PLATINUM The Journal of Threatened Taxa (JoTT) is dedicated to building evidence for conservaton globally by publishing peer-reviewed artcles OPEN ACCESS online every month at a reasonably rapid rate at www.threatenedtaxa.org. All artcles published in JoTT are registered under Creatve Commons Atributon 4.0 Internatonal License unless otherwise mentoned. JoTT allows unrestricted use, reproducton, and distributon of artcles in any medium by providing adequate credit to the author(s) and the source of publicaton. Journal of Threatened Taxa Building evidence for conservaton globally www.threatenedtaxa.org ISSN 0974-7907 (Online) | ISSN 0974-7893 (Print) Note Larvae of the blow fly Caiusa testacea (Diptera: Calliphoridae) as egg predators of Polypedates cruciger Blyth, 1852 (Amphibia: Anura: Rhacophoridae) W.G.D. Chathuranga, K. Kariyawasam, Anslem de Silva & W.A.Priyanka P. de Silva 26 December 2020 | Vol. 12 | No. 17 | Pages: 17374–17379 DOI: 10.11609/jot.5740.12.17.17374-17379 For Focus, Scope, Aims, Policies, and Guidelines visit htps://threatenedtaxa.org/index.php/JoTT/about/editorialPolicies#custom-0 For Artcle Submission Guidelines, visit htps://threatenedtaxa.org/index.php/JoTT/about/submissions#onlineSubmissions For Policies against Scientfc Misconduct, visit htps://threatenedtaxa.org/index.php/JoTT/about/editorialPolicies#custom-2 For reprints, contact <[email protected]> The opinions expressed by the authors do not refect the views of the Journal of Threatened Taxa, Wildlife Informaton Liaison Development Society, Zoo Outreach Organizaton, or any of the partners. The journal, the publisher, the host, and the part- Publisher & Host ners are not responsible for the accuracy of the politcal boundaries shown in the maps by the authors. Member Threatened Taxa Journal of Threatened Taxa | www.threatenedtaxa.org | 26 December 2020 | 12(17): 17374–17379 ISSN 0974-7907 (Online) | ISSN 0974-7893 (Print) PLATINUM OPEN ACCESS DOI: htps://doi.org/10.11609/jot.5740.12.17.17374-17379 #5740 | Received 28 January 2020 | Final received 10 November 2020 | Finally accepted 04 December 2020 N o t e Larvae of the blow fy Caiusa testacea (Diptera: Calliphoridae) as egg predators of Polypedates cruciger Blyth, 1852 (Amphibia: Anura: Rhacophoridae) W.G.D. Chathuranga 1 , K. Kariyawasam 2 , Anslem de Silva 3 & W.A.Priyanka P. de Silva 4 1,2,4 Department of Zoology, Faculty of Science, University of Peradeniya, Peradeniya, Sri Lanka. 1 Postgraduate Insttute of Science, University of Peradeniya, Peradeniya, Sri Lanka. 3 No 15/1, Dolosbage Road, Gampola, Sri Lanka. 1 [email protected], 2 [email protected], 3 [email protected], 4 [email protected] (corresponding author) Habitat destructon and alteraton have been of terrestrial Rhacophoridae eggs (Rognes 2015). These identfed as the most detrimental causes of amphibian fies are one of the major reasons for embryo mortality decline (Kiesecker 2003). The efects of climate change of some rhacophorid genera, including Chiromants, and amphibian diseases, however, are emerging topics, Feihyla, Polypedates, and Rhacophorus (Lin & Lue 2000). and have taken increased atenton in conservaton So far, seven known species of Caiusa (C. borneoensis approaches regarding the amphibian fauna (Hayes Rognes, 2015, C. coomani Séguy, 1948, C. indica Surcouf, et al. 2010; Li et al. 2013). Predatory pressure during 1920, C. karrakerae Rognes, 2015, C. kurahashii Rognes, diferent life stages of amphibians is another factor 2015, C. violacea Séguy, 1925, and C. pooae Rognes, that signifcantly afects populatons (Chivers et al. 2015) have been identfed as foam nest predators and 2001; Blaustein et al. 2012). Diverse invertebrate and predators of jelly-like egg masses of anurans (Lin & Lue vertebrate fauna prey on eggs and tadpoles of aquatc 2000; Rognes 2015; Banerjee et al. 2018). The emerging and terrestrial nestng anurans (De Silva 2001a,b; Lingnau larvae of these fy species consume eggs and developing & Di-Bernardo 2006). According to Downie (1990), embryos in egg masses. There are knowledge gaps in terrestrial foam nests of Rhacophoridae have evolved to our understanding of the fy-anuran interactons and protect eggs and embryos from aquatc predators. Some the wider impact of these fies on anuran populaton vertebrates (e.g., monkeys and snakes) and invertebrates dynamics. (e.g., beetles, ephydrid fies, phorid fies, spiders, ants, Sri Lanka is a tropical country with more than 120 and blow fies), however, have been identfed as egg species of anurans, nearly 104 of which are endemic to the predators of anuran foam nests (Vonesh 2000; Rödel et country (De Silva & Wijayathilaka 2019). Approximately al. 2002; Menin & Giareta 2003; Lingnau & Di-Bernardo 83 (69%) of the reported species belong to the family 2006; Banerjee et al. 2018). Blow fies of the genus Caiusa Rhacophoridae, including arboreal foam nestng (Diptera: Calliphoridae) are one of the major predators Polypedates and Taruga species (Meegaskumbura et al. Editor: Daniel Whitmore, State Museum of Natural History Stutgart, Rosenstein, Germany. Date of publicaton: 26 December 2020 (online & print) Citaton: Chathuranga, W.G.D., K. Kariyawasam, A.D. Silva & W.A.P.P. de Silva (2020). Larvae of the blow fy Caiusa testacea (Diptera: Calliphoridae) as egg predators of Polypedates cruciger Blyth, 1852 (Amphibia: Anura: Rhacophoridae). Journal of Threatened Taxa 12(17): 17374–17379. htps://doi.org/10.11609/ jot.5740.12.17.17374-17379 Copyright: © Chathuranga et al. 2020. Creatve Commons Atributon 4.0 Internatonal License. JoTT allows unrestricted use, reproducton, and distributon of this artcle in any medium by providing adequate credit to the author(s) and the source of publicaton. Funding: Natonal Research Council Sri Lanka (Grant No: NRC 16-059) and Amphibian Specialist Group/IUCN/ SSC Seed Grant. Competng interests: The authors declare no competng interests. Acknowledgements: Financial assistance from Natonal Research Council Sri Lanka (Grant No: NRC 16-059) to W.A.P.P. de Silva. Amphibian Specialist Group/IUCN/ SSC Seed Grant to A. de Silva for threats to amphibians of Sri Lanka study. 17374 J TT Dipteran egg predators of Polypedates cruciger Chathuranga et al. 2010). The majority (more than 75%) of anuran species from uninfected ones. Three severely infected spawns in the country are categorized as threatened, mainly were carefully removed from the atached substrates due to anthropogenic actvites (Manamendra-Arachchi and brought to the Insectary, Department of Zoology, & Meegaskumbura 2012; De Silva & Wijayathilaka University of Peradeniya for further investgatons. 2019). Current conservaton approaches are mainly At the laboratory, the foam nests were placed in aimed at minimizing habitat destructon and other dechlorinated water in a tray and transferred to fne- adverse human actvites. Only a few studies, however, mesh mosquito rearing cages (50 × 50 × 50 cm) for have reported the efect of amphibian diseases on the maintenance of the fy colonies (at 25°C temperature, populaton structure of anurans in Sri Lanka (De Silva 75% relatve humidity, and 12 D: 12 L photoperiodicity). 1999; Rajakaruna et al. 2007; Jayawardena et al. 2010; Emerged fies were euthanized at -20°C in a freezer and De Silva 2011), and hardly any studies have investgated pinned for identfcaton. Morphological identfcaton predatory pressure on diferent life stages of amphibians was done using the standard taxonomic key in Rognes in the country. Morgan-Davies (1958) reported Caiusa (2015). indica as predatory in foam nests of Polypedates To confrm the identty of the dipteran species, DNA cruciger Blyth, 1852 (Anura: Rhacophoridae) in Sri barcoding was also performed. DNA was extracted from Lanka. According to De Silva & De Silva (2000), a species some of the collected fies following Livak (1984). The of Calliphoridae fy acts as an egg predator of P. cruciger mitochondrial Cytochrome Oxidase I (COI) gene was frogs, however, these authors did not provide a species- amplifed using the previously described primers C1-J- level identfcaton for the fies. Therefore, there are 1718F (5’-GGA GGA TTT GGA AAT TGA TTA GTT CC-3’) some literature gaps in informaton about predatory and C1-N-2191R (5’-CCC GGT AAA ATT AAA ATA TAA fies and their pressure on the developmental stages ACT TC-3’) (Simon et al. 1994). PCR amplifcaton was of anurans in Sri Lanka. Thus, detailed investgatons done in a thermal cycler (Techne-Flexigene, England) including systematc and quantfed studies to assess following Nolan et al. (2007). Positve PCR products were the damage caused by the egg predators to anuran eggs sequenced using an automatc DNA sequencer (Applied are important in relaton to conservaton actons. In this Biosystems Series 3500, U.S.A.) in the Department of study, we identfed natural dipteran predators of foam Molecular Biology and Biotechnology, University of nests of P. cruciger, an endemic Rhacophoridae species Peradeniya. The sequence trace fles were manually in Sri Lanka. Further, we quantfed the egg mortality inspected using MEGA V7 (Kumar et al. 2016) and low- of P. cruciger due to the infestaton of the predatory quality sequences were excluded from the analysis. The dipteran fy. DNA sequences were annotated using the GenBank The study was conducted from May 2019 to August database (htps://www.ncbi.nlm.nih.gov/) and BLASTn 2019, at two localites [Gampola (7.1500N, 80.5550E) and tool. The newly generated sequences were deposited in Peradeniya (7.2590N, 80.5970E)] in the Kandy District of GenBank under