Annals of Warsaw University of Life Sciences – SGGW Science No 52, 2013: 243–252 (Ann. Warsaw Univ. of Life Sci. – SGGW, Anim. Sci. 52, 2013)

Comparative behaviour analysis of some colubrids with reference to suitability of captive bred for reintroduction to natural habitat ANDRZEJ ĩYCZYēSKI, ZUZANNA NOWAK Department of and Animal Breeding, Warsaw University of Life Sciences – SGGW

Abstract: Comparative behaviour analysis of INTRODUCTION some colubrids with reference to suitability of captive bred snakes for reintroduction to natural Herpetofauna belongs to that most vul- habitat. A sample of newly hatched American colubrids, divided according to their level of nerable group of and those most domestication, understood herewith as the gen- endangered by anthropogenic pressure eration history in captivity, was tested in an open to their natural habitat. There have been ¿ eld test (OFT) for degree of activity and con¿ - many attempts of in situ conservation dence in open terrain and in confrontation with in- and some have proved quite effective dividuals of such as guttatus and Lampropeltis getula splendida as well as L. g. – for more information on this, see the californiae, in order to compare their antagonistic Materials of VI World Congress of Her- and feeding behaviour reactions. The cluster com- petology – Manaus 2008. Despite these parison of defence reactions revealed no effect of efforts, more and more trials are under- domestication (many generation in captivity) in taken for ex situ strategies. Traditionally the face of potential danger. Even albino phase , considered to be the form of this spe- this activity is the domain of universities cies most affected by captivity (altered genome), or ZOOs, which maintain certain species maintained their natural feeding response. The as gene banks – potential sources of ma- degree of activity in OFT conditions was seen to terial for reintroduction. In the case of decrease with the snakes’ age. Thus it was demon- snakes, there are quite a few examples strated that captive specimens bred in terrariums of attempts in this ¿ eld. During the years may be considered a gene bank for ex situ conser- vation strategy. However it is recommended that 1980–1986, some 34 specimens of cap- specimens as young as possible should be used tive bred Rocky Python Python sebae in reintroduction to natural habitat, as these will were released into the Great Fish River respond most faithfully to the pressure of natural Nature Reserve (Eastern Cape), but there selection, based on their behavioural variation not was no follow-up (Alexander and Marais supressed by apathy caused by captivity. 2007, Mattison 2013 – personal commu- Key words: Lampropeltis, Pantherophis, antago- nication). In the USA, ¿ ve AZA facilities nistic and feeding reactions, captive breeding, have created a consortium “Conservation ex situ conservation 244 A. ĩyczyĔski, Z. Nowak

Centre for Species Survival” undertak- though as early as 1975, John Coborn ing preparatory work for reintroduction organized the symposium “Conservation programs for various taxons, including and Captive and ”, snakes. The species of choice are to be where the case was made by many her- the San Francisco garter Thamno- petologists for and against captivity as an phis sirtalis tetrataenia and the Louisi- aid to the preservation of wild species, ana Pine Snake ruthveni (Mat- there is still much work to be done in tison 1995, Conway 2011, Mattison 2013 this ¿ eld. In any case, it must be admit- – personal communication). Godwin ted that the level of knowledge of her- et al. in 2008 presented a comprehen- petofauna husbandry is constantly grow- sive report on the captive propagation ing and additionally, the pure breeding of the critically endangered Eastern In- strategy according to locality of origin digo Snake Drymarchon couperi, which has become the ethical rule among the included suggestions concerning other most advanced keepers. As a result, in snake species. An initial scenario was many cases the numbers of certain spe- developed for supplementing the local cies in captivity exceed those ever to be population of Grey Banded seen in the wild and these can be treated Lampropeltis alterna by captive animals as pure gene banks. Their active popula- (Spanowicz and ĩyczyĔski 2002). Simi- tion, with a quite large effective number lar efforts were undertaken by àódĨ Uni- (Ne) is much greater than could be as- versity, in cooperation with àódĨ ZOO, sured by of¿ cial institutions limited by for a domestic (Polish) species – Smooth state budget conditions. But if we are to Snake Coronella austriaca (ZieliĔski treat captive animals as candidates for re- and Stanisáawski 2001, Stanisáawski stocking, we must be sure that these ful¿ l 2003). All these attempts led to the re- the demands of gene bank purity, as well lease of captive snakes into the wild, but as the demands with regard to sanitarian were not continued after the ¿ rst trial. status, in order to prevent introduction of Another attempt worth mentioning exotic pathogens to the natural habitat. was the successful reintroduction of the The captive material must also exhibit Antiguan Racer Alsophis antiguae (Dal- the natural degree of ¿ tness that can be try 1999 and 2006). In this project, the subjected to natural selection. reintroduced snakes were ¿ eld collected The aim of the present study is a com- on Great Island, tagged and released parative behavioural analysis of captive on small Rabbit Island. On the contrary bred snakes, which can be differentiated the results of reintroduction of Woma according to the level of their domestica- Python Aspidites ramsayi in Arid Recov- tion (understood herewith as the multi- ery Reserve in northern South Australia generation history in terrariums’ life). did not meet the short-term and medi- um-term success criteria mostly due to predator pressure (Moseby et al. 2011). MATERIALS AND METHODS Potential support for such work may The snakes used in this study were born come from private enthusiasts and keep- in Laboratory of Department of Genet- ers. This is a signi¿ cant new idea. Al- ics and Animal Breeding of WULS Comparative behaviour analysis of colubrids with... 245

– SGGW. They belonged to the genera The snakes were subjected to open Lampropeltis and Pantherophis. Au- ¿ eld test – OFT (Markowska 1979) scor- thors chose these taxons in view of their ing points for movement activity within popularity among keepers. Compared to the ¿ eld. Animals were placed within other genera, most of these snakes have a ring 90 × 90 cm, fenced with wooden a quite long generation history in captiv- walls. The surface of the ring consisted ity and thus constitute interesting materi- of squares 30 × 30 cm forming three al for an evaluation of the level of natural rows. The scores for activity were count- behaviour preserved. These may serve ed as follows: snake was getting one as a model for conclusions concerning point when slithered along one square other, potentially truly endangered spe- adjacent to the walls and 3 points for dis- cies, which are currently not stocked in tancing from the ring walls and entering collections. the square in the centre of the ¿ eld. Af- The material was divided into the fol- ter 5 min of observation the points were lowing groups: added up. 1. Ls × Lc – inter subspecies cross be- Later the snakes were confronted tween Sinaloan Lampro- with a young Corn Snake Pantherophis peltis triangulum sinaloe and Pueblan guttatus – a species preferring as Milk Snake Lampropeltis triangulum the natural diet, and a Desert Kingsnake campbelli – (parents from multigenera- Lampropeltis getula splendida – liv- tional captive breeding) – here – captive ing (as the whole genus Lampropeltis) (domesticated) type – 10 specimens; on lizards and other snakes as its most 2. Lpp × Lpk – inter subspecies cross be- typical food. The points scored and body tween Arizona Mountain Kingsnake weights of each snake tested were sub- Lampropeltis pyromelana pyromelana jected to one-way ANOVA. When nec- and Chihuahua Mountain Kingsnake essary the F-test was followed by D-test Lampropeltis pyromelana knoblochi to verify the signi¿ cance of differences – second generation in captivity – in- among the groups. Additional analysis of termediate type – 3 specimens; covariance with body weight as an oper- 3. Ege – Great Plains Pan- and variable was done to check whether therophis emoryi – from ¿ eld collected the sizes of snakes could affect their con- parents – wild type – 8 specimens; ¿ dence and thus the results of OFT test. 4. Ege × – interspecies cross be- During confrontations (lasting 5 min tween Great Plain Rat Snake Pan- each) with another snake, the following therophis emoryi (¿ eld collected) and reactions were noted on a Yes or No basis: Corn Snake Pantherophis guttatus lack of any reaction, strong tail rattling (amelanistic form – ¿ xed mutation in (loud and lasting longer than 5 s), weak multigenerational captivity) – inter- tail rattling (lasting less than 5 s), tossing mediate type – 6 specimens; movements, sudden retreat, avoidance 5. Eggalb × Eggan – Pantherophis gut- and attack. tatus – a crossbreed of two ¿ xed mu- In authors’ experience, the chosen/ tations – snow albinism and anery- /described reactions reÀ ect well the emo- trystic (black albinism) – “domestic” tional status of the snake during the test. type – 7 specimens. The probabilities of each reaction in two 246 A. ĩyczyĔski, Z. Nowak separate trials, for confrontations with only. As the snakes aged, the differences Corn Snake P. guttata and Desert King- between groups disappeared. snake L. g. splendida and combined, The score in the II observation is gen- were subjected to cluster analysis (Nei erally smaller – vide Table 1. This can be 1972) permitting to build dendrograms explained by increased apathy caused by of behavioural distance. captive conditions. In an additional analysis (not in- The ANOVA results for snakes body cluded in all experiments), authors used weights were signi¿ cant P ” 0.05. As Sinaloan Milk Snake Lampropeltis tri- the activity in OFT could be affected by angulum sinaloe – 4 specimens, Grey- snakes’ sizes we made the additional anal- -banded Kingsnake Lampropeltis alterna yses, correcting the analysis for points – 2 specimens, and scored in the ¿ rst observation by deviation Lampropeltis getula californiae (albino) from regression line for body weights. – 2 specimens. All of these were born The deviations from regression line a year later. These snakes were confront- (body weight as an operand variable) are ed with each other within their own spe- highly signi¿ cant (P ” 0.01) and con¿ rm cies and with albino Kingsnakes. that the signi¿ cant differences (P ” 0.05) in points scored in OFT (I observation – Table 1), were not caused by differences RESULTS in the sizes of specimens, but in reality The points scored in OFT in both obser- depended on group classi¿ cation. vations met the criteria of normal distri- Activity in the ¿ eld test was lower bution with mean values 7.7 and stand- during the II observation except in the ard deviation 5.4 and mean value 8.5 case of the group 5. Growth was differ- with standard deviation 5.5 for I and II ent in each group – in the pyromelana trial respectively. group, this was even a negative value. The differences between groups are The results for cluster analysis are signi¿ cant P ” 0.05 for the I observation shown in the dendrograms in Figures 1, 2, 3, 4 and 5. TABLE 1. Points scored in OFT and measurements of body weights during the I and II observations Scores in OFT Body weights [g] Groups N I Observation II Observation 1st Observation 2nd Observation x r V x r V x r V x r V Ls × Lc 10 7.9 ±0.9a 7.3 ±3.2 35.2 ±3.1A;E 36.4 ±3.3 Lpp × Lpk 3 3.3 ±1.8b;d 1.7 ±2.1 27.7 ±1.0B;F 26.3 ±3.3 Ege 8 10.9 ±2.5d;e 8.0 ±5.4 174.9 ±34.7A;B;C;D 185.1 ±37.0 Ege × Eggam 6 13.2 ±1.9a;b;c 10.3 ± 6.4 112.0 ±21.1C;E;F;G;H 141.9 ±21.3 Eggan × Eggalb 7 4.7 ±1.3c;e 7.6 ±1.9 40.5 ±2.9D;H 47.9 ±4.0 Letters (in pairs): a–e point out the signi¿ cant differences between means of groups for points scored during the ¿ rst trial – Duncan test; P < 0.05; Letters (in pairs): A–H show the signi¿ cant differences between means of groups for body weights dur- ing the ¿ rst trial – Duncan test; P < 0.05. Comparative behaviour analysis of colubrids with... 247

FIGURE 1. Comparison of group reactions during the I observation – confrontation with Pantherophis guttatus

FIGURE 2. Comparison of group reactions during the II observation – confrontation with Pantherophis guttatus

FIGURE 3. Comparison of group reactions during the I observation – confrontation with Lampropeltis getula splendida 248 A. ĩyczyĔski, Z. Nowak

FIGURE 4. Comparison of group reactions during the II observation – confrontation with Lampropeltis getula splendida

FIGURE 5. Comparison of group reactions – both types of confrontation grouped together

Confrontation with the relatively blood” as a factor differentiating the harmless Corn Snake Pantherophis gut- groups. tatus has not revealed any differences in During confrontation with their own reactions between groups, which could species (Greybanded Kingsnakes, Milk be attributed to either the level of domes- Snakes and California Kingsnakes), au- tication or the taxonomic position. thors observed some slight excitation In contrast to the I observation, dur- (avoidance, tail rattling) in the tested ing the II observation, played snakes. Confrontation with the albino an important part in sorting the snake Kingsnake resulted in an attack by the groups within clusters. This tendency re- latter showing the typical feeding re- mained constant during subsequent trials sponse. The Kingsnakes themselves did – vide Figures 3–5. not react to each other. Figures 3–5 show dendrograms that Following the attack, the trials were are practically identical with Figure 2. immediately halted, with no harm being There is no noticeable inÀ uence of do- caused to the snakes by the testing pro- mestication or the addition of “wild cedure. Comparative behaviour analysis of colubrids with... 249

DISCUSSION – probably due to apathy caused by cap- tive conditions. Analysis of the snakes’ activity in OFT Another source of information is pro- showed signi¿ cant differences only dur- vided by analysis of the dendrograms. ing the I observation (Table 1). Compari- The I confrontation, with the relatively son within genus Pantherophis revealed harmless Corn Snake (Fig. 1), showed that groups 3 and 4 did not differ signi¿ - a considerable variance in reactions be- cantly from each other but both deviate tween groups, which could not be attrib- from group 5 (the smallest specimen uted to either taxonomy or level of do- – Table 1) designated the most domes- mestication. The II confrontation, with ticated – the longest generation history a Corn Snake and the effect of exposing in captivity. Correction on regression the tested specimens to more danger- line for body weight showed that points ous stimuli i.e. Desert Kingsnake L. g. scored in OFT do not depend on body splendida, (Fig. 2–5) provided uniform weight. This result accords with the fact dendrograms arranged according to tax- that during the II observation, differences onomy and not the captivity status. between groups in terms of body weight Albino Kingsnakes tolerated each increased, but differences in activity other well, but in the presence of another (points scored in OFT) disappeared (vide species, manifested an immediate feed- Table 1). ing response by attempting to attack. Based on these ¿ ndings, we can con- Rodriguez-Robles and De Jesus-Es- clude that snakes kept for generations in cobar (1999) studied evo- captivity behave less actively than those lutional relationships by analyzing the ¿ eld collected or those with the addition mt DNA variability and adjusting their of “wild blood”. We may ask whether conclusions to the evolution of feed- decreased activity reÀ exes lower ¿ t- ing customs. According to the authors, ness ability. Each group of reptiles has a diet based on reptiles (lizards, snakes) its own survival strategy. Research car- is more primitive than one based on ro- ried on turtles (Mrosovsky and Gogfrey dents, which developed much later. They 1995) has shown that the more active presented their results in the form of young (1YO) specimens have a lower dendrograms, showing both the genetic survival rate than those less mobile, relations between genera of American whereas the higher mobility of young colubrids and the hypothetic evolution lizards (Van Damme et al. 1992) pro- of feeding behaviour. vides them a greater success rate in hunt- The results presented earlier corre- ing and a greater ability to avoid preda- spond with those quoted. Divergence tors. If greater activity and courage in in behaviour during confrontations with young snakes protects them in the wild, predators philogenetically approximates researchers may conclude that a longer feeding behaviour and differs by de¿ ni- generation stage in captivity debilitates tion from inner aggression. adaptation and ¿ tness. This is true for the The divergences noted above are very ¿ rst stages of life only, as during the II important from the point of view de¿ ned observation all groups behaved similarly in the title of this study. During the ¿ rst 250 A. ĩyczyĔski, Z. Nowak trials, we observed diversity in both the captivity as a function of potential, unde- ¿ tness of the specimens and their reac- sirable selection. tions during confrontation. These dif- Snakes play an important role in the ferences were reduced as the snakes got trophic chain. In the event of reintroduc- older. So any decision to reintroduce tion of captive material into the natural snakes should only consider very young habitat, the consequences of releasing specimens, i.e. hatchlings. Such mate- animals of genetically changed feeding rial may be characterized by greater ¿ t- customs could be at the least undesirable, ness – vide Table 1, although due to their even where the rules of taxonomic and smaller size, these could be more at risk population ¿ delity have been obeyed. It from predators. But this would allow for is worth mentioning that the team work- natural selection, a factor always present ing on the reintroduction of the Smooth in the natural habitat. Snake Coronella austriaca (ZieliĔski and The accordance of reactions with the Stanisáawski 2001) were offering their dendrograms shown by Rodriguez-Rob- captive hatched snakes live small lizards les and De Jesus-Escobar revealed that (Lacerta agilis), which were preyed on. captive Lampropeltis snakes preserved This reÀ ected in terrarium conditions the their natural feeding behaviour. Thanks rules of natural selection for the snakes’ to their undisputed beauty, Kingsnakes survival. However, the results of this and Milk Snakes have gained enormous restocking are dif¿ cult to evaluate, as popularity among amateur snake keep- the snakes were not tagged for telemet- ers. In captive conditions, they are forced ric monitoring as they were in Daltry’s to adapt to a based diet. Some (1999) project. specimens adapt to this easily; others Lack of proper documentation is al- are more reluctant. Nevertheless their ways the weakest point of any reintro- maintenance in captivity is increasingly duction project, as was pointed out by easy and the survival rate of hatchlings is Earnhardt (1999). This particularly con- gradually increasing. This fact is worthy cerns survival rates in the very ¿ rst pe- of attention. The most popular terrarium riod of release. species may have been subjected to un- The dendrograms and observations intentional selection for the ability to presented in this paper lead to conclution adapt to a diet that is atypical for them. that studied Lampropeltis snakes, despite Arnold (1981), working on garter snakes their history of captivity, preserved their Thamnophis elegans proved that feeding natural feeding habits, enabling them to preferences are genetically polymorphic play their natural role in the trophic chain and of a quantitative nature. Thus selec- when returned to their natural environ- tion for diet type could be possible and ment. Likewise the Pantherophis snakes, may also result in inherited changes in despite their level of domestication, did feeding customs, not caused by current not differ from one another in the general arti¿ cial captive conditions only. This set of reactions. problem is discussed by Conway 2011, focusing on the number of generations in Comparative behaviour analysis of colubrids with... 251

CONCLUSIONS RODRIGUEZ-ROBLES J.A., DE JESUS- -ESCOBAR J.M., 1999: Molecular sys- This model experiment shows that cap- tematics of New World Lampropeltinine tive snakes from multigeneration terra- Snakes (Colubridae): implications for rium colonies can be considered worthy biogeography and evolution habits. Bio- material for reintroduction, on condition logical Journal of the Linnean Society 68: 355–385. that the principles of genetic ¿ delity are MARKOWSKA A., 1979: Reakcja na zmianĊ observed. bodĨca wzrokowego u szczurów z uszko- A planned release should be carried dzeniem kory przedczoáowej, prąĪkowia out at a very young age, despite the ex- lub formacji hipokampa. Praca doktor- pected losses of very small and fragile ska. Instytut im. M. Nenckiego PAN. animals, in order to let them undergo MATTISON CH., 1995: The encyclopedia a process of natural selection based on of snakes. Blandford Press, New York. MOSEBY K.E., READ J.L., PATON D.C., fully exposed diversity. COPLEY P., HILL B.M., CRISP H.A., 2011: Predation determines the outcome of ten reintroduction attempts in arid REFERENCES South Australia. Biological Conservation ALEXANDER G., MARAIS J., 2007: A 144 (12): 2863–2872. guide to Reptiles of Southern Africa. MROSOVSKY N., GODFREY M.H., 1995: Struik Publishers, Cape Town. Manipulating sex ratios: turtle speed ARNOLD S.J., 1981: Behavioural variation ahead! Chelonian Conservation and Bi- in natural populations II. The inheritance ology 1 (3): 238–240. of a feeding response in crosses between NEI M., 1972: Genetic distance between geographic races of the garter snakes populations. American Naturalist 106: Thamnophis elegans. Evolution 35(3): 283–291. 510–515. SPANOWICZ M., ĩYCZYēSKI A., 2002: CONWAY W.G., 2011: Buying time for wild Projekt zasilania lokalnej populacji Lam- animals with zoos. Zoo Biology 30: 1–8. propeltis alterna w rejonie Limpia Can- DALTRY J., 1999: Interim report – Re-intro- yon (Pád.-Wsch. Teksas). Przegląd Przy- duction of the Antiguan Racer (Alsophis rodniczy 13 (3): 147–150. antiguagae). Antiguan Racer Conserva- STANISàAWSKI W., 2003: Hodowla kra- tion Project. www.eco-index.org/search/ jowych gadów w polskich ogrodach pdfs/1418report_6.pdf. zoologicznych. Materiaáy z I Spotkania DALTRY J., 2006: Reintroduction of the Herpetologów Polskich Ogrodów Zoolo- critically endangered Antiguan Racer Al- gicznych. àódĨ 27–28 listopada 2003. sophis antiguagae to Rabbit Island Anti- Van DAMME R., BAUWENS D., BRA- gua. Conservation Evidence 3: 33–35. NA F., VERHEYEN R., 1992: Incubation EARNHARDT J.M., 1999: Reintroduction temperature differently affects hatching programmes: genetic trade-offs for the pop- time, egg survival and hatchling perfor- ulations. Animal Conservation 2: 279–286. mance in the lizard Podarcis muralis. GODWIN J., JOHNSON V., GUYER C., Herpetologica 48 (2): 220–226. RUSH M., 2008: Captive propagation ZIELIēSKI P., STANISàAWSKI W., 2001: of the Eastern Indigo Snake for reintro- Ochrona gniewosza plamistego w Polsce duction into Alabama. Report. Alabama ĝrodkowej – wyniki wstĊpne. Materiaáy Dept. of Conservation and Natural Re- z konferencji „Metody czynnej ochrony sources Montgomery. Unpublished. herpetofauny”. àagów Lubuski 17–18 listopada 2001. 252 A. ĩyczyĔski, Z. Nowak

Streszczenie: Porównawcza analiza zachowaĔ naturalnej diecie, wystąpiáy normalne reakcje wĊĪy z rodziny Colubridae pod kątem przydat- drapieĪnicze. Stwierdzono zatem przydatnoĞü noĞci osobników hodowanych w niewoli do rein- populacji terraryjnej jako ewentualnego banku trodukcji do Ğrodowiska naturalnego. W modelo- genów dla strategii ochronnej ex situ, przy zalece- wym doĞwiadczeniu przetestowano amerykaĔskie niu uĪywania do reintrodukcji materiaáu moĪliwie wĊĪe z rodziny Colubridae podzielone wedáug máodego, najwierniej reagującego na presjĊ se- stopnia udomowienia (pokoleniowego staĪu lekcji naturalnej, wymierzonej w wachlarz reakcji w warunkach niewoli). ZwierzĊta zostaáy podda- niestáumionych warunkami niewoli. ne testowi otwartego pola (OFT), sprawdzającego ich aktywnoĞü i odwagĊ na otwartej przestrzeni MS. received in November 2013 oraz konfrontacji z innym wĊĪem zboĪowym Pantherophis guttatus oraz lancetogáowem kró- Authors’ address: lewskim Lampropeltis getula splendida i dodat- Andrzej ĩyczyĔski, Zuzanna Nowak kowo L.g. californiae, formą albinotyczną. Test Szkoáa Gáówna Gospodarstwa Wiejskiego OFT wykazaá ujemny wpáyw wieku na aktywnoĞü w Warszawie terraryjnych zwierząt a konfrontacje z probanta- Wydziaá Nauk o ZwierzĊtach mi nie ujawniáy wpáywu udomowienia na reak- Katedra Genetyki i Ogólnej Hodowli Zwierząt cje obronne (klasterowa analiza skupieĔ). Nawet ul. Ciszewskiego 8, 02-786 Warszawa u albinotycznych lancetogáowów, uznanych za Poland najbardziej udomowioną formĊ (zuboĪaáy, wsob- e-mail: [email protected] ny genotyp) utrzymywanych od pokoleĔ na nie- [email protected]