cine solution for 12–24 h (Kligerman and Bloom 1977). Adult Herpetological Review, 2005, 36(1), 45–47.
frogs, on the other hand, will need to be injected with a small 2005 by Society for the Study of Amphibians and Reptiles amount of colchicine (1 mg/ml deionized water) using a 23-gauge PIT Tag Retention in Trachemys and Pseudemys
needle (1.9 cm in length). To ensure circulation throughout thetorso the colchicine solution should be injected through the skinof the dorsum and into the abdomen at a depth of ca. 3 mm. A frog ALIYE L. RUNYAN
5 cm in length, such as Gray Treefrogs, should be injected with PETER A. MEYLAN
ca. 0.5 cc of the solution. Frogs of larger or smaller size should be Eckerd College, St. Petersburg, Florida 33711, USA injected with amounts scaled up or down relative to the amount used for adult Gray Treefrogs. Once injected, adult frogs should be placed into a container with a moist paper towel at 30°C for10–12 h (Bogart 1967).
Passive integrated transponders are small microprocessors en- Once the specimens (either tadpole or adult frog) have been cased in glass that transmit a unique identification number to an subjected to the colchicine solution for the correct time, they should electronic reader when the tag is activated by the reader with a be sacrificed by the application of Orajel® to the top of the head low frequency radio signal at close range (Balazs 1999). PIT tags (Altig 1980). Once sacrificed, the specimens can be dissected and were originally developed to census fish populations (Boarman et tissue samples may be taken from various locations on the speci- al. 1998) and to track fish migration (Prentice et al. 1987). How- men. Leg buds, gills, and intestinal tissue samples were the most ever, their use has become widespread in the identification and successful from tadpoles and intestinal tissue worked best for adult monitoring of individuals of other vertebrate groups. PIT tags have frogs. The tissue sample should be no longer than 0.5 cm in length been successfully used in studies of mammals, such as otters, rats, and should be placed in a small vial containing about 10 times bats, mice, ferrets (Elbin and Burger 1994), and voles (Harper and their volume of 0.4% KCl hypotonic solution and should be al- Batzli 1996). PIT tagging has been used on a variety of amphib- lowed to sit for 20–30 min (Kligerman and Bloom 1977). Trans- ians and reptiles, including lizards, alligators, frogs, and toads fer the tissue into 2–3 changes of fixative (3:1 ethanol:acetic acid (Elbin and Burger 1994), neonatal snakes (Keck 1994), caiman solution) for ca. 30 min each time (Kligerman and Bloom 1977).
hatchlings (Dixon and Yanosky 1993), and rattlesnakes Mount chromosomes on slides using heat fixation. This can be (MacGregor and Reinert 2001). In freshwater turtles (Elbin and accomplished by heating blank slides 46–48°C. The tissue sample Burger 1994; Buhlmann and Tuberville 1998), and sea turtles should then be placed into a separate vial containing 100 µl 50% (Balazs 1999; McDonald and Dutton 1996), PIT tags have also acetic acid (Kligerman and Bloom 1977). Tap the vial for 60–90 been proven successful. In a study of PIT tag utility, Camper and sec to break apart the tissue and form a cell suspension. Using a Dixon (1988) tagged 24 individuals of eight different species of pipetman, draw the cell suspension into the tip and expel it onto freshwater turtles, including eight Trachemys scripta elegans. the slide. Quickly withdraw the suspension back into the pipet tip, PIT tag effectiveness has been discussed in a number of differ- leaving as little liquid on the slide as possible.
ent studies, including one on the Pine Snake (Elbin and Burger Stain chromosomes with 4% Geimsa made up in 0.01 M phos- 1994), and another on a large group of Desert Tortoises (Boarman phate buffer at pH 7 for 10 min. Air-dry the slides and place in et al. 1998). Both of these studies came to the conclusion that PIT xylene for 10 min to remove excess water. Once dried, the slides tags were an efficient and reliable method of marking and track- can completed by using Permount and a cover slip over the fixed ing their respective subjects. There have been relatively few re- ports on tag retention in freshwater turtles, Buhlmann andTuberville (1998) being the only one with large sample sizes known Acknowledgments.—We thank P. Villani for assisting with digital im- to us. Balazs (1999) discussed some of the advantages and disad- ages of the chromosome spreads. This study was supported by two grants vantages of tagging sea turtles with PIT tags. Advantages of the from the Augustana Research Foundation, Augustana College: Bernsten tags include virtually no loss or damage over time from breakage Award to LG, and Faculty Research Grant to SBH.
or corrosion, which offers the possibility of long-term reliable re-tention in sea turtles. Disadvantages include migration within body tissue, inability to detect a tag without a reader, and overall highercost for the PIT tags (US $6.00 each) and reader (~ US $400) as ALTIG, R. 1980. A convenient killing agent for amphibians. Herpetol. Rev. compared to external tags. PIT tags appear to have no adverse BOGART, J. P. 1967. Chromosome of the South American amphibian fam- effects upon the freshwater turtles into which they are injected ily Ceratophridae with a reconsideration of the taxonomic status of Odontophrynus americanus. Can. J. Genet. Cytol. 9:531–542.
During the course of a study of the impact of the invasive turtle GOSNER, K. L. 1960. A simplified table for staging anuran embryos and Trachemys scripta elegans on native Pseudemys floridana and P. larvae with notes on identification. Herpetologica 16:183–190.
nelsoni populations in St. Petersburg, Florida, PIT tags were in- KLIGERMAN, A. D., AND S. E. BLOOM. 1977. Rapid chromosome prepara- jected into each captured turtle in order to recognize individuals tions from solid tissues of fishes. J. Fish. Res. Board Can. 34:266–269.
of all three species. Our goal here is to examine tag retention and WASSERMAN, A. O. 1970. Polyploidy in the common tree toad Hyla versi- tag movement in the turtles marked for that study and to report on color. Science 167:385–386.
the utility of a specific injection site between the pelvis and plas-tron. Furthermore, we assess the impact of PIT tags on femalereproduction.
Since April of 1998, retention and movement of PIT tags from the site of implantation in turtles of three species were studiedusing x-rays. Throughout the study, x-rays were taken to deter-mine if captured females were carrying eggs (Gibbons and Greene1979). This also allowed the presence and movement of PIT tagsto be observed. Most of our data are from adult female T. scriptaelegans; however, small samples of female Pseudemys floridanaand P. nelsoni were also tagged, recaptured, and x-rayed, as weresmall samples of males of all three species. Females were keptovernight to be x-rayed at a veterinarian’s office. Three males,two P. floridana and one T. s. elegans, were x-rayed to confirmthe location of the PIT tag after a number of years. Turtles werereleased into the ponds from which they were captured.
Turtles were captured using hoop traps (Nylon Net Co.) in six ponds on the Eckerd College campus, St. Petersburg, Florida (Emer2004; Hutchinson 1992). Upon first capture, each turtle was givena unique three-letter code that was drilled into their marginal scutes(Gibbons and Greene 1979), which was used to identify turtles incase of PIT tag loss. Beginning in April 1998, PIT tags were usedalong with scute drilling. Prior to injection, the turtle was placedon its carapace. The tag was injected with a 13-gauge needle intomuscles and connective tissue between the pelvis and plastron justlateral to the midline. This location was chosen because it is be-tween two plates of bone that move little relative to one another,subjecting the tag to the least disturbance possible. The needlewas inserted just through the skin into the tissue dorsal to thexiphiplastron and ventral to the ischium, lateral to the cloaca. Thetag was delivered into connective tissue and muscle by a plungerbuilt into the syringe. Care was taken to avoid any inadvertentlateral tail movement during insertion of the needle. When turtleswere recaptured, PIT tags were read using an AVID, Inc. scannerby placing the scanner along the interanal seam of the plastron.
FIG. 1. X-ray of male Pseudemys floridana showing the location of the The locations of the PIT tags were followed using x-rays in a PIT tag; located laterally and to the right of the midline.
total of eleven female T. s. elegans. In these turtles, the PIT tagsstayed in essentially the same location into which they were in-jected. These eleven females had been PIT tagged over a period of x-rayed. Two males were x-rayed at recapture; one had retained five months (April–June 1998). They were found to have retained the PIT for 36 months, and the other for 14 months between cap- the tags at subsequent captures, ranging from 30 days to 48 months tures (Fig. 1). The female was tagged and x-rayed on first capture, later. In two females, the PIT tags moved from one side of the but was not recaptured. In the turtles that were checked with the midline to the other over a period of 16 days to 16 months, respec- scanner but not x-rayed, tags could be read at subsequent recap- tively. Another female died during the course of the study of causes tures, spanning periods of 20–30 months. Only one of three P. not related to the tag implant; this female had lost a PIT tag and nelsoni has been tagged and subsequently recaptured. This adult had a second one injected. It is believed that this turtle lost the female had retained the PIT tag for 32 months.
first tag because it was placed too far posterior of the ideal injec- It appears that PIT tags are reliable if properly injected into the tion site and was lost through the injection site. The second tag location described above, and the tags do not get lost when placed was placed in the proper position and was retained for 45 months in connective tissue where very little movement occurs. In this until her death. At least two females developed, carried, and laid study, the tags did not appear to be more effective than carapacial clutches of eggs without affecting the position of the PIT tag, nor drilling. However, if tracking of large numbers of turtles were did the tag appear to interfere with the normal reproductive pro- necessary (Boarman et al. 1998), or for tagging of softshell turtles, cess. One male T.s. elegans was x-rayed 50 months after implan- PIT tags would be more efficient. Another consideration with the tation of the PIT, and the tag was found to be in the same location use of the PIT tag system is that while electronic readers cost on into which it was injected originally. Seventeen other males were average US $400, PIT tags remain expensive at US $6 per tag.
tagged, and retained their PIT tags without loss between captures This may affect the use of PIT tags in a study that monitors large spanning from 73 days to 35 months. All tags were readable at numbers of animals. The tags are an easier and quicker method of marking turtles than scute drilling, and while carapacial scute PIT tags in a total of eight Pseudemys floridana and three P. markings may grow back after one to two years in juvenile nelsoni were also monitored. Two female and six male P. floridana Pseudemys (Meylan, unpubl.), the PIT tags remain in place and were tagged, but only one female and two males were subsequently are readable for at least 50 months after implantation. The lifespan of a PIT tag is theoretically the lifetime of the animal; because aPIT tag is a passive marking system, as long as it remains inside HERPETOLOGICAL HUSBANDRY
the animal, it should be readable. The first successful PIT tag wasmade in the early 1990s, and is still working today, so the lifetime Herpetological Review, 2005, 36(1), 47–49.
is at least 14–15 yrs (AVID, Inc., pers. comm., 16 January 2004).
2005 by Society for the Study of Amphibians and Reptiles PIT tags are easier to read and provide a check on drilling or notch-ing, thus leading to fewer errors when large numbers of collabora- Notes on the Captive Husbandry of the King
tors collect data, or when large numbers of animals are used in a Cobra (Ophiophagus hannah) at the Centre for
Herpetology/Madras Crocodile Bank, India
Acknowledgments.—This study has been supported by Eckerd Col- lege. Many thanks to Diana Huestis, the Eckerd College Herpetology R. WHITAKER
P.O. Box 21, Tamil Nadu, Chengalpattu 603 002, India Club, and Drs. Richard Wilkes and Shawna Greene at Bay MooringsAnimal Hospital. Thanks also to Jim Dixon for reviewing and providing N. WHITAKER
Madras Crocodile Bank/Centre for Herpetology, P.O. Box 4 Mamallapuram BALAZS, G. H. 1999. Factors to consider in the tagging of sea turtles.
IUCN/SSC Marine Turtle Specialist Group, Publ. No. 4. 101–109.
Carmelaran Post, Sarjapur Rd., Karnataka, Bangalore 560 035, India BOARMAN, W. L., M. L. BEIGEL, G. C. GOODLETT, AND M. SAZAKI. 1998. A passive integrated transponder system for tracking animal movements.
Despite being such a large, conspicuous snake, the King Cobra (Ophiophagus hannah) was not described until 1836 (Cantor 1836).
BUHLMANN, K. A., AND T. D. TUBERVILLE. 1998. Use of Passive Integrated To date, little is known regarding its natural history and reproduc- Transponder (PIT) tags for marking small freshwater turtles. Chelon.
Conserv. Biol. 3:102–104.
tive biology. For a summary of field observations of reproductive CAMPER, J. D., AND J. R. DIXON. 1988. Evaluation of a microchip marking system for amphibians and reptiles. Texas Parks and Wildlife Depart- Examination of reproductive data reveals some interesting trends.
ment, Research Publication 7100-159:1–22.
In general, mating occurs in the post-winter warming period, egg DIXON, J. R., AND A. A. YANOSKY. 1993. A microchip marking system for development within the female occurs in the tropical summer, nest- identification of caiman hatchlings. Bull. Maryland Herpetol. Soc.
ing and egg laying coincide with pre-monsoon showers, and hatch- ing occurs at the peak of the monsoon. Captive snakes in locales ELBIN, S. B., AND J. BURGER. 1994. Implantable microchips for individual far removed from monsoon seasons remain faithful to this breed- identification in wild and captive populations. Wildl. Soc. Bull. 22:677– ing cycle. While the monsoon in Chennai (where no King Cobras EMER, S. 2004. Growth of a population of Trachemys scripta elegans at occur) is in the winter, reproductive behavior in our captive King Fox Pond, Eckerd College, Pinellas County, Florida. Herpetol. Rev.
Cobras followed the southwest monsoon cycle of southwestern GIBBONS, J. W., AND J. L. GREENE. 1979. X-ray photography: a technique In 1996, we established a conservation-oriented captive-breed- to determine reproductive patterns of freshwater turtles. Herpetologica ing program for King Cobras at the Madras Crocodile Bank/Cen- tre for Herpetology (MCB/CFH). The objectives were to (a) de- HARPER, S. J., AND G. O. BATZLI. 1996. Monitoring use of runways by velop methods and a protocol for the breeding of King Cobras in voles with passive integrated transponders. J. Mammal. 77:364–369.
India, and (b) to offer captive bred King Cobras to zoos in order to HUTCHINSON, A. M. 1992. A reproducing population of Trachemys scripta discourage the taking of animals from the wild. Herein, we report elegans in southern Pinellas County, Florida. Herpetol. Rev. 23:74–75.
KECK, M. B. 1994. Test for detrimental effects of PIT tags in neonatal husbandry and captive breeding information along with previously MACGREGOR, G. A., AND H. K. REINERT. 2001. The use of passive inte- Husbandry.—In 1996, MCB/CFH received seven (three males grated transponders (PIT tags) in snake foraging studies. Herpetol. Rev.
and four females) adult King Cobras; four of these were on breed- ing loan from Indian zoos and three had been seized by the Wild- MCDONALD, D. L., AND P. H. DUTTON. 1996. Use of PIT tags and photo life Department from snake collectors.
identification to revise remigration estimates of leatherback turtles A block of eight rooms was constructed of brick and mortar (Dermochelys coriacea) nesting in St. Croix, U.S. Virgin Islands, 1979– with light roofing. Each room measured 3.5 x 1.5 x 2 m (L x W x 1995. Chelon. Conserv. Biol. 2:148–152.
H). At the rear of each room (opposite the door) is a 100-mm air PRENTICE, E. F., T. A. FLAGG, AND S. MCCUTCHEON. 1987. A study to deter- mine the biological feasibility of a new fish tagging system, 1986– duct connected to an air conditioner, which from April to Septem- 1987. Annual Report of Research. U.S. Dept. Energy, Bonneville Power ber (1000–1600 h) creates a temperature gradient of about 6ºC, Administration, Division of Fish and Wildlife. 112 pp.
increasing to 30ºC towards the door. Outside the rooms, the ambi-ent temperature may reach 40ºC + during this period. Throughoutthe rest of the year (cooler months with lows of 18– 20ºC), a heat-ing pot containing a 25-watt bulb placed in an upturned mud pot issituated in the rear corner of each room. The surface temperatureof the exposed section of the pot averages 31ºC. Each room has a


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