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SURVEY OF COLEOPTERANS IN BANGLADESH BY DNA BARCODING OF MALAISE TRAP COLLECTION

Santosh Mazumdar*, Paul D. N. Hebert** and Badrul Amin Bhuiya***

* Department of Zoology, University of Chittagong, BANGLADESH. E-mail: [email protected], ORCID ID: 0000-0001-6403-577X ** Centre for Biodiversity Genomics, University of Guelph, 50 Stone Road East, Guelph, Ontario, CANADA. ORCID ID: 0000-0002-3081-6700 *** Biodiversity Research for Environment & Ecosystem Protection (BREEP), Chattogram- 4325, BANGLADESH.

[Mazumdar, S., Hebert, P. D. N. & Bhuiya, B. A. 2021. Survey of Coleopterans in Bangladesh by DNA barcoding of malaise trap collection. Munis Entomology & Zoology, 16 (1): 275-282]

ABSTRACT: Coleopterans (Insecta: Coleoptera) are the most diverse, species-rich and major ecosystem service providers group of . In the present study, the main objective behind is to confirm identification of coleopterans by using DNA barcoding technique (658 bp sequence from the 5′-end of cytochromeoxidase I) Specimens were collected by a Malaise trap from Chittagong university campus at a site in Bangladesh between April 2014 and March 2015. A total of 14 species, 17 genera, 16 subfamilies and 40 families have been listed in the present study. Of them 6 species viz. Oulema melanopus, Monolepta quadriguttata, Phyllotreta striolata (Chrysomelidae), constricta (), Typhaea stercorea and Carpophilus obsoletus (Mycetophagidae), 7 genera, 4 subfamilies and 16 families are the first country records. This study demonstrates that Order Coleoptera is more diverse group in Bangladesh, and highlighted the need for further investigation in terms of molecular in different areas of the country.

KEY WORDS: Coleoptera, Malaise trap, DNA Barcode, Bangladesh

Order Coleoptera is the largest insect group, containing approximately 360,000 species of named insect species, and occupied 40 percent of total insect world. They are found in about all climates and latitudes (Gressitt, 2018). They play important roles in nearly all terrestrial and freshwater ecosystems (Zhang et al., 2018). Their ecomorphological features that are comparable in a wide range within this group, contributing to overcome difficulties of the taxonomic approach and understand the functioning of ecosystems (Pompeo et al., 2017). In modern biological taxonomic study, DNA barcoding has the potential to mitigate the challenges of accurate identification has become a global research effort since being proposed by Hebert et al. (2003) as a standardized approach for identifying unknown specimens to species-level (Pentinsaari et al., 2019). For evaluating terrestrial insect diversity, DNA barcoding of collected by Malaise traps has captured the headlines (deWaard et al., 2019; Mazumdar et al., 2019). Combination of Malaise traps and pitfall a trapping technique were most effective for families Carabidae, Cerambycidae, and Curculionoidea (Skvarla & Dowling, 2017). In Bangladesh, unlike other insect species, limited molecular studies have been undertaken in the members of Order Coleoptera. Since mid sixties of the last

Munis Entomology & Zoology Mun. Ent. Zool. 276 https://www.munisentzool.org/ (January, 2021) ISSN 1306-3022 © MRG ______century, morphological taxonomic works on coleopterans has quickly gained popularity. For example Alam (1971) reported on coleopterans of stored grains and other stored products. Ahmad & Hossain (1979) noted on weevils (Curculionidae) of the Bangladesh Agricultural University area, Mymensingh. Khan & Khan (2001) studied infestation in dry fish storage. Rahman et al. (1983) worked on coleopteran pest diversity on soybean. Azim (1985) reported coleopteran pests of mango and mango tree. Baksha (1990) reported coleopteran forest insect pest. Ahad et al. (2012) studied aquatic coleopteran Biodiversity of Northern Region of the country. The numbers of DNA barcoded taxonomic works on the species of coleopterans from Bangladesh are very limited. As for example Rain et al. (2016) evaluated Coccinellid predators of aphid and their phylogenetic analysis. Aslam et al. (2019) identified stored grain pest coleopterans. Sarker et al. (2019) identified Coccinella transversalis as agricultural importance. The purpose of this study was to confirm coleopteran diversity through DNA barcoding technique of Malaise trap collection, and at forming a baseline for further studies on this important group of insects in Bangladesh.

MATERIAL AND METHODS

Specimen Collection, Processing, Identification and Specimen Deposition. By following the Standard Operating Protocol for the Global Malaise Trap Program (www.dnabarcoding.ca. ), specimens were collected by a Townes-style Malaise trap (BioQuip Inc. USA) installed in perceived flight paths at Chittagong University Campus (Lat. 22.46359°N; Long. 91.7808°E) in Bangladesh. Between March 2014 and February 2015, the samples were harvested weekly in a 500 mL plastic Nalgene bottle that was filled with 375 mL of 95% ethanol and placed in 500 mL of fresh ethanol before storage at -20°C until analysis. Collected insects were analyzed, following standard barcoding protocols (http://ccdb.ca/resources. php), at the Canadian Centre for DNA Barcoding within the Centre for Biodiversity Genomics, University of Guelph, Canada. Specimen data along with the collection information are accessible in the BOLD (http://v3.boldsystems.org/ index.php/TaxBrowser_Taxonpage ?taxid=125). All the specimens analyzed in this study have been curated at the Centre for Biodiversity Genomics, University of Guelph, Guelph, Ontario, Canada.

Molecular Analysis and Data Analysis. A single leg was removed from each specimen and vouchers were recovered after DNA extraction for imaging and curation. Tissue lysis, DNA extraction, PCR amplification, cycle sequencing and sequence analysis were performed atthe Canadian Centre for DNA Barcoding following the standard protocols (CCDB). PCR amplification of COI-5′ was performed with primers C_LepFolF and C_LepFolR (http://www.ccdb.ca/docs/CCDB_PrimerSets). Following PCR conditions; 94°C (1 min), 5 cycles at 94°C (40 s), 45°C (40 s), 72°C (1 min); 35 cycles at 94°C (40 s), 51°C (40 s), 72°C (1 min) and a final extension at 72°C (5 min) and amplicons were sequenced using BigDye v3.1 (Applied Biosystems) on an ABI 3730XL. Sequences were assembled, aligned, and edited using CodonCode Aligner (CodonCode Corporation, USA) and submitted to Barcode of Life Data Systems (BOLD) (www.boldsystems.org). With a few exceptions, by considering

Munis Entomology & Zoology Mun. Ent. Zool. https://www.munisentzool.org/ (January, 2021) 277 ISSN 1306-3022 © MRG ______sequence matches to records on BOLD, the specimens with barcodes were assigned to 40 families.

RESULTS AND DISCUSSION

Table 1 shows the list of the confirmed species of dipterans in the present study. A total of 14 species, 17 genera, 16 subfamilies and 40 families have been listed in the present study. Of them 6 species viz. Oulema melanopus, Monolepta quadriguttata, Phyllotreta striolata (Chrysomelidae), Cartodere constricta (Latridiidae), Typhaea stercorea and Carpophilus obsoletus (Mycetophagidae), 7 genera, 4 subfamilies and 16 families are new country records. No species were confirmed in seventeen families in current work. In Bangladesh, Alam (1967) listed 19 species of Anthribids (Anthribidae) including Araecerus fasciculatus, and a brief note on forest pests belonging family Brenthidae. Rahman et al. (1983) recorded Oberea brevis (Cerambicidae), Epilachna dodecastigma (Coccinellidae), lribolium sp. (Tenebrionidae) as soybean pests. In a compendium, Gapud (1992) noted three species of Bostrychiidae and five species of Buprestidae as pests of stored grains and economically important crops. Also, Casnoidea indica and Ophionea ishii (Carabidae) are predators of Nilaparvata lugens and Sogatella furcifera (Delphacidae) (Gapud, 1992). In addition, he also mentioned that Carpophilus cylindricus, C. hemipterus and C. mutilates are pests of bottle gourd, dried fruits and vegetables, and joar respectively, and C. obsoletus is new country record, its host is unknown as well. Baksha (1990) reported Dihamnus [Acalolepta] cervinus, Hoplocerambyx spinicornis, Glenea spp. (Cerambycidae); Calopepla leayana (Chrysomelidae); Chrysocroa (Chrysochroa), Catoxantha (Chrysochroa) and Belionata (Belionota) spp. (Buprestidae); Sinoxylon, Dinoderus, Lyctus and Heterobostrychus spp. (Bostrichidae); Leucopholis, Holotrichia and Leucopholis, Holotrichia and Anomala as forest insect pest. Sticholotis chittagongi (Coccinellidae), reported as feeding on aphids infesting Ziziphus mauritiana (Rhamnaceae) in the country (Vazirani, 1983). Khan & Khan (2001) identified Dermestes spp. (Dermestidae), Necrobia rufipes (Cleridae) as dry fish beetle pests. Dicladispa armigera (Chrysomelidae) is a major rice pest of country, and its egg parasitoid Trichogramma zahiri Polaszek (Hymenoptera: Trichogrammatidae) is newly described from this country by Polaszek et al. (2002). Ali et al. (2009) reported wheat pests in storage namely rice weevil (Sitophilus oryzae), grain moth (Sitotroga cerealella) and red flour beetle (Tribolium castaneum). Ahad et al. (2012) recorded aquatic coleopteran insects like Water scavenger (Hydrophilidae), Crawling water beetle (Haliplidae), Diving beetles (Dytiscidae), Whirligig beetles (Gyrinidae). Alam et al. (2019) Sitophilus oryzae, S. zeamais (Curculionidae), Araecerus fasciculatus (Anthribidae), Cryptolestes ferrugineus, C. pusillus (Cucujidae), Prostephanus truncates, Rhyzopertha dominica (Bostrichidae), Tribolium sp., T. castaneum, T. confusum, Tenebrio molitor (Tenebrionidae), Zabrotes subfaciatus (Buruchidae), Oryzaephilus surinamensis (Silvanidae) as maize pest. In Chittagong University Campus, Nasiruddin & Shiuli (2017) recorded Aulacophora sp., Aspidomorpha sp., Phyllophaga crinita, Euchlora sp., Caccobius sp., Mylabris cichorii, Epilachna sp., Alaus sp. belonging to five family namely Crysomelidae, Scarabaeidae, Meloidae, Coccinellidae and Elateridae.

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In molecular identification of Bangladeshi coleopteran fauna, total nine ladybird beetles viz, Adalia bipunctata, Coccinella septempunctata, Coccinella transversalis, Cycloneda munda, Harmonia axyridis, Hippodamia convergens, Micraspis discolor, Scymnus nebulosus, Sticholotis sp. were identified as predators of aphid (Rain et al., 2016). COI genes of three stored grain insect pests i.e,, Sitophilus oryzae, Callosobruchus chinensis and Oryzaephilus surinamensis were sequenced by Aslam et al. (2019). Sarker et al. (2019) utilized DNA barcoding technique for Coccinella transversalis. The current work intended to provide a sighting the diversity of Coleoptera in Bangladesh (Table 2). Much remains to be discovered, therefore, there is a need to carry out systematic surveys in the entire country covering all the seasons to get an optimum estimate of coleopteran diversity.

Table 1. Coleopterans revealed by DNA barcoding of Malaise trap samples collected in Bangladesh.

Recorded from Family Subfamily Scientific name Bangladesh New Aderidae - - New Anthicidae - - Araecerus fasciculatus (De Geer, Alam, 1967 Anthribidae Choraginae 1775) New Biphyllidae - - Gapud, 1992 Bostrichidae - - Alam, 1967 Brentidae - - Gapud, 1992 Buprestidae Agrilinae Agrilus sp. New Cantharidae - - Gapud, 1992 Carabidae - - Gapud, 1992 Cerambycidae - - Gapud, 1992 Chrysomelidae Bruchinae - New Criocerinae Oulema melanopus (Linnaeus, 1758) New Eumolpinae Pagria sp. New Galerucinae - Gapud, 1991; Ahmad et al., Aulacophora lewisii Baly, 1886 2009 Monolepta quadriguttata New (Motschulsky, 1860) Phyllotreta striolata (Fabricius, 1803) New New Clambidae - - Gapud, 1992 Cleridae - - Cheilomenes sexmaculata (Fabricius, Ahmad et al., Coccinellidae Coccinellinae 1781) 2009 Coccinella transversalis Fabricius, Ahmad et al., 1781 2009

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Henosepilachna septima (Dieke, Ahmad et al., Epilachninae 1947) 2009 Henosepilachna vigintioctopunctata Ahmad et al., (Fabricius, 1775) 2009 New Corylophidae - - Xylosandrus crassiusculus New Curculionidae Scolytinae (Motschulsky, 1866) Gapud, 1992 Dermestidae - - New Drilidae - - Gapud, 1992 Elateridae Agrypninae - Ahmad et al., Endomychidae - - 2009 Ahmad et al., Erotylidae - - 2009 Cryptolestes pusillus (Schönherr, New Laemophloeidae - 1817) New Lampyridae Luciolinae - New Ototretinae Lamellipalpus sp. New Latridiidae Corticariinae Migneauxia sp. Cartodere constricta (Gyllenhal, Latridiinae 1827) New New Leiodidae - - Gapud, 1992 Meloidae - - New Melyridae - - New Mordellidae - - New Mycetophagidae Mycetophaginae Typhaea stercorea (Linnaeus, 1758) New Nitidulidae Carpophilinae Carpophilus obsoletus Erichson, 1843 New Phalacridae - - New Ptiliidae - - New Ripiphoridae - - Gapud, 1992 Scarabaeidae - - New Scirtidae - - New Silvanidae - - Gapud, 1992 Staphylinidae - - Gapud, 1992 Tenebrionidae - - New Throscidae - - New Zopheridae - -

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Table 2. Summary of the Coleopterans in Bangladesh.

Family Bangladesh Present study genus /species genus /species (*Ref.) Aderidae - - Anthicidae - - Anthribidae 14/19 1/1 Biphyllidae - Bostrichidae 3/3 Brentidae 3/3 Buprestidae 5/5 1/0 Cantharidae Carabidae 87/23 Cerambycidae 23/27 Chrysomelidae 35/50 5/4 Clambidae - Cleridae 1/1 Coccinellidae 7/14 3/4 Corylophidae - Curculionidae 48/94 1/1 Dermestidae 4/8 Drilidae - - Elateridae 3/1 Endomychidae 5/6 Erotylidae 7/9 Laemophloeidae - 1/1 Lampyridae 1! 1/0 Latridiidae 2/1 2/1 Leiodidae - Meloidae 2/4 Melyridae - - Mordellidae - - Mycetophagidae 1/1 1/1 Nitidulidae 1/4 1/1 Phalacridae - - Ptiliidae - - Ripiphoridae - - Scarabaeidae 26/96 Scirtidae - - Silvanidae - - Staphylinidae 1/1 Tenebrionidae 6/9 Throscidae - - Zopheridae - - * Ref. Gapud, 1992; Ahmad et al., 2008 and results of the present study.

ACKNOWLEDGEMENTS

The authors are indebted to colleagues at the Centre for Biodiversity Genomics, University of Guelph, for aid with sequence analysis.

LITERATURE CITED

Ahad, A., Raja, S. I., Khatun, M. & Hossain, M. A. 2012. Aquatic Insect Biodiversity of Northern Region of Bangladesh. International journal of Bioresource and Stress Management, 3 (3): 387-393.

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Ahmad, M. & Hossain, M. B. 1979. Notes on weevils (Curculionidae: Coleoptera) of the Bangladesh Agricultural University area, Mymensingh. Bangladesh Journal of Zoology, 7 (1): 61-66. Ahmad, M., Kabir, S. M. H., Ahmed, A. T. A., Ahmed, Z. U., Begum, Z. N. T., Hassan, M. A. & Khondker, M. (eds.). 2009. Enccyclopedia of Flora and Fauna of Bangladesh, Vol.22. Pterygota (Part). Asiatic Society of Bangladesh, Dhaka. 426 00. Alam, M. J., Ahmed, K. S., Hossen, B., Mozammel, H. & Hoque, A. B. M. Z. 2019. Storage pests of maize and their status in Bangladesh. Journal of Bioscience and Agriculture Research, 20 (02): 1724-1730. Alam, M. Z. 1967. A Report on the Survey of Insect and Mite Fauna of East Pakistan. East Pakistan Agric. Res. Ins. Dacca, 151 pp. Alam, M. Z. 1971. Pest of stored grains and other stored products and their control. Agricultural Information Service, Dhaka. 61 p. Ali, M. Y., Latif, M. A. & Ali, M. 2009. Effect of some containers, chemicals and indigenous materials on incidence of wheat pests in storage. Journal of Agriculture & Rural Development, 107-113. Aslam, A. F. M., Sultana, S., Rain, F. F., Das, S. R., Siddika, A. & Howlader, A. J. 2019. Molecular characterization and identification of three stored grain pests based on mitochondrial cytochrome C oxidase subunit I (COI) gene sequences. Bangladesh Journal of Zoology, 47 (1): 1-11. Azim, I. I. 1985. Insect pests of mango and their control in Bangladesh. In Symposium on the Problems and Prospects of Mango Production in Bangladesh, Dhaka (Bangladesh), 17-18. Baksha, M. W. 1990. Some major forest insect pests of Bangladesh and their control. Some major forest insect pests of Bangladesh and their control., (1): 1-90. deWaard, J. R., Levesque-Beaudin, V., deWaard, S. L., Ivanova, N. V., McKeown, J. T., Miskie, R., Naik, S., Perez, K. H., Ratnasingham, S., Sobel, C. N. & Sones, J. E. 2019. Expedited assessment of terrestrial diversity by coupling Malaise traps with DNA barcoding. Genome, 62 (3): 85-95. Gapud, V. P. 1992. Insect & mite pests of plant crops in Bangladesh & their enemies: a compendium. United States Agency for International Development / Bangladesh Agricultural Research Council/CHECCI & Co. Consulting Inc. Gressitt, J. L. 2018. Coleopteran. Encyclopaedia Britannica, inc. Available from: https://www.britannica.com/animal/beetle Hebert, P. D. N., Ratnasingham, S. & Dewaard, J. R. 2003. Barcoding life: cytochrome coxidase subunit 1 divergences among closely related species. Proceedings of the Royal Society B: Biological Sciences, 270: 96-99. Khan, M. A. A. & Khan, Y. S. A. 2001. Insect infestation and preventive measures in dry fish storage of Chittagong, Bangladesh. International Journal of Biological Sciences, 1 (10): 963-965. Mazumdar, S., Hebert, P. D. N, Bhuiya, B. A. & Miah, M. I. 2019. Parasitic Hymenoptera Recovered by DNA Barcoding of Malaise Trap Collection at the Chittagong University Campus, Bangladesh. American Journal of BioScience. 7 (6): 94-98. Nasiruddin, M. & Shiuli, F. A. 2017. Entomofauna of four spots of Chittagong University campus and their relative abundance and diversity. Journal of Biodiversity Conservation and Bioresource Management, 3 (1): 55-64. Pentinsaari, M., Anderson, R., Borowiec, L., Bouchard, P., Brunke, A., Douglas, H., Smith, A. B. & Hebert, P. D. 2019. DNA barcodes reveal 63 overlooked species of Canadian beetles (Insecta, Coleoptera). ZooKeys, 894: 53. Polaszek, A., Rabbi, M. F., Islam, Z. & Buckley, Y. M. 2002. Trichogramma zahiri (Hymenoptera: Trichogrammatidae) an egg parasitoid of the rice hispa Dicladispa armigera (Coleoptera: Chrysomelidae) in Bangladesh. Bulletin of Entomological Research, 92: 529-537. Pompeo, P. N., Oliveira Filho, L. C. I. D., Santos, M. A. B. D., Mafra, Á. L., Klauberg Filho, O. & Baretta, D. 2017. 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Plate I. 1. Araecerus fasciculatus (De Geer, 1775), 2. Oulema melanopus (Linnaeus, 1758), 3. Aulacophora lewisii Baly, 1886, 4. Monolepta quadriguttata (Motschulsky, 1860), 5. Phyllotreta striolata (Fabricius, 1803), 6. Cheilomenes sexmaculata (Fabricius, 1781), 7. Coccinella transversalis Fabricius, 1781, 8. Henosepilachna septima (Dieke, 1947), 9. Henosepilachna vigintioctopunctata (Fabricius, 1775), 10. Xylosandrus crassiusculus (Motschulsky, 1866), 11. Cryptolestes pusillus (Schönherr, 1817), 12. Cartodere constricta (Gyllenhal, 1827), 13. Typhaea stercorea (Linnaeus, 1758), 14. Carpophilus obsoletus Erichson, 1843.