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Journal of Threatened Taxa Building evidence for conservaton globally www.threatenedtaxa.org ISSN 0974-7907 (Online) | ISSN 0974-7893 (Print) Communication Amphibian abnormalities and threats in pristine ecosystems in Sri Lanka

G.K.V.P.T. Silva, W.A.D. Mahaulpatha & Anslem de Silva

26 December 2019 | Vol. 11 | No. 15 | Pages: 15004–15014 DOI: 10.11609/jot.5394.11.15.15004-15014

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Threatened Taxa Journal of Threatened Taxa | www.threatenedtaxa.org | 26 December 2019 | 11(15): 15004–15014

Amphibian abnormalities and threats in pristine ecosystems

Communication in Sri Lanka

ISSN 0974-7907 (Online) 1 2 3 ISSN 0974-7893 (Print) G.K.V.P.T. Silva , W.A.D. Mahaulpatha & Anslem de Silva

PLATINUM 1, 2 Department of Zoology, Faculty of Applied Sciences, University of Sri Jayewardenepura Gangodawila, Nugegoda, OPEN ACCESS Sri Lanka. 315/1, Dolosbage Road, Gampola (Central Province), Sri Lanka. 1 [email protected], 2 [email protected] (corresponding author), 3 [email protected] (corresponding author)

Abstract: Amphibian abnormalites are caused by numerous etologies prevailing in the environment. Since amphibians are good bio indicators of the environment, amphibian abnormalites are popularly known as a veritable ecological screening tool to assess ecosystem health. The present study was carried out encompassing within and outside the Horton Plains Natonal Park areas, from January to November 2017. Distributon of amphibian morphological abnormalites were assessed in and around the fve lentc water bodies through gross visual encounter. Six quadrates of 1m×2m were randomly placed in each sampling site. Frequency and compositon of amphibian abnormalites were assessed in a total of 694 amphibians, belonging to four families and 11 species. Thereby, 4.5% and 80.87% abnormality indexes were accounted for respectvely within and outside the park, comprehended surfcial abnormalites, ectromelia and femoral projecton abnormality types. Surfcial abnormalites were the most predominant in both localites, generally occurring at the hind limb region of pre-mature stages of Taruga eques. Two lentc water bodies were identfed as “abnormality hotspots” within and outside the Horton Plains Natonal park; however, a multplicity of possible combinatons of potental causes of abnormalites were present in the environment. Hence, fnding the exact causes of amphibian abnormalites are an extremely difcult exercise in the feld.

Keywords: Abnormality, Horton Plains Natonal Park, morphological, Taruga eques.

DOI: htps://doi.org/10.11609/jot.5394.11.15.15004-15014

Editor: Neelesh Dahanukar, Indian Insttutes of Science Educaton and Research (IISER), Pune, India. Date of publicaton: 26 December 2019 (online & print)

Manuscript details: #5394 | Received 08 January 2019 | Final received 06 November 2019 | Finally accepted 07 December 2019

Citaton: Silva, G.K.V.P.T., W.A.D. Mahaulpatha & A. de Silva (2019). Amphibian abnormalites and threats in pristne ecosystems in Sri Lanka. Journal of Threatened Taxa 11(15): 15004–15014. htps://doi.org/10.11609/jot.5394.11.15.15004-15014

Copyright: © Silva et al. 2019. Creatve Commons Atributon 4.0 Internatonal License. JoTT allows unrestricted use, reproducton, and distributon of this artcle in any medium by adequate credit to the author(s) and the source of publicaton.

Funding: University of Sri Jayewardenepura.

Competng interests: The authors declare no competng interests.

Author details: G.K.V.P.T Silva is a graduate student from the University of Sri Jayewardenepura with BSc (Special) in Zoology and presently working as the department research assistant on wildlife management and conservaton engaged with the “Wildlife Circle”, Department of Zoology, University of Sri Jayewardenepura. Prof. W.A.D. Mahaulpatha is a researcher and a professor of zoology at the University of Sri Jayewardenepura. She has undertaken many pieces of research in wildlife conservaton and management including ornithology, herpetology and mammalogy, and she has more than a hundred publicatons. As result she received the Presidental Award in 2018. Anslem de Silva MSc. DSc. has contributed approximately 425 papers. This includes about 50 books and the latest book was “Naturalist Guide to Reptles of Sri Lanka’ (2017) published in the United Kingdom. He has received the Presidental Award for scientfc publicaton four tmes. He is currently working as regional chairman of the Crocodile Specialist Group IUCN for South Asia and Iran.

Author contributon: All authors contributed equally. GKVPTS was the main researcher on this research project, feld sampling, data collecton, data analysis and preparaton of the paper. WADM was the supervisor of this piece of research and provided immense guidance and support from the initaton of the project untl the completon, with her valuable experience and knowledge. AdS provided some important relevant literature and valuable suggestons with his expertse in feld studies, which contributed a lot to improve parts in the methodology secton and to prepare the manuscript.

Acknowledgements: I would like to thank the Department of Wildlife Conservaton for providing me the permission to conduct this research within and outside the Horton Plains Natonal Park (WL/3/2/28/17) and staf of Horton Plains Natonal Park for the received immense support. I would like to extend my grattude to Prof. (Mr.) M. Lannoo, Prof. (Mrs.) R. Rajakaruna and Dr. (Mr.) K. Ukuwela for their comments and valuable suggestons. I greatly appreciate the support that I received from all the members of wildlife circle of the University of Sri Jayewardenepura, Department of Zoology.

15004 Amphibian abnormalites and threats in pristne ecosystems in Sri Lanka Silva et al.

INTRODUCTION deformed amphibians (Gillilland & Muzzall 1999). There is a considerable efect on anuran abnormalites “Abnormality” refers to “any deviaton from normal from predators existng in natural environments morphology, independent of whether its origin was (Johnson & Bowerman 2010). Most of these amphibian developmental or acquired afer proper development” abnormalites are caused by aquatc predators such as (Lunde & Johnson 2012). Both amphibian malformatons dragonfy larvae (Bowerman & Johnson 2010; Johnson and deformites are included in amphibian abnormalites & Bowerman 2010), small fshes, crabs, crayfshes (Reeves et al. 2008). Amphibian abnormalites can be (Johnson et al. 2001a, 2006), diving beetles, predatory classifed mainly as surfcial abnormalites (infectous odonate nymphs and water scorpions (Ballengee & diseases/cysts and wounds), skeletal abnormalites and Sessions 2009). Smaller aquatc predators (insect larvae, eye abnormalites (Linder 2003; Reeves et al. 2008). small fshes) atack the exposed portons of larval and Amphibian abnormalites have interconnected with metamorphosis stages of anurans such as the tail or many factors including, chemical contaminants (Bridges limbs (Bowerman & Johnson 2010). A traumatc loss of et al. 2004; Lunde & Johnson 2012), trematode, cestode, an entre limb (Lannoo 2008), however, and wounds of and nematode parasites (Ankley et al. 2004; Imasuen & amphibians can be seen afer the metamorphosis which Ozemoka 2012), predators (Johnson et al. 2006; Johnson are produced by the atack of large vertebrate predators & Bowerman 2010) and UV (Ultraviolet) radiaton (Bowerman & Johnson 2010). (Blaustein & Johnson 2003; Lannoo 2008). Leech atack causes some of the abnormalites of Anthropogenic actvites have been recognized as a amphibians and many abnormality studies have proved main element for the modifcaton of aquatc habitats that leech atacks cause a high prevalence of missing (Johnson & Chase 2004). Especially nutrient loading in limbs (ectromelia) or parts of the limbs (Johnson et freshwater ecosystems (partcularly ponds) directs to al. 2001a, 2002, 2006; Ballengee & Sessions 2009; the acceleraton of eutrophicaton that result in shifing Bowerman & Johnson 2010). the community compositon. This results in various Considering worldwide abnormality studies it can be amphibian abnormalites. seen that most of them have been carried out as laboratory Helminth infectons are considered as a major studies, related with chemical inductons (Burkhart et al. governing factor for existng amphibian abnormalites 1998; Lajmanovich et al. 2003) and parasitc inductons and linked with numerous factors. Most of the trematode (Johnson et al. 2001a; Stopper et al. 2002) to frog infectons in amphibians are recorded from the habitats embryos and tadpoles in various limb bud stages. Only a associated with agriculture and catle farms. Trematodes few studies, however, have been conducted to investgate in the genus Ribeiroria occur in lentc aquatc water bodies abnormalites of amphibians in the feld (Johnson et (Johnson et al. 2004). Aquatc snails in lentc aquatc al. 2001b; Peltzer et al. 2011). As regards the fact that water bodies serve as intermediate host for Ribeiroria Sri Lanka is a biodiversity hotspot, only four studies parasite. When high nutrient levels are present the have been mentoned (Rajakaruna & Samarawickrama snail densites increase exponentally. Free swimming 2007; de Silva 2009, 2011; Meegaskumbura et al. 2011). cercariae emerge from infected snails then penetrate and Though, abnormality surveys were based on selected encyst as metacercariae in the second intermediate host regions of the country, it’s more valuable to extract data (Jayawardena et al. 2010b) ofen around the developing from pristne ecosystems with the purpose of identfying limb buds of amphibian larvae, leading to improper limb the actual threats of amphibian’s survival. Therefore, development (Blaustein & Johnson 2003). In additon the present research was undertaken to assess the to that frequent exposure of amphibians to chemical amphibian abnormalites and their possible predators contaminants accumulated in lentc water bodies leads which cause amphibian abnormalites encircling the to a reducton of immunity strength. As a result of area in and around the lentc water bodies within and that amphibians become highly susceptble to parasitc outside the World Heritage Horton Plains Natonal infectons (Kiesecker 2002; Budischak & Belden 2009; Park as a comparison of threats and abnormality types Lunde & Johnson 2012). Cestodes are one of the major possessed by amphibians against two diferent localites parasite groups which infect the amphibians. Cestodes to ultmately fulfll the knowledge gap of feld studies commonly occur within the musculature of the body and pertnent to amphibian abnormalites. in the hind limbs of adult and juvenile frogs (Gillilland & Muzzall 2002) and a number of metacercariae of helminthes has been recorded in the leg musculature of

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MATERIALS AND METHODS 1A). Sampling sites, outside the HPNP were located within a range of 1,170.9–1,864.7 m elevaton and mainly a) Study site associated with forested area. GPS (Global positoning The study was conducted within and outside the system) points of these sampling sites were recorded Horton Plains Natonal Park (HPNP) from January to using Garmin etrex Euro hand held GPS receiver. November 2017. HPNP is located between 6.802 Six quadrates each 1 x 2 m were placed within each northern lattudes and 80.807 eastern longitudes sampling site for the sampling of amphibians. Three (Green 1990) which occupies an area of 3,160ha and is quadrates were randomly placed in the area 1m from contguous with the Peak Wilderness Sanctuary to the the pond bank outside the pond and three quadrates west. It is in the eastern extremity of the Nuwara Eliya were randomly placed inside the pond in the area of 1m District in Sri Lanka within the range of 2,100-2,300 m from the pond bank (Faruk et al. 2013). Anuran species elevaton (DWC 2007). Tropical montane cloud forests (larvae, metamorphs, juveniles, and adults) and leeches and wet pathana grasslands are the two distnct habitats in these quadrates were surveyed from morning to afer in the park (Gunatlleke & Gunatlleke 1990) with a noon (08.00–16.00 h) and during the night (18.00–20.00 narrow ecotone belt of shrubs and herbs between the h). Moreover, vegetaton within each plot was searched two. even when slight movement was detected. Head lamps Most of the lentc water bodies are surrounded and torches were used for nocturnal searches. Larval by three main grass types (Chrysopogon nodulibarbis, amphibians were captured using actve sweeps (Dodd Andropogon polyptychos, and Garnota exaristata) 2010), metamorphic anurans were captured with and one bamboo species (Arundinaria densifolia) in a dip net or by hand (Ouellet et al. 1997) and adult the grassland habitat (DWC 2007). The sampling was amphibians were captured with a dip net or by hand conducted in selected fve lentc water bodies on four (Wheeler & Whelsh 2008; Urbina & Jenny 2009). A days per month, providing equal efort to each sampling small amount of water was added to the container to site. Three lentc water bodies (A, B and C) were selected prevent overheatng and desiccaton (Wheeler & Whelsh within the HPNP and two (D and E) were selected outside 2008). Overcrowding based on the container size and the HPNP based on the availability of amphibians (Figure Tamb was avoided (Lunde & Johnson 2012). At the end

Figure 1. (A). Locatons of sampling sites within and outside the Horton Plains Natonal Park. [Lentc water bodies within the HPNP]. A (6.8390N and 80.8120E), B (6.8340N and 80.8090E), C (6.8010N and 80.8070E), D (6.8500N and 80.8180E), E (6.8050N and 80.9080E).

15006 Journal of Threatened Taxa | www.threatenedtaxa.org | 26 December 2019 | 11(15): 15004–15014 Amphibian abnormalites and threats in pristne ecosystems in Sri Lanka Silva et al. of the survey they were released back to their habitat body. Amphibian invertebrate predators in the ponds afer removing the ectoparasites from the atached skin were classifed as either abundant (<100 individuals surface of the amphibians. Amphibian species and their trap per day) or rare (<10 individuals trap per day) based stages were identfed using the amphibian guides of on density capture per day according to Bowerman & Duta & Manamendra-Arachchi (1996), Manamendra- Johnson (2010). Possible large vertebrate predators Arachchi & Pethiyagoda (2006), and de Silva (2007). (aquatc birds) of amphibians were assessed based on When an amphibian was captured, (a) it was identfed to observatons in and around the lentc water bodies. species level, (b) life stage noted, (c) presence or absence Aquatc invertebrate predators of amphibians were of the abnormalites assessed (in larvae, metamorphs, identfed based on the morphological characteristcs juvenile and adult stages) (If abnormalites were present, (nature of the abdomen, presence of exoskeleton and they were recorded with the count, according to the arrangement of gills) with the help of Curts (2011) abnormality classifcatons of guides) and recorded. and Quek et al. (2014). Possible large vertebrate predators (aquatc birds) of amphibians were identfed b) Identfcaton of amphibian abnormalites (colouraton of feathers and beak and foot type) using Amphibian physical abnormalites (assessed the Harrison (2011), a bird guide. All types of abnormalites, abnormalites which appeared externally) were aquatc invertebrates and large vertebrate predators as associated with the loss of body parts or organs. possible threats of amphibians (which cause amphibian External abnormalites were identfed through gross abnormalites) were photographed using a digital camera. visual inspectons (both dorsal and ventral sides of the amphibian body) and classifed based on Meteyer (2000), d) Determinaton of the environmental variables Johnson et al. (2001b), Johnson et al. (2002), Lannoo Environmental variables of lentc water bodies (2008), Rajakaruna et al. (2008), Reeves et al. (2008), were measured to fnd out the habitat suitability for Johnson & Hartson (2009), and Peltzer et al. (2011) amphibians (Hamer & Lane 2002; Urbina & Jenny guides. Surfcial abnormalites (cysts) were identfed by 2009; Dodd 2010; Sparling 2010). The atmospheric careful examinaton of the external body surface (dorsal data temperature (ºC) and relatve humidity (RH) were side, ventral side, around the fore limbs and hind limbs) assessed. Ambient temperature (Tamb) was recorded and using the external morphological features of the 2m above the water surface / ground (using Kestrel 4000 infecton. Cysts of infected amphibians are found as clear weather meter, USA) and relatve humidity (RH) data were or brownish colour (Johnson et al. 2004), round, swelling recorded 2m above the ground. In additon to that soil nodules of the musculature (Ostler 2004). Moreover, pH values were obtained using Kelwey soil tester, under Johnson et al. (2004), Ostler (2004), de Silva (2007, the soil chemical data. Furthermore, dissolved oxygen 2009), and Jayawardena et al. (2010a, b) were referred (DO) (YSΙ 550A Dissolved Oxygen Instrument), water to for the identfcaton of cysts. Based on the number temperature (Tw), pH (YSΙ Eco Sense pH 100A meter), and of cysts (surfcial), severity of amphibian infectons conductvity (YSΙ Eco Sense EC300A Conductvity meter) were classifed as mild and moderate. Mild infectous were measured using analysis of water physiochemical amphibians possess 1–3 nodules and moderate infectous data. Soil chemical data and water physiochemical data amphibians possess 4–6 nodules (de Silva 2009). were recorded once a month in each plot (as mentoned above) using a standardized data sheet. c) Identfcaton and sampling of possible predators of amphibians e) Data analysis Leeches were identfed by careful examinaton of the The Minitab version 17.0 statstcal sofware was external body surface of the anuran larvae, metamorphs, used to calculate the mean and standard deviaton juveniles, adults and also the grasses and shrubs which (SD) values of environmental variables and possible were adjacent to the bank of the water body. Possible predators of amphibians in each sampling site. Graphical aquatc invertebrate predators of the amphibian life representatons were created using Microsof Excel 2013 stages (larvae, metamorphs, juveniles and adult) were sofware. Abnormality index (AI) was calculated using sampled by using the 0.36 × 0.22 m dip net, performing the equaton, 2m sweeps at 11 evenly spaced points around each AI = (Total number of abnormal amphibians / Total pond’s perimeter (Bowerman & Johnson 2010). number of amphibians inspected) × 100% Captured aquatc predatory varietes were identfed, counted, recorded and released to the same lentc water

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RESULTS both localites. Abnormal metamorphs were found only inside the HPNP which accounted for 4.35% of the a) Abundance of amphibian species abnormalites. A high AI, however, was observed in the A total of 694 amphibians belonging to four families juvenile stage than in adults in both localites—56.52% and 11 species were recorded during the study in the fve and 77.70% within and outside the HPNP. Moreover, lentc water bodies studied. Five-hundred-and-eleven most of the abnormalites were found at the hind limb individuals were examined inside the HPNP and 183 region of amphibians (Figure 2D). When comparing the individuals were examined outside the HPNP (Table 1). AI of amphibians in each sampling site, no abnormal amphibians were recorded in pond C. Amphibians b) Amphibian abnormalites with ectromelia were only recorded within the HPNP Out of the 511 amphibians examined inside the HPNP, in sampling site B; however, 1.16%, 14.39%, 100%, and only 23 (4.50%) had abnormalites; however, 80.87% of 2.78% abnormality percentages were recorded in A, B, amphibians outside the HPNP, possessed abnormalites. D, and E ponds, respectvely (Table 2). With reference Taruga eques had the most number of abnormalites to natonal conservaton states (determined by IUCN), in both localites—22 inside the HPNP and 147 outside 94.71% of the amphibians found in the HPNP were the HPNP. This was 99.30% of AI outside the HPNP and endangered, thereby 4.55% had abnormalites, whereas 95.70% of AI inside the HPNP with respect to T. eques. in contrast 100% of the amphibians were abnormal Surfcial abnormalites (Image 1) were more dominant outside the HPNP. than ectromelia and femoral projecton abnormality types in both localites, thereby mild infectons (60.00%) c) Comparison of amphibian predatory density in each (Image 1A and 2B) and moderate infectons (77.55%) sampling site (Image 1C) were predominant respectvely within and Damselfy larvae Elatoneura leucostgma were the outside the HPNP (Figure 2A). Cysts were recorded only most abundant possible predatory type of the amphibians. in T. eques, accounted for 5.76% within the HPNP and Damselfy larvae (Image 3A) were recorded in A, B and C 100.00% outside the HPNP (Figure 2B). Ectromelia was sampling sites within the HPNP. Water scorpions (Image recorded in both T. eques (Image 2A) (0.58%) and M. 3C), dragonfy larvae Orthetrum glaucum (Image 3B), greenii (Image 2B) (0.74%) species. Femoral projecton Pond Herons Ardeola grayii (Image 3E) were recorded was found only in F. cf. limnocharis (Image 2C) which only in pond A. Leeches (Image 3D) were recorded in A accounted for 5.56% frequency of the abnormalites. and B sampling sites. Crabs Perbrinckia glabra (Image 3D) Comparing the abnormality index with amphibian life were only found in sampling site C. No possible predators stages (Figure 2C), no abnormal larvae were found in of amphibians were recorded in pond E. Aquatc beetles were only recorded in pond D (Table 3).

Table 1. Amphibian abundance in and around the lentc water bodies d) Environmental variables of sampling sites within and outside the Horton Plains Natonal Park. Highest conductvity was recorded in sampling site E. It may due to the fact that the water was contaminated Locality Abnormality type Family & Species Abundance Index (%) with agro-chemicals. Moreover, D lentc water body Dicroglossidae recorded the least water pH value which indicates acidic Minervarya greenii 135 0.74 conditons and that more anthropogenic stressors are Microhylidae Microhyla zeylanica 11 00 present in the water body (Table 4). Microhylidae HPNP Uperodon palmatus 03 00 Rhacophoridae Pseudophilautus alto 02 00 DISCUSSION Pseudophilautus frankenbergi 01 00 Pseudophilautus microtypanum 12 00 Taruga eques 347 6.34 Both amphibian malformatons and deformites Bufonidae are included in amphibian abnormalites (Reeves et al. Dutaphrynus melanostctus 06 00 2008). Some of the abnormalites are external physical OHPNP Dicroglossidae Euphlycts cyanophlycts 04 00 abnormalites and others are internal (Blaustein & Minervarya kirtsinghei 08 00 Johnson 2003; Spolyarich et al. 2011). The present study Fejervarya cf. limnocharis 18 5.55 Rhacophoridae did not fnd any abnormalites in the family Bufonidae Taruga eques 147 100 coinciding with the studies of Lannoo (2008) who

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B C Image 1. A and B—mild infecton: infected Taruga eques amphibians with clear colored 1–3 cysts (surfcial abnormalites) as round swelling nodules of the musculature and subcutaneous tssue of the skin | C—moderate infecton- Infected Taruga eques amphibians with 4–6 cysts. © G.K.V.P.T. Silva.

A C

B D

Image 2. A—Ectromelia in Taruga eques | B—Ectromelia in Minervarya greenii metamorph | C—femoral projecton abnormality in Fejervarya cf. limnocharis | D—leeches atached to the external body surface of Taruga eques. © G.K.V.P.T. Silva.

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Table 2. Diferent abnormality types and abnormality indices recorded in each sampling site.

Locality type Sampling site Ectromelia Infectons Femoral projectons Abnormality index (%) A - 4 - 01.16 HPNP B 3 16 - 14.39 C - - - 00.00 D - 147 - 100.00 OHPNP E - - 1 02.78

Table 3. Comparison of density of amphibian possible predators in each sampling site number of predators trapped/observed per day.

Locality type

HPNP OHPNP

Predator A B C D E

1.89 ±0.93 - - - - Aquatc beetles Crustaceans - 1.44±1.01 - - -

Damselfy larvae 1 .78±1.92 104.33±31.89 1.22±1.09 - -

Dragonfy larvae 11.56±6.73 - - - -

Leeches 01.56±2.01 5.44±2.74

Pond herons 02±0.87 - - - -

Water scorpions 1 .56±1.01 - - - -

Aquatc beetles - - - 01±1.12 -

Table 4. Environmental variables in and around the lentc water bodies in each sampling site.

Environmental variables (Mean ± SD) Site DO Conductvity T (˚C) RH Soil pH T (˚C) Water pH amb (mg/L) w (µS/cm) A 16.29 ± 0.30 94.11±2.57 5.90±0.38 4.67±0.19 15.73±0.53 8.14±0.68 18.56±5.68

B 16.15±0.58 95.05±2.06 5.91±0.50 4.55±0.26 15.75±0.45 7.93±0.62 17.07±5.79

C 15.88±0.32 94.50±2.77 5.88±0.45 4.65±0.23 16.01±0.66 8.30±0.66 20.95±7.13

D 16.80±0.67 95.03±2.66 4.49±0.18 4.52±0.16 16.21±0.77 6.89±0.90 18.94±4.03

E 15.41±0.89 91.95±8.14 5.15±0.45 4.66±0.19 16.99±0.38 6.95±0.93 25.65±4.50

observed that the Bufonidae family is less susceptble to to abnormalites (Johnson & Bowerman 2010). B lentc abnormalites. water body was located within the pristne ecosystem of Previous studies have indicated that less than 5% HPNP. Moreover, all the water quality parameters were abnormality prevalence of the populaton in a partcular also within the standard levels for amphibian survival. area or site is normal (Johnson et al. 2002; Kiesecker Therefore, further experiments must be carried out to et al. 2004; Piha & Pekkonen 2006; Lunde & Johnson fnd out the exact cause for abnormalites in B water 2012). The abnormality prevalence within the HPNP body. In contrast, D lentc water body was located on was 4.50%, which is within the accepted range; however, the way to HPNP and most visitors use it as a garbage abnormality prevalence outside the HPNP was 86.55% dumping site (which may be the reason for reportng and should be considered as abnormally high (Piha & less DO and soil pH values). As a result of that, this high Pekkonen 2006). Lentc body wise, pond B inside the nutrient content of the water body provides a beter HPNP had 14.5% abnormality prevalence and pond D had environment to increase the parasitc density. It might abnormality prevalence of 100%. These high abnormality be the major reason for recording 100% of surfcial prevalent sites were classifed as ‘‘hotspots’’ with respect abnormalites in D lentc water body. Since, observed

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Figure 5. A—comparison of the infecton severity within and outside the Horton Plains Natonal Park (HPNP) | B—comparison of abnormality types recorded in diferent amphibian species found within and outside the HPNP | C—comparison of Percentages of abnormality indices recorded from diferent amphibian life stages within and outside the HPNP | D—comparison of abnormalites based on the body region where the abnormalites were recorded. abnormality prevalence of B and D lentc water bodies Johnson et al. 2002; Piha & Pekkonen 2006), which may were greater than the expected baseline range, further be resulted by the atack of natural pradators when they investgatons are warranted to fnd out the locality try to avoid the escape of amphibians. Results of the specifc causes (Lunde & Johnson 2012). present study also tallies with these fndings as most Highest number of amphibian abnormalites were of the abnormalites found as cysts arise within the reported in pre-matured stages of amphibians which musculature found in the hind limbs. may for the reason that either adult populatons of Surfcial abnormalites (cysts) were the predominant amphibians are less susceptble for abnormalites against abnormality type recorded in this study. Even the diferent environmental stressors or they have reduced HPNP comprises optmal conditons for growth of survivorship. The survival of abnormal amphibians Batrachochytrium dendrobatdis, in the most observed declines due to a high predaton pressure and inability cysts may be caused by parasitc helminths infected afer to capture its prey (Lunde & Johnson 2012). Since limb bud stages of amphibians since they were located abnormal adult amphibians are unable to survive long within musculature, mostly around limb structures and periods of tme in the environment, frequently a smaller were easily visible to the naked eye as swelling round number of adult amphibian populatons are discovered nodules; in contrast pathogenic cysts are microscopic and with abnormalites (Goodman & Johnson 2011; Lunde confned only to the epidermis (Lunde & Johnson 2012). & Johnson, 2012). Previous abnormality studies have Lunde & Johnson (2012) also recorded that the high also observed that most of the amphibian abnormalites number of amphibians inhabitng lentc water bodies are are associated with hind limbs (Ouellet et al. 1997; commonly infected with many trematode species that

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Lunde & Johnson, 2012). These two factors may have contributed to amphibian abnormalites in the lentc water bodies that were studied. All the amphibian abnormalites cannot be explained away as parasitc infectons. Lannoo (2008) and Lunde & Johnson (2012) observed that high level of ectromelia

A B are present in frogs even when parasites are absent. In additon to parasites, amphibian predators (vertebrates and invertebrates) play a major role in limb abnormalites in amphibians (Bowerman & Johnson 2010). Larval amphibians are atacked by small predators including aquatc invertebrates; however, traumatc loss and injuries in limbs of amphibians (afer metamorphosis) are mostly caused as a result of failed predatory atempts by large vertebrate predators (Bowerman & Johnson 2010). Bowerman & Johnson (2010) observed C a direct relatonship between the leech density and the abnormality level in feld studies and laboratory inducton of leeches. Leeches act as predators at amphibian larval stages and act as parasites of both larval and adult stages E (Schalk & Forbes 2002; McCallum et al. 2011). Damselfy larvae was the abundant possible predators recorded in D E site B and all the ectromelia were observed in the same Image 3. Possible predators of amphibians which cause amphibian pond. Therefore, there is a high possibility of these abnormalites. A—Damselfy larva | B—Dr agonfy larva | C—Water predators causing the ectromelia observed in site B. scorpion | D—Crustacean (crab) | E—Pond Heron. © G.K.V.P.T. Silva. Water quality values of all sampling sites were included to the estmated optmum ranges (Sparling form cysts under the skin within musculature. 2010) for amphibian growth and development; however, Amphibian abnormalites due to trematode with respect to soil pH values, even moderate acidic infectons depend on the stage of limb development at conditon is identcal to HPNP (Chandrajith et al. 2009), which infectons occurred (Schothoefer et al. 2003). strongly acidic conditons can’t be expected within the Therefore, tming of the infecton is a critcal determinant sampling sites outside the HPNP, which indicate that in forming abnormalites (Jayawardena et al. 2010b). both sampling sites outside the HPNP may have exposed Infecton coincides at pre-limb bud developmental stage to some anthropogenic actvites. of the tadpoles which results in high mortality rate with Multplicity of possible combinatons of potental axial abnormalites. Trematode infectons acquired causes of abnormalites are present in the environment. at limb bud stage also produces high abnormality Hence, fnding the exact causes of amphibian rate including ectromelia (Schothoefer et al. 2003; abnormalites are an extremely difcult exercise in the Jayawardena et al. 2010b). Further more, there is no feld. The fact that abnormalites are linked with ecology, any efect to limb development and survival of tadpoles epidemiology, and developmental biology further when they are infected at the paddle stage (Schothoefer increases the complexity of this problem. et al. 2003; Jayawardena et al. 2010b). However afer the infecton at the paddle stage of amphibians, encysted parasites are able to remain viable even at the adult CONCLUSIONS stage of amphibians (Imasuen & Ozemoka 2012). This was also observed in the amphibians in the present Taruga eques was the most susceptble amphibian study. Existence of amphibian predators in ponds species to abnormalites irrespectve of the locality type, generally increase the trematode infecton of amphibians indicated that amphibians may have been exposed to (Thiemann & Wassersug 2000; Lunde & Johnson, 2012). some stress conditons linked with multple factors. Trematodes commonly occur in the lentc water bodies Present research fndings revealed that toads are less with dragonfy larvae (Bowerman & Johnson 2010; susceptble to amphibian abnormalites. Since, B and

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D lentc water bodies were discovered as abnormality Environmental Health Perspectves 106(12): 841. htps:// hotspots, further experiments are crucial at those dx.doi.org/10.1289%2Fehp.106-1533234 Chandrajith, R., N. Koralegedara, K.B. Ranawana, H.J. Tobschall & C.B. partcular lentc water bodies, for determining the site- Dissanayake (2009). Major and trace elements in plants and soils specifc cause for amphibian abnormalites. The present in Horton Plains Natonal Park, Sri Lanka: an approach to explain forest die back. Environmental Geology 57(1): 17–28. htps://doi. study clearly discloses further that Sri Lankan tree frogs org/10.1007/s00254-008-1278-0 are in a critcal state. Since Taruga eques is confned only Curts, S. (2011). Water Bugs and Water Scorpions (Fact sheet). South to the central hill regions of the country with declining Brisbane: Queensland Museum, 487pp. de Silva, A. (2007). Amphibians of Sri Lanka: A Photographic Guide to populaton and all the amphibians which belong to Common Frogs, Toads and Caecilians. Kandy: Privately published. endangered conservaton state had abnormalites outside de Silva, A. (2009). The incidence and patern of malformatons, the HPNP, it is clear that mandatng urgent measures to abnormalites, injuries, and parasitc infecton of amphibians in Sri Lanka (preliminary fndings). Final Report: Amphibian Specialists carryout extensive research-based conservaton work Group, Seed Grant, 40. before including it to critcally endangered state is de Silva, A. (2011). Some observatons of malformaton, eye disease, needed. parasitc and viral infecton and the efects of agrochemicals on amphibians in Sri Lanka. FrogLog, 24pp. The present research data reveals that potental Dodd, C.K. (2010). Amphibian Ecology and Conservaton: A Handbook predators of amphibians are provided with excellent of Techniques. Oxford University Press, New York, 584pp. environmental conditons for their existence within the Duta, S.K. & K. Manamendra-Arachchi (1996). The Amphibian Fauna of Sri Lanka. Colombo: Wildlife Heritage Trust of Sri Lanka, 230pp. HPNP as a protected area in contrast to areas outside DWC (2007). Biodiversity Baseline Survey: Horton Plains Natonal Park. the HPNP. The present research, however, suggests Consultancy Services Report prepared by Green, M.J.B. (ed.), De Alwis, S.M.D.A.U., Dayawansa, P.N., How, R., Singhakumara, B.M.P., that exigency of extensive research-based studies for Weerakoon, D. and Wijesinghe, M.R. ARD Inc in associaton with the identfcaton of complex causes for abnormalites in Infotech IDEAS and GREENTECH Consultants. Sri Lanka Protected amphibians integrated with many disciplines, including Areas Management and Wildlife Conservaton Project (PAM &WCP/ CONSULT/02/BDBS), Department of Wildlife Conservaton, Ministry ecology, toxicology, parasitology, and developmental of Environment and Natural Resources, Colombo, 40pp. biology. These studies will be important not only as Faruk, A., D. Belabut, N. Ahmad, R.J. Knell & T.W. Garner (2013). Efects ecological or evolutonary infuences, but as signifcant of oil‐palm plantatons on diversity of tropical anurans. Conservaton Biology 27(3): 615–624. implicatons for conservaton and contemporary Gillilland, M.G. & P.M. Muzzall (1999). Helminths infectng froglets of concerns over widespread amphibian populaton losses. the Northern Leopard Frog (Rana pipiens) from Foggy Botom marsh, Michigan. Journal Helminthological Society Washington 66(12): 73– 77. htps://doi.org/10.1111/cobi.12062 Gillilland, M.G. & P.M. Muzzall (2002). 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PLATINUM The Journal of Threatened Taxa (JoTT) is dedicated to building evidence for conservaton globally by publishing peer-reviewed artcles online every month at a reasonably rapid rate at www.threatenedtaxa.org. OPEN ACCESS All artcles published in JoTT are registered under Creatve Commons Atributon 4.0 Internatonal License unless otherwise mentoned. JoTT allows allows unrestricted use, reproducton, and distributon of artcles in any medium by providing adequate credit to the author(s) and the source of publicaton.

ISSN 0974-7907 (Online) | ISSN 0974-7893 (Print)

December 2019 | Vol. 11 | No. 15 | Pages: 14927–15090 Date of Publicaton: 26 December 2019 (Online & Print) www.threatenedtaxa.org DOI: 10.11609/jot.2019.11.15.14927-15090

Peer Commentary First record in 129 years of the Tamil Treebrown Lethe drypets todara Moore, 1881 (Lepidoptera: Nymphalidae: Satyrinae) from Odisha, India by Observatons on the ex situ management of the Sumatran Rhinoceros fruit-baitng Dicerorhinus sumatrensis (Mammalia: Perissodactyla: Rhinocerotdae): – Anirban Mahata, Sudheer Kumar Jena & Sharat Kumar Palita, Pp. 15047– present status and desiderata for conservaton 15052 – Francesco Nardelli, Pp. 14927–14941 A review of the leafopper tribe Agalliini (Hemiptera: Cicadellidae: Communicatons Megophthalminae) with a revised key to the known Pakistani genera and species Revisitng genetc structure of Wild Bufaloes Bubalus arnee Kerr, 1792 – Hassan Naveed, Kamran Sohail, Waqar Islam & Yalin Zhang, Pp. 15053– (Mammalia: Artodactyla: Bovidae) in Koshi Tappu Wildlife Reserve, Nepal: 15060 an assessment for translocaton programs – Ram C. Kandel, Ram C. Poudel, Amir Sadaula, Prakrit Kandel, Kamal P. Gairhe, The windowpane oyster family Placunidae Rafnesque, 1815 with additonal Chiranjibi P. Pokheral, Siddhartha B. Bajracharya, Mukesh K. Chalise & descripton of Placuna quadrangula (Philipsson, 1788) from India Ghan Shyam Solanki, Pp. 14942–14954 – Rocktm Ramen Das, Vijay Kumar Deepak Samuel, Goutham Sambath, Pandian Krishnan, Purvaja Ramachandran & Ramesh Ramachandran, A review on status of mammals in Meghalaya, India Pp. 15061–15067 – Adrian Wansaindor Lyngdoh, Honnavalli Nagaraj Kumara, P.V. Karunakaran & Santhanakrishnan Babu, Pp. 14955–14970 Notes

A comparatve analysis of hair morphology of wild and domestc ungulate prey Recent records of the rare Mountain Tortoiseshell Aglais rizana species of Leopard Panthera pardus fusca (Mammalia: Carnivora: Felidae) (Moore, 1872) (Lepidoptera: Nymphalidae) in the upper Garhwal, from Goa, India western Himalaya, India, afer 100 years – Bipin S. Phal Desai, Avelyno H. D’Costa & S.K. Shyama, Pp. 14971–14978 – Arun Pratap Singh & Tribhuwan Singh, Pp. 15068–15071

Understanding people’s percepton and attudes towards mammalian fauna First report of Dicranocentroides indica (Handschin, 1929) (Collembola: using qualitatve data: a case study in Barail Wildlife Sanctuary, India Paronellidae) from Odisha, India – Amir Sohail Choudhury, Rofk Ahmed Barbhuiya & Parthankar Choudhury, – Ashirwad Tripathy, Pp. 15072–15073 Pp. 14979–14988 Additons to the knowledge of darkling beetles (Coleoptera: Tenebrionidae) An assessment of bird communites across Ujjani and its fve satellite wetlands from the Indo-Burma Biodiversity Hotspot, Meghalaya, India in Solapur District of Maharashtra, India – Vishwanath Datatray Hegde, Pp. 15074–15078 – Shraddha Prabhakar Karikar, Subhash Vithal Mali, Kulkarni Prasad & Aphale Prit, Pp. 14989–14997 Bhutan Asiabell Codonopsis bhutanica Ludlow (Asterales: campanulaceae): a new additon to the Indian fora Growth rate of captve Gharials Gavialis gangetcus (Gmelin, 1789) – Samiran Panday, Vikas Kumar, Sudhansu Sekhar Dash, Bipin Kumar Sinha & (Reptlia: Crocodylia: Gavialidae) in Chitwan Natonal Park, Nepal Paramjit Singh, Pp. 15079–15082 – Bed Bahadur Khadka & Ashish Bashyal, Pp. 14998–15003 Gentana urnula Harry Sm. (Gentanaceae), a new record for the fora of Amphibian abnormalites and threats in pristne ecosystems in Sri Lanka Arunachal Pradesh, India – G.K.V.P.T. Silva, W.A.D. Mahaulpatha & Anslem de Silva, Pp. 15004–15014 – Khilendra Singh Kanwal, Umeshkumar Lalchand Tiwari, Lod Yama & Mahendra Singh Lodhi, Pp. 15083–15086 Diversity and distributon of orchids of Goa, Western Ghats, India – Jeewan Singh Jalal, Pp. 15015–15042 Carex phacota, Spreng. (Cyperaceae): a new record for the central Western Ghats of Karnataka, India Short Communicatons – E.S.K. Udupa, H.U. Abhijit & K.G. Bhat, Pp. 15087–15088

Efcacy of oxyclozanide and levamisole treatment on the gastrointestnal Book review parasites in captve Lions Panthera leo – Dhareppa Ganager, Gotakanapura Sanjeevamurthy Mamatha, Asoor Compendium of Traded Indian Medicinal Plants Muralidhara, Nagappa Lakkundi Jaya & Beechagondahalli Papanna Shivashankar, – Reviewed by A. Rajasekaran, Pp. 15089–15090 Pp. 15043–15046

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