has lead

to reduced water run-off and supply of streams or changes in water table levels,

especially in regions with limited rainfall. However, in many well documented cases

Eucalyptus plantations do not have any significant negative impacts on hydrology.

Where there have been purported negative impacts, these often reflect more complex

issues such as socioeconomic and land ownership disputes rather than the physiology of

Eucalyptus itself. A key finding of many experiments has been that Eucalyptus is highly

effective in regulating its water consumption relative to available supplies and regulates

its growth accordingly. Based on numerous comparisons that have been made between

the potential hydrological impacts of Eucalyptus and other tree species, we do not expect

that the Eucalyptus trials planted under these permits would be any more impactful on

local hydrology than planting other fast growing trees species.

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APPENDIX III: USDA Forest Service assessment of impacts 

on hydrology

This document was prepared by C. R. Ford and J. M. Vose in response to the document 

titled “Hydrology considerations for planted Eucalyptus” submitted by ArborGen LLC to 

USDA APHIS BRS in support of consideration for approval of permits for field trials of 

Eucalyptus at multiple sites 

Executive Summary

We reviewed the materials provided by ArborGen and synthesized the literature on water 

use by Eucalyptus and other vegetation in the southeastern US. Based on these materials 

and our best professional judgment, we provide the following assessments: 

1. Water use efficiency (WUE) is not a good metric to evaluate impacts on hydrology

From a hydrologic standpoint, total water use (transpiration + interception) is a 

more appropriate metric to assess hydrologic impacts. A species may have high 

WUE (defined by ArborGen as volume of wood produced per amount of water 

required), but still transpire and intercept a significant amount of water. 

2. Annual Et losses by Eucalyptus hybrid plantations planted in the southeast US will 

greatly exceed Et by other native southeastern forest types 

Our review of the literature and estimate of Eucalyptus transpiration suggests that 

water use is at least 2-fold greater than most other native forests in the 

southeastern US. 

3. If Eucalyptus invades native forests, forest water use will increase

Due to a combination of physiological and structural characteristics, Eucalyptus

will use more water than most native species regardless of whether it is planted or 

invades native forests.

4. Afforestation from existing vegetation into Eucalyptus plantations reduces stream 

flow more so than afforestation to pine plantations

Our review of the literature suggests that stream flow will be about 20% lower in 

Eucalyptus plantations vs. pine plantations. 

5. Planting Eucalyptus hybrid plantations will lower the water table, and affect 

groundwater recharge and stream flow dynamics

The combination of shallow and deep roots typical of Eucalyptus species has the 

potential to impact both surface and groundwater hydrology.

6. It is unlikely that lower stocking levels will be an acceptable management practice 

to reduce hydrologic impacts of Eucalyptus plantations.
Page 60 of 98

High biomass production requires fully stocked stands. Reducing stocking to 

minimize hydrologic impacts is likely to counter the benefits of planting fast 

growing Eucalyptus.

Possible impacts of Eucalyptus hybrid plantations on 

southeastern US hydrology

Water use efficiency (WUE) is not a good metric to evaluate impacts on hydrology 

From a physiological standpoint, water use efficiency is defined as the ratio of the moles 

of carbon fixed to the moles of water lost. WUE is a leaf-level metric. The Hydrology 

document prepared by ArborGen provided a ratio of liters of water consumed to grams of 

biomass produced. While these ratios provide good information regarding the 

transpirational cost of biomass production, they do not incorporate information on the 

magnitude of evapotranspirational losses, nor do they integrate stand management effects 

(e.g., planting density, rotation age). Hence, WUE is a poor metric to evaluate the effects 

of Eucalyptus on water resources. For example, Eucalyptus can produce more biomass 

per unit water consumed than native southeastern pines; however, their extremely rapid 

biomass production has proportionally higher transpirational costs and hence greater 

water use. Better metrics of evaluating the impacts of Eucalyptus hybrid plantations on 

hydrology exist. In order of scale, these are evapotranspiration (ET, mm H2O yr-1

), 

transpiration (Et, mm H2O yr-1), and whole-tree water use (Q, kg H2O day-1). 

Evapotranspiration (ET) integrates water loss by Et, interception (Ei), and soil 

evaporation (Es), and is often estimated at the landscape scale using precipitation input 

minus stream flow output on paired-catchments (P-Ro). The net effects of greater

evapotranspiration losses are reduced soil moisture, reduced groundwater depth and 

recharge, and reduced stream flow. These parameters can also be used to evaluate 

impacts on hydrology.

Annual Et losses by Eucalyptus hybrid plantations planted in the southeast US will 

greatly exceed Et by other southeastern forest types 

Previous studies have quantified annual Et from various southeastern US forested and 

crop lands (Table 1). Native pine plantations consume nearly twice the water consumed 

by longleaf pine savannas, but only marginally more than mature upland hardwood 

forests. In contrast, a mature Eucalyptus plantation (age 5, 1111 trees ha-1, LAI of 6 m2

m-2

) growing in southwest GA could potentially transpire 882 mm yr-1, exceeding all 

other forest types on average by a factor of 2.5. The Hydrology document prepared by 

ArborGen states that the proposed sites are on land previously managed for forestry, 

agricultural production or maintained as pasture land. In these cases, we may expect 

Eucalyptus Et to exceed that of previous pine plantations by a factor of 1.6, and previous 

pasture land by a factor of 3.5. The comparison with agricultural crops is more variable; 

Eucalyptus Et may be greater or lesser than crop Et, depending on the crop, the growing 

season, and the management practices.
Page 61 of 98

Table 1

Vegetation type

Mean 

transpiration 

(mm yr-1) Reference

Longleaf pine savanna 244 (Ford et al. 2008)

Old field 250 (Stoy et al. 2006)

Oak-pine-hickory forest 278 (Oren and Pataki 2001)

Upland oak forest 313 (Wullschleger et al. 2001)

Mixed pine hardwood 355 (Phillips and Oren 2001)

Mixed pine hardwood 442 (Stoy et al. 2006)

Planted loblolly pine 490 (Stoy et al. 2006)

Mixed pine hardwood 523 Schafer and others 2002

Slash pine flatwoods 563 (Powell et al. 2005)

Eucalyptus hybrid plantation 882 Estimated for SW GA in average climate 

and rainfall year from model published in 

(Mielke et al. 1999)

Cotton (non-irrigated, annual) 392 (Howell et al. 2004)

Strawberries (irrigated, 7-

month crop, 5-month fallow)

1397 (Clark 1994, Allen et al. 1998)

Watermelon (irrigated, 3-

month crop, 9-month fallow)

237 (Allen et al. 1998, Shukla et al. 2007)

Eucalyptus has much higher stomatal conductance (gs) in humid environments 

compared to native species

The Hydrology document prepared by ArborGen states that Eucalyptus has evolved 

several mechanisms that allows it to cope with drought conditions in their native habitats, 

including high stomatal sensitivity to air vapor pressure deficit (VPD). Across many taxa 

of plants, two main gs responses to VPD exist (Figure 1). Both strategies regulate gs (and

thus transpiration) according to allowable variation in leaf water potential. The benefit of 

having a high δ, is having a high conductance in humid environments (gsref). The 

southeastern US is a relatively humid environment, with average daily VPD values 

around 1.5 kPa (Ford et al. 2004). In this humid environment, we can expect that 

Eucalyptus hybrid plantations will have stomatal conductance rates that are roughly 

double the conductance rates of native southeastern pine species. This is one mechanism 

that confers a greater transpiration rate of the former compared to the latter. 
Page 62 of 98

Figure 1: Stomatal 

conductance response to 

VPD for pine and Eucalyptus

(Oren et al. 1999)

If Eucalyptus invades native forests, forest water use will increase

In general, species that evolved in arid climates have a more sensitive gs response to VPD 

than those in more mesic environments (McDowell et al. 2008). When comparing water 

use of native species to plant species that have invaded a system (invasives), a recent 

meta-analysis across all biomes shows that stomatal conductance and photosynthesis are 

significantly greater in the invasive species compared to any of the native species in the 

system (Cavaleri and Sack 2008). Specifically, for systems that had been invaded by tree 

life-forms, stand level transpiration was significantly greater compared to un-invaded 

systems (Cavaleri and Sack 2008).

Afforestation from existing vegetation into Eucalyptus plantations reduces stream

flow more so than afforestation to pine plantations

The Hydrology document prepared by ArborGen states that afforestation with any tree 

species may reduce stream flow; and that while Eucalyptus plantations might reduce 

stream flow, the reduction would be comparable to the reduction by pine plantations. 

Recent research suggests that Eucalyptus plantations would reduce stream flow more 

than pine plantations, and more importantly, Eucalyptus plantations could eliminate low 

flows. In a review of more than 20 catchment conversion studies, Farley and others 

(2005b) showed that converting existing vegetation to Eucalyptus plantations reduced 

stream flow by 20% more than converting it to a pine plantation. This review also 

VPD (kPa)

0 1 2 3 4 5 6

gs (mmol m-2 s-1)

0

100

200

300

400

500

600

700

800

Eucalyptus

δ = -194

gs

ref = 328

Pine

δ = -81

gs

ref = 144
Page 63 of 98

showed that the loss of low flows were more complete for Eucalyptus plantations 

compared to pine plantations (100% vs. ~80% reduction of low flows).

In perennial streams throughout the southeast which have base flows sustained by 

subsurface flow from the water table (or unconfined aquifers), elimination of low flows 

may have important ramifications for threatened and endangered aquatic species, such as 

the gulf strain striped bass, and thee species of endemic freshwater mussels (Golladay et 

al. 2004b, Couch and McDowell 2006a).

Planting Eucalyptus hybrid plantations will lower the water table, and affect 

groundwater recharge and stream flow dynamics

The Hydrology document prepared by ArborGen states that afforestation with any tree 

species may lower ground water levels. This is highly dependent on subsurface flow 

patterns, local hillslope hydrology, and species-specific rooting patterns. For example, in 

sites where the water table can be recharged laterally, if roots extend to the water table, 

then stomatal conductance and transpiration can be maintained even when water in the 

upper soil layers is insufficient to maintain transpiration. If Eucalyptus hybrid plantations 

mine water from the saturated zone (i.e., water table), groundwater recharge could be 

reduced. The southeastern Coastal Plain is characterized in many places by karst geology 

in which groundwater from the semi-confined Upper Floridan Aquifer (UFA) is 

hydraulically connected to the water table (surface water) (Opsahl et al. 2007). Mean 

water table depths typically range 3–8 m (Ford et al. 2008). The mean maximum rooting 

depth for Eucalyptus is 15 m, characteristic of its dimorphic rooting pattern; in contrast, 

mean maximum rooting depths of pine plantation (P. taeda and P. elliottii) and grass 

species are 3 m and 2.6 m, respectively (Canadell et al. 1996b). The average age of 

groundwater in the UFA is ~20 years (Happella et al. 2006) and groundwater is regularly 

recharged by surface water in this region (Opsahl et al. 2007). Conversion to Eucalyptus

on sites with water tables <10 m will likely lower down-slope water tables via direct 

means (i.e., direct use of ground water by deep roots), affect groundwater-aquifer 

dynamics, and result in ET rates that exceed precipitation input, as have been reported for 

this species in other locations (Calder et al. 1997b).

The Hydrology document prepared by ArborGen states that the high water usage 

by Eucalyptus may be valuable in purposefully lowering water tables to reduced potential 

salinity problems. Eucalyptus has been used to afforest areas and lower the saline 

groundwater in highly weathered landscapes (e.g., AUS). This application is not relevant 

to the southeastern US, as soils are not saline. Furthermore, receding groundwater levels 

in the UFA are being replaced in costal areas by saltwater (i.e., saltwater intrusion) 

(Andersen et al. 2006). Thus, lowering the water table, and the groundwater levels in the 

UFA would not reduce salinity problems (as stated in the Hydrology document), and may 

actually exacerbate them. 

Key Point: The significance of the impact on groundwater and stream flow will depend 

greatly on the area extent, size, and spatial distribution of the plantations. For 

example, a few small (i.e., < 10 ha) and well dispersed plantations may only have very 

localized impacts and negligible impacts at the watershed scale.
Page 64 of 98

Management of Eucalyptus as coppice stands will affect water use of future rotations

Management practices may create a perennial root stock in Eucalyptus plantations. If 

Eucalyptus plantations are managed as coppice stands, the remaining mature, deeplypenetrating root stock may be able to supply the second rotation stems with more water 

resources for use than similar sized stems in their first rotation (Swift and Swank 1981). 

It is unlikely that lower stocking levels will be an acceptable management practice to 

reduce hydrologic impacts of Eucalyptus plantations

Some of the reports cited in the ArborGen document discuss the potential for altering 

management practices to minimize the impacts of intensively managed Eucalyptus

plantations on hydrology. The most viable option for reducing hydrologic impacts is to 

manage stocking (“stocking” is a term to describe the how much of the site is occupied 

by the species of interest; stand basal area expressed in m2 stem area hectare-1 is often 

used as a measure of stocking). Water use is highly regulated by stand leaf area and 

reducing basal area will result in lower stand leaf area. Empirical research (Douglass and 

Swank 1972) at Coweeta has shown that stand basal area needs to be reduced by at least 

15 % before any impact on stream flow is obtained. Large and sustained increases in 

stream flow typically require significant reductions in stocking because trees growing in 

more open conditions will increase transpiration rates in response to changes in 

micrometeorological conditions in the tree crown. 

From a practical standpoint, it is unlikely that lower stocking levels will be an 

acceptable management practice for Eucalyptus plantations because one of the primary 

objectives of growing Eucalyptus is to maximize biomass production -- this requires fully 

stocked stands. 

Monitoring Impacts on Hydrology

Several options are available to monitor the impacts of Eucalyptus plantations on 

hydrology. Options include direct or indirect measurements of impacts, and vary in 

complexity and cost. The ArborGen document provided basic information on the 

hydrologic cycle and its components so these will not be repeated here. 

Indirect Measurements:

Transpiration & Interception

Instrumentation required = rain gauges, throughfall collectors, sap flow sensors

PROS: direct measure of change in water use component on hydrologic cycle

CONS: does not directly measure impacts on stream flow or groundwater; 

expensive, high maintenance

Soil Moisture

Instrumentation = TDR probes and data loggers (automated); soil probe for 

gravimetric (manual)

PROS: easy to implement, relatively inexpensive

CONS: does not directly measure impacts on stream flow or groundwater
Page 65 of 98

Direct Measurements:

Groundwater Depth

Instrumentation = access wells & pressure transducers (automated); access wells 

(manual)

PROS: direct measure of impacts; reliable

CONS: expensive 

Stream flow

Instrumentation = flumes & data loggers (automated); pressure transducer, rating 

curve, stream survey

PROS: direct measure of impacts; reliable

CONS: expensive, requires stream, expertise

Regardless of the monitoring approach chosen, the monitoring design will require 

a suitable control for comparison. Ideally, the plantation site(s) and the control site(s) 

would be measured for at least 1 to 2 years prior to be being planted, and then both sites 

would be measured for the duration of the monitoring period. We recommend that the 

monitoring period begin at plantation establishment and continue through canopy closure 

(approximately 5 years). 

Additional analyses on groundwater dynamics and linkages with aquatic ecosystems 

are required

We recommend that APHIS solicit input from experts on groundwater hydrology (e.g., 

from USGS) to assess the potential impacts on groundwater recharge and associated 

dynamics. In addition, our analysis suggests that stream flow will be reduced by at least 

20% relative to pine plantations and perhaps even greater reductions will be observed 

relative to native ecosystems. We recommend that APHIS solicit input from aquatic 

ecologists to assess the potential impacts on aquatic ecosystems and associated species. 
Page 66 of 98

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computing crop water requirements. FAO - Food and Agriculture Organization of

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greater than rainfall input-a possible explanation from southern India. Hydrology

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mussel assemblages (Bivalvia:Unionidae) to a record drought in the Gulf Coastal

Plain of southwestern Georgia. Journal of the North American Benthological

Society 23: 494-506.

Happella JD, Opsahl S, Top Z, Chanton JP. 2006. Apparent CFC and 3H/3He age

differences in water from Floridan Aquifer springs. J. Hydrol. 319: 410–426.

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and dryland cotton on the northern Texas high plains. Journal of

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9437(2004)130:4(277)

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Williams DG, Yepez EA. 2008. Tansley review: Mechanisms of plant
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succumb to drought? New Phytol. 178: 719–739.

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plantation. Trees- Structure and Function 13: 152-160.

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surface-water exchanges using streamflow difference analyses. Journal of the

American Water Resources Association 43: 1132–1141.

Oren R, Pataki DE. 2001. Transpiration in response to variation in microclimate and soil

moisture in southeastern deciduous forests. Oecologia 127: 549-559.

Oren R, Sperry JS, Katul GG, Pataki DE, Ewers BE, Phillips N, Schafer KVR. 1999.

Survey and synthesis of intra- and interspecific variation in stomatal sensitivity to

Vapour pressure deficit. Plant Cell Environ. 22: 1515-1526.

Phillips N, Oren R. 2001. Intra- and inter-annual variation in transpiration of a pine

forest. Ecol. Appl. 11: 385–396.

Powell TL, Starr G, Clark KL, Martin TA, Gholz HL. 2005. Ecosystem and understory

water and energy exchange for a mature, naturally regenerated pine flatwoods

forest in north Florida. Can. J. For. Res. 35: 1568-1580.

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watermelon in southwest Florida. Institute of Food and Agricultural Sciences

(IFAS), University of Florida. Report No. WRP-LY-0009 submitted to Southwest

Florida Water Management District

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Kim H, Oren R. 2006. Separating the effects of climate and vegetation on

evapotranspiration along a successional chronosequence in the southeastern US.

Global Change Biology 12: 2115–2135.

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and regrowth. Hydrological Sciences Bulletin 26: 245–256.

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deciduous forest as estimated by xylem sap flow techniques. For. Ecol. Manage.

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APPENDIX IV: Threatened and Endangered Species Analysis

The threatened and endangered species analysis was accomplished by considering the possibility of 

effects on all listed species including species proposed for listing and designated critical habitat 

including habitat proposed for designation within the county where the test site is located. 

Normally consideration would be limited to only those species and critical habitat identified by the 

United States Fish and Wildlife Service and/or National Marine Fisheries Service to be likely found 

in the area of the field tests. However, because the locations are declared by the applicant to be 

confidential business information, APHIS instead obtained species lists and critical habitat 

information for the entire county where the field tests are to occur. Although it is very clear that 

there could be no effect on some species (e. g. humpback whale) all species on the list and all 

critical habitat in each county are included in the discussion below. 

The analysis was originally completed in early 2008, and updated in February 2009 to reflect 

changes to species’ regulatory status. After receiving comments in response to the draft EA, it was 

decided to review the entire analysis to ensure that it was up to date and satisfies the requirements 

of Section 7 (a) (2) of the Endangered Species Act. This was completed in April 2010. Species 

lists and critical habitat for each county were verified with USFWS field offices to ensure accuracy. 

The review resulted in the addition of some new species to the county lists, and the removal of 

others from consideration. The Glades County, Florida site was not included in the revised analysis 

because the applicant removed this location from the permit applications. There are no significant 

changes to the potential for species to be affected by the action, or the likelihood of any species to 

be found in the test sites. 

The following resources were used in the analysis:

US Fish and Wildlife Service Endangered Species Homepage: http://www.fws.gov/endangered/

US Fish and Wildlife Service ECOS system: http://ecos.fws.gov/ecos_public/index.do

NatureServe Explorer: http://www.natureserve.org/explorer/index.htm 

Google Earth with critical habitat metadata supplied by US Fish and Wildlife Service 

County species lists from FWS field offices in Alabama, Georgia, Louisiana, Mississippi, South 

Carolina, Texas and Florida.

South Carolina Heritage Trust Database

Telephone contacts with USFWS Field Office personnel

Discussions with property owners of field test sites 

Baldwin County, AL (AL-BAL-01)

This location has been an agricultural research station for more than 20 years. The location has 

been used for managed production of annual agricultural crops and forest trees. Approximately 6.2 

acres of field trials of genetically modified Eucalyptus Hybrid (EH1) trees of some of the lines in 

the permit request are being grown under issued permit # 06-325-111r under which these trees are 

allowed to flower. The oldest of these trees at this site are now entering their fifth growing season. 

An additional 0.75 acres of tests with some of these lines are being grown under permit 08-039-

102rm. Site preparation involved herbicide application, subsoiling, and planting of trees in flat 
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beds. The surrounding areas of the test site consist of field plantings of agricultural crops, 

experimental forest trees and an abandoned pecan orchard.

Fourteen TES animals and one TES plant are listed in Baldwin County. The TES Animals include 

loggerhead sea turtle (Caretta caretta), green sea turtle, (Chelonia mydas), Alabama red-belly turtle 

(Pseudemys alabamensis), Alabama beach deermouse (Peromyscus polionotus ammobates), 

Perdido Key beach deermouse (Peromyscus polionotus trissyllepsis), piping plover (Charadrius 

melodus), least tern (Sternula antillarum), West Indian manatee (Trichechus manatus), wood stork 

(Mycteria americana), Kemp's ridley sea turtle (Lepidochelys kempii), gulf sturgeon (Acipenser 

oxyrinchus desotoi), Alabama sturgeon (Scaphirhynchus suttkusi), heavy pigtoe mussel 

(Pleurobema taitianum), and inflated heelsplitter mussel (Potamilus inflatus). The listed plant is 

American chaffseed (Schwalbea americana). 

The American chaffseed occurs on sandy peat, sandy loam, acidic, seasonally moist to dry soils. It 

is generally found in habitats described as open, moist pine flatwoods, fire-maintained savannas, 

ecotonal areas between peaty wetlands and xeric sandy soils, and other open grass-sedge systems. 

According to Jody Smithen (contacted February 20, 2008) of the Daphne Field Office USFWS, the 

only location this plant is known to be in the county is in the northeast corner, far from the release 

site. The plant has no designated critical habitat. 

The four turtle species (loggerhead sea turtle, green sea turtle, Alabama redbelly turtle and Kemp’s 

ridley sea turtle), the gulf and Alabama sturgeons, and the West Indian manatee occur in aquatic 

habitats, and their habitat systems (bays, lagoons, salt marshes, creeks, ship channels, and other 

saltwater and freshwater environments) do not overlap with the trial site. The wood stork primarily 

inhabits wetland systems notably cypress or mangrove swamps and would not use the field test site. 

The two mouse species listed above (Alabama beach mouse and Perdido Key Beach deermouse) 

are found only in coastal dune areas and Perdido Key Beach, respectively, where they feed on sea 

oats, bluestems, and a variety of insects. Both habitats are located over 60+ miles from the 

proposed field trial. The piping plover uses sparsely vegetated dunes and coastal beaches in 

southern Baldwin County, also far away from the field site (about 60 miles). The Least Tern 

breeds on seacoasts, beaches, bays, estuaries, lagoons, lakes, and rivers and rests on sandy beaches, 

mudflats, and salt-pond dikes which are far away from the field test site. 

Three species have been removed from the county list since the original TES effects analysis was 

completed in 2008. The FWS has determined that the species are so unlikely to be found in the 

county that it would be unreasonable for federal agencies to include them in TES analysis for 

projects within the county. These are the red-cockaded woodpecker (Picoides borealis), the

reticulated flatwoods salamander (Ambystoma bishopi), and the Eastern indigo snake (Drymarchon 

corais couperi). However, we are retaining the information and findings from the earlier analysis 

as follows: The red-cockaded woodpecker could potentially visit the field tests but this species 

prefers mature pine stands as habitat for nesting. The reticulated flatwoods salamander uses the 

wet pine flat-woods associated with ephemeral wetlands. They typically breed in the low wetlands 

where they lay their eggs. After hatching, they spend 11 to 18 weeks as larvae before 

metamorphosing into adults and leaving the wetland for higher ground to burrow into the soil. It is 

important that the area through which the species moves is vegetated with grasses. Reticulated 

flatwood salamanders may move as far as 450 meters from their breeding sites. The area 
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surrounding the release site out to beyond this distance has been intensively managed for over 20 

years as an agricultural research station, making it unlikely that the species would find this habitat 

useable. The Eastern indigo snake is known to inhabit a wide range of habitats (agriculture fields, 

pine flat-woods, wet depressions, stream bottom thickets and margins of swamps). It appears to be 

very rare in Baldwin County where a case has been reported in an unknown location (US Forest 

Service). According to Jody Smithen (contacted February 20, 2008) of the Daphne Field Office 

USFWS, the species has not been documented in the county for many years, but there are 

occasionally unsubstantiated reports. They do not feel this field trial poses concern for this species. 

Although it is highly unlikely that the species would be found at the site, the applicant will provide 

all workers with identifying characteristics of the snake and instructions on what to do if the species 

is encountered. These measures are a variation of standard protective measures the USFWS uses 

when they have reached a “may affect” determination for construction sites.

Critical Habitat: Most of the TES animals within the county area use inshore or wetland systems 

most of which are concentrated essentially in the southern and southeastern coastal beaches of 

Baldwin County. There is critical habitat listed for the Perdido Key beach deermouse and the 

piping plover. Notice of designation of critical habitat for the reticulated flatwood salamander 

(final rule) was published in the Federal Register on February 10, 2009 

(http://edocket.access.gpo.gov/2009/pdf/E9-2403.pdf). There is no designated critical habitat for 

this species in Alabama. The closest critical habitat (for the Perdido Key Beach deer mouse and the 

piping plover) is about 65 miles away. The gulf sturgeon has critical habitat in Alabama but this 

does not occur in Baldwin County.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

 

Escambia County, AL (AL-ESC-01)

This location had previously been used as an intensely managed pasture for more than 5 years and 

was planted with grasses suitable for cattle grazing. Site preparation involved herbicide application 

to remove existing grasses, subsoiling, preparation for possible irrigation, and planting of the test 

trees in flat beds. Field tests with some of the lines in these permit requests were established in 

2007 (0.3 acres) and 2008 (0.2 acres) and are currently covered under permit 08-039-102rm. The 

surrounding areas of the test site consist of experimental forest trees, ~ 30 year-old slash pine and a 

re-forested area with less than 7 year-old mixed stands of pine and hardwood species.

Three TES animals are listed for Escambia County. The animals are gulf sturgeon, (Acipenser 

Oxyrinchus desotoi), red-cockaded woodpecker (Picoides borealis) and wood stork (Mycteria 

americana). The gulf sturgeon occurs in the Gulf of Mexico and spawns in freshwater rivers. It 

will not be affected by the field test since the closest river is over 3.5 miles away. The red 

cockaded woodpecker inhabits old growth forests, primarily longleaf pine. It could visit the field 
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test site but would not nest there. The wood stork primarily inhabits wetland systems notably 

cypress or mangrove swamps and would not find the field test site hospitable.

Critical Habitat: The designated critical habitat for the gulf sturgeon includes the Escambia River 

System in Santa Rosa and Escambia counties, Florida and Escambia, Conecuh, and Covington 

counties, Alabama. The establishment of the field test site would not impact this habitat. It is 

about 3.5 miles away from the Conecuh River.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Evans County, GA (GA-EVA-01)

This location has been a commercial nursery for forest seedling production for over 30 years. Site 

preparation involved herbicide application, subsoiling and planting of trees in flat beds. A field test 

of 0.2 acres of some of the lines included in these permit requests were established under permit 

08-039-102rm in 2008. The areas surrounding the test site consist of nursery beds of forest tree 

seedlings, experimental forest trees, agricultural crops and mixed stands of hardwood and pine.

There are four TES animals listed for Evans County. There are no listed plants. The animals are 

the Eastern indigo snake (Drymarchon corais couperi), wood stork (Mycteria americana), redcockaded woodpecker (Picoides borealis), and frosted flatwoods salamander (Ambystoma 

cingulatum). 

The listed birds (red-cockaded woodpecker and wood stork) could potentially visit the field test site 

but would not nest there. The red-cockaded woodpecker nests in old growth longleaf pine. The 

wood stork nests in marshes, floodplain lakes, and swamps. The flatwoods salamander (Ambystoma 

cingulatum) was a species that required further analysis because it typically inhabits longleaf or 

slash pine forests lying between drier land upslope and wetlands and seasonally inhabits wet pine 

flat-woods with vernal pools. Originally it was associated with a unique community of longleaf 

pine/wire grass, but much of this habitat is now replaced by slash pine plantations. According to 

Kathy Chapman (contacted February 22 and 26, 2008) of the Coastal Georgia Field Office, there 

should not be an effect on the species because there would be no change to the habitat. However, 

she suggested verifying that there is no suitable breeding habitat near the release site. She provided 

information on life history and suitable habitat for the species. Viewing the site using Google Earth 

does not readily identify any suitable breeding habitat. To ensure that there is no suitable habitat in 

the area, BRS provided the species information to the applicant and in early March 2008, the 

applicant conducted a breeding habitat survey of the area within 450 meters (1476 feet) of the 

release site. No suitable habitat was found. For the eastern indigo snake (Drymarchon couperi) 

according to Kathy Chapman (contacted February 22 and 26, 2008) of the Coastal Georgia Field

Office, the species is associated with the gopher tortoise in sandhill areas with wetlands. However, 
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land use is a determining factor on the likelihood of either species being present. The site has been 

used as a commercial nursery for forest seedling production for over 30 years. She indicated that 

she was not concerned with the site because there would be no change to the habitat. Although it is 

highly unlikely that the species would be found at the site, the applicant will provide all workers 

with identifying characteristics of the snake and instructions on what to do if the species is 

encountered. These measures are a variation of standard protective measures the USFWS uses 

when they have reached a “may affect” determination for construction sites.

Critical Habitat: There is no designated critical habitat or habitat proposed for designation in the

county.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

St. Landry’s Parish, LA (LA-SLP-01)

This location has been an experimental agricultural farm for more than 25 years, used for 

conducting research experiments with soybean, cotton and wheat. Site preparation will involved 

herbicide application, subsoiling and planting of trees in flat beds. Field tests of some of the lines 

included in these permit requests were established during 2006, 2007 and 2008 on 1.9 acres total, 

now covered under permit 08-039-102rm. Approximately 1.7 acres were subsequently terminated. 

The areas surrounding the test site consist of agricultural fields of rice, sugarcane and millet.

There are two TES animals listed for St. Landry’s Parish; the pallid sturgeon (Scaphirhynchus

albus) and the Louisiana black bear (Urus americanus luteolus). There are no listed plants.

The pallid sturgeon occurs in larger channels of the Mississippi-Missouri river system. The field 

test site is approximately 24 miles away from the Mississippi river, so would not be impacted by 

the field test. The Louisiana black bear (Ursus a. luteolus) prefers bottomland forests with diverse 

food resources, including a variety of hard-mast-producing species. Its habitat includes remote 

areas with little or no human activity so it would not likely be found at the site.

Critical Habitat: There is no designated critical habitat or habitat proposed for designation in the

parish. The Louisiana black bear does have critical habitat in adjoining Pointe Coupee parish, but 

this is over 10 miles from the release site. 

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 
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designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Marshall County, MS (MS-MAR-01)

This location has been an agricultural research station for more than 50 years, used for conducting 

research experiments with agricultural crops and grasses. The test site was previously used for 

experimental planting of grasses. Site preparation involved herbicide application, subsoiling and 

planting of trees in flat beds. A field test of 0.5 acres of some of the lines included in these permit 

requests were established in 2007, now under permit 08-039-102rm. This test was subsequently 

terminated. The areas surrounding the test site consist of experimental forest trees, agricultural 

fields, and less than 5 year-old pine plantations.

There are no listings for TES animals or plants in the county. 

Critical Habitat: There is no designated critical habitat or habitat proposed for designation in the

county.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. Field activities will result in no changes to the habitat used 

by any listed species or species proposed for listing. The site is not within or near designated 

critical habitat or habitat proposed for designation. Therefore, the action will have no effect on 

listed species or species proposed for listing and would not affect designated critical habitat or 

habitat proposed for designation.

Pearl River County, MS (MS-PRC-01)

This location has been used as an agricultural research station for more than 5 years for conducting 

research experiments with agricultural crops and grasses. The test site was used for experimental 

planting of grasses. Site preparation involved herbicide application to remove existing grasses, 

subsoiling, preparation for irrigation installation, and planting of trees in flat beds. Field tests of 

some of the lines included in these permit requests were established in 2007 (2.0 acres) and 2008 

(0.98 acres), now covered under permit 08-039-102rm. The areas surrounding the test site consist 

of a grape research farm, mixed stands of hardwoods and pine, and a residential area.

There are five TES animals listed in this county and one TES plant. The animals are the ringed 

map turtle (Graptemys oculifera), gopher tortoise (Gopherus polyphemus), Louisiana black bear 

(Ursus a. luteolus), gulf sturgeon (Acipenser oxyrhynchus desotoi) and inflated heelsplitter 

(Potamilus inflatus). The listed plant is Louisiana quillwort (Isoetes louisianensis). 

The ringed map turtle (Graptemys oculifera) inhabits wide rivers with strong currents, adjacent 

white sand beaches, and an abundance of basking sites in the form of brush, logs, and debris. The 

field test will not impact this aquatic species which occurs in the Pearl River system –

approximately 11 miles from the field test site. The gopher tortoise (Gopherus polyphemus) 

inhabits dry sand ridges dominated by pine and areas maintained by fire. It is common in longleaf 

pine forests, but its numbers have decreased with the replacement of longleaf pine forests with 
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loblolly pine forests. This field test is located in an agricultural research station that would be an 

inhospitable environment for the gopher tortoise. According to James Harris (contacted February 6 

and 7, 2008), Supervisory Wildlife Biologist with the USFWS in Lacombe, LA, the species is 

found in a large geographic area that includes the release site. However, the species is not found 

everywhere within this geographic area. The species is not likely to be on the site because of its 

location on a facility used for many years as an agricultural research station. The research facility 

was contacted to determine if gopher tortoises have been observed at the facility. The farm 

manager has not observed the species at the facility, and no sightings have been reported to him 

during his nine years as manager. He is familiar with the species and has seen them at another 

location about 7-8 miles from the release site. Another employee contacted has worked at the site 

for over 35 years and has never seen a gopher tortoise at the facility but did observe one 

approximately ten years ago about 1⁄4 mile from the facility. The applicant surveyed the site for the 

presence of gopher tortoise burrows on January 29, 2008 and none were found. Considering the 

use of the facility, testimony of the facility employees, and the negative result of the survey, it can 

be concluded that the species is not present now and would be unlikely to use the site while it 

operates as an agricultural research station. The Louisiana black bear (Ursus a. luteolus) prefers 

bottomland forests with diverse food resources, including a variety of hard-mast-producing species. 

Its habitat includes remote areas with little or no human activity so it would not likely be found at 

the site. The gulf sturgeon, the inflated hellsplitter mussel, and the Louisiana quillwort occur in 

aquatic environments so would not be affected by the field test. 

Critical habitat: The gulf Sturgeon has designated critical habitat in this county in the Pearl River 

system. The Pearl River is about 11 miles from the field test site. Tributaries of the Pearl River 

system are approximately 3.5 and 7.7 miles from the test site. 

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Bamberg County, SC (SC-BAM-01)

This location has been a managed forest plantation for more than 14 years. The location has been 

specifically used for short-rotation planting of hardwoods and softwood trees for forestry research. 

Standard silvicultural practices for site preparation, irrigation, fertilization, planting and harvesting 

have been used at this location. Similar practices will be used for the additional field tests to be 

established at this site. Several field tests of some of the lines included in these permit requests (3.5 

acres total) were established from 2006 to 2008. These tests were covered under permit 08-039-

102rm, and all but 0.2 acres were subsequently terminated. The areas surrounding the test site 

consist of experimental forest trees, young pine plantations, mixed stands of hardwoods and pine, 

and agricultural fields.

Three TES are listed in Bamberg County. The TES species listed are a plant, Canby’s dropwort 
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(Oxypolis canbyi), and two animals, the wood stork (Mycteria americana) and the red cockaded

woodpecker (Picoides borealis). 

Canby’s dropwort is an herbaceous perennial whose existing populations are maintained mainly 

through asexual reproduction. This species is strongly clonal, reproducing vegetatively by means 

of stoloniferous rhizomes. It has been found in a variety of habitats, including cypress ponds, 

grass-sedge dominated Carolina bays, wet pine savannahs, shallow pineland ponds and cypresspine swamps or sloughs. The largest and most vigorous populations reported occur in open bays or 

ponds which are flooded throughout most of the year and which have little or no canopy cover. It 

grows in soils with a medium to high organic content, high water table, that are deep, poorly 

drained, and acidic. The Lisa Matthews Memorial Bay is a 52 acre site in Bamberg County which 

was given to the South Carolina Native Plants Society by the Nature Conservancy for the purpose 

of preserving Canby's dropwort (Oxypolis canbyi). This depression wetland is apparently a 

remnant Carolina Bay which is being restored and expanded to protect this endangered species. 

The location of the field test under this permit is a managed forest research area and does not 

provide the proper habitat for the species. 

The wood stork inhabits riparian areas with lagoons and shallow water. It lives in freshwater 

situations: marshes, swamps, lagoons, ponds, flooded fields and brackish wetlands. It nests mostly 

in the upper parts of cypress trees, mangroves, or dead hardwoods over water or on islands along 

streams or adjacent to shallow lakes and feeds in freshwater marshes, swamps, lagoons, ponds, 

flooded pastures and flooded ditches, and in depressions in marshes. The field trial location is 

unsuitable habitat for the wood stork so it will be very unlikely to occur in this location. The red 

cockaded woodpecker inhabits old growth forests, primarily longleaf pine and might visit the field 

test site but would not nest there. 

Critical Habitat: There is no designated critical habitat or habitat proposed for designation in the 

county.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Berkeley County, SC (SC-BER-01 and SC-BER-02)

There are two release site locations in Berkeley County. One site is an extension of a greenhouse 

facility that has been used for acclimatization of transgenic and non-transgenic plants for more than 

7 years. The release site is located adjacent to greenhouse facilities and is surrounded by 

hardwoods and pine plantations. The trees are held here on a temporary basis until field planting. 

The other location has been a managed forest plantation for more than 7 years. The location has 

been specifically used for short rotation planting of cottonwood and Eucalyptus for forestry 

research. Site preparation involved herbicide application, subsoiling, drip irrigation installation, 
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and planting of trees in flat beds. The test site is located adjacent to greenhouse facilities and is 

surrounded by pine plantations. Field tests of some of the lines included in these permit requests 

were established in 2006 (0.3 acres) and 2007 (0.2 acres) that were covered under permit 08-039-

102rm but subsequently terminated.

Five threatened or endangered species (TES) animals and three plants are listed in Berkeley 

County. The TES animal species are shortnose sturgeon (Acipenser brevirostrum), frosted 

flatwoods salamander (Ambystoma cingulatum), red-cockaded woodpecker (Picoides borealis), 

West Indian manatee (Trichechus manatus), and wood stork (Mycteria americana). The TES 

plants are pondberry (Lindera melissifolia), Canby's dropwort (Oxypolis canbyi) and American 

chaffseed (Schwalbea americana). 

The shortnose sturgeon (Acipenser brevirostrum) occurs in rivers and estuaries. The West Indian 

manatee (Trichechus manatus) occurs in shallow coastal waters, rivers, bays and lakes; none of 

which are close to any of these release locations. The red cockaded woodpecker (Picoides 

borealis) inhabits old growth forests, primarily longleaf pine and might visit the field test site but 

would not nest there. The wood stork (Mycteria americana) primarily inhabits wetland systems 

notably cypress or mangrove swamps and would not find the field test site hospitable. The frosted 

flatwoods salamander (Ambystoma cingulatum) inhabits longleaf or slash pine forests lying 

between drier land upslope and wetlands and seasonally inhabits wet pine flat-woods with vernal 

pools. Discussions with Laura Zimmerman (contacted February 14 and 20, 2008) of the Charleston 

Field Office of the USFWS indicate that the species is not known to be in the area of the release. 

Known populations in the county are in the Francis Marion National Forest, far from the release 

site. A check of the SC Heritage Trust Database did not identify any occurrences in the area of the 

release. One site in Berkeley County is a fenced research plot/holding area so it is highly unlikely 

that these animal species would occur at this location. The other site is a managed forest plantation 

for short rotation planting of cottonwood for forestry research. The bird species might visit the 

location but would not nest there. 

Canby’s dropwort (Oxypolis canbyi) is an herbaceous perennial whose existing populations are 

maintained mainly through asexual reproduction. This species is strongly clonal, reproducing 

vegetatively by means of stoloniferous rhizomes. It has been found in a variety of habitats, 

including cypress ponds, grass-sedge dominated Carolina bays, wet pine savannahs, shallow 

pineland ponds and cypress-pine swamps or sloughs. The largest and most vigorous populations 

reported occur in open bays or ponds which are flooded throughout most of the year and which 

have little or no canopy cover. It grows in soils with a medium to high organic content, high water 

table, that are deep, poorly drained, and acidic. The pondberry (Lindera melissifolia) occurs in 

similar locations, in wetland habitats such as bottomland and hardwoods in the interior areas, and 

the margins of sinks, ponds and other depressions in the more coastal sites. The plants generally 

grow in shaded areas but may also be found in full sun. The chaffseed (Schwalbea americana)

occurs on sandy peat, sandy loam, acidic, seasonally moist to dry soils. It is generally found in 

habitats described as open, moist pine flatwoods, fire-maintained savannas, ecotonal areas between 

peaty wetlands and xeric sandy soils, and other open grass-sedge systems. The site in Berkeley 

County is a fenced research plot/holding area which would be very inhospitable to these species. 

Laura Zimmerman (contacted February 14 and 20, 2008) of the Charleston Field Office of the 

USFWS states that she does not believe the species would be likely to be in the area. According to 
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the species’ recovery plan, most known occurrences are on US Forest Service land and the only 

two occurrences on private land are not near the release site. A check of the SC Heritage Trust 

Database did not identify any occurrences in the area of the release. 

Critical Habitat: Notice of designation of critical habitat for the frosted flatwood salamander (final 

rule) was published in the Federal Register on February 10, 2009 

(http://edocket.access.gpo.gov/2009/pdf/E9-2403.pdf). Unit FFS-6 is within Berkeley County. 

This is located in the Francis Marion National Forest which is about 19-20 miles away from the 

release location. 

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at these release sites. If they were to enter the sites, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The sites are not within or near designated critical habitat or habitat proposed 

for designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Charleston County, SC (SC-CHA-01)

This location has been a managed forest plantation for more than 10 years. The location has been 

specifically used for short-term planting of hardwoods and softwood trees for forestry research. 

Standard silvicultural practices for site preparation, irrigation, fertilization, planting and harvesting 

were used at this location. Similar practices were used for the additional field tests established at 

this site. Field tests of 3.1 acres of some of the lines included in the permit request were 

established during 2006 and 2007. These tests were covered under permit 08-039-102rm, and were 

all subsequently terminated. The test plots adjacent to the field test site include young mixed 

stands of hardwoods and pines. No additional trials are currently planned for this site.

Thirteen TES animals and 4 TES plants are listed for Charleston County. The animal species are: 

shortnose sturgeon (Acipenser brevirostrum), West Indian manatee (Trichechus manatus), Kemp's 

ridley sea turtle (Lepidochelys kempii), leatherback sea turtle (Dermochelys coriacea) and green sea 

turtle, (Chelonia mydas), loggerhead sea turtle (Caretta caretta), wood stork (Mycteria americana), 

red-cockaded woodpecker (Picoides borealis), piping plover (Charadrius melodus), frosted 

flatwoods salamander (Ambystoma cingulatum), finback whale (Balaenoptera physalis), humpback 

whale (Megaptera novaengliae), and right whale (Balaena glacialis). The plant species are: 

seabeach amaranth (Amaranthus pumilus), Canby's dropwort (Oxypolis canbyi), pondberry

(Lindera melissifolia) and American chaffseed (Schwalbea americana).

The shortnose Sturgeon, West Indian manatee, Kemp's ridley sea turtle, leatherback sea turtle,

green sea turtle, loggerhead sea turtle, West Indian manatee, finback whale, humpback whale, and 

right whale are all aquatic animals and would not be affected by the field test. The birds (piping 

plover, red-cockaded woodpecker and wood stork) could all potentially visit the field test site but 

would not nest there. The piping plover nest along the coast. The red-cockaded woodpecker nests 

in old growth longleaf pine. The wood stork nests in marshes, floodplain lakes, and swamps. The 

frosted flatwoods salamander inhabits longleaf or slash pine forests lying between drier land 
Page 78 of 98

upslope and wetlands and seasonally inhabits wet pine flat-woods with vernal pools. Discussions 

with Laura Zimmerman (contacted February 14 and 20, 2008) of the Charleston Field Office of the 

USFWS indicate that the species is not known to be in the area of the release. Known populations 

in the county are far from the release site in the Santee Coastal Reserve. A check of the SC 

Heritage Trust Database did not identify any occurrences in the area of the release. 

For the plants, the seabeach amaranth occurs in areas just above the high tide line on accreting 

shorelines; those where the beach is building up or expanding. Canby's dropwort occurs in a 

variety of coastal plain communities, including pond cypress savannahs, the shallows and edges of 

cypress and pond pine ponds, sloughs, and wet pine savannas. Pondberry is found in swamp and 

pond margins, sandy sinks, swampy depressions or wet flats that are subject to drying but the roots 

are submerged at times. American chaffseed is found in various sandy soil areas on the coastal 

plain; plants are usually found on margins of savannas and cypress ponds that are seasonally wet; 

best managed by prescribed fire. The location of the field test, which has been a managed forest 

plantation for more than 10 years, would not be hospitable habitat for any of these species.

Critical Habitat: There is designated critical habitat for the piping plover in Charleston County but 

it is along the coast and over 15 miles from the field test. Notice of designation of critical habitat 

for the frosted flatwood salamander (final rule) was published in the Federal Register on February 

10, 2009 (http://edocket.access.gpo.gov/2009/pdf/E9-2403.pdf). Unit FFS-7 is within Charleston 

County. This is located in the Santee Coastal Reserve which is over 50 miles away from the 

Charleston County site. 

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not adversely modify designated critical habitat or habitat proposed for 

designation.

Marlboro County, SC (SC-MAR-01)

This location has been a commercial nursery for forest seedling production for over 30 years. Site 

preparation involved herbicide application, subsoiling and planting of trees in flat beds. A 0.3 acre 

field test of some of the lines included in these permit requests were established in 2007, now 

covered under permit 08-039-102rm but subsequently terminated. The surrounding areas consist of 

field plantings of agricultural crops, nursery beds of forest tree seedlings and less than 30 year-old 

mixed hardwood and pine plantations. No additional trials are currently planned for this site.

There are two TES animals listed for Marlboro County; the red-cockaded woodpecker (Picoides 

borealis) and the shortnose sturgeon (Acipenser brevirostrum). The listed plant is Canby's 

Dropwort (Oxypolis canbyi). 
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The red-cockaded woodpecker nests in old growth longleaf pine not found at the location of the 

field test. The shortnose sturgeon occurs in rivers and estuaries. Little is known about the status 

of any of the populations in South Carolina. The closest body of water where the sturgeon could 

possibly live is about 5 miles away. Canby's dropwort occurs in a variety of coastal plain 

communities, including pond cypress savannahs, the shallows and edges of cypress and pond pine 

ponds, sloughs, and wet pine savannas. The location of the field test, which has been a managed 

nursery for more than 30 years, would not be hospitable habitat for any of these species.

Critical Habitat: There is no designated critical habitat or habitat proposed for designation in the

county.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Hardin County, TX (TX-HAR-01)

This location has been a managed forest plantation for more than 30 years. The location consists of 

mixed hardwood tree plantations planted using standard silvicultural practices and was harvested 

by the owner in 2004. The test site is within the larger harvested area and was re-bedded by the 

owner for planting. Site preparation included herbicide application and sub-soiling. A field test of 

19.7 acres of some of the lines included in these permit requests were established under permit 08-

039-102rm in 2009. The areas surrounding the test site consist of mixed hardwood stands and 

managed loblolly pine plantations.

One TES animal and one TES plant are listed for Hardin County. The animal is the red-cockaded 

woodpecker (Picoides borealis), and the plant is the Texas trailing phlox (Phlox nivalis ssp. 

texensis). 

The red-cockaded woodpecker might visit the field test site but would not nest there since it prefers 

old growth pine forests, particularly longleaf pine. The Texas trailing phlox is endemic to the 

Pineywoods of the west gulf coastal plain of east Texas. It occurs in deep sandy soils in firemaintained openings in upland longleaf pine savannas or post oak-bluejack oak woodlands. Since 

this site has been under managed forest plantations for more than 30 years this species would not 

find the field test site a suitable habitat.

Critical Habitat: There is no designated critical habitat or proposed critical habitat listed for this 

county.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 
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species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Jasper County, TX (TX-JAS-01 and TX-JAS-02)

There are two locations in Jasper County Texas. The first site, TX-JAS-01, has been a managed 

pine plantation for more than 25 years. Previous plantings were cultivated in beds using standard 

silvicultural practices. The existing pine plantation at this site was harvested by the owner in 2007 

and re-bedded for planting. The test site is within the larger harvested and bedded site prepared by 

the site owner. Further site preparation involved herbicide application, subsoiling and planting of 

trees in flat beds. A field test of 0.7 acre of some of the lines included in these permit requests 

were established under permit 08-039-102rm in 2008. The areas surrounding the test site consist of 

harvest age pine plantations.

The second site, TX-JAS-02, has been in agriculture for more than 30 years. The test site was used 

for pasture for the past 10 years. Site preparation involved herbicide application to remove existing 

grasses, subsoiling, preparation for irrigation installation, and flat plantings of trees. A field test of 

1.0 acre of some of the lines included in these permit requests were established under permit 08-

039-102rm in 2008. The areas surrounding the test site consist of natural stands of mixed pine and 

hardwoods.

Two threatened or endangered species (TES) animals and one plant are listed in Jasper County. 

The animals are the red-cockaded woodpecker (Picoides borealis) and the Louisiana black bear 

(Ursus americanus luteolus). The plant is Navasota ladies'-tresses (Spiranthes parksii).

The red-cockaded woodpecker (Picoides borealis) inhabits old growth forests, primarily longleaf 

pine and might visit the field test sites but would not nest there. The Louisiana black bear (Ursus 

americanus luteolus) depends on diverse, productive bottomland forest with diverse food resources, 

including a variety of hard-mast-producing species. High quality habitat includes remote areas 

with little or no human activity so it would not likely occur at either site. Navasota ladies'-tresses 

(Spiranthes parksii) occurs at the margins of post oak (Quercus stellata) woodlands in sandy loams 

along intermittent tributaries of rivers. The field test sites have none of these characteristics.

Critical Habitat: There is no designated critical habitat or habitat proposed for designation in the

county.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at these release sites. If they were to enter the sites, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The sites are not within or near designated critical habitat or habitat proposed 

for designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.
Page 81 of 98

Jefferson County, TX (TA-JEF-01)

This location has been used for managed agricultural production of rice for more than 5 years. Site 

preparation will involve herbicide application, subsoiling and planting of trees in flat beds. The 

surrounding areas of the test site consist of rice plantations.

There are six TES animals listed for Jefferson County. There are no plants listed. The animals are

green turtle (Chelonia mydas), hawksbill turtle (Eretmochelys imbricata imbricate), Kemp's ridley 

sea turtle (Lepidochelys kempii), leatherback turtle (Dermochelys coriacea), loggerhead sea turtle

(Caretta caretta) and piping plover (Charadrius melodus). 

All of the turtles occur in the open waters of the Gulf of Mexico or in shallow coastal and estuarine 

waters. The field test site is about 30 miles away from the Gulf. The piping plover uses sparsely 

vegetated dunes and coastal beaches in southern Jefferson County, which is also at least 30 miles 

from the field test site. None of these species would be impacted by the field test. 

Critical Habitat: There is critical habitat listed for the piping plover but it does not occur in this 

county. The closest critical habitat is in neighboring Galveston County, approximately 40 miles 

from the field test site.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Newton County, TX (TX-NEW-01, TX-NEW-02, TX-NEW-03, TX-NEW-04)

There are four locations in Newton County. All locations have been under managed loblolly pine 

plantations for at least 30 years. The previous plantings were cultivated in beds using standard 

silvicultural practices and the areas have recently been harvested. Site preparation has been or will 

include herbicide application, plowing, and planting of trees in raised or flat beds. The surrounding 

areas of the test site consist of managed loblolly pine plantations and/or mixed hardwood stands.

Two TES animals are listed for Newton County. The animals are the red-cockaded woodpecker 

(Picoides borealis), and Louisiana black bear (Ursus americanus luteolus). There are no plants 

listed for this county.

The red-cockaded woodpecker might visit the field test site but would not nest there since it prefers 

old growth pine forests, particularly longleaf pine. The Louisiana black bear (Ursus a. luteolus) 

prefers bottomland forests with diverse food resources, including a variety of hard-mast-producing 

species. According to the Black Bear Recovery Plan 

(http://www.bbcc.org/web/images/stories/information/pdf/FinalRestorationPlanwithFigures.pdf) in 
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east Texas, potentially occupied habitat may occur in at least four Counties: Cass, Shelby, Panola 

and Angelina Counties. All of these locations are from fifty to over a hundred miles away from 

these test sites. Louisiana black bear habitat includes remote areas with little or no human activity 

so it is unlikely to be found at the site.

Critical Habitat: The proposed critical habitat is located in Avoyelles, East Carroll, Catahoula, 

Concordia, Franklin, Iberia, Iberville, Madison, Pointe Coupee, Richland, St. Martin, St. Mary, 

Tensas, West Carroll, and West Feliciana Parishes, Louisiana. All of these locations are over a 

hundred miles away from these test sites. 

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at these release sites. If they were to enter the sites, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The sites are not within or near designated critical habitat or habitat proposed 

for designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Bay County, FL (FL-BAY-01)

This location was used as an intensely managed pasture for more than 15 years, and was planted 

with grasses suitable for cattle grazing. Site preparation involved herbicide application to remove 

existing grasses, subsoiling, preparation for possible irrigation installation, and planting of trees in 

flat beds. Field tests of some of the lines included in these permit requests were established in 2007 

(1.0 acre) and 2008 (0.35 acres), now covered under permit 08-039-102rm. All tests except 0.18 

acre were subsequently terminated. The areas surrounding the test site consist of experimental 

forest trees, agricultural crops and less than 25 year-old hardwoods and pine.

Sixteen TES animals and 6 TES plants are listed for Bay County. The animals are gulf 

moccasinshell (Medionidus penicillatus), oval pigtoe (Pleurobema pyriforme), gulf sturgeon 

(Acipenser oxyrinchus) loggerhead turtle (Caretta caretta), eastern indigo snake (Drymarchon 

corais couperi), piping plover (Charadrius melodus), red-cockaded woodpecker (Picoides 

borealis), West Indian manatee (Trichechus manatus), Choctawhatchee beach mouse (Peromyscus 

polionotus allophrys), St. Andrews beach mouse (Peromyscus polionotus peninsularis), reticulated 

flatwoods salamander (Ambystoma bishopi), green turtle (Chelonia mydas), leatherback turtle 

(Dermochelys coriacea), hawksbill turtle (Eretmochelys imbricata imbricate), Kemp's ridley turtle 

(Lepidochelys kempii), and wood stork, (Mycteria americana). The plants are telephus spurge 

(Euphorbia telephioides), white birds-in-a-nest (Macbridea alba), Godfrey’s (violet) butterwort 

(Pinguicula ionantha), Florida skullcap (Scutellaria floridana), Harper's beauty, (Harperocallis 

flava) and Crystal Lake nailwort, (Paronychia chartacea minima).

The gulf moccasinshell, oval pigtoe, gulf sturgeon, West Indian manatee, and the turtles 

(loggerhead, green, leatherback, hawksbill and Kemp's ridley) are all aquatic animals and would 

not be affected by the field test. The birds (piping plover, red-cockaded woodpecker and wood 

stork) could all potentially visit the field test site but would not nest there. The piping plover nests

along the coast which is about 20 miles away from the release site. The red-cockaded woodpecker 
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nests in old growth longleaf pine. The wood stork nests in marshes, floodplain lakes, and swamps. 

The two mouse species live along the coast in beach areas so would not be impacted by the field 

release. For the eastern indigo snake, according to Caroline Stahaller (contacted February 20, 

2008) of the Panama City Field Office USFWS, the species has not been positively identified in the 

county or anywhere else on the Florida panhandle in 15-20 years. Considering the area is currently 

managed as a pasture, a habitat where the species is unlikely to occur, there is no concern for the 

species at this site. Although it is highly unlikely that the species would be found at the site, the 

applicant will provide all workers with identifying characteristics of the snake and instructions on 

what to do if the species is encountered. These measures are a variation of standard protective 

measures the USFWS uses when they have reached a “may affect” determination for construction 

sites. The reticulated flatwoods salamander inhabits longleaf or slash pine forests lying between 

drier land upslope and wetlands and seasonally inhabits wet pine flat-woods with ephemeral 

wetlands. They typically breed in the low wetlands where they lay their eggs. After hatching, they 

spend 11 to 18 weeks as larvae before metamorphosing into adults and leaving the wetland for 

higher ground to burrow into the soil. It is important that the area through which the species moves 

is vegetated with grasses. Management of the area with controlled burning is generally needed to 

provide quality flatwoods salamander habitat. Reticulated flatwoods salamanders may move as far 

as 450 meters from their breeding sites. Although the release site is not an area with known 

populations, and is at least 10 miles from the critical habitat, Ms. Stahaller of USFWS suggested a 

closer look at the site to determine if the habitat would be suitable for the species. The property 

owner was forwarded survey protocol information provided by USFWS. In late March 2008, the 

owner, (who is 64 and has lived at the site his entire life) walked the site within 450 meters of the 

release. The owner was contacted on April 7, 2008 to discuss his observations. The owner looked 

for adult salamanders but did not find any, and does not recall ever seeing them at the site. There 

were four naturally wet areas that were found. One was in the pasture itself. It was a small site of 

about 8 ft. by 10 ft. and is surrounded by an area that has been in continuous agricultural use for at 

least 60 years. The other two sites were heavily wooded areas surrounded by pasture and remain 

constantly wet in most years. They are approximately 250 yards from the release site. The fourth 

site is approximately 400 to 500 yards from the release site and is a 5 acre wetland that shares the 

same elevation with an adjacent creek and remains constantly wet, but does dry around the 

perimeter. Vegetation in the site is longleaf pine with a wiregrass groundcover – habitat the species 

finds most suitable. Vegetation between this potential breeding site and the release site is pasture. 

It is unlikely that the flatwoods salamander would be using the release site for a number of reasons. 

First, the only possible breeding sites within 450 meters of the release remain constantly wet during 

most years, a condition that would not serve as breeding habitat. The five acre site where the 

perimeter dries could perhaps allow for breeding in some years but it is about at the farthest 

possible distance from the release site that the adults are known to travel from their breeding area. 

Second, the sites are either surrounded by land that has been used for agricultural purposes for at 

least 60 years or is separated from the release site by such land. Activities associated with 

agricultural production, disking, plowing, application of pesticides etc. would make the habitat 

unsuitable for the species. In addition, none of this area is managed by controlled burning, a 

management practice that is generally used in areas where the species has viable populations. The 

lack of suitable breeding habitat, combined with the ongoing and historic agricultural activities at 

the site, point to a conclusion that the reticulated flatwood salamander will not be present in the 

area of the release, nor will they be likely to enter the site during the field trial. None of the six

listed plant species would be expected to grow in the existing pasture. They grow in wiregrass 
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dominated longleaf pine savannahs, in the mesic flatwoods, on low sand ridges of pine-scrub oak 

near the Gulf of Mexico or in wet prairies and seepage bogs.

This location is being converted from a pasture to a forest tree research plot. Based on the above 

evaluation, APHIS has determined that disturbance of the field site for the release (e.g. plowing, 

removing vegetation, burning etc.) will not directly or indirectly affect a federally listed threatened 

or endangered species or species proposed for listing, or affect the habitat for these species.

Critical Habitat: There is designated critical habitat for the Gulf sturgeon along the entire coast of 

Bay County. Critical habitat for the Choctawhatchee beach mouse, St. Andrew’s beach mouse, and 

the piping plover is along the coast in the Panama City area. The field test site is approximately 18 

miles away from the coast so would not impact these critical habitats. Notice of designation of 

critical habitat for the reticulated flatwood salamander (final rule) was published in the Federal 

Register on February 10, 2009 (http://edocket.access.gpo.gov/2009/pdf/E9-2403.pdf). Unit RFS-6, 

subunit B is just beyond the northern border of Bay County in neighboring Washington County. 

The field test site is approximately 28 miles away from this location. Unit RFS-9 subunit A is 

located in neighboring Calhoun County approximately 10 miles from the site. There is proposed 

critical habitat listed for the West Indian manatee but it does not occur in this county.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Columbia County, FL (FL-COL-01)

This location has been under managed pine plantations for more than 20 years. Previous plantings 

were cultivated in beds using standard silvicultural practices. This area of the tract was burned in a 

fire in May 2007. After the fire, the area was raked and bedded by the site owner in preparation for

re-planting. The test site is within the larger harvested and bedded area, which is surrounded by 

existing pine plantations and additional harvested tracts.

Five TES animals are listed for Columbia County. The animals are oval pigtoe (Pleurobema 

pyriforme), gulf sturgeon (Acipenser oxyrinchus), eastern indigo snake (Drymarchon corais 

couperi), red-cockaded woodpecker (Picoides borealis), and wood stork (Mycteria americana). 

There are no plants listed for this county.

The oval pigtoe and gulf sturgeon are aquatic animals and would not be affected by the field test. 

The birds (red-cockaded woodpecker and wood stork) could all potentially visit the field test site 

but would not nest there. The red-cockaded woodpecker nests in old growth longleaf pine. The 

wood stork nests in marshes, floodplain lakes, and swamps. For the eastern indigo snake, 

according to Caroline Stahaller (contacted February 20, 2008) of the Panama City Field Office 

USFWS, the species has not been positively identified in the Florida panhandle in 15-20 years. 
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Considering the area has been managed as a pine plantation for many years, it is unlikely to be 

found at this site. Although it is highly unlikely that the species would be found at the site, the 

applicant will provide all workers with identifying characteristics of the snake and instructions on 

what to do if the species is encountered. These measures are a variation of standard protective 

measures the USFWS uses when they have reached a “may affect” determination for construction 

sites.

Critical Habitat: There is designated critical habitat for the Gulf sturgeon but it is not listed in this 

county. There is no proposed critical habitat listed for this county.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Gadsden County, FL (FL-GAD-01, FL-GAD-02)

There are two planting sites at this location. One location, FL-GAD-01, has been an agricultural 

research station for more than 10 years and has been used for conducting research experiments on 

agricultural crops. Standard silvicultural practices were used for site preparation, including 

herbicide application, plowing and planting of trees in raised or flat beds. A field test of 0.2 acre of 

some of the lines included in these permit requests were established under permit 08-039-102rm in 

2008. The areas surrounding the test site consist of mixed pine-hardwood forests and pine 

plantations as well as research plantings of agricultural and horticultural crops.

The second location, FL-GAD-02, has been an agricultural research station for more than 10 years. 

The field has been fallow for approximately seven years. Standard silvicultural practices will be 

used for site preparation, including herbicide application, plowing and planting of trees in raised or 

flat beds. The areas surrounding the test site consist of mixed pine-hardwood forests and pine 

plantations, as well as research plantings of agricultural and horticultural crops.

Ten TES animals and six TES plants are listed for Gadsden County. The animals are: 

fat threeridge mussel (Amblema neislerii), purple bankclimber mussel (Elliptoideus sloatianus), 

shinyrayed pocketbook (Hamiota subangulata), ochlockonee moccasinshell (Medionidus 

simpsonianus), oval pigtoe (Pleurobema pyriforme), chipola slabshell (Elliptio chipolaensis), gulf 

sturgeon (Acipenser oxyrinchus desotoi), wood stork (Mycteria americana), red-cockaded 

woodpecker, (Picoides borealis), Eastern indigo snake (Drymarchon corais couperi). The plants 

are: Florida torreya (Torreya taxifolia), Chapman's rhododendron (Rhododendron chapmanii), 

American chaffseed (Schwalbea americana), fringed campion (Silene polypetala), Miccosukee 

gooseberry (Ribes echinellum) and gentian pinkroot (Spigelia gentianoides).

The six mussel species (fat threeridge mussel, purple bankclimber mussel, shinyrayed pocketbook,

Ochlockonee moccasinshell, oval pigtoe and chipola slabshell), and the gulf sturgeon (Acipenser 
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oxyrinchus desotoi) are all aquatic animals and would not be affected by the field test. (See CH 

info below on the gulf sturgeon). The red cockaded woodpecker (Picoides borealis) inhabits old 

growth forests, primarily longleaf pine. It could visit the field test site but would not nest there. 

The wood stork (Mycteria americana) primarily inhabits wetland systems notably cypress or 

mangrove swamps and would not find the field test site hospitable. For the Eastern indigo snake 

(Drymarchon corais couperi), according to Caroline Stahaller, (contacted February 20, 2008) of the 

Panama City Field Office USFWS, the species has not been seen anywhere on the Florida 

panhandle in 15-20 years. She had no concern that the action could affect the species and agreed 

the action would have “no effect.” Although it is highly unlikely that the species would be found 

at the site, the applicant will provide all workers with identifying characteristics of the snake and 

instructions on what to do if the species is encountered. These measures are a variation of standard 

protective measures the USFWS uses when they have reached a “may affect” determination for 

construction sites.

None of the six listed plant species would be expected to grow at the field site/research station. For 

American chaffseed (Schwalbea Americana), according to the species recovery plan, the only 

known extant population in the county is on private land. A site visit in 1994 found the area 

changed to a residential development. It is assumed that the population may be extirpated. For 

Chapman’s rhododendron (Rhododendron chapanii), according to the Federal Register notice 

listing the species, there is only one population in Gadsden County. The population is on land 

owned by a paper company near the Gadsden – Liberty county line, and is many miles from the 

release site. The Florida torreya (Torreya taxifolia) occurs on hardwood hammock slopes, ravines, 

and bluffs of the Apalachicola River region, usually in steephead ravines (deep cuts made by 

erosion into coastal plain sediments). The ravines are much cooler and more moist than the land 

surface above and harbor remnants of the more temperate flora that existed in the region during the 

Tertiary ice ages. The fringed campion (Silene polypetala) prefers well-drained, sandy-loam soils 

of deciduous woods, usually hillsides. The Miccosukee gooseberry (Ribes echinellum) is only 

known to exist on sites along east bank of Lake Miccosukee which is about 36 miles away from the 

field test site. The gentian pinkroot (Spigelia gentianoides) is also listed in Gadsden County and 

specimens were once collected in the county, but presently the only population currently known to 

exist is at the Three Rivers State Recreation Area, Lake Seminole, Jackson County, FL. 

(http://www.centerforplantconservation.org/). None of these species would find the field test site 

as suitable habitat.

Critical Habitat: The designated critical habitat for the gulf sturgeon includes the Apalachicola

River mainstream, beginning from the Jim Woodruff Lock and Dam, Gadsden and Jackson 

Counties, Florida, downstream to its discharge at East Bay or Apalachicola Bay, Franklin County, 

Florida. The establishment of the field test site would not impact this habitat. The field site is 

about 19 miles away from the Apalachicola river mainstream below the Jim Woodruff Lock and 

Dam. The mussel species have designated critical habitat, but the nearest to the release site is in the 

Ochlokonee River over 10 miles away.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to found at these release sites. If they were to enter the sites, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 
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proposed for listing. The sites are not within or near designated critical habitat or habitat proposed 

for designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Highlands County, FL (FL-HIG-01)

This location was previously used for managed production of citrus for at least 15 years. The 

planting area at this location had been used for field trials of transgenic Eucalyptus for more than 6 

years. Site preparation involved herbicide application, plowing, and planting of trees in flat beds. 

Field trials of 1.4 acres of some of the lines included in these permit requests are being grown 

under issued permit # 08-151-101r under which these trees are allowed to flower. The oldest of 

these trees at this site are now entering their fifth growing season. An additional 2.3 acres of tests 

of different lines with the same EH1 hybrid are being grown under permits 08-039-102rm and 09-

070-101rm. Areas surrounding the test site consist of less than 5 year-old second-growth pine and 

hardwood with mixed grasses.

Fifteen TES animals, nineteen TES plants, and one TES lichen are listed for Highlands County. 

The animals are: American alligator (Alligator mississippiensis), wood stork (Mycteria americana), 

Audubon’s crested caracara (Caracara cheriway), red-cockaded woodpecker (Picoides borealis), 

Florida scrub-jay (Aphelocoma coerulescens), Florida grasshopper sparrow (Ammodramus 

savannarum floridanus), Everglade snail kite (Rostrhamus sociabilis plumbeus), ivory-billed 

woodpecker (Campephilus principalis), Whooping crane (Grus americana) (non-essential 

experimental population), Florida panther (Puma concolor coryi), Puma (Puma concolor –

similarity of appearance), West Indian manatee (Trichechus manatus), Eastern indigo snake 

(Drymarchon corais couperi), bluetail mole skink (Eumeces egregius lividus) and sand skink 

(Neoseps reynoldsi).

The plants are: Florida bonamia (Bonamia grandiflora), pygmy fringetree (Chionanthus 

pygmaeus), pigeon wings (Clitoria fragrans), short-leaved rosemary (Conradina brevifolia), Avon 

Park hare-bells (Crotalaria avonensis), Garrett’s mint (Dicerandra christmanii), scrub mint 

(Dicerandra frutescens), scrub buckwheat (Eriogonum longifolium var. gnaphalifolium), snakeroot 

(Eryngium cuneifolium), highlands scrub hypericum (Hypericum cumulicola), scrub blazingstar 

(Liatris ohlingerae), Britton's bear-grass (Nolina brittoniana), papery whitlow-wort (Paronychia 

chartacea ssp. Chartacea), Lewton's polygala (Polygala lewtonii), wireweed (Polygonella 

basiramia), sandlace (Polygonella myriophylla), scrub plum (Prunus geniculata), Carter's mustard 

(Warea carteri), and Florida ziziphus (Ziziphus celata).

The lichen is: Florida perforate cladonia (Cladonia perforate).

The American alligator (Alligator mississippiensis) is no longer biologically endangered or 

threatened; however, it is listed by USFWS as Threatened throughout its entire range due to 

similarity of appearance to other endangered or threatened crocodilians. The wood stork (Mycteria 

Americana) primarily inhabits wetland systems notably cypress or mangrove swamps and would 

not find the field test site hospitable. The crested caracara (Caracara cheriway) is associated with 

open country; dry prairie with scattered cabbage palms, wetter prairies, and to some extent also 

improved pastures and sometimes wooded areas having associated limited areas of open grassland. 
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The center of range is the Kissimmee Prairie, an area of shallow ponds and sloughs with scattered 

hummocks of live oaks and cabbage palms. The red cockaded woodpecker (Picoides borealis) 

inhabits old growth forests, primarily longleaf pine. The Florida scrub-jay (Aphelocoma 

coerulescens) prefers oak scrub on white, drained sand, in open areas without a dense canopy 

associated with Palmetto, sand pine and rosemary. This includes scrub with no canopy, sandpine 

scrub, scrubby flatwoods, and coastal scrub. The Florida grasshopper sparrow (Ammodramus 

savannarum floridanus) prefers dry prairie with stunted saw palmetto and dwarf oaks, bluestems 

and wiregrass and unimproved cattle pastures. Its habitat is maintained by periodic fires. The snail 

kite (Rostrhamus sociabilis plumbeus) prefers large, open freshwater marshes and lakes with 

shallow open waters. The ivory-billed woodpecker (Campephilus principalis) formerly occurred in 

the southeastern United States and Cuba and has declined to extinction or near extinction. It once 

occupied swampy forests, especially large bottomland river swamps of coastal plain and 

Mississippi Delta and cypress swamps of Florida, in areas with many dead and dying trees. It 

would not occur in an agricultural environment such as the field test site. Whooping crane (Grus 

americana) prefers freshwater marshes and wet prairies. It nests in dense emergent vegetation 

(sedge, bulrush) in shallow ponds, freshwater marshes, wet prairies, or along lake margins, within 

large expanses of undisturbed wilderness. The cranes listed in this county are an experimental 

reintroduction of whooping cranes in Florida initiated in 1993 to establish a non-migratory 

population. This is an experimental, non-essential population. Any of the bird species could 

potentially visit the field test site but would not nest there. The Florida panther (Puma concolor

coryi) generally occurs in heavily forested areas in lowlands and swamps, also upland forests in 

some parts of range; areas with adequate deer or wild hog population. Habitats include tropical 

hammocks, pine flatwoods, cabbage palm forests, mixed swamp, cypress swamp, live oak 

hammocks, sawgrass marshes, and Brazilian pepper thickets. It depends on large contiguous 

blocks of wooded habitat, though interspersed fields and early successional habitats may be 

beneficial through their positive effect on prey populations. Its day-use sites typically are dense 

patches of saw palmetto surrounded by swamp, pine flatwoods, or hammock. It would not occur in 

the trial area due to the openness and continued presence of humans in the area. The West Indian 

manatee (Trichechus manatus) occurs in shallow coastal waters, rivers, bays and lakes; none of 

which are close to any of this release location. For the Eastern indigo snake (Drymarchon cosair 

couperi) – according to Candice Martino (904-232-2580 ext. 129) Section 7 Endangered Species 

biologist from the Jacksonville, FL Field Office, (contacted February 25, 2008) the species is 

seldom seen but could be anywhere. However, the habitat at the release site would not be suitable. 

The historic and continuous use of the release site and the surrounding area as a citrus grove and 

Eucalyptus field trials makes it extremely unlikely that the species would be found in the area. 

Therefore, the appropriate determination would be “no effect.” Although it is highly unlikely that 

the species would be found at the site, the applicant will provide all workers with identifying 

characteristics of the snake and instructions on what to do if the species is encountered. These 

measures are a variation of standard protective measures the USFWS uses when they have reached 

a “may affect” determination for construction sites.

The bluetail mole skink (Eumeces egregius lividus) inhabits sand pine-rosemary scrub or, less 

frequently, longleaf pine-turkey oak association sandhills. It occupies localized pockets of 

sufficient leaf litter and moisture to provide abundant food and nesting sites. The sand skink 

(Neoseps reynoldsi) occurs only on Florida's central ridges, at elevations of 27 m or more. It 

inhabits loose sands of sand pine-rosemary scrub, less often longleaf pine-turkey oak sandhills or 
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turkey oak barrens adjacent to scrub, especially high pine-scrub ecotones. It was determined that 

the release site is within a geographic area where these two skink species are found. According to 

Brad Rick, (contacted February 27 and 28, 2008) of the Vero Beach Field Office USFWS, sand 

skinks and bluetail mole skinks are found in scrub habitat with areas of open sand. The literature 

indicates that skinks are sometimes found in active and abandoned citrus groves and the applicant 

confirmed that sandy soils are predominant in the area. It was decided to have the applicant 

conduct a survey of the species using USFWS protocols to determine if the species is present. The 

protocols were provided by the Vero Beach Field Office. The USFWS protocol recommends that 

surveys be conducted between March 1 and May 15 as this is an ideal time to observe evidence of 

the skinks. A coverboard survey was conducted over a one month period from March 18 to April 

15. The coverboards were checked weekly on March 25, April 1, April 8 and April 15. No 

evidence indicating the presence of sand skinks or bluetail mole skinks was observed and they are 

presumed absent. Therefore, the appropriate determination would be “no effect.”

For the plants, the Florida bonamia (Bonamia grandiflora) grows in natural clearings of bare 

ground and invades disturbed areas of open sand. Although not common, it is often locally 

abundant where there is little or no shade from trees or shrubs. It is locally abundant on deep, 

white, dry sands of ancient dunes and sandy ridges in clearings or openings of scrub habitat on the 

Central Ridge of Florida. Pygmy fringetree (Chionanthus pygmaeus) is generally found in xeric, 

coarse white sand of scrub/oak scrub areas found at the southern end of the Central Florida Ridge. 

It is also found occasionally in longleaf pine-turkey oak vegetation, high pineland, dry hammocks, 

and transitional habitats. Pigeon wings (Clitoria fragrans) is widely scattered in undisturbed 

clearings of xeric sandhill and scrub communities on well-drained upland soils. It is typically 

found in undisturbed clearings in scrub areas but also occurs in very open scrub as well. Shortleaved rosemary (Conradina brevifolia), is found in white sands of sand pine-oak scrub with 

scattered overstory of sand pine and scrub oak. Avon Park hare-bells (Crotalaria avonensis) 

occurs in upland habitats (scrub and sandhill), often along trails. It grows in full sun or partial 

shade provided by characteristic scrub shrubs or sand pine. Garrett’s mint (Dicerandra 

christmanii) occurs in openings in sand pine-oak scrub on yellow soils of the Central Florida Ridge. 

Scrub mint (Dicerandra frutescens) occurs in well-drained soils of scrub or sandhill vegetation. It 

is locally abundant in and around the sand pine-evergreen oak scrub, where it may occur in the low 

shrub layer or in open stands, clearings, or adjacent sandy places. It is not found in areas cleared 

for pasture, or areas in which wholesale site preparation has taken place. Scrub buckwheat 

(Eriogonum longifolium var. gnaphalifolium) is long-lived, slow growing and flowers and 

reproduces primarily after fires or other disturbances (e.g. logging, mowing) that increase light 

availability. It prefers dry pinelands, sandhills, and scrub (longleaf pine-turkey oak, scrub oaks) 

and is more commonly found in transition habitats between scrub and high pine and in turkey oak 

barrens than in either dense scrub or open high pine. Snakeroot (Eryngium cuneifolium) is 

generally found in areas of open sand, including blowouts and other highly disturbed soil surfaces, 

such as road shoulders. It occurs in exposed sunny openings; areas in scrub, especially rosemary 

scrub. Highlands scrub hypericum (Hypericum cumulicola) occurs in patches of open, nutrientpoor sand within oak and rosemary scrub. It is often associated with reindeer lichen (Cladonia

spp.) and snakeroot (Eryngium cuneifolium). Scrub blazingstar (Liatris ohlingerae) occurs in 

openings in oak-rosemary scrub and sand pine scrub. Britton's bear-grass (Nolina brittoniana) 

occurs in deep, fine-textured, well-drained sands of sand pine-evergreen oak scrub or longleaf pineturkey oak sandhills. Nolina is entirely dependent on fire or some other mechanism to maintain an 
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open successional stage in scrub or sandhills. Papery whitlow-wort (Paronychia chartacea ssp. 

Chartacea) is a sand scrub that occurs on ancient dunes in the lake region, in white sand clearings 

or blowouts. Lewton's polygala (Polygala lewtonii) occurs in sandhills characterized by longleaf 

pine and low scrub oaks, including low turkey oak woods, and transitional sandhill/scrub habitats. 

This species occasionally inhabits powerline clearings or new roadsides. Wireweed (Polygonella 

basiramia) is restricted to bare patches within sand pine-evergreen oak scrub vegetation. It grows 

on areas of bare sand within sand pine (Pinus clausa) and Florida rosemary (Ceratiola ericoides). 

Sandlace (Polygonella myriophylla) occurs in areas of sand pine scrub and ancient sand dunes. 

Scrub plum (Prunus geniculata) has a very narrow range and small widely scattered populations. It 

frequently forms small colonies of several plants but may grow as solitary individuals. It grows in 

deep, yellow sands of longleaf pine-turkey oak sandhill and white, excessively leached, winddeposited soils of evergreen scrub oak-sand pine scrub. Carter's mustard (Warea carteri) occurs in 

sandy clearings in sand scrub and sandhills; scattered overstory of sand; longleaf or slash pine and 

scrub oaks. Florida ziziphus (Ziziphus celata) is a scrub that occurs on gently rolling hills with 

vegetation dominated by Carya floridana and Quercus species. It prefers open, sunny areas. The 

Perforate Reindeer Lichen (Cladonia perforate) occurs in sandy openings in stabilized sand dunes 

with Florida scrub vegetation. It is often associated with Ceratiola. None of the plants and the 

lichen listed above would find the field test site as suitable habitat and would not be present given 

the historic and continuous use of the release site and the surrounding area as a citrus grove and 

research area used for growing Eucalyptus.

Critical Habitat: There is proposed critical habitat listed for the West Indian manatee but none of 

the proposed habitat occurs in this county. The whooping crane population in this county is a nonessential experimental population and does not have critical habitat.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at the release site. If they were to enter the site, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Marion County, FL (FL-MAR-01)

This location has been used as an agricultural research station for more than 5 years for conducting 

research experiments on agricultural crops. Site preparation involved herbicide application, 

plowing, and planting of trees in flat beds. A field test of 0.9 acre of some of the lines included in 

these permit requests were established under permit 08-039-102rm in 2008. In addition, a field test 

of 0.3 acres of different lines with the same EH1 hybrid was planted in 2009 under permit 9-070-

101rm. Areas surrounding the test site consist of agricultural fields and plantings of horticultural 

crops. 

Seven TES animals and four TES plants are listed for Marion County. The animals are: sand skink 

(Neoseps reynoldsi), Eastern indigo snake (Drymarchon corais couperi), wood stork (Mycteria

americana), Everglades snail kite (Rostrhamus sociabilis plumbeus), red-cockaded woodpecker 
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(Picoides borealis), Florida scrub-jay (Aphelocoma coerulescens), and West Indian manatee 

(Trichechus manatus). The plants are: Florida bonamia (Bonamia grandiflora), longspurred mint 

(Dicerandra cornutissima), scrub buckwheat (Eriogonum longifolium var. gnaphalifolium), and 

Lewton's polygala (Polygala lewtonii).

The Everglades snail kite (Rostrhamus sociabilis plumbeus) prefers large, open freshwater marshes 

and lakes with shallow open waters. The red cockaded woodpecker (Picoides borealis) inhabits 

old growth forests, primarily longleaf pine. The Florida scrub-jay (Aphelocoma coerulescens) 

prefers oak scrub on white, drained sand, in open areas without a dense canopy associated with 

Palmetto, sand pine and rosemary. This includes scrub with no canopy, sandpine scrub, scrubby 

flatwoods, and coastal scrub. The wood stork (Mycteria americana) primarily inhabits wetland 

systems notably cypress or mangrove swamps and would not find the field test site hospitable. Any 

of the four bird species could potentially visit the field test site but would not nest there. The West 

Indian manatee occurs in shallow coastal waters, rivers, bays and lakes; none of which are close to 

any of this release location. For the Eastern indigo snake (Drymarchon corais couperi) – according 

to Candice Martino, Section 7 Endangered Species biologist from the Jacksonville, FL Field Office, 

(contacted February 25, 2008) the species is seldom seen but could be anywhere. However, the 

habitat at the release site would not be suitable. The historic and continuous use of the release site 

and the surrounding area as an agricultural research station makes it extremely unlikely that the 

species would be found in the area. Therefore, the appropriate determination would be “no effect”. 

Although it is highly unlikely that the species would be found at the site, the applicant will provide

all workers with identifying characteristics of the snake and instructions on what to do if the species 

is encountered. These measures are a variation of standard protective measures the USFWS uses 

when they have reached a “may affect” determination for construction sites. Also for the sand 

skink (Neoseps renoldsi) – according to Candice Martino (contacted February 25, 2008), although 

the release site may be within a greater geographic area where the species may be found, the habitat 

of the release site would not be suitable for the species. The historic and continuous use of the 

release site and the surrounding area as an agricultural research station makes it extremely unlikely 

that the species would be found in the area. Therefore, the appropriate determination would be “no 

effect”.

For the plants, the Florida bonamia (Bonamia grandiflora) grows in natural clearings of bare 

ground and invades disturbed areas of open sand. Although not common, it is often locally 

abundant where there is little or no shade from trees or shrubs. It is locally abundant on deep, 

white, dry sands of ancient dunes and sandy ridges in clearings or openings of scrub habitat on the 

Central Ridge of Florida. The longspurred mint (Dicerandra cornutissima) is scattered in openings 

(natural or artificial) in longleaf pine-turkey oak scrub/sandhill or on low rises in slash pinepalmetto scrub. The scrub buckwheat (Eriogonum longifolium var. gnaphalifolium) is an 

herbaceous perennial that occurs in dry pinelands, sandhills, and scrub (longleaf pine-turkey oak, 

scrub oaks). It is more commonly found in transition habitats between scrub and high pine and in 

turkey oak barrens than in either dense scrub or open high pine. Lewton's polygala (Polygala 

lewtonii), occurs in sandhills characterized by longleaf pine and low scrub oaks, including low 

turkey oak woods, and transitional sandhill/scrub habitats. This species occasionally inhabits 

powerline clearings or new roadsides. None of the four plant species listed above would find an 

agricultural research area as suitable habitat and would not be present given the historic and 

continuous use of the release site and the surrounding area as an agricultural research station.
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Critical Habitat: There is no designated critical habitat or habitat proposed for designation in the

county. There is proposed critical habitat listed for the West Indian manatee but it is not in this 

county.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to found at the release site. If they were to enter the site, their presence would be fleeting 

as the habitat is either not suitable or does not contain constituent elements required by the species. 

Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The site is not within or near designated critical habitat or habitat proposed for 

designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Taylor County, FL (FL-TAY-01, FL-TAY-02 and FL-TAY-03)

There are three release site locations in Taylor County. 

These locations have been under managed pine plantations for over 20 years. The previous 

plantings were cultivated in beds using standard silvicultural practices. The existing pine 

plantations at these sites were harvested by the owner and prepared for re-planting. Additional site 

preparation involved herbicide application, plowing to remove stumps, and planting of trees in 

raised beds. Approximately 9.3 acres of field tests of some of the lines included in these permits 

requests were established across these sites under permit 08-039-102rm in 2008 and 2009. Areas 

surrounding these test sites consist of managed stands of pine plantations. 

There are ten TES animals listed for Taylor County. There are no listed plants. The animals are: 

Eastern indigo snake (Drymarchon corais couperi), wood stork (Mycteria americana), piping 

plover (Charadrius melodus), red-cockaded woodpecker (Picoides borealis), West Indian manatee 

(Trichechus manatus), Gulf sturgeon (Acipenser oxyrinchus desotoi), green turtle (Chelonia 

mydas), leatherback turtle (Dermochelys coriacea), Kemp's ridley sea turtle (Lepidochelys kempii)

and loggerhead sea turtle (Caretta caretta).

According to Caroline Stahaller, (contacted February 20, 2008) of the Panama City Field Office 

USFWS, the Eastern indigo snake (Drymarchon couperi) has not been seen anywhere on the 

Florida panhandle in 15-20 years. She had no concern that the action could affect the species. 

Although it is highly unlikely that the species would be found at the site, the applicant will provide 

all workers with identifying characteristics of the snake and instructions on what to do if the species 

is encountered. These measures are a variation of standard protective measures the USFWS uses 

when they have reached a “may affect” determination for construction sites. The birds (piping 

plover, red-cockaded woodpecker and wood stork) could all potentially visit the field test site but 

would not nest there. The piping plover nests along the coast which is seven or more miles away 

from the field test sites. The red-cockaded woodpecker nests in old growth longleaf pine. The 

wood stork nests in marshes, floodplain lakes, and swamps. The remaining species: West Indian 

manatee (Trichechus manatus), Gulf sturgeon (Acipenser oxyrinchus desotoi), green turtle

(Chelonia mydas), leatherback turtle (Dermochelys coriacea), Kemp's ridley sea turtle 

(Lepidochelys kempii) and loggerhead sea turtle (Caretta caretta) are all aquatic species and would 
Page 93 of 98

not occur anywhere close to the field test site. The closest body of water is the Econfina river 

which is about 2.5 miles away from one of the test sites.

Critical Habitat: There is designated critical habitat listed for the piping plover in Taylor County. 

It is located along the coast in the southwest part of the county. It is approximately 15, 27 and 32 

miles away from the three test sites. There is proposed critical habitat listed for the West Indian 

manatee but it does not occur in this county.

Conclusion: No federally listed threatened or endangered species or species proposed for listing 

are likely to be found at these release sites. If they were to enter the sites, their presence would be 

fleeting as the habitat is either not suitable or does not contain constituent elements required by the 

species. Field activities will result in no changes to the habitat used by any listed species or species 

proposed for listing. The sites are not within or near designated critical habitat or habitat proposed 

for designation. Therefore, the action will have no effect on listed species or species proposed for 

listing and would not affect designated critical habitat or habitat proposed for designation.

Overall Conclusions

The field test sites in these permit applications have been in agricultural or forest research, or in 

agricultural production or forest tree plantations for from 5 to 50 years. No federally listed 

threatened or endangered species or species proposed for listing are likely to be found at any of the 

release sites. If they were to enter the site, their presence would be fleeting as the habitat is either 

not suitable or does not contain constituent elements required by the species. Field activities will 

result in no changes to habitat used by any listed species or species proposed for listing. The sites 

are not within or near designated critical habitat or habitat proposed for designation. Therefore, the 

action will have no effect on listed species or species proposed for listing and would not affect 

designated critical habitat or habitat proposed for designation. Because of this no effect 

determination, consultation and/or the concurrence of the USFWS and/or the NMFS are not 

required. 
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APPENDIX V: Supplemental Permit Conditions

For Release of Eucalyptus grandis x Eucalyptus urophylla under permits 08-011-106rm and 

08-014-101rm

(1) The test sites and adjacent land within 100 meters shall be monitored for any volunteer 

Eucalyptus plants every 6 months during the field test (as indicated in the permit) and for one year 

after completion of the field test, during which time any volunteer plants will be destroyed before 

they flower. During the monitoring period following completion of the field test, the site will not be 

planted with Eucalyptus, so that any volunteer seedlings that emerge can be easily identified. If 

volunteers or stump sprouts are still emerging at the end of the first year, a second year will be 

added to the monitoring period to ensure that no shoots are continuing to be produced.

(2) Please note that transportation of all test and plant materials to and from the field test location 

must be done in accordance with APHIS/USDA regulations outlined in "Container requirements 

for the movement of regulated articles", 7CFR 340.8(b) unless a shipping container variance has 

been approved by APHIS-BRS.

(3) BRS should be notified in writing of any proposed changes to the permit application (or 

approved permit) including for example confinement protocols, transgenic lines or constructs, 

release locations, acreage, etc. Changes usually require amendments to the permit and must be preapproved by BRS. Requests should be directed to Regulatory Permit Specialist, USDA APHIS 

BRS, Biotechnology Permit Services, 4700 River Road, Unit 91, Riverdale, Maryland 20737.

(4) Any regulated article introduced not in compliance with the requirements of 7 CFR part 340 or 

any standard or supplemental permit conditions, shall be subject to the immediate application of 

such remedial measures or safeguards as an inspector determines necessary, to prevent the 

introduction of such plant pests. The responsible party may be subject to fines or penalties as 

authorized by the Plant Protection Act (7 U.S.C. 7701-7772).

(5) This Permit does not eliminate the permittee's legal responsibility to obtain all necessary 

Federal and State approvals, including for the use of: (1) any non-genetically engineered plant pests 

or pathogens as challenge inoculum; (2) plants, plant parts or seeds which are under existing 

Federal or State quarantine or restricted use; (3) experimental use of unregistered chemical; and (4) 

food or feed use of genetically engineered crops harvested from the field experiment.

(6) APHIS/BRS and/or an APHIS/PPQ personnel may conduct inspections of the test location, 

facilities, and/or records at any time.

(7) Harvested plant material may not be used for food or animal feed unless it is first devitalized 

and approved for such use by the U.S. Food and Drug Administration; and for plant-incorporated 

protectants, a tolerance for the pesticide must first be established by the U.S. Environmental 

Protection Agency.
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(8) The permittee shall provide all workers with identifying characteristics of the threatened 

Eastern indigo snake and instructions on what to do if the species is encountered. This shall be 

done for release sites within the known range of the snake which include the following counties: 

Baldwin AL, Evans GA, Bay FL, Highlands FL, Marion FL, Gadsden FL, Columbia, FL and 

Taylor FL.

(9) Reporting an Unauthorized or Accidental Release

1. According to the regulation in 7 CFR 340.4(f)(10), APHIS shall be notified orally immediately 

upon discovery and notified in writing within 24 hours in the event of any accidental or 

unauthorized release of the regulated article.

- For immediate verbal notification, contact APHIS BRS Compliance Staff at (301) 734-5690 and 

ask to speak to a Compliance and Inspection staff member. Leave a verbal report on voicemail if 

the phone is not answered by a Compliance Officer.

- In addition, in the event of an emergency in which you need to speak immediately to APHIS 

personnel regarding the situation, you may call:

The APHIS/BRS Regional Biotechnologist assigned in the region where the field test occurs:

For Western Region, contact the Western Region Biotechnologist at (970) 494-7513

or e-mail: BRSWRBT@aphis.usda.gov

For Eastern Region, contact the Eastern Region Biotechnologist at (919) 855-7622 or e-mail:

BRSERBT@aphis.usda.gov

Or

The APHIS State Plant Health Director for the state where the unauthorized release occurred. The 

list of APHIS State Plant Health Directors is available at:

http://www.aphis.usda.gov/services/report_pest_disease/report_pest_disease.shtml.

or http://pest.ceris.purdue.edu/stateselect.html

2. Written notification should be sent by one of the following means:

By e-mail:

BRSCompliance@aphis.usda.gov

By mail:

Biotechnology Regulatory Services (BRS)

Compliance and Inspection Branch

USDA/APHIS

4700 River Rd. Unit 91

Riverdale, MD 20737

3. Additional instructions for reporting compliance incidents may be found at

http://www.aphis.usda.gov/biotechnology/compliance_incident.shtml

(10) Reporting Unintended Effects:

According to the regulation in 7 CFR 340.4(f)(10)(ii), APHIS shall be notified in writing as soon as 

possible but within 5 working days if the regulated article or associated host organism is found to 

have characteristics substantially different from those listed in the permit application or suffers any 
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unusual occurrence (excessive mortality or morbidity, or unanticipated effect on non-target 

organisms).

Written notification should be sent by one of the following means:

By e-mail:

BRSCompliance@aphis.usda.gov

By mail:

Biotechnology Regulatory Services (BRS)

Compliance and Inspection Branch

USDA/APHIS

4700 River Rd. Unit 147

Riverdale, MD 20737

(11) Reports and Notices:

Send notices and all reports (CBI and CBI-deleted or non-CBI copies) to BRS by e-mail, mail, or 

fax.

BRS E-mail:

BRSCompliance@aphis.usda.gov

BRS Mail:

Animal and Plant Health Inspection Service (APHIS)

Biotechnology Regulatory Services (BRS)

Compliance and Inspection Branch

4700 River Rd. Unit 91

Riverdale, MD 20737

BRS Fax:

Compliance and Inspection Branch

(301) 734-8669

a. Planting Report

Within 28 calendar days after planting, submit a report, in paper format or electronically, that 

includes the following information for each field test location:

i. Permit number;

ii. Regulated article;

iii. Release location [provide state, county, internal identification number (if available), and either a 

single GPS coordinate as a reference point (center of plot or specify corner) or specific address];

iv. List of all constructs and specific transformed lines (event) planted;

v. Total acreage of regulated article planted;

vi. Total acreage of any border rows planted;

vii. The actual planting date(s)

If multiple plantings occur that are separated in time by more than a month, then a planting report is 

required within 28 days of each planting.

b. Annual Report

Within 30 days after the anniversary date (one year increments from the effective date) an Annual 

Report must be submitted to APHIS. FAILURE TO SUBMIT ANNUAL REPORTS MAY 

RESULT IN REVOCATION OF THE PERMIT. The Annual Report shall reflect the current status 
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and observations to date for each location. It shall include the information submitted in the Planting 

Report, plus the following:

i. An accounting of the acreage or number of plants per line (event) for each construct that remain 

in the ground;

ii. A detailed map of the plantings;

iii. Total remaining acreage (include acreage of border rows if appropriate);

iv. The methods of observation;

v. The resulting data and analysis regarding all deleterious effects on plants, non-target organisms, 

or the environment. This should include, but not be limited to, data on insect damage, disease 

susceptibility, gross morphology and any indications of weediness;

vi. If any material was harvested, removed, or terminated or otherwise destroyed, a disposition 

table with the following information for each line (event) released should be provided:

date(s) of harvest, removal, and/or termination; a formal record of how the regulated material was 

removed from the environment; what material and how much was harvested or removed and where 

it was transported, stored and further processed up to the time it is or was to be taken to a contained 

facility; and what was done to devitalize residual and/or harvested material at the location.

In this report also provide data documenting which trees produced flowers and which if any 

produced viable seeds. Also document seedling volunteer monitoring, including any volunteers 

found and the method of devitalization.

c. Field Test Report

Within 6 months after the expiration date of the permit, the permittee is required to submit a Field 

Test Report.

NOTE: If a new application is approved to continue the field test past its scheduled expiration date, 

an annual report should continue to be submitted until the final expiration date, at which point the 

Field Test Report will be due after 6 months. Field Test Reports provide the final status and 

observations at each location and shall include:

i. List of all constructs and specific transformed lines (event) planted;

ii. Planting date(s), and harvest dates if any material was harvested;

iii. Total acreage of regulated article planted;

iv. Total acreage of any border rows planted;

vi. The methods of observation;

vii. The resulting data and analysis regarding all deleterious effects on plants, non-target organisms, 

or the environment. This should include, but not be limited to, data on insect damage, disease 

susceptibility, gross morphology and any indications of weediness.

viii. A disposition table with the following information:

Site name (or GPS), crop, harvest date(s), and disposition of harvested material. Date(s) of harvest, 

removal, and/or termination; a formal record of how the regulated material was removed from the 

environment; what material and how much was harvested or removed and where it was transported, 

stored and further processed up to the time it was taken to a contained facility; and what was done 

to devitalize residual and/or harvested material at the location.

We encourage the inclusion of other types of data if the applicant anticipates submission of a 

petition for determination of non-regulated status for their regulated article. APHIS considers these 

data reports as critical to our assessment of plant pest risk and development of regulatory policies 
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based on the best scientific evidence. Failure by an applicant to provide data reports in a timely 

manner for a field trial may result in the withholding of permission by APHIS for future field trials.

d. Flowering monitoring report

In the locations where there are other species of eucalyptus within 1000 meters of the test plots, if 

there is any overlap in flowering between the transgenic trial and the non-transgenic trees, this must 

be reported to APHIS. 

e. Monitoring Report

The final monitoring report is due no later than 2 months from the end of the volunteer monitoring 

period.

The report must include:

i. Dates when the field location and perimeter fallow zone were inspected for volunteer plants;

ii. Number of volunteers observed;

iii. Any actions taken to remove or destroy volunteers.