Abstract

Dear participants of the First Asian Simuliidae Symposium, On behalf of the Asian Simuliidae Symposium committee and MahasarakhamUniversity, I would like to welcome you to northeastern Thailand, which is the location ofMahasarakham University. It is our pleasure and honor to be the host for the First AsianSimuliidae Symposium. Asian Simuliidologists have increasingly become a center forSimuliidae study, as has been recognized by the number of high standard publications inseveral academic journals. This symposium will bring together colleagues to share ideasand knowledge about research into Simuliidae in Asia including expanded networks,partnerships and collaborations between researchers. Although the name of the symposiumis Asian Simuliidae, it is not restricted to only Asian Simuliidae workers. We are verywelcoming to all participants from around the world. We thank the Faculty of Science and Mahasarakham University for financial and othervaluable support for the organization of this symposium. We hope that the first AsianSimuliidae symposium will be marked as the starting point for future collaboration onSimuliidae.Organizing Committee

2 Program25th July 2016 Registration08.30 – 09.00 Opening of the Asian Simuliidae Symposium09.00 – 09.3009.30 – 10.15 “What a wonderful fauna of black flies (Diptera: Simuliidae) in the Oriental Region”10.15 – 10.30 Hiroyuki Takaoka, University of Malaya, Malaysia10.30 – 11.00 Coffee Break11.00 – 11.30 “Assemblage structure of larval black fly (Simuliidae: Diptera)11.30 – 13.00 species in South India”13.00 – 13.30 Sankarappan Anbalagan, Alagappa University, India13.30 – 14.00 “Cryptic diversity in human-biting black flies (Diptera: Simuliidae) in Thailand”14.00 – 14.15 Pairot Pramual, Mahasarkham University, Thailand14.15 – 16.0018.00 – 20.00 Lunch “The Biodiversity of Black flies (Diptera: Simuliidae) in Indonesia” Upik Kesumawati Hadi, Bogor Agricultural University, Indonesia “Faunas of blackflies and administrative borders” SergeyV. Aibulatov, Zoological Institute of the Russian Academy of Sciences, Russia Coffee Break Poster Session Conference Dinner The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

26th July 2016 309.00 – 09.30 “Advances in Simuliidology and Changes in Geographic Centers09.30 – 10.00 of Simuliid Research” Peter H. Adler, Clemson University, USA10.00 – 10.15 “Symbionts of Larval black flies (Diptera: Simuliidae) in10.15 – 12.00 Thailand”12.00 – 13.00 Sanaea Jitklang, Mahidol University, Thailand13.00 – 14.30 Coffee Break Poster Session14.30 – 14.4514.45 – 16.30 Lunch18.00 – 20.00 Round table discussion for future research collaboration on Asian27th July 2016 Simuliidae07.00 Coffee Break Campus tour (visiting Medicinal Mushroom Museum / Center of Excellence for Silk Innovation / Palaeontological Research and Education Centre Conference Dinner Excursion: Elephant Village, Surin Province The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

4 ABSTRACTSOral Presentation The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

5 What a wonderful fauna of black flies (Diptera: Simuliidae) in the Oriental Region Hiroyuki Takaoka1,2,3 1Institute of Biological Sciences, Faculty of Science, University of Malaya, Malaysia; 2Professor Emeritus, Oita University, Japan; 3Distinguished fellow, The Japanese Society of Tropical Medicine, Japan ABSTRACTOwing to the recent increasing interest and activity in research of black flies, the fauna ofblack flies in the Oriental Region, which was poorly studied before 1970, and once rankedlast among the six zoogeographical regions in number of species, has dramatically emergedas the second largest in the number of the constituent species following that of PalaearcticRegion. A yearly progress in taxonomic research of black flies is summarized in an e-article“World blackflies (Diptera: Simuliidae): A comprehensive revision of the Taxonomic andGeographical Inventory” annually revised and issued by P. H. Adler & R. W. Crosskey,which is an excellent and significant compilation providing an up-to-date information of theclassification of black flies around the world. Based on its latest version (2016), theOriental fauna of black flies is composed of 524 species (23.8% of the total world numberof extant species, 2,204), which are classified in 10 subgenera of the genus Simulium, i.e.,three endemic subgenera (Asiosimulium, Daviesellum, and Wallacellum), one semi-endemic(Gomphostilbia), two cosmopolitan (Nevermannia and Simulium), and four of a Holarcticdistribution (Byssodon, Eusimulium, Montisimulium, and Wilhelmia). In the OrientalRegion, the subgenus Simulium is dominant, having 232 species (44.3%), followed byGomphostilbia with 193 component species (36.8%) and Nevermannia with 55 species(10.5%), while seven other subgenera consist of 1–16 species. The Oriental simuliid faunaso far unveiled is characterized by a wealth of diversities in morphological characters(including several novel characters) and phylogenetic lineages (in term of the number of thesubgenera and species-groups), which are valued to be equal to or exceed those in otherregions. On the other hand, the lack of primitive supraspecific taxa has no equal, andvarious continental and insular geographical distribution patterns displayed by differentlineages, that might have been caused by their different abilities of adaptation to theclimatic changes in the cyclic ice ages under the different geological, geographical and The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

6ecological conditions in the Oriental Region, are also striking. In this presentation,highlighted are: 1) Brief introduction of the simuliid fauna of the Oriental Region,compared with those of five other zoogeographical regions; 2) Speciation strategiesspeculated by morphological changes of pupal gills; 3) geographical dispersals of certainlineages based on a hypothesis that changes occur from plesiomorphic to apomorphicmorphological characters. Finally, current taxonomic problems, in particular, over certainold species, are noted, and needs for future black fly studies including species complex,intra- and inter-lineage relationships, origins of endemic and semi-endemic subgenera,biological aspects of each taxon (e.g., biting habits, life history, interactions withenvironments, natural enemies), roles in the transmission of pathogens and parasites, androles in the food chain (or energy circulation) of the aquatic ecosystem, and extinction ofspecies, are emphasized.Keywords: black fly, Simuliidae, biodiversity, fauna, Oriental region*Corresponding author: Hiroyuki Takaoka E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

7 Assemblage structure of larval black fly (Simuliidae: Diptera) species in South India Sankarappan Anbalagan* Department of Zoology, Sethupathy Government Arts College (Affiliated to Alagappa University), Ramanathapuram-623502, Tamil Nadu, India ABSTRACT The assemblage structure of larval black fly species (Simuliidae: Diptera) in 24 streamsfrom three biosphere reserves in South India was investigated. A total of 16 species and 2subgenera (Simulium and Gomphostilbia) of Simulium were collected. Among thesespecies, S. (S.) gravelyi (16%) and S. (S.) striatum (15.1%) were abundantly found and S.(S.) striatum was commonly distributed in many streams. The most of species was notdistributed more than two or three streams. Diversity analyses and degree of pollution usingFamily Biotic Index (FBI) were analyzed. Among environmental variables examinedlatitude, stream order, pH, FBI and conductivity were significantly related to thedistribution of larval black flies revealed by multivariate analyses. Individual test betweenstatistically important environmental variables and larval black fly species showed thatlatitude, stream order and FBI were positively determined the assemblages of larval blackfly species in streams. The present study suggests that latitude and stream order play amajor role for the assemblage of species and anthropogenic impact increases the abundanceof larval black flies.Keywords: diversity, distribution, environmental factors, anthropogenic impact, Simulium*Corresponding author: S. AnbalaganPhone: +91-9360525978E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

8 Cryptic diversity in human-biting black flies (Diptera: Simuliidae) in Thailand Pairot Pramual1,2*, Jiraporn Thaijarern1, Komgrit Wongpakam3 1Department of Biology, Faculty of Science, Mahasarakham University, Kantharawichai District, Maha Sarakham, 44150 Thailand 2Biodiverstiy and Traditional Knowledge Research Unit, Mahasarakham University, Kantharawichai District, Maha Sarakham, 44150 Thailand, E-mail: [email protected] 3Walai Rukhavej Botanical Research Institute, Mahasarakham University, Kantharawichai District, Maha Sarakham, 44150 Thailand, E-mail: [email protected] ABSTRACT Black flies (Diptera: Simuliidae) are important insect vectors and pests for humansand animals. Accurate identification, therefore, is important for control and management. Inthis study, we used mitochondrial cytochrome oxidase I (COI) barcoding sequences to testthe efficiency of species identification for the human-biting black flies in Thailand. Wedirectly used the human-biting specimens because they enable us to link informationbetween different development stages. Three black fly species, Simulium nodosum, S.nigrogilvum and S. doipuiense complex, were collected, and one of the S. doipuiensecomplex was confirmed for the first time as having a human-biting habit. The COIsequences revealed considerable genetic diversity in all three species. Comparisons to aCOI sequence library of black flies in Thailand and in a public database indicate a highefficiency for specimen identification for S. nodosum and S. nigrolvum, but this method isnot successful for the S. doipuiense complex. Phylogenetic analyses revealed two divergentlineages in the S. doipuiense complex. Human-biting specimens formed a separate cladefrom the other members of this species. The results are consistent with the Barcoding IndexNumber System (BINs) analysis that found six BINs in the S. doipuiense complex. Furthertaxonomic work is needed to clarify the species status of these man-biting specimens.Keywords: black fly, DNA barcode, Simulium, Simuliidae, Thailand*Corresponding author: Pairot PramualPhone: 66(0)43-754-245; Fax: 66(0)-43-754-245E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

9 The Biodiversity of Black flies (Diptera: Simuliidae) in Indonesia Upik Kesumawati Hadi1, Hiroyuki Takaoka2 1Entomology Laboratory, Division of Parasitology and Medical Entomology, Department of Animal Infectious Diseases and Veterinary Public Health, Faculty of Veterinary Medicine, Bogor Agricultural University, Jl Agatis Darmaga Bogor 16880, Indonesia 2University of Malaya, Jl Universiti, 50603 Kuala Lumpur, Wilayah Persekutuan Kuala Lumpur, Malaysia ABSTRACT Indonesia is one of the megadiversity country in the world endowed with rich andunique biodiversity insects such as blackflies species ( Diptera: Simuliidae). Blackflies arefound almost anywhere with running water suitable as habitat for the immature stages. Thisfamily is one of the most important groups of blood-sucking insects. This study collates thethe records of Simulium (Diptera: Simuliidae) in previous publications related fauna ofIndonesia. Based on the results of this study, there were 122 species of blackflies inIndonesian Archipelago. All species are assigned to the genus Simulium Latreille s.l., andare placed into five subgenera, i.e. Gomphostilbia Enderlein, Morops Enderlein,Nevermannia Enderlein, Simulium Latreille s.str. and Wallacellum Takaoka. Furtherclassification into 27 species groups within the subgenera were also made. Checklists ofIndonesian Simuliidae are provided including data on the distribution of each species.Keywords: Biodiversity, Diptera, Indonesia, Simuliidae*Corresponding author: Upik Kesumawati Hadi E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

10 Faunas of blackflies and administrative borders S.V. Aibulatov Zoological Institute of the Russian Academy of Sciences, Universitetskaya emb., 1, St.-Petersburg, 199034, Russia ABSTRACT In many countries, including ex-USSR countries, research of blackflies (as well asresearches of other elements of fauna) was undertaken in the conditions of a peculiarisolation within state borders. In this regard, there appeared considerable inequalities ofgeographically close faunae. Possible reasons of such inequalities are listed below: - different scientific traditions, - language isolation, - poor study of frontier territories. The last point (poor study of frontier areas) evokes a strong need for parallelresearch of border territories, as well as for collection exchange between scientificinstitutions of the different countries. The term ‘frontier territories’ implies natural areas located in one river basin (or onone large island or the archipelago), at one height above the sea level, in one natural zone.Keywords: fauna, blackflies*Corresponding author: S.V. AibulatovPhone.: 8-9213659041E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

11 Advances in Simuliidology and Changes in the Geographic Centers of Research Peter H. Adler Clemson University, Department of Agricultural and Environmental Sciences, Clemson, SC 29634-0310 USA ABSTRACTMedical and veterinary problems, including vector-borne disease, historically have drivenmuch of the research on the family Simuliidae. The success of Bti in suppressing simuliidpopulations has reduced research efforts in some areas of the world. Consequently,investment in simuliid research has shifted from geographic areas, such as North America,to other areas, such as Southeast Asia, where new centers of simuliid excellence havegrown. At the same time, new tools, particularly in the molecular area, have openedopportunities to explore questions that, until recently, would have been impossible toaddress. Thus, while the geographic centers of research are changing and new tools,approaches, and capabilities are continuing to develop, it is worth examining examples ofresearch topics that might offer the most productive opportunities to advance ourunderstanding of the family Simuliidae. As a springboard for considering the futuredirection of simuliidology, I offer the following five focal topics: (1) Integrativesystematics: The full integration of morphology, chromosomes, and molecules is still in itsinfancy for the Simuliidae. It offers not only the greatest hope for resolving practicalproblems, such as identification, but also for understanding the deeper questions ofevolutionary biology, such as the driving forces of simuliid diversification. (2) Factorsdetermining selection of oviposition sites: Virtually all ecological studies of black fliesultimately trace back to where the females deposited their eggs. Ironically, we know littleabout the cues used by females to make ovipositional decisions at the various spatial scales.(3) Dispersal distances of adults: A second major factor determining much of the ecologyof black flies, as well as our ability to suppress their populations, is how far the adults,particularly the females, disperse from their natal waters. (4) Symbiote-host relationships:Each individual simuliid is an ecosystem of other organisms, such as fungi and nematodes,as well as dozens, perhaps hundreds of bacterial species, of which we know virtuallynothing, but which undoubtedly play major roles in driving simuliid biology. (5) Wildblood hosts: The vertebrate hosts of most species of black flies are either completely The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

12unknown or incompletely known, limiting not only our ability to manage pest populations,but also to understand basic ecological and evolutionary aspects of simuliids.Keywords: blood hosts, Bti, dispersal, integrative systematics, oviposition, symbiotes*Corresponding author. Peter H. AdlerPhone: 864-656-5044; Fax: 864-656-5069E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

13 Symbionts of Larval black flies (Diptera: Simuliidae) in Thailand Sanae Jitklang1*, Chaliow Kuvangkadilok2, Visut Baimai2, and Peter H. Adler3 1Conservation Biology Program, Division of Biological and Natural Resource Sciences, Mahidol University, Kanchanaburi Campus, Sai Yok District, Kanchanaburi 71150 2Department of Biology, Faculty of Science, Mahidol University, Ratchathewi, Bangkok 10400 3Entomology Program, School of Agricultural, Forest, and Environmental Sciences, Clemson University, Clemson, South Carolina 29634-0310 USA ABSTRACTLarval black flies are a major component of the macroinvertebrate fauna in streams andrivers. They are hosts of various parasites e.g., mermithid nematodes and fungi. In addition,they should be model hosts for investigating the associations of symbiotic organisms becausethey are taxonomically well known at the species level. However, the symbionts of larvalblack flies are almost unknown in Thailand. Therefore, the northern and western regions ofThailand were selected as major sites for sampling in the present study to screen thesymbionts and to determine the prevalence of infection. The study was conducted in theprovinces that consist of mountains as a main source of streams and rivers. The larval blackflies were collected from 27 stream sites and 13 sites in northern and western regions,respectively. The most frequently collected species were Simulium nakhonense and S.quinquestriatum, accounting for 67%. The total of eleven larval species was infected withparasites: S. asakoae, S. chamlongi, S. chiangmaiense, S. fenestratum, S. feuerborni, S.nakhonense, S. nodosum, S. quinquestriatum, S. rudnicki, S. siamense and S. tani. Threegroups of parasites were reported in this study including mermithid nematodes(Mermithidae), microsporidian fungi (Zygomycota) and the fungus Coelomycidium simuliiDebaisieux (Blastocladiomycetes). The trichomycete fungi and ichthyosporean protozoa werefound in the mid and hindgut of larval black flies. Harpella melusinae and Stachylina sp.(Harpellaceae) was found in the midgut of larval black flies. Four species of trichomycetes(Legeriomycetaceae): Genistellospora homothallica, Pennella sp., Simuliomyces microsporusand Smittium sp. including one species of protozoa, Paramoebidium sp. (Mesomycetozoea)were found in the hindguts of host species. H. melusinae was present throughout the year inS. nakhonense larvae with percentages of infection (0-100%). These symbionts were moreprevalent in common black fly species. Environmental factors have played a role in thedistribution of the symbionts such as stream size, velocity and rainfall. We expect that this The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

14study will be well worth discovering and providing a means to attract attention to theimportance of symbiosis in the black flies of Thailand.Keywords: Larval black flies, Symbionts, Mermithidae, Microsporidia, Trichomycetes,Thailand*Corresponding author.Phone: 085-1427-395E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

15 ABSTRACTSPoster Presentation The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

16 Species and Population Diversity of Simulium (Diptera; Simuliidae) in Kupang District of East Nusa Tenggara Julianty Almet1, Upik Kesumawati Hadi2, Susi Soviana2, Supriyono2 1 Faculty of Veterinary Medicine Nusa Cendana University, Kupang East Nusa Tenggara, Indonesia2Division of Parasitology and Medical Entomology, Department of Animal Disease and Veterinary Public Health Faculty of Veterinary Medicine IPB, Indonesia Jl. Agathis kampus IPB Darmaga, Bogor 16680. Indonesia ABSTRACT Black flies (Simuliidae) are one of the most important biting insects due to their blood-sucking habits and ability to transmit parasites. There is no sufficient information on the flyspecies in Indonesia, particularly in relation to taxonomical and ecological aspects.Therefore, the present study was conducted to explore black flies in Indonesia to becompared with other regions. This study aimed to identify Simulium species from Oenesuand Oehala Waterfalls, study the habitat of Simulium larvae, and the density fluctuation ofSimulium fly on large livestock. The study was expected to contribute adding informationon ectoparasite biodiversity in Indonesia, particularly in East Nusa Tenggara Province.The study was conducted at Oenesu of West Kupang District and Oehala of South TimorTengah District. The collections of blackflies larvae were done every weeks, and the adultcollections were done in cattle shew by using light trap at 18:00-06:00 once in two weeksfor three months. The study revealed there were four identified Simulium species fromOenesu site, namely S. (Simulium) timorense (75.59%), S. (Gompostilbia) sundaicum(18.76%), S. (Wallacellum) sp (3.72%), and S. (Nevermania) aureohirtum (10.2%); whilefrom Oehala were 3 Simulium larval species, i.e. Simulium (S.) timorense (46.5%),Simulium (G.) sundaicum (43.1%), and Simulium (N.) aureohirtum (10.2%). Oenesu andOehala study sites were suitable for Simulium larval growth based on the habitatcharacteristic such as temperature, pH, current to velocity, and shade presence. Therewere a total of 2 adult fly species (52 flies) collected from livestock pens, i.e. S (S.)timorense (71.5%), and S (G.) sundaicum (28.8%). The blood-sucking activity of S. (S).timorense was started at 18:00; while S. (G.) sundaicum at 20:00. The peak of blood-sucking activity of the two fly species was at 00:00-02:00.Keywords: Blackfly, East Nusa Tenggara, Oenesu, Oehala, Simulium sp The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

17 Larval morphological polymorphisms in Simulium fenestratum Edwards, 1934 (Diptera: Simuliidae) Jiraporn Thaijarern1, Peter H. Adler2, Pairot Pramual1* 1 Department of Biology, Faculty of Science, Mahasarakham University, Kantharawichai District, Maha Sarakham 44150, Thailand. 2Entomology Program, School of Agricultural, Forest & Environmental Sciences, Clemson University, Clemson, SC 29634-0310, USA ABSTRACTSimulium fenestratum Edwards is a member of the S. multistiatum species group. Thisspecies is geographically and ecologically widespread. Two morphological forms that weredifferentiated by the presence or absence of dorsolateral protuberances were found inThailand. In this study, we used an integrated approach based on cytogenetic and moleculargenetic evidence to test the species status of these morphological forms. A total of 177specimens (136 specimens with protuberances and 41 specimens without protuberances)from 10 sampling sites throughout Thailand were cytological examined. The bandingpatterns of the polytene chromosomes of these morphological forms were not different.DNA sequences of the cytochrome c oxidase I (COI) and elongation complex protein 1(ECP1) were obtained from 27 specimens (12 with protuberances and 15 withoutprotuberances). The maximum genetic distances within the group of larvae withprotuberances were COI (0.032), ECP1 (0.077) and combined dataset (0.075), while for thelarvae without protuberances they were COI (0.048), ECP1 (0.038) and combined dataset(0.068), which had overlapping minimum genetic distances between the two forms (COI =0.003, ECP1 = 0.009, combined dataset = 0.010). Phylogenetic analysis revealed no clearseparation between the two groups. Ecological analysis showed no significant differencesbetween the habitats of these two forms. Therefore, multiple character sets indicated thatthese two morphological forms belong to the same species.Keywords: black flies, Simulium fenestratum, polytene chromosome, morphologicalpolymorphism*Corresponding author: Pairot PramualPhone: 6643754245+ext. 1122; Fax: 66+43754245E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

18 Three taxa in one: cryptic diversity in the black fly Simulium nobile (Diptera: Simuliidae) in Southeast Asia Low, V. L.1*, Takaoka, H.1, Pramual, P. 2 Adler, P. H.3, Ya’cob, Z.1, Chen, C. D.1, Yotopranoto, S.4, Zaid, A. 5, Hadi, U. K.6, Lardizabal, M. L.7, Nasruddin-Roshidi, A.1, Sofian-AzirunM. 1 1Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia 2Department of Biology, Faculty of Science, Mahasarakham University, Maha Sarakham, Thailand 3Department of Agricultural and Environmental Sciences, Clemson University, Clemson, SC, USA 4Laboratory of Entomology, Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia 5University of Technology Yogyakarta, Jalan Ring Road Utara Jombor Sleman, Yogyakarta, Indonesia 6Department of Animal Infectious Diseases and Veterinary Public Health, Faculty of Veterinary Medicine, Bogor Agricultural University, Bogor, Indonesia 7International Tropical Forestry Programme, Faculty of Science and Natural Resources, Universiti Malaysia Sabah, Kota Kinabalu, Sabah, Malaysia ABSTRACTWe access the molecular diversity of the black fly Simulium nobile De Mejiere, using theuniversal cytochrome c oxidase subunit I (COI) barcoding gene, across its distributionalrange in Southeast Asia. Our phylogenetic analyses recovered three well-supportedmitochondrial lineages of S. nobile, suggesting the presence of cryptic species. Lineage A iscomprised of a population from Sabah, East Malaysia (Borneo); lineage B represents thetype population from Java, Indonesia; and lineage C includes populations from themainland of Southeast Asia (Peninsular Malaysia and Thailand). The genetic variation oflineage C on the mainland is greater than that of lineages A and B on the islands of Borneoand Java. Our study highlights the value of a molecular approach in assessing species statusof simuliids in geographically distinct regions.Keywords: Simulium nobile species group, COI gene, phylogenetics, systematics*Corresponding author: V. L. LowPhone: +6016-5605857E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

19 Multiple character sets revealed conspecificity betweenSimulium chiangmaiense and S. nakhonense (Diptera: Simuliidae) Sirinapa Pangjanda1, Chaliow Kuvangkadilok2, Pairot Pramual1* 1Department of Biology, Faculty of Science, Mahasarakham University, Kantharawichai District, Maha Sarakham 44150, Thailand 2Department of Biology, Faculty of Science, Mahidol University, Bangkok 10400 Thailand ABSTRACTA total of four black fly species of the Simulium striatum species group have been recordedin Thailand. The morphological characteristics of the larval stages of these species arehighly similar. In this study, we used molecular genetic markers based on cytochrome coxidase subunit I (COI) and elongation complex protein 1 (ECP1) sequences todifferentiate the larvae of S. chiangmaiense, S. quinquestiatum and S. nakhonense. Theresults revealed that S. chiangmaiense and S. nakhonense were not genetically different,while S. quinquestiatum was clearly separated. S. chiangmaiense and S. nakhonense arealso similar in the larval habitat characteristics and have undifferentiated polytenechromosome banding patterns. These species are only separable morphologically by thenumber of gill filaments. Therefore, S. chiangmaiense and S. nakhonense are possibly thesame biological species that is polymorphic for the number of gill filaments.Keywords: Simuliidae, black flies, Simulium striatum species group*Corresponding author: Pairot PramualPhone: 66(0)43-754-245; Fax: 66(0)-43-754-245E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

20Distribution pattern of black fly (Diptera: Simuliidae) assemblages along an altitudinal gradient in Peninsular Malaysia Zubaidah Ya’cob1*, Hiroyuki Takaoka1, Pairot Pramual2, Van Lun Low1, Mohd Sofian-Azirun1* 1Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia 2Department of Biology, Faculty of Science, Mahasarakham University, Maha Sarakham 44150, Thailand ABSTRACTPreimaginal black flies (Diptera: Simuliidae) are important components of the streamecosystem. However, there has been limited research undertaken on the vertical distributionof preimaginal black flies and their associated ecological factors. Stream conditions aregenerally variable along the altitudinal gradient. Therefore, we conducted an in-depthentomological survey to investigate the simuliid distribution pattern along an altitudinalgradient in Peninsular Malaysia. A total of 432 collections were performed in this study (24samplings at each of 18 fixed-streams at monthly intervals) from February 2012 to January2014. Larvae and pupae attached on aquatic substrates such as grasses, leaves and stems,twigs, plant roots and rocks were collected by hand using fine forceps. Stream depth (m),width (m), velocity (m/s), water temperature (°C), acidity (pH), conductivity (mS/cm) anddissolved oxygen (mg/L) were measured at the time of each collection. A total of 35 blackfly species were recorded in the present study. The most frequently collected species wereSimulium tani (31.7 %) and S. whartoni (21.5 %), while the relatively common specieswere Simulium sp. (nr. feuerborni) (16.2 %), S. decuplum (15.5 %), S. angulistylum(14.8 %), S. bishopi (13.2 %) and S. izuae (11.8 %). Total estimated species richnessranged between 39.8 and 41.3, which yielded more than 80 % of sampling efficiency. Sixsimuliid species were distributed below 500 m, whereas eight species were distributedabove 1400 m. Simulium sp. (nr. feuerborni) and S. asakoae were found from middle tohigh altitudes (711–1813 m). Simulium whartoni, S. brevipar and S. bishopi weredistributed widely from low to high altitudes (159–1813 m). Regression analysis betweenspecies richness and PCs revealed that the species richness was significantly associatedwith wider, deeper and faster streams at low altitude, normal water temperature (23–25 °C),low conductivity, higher discharge, more canopy cover and riparian vegetation and with The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

21larger streambed particles (F = 20.8, df = 1, 422, P < 0.001). Forward logistic regressionindicated four species were significantly related to the stream variables (S. whartoni,Simulium sp. (nr. feuerborni), S. tani and S. angulistylum). Canonical correspondenceanalysis indicated that the temperature, stream size and discharge were the most importantfactors contributing to the separation of the stream sites from different altitude and henceare the predictors for the distribution of black fly species assemblages. This study hasprovided insight into the distribution pattern of preimaginal black fly assemblages along analtitudinal gradient in Peninsular Malaysia. This study could deepen our knowledge on theecology and biology of the specialised taxa in response to environmental changes.Keywords: Black fly, Simulium, Elevation, Habitat characteristics, Peninsular Malaysia.*Corresponding author: Zubaidah Ya’cobPhone: 012-64347071E-mail: [email protected];*Corresponding author: Mohd Sofian-AzirunE-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

22Lists of Symposium Participants The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

23Peter H. Adler, Department of Agricultural and Environmental Sciences, Clemson University, Clemson, SC 29634-0310 USA, E-mail: [email protected] V. Aibulatov, Zoological Institute of the Russian Academy of Sciences, Universitetskaya emb., 1, St.-Petersburg, 199034, Russia E-mail: [email protected] Almet, Faculty of Veterinary Medicine, Nusa Cendana University, Kupang East Nusa Tenggara, IndonesiaSankarappan Anbalagan, Department of Zoology, Sethupathy Government Arts College (Affiliated to Alagappa University), Ramanathapuram-623502, Tamil Nadu, India E-mail: [email protected] Baimai, Department of Biology, Faculty of Science, Mahidol University, Ratchathewi, Bangkok 10400 Thailand E-mail: [email protected] Dhang Chen, Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia E-mail: [email protected] Kesumawati Hadi, Division of Parasitology and Medical Entomology, Department of Animal Disease and Veterinary Public Health Faculty of Veterinary Medicine IPB, Bogor Agricultural University, Jl. Agathis kampus IPB Darmaga, Bogor 16680 Indonesia E-mail: [email protected] Jaihan, Department of Biology, Faculty of Science, Mahasarakham University, Kantharawichai District, Maha Sarakham, 44150 Thailand E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

24Sanae Jitklang, Conservation Biology Program, Division of Biological and Natural Resource Sciences, Mahidol University, Kanchanaburi Campus, Sai Yok District, Kanchanaburi 71150 Thailand E-mail: [email protected] Kunprom, Department of Biology, Faculty of Science, Mahasarakham University, Kantharawichai District, Maha Sarakham, 44150 Thailand E-mail: [email protected] Kuvangkadilok, Department of Biology, Faculty of Science, Mahidol University, Ratchathewi, Bangkok 10400 E-mail: [email protected] Lourdes Lardizabal, International Tropical Forestry Programme, Faculty of Science and Natural Resources, Universiti Malaysia Sabah, Kota Kinabalu, Sabah, Malaysia E-mail: [email protected] Lun Low, Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia E-mail: [email protected] Nasruddin-Roshidi, Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia E-mail: [email protected] Pangjanda, Department of Biology, Faculty of Science, Mahasarakham University, Kantharawichai District, Maha Sarakham, 44150 Thailand E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

Pairot Pramual, 25Mohd Sofian-Azirun,Susi Soviana, Department of Biology, Faculty of Science,Wichai Srisuka, Mahasarakham University, Kantharawichai District,Supriyono, Maha Sarakham, 44150 ThailandHiroyuki Takaoka, E-mail: [email protected] Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia E-mail: [email protected] Division of Parasitology and Medical Entomology, Department of Animal Disease and Veterinary Public Health Faculty of Veterinary Medicine IPB, Bogor Agricultural University, Jl. Agathis kampus IPB Darmaga, Bogor 16680. Indonesia Queen Sirikit Botanic Garden, PO Box7, Maerim- Samoeng Road, Maerim District, Chiang Mai, 50180 Thailand E-mail: [email protected] Division of Parasitology and Medical Entomology, Department of Animal Disease and Veterinary Public Health Faculty of Veterinary Medicine IPB, Bogor Agricultural University, Jl. Agathis kampus IPB Darmaga, Bogor 16680. Indonesia Institute of Biological Sciences, Faculty of Science, University of Malaya, Malaysia; Professor Emeritus, Oita University, Japan; Distinguished fellow, The Japanese Society of Tropical Medicine, Japan E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand

26Ubon Tangkawanit, Department of Entomology, Faculty of Agriculture,Jiraporn Thaijarern, Khon Kaen University, Khon Kaen 40002 ThailandKomgrit Wongpakam, E-mail: [email protected] Ya’cob,Subagyo Yotopranoto, Department of Biology, Faculty of Science, MahasarakhamAdnan Zaid, University, Kantharawichai District, Maha Sarakham, 44150 Thailand E-mail: [email protected] Walai Rukhavej Botanical Research Institute, Mahasarakham University, Kantharawichai District, Maha Sarakham, 44150 Thailand, E-mail: [email protected] Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia E-mail: [email protected] Laboratory of Entomology, Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia E-mail: [email protected] University of Technology Yogyakarta, Jalan Ring Road Utara Jombor Sleman, Yogyakarta, Indonesia E-mail: [email protected] The 1st Asian Simuliidae Symposium, 25 – 27 July 2016, Mahasarakham University, Maha Sarakham, Thailand