1 

# Ecological Risk Assessment for the Proposed New Chemical Registration of GS

- U

- ACTX

- Hv1a

- SEQ2 (VersutideTM peptide) on Vegetables and Cole Crops; Herbs, Spices, and Mints; Pasture and Hay Crops; Fruit, Nut, and Vine Crops;

Field Crops; Commercial Flowers and Ornamental Plants; Forest, Shade Tree, and Nursery Stock; and Turf

ASSOCIATED BARCODE : D 4 1 1 5 0 7

By Meghan Radtke, Ph.D., Biologist

Peer Reviewers Faruque Khan, Ph.D., Senior Scientist

Edward Odenkirch e n, Ph.D., Senior Scientist Sujatha Sankula, Ph.D., Lead Biologist

Acting Branch Chief

Edward Odenkirchen , Ph.D.

Date of Approval May 3 0 , 20132 

### 1. PROBLEM FORMULATION

1.1 Nature of the Registration Action

This environmental risk assessment evaluates the potential ecological risks of the new broad

spectrum insecticide, GS

- U

- ACTX

- Hv1a

- SEQ2 (VersutideTM peptide) . The insecticide is proposed for use on a number of agricultural and home/gar den uses; general categories include: vegetable and cole crops; herbs, spices, and mints; fruit, nut, and vine crops; field crops; commercial flowers and ornamental plants; forest, shade tree, and nursery stock; and turf. The

insecticide is listed as bei ng effective to control numerous insects including: alfalfa looper, army worm, Colorado potato beetle larvae, hornworms, spotted cucumber beetle, tobacco budworm, European corn borer, cankerworm, western tent caterpillar, cutworms, corn earworm, and gypsy

moth. T wo formulations ( 20% and 30% ai) are being registered. The proposed maximum single application rate is 0.8 lb ai/A with a maximum yearly application rate (assumed) of 0.8 lb

ai/A. The products are foliar sprays that are applied via ground or a erial application equipment.

1.2 Nature of the Chemical Stressor

GS

- U

- ACTX

- Hv1a

- SEQ2 is a peptide that is derived from the venom of a species of Australian funnel spider ( Hadronyche versuta ). Specifically, it is derived from the u

- ACTX

- HV1 peptide, which represents a small fraction of the funnel spider’s veno m (< 0.5% by dry weight); there are only two amino acid differences between the u

- ACTX

- HV1 peptide and GS

- U

- ACTX

- Hv1a

SEQ2. The protein structure consists of a solvent

- accessible beta

- hairpin pr otruding from a disulphide

- bonded globular core comprising four beta

- turns. The three int ramolecular disulphide

bonds for m a cystine knot motif similar to that seen in several other neurotoxic peptides. The u

ACTX

- HV1 peptide acts by inhibiting insect vo ltage

- gated calcium channel currents (Fletcher et
al. 1997) and by inhibiting the calcium activated potassium channels (Gunning et al., 2008)

. GS

- U

- ACT X

- Hv1a

- SEQ2 is mass produced via transgenic yeast.

1.3 Use Characterization

GS

- U

- ACTX

- Hv1a

- SEQ2 is pro posed for registration on a wide variety of uses.

Vegetable and Cole Crops : artichoke, arugula, asparagus, beans, beets, bok choy, broccoli, broccoli raab, Brussels sprouts, cabbage, cardoni, carrots, cassava, cauliflower, cavalo, celeriac, celery, chayote root, chervil, chick peas, chicory, China greens, Chinese artichoke, Chinese broccoli, Chinese cabbage (napa), Chinese mustard cabbage, collards, corn salad, cress, cucumber, cucurbits, dandelion dock, dry bulb onions, edible burdock, edible chrysa nthemum, edible gourds, eggplant, escarole, endive, garlic, gin g er, ginseng, green onions, greens (beet, China, dandelion, mustard, turnip, rape), ground cherry, ho rseradish, Jerusalem artichoke, kale, kohlrabi, leeks, lentils, lettuce (head, leaf, romaine ), lupine, malanga, melons , mizuna, mustard spinach, okra, olives, onions, parsley, parsnips, peas, pepino, peppers, potatoes, pumpkins, purslane, radicchio, radishes, rhubarb, rutabaga, salsify, shallots, skirret, soybean foliage, soybean, spinach, squash , sugar beets, sweet potatoes, swiss chard, tomatillo, tomatoes, turmeric, turnips, turnip

- rooted chervil, turnip

- rooted parsley, watercress, yams3 

### Herbs, Spices, and Mints: angelica, balm, basil, borage, burnet, catnip, chamomile, chervil, chives, cilantro, clary, coriander, costmary, curry, dill, dill weed , fennel, horehound, hyssop, lavender, lemongrass, lovage, marjoram, mint, nasturtium, oregano, parsley, peppermint, rosemary, sage, savory, spearmint, sweet bay, tansy, tarragon, thyme, wintergree n, woodruff,

wormwood

Pasture and Hay Crops: alfalfa (hay and seed), hay and other forage crops, pasture (grasses and hay), silage

Pome and Stone Fruit Trees : apples, apricots, crabapple, cherries, figs, loquat, mayhaw, nectarines, peaches, pears, plum s, prunes, quince

Nut Trees: almonds, cashew, chestnuts, filberts, macadamia nut, pecans, pistachios, walnuts

Tree Fruits: acerola, atemoya, biriba, black sapote, canistel, cherimoya, custard apple, feijoa, guava, llama, jaboticaba, kiwi, longan, lyche e, mamey sapote, mango, papaya, passion fruit

, pulasan, rambutan, sapodilla, soursop, Spanish lime, star apple, starfruit, sugar apple, ti palm, wax jambu, white sapote

Citrus: grapefruit, lemons, limes, organs, tangerines

Small Fruit and Berries: black berries, blueberries, boysenberries, cranberries, currants, elderberry, gooseberry, huckleberry, juneberry, ligonberry, loganberries, raspberries, salal, strawberries, grapes

Tropical and Other Fruit: bananas, kiwi, persimmon, pomegranate, pineapple

Tro pical/Sub

- Tropical Fruits: avocado, guava, lychee, sugar apple

Field Crops: canola/rape seed, evening primrose, meado w foam, corn (field, sweet, popcorn, seed), cotton, hops, jojoba, peanuts, rice safflower

Small Grains: barley, buckwheat, grain amara nth, millet, milo, oats, pearl, proso millet, rye, triticale, wheat, sorghum, soybeans, sunflower (oil and seed), tobacco, coffee

Commercial Flowers and Ornamental Plants: bedding plants, flowers (greenhouse and field), greenhouse ornamentals, greenhouse vegetables, container stock

Forest, Shade Tree, and Nursery Stock: deciduous, forest, shade trees, nursery trees, ornamental trees, conifers (including Christmas trees)

Turf: turf and turf grown for seed

According to the label

, for all uses, a single application can range from 0.025 to 0.8 lb ai/A; the single maximum application rate is 0.8 lb ai/A. The target pests include: achema sphinx moth, alfalfa caterpillar, alfalfa looper, alfalfa webworm, amorbia, armyworm, artichoke plume moth,4 

### asp aragus beetle, avocado leaf roller, azalea caterpillar, azalea moth, bagworm, banana skipper, banana moth, banded sunflower moth, beet armyworm, bertha armyworm, blackheaded budworm, blackheaded fireworm, blueberry leaf roller, bollworm, browntail moth, ca bbage budworm, cabbage looper, cabbage webworm, California oakworm, cankerworm, celery leaftier, cereal leaf beetle, cherry fruitworm, citrus cutworm, citrus peelminer, codling moth, Colorado potato beetle larvae, corn earworm, corn rootworm, cotton leaf p erforator, cottonwood leaf beetle, cranberry fruitworm, cranberry girdler, cross

- stripped cabbageworm, cutworms, diamondback moth, douglas fir tussock moth, eastern tent caterpillar, ello moth, elm spanworm, European corn bor er, European grapevine moth, E uropean skipper, fall webworm, filbert leaf roller, filbert webworm, fireworm, Florida fern caterpillar, fruit tree leaf roller, grape berry moth, grape leaffolder, grapeleaf skeletonizer, green cloverworm, green fruitworm, greenstriped mapleworm, gummoso s

- batrachedra commosae, gypsy moth, headworm, heliothis, hemlock looper, hickory shuckworm, hornworm, imported cabbageworm, jack pine budworm, leaf rollers, light brown apple moth, lo moth, looper, melonworm, mimosa webworm, navel orangeworm, oblique band ed leaf roller, oleander moth, omniforous leaftier, omnivorous leaf roller, orange tortix, orangedog, oriental fruit moth, pandemic leaf roller, peach twig borer, pecan nut casebearer, pickleworm, pine butterfly, pine tip moths, plum curculio, podworm, red

- neck

peanut worm, redbanded leaf roller, redhumped caterpillar, rindworm complex, roughskinned

cutworm, saddleback caterpillar, saddle prominent caterpillar, saltmarsh caterpillar, silverleaf whitefly nymphs, sod webworm, southwestern corn border, soybean looper, spanworm, sparganothis, spotted asparagus beetle, spotted cucumber beetle, spotted cutworm, spring and fall cankerworm, spruce budworm, striped cucumber beetle, sunflower moth, tent caterpillar, thecla

thecla basilides, tobacco budworm, tobacco hornworm, tomato fruitworm, tomato hornworm, tomato pinworm, tortix, tortix moth, tropical sod webworm, tufted apple budmoth, tussock moth, twig borer, variegated cutworm, variegated leaf roller, velvetbean caterpillar

, viburnam beetle,

walnut cate rpillar, webworm, western bean cutworm, western corn cutworm, western raspberry fruitworm, western tent caterpillar, western tussock moth, yellow margined leaf beetle larvae,

and yellow s triped armyworm.

The label does not specify a yearly maximum applicat ion rate. Therefore, it is assumed that 0.8

lb ai/A is the maximum annual application rate for GS

- U

- ACTX

- Hv1a

- SEQ2 . For single application rates below 0.8 lb ai/A, the application interval is 3 to 10 days.

1.5 Analysis Plan

The screening

- level risk ass essment is used as the approach to determine risk to non

- target organisms from the proposed new uses of GS

- U

- ACTX

- Hv1a

- SEQ2

. Given that the toxicity

data for this chemical are non

- definitive and that limited fate data are provided, the assessment will qualitatively analyze the anticipated effects of GS

- U

- ACTX

- Hv1a

- SEQ2 on non

- target organisms in the environment. The qualitative analysis is done by comparing the estimated environmental concentrations (EECs) w ith the toxicity value generated during testing. The environmental fate of proteins will also be explored.

2. ENVIRONMENTAL FATE CHARACTERIZATION

Limited f a te data were submitted in support of the registration of GS

- U

- ACTX

- Hv1a

- SEQ2.

The active ingredie nt is a peptide derived from one of the toxicologically active components of5 

### Australia n funnel spider venom (Figure 1)

. Its molecular weight is 4565 kDa and the pH is 4.89

- 5.95 (MRIDs 48779002 and 48779101)

. It is a large molecule that is made up of a chain of 41

amino acids. Its surface is covered in hydrophilic residues

, making it h ighly soluble in water , but

insoluble in ethanol (specific solubility data were not submitted) . The formulation contain s

pa racellular transport mechanisms (derived from inert ingredients on EPA’s list) to facilitate the translocation of the peptide from the gut into the insect’s hemocoel and ultimately the nervous

system

. Upon entering the environment, GS

- U

- ACTX

- Hv1a

- SEQ2 is readily biodegradable and

breaks down into its amino acid components. Its conformation (quaternary structure) determines its toxicity; thus, even a minor change to its structure may render it non

- toxic. Its terrestrial half

life is approximately 4 days

, extrapolated from simulated sunlight exposures in terrestrial settings where degradation occurred after 48

- hours (MRID s 48779002 and 48779003).

Figure 1. Richardson schematic of calculated 3

- D structures of u

- ACTX

- Hv1a peptide

from Protein Data Bank number 2H1Z (MRID 48779001)

The following required fate guideline studies, as stipulated by the Code of Federal Regulation, are not available and are consequently identified as data gaps:

• 835.2129 (hydrolysis)

• 835.2240 (photodegradation in water)

• 835.241 0 (photodegradation in soil)

• 835.2370 (photodegradation in air

– conditional, depending on chemical’s properties)

• 835.4100 (aerobic soil metabolism)

• 835.4200 (anaerobic soil metabolism)

• 835.4300 (aerobic aquatic metabolism)

• 835.4400 (anaerobic aquatic metabolism)

• 835.1230/835.1240 (leaching ad sorption/desorption)

• 835.6100 (terrestrial field dissipation)

• 835.1410 (volatility laboratory

– conditional, depending on chemical’s properties)6 

### 3. EXPOSURE ESTIMATION

3.2 Aquatic Exposure Modeling

Given the lim ited amount of data available for this assessment, a simplified approach will be taken to estimate environmental exposure. The standard pond used in the Tier I GENEEC or

Tier II the Pesticide Root Zone Model, (PRZM, Carousel et al . 1997) and the Exposure Analysis Modeling System (EXAMS, Burns 2000) is 1 ha in size and cont ains 20,000,000 L of water. Rather than using the models to calculate a n estimated environmental concentration ( EEC ) , the

EEC is calculated by dividing the amount of GS

- U

- ACTX

- Hv1a

- SEQ2 that is applied within a 1 ha area by the 20,000,000 L contained in the pond. The application rate is 0.8 lb ai/A. This is converted to mg/ha by:

(0.8 lb/A )(362874 mg/lb)(1 A/0.404686 ha) = 896

, 6 80 mg/ha

Dividing 896,680 mg by 20,000,000 L yields 0.044 mg/ L , which is the concentration of GS

- U

ACTX

- Hv1a

- SEQ2 that is expected in the standard pond. This represents the peak concentration in the pond; concentrations over longer periods of time are ex pected to be much lower.

This approach is considered conservative because the pesticide is being directly applied to the

water rat her than arriving in the pond via runoff . Furthermore, the pond does not have any inputs or outputs that would allow for exc hange of water.

3.3 Terrestrial Exposure

Terrestrial wildlife exposure estimates are typically calculated for birds and mammals emphasizing a dietary exposure route for uptake of pesticide residues on vegetative matter and

insects. Terrestrial wildlife exposure estimates are c alculated with T

- REX v1.5 .1 model (August 20, 2012) . The residues, or e stimated environmental c oncentrations (EECs), on food items are compared with sub

- acute dietary toxicity data , acute oral data, and chronic dietary or oral toxicity data. Foliar application exposures are estimated for birds in Table 1 and mammals in

Table 2

. In this case, a single application rate of 0.8 lb ai/A was modeled. The risk assessment uses a default foliar dissipation half

- life estimate of 35 day s. This default value is used in lieu of representative foliar dissipation data for GS

- U

- ACTX

- Hv1a

- SEQ2 , because no suitable data were provided that evaluated foliar dissipation (although a 4

- day terrestrial half

- life is provided,

it is unclear if it is a foliar dissipation half

- life or if it is for another medium)

.7 

### Table 1 . Avian EECs for the Proposed GS

- U

- ACTX

- Hv1a

- SEQ2 Uses

Feeding Category Dietary

- based
EECs (mg/kg

food item) Dose

- based EECs (mg/kg

- bw)

Small
(20 g) Medium
(100 g) Large
(1000 g)

All Uses
1 app at 0. 8 lb ai/A

Short grass 192.00 218.67 124.69 55.83 Tall grass 88.00 100.22 57.15 25.59 Broadleaf plants 108.00 123.00 70.14 31.40 Fruits/pods 12.00 13.67 7.79 3.49 Arthropods 75.20 85.65 48.84 21.87

Seeds

- 3.04 1.73 0.78



### Table 2 . Mammal EECs for the Proposed GS

- U

- ACTX

- Hv1a

- SEQ2 Uses

Feeding Category Dietary

- based
EECs (mg/kg

food item) Dose

- based EECs (mg/kg

- bw)

Small
(15 g) Medium
(135 g) Large
(1000 g)

All Uses
1 app at 0.8 lb ai/A

Short grass 192.00 183.06 126.52 29.33 Tall grass 88.00 83.90 57.99 13.44 Broadleaf plants 108.00 102.97 71.17 16.50 Fruits/pods 12.00 11.44 7.91 1.83 Arthropods 75.20 71.70 49.55 11.49

Seeds

- 2.54 1.76 0.41



### For insects, only two EECs are calculated. The tall grass EEC represents the exposure through dietary intake whereas the arthropod EEC represents exposure through contact (Tabl e 3 ) ( see USEPA 2012 for further details about the EECs chosen to represent honeybee exposures ). Given that honeybee larval toxicity data were not available , only GS

- U

- ACTX

- Hv1a

- SEQ2 doses for

adult bees are considered.

Table 3 . Adult Insect EEC s for the Proposed GS

- U

- ACTX

- Hv1a

- SEQ2 Uses

Use EEC based on tall grass mg/kg

- diet (dose in μg ai/bee*) EEC based on arthropod mg/kg

- diet (dose in μg ai/bee**) 1

All uses ( 1 app at 0. 8 ai/A ) 88.00 (25.7 ) 75.20 (9.6 )
*Conversion made by multiplying EEC by food consumption rate of adult bee (0.292 g/day) **Conversion made by multiplying EEC by body weight of adult bee (0.128 g)



### Terrestrial and semi

- aquatic plants may be ex posed to pesticides from runoff and spray drift.

Semi

- aquatic plants are those that inhabit low

- lying wet areas that may be dry at certain times of the year. Given that the solubility of GS

- U

- ACTX

- Hv1a

- SEQ2 is not available, TerrPlant (v.1.2.1 ) cannot be used to calcul ate EECs for terrestrial plants

. In lieu of this, the risks to plants8 

### will be qualitatively assessed using toxicity information from the vegetative vigor and seedling emergence limit tests.

4. ECOLOGICAL EFFECTS CHARACTERIZATION

E cotoxicity data are limited to acute mammal, acute avian oral , acute freshwater fish, acute freshwater invertebrate, acute honeybee contact, terrestrial plant seedling emergence, and terrestrial plant vegetative vigo r for GS

- U

- ACTX

- Hv1a

- SEQ2. According t o the Federal Code of Regulations, a number of studies are missing and are considered data gaps

. Below is a list of the studies and a description of the assumptions that were made in this risk assessment in the absence of data. Filling the data gaps woul d reduce the uncertainty associated with the assumptions that were made.

• 850.1075 (acute estuarine/marine fish toxicity)

– acute freshwater fish toxicity data were

used in lieu of estuarine/marine fish data.

• 850.1025/850.1035 (acute estuarine/marine inver tebrate toxicity)

– acute freshwater

invertebrate data were used in lieu of estuarine/marine invertebrate data.

• 850.1300 (freshwater invertebrate life cycle test)

– exposure was presumed to be negli g i ble because the peptide is expected to immediately break d own in aquatic environments, thus, long

- term exposures are unlikely.

• 850.1350 (estuarine/marine invertebrate life cycle test)

– exposure was presumed to be negligible because the peptide is expected to immediately break down in aquatic environments, thus, long

- term exposures are unlikely.

• 850.1400 (freshwater fish early life stage test)

– exposure was presumed to be negligible because the peptide is expected to immediately break down in aquatic environments, thus, long

- term exposures are unlikely.

• 850.1400 (estuarine/marine fish early life stage test)

– exposure was presumed to be negligible because the peptide is expected to immediately break down in aquatic environments, thus, long

- term exposures are unlikely.

• 850.2100 (acute avian oral toxicity with malla rd and passerine species)

– acute oral data from the bobwhite quail was used as a surrogate for the mallard and passerine species.

• 850.2200 (acute avian dietary toxicity)

– acute oral data from the bobwhite quail was used as a surrogate.

• 850.2300 (avian reproduction toxicity)

– information regarding the lack of toxicity to vertebrates was used to support the assessment.

• 870.3800 (mammalian reproduction toxicity)

– information regarding the lack of toxicity to vertebrates was used to support the assessment

.

• 850.4100 (seedling emergence)

– reliable information was not available for soybean. Data from the other tested dicots were used as surrogates.

• 850.4150 (vegetative vigor)

– reliable information was not available for soybean. Data

from the other tested dicots were used as surrogates.

• 850.4400 (aquatic plant growth

– vascular plants)

– lack of effects in terrestrial plants and presumed negligible exposure because the peptide is expected to immediately break down in aquatic environments were used to suppor t the assessment.9 

### • 850.4500 (aquatic plant growth

– non

- vascular plants)

– lack of effects in terrestrial plants and presumed negligible exposure because the peptide is expected to immediately break down in aquatic environments were used to support the asse ssment.

In addition, a honeybee oral toxicity study (non

- guideline, OECD 213 ) and larval toxicity (non

guideline) woul d also be requested to evaluate toxicity to honeybees and non

- listed terrestrial

invertebrates. In the absence of these data, risks are assumed to honeybees because the pesticide is designated as an insecticide.

Aquatic Organisms

Acute data for freshwater fish and invertebrates are available (Table 4 ) . For the freshwater fish study, a limit test (100 mg ai/L) was performed with rainbow trout . The treatment group exhibited 10% mortality; however, this is considered normal background mortality ( USEPA

1996a ) . No sub

- lethal effects were observed and the LC 50 

### was determined to be above the 100 mg ai/L concentration.

The acute freshwater invertebrate limit test was performed with the waterflea. Waterfleas were exposed to a single concentration of 1000 mg ai/L. Mortality (3 to 7 %) occurred in both the control and treatment groups, but it was at rates consistent with nor mal background mortality (USEPA 1996b) . No sub

- lethal effects were observed and the EC 50 

### was determined to be above the 1000 mg ai/L concentration.

One uncertainty for both the fish and aquatic invertebrate studies is the stability of GS

- U

- ACTX

Hv1a

- SEQ2 in water . Understanding the stability of a compound is important because it directly affects the exposure concentration in the test. The half

- life of the peptide in terrestrial habitats is approximately four days. No hydrolysis data have been submitted, thus the concentration of GS

U

- ACTX

- Hv1a

- SEQ2 over the duration of the aquatic tests is unknown ; it is possible that the degradation occurred very rapidly

. I t is assumed that the acute exposures represent real

- life exposure scenarios, regardless of the degradation rate

. It is unimportant whether the peptide degraded quickly upon entering the aquatic environment or persisted through the experiment.

No effects occurred with in the test systems; however, the fish and invertebrate studies are classified as “supplemental” because of the chemical stability uncertainty.

Acute and chronic data are not available for estuarine/marine fish or invertebrates. Likewise,

chronic data are not available for freshwater and estuarine/marine fish or aqu atic inverteb rates.

D ata for aquatic vascular and non

- vascular plants also are not available.10 

### Table 4 . Acute Toxicity Effects of GS

- U

- ACTX

- Hv1a

- SEQ2 o n Aquatic Organisms

Species Study

Guideline A.I. Toxicity Endpoint Study and Toxicity

Classification MRID #

Rainbow Trout (Oncorhynchus mykiss) 850.1075 31.5 % LC 50 > 100 mg ai/A NOAEC = 100 mg ai/L

No sub

- lethal effects Supplemental

Practically

non

- toxic 48779014

Waterflea ( Daphnia

magna ) 850.1010 31.5 % EC 50 > 1000 mg ai/L NOAEC = 1000 mg ai/L

No sub

- lethal effects Supplemental

Practically

non

- toxic 48779015



### Terrestrial Organisms Acute oral toxicity data are available for birds and mammals (Table 5 ) . A limit test was conducted by exposing the bobwhite quail via oral gavage to concentrations of 630 mg ai/kg

- bw.

Ten control birds were tested and ten birds were expos ed to GS

- U

- ACTX

- Hv1a

- SEQ2. No

mortalities or sub

- lethal effects were observed, thus, the LD 50 

### is > 630 mg ai/kg

- bw. Acute dietary data are not available for bird s. Chronic/reproduction data also are not available.

Similarly , an acute oral toxicity study (up/down procedure) was conducted on three rats. Initially, one female rat was administered an oral dose of 5000 mg ai/kg

- bw. When mortality did

not occur and no sub

- lethal effects (including changes to behavior and decreases in body weight) were observed, an additional two rats were dosed with the same concentration. The experiment was terminated when none of the three rats showed any signs of toxicity over a two week period

. The resulting LD 50 

### is > 5000 mg ai/kg

- bw. A chronic two

- generation reproduction study with

rats is not available.

An acute honeybee contact toxicity test was available f or GS

- U

- ACTX

- Hv1a

- SEQ2 (Table 5 ). Sixty bees were exposed to a con centration of 25 μg ai/bee and 60 bees were exposed to water (controls) via a droplet of solution to the abdomen (contact exposure) . Mortality rates for each group were at or below 8% and no sub

- lethal effects were observed. Consequently, the contact

LD 5 0 

### for honeybees is > 25 μg ai/bee. Acute oral and larval data are not available for honeybees.

Table 5

. Acute Toxicity Effects of GS

- U

- ACTX

- Hv1a

- SEQ2 on Terrestrial Animals

Species Study

Guideline A.I. Toxicity Endpoint Study and Toxicity

Classification MRID #

Bobwhite quail ( Colinus virginianus ) 850.2100 31.5% LD 50 > 630 mg ai/kg

- bw NOAEC = 630 mg ai/kg

- bw

No effects Supplemental

Slightly toxic 48779013

Rat ( Rattus norvegicus ) 870.1100 31.5% LD 50 > 5000 mg ai/kg

- bw NOAEC = 5000 mg ai/kg

- bw Acceptable

Practically

non

- toxic 48779007

Honeybee ( Apis mellifera ) 850.3020 31.5% Contact LD 50 > 25 μ g ai/bee

NOAEC = 25 μ g ai/bee

No sub

- lethal effects Supplemental

Practically

non

- toxic 4877901811 

### Terrestrial plant data were available f or GS

- U

- ACTX

- Hv1a

- SEQ2 (Table 6 ). Seedling emergence and vegetative vigor tests were conducted at application rates of 0.8 lb ai/A. In both studies, the emergence rate for soybean was below 70%, indicating a potential problem with the seed stock. Consequen tly, data for this species are not considered reliable and are not included

in the risk assessment . In the seedling emergence test, none of the species in the treatment group showed reductions in emergence, diameter, len g th, or dry weight that were greate r than 25%. Therefore, the EC 25 

### is > 0.8 lb ai/A for seedling emergence. In the vegetative vigor study, onion and lettuce showed significant decreases (gre at er than 25% in root weight); however these

differences did not translate into above

- ground biomas s. Root weight is generally considered an unreliable endpoint because of the difficulty in obtaining accurate measurements. Effects of 25% or more were not seen for plant height, diameter or dry length; thus, the EC 25 

### is > 0.8 lb ai/A for vegetative vigo r.

Table 6

. Toxicity Effects of GS

- U

- ACTX

- Hv1a

- SEQ2 o n Terrestrial Plants

Species Study

Guideline A.I. Toxicity Endpoint Study

Classification MRID #

Monocots Oat ( Avena sativa ) Ryegrass ( Lolium perenne ) Onion ( Allium cepa ) Corn ( Zea mays )

Dicots Tomato ( Lycopersicon

esculentum ) Cucumber ( Cucumis sativus ) Lettuce ( Lactuca sativa ) Soybean ( Gl y cine max ) Cabbage ( Brassica oleracea ) Carrot ( Daucus carota ) Seedling

emergence

850.4100 31.5 % EC 25 > 0.8 lb ai/A

NOAEC = 0.8 lb ai/A

No effects Supplemental 48779016

Monocots Oat ( Avena sativa ) Ryegrass ( Lolium perenne ) Onion ( Allium cepa ) Corn ( Zea mays )

Dicots Tomato ( Lycopersicon

esculentum ) Cucumber ( Cucumis sativus ) Lettuce ( Lactuca sativa ) Soybean ( Gl y cine max ) Cabbage ( Brassica oleracea ) Carrot ( Daucus carota ) Vegetative vigor

850.4150 31.5 % EC 25 > 0.8 lb ai/A

NOAEC = 0.8 lb ai/A

>25% root weight

decrease in onion and

lettuce that was not

c onsidered reliable Supplemental 48779017



### Incident Database Review

S earch es of the Ecological Incident Information System (EIIS v. 2.1.1 ), Avian Incident Monitoring System (AIMS), and the Incident Database System (IDS) were performed on Ma y 2

,12 

### 2013. No ecological incidents were reported in any of the databases. This is expected given that

GS

- U

- ACTX

- Hv1a

- SEQ2 is a new chemical.

5. RISK DESCRIPTION

Results of the exposure modeling and toxicity studies are used to evaluate the likelihood of adverse ecological effects on non

- target species. For GS

- U

- ACTX

- Hv1a

- SEQ2, this will be a qualitative assessment because of the limited fate and toxicity data.

5.1.1 Risk to Aquatic Organisms

A peak concentration of GS

- U

- ACTX

- Hv1a

- SEQ2 was generated by calculating the amount of active ingredient distributed per liter within the standard PRZM/EXAMS pond. This EEC (0.044 mg ai/L) was compared directly to the toxicity values generated in the regi strant

submitted studies.

Acute Risks to Fish and Invertebrates

The LC 50 

### for freshwater fish is > 100 mg ai/L. Compared to the peak EEC of 0.044 mg ai/L, the

LC 50 

### is at least four orders of magnitude higher. This indicates that risk concerns to freshwater fish on an acute exposure basis are unlikely. Acute toxicity data were not available for

estuarine/marine fish . In lieu of this, freshwater fish are used as a surrogate. This adds some uncertainty to the analysis; however, given that no effec ts (including sub

- lethal) were seen in freshwater fish and the extremely large difference between the LC 50 

### value and the peak EEC, it is unlikely that estuarine/marine fish would be markedly different in their sensitivity to GS

- U

ACTX

- Hv1a

- SEQ2. Consequen tly, estuarine/marine fish are not expected to be at risk on an acute exposure basis.

The EC 50 

### for freshwater invertebrates is > 1000 mg ai/L. Compared to the peak EEC of 0.044 mg ai/L, the EC 50 

### is at least five orders of magnitude larger. Therefore, acu te risk concerns to freshwater invertebrates are unlikely. Acute toxicity data were not available for estuarine/marine invertebrates, thus, the freshwater invertebrate data were used as a surrogate. For the same reasons identified for fish, the freshwate r invertebrate data are considered a reasonably protective surrogate for estuarine/marine invertebrates . Based on this, acute risk concerns are not expected

for estuarine/marine invertebrates.

Chronic Risks to Fish and Invertebrates Toxicity data were no t available for chronic exposures to fish or invertebrates. One of the uncertainties in the acute aquatic studies was the concern that GS

- U

- ACTX

- Hv1a

- SEQ2 degraded very quickly and that the actual exposure experienced by the test organism was far

lower th an the nominal concentration of active ingredient. Hydrolysis data for the peptide are not available, but it is likely that degradation rapidly occurs in water. The quaternary structure (p rotein folding ) of GS

- U

- ACTX

- Hv1a

- SEQ2 is very complex; conditions that even slightly alter the protein’s configuration (e.g., exposure to water) will interfere with its receptor binding

. Terrestrial data indicate that the half

- life of GS

- U

- ACTX

- Hv1a

- SEQ2 is only 4 days. If a

similar or smaller half

- life occurs in wate r, aquatic organisms would not reasonably be expected to experience chronic exposure conditions. Therefore, risk concerns from chronic exposures are considered unlikely.13 

### Aquatic Plants Toxicity data were not available for aquatic vascular and non

- vascul ar plants. At an application rate of 0.8 lb ai/A, terrestrial plants showed no effects. As with the aquatic invertebrate and fish analysis, if the 0.8 lb ai/A were sprayed directly onto a 1 ha 20,000,000 L pond, the concentration is expected to be 0.044 mg ai/L. E xposures to fish (100 mg ai/L) and aquatic

invertebrates (1000 mg ai/L) at much higher rates that 0.044 mg ai/L also showed no effects. Given the specificity of GS

- U

- ACTX

- Hv1a

- SEQ2 to insects , the low probability that aquatic plants have this same receptor, and the peptide’s predicted instability in water, risk concerns for aquatic plants are not expected. To confirm this conclusion, toxicity data for aquatic vascular

and non

- vascular plants could be requested (8 50.4400 and 850.4500).

5.1.2 Risk to Terrestrial Organisms
For terrestrial animals, EECs were generated using T

- REX. These were then compared to the

non

- definitive toxicity values to characterize risks. EECs for GS

- U

- ACTX

- Hv1a

- SEQ2 exposure to terrestri al plants could not be generated because sol ubility data for the protein were not available. Consequently, the discussion of risks to terrestrial plants is based strictly on the toxicity information from the seedling emergence and vegetative vigor studies

.

In addition to the Agency guideline toxicity studies, the registrant submitted several studies with detailed information about the specificity of the GS

- U

- ACTX

- Hv1a

- SEQ2 to insect receptors versus bird/mammal receptors. This information is useful to fu rther characterize the absence of effects in the acute avian and mammal studies as well as to define potential chronic effects.

The GS

- U

- ACTX

- Hv1a

- SEQ2 is closely related to the omega peptide of the same form (50% homology in structure) and exhibits a sim ilar folding pattern (quaternary structure); it also shares homology with the kappa peptide (κ

- ACTX

- Hv1c). It has been documented to share activity on a molecular level with these two other peptides and toxicity information from the omega and kappa peptid es are considered appropriate surrogates (MRID 48779019). In vitro studies indicate that the omega peptide blocks insect calcium channels and affects the ganglionic neurotransmission, but does not inhibit mammalian neuronal calcium channels (Fletcher et a l., 1997). A homolog of the omega peptide with similar insect calcium channel activity was also tested in vertebrates using rat and chick smooth and skeletal nerve

- muscle, respectively. No effects were observed (Chong et al., 2007 ) . The kappa group of p eptides, which shares the calcium activated potassium channel inhibition mode of action with GS

- U

- ACTX

- Hv1a

- SEQ2, also is insect specific and is believed to exhibit little to no activity at rat dorsal root ganglion

neuronal calcium

- activated potassium chan nels (Gunning et al ., 2008). Sollod (2006) demonstrated insect specificity with hybrid peptide compounds

– similar to GS

- U

- ACTX

- Hv1a

SEQ2.

Acute Risks to Birds and Mammals The highest acute dose

- bas ed EEC generated for birds is 2 1 9 mg ai/kg

- bw ( 20 g bird eating short

grass ) . The LD 50 

### of > 630 mg ai/kg

- bw, when compared with the E EC, is three times higher. Thus, acute risks to birds are not expected. Similarly, the acute dose

- based EEC for mammals is 183 mg ai/kg

- bw (15 g mammal eating short grass )

. The corresponding LD 50 

### is > 5000 mg14 

### ai/kg

- bw, which is one order of magnitude higher than the EEC. Therefore, acute risk concerns for mammals are not expected.

Chronic Risks to Birds and Mammals Toxicity data were not available for chronic exposures to birds and mammals. The bioavailability of GS

- U

- ACTX

- Hv1a

- SEQ2 i s expected to be low, based on the physical structure of the peptide. Spider venoms are designed to be injected into their prey. E xp osure through oral means is likely to drastically decrease the toxicity of the compound or eliminate it altogether; peptides are rapidly digested in the gut. As an example, p eptide drugs that have been developed for human oral administration are genera lly limited to a bioavailability of 1

- 2 % because of their molecular size (large), hydrophillicity, and physical stability in the digestive tract (Hamman and Steenekamp, 2011). Consequently, absorption enhancing agents and chemical and physical modificati ons to the peptide molecules are made to increase the absorption. Drugs derived from the cone shell ( Conus fulmen) are based on a peptide with the similar cystine knot of 3 disulfide bonds that is found in GS

- U

- ACTX

- Hv1a

- SEQ2 ; they also share similar physical chemical characteristics. The drug is delivered intrathecally because of a lack of bioavailability through the oral route (Lee, 2010). The limited bioavailability and similarity of the cone shell peptide and GS

- U

- ACTX

- H v1a

- SEQ2 further supports that premise that the oral exposure route poses little chance of toxicity to vertebrates. Consequently, chronic risk concerns are not expected for birds and mammals

.

Acute Risks to Honeybees EECs were generated for both the ora l and contact exposure pathways for honeybees. Toxicological data were available for the contact route; the LD 50 

### is > 25 μg ai/bee for adult honeybees. The contact EEC (converted to a dose) is 9.6 μg ai/bee, which is lower than the

LD 50 

### . Consequently, d irect risks to honeybees via the contact exposure route are not considered likely. Data were not available for the dietary exposure route to adult honeybees nor were toxicity data provided for larvae. Given that GS

- U

- ACTX

- Hv1a

- SEQ2 is an insecticide and is highly specific to insects, risk cannot be precluded. The insecticide is intended to be delivered to its targets through the oral route; t herefore, risk concerns (via ingestion) are identified for honeybees (adult and larval) in the absence of toxicity data. To reduce the uncertainty associated with this assumption, an acute oral toxicity study on adult honeybees (OECD 213) and a larval toxicity study (non

- guideline study) could be performed. In addition, conducting a honeybee toxicity of residues on foliage study (850.3030) would confirm the conclusion of low risk from

hon eybee exposure to aged residues.

Terrestrial Plants Toxicity data for terrestrial plants were available; however the solubility information for GS

- U

ACTX

- Hv1a

- SEQ2 needed to run the TerrPlant model was not . The plant toxicity studies did not exhibit reductions in seedling emergence and vegetative vigor endpoints that were greater than 25%. Given that they were conducted at the highest application rate, effects to plants are prob ably unlikely for the proposed uses. Consequently, no risk concerns are identified for plants from the proposed application rate and uses.15 

### 6

. THREATENED AND ENDANGERED SPECIES CONCERN

Terrestrial insects were the only group of organisms identified to be potentially at risk from the proposed uses of GS

- U

- ACTX

- Hv1a

- SEQ2. To determine whether GS

- U

- ACTX

- Hv1a

- SEQ2 use sites are geographically associated with known locations of listed specie s, a screening

- level

search of the LOCATES (version 2.2.5 ) database was conducted (query performed on

05 / 07 /2013 ). The database compares county

- level location data for listed species with county

level crop production data (as available in the 2007 agricul tural census) to identify any coarse overlaps of listed species with the proposed labeled uses of GS

- UACTX

- Hv1a

- SEQ2

. See Appendix A for a complete list of the crops used in the analysis.

Based on the endangered species analysis

, risk concerns for dire ct effects to terrestrial

invertebrates have been identified (Table 7 ). There is a concern for indirect effects to terrestrial plants that have obligate pollinator requirements or other dependenc ies on terrestrial

invertebrates

. Species that depend on te rrestrial invertebrates for food (amphibians, birds, fish, mammals, reptiles, terrestrial invertebrates) may also be at indirect risk. Appendix A lists th e federally listed species ( 123 species) on which direct effects could potentially occur because of

t he co

- occurrence with crops included in the proposed new uses of GS

- U

- ACTX

- Hv1a

- SEQ2

.

Table 7 . Listed Species Risks Associated with the GS

- U

- ACTX

- Hv1a

- SEQ2 Proposed New

Uses

Listed Taxa Direct Effects Indirect Effects

Terrestrial and semi

- aquatic plants ( monocots and dicots) No Yes

Birds No Yes

Terrestrial phase amphibians No Yes

Reptiles No Yes

Mammals No Yes

Terrestrial in vertebrates Yes (oral) Yes

Aquatic vascular plants No No

Aquatic non

- vascular plants No No

Freshwater fish No Yes

Aquatic phase amphibians No Yes

Freshwater invertebrates No No

Marine/estuarine fish No Yes

Marine/estuarine invertebrates No No



### 7. LITERATURE CITED

Burns, L.A. 2000. Exposure Analysis Modeling System (EXAMS II) User Manual and System

Documentation. EPA/600/R

- 00/081. http://www.epa.gov/ceampubl/16 

### Carousel, R.F., J.C. Imhoff, P.R., Hummel, Cheplick, J.M., A.S. Donigian, and L.A. Suárez.

1997. PRZM

- 3, A Model for Predicting Pesticide and Nitrogen Fate in the Crop Root a nd

Unsaturated Soil Zones: Users Manual for Release 3.12.2. http://www.epa.gov/ceampubl/

Chong Y, Hayes JL, Sollod B, Wen S, Wilson DT, Hains PG, Hodgson WC, Broady KW, King GF, Nicholson GM. 2007. The ω

- atracotoxins: Selective blockers of insect M

- LVA

a nd HVA calcium channels. Biochemical Pharmacology 74:623

- 638.

Fletcher JI, Smith R, O’Donoghue SI, Nilges M. Connor M, Howden MEH, Christie MJ, King GF. 1997. The structure of a novel insecticidal neurotoxin, ω

- atracotoxin

- HV1, from the venom

of an Austra lian funnel web spider. Nature Structural Biology

, 4(7)559

- 566.

Gunning, S. J., F. Maggio, M. J. Windley, S. M. Valenzuela, G. F. King, and G. M. Nicholson.

2008. The Janus

- faced atracotoxins are specific blockers of invertebrate K Ca 

### channels. The FEBS Journal, 275: 4045

- 4059.

Hamman JH, and Steenekamp JH. 2011. Oral peptide drug delivery: Strategies to overcome challenges. In: Peptide Drug Discovery and Development Ed, MC Castanho and NC Santos. Wiley

- VCH, Verlag GmbH& Co KGaA.

Lee S, Kim Y, B S K, Choi HW, Lee JY, Jung HH, Ryu JH, Suh HW, Na HS, Kim HJ, Rhim H, Kim JI. 2010. Analgesic effect of highly reversible ω

- conotoxin FIVA on N type Ca 2+ 

### channels. Molecular Pain . 6;97.

Sollod, B.L. 2006. From venoms to insecticides: exploring the struct ure, function, and evolution of peptide toxins found in the venom of Australian funnel

- web spiders. Ph.D. Thesis, University of Connecticut.

USEPA. 1996a. Ecological effects test guidelines: OPPTS 850.1075 fish acute toxicity test, freshwater and marine. Office of Chemical Safety and Pollution Prevention, United States Environmental Protection Agency.

USEPA. 1996b. Ecological effects test guidelines: OPPTS 850.1 010 aquatic invertebrate acute toxicity test, freshwater daphnids. Office of Chemical Safety and Pollution Prevention, United States Environmental Protection Agency.

USEPA. 2012 . White paper in support of the proposed risk assessment process for bees. Environmental Protection Agency, Health Canada, and California Department of Pesticide Regulation17 

# APPENDIX A : LOCATES Endangered Species Reporting

Crops modeled include: alfalfa hay, almonds, apples, apricots, artichokes

- excluding Jerusalem, asparagus, avocados, bananas, barley for gain, beans

- all other, beans

- dry edible excluding limas, beans

- dry lima, beans

- green lima, beans

- snap (green), beans

- yard long, beets, blackberries and dewberries, blueberries

- tame, b lueberries

- wild, boysenberries, broccoli, Brussels sprouts, buckwheat, cabbage

- chinese, cabbage

- head, cabbage

- mustard, canola, cantaloupes, carrots, cassava, cauliflower, celery, cherries

- sweet, cherries

- tart, chestnuts, chicory, Christmas trees,

Christmas trees cut, citrus fruit

- all, coffee, collards, corn for grain, corn for silage of greenchop, corn

- popcorn and

sweet corn, corn

- sweet for seed, cotton

- all, cotton

- pima, cotton

- upland, cranberries, cropland used only for pasture or grazing, cucumbers and pi ckles, currents, dill for oil, dry beans, eggplant, escarole and endive, field and grass seed crops

- all, figs, floriculture crops, forage

- land used for all hay/haylage grass silage and greenchop, garlic, ginger root, ginseng, grapefruit, grapes, guavas , ha y

- all hay including alfalfa oth er tame small grain and wild, haylage (grass silage and greenchop), haylage or greenchop from alfalfa or alfalfa mixtures, hay

- other tame dry, hay

- wild dry, hazelnuts (filberts), herb and spice plants harvested, herbs

- dried, herbs

- freshcut, honeydew melons, hops, horseradish, jojoba harvested, kale, kiwifruit, lemons, lemons and limes, lentils, lettuce

- all, lettuce

- head, lettuce

leaf, lettuce

- romaine, limes, loganberries, macadamia nuts, mangoes, mangos, mint for oil

- all, mint for oil

peppermint, mint for oil

- spearmint, muskmelons and pepinos, mustard greens, nectarines, oats for grain, okra, olives, onions

- dry, onions

- green, oranges

- all, oranges

- valencia, orchards, other berries, other haylage grass silage and greenchop, other oranges, other tame hay, papayas, parsley, passion fruit, peaches

- all, peaches

- clingstone, peaches

- freestone, peanuts for nuts, pears

- all, pears

- bartlett, pears

- other, peas

- austrian winter, peas

- chinese (sugar and snow), peas

- dry edible, peas

- dry southern (cowpeas), peas

- pigeon, peas

- green (excluding southern), peas

- green southern (cowpeas)

– blackeyed, crowder, etc., pecans

- all, pecans

- improved, pecans

- native and seedling, peppers harvested, peppers other than bell (including chile), peppers

- bell (exclud ing pimientos), permanent pasture and rangeland, other than cropland and woodland pastured, persimmons, pineapples harvested, pineapples not harvested, pistachios, plums, plums and prunes, pomegranates, potatoes, proso millet, prunes, pumpkins radishes, ra peseed, raspberries

- all, raspberries

- black, raspberries

- red, rhubarb, rice, rice

- wild, rye for grain, safflower, small grain hay, sod harvested (in the open), sorghum for grain, sorghum for silage or greenchop, sorghum for syrup, soursop, soybeans, spinach , squash

- all, squash

- summer, squash

- winter, starfruit, strawberries, sugarbeets for seed, sugarbeets for sugar, sunflower seed

- all, sunflower seed

- non

- oil varieties, sunflower seed

- oil varieties, sweet potatoes, tangerines, tobacco, tomatoes in the open, t riticale, turnip greens, turnips, walnuts

- english, watercress, watermelons,

wheat

- all, wheat

- durum, wheatgrass seed, wheat

- spring (other), wheat

- winter, yam (lesser), yams

Common Name Scientific Name Medium STATE NAME County Taxon
Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Alabama Bibb Insect
Snail, Tulotoma Tulotoma magnifica Terrestrial Alabama Clay Gastropod
Snail, Tulotoma Tulotoma magnifica Terrestrial Alabama Coosa Gastropod
Snail, Tulotoma Tulotoma magnifica Terrestrial Alabama Elmore Gastropod Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Alabama Greene Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Alabama Jackson Insect
Snail, Tulotoma Tulotoma magnifica Terrestrial Alabama Monroe Gastropod
Snail, Tulotoma Tulotoma magnifica Terrestrial Alabama Shelby Gastropod
Snail, Tulotoma Tulotoma magnifica Terrestrial Alabama St. Clair Gastropod
Snail, Tulotoma Tulotoma magnifica Terrestrial Alabama Talladega Gastropod Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Alabama Tuscaloosa Insect Springsnail, San

Bernardino Pyrgulopsis

bernardina Terrestrial,

Freshwater Arizona Cochise Gastropod

Ambersnail, Kanab Oxyloma haydeni

kanabensis Terrestrial,

Freshwater Arizona Coconino Gastropod

Beetle, American Burying Nicrophorus

americanus Terrestrial Arkansas Franklin Insect18 Beetle, American Burying Nicrophorus

americanus Terrestrial Arkansas Little River Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Arkansas Logan Insect Shagreen, Magazine

Mountain Mesodon magazinensis Terrestrial Arkansas Logan Gastropod
Beetle, American Burying Nicrophorus

americanus Terrestrial Arkansas Scott Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Arkansas Sebastian Insect Butterfly, Bay Checkerspot (Wright's euphydryas) Euphydryas editha bayensis Terrestrial California Alameda Insect Butterfly, Callippe Silverspot Speyeria callippe callippe Terrestrial California Alameda Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Amador Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Butte Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Calaveras Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Colusa Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Contra Costa Insect Butterfly, Callippe Silverspot Speyeria callippe callippe Terrestrial California Contra Costa Insect Butterfly, Lange's

Metalmark Apodemia mormo langei Terrestrial California Contra Costa Insect Butterfly, Oregon Silverspot Speyeria zerene hippolyta Terrestrial California Del Norte Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California El Dorado Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Fresno Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Glenn Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Kern Insect
Moth, Kern Primrose Sphinx Euproserpinus

euterpe Terrestrial California Kern Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Kings Insect Beetle, Valley Desmocerus Terrestrial California Lake Insect19 Elderberry Longhorn californicus dimorphus Skipper, Carson Wandering Pseudocopaeodes

eunus obscurus Terrestrial California Lassen Insect Butterfly, El Segundo Blue Euphilotes battoides allyni Terrestrial California Los Angeles Insect

Butterfly, Palos

Verdes Blue Glaucopsyche lygdamus palosverdesensis Terrestrial California Los Angeles Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Madera Insect Butterfly, Mission

Blue Icaricia icarioides

missionensis Terrestrial California Marin Insect Butterfly, Myrtle's Silverspot Speyeria zerene myrtleae Terrestrial California Marin Insect Butterfly, San Bruno

Elfin Callophrys mossii bayensis Terrestrial California Marin Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Mariposa Insect Butterfly, Behren's Silverspot Speyeria zerene

behrensii Terrestrial California Mendocino Insect

Butterfly, Lotis Blue Lycaeides argyrognomon lotis Terrestrial California Mendocino Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Merced Insect Butterfly, Smith's

Blue Euphilotes enoptes

smithi Terrestrial California Monterey Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Napa Insect Butterfly, Callippe Silverspot Speyeria callippe callippe Terrestrial California Napa Insect Butterfly, Myrtle's Silverspot Speyeria zerene myrtleae Terrestrial California Napa Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Nevada Insect

Butterfly, Quino Checkerspot Euphydryas editha quino (=E. e. wrighti) Terrestrial California Orange Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Placer Insect Beetle, Casey's June Dinacoma caseyi Terrestrial California Riverside Insect

Butterfly, Quino Checkerspot Euphydryas editha quino (=E. e. wrighti) Terrestrial California Riverside Insect

Fly, Delhi Sands

Flower

- loving Rhaphiomidas

terminatus

abdominalis Terrestrial California Riverside Insect

Beetle, Delta Green

Ground Elaphrus viridis Terrestrial,

Vernal pool California Sacramento Insect20 Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Sacramento Insect

Butterfly, Quino Checkerspot Euphydryas editha quino (=E. e. wrighti) Terrestrial California San Bernardino Insect

Fly, Delhi Sands

Flower

- loving Rhaphiomidas

terminatus

abdominalis Terrestrial California San Bernardino Insect

Butterfly, Quino Checkerspot Euphydryas editha quino (=E. e. wrighti) Terrestrial California San Diego Insect Skipper, Laguna

Mountain Pyrgus ruralis lagunae Terrestrial California San Diego Insect Butterfly, Mission

Blue Icaricia icarioides

missionensis Terrestrial California San Francisco Insect Butterfly, San Bruno

Elfin Callophrys mossii bayensis Terrestrial California San Francisco Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California San Joaquin Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California San Luis Obispo Insect Butterfly, Smith's

Blue Euphilotes enoptes

smithi Terrestrial California San Luis Obispo Insect
Snail, Morro

Shoulderband Helminthoglypta

walkeriana Terrestrial California San Luis Obispo Gastropod Butterfly, Bay Checkerspot (Wright's euphydryas) Euphydryas editha bayensis Terrestrial California San Mateo Insect Butterfly, Callippe Silverspot Speyeria callippe callippe Terrestrial California San Mateo Insect Butterfly, Mission

Blue Icaricia icarioides

missionensis Terrestrial California San Mateo Insect Butterfly, Myrtle's Silverspot Speyeria zerene myrtleae Terrestrial California San Mateo Insect Butterfly, San Bruno

Elfin Callophrys mossii bayensis Terrestrial California San Mateo Insect Butterfly, Bay Checkerspot (Wright's euphydryas) Euphydryas editha bayensis Terrestrial California Santa Clara Insect
Beetle, Mount

Hermon June Polyphylla barbata Terrestrial,

Subterraneous California Santa Cruz Insect Beetle, Ohlone Tiger Cicindela ohlone Terrestrial California Santa Cruz Insect Grasshopper, Zayante Band

winged Trimerotropis infantilis Terrestrial California Santa Cruz Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Shasta Insect
Beetle, Delta Green

Ground Elaphrus viridis Terrestrial, Vernal pool California Solano Insect21 Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Solano Insect Butterfly, Callippe Silverspot Speyeria callippe callippe Terrestrial California Solano Insect Butterfly, Behren's Silverspot Speyeria zerene

behrensii Terrestrial California Sonoma Insect Butterfly, Myrtle's Silverspot Speyeria zerene myrtleae Terrestrial California Sonoma Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Stanislaus Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Sutter Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Tehama Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Tulare Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Tuolumne Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Yolo Insect

Beetle, Valley Elderberry Longhorn Desmocerus californicus dimorphus Terrestrial California Yuba Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Chaffee Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Dolores Insect Skipper, Pawnee

Montane Hesperia leonardus

montana Terrestrial Colorado Douglas Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Eagle Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Gunnison Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Hinsdale Insect Skipper, Pawnee

Montane Hesperia leonardus

montana Terrestrial Colorado Jefferson Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado La Plata Insect Butterfly, Uncompahgre Boloria acrocnema Terrestrial Colorado Lake Insect Fritillary22 Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Mineral Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Ouray Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Park Insect Skipper, Pawnee

Montane Hesperia leonardus

montana Terrestrial Colorado Park Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Pitkin Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Rio Grande Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Saguache Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado San Miguel Insect Butterfly, Uncompahgre Fritillary Boloria acrocnema Terrestrial Colorado Summit Insect Skipper, Pawnee

Montane Hesperia leonardus

montana Terrestrial Colorado Teller Insect

Beetle, Puritan Tiger Cicindela puritana Terrestrial,

Coastal Connecticut Hartford Insect

Beetle, Puritan Tiger Cicindela puritana Terrestrial,

Coastal Connecticut Middlesex Insect Butterfly [Cassius Blue, Ceraunus Blue, Nickerbean Blue] Leptotes and Hemiargus and Cyclargus genus Terrestrial Florida Miami

- Dade Insect Butterfly, Ceranus

Blue Hemiargus ceraunus

antibubastus Terrestrial Florida Miami

- Dade Insect Butterfly, Miami

Blue Cyclargus thomasi

bethunebakeri Terrestrial Florida Miami

- Dade Insect Butterfly, Nickerbean

Blue Cyclargus ammon Terrestrial Florida Miami

- Dade Insect Butterfly [Cassius Blue, Ceraunus Blue, Nickerbean Blue] Leptotes and Hemiargus and Cyclargus genus Terrestrial Florida Monroe Insect Butterfly, Ceranus

Blue Hemiargus ceraunus

antibubastus Terrestrial Florida Monroe Insect Butterfly, Miami

Blue Cyclargus thomasi

bethunebakeri Terrestrial Florida Monroe Insect Butterfly, Nickerbean

Blue Cyclargus ammon Terrestrial Florida Monroe Insect

Butterfly, Schaus

Swallowtail Heraclides

aristodemus

ponceanus Terrestrial Florida Monroe Insect
Snail, Stock Island

Tree Orthalicus reses (not incl. nesodryas) Terrestrial Florida Monroe Gastropod23 Damselfly, Pacific

Hawaiian Megalagrion pacificum Terrestrial Hawaii Hawaii Insect Fly, Hawaiian picture

- wing Drosophila

heteroneura Terrestrial Hawaii Hawaii Insect Fly, Hawaiian picture

- wing Drosophila mulli Terrestrial Hawaii Hawaii Insect Fly, Hawaiian picture

- wing Drosophila

ochrobasis Terrestrial Hawaii Hawaii Insect
Moth, Blackburn's Sphinx Manduca blackburni Terrestrial Hawaii Hawaii Insect Damselfly, Pacific

Hawaiian Megalagrion pacificum Terrestrial Hawaii Kauai Insect

Fly, Hawaiian picture

- wing Drosophila musaphilia Terrestrial Hawaii Kauai Insect Hawaiian picture

- wing Fly Drosophila sharpi Terrestrial Hawaii Kauai Insect
Moth, Blackburn's Sphinx Manduca blackburni Terrestrial Hawaii Kauai Insect Spider, Kauai Cave

Wolf Adelocosa anops Terrestrial,

Subterraneous Hawaii Kauai Arachnid Damselfly, Flying Earwig Hawaiian Megalagrion

nesiotes Terrestrial Hawaii Maui Insect Damselfly, Pacific

Hawaiian Megalagrion pacificum Terrestrial Hawaii Maui Insect Fly, Hawaiian picture

- wing Drosophila

neoclavisetae Terrestrial Hawaii Maui Insect
Moth, Blackburn's Sphinx Manduca blackburni Terrestrial Hawaii Maui Insect

blackline Hawaiian damselfly Megalagrion nigrohamatum nigrolineatum Terrestrial Hawaii Oahu Insect

Crimson Hawaiian damselfly Megalagrion leptodemas Terrestrial Hawaii Oahu Insect Fly, Hawaiian picture

- wing Drosophila aglaia Terrestrial Hawaii Oahu Insect Fly, Hawaiian picture

- wing Drosophila montgomeryi Terrestrial Hawaii Oahu Insect Fly, Hawaiian picture

- wing Drosophila obatai Terrestrial Hawaii Oahu Insect Fly, Hawaiian picture

- wing Drosophila substenoptera Terrestrial Hawaii Oahu Insect Fly, Hawaiian picture

- wing Drosophila tarphytrichia Terrestrial Hawaii Oahu Insect Fly, Hawaiian picture

- wing Drosophila hemipeza Terrestrial Hawaii Oahu Insect
Moth, Blackburn's Sphinx Manduca blackburni Terrestrial Hawaii Oahu Insect

Oceanic Hawaiian damselfly Megalagrion

oceanicum Terrestrial Hawaii Oahu Insect
Snail, O'ahu Tree (Achatinella abbreviata) Achatinella

abbreviata Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella Achatinella apexfulva Terrestrial Hawaii Oahu Gastropod24 apexfulva)
Snail, O'ahu Tree (Achatinella bellula) Achatinella bellula Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella buddii) Achatinella buddii Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella bulimoides) Achatinella

bulimoides Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella byronii) Achatinella byronii Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella caesia) Achatinella caesia Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella casta) Achatinella casta Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella cestus) Achatinella cestus Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella concavospira) Achatinella concavospira Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella curta) Achatinella curta Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella decipiens) Achatinella decipiens Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella decora) Achatinella decora Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella dimorpha) Achatinella dimorpha Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella elegans) Achatinella elegans Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella fulgens) Achatinella fulgens Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella fuscobasis) Achatinella fuscobasis Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella juddii) Achatinella juddii Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella juncea) Achatinella juncea Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella lehuiensis) Achatinella

lehuiensis Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella leucorraphe) Achatinella leucorraphe Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella lila) Achatinella lila Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella livida) Achatinella livida Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella lorata) Achatinella lorata Terrestrial Hawaii Oahu Gastropod

Snail, O'ahu Tree (Achatinella mustelina) Achatinella

mustelina Terrestrial Hawaii Oahu Gastropod25 Snail, O'ahu Tree (Achatinella papyracea) Achatinella

papyracea Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella phaeozona) Achatinella phaeozona Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella pulcherrima) Achatinella pulcherrima Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella pupukanioe) Achatinella pupukanioe Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella rosea) Achatinella rosea Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella sowerbyana) Achatinella sowerbyana Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella spaldingi) Achatinella spaldingi Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella stewartii) Achatinella stewartii Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella swiftii) Achatinella swiftii Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella taeniolata) Achatinella

taeniolata Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella thaanumi) Achatinella

thaahumi Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella turgida) Achatinella turgida Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella valida) Achatinella valida Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella viridans) Achatinella viridans Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella vittata) Achatinella vittata Terrestrial Hawaii Oahu Gastropod
Snail, O'ahu Tree (Achatinella vulpina) Achatinella vulpina Terrestrial Hawaii Oahu Gastropod
Snail, Snake River Physa Physa natricina Terrestrial Idaho Cassia Gastropod
Snail, Snake River Physa Physa natricina Terrestrial Idaho Elmore Gastropod
Snail, Snake River Physa Physa natricina Terrestrial Idaho Gooding Gastropod
Snail, Snake River Physa Physa natricina Terrestrial Idaho Jerome Gastropod
Snail, Snake River Physa Physa natricina Terrestrial Idaho Minidoka Gastropod
Snail, Snake River Physa Physa natricina Terrestrial Idaho Owyhee Gastropod
Snail, Snake River Physa natricina Terrestrial Idaho Twin Falls Gastropod26 Physa Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Illinois Cook Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Illinois Du Page Insect
Snail, Iowa

Pleistocene Discus macclintocki Terrestrial Illinois Jo Daviess Gastropod Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Illinois Lake Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Illinois Will Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Indiana La Porte Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Indiana Lagrange Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Indiana Lake Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Indiana Porter Insect
Snail, Iowa

Pleistocene Discus macclintocki Terrestrial Iowa Clayton Gastropod
Snail, Iowa

Pleistocene Discus macclintocki Terrestrial Iowa Clinton Gastropod
Snail, Iowa

Pleistocene Discus macclintocki Terrestrial Iowa Dubuque Gastropod
Snail, Iowa

Pleistocene Discus macclintocki Terrestrial Iowa Fayette Gastropod
Snail, Iowa

Pleistocene Discus macclintocki Terrestrial Iowa Jackson Gastropod
Beetle, American Burying Nicrophorus

americanus Terrestrial Kansas Chautauqua Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Kansas Elk Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Kansas Montgomery Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Kansas Wilson Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Kentucky Henderson Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Kentucky Jefferson Insect Beetle, American Burying Nicrophorus

americanus Terrestrial Kentucky Trigg Insect
Beetle, Northeastern Beach Tiger Cicindela dorsalis

dorsalis Terrestrial Maryland Calvert Insect

Beetle, Puritan Tiger Cicindela puritana Terrestrial,

Coastal Maryland Calvert Insect

Beetle, Puritan Tiger Cicindela puritana Terrestrial,

Coastal Maryland Cecil Insect

Beetle, Puritan Tiger Cicindela puritana Terrestrial,

Coastal Maryland Kent Insect
Beetle, Northeastern Beach Tiger Cicindela dorsalis

dorsalis Terrestrial Maryland Somerset Insect
Beetle, Northeastern Beach Tiger Cicindela dorsalis

dorsalis Terrestrial Massachusetts Barnstable Insect27 Beetle, American Burying Nicrophorus

americanus Terrestrial Massachusetts Dukes Insect
Beetle, Northeastern Beach Tiger Cicindela dorsalis

dorsalis Terrestrial Massachusetts Dukes Insect

Beetle, Puritan Tiger Cicindela puritana Terrestrial,

Coastal Massachusetts Hampshire Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Massachusetts Nantucket Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Michigan Allegan Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Michigan Alpena Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Michigan Barry Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Michigan Berrien Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Michigan Branch Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Michigan Cass Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Michigan Ionia Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Michigan Jackson Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Michigan Kalamazoo Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Michigan Kent Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Michigan Lake Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Michigan Mackinac Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Michigan Mason Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Michigan Mecosta Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Michigan Monroe Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Michigan Montcalm Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Michigan Muskegon Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Michigan Newaygo Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Michigan Oceana Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Michigan Presque Isle Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Michigan St. Joseph Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Michigan Van Buren Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Michigan Washtenaw Insect Butterfly, Karner Lycaeides melissa Terrestrial Minnesota Winona Insect28 Blue samuelis Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Missouri Dent Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Missouri Iron Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Missouri Phelps Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Missouri Reynolds Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Missouri Ripley Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Missouri Shannon Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Missouri Wayne Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Blaine Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Boyd Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Brown Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Cherry Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Custer Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Dawson Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Frontier Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Garfield Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Gosper Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Holt Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Keya Paha Insect
Beetle, Salt Creek Tiger Cicindela nevadica

lincolniana Terrestrial Nebraska Lancaster Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Lincoln Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Loup Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Rock Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Thomas Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Nebraska Wheeler Insect Skipper, Carson Wandering Pseudocopaeodes

eunus obscurus Terrestrial Nevada Carson City Insect
Naucorid, Ash

Meadows Ambrysus

amargosus Terrestrial Nevada Nye Insect Skipper, Carson Wandering Pseudocopaeodes

eunus obscurus Terrestrial Nevada Washoe Insect29 Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial New Hampshire Merrimack Insect
Beetle, Northeastern Beach Tiger Cicindela dorsalis

dorsalis Terrestrial New Jersey Monmouth Insect Springsnail, Koster's Juturnia kosteri Terrestrial New Mexico Chaves Gastropod Springsnail, Chupadera Pyrgulopsis chupaderae Terrestrial,

Freshwater New Mexico Socorro Gastropod Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial New York Albany Insect Snail, Chittenango

Ovate Amber Succinea chittenangoensis Terrestrial,

Freshwater New York Madison Gastropod Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial New York Saratoga Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial New York Schenectady Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial New York Warren Insect Spider, Spruce

- fir

Moss Microhexura montivaga Terrestrial North Carolina Avery Arachnid Spider, Spruce

- fir

Moss Microhexura montivaga Terrestrial North Carolina Caldwell Arachnid Butterfly, Saint Francis' Satyr Neonympha mitchellii francisci Terrestrial North Carolina Craven Insect Spider, Spruce

- fir

Moss Microhexura montivaga Terrestrial North Carolina Haywood Arachnid Butterfly, Saint Francis' Satyr Neonympha mitchellii francisci Terrestrial North Carolina Hoke Insect Spider, Spruce

- fir Microhexura Terrestrial North Carolina Mitchell Arachnid

Moss montivaga

Snail, Noonday Mesodon clarki

nantahala Terrestrial North Carolina Swain Gastropod Spider, Spruce

- fir

Moss Microhexura montivaga Terrestrial North Carolina Swain Arachnid Spider, Spruce

- fir

Moss Microhexura montivaga Terrestrial North Carolina Watauga Arachnid Spider, Spruce

- fir

Moss Microhexura montivaga Terrestrial North Carolina Yancey Arachnid
Beetle, American Burying Nicrophorus

americanus Terrestrial Ohio Athens Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Ohio Hocking Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Ohio Lucas Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Ohio Portage Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Ohio Vinton Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Adair Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Atoka Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Bryan Insect
Beetle, American Nicrophorus Terrestrial Oklahoma Cherokee Insect30 Burying americanus Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Choctaw Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Coal Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Craig Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Creek Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Delaware Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Haskell Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Hughes Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Johnston Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Latimer Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Le Flore Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Marshall Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Mayes Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma McCurtain Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma McIntosh Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Muskogee Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Nowata Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Okfuskee Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Okmulgee Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Osage Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Ottawa Insect Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Pawnee Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Pittsburg Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Pontotoc Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Pushmataha Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Rogers Insect

Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Seminole Insect
Beetle, American Nicrophorus Terrestrial Oklahoma Sequoyah Insect31 Burying americanus Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Tulsa Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Wagoner Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Oklahoma Washington Insect Butterfly, Fender's

Blue Icaricia icarioides fenderi Terrestrial Oregon Benton Insect Butterfly, Oregon Silverspot Speyeria zerene hippolyta Terrestrial Oregon Clatsop Insect Butterfly, Fender's

Blue Icaricia icarioides fenderi Terrestrial Oregon Lane Insect Butterfly, Oregon Silverspot Speyeria zerene hippolyta Terrestrial Oregon Lane Insect Butterfly, Oregon Silverspot Speyeria zerene hippolyta Terrestrial Oregon Lincoln Insect Butterfly, Fender's

Blue Icaricia icarioides fenderi Terrestrial Oregon Linn Insect Butterfly, Fender's

Blue Icaricia icarioides fenderi Terrestrial Oregon Polk Insect Butterfly, Oregon Silverspot Speyeria zerene hippolyta Terrestrial Oregon Tillamook Insect Butterfly, Fender's

Blue Icaricia icarioides fenderi Terrestrial Oregon Yamhill Insect Butterfly, Oregon Silverspot Speyeria zerene hippolyta Terrestrial Oregon Yamhill Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Rhode Island Washington Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial South Dakota Bennett Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial South Dakota Brookings Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial South Dakota Gregory Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial South Dakota Haakon Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial South Dakota Todd Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial South Dakota Tripp Insect Beetle, American Burying Nicrophorus

americanus Terrestrial South Dakota Union Insect Spider, Spruce

- fir

Moss Microhexura montivaga Terrestrial Tennessee Carter Arachnid
Snail, Painted Snake

Coiled Forest Anguispira picta Terrestrial Tennessee Franklin Gastropod Marstonia, Royal (=Royal Snail) Pyrgulopsis ogmorhaphe Terrestrial Tennessee Marion Gastropod Spider, Spruce

- fir

Moss Microhexura montivaga Terrestrial Tennessee Sevier Arachnid

Beetle, Helotes Mold Batrisodes venyivi Subterraneous Texas Bexar Insect
Harvestman, Cokendolpher Cave Texella cokendolpheri Terrestrial,

Subterraneous Texas Bexar Arachnid32 Meshweaver, Braken

Bat Cave Cicurina venii Terrestrial,

Subterraneous Texas Bexar Arachnid
Meshweaver, Government Canyon

Bat Cave Cicurina vespera Terrestrial,

Subterraneous Texas Bexar Arachnid
Meshweaver,

Madla's Cave Cicurina madla Terrestrial,

Subterraneous Texas Bexar Arachnid
Meshweaver, Robber

Baron Cave Cicurina baronia Terrestrial,

Subterraneous Texas Bexar Arachnid

Rhadine exilis (ncn) Rhadine exilis Terrestrial,

Subterraneous Texas Bexar Insect

Rhadine infernalis (ncn) Rhadine infernalis Terrestrial,

Subterraneous Texas Bexar Insect Spider, Government Canyon Bat Cave Neoleptoneta microps Terrestrial,

Subterraneous Texas Bexar Arachnid
Harvestman, Bee

Creek Cave Texella reddelli Terrestrial,

Subterraneous Texas Burnet Arachnid
Beetle, American Burying Nicrophorus

americanus Terrestrial Texas Lamar Insect
Beetle, American Burying Nicrophorus

americanus Terrestrial Texas Red River Insect
Beetle, Kretschmarr

Cave Mold Texamaurops

reddelli Subterraneous Texas Travis Insect
Beetle, Tooth Cave

Ground Rhadine persephone Subterraneous Texas Travis Insect
Harvestman, Bee

Creek Cave Texella reddelli Terrestrial,

Subterraneous Texas Travis Arachnid
Harvestman, Bone

Cave Texella reyesi Terrestrial,

Subterraneous Texas Travis Arachnid Pseudoscorpion,

Tooth Cave Tartarocreagris

texana Terrestrial,

Subterraneous Texas Travis Arachnid

Spider, Tooth Cave Leptoneta myopica Terrestrial,

Subterraneous Texas Travis Arachnid
Beetle, Coffin Cave

Mold Batrisodes texanus Subterraneous Texas Williamson Insect
Beetle, Tooth Cave

Ground Rhadine persephone Subterraneous Texas Williamson Insect
Harvestman, Bone

Cave Texella reyesi Terrestrial,

Subterraneous Texas Williamson Arachnid

Ambersnail, Kanab Oxyloma haydeni

kanabensis Terrestrial,

Freshwater Utah Kane Gastropod
Beetle, Northeastern Beach Tiger Cicindela dorsalis

dorsalis Terrestrial Virginia Accomack Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Virginia Carroll Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Virginia Floyd Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Virginia Franklin Insect
Spider, Spruce

- fir

Moss Microhexura montivaga Terrestrial Virginia Grayson Arachnid
Beetle, Northeastern Beach Tiger Cicindela dorsalis

dorsalis Terrestrial Virginia Lancaster Insect
Beetle, Northeastern Cicindela dorsalis Terrestrial Virginia Mathews Insect33 Beach Tiger dorsalis
Beetle, Northeastern Beach Tiger Cicindela dorsalis

dorsalis Terrestrial Virginia Middlesex Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Virginia Montgomery Insect
Beetle, Northeastern Beach Tiger Cicindela dorsalis

dorsalis Terrestrial Virginia Northampton Insect
Beetle, Northeastern Beach Tiger Cicindela dorsalis

dorsalis Terrestrial Virginia Northumberland Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Virginia Patrick Insect Butterfly, Mitchell's Satyr Neonympha

mitchellii mitchellii Terrestrial,

Perm. wetland Virginia Pulaski Insect Snail, Virginia Fringed Mountain Polygyriscus virginianus Terrestrial Virginia Pulaski Gastropod Butterfly, Oregon Silverspot Speyeria zerene hippolyta Terrestrial Washington Grays Harbor Insect Butterfly, Oregon Silverspot Speyeria zerene hippolyta Terrestrial Washington Pacific Insect
Snail, Flat

- spired

Three

- toothed Triodopsis platysayoides Terrestrial West Virginia Monongalia Gastropod
Snail, Flat

- spired

Three

- toothed Triodopsis platysayoides Terrestrial West Virginia Preston Gastropod Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Adams Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Barron Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Burnett Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Chippewa Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Clark Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Wisconsin Door Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Dunn Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Eau Claire Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Green Lake Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Jackson Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Juneau Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Kenosha Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Wisconsin Kewaunee Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Marquette Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Menominee Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Monroe Insect34 Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Oconto Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Outagamie Insect Dragonfly, Hine's

Emerald Somatochlora

hineana Terrestrial,

Freshwater Wisconsin Ozaukee Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Polk Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Portage Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Shawano Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin St. Croix Insect
Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Waupaca Insect Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Waushara Insect

Butterfly, Karner

Blue Lycaeides melissa

samuelis Terrestrial Wisconsin Wood Insect