~

NOAA Technical Memorandum

NMFS-SEFC-158

The Husbandry of Hatchling

to Yearling Kemp's Ridley

Sea Turtles (Lepidochelys Kempi)

JULY 1985

u.s. DEPARTMENT OF COMMERCE

National Oceanic and Atmospheric Administration·

National Marine Fisheries Service

Southeast Fisheries Center

Galveston Laboratory

Galveston, Texas 77550
TABLE OF CONTENTS

Page

NOTI CE •••••••••••••••••••••••••••••••••••••••••••.•••••••••••••• i

FOREWORD ••••••••••••••••••••••••••••••••••••••••••••••••••••••• ii

INTRODUCTION •••••••••••••••••••••••••••••••••••••••••••••••••••

PURPOSE ••••••••• -••••••••••••••••••••••••••••••••••••••••••••••• 5

SEA TURTLE HEAD START RESEARCH FACILITIES •••••••••••••••••••••• 6

Location ••••••.•••••••••••••••••••••••.•••••••••••••••.•••• 6

Seawater •••••••.•.••..•••••••.••••••••••••••.••••••••••••..• 6

FaciIi ties .••......•.....••••...••...•.•••.•..•.•..•....... 7

Equi pmen t - 0 7

Wastewater Treatm~nt•••••••~••••••••••••••••••••••••••••••• 9

HEAD START OPERATIONS •••••••••••••••••••••••••••••••••••••••••• 11

Receipt of Hatchlings •••••••••••••••••••••••••••••••••••••• 11

Weighing and Measuring ••••••••••••••••••••••••••••••••••••• 12

Feeding •••••••••••••••••••••••••••••••••••••••••••••••••••• 13

Growth ••••••••••••••••••••••••••.••••••.••••••••••••• '•••••••• 15

Heal th Care •••••••••••••••••••••••••••••••••••••••••••••••• 15

Tagging •••••••••••••••••••••••••••••• it ••••••••••••••••••••• 16

Transportation to Release Sites •••••••••••••••••••••••••••• 17

Re lease ••••••••••••••• It •••••••••••••••••••••••••••••••••••• 18

SEA TURTLE HEAD START RESEARCH AND GENERAL

OBSERVATIONS •••••••••••••••••••••••••••••••••••••••••••••••••

DISCUSSION •••••••••••••••••••••••••••••••••••••••••••••••••••••

EP ILOGUE •••••••••••••••••••••••••••••••••••••••••••••••••••••••

General Observations.- ••• '.•••.•••••••••••.••••••••••••••.•.•

19

19

20

20

22

23

23

25

26

28

29

32

.........

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Recaptures of Head Started/Tagged/Released Turtles •••••••••

Feeding Regimen and Food Utilization •••••••••••••••••••••••

Food Color Preference •••••••••••.••••••••••• -••••••••••••••••

Re-cycled seawater •••••••••••••••••••••••••••••••••.•••••••

Tagging •.•••••••••••••.•••.•••••••••••••••..•••••••••••••••

Celestial Orientation of Hatchlings ••••••••••••••••••••••••

ACKNOWLEDGMENTS •••••••••••••••••••••••••••••••••••••••

LITERATURE CITED ••••••••••••••••••••••••••••••••••••••
NOTICE

This report should be cited as follows:

Fontaine, C. T., K. T. Marvin, T. D. Williams, W. J. Browning,

R. M. Harris, K. L. W. Indelicato, G. A. Shattuck

and R. A. Sadler. 1985. The husbandry of hatchling

to yearling Kemp's ridley sea turtles (lepidochelys kempi).

NOAA Technical Memorandum NMFS-SEFC-158, iii + 34 p.,

10 Tables, 22 Figures, and 2 Appendices.

i
FOREWORD

Head starting is one of a number of potentially effective conservation and management tools that may help prevent extinction of the

seriously endangered Kemp's ridley sea turtle, Lepidochelys kempi.

However, head starting's utility and effectiveness in conservation

and management are unproven (Klima and McVey, 1981). Though seven

year-classes (1978-1984) of Kemp's ridley have been head started at

the National Marine Fisheries Service, Southeast Fisheries Center's

Galveston Laboratory, and 9,258 survivors have been tagged and

released into the Gulf of Mexico, none of these animals has returned

to nest so far as is known. Therefore, head starting is still experimental.

Head starting is expensive (exceeding $150 per animal per year)

and labor-intensive, requiring considerable resources of personnel,

facilities, and supplies. However, these costs are not high compared

to those of maintaining and breeding other large tetrapods in captivity; e.g., for medical research, to prevent extinction of

endangered species, or for public display.

Because each species of sea turtle has somewhat different and

unique traits that adapt it for survival, each also has somewhat

different requirements for captive rearing. As an example, Kemp's

ridley is aggressive, so its young are reared in isolation to prevent

them from injuring each other.

It is encouraging that two five-year-old, head started Kemp's

ridleys, held in captivity through their entire life, laid eggs at the

Cayman Turtle Farm [1983] Ltd. on Grand Cayman Island in May 1984

(Wood and Wood 1984). In spring 1985 there were additional reports of

reproductive behavior in the Kemp's ridleys at the farm (James Wood,

personal communication). These events document the earliest age for

nesting in the species. Though none of the hatchlings survived, such

early nesting is an encouraging initial step toward captive breeding

as a "safety-net" for the species. It also adds impetus to the Kemp's

ridley head start research project by giving an indication that head

started, tagged ridleys released into the Gulf of Mexico may approach

maturity more rapidly than was originally thought possible.

ii
In this paper on the husbandry of Kemp's ridley sea turtle, the

authors describe the facilities, equipment, methods and techniques

adapted and developed by the Galveston Laboratory. These methods not

only have provided the animals tagged and released into the gulf, but

also 210 yearlings that have been distributed among eight organizations for extended head starting or captive breeding. These organizations are now maintaining 75 survivors. Some of the survivors may be

released at a later date, and others retained in captivity as a potential brood stock. Plans are to add up to 50 more ridleys of the 1984

year-class to this captive stock. Of the turtles that were tagged and

released, 380 have been recovered to date (Sharon Manzella, personal

communication, June 1985), providing information on tag retention,

survival, growth and dispersion within the Gulf of Mexico, along the

eastern coast of the United States, and in European Atlantic waters.

According to Dr. Edward F. Klima, Galveston Laboratory Director,

Southeast Fisheries Center, Galveston, Texas, those who use this paper

should be cautioned that experimental head starting requires a longterm commitment (10-15 years). Successful rearing of sea turtles in

captivity requires considerable dedication among the personnel

involved and substantial financial resources as well as an appropriate

infrastructure, both within and outside any organization or agency

that chooses to investigate head starting of sea turtles as a potential management tool. Ultimate success of head starting can be

achieved only with the nesting of head started turtles and survival of

their young. That will require their survival, growth, maturation, and

successful migration to a nesting beach as well as successful mating,

nesting and hatching. Many questions remain concerning captive

rearing. For example, most of the head started turtles have been

released as yearlings, but younger or older animals might be better

for release.

This paper contains information supplementing currently available

literature on the rearing and maintenance of young sea turtles in captivity. On the short-term, much has been learned by successful head

starting of Kemp's ridley, but greater public awareness and support

are required to generate the additional resources needed to continue

iii
and to expand the project as well as test its success as one means of

saving the Kemp's ridley from extinction.

Charles W. Caillouet, Jr., Ph.D.

Chief, Life Studies Division

NMFS Galveston Laboratory

June 1985

iv
INTRODUC'l'ION

The Kemp's ridley sea turtle Lepidochelys kempi, has been on the

endangered species list since the passage of the u.s. Endangered

Species Act in 1973. The Kemp's ridley nests each year on a beach

bordering the Gulf of Mexico near the village of Rancho Nuevo, in the

state of Tamaulipas, Mexico (c~vez et al. 1968). In 1947, Hildebrand

(1963) estimated that 40,000 female Kemp's ridleys were nesting on

this beach, but by 1982 the number had declined to about 1,500

(M(arquez et al. 1981). The primary cause of the population decline is

thought to be overexploitation of the adults and eggs by man.

Presently, the nesting beach near Rancho Nuevo is protected during the

nesting season from April to July by Mexican Marines and personnel of

the Instituto Nacional de la Pesca (INP) of Mexico, with assistance

from the u.s. Fish and wildlife Service (FWS) and others. This protection has reduced the poaching by man (~rquez et al. 1982).

Since 1978, the National Marine Fisheries Service (NMFS),

Southeast Fisheries Center (SEFC), Galveston Laboratory has participated in the international program to save the endangered Kemp's

ridley sea turtle from extinction (Klima and McVey 1981). The program

is a joint conservation effort among the INP, the FWS, the NMFS, the

National Park Service (NPS), and the Texas Parks and wildlife Department (TPWD), with assistance from the u.S. Navy and Coast Guard, and

the Florida Department of Natural Resources (FDNR).

The goal of this program is to increase the Kemp's ridley population. The approach includes protection of nesting turtles and their

eggs on the beach at Rancho Nuevo, prohibitions on the capture,

possession and sale of sea turtles in the united States, use of the

trawling efficiency device (TED) to allow escapement of turtles captured incidentally in shrimp trawls, and head starting Kemp's ridleys

in captivity during their first year of life. Head started Kemp's

ridleys, "imprinted" as hatchlings at Padre Island, have been tagged

and released in hopes of establishing a new nesting colony at the
Padre Island National Seashore, near Corpus Christi, Texas. Others

have been "imprinted" at Rancho Nuevo to supplement that breeding

colony.

Head starting involves the husbandry of young turtles during the

critical first year of life as well as the tagging and release of survivors. In addition, the program has provided answers to many

questions concerning biology, physiology, and pathology of this

endangered species, as well as the fate of cultured turtles at sea.

The sea turtle head start research project at the NMFS Galveston

Laboratory falls within the Protected Species Program administered by

the NMFS SEFC, in Miami, Florida. Project objectives are to:

(1) provide husbandry to captive Kemp's ridley sea turtles to

increase their survival and optimize their growth during the

critical first year of life;

(2) tag healthy survivors of each year-class of head started

Kemp's ridley sea turtles and release them into their natural

environment;

(3) monitor growth, survival and migration of head started Kemp's

ridleys based on reported recoveries;

(4) establish a new nesting colony of Kemp's ridleys at the Padre

Island National Seashore; and

(5) develop and expand a captive brood stock.

Criteria for gauging success in the project are:

(1) survival and growth of head started animals in captivity

during the first year of life;

2
(2) nesting of head started animals on any beach;

(3) nesting of head started animals on beaches to which they

were exposed as eggs and hatchlings; and

(4) copulation and nesting within the captive brood stock of

head started animals.

In theory, imprinting (species-specific, rapid learning during a

critical time of early life in which social attachment and identification are established) occurs during incubation and hatching of

the eggs in beach sand and during exposure of hatchlings to the adjacent surf, thus acting as a memory code to guide adult turtles back to

their natal beach. At Rancho Nuevo each season, a portion of the eggs

are collected in plastic bags as they are laid so they do not touch

the local beach sand. These eggs are placed in polystyrene foam boxes

containing beach sand from Padre Island and are flown to the Padre

Island National Seashore where they are placed in a hatchery and

allowed to continue incubation. NPS biologists carefully monitor

environmental conditions in the boxes during the incubation that normally takes between 43 and 53 days (Robert King, NPS, personal communication, July 1984). Hatchlings are taken to the beach and allowed

to crawl across the sand to the surf to enhance their opportunity for

imprinting. The hatchlings are recovered from the surf, placed in

boxes lined with polyurethane foam cushions and flown by u.S. Navy

aircraft to Galveston. In the past, some eggs also have been

collected, incubated, and hatched in Rancho Nuevo beach sand and the

hatchlings reared at Galveston.

Head starting, as practiced by the NMFS Galveston Laboratory,

is thought to give the young Kemp's ridleys an advantage in survival

and growth over their natural counterparts (Klima and McVey 1981;

3
Fontaine, Leong and Caillouet 1983; Caillouet 1984; Caillouet et

al.1/; Fontaine et al.l!). We believe that natural survival of this

turtle during its critical first year of life is less than 1%.

Survival during head starting over a 7-12 month period in captivity

has ranged from 65% to 95% (Table 1). Of the total of 10,376 hatchlings received alive from year-classes 1978-1983, 8,241 (79%) were

reared, tagged and released. Of those released, 378 have been recovered. Growth for each year-class, as expressed by plots of arithmetic average weight versus time, is presented in Fig. 1. Caillouet

et al.1/ examined growth and survival of the 1978-1983 year-classes of

head started Kemp's ridleys in detail.

Most head started and tagged turtles were released into the Gulf

of Mexico, but some were held back as a potential brood stock (Table

1). Twenty-nine of the 1978 year-class, and 50 of the 1979 year-class

were transferred to the Miami Seaquarium (8 survivors of the 1978 and

1 survivor of the 1979 year-classes), Miami, Florida and ten of the

1978 year-class to Sea-Arama Marineworld (8 survivors), Galveston,

Texas. Of the 100 animals from the 1979 year-class that were shipped

to Cancun, Mexico, 96 survivors were transferred to the Cayman Turtle

Farm [1983], Ltd. (34 survivors), on Grand Cayman Island. To each of

the following public marine aquaria (Table 2), 5 turtles of the 1982

year-class were transferred: Clearwater Marine Science Center,

1/caillouet, C. W., Jr., D. B. Koi, C. T. Fontaine, T. D. Williams,

W. J. Browning and R. M. Harris. Growth and survival of Kemp's

ridley sea turtle, Lepidochelys kempi, in captivity. Manuscript

submitted to the editor of Chelonian Documentation Center,

Bulletin.

l!Fontaine, C. T., R. M. Harris, W. J. Browning, and T. D. Williams.

Observations on the growth and distribution of head started/

tagged/released Kemp's ridley sea turtles (Lepidochelys kempi)

from year-classes 1978-1983. Manuscript in preparation. NMFS

SEFC Galveston Laboratory, Galveston, Texas.

4
Clearwater, Florida; Gulfarium, Ft. Walton Beach, Florida; Key West

Municipal Aquarium, Key West, Florida; Turtle Kraals, Key West,

Florida (these were transferred to Theater of the Sea, Islamorada,

Florida on 16 April 1985); and Marine Life, Inc., Gulfport,

Mississippi. All except one turtle that died at Gulfarium remain

alive. Pu~poses of holding these turtles captive beyond one year were

(1) to extend the head starting period for 2-4 additional years thus

providing larger and older animals for release, and (2) to provide a

stock of potential breeders in captivity as a "safety net" for the

species (Caillouet 1984). Some of these captive turtles have been

tagged with the "living tag." The living tag refers to the process

by which a piece of the lighter colored plastron is grafted into the

darker tissue of the carapace. If the tissue graft is successful, a

very distinguishable white mark is made in the darker carapace.

Others have been tagged with binary-coded, internal tags. All were

tagged with monel flipper tags. Thus, these turtles provide an opportunity for determining tag retention and recognition over time. Other

head started Kemp's ridleys having abnormalities, serious injuries, or

incurable diseases were either transferred to universities for

research purposes or humanely disposed of as allowed under FWS permit

(Table 1).

The Kemp's ridley sea turtle head start research project in

Galveston has been conducted under permits issued by the INP, the FWS,

and the TPWD (see ACKNOWLEDGMENTS). Wastewater discharge effluent

quality has been regulated by permit no. 02299 from the Texas

Department of Water Resources (TDWR).

PURPOSE

The purpose of this paper is to document the general husbandry,

including equipment, methods, and techniques, used to rear Kemp's

ridley sea turtles from hatchlings to yearlings. The facilities and

methods have evolved since 1978 based on the cumulative research,

5
experiences and efforts of those engaged in rearing Kemp's ridleys

over the life of the project. This paper primarily presents those

procedures used during the rearing of the 1982-1983 year-classes.

General recommendations for the care and rearing of captive sea

turtles have been presented by Bjorndal and Balazs (1983).

SEA TURTLE HEAD START RESEARCH FACILITIES

Location - The sea turtle head start research facilities are located

at the NMFS SEFC Galveston Laboratory, 4700 Avenue U, Galveston,

Texas (Fig. 2).

Seawater - Seawater is pumped from the Gulf of Mexico through submerged well-points located approximately 200 m from the surfline of

the beach. The seawater is stored initially at the Galveston

Laboratory in underground concrete sump pits (113,460 liter; 30,000

gal) and then pumped to above-ground, redwood storage tanks (94,550

liter; 25,000 gal). Seawater is then delivered from the redwood tanks

to the head start facilities into six, insulated, fiberglass reservoirs (Fig. 3). The four smaller reservoirs have a capacity of

28,390 liter (7,500 gal) each, while the two larger ones have a capacity of 37,850 liter (10,000 gal) each. The two larger reservoirs are

for secondary use to provide back-up supplies of seawater, while the

four smaller reservoirs provide the primary or immediate supply of

seawater. During the months of cold weather, seawater in the four

smaller reservoirs is heated to 280C by thermostatically-controlled

immersion heaters (Table 3). When needed, seawater can either be

drained from any of six reservoirs by gravity-flow or pumped by two

centrifugal pumps (Table 3).

6
Facilities - The sea turtle head start research facilities consist of

two, aluminum-framed, quonset huts (X. S. Smith, Inc.1/) covered by

inflated double-layers of white polyethylene sheathing. Originally,

one quonset hut, 74.4 m (244 ft) long by 9.2 m (30 ft) wide, was

installed. After hurricane Alicia, which inflicted severe damage to

this structure in August 1983 (Fig. 4), two separate quonset huts were

constructed, each 29.3 m (96 ft) long by 9.2 m (30 ft) wide (Fig. 5).

This separation into two parts and the reduction in size of the structure were done to simplify replacement of tops and to reduce the cost

of heating during the winter. The two quonset huts are heated by two,

natural gas-fired, forced-air heaters.

The long axis of the quonset huts is situated on an east-west

orientation so that the prevailing southeasterly winds will provide

cross-ventilation and cooling during the summer. The sides of the

quonset huts are equipped with lateral "vent-rails" located 1.2 m (4

ft) above ground level; panels of polyethylene sheathing attached to

the rails can be removed during summer months for ventilation. The

space between the inflated, double-layered, polyethylene tops is maintained by small fans at approximately 20.3 cm (8 in). This

"air-space" between the layers provides additional insulation. The

tops normally last 12-18 months, but are replaced annually following

the hurricane' season and before winter.

Equipment - The 15 fiberglass raceways (numbered 1-15 from east and

west) and 120 hemispherical basins used to rear turtles in the facility were manufactured by Red Ewald, Inc. The raceways are rectangular (Fig. 6), measuring 6.1 m (20 ft) long x 1.8 m (6 ft) wide x

0.6 m (2 ft) deep, and are normally filled to a depth of 30.5 cm (12

in), providing a seawater volume of 3,141 liter (830 gal). The

raceways are fitted on the north end with a 10.2 cm (4 inch, inside

diameter) standpipe drain (Fig. 7) and a 3.8 cm (1.5 inch, inside

l/Throughout this paper, the mention of trade names or commercial

products does not imply endorsement or recommendation for use.

7
diameter) drain pipe on the bottom at the same end as the standpipe.

The raceways are drained by rotating the 10.2 cm standpipe downward

and removing a rubber stopper from the 3.8 cm bottom drain.

Hatchling Kemp's ridley sea turtles are aggressive and will attack

and injure each other (Klima and McVey 1981; Bjorndal and Balazs

1983); consequently, they cannot be held together in an open tank.

Therefore, each raceway is fitted with 18 5.1 cm (2 in) x 5.1 cm

wooden poles, placed across the width of the raceways (Fig. 8), from

which six plastic buckets, 28 cm (11 in) deep by 22 cm (9 in, inside

diameter), are suspended with 10-gauge galvanized wire. Eighteen such

rows (lettered A-R from south to north) and six columns (numbered 1-6

from east to west) of buckets provide a total of 108 buckets per

raceway. The bottom of each bucket was drilled with 12 holes, each

2.5 cm (1 in) in diameter, to allow water exchange and liberation of

turtle excrement and waste food. Each turtle remains in its assigned

bucket throughout the head start process, unless it dies or becomes

ill (in which case it is transferred to sick bay, treated, and

returned to the same bucket if it is cured). The raceway and bucket

locations provide codes used as identifiers for individual turtles

throughout head starting.

Turtles that out grow their buckets are transferred to fiberglass

basins (Fig. 9). The hemispherical basins are 61 cm (24 in) in

diameter and 25 cm (10 in) deep and are normally filled with 26.5

liter (7 gal) of seawater. The seawater is exchanged and the basins

cleaned on a daily basis.

The sick bay (Fig. 10), where sick turtles are isolated for observation and treatment, contains 86 fiberglass basins similar to those

described in the previous paragraph. When in use, the basins are

drained, scrubbed with a heavy brush, disinfected with Chlorox,

rinsed, and filled with clean seawater on a daily basis.

8
wastewater Treatment - When in use, the raceways are scrubbed once

each week, five (raceways 1-5) on Monday, five (raceways 6-10) on

wednesday, and five (raceways 11-15) on Friday. Cleaning of a raceway involves draining the water, washing down the turtles, buckets,

and raceway with freshwater (tapwater)~ rinsing out the raceway with

freshwater,' scrubbing the inside walls of the raceway with heavy-duty

brushes to remove attached algae, rinsing the tank once again with

freshwater, and refilling the tank with clean seawater. The fresh

tapwater is not heated in the winter time prior to use. Apparently

this has caused no ill effects to the turtles.

The drained seawater and water from scrubbings and rinsings are

collected in a concrete trough (Fig. 7) that empties into a 0.9 m (3

ft) by 1.8 m (6 ft) fiberglass sump pit located outside of the turtle

facility. Two solid-waste sump pumps (Table 2) transfer this water

into two cylindrical, fiberglass, digestion tanks, 1.5 m (5 ft) high

by 6.1 m (20 ft) in diameter (Fig. 11), each holding a volume of

44,568 liter (11,775 gal).

The treatment of the wastewater is based on aerobic digestion of

proteinacious material and was developed with the assistance of Dr.

Dennis Clifford, Environmental Research Department, University of

Houston, Houston, Texas. This process reduces the biochemical oxygen

demand (BOD) of the wastewater. In general, the organic wastes that

produce BOD are decomposed by bacteria, in the presence of nutrients

and dissolved oxygen, into carbon dioxide and water, with the production of more bacteria. Marine algae grow in this nutrient-rich

medium. The bacteria that die and the algae also contribute to the

BOD. The liquid fraction of this decomposition and some suspended

solids are discharged after treatment with hydrogen peroxide (Fig.

12). The solids that precipitate are retained as activated sludge to

re-seed the process.

An array of polyvinyl chloride (PVC) pipes 3.8 cm (1.5 in) drilled

at 2.54 cm (1 in) intervals with 0.4 cm (0.016 in) holes was placed

on the bottom of each digestion tank and used to keep the wastewater

9
vigorously aerated. Air is supplied to the pipes by two Cyclonaire

blowers (Table 3). After aeration of the wastewater for 20 hr, the

blowers are turned off and the suspended solids allowed to settle for

4-6 hr. Thereafter, the supernatant effluent is discharged through a

10.2 cm (4 in) PVC pipe at a level of 25.4 cm (10 in) above the bottom

of the tank. The discharge pipe is connected at least 25.4 cm (10 in)

above the bottom of the tank so that the layer of activated sludge

near the bottom is not drawn off at the time of discharge. When all

15 raceways are in use, the wastewater effluent is discharged three

times a week into a storm sewer at a rate of about 47,100 liter

(12,450 gal) per discharge.

Hydrogen peroxide (H202) is injected as the wastewater effluent is

discharged. Concentration of the injected H202 is 60-100 ppm in the

effluent discharged through the 10.2 cm pipe. The H202 is applied

with an injector pump (Fig. 12) outfitted with a calibrated flow meter

(Table 3).

A 2.54 cm (1 in) sampling pipe was installed in the 10.2 cm

wastewater drain line approximately 12.2 m (40 ft) from the digestion

tanks (Fig. 13). During each discharge cycle, a three-aliquot sample

of the treated wastewater effluent is taken to determine average salinity, pH, total organic carbon (TOC), and total suspended solids (TSS)

(Table 4). Aliquots of 250 ml each are taken at the beginning, midpoint and end of the discharge cycle. The samples are analyzed for

TOC by a private contractor. The pH is determined with a pH meter.

TSS are determined for a 250 ml aliquot of effluent using GF/C filters

and a millipore filter. The filtrate is dried in an oven at 1050C and

weighed. The results of TSS and TOC analyses (Table 3) are reported

monthly (in pounds/day) to the Texas Department of Water Resources,

Water Quality Board, Austin, Texas under permit no. 02299.

After the wastewater effluent discharge is completed, the

discharge lines are flushed with 1,136 liter (300 gal) of freshwater

to which had been added 1.5 liter of H202. Currently, the treated

wastewater effluent is discharged into a municipal storm sewer. Each

10
digestion tank also is plumbed from the bottom into the City of

Galveston domestic sewerage system (Fig. 13). At the end of each

annual head start period, the residual sludge is washed from the

digestion tanks into this system.

HEAD START OPERATIONS

Receipt of Hatchlings - At the Padre Island National Seashore, hatchlings are packed in wax-coated, cardboard boxes (58.4 cm or 23 in

long, 40.6 cm or 16 in wide, and 25.4 cm or 10 in deep) containing

polyurethane foam padding soaked with water to prevent dessication of

the hatchlings. They arrive at Galveston after 2-6 hr in shipment.

NPS personnel pack the boxes in such a way that each clutch is kept

segregated from others. Upon arrival at the head start facilities,

the hatchlings are first rinsed with seawater and then inspected

closely for abnormalities and mortality. The most common abnormalities observed are: "cross-beak" (upper and lower jaws grossly

malformed, left eye missing); concave plastron (plastron grossly

depressed); curvature of the spine; shortened spine (turtle much

greater in width than in length); plastron improperly healed (unclosed

yolk-sac attachment site) and deformed flippers. Turtles with abnormalities are isolated in the sick bay. Turtles with improperly healed

plastrons are treated with a topical antibacterial ointment

(Terramycin, Gentamycin, Neomycin, or Furacin), and in most cases this

eventually heals. Abnormal turtles that have survived usually have

been transferred to Texas A&M University, College Station, Texas, or

to the University of Texas Institute of Marine Science, Port Aransas,

Texas, to be used for research. A small number of these have been

transferred by Texas A&M University to Sea Turtles, Inc., directed by

Mrs. Ila Loetscher, South Padre Island, Texas.

11
weighing and Measuring - Either before they are transported to

Galveston or immediately upon their arrival (i.e., during the rinsing

and inspection processes), all hatchlings are weighed (wet weight) and

measured (carapace length and width; as recommended by Bjorndal and

Balazs 1983). Similar measurements are made on random samples taken

once per month thereafter and are recorded to the nearest 0.1 g. It

has been determined that a minimum of 25 turtles, by clutch, is an

adequate sample of turtles from a given raceway per month (Caillouet

et al.~). During the weighings, no attempt is made to dry the

turtles. After consecutive weighings of three turtles, the balance

pan is dried and re-zeroed. An Ohaus, triple-beam balance is used.

Assignment of individual turtles from a given clutch to a given

raceway and distribution of portions of clutches to different raceways

currently is determined according to experimental design. In the

past, clutches were placed in raceways more or less sequential.ly as

they were received from NPS, starting with raceway 1 and progressing

consecutively among the numbered raceways.

Clutch identity of each turtle is monitored through the bucket

identification code. The numbering system used within a raceway is

shown in Fig. 14. For instance, the bucket marked with an X in Fig.

14 is labeled 83-1-G-4. This code indicates that the turtle was from

the 1983 year-class and was located in raceway 1, in the bucket in row

G and column 4. Such codes, once assigned to individual turtles, are

used throughout the head starting process to track individual turtles.

In this way, the history of a given turtle can be tracked back through

head starting for information on its maternal parentage and details

about egg laying, egg collection, incubation, hatching, growth, amount

of food fed, health care, etc. For tagged turtles released, the tag

number can be linked with the identification code used during head

starting. Computerized data files carry this identification code to

track data for each turtle.

12
Feeding - The food used in the head start project has been a commercially prepared, dry, pelleted diet (Table 5) manufactured by central

Soya and Subsidiaries, Fort Wayne, Indiana. In 1978, the first year

of operation, natural foods such as lettuce, fillet of croaker, fillet

of freshwater sawfish and fillet of whiting were fed (McVey et al.~),

but the cost and inconvenience of using such foods are prohibitive.

The diet prepared by Central Soya contained 45% crude protein, 5%

crude fat, 3% crude fiber, and 47% other ingredients (including, but

not necessarily limited to, moisture). This provided good growth

(Fig. 1) and survival (Table 1) of the turtles.

One problem encountered with the Central Soya food was its contamination by small, unidentified beetles when it was stored in a dry,

air-conditioned room. Apparently, the food itself was contaminated

with the eggs of the insect, but this was never confirmed. The

problem with the beetles was eliminated, beginning in 1982, by keeping

,the food in frozen storage until use. Floating pellets are essential,

both to facilitate feeding at the surface and to avoid loss of food

through the holes in the bottom of the buckets. The hatchlings and

older turtles are capable of diving to the bottom of the buckets to

feed, but the feed disintegrates in time. Thus, surface feeding is

encouraged, and this requires floating food.

Feeding of hatchlings must be carefully monitored as overfeeding

can result in compaction of the gut that may lead to death. Because

ilMcvey, J. P., J. K. Leong, R. S. Wheeler and R. M. Harris. The

culture of young Kemp's ridley sea turtles (Lepidochelys kempi).

Unpublished manuscript, NMFS SEFC Galveston Laboratory, Galveston,

Texas.

13
sacrificing live, healthy hatchlings is prohibited, no direct way has

yet been developed to determine when the yolk material has been adequately absorbed so as to determine when feeding should start. During

the first five years of head starting, commencement of feeding was

determined subjectively based on the apparent willingness of hatchlings to accept food, usually at 5-7 days of age.

Twelve apparently healthy hatchlings of the 1983 year-class were

isolated in basins in the sick bay, and were weighed on a Mettler

balance every day (at the same time of day) for 18 days. Over a

period of 17 days, none of the twelve hatchlings lost weight; in fact,

their average weight went from 17.3 to 21.4 g. The study was terminated on the 18th day because of concern for the overall health of

the hatchlings. One of the test turtles died two days after the study

was terminated. Necropsy showed that this turtle died from an invasive fungal infection, rather than from lack of food. Interestingly

enough, there appeared to be no significant reduction in the overall

volume of its yolk-sac.

The rate at which head started turtles are fed is based on the

average weight of a sample of surviving turtles at monthly intervals

during head starting. Average weight was estimated by weighing a randomly selected sample of 375-450 turtles (25-30 per raceway). The

initial rate of feeding for hatchlings is roughly 5% of body weight.

This rate is subjectively changed each month until a rate of roughly

1.5% of body weight is reached for yearlings. The daily food ration

is usually divided into two equal portions, one fed in the early

morning and the other in late afternoon. The total amount of food to

be fed to the turtles on a given day is calculated by multiplying the

average weight of the sample of turtles by the total number of survivors. The rations of food are distributed by personnel to the

turtles by hand, subjectively attempting to take into account the size

of each turtle, by feeding more or less food to the larger or smaller

turtles, respectively.

14
Growth - Growth and survival of head started Kemp's ridleys of the

1978-1983 year-classes has been described in detail by Caillouet et

al.2/. We have plotted the arithmetic average weight (kg) against

elapsed time (days) for the 1978-1983 year-classes in Figure 1.

Health Care - During head starting, each turtle receives a precursory

examination for evidence of disease during the twice daily feeding.

Once every ten days, each turtle is examined closely for signs of

disease and/or injury. Any turtle displaying signs of disease or

injury is isolated at that time in the sick bay. Those with minor

ailments, such as skin lesions, are treated in the sick bay; others

with more serious problems are submitted to the pathologist, Dr. Jorge

Leon~/, for clinical diagnosis and treatment (Clary and Leong 1984).

Many of the prophylactic, diagnostic and therapeutic methods used

during head starting of Kemp's ridleys were developed or adapted by

Dr. Leong and his staff. Signs of turtle diseases and actions taken

were as follows:

a) Apparent disorientation - The condition is noted and observations

are made on the turtle twice daily. If the condition persists for

more than 48 hours the pathologist is consulted.

b) Superficial dermal and shell lesions - Most lesions respond to

daily topical applications of Terramycin (oxytetracycline hydrochloride ointment combined with polymyxin B sulfate). More persistent lesions were treated with Furacin (nitrofurazone), a

topical antibacterial agent, but we use it as a last resort

because the polyethylene glycols present in the base can be

~Leong, J. K., D. L. Smith, D. B. Revera and J. C. Clary III.

Health care and diseases of captive hatchlings of the loggerhead

and Kemp's ridley sea turtles. unpublished manuscript, NMFS SEFC

Galveston Laboratory, Galveston, Texas.

15
absorbed through the skin and may cause renal impairment. If

lesions do not appear to respond to treatment within 5 days, the

pathologist is consulted.

c) Buccal or anal hemorrhaging - The pathologist is notified at once

and the entire raceway is isolated from the general seawater

supply and wastewater discharge systems. This condition is indicative of a potential epizootic of bacterial etiology with

possible severe consequences (e.g., hemorrhagic septicemia leading

to death). The entire head start staff is placed on alert and

feeding of turtles in the affected raceway(s) is discontinued.

d) Cloaccal prolapse (tissue extruding from the anus of the turtle) -

Antibacterial ointment is applied topically to the affected area

for 48 hours; if there is no improvement, the pathologist is

notified.

Tagging - The tags used for all head started sea turtles are Hasco

type, style 681, self-piercing, self-clinching, ear tags, manufactured

from monel metal (National Band and Tag Co.). They are inscribed with

a sequential letter-number code as well as the message "Send NMFS Lab,

Virginia Key, Miami, FL 33149." Tagging with this tag is usually done

about 30 days prior to release of the turtles to allow remedial action

in case of'tag loss, infection, or ~ny tag-related mortality. The

tags are normally inserted on the trailing edge of the right front

flipper; double-tagged individuals also are tagged on the left front

flipper (Fig. 15). The tags used on the 1978-1981 year-classes were

first soaked in gasoline for 24 hours to remove any oil or grease,

then in 90 percent ethanol for 24 hours prior to use. Tags used with

the 1982 year-class received no treatment prior to application, and an

outbreak of hemorraghic septicemia ensued. Thereafter, the tags used

on the 1983 year-class were sterilized by autoclaving prior to

tagging.

16
The area of tag insertion on the flipper is swabbed with tincture

of iodine prior to tagging, then Neosporin, a broad-spectrum antibacterial ointment, is applied to the tip of the sharp clasping device of

the tag before the tag is inserted. A heavy, cast-iron, tagging tool

supplied by National Band and Tag Co. is used to affix the tag to the

flipper. It is sometimes necessary to recrimp the tag with pliers to

secure it. Careful observations of tag numbers, body weight, carapace

length, carapace width, and gross observations of general turtle condition and health are made and recorded as each tag is applied. This

usually has been the last time that measurements and weights of the

head started turtles were recorded before the tagged turtles were

released. Turtles do not actively feed for 1-2 days after tagging, so

feeding is discontinued for 48 hours. This procedure eliminates

fouling the water with uneaten food. Some head started turtles of the

1978, 1979, and 1980 year-classes were outfitted with radio transmitters. The results of tracking studies on these turtles have been

reported by Klima and McVey (1981), McVey and Wibbels (1984), and

Wibbels (1984).

Transportation to Release sites - Head started Kemp's ridleys are

transported to release sites or marine aquaria in wax-coated, cardboard boxes (the type used to transport hatchlings from Padre Island

to Galveston). The boxes are modified by partitioning them with

plywood to make two horizontal "layers" within each box (Fig. 16).

Two 1.27 em (1/2 in) air-holes are drilled at each end of the box and

the floor of each layer is covered with a piece of 1.27 em thick

polyurethane foam to cushion the turtles. The foam is moistened to

prevent dessication of the turtles during transit. Eight yearling

turtles are transported in each box, four turtles to a layer. Turtles

are oriented with their heads toward the corners to prevent them from

biting each other, and to place their heads near the air-holes. The

lid of the box is secured with gray, duct tape, one piece completely

around the length of the box and another completely around the width.

17
Copies of Federal and state permits are attached to each box.

Normally, a crew of ten persons requires 3 hours to pack and load

1,600 tagged yearling turtles for shipment. Packing and loading

should be done as quickly as possible to reduce the amount of time

that turtles are held in the boxes.

Release - The release sites, dates of release, and tag series used for

release of head started Kemp·s ridley sea turtles from the 1978-1983

year-classes are presented in Table 6. Most tagged turtles of the

1978 and 1979 year-classes were released in Florida waters because

most yearling Kemp·s ridleys had been observed to occur in that area

(Klima and McVey 1981; McVey and Wibbels 1984). Some of the 1978 and

1979 year-classes were released off Padre Island (ibid.). Most of the

1980-1983 year-classes were released offshore of Padre and Mustang

Islands, Texas, in hopes of reinforcing any imprinting they might have

received as eggs and hatchling~. Some of the 1980 year-class were

released off Campeche, Mexico, and some of the 1982 year-class were

released in Nueces Bay and off Sabine Pass, Texas. Most of the 1980

and 1981 year-classes were released from the TPWD research vessel

"WESTERN GULF". Most of the 1982 year-class and all of the 1983 yearclasses were released from the U.S. Coast Guard cutter "POINT BAKER".

The portion of the 1982 year-class released in Nueces Bay, Texas, was

released from a TPWD boat. Three groups of turtles tagged with radio

transmitters were released from small boats and were tracked by

aircraft (Klima and McVey 1981; McVey and Wibbels 1984; Wibbels 1984).

Tagged turtles are usually released by dropping them over the

side of the vessel into the water. Almost without exception, they

will beat their flippers so rapidly that they give the impression of

"flying" across the surface of the water, sometimes to distances

estima~ed to be around 30.5 m (100 ft), before submerging.

18
SEA TURTLE HEAD START RESEARCH AND GENERAL OBSERVATIONS

Recaptures of Head Started/Tagged/Released Turtles

In general, tagged Kemp's ridleys have become widely dispersed in

U.S. waters, from Brownsville, Texas, along the coast of the Gulf of

Mexico, to 'the U.S. eastern seaboard as far north as Long Island, New

York (Klima and McVey 1981; Caillouet 1984; McVey and Wibbels 1984;

Wibbels 1984; Fontaine et al.~/). Foreign recoveries have been

reported from France, Morocco, Dominican Republic (unconfirmed), and

the Mexican coast of the Gulf of Mexic~/. Although the reports of

measurements on recovered turtles are scanty and not always reliable,

available data indicate good growth rates (averaging 5 g per day) of

the head started, tagged Kemp's ridleys in their natural

environmen~.The turtle at large for the longest period (1,563 days

or 4.3 yr) was from the 1978 year-class, and was recovered from

Hunting Island, South Carolina, on August 18, 1983 after release from

Homosassa, Florida on May 8, 1979.

Most tagged Kemp's ridleys have been recovered alive (Table 7) and

returned to the environmen~/; however, in many cases the flipper tag

was removed before the turtle was re-released because the inscription

on the tag encourages such removal. Consideration should be given to

changing the message inscribed on the tags so that the recovery information is retained, but the tag is left intact when a live turtle is

re-released.

Of those turtles from the 1978 and 1979 year-classes released in

Florida, all recoveries from the former year-class and most of the

recoveries from the latter year-class have been made on the east coast

of the United States as far north as Long Island, New Yor~. Carr

(1980) postulated that juvenile Kemp's ridley sea turtles found in the

Atlantic Ocean, particularly those along the Atlantic coasts of North

America and Europe, are lost "waifs" that never return to the Gulf of

Mexico. Head started Kemp's ridleys survive, overwinter, and grow

normally in the Atlantic, and they may return at some later date to

19
the warmer waters of the Gulf of Mexico where the breeding population

of Kemp's ridleys is foun~. Most recoveries from the 1980-1983

year-class released off Padre Island have been made in the western

Gulf of Mexico.

Feeding Regimen and Food Utilization - After 1980, all year-classes

were fed only a dry, pelleted diet (Table 5). The utilization of

this foodstuff, incidence of feeding, and rate of feeding have been

investigated for the 1978-1981 year-class by McVey et al.~. In

general, our observations indicated that the gross food conversion

ratio (total weight of food fed:total amount of weight gained) for

turtles fed this commercially prepared diet was good, ranging from

1.3:1.0 to 1.7:1.0 in the 1982 and 1983 year-classes, respectively.

This is particularly impressive since all the food offered was not

always eaten. The Kemp's ridley sea turtle evidently possesses very

efficient digestive and metabolic systems. It would appear from these

data that Kemp's ridleys head started in buckets at the Galveston

Laboratory require a very small amount of food to meet daily dietary

needs for maintenance. Confinement in buckets may limit the scope of

activity and thereby reduce the expenditure of energy (Caillouet et

al.~/). Turtles head started in captivity gained at an average rate

of only 3 g per day during the first year of life. This was slower

growth as compared to post-release growth in the natural environment

(5 g/day), but it is to be expected if the animals released were still

in the exponential phase of growth for a time after their release. In

general, growth in captivity is more rapid than growth in the wild

(Caillouet 1984; McVey and Wibbels 1984; Caillouet et al..!/).

Food Color Preference - During the manufacture of the fiberglass

basins, one basin evidently was splattered with a drop of bright

red paint. We noticed that any turtle placed in this basin would

invariably "worry" or peck at this spot continually during daylight

hours. Further, we observed that hatchling and yearling turtles

20
reacted (by rapid movement) to personnel dressed in red, orange,

yellow, or white, while they responded much less to those dressed in

darker colors such as black, green, blue, or brown.

Therefore, we designed a study to test possible preferences of

head started Kemp's ridleys for foodstuff colored with different food

dyes (Appendix 1). pieces of fresh, abdominal muscle of shrimp were

soaked overnight in Kroger, Inc. food coloring (either red, yellow,

blue, or green). pieces of colored food and non-colored (control)

food were affixed to a special food holder (Fig. 17) in randomized

order. Four colored (red, yellow, blue and green) pieces and one

uncolored (control) piece of shrimp were presented on the holder to

each of 20 turtles in each of three different consecutively tested

groups. The pieces of shrimp were placed on the holder randomly for

each turtle tested, so that their location on the holder would not be

confounded with treatments (different colors and control). Each

turtle was allowed to bite at the food twice to determine its first

and second choices. After the first choice (or bite) was made, the

holder was flipped over so that the order of food on the holder was

reversed before the second choice or bite was made. Any food item

removed on the first bite by the turtle was replaced at the same position by an item of the same color as that removed.

The dominant choice of colored shrimp muscle was for pieces

colored red, for both first and second choices (Table 8). possibly

the turtles were also reacting to a chemical stimulus created by the

food coloring dye itself, as the chemical makeup of the different

colors of food dyes is not the same. However, we believe that the

choice was based primarily on optical rather than olfactory or other

chemo-sensory mechanisms. This is reinforced by the previously

observed reactions of turtles to the red spot in one of the basins,

and to colored clothing worn by project personnel.

21
celestial orientation of Hatchlings - Another study was conducted in

cooperation with the Burke Baker Planetarium in Houston, Texas, to

test celestial orientation in head started hatchling Kemp's ridley sea

turtles of the 1983 year-class. The primary objective of the study

was to determine if hatchlings that had never seen the stars would

demonstrate any ability to orient relative to celestial patterns

(assuming they had not seen stars before we received them; they were

kept inside the boxes during transit, and in the quonset hut thereafter) •

Observations were made on 4 different hatchlings, each tested

alone. A hatchling was placed in a submersible pen positioned in the

center of a fiberglass basin (similar to the ones in the standing

basin unit) located within the planetarium and oriented on a compass

rose for true magnetic north. After a turtle had acclimated for 15

minutes within the small pen, the pen was released and sank to the

bottom "freeing" the turtle. After the turtle was released, two types

of observations were made: (1) the initial position to which the

turtle traveled on the periphery of the basin; and (2) the position

of the turtle around the periphery of the basin at 1 min intervals.

The number of observations depended upon the amount of time allotted

for use of the planetarium. All four turtles used in this study were

tested on four different occasions: July 28, 1983, August 4, 1983,

August 11, 1983, and September 8, 1983, and two of them were tested

twice on an additional day, September 16, 1983.

The initial position to which the turtles traveled (Table 9)

appeared to be random with regard to east-west orientation (10 to the

west and 9 to the east), but the direction traveled after initial contact with the side of the test basin was predominantly in a clockwise

motion; however, after the first few minutes all turtles would randomly "wander" around the periphery of the test basin. The observations of orientation of turtles within the basins with time (Table

9) showed that the test turtles appeared in the western half of the

test basin more often than in the eastern half (62%:38%,

22
respectively).

We recognize that these observations are insufficient to draw

definite conclusions. It would appear from these preliminary data

that head started turtles showed a particular preference for the

western half of the test basin as compared with the eastern half.

Whether or 'not this represents celestial orientation remains to be

determined.

Re-Cycled Seawater - The seawater system at the Galveston Laboratory

does not have the pumping or storage capacity to allow for flowthrough at rates sufficient for daily replacement of water in all

of the raceways (approximately 520 gal/hri 33 ml/min). The water

quality deteriorates rapidly prior to cleaning the raceways (Fig. 18),

due to accumulation and decomposition of turtle waste products and

uneaten food. Therefore, we investigated the feasibility of using recycled seawater. The system tested included an external Permute tube

settler and a rotating biological contactor (biodisc). In general,

this system significantly reduced the rate of increase in concentration of dissolved organics, particularly that for total ammonia, as

measured by ammonia-nitrogen (Fig. 19). While this system seems promising, flow-through systems might be preferable for head starting sea

turtles, but have the possible disadvantage of inflating costs of

heating the seawater.

The laboratory procedures used for analyses of total ammonia,

nitrite, nitrate, and phosphate in seawater are presented in Appendix

2. Descriptions of various seawater treatment systems and biological

operation of such systems are presented by Wheaton (1977).

Tagging - Various tagging methods have been investigated during head

starting of the Kemp's ridley sea turtles. Double-tagging with

monel metal flipper tags was first tried on 54 turtles of the 1981

year-class and later on 400 turtles from the 1982 year-class to

increase the probability that at least one flipper tag would be

23
retained. As of October 1, 1984, 32 individuals (7.5%) of 424

doubled-tagged turtles released off the Texas coast had been

recovered, as compared with 191 (7.9%) recovered out of 2,428 singletagged animals released from the 1980 and 1982 year-classes combined.

However, the double-tagged turtles that were released into the

environment have not been at-large long enough to ascertain the value

of the second tag.

Another observation was that double-tagged turtles were more

lethargic after tagging and took longer to begin feeding after tagging

than those that received a single tag. Double-tagging appears to be

more traumatic. The tags used for tagging the 1982 year-class were

not sterilized and an infection ensued. This problem probably can be

avoided by sterilizing the tags before tagging.

A "living-tag" technique developed by Dr. John and Mrs. Lupe

Hendrickson, University of Arizona, Tucson, Arizona, involves surgical

removal of pieces of plastron and carapace, interchanging the grafts,

and securing them with histological glue. As the turtles grow, the

lighter colored plastron transplant makes a vivid mark on the darker

background of the carapace.

Three "living-tag" experiments were conducted. The first two were

conducted in cooperation with Dr. and Mrs. Hendrickson and the last by

NMFS personnel. The first was done on 200 turtles ("living-tagged" on

right coastal skutes 2, 3, or 4) of the 1980 year-class and the second

on 432 turtles ("living-tagged" on left coastal skute 3) of the 1982

year-class. In the third experiment, all turtles from the 1983 yearclass were "living-tagged" on left coastal scute four. The lightercolored plastron transplant contrasted with the darker carapace within

the first 90-120 days after tagging turtles 5-7 months old (first and

second experiments). After this time, the mark, in some cases, began

to fade or blend in with the surrounding carapace that becomes lighter

in color as the turtle ages. Retention of the mark was not good in

the second experiment (1982 year-class; Table 10).

Twenty-five head started Kemp's ridleys of the 1982 year-class

24
were sent to five public marine aquariums, five per location (Table

2). Each turtle had been tagged with an internal, binary-coded, metal

tag manufactured by Northwest Marine Technology Co., Shaw Island,

washington. The tag was inserted beneath the skin of the right front

flipper near the distal end of the humerus (Fig. 20). Since these

turtles are being held in captivity beyond their first year, observations can be made on tag retention, tag migration (if any) within

the tissue, and tag detection and decoding with x-ray equipment. Some

of these turtles also are marked with the "living tag" allowing opportunity for examination as they grow older and carapace color lightens.

General Observations - Kemp's ridleys having dermal lesions were

placed outside in sunlight for 2-4 hr. This exposure to sunlight

seemed very effective for treating certain skin lesions. Further,

turtles that became sick during head starting and then received the

"sunshine treatment" appeared to be much more active, had better appetite, and were, in general, in better overall condition than sick

turtles that did not receive the treatment.

One problem encountered was excess growth of filamentous green

algae on the young turtles. In early years, the turtles were periodically scrubbed with soft brushes to remove this algae. Starting in

1981, this practice was discontinued because it was thought that

scrubbing might remove natural protection, such as a mucus-layer. The

algae growth probably was stimulated by the excessive levels of nitrogenous compounds within the holding water and by ample sunlight (90%

of the incident sunlight) that penetrated the polyethylene covering of

the quonset hut. At 7 to 8 months of age, or at a body weight near

454 g (1 lb), the turtles began to lose much of the attached algae.

with turtles held for longer periods we noticed that by the age of 12

months all normal sized turtles had lost the attached algae. It may

be that, beyond a certain size or age, the Kemp's ridley's produce

epidermal substances that prevent fouling growth on the shell. This

is suggested as an area for further research.

25
One of the more unusual and unanticipated behaviors was the

variation in the reaction of head started turtles to different personnel working within the project. Most of the time, the turtles, especially yearling and older, would react violently when picked up. In

most cases, when picked up by hand, the turtles would rapidly beat

their flippers and attempt to bite the person holding them. Also,

when the turtles were larger and held in standing basins, they would

splash water on a person approaching their basins. The turtles did

not react in this way to one of the coauthors (Kathy L. Williams

Indelicato) of this report. When she picked up any of the turtles

they would immediately become very still and would not move until

placed back into their buckets. Also, regardless of the type or color

of clothing she wore, the larger turtles in the standing basins would

not react and would float very still on the surface until Mrs.

Indelicato had passed. This observation and others are anecdotal,

but, for those involved in caring for and handling the aggressive

Kemp's ridley sea turtle, they may be of interest.

DISCUSSION

Survival and growth of head started Kemp's ridleys in captivity

during the first six years of the project are shown in Table 1 and

Figure 1 (see also Caillouet et al.1/). The combined survival rate

for the six year-classes was 82%. The survival and growth of head

started Kemp's ridleys in the wild appears to be good (Fontaine et

al.1V), but it is too early to gauge the success of maturation, copulation and nesting of such animals in the wild (Caillouet 1984).

Recently, egg laying was reported in two 5-year-old, head started

Kemp's ridley maintained in captivity at Cayman Turtle Farm [1983]

Ltd. (Wood and Wood, 1984). According to Wood and Wood (1984), more

than 60 eggs were laid by the sea turtles in May 1984. Three of the

eggs in one clutch hatched, indicating that copulation and fertilization had occurred. Unfortunately, the three hatchlings did not

26
live. Nevertheless, this represents a breakthrough in mating and

nesting of captive individuals, indicating that head started animals

might contribute to a captive breeding population as a "safety net"

for the species (Caillouet, 1984). The potential for maturation and

copulation among captive breeding populations of Kemp's ridleys is now

less remote than was once thought. The survival and fast growth of

eight out of ten head started ridleys from the 1978 year-class at

Sea-Arama Marineworld in Galveston (Fig. 21) is very encouraging

(McVey and Wibbels, 1984; Caillouet et al.~/).

After six years of involvement in head starting Kemp's ridley sea

turtles, we believe that there are four areas of research that should

be emphasized in future work:

(1) Reproductive physiology and behavior as related to reproduction in the wild and in captivity.

(2) Tags and tagging of sea turtles. None of the tags presently

being used adequately meet the needs. A tag or mark is

needed that will last the entire life of the tagged turtle,

and that can be easily recognized and identified by whomever

recaptures a Kemp's ridley. Further, through publicity and

through changes in the message on the tag, the finder of a

tagged turtle must be encouraged to take the needed observations and report them in a timely manner without removing

the tag from live animals. Removal of the tag from dead animals should be encouraged as part of the reporting of

stranded sea turtles.

(3) Sex determination. A simple technique must be developed to

determine the sex of live hatchlings and older immature individuals without injuring them. Sex determination is essential for proper management and conservation of this

endangered species, because sex ratios of hatchlings may be

27
affected by incubation temperature. The hormonal studies by

Dr. David Owens, Texas A&M University, and his students are

encouraging in this regard. Laparoscopy also has been used

successfully by Owens and his students on larger juvenile and

adult turtles, but neither technique is simple nor applicable

to hatchlings.

(4) prevention, recognition, diagnosis, and treatment of diseases

among captive populations of Kemp's ridleys must be improved.

The successful establishment of captive breeding populations

of Kemp's ridleys will, in our opinion, depend to a large

extent upon solutions to the disease problems encountered in

the husbandry of this species. Improvements in seawater

systems toward providing high quality seawater for captive

rearing of sea turtles would go far toward reducing disease

problems. Other environmental influences (e.g., sunlight,

temperature, etc.) also may affect the incidence of disease.

Good nutrition is another important factor.

EPILOGUE

Prior to arrival of the first group of Kemp's ridley sea turtle

hatchlings at Galveston in 1978, very little was known about the husbandry of this endangered species (r4rquez 1972; Pritchard and Marquez

1973). Little if anything was known of their spatial needs, what to

feed them, how to recognize or to treat their diseases, nature of

their behavioral characteristics, seawater quality requirements, or

any of their other day-to-day needs. Information was available from

comparable work done on other sea turtle species, but it was soon

apparent that many of these procedures were not applicable to the

Kemp's ridley. Consequently, many of the techniques and equipment

developed, adapted and used at the NMFS Galveston Laboratory were the

results of experimental and trial-and-error approaches as well as

28
serendipidous findings. For instance, in 1978 no one expected the

aggressive behavior, exhibited among Kemp's ridley hatchlings, that

necessitated isolation of each hatchling in an individual container

(Klima and McVey 1981). The suspended, plastic bucket system that

evolved seems simple in hindsight. However, its development required

considerable observation and effort on the part of Bonnie Cockrell,

Clark Fontaine, Larry Lansford, Jorge Leong, Cornelius Mock, Dickie

Revera, and Ray Wheeler over a period of time (Clary and Leong

1984!1) •

The head start facilities themselves have been modified a number

of times during the tenure of the project (Fig. 22). The changes in

1981 were made to provide a sick bay, to improve wastewater treatment,

to provide a maintenance shop and tool storage areas, and to increase

seawater storage capacity. The modifications following Hurricane

Alicia in 1983 were made in an effort to reduce utility costs through

more efficient heating, to facilitate replacement of polyethylene

covers, and in an attempt to provide better seawater quality.

All authors of this report, at one time or another, were closely

involved in the day-to-day care and husbandry of Kemp's ridleys, and

they feel honored to have been a part of the international effort to

save this endangered species from extinction. Our sincere hope and

wish are that the head starting of Kemp's ridleys be continued. We

support and encourage all efforts toward the conservation of the

Kemp's ridley sea turtle. William Beebe made the apocalyptic observation that "when the last animal of a race of living things breathes

no more, another heaven and earth must come to pass before such a one

can be again."

ACKNOWLEDGMENTS

Work on the 1978-1983 year-classes of Kemp's ridleys was conducted

under consecutive INP permits no. ABC-IV-0751/8676, ABC-IV-1258/4287,

242-0180-3794, 242.2-2610, 242.2-00026/1147 and CG-115-900 (and diplo29
matic note no. 316502), respectively, under FWS Endangered and

Threatened Species Permits no. PRT 2-1770, PRT 2-4481 and PRT-676379,

and under scientific permits issued by TPWD and FDNR.

Special thanks are due Ren~ ~rquez Mill~n (INP, Mexico), Jack

woody and David Bowman (FWS, Albuquerque, New Mexico), Milford

Fletcher (NPS, Santa Fe, New Mexico), Patrick Burchfield (Gladys

porter Zoo, Brownsville, Texas), and Robert King (NPS, Corpus Christi,

Texas), their staffs and their predecessors, for their respective

roles in providing Kemp's ridley hatchlings for head starting.

Dr. James P. McVey, Mr. Ray S. Wheeler, and Mr. Larry Lansford

worked diligently to establish the sea turtle head start research

facilities and project from 1978 to 1981. Bonnie Cockrell, Thane

Wibbels, Jean MCDonald, Diana Nunez, Jody Gruber, Joey Cheeks, and

Cornelius Mock contributed much to establishment and improvement of

systems, methods and equipment. Dr. Jorge K. Leong was instrumental

in developing the facilities for and in recommending treatments for

diseases of head started turtles, as well as providing health care for

the 1978-1981 year-classes, along with his staff including John Clary

III and Dickie Revera. The administrative guidance and support provided by Drs. Charles W. Caillouet, Jr. and Edward F. Klima have been

both stimulating and encouraging.

A special thanks goes to Mrs. Carole H. Allen and all the many

supporters of HEART (Help Endangered Animals - Ridley Turtles), a

special committee of the Piney Woods wildlife Society, North Harris

County College, Houston, Texas, whose diligent and conscientious

efforts have enhanced public awareness and support for the conservation of Kemp's ridley sea turtles.

We thank John Kerivan and his staff at Sea-Arama Marineworld in

Galveston, Texas, for maintaining 10 head started Kemp's ridleys at

their facilities and for providing data for our paper.

Dickie Revera and Sharon Manzella assisted in checking, verifying

and validating some of the data used in our paper.

Dr. John and Lupe Hendrickson, University of Arizona, Tucson,

30
Arizona, demonstrated the "living-tag" procedure and assisted in

experiments involving "living-tags".

The figures and illustrations for this manual were prepared by

Daniel Patlan and the manuscript was typed by Bea Richardson.

31
Disease studies aid Kemp's

Herpetological Review

LITERATURE CITED

Bjorndal, K. A. and G. H. Balazs (editors). 1983. Manual of sea

turtle research and conservation techniques, Second Edition.

Center for Environmental Education, Washington, D.C., 126 pp.

Caillouet, C. W., Jr. 1984. Essai de pr~vention de l'extinction

de la tortue de Kemp. Les Carnets de Zoologie (Bulletin of the

zoological Society of Quebec) 44(2):28-34.

Carr, Archie. 1980. Some problems of sea turtle ecology. Am. Zool.

20:489-498.

C~vez, Humberto, Martin Contreras G., and T. P. Eduardo Hernandez D.

1968. On the coast of Tamaulipas. International Turtle and

Tortoise Society Journal Part one, 4:20-29, 37; Part two, 5:16-19,

27-34.

Clary, J. C. III and J. K. Leong. 1984.

ridley sea turtle head start research.

15 (3):69-70.

Fontaine, Clark, Jorge Leong, and Charles Caillouet. 1983. Head

starting Kemp's ridleys, 1982, p. 22-29. In: David Owens, Diana

Crowell, Gayle Dienberg, Mark Grassman, Sheilah McCain, Yuki

Morris, Nancy Schwantes, and Thane Wibbels (editors), Western Gulf

of Mexico Sea Turtle Workshop proceedings, Texas A&M University,

Sea Grant College Program, College Station, Texas, TAMU-SG-84-105,

74 p.

Hildebrand, Henry H. 1963. Hallazgo del ~rea de anidaci~n de la

tortuga marina "lora", Lepidochelys kempi (Garman), en la costa

occidental del Golfo de M~xico. Ciencia (~xico) 22(4):105-112.

32
Klima, Edward F. and James P. McVey. 1981. Head starting the Kemp's

ridley turtle, Lepidochelys kempi. In: Bjorndal, Karen A.

(editor), Biology and Conservation of Sea Turtles, Proceedings of

the World Conference on Sea Turtle Conservation, Smithsonian Inst.

Press, Wash., D.C., 583 p.

Karquez, Re~. 1972. Resultados preliminares sobre e dad y

crecimineto de la tortuga lora. Lepidochelys kempi (Gorman).

Men. IV Congreso Nac. Ocean. (Mexico): 419-427.

M'arquez, Ren~. 1983. Current status of the Kemp's ridley population.

In: David Owens, Diana Crowell, Gayle Dienberg, Mark Grassman,

Sheilah McCain, Yuki Morris, Nancy Schwantes, and Thane Wibbels

(editors), Western Gulf of Mexico Sea Turtle Workshop proceedings,

Texas A&M University, Sea Grant College Program, College Station,

Texas, TAMU-SG-84-105, 74 p.

I

Marquez, R., A. Villanueva o. and M. Sanchez P. 1981. The population

of the Kemp's ridley sea turtle, Lepidochelys kempi. In:

Bjorndal, Karen A. (editor), Biology and Conservation of Sea

Turtles, proceedings of the World Conference on Sea Turtle

Conservation, Smithsonian Inst. Press, Wash. D.C., 583 p.

McVey, J. P. and T. Wibbels. 1984. The growth and movements of

captive-reared Kemp's ridley sea turtles, Lepidochelys kempi,

following their release in the Gulf of Mexico. NOAA Technical

Memorandum NMFS-SEFC-145, 25 p. plus 3 figures and 3 tables.

Wheaton, F. W. 1977. Aquacultural engineering. Wiley-Interscience,

John Wiley and Sons, New York, N.Y., 708 pp.

Wibbels, Thane. 1984. Orientation characteristics of immature Kemp's

ridley sea turtles, Lepidochelys kempi. NOAA Technical Memorandum

33
NMFS-SEFC-131, 67 p. (based on a Master of Science Thesis,

University of Houston, Texas, Texas, July, 1982).

Wood,J. R. and F. E. Wood. 1984. Captive breeding of the Kemp's

ridley, Marine Turtle Newsletter, No. 30, p. 12.

34
Table 1. Summary of head start results for Kemp's ridley sea turtle year-classes, 1978-1983.

Number of

turtles

Number Number held back Number of Number of

of live of dead Mortality Number of for research!?/ tagged tagged

Year- hatchlings hatchlings during head start Percent and captive turtles Percent turtles Percent

class received received head starting survivors surviva& brood stock released release~ recovered£! recovered

1978 3,080 1 992 2,088 67.8 42 2,019.21 65.6 75 3.7

1979 1,843 3 315 1,530 83.0 166 1,345~/ 73.0 18 1.3

1980 1,815 7 84 1,731 95.4 0 1,723V 94.9 84 4.9

1981 1,864 1 225P 1,639 87.9 0 1,639 87.9 46 2.8

1982 1,524 0 171 1,353 88.8 28 1,325 86.9 149 11.2

1983 250 0 58 192 76.8 2 190 76.0 6 3.2

Total 10,376 12 1,845 8,533 82.2 238 8,241 79.4 378 4.6

~Based on number of hatchlings received alive.

EYThese were abnormal, injured or sick individuals that could not be released.

E/AS of February 1, 1985 •

.2127 turtles died in transit.

~17 turtles died in transit.

V8 turtles died in transit.

~/Included 2 turtles unaccounted for but presumed dead.
Table 2. Tag numbers and other information for head started, Kemp's ridley sea

turtles of the 1982 year-class distributed to five public marine

aquaria.

Clutch Oate "Living- Internal Flipper

Recipient No.~/ Shipped Tagged"E! TagE/ Code Tag Codes!Y'

Clearwater Marine 9 11-9-83 No (°1-2;°2-20) NNM 106/107

Science Center 9

II No (°1-2;°2-21) NNM 154/155

Clearwater, FL 12 " No (°1-2;°2-32) NNM 193

12 II No (°1-2;°2-33) NNM 251/252

10 II Yes (01-2;°2-34) NNM 330

Key West Municipal 6 11-9-83 No (°1-2;°2-68) NNL 902/903

Aquarium 7

II Yes (01-2;°2-64 ) NNM 026

Key West, FL 8

II No (°1-2;°2-40) NNM 406

11 " No (°1-2;°2-41) NNM 448

19 II No (°1-2;°2-42) NNM 576

Theater of the Sea 10 11-9-83 Yes (01-2;°2-36) NNM 353

Fr.2/ 7

II No (°1-2;°2-65) NNM 023/024 Islamorada,

6

II No (°1-2;°2-66) NNL 917

8

II Yes (°1-2;°2-37) NNM 375

5

II No (01-2;°2-69) NNL 682/683
Table 2. continued

Clutch Date "Living- Internal Flipper

Recipient NO.~/ Shipped Tagged"b/ TagE./ Code Tag codes5Y

Gulfarium 4 1-26-84 ·No (D1-2;D2-2) NNL 485/486

Ft. Walton 3 " Yes (D1-2;D2-5) NNL 437/438

Beach, FL (died 5-29-84)

3 " No (D1-2;D2

-8) NNL 297/298

4 " No (D1-2;D2-4) NNL 475/476

20 " No (D1-2;D2-9) NNM 994

Marine Life Park, 5 2-6-84 No (D1-2;D2-1) NNL 665-666

Inc. , Gulfport, MS 18 " Yes (D1-2;D2-1O) NNM 872

17 " No (D1-2;D2-16) NNM 835

16 " No (D1-2;D2-17) NNM 790

15 " Yes (D1-2;D2-18) NNM 703

~Used by NPS, PINS and NMFS Galveston Laboratory.

b/"Living tags" were placed on left costal skute 3 (LC3).

~/Manufactured by Northwest Marine Technology Inc., Shaw Island,

Washington.

d/Monel metal tags. A single number indicates a tag on the right front

flipper. Two numbers separated by a slash indicate double-tagging

(i.e., tags on left and right front flippers, respectively).

~/These turtles were transferred from Turtle Kraals, Key west, FL to

Theater of the Sea, Islamorada, FL on April 16, 1985.
Table 3. Specifications, uses, and suppliers of various equipment and material

used in the head start project, 1978-1984~.

Equipment Descriptio~/

Fiberglass raceways, basins,

digestion tanks, and seawater reservoirs

Positive air blowers,

2.7 HP (Cyclonair

#CH5)

Hydrogen peroxide (H202)

Injector pump (Pulsafeeder

#680-S-E)

Centrifugal pump, 2 HP

(Gorman-Rupp

#B0024FGF2A4)

Submersible sump pump

140 gal/min at 5 ft

head (Teel pump #3P650)

Plastic buckets, 11 qt

volume, 28 cm diameter,

22 cm depth

Quonset hut covers

(Loratex, uv treated)

Titanium immersion heaters

Hydrogen peroxide, 35%

Use of Equipment

Holding turtles, treating wastewater, and

storing seawater

Aeration of digestion

tanks

Wastewater discharge

effluent treatment

by injection

Seawater delivery

pumping untreated

wastewater

Containers for individual sea turtles

Tops for quonset huts

Heating seawater in

reservoirs

Treating sea turtle

wastewater discharge

Equipment Supplier~/

Red Ewald, Inc.

P. O. Box 519

Karnes City, TX 78118-0519

Rotron, Inc.

Mansfield, OH

FMC Corporation

Specialty Chemicals Div.

Buffalo, NY 14240

Pump & Power Equip. Co.

800 Harwin Dr., Suite 370

Houston, TX 77036

Granger, Inc.

7777 Parnell St.

Houston, TX

Lorna Plastics, Inc.

Fort Worth, TX

Farm Supply Co.

500 Clarkesville St.

Cornelia, GA 30351

Glo-Quartz Elec. Heater Co.

7074-7190 Maple St.

Mentor, OH 40051

Accron Chemical Dist.

4700 Blaffer

Houston, TX 77026

~/Mention of trade names or commercial products does not imply endorsement or

recommendation for use.
Table 4. Monthly wastewater discharge effluent quality variables.

Arithmetic Avera es Ran es

Year & Salinity, Salinity,

Month ppt pH t

1981

July N,T~ NT NT NT NT NT NT NT

August 30 7.3 16.0 52.2 29-30 7.2-7.5 12-22 34-64

September 29 7.7 18.2 40.6 25-38 7.2-8.0 10-28 30-58

October 28 7.9 12.1 29.1 27-29 7.8-8.1 7-21 19-35

November 24 7.4 9.2 25.0 23-26 7.3-7.6 7-14 15-51

December 26 6.6 11.9 27.8 26* 5.9-7.1 5-19 15-43

1982

January 26 7.2 9.5 30.5 25-28 7.1-7.3 7-11 20-38

February 28 7.2 15.2 29.5 28-30 7.1-7.3 7-16 22-36

March 27 7.7 15.2 31.3 22-29 7.5-8.0 12-24 21-52

April 24 7.4 12.9 36.6 23-25 7.2-7.7 7-24 28-52

May 16 8.0 15.4 40.5 4-24 7.5-8.8 8-22 27-62

June NT NT NT NT NT NT NT NT

July 31 7.8 42.1 41.3 31-32 7.7-7.9 13-78 37-46

August 31 8.2 13.4 56.8 28-32 7.7-8.7 10-18 34-69

September 28 8.0 14.5 57.8 24-33 7.5-8.3 8-33 42-77

October 24 7.6 22.3 39.2 20-28 6.8-8.3 17-34 21-51

November 26 7.5 17.5 56.1 23-30 6.3-8.2 12-32 35-93

December 25 7.3 12.1 37.8 22-29 6.0-7.7 7-8 33-50

1983

January 23 7.5 9.7 47.4 22-24 7.3-8.4 9-10 39-51

February 24 7.5 10.0 46.0 22-25 7.2-8.4 9-10 36-51

March 28 8.3 13.4 47.4 22-29 7.2-7.9 8-25 38-54

April 28 7.8 8.8 40.6 26-30 7.7-7.9 8-10 31-54

May 26 8.5 11.2 30.6 9-36 7.7-8.1 9-16 24-62

June 16 7.9 11.0 56.6 13-20 7.9* 11* 51*

YNT = no turtles were in the facility; thus there was no discharge, so there

were no measurements taken.

E!TOC = total organic carbon.

S:/TSS = total suspended solids.

* = single value or no-variation.
Table 5. Ingredient~1 of the e~perimental diet£! manufactured by

central Soya and SubsidiarieS£/, Fort Wayne, Indiana.

Plant protein products

Animal protein products

Grain products

Processed grain by-products

Roughage products

Lecithin

Vitamin A supplement

Vitamin E supplement

Vitamin B12 supplement

Choline chloride

Niacin supplement

Calcium pantothenate

Folic acid

Riboflavin supplement

Menadione sodium bisulfite complex

O-activated animal sterol (source of vitamin 03)

Ascorbic acid

Biotin

Pyridoxine hydrochloride

Thiamine hydrochloride

Inositol

Methionine supplement

Ground limestone

Calcium phosphate

Salt and traces of:

Manganous oxide

Magnesium oxide

Ferrous sulfate

Calcium iodate

Ferrous carbonate

Cobalt carbonate

Copper oxide

Cooper sulfate

Zinc oxide

~/Quantities not available. Proprietary information of Central

Soya and Subsidiaries.

£/ory, floating pellets.

~/Mention of trade names or commercial products does not imply

endorsement or recommendation for use.
Table 6. Summary of Kemp's ridley sea turtle release sites, dates of

releases, numbers of turtles released, and tag series used for

six year-classes, 1978-1983.

Year- Imprint No.

class grou¢/ Release Site Release dates'£/ released Tag series~/

1978 PINS Sandy Key, FL 2-22-79 135 G----

PINS East Cape, FL~/ 2-22-79 52 G----

PINS ,East Cape, FL 2-28-79 1 13582

PINS East Cape, FL 2-28-79 166 G----

PINS Sandy Key, FL 3-5-79 172 G----

RN Homosassa, FL 5-8-79 751 G----, F----

PINS Homosassa, Fr2/ 5-8-79 628 G----, F----

PINS Padre Island, TX 7-7-79 112 G----, F----

RN Padre Island, TX 7-7-79 1 G0985

PINS Homosassa, FL 6-3-80 1 NNA260

1979 PINS Homosassa, FL 6-3-80 665 NNN---

(offshore)~/

RN Homosassa, FL 6-5-80 66 NNA---

(nearshore)

PINS Homosassa, FL 6-5-80 608 NNN---, NNA---

(nearshore)~/

PINS Padre Island, TX 6-2-81 5 K----

PINS Galveston, TX 9-28-81 1 J0096

1980 PINS Padre Island, TX 6-2-81 1,426 NNB---, K----

PINS Padre Island, TX 6-2-81 100 8001-8100 (Inconel)

RN Campeche, MX 3-3-81 197 NNB---, K----

1981 PINS Padre Island, TX 6-2-82 1,521 NNG---, NNH---

PINS Sabine Pass, TX 7-14-82 118 NNG---, NNH---

1982 PINS Padre & Mustang 6-7-83 1,159 NNL---, NNM---

Islands, TX

PINS Nueces Bay, TX 6-7-83 96 NNL---, NNM---

PINS Sabine Pass, TX 7-15-g3 69 NNL---, NNM---

PINS Mustang Island, TX 6-5-84 1 NNM428

1983 PINS Mustang Island, TX 6-5-84 172 NNQ---

RN Mustang Island, TX 6-5-84 18 NNQ---

Total 8,241

~PINS = "imprinted" at the Padre Island National Seashore;

RN ="imprinted" at Rancho Nuevo.

ElMonth-day-year.

E/Monel metal tags, unless noted otherwise. Each dash represents a numerical

digit from 0-9; actual numerical series are not given because they were

mixed. Details concerning the numerical series can be obtained from the

senior author of this paper.

~This release included turtles also tagged with radio-transmitters

(see Klima and McVey 1982; Wibbels 1984).
Table 7. Numbers of reported live and dead recoveries of head

started/tagged Kemp's ridley sea turtles of the 1978-1983

year-classes~/.

Year-class Live.£/ Dead Unknown..~/ Total

1978 63 8 4 75

1979 14 3 18

1980 46 17 21 84

1981 24 17 5 46

1982 94 48 7 149

1982 5 0 6

Totals

No. 246 94 38 378

% 65.1 24.9 10.0 100.0

~/AS of May 1, 1985.

'£/Returned to the environment.

E/rnformation not reported by person making the recovery.
Table 8. Frequencie~ of first and second choices by Kemp's ridley

sea turtles of pieces of marine shrimp abdominal muscle dyed

with food coloring.

Color of Food Color of Food Chosen First

Chosen Second Red Yellow Blue Green Non-dyed Totals

Red 171 44 15 25 19 274

Yellow 28 6 3 8 2 47

Blue 15 8 9 7 6 45

Green 37 8 5 12 5 67

Non-dyeaE./ 17 5 7 5 13 47

Totals 268 71 39 57 45 480

~/Number of times out of 480 total trials.

'£/Control.
Table 9. Observations~ on celestial orientation of hatchling, head started,

Kemp's ridley sea turtles at the Burke Baker Planetarium, Houston,

Texas.

Turtle 1D Azimuth Quadrant Total No. of

NumberE./ Date£! 1-900 91-1800 181-2700 271-3600 ObservationsY

1-H-5 7/28/83 1 1 3* 0 5

8/4/8~ 1 3* 1 0 5

8/11/83 1 0 1 3* 5

9/8/83 0 0 0 10* 10

Sub-total 3 4 5 13 25

2-H-6 7/28/83 1* 1 2 1 5

8/4/83 2* 0 3 0 5

8/11/83 0 0 5* 0 5

9/8/83 4 0 1 5* 10

9/16/83 (1st test) 3 0 1 6* 10

9/16/83 (2nd test) 1* 4 5 0 10

Sub-total 11 5 17 12 45

2-E-5 7/28/83 0 1 4* 0 5

8/4/83 3* 0 1 1 5

8/11/83 2 0 1* 2 5

9/8/83 1* 0 9 0 10

9/16/83 (1st test) 2 2 2 4* 10

9/16/83 (2nd test) 4* 2 0 4 10

Sub-total 12 5 17 11 45

l-D-4 7/28/83Y 0 0 0 0 5

8/4/83 0 2 3* 0 5

8/11/83 0 3* 2 0 5

9/8/83 1* 4 2 3 10

Sub-total 1 9 7 3 25

Total 27 23 46 39 140

~/Observations on the location of a hatchling were taken immediately at

1-minute intervals. An astrisk (*) marks the quadrant to which the

hatchling first moved after being released from the cage. The numbers

in the table indicate the number of times a hatchling was observed in a

given quadrant.

E./Raceway-bucket row-bucket column.

£IMonth/day/year.

YDoes not include the initial observation after release of the hatchling.

!:/This turtle displayed no directional response for the five minutes of

observation, but stayed in the center of the basin.
1'able 10. Results of a "living-tag" experiment in which carapace/plastron tissue grafts were applied by experienced and inexperienced taggers to

yearlings of the 1982 year-class of head started, Kemp's ridley sea turtles.

Type of Mark Ma"rk Location on Scute

Disc Gouge Subtotal % Lost Central Posterior Anterior Subtotal Percent Lost

Percent

"('agger Tagged lost Tagged lost Tagged lost Disc Gouqe Tagged lost Tagged lost Tagged lost Tagged lost lost Central Post. Ant.

Inexperienced

Tagger 1 54 29 54 28 108 57 53.7 51.8 36 19 36 20 36 18 108 57 52.8 52.8 55.6 50.0

Inexperienced

Tagger 2 54 4 54 0 108 4 7.4 0.0 36 36 36 2 108 4 3.7 2.8 5.6 3.7

Experienced 54 22 54 14 108 36 40.7 25.9 36 13 36 13 36 11 108 36 33.3 36.1 30.6 33.3

Tagger 1

Experienced 54 4 54 5 108 9 7.4 9.2 36 6 36 2 36 108 9 8.3 5.6 2.8 8.3

Tagger 2

Totals }:Io. 216 59 216 47 432 106 27.3 21.8 144 38 144 36 144 32 432 106 24.5 25.0 22.2 24.5
Figure 1. Average weight versus elapsed time for head started, Kemp's

ridley sea turtles of the 1978-1983 year-classes.
0.50

YEAR-CLASS

o 1978

o 1979

A 1980

• 1981

+ 1982

• 1983

INTERVAL

20 DAYS

11

I I

JUL

I

MAY

I

MAR

I

JAN

I

NOV

ELAPSED TIME (DAYS)

I

SEP

I

JUL

19

1.00

o

0.25

~ 0.75

4:

a:

Cl

o

..J

-~
Figure 2. Location of the sea turtle head start research facilities

(X) at the NMFS Galveston Laboratory.
DALLAS

I

GALVESTON ISLAND

PELICAN ISLAND

HOUSTON •

AUSTIN

•

•• co

..

co

'0

..

C?

It)

.......

.... ...

TURTLE HEAD START FACILITY

Figure 2. Location of the sea turtle head start research facilities

(X) at the NMFS Galveston Laboratory.
Figure 3. Insulated fiberglass seawater reservoirs supported by 6-in

thick, steel-reinforced, concrete pads.
Figure 4. Damage by Hurricane Alicia to the sea turtle head start

research facilities in August 1983.

A. Damage to facility, looking east; note upturned reservoir inside greenhouse frame structure.

B. Damage to wastewater treatment area and west digestion

tank; east digestion tank was totally destroyed.
Figure 5. Sea turtle head start research facilities after reconstruction following Hurricane Alicia in August 1983.
Figure 6. Raceway system used in rearing the 1978-1983 year-classes

of Kemp's ridley sea turtles.
Figure 7. The raceway standpipe system that prevents overfilling of

the raceways and allows quick drainage.
Figure 8. The bucket system used to isolate individual Kemp's ridley

sea turtles to prevent aggressive attacks.
Figure 9. The standing basin unit (in background) consisting of 120

hemispherical basins.
Figure 10. The sick-bay (isolation and treatment) unit consisting of

86 hemispherical basins.
Figure 11. Fiberglass wastewater-digestion tank and air-blower

house. The air-blower is elevated to prevent backsiphoning of wastewater into the blower.
Figure 12. Hydrogen peroxide (H202) injecter system. H202 is

injected into the wastewater effluent to achieve a

concentration of 60-100 ppm in the effluent as it

is being discharged.
Figure 13. Schematic diagram of tanks and plumbing used in the

wastewater treatment system. (1) wastewater discharge

pipe coming from raceway system; (2) in-ground sump pit;

(3) sump-pit overflow discharge line; (4) sump-pit pumps;

(5) sump-pit discharge lines; (6) aerated digestion tank

no. 1; (7) aerated digestion tank no. 2; (8) treated

wastewater discharge lines; (9) hydrogen peroxide

injector; (10) wastewater discharge sampling tap; (11)

discharge line to storm sewer; (12) bottom sludge drain;

(13) bottom sludge drain; (14) sludge discharge line; (15)

sludge discharge line; (16) domestic sewer line; (17)

freshwater flush line; (18) air-blower house; (19) bottom

air-blower pipe; (20) bottom air-blower pipe.
TO STORM SEWER 3

8

-11

10

5

13

1

(f)

>-

c(

=:

w

o

c(

a:

~

o

a:

u.

1 DISCHARGE LINE

2 SUMP PIT

3 OVERFLOW LINE

4 PUMP

5 DIGESTION TANK

6 TREATED-WASTE'

WATER LINE

7 HYDROGEN PEROXIDE

INJECTOR

8 SAMPLING TAP

9 BOTTOM SLUDGE DRAIN

10 SLUDGE DISCHARGE LINE

11 DOMESTIC SEWER LINE

12 FRESHWATER FLUSH TANK

13 AIR-BLOWER HOUSE

14 AIR-BLOWER LINE
Figure 14. Bucket code scheme for eighteen rows (lettered A-R, from

south to north) and six columns (numbered 1-6, from east

to west) of buckets, for a total of 108 buckets per

raceway. X marks bucket location G-4.
1 2 3 4 5 6

A 000000

B 000000

C 000000

D 000000

E 000000

F 000000

G 0000000

H 000000

1000000

J 000000

K 000000

L 000000

M 000000

N 000000

o 000000

p 000000

Q 000000

R 000000
"

Figure 15. A double-tagged, Kemp's ridley sea turtle. Tags are monel

metal, Hasco type, flipper tags.
Figure 16. Transportation box for tagged Kemp's ridley sea turtles.

Normally, eight tagged turtles (two horizontal layers of

four each) are transported in this manner to the release

site.
Figure 17. Food holder for food color preference study.
::u"m

" ." o m o ::u C m

::r:Z

o0

rm C (J) m -f

::u c:C

-<
Figure 18. Average concentrations (ppm) of (TOp) phosphate-phosphorus

(in phosphorus equivalents; P04-P) and (Bottom) ammoniumnitrogen (in nitrogen equivalents; NH4-N) in three

raceways (1,8, and 14) 48 hr after refilling (dashed

line) and immediately after refilling with clean seawater

(solid line).
- - - - 48 HOURS

- INITIAL

-:;:

0.

0.

-0.

I

'll:t

o

0.

.0

-:i

0.

0.

-Z

I

'll:t

:J:

z 0.4

/

/

/

,..., /

/ , /

;tJ' •••••••••••..• /

/\ /';tJ'"' /' / " '..J

" I

,/

.J

I

A I

/ \ I

I \

I \ A. I

\ '\ I

\ / \ I

\ I \ I

\ I \ I

\ I \ I

\ I \.------' \ I

\ .-- .

\.. --- - --J

1982

0.2

.0

22 24 29 31

AUG

5 7 12 14

SEP

19 21
Figure 19. Concentration (~g atoms per liter) of ammonium nitrogen

(in nitrogen equivalents; NH4-N) in a raceway containing

Kemp's ridley sea turtles and seawater treated by a

rotating, biological contactor (.) and in an untreated

(control) raceway (0). The test raceway contained 102

Kemp's ridley sea turtles averaging 723 g in weight, and

the control contained 108 averaging 740 g in weight.
o

• RACEWAY 11

1600L o RACEWAY 12

-.J

...•....••

en

:E 1400 0

to-

«

I

C)

.5 1200

z

0

-to-

« 1000

a:

toZ

IJJ

0 800 z

0

0

Z

IJJ 600 C)

0

a:

toz

- 400

:E

:::»

-

z

0 200 :E

:E

«

25 5 10 15 20 25 30 5

APR MAY JUN

ELAPSED TIME (DAYS)
Figure 20. Diagram of location of implanted, binary-coded tag in the

right front flipper of a head started Kemp's ridley sea

turtle.
MM~

Figure 20. Diaqr.~ of locat~on of implanted, binary·coded tag in the

right front flipper of a head started Kemp's ridley sea

.turtle.
Figure 21. Arithmetic average weight (kg) versus elapsed time for

ten head started Kemp's ridley sea turtles of the 1978

year-class held in captivity at Sea-Arama Marineworld,

Galveston, Texas.
25.0

20.0

en

:IE .

<C 15.0

a:

"

o 10.0 ...J

-~

o

2/14/80

300 600 900 1200 1500 1800

3/6/84

ELAPSED TIME (DAYS)
Figure 22. Sea turtle head start research facilities at the NMFS

Galveston Laboratory: (A) 1978-1981~ (B) 1981-1983~ (C)

1983-1984.
SEA TURTLE HEAD START FACILITY

A.1978-1981

8138888888

20 28 027 20

19

10 10 0111 12 13 14.15

37

250 [ill

e@@@ 35

17 18

1[38888 8

B.1981-1983

35

12 17 18

1/

29 SUMP PIT

30-31 DIGESTION TANKS

32 FRESHWATER RINSE TANK

33-34 10,OOO-GAL RESERVOI RS

35 SICK BA Y BASINS

36 TOOL HOUSE

37 QUONSET HUT

38 STORAGE

1 1

28

37

lo@@,.,o @ <8~31415 37

25

088888888 10

1-16 RACEWAYS

17-18 STANDING BASINS

19 6-FT DIA CIRCULAR TANKS

20 SIDE BASINS

21-24 7500-GAL RESERVOIRS

25-26 PUMPS

27 AIR BLOWER

28 DRAIN TROUGH

C.1983-1984
APPENDICES
Appendix Table A1. Results of food color preference study on Kemp's ridley sea

turtles: first trial from April 5-14, 1983. R = red, Y = yellow,

B = blue, G = green and C = undyed (control).

Test

Sequence

Turtle identification number~/ 1

First Choice

2 3 4 1

Second Choice

2 3 4

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

7-D-2

7-0-6

7-E-4

7-K-2

7-L-4

7-N-1

7-P-4

7-P-6

8-C-5

8-0-3

13-C-1

13-C-5

13-C-6

13-0-4

13-E-1

13-F-3

13-K-2

13-L-3

13-L-5

13-M-6

Y R R R

R R G R

G Y Y G

R R C R

R R C Y

eRR R

R R' R R

R R R G

Y R C C

Y Y R Y

R B Y Y

R R Y R

B R Y Y

Y R B R

R R C Y

R B Y R

R R C R

R Y B R

R R C Y

R Y G R

Y B

R G

C G

R R

R R

C Y

B G

B Y

G R

B Y

B C

R B

Y Y

R Y

R G

Y Y

Y R

R R

R C

C R

R R

Y R

G B

B R

R R

Y C

R Y

RG

R R

R R

R R

R B

R R

C R

R C

Y G

R R

C B

R R

R B

Y G

R R

R B

R R

R C

B Y

R R

B R

Y R

R R

R R

R R

R Y

Y Y

R R

R R

R R

R Y

R R

R R

Y Y

G R

G G

B R

C R

R B

R R

C C

Y R

C R

Y R

Y R

C R

C R

R R

Y Y

C R

C R

Y R

G Y

R G

G Y

B R

R R

G R

R R

R C

R B

Y C

Y C

Y R

R B

B Y

Y Y

R Y

Y Y

Y R

R R

Y R

R R

R R

R R

R B

B R

R R

C G

B Y

B B

B R

R R

R R

R R

C R

Y G

Y G

Y B

R R

C B

R B

R R

~Raceway no.-Row letter-Column number.
Appendix Table A2. Results of food color preference study on Kempts ridley sea

turtles: second trial from April 26 to May 5, 1983. R = red,

Y = yellow, B = blue, G = green and C = undyed (control).

Test

Sequence

Turtle identification number~1 1First Choice

2 3 4 1

Second Choice

2 3 4

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

10-A-4

10-B-4

10-C-1

10-C-2

10-c-5

10-E-4

10-E-6

10-F-1

10-G-2

10-G-3

3-J-2

8-J-3

8-J-6

8-K-2

8-K-4

8-L-1

8-M-4

8-0-2

8-P-1

8-P-6

Y R C Y

R G B R

Y R B R

C C B R

C G R Y

B Y R R

Y RB R

R R G C

C R R R

R R R R

Y R R R

Y G R C

Y R R R

R C R G

B R R B

G R C R

R R C B

G Y Y C

R R R R

G B G G

R R

Y R

G R

B B

Y R

RG

R Y

B G

R R

R B

C R

Y R

R R

R R

Y R

Y B

Y B

G R

R G

RR

B R

R R

R Y

R R

R G

R B

R R

R R

Y G

B G

R R

B R

B R

R R

R R

R R

R R

R R

R R

R R

R R

y y

R Y

B C

C G

G R

C R

R R

C R

R R

R R

Y Y

Y Y

R R

R R

R Y

Y R

R R

R R

R B

R R

R R

Y Y

Y C

R G

Y B

R Y

R R

R R

R R

R R

R B

R R

R C

C R

R R

Y B

C R

R Y

B R

B 8

R R

B B

R R

R R

B R

R R

R R

B R

B R

G G

R B

R C

B R

Y B

B G

B G

B G

G Y

B R

Y Y

R G

R B

B R

G B

8 G

B B

B R

B G

C R

R R

Y R

R R

R R

R R

R R

R R

R R

R R

R R

~/Raceway no.-Row letter-Column number.
Appendix Table A3. Results of food color preference study on Kemp's ridley sea

turtles: third trial from May 12-19, 1983. R = red, Y = yellow,

B = blue, G = green and C = undyed (control).

Test

Sequence

Turtle identification number~/ 1

First Choice

2 3 4 1

Second Choice

2 3 4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

13-N-S

13-0-2

13-0-6

13-Q-2

13-Q-6

13-R-1

14-A-3

14-A-5

14-0-2

14-F-2

14-H-4

14-H-6

14-J-l

14-J-3

14-K-l

14-K-5

14-L-l

14-M-3

14-N-2

14-0-2

R R

G R

R Y

C R

C R

R R

Y Y

R R

G Y

R Y

R R

R C

R Y

R B

G G

R Y

R Y

B R

R R

G R

B B

R R

R R

RR

R R

R R

C C

R R

R R

R R

R R

B R

R Y

RG

R R

R B

R R

R R

R R

R G

C R

Y R

R G

G R

Y R

R R

R C

R R

R R

R R

R C

R B

Y G

R C

R G

R G

B R

RC

RC

Y R

B R

R Y

R R

R R

R Y

R R

B C

R R

C C

Y Y

C R

C B

C Y

R R

R R

R R

R R

R R

R R

B R

R Y

R R

R R

C R

G R

R G

Y R

R B

R R

R R

R B

R G

R R

R R

C R

R R

R B

B B

R B

B R

R R

G R

R B

G G

G R

R B

G R

R B

R C

B R

R B

R C

B G

B R

B Y

C R

C B

C B

R B

B B

R R

G R

RG

R R

B R

R R

B R

R R

R R

Y R

R C

Y Y

B G

C R

R G

B Y

R R

R C

R C

R R

G B

R R

R R

R C

R R

RC

R R

B R

CR

Y R

C C

Y Y

C R

8 R

R B

R R

R B

R C

C R

R R

~/Raceway no.-Row letter-Column number.
Appendix B. Procedures, reagents, and standards used for seawater

chemistry analyses.

----------------------------------------------------------------------

A. Nitrate (N01-N)

Nitrat~, the most completely oxidized state of nitrogen in water,

is determined by the Brucine method (Jenkins and Medsker 1974). It

involves the reaction of N03-N with brucine sulphate in strong (13N)

sulfuric acid medium. Nitrite interference is eliminated by the addition of sulphanilic acid. Salinity interference is eliminated by

increasing the salinity of all samples and standards to a level above

which the effects of salinity are stable. Uneven heating, or, different temperature of samples produce erratic results. Therefore, all

samples (and standards) must be treated identically.

Reagents

NaCI solution - a saturated solution.

Sulfuric acid - add 5 parts (v.)H2S04 to 1.25 parts (v.)

distilled H20. Cool before using.

Brucine reagent - dissolve 1 g brucine sulphate and 0.1 g of

sulphanilic acid in 70 ml of hot distilled H20. Add 3 ml

concentrated HCl and make up to 100 ml with de-ionized water.

Procedures

1. Place 10 ml of sample in a test tube.

2. place test tubes in a rack submerged in icewater, the depth

of which should be sufficient to cover the portion of the

test tubes containing the samples and reagents.

3. Add 2 ml of NaCl reagent to each sample and mix by swirling.

4. Carefully pour 10 ml H2S04 reagent into each sample. Incline

each sample test tube for this addition.

5. Add 0.5 ml Brucine reagent to each sample.

6. Mix each sample by pouring back and forth one time in a

mixing tube and replace in ice water.

7. Remove tubes from icewater and place in a boiling-water bath
for 20 min (a yellow color proportional to the concentration

of N03-N in the sample develops during this time).

8. Stabilize color density by cooling samples in the icewater

bath to approximately room temperature.

9. Determine sample optical density at 410 mu in a spectrophotometer, or in a filter type electrophotometer using the

latter's 425 (purple) filter.

10. Compare sample optical density to standard optical density to

calculate concentration.

B. Nitrite N02-N)

The procedure used is that of Shinn (1941) as applied to seawater

by Bendschneider and Robinson (1952). The procedure is based on the

reaction of nitrite in saltwater with sulphanilamide in an acid solution. The resulting diazo-compound reacts with

N-(1-naphthyl)-ethylenediamine to form a highly colored (red) azo

dye.

Reagents

Sulfanilimide solution - dissolve 5 g of sulfanilimide in a

mixture of 50 ml of concentrated HCL acid and about 300 ml

of distilled water.

N-(1-naphthyl)-ethylenediamine dihydrochloride solution -

dissolve 0.5 g of the dihydrochloride in 500 ml of distilled

water. Store in a brown bottle.

Procedures

1. Place 25 ml of sample in a 50 ml flask.

2. Add 0.5 ml of sulfanilimide reagent and mix.

3. After at least 2 min add 0.5 ml of ethylenediamine reagent

and mix.

4. After no less than 10 min and no more than 120 min measure

the optical density of the samples in a spectrophotometer at

a wavelength of 543 mu, or in a filter-type electrophotometer
equipped with a green 525 mu-filter.

5. Compare sample optical density to standard calibration curve

to convert to concentration units.

C. Ammonium-Nitrogen (NH4-N)

We used the phenol-hypochlorite method of Solorzano (1969) as

described by Strickland and Parsons (1972). The procedure involves

treating samples in an alkaline citrate medium with sodium hypochlorite and phenol in the presence of sodium nitroferricyanide

(nitroprusside) that acts as a catalyzer. The intensity of the blue

indophenol that develops is an indication of the concentration of

NH4-N in the sample.

Reagents

Phenol solution - dissolve 20 g of crystalline reagent grade

phenol in 200 ml of 95% v/v ethyl alcohol.

Sodium nitroferricyanide solution-dissolve 1.0 g of sodium

ferricyanite in 200 ml of de-ionized water. Store in an

amber bottle.

Alkaline reagent - dissolve 100 g of sodium citrate and 5 g of

sodium hydroxide (analytical grade) in 500 ml of de-ionized

water.

Sodium hypochlorite reagent - use Clorox.

oxidizing solution - mix 100 ml of alkaline reagent and 25 ml

of clorox. Keep this solution stoppered while not in use.

Prepare fresh every day.

Procedures

1. Add 25 ml of sample to a 50 ml erlenmyer flask.

2. Add ml of phenol reagent to each sample and mix.

3. Add 1 ml of sodium nitroferricyanide reagent to each sample

and mix.

4. Add 2.5 ml of oxidizing reagent to each sample and mix.

5. Allow flasks to stand at room temperature for at least 1 hour
(color produced is stable for 24 hours).

6. Determine the optical density of the blue color in a

spectrophotometer at 640 mu, or in an electrophotometer with

a red filter.

7. Compare sample optical density to standard calibration curve

to convert to concentration units.

D. Phosphate-Phosphorus (P04-P)

The procedure used to determine P04-P in seawater is that of

Murphy and Riley (1962). It is also included in Strickland and

Parsons (1962). The procedure is based on the reaction of phosphate

in seawater with a composite reagent containing molybdic acid, ascorbic acid, and trivalent antimony.

Reagents

Ammonium molybdate solution - dissolve 15 g of reagent grade

ammonium molybdate [(NH4)6 M07024.4H20] in 500 ml of deionized water. Store in plastic bottle out of direct

sunlight.

Sulfuric acid solution - add 150 ml concentrated reagent grade

sulfuric acid to 900 ml of de-ionized water. Allow the solution to cool and store in a glass bottle.

Ascorbic acid solution - dissolve 27 g of reagent grade ascorbic

acid in 500 ml of de-ionized water. Store (frozen solid) in

a plastic bottle in a freezer. Thaw for use and refreeze at

once.

Potassium antimonyl-tartrate solution - dissolve 0.34 g of

potassium antimonyl-tartrate (tartar emetic) in 250 ml of

de-ionized water. Store in a glass, or plastic bottle.
Mixed reagent

Mix together 100 ml of the molbydate solution, 250 ml of the

sulfuric acid solution, 100 ml of the ascorbic acid solution

and 50 ml of the potassium antimonyl-tartrate solution.

Prepare this reagent for use daily and discard any excess. Do

not store longer than 6 hours.

Procedures

1. Add 25 ml of sample to a 50 ml Erlenmyer flask.

2. Add 2.5 ml of the mixed reagent.

3. Mix by swirling the flask.

4. After at least 5 minutes measure the optical density of the

blue color in a suitable photometer, or at 750 mu in a filtertype electrophotometer.

5. Compare sample optical density to standard calibration curve

to convert to concentration units.

E. Laboratory water for analyses

The de-ionized water used in our laboratory is prepared by passing

tap water, that varies from 200 to 800 ppm total solids, through a

mixed bed de-ionizer (which is supplied, and replaced as needed, by a

private contractor). Our distilled water is prepared by distilling

the de-ionized water with an all Pyrex still. Comparing blank values

showed that distillation of the de-ionized water did not improve its

quality as far as our chemical analyses were concerned. Therefore,

whenever distilled water is suggested in the analytical procedures,

de-ionized water may be used provided it is produced from a mixed bed

de-ionizer of the type that removes virtually all of the positive and

negative electrolytes.
F. Preparation of standards

Reagents used for preparing calibration standards must be readily

soluble, reagent-grade chemicals, dried before use. To minimize contamination, a bottle of each is set aside for standardization purposes

only. The following chemicals are used most frequently for calibration standardization at this laboratory:

1- Sodium nitrate (NaN03) for N03-N

2. Sodium nitrite (NaN02) for N02-N

3. Potassium acid phosphate (KH2P04) for P04-P

4. Ammonium chloride (NH4Cl) for NH4-N
Procedures:

1. Weigh a small amount of the desired chemical to the nearest

mg. For convenience, the weight should be somewhere in the

range of 0.1 g to 0.5 g.

2. Di9so1ve the weighed chemical into 1000x ml with de-ionized

water, where x is numerically equal to the gram weight of the

chemical. For example, if 0.235 g is the weight, it should be

dissolved in 235 ml of water. When the chemical is completely

dissolved and the solution thoroughly mixed (solution A), 1 ml

will contain 1 mg of the chemical. solution A is used to prepare the working standard from which calibration standards are

prepared. Assume a series of N02-N standards having a final

volume of 25 ml are desired that will increase in increments

of 1 u at N02-N/l; i.e., 1 ml standard + 24 ml H20 = 1 ug at

Nil; 2 ml standard + 23 ml H20 = 2 ug at Nil, etc. If solution A were used undiluted, 1 ml diluted to 25 ml would contain 580 ug at N02-N/I. The working standard solution,

therefore, is prepared by diluting 1 ml of solution A to

580 mI.

Appendix B Literature Cited

Bendschneider, K. and P. J. Robinson. 1952. A new spectrophotometic

method for the determinations of nitrite in seawater. J. Mar.

Res. 11:87-96.

Jenkins, D. and L. Medsher. 1974. Brucine method for determination

of nitrate in ocean, estuarine, and freshwater. Anal. Chem., Vol.

36, No.3 pp. 610-612).

Murphy, S. and J. L. Riely. 1962. A modified single solution method

for the determination of phosphate in natural waters. Anal.

Chem. Acta 27:31-36.
Shinn, M. B. 1941. A coloriometric method for the determination

of nitrite. Ind. Eng. Chern. [Anal. Ed.] 13(33).

Solorzano, L. 1969. Determination of ammonia in natural waters by

the phenolhypochlorite method. Limnol. and Oceanogr. 14:799.

Strickland, J. D. H. and T. R. Parsons. 1972. A practical handbook

of seawater analyses. Fish. Res. Bd. of Canada, Catalogue No. FS

94-167.