Ex situ Management of

Brij Kishor Gupta | Benjamin Tapley Karthikeyan Vasudevan | Matt Goetz

ASSAM STATE ZOO CUM BOTANICAL GARDEN Ex situ Management of Amphibians

Brij Kishor Gupta Benjamin Tapley Karthikeyan Vasudevan Ma Goetz

Price INR : 300.00

Published by: Assam State Zoo cum Botanical Garden, Guwaha, Assam,

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First Edion

Copyright : © 2015 Brij Kishor Gupta

Informaon on this tle : [email protected] [email protected]

Cover Photo Credit :

1. Indian baloon globulosum: Navendu Page 2. Spade footed burrowing frog brevicepts: Navendu Page 3. Venated cliding frog pseudomalabaricus: S. U. Saravanakumar 4. Himalayan Newt Tylotoriton verrucosus: Karthikeyan Vasudevan

Citaon: Gupta, B.K., Tapley, B., Vasudevan, K., and Goetz, M., (2015). Ex situ Management of Amphibians. Assam State Zoo cum Botanical Garden, Guwaha, Pp. 72

Precauonary Note:

Amphibian husbandry is a rapidly developing field and it has become apparent that amphibians have complex and varied husbandry requirements and are not necessarily easy to maintain and breed. Many have not been maintained in capvity, therefore developing appropriate and opmal husbandry and breeding techniques is crucial to the success of ex situ programmes. Tropical species in parcular have proven difficult to establish in capvity, due to high levels of endemism combined with a tendency towards high levels of environmental specificity and limited husbandry experience or field data. Herein we present a number of species specific husbandry guidelines. Whilst some of these guidelines are based on direct experience either with the specific species or a closely related species, others (E.g. , , Ramanella and Uperodon) have been compiled based on how we would go about maintaining a species in capvity based on field experience and literature reviews. Ex situ Management of Amphibians

Brij Kishor Gupta | Benjamin Tapley Karthikeyan Vasudevan | Matt Goetz

ASSAM STATE ZOO CUM BOTANICAL GARDEN Acknowledgements

Authors of this book are thankful to friends and colleagues for their help in bringing out this publicaon.

We thank Mr. Bishan Singh Bonal, Member Secretary, CZA for suggesng to bring out present publicaon.

We thank many people at Durrell Wildlife Conservaon Trust, Jersey, Channel Islands – Dr. Lee Durrell, Honorary Director, Dr. Tim Wright, Academy Manager, Dr. Andrew Routh, Head of Veterinary Services, and Mr. Ma Goetz, Head of Herpetology – for providing guest lectures and technical inputs during the workshop and inputs to this publicaon.

We thank Director, Zoological Society of London for allowing Mr. Benjamin Tapley to deliver guest lecture during the workshop and contribute for this book.

We thank Director, Wildlife Instute of India, Dehradun for technical inputs.

We thank Director, CCMB/Laboratory for Conservaon of Endangered Species, Hyderabad for their technical support and allowing Dr. Karthikeyan Vasudevan to be part of this publicaon.

We thank Mr. Chandan K. Bora and his staff at Assam State Zoo & Botanical Garden, Guwaha for hosng the workshop.

We thank Chief Wildlife Warden, Government of Assam and Forest Department of Assam as a host of the workshop.

Last but not the least thanks to fellow zoo directors, curators, biologists who aended the workshop and their valuable contribuon in developing this publicaon.

ii Foreword

Lee Durrell Honorary Director, Durrell Wildlife Conservaon Trust

The global crisis – the unprecedentedly sudden and rapid decline of a whole class of vertebrate worldwide – is now generally acknowledged, as is the key role that ex situ conservaon programmes will play in addressing it.

Of all species of globally threatened amphibians, however, zoos hold only 6.2%, far short of the number for which ex situ management is considered necessary. Over the last 20 years zoos have increased their efforts to aid amphibian conservaon, but more aenon must now be turned to species which have been named as high priories for coordinated conservaon breeding programmes.1

With a rich amphibian fauna to which newly discovered species are frequently added, India also harbours several internaonally recognised hotspots for amphibian diversity. Its zoos have a vital role to play in safeguarding this important aspect of its natural heritage, as well as seng an example for the rest of the world's zoo community to follow.

The iniaves taken since 2008 by the Central Zoo Authority to promote the role of zoos in the conservaon of Indian amphibians culminated in a workshop late in 2013 in Guwaha. There was excellent representaon from Indian zoos, and invited experts came to discuss with their colleagues the way forward for amphibian conservaon in India.

Quite rightly, the workshop focussed on first things first – the priorisaon of species in need of ex situ conservaon and the development of experse and capacity in husbandry and breeding by using model species.

This book gleans the knowledge, experience and skills of many people who have managed a wide range of amphibian taxa in capvity. These can be applied to model or 'pracce' species in order to hone the competences needed for coordinated conservaon breeding programmes for targeted threatened species. Illustrave of the pragmac approach is the naming of 23 zoological instuons in India and the assignment of 'pracce' species to them.

Let us hope that what is learned through 'pracce' is put to the test sooner rather than later by the parcipang zoos, not only for the sake of India's threatened amphibians, but also to enrich amphibian conservaon planning at a global level.

1Dawson, J., Patel, F., Griffiths, R.A. and Young, R.P. 2015. Assessing the global zoo response to the amphibian crisis through 20 year trends in capve collecons. Conservaon Biology DOI: 10.1111/cobi.12563 Dated : 5 OCTOBER 2015 iii

Preface

Smithsonian Conservation Biology Institute

This booklet, Ex‐situ Management of Amphibians is needed now, more than ever to keep populaons afloat and save species from exncon. Amphibians are declining rapidly in many parts of the world, in some cases, before even being officially known to science. The amphibian crisis has led to the creaon of a Global Amphibian Ark organizaon, coordinated by the World Associaon of Zoos and Aquariums (WAZA), the Conservaon Breeding Specialist Group (CBSG) of the IUCN's Species Survival Commission, and the Amphibian Specialist Group (ASG). The goal has been to mobilize a truly global network of actors, including building serious capacity to establish genecally viable ex situ populaons of the most endangered amphibians. These intervenons are benefing amphibians by buying us me to figure out how to migate the threats causing the precipitous declines in nature.

One interesng component to ex situ management and breeding of amphibians is that so much informaon has remained 'stuck' in people's heads rather than being wrien down. There also are huge gaps in knowledge. Amphibians have evolved a wide range of life‐history traits that help them survive and thrive in habitats ranging from dry deserts to tropical forests to rugged mountaintops. This diversity means that keeping them successfully in capvity oen can be considered an art form, pracced best by dedicated students of natural history and zoology. Many discoveries await the careful, aenve observer who cares for these species, documenng (and wring down) their success and failures through trial and error. This strategy of adapve management accompanied by a scienfic approach to answer key quesons in nutrion, reproducon and welfare (among other disciplines) is desperately needed now. Done well, we can improve our knowledge of amphibian husbandry to a point where we have a decent chance of establishing healthy ex situ assurance colonies that, as possible, can be used later for reintroducons to the wild.

But first things first. In reality, we know lile about the basic natural history of many of India's 384 amphibian species, and even less about how to breed them. But the guidelines provided here provide a welcome and crical first step, especially as the informaon shared is based on real life experiences by experts with vast, praccal experiences on a diversity of taxa. We hope others will be spurred on to embark on ark projects of their own, remembering to study not only endangered species, but also the most common. Aer all, the study of non‐threatened species (as surrogates) oen provides us the best strategies for saving the rarest. Regardless, we predict that the observaons and recommendaons provided in this booklet will inspire others to increase both knowledge and ex situ conservaon breeding of this incredibly diverse class of and .

Brian Gratwicke Amphibian Conservaon Biologist Smithsonian Conservaon Biology Instute

David Wildt Head Center for Species Survival Smithsonian Conservaon Biology Instute

Dated: 22nd September, 2015

v

Contents

Acknowledgements ii

Foreword iii

Preface v

1. Introducon 1

2. Ex‐situ Management of Terrestrial species 4

3. Ex‐situ Management of Microhylid frogs , , Ramanella & Uperodon species 11

4. Ex‐situ Management of Leaping frogs Indirana species 18

5. Ex‐situ Management of Bicoloured frog Clinotarsus curpes 24

6. Ex‐situ Management of Common tree frogs species 31

7. Ex‐situ Management of Gliding frog Rhacophorus species 38

8. Ex‐situ Management of Salamander Tylototriton verrucosus 45

9. Marking of Amphibians 51

10. Culturing Food for Amphibians 53

11. Enrichment of Amphibian Exhibits 56

12. Amphibian Diseases 64

13. Priorized Species : List of target species and pracce species with the name of zoological parks idenfied for iniang the amphibian housing, exhibit and if required, planned conservaon breeding in India 69

Ex situ Management of Amphibians 1. Introduction threats that they face. Currently amphibians are underrepresen- W o r l d w i d e , a m p h i b i a n ted in Indian Zoos and only one populations are declining1, and s p e c i e s ( i . e . S a l a m a n d e r India has about 390 species and T y l o t o t r i t o n v e r r u c o s u s ) more than 70% of them are maintained by one institution, endemic to the region. It also Padmaja NaiduHimalayan harbours more threatened Zoological Park in Darjeeling. species of amphibian than any The Central Zoo Authority other country in the Indo recognises the need to increase Malayan realm2 The Western capacity in amphibian zoos. Ghats and Eastern Himalaya are r e c o g n i s e d b i o d i v e r s i t y The Central Zoo Authority (CZA) hotspots, with high levels of has made several attempts to amphibian endemism3,4,7. The s t r e n g t h e n c o n s e r v a t i o n amphibians of the Western b r e e d i n g p r o g r a m m e f o r Ghats are enigmatic and new amphibians, by providing species are being described with training to zoo professionals and astonishing frequency, it is promoting ex-situ conservation estimated that just 50% of efforts in different Indian Zoos. amphibians in the South Asian In February 2008, in the region have been described5 with I n t e r n a t i o n a l C o n f e r e n c e new amphibian genera being organized by CZA on India's described as recently as 20136. Conservation Initiative the c o n s e r v a t i o n b r e e d i n g programme of Himalayan Salamander was reviewed. In January 2009, workshop on ex- situ conservation for amphibians was organized by CZA in Mysore where preliminary attempts w e r e m a d e t o p r i o r i t i z e amphibians for conservation breeding in Indian zoos. In D e c e m b e r 2 0 1 3 , t h e C Z A organised a workshop “Building Figure 1 Bronze frog (Hylarana temporalis) (Brij Kishor Gupta) National Capacity for ex-situ Indian Zoos could play a pivotal Amphibians Management and r o l e i n t h e c o n s e r v a t i o n Conservation” in Guwahati, management of the countries where a list target and practice threatened amphibians. In species of amphibians were addition zoos are ideally placed identified. During this workshop to educate the visiting the public the Central Zoo Authority with about amphibians and the the assistance of the Durrell 1 Ex situ Management of Amphibians Wildlife Conservation Trust and the Zoological Society of London has strengthened national c a p a c i t y i n a m p h i b i a n management. More than 80 delegates from all over India r e p r e s e n t i n g n e a r l y 4 0 institutions participated in these workshops. The participants were exposed to the specific requirements of amphibians in the design and management of ex-situ facilities. Participants developed hands-on skills in e n c l o s u r e d e s i g n , t h e management of water flow and Figure 3 Wrinkled frog (Nyctibatrchus species) (Brij Kishor Gupta) quality, temperature and light Over the years CZA has paved within the captive facility and the way for prioritisation of species p r i n c i p l e s o f a m p h i b i a n and preparation of a plan for n u t r i t i o n , r e p r o d u c t i o n , coordinated conservation biosecurity and conservation breeding for Indian amphibians. e d u c a t i o n . T h e p o t e n t i a l With ecology and biology of c o n t r i b u t i o n t h a t e x s i t u many amphibians in India amphibian management could r e m a i n s u n k n o w n , i t i s provide to in situ amphibian potentially difficult to keep, conservation was also covered. establish and breed Indian amphibians. We strongly urge y o u n g b i o l o g i s t s a n d z o o p r o f e s s i o n a l s t o g a t h e r information through targeted studies on species in the field before embarking on captive breeding programmes. A step in that direction would be to indentify a model amphibian species for the zoo to practice husbandry techniques with, nevertheless a vital first step for p r o m o t i n g a m p h i b i a n conservation breeding in Indian zoos. The present guidelines on Figure 2 Narrow mouthed frog (Microhyla rubra) (Brij Kishor Gupta) the ex situ management of amphibians are part of output of

2 Ex situ Management of Amphibians the workshop on “Building G u n a t i l l e k e , P . V . National Capacity for ex-situ Karunakaran, K.G. Nayak, S. Amphibians Management and Prasad, P. Puyravaud, B.R. Conservation” held at the Assam R a m e s h , K . A . State Zoo, Guwhati, Assam, Bubramanian & G. Vasanthy India during December, 2013. .2007. A brief overview of the – biodiversity hotspot.| References C u r r e n t S c i e n c e . 1 1 : 1567–1572 1. R.A. Alford, S.J. Richards. 1999. Global amphibian 5. S. Molur. 2008. South Asian declines: a problem in amphibians: , applied ecology. Annual diversity and conservation Reviews in Ecology and status. International Zoo Systematics. 30: 133–165. Yearbook. 42: 143–157.

2. S.N. Stuart, M. Hoffmann, 6. R.K. Abraham, R. Alexander J.S. Chanson, N.A. Cox, R. P y r o n , B . R . A n s i l , A , Berridge, P. Ramani, B.E. Zachariah, A. Zachariah. Young, Eds., Threatened 2013. Two novel genera and Amphibians of the World. one new species of treefrog (Lynx Ediciones, with The (Anura: ) World Conservation Union, highlight cryptic diversity in Conservation International, the Western Ghats of India. a n d N a t u r e S e r v e , Zootaxa 3640: 177-199. Barcelona, 2005) 7. Gascon, G., Collins, J. P., 3. N. Myers, R.A. Mittermeier, Moore, R. D., Church, D. R., C.G. Mittermeier, G.A.B. da M c K a y , J . E . a n d Fonseca, J. Kent. 2000. Mendelson, J. R. III (eds). Biodiversity hotspots for 2 0 0 7 A m p h i b i a n conservation priorities. Conservation Action Plan. Nature 403: 853–858. I U C N / S S C A m p h i b i a n Specialist Group. Gland, 4. N.R. Gunawardene, A.E.D. Switzerland and Cambridge, D a n i e l s , I . A . U . N . UK. 64 pp. G u n a t i l l e k e , C . V . S .

3 Ex situ Management of Amphibians 2. Ex-situ Management of Terrestrial toads Bufo species

and taxonomic revision of the complex is required (IUCN et al., 2006).

Habitat and Ecology melanostictus are very adaptable and occurs in a wide variety of habitats, including beach vegetation, various human habitats, natural edge habitats, and closed subtropical or tropical forest (IUCN et al., 2006). Figure 1. Duttaphrynus melanostictus (Benjamin Tapley). Sexing Individuals Introduction Duttaphrynus melanostictus are sexually dimorphic with females being Description more than 50% larger than males. Terrestrial toads are stocky bodied, Reproductively active males have short limbed and possess parotid glands nuptial pads on the inside of the 1st and b e h i n d e a c h e y e ( F i g . 1 ) D . 2nd fingers (Fig. 2). Daniels, 2005 m e l a n o s t i c t u s i s p o l y m o r p h i c , reports that in some parts of their range colouration and patterning are highly males turn yellow during the breeding variable, even amongst wild specimens season (Fig. 3). collected in the same areas. This species can be characterised by the black tipped bony warts distributed all over its body and hardened ridges on its head ( D a n i e l s , 2 0 0 5 ) F e m a l e s o f D . melanostictus may grow to 150 mm snout vent length (SVL) and males up to 54 mm SVL (Li Fan et al, 2013).

Distribution Duttaphrynus melanostictus is a widely distributed species, occurring from Pakistan to southern and south throughout southern and south east Asia and parts of Indonesia. It can be found from sea level up to 1, 800 m asl (IUCN et al., 2006).

Conservation Status and Threats Duttaphrynus melanostictus is listed as Least Concern by the IUCN Red List due to its extensive distribution and its tolerance of a wide range of habitat types(IUCN et al., 2006).. It should be Figure 2. Top. Typical black nuptial pads seen on the first noted, however, D. melanostictus and secnd finger of terrestrial toads (Benjamin Tapley). Figure 3. Male D. melanostictus exhibiting yellow breeding represents a complex of cryptic species, colours (Benjamin Tapley) 4 Ex situ Management of Amphibians

Reproduction and Larval implant elastomer (VIE) but this may Development not be necessary for rearing purposes I n d r i e r p a r t s o f i t s r a n g e , D . melanostictus, breeds during the Housing adults monsoons (Amphibiaweb). In wetter Duttaphrynus melanostictus require areas of its range (Including north east relatively large enclosures a floor space India the species reportedly breeds year of 600 x 100 mm would be the minimum round (Church, 1960; Ahmed, 2009). enclosure size for 5 individuals (Fig.4). Males congregate around slow moving Height is not important as these toads or still water bodies where they vocalise are poor climbers but the height of the (Daniels, 2005). enclosure should allow them to hop freely without coming into contact with Amplexus, Egg Laying and Larval the lid, an enclosure height of 500 mm development would be sufficient. A wire mesh lid T h e b r e e d i n g b i o l o g y o f D . should be provided. A 250 x 150 x 100 melanostictus is well documented. mm water dish should be provided and Amplexus (mating position) is axillary in should be deep enough for the toads to this species. Once in amplexus the immerse their drink patches. female deposits up to 4,000 eggs (Li Fan et al, 2013) which hatch after 48hours Cover objects are essential when ( D a n i e l s , 2 0 0 5 ) . I n C h e n n a i keeping terrestrial toads. 150 mm metamorphoses takes approximately 45 lengths of PVC pipe or hollowed bamboo days (Daniels, 2005). make appropriate cover objects, these need to be able to accommodate the Longevity and Age at Sexual toads so the size of the tube will depend Maturity on the size of the . Live plants can be Lifespan in the wild is unknown. In added to the enclosure as they create captivity D.melanostictus have been humid pockets which the may recorded to live for over ten years use, the plants also provide visual (Csurhes, 2010). Sexual maturity is at barriers for the toads. least partially governed by size, they are reported to reach sexual maturity when Lids should be constructed of mesh as they are 23.0 g (Csurhes, 2010). glass will block UVB.

Captive Management Substrate must be provided for this species as they like to dig. The provision of substrate will prevent the enclosure Introduction from drying out. Be sure to incorporate a Duttaphrynus melanostictus is drain in the tank construction. A layer of commonly kept by zoos and private gravel covered with a permeable matting individuals, and have been bred on should be laid underneath a substrate of occasion in captivity by private breeders. soil mixed with sand (ratio 2:1). The depth of substrate will depend on Identifying Individuals enclosure size and the size of the Adult specimens of both species are not occupants, but must be at least 100 mm individually identifiable as specimens deep. The larger the enclosure, the do not possess characteristic markings. deeper the substrate should be as the Adults could be marked with passive substrate will dry out quickly in large integrated transponders (PIT tags). Pit enclosures. Substrate does not need to tags should be inserted subcutaneously be compacted for this species. Ensure on the dorsum. Juveniles could be that substrates are not contaminated marked with different coloured visible with fertilisers or other substances that

5 Ex situ Management of Amphibians may be toxic to amphibians. lighting is inaccessible to the frogs as Ensure the substrate has a humidity they will be burnt easily if they come into gradient by pouting some water (or contact with lamps. spraying with water) more at the end of the enclosure with no basking light. Check the source of your water by testing it regularly for ammonia, nitrite, nitrate, pH and phosphates. Tap water should be left to stand for 24 hours before it is used in order for the chlorine to dissipate. Ensure the temperature of the water is at equilibrium with the room temperature. A plastic water dish should be provided and should measure 200 x 150 mm. Water depth should be 80 mm, and toads should be able to immerse their drink patch.

Figure 4. Set up suitable for housing and breeding An emergent rock should be placed in D. melanostictus (Zoological Society of London) the water to allow the frogs easy access Temperature, Humidity and out of the water. Lighting Husbandry should be informed by data The enclosure should be lightly misted collected from the field. Basking spot with aged tap water daily. temperatures, UV exposure and gradients, temperature and humidity Routine Husbandry r e g i m e s s h o u l d m i m i c t h e All animals should be visually inspected environmental conditions at the daily. Water dishes must be cleaned out collection site. Note that environmental and refilled daily as amphibians often data should be collected at different excrete waste products in water. Water times throughout the day and year. quality cannot be judged by eye. Faeces Suitable ambient room temperatures and uneaten food must be removed should vary between 23 and 27˚C daily. The enclosure and all furniture (night/day summer) and 20 and 25˚C must be thoroughly scrubbed (with a (night/day winter). brush and hot water, no chemical cleaners or disinfectants) twice a week. A UV gradient should be provided in Temperatures of the enclosure should be conjunction with a basking spot. Arcadia checked at different points of the D3+ reptile lamp T5 12% UVB are ideal enclosure regularly to ensure that the and a maximum UVI of 3 should be correct thermoregulatory gradients are provided for the frogs at their basking maintained. UV readings must be taken spots. Remember that mesh will filter every three months to check that there is out some of the UVB so UV readings a sufficient UV gradient and that the UV must be taken from below the mesh with emitting lights are still emitting UV. a solarmeter 6.5. The substrate must be changed as soon Basking spots can be provided with as it looks dirty or if it becomes saturated regular incandescent lamps. The or unable to retain humidity. basking lamp should be positioned at one end of the enclosure and the Diet in captivity temperature on the substrate below the Most toads tend to favour small food basking spot should range between 30- o o items and D. melanostictus have been 35 C and not exceed 35 C. Ensure that all observed feeding on and winged 6 Ex situ Management of Amphibians in India (Daniels, 2005). Great into the rain chamber to maintain water care should be taken when feeding quality. Ideally a sump could be used captive amphibians ants as ants can be aggressive and may bite and sting the toads, this is problematic because the toads cannot escape the ants if they choose to. Daniels, 2005 reports that D. melanostictus avoid armoured prey items such as beetles and crickets with hard exoskeletons. Some of the commercially available crickets are f a i r l y s o f t b o d i e d a n d m a y b e appropriate. Food should be offered after it is dusted with an appropriate dietary supplement, well fed and an appropriate size, no greater in size than t h e w i d t h o f t h e f r o g s h e a d . Duttaphrynus melanostictus are nocturnal and should be fed in the evening.

Adults should be fed every three to six Figure 5. A rain chamber suitable for breeding days (depending on season and D. melanostictus. (Benjamin Tapley) condition), juveniles up to six weeks of (Appendix 1). The, lighting and age should be fed daily. temperature parameters should be maintained as above. Visual barriers Reproduction in Captivity such as palm fronds or plastic plants could be used to provide cover for the Breeding Enclosure toads in their breeding enclosure if they fail to spawn after a couple of weeks. Increasing the humidity can trigger breeding in D.melanostictus. These toads should be cycled in a rain chamber The toads should be fed at least every with a sprinkler system for breeding other day whilst they are in the rain (Fig. 5). chamber but ensure that any dead insects are removed daily as they will pollute the water. A rain chamber with a floor space of 600 x 1000 mm would be suitable as a minimum size for 2 males and 3 females. In closed systems regular water tests It is important that the sex ratio in the should be carried out (Temperature, pH, breeding enclosure is not biased towards ammonia, nitrite and nitrate), the males as toads often form mating balls results will help you decide the which may result in the drowning of the frequency of water changes. pH should females (Mathew & Sen, 2010, ). No not fluctuate, ammonia should measure substrate is required in the rain less than 1 part per million (ppm), nitrite c h a m b e r . M o s t t o a d s a r e p o o r less than 0.1ppm and nitrate less than swimmers so it is vital that there are 10ppm. No more than 20% of the water some rocks or logs in the water that the should be changed as rapid shifts in toads can easily leave the water should water quality will be problematic for the they choose to do so. A pump powered biological filtration process. spray system or fountain would be ideal for breeding this species but a mature Breeding Seasonality biological filter should be incorporated There is no published report on the

7 Ex situ Management of Amphibians captive breeding of D. melanostictus. The size of the tank is not important, the stocking density is. Tanks should be stocked with no more than 5 tadpoles Provision of Breeding Sites per litre. Filtration is vital (see appendix A depth gradient should be provided for one to inform your choice of filter). Aged the toads so that they can choose their tap water can be used to rear tadpoles. egg laying site, this can be provided by Check the source of your water by testing angling slates or paving slabs into the it regularly for ammonia, nitrite, nitrate, water (Fig.6). hardness, pH and phosphates. Water parameters should be informed by field data but remember that not all breeding sites may have optimal water quality, especially in urbanised environments where these toads can often be found. Tap water should be left to stand for 24 hours before it is used in order for the chlorine to dissipate. Ensure the t e m p e r a t u r e o f t h e w a t e r i s a t equilibrium with the room temperature. Check the water in closed systems and c a r r y o u t r e g u l a r w a t e r t e s t s (temperature, pH, ammonia, nitrite and nitrate), the results will help you decide the frequency of water changes. pH Figure 6. A basic rain chamber used to successfully breed should not fluctuate, ammonia should the Vietnamese bony headed toad (Ingerophrynus galeatus) in captivity. Note the sloped tile which provided the toads measure less than 1 part per million with a depth gradient in the rain chamber (Benjamin Tapley). (ppm), nitrite less than 0.1ppm and nitrate less than 10ppm. No more than Egg Care, Tadpole Husbandry & 20% of the water should be changed as Development rapid shifts in water quality will be Most toads lay a paired string of eggs. It problematic for the biological filtration is important that the eggs (Fig. 7) are and tadpoles. Ideally a sump should be transferred to a mature aquatic system used to increase the volume of water of soon after laying so that they are not your system and to house all the life damaged or exposed to the air by the support systems (filters etc). If using an toads moving around in the rain internal canister filter ensure that it is chamber. You may not wish to keep all not too strong as the tadpoles could be the eggs as rearing up to 4,000 tadpoles sucked into the inflow of the filter and will be a huge task. may have difficulty feeding. Direct the

Figure 7. Paired egg string typical of toads. Figure 8. A typical toad tadpole is black in colour 8 Ex situ Management of Amphibians outflow to the surface of the water for oxygenation.

Toad tadpoles are typically black in colour. When they first hatch they will not move and it is perfectly normal for them to sometimes rest on their side. If you look on the underside of tadpoles when they first hatch you should be able Figure 9. Enclosure for rearing juvenile terrestrial toads to see the yolk sac. The tadpole will not (Benjamin Tapley). start swimming or feeding until this ac has been absorbed. Once the newly Rearing Metamorphs hatched tadpoles have dropped off the Metamorphs can be housed in small egg strings it is important to remove the plastic containers. A 350 x 200 x 300 remaining egg jelly as this will decay and mm enclosure is suitable for up to 12 pollute the water. Substrate is not individuals, these groups should be necessary but tadpoles like to hide. divided amongst larger enclosures as D r i e d I n d i a n a l m o n d l e a v e s individuals grow. It is important that (Terminalia catappa) can be added, rearing tanks are relatively small so that they also soften the water which would high ambient humidity can me be beneficial in hard water areas. Partial maintained and that the juvenile toads (10-20 %) water changes should be can find food easily. Covering the lid carried out regularly, the frequency of with a fine breathable cloth or fleece will water changes must be governed by prevent the food escaping and ensure frequent water testing. that humidity remains high. The cloth will block dome of the UV emitted from Tadpoles should be fed daily, once they the lights so you must experiment and start free swimming. They should be fed e n s u r e t h a t t h e U V l i g h t a n d on a variety of food. At Durrell the temperature gradients are still suitable. tadpoles of the Vietnamese bony headed Compacted damp soil mixed with sand toad were successfully reared on a would make a good substrate, and this powdered tadpole food (components: must be replaced as necessary. It is ground tropical fish flake, grass pellet, important that the substrate is trout pellets, tubifex, river shrimp, compacted when housing metamorphs spirulina algae and cuttlefish bone). as the soil particles easily stick to the Food should be available to the tadpoles skin of the toadlets. A shallow water dish at all times but it is important that large must be provided (water depth 10 mm). amounts of uneaten food do not pollute It is essential that the water dish is easy the aquarium. Tadpole feeding regimes for the metamorphs to climb out of (a must be modified, as the tadpoles grow rock or small branch in the water dish they will eat more. facilitates access). When transferring toadlets from the tadpole tank make sure that they are placed in the water Once the front limbs appear and the tail dish and not on the substrate as they starts to reduce it is important that the may not be quite ready to leave the metamorphosing toadlets are given the water, they will choose when they are opportunity to emerge from the water as ready. Cover objects are extremely they can easily drown. Emerging rocks important microhabitats for small or aquatic plants are suitable. Once they toads. Bark hides and leaf litter should emerge they are prone to drying out and be added to the enclosure. The easily die if the humidity is not high. furnishings in th enclosure should not be too complex or it will be difficult for the y o u n g t o a d s t o fi d f o o d . T h e

9 Ex situ Management of Amphibians metamorphs can be raised at the same Bibliography room temperature as the adults (23 - 27˚C), and must be misted with room Ahmed, F.M., Das, A & Dutta, S.K temperature tap water twice daily. It is vital that metamorphs and juveniles are (2009) Amphibians and reptiles of provided with UV at this critical growing North East India. Aaranyak, stage (Arcadia D3+ reptile lamp T5 12% Guwahati. 168pp UVB), this must be given in a gradient with a maximum UVI of 2 (Fig. 9). Csurhes, S. (2010) Pest risk assessment; Asian spined toad o Do not provide metamorphs with a 35 C (Bufo melanostictus). Department t basking spot as this will dry them out o f E m p l o y m e n t , E c o n o m i c rapidly and will likely kill them. A lower temperature basking spot can be Development and Innovation, provided with incandescent lamps Queensland. 16pp. positioned at one end of the enclosure. The temperature on the branching Daniels, R, R, J. 2005. Amphibians below the basking spot should not on peninsular India. Universities o exceed 28 C. Small tanks will rapidly press, Hyderabad. 288pp overheat so ensure that there is still a cool area in the enclosure. Metamorphs should be fed daily until about six weeks Fan, X., Lin, Z. & Ji, X. (2013) Male of age with pin head crickets and size does not correlate with (occasionally) Drosophila. All food fertilization success in two bufonid items should be dusted with appropriate toads that show size assertive dietary supplements. After 8 weeks the mating. Current Zoology 59 (6): feeding interval can be gradually 740-746. increased to once every 3 days. IUCN, Conservation International, Health and NatureServe. 2006. Global A m p h i b i a n A s s e s s m e n t . Problems Encountered in Captivity . Toads are often sit and wait predators, Accessed on 09 October 2007. their calorific intake often exceed their need and obesity is common in captive Mathew, R. & Sen, N. (2010) toads. Pictorial guide to amphibians of North East India. Zoological Survey Record keeping of India, Kolkata. 144pp.

Things to include. Individual identification, weight, feeding records, temperature records, UV records, water quality test results, health records, behavioural or reproductive observations.

10 Ex situ Management of Amphibians

3. Ex-Situ Management of Microhylid frogs Kaloula, Microhyla, Ramanella & Uperodon species Distribution R a m a n e l l a v a r i e g a t a , K a l o u l a taprobanica, Uperodon ayatoma, Microhyla ornata and Microhyla rubraare distributed over much of peninsular India. Kaloula pulchra occurs in , Southern

1 China and North East India (IUCN et al., 2006).

Conservation Status and Threats All of the above species are listed as Least Concern (IUCN et al., 2006).

Habitat and Ecology

2 Ramanella variegata (Fig 1). can be found in various types of forest, grassland and pastures as well as urbanised areas. It is found in dry soil areas with ground cover. They are often found in association with mounds in India (IUCN et al., 2006). They are explosive breeder in temporary 3 pools and breeding starts with the onset of the monsoon rains (Daniels, 2005).

Kaloula pulchra are commensal with humans and can rapidly colonise urban habitats. It is adaptable and was presumably originally a wetland 4 /riverbank/forest edge. It can still be Figure 1. Top Ramanella variegata (Sandeep Varma). found in dry and evergreen forests. It is a Figure 2. Middle top Kaloula taprobanica (Sandeep Varma) nocturnal and fossorial species and Figure 3. Middle bottom (Sandeep Varma) breeds in small pools (IUCN et al., Figure 4. Bottom Microhyla rubra (Sandeep Varma) 2006). Kaloula are able to climb well when looking for shelter (Daniels, Introduction 2005).

Description Kaloula taprobanica (Fig.2). is a fossorial, nocturnal species which can be Indian Microhylid frogs (Fig.1) are found in dry forests, coconut and rubber characterised by the small to medium plantations, wetlands and areas close to size, small heads, invisible ear human habitations. It is most often (Typmanum) ad squat body (Daniels, found under leaf-litter, in loose soil, and 2005). 11 Ex situ Management of Amphibians under logs and other ground cover. lay egg masses Breeding and larval development take which float on the water surface (IUCN place in pools (IUCN et al., 2006).In et al., 2006). Whilst mating the pair glue Chennai this species breeds from themselves together with a sticky October-November (Daniels, 2005) secretion, eggs measure 1-1.5mm in diameter when they are laid (Daniels, Uperodon (Fig. 3). are completely 2005). fossorial that burrow in loose, moist soil up to a depth of 2 meters (Daniels, Uperodon systoma lay egg masses 2005). They have been observed in dry which float on the water surface at the forest areas, plains, home gardens and onset of the monsoon rains (IUCN et al., agricultural areas. Adults surface during 2006). the summer monsoons. Breeding occurs during the monsoon rains. Males call W h i l s t m a t i n g M . o r n a t a g l u e from the banks of torrents or paddy themselves together with a sticky fields (IUCN et al., 2006). In Pakistan, secretion, up to 1,300 eggs are laid by termites are reportedly the main food of each female (Fig. 8), the eggs are 0.4mm this species. when laid and take 48 hours to hatch. The tadpoles are plankton feeders and Microhyla ornata and M. rubra (Fig. 4). metamorphose in 2-3 weeks (Fig. 7). inhabit dry forest, shrub land, grassland, agricultural land and urbanised areas. They can found in loose soil, amongst leaf-litter, and under logs and other ground cover. Breeding takes place in still waters and paddy fields. (IUCN et al., 2006). They are nocturnal but can also be active during the day in the breeding season. Microhyla breed during the onset of the monsoon rains (Daniels, 2005) 5

Sexing Individuals Microhylid frogs can be difficult to sex, females tend to be larger than the males.

Amplexus, Egg Laying and Larval development Ramanella variegata is an explosive 6 breeder in temporary pools. The tadpoles are bottom dwellers and metamorphosis is thought to occur in less than a month (Daniels, 2005).

7 Kaloula pulchra breed in pools (IUCN et Figure 5. Top Spawning K. pulchra (Benjamin Tapley) al., 2006). Larval development take Figure 6. Middle. K. pulchra tadpole (Benjamin Tapley) place in still pools (Fig 5 & 6). Figure 7. Microhyla tadpole (Sandeep Varma) In India the reproductive biology of Kaloula taprobanica breed in pools Microhyla rubra has not been studied in ( I U C N e t a l . , 2 0 0 6 ) . T a d p o l e s great detail but in Sri Lanka egg clutches metamorphose 30-45 days after the eggs consisted of 400 eggs and tadpoles took are laid (Daniels, 2005). 77 days to metamorphose at 22-24oC (Bowatte & Meegaskumbura, 2011). 12 Ex situ Management of Amphibians

Longevity and Age at Sexual should be deep enough for the frogs to Maturity immerse their drink patches. Lifespan in the wild is unknown. Cover objects are essential when Captive Management keeping terrestrial frogs. 150 mm lengths of PVC pipe or hollowed bamboo make appropriate cover objects, these Introduction need to be able to accommodate the Kaloula pulchra are established in frogs so the size of the tube will depend captivity but are infrequently bred. This on the size of the frog. Live plants can be species was bred at Durrell but many of added to the enclosure as they create the eggs were infertile and the 12 humid pockets which the animals may individuals that metamorphosed died use, the plants also provide visual less than two weeks after hatching barriers for the frogs. despite feeding and growing. Daniels (2005) reports that an attempt to Lids should be constructed of mesh as maintain U. systoma in captivity failed. glass will block UVB.

Substrate must be provided for this species as they all dig. The provision of substrate will prevent the enclosure from drying out. Be sure to incorporate a drain in the tank construction. A layer of gravel covered with a permeable matting should be laid underneath a substrate of soil mixed with sand (ratio 2:1). The depth of substrate will depend on enclosure size but must be at least 150 mm deep for Kaloula, Ramanella and Microhyla and at least 300 mm for Uperodon. The larger the enclosure, the Figure 8. Developing Microhylid eggs (Benjamin Tapley) deeper the substrate should be as Identifying Individuals substrate will dry out quickly in large Adult specimens of all species have enclosures. Substrate does not need to distinctive markings on the dorsal be compacted for this species. Ensure surface, photographic identification that substrates are not contaminated would be appropriate for adults of this with fertilisers or other substances that species. may be toxic to amphibians.

Housing adults Ensure the substrate has a humidity gradient by pouting some water (or A floor space of 600 x 600 mm would be spraying with water) more at the end of the minimum enclosure size for 5 the enclosure with no basking light. individuals of Kaloula and Uperodon These are litter frogs and are well species. 8 Microhyla or Ramanella camouflaged, a layer of leaf litter should could be maintained in this size be provided for these frogs as they may enclosure. Height is not important as experience some level of stress without these frogs rarely climb. The height of it. the enclosure should allow them to hop freely without coming into contact with the lid, an enclosure height of 700 mm Temperature, Humidity and would be sufficient. A wire mesh lid Lighting should be provided. A 200 x 150 x 100 Husbandry should be informed by data mm water dish should be provided and collected from the field. Basking spot 13 Ex situ Management of Amphibians temperatures, UV exposure and and refilled daily as amphibians often gradients, temperature and humidity excrete waste products in water. Water r e g i m e s s h o u l d m i m i c t h e quality cannot be judged by eye. Faeces environmental conditions at the and uneaten food must be removed collection site. Note that environmental daily. The enclosure and all furniture data should be collected at different must be thoroughly scrubbed (with a times throughout the day and year. brush and hot water, no chemical Suitable ambient room temperatures cleaners or disinfectants) twice a week. should vary between 23 and 27˚C Temperatures of the enclosure should be (night/day summer) and 20 and 25˚C checked at different points of the (night/day winter). enclosure regularly to ensure that the correct thermoregulatory gradients are A UV gradient should be provided in maintained. UV readings must be taken conjunction with a basking spot. Arcadia every three months to check that there is D3+ reptile lamp T5 12% UVB are ideal a sufficient UV gradient and that the UV and a maximum UVI of 3 should be emitting lights are still emitting UV. provided for the frogs at their basking spots. Remember that mesh will filter The substrate must be changed as soon out some of the UVB so UV readings as it looks dirty or if it becomes saturated must be taken from below the mesh with or unable to retain humidity. a solarmeter 6.5. Diet in captivity Basking spots can be provided with Uperodon are reportedly termite regular incandescent lamps. The specialists (Daniels, 2005; IUCN et al., basking lamp should be positioned at 2006). Kaloula will readily accept small one end of the enclosure and the live insects in captivity. A study on the temperature on the substrate below the diet of wild Microhyla heymonsi in basking spot should range between 30- Indonesia showed that more than 50% 32oC and not exceed 32oC. Ensure that all of the diet is made up of ants but they lighting is inaccessible to the frogs as consume a wide variety of small they will be burnt easily if they come into i n v e r t e b r a t e s ( E r f t e m e i j e r & contact with lamps. Boeadie,1991). Great care should be taken when feeding captive amphibians Check the source of your water by testing ants as ants can be aggressive and may it regularly for ammonia, nitrite, nitrate, bite and sting the toads, this is pH and phosphates. Tap water should be problematic because the toads cannot left to stand for 24 hours before it is used escape the ants if they choose to. The in order for the chlorine to dissipate. feeding habits of Ramanella are Ensure the temperature of the water is at unknown, like other similar sized frogs equilibrium with the room temperature. they probably feed on invertebrates. A plastic water dish should be provided Food should be offered after it is dusted and should measure 200 x 150 mm. with an appropriate dietary supplement, Water depth should be 80 mm, and frogs well fed and an appropriate size. should be able to immerse their drink Microhylids have small mouths so the patch. An emergent rock should be food should be no greater in size than the placed in the water to allow the frogs distance between the yes. of the frogs easy access out of the water. The head. Microhylid frogs are usually enclosure should be lightly misted with nocturnal and should be fed in the aged tap water daily. evening.

Routine Husbandry Adults should be fed every three to six All animals should be visually inspected days (depending on season and daily. Water dishes must be cleaned out condition), juveniles up to six weeks of

14 Ex situ Management of Amphibians age should be fed daily. could be used (Appendix 1). Water depth is extremely important when breeding Reproduction in Captivity microhylids. A gradient should always be provided. During breeding Kaloula Breeding Enclosure and Uperodon will duck under the Increasing the humidity may trigger surface during egg laying and the water breeding in this species. These frogs depth needs to be deep enough to allow should be cycled in a rain chamber with a this to happen. The water depth should sprinkler system for breeding (Fig. 9). be 150 – 200 mm at one end of the rain chamber. The, lighting and temperature A rain chamber with a floor space of 600 parameters should be maintained as x 1000 mm would be suitable as a above. Visual barriers such as palm minimum size for 2 males and 3 females fronds or plastic plants could be used to (Kaloula and Uperodon) or 4 males and provide cover for the frogs in their 4 females (Microhyla and Ramanella). It breeding enclosure if they fail to spawn is important that the sex ratio in the after a couple of weeks. breeding enclosure is not biased towards males this may be stressful for the The frogs should be fed at least every females). No substrate is required in the other day whilst they are in the rain rain chamber at first, but a 1cm deep chamber but ensure that any dead insects are removed daily as they will pollute the water.

Never try and separate Microhylids once they begin mating as many species glue themselves together during the process, pulling them part could damage their skin.

In closed systems regular water tests should be carried out (Temperature, pH, ammonia, nitrite and nitrate), the results will help you decide the frequency of water changes. pH should not fluctuate, ammonia should measure less than 1 part per million (ppm), nitrite less than 0.1ppm and nitrate less than 10ppm. No more than 20% of the water should be changed as rapid shifts in Figure 9. A rain chamber suitable for breeding C. curtipes water quality will be problematic for the (Benjamin Tapley) biological filtration process. lining of nontoxic clay could be trialled if the frogs have not spawned after one Breeding Seasonality month. Based on their morphology most Breeding aggregations of these species microhylids are poor swimmers so it is can be found at the start of the monsoon. vital that there are some rocks or logs in the water that the frogs can easily leave Provision of Breeding Sites the water should they choose to do so. A A depth gradient should be provided for pump powered spray system or fountain the frogs so that they can choose their would be ideal for breeding this species egg laying site, this can be provided by but a mature biological filter should be angling slates or paving slabs into the incorporated into the rain chamber to water maintain water quality. Ideally a sump 15 Ex situ Management of Amphibians

Egg Care, Tadpole Husbandry When the eggs first hatch the tadpoles & Development will not move and it is perfectly normal When eggs are laid it is best to leave for them to sometimes rest on their side. them in the rain chamber and remove If you look on the underside of tadpoles the frogs, many microhylid eggs float on when they first hatch you should be able the surface of the water and moving to see the yolk sac. The tadpole will not them may make them sink, it is start swimming or feeding until this ac unknown what impact tis would have on has been absorbed. Once the newly egg development. You may not wish to hatched tadpoles have dropped off the keep all the eggs if the clutch size is large egg mass it is important to remove the as rearing large numbers of tadpoles and remaining egg jelly as this will decay and providing food for all the metamoprhs pollute the water. Substrate is not will be a huge task. necessary but tadpoles like to hide. Dried Indian almond leaves (Terminalia The size of the tank is not important, the catappa) can be added, they also soften stocking density is. Tanks should be the water which would be beneficial in stocked with no more than 5 tadpoles hard water areas. Partial (10-20 %) per litre.. Filtration is vital (see appendix water changes should be carried out one to inform your choice of filter). Aged regularly, the frequency of water tap water can be used to rear tadpoles. changes must be governed by frequent Check the source of your water by testing water testing. it regularly for ammonia, nitrite, nitrate, hardness, pH and phosphates. Water Tadpoles should be fed daily, once they parameters should be informed by field start free swimming. They should be fed data but remember that not all breeding on a variety of food. At Durrell Kaloula sites may have optimal water quality. tadpoles were reared to metamorphoses Tap water should be left to stand for 24 o n a p o w d e r e d t a d p o l e f o o d hours before it is used in order for the (components : ground tropical fish flake, chlorine to dissipate. Ensure the grass pellet, trout pellets, tubifex, river t e m p e r a t u r e o f t h e w a t e r i s a t shrimp, spirulina algae and cuttlefish equilibrium with the room temperature. bone). Food should be available to the Check the water in closed systems and tadpoles at all times but it is important c a r r y o u t r e g u l a r w a t e r t e s t s that large amounts of uneaten food do (temperature, pH, ammonia, nitrite and not pollute the aquarium. Tadpole nitrate), the results will help you decide feeding regimes must be modified, as the the frequency of water changes. pH tadpoles grow they will eat more. should not fluctuate, ammonia should measure less than 1 part per million Once the front limbs appear and the tail (ppm), nitrite less than 0.1ppm and starts to reduce it is important that the nitrate less than 10ppm. No more than metamorphosing froglets are given the 20% of the water should be changed as opportunity to emerge from the water as rapid shifts in water quality will be they can easily drown. Emerging rocks problematic for the biological filtration or aquatic plants are suitable. Once they and tadpoles. Ideally a sump should be emerge they are prone to drying out and used to increase the volume of water of easily die if the humidity is not high. your system and to house all the life support systems (filters etc). If using an Rearing Metamorphs internal canister filter ensure that it is not too strong as the tadpoles could be sucked into the inflow of the filter and may have difficulty feeding. Direct the outflow to the surface of the water for oxygenation. Figure 10. Enclosure for rearing juvenile Micohylids (Benjamin Tapley). 16 Ex situ Management of Amphibians

Metamorphs can be housed in small rapidly and will likely kill them. A lower plastic containers. A 350 x 200 x 300 temperature basking spot can be mm enclosure is suitable for up to 12 provided with incandescent lamps individuals (Fig. 10), these groups positioned at one end of the enclosure. should be divided amongst larger The temperature on the branching enclosures as individuals grow. It is below the basking spot should not important that rearing tanks are exceed 28oC. Small tanks will rapidly relatively small so that high ambient overheat so ensure that there is still a humidity can me maintained and that cool area in the enclosure. Metamorphs the juvenile frogs can find food easily. should be fed daily until about six weeks Covering the lid with a fine breathable of age with pin head crickets and cloth or fleece will prevent the food (occasionally) Drosophila. All food escaping and ensure that humidity items should be dusted with appropriate remains high. The cloth will block dome dietary supplements. After 8 weeks the of the UV emitted from the lights so you feeding interval can be gradually must experiment and ensure that the UV increased to once every 3 days. light and temperature gradients are still suitable. Compacted damp soil mixed Health with sand would make a good substrate, and this must be replaced as necessary. Problems Encountered in It is important that the substrate is Captivity Obesity. compacted when housing metamorphs as the soil particles easily stick to the Record keeping skin of the froglets. A shallow water dish must be provided (water depth 10 mm). Things to include. It is essential that the water dish is easy for the metamorphs to climb out of (a Individual identification, weight, rock or small branch in the water dish feeding records, temperature records, facilitates access). When transferring UV records, water quality test results, froglets from the tadpole tank make sure h e a l t h r e c o r d s , b e h a v i o u r a l o r that they are placed in the water dish and reproductive observations. not on the substrate as they may not be quite ready to leave the water, they will Bibliography choose when they are ready. Cover Bowatte, G.. & Meegaskumbura, M. objects are extremely important 2011. Morphology and ecology of microhabitats for small toads. Bark Microhyla rubra (Anura: Microhyidae) hides and leaf litter should be added to tadpoles from Sri Lanka. Amphibian and the enclosure. The furnishings in the reptile conservation 5 (2): 22-32. enclosure should not be too complex or it will be difficult for the young frogs to Daniels, R, R, J. 2005. Amphibians on find food. The metamorphs can be raised peninsular India. Universities press, at the same room temperature as the Hyderabad. 288pp adults (23 - 27˚C), and must be misted with room temperature tap water twice Erftemeijer, P. & Boeadie (1991) The diet daily. It is vital that metamorphs and o f M i c r o h y l a h e y m o n s i , V o g t juveniles are provided with UV at this () and chalconota, critical growing stage (Arcadia D3+ Schlegl (Ranidae) in a pond on West reptile lamp T5 12% UVB), this must be Java. Raffles Bulletin of Zoology 39 (2): given in a gradient with a maximum UVI 279-282. of 2 (Fig. 9). IUCN, Conservation International, and Do not provide metamorphs with a 32oC Nature Serve. 2006. Global Amphibian t basking spot as this will dry them out Assessment.. Accessed on 09 October 2007. 17 Ex situ Management of Amphibians

4. Ex-situ Management of Leaping frogs Indirana species

conservation status of Indirana is summarised in table 1. Species IUCN status Distribution range (km2) Indirana beddomii Least concern 1250 Indirana brachytarsus Endangered 550 Indirana semipalmata Least concern 700 Indirana leithii Vulnerable 500 Indirana leptodactyla Endangered 600 Indirana diplosticta Endangered 350 Indirana gundia* Critically 50 endangered Indirana phrynoderma Critically 100 endangered Inditana longicrus* Data deficient 50 Indirana tenuilingua* Data deficient 50

*Known from only one locality. Table 1. Conservation status of Indirana (Nair et al., 2012)

Habitat and Ecology Indirana frogs are ground dwelling and live on the forest floor of evergreen and semi deciduous forests, usually in association with streams with wet rock faces (IUCN et al., 2006; Tapley et al., Figure 1. Indirana leptodactyla (Benjamin Tapley). 2011; Nair et al., 2012). Indirana breed during the monsoon rains (Daniels, Introduction 2005). Vocalisations consist of short calls, males of some species call Description simultaneously (Kadadevaru et al, Frogs of the genus Indirana (Family 20o0; Kuramoto & Dubois, 2009) ) can be distinguished from suggesting males may form loose leks most other sympatric frogs by the (Nair et al., 2012) following characteristics small in size (less than 80 mm snout vent length Sexing Individuals (SVL), brown in colouration, prominent Females are usually larger than the tympanum (ear), fingers and toes males. Relative tympanum size is terminating in discs, round pupil greater in male frogs (Nair et al., 2012). (Daniels, 2005). Male Indirana in breeding condition have nuptial pads on the first finger Distribution (Daniels, 2005; Gopalan, et al., 2012; Indirana are endemic to the Western Nair et al., 2012). Ghats Mountains of south-western India (Daniels, 2005) and the genus has an altitudinal range from 0 - 2,000 masl (Nair et al., 2012).

Conservation Status and Threats Several species re known only from their type localities (IUCN et al., 2006). The Figure 2. Eggs of I. semipalmata (Benjamin Tapley) 18 Ex situ Management of Amphibians

Egg Laying and Larval Captive Management development Indirana lay their eggs in humid areas in Introduction clear poos at the base of rocks or in rock Indirana is not currently kept in crevices (Daniels, 2005). Indirana captivity. bedomii is reported to lay eggs in shallow pools (20 mm in depth) away Identifying Individuals from streams (Veeranagoudar et al., Not all Indirana possess patterns 2009). Indirana leptodactyla is reported suitable for photographic identification. to lay up to 30 eggs per clutch in the Adults of the larger species could be grassy margins of ponds (Rao, 1920). marked Adults could be marked with Tapley, et al., reported that I. passive integrated transponders (PIT semipalmata lay clumps of up to 343 tags). Pit tags should be inserted eggs on sheltered wet rock faces and subcutaneously on the dorsum. even on tree bark (Fig 2.). Juveniles could be marked with different coloured visible implant Indirana tadpoles (Fig. 3). are elastomer (VIE) but this may not be extremely specialised, they have long necessary for rearing purposes. tails with reduced fins which suit their specialised locomotion on usually Housing adults vertical wet rock surfaces and even on Indirana require specialist housing due wet tree bark away from any permanent to their unique microhabitats. An streams (Veeranagoudar et al., 2009; enclosure measuring 600 x 6000 x 600 Tapley et al, 2011). Tadpoles are exposed mm would be the minimum enclosure to visually orientated predators in such size for 8 individuals (Fig.4). A wire environments and rely on camouflage, mesh lid should be provided. Water early hind limb development and should be provided in two forms, a pool muscular tails which allow them to make and a wet rock surface. quick jumps when disturbed (Annandle, 1918, Kuramoto & Joshy, 2002; Substrate must be provided for this Veeranagoudar et al., 2009; Tapley et al, species as it is unknown whether or not 2011 ). The incredible jumping ability of they like to dig. The provision of Indirana tadpoles allow them to escape substrate will prevent the enclosure microhabitats which may dry out from drying out. Be sure to incorporate a rapidly. drain in the tank construction. A deep layer of gravel (200 mm) covered with a permeable matting should be laid underneath a substrate of soil mixed with sand (ratio 2:1). The depth of the sands / soil substrate should be at approximately 50 mm deep. Ensure that substrates are not contaminated with fertilisers or other substances that may be toxic to amphibians.

Ensure the substrate has a humidity gradient by pouting some water (or spraying with water) more at the end of Figure 3. Typical Indirana tadpole (Benjamin Tapley) the enclosure with no basking light. Longevity and Age at Sexual These frogs and are well camouflaged, a Maturity layer of leaf litter should be provided for Lifespan in the wild is unknown. these frogs as they may experience some level of stress without it.

19 Ex situ Management of Amphibians

Cover objects are essential when from feeding and may wash them off the keeping terrestrial frogs. Plants can be rock surface. added to the enclosure as they create humid pockets which the animals may Temperature, Humidity and use, the plants also provide visual Lighting barriers for the frogs. Small lengths of Husbandry should be informed by data PVC pipe or hollow bamboo may also be collected from the field. Basking spot beneficial. Lids should be constructed of temperatures, UV exposure and mesh as glass will block UVB. gradients, temperature and humidity r e g i m e s s h o u l d m i m i c t h e environmental conditions at the collection site. Note that environmental data should be collected at different times throughout the day and year. Suitable ambient room temperatures should vary between 23 and 27˚C (night/day summer) and 20 and 25˚C (night/day winter).

A UV gradient should be provided in conjunction with a basking spot. Arcadia Figure 4. Set up suitable for housing and breeding Indirana D3+ reptile lamp T5 12% UVB are ideal (Brian's Tropicals) and a maximum UVI of 3 should be The water pool van be created by by provided for the frogs at their basking providing a large indentation in the spots. Remember that mesh will filter gravel (Fig. 4). The sides of the pool can out some of the UVB so UV readings be maintained by growing live plants on must be taken from below the mesh with the substrate, or using pebbles or bricks a solarmeter 6.5. to create the edge of the pool. Filtration must be incorporated into this design to Basking spots can be provided with maintain water quality. In closed regular incandescent lamps. The systems regular water tests should be basking lamp should be positioned at c a r r i e d o u t ( T e m p e r a t u r e , p H , one end of the enclosure and the ammonia, nitrite and nitrate), the temperature on the substrate below the basking spot should range between 30- results will help you decide the o o frequency of water changes. pH should 32 C and not exceed 32 C. Ensure that all not fluctuate, ammonia should measure lighting is inaccessible to the frogs as less than 1 part per million (ppm), nitrite they will be burnt easily if they come into less than 0.1ppm and nitrate less than contact with lamps. These frogs can 10ppm. No more than 20% of the water often be found on dappled forest floor should be changed as rapid shifts in during the day (Pers. obs) and so the water quality will be problematic for the provision of UV lighting is essential. biological filtration process. A wet rock face can be created by securely angling a Lighting is also important for the growth slate or other flat rock surface against of alga on the flat rock surface one side of the enclosure with the (Important tadpole food). bottom of the slate being firmly positioned in the pool of water (Fig. 4). A Check the source of your water by submersible pump with adjustable flow testing it regularly for ammonia, nitrite, can be buried in the gravel. The so that nitrate, pH and phosphates. Tap water water trickles down the rock face. It is should be left to stand for 24 hours important flow is adjustable, a powerful before it is used in order for the chlorine stream of water will prevent tadpoles to dissipate. Ensure the temperature of 20 Ex situ Management of Amphibians the water is at equilibrium with the room Reproduction in Captivity temperature. Breeding Enclosure Increasing the humidity may trigger The enclosure should be lightly misted breeding in this species. These frogs with aged tap water daily. should be cycled in their regular enclosure for breeding (Fig. 4). Routine Husbandry All animals should be visually inspected Breeding Seasonality daily. Dead food items and faecel There is no published report on the material must be removed from the captive breeding of Indirana. In the water daily as amphibians often excrete wild, breeding aggregations of this waste products in water. Water quality species can be found at the start of the cannot be judged by eye. Faeces and monsoon. uneaten food must be removed from the enclosure daily. The enclosure and all Provision of Breeding Sites furniture must be thoroughly scrubbed A depth gradient should be provided for (with a brush and hot water, no chemical the frogs so that they can choose their cleaners or disinfectants) twice a week egg laying site, this can be provided by but do not scrub the wet rock surface as angling slates or paving slabs into the algae will grow on it and this is the most water (Fig.4). important food for the tadpoles. Temperatures of the enclosure should be Egg Care, Tadpole Husbandry & checked at different points of the Development enclosure regularly to ensure that the Leave the eggs in the same enclosure as correct thermoregulatory gradients are the parents, I. semiplamata often sit maintained. UV readings must be taken beside their egg clutches and the skin of every three months to check that there is the adult frogs could have anti fungal a sufficient UV gradient and that the UV properties which prevent the eggs emitting lights are still emitting UV. getting mouldy. Some frogs are also known to eat frog eggs, egg guarding by The substrate must be changed as soon adult individuals could prevent this as it looks dirty or if it becomes saturated from occurring. Eggs are sometimes laid or unable to retain humidity. in quite exposed places and they could easily dry out. A fogger should be placed Diet in captivity in aquarium to maintain high ambient The feeding habits of these frogs are humidity. The fogger can be connected largely unknown, I. semipalmapata to a timer, and turned on for two minutes have been observed feeding on termites every hour. You may need to adjust this (pers obs). Like other similar sized frogs if the enclosure becomes too wet or eggs they probably feed on invertebrates. still appear to dry out. Food should be offered after it is dusted with an appropriate dietary supplement, When the eggs first hatch the tadpoles well fed and an appropriate size, no will not move and it is perfectly normal greater in size than the width of the frogs for them to sometimes rest on their side. head. Indirana are nocturnal and If you look on the underside of tadpoles should be fed in the evening. when they first hatch you should be able to see the yolk sac. The tadpole will not Adults should be fed every three to six start feeding until this ac has been days (depending on season and absorbed. condition), juveniles up to six weeks of age should be fed daily. Tadpoles feed on wet rock surfaces and maintain the tadpoles is probably the most challenging aspect of Indirana

21 Ex situ Management of Amphibians husbandry. The rock surface may rock or small branch in the water dish become depleted of alga very quickly. It facilitates access). When transferring is therefore important that several spare froglets from the tadpole tank make sure rock surfaces are prepared ahead of that they are placed in the water dish and planned breeding trials. These can be set not on the substrate as they may not be up in an adjoin well lit aquarium with a quite ready to leave the water, they will similar water fall like system. As a large choose when they are ready. Cover resort suitable rocks could be removed objects are extremely important from the field. Great care should be microhabitats for small frogs. Bark hides taken when working with enclosures and leaf litter should be added to the with tadpoles as you don't want the enclosure. The furnishings in the tadpoles to jump off the rock as they may enclosure should not be too complex or not be able to find the rock and food it will be difficult for the young frogs to source easily once they leave it. Any find food. The metamorphs can be raised movement incursions into the tank at the same room temperature as the should involve slow movements and the adults (23 - 27˚C), and must be misted reaction of the tadpoles to the keeper with room temperature tap water twice closely observed. Do not remove the daily. It is vital that metamorphs and existing rock if more food needs to be juveniles are provided with UV at this added, simply place the new rocks on or critical growing stage (Arcadia D3+ beside the existing rock surface, making reptile lamp T5 12% UVB), this must be sure that they too are covered by a trickle given in a gradient with a maximum UVI of water. of 2 (Fig. 9).

Rearing Metamorphs Do not provide metamorphs with a 35oC Once the tadpoles metamorphose (tail basking spot as this will dry them out totally absorbed) they can be housed in rapidly and will likely kill them. A lower small plastic containers. A 350 x 200 x temperature basking spot can be 300 mm enclosure is suitable for up to 12 provided with incandescent lamps individuals (Fig. 9), these groups should positioned at one end of the enclosure. be divided amongst larger enclosures as The temperature on the branching individuals grow. It is important that below the basking spot should not rearing tanks are relatively small so that exceed 28oC. Small tanks will rapidly high ambient humidity can me overheat so ensure that there is still a maintained and that the juvenile frogs cool area in the enclosure Metamorphs can find food easily. Covering the lid should be fed daily until about six weeks with a fine breathable cloth or fleece will of age with springtails, pin head crickets prevent the food escaping and ensure and (occasionally) Drosophila. All food that humidity remains high. The cloth items should be dusted with appropriate will block dome of the UV emitted from dietary supplements. After 8 weeks the the lights so you must experiment and feeding interval can be gradually e n s u r e t h a t t h e U V l i g h t a n d increased to once every 3 days. temperature gradients are still suitable. Providing small enough prey items for Compacted damp soil mixed with sand Indirana could be very difficult as the would make a good substrate, and this f r o g l e t s a r e t i n y w h e n t h e y must be replaced as necessary. It is metamorphose. It is important that important that the substrate is adequate stocks of live food are available compacted when housing metamorphs well in advance of breeding. as the soil particles easily stick to the skin of the froglets. A shallow water dish Health must be provided (water depth 10 mm). It is essential that the water dish is easy Indirana leithi in Maharashtra have for the metamorphs to climb out of (a tested positive for Batrachochytrium

22 Ex situ Management of Amphibians dendrobaitidis, the fungus which causes Ghats, India. Herpetology Notes. 5: 263- t h e d i s e a s e c h y t r i d i o m y c o s i s 273. (Dahanuker et al., 2013) IUCN, Conservation International, and NatureServe. 2006. Global Amphibian Problems Encountered in Assessment.. Accessed on 09 October 2007. Non applicable. Kadadevaru, G.G., Kanamadi, R.D. & Record keeping Schneider, H. 2000. Advertisement calls in two Indian ranids, Indirana bedomii Things to include. and Tomopterna rufescens. Amphibia- Individual identification, weight, Reptilia 21: 242-246. feeding records, temperature records, UV records, water quality test results, Kuramoto, M. & Dubois, A. 2009. h e a l t h r e c o r d s , b e h a v i o u r a l o r Biacoustic studies on three frog species reproductive observations. from the Western Ghats, South India. Current Hereptology 28: 65-70. Bibliography Kuramoto, M., Joshy, S.H. 2002. Annandale, N. 1918. Some undescribed Tadpoles of Indirana beddomii (Anura: tadpoles from the hills of southern India. Ranidae). Hamadryad 27: 71-77. Recordings of the Indian Museum, Calcutta15: 17-24. Nair, A., Gopalan, S.V., George, S., Kumar, K.S., Techer, A.G. & Merilä, J. Dahanuker, N., Krutha, k., Paingakar, 2012. Endemic Indirana frogs of the M.S., Padhye, A.D., Modak, N. & Molur, Western Ghats biodiversity hotspot S. 2013. Endemic Asian chytrid strain Ann. Zool. Fennici. 49: 257-286. infection in threatened and endemic anurans in the Northern Western Ghats, Tapley, B., Purushotham, C.B., Girgin, S. India. Plos one 8(10): e77528. 2011. Indirana semipalmata (Brown l e a p i n g f r o g ) R e p r o d u c t i o n . Daniels, R, R, J. 2005. Amphibians on Herpetological Review. 42: 87-88. peninsular India. Universities press, Hyderabad. 288pp Veeranagoudar, D.K., Radder, R.S., Shanbhag, B.A. & Saidapur, S.K. 2009. Gopalan, S.V., Nair, A., Santosh Kumar, Jumping behaviour of semi terrestrial K., Merilä, J. & George, S. 2012. tadpoles of Indirana beddomii Morphology of Indirana semipalmata (Günther): relative importance of tail (Boulenger, 1882) (Amphibia; Anura) and body size. adults and tadpoles from the Western

23 Ex situ Management of Amphibians

5. Ex-situ Management of Bicoloured frog Clinotarsus Curtipes

deciduous forest. It is also found in lightly degraded areas (IUCN et al., 2006). The abundance of C. curtipes is related to large water bodies and canopy-covered habitats (Krishna & Krishna, 2005).

Sexing Individuals Clinotarsus curtipes are sexually dimorphic with females much larger Figure 1. Clinotarsus curtipes (Benjamin Tapley). than males (Fig. 2).

Introduction

Description Bicoloured frogs are stocky bodied and thin limbed ranid frogs (Fig.1) C. curtipes is polymorphic, colouration is highly variable, even amongst wild specimens collected in the same areas. Female The tips of the fingers and toes terminate in small discs, the toeas are fully webbed (Daniels, 205). Clinotarsus Figure 2. Amplectant C. curtipes, note the small size of the curtipes may grow to 75 mm snout vent male (Benjamin Tapley) length (SVL). Reproduction and Larval Development Distribution These frogs breed in deep, perennial Clinotarsus curtipes are endemic to the water bodies such as along slow-flowing Western Ghats Mountains of south- streams and in water tanks with canopy western India (Daniels, 2005) from 500 cover and dams (Hiragond et al., 2001, - 2,000 mask (IUCN et al., 2006). Krishna and Krishna, 20051,2) Males congregate around slow moving or still Conservation Status and Threats water bodies where they vocalise Clinotarsus curtipes is listed as Near (Daniels, 2005). Large numbers of frogs Threatened by the IUCN Red List as its congregate around breeding sites after extent of occurrence is approximately dusk and emit their advertisement calls 20,000 km2, and the extent and quality from 1820hours until 0400hrs. Female of its habitat are probably declining, C.curtipes have been documented thus making the species close to vocalising from the same location at a qualifying for Vulnerable. (IUCN et al., breeding site over multiple nights 2006). Tadpoles of this species are (Krishna and Krishna, 20051). Females collected for human consumption who were still vocalising late in the (Stuart et al., 2008) breeding season and were holding territories were observed fighting and Habitat and Ecology chasing off intruding females who Clinotarsus curtipes is a terrestrial leaf- arrived late at the breeding site and such litter frog, found in evergreen to semi- fights were not recorded among females evergreen moist forest, and into dry who arrived early in the breeding season

24 Ex situ Management of Amphibians in the presence of many males (Krishna and Krishna, 2005 1 ). Males are territorial and have been fighting during t h e b r e e d i n g s e a s o n ( T a p l e y & Purushotham, 2011). Both males and females are known to vocalise at the breeding sites (Krishna & Krishna, 2005 1).

Amplexus, Egg Laying and Larval development Amplexus (mating position) is axillary in this species. (Daniels, 2005).It is t h o u g h t t o b e e g g l a y i n g , t h e reproductive biology of this species is not well studied. The tadpoles of Figure 5. The mottled underside of C. curtipes (Benjamin Tapley). C.curtipes are thought to feed on alga and are amongst the largest tadpoles in Captive Management the Western Ghats and can attain a total length of 9 cm (Fig 3.). Tadpoles have Introduction three glands on the dorsal surface which Clinotarsus curtipes is not currently excrete a white fluid when handled, it is kept in captivity. not known whether or not this fluid is toxic (Fig 4.). These tadpoles form large shoals and kin recognition is known in Identifying Individuals this species, each shoal being composed Adult specimens of both species have of siblings (Gururaja KV pers.com). distinctive markings on the ventral s u r f a c e ( F i g . 5 ) , p h o t o g r a p h i c identification would be appropriate for adults of this species.

Housing adults Clinotarsus curtipes require relatively large enclosures, a floor space of 600 x 1000 mm would be the minimum enclosure size for 5 individuals (Fig.6). Height is not important as these frogs 3 rarely climb. The height of the enclosure should allow them to hop freely without coming into contact with the lid, an enclosure height of 500 mm would be sufficient. A wire mesh lid should be provided. A 250 x 150 x 100 mm water dish should be provided and should be 4 deep enough for the frogs to immerse Figure 3. Top. A shoal of C. curtipes larva, note the orange their drink patches. glands (Chetana Purushotham) Figure 4. Maximum tadpole size of C. curtipes (Benjamin Tapley) Cover objects are essential when keeping terrestrial frogs. 150 mm Longevity and Age at Sexual lengths of PVC pipe or hollowed bamboo Maturity make appropriate cover objects, these Lifespan in the wild is unknown. need to be able to accommodate the frogs so the size of the tube will depend 25 Ex situ Management of Amphibians on the size of the frog. Live plants can be Temperature, Humidity and added to the enclosure as they create Lighting humid pockets which the animals may Husbandry should be informed by data use, the plants also provide visual collected from the field. Basking spot barriers for the frogs. temperatures, UV exposure and gradients, temperature and humidity Lids should be constructed of mesh as r e g i m e s s h o u l d m i m i c t h e glass will block UVB. environmental conditions at the collection site. Note that environmental Substrate must be provided for this data should be collected at different species as it is unknown whether or not times throughout the day and year. they like to dig. The provision of Suitable ambient room temperatures substrate will prevent the enclosure should vary between 23 and 27˚C from drying out. Be sure to incorporate a (night/day summer) and 20 and 25˚C drain in the tank construction. A layer of (night/day winter). gravel covered with a permeable matting should be laid underneath a substrate of A UV gradient should be provided in soil mixed with sand (ratio 2:1). The conjunction with a basking spot. Arcadia depth of substrate will depend on D3+ reptile lamp T5 12% UVB are ideal enclosure size and the size of the and a maximum UVI of 3 should be occupants, but must be at least 100 mm provided for the frogs at their basking deep. The larger the enclosure, the spots. Remember that mesh will filter deeper the substrate should be as the out some of the UVB so UV readings substrate will dry out quickly in large must be taken from below the mesh with enclosures. Substrate does not need to a solarmeter 6.5. be compacted for this species. Ensure that substrates are not contaminated Basking spots can be provided with with fertilisers or other substances that regular incandescent lamps. The may be toxic to amphibians. basking lamp should be positioned at one end of the enclosure and the Ensure the substrate has a humidity temperature on the substrate below the gradient by pouting some water (or basking spot should range between 30- spraying with water) more at the end of 32oC and not exceed 32oC. Ensure that all the enclosure with no basking light. lighting is inaccessible to the frogs as These are litter frogs and are well they will be burnt easily if they come into camouflaged, a layer of leaf litter should contact with lamps.These frogs can be provided for these frogs as they may often be found on dappled forest floor experience some level of stress without during the day (Pers. obs) and so the it. provision of UV lighting is essential.

Check the source of your water by testing it regularly for ammonia, nitrite, nitrate, pH and phosphates. Tap water should be left to stand for 24 hours before it is used in order for the chlorine to dissipate. Ensure the temperature of the water is at equilibrium with the room temperature. A plastic water dish should be provided and should measure 200 x 150 mm. Water depth should be 80 mm, and frogs should be able to immerse their drink Figure 6. Set up suitable for housing and breeding patch. An emergent rock should be C. curtipes (Zoological Society of London) placed in the water to allow the frogs

26 Ex situ Management of Amphibians easy access out of the water. f r o g s t h e y p r o b a b l y f e e d o n invertebrates. Food should be offered The enclosure should be lightly misted after it is dusted with an appropriate with aged tap water daily. dietary supplement, well fed and an appropriate size, no greater in size than Routine Husbandry the width of the frogs head. C. All animals should be visually inspected clinotarsus are nocturnal and should be daily. Water dishes must be cleaned out fed in the evening. and refilled daily as amphibians often excrete waste products in water. Water Adults should be fed every three to six quality cannot be judged by eye. Faeces days (depending on season and and uneaten food must be removed condition), juveniles up to six weeks of daily. The enclosure and all furniture age should be fed daily. must be thoroughly scrubbed (with a brush and hot water, no chemical cleaners or disinfectants) twice a week. Reproduction in Captivity Temperatures of the enclosure should be checked at different points of the Breeding Enclosure enclosure regularly to ensure that the Increasing the humidity may trigger correct thermoregulatory gradients are breeding in this species. These frogs maintained. UV readings must be taken should be cycled in a rain chamber with a every three months to check that there is sprinkler system for breeding (Fig. 7). a sufficient UV gradient and that the UV emitting lights are still emitting UV. A rain chamber with a floor space of 600 x 1000 mm would be suitable as a The substrate must be changed as soon minimum size for 2 males and 3 females. as it looks dirty or if it becomes saturated It is important that the sex ratio in the or unable to retain humidity. breeding enclosure is not biased towards males this may be stressful for the Diet in captivity females). No substrate is required in the The feeding habits of this frog are totally r a i n c h a m b e r . B a s e d o n t h e i r unknown, but like other similar sized morphology C.curtipes are probably poor swimmers so it is vital that there are some rocks or logs in the water that the frogs can easily leave the water should they choose to do so. A pump powered spray system or fountain would be ideal for breeding this species but a mature biological filter should be incorporated into the rain chamber to maintain water quality. Ideally a sump could be used (Appendix 1). The, lighting and temperature parameters should be maintained as above. Visual barriers such as palm fronds or plastic plants could be used to provide cover for the frogs in their breeding enclosure if they fail to spawn after a couple of weeks.

The frogs should be fed at least every other day whilst they are in the rain chamber but ensure that any dead Figure 7. A rain chamber suitable for breeding C. curtipes insects are removed daily as they will (Benjamin Tapley) 27 Ex situ Management of Amphibians pollute the water. Egg Care, Tadpole Husbandry & Development In closed systems regular water tests A description of the eggs of C. curtipes should be carried out (Temperature, pH, have not been described. When eggs are ammonia, nitrite and nitrate), the laid transfer them to a mature aquatic results will help you decide the system soon after laying so that they are frequency of water changes. pH should not damaged or exposed to the air by the not fluctuate, ammonia should measure frogs moving around in the rain less than 1 part per million (ppm), nitrite chamber. You may not wish to keep all less than 0.1ppm and nitrate less than the eggs if the clutch size is large as 10ppm. No more than 20% of the water rearing large numbers of tadpoles and should be changed as rapid shifts in providing food for all the metamoprhs water quality will be problematic for the will be a huge task. biological filtration process. As the tadpoles of this species are large Breeding Seasonality and form large shoals, tank size is There is no published report on the important. A tank measuring 1,000 x captive breeding of C. curtipes. In the 500 x 500 mm would be suitable for a wild, breeding aggregations of this maximum 250 tadpoles. Tanks should species can be found at the start of the be stocked with no more than 1 tadpole monsoon. per litre. Filtration is vital (see appendix one to inform your choice of filter). Aged Provision of Breeding Sites tap water can be used to rear tadpoles. Check the source of your water by testing A depth gradient should be provided for it regularly for ammonia, nitrite, nitrate, the frogs so that they can choose their hardness, pH and phosphates. Water egg laying site, this can be provided by parameters should be informed by field angling slates or paving slabs into the data but remember that not all breeding water (Fig.8). sites may have optimal water quality. Tap water should be left to stand for 24 hours before it is used in order for the chlorine to dissipate. Ensure the t e m p e r a t u r e o f t h e w a t e r i s a t equilibrium with the room temperature. Check the water in closed systems and c a r r y o u t r e g u l a r w a t e r t e s t s (temperature, pH, ammonia, nitrite and nitrate), the results will help you decide the frequency of water changes. pH should not fluctuate, ammonia should measure less than 1 part per million (ppm), nitrite less than 0.1ppm and nitrate less than 10ppm. No more than 20% of the water should be changed as rapid shifts in water quality will be problematic for the biological filtration and tadpoles. Ideally a sump should be used to increase the volume of water of your system and to house all the life support systems (filters etc). If using an Figure 8. A basic rain chamber used to successfully breed internal canister filter ensure that it is the Vietnamese bony headed frog (Ingerophrynus galeatus) in captivity. Note the sloped tile which provided the toads with not too strong as the tadpoles could be a depth gradient in the rain chamber (Benjamin Tapley). sucked into the inflow of the filter and

28 Ex situ Management of Amphibians may have difficulty feeding. Direct the they can easily drown. Emerging rocks outflow to the surface of the water for or aquatic plants are suitable. Once they oxygenation. emerge they are prone to drying out and easily die if the humidity is not high. When the eggs first hatch the tadpoles will not move and it is perfectly normal Rearing Metamorphs for them to sometimes rest on their side. Metamorphs can be housed in small If you look on the underside of tadpoles plastic containers. A 350 x 200 x 300 when they first hatch you should be able mm enclosure is suitable for up to 12 to see the yolk sac. The tadpole will not individuals (Fig. 9), these groups should start swimming or feeding until this ac be divided amongst larger enclosures as has been absorbed. Once the newly individuals grow. It is important that hatched tadpoles have dropped off the rearing tanks are relatively small so that egg mass it is important to remove the high ambient humidity can me remaining egg jelly as this will decay and maintained and that the juvenile frogs pollute the water. Substrate is not can find food easily. Covering the lid necessary but tadpoles like to hide. with a fine breathable cloth or fleece will D r i e d I n d i a n a l m o n d l e a v e s prevent the food escaping and ensure (Terminalia catappa) can be added, that humidity remains high. The cloth they also soften the water which would will block dome of the UV emitted from be beneficial in hard water areas. Partial the lights so you must experiment and (10-20 %) water changes should be e n s u r e t h a t t h e U V l i g h t a n d carried out regularly, the frequency of temperature gradients are still suitable. water changes must be governed by Compacted damp soil mixed with sand frequent water testing. would make a good substrate, and this must be replaced as necessary. It is Tadpoles should be fed daily, once they important that the substrate is start free swimming. They should be fed compacted when housing metamorphs on a variety of food. At Durrell ranid frog as the soil particles easily stick to the tadpoles were successfully reared on a skin of the froglets. A shallow water dish powdered tadpole food (components: must be provided (water depth 10 mm). ground tropical fish flake, grass pellet, It is essential that the water dish is easy trout pellets, tubifex, river shrimp, for the metamorphs to climb out of (a spirulina algae and cuttlefish bone). rock or small branch in the water dish Food should be available to the tadpoles facilitates access). When transferring at all times but it is important that large froglets from the tadpole tank make sure amounts of uneaten food do not pollute that they are placed in the water dish and the aquarium. Tadpole feeding regimes not on the substrate as they may not be must be modified, as the tadpoles grow quite ready to leave the water, they will they will eat more. choose when they are ready. Cover objects are extremely important Once the front limbs appear and the tail microhabitats for small frogs. Bark hides starts to reduce it is important that the and leaf litter should be added to the metamorphosing frolets are given the enclosure. The furnishings in the opportunity to emerge from the water as enclosure should not be too complex or it will be difficult for the young frogs to find food. The metamorphs can be raised at the same room temperature as the adults (23 - 27˚C), and must be misted with room temperature tap water twice daily. It is vital that metamorphs and juveniles are provided with UV at this critical growing stage (Arcadia D3+ Figure 9. Enclosure for rearing juvenile C. curtipes (Benjamin Tapley). 29 Ex situ Management of Amphibians reptile lamp T5 12% UVB), this must be Hiragond, N.C., Shanbhag, B.A., given in a gradient with a maximum UVI Saidapur, S.K. (2001): Description of the of 2 (Fig. 9). tadpole of a stream breeding frog, Rana curtipes. Journal of Herpetology. 35: Do not provide metamorphs with a 35oC 166-168 t basking spot as this will dry them out rapidly and will likely kill them. A lower IUCN, Conservation International, and temperature basking spot can be NatureServe. 2006. Global Amphibian provided with incandescent lamps Assessment.. Accessed on 09 October 2007. 1 The temperature on the branching below the basking spot should not Krishna, S.N., Krishna, S.B. (2005): exceed 28oC. Small tanks will rapidly Female courtship calls of the litter frog overheat so ensure that there is still a (Rana curtipes) in the tropical forests of cool area in the enclosure. Metamorphs Western Ghats, South India. Amphibia- should be fed daily until about six weeks Reptilia. 26: 431-435 of age with pin head crickets and (occasionally) Drosophila. All food 2 Krishna, S.N., Krishna, S.B. (2005): items should be dusted with appropriate Habitat of the Bicolor frog, Clinotarsus dietary supplements. After 8 weeks the curtipes, in the Western Ghats, South feeding interval can be gradually India. Herpetological. Review. 36:384- increased to once every 3 days. 387.

Health Stuart, S., Hoffmann, M., Chanson, J., Cox, N., Berridge, R., Ramani, P., and Problems Encountered in Young, B. (eds) (2008). Threatened Captivity Amphibians of the World. Lynx Non applicable. Edicions, IUCN, and Conservation International, Barcelona, Spain; Gland, Record keeping Switzerland; and Arlington, Virginia, USA. Things to include. Individual identification, weight, Tapley, B. & Purushotham. C.B (2011) feeding records, temperature records, Fighting behaviour in the Bicoloured UV records, water quality test results, frog Clinotarsus (Rana) curtipes Jerdon, h e a l t h r e c o r d s , b e h a v i o u r a l o r 1854. Herpetology Notes 4: 353-355. reproductive observations.

Bibliography Daniels, R, R, J. 2005. Amphibians on peninsular India. Universities press, Hyderabad. 288pp

30 Ex situ Management of Amphibians

6. Ex-situ Managemnt of Common tree frogs Polypedates species

Indonesia and the Philippines. It has been introduced to Japan and Papua New Guinea. It can be found from sea level up to 1, 500 m asl (IUCN et al., 2006).

Polypedates maculatus is restricted to including most of peninsular India, Sri Lanka, , and (IUCN et al., 2006).

Conservation Status and Threats Both species are listed as Least Concern by the IUCN Red List due to its extensive distribution and its tolerance of a wide range of habitat types(IUCN et al., 2006). It should be noted, however, that P. leucomystax represents a complex of cryptic species, and taxonomic revision

Figure 1. Top Polypedates leucomystax (Benjamin Tapley). of the complex is required (IUCN et al., Bottom Polypedates maculatus (Benjamin Tapley) 2006).

Introduction Habitat and Ecology Polypedates are very adaptable and Description occurs in a wide variety of habitats, Common tree frogs are slender including beach vegetation, various rhacophorid tree frogs with a pointed human habitats, natural edge habitats, snouts and prominent eyes (Fig.1). P. and closed subtropical or tropical forest l e u c o m y s t a x i s p o l y m o r p h i c , (IUCN et al., 2006). colouration and patterning are highly variable, even amongst wild specimens Sexing Individuals collected in the same area (Church, Polypedates are sexually dimorphic 1963). The end of each toe terminates in with females reaching maximum SVLs a large disc, enabling this species to u p t o 5 0 % l a r g e r t h a n m a l e s . climb well. Females of P. leucomstax Reproductively active males have may grow to 80 mm snout vent length nuptial pads at the base of the first (SVL) and males to 60 mm SVL. Males of finger. P. maculatus can measure 34-57mm SVL and females up to 89mm SVL Reproduction and Larval (Amphibia web, Daniels, 2005) Development I n d r i e r p a r t s o f i t s r a n g e , P . Distribution leucomystax, breeds during the wet Polypedates leucomystax is a widely season. In wetter areas of its range the distributed species, occurring in species reportedly breeds year round. northeast India, Bangladesh, Nepal, Polypedates maculatus breeds during western Yunnan (China) and most of the monsoon rains (Amphibiaweb) . The species is Males congregate around slow moving also found throughout much of or still water bodies. They sit in elevated positions and call to females to attract 311 Ex situ Management of Amphibians them. The call is a nasal quack. Females Identifying Individuals of P. leucomystax respond to the males Adult specimens of both species are not vocalisations by tapping their toes, the individually identifiable as specimens males then move closer to the females do not possess characteristic markings. (Narins, 1995). Adults could be marked with passive integrated transponders (PIT tags). Pit Amplexus and Nest Production tags should be inserted subcutaneously The breeding biology of P. leucomystax on the dorsum. Juveniles could be is well documented. Amplexus (mating marked with different coloured visible position) is axillary in these species. implant elastomer (VIE) but this may Once in amplexus the female deposits up not be necessary for rearing purposes to 500 eggs (Average egg diameter = 1.85 (Fig. 2). mm) on a surface overhanging water (Duellman & Trueb, 1994). The male fertilises the eggs and both sexes secrete mucus. The male uses his hind legs to whisk this up into a foam. This process can take 12 hours. The resulting ball of foam measures approximately 100mm across (Yorke, 1983). Mass nesting has been observed in the wild, the resulting egg mass measured a metre across Figure 2. P. leucomystax metamorph marked with VIE (Gaulke & Cadiz, 2002). The surface of (Benjamin Tapley) the nest hardens after a couple of hours (Staniszewski, 1995). In captivity Housing adults females can produce more than one nest At Durrell P. leucomystax was housed per breeding season (pers. obs.). and bred in two different types of enclosure. The first enclosure was a 500 The breeding biology of P. maculatus is x 500 x 900 mm glass vivarium with a not been studied in detail but the secure mesh lid. A partition divided the reproductive strategy is identical to P. floor of the vivarium into a water area leucomystax. (70 mm deep) and a land area. Coarse gravel was used as the substrate for the Longevity and Age at Sexual land area. This would be the minimum Maturity enclosure size for 2 males and 2 females. Lifespan and age at sexual maturity in The second enclosure was a converted the wild are unknown. The maximum plastic storage unit. It measured 600 x lifespan in captivity is also unknown; to 800 x 600 mm, and had a wire mesh lid. date captive P. leucomystax have been A 250 x 150 x 100 mm water dish was recorded to live 6 to 8 years. Sexual provided, this would be suitable for 6 maturity can be reached at one of age in adult individuals. No substrate was captivity. provided in this enclosure (Fig. 3).

Captive Management Branching is essential when keeping arboreal tree frogs. Multiple bamboo Introduction lengths (at least 30 mm diameter) are P. leucomystax is commonly kept by ideal. The ends of the bamboo used for zoos and private individuals, and has branching should be sealed to prevent been bred in captivity for decades. live food hiding. 150 mm lengths of PVC P o l y p e d a t e s m a c u l a t u s i s n o t pipe or hollowed bamboo (70 mm established in captivity. diameter) can be attached to the bamboo branching to provide hiding places for the frogs. Branches should be positioned

321 Ex situ Management of Amphibians at different angles within the enclosure (night/day summer) and 20 and 25˚C with some under the basking spot and (night/day winter). UV light and others facing away, this will allow the frogs to regulate their body A UV gradient should be provided in temperature, rate of water loss through conjunction with a basking spot. Arcadia their permeable skin and UV exposure. D3+ reptile lamp T5 12% UVB are ideal Live plants can be added as they create and a maximum UVI of 3 should be humid pockets which the animals may provided for the frogs at their basking use as perching sites, the plants also spots. Remember that mesh will filter provide visual barriers for the frogs. out some of the UVB so UV readings must be taken from below the mesh with Lids should be constructed of mesh as a solarmeter 6.5. glass will block UVB. Basking spots can be provided with Substrate can be provided for this regular incandescent lamps. The species although it is not essential. The basking lamp should be positioned at provision of substrate will prevent the one end of the enclosure and the enclosure from drying out. Be sure to temperature on the branching below the incorporate a drain in the tank basking spot should range between 30- construction. A layer of gravel covered 35oC and not exceed 35oC. Ensure that all with a permeable matting should be laid lighting is inaccessible to the frogs as underneath a substrate of soil mixed they will be burnt easily if they come into with sand (ratio 2:1). The depth of contact with lamps. substrate will depend on enclosure size. The larger the enclosure, the deeper the Check the source of your water by testing substrate should be as the substrate will it regularly for ammonia, nitrite, nitrate, dry out quickly in large enclosures. pH and phosphates. Tap water should be Substrate should be compacted to left to stand for 24 hours before it is used prevent soil sticking to the skin of the in order for the chlorine to dissipate. frog. Ensure that substrates are not Ensure the temperature of the water is at contaminated with fertilisers or other equilibrium with the room temperature. substances that may be toxic to A plastic water dish should be provided amphibians. and should measure 200 x 150 mm. Water depth should be 80 mm, and frogs Ensure the substrate has a humidity should be able to immerse their drink gradient by pouting some water (or patch. An emergent rock should be spraying with water) more at the end of placed in the water to allow the frogs the enclosure with no basking light. easy access out of the water.

Temperature, Humidity and The enclosure should be lightly misted Lighting with aged tap water daily. Husbandry should be informed by data collected from the field. Basking spot Routine Husbandry temperatures, UV exposure and All animals should be visually inspected gradients, temperature and humidity daily. Water dishes must be cleaned out r e g i m e s s h o u l d m i m i c t h e and refilled daily as amphibians often environmental conditions at the excrete waste products in water. Water collection site. Note that environmental quality cannot be judged by eye. Faeces data should be collected at different and uneaten food must be removed times throughout the day and year. At daily. The enclosure and all furniture Durrell P. leucomystax were maintained must be thoroughly scrubbed (with a in a room with ambient temperatures brush and hot water, no chemical which varied between 23 and 27˚C cleaners or disinfectants) twice a week.

331 Ex situ Management of Amphibians

Temperatures of the enclosure should be age should be fed daily. checked at different points of the enclosure regularly to ensure that the Reproduction in Captivity correct thermoregulatory gradients are maintained. UV readings must be taken Breeding Enclosure every three months to check that there is Increasing the humidity can trigger a sufficient UV gradient and that the UV breeding in Polypedates. Increasing the emitting lights are still emitting UV. humidity can be achieved by increasing the amount that the frogs are sprayed on The substrate must be changed as soon daily basis. Ideally these frogs should be as it looks dirty or if it becomes saturated cycled in a rain chamber with a sprinkler or unable to retain humidity. system (Fig. 4).

A rain chamber measuring 500 x 500 x 900 mm would be suitable as a minimum size for 2 males and 2 females. It is important that the sex ratio in the breeding enclosure is not biased towards males as this can be stressful for the females. No substrate is required in the rain chamber A pump powered spray system would be ideal for breeding this species but a mature biological filter should be incorporated into the rain chamber to maintain water quality.

Figure 3. One set up for housing and breeding P. leucomystax (Benjamin Tapley)

Diet in captivity At Durrell P. leucomystax were fed on live invertebrates, predominantly crickets, cockroaches and locusts. Juvenile animals were fed on live pin head crickets and Drosophila hydei. All food items were dusted with Nutrobal® (vitamin and mineral supplement) immediately prior to being fed out. Food should be well fed and an appropriate size, no greater in size than the width of the frogs head. Polypedates are nocturnal and should be fed in the evening.

Adults should be fed every three to six days (depending on season and condition), juveniles up to six weeks of Figure 4. A rain chamber suitable for breeding Polyedates (Andrew Tillson-Willis) 341 Ex situ Management of Amphibians

Ideally a sump could be used (Appendix directly below the nest for tadpoles to 1). The branching, lighting and drop down into (Fig. 5). The foam nest temperature parameters should be must be lightly sprayed with tap water maintained as above. daily.

The frogs should be fed at least every other day whilst they are in the rain chamber but ensure that any dead insects are removed daily as they will pollute the water.

In closed systems regular water tests should be carried out (Temperature, pH, ammonia, nitrite and nitrate), the results will help you decide the frequency of water changes. pH should not fluctuate, ammonia should measure Figure 5. Foam nest of P. leucomystax. (Benjamin Tapley) less than 1 part per million (ppm), nitrite less than 0.1ppm and nitrate less than Tadpole Husbandry and 10ppm. No more than 20% of the water Development should be changed as rapid shifts in After hatching, tadpoles can be water quality will be problematic for the transferred to a glass tank with a spoon biological filtration process. (Fig. 6), they are extremely delicate at this stage so they should be moved with Breeding Seasonality utmost caution. When they first hatch In captivity P. leucomystax breed during they will not move and it is perfectly the warmer months. At Durrell this was normal for them to sometimes rest on from April through to October. their side. If you look on the underside of Polypedates maculatus have not been tadpoles when they first hatch you bred in captivity. should be able to see the yolk sac. The tadpole will not start swimming or Provision of Breeding Sites feeding until this ac has been absorbed. Adults need branching over-hanging The size of the tank for tadpoles is not water on which to make their foam nest. important, the stocking density is. Tanks They may also nest in leaves or attach should be stocked with no more than 5 their nests to the side of the enclosure or tadpoles per litre. Filtration is vital (see water dish. appendix one to inform your choice of filter). Aged tap water can be used to rear Care of Foam Nest tadpoles. Check the source of your water It may take several weeks in the rain by testing it regularly for ammonia, chamber for breeding to occur. Once nitrite, nitrate, hardness, pH and hardened, the nest can be removed from phosphates. Water parameters should the enclosure to facilitate collection of be informed by field data but remember tadpoles. Nests can be gently removed that not all breeding sites may have from the deposition site and stuck (the optimal water quality, especially in underside of the nest remains sticky) to urbanised environments where these the side of a large plastic container. This frogs can often be found. Tap water container must then raised at a slight should be left to stand for 24 hours angle and the bottom of the container before it is used in order for the chlorine filled with water. This created a deeper to dissipate. Ensure the temperature of area of water (approximately 15 mm) at the water is at equilibrium with the room the end of the container furthest from temperature. Check the water in closed the nest. It is essential that there is water systems and carry out regular water tests 351 Ex situ Management of Amphibians

(temperature, pH, ammonia, nitrite and amounts of uneaten food do not pollute nitrate), the results will help you decide the aquarium. Tadpole feeding regimes the frequency of water changes. pH must be modified, as the tadpoles grow should not fluctuate, ammonia should they will eat more. measure less than 1 part per million (ppm), nitrite less than 0.1ppm and Once the front limbs appear and the tail nitrate less than 10ppm. No more than starts to reduce it is important that the 20% of the water should be changed as metamorphosing froglets are given the rapid shifts in water quality will be opportunity to emerge from the water as problematic for the biological filtration they can easily drown. Emerging rocks and tadpoles. Ideally a sump should be or aquatic plants are suitable. Once they used to increase the volume of water of emerge they are prone to drying out and your system and to house all the life easily die if the humidity is not high. support systems (filters etc). If using an Rearing Metamorphs Metamorphs can be housed in small plastic containers. A 350 x 200 x 300 mm enclosure is suitable for up to 12 individuals, these groups should be divided amongst larger enclosures as individuals grow. It is important that rearing tanks are relatively small so that Figure 6. Polypedats maculatus tadpole (Sandeep Varma) high ambient humidity can me internal canister filter ensure that it is maintained and that the juvenile frogs not too strong as the tadpoles could be can find food easily. Covering the lid sucked into the inflow of the filter and with a fine breathable cloth or fleece will may have difficulty feeding. Direct the prevent the food escaping and ensure outflow to the surface of the water for that humidity remains high. The cloth oxygenation. will block dome of the UV emitted from the lights so you must experiment and Substrate is not necessary but tadpoles e n s u r e t h a t t h e U V l i g h t a n d like to hide. Dried Indian almond leaves temperature gradients are still suitable. (Terminalia catappa) can be added, Damp soil mixed with sand would make they also soften the water which would a good substrate, and this must be be beneficial in hard water areas. Partial replaced as necessary. A shallow water (10-20 %) water changes should be dish must be provided (water depth 10 carried out regularly, the frequency of mm). It is essential that the water dish is water changes must be governed by easy for the metamorphs to climb out of frequent water testing. Polypedates (a rock or small branch in the water dish leucomystax tadpoles metamorphosed facilitates access). When transferring after approximately 60 - 90 days at 23 – froglets from the tadpole tank make sure 26oC. that they are placed in the water dish and not on the substrate as they may not be Tadpoles should be fed daily once they quite ready to leave the water, they will start swimming freely. They should be choose when they are ready. Live plants fed on a variety of food. At Durrell should be included in the juvenile tadpoles were successfully reared on a enclosures. The metamorphs can be powdered tadpole food (components: raised at the same room temperature as ground tropical fish flake, grass pellet, the adults (23 - 27˚C), and must be trout pellets, tubifex, river shrimp, misted with room temperature tap water spirulina algae and cuttlefish bone). twice daily. It is vital that metamorphs Food should be available to the tadpoles and juveniles are provided with UV at at all times but it is important that large this critical growing stage (Arcadia D3+ 361 Ex situ Management of Amphibians reptile lamp T5 12% UVB), this must be five minutes daily for 11 days. There was given in a gradient with a maximum UVI z e r o m o r t a l i t y f o r s p e c i m e n s of 2. Do not provide metamorphs with a undergoing this treatment. 35oC t basking spot as this will dry them out rapidly and will likely kill them. A Specimens were treated for parasites lower temperature basking spot can be using Levamisole. Specimens were provided with incandescent lamps bathed in baths (60mg/kg) for 30 positioned at one end of the enclosure. minutes once a week for three weeks. No The temperature on the branching mortality was observed for specimens below the basking spot should not undergoing this treatment. exceed 28oC. Small tanks will rapidly overheat so ensure that there is still a Record keeping cool area in the enclosure. Metamorphs should be fed daily until about six weeks Things to include. of age with pin head crickets and Individual identification, weight, (occasionally) Drosophila. All food feeding records, temperature records, items should be dusted with appropriate UV records, water quality test results, dietary supplements. After 8 weeks the h e a l t h r e c o r d s , b e h a v i o u r a l o r feeding interval can be gradually reproductive observations. increased to once every 3 days. Bibliography Health Church, G. .1963. The variation in dorsal pattern in Rhacophorus leucomystax. Problems Encountered in Copeia 1963: 400-405

Captivity Christensen-Dalsgaard, J., Ludwig, T. A. & Narins, P. M. 2002. Complex vocal communication in the Southeast Asian f r o g P o l y p e d a t e s l e u c o m y s t a x . Bioacoustics. 13: 21-25.

Daniels, R, R, J. 2005. Amphibians on peninsular India. Universities press, Hyderabad. 288pp

Duellman, W. E. & Trueb, L. 1994. Figure 7. Rostral abrasion in a female P. leucomystax (Benjamin Tapley) Biology of Amphibians. John Hopkins This species appears susceptible to University Press, Baltimore. 672pp rostral abrasions (Fig. 7). The provision of adequate space is important in IUCN, Conservation International, and minimising / avoiding the occurrence of NatureServe. 2006. Global Amphibian rostral lesions. Assessment.. Accessed on 09 October 2007. Routine Veterinary Procedures At Durrell Wildlife Conservation Narrins, P.M. 1995. Comparative Trust aspects of interactive communication Pp Specimens have been given intra- 363-372 In Active hearing, Ed A. Flock, muscular injections of antibiotics (e.g. D. Ottossen, and M. Ulfendahl. Elsevier Baytril, 10 mg/kg). Science, London

All imported specimens were treated Staniszewski, M. (1995) Amphibians in against chytridiomycosis by bathing Captivity. T.F.H Publications INC, U.S.A them in Itroconoazole solution (1:99) for 544 pp.

371 Ex situ Management of Amphibians

7. Ex-situ Management of Gliding frog Rhacophorus species

Conservation Status and Threats Both species are listed as Least Concern by the IUCN Red List due to its extensive distribution and its tolerance of a wide range of habitat types(IUCN et al., 2006)..

Habitat and Ecology Rahcophorus malabaricus inhabits of Figure 1. Top Rhacophorus malabaricus (Benjamin Tapley). tropical moist evergreen forest, deciduous forest, secondary forest and Introduction coffee plantations. Rhacophorus maximus inhabits lowland to sub Description montane moist evergreen forest (IUCN Gliding frogs are rhacophorid tree frogs et al., 2006). with pointed snouts, prominent eyes and extensive webbing between the Sexing Individuals fingers and toes (Fig.1). These frogs are Rhacophorus are sexually dimorphic, leaf green in colour. Rhacophorus females being much larger than males. malabaricus is green in colouration with yellow / orange webbing on the feet Reproduction and Larval (Danels, 2005). Rhacophorus maximus Development is the largest tree frog in India and can be Rhacophorus malabaricus breed after distuinguished from other Rhacophorid the onset of the monsoon in June and frogs in the region by thr brown July (Daniels, 2005). Eggs hatch after 4- reticulations on the flanks and webbing 5 days and larval development is report and a white stripe on the lower jaw to take 68 days (Daniels, 2005). (Ahmed et al., 2009). The end of each to Rhacophorus maximus breed from in Rhacophorus species terminate in a February to April in Meghalya (Ahmed large disc, enabling these frogs to climb et al., 2009). well. Females of R. malabaricus may grow to 92 mm snout vent length (SVL) Males congregate around slow moving (Pers. Obs). Rhacophorus maximus or still water bodies. They sit in elevated may attain a SVL of 93mm ((Ahmed et positions and call to females to attract al., 2009) them.

Distribution Amplexus and Nest Production Rhacophorus malabaricus is restricted See fig. 2. The breeding biology of R. to the Western Ghats of India, it has an malabaricus is well documented altitudinal range of 300 – 1,200 masl. (Kadadevaru & Kanamadi, 2000). (IUCN et al., 2006) Amplexus (mating position) is axillary in this species. Once in amplexus the Rhacophorus maximus is known from female deposits up to 192 eggs on a eastern Nepal, northern Bangladesh, surface overhanging or next to water North East India, northern , (Kadadevaru & Kanamadi, 2000). The western and southern China male fertilises the eggs (average and has been recorded from 500- diameter 2.62 mm) and both sexes 2,000m asl. (IUCN et al., 2006). secrete mucus. The male uses his hind legs to whisk this up into a foam which is 381 Ex situ Management of Amphibians wrapped in leaves by the hind limbs of Adults could be marked with passive the frogs (Kadadevaru & Kanamadi, integrated transponders (PIT tags). Pit 2000). tags should be inserted subcutaneously on the dorsum. Juveniles could be marked with different coloured visible implant elastomer (VIE) but this may not be necessary for rearing purposes

Housing adults M i n i m u m e n c l o s u r e s i z e f o r 5 individuals (2 males 5 females) 800 x 800 x 1500 mm, a mesh lid must be provided. Due to the jumping nature of these frogs, front opening enclosures are recommended. A 250 x 150 x 100 mm water dish must be provided (Fig 3.).

Branching is essential when keeping arboreal tree frogs. Multiple bamboo lengths (at least 30 mm diameter) are ideal. The ends of the bamboo used for branching should be sealed to prevent live food hiding. 150 mm lengths of PVC pipe or hollowed bamboo (70 mm diameter) can be attached to the bamboo branching to provide hiding places for the frogs. Branches should be positioned at different angles within the enclosure with some under the basking spot and Figure 2. Foam nest of R.. malabaricus (Benjamin Tapley). UV light and others facing away, this will The breeding biology of R. maximusis allow the frogs to regulate their body not been studied in the field detail but temperature, rate of water loss through the reproductive strategy is identical to their permeable skin and UV exposure. R. malabaricus. Live plants can be added as they create humid pockets which the animals may Longevity and Age at Sexual use as perching sites, the plants also Maturity provide visual barriers for the frogs. Lifespan and age at sexual maturity in the wild are unknown. Lids should be constructed of mesh as glass will block UVB. Captive Management Substrate can be provided for this Introduction species although it is not essential. The Rahcophorus maximus is kept by zoos provision of substrate will prevent the and private individuals, and has been enclosure from drying out. Be sure to bred in captivity. Rahcophorus incorporate a drain in the tank malabaricus is not established in construction. A layer of gravel covered captivity. with a permeable matting should be laid underneath a substrate of soil mixed Identifying Individuals with sand (ratio 2:1). The depth of Adult specimens of both species are not substrate will depend on enclosure size. individually identifiable as specimens The larger the enclosure, the deeper the do not possess characteristic markings. substrate should be as the substrate will

391 Ex situ Management of Amphibians dry out quickly in large enclosures. particular as this species has been Substrate should be compacted to observed sitting out during the day in prevent soil sticking to the skin of the direct sun (Fig 4.). frog. Ensure that substrates are not contaminated with fertilisers or other Basking spots can be provided with substances that may be toxic to regular incandescent lamps. The amphibians. basking lamp should be positioned at one end of the enclosure and the Ensure the substrate has a humidity temperature on the branching below the gradient by pouting some water (or basking spot should range between 30- spraying with water) more at the end of 32oC and not exceed 32oC. Ensure that all the enclosure with no basking light. lighting is inaccessible to the frogs as they will be burnt easily if they come into Temperature, Humidity and contact with lamps. During the winter Lighting the basking spot for R. maximus should Husbandry should be informed by data not exceed 25oC. collected from the field. Basking spot temperatures, UV exposure and Check the source of your water by testing gradients, temperature and humidity it regularly for ammonia, nitrite, nitrate, r e g i m e s s h o u l d m i m i c t h e pH and phosphates. Tap water should be environmental conditions at the left to stand for 24 hours before it is used collection site. Note that environmental in order for the chlorine to dissipate. data should be collected at different Ensure the temperature of the water is at times throughout the day and year equilibrium with the room temperature. especially for these species which may A plastic water dish should be provided vary in the preference from site to site and should measure 200 x 150 mm. due to the range of altitudes in which Water depth should be 80 mm, and frogs they are known to occur. should be able to immerse their drink patch. An emergent rock should be As a guideline for R. malabaricus should placed in the water to allow the frogs be housed in a room with ambient easy access out of the water. temperatures which vary between 23 and 27˚C (night/day summer) and 20 The enclosure should be lightly misted and 25˚C (night/day winter). R. with aged tap water daily. maximus should be kept cooler 18 and 24˚C (night/day summer) and 12 and Routine Husbandry 16˚C (night/day winter). A UV gradient All animals should be visually inspected should be provided in conjunction with a daily. Water dishes must be cleaned out basking spot. Arcadia D3+ reptile lamp and refilled daily as amphibians often T5 12% UVB are ideal and a maximum excrete waste products in water. Water UVI of 3 should be provided for the frogs quality cannot be judged by eye. Faeces at their basking spots. Remember that and uneaten food must be removed mesh will filter out some of the UVB so daily. The enclosure and all furniture UV readings must be taken from below must be thoroughly scrubbed (with a the mesh with a solarmeter 6.5. UV is brush and hot water, no chemical essential for R. malabaricus in cleaners or disinfectants) twice a week. Temperatures of the enclosure should be checked at different points of the enclosure regularly to ensure that the correct thermoregulatory gradients are maintained. UV readings must be taken every three months to check that there is a sufficient UV gradient and that the UV Figure 3. Diurnal resting site of Rhacphorus malabaricus (Benjamin Tapley) 401 Ex situ Management of Amphibians emitting lights are still emitting UV. chamber with a sprinkler system (Fig. 5). A rain chamber measuring 800 x 800 x The substrate must be changed as soon 1500 mm would be suitable as a as it looks dirty or if it becomes saturated minimum size for 2 males and 3 females. or unable to retain humidity. It is important that the sex ratio in the breeding enclosure is not biased towards males as this can be stressful for the females. No substrate is required in the rain chamber A pump powered spray system would be ideal for breeding this species but a mature biological filter should be incorporated into the rain chamber to maintain water quality. Ideally a sump could be used (Appendix 1). The branching, lighting and temperature parameters should be maintained as above.

Figure 4. A basic set up for housing Rahcophorus (Benjamin Tapley)

Diet in captivity Rhacophorus probably feed on large invertebrates and potentially small vertebrates. The feeding habits of R. malabaricus and R. maximus have not been studied in the field.

Adults should be fed every three to six days (depending on season and condition), juveniles up to six weeks of age should be fed daily.

Food should be offered after it is dusted with an appropriate dietary supplement, well fed and an appropriate size, no greater in size than the width of the frogs Figure 5. A rain chamber suitable for breeding head. Rhacophorus are nocturnal and Rhacophorus (Andrew Tillson-Willis) should be fed in the evening. The frogs should be fed at least every Reproduction in Captivity other day whilst they are in the rain chamber but ensure that any dead Breeding Enclosure insects are removed daily as they will Increasing the humidity can trigger pollute the water. breeding in Rhacophorus. These frogs should be cycled for breeding in a rain In closed systems regular water tests

411 Ex situ Management of Amphibians should be carried out (Temperature, pH, Tadpole Husbandry and ammonia, nitrite and nitrate), the Development results will help you decide the After hatching, tadpoles can be frequency of water changes. pH should transferred to a glass tank with a spoon. not fluctuate, ammonia should measure Tadpoles are extremely delicate at this less than 1 part per million (ppm), nitrite stage so they should be moved with less than 0.1ppm and nitrate less than utmost caution. When they first hatch 10ppm. No more than 20% of the water they will not move and it is perfectly should be changed as rapid shifts in normal for them to sometimes rest on water quality will be problematic for the their side. If you look on the underside of biological filtration process. tadpoles when they first hatch you should be able to see the yolk sac. The Breeding Seasonality tadpole will not start swimming or Rhacophorus malabaricus breed at the feeding until this ac has been absorbed. onset of the Monsson in June and July The size of the tank for tadpoles is not (Daniels, 2005). In Megahalya R. important, the stocking density is. Tanks maximus breed from February to April should be stocked with no more than 5 (Ahmed et al., 2009). tadpoles per litre. Filtration is vital (see appendix one to inform your choice of Provision of Breeding Sites filter). Aged tap water can be used to rear Adults need broad laves over-hanging tadpoles. Check the source of your water water on which to make their foam nest. by testing it regularly for ammonia, They may also nest in leaves or attach nitrite, nitrate, hardness, pH and their nests to the side of the enclosure or phosphates. Water parameters should water dish. be informed by field data but remember that not all breeding sites may have Care of Foam Nest optimal water quality, especially in It may take several weeks in the rain urbanised environments where these chamber for breeding to occur. Once frogs can often be found. Tap water hardened, the nest can be removed from should be left to stand for 24 hours the enclosure to facilitate collection of before it is used in order for the chlorine tadpoles. Nests can be gently removed to dissipate. Ensure the temperature of from the deposition site and stuck (the the water is at equilibrium with the room underside of the nest remains sticky) to temperature. Check the water in closed the side of a large plastic container. This systems and carry out regular water tests container must then raised at a slight (temperature, pH, ammonia, nitrite and angle and the bottom of the container nitrate), the results will help you decide filled with water. This created a deeper the frequency of water changes. pH area of water (approximately 15 mm) at should not fluctuate, ammonia should the end of the container furthest from measure less than 1 part per million the nest. It is essential that there is water (ppm), nitrite less than 0.1ppm and directly below the nest for tadpoles to nitrate less than 10ppm. No more than drop down into (Fig. 6). The foam nest 20% of the water should be changed as must be lightly sprayed with tap water rapid shifts in water quality will be daily. problematic for the biological filtration and tadpoles. Ideally a sump should be used to increase the volume of water of your system and to house all the life support systems (filters etc). If using an internal canister filter ensure that it is not too strong as the tadpoles could be sucked into the inflow of the filter and may have difficulty feeding. Direct the Figure 6. Foam nest of P. leucomystax. (Benjamin Tapley) 421 Ex situ Management of Amphibians outflow to the surface of the water for e n s u r e t h a t t h e U V l i g h t a n d oxygenation. temperature gradients are still suitable. Damp soil mixed with sand would make Substrate is not necessary but tadpoles a good substrate, and this must be like to hide. Dried Indian almond leaves replaced as necessary. A shallow water (Terminalia catappa) can be added, they dish must be provided (water depth 10 also soften the water which would be mm). It is essential that the water dish is beneficial in hard water areas. Partial easy for the metamorphs to climb out of (10-20 %) water changes should be (a rock or small branch in the water dish carried out regularly, the frequency of facilitates access). When transferring water changes must be governed by froglets from the tadpole tank make sure frequent water testing. that they are placed in the water dish and not on the substrate as they may not be Tadpoles should be fed daily once they quite ready to leave the water, they will start swimming freely. They should be choose when they are ready. Live plants fed on a variety of food. At Durrell should be included in the juvenile rhacophorid tadpoles were successfully enclosures. The metamorphs can be reared on a powdered tadpole food raised at the same room temperature as (components: ground tropical fish flake, the adults (23 - 27˚C), and must be grass pellet, trout pellets, tubifex, river misted with room temperature tap water shrimp, spirulina algae and cuttlefish twice daily. It is vital that metamorphs bone). Food should be available to the and juveniles are provided with UV at tadpoles at all times but it is important this critical growing stage (Arcadia D3+ that large amounts of uneaten food do reptile lamp T5 12% UVB), this must be not pollute the aquarium. Tadpole given in a gradient with a maximum UVI feeding regimes must be modified, as the of 2. Do not provide metamorphs with a tadpoles grow they will eat more. 35oC t basking spot as this will dry them out rapidly and will likely kill them. A Once the front limbs appear and the tail lower temperature basking spot can be starts to reduce it is important that the provided with incandescent lamps metamorphosing froglets are given the positioned at one end of the enclosure. opportunity to emerge from the water as The temperature on the branching they can easily drown. Emerging rocks below the basking spot should not or aquatic plants are suitable. Once they exceed 28oC. Small tanks will rapidly emerge they are prone to drying out and overheat so ensure that there is still a easily die if the humidity is not high. cool area in the enclosure. Metamorphs should be fed daily until about six weeks Rearing Metamorphs of age with pin head crickets and Metamorphs can be housed in small (occasionally) Drosophila. All food plastic containers. A 350 x 200 x 300 items should be dusted with appropriate mm enclosure is suitable for up to 12 dietary supplements. After 8 weeks the individuals, these groups should be feeding interval can be gradually divided amongst larger enclosures as increased to once every 3 days. individuals grow. It is important that rearing tanks are relatively small so that Health high ambient humidity can me maintained and that the juvenile frogs Problems Encountered in can find food easily. Covering the lid Captivity with a fine breathable cloth or fleece will Being a large gliding frog there is prevent the food escaping and ensure potential for rostral abrasions due to that humidity remains high. The cloth collision with the enclosure sides and or will block dome of the UV emitted from furnishings. the lights so you must experiment and

431 Ex situ Management of Amphibians

Record keeping Duellman, W. E. & Trueb, L. 1994. Biology of Amphibians. John Hopkins Things to include. University Press, Baltimore. 672pp Individual identification, weight, feeding records, temperature records, IUCN, Conservation International, and UV records, water quality test results, NatureServe. 2006. Global Amphibian h e a l t h r e c o r d s , b e h a v i o u r a l o r Assessment.. Accessed on 09 October 2007.

Bibliography Kadadevaru, G.G. & Kanamadi, K.D. 2000. Courtship and nesting behaviour Ahmed, F.M., Das, A & Dutta, S.K o f t h e M a l a b a r g l i d i n g f r o g , (2009) Amphibians and reptiles of Rhacophorus malabaricus (Jerdon, North East India. Aaranyak, Guwahati. 1870). Current Science 79 (3): 377-380. 168pp Mathew, R. & Sen, N. (2010) Pictorial Daniels, R, R, J. 2005. Amphibians on guide to amphibians of North East India. peninsular India. Universities press, Zoological Survey of India, Kolkata. Hyderabad. 288pp 144pp.

441 Ex situ Management of Amphibians

8. Ex-situ Management of Salamander Tylototriton verrucosus

and Kästle, 2002).

Conservation Status and Threats It has not been evaluated so far and information on their population trend assessment is scanty. In India, there is only one study which concludes that despite the high relative abundance of salamander in the area the habitats are

Figure 1. Top Tylototriton verrocosus (Shruti Sengupta). disappearing at a rapid rate and recommended raising the IUCN status Introduction of the species to “Critically Endangered” because of a reduction in the number of Description populations higher than 40% in 4 years It is amedium size salamander with SVL (Seglie, 2002). According to local ranging in male 87.70 - 63.10 mm, and people, sharing the same habitat for 97.00 - 66.00 mm in females, bony their day to day activities, the population ridges are present on head. Head is of T. verrucosus is probably declining, usually broad and flat, snout is blunt, but there is no data that could support moderate size eye with granular upper this notion at present (Vasudevan et al. eyelid. The two lines of dorsolateral bony 2014). ridges on head are distinct and are widely separated. The flanks have Habitat and Ecology roughly 16 pairs of longitudinal lines of The habitats are characterized as the knob-like dorsal warts that are distinct. subtropical hill forest and area is Skin ventrally and tail finely granular. dominated by scattered vegetation, for Tail is compressed laterally, with a well- example, Schima wallichii, Castonopsis developed fin fold. It is light brown indica, Casttonopsis tribuloides, Albizzi colour with darker warts in life. This sp., Sauraria nepalensis, Rubus species can produce loud vocalization e l l i p t i c u s a n d E u p a t o r i u m which has not been recorded so far in adenophorum. It occurs in Darjeeling other Tylototriton species hills from 1350-2000 m asl in streams and ponds that are surrounded by Distribution forests, tea plantations and villages Anders et al (1998) reported the (Deuti and Hegde 2007). Adults are seen distribution of the new species in five in water bodies only during the breeding places in the Illam district Nepal with season that spans from late April to early elevation ranging from 1100 m to October. During other months they are 2120m. The distribution of the species is terrestrial or semi-fossorial, hide under currently known only from the Illam rocks, logs and leaf and aestivate or district of Nepal and in Darjeeling hibernate for four to five months district, India (Nag and Vasudevan (Kuzmin et al. 1994). 2014). The salamanders are more terrestrial in non-breeding seasons (from October to February) and found hiding under the logs, bushes and stones and come to the breeding ponds in early March or April soon after heavy monsoonal showers (also see Schleich Figure 2. Breeding habitat of T. verrucosus (Shruti Sengupta). 451 Ex situ Management of Amphibians

Sexing Individuals of the existing facility at The Padmaja Naidu Himalayan Zoological Park In body length, tail length and weight (PNHZP), Darjeeling. The inputs from females were larger than males in males. field data on microhabitats used by the There was no distinct colour variation species in the breeding pools and between male and female. The dorsal proximate environmental cue required colouration varied from dark brown to for initiation of breeding should be taken light brown. Tail height is larger in into consideration in developing the females than in males. The vent length is design. Emphasis should be made on longer in male compared to female. housing animals on exhibit and off exhibit in the design. Facilities for water Reproduction and Larval treatment maintenance flow of water Development and humidity levels in the enclosures Females attain sexual maturity by the need to be incorporated in the design. age of 3-4 years (Kuzmin et al. 1994). During the breeding season that lasts up Identifying Individuals to end of monsoon; male salamanders Adult specimens of both species are not become brighter in colour, while in individually identifiable as specimens females the cloaca becomes orange- do not possess characteristic markings. colored. Clutch sizes vary from 26 to 60 Adults could be marked with passive and egg size varies 6 to 10 mm in integrated transponders (PIT tags). Pit diameter. The life cycle is completed tags should be inserted subcutaneously within one season, except in the case of on the dorsum. Juveniles could be over wintering larvae (Nag and marked with different coloured visible Vasudevan 2014). The larvae have well implant elastomer (VIE) but this may developed balancers. The various stages not be necessary for rearing purposes of the life cycle includes egg, early larvae, free swimming larvae, advanced larvae, Housing adults juvenile newt stage and adult. P N H Z P h a s b e e n b r e e d i n g t h e salamander since 2000, and in 2008, Amplexus and Nest Production recommendations were made for a Territorial and breeding behaviour conservation breeding programme with include body elevation, lowering of the PNHZP as the coordinating zoo and head, circling, and even biting (Deuti Himalayan Zoological Park, and Sikkim and Hegde, 2007). Courtship and and Manipur Zoo as participating zoos amplexus mostly occurs at night. (Vasudevan et al., 2014). In order to Amplexus is aquatic and involves the make the conservation breeding male wrapping the forelimbs around the programmes more efficient and f e m a l e s f r o m b e l o w p r i o r t o sustainable, a detailed analysis of the spermatophore deposition and transfer requirements for such a facility must be (see Deuti and Hegde, 2007). created based on geographical data and environmental indicators. Captive Longevity and Age at Sexual breeding will be carried out based on the Maturity strategy described by Ziegler and Lifespan and age at sexual maturity in colleagues (2008) for the closest relative the wild are unknown. species T. shanjing. Chosen individuals from genetic analysis will be housed in Captive Management terrariums measuring 134 cm x 60 cm x 75 cm (length x breadth x height). About Introduction one half of the bottom surface will be As a pre-requisite to a successful and filled with water upto a height of 20 cm long-term conservation breeding with an 8 cm gravel layer for higher program of T. verrucosus, is the design ground. A sheet of glass will be

461 Ex situ Management of Amphibians introduced to divide the terrestrial and in order for the chlorine to dissipate. aquatic portions. The stratum of the Ensure the temperature of the water is at terrestrial section will be formed using equilibrium with the room temperature. moss and tree barks. Small, smooth Water depth should be 80-120 mm, and rocks will be used to create a gradient salamanders should be able to immerse from the water to shore. Some ferns and completely. An emergent rock, marsh grasses will be introduced as vegetation and small flat rocks should be aquatic vegetation. Lighting is provided placed in the water and in the water edge by means of neon tubes and small to allow the salamanders easy access out spotlights. Water and air temperatures of the water. The enclosure should be will be maintained at 18-26°C and 20- lightly misted with aged tap water daily. 25°C respectively. A water filtering and flow arrangement will be installed to Routine Husbandry ensure clean water supply to the All animals should be visually inspected terrarium at all times and pH of the daily. Water dishes must be cleaned out water will be maintained at 8. The and refilled daily as amphibians often breeding season for the Himalayan newt excrete waste products in water. Water is in the start of monsoon around late quality cannot be judged by eye. Faeces April to early May. Slight reduction in and uneaten food must be removed temperature with spraying of water will daily. The enclosure and all furniture be used to provide stimulus for must be thoroughly scrubbed (with a promoting mating (Herrmann, 2001; brush and hot water, no chemical Zeigler et al, 2007). cleaners or disinfectants) twice a week. Temperatures of the enclosure should be checked at different points of the enclosure regularly to ensure that the correct thermoregulatory gradients are maintained. UV readings must be taken every three months to check that there is a sufficient UV gradient and that the UV emitting lights are still emitting UV. The substrate must be changed as soon as it looks dirty or if it becomes saturated or unable to retain humidity. Figure 3. Breeding on-exhibit enclosure at PNZP, Darjeeling (K. Vasudevan) Diet in captivity Basking spots can be provided with Adults have broad unspecialized diet regular incandescent lamps. The and mostly feed on aquatic insects like basking lamp should be positioned at Dytiscid beetles; Gastropod and Bivalve one end of the enclosure and the molluscs (Sphaerium indicum); temperature on the branching below the Decapod crustaceans (land crabs- basking spot should range between 27- Potamon potamiscus sikkimense); 30oC and not exceed 30oC. Ensure that L u m b r i c i d a n d M e g a s c o l e c i d all lighting is inaccessible to the frogs as earthworms; larvae and pupae of they will be burnt easily if they come into Diptera and Coleoptera and numphs of contact with lamps. During the winter Odonata, besides tadpoles of common the basking spot for T. verrucosus amphibians and eggs from foam-nests of should not exceed 22oC. Rhacophorid frogs. Adults sometimes feed on the tadpoles of Bufo verrucosus Check the source of your water by testing and Polypedates teraiensis and also feed it regularly for ammonia, nitrite, nitrate, on snails, slugs, wood-lice and pH and phosphates. Tap water should be coprophilous insects. The larvae are left to stand for 24 hours before it is used bottom feeders and mostly feed on

471 Ex situ Management of Amphibians bacteria, protozoa, diatoms and used to rear larva. Check the source of zooplanktons. Late stage larvae feed on your water by testing it regularly for Chironomid larvae, tubificid worms, soil ammonia, nitrite, nitrate, hardness, pH nematodes and mosquito larvae. and phosphates. Water parameters should be informed by field data but Breeding Seasonality remember that not all breeding sites Salamanders are known to be active may have optimal water quality, from late April to early October, and especially in urbanised environments during this period adults emerge out of where these frogs can often be found. h i b e r n a t i o n a n d m o v e i n t o Tap water should be left to stand for 24 temporary/permanent ponds for hours before it is used in order for the breeding. chlorine to dissipate. Ensure the t e m p e r a t u r e o f t h e w a t e r i s a t Provision of Breeding Sites equilibrium with the room temperature. Adults need leaves of emerging aquatic Check the water in closed systems and vegetation and terrestrial grasses and c a r r y o u t r e g u l a r w a t e r t e s t s sedges over-hanging water on which to (temperature, pH, ammonia, nitrite and lay their eggs. They may also lay eggs nitrate), the results will help you decide among leaves above water in the pond. the frequency of water changes. pH The abundance of the vegetation and should not fluctuate, ammonia should moisture would avoid exposure and measure less than 1 part per million drying up of eggs. (ppm), nitrite less than 0.1ppm and nitrate less than 10 ppm. No more than 20% of the water should be changed as rapid shifts in water quality will be problematic for the biological filtration and larva. Ideally a sump should be used to increase the volume of water of your system and to house all the life support systems (filters etc). If using an internal canister filter ensure that it is not too strong as the larvae could be sucked into the inflow of the filter and may have difficulty feeding. Direct the outflow to the surface of the water for oxygenation.

Figure 4. Egg of T. verrucosus in-situ in Darjeeling (Shruti Sengupta) Husbandry and Development of larva Figure 5: Stage 4 and stage 5 larvae of T. verrucosus from After hatching, larvae can be transferred Darjeeling. to a glass tank with a spoon with utmost caution. When they first hatch they will Substrate is necessary for larva as they not move and it is perfectly normal for like to hide. Dried/autoclaved Indian them to sometimes rest on their side. almond leaves (Terminalia catappa) can The larvae will not start swimming or be added, they also soften the water feeding until this ac has been absorbed. which would be beneficial in hard water The stocking density is important. Tanks areas. Partial (10-20 %) water changes should be stocked with no more than 5 should be carried out regularly, the larvae per litre. Aged tap water can be frequency of water changes must be

481 Ex situ Management of Amphibians governed by frequent water testing. with a maximum UVI of 2. There is no need for basking spot for metamorphs as Larvae should be fed daily once they this will dry them out rapidly and will start swimming freely. They should be likely kill them. A lower temperature fed on a variety of food. Once the front basking spot can be provided with limbs appear and the tail starts to reduce incandescent lamps positioned at one it is important that the metamorphosing end of the enclosure. The temperature salamanders are given the opportunity on the branching below the basking spot to emerge from the water as they can should not exceed 25oC. Small tanks will easily drown. Emerging rocks or aquatic rapidly overheat so ensure that there is plants are suitable. Once they emerge still a cool area in the enclosure. they are prone to drying out and easily Metamorphs should be fed daily until die if the humidity is not high. about six weeks of age with Chironomid larvae, tubificid worms, soil nematodes Rearing Metamorphs and mosquito larvae. All food items Metamorphs can be housed in small should be dusted with appropriate plastic containers. A 350 x 200 x 300 dietary supplements. After 8 weeks the mm enclosure is suitable for up to 12 feeding interval can be gradually individuals, these groups should be increased to once every 3 days. divided amongst larger enclosures as individuals grow. It is important that rearing tanks are relatively small so that high ambient humidity can me maintained and that the juvenile frogs can find food easily. Covering the lid with a fine breathable cloth or fleece will prevent the food escaping and ensure that humidity remains high. The cloth will block dome of the UV emitted from the lights so you must experiment and Figure 6. Metamorph of T. verrucosus in-situ in Darjeeling e n s u r e t h a t t h e U V l i g h t a n d (Shruti Sengupta) temperature gradients are still suitable. Damp soil mixed with sand would make Health a good substrate, and this must be replaced as necessary. A shallow water Problems Encountered in dish must be provided (water depth 10 Captivity mm). It is essential that the water dish is Being a semi aquatic salamander there is easy for the metamorphs to climb out of. potential for limb injuries, skin When transferring metamorphs from abrasions due to objects within the the larva tank make sure that they are enclosure. Periodic scanning of placed in the water dish and not on the individuals for skin lesions should be substrate as they may not be quite ready taken up as a strain of fungus called to leave the water, they will choose when Batrachochytrium salamandrivorans they are ready. Live plants should be (Bs) is known to cause mortality. included in the juvenile enclosures. The metamorphs can be raised at the same Record keeping room temperature as the adults (20 - 25˚C), and must be misted with room Things to include. temperature tap water twice daily. It is Individual identification, weight, vital that metamorphs and juveniles are feeding records, temperature records, provided with UV at this critical growing UV records, water quality test results, stage (Arcadia D3+ reptile lamp T5 12% h e a l t h r e c o r d s , b e h a v i o u r a l o r UVB), this must be given in a gradient reproductive observations.

491 Ex situ Management of Amphibians

Bibliography a population of Tylototritonverrucosus (Amphibia: Salamandridae) from the Anders C., Schleich H., Shah K. 1998. Himalayan Region. Copeia, 2010(4): Contributions to the Biology of 600-608 Tylototriton verrucosus Anderson 1871 from East Nepal (Amphibia: Caudata, Vasudevan K., and Nag, S. 2014. Salamandridae). In: H Schleich and W Observations on overwintering larvae of Kastle (eds.) Contributions to the Tylototriton verrucosus (Caudata: Herpetology of South Asia (Nepal, S a l a m a n d r i d a e ) i n D a r j e e l i n g , India) No. 4. Fuhlrott Museum, Himalaya, India. Salamandra 50(4): Wuppertal, Germany, 1-26 245-248.

Deuti, K. and Hegde, V.D. 2007. Vasudevan K., Kumar S. and Sengupta S. Handbook on Himalayan Salamander – 2014. Status survey and conservation of Nature Books India: New Delhi, India the Himalayan Crocodile Salamander Tylototriton verrucosus in the Eastern Kuzmin, S. L., Dasgupta R., and Smirina, Himalayas. Final Report. Wildlife M. 1994. Ecology of the Himalayan Newt Institute of India, Dehradun pp.31. (Tylototriton verrucosus) in Darjeeling Himalayas, India. – Russian. Journal of Ziegler, T., Hartmann, T., Straeten, K. Herpetology, 1(1): 69 – 76. Van Der, Karbe, D., and Bo, W. 2008. Captive breeding and larval morphology Schleich H. H., Kästle W. 2002. of Tylototriton shanjing Nussbaum , Amphibians and reptiles of Nepal. Brodie and Yang , 1995 , with an updated Biology, Systematics, Field Guide. key of the genus Tylototriton (Amphibia: Germany: A.R.G Gantner Verlag K.G. Salamandridae), Der Zoologische Garten 77: 246-260. Seglie D., Roy D., Giacoma C. 2010. Sexual dimorphism and age structure in

501 Ex situ Management of Amphibians

9. Marking of Amphibians

To study large number of amphibians in captivity, needs a reliable technique to identify individuals within populations. Traditionally – toe clipping, freeze and hotwire branding, tattooing & various methods of tagging have been used. These techniques were painful to the animal and had disadvantages too. Thus no longer are recommended.

With the recent developments in technology & looking into the animal welfare aspects, as well as their current acceptability, Passive Integrated Transponder Tags (PIT), permanent method of marking are being used, however it has several advantages and disadvantages too. These methods have been described in brief:

Passive Integrated Transponder Tags (PIT)

Figure 2 Reading the Passive Intergrated Transponder To mark the amphibians with PIT, it is Number. (B.K. Gupta) injected into the left caudal body cavity – • Requires handling and tissue Subcutaneous – into the dorsal penetration lymphatic sacs of the frogs. • Limited to use in larger frogs – Animals in lower allowable size class (40-50 mm snout vent length [SVL]) occasionally show Passive Integrated Transponder Tags (PIT)

• PIT tags should only be used for frogs >50mm SVL.

Visible Implant Fluorescent (VIE) Figure 1 Passive Integrated Transponder insertion in a frog Elastomer Tags (B.K. Gupta) VIE tags are silicon-based components - Passive Integrated Transponder mixed just prior to use A range of Tags (PIT) Advantages: colours and injection sites combine to produce a large number of individual • Unlimited number of codes codes. This technique has been used on tadpoles, as well as adult frogs with some • Painful success.

• High cost

511 Ex situ Management of Amphibians

Pattern Mapping

Advantages

• Low cost - unless software needs to be purchased

Figure 5. Pattern mapping in Amphibians (DWCT) Figure 3 Tadpole marked with visble implant elastomer (DWCT) • Technology still being developed, i.e. pattern recognition software

• Limited by numbers – cumbersome with a large population

• Time consuming

• Potential temporal shifts in pattern. The application of pattern mapping is limited by the number of species that display unique individual markings.

Figure 4. Marking with Alcian blue dye solution using a "panjet" innoculator (DWCT) • Allows individual identification of very small frogs, juveniles

• Non invasive

• No risk of infection or spread of disease

• No pain

Disadvantages

• Useful only in species with unique individual markings

• Need to handle frogs (as with most techniques)

521 Ex situ Management of Amphibians

10. Culturing food for Amphibians

Any captive bird, mammal, reptile or as spent females will dig up and eat your amphibian that would naturally include eggs! insects in their diet will benefit from a variety of good quality invertebrate foods.

Attention to this aspect of diet and nutrition can make the difference between captive animals simply surviving in adequate health or thriving and breeding successfully. However providing the food for a captive breeding effort may demand more physical time than the breeding animals themselves; Figure 1 Indian House Crickets (Wikipedia) insect culturing is intensive, dynamic and often time consuming. The laying medium should be selected for humidity holding and may be As per the requirement of animals and compost, topsoil, sand, peat etc suitability of its culutre, we may identify suitable species of invertebrates, with the following objectives: - R e p r o d u c i b l e w i t h g o o d cost/benefit ration - Gregarious - Easily identifiable - Non toxic - Various sizes for the species they are intended for - Found naturally in large enough numbers to make gathering founders, a feasible job

Culturing Indian House Crickets:

Indian House Crickets can be cultured Figure 2 Culture box with used egg trays (DWCT) easily in zoos. For culturing them new colony may be established by starting Crickets can be fed with fresh fruits with housing of few adult crickets. (oranges, papaya) and vegetables (cabbage, greens) and a home made Up to 3000 adult field crickets can be cricket cake-diet made of oat powder, comfortably housed in a smooth sided skimmed milk powder and sugar. container measuring approximately; 100cm (l) x 80cm (w) x 80cm (d) To ensure that all individuals in the colony have had sufficient opportunity Suitable sized nest boxes (20 x 20 x to feed in each 24 hour period, enough 10cm) must be supplied to enable eggs to food needs to be offered to allow an be harvested and removed to a separate excess on removal. hatching tub. The nest boxes must be covered with fine metal mesh (0.5cm sq) The amount of food offered should be 531 Ex situ Management of Amphibians adjusted on a daily basis. Adding large Cultures of fruit flies are difficult to get amounts will lead to mould and wastage, consistent and must be carefully as well as wasted time preparing the managed and adjusted to ensure food, adding the food and removing the success. The cultures can smell pretty food! bad as fermentation of the substrate is a necessary part of the process! Dried foods should comprise the bulk of the diet and do not need to be removed once added. Care must be taken to add just enough to prevent moulding.

Protein content is key, crickets need between 25-30% in order to be robust and productive breeders.

Fresh foods can be scraps, peelings, herbs, greens, forage or whole fruits and veg. Anything non-toxic that will provide moisture without going soggy or Figure 4 Fruit flies culture containers (DWCT) mouldy within 24 hours will do. The fruit mix is the most nutritious for Basic choices are orange, potato, carrot, the flies, providing the best variety of sweet potato or apple. vitamins .

Experimenting for variety is a good idea This substrate produces a fast life cycle but avoid soft fruits such as papaya, in the flies, the culture is ready for tomato or plum these will leave your feeding out 2weeks after it is made, but colony with a big hygiene problem. only normally lasts 7-10 days after the initial peak activity. The most difficult aspect of culturing crickets is maintaining a high level of hygiene and a standard procedure to run your colony “like clockwork”

Temperature, humidity, density, feeding and hygiene must all be maintained to a high standard.

Culturing Drosophila Fruit flies are prolific breeders and cultures do not take up much room. Figure 5 Mixture used for culturing the fruit flies (DWCT) Fruit flies are ideal food for a huge The recipe from Drosophila culture; number of species of amphibians. - 2 ripe bananas

- 1 ripe papaya

- 2 pears

- 2 apples

Figure 3 Drosophila (DWCT) - 2 carrots 541 Ex situ Management of Amphibians

- multivitamin supplement A piece of cardboard egg tray, wood shaving etc. is used as a platform for the - brewer's yeast breeding flies themselves.

- oats Each jar should have just 15 Drosophila added to begin a new culture The mix should not be so dry that fly larvae cannot move through it, or so wet Finally a double layered cloth lid is that it falls out when you try to harvest needed to prevent wild flies breeding flies… this is the hard part! with your flightless cultures.

The mix should be evenly distributed between disposable pint cups, glasses, pots etc.

Glass viewing amphibian exhibits with natural vegetaon cover. (Brij Kishor Gupta)

551 Ex situ Management of Amphibians

11. Enrichment of Amphibian Exhibits

Today modern zoological facilities strive improve captive breeding success. to provide for the complex needs of animals and aim, as far as possible, to reproduce the natural environment and habitat which should meet the natural needs of the species in their care. However, those who care for wild animals in zoos understand that for many animals, life in a zoo, no matter h o w w e l l m a n a g e d , i n v o l v e s compromise. This does not mean that the welfare and care of animals in zoos Figure 1 Terraium type, glass fronted exhibit for ampbhibians are necessarily compromised, and does with live vegetation. (Brij Kishor Gupta) not minimize the ongoing efforts to meet the needs and domains of animal welfare Unfortunately, as little is currently for all species. known about the micro-climates utilised by most amphibian species in the wild The high quality of care provided in a keepers may have to collect these data captive environment significantly for themselves or correspond with field reduces the time an animal must spend biologists in order to obtain these very in these pursuits, but does not address important data. the behavioural needs associated with these activities. It has become necessary Enclosure to provide alternative methods of Enclosure dimensions will depend on stimulating natural behaviour of the the type of species being kept and species housed therein to meet both the typically larger and more active mental and physical needs of the captive amphibians require larger sized animal. Enrichment has become enclosures. Arboreal species should be essential in zoos to promote species- provided with tall enclosures (E.g. typical behaviour by providing animals Polypedates spp. Rhacophorus spp. with a naturalistic environment as of Pedostibes tuberculosus) and terrestrial similar to wild situations. That species (e.g. Duttaphrynus spp. environment may include activities that Fejervarya spp. Clinotarsus curtipes) a r e b o t h c h a l l e n g i n g a n d t i m e should be provided with a greater consuming and may serve several amount floor space; enclosure height is functions, for achieving so, we should less important for these. Note that some know following: species can be incredibly active and jumpy and when they are maintained in Know your species small enclosures they may be prone to Amphibians have complex and varied rostral abrasions. husbandry requirements and are not always easy to maintain and breed. In captivity keepers should aim to replicate w i l d m i c r o - e n v i r o n m e n t s a n d conditions as closely as possible and where feasible captive management should be informed by field data as well as knowledge of the biology of the Figure 2 Open, large, naturalistic breeding cum exhibit area species. Replicating wild conditions may for Himalayan salamander at PNHZ Park, Darjeeling (Brij Kishor Gupta) 561 Ex situ Management of Amphibians here are several important factors to burry into. The humidity of the substrate consider when selecting an enclosure for and its potential of water absorption and amphibians. Enclosures must be escape retention also needs to be considered. As proof. Due to their permeable skin amphibians have permeable skin (and amphibians may quickly desiccate and drink through their skin) there should be die if they escape. The construction a moist area of substrate at all times. material should be easy to clean and preferably be smooth. Wood is not appropriate as it may warp with high humidity. It is not possible to disinfect wood during routine cleaning so this material should be avoided. Many chemicals used to seal wood are toxic to amphibians. Plastic enclosures are a suitable alternative. Any plastic should be food grade as many plastics can leach substances that may be detrimental to amphibian health over time. Plastics may also become brittle when exposed to Figure 3 Acrylic fronted, terraium type exhibit for amphibians U V B r a d i a t i o n ( a n i m p o r t a n t with natural logs and sand as substrate at Coimbatore Zoological Park (Brij Kishor Gupta) component of amphibian husbandry). Plastic can be easily cleaned and A gradient of humidity in the enclosure disinfected. Glass is another alternative; should be provided which will allow the it is readily available, easy to clean and amphibian to self-regulate its water disinfect although glass may break balance. For the majority of species easily. Construction of enclosures using water logged substrates are not optimal. brick and cement should be avoided as Amphibians do need to be given the these materials may leach minerals into opportunity to sit in dryer areas within water bodies and may be detrimental to the enclosure and failing to provide w a t e r q u a l i t y . A m p h i b i a n s a r e some dry areas could result in serious ectothermic and require a gradient of health issues. Substrates should be UVB radiation, temperature, visible changed as and when needed. Some light and humidity, therefore amphibian amphibians may ingest substrate; this is enclosures will need to be lit and not always a problem unless the appropriate sized meshed apertures amphibian ingests something it is need to provide air exchange in the roof u n a b l e t o p a s s t h r o u g h i t s or lid of the enclosure. Keeper access is gastrointestinal tract (e.g. pebbles / also an important factor to consider for strings of moss), the feeding behaviour ease of maintenance while minimising of the amphibian should therefore be chances for escapes while servicing the considered when selecting a substrate. enclosure. Ideally, drainage holes with Some substrates can be very powdery taps will need to be provided to facilitate and may stick to the skin of an easy cleaning. amphibian and interfere with the function of the skin (gaseous exchange Substrate etc.). Substrates for surface active There are a huge variety of substrates amphibians should therefore be available for amphibians and the type of compacted to minimise this. substrate used will depend on the species being maintained. It is Refugia / furnishings important to consider the depth of the All amphibians should be provided with substrate. Fossorial amphibians (e.g. refugia. The provision of multiple caecilians, Sphaerotheca spp.) may refugia within an enclosure will make require deep substrates for them to the enclosure more heterogeneous. This 571 Ex situ Management of Amphibians is of fundamental importance as due to their differing life histories and amphibians will select sites to regulate micro-habitats. UVB is important in the their temperature and water balance. synthesis of vitamin D3. Vitamin D3 Cover objects and visual barriers are also deficiency is a nutritional problem important, amphibians are eaten by a encountered in captive amphibians. wide variety of other organisms and Vitamin D3 plays a critical role in keeping them in very open enclosures regulating calcium metabolism, as well with limited cover can result in severe a s i m p o r t a n t r o l e s i n m u s c l e rostral abrasions, stress and disease. For contraction, organ development and the hides to have the desired effect of functioning of the immune and nervous providing adequate humidity and to systems. In the majority of vertebrates, reduce stress, they will need to be vitamin D3 is synthesised via exposure individually selected to fit the target to the ultraviolet B radiation (UVB) species' size and shape so that the present in sunlight. The synthesis of animals fit in or under snugly. Arboreal vitamin D3 in the skin requires heat, species will require branches as perch / therefore heat must be provided in rest sites and some species will be tandem with UVB radiation, matching b e n e fi t f o r m l i v e p l a n t s ( e . g . temperatures found in the wild. Some Rhacophorus spp.) which often perch on amphibian species might also receive leaves. Aquatic species (including vitamin D3 through some diets although tadpoles) will also been shown to benefit the extend of this is largely unknown; from refugia. Ensure that any live plants some food supplements can provide are obtained from sources free from vitamin D3 artificially although great agrochemicals. Also make sure that care must be taken to achieve the correct plants do not have thorns, irritant or dosages and that the supplement is toxic sap, hairy leaves/stems or other stored correctly and stock is frequently features that may harm amphibians. replaced. Dietary supplements are Additionally, food insects may feed on degraded by light, humidity and heat. plants in terraria before being consumed, so ensure that plant toxins cannot be ingested by amphibians by this route.

Light and temperature Wild amphibians have evolved in accordance with their environments. They will reconcile requirement and provision by self-regulating their exposure to heat, light and the Figure 4 Amphibian house at Jersey Zoo, Jersey, Channel associated UVB radiation. Optimal Islands, United Kingdom. (Brij Kishor Gupta) UVB, light and temperature levels for a species will be those in their natural In order to mimic the situation in the habitat, specifically at the microhabitat wild, heat (infrared radiation), light and level for each life-history stage. Where UVB should be provided together (i.e. possible the UVB, light and temperature the highest levels of UVB should be microclimate throughout the year associated with the warmest areas of the should be established through field enclosure, and the lowest levels with the studies and used to inform the captive coolest areas). Most importantly, all management of the target amphibian three must be provided as gradients in species. The UVB and Vitamin D3 the enclosures for appropriate lengths of requirements of amphibians are largely time each day in order to facilitate self- unknown. It is likely that different regulation, with retreats that provide species will have different requirements deep shade and no direct UV radiation.

581 Ex situ Management of Amphibians

If there are life-history stages that installed to cool water in enclosures. would, in the wild, only be exposed to low-level reflected or diffused UV-B, or Maximum and minimum temperatures none at all (e.g. eggs and tadpoles in tree in amphibian enclosures in both the cool holes or burrows), then care should be end and warm end of the enclosures taken to replicate this situation in should be recorded on a daily basis. The captivity. The provision of appropriate UV index should also be recorded to UV-B radiation to captive amphibians is ensure that UVB provision is adequate. important for their health and proper The UVI index can be measured using a development but this field is still in its Solarmeter 6.5. infancy. We strongly advise caution in its provision as over-exposure can also cause harm. We believe that tolerances to UV-B radiation will be species- specific and related to the historic exposure of animals; individuals that have not been provided with UVB in the past should be slowly exposed to increasing levels until reaching optimal exposure to allow the skin to adapt to Figure 5 Himalayan salamanders provided with wet mud radiation. as substrate at PNHZ Park, Darjeeling. (Brij Kishor Gupta)

Amphibians should not be able to come into direct contact with any lamps or heaters as they have extremely sensitive skins and may be easily burnt. All lamps will emit heat but additional heating and cooling may also be necessary. Rooms can be heated and cooled with air c o n d i t i o n i n g u n i t s . A m p h i b i a n enclosures should not be placed under such units, or normal fans, as the air m o v e m e n t c a n c a u s e s e v e r e Figure 6 Young himalayan salamanders also provided with boulders as substarte and algae as feed at PNHZ Park, dehydration. Chilling systems can be Darjeeling. (Brij Kishor Gupta)

Water quality Recommended levels Control methods parameter Water hardness (dissolved For soft water amphibians: <75mg litre-1 Soft water: Ca and Mg salts) (ppm) of CaCO3. Harden using Ca and Mg salts (only For hard water amphibians: >100mg litre-1 recommended reconstituting RO (reverse of CaCO3. osmosis), DI (de-ionized) or distilled water). Hard water: Soften using RO, DI or distilled water.

Dissolved oxygen as O2 >80% saturation Aeration Gas supersaturation Gases maintained at equilibrium with the Aerate water until equilibrium with atmosphere atmosphere. is achieved.

Ammonia/ Ammonium- <0.2mg litre-1, N as unionized ammonia Biological filtration, chemical filtration (with + NH3/NH4 appropriate medium), water changes. - -1 Nitrites NO2 <1.0mg litre , ideally 0 Biological filtration, chemical filtration (with appropriate medium), water changes. - -1 Nitrates NO3 <50.0mg litre Removal: photosynthetic action of green plants or water changes.

pH Generally near neutral, although it is Change water source or add appropriate buffer species-dependant. Should avoid pH <6 solution. and >8. Chlorine Cl2 0 Aerate for 24 hours, or add chemical dechlorinator such as sodium thiosulphate. 591 Ex situ Management of Amphibians

-1 Chloramines (CINH 2, <0.01mg litre as CI Use chemical treatment specific for CIN2H, CIN3) chloramines such as Prime® (Seachem Laboratories,Inc., Madison, GA 30650, USA). Filters for this purpose are available. Copper (Cu) <0.05mg litre Carbon filtering and carbonate precipitation (do not use copper piping).

3 - Posphates (PO 4 ) Toxicity species-specific; EPA Lower levels of phosphates using phosphate recommends limit of 10mg litre-1; 1mg sponges and filters. litre-1 is effective for preventing pipe corrosion.

Water quality parameters: guidelines for be treated with caution; avoid using keeping amphibians. Adapted from water which has previously been in Odum & Zippel (2008). contact with other amphibians as this may risk disease transmission, or may Water have been contaminated by dissolved substances. Reverse osmosis (RO) Water quality is an extremely important systems will produce water that is too component of amphibian husbandry. pure for amphibians and using pure RO Monitoring water quality is vital to may cause osmoregulatory imbalances. successfully rearing healthy captive RO water can be made suitable either by a m p h i b i a n s ; fl u c t u a t i n g w a t e r adding salts or a known quantity of tap parameters create stress for the water. Avoid water that has been stored individuals, therefore it is better to in metal tanks or exposed to any other maintain constant conditions, even if substances that could leach into the these are slightly sub-optimal – of water. Low levels of dissolved metals can course consistent optimal water quality be toxic to amphibians. should always be the goal. Water parameters should be tested frequently in newly established systems, and tests can become less frequent as the system develops. Testing should be carried out on a regular basis in order to monitor the effectiveness of biological filtration.

The most common system used for keeping aquatic and semi-aquatic animals is a semi-closed system, in which a combination of filtration and Figure 7 Water quality is extremely important for all amphibians housed at the zoo. (Brij Kishor Gupta) water changes are employed in order to maintain water quality. Water changes It is important to know certain should be small and regular (no more parameters of the water source; sources than 20% of the original volume of the can be tested for dissolved substances, water body in question). Rapid changes pH and hardness, and treated if in water parameters can be highly necessary to provide the appropriate stressful on the physiology of aquatic conditions. Unfortunately the optimal organisms. The necessary frequency of water parameters are unknown for most water changes will depend on the amphibian species. Most institutions stocking density, the temperature of the use municipal water sources which are water and the amount and type of food generally treated with chlorine, or given at each feed, as these have an effect chloramines; levels of these chemicals on the level of activity within the tank, should be tested for and treated which in turn affects the amount and accordingly. Other water sources should character of waste material produced. 601 Ex situ Management of Amphibians

Suspended particles can be removed Mechanical filters remove physical from the water column through particulates from the system, and these mechanical filtration, dissolved harmful then build up in the filter if they are not substances (such as chlorine) through cleaned out straight away. Build up chemical filtration, and nitrogenous should be avoided as it will reduce the waste material produced by aquatic capacity of the filter for dealing with organisms and the breakdown of organic further suspended particles, and any material can be processed by bacteria organic matter within them may begin to living on filter media, through the b r e a k d o w n a n d l e a c h h a r m f u l process of biological filtration. This can substances back into the system. be achieved through using separate filters, or single filters with combined Chemical filters are designed to remove functions. The outflow of the filter dissolved substances such as organic should be adjusted to meet the compounds from the water as it passes requirements of the amphibian. Some through; activated carbon has been tadpoles require extremely fast flowing identified as an ideal material for well oxygenated water whereas this may amphibian systems and is commonly be detrimental to other tadpoles (e.g. used as a chemical filtration medium. It those living in ponds). is vital that such filter systems are used in combination with a mechanical filter, as build-up of particulate matter will severely compromise their effectiveness. Chemical filter media will need to be routinely replaced. Be aware that a c t i v a t e d c a r b o n w i l l r e m o v e d medication and tannins from the water column, so may be inappropriate for s o m e s y s t e m s a n d u n d e r s o m e circumstances.

Biological filtration systems consist of a community of living organisms which Figure 8 Water parameters should be monitored by testing frequently in new established exhibits. (Brij Kishor Gupta) act to break down toxic metabolic waste products in the water to less toxic The size and type of filter used will substances; toxic ammonia is broken depend on the size of the enclosure, down to less toxic nitrite which then in volume of water and density of turn is converted to less toxic nitrate individuals being catered for. If larvae (which in itself is still less toxic to aquatic are being kept at relatively high organisms). These filters are therefore densities, large external filters may be the most important component in necessary to remove waste material; for maintaining the health of the aquatic those kept at lower densities (under 20 system and the amphibians themselves. individuals per litre), box filters may be Owing to the living component of this adequate. Filters require regular filtration system a constant supply and maintenance in order to ensure they flow of water is required in order to continue to function effectively. The ensure nitrogenous waste is absorbed frequency with which filters need and that oxygen is available to the servicing depends on the stocking bacterial community. Biological filters density, the amount of food given per take time to mature and can be seeded individual per feed, as well as the with filter media from established temperature of the water, as this will systems, this may risk introducing affect how much waste material is disease causing agents into new set ups. produced. Alternatively filters can be matured

611 Ex situ Management of Amphibians using an ammonium chloride dosing Even when the diet is known, it is often regime. Note that filters are less efficient impossible to replicate in captivity as at processing nitrogenous waste in soft captive diets are limited by the water and at low temperatures. commercial availability of food species and the ability to establish breeding Consideration must be given to the colonies of appropriate species, as well access points in and out of the water as difficulties in providing prey species body. Many amphibians are surprisingly themselves with suitable diets. poor swimmers and can easily drown. Nevertheless, nutrition is a key factor in keeping amphibians healthy and a basis Plants for successful breeding. Great efforts Plants are beneficial in amphibian should be made to be able to provide any enclosures. Not only do they provide captive amphibians with as much variety resting sites for amphibians but they and good nutritional content as possible. also create microhabitats (e.g. humid pockets of air caused by transpiration and also shade from heat and light) as well as oviposition sites for some a m p h i b i a n s p e c i e s . P l a n t s a r e particularly important in aquatic exhibits as they will use the nitrogenous compounds for growth, as well as absorbing many other pollutants, and can improve water quality. Figure 10 Feed for mansy of the species of amphibians. (Brij Kishor Gupta) Diet Most post metamorphic amphibians are Nutritional problems are a major barrier insectivores and will only feed on live, to successful amphibian husbandry. The moving prey items. Invertebrates can be nutritional requirements of most cultured or collected from the field. Food a m p h i b i a n s a r e u n k n o w n , a n d offered to amphibians should be no requirements change with life stage for bigger than the width of the head. All species with tadpoles. food should be well fed prior to being offered to the amphibian, in addition to being dusted with an appropriate dietary supplement. To encourage natural feeding behaviour amphibians should be fed at times to coincide with their peak activity periods to ensure that they encounter the food and that dietary supplements are still coating the prey item when the food is ingested.

Seasonality and breeding Many amphibians breed in response to environmental cues, these include, but are not limited to temperature, humidity, precipitation, barometric pressure and changes in water quality. It is important that seasonal changes that reflect the situation in nature are replicated in captivity even if breeding is not the desired outcome as failing to Figure 9 Culturing crickets (Courtesy: DWCT) provide animals with the seasonality 621 Ex situ Management of Amphibians may cause health issues e.g. females Routine maintenance becoming egg bound, obesity etc. Faecal material and dead feeder insects should be removed from enclosures on a daily basis. Any uneaten food should be removed the day after the animals have been fed as the nutritional value of the food will have deteriorated during the time it is has been in the amphibians' enclosure. Feeding regimes will depend on the species being maintained, the season and life stage of the amphibian. Water dishes should be changed daily and the same attention should be paid to the water source for water dishes as for

Figure 11 The amphibians should be provided species aquaria. Amphibians often excrete large specific substrate during the period season to meet their volumes of urine whilst sitting in water biological and behavioural requirement.(Brij Kishor Gupta) and even if the water appears clean it may not be. Captive amphibians Rain chambers are one of the most (particularly non-native species) may common ways to breed amphibians in carry pathogens that are not present in captivity. These systems recreate rain the vicinity of the amphibian holding showers and usually involve a small facility. Waste water from amphibian pump sitting in a few centimetres of enclosures should be disinfected water, the pump supplies water to a rain following the manufacturer's guidelines. bar. The frequency and duration of the Temperatures should be checked and rains can be controlled by using a timer. recorded daily and the function of all aquatic life support systems lights and heaters checked on a daily basis. Special attention should be paid to humidity gradients within the enclosures, if an area has become too dry is can be manually sprayed with water from an appropriate source. Ideally amphibians should be given a visual inspection on a daily basis. Unearthing fossorial species such as caecilians and some frogs may be detrimental to their health and welfare so disturbance should be minimised for these species.

Figure 12 Amphibians whose egg laying patter varies, should be provided appropriate artifacts as of to meeting the species requirement (Brij Kishor Gupta)

631 Ex situ Management of Amphibians

12. Amphibian Diseases

Introduction: fungus infection in Asia. Nearly 520 frog species have been affected by Bd fungus Amphibians are undergoing mass and population decline has been mortality due to various pathogens that observed in more than 200 species include viruses, bacteria, protist worldwide (Olson et al., 2013; Fisher et metazoans, water molds and fungus. al, 2009). Rana virus disease outbreak in wild as well as captive population has led to Bd has two stages, reproductive mass mortalities in different parts of the zoosporangium and motile zoospores. w o r l d . B a c t e r i u m , A e r o m o n a s Zoospores which are motile colonize and hydrophila is responsible for red legged encyst on the surface of frog skin disease in Rana mucosa. Fungal epidermal cells and penetrate into inner p a t h o g e n B a t r a c h o c h y t r i u m epidermal cell line and develop into dendrobatidis (Bd) has caused more thallus. This thallus matures and damage to amphibians than any other produces sporangia which release new known pathogen, and it is responsible zoospores. This process causes for global declines in their population. hyperkeratosis in frogs. The skin This is most common disease and it is becomes thick-walled, which interferes reported from the Western Ghats in with gaseous exchange, osmoregulation India, therefore, we focus here on Bd and electrolyte transport which leads to fungus. cardiac arrest (Voyles et al, 2009).

Batrachochytrium dendrodatidis Very few treatment options are (Bd) is an aquatic pathogenic fungus available for Bd fungus. Previous studies that is capable of infecting the frog skin have shown that several symbiotic and disrupting osmoregulation bacteria have ability to inhibit the resulting in chytridiomycosis. A recently growth of Bd fungus. Bacteria can d i s c o v e r e d s p e c i e s o f f u n g u s , produce anti-fungal compound like Batrachochytrium salamandrivorans Violacein against Bd fungus. Heat ( B s ) , i s a l s o k n o w n t o c a u s e treatment is another treatment option chytridiomycosis in salamander and which can cure Bs infection. It is newts (Martel et al, 2013). Bd and Bs therefore necessary for effective fungus are closely related and have surveillance to be able to detect the similar etiology. Bd probably originated disease at its early onset in order to from Africa and spread throughout the successfully manage them in captivity. continent and elsewhere (Weldon et al, 2004). Trading of Xenopus frogs for Swabbing for Bd/Bs fungus medical and food purposes resulted in detection: It is is non invasive and transport of the fungus to different sensitive. A fresh pair of plastic or latex countries. Bd fungus has reportedly hand gloves (NOT powdered gloves, as it caused mass mortality in America, will remove moisture from the skin of Australia, Africa and Europe; but in Asia frogs) should be used for handing each the prevalence is low. It is suspected that frog. For each frog, take a new sterile chytrid infection is emerging in Asia cotton swab. A total of 70 swab stroke (Swei et al, 2011). So far, only Japan, from the frog's body should be taken as China, Indonesia, Hong Kong, South shown in figure 1 (Kriger et al, 2007). For Korea and India have reported Chytrid each salamander, take 5 strokes

641 Ex situ Management of Amphibians fromeach limbs, 10 strokes from its belly keep the frog in captivity and report the and 5 strokes along the bottom of tail. symptoms immediately to experts to get Used gloves should be disposed using t h e i r o p i n i o n standard protocols, incineration is a [http://www.amphibians.org/asg/me preferred method. After sampling, mbers/]. instruments and boots should be washed with alcohol or Sodium Supporting data required for hypochlorite. During sampling, swabs swabs: The following information can be stored in ice box and later it needs to be gathered and recorded for should be transferred to -20°C. each swab sample for processing the lab.

Clinical symptoms: Common sings of a) Serial No: Start with Your initials Bd includes discoloration and reddening and 0001 (For example, if your name is of frog (fig. 2) skin colour, unusual Robert Frost then your first sample postures like keeping belly away from should be labelled RF0001) ground, hardening of skin, excessively peeling of skinand unusual behaviour b) Sample ID: This is a combination of like nocturnal frogs becomes active columns A, B and C. First two alphabets during day time. Bs is known to infect of the C and B and the number on A will only salamanders, where they show be the Sample ID deep ulceration on skin (fig. 3). In some c) Name of collector: Please enter other cases, at sites with low prevalence your full name frogs/ salamander might be found dead. In order to diagnose the disease and d) Location: As accurately as possible. testing swabs in a lab is essential. GPS location is preferred.

Diagnosis: It can be done by qPCR e) Altitude: Elevation in meters if which is more sensitive method than available, if not try to give an estimate. It n o r m a l P C R a n d h i s t o l o g i c a l could be a range. procedures. qPCR is usually carried out using Bd specific Taqman MGB probe f) Max and Min Temp: The maximum following standard laboratory protocols. and minimum temperatures on the day Molecular diagnosis for Bd/Bs can be of collection in degrees Celsius, if done at Centre for Cellular and available. If not remember to give a Molecular Biology, Hyderabad [see range of temperatures. website:http://www.ccmb.res.in/lacon es]. As set of 10 swab samples can be sent g) Date: Day, Month and year as per the for analysis. A report on the level of collection infection in the samples will be conveyed to the concerned persons. h) Habitat: please use what works but whatever you do make sure that you Mass mortality: Andy dead frog provide us with a legends and that you should be photographed in different remain consistent across all your angles and put in separate containers samples. and stored in deep freezer immediately. Necropsy can be performed using standard procedure and tissue samples (For example, secondary forest or can be sent to laboratories for forensic eucalyptus plantation, or riparian zone) analyses. If you encounter a sick frog Since there are many possibilities, make having symptoms mentioned above around 5-6 types and then roughly distribute samples into them. 651 Ex situ Management of Amphibians i) Genus: Give genus of species. Take a k) Snout-Vent length: Give the value photograph of the frog to help in of length from the snout to the cloaca of identification. the frog in mm or cm j) Species: Give species name if sure, if l) Weight: If possible weigh the frog and not say ~ and whatever you think the present data in grams. species might be. In such cases try to take good pictures of the individual and m) Skin lesions/deformities: If send to experts for identifying. Please present, type "yes" and give details, if not donot try to guess. This is not a just type in "no". See Figure 4. competition.

Fig.1. Pictorial dipiction of frog swabbing and number of strokes as mentioned

661 Ex situ Management of Amphibians

Fig.2: Bd infected frog showing reddening of legs

Fig.3: Bs infected salamander (Salamandra salamandra) (A) infected salamander shows patches on skin (B) Uninfected salamander - Adapted from Blooi et al, 2015

Fig.4: Emaciated or sick frogs A) Fejervarya sp with a red transparent streak on the skin; B) Limnonectes sp with a transparent blackish blotch on the skin; C) Shrunken waist or dehydrated frog (dorsal view); D) Fejervarya sp with grayish blue transparent streak on the skin. 671 Ex situ Management of Amphibians

References dendrobatidis, the amphibian chytrid fungus. PloS One. 8 (2), 1. Blooi M, Martel A, Haesebrouck F, e56802 Vercammen F, Bonte D, Pasmans F. (2015) Treatment of urodelans 6. Swei A, Rowley JJL, Rödder D, based on temperature dependent Diesmos MLL, Diesmos AC, et al. i n f e c t i o n d y n a m i c s o f (2011) Is Chytridiomycosis an B a t r a c h o c h y t r i u m s a l a m Emerging Infectious Disease in andrivorans. Scientific Reports 5, Asia? PLoS ONE 6(8): e23179. A r t i c l e n u m b e r : 8 0 3 7 . doi:10.1371/journal.pone.0023179 doi:10.1038/srep08037 7. Voyles, J., S. Young, L. Berger, C. 2. Federici E, Rossi R, Fidati L, C a m p b e l l , W . F . V o y l e s , A . P a r a c u c c h i R , S c a r g e t t a S , Dinudom, D. Cook, R. Webb, R. A. M o n t a l b a n i E , F r a n z e t t i Alford, L. F. Skerratt, Speare R. A, La Porta G, Fagotti A, Simoncelli ( 2 0 0 9 ) . P a t h o g e n e s i s o f F, Cenci G, and Di Rosa I. (2015) chytridiomycosis, a cause of Characterization of the Skin catastrophic amphibian declines. Microbiota in Italian Stream Frogs Science 326:582–5. ( R a n a i t a l i c a ) I n f e c t e d a n d Uninfected by a Cutaneous Parasitic 8. Weldon C, Preez L, Hyatt A, Muller D i s e a s e . M i c r o b e s E n v i r o n . R, Speare R. (2004) Origin of the Vol. 30, No. 3, 262-269 A m p h i b i a n C h y t r i d F u n g u s . Emerging infectious diseases. Vol. 3. Fisher, M. C., Garner, T. W. J. & 10, No. 12 Walker, S. F. (2009) Global emergence of Batrachochytrium dendrobatidis and amphibian C h y t r i d i o m y c o s i s i n s p a c e , time and host. Annual Review of Micriobiology. 63, 291-310.

4. Martel A, Sluijs A, Blooi M, Bert W, Ducatelle R, Fisher M, Woeltjes A, Bosman W, Chiers K, Bossuyte F, P a s m a n . ( 2 0 1 3 ) Batrachochytrium salam and rivora s s p . n o v . c a u s e s l e t h a l chytridiomycosis in amphibians. P.| Natl. Acad. Sci. USA 110, 15325–15329

5. Olson, D. H., Aanensen, D. M., Ronnenberg, K. L., Powell, C. I., Walker, S. F., Bielby, J.,Garner, T. W., Weaver, G. & Fisher, M. C. ( 2 0 1 3 ) M a p p i n g t h e g l o b a l emergence of Batrachochytrium

681 Ex situ Management of Amphibians 13. Prioritized Species

List of target species and practice species with the name of zoological parks identified during the workshop for initiating the amphibian housing, exhibit and if required planned conservation breeding in India.

Target Species* 1. Nasikabatrachus sahyadrensis (WG) 2. Melanobatrachus indicus (WG) 3. Tylototriton verrucosus (NE) 4. Scutiger occidentalis (H) 5. Indirana phrynoderma (WG) 6. Terorana khare (NE) 7. Rhacophorus tuberculatus (NE) 8. Ingerana charlesdarwini (AN) 9. Bufo hololius (Penninsular) 10. Microhyla sholigari (Penninsular) 11. Bufoides meghalayanus (NE) 12. Rhacopharous pseudomalabaricus (WG) 13. Pedostibes kempii (NE) 14. Pedostibes tuberculosus (WG) 15. Polypedates insularis (AN) 16. Nyctibatrachus vasanthi (WG) 17. Raorchestes chalazodes (WG)

* This list will be populated with more species as better assessment of the status of various species is made available.

Zoological Park and their practice species 1. Arignar Anna Zoological Park, Vandalur, Chennai a) Eyphylctis cyanophylctis b) Duttaphrynus melanostictus c) Ramanella variegata d) Polypedates maculates

2. Nandankanan Biological Park, Bhubaneshawar a) Sphaerotheca breviceps b) Polypedates maculatus c) Duttaphrynus melanostictus d) Kalula pulchra

3. Madras Crocodile Bank Trust, Mamallapuram a) Duttaphrynus melanostictus b) Sphaerotheca rolandae c) Polypedates maculatus d) Ramanella variegata

4. Chennai Snake Park, Chennai a) Uperodon systoma (All India) b) Fejarvarya limnocharis c) Duttaphrynus melanostictus

691 Ex situ Management of Amphibians

5. Rajiv Gandhi Zoological Park, Pune a) Microhyla ornata b) Duttaphrynus melanostictus c) Polypedates maculatus

6. Padmaja Naidu Himalayan Zoological Park, Darjeeling a) Tylototriton verrucosus b) Duttaphrynus himlayanus c) Ichthyophis sikkimensis

7. Pilikula BiologicalPark, Mangalore a) Hylarana malabarica b) Indirana brachytarsus c) Ramanella montana d) carnosus

8. Biological Park, Itanagar a) Polypedates leucomystax b) c) Leptobrachium smithi d) Rhacophorus maximus

9. Nehru Zoological Park, Hyderabad a) Duttaphrynus melnostictus b) Sphaerotheca breviceps c) Polypedates maculates

10. Sri Chamarajendra Zoological Gardens, Mysore a) Hylarana malabarica b) Clinotarsus curtipes c) Microhyla rubra d) Ichthyophis beddomi

11. V.J.B. Udyan Zoo, Byculla Mumbai a) Raorchestes bombayensis b) Sphaerotheca breviceps c) Indirana leithii

12. Bahinabhai Prani Sangrahalaya, Pimpri, Pune a) Rhacophorus malabaricus b) Raorchestes bombayensis c) Hylarana malabarica

13. National Zoological Park, Delhi a) Fejervarya limnocharis b) Bufo stomaticus c) Polypedates maculatus

14. M.C. Zoological Park, Chhatbir, Chandigarh a) Chirixalus dudhwaensis b) Fejervarya limnocharis c) Bufo stomaticus

701 Ex situ Management of Amphibians

15. Tata Steel Zoological Park, Jamshedpur a) Bufo stomaticus b) Uperodon systoma c) Polypedates maculatus d) Sphaerotheca dobsoni

16. Bhagwan Birsa Zoological Park, Ranchi a) Bufo stomaticus b) Uperodon systoma c) Polypedates maculatus d) Sphaerotheca dobsoni

17. Pt. Govind Ballabh Pant High Altitude Zoo, Nainital a) Duttaphrynus himalayanus b) Fejervarya limnocharis c) Paa vicina

18. Assam State Zoo & Botanical Garden, Guwahati a) Chirixalus simus b) Kaloula pulchra c) Fejervarya limnocharis d) Bufo stomaticus e) Polypedates leucomystax

19. Seppaijala Zoological Park, Sepahijala a) Kaloula pulchra b) Fejervarya limnocharis c) Bufo stomaticus d) Polypedates leucomystax

20. Aizwal Zoo, Aizwal a) Kaloula pulchra b) Fejervarya limnocharis c) Bufo stomatica d) Rhacophorus maximus

21. Manipur Zoological Park, Imphal a) Occidozyga borealis b) Bufo stomaticus c) Polypedates leucomystax

22. Biological Park, Chidiyatapu, Andaman & Nicobar Islands a) Ingerana charlesdarwini b) Microhyla inornata

23. Laboratory for Conservation of Endangered Species (LaCONES), Hyderabad a) Duttaphrynus melanostictus b) Sphaerotheca breviceps c) Polypedates maculates

WL- Western Ghats NE- North East H- Himalayan AN – Andaman & Nicobar Islands 711 Notes About the Authors

Brij Kishor Gupta is an Evaluation & Monitoring Officer with Central Zoo Authority (CZA) since 2000. He is also member of IUCN/SSC/Conservation Breeding Specialist Group & South Asian Amphibian and Reptile Specialist Group as well as Country Representative for Durrell Conservation Academy, United Kingdom.

Benjamin Tapley is a Team Leader, Herpetology Department, Zoological Society of London, London Zoo, London, United Kingdom.

Karthikeyan Vasudevan is a Senior Principal Scientist at Laboratory for Conservation of Endangered Species (LaCONES), Centre for Cellular & Molecular Biology, Hyderabad, India.

Matt Goetz is a Head of Herpetology Department, Durrell Wildlife Conservation Trust, Jersey, United Kingdom. c/o IUCN SSC Conservation Breeding Specialist Group 12101 Johnny Cake Ridge Rd Apple Valley, MN 55124 USA www.AmphibianArk.org

Message Securing habitat for threatened species is necessary, but in some instances not sufficient to insure the survival of the species. Polluon, disease, invasive species and other factors within the habitat must be migated, and oen this cannot happen in me to prevent species exncons.In such cases the development of capve populaons may be the only hope.

With the threat of amphibian exncons looming large, the ability to maintain and reproduce amphibians in zoos and aquariums is becoming increasingly important. The Amphibian Conservaon Acon Plan (ACAP) (Gascon et al., 2007), a product of the Internaonal Union for the Conservaon of Nature Species Survival Commission Amphibian Conservaon Summit in 2005, specifically idenfies the need for capve programs, “In the face of overwhelming and somemes urgent threats to many amphibians, such as disease or habitat destrucon, the only hope in the short‐term for populaons and species at immediate risk of exncon is immediate rescue for the establishment and management of capve survival‐assurance colonies.”.

Drawing on published literature and on the extensive experience of the editors, this volume provides invaluable informaon relevant to the establishment of those capve colonies called for in the ACAP. While not all of the species for which husbandry protocols are described are threatened at present, many are species for which populaons are declining and which may soon be at risk. Addionally, some may serve as models for more endangered species, allowing the development of husbandry skills necessary for working with more threatened taxa.

Assam State Zoo, Guwaha and Central Zoo Authority is to be commended for developing this volume, which will undoubtedly become a much‐used reference for all zoos seeking to enhance their ability to care for and conserve amphibians in the South Asia region.

Anne Baker Execuve Director

September 27, 2015

Amphibian Ark is a partnership among the World Association of Zoos and Aquariums, the IUCN SSC Conservation Breeding Specialist Group, and the IUCN SSC Amphibian Survival Alliance.