21

THE STOMACH CONTENTS OF SOME ANURAN TADPOLES FROM THAILAND

Benjawan Moonasa1, Prapaiporn Thongproh1, Ekachai Phetcharat2, Wassana Kingwongsa3, Pramote Ratree4, Prateep Duengkae5, Theerasak Somdee6, Yodchaiy Chuaynkern1 & Chantip Chuaynkern1,*

ABSTRACT

The stomach contents of 18 tadpole in Thailand were studied. The tadpoles in this study consisted of one species of the family Bufonidae (Duttaphrynus melanostictus); one species of Dicroglossidae (); four species of Megophryidae (Megophrys carinense, Megophrys sp., Leptobrachium smithi and Leptolalax sp.); six species of ( butleri, Microhyla berdmorei, Microhyla heymonsi, Micryletta inornata, Kaloula pulchra and molossus); three species of Ranidae (Clinotarsus penelope, Hylarana erythraea and Sylvirana nigrovittata); and three species of Rhacophoridae (Polypedates leucomystax, Rhacophorus rhodopus and Rhacophorus jarujini). Tadpole specimens were collected by net from the following five areas: Phu Khiao Wildlife Sanctuary in Chaiyaphum Province, Yoddom Wildlife Sanctuary in Ubon Ratchathani Province, Phu Kradueng National Park in Loei Province, Umphang Wildlife Sanctuary in Tak Province, and Nam Nao National Park in Phetchabun Province. Specimens were dissected in order to remove their stomachs. All stomach contents were kept in order to create semi-permanent slides, which were observed under the light microscope. The results of tadpole stomach contents showed eight groups of food items, which included microalgae (25 genera), protozoa (3 genera), fungi like-organisms, rotifers (3 genera, including amictic eggs and bdelloid rotifers), diatoms (16 genera), arthropods, nematodes, and parts.

Keywords: , diet, food items, stomach analysis, Thailand

INTRODUCTION

Food is an important welfare factor for all living organisms. In , caecilians generally feed on earthworms and other invertebrates, while anurans and salamanders feed almost exclusively on insects (Vitt & Caldwell, 2009). Naturally, there is large diversity of food items available in their habitats, but the amphibians do not eat all of these prey items. Depending upon amphibian species and their foraging habits, only subsets of these available foods are eaten. Environment factors, such as physical and biological factors (or abiotic and biotic factors), influence the diversity of amphibian diets.

1 Department of Biology, Faculty of Science, Khon Kaen University, Mueang Khon Kaen, Khon Kaen, 40002, Thailand 2 Na Yung-Nam Som National Park, Udon Thani, 41380, Thailand 3 Center for Study Natural and Wildlife, Nam Yuen, Ubon Ratchathani, 34260, Thailand 4 Protected Area Regional Office 14 (Ubon Ratchathani), Department of National Parks, Wildlife and Plant Conservation, Mueang Ubon Ratchathani, Ubon Ratchathani, 63000, Thailand 5 Department of Forest Biology, Faculty of Forestry, Kasetsart University, Chatuchak, Bangkok, 10900, Thailand 6 Department of Microbiology, Faculty of Science, Khon Kaen University, Mueang Khon Kaen, Khon Kaen, 40002, Thailand * Corresponding author: [email protected]

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 22

To determine what amphibians eat, researchers try to use various methods and analyses to obtain their results (e.g., Solé et al., 2005; Chuaynkern et al., 2009; Camera et al., 2014; Ponpituk et al., 2015; Goldberg et al., 2018). Based on the items that we presently know that amphibians eat, we are still far from fully understanding their complete diets because what is known is mainly based on the diets of adult (or juvenile) amphibians. In amphibian tadpoles, food is very important for development (Kupferberg, 1997; Alford, 1999), which influences the size and shape of the tadpoles. However, little information about their food items has been published. Unlike the adult amphibians, the tadpoles generally feed on fungi, viruses, bacteria, algae, protists, helminths, plant cells, and on other tadpoles (Kupferberg et al., 1994; Hoff et al., 1999; Goldberg et al., 2018). Moreover, cannibalism in anuran larvae are also reported in several species (see e.g., Heusser, 1970; Jungfer & Schiesari, 1995). Approximately 178 amphibian species are known in Thailand, which is 10% of the known global species (Frost, 2018; Phusaensri et al., 2018). Among them, only some anuran species have had their food items reported on in detail (Chuaynkern et al., 2009; Ponpituk et al., 2015). However, for the tadpoles of the amphibians, almost nothing is known about their food items. Therefore, this present work has reported on the prey items of 18 anuran tadpoles based upon specimens collected from five localities of Thailand.

MATERIALS AND METHODS

From the following five localities in Thailand (Figure 1), the tadpole specimens were collected from nature by using dip-nets: Phu Kradueng National Park (Loei Province), Yoddom Wildlife Sanctuary (Ubon Ratchathani Province), Phu Khiao Wildlife Sanctuary (Chaiyaphum Province), Umphang Wildlife Sanctuary (Tak Province), and Nam Nao National Park (Phetchabun Province). The tadpoles were euthanized using 70% ethyl alcohol and were later preserved in a mixture solution of 10% formaldehyde and 70% ethyl alcohol at a ratio of 1:1 (followed Inthara et al., 2009). These tadpoles (Table 1) were catalogued and deposited in the Khon Kaen University Vertebrates Collection (KKUC) in Khon Kaen Province in northeastern Thailand. The identification of tadpoles was made by comparing the samples to relevant taxonomic works (Inthara, 2000; Grosjean et al., 2003, 2015; Taksintum, 2003; Meewattana, 2005; Stuart et al., 2006; Danisawat et al., 2010; Aran et al., 2012; Kaewtongkum et al., 2014a,b; Chuaynkern et al., 2017; Thongproh et al., 2018). The developmental stages of the tadpole specimens were determined according to the work of Gosner (1960). In order to investigate the stomach contents, tadpoles were selected, which were between Gosner’s stages of 31 to 39. Preparation of semi-permanent slides was modified from the protocol proposed by Somnark et al. (2011). The gut was excised by opening the tadpoles’ bellies using a surgical scissors. Later, the gut was opened, and all of the stomach contents were removed and placed into a Petri dish to which 70 % ethyl alcohol was added to preserve them. Then using a dropper was used to collect the solution of stomach contents, one drop was placed on the slide along with one drop of glycerin. Finally, the slide was covered with a cover slip and was sealed with nail polish at the edges of the cover slip. The stomach contents found on the semi-permanent slide were identified under a compound microscope (Zeiss) at 10x and 40x. Photographs were taken of all

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 23 the food items using a compound microscope (Zeiss). In order to identify these prey items, the photographs were compared with the standard literature review, and for some of the unidentified items, specialists were consulted.

Figure 1 Map showing collected sites of larval specimens. 1=Umphang Wildlife Sanctuary (Tak Province), 2=Phu Kradueng National Park (Loei Province); 3=Nam Nao National Park (Phechabun Province); 4=Phu Khiao Wildlife Sanctuary (Chaiyaphum Province); 5=Yoddom Wildlife Sanctuary (Ubon Ratchathani Province).

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 24

Table 1 A list of tadpole specimens used in the present work. Map of localities was presented in Figure 1. Scientific names followed Frost (2018). Abbreviation: KD=Phu Kradueng NP, KKU=Khon Kaen Unviversity, NP=National Park, PK=Phu Khiao WS, PU=Protected Unit, WS=Wildlife Sanctuary, YC=Yodchaiy Chuaynkern, YD=Yoddom WS.

Family/Species Number Stage No. of Locality Habitat specimens Family Bufonidae Duttaphrynus melanosctitus PK 007 31 3 Phu Khiao WS., “Temporary pond”, Chaiyaphum, Thung Kra Mung northeastern Thailand Family Dicroglossidae Fejervarya limnocharis PK 016 (R2) 31 3 Phu Khiao WS., “Temporary pond”, Chaiyaphum, Near Mor Chang Tai northeastern Thailand Family Megophryidae Megophrys carinense YC 122 36 1 Umphang WS., “Permanent forest steram”, Tak, western Thailand Huay Umphang Kee, Pa Do Tha PU. Megophrys sp. YC 157 36 1 Umphang WS., “Permanent forest stream”, Tak, western Thailand Tee Lor Zoo PU Leptobrachium smithi PK 009 (R2) 31,35,36 3 Phu Khiao WS., “Permanent forest stream”, Chaiyaphum, Huay Pong Hi northeastern Thailand Leptobrachium smithi YC 00814 37 1 Umphang WS., “Permanent forest stream”, Tak, western Thailand Huay Sae Dor Soup Gro Leptolalax sp. YC 00819 36 1 Umphang WS., “Permanent forest stream”, Tak, western Thailand Huay Tee Bue La Gro Family Microhylidae Glyphoglossus molossus PK 025 (R2) 34,35 3 Phu Khiao WS., “Temporary pond”, Chaiyaphum, near the Palace northeastern Thailand Kaloula pulchra KD 052 31,32,33 3 Phu Kradueng NP., “Temporary pond”, Loei, northeastern Lan Phra Kaew Thailand Microhyla berdmorei YC 00721 37 3 Umphang WS., “Permanent forest stream”, Tak, western Thailand Huay Tee Pha Do Microhyla berdmorei PK 006 33,34,35 3 Phu Khiao WS., “Temporary forest stream”, Chaiyaphum, Huay Kam Yan northeastern Thailand Microhyla butleri PK 020 (R2) 34,36,37 3 Phu Khiao WS., “Temporary pond”, Chaiyaphum, Thailand near Mor Chang Tai Microhyla butleri YD 0275 38 3 Yoddom WS., “Temporary pond”, Ubon Rachathani, Huay Chan Daeng PU northeastern Thailand Microhyla heymonsi PK 024 (R2) 31,32,33 3 Phu Khiao WS., “Temporary pond”, Chaiyaphum, Nong Paak northeastern Thailand Micryletta inornata PK 006 (R2) 35,36,37 3 Phu Khiao WS., “Temporary pond”, Chaiyaphum, Nong Paak northeastern Thailand Family Ranidae Clinotarsus penelope YC 00816 39 1 Umphang WS., Tak, “Permanent forest stream”, western Thailand Huay Sae Dor Soup Gro

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 25

Table 1 (Continued).

Family/Species Number Stage No. of Locality Habitat specimens Hylarana erythraea PK 024 35,36 3 Phu Khiao WS., “Temporary pond”, Chaiyaphum, between forest road to northeastern Thailand Bueng Mon Sylvirana nigrovittata PK 023 (R2) 31,36 3 Phu Khiao WS., “Artificial pond”, Chaiyaphum, Head quarter. northeastern Thailand Family Rhacophoridae Polypedates leucomystax PK 019 (R2) 32,35 3 Phu Khiao WS., “Temporary pond”, Chaiyaphum, near Mor Chang Tai northeastern Thailand Rhacophorus jarujini KKUC 01937 36 1 Yoddom WS., “Temporary pond” in forest Ubon Rachathani, northeastern Thailand Rhacophorus rhodopus KKU 02371 36 1 Nam Nao NP, “Temporary pond”, Head Phetchabun, quarter northeastern Thailand

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 26

Figure 2 Larval photographs in this study (A= Duttaphrynus melanostictus, B= Fejervarya limnocharis, C= Megophrys carinense, D= Leptobrachium smithi, E= Megophrys sp., F = Leptolalax sp.).

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 27

Figure 3 Larval photographs in this study (A= Kaloula pulchra, B= Microhyla butleri, C= Microhyla berdmorei, D= Microhyla heymonsi, E= Glyphoglossus molossus, F= Micryletta inornata).

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 28

Figure 4 Larval photographs in this study (A= Clinotarsus penelope, B= Hylarana erythraea, C= Sylvirana nigrovittata, D= Polypedates leucomystax, E= Rhacophorus rhodopus, F= Rhacophorus jarujini).

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 29

RESULTS

The results of the stomach contents from the 18 tadpole species from the 6 families showed eight groups of food items, including Microalgae (25 genera), Protozoa (3 genera), Fungi like-organisms, Rotifers (3 genera, including amictic egg and Bdelloid rotifers), diatoms (16 genera), arthropods, nematodes, and plant parts (see Table 2; Figures 5-9.) Family Bufonidae: Duttaphrynus melanostictus found microalgae, Protozoa, fungi like- organisms, rotifers, diatoms, arthropods and plant materials. Family Dicroglossidae: Fejervarya limnocharis found microalgae, fungi like-organisms, rotifers, diatoms, arthropods, and plant fragments. Family Megophryidae: 1) Megophrys carinense found microalgae, fungi like-organisms, diatoms, Protozoa, arthropods, and nematodes; 2) Leptobrachium smithi (from Phu Khiao Wildlife Sanctuary) found microalgae, fungi like-organisms, Protozoa, diatoms, rotifers, arthropods, and plant fragments; 3) Leptobrachium smithi (from Umphang Wildlife Sanctuary) found microalgae, fungi like-organisms, diatoms, Protozoa, arthropods, nematodes, and leaf fragments; 4) Megophrys sp. found fungi like- organisms and arthropods; 5) Leptolalax sp. found fungi like-organisms, diatoms, Protozoa, and arthropods. Family Microhylidae: 1) Glyphoglossus molossus found microalgae, fungi like-organisms, Protozoa and plant fragments; 2) Kaloula pulchra found microalgae, Cyanobacteria, Protozoa and plant fragments; 3) Microhyla berdmorei (from Phu Khiao Wildlife Sanctuary) found microalgae, fungi like-organisms, rotifers, copepod fragments, Protozoa, diatoms, arthropods, and plant fragments; 4) Microhyla berdmorei (from Umphang Wildlife Sanctuary) found diatoms and arthropods; 5) Microhyla butleri (from Phu Khiao Wildlife Sanctuary) found microalgae, fungi like-organisms, Protozoa, diatoms and plant fragments; 6) Microhyla butleri (from Yoddom Wildlife Sanctuary) found microalgae, fungi like-organisms, diatoms, and plant fragments; 7) Microhyla heymonsi found microalgae, fungi like- organisms, arthropods, and plant fragments; 8) Micryletta inornata found microalgae, Protozoa, fungi like-organisms, and plant fragments. Family Ranidae: 1) Clinotarsus penelope found microalgae, diatoms, protozoa, and arthropods; 2) Hylarana erythraea found microalgae, fungi like-organisms, protozoa, rotifers, diatoms, and plant fragments; 3) Sylvirana nigrovittata found microalgae, protozoa, fungi like-organisms, and plant fragments. Family Rhacophoridae: 1) Polypedates leucomystax found microalgae, fungi like-organisms, rotifers, copepods, diatoms, and plant fragments; 2) Rhacophorus rhodopus found microalgae, fungi like-organisms, arthropods, protozoa, and nematodes; 3) Rhacophorus jarujini found microalgae, fungi like-organisms, diatoms, protozoa, and arthropods.

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 30

วารสารสัตว์ป่าเมืองไทย25 ปีที่ Table 2 Stomach content items of 18 anuran tadpoles from Thailand.

Phylum Class Order Family Genus mel lim car smi lax meg mol pul ber but hey ino pen ery nig leu jar rho Plantae Conjugatophyceae Closteriaceae Closterium + + + + + + + + + + + Charophyta Conjugatophyceae Desmidiales Cosmarium + + + + Charophyta Conjugatophyceae Desmidiales Desmidiaceae Euastrum + +

Charophyta Conjugatophyceae Desmidiales Desmidiaceae +

พ.ศ.2561 Charophyta Conjugatophyceae Desmidiales Desmidiaceae Pleurotaenium + Charophyta Conjugatophyceae Desmidiales Desmidiaceae Spondylosium + Charophyta Conjugatophyceae Desmidiales Desmidiaceae Staurastrum + + Charophyta Conjugatophyceae Desmidiales Desmidiaceae Triploceras +

Charophyta Conjugatophyceae Desmidiales Desmidiaceae Xanthidium +

Charophyta Conjugatophyceae Zygnematales Mesotaeniaceae Netrium + Charophyta Conjugatophyceae Zygnematales Zygnemataceae Mougeotia + Charophyta Conjugatophyceae Zygnematales Zygnemataceae Spirogyra + +

Charophyta Trebouxiophyceae Chlorellales Chlorellaceae Geminella + + + Chlorophyta Chlorophyceae Oedogoniales Oedogoniaceae Oedogonium + Chlorophyta Ulvophyceae Ulotrichales Ulotrichaceae Ulothrix + + + + + Chlorophyta Ulvophyceae Cladophorales Cladophoraceae Cladophora + Chlorophyta Chlorophyceae Sphaeropleales Selenastraceae Monoraphidium + - - - - Plant part + + + + + + + + + + + +

Bacteria Cyanobacteria Cyanophyceae Synechococcales Pseudanabaenaceae Bacteria + Cyanobacteria Cyanophyceae Synechococcales Pseudanabaenaceae Pseudanabaena +

Cyanobacteria Cyanophyceae Oscillatoriales Oscillatoriaceae Oscillatoria + + + + + + + +

Journal of WildlifeThailandin Journal25 of Vol. Cyanobacteria Cyanophyceae Oscillatoriales Oscillatoriaceae Phormidium + Cyanobacteria Cyanophyceae Nostocales Nostocaceae Anabaena + Cyanobacteria Cyanophyceae Spirulinales Spirulinaceae Spirulina + + + + Protozoa Euglenozoa Euglenophyceae Euglenales Euglenaceae Trachelomonas + + + + + + + + + + + Euglenozoa Euglenophyceae Euglenales Phacaceae Phacus + + + + + + + Euglenozoa Euglenophyceae Euglenales Euglenaceae Euglena + + + + + + + Amoebozoa Tubulinea Arcellinida Arcellidae Arcella + + + + + + + Amoebozoa Tubulinea Arcellinida Centropyxidae Centropyxis + Paramecium Ciliophora Oligohymenophorea Peniculida Parameciidae Paramecium + + + Fungi

, 2018 , Fungi like-organisms 1 - - - - + + + + + + + +

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 31

วารสารสัตว์ป่าเมืองไทย25 ปีที่ Table 2 (Continued).

Phylum Class Order Family Genus mel lim car smi lax meg mol pul ber but hey ino pen ery nig leu jar rho FLOgi like-organisms 2 - - - - + + + + + + + + + + + + + Fungi like-organisms 3 - - - - + + Rotifer Rotifera Eurotatoria Ploima Brachionidae Plationus patulus +

Rotifera Eurotatoria Ploima Lepadellidae Lepadella +

พ.ศ.2561 Rotifera Eurotatoria Ploima Lecanidae Lecane hamata + + Rotifera Eurotatoria - - Bdelloid rotifers + + + Rotifera - - - Amictic egg +

Diatom Bacillariophyta Bacillariophyceae Naviculales Pinnulariaceae Pinnularia + + + + + + Bacillariophyta Bacillariophyceae Naviculales Naviculaceae Navicula + + + Bacillariophyta Bacillariophyceae Naviculales Sellaphoraceae Sellaphora + Bacillariophyta Bacillariophyceae Naviculales Neidiaceae Neidium + Bacillariophyta Bacillariophyceae Naviculales Stauroneidaceae Stauroneis + + Bacillariophyta Bacillariophyceae Naviculales Stauroneidaceae Craticula + Bacillariophyta Bacillariophyceae Surirellales Surirellaceae Surirella + + + +

Bacillariophyta Bacillariophyceae Eunotiales Eunotiaceae Eunotia + Bacillariophyta Bacillariophyceae Cocconeidales Cocconeidaceae Cocconeis + Bacillariophyta Bacillariophyceae Mastogloiales Achnanthaceae Achnanthes +

Bacillariophyta Bacillariophyceae Fragilariales Fragilariaceae Synedra + + + + Bacillariophyta Bacillariophyceae Tabellariales Tabellariaceae Diatoma + Bacillariophyta Bacillariophyceae Fragilariales Fragilariaceae Fragilaria + +

Bacillariophyta Bacillariophyceae Cymbellales Cymbellaceae Cymbella + + +

Journal of WildlifeThailandin Journal25 of Vol. Bacillariophyta Bacillariophyceae Naviculales Naviculaceae Caloneis + Bacillariophyta Bacillariophyceae Cymbellales Gomphonemataceae Geissleria + + Arthopoda Arthropoda SP. Crustacea SC. Copepoda Copepod material + + Arthropoda Insecta Lepidoptera - Butterfly wing scales + + + + + + + + Arthropoda Arachnida - - Arachnid material + Arthropoda Insecta - - Insect material + Arthropoda Entognatha SC. Collembola - Collembora material + Arthropoda Insecta Odonata - Odonata material + Arthropoda Insecta - - Claw + Arthropoda Insecta - - Arthropod material + Arthropoda Insecta Diptera Chironomidae Chironomidae larva +

, 2018 , Nematoda - - - Nematode + + +

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 32

Note: SP=Subphylum, SC=Subclass, mel=Duttaphrynus melanostictus, lim=Fejervarya limnocharis, car=Megophrys carinense, smi=Leptobrachium smithi, lax=Leptolalax sp., meg= Megophrys sp., mol=Glyphoglossus molossus, pul=Kaloula pulchra, ber=Microhyla berdmorei, but=Microhyla butleri, hey=Microhyla heymonsi, ino=Micryletta inornata, pen=Clinotarsus penelope, ery=Hylarana erythraea, nig=Sylvirana nigrovittata, leu=Polypedates leucomystax, jar=Rhacophorus jarujini, rho=Rhacophorus rhodopus.

DISCUSSION

The results from examining the stomach contents of some tadpoles showed that the tadpoles were herbivorous and omnivorous. In fact, microalgae was the most abundant item in all of the tadpoles’ stomachs, followed by diatoms and fungi like-organisms, respectively. These findings were similar to the food items mentioned in a study by Heyer (1973), who found that the food items of eight anuran tadpoles (Glyphoglossus molossus, Microhyla fissipes, Microhyla butleri, Microhyla heymonsi, Kaloula pulchra, Duttaphrynus melanostictus, Fejervarya limnocharis and Sylvirana nigrovittata) at the Sakaerat Environmental Research Station in Nakhon Ratchasima, Thailand, which was filled with algae and diatoms. In addition, Singh et al. (2014) reported that the food items of Leptobrachium smithi tadpole (from Rosekandy Tea Estate, Cachar, Assam) consisted of diatoms, algae, and protozoa. Regarding our results, the tadpoles were separated into two groups: 1) a keratinized group and 2) a non-keratinized group. Firstly, the keratinized group included ten species (Fejevarya limnocharis, Sylvirana nigrovittata, Hylarana erythraea, Duttaphrynus melanostictus, Leptobrachium smithi, Polypedates leucomystax, Clinotarsus penelope, Leptolalax sp., Rhacophorus rhodopus and Rhacophorus jarujini) which fed on algae, protozoa, fungi like-organisms, rotifers, diatoms, copepods, nematodes, arthropods (butterfly wing scales, insect fragments Collembora fragments, Chironomidae larvae, and the claws and bodies of arthropods), and plant parts. In this work, it was found that Polypedates leucomystax had fed on algae. This finding is in agreement with Inthara (2000), who suggested that Polypedates leucomystax present as keratinized and has a beak which causes their feeding behaviors to be predator-like, allowing them to feed on algae growing on substrates. Conversely, the non-keratinized tadpoles consisted of eight species (Kaloula pulchra, Microhyla butleri, Microhyla berdmorei, Microhyla heymonsi, Glyphoglossus molossus, Micryletta inornata, Megophrys carinense and Megophrys sp.), which had fed on algae, protozoa, fungi-like organisms, rotifers, diatoms, copepods, arthropods (butterfly wing scales, arachnid fragments, and wing fragments of Odonata), nematodes, and on plant parts. In addition, the habitat itself is a factor that contributes to differences in food items. Khan (1999) suggested that tadpoles usually feed on the food that is in their habitat and that ecological factors can also influence the food selection of tadpoles in their natural habitats (Duellman & Trueb, 1994). In case of insufficient food in particular habitat, a cannibalism could be occurred (Alford, 1999). At Yoddom Wildlife Sanctuary (Ubon Ratchathani Province), the Rhacophorus jarujini were observed to eat other dead Rhacophorus jarujini tadpoles (Y. Chuaynkern: personal observation). As our knowledge about food items in anuran tadpoles are little known, this paper provides the preliminary investigation on anuran diets. However, further study in various amphibian tadpoles is essential to full-fill our gap of knowledge about dietary of this vertebrate group.

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 33

Figure 5 Larval Food items found in 18 tadpoles. A=Closterium sp., B=Cosmarium sp., C= Euastrum sp., D=Micrasterias sp., E=Pleurotaenium sp., F=Spondylosium sp., G=Staurastrum sp., H=Triploceras sp., I= Xanthidium sp., J. Netrium sp., K=Mougeotia sp., L=Spirogyra sp., M=Geminella sp., N=Algae, O=Oedogonium sp.

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 34

Figure 6 Food items found in 18 tadpoles. A=Ulothrix sp., B=Cladophora sp., C= Monoraphidium sp., D=Oscillatoria sp., E=Phormidium sp., F=Anabaena sp., G= Spirulina sp., H=Pseudanabaena sp., I= Trachelomonas sp., J=Phacus sp., K=Euglena sp., L=Arcella sp., M= Centropyxis sp., N=Paramecium sp., O=Fungi like-organisms.

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 35

Figure 7 Food items found in 18 tadpoles. A= Fungi like-organisms, B=Plationus patulus, C=Lepadella sp., D= Lecane hamata, E=Bdelloid rotifers, F=Amictic egg, G= Pinnularia sp., H=Navicula sp., I=Sellaphora sp., J=Neidium sp., K=Stauroneis sp., L= Craticula sp., M=Surirella sp., N=Eunotia sp., O= Cocconeis sp.

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 36

Figure 8 Food items found in 18 tadpoles. A= Achnanthes sp., B=Synedra sp., C= Diatoma sp., D=Fragilaria sp., E= Cymbella sp., F=Caloneis sp., G=Geissleria sp., H=Copepod material, I=Butterfly wing scales, J=Arachnid material, K=Insect material, L=Collembora material, M=Odonata material, N=Claw, O=Chironomidae larva.

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 37

Figure 9 Food items found in 18 tadpoles. A=nematode, B,C= plant part.

ACKNOWLEDGEMENTS

We would like to thank the following institutions for facilitating this work: 1) Khon Kaen University (Department of Biology and Department of Microbiology, Faculty of Science), 2) Department of National Park, Wildlife and Plant Conservation, 3) Kasetsart University (Department of Forest Biology, Faculty of Forestry). In permission, we would like to thank the Department National Parks, Wildlife and Plant Conservation. For granting, the authors would like to express our grateful to the Faculty of Science (KKU) and the Research and Technology Transfer Affairs (KKU). For field works, we thank assistance from staffs of Yoddom WS, Phu Khiao WS, Umphang WS, Nam Nao NP and Phu Kradueng NP during field works. For assistance during the laboratory works, we thank Nattaporn Plangklang, Kamonwan Koompoot, Rachaneekorn Ramlee, Benjamart Suksai and Sirikamon Phlaingam. Our special thanks go to Asst. Prof. Sujeephon Athibai and Yuwadee Ponpituk for identification assistant. Finally, we thank the members of KKU Herpetological Laboratory for all supporting this work in various ways.

REFERENCES

Alford, R.A. 1999. Ecology: Resource use, competition, and predation. Pp. 240–278. In Tadpoles: The Biology of Anuran Larvae. R.W. McDiarmid & R. Altig (eds.). The University of Chicago Press, Chicago and London. Aran, S., C. Chuaynkern, S. Duengjai & Y. Chuaynkern. 2012. Morphology of some tadpoles in Khon Kaen University, Khon Kaen Province. Journal of Wildlife in Thailand 19(1): 41–73. Camera, B.F., D. Krinski & I.A. Calvo. 2014. Diet of the Neotropical Leptodactylus mystaceus (Anura: Leptodactylidae). Herpetology Notes 7: 31–36. Chuaynkern, Y., A. Wongwai, P. Duengkae & S. Hasin. 2009. Kalophrynus interlineatus: Diet. Herpetological Review 40(2): 205. Chuaynkern, C., D. Machan, P. Duengkae & Y. Chuaynkern. 2017. External and buccal morphologies of Glyphoglossus guttulatus, Microhyla butleri and Microhyla pulchra (Anura: Microhylidae). Journal of Wildlife in Thailand 24(1): 1–12.

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 38

Danaisawat, P., A. Pradatsundarasan & W. Khonsue. 2010. Morphological character of some tadpole from Khao Sip Ha Chan proposed national park, Chanthaburi Province. Journal of Wildlife in Thailand 17(1): 64–103. Duellman, W.E. & L. Trueb. 1994. Biology of Amphibians. The John Hopkins University Press, Baltimore, USA. Frost, D.R. 2018. Amphibian Species of the World: an Online Reference. Version 6.0. Available Source: http://research.amnh.org/herpetology/amphibia/index.html, November 3, 2018. Goldberg, S.R., C.R. Bursey & F. Kraus. 2018. Helminths of the green and black poison frog, Dendrobates auratus (Anura, Dendrobatidae) from Hawaii. Alytes 35(1–4): 39–42. Gosner, K.L. 1960. A simplified table for staging anuran embryos and larvae with notes on identification. Herpetologica 16(3): 183–190. Grosjean, S., M. Perez & A. Ohler. 2003. Morphology and buccopharyngeal anatomy of the tadpole of Rana (Nasirana) alticola (Anura: Ranidae). Raffles Bulletin of Zoology 51: 101–107. Grosjean, S., S. Bordoloi, Y. Chuaynkern, P. Chakravarty & A. Ohler. 2015. When young are more conspicuous than adults: a new ranid species (Anura: Ranidae) revealed by its tadpole. Zootaxa 4058(4): 471–498. Heusser, H. 1970. Laich-Fressen durch Kaulquappen als mögliche Ursache spezifischer Biotoppräferenzen und kurzer Laich-zeiten bei europäischen Froschlurchen (Amphibia, Anura). Oecologia 4: 83–88. Heyer, W. 1973. Ecological interactions of frog larvae at a seasonal tropical location in Thailand. Journal of Herpetology 7(4): 337–361. Hoff, K.S., R.A. Blaustein, R.W. McDiarmid & R. Altig. 1999. Behavior: Interactions and their consequences. Pp. 215–239. In Tadpoles: The Biology of Anuran Larvae. W.R. McDiarmid & R. Altig (eds.). The University of Chicago Press, Ltd., London. Inthara, C. 2000. Study on mouthpart structures in relation to feeding behavior of some tadpole species. MS Thesis, Kasetsart University, Bangkok [in Thai with English Abstract]. Inthara, C., Y. Chuaynkern, P. Duengkae & S. Grosjean. 2009. The tadpole of Quasipaa fasciculispina (Inger, 1970) from southeastern Thailand, with the description of its buccal anatomy. Alytes 26(1–4): 86–96. Jungfer, K.-H., & L.C. Schiesari. 1995. Description of a central Amazonian and Guianan tree frog, genus Osteocephalus (Anura, Hylidae), with oophagous tadpoles. Alytes 13: 1–13. Kaewtongkum, N., C. Chuaynkern, P. Thongproh, Y. Chuaynkern, E. Phetcharat, W. Maiprom, P. Ratree & P. Duengkae. 2014a. Buccal description of Rhacophorus jarujini Matsui and Panha, 2006 from Northeastern Thailand. In Proceedings of the 40th Congress on Science and Technology of Thailand (STT40). December 2–4, 2014, Hotel Pullman, Khon Kaen, Thailand. Kaewtongkum, N., Y. Chuaynkern, S. Duangjai, P. Ratree, M. Kamsook, S. Makchai, S. Sangarang, P. Duengkae & C. Chuaykern. 2014b. Morphological and buccal anatomies of Megophryidae tadpoles from Umphang District, Tak Province. In Proceedings of the 4th TST Conference. May 23-25, 2014, Faculty of Science, Naresuan University, Phitsanulok, Thailand. Khan, M.S. 1999. Food particle retrieval in amphibian tadpoles. Zoos’ Print Journal 14(5): 17–20.

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018 39

Kupferberg, S.J. 1997. The role of larval diet in anuran metamorphosis. American Zoologist 37: 146–159. Kupferberg, S.J., J.C. Marks & M.E. Power. 1994. Effects of variation in natural algal and detrital diets on larval anuran (Hylar egilla) life history traits. Copeia 2: 446–457. Meewattana, P. 2005. Morphological characters for identification of anuran tadpoles in Khao Luang National Park, Khlong Nakha and Hala-Bala Wildlife Sanctuary. PhD Dissertation, Kasetsart University, Bangkok. Phusaensri, S., T. Kaewboribut, S. Phummisutthigoon, N. Kaewtongkum, M. Youjaruen, P. Tongpun, H. Khongcharoensuk, P. Nurngsomsri, C. Chuaynkern, P. Duengkae & Y. Chuaynkern. 2018. Theloderma petilum (Anura: Rhacophoridae): a new country record for Thailand. Alytes 36(1–4): 289–299. Ponpituk, Y., R. Songchan, P. Duengkae, Y. Chuaynkern & S. Hasin. 2015. Diet of amphibians in Hill Evergreen Forest at Doi Suthep-Pui National Park, Chiang Mai Province and Phu Luang Wildlife Sanctuary, Loei Province. In Proceedings of the Thai Forest Ecological Research Network, T- FERN. January 22-23, 2015, Naresuan University, Phitsanulok, Thailand. Singh, P., M. Dey, & S.N. Ramnujam. 2014. Assessing feeding habits of tadpoles of tadpoles of Leptobrachium smithi Matsui, Nabhitabhata and Panha, 1999 during different development stage: A qualitative and quantitative study from Rosekandy tea estate, Cachar, Assam. International Journal of Scientific and Research Publications 4(5): 1–6. Solé, M., O. Beckmann, B. Pelz, A. Kwet & W. Engels. 2005. Stomach-flushing for diet analysis in anurans: an improved protocol evaluated in a case study in Araucaria forests, southern Brazil. Studies on Neotropical Fauna and Environment 40(1): 23–28. Somnark, R., N. Sangpradub & C. Hanjavanit. 2011. Stomach content of the Grey feather back (Notopterus notopterus (Pallas, 1780)) and the Bagrid catfish (Mystus mysticetus Roberts, 1992) in Kaeng Lawa, Khon Kaen Province, northeastern Thailand. Laos Journal on Applied Science 2(1): 491–498. Stuart, B.L., Y. Chuaynkern, T. Chan-ard & R.F. Inger. 2006. Three new species of and a new tadpole from eastern Thailand. Fieldiana, Zoology New Series 111: 1–19. Taksintum, W. 2003. Diversity of mature frogs and their tadpoles in Khlong Sang Wildlife Sanctuary. MS Thesis, Kasetsart University, Bangkok [in Thai with an English Abstract]. Thongproh, P., M. Youjaroen, Y. Chuaynkern, C. Chuaynkern, P. Ratree, P. Duengkae, E. Phetcharat, W. Maiprom, W. Ungprombundith & T. Wongpaiseart. 2018. On the identities of Rhacophorus jarujini Matsui and Panha, 2006 and Rhacophorus orlovi Ziegler and Köhler, 2001 (Amphibia, Anura, Rhacophoridae) from Thailand. Maejo International Journal of Science and Technology 12(01): 36–50. Vitt, J.V. & J.P. Caldwell. 2009. Herpetology: An Introductory Biology of Amphibians and Reptiles. Third Edition. Academic Press, Burlington, Massachusetts.

วารสารสัตว์ป่าเมืองไทย ปีที่ 25 พ.ศ. 2561 Journal of Wildlife in Thailand Vol. 25, 2018