Morphology, molecular identification, and pathogenicity of Vibrio spp. on blood clam (Anadara granosa) in Yogyakarta, Indonesia tourism beach areas
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Abstract. Devi AR, Susilowati A, Setyaningsih R. 2019. Morphology, molecular identification, and pathogenicity of Vibrio spp. on blood clam (Anadara granosa) in Yogyakarta, Indonesia tourism beach areas. Biodiversitas 20: 2890-2896. Seafood is very popular among Indonesian people, especially in coastal areas. In Bantul Yogyakarta, blood clams have become one of tourist's favorite, either cooked or raw. Blood clams are filter feeders that cause the clams to be vulnerable to contamination of pathogenic bacteria that can cause foodborne disease, including Escherichia, Pseudomonas, and Vibrio. The 10-20% cases of foodborne disease transmitted through seafood caused by Vibrio spp. Three species of Vibrio can cause foodborne disease in humans, i.e., V. cholerae, V. parahaemolyticus, and V. vulnificus. The purpose of this study was to determine the character of Vibrio using morphological and molecular identification and pathogenicity on blood clam (Anadara granosa). Blood clams samples were collected from Depok, Goa Cemara, and Kwaru beaches in Bantul, Yogyakarta, Indonesia. Isolation of Vibrio spp. from blood clams was done using selective differential Thiosulfate Citrate bile salts sucrose (TCBS) culture medium. The morphological characters of the isolate colonies were determined based on the color, shape, texture, and size of the colony. Hemolysis test was also performed to evaluate the pathogenicity by using blood agar media. Molecular identification of Vibrio species was made using 16S rRNA sequence. Phylogenetic analysis was performed using Neighbor-Joining method in Mega X software. Samples for the analysis came from DNA sequences of this study and those from the GenBank database. Of the total 15 isolates obtained, four isolates showed positive ?-hemolysis, namely, isolate P2S2 -1bH, P3S1-1aH, P2S1-1aK and P2S2-1aK, and one isolate had positive ?-hemolysis (P3S2-1aK). Seven species of Vibrio were identified as V. algynolyticus, V. parahaemolyticus, V. diabolicus, V neocaledonicus, V. azureus, V. natrigens, and V. cholerae.
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References
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Anses. 2012. Vibrio parahaemolyticus Family Vibrionaceae, genus Vibrio Bacterium.Data Sheet on Foodborne disease Biological Hazard.
Azwai SM., Alfallani EA., Abolghait SK. Garbaj. 2016. Isolation and Molecular Identification of Vibrio spp by sequencing of 16S rDNA from Seafood, Meat and Meat Products in Libya. Open Veterinary Journal Vol.6 (1): 36-43. Tripoli, Libya.
Belkin, S. And Colwell RR. 2005. Ocean and Health: Pathogens in the Marine Environment. Springer. Newyork.
Ben Kahla-Nakbi A., Besbes A., Chaieb K., Rouabhia M., Bakhrouf A. 2007. Survival of Vibrio alginolyticus in Seawater and Retention of Virulence of its Straved Cells. Elsevier Marine Environmental Research 64 469-478.
Campanelli A., Sanchez-Pollita S., Saurat J.H. 2008 Ulceration cutanee apres morsure de poulpe: Infection a Vibrio alginolyticus un pathogene emergent. Annales de Dermatologie et de Venerologie 135:225-227.
Duangkhae, K. Pimpa B., and Chowpongpang S. 2011. Occurrence of Vibrio parahaemolytcus in cockle (Anadara granosa) harvested from the South Coast of Thailand. Songklanakarin Journal of Science And Technology (SJST) 33(3) 295-300. Thailand.
Dog-Yong L., Soo-Yun M, Sang-Oh L., Hee-Young., Hee-Joo L., Mi Suk. 2008. Septic Shock due to Vibrio alginolytcus in a Cirrhotic Patient: The First Case in Korea. Yonsei Med Journal 49:329-332.
Ennaji M., Cohen N. 2013. Vibrio Alginolyticus: An Emerging Pathogen of Foodborne Disease. IJST (International Journal of Science and Technology) V.2/4. Casabanca Morocco.
Fajriani B., Budiharjo A., dan Pujiyanto S. 2018. Isolasi dan Identifikasi Molekuler Bakteri Antagonis Terhadap Vibrio parahaemolyticus Patogen pada Udang Litopenaeus vannamei dari Produk Probiotik dan Sedimen Mangrove di Rembang. Jurnal Biologi Volume 7 (1) Hal 52-53
Fatchiyah A. Widyarti E., Rahayu S. 2011. Biologi Molekuler Prinsip Dasar Analisis. Jakarta: Erlangga.
Felix F., Nogroho TT., Silalahi S., Oktavia Y. 2011. Screening of Indonesian Original bacterial Vibrio sp As A Cause of Shrimp Diseases Based on 16S Ribosomal DNA-Technique. Tropical Marine Science and Technology Journal Vol 3/2 85-99.
Hidayat, ARS. 2013. Karakaterisasi Bakteri Genus Vibrio dari Ikan Kerapu (Plectropomus sp.). Jurnal Biogenesis ISSN 2302-1616 1(2): 141-143.
Ihsan B., Retnaringrum E. 2017. Isolasi dan Identifikasi Bakteri Vibrio sp pada Kerang Kapah (Meretrix meretrix) di Kabupaten Trenggalek. Borneo Harpodon Journal V 10/1.
Ilmiah, Sukenda, Widanarni dan Enang, H. 2012. Isolation and characterization of Pathogenic Vibrio on Tiger grouper Epinephelus fuscoguttatus. Journal Akuakultur Indonesia 11 (1), 28-37. Bogor.
Jiang Yanhua, Chu Y., Xie G., et al. 2019. Antimicrobial resistance, virulence and Genetic Relationship of Vibrio parahaemolyticus in seafood from coast Bohai sea and Yellow Sea, China. Int Journal of Food Microbiology. 290: 116-124.
Johnson M, Zaretskaya I., Raytselis Y, Marezhuk, Y, McGinnis., and Madden TL. 2008. NCBI BLAST: A Better Web Interface. Nucleic Acid Research. 36(2) 5-9
Mailoa, MN., Setha B. 2011. Karakteristik Patogenitas Vibrio sp. Diisolasi dari Lendir Sidat (Anguilla sp) Jurnal Kedokteran dan Kesehatan. Ambon.
Moradi I., Song Z., Tao X,. 2015. Introducing a Novel Bacterium, Vibrio neocaledonicus sp with The Highest Corrosion Inhibition Efficiency. Electrochemistry Communication Journal 64-68. China.
Mudatsir. 2007. Faktor-Faktor yang Mempengaruhi Kehidupan Mikroba dalam Air. Medicine Syiah Kuala Journal Vol7/1.
Nguyen Nam-Phuong, Tandy W., Mihai P and Bryan W. 2016. A Perspective on 16S rRNA Operational Taxonomic Unitclustering Using Sequence Similarity. Nature Partner Journal. 2: 1-8.
Ningsih HU., Prakarsa TBP., Margawati ET. Koleksi DNA dan Konfirmasi Marka ETH10 Pengkode Sifat Pertumbuhan pad Sapi Pasundan. BIOTROPIC The Journal of Tropical Biology Vol 1 No.1. Cibinong, Bogor.
Nurjanah, Zulhamsyah dan Kustariyah. 2005. Kandungan mineral dan Proksimat Kerang Darah (Anadara granosa) yang diambil dari kabupaten boalemo, Gorontalo. Buletin teknologi Hasil Perikanan 8 (2): 1-4.
Raghunath p. 2015. Roles of Thermostable Direct Hemolysin (TDH) and TDH-related Hemolysin (TRH) in Vibrio parahaemolyticus: Frontiers in Microbiology. 5:8-11
Rossi-Tamisier Morgane, Samia B., Didier R., and Pierre-E. 2015. Cautionary tale of Using 16S rRNA Gene Sequence Similarity Values in Identification of Human-associated Bacterial Species. International Journal of Systematic and Evolutionary Microbiology. 65: 1929-1467.
Schmidt, R.H. Goodrich R.M., Archer D.L dan Schiender, K.R. 2003. General Overview of The Causative Agents of Foodborne Illness. Institute of Food and Agriculture Sciences. University of Florida, USA.
Stephanie and Diana Elisabeth W. 2011. Distribution of INA Bacteria from Rain-Water and Air. Hayati Jurnal of Bioscience. 18: 108-112.
Sujeewa AKW., Norrakiah AS., and Laina M. 2009. Prevalence of Toxic genes of Vibrio parahaemolyticus in shrimps (Penaeus monodon) and culture environment. Int Food Research Journal 16, 89-95.
Turner J.W., Tallman JJ., Macias A, et al. 2018. Comparative Genomic Analysis of Vibrio diabolicus and Six Taxonomic Synonyms: A First Look at The Distribution and Diversity of The Expanded Species. Frontiers in Microbiology Journal DOI: 10.3389.
Widowati R. 2008. Keberadaan bakteri Vibrio parahaemolyticus pada udang yang dijual di Rumah Makan Kawasan Panta Pangandaran. Jurnal Vis Vitalis 1 (1):
Yanez R., Bastias R., Higuera G., Salgado O., Katharios P., Romero., Espejo R., and Garcia K. 2015. Amplification of tlh gene in Other Vibrioanaceae Species by Species-Specific Multiplex PCR of Vibrio parahaemolyticus. Electronic Journal of Biotechnology. 18: 459-463.
Yennie Y., et al. 2015. Prevalence of TDH and TRH Genes of Vibrio parahaemolyticus in Shrimp (Litopenaeus vannamei) from Indramayu, West Java. JPB marine and Fisheries Journal Vol 10/1.
Yoshizawa S., Wada M., Tsukamoto KK, Ikemoto E. A Yakota, Kogure K. 2009. Vibrio azureus sp. nov., a luminous Marine Bacterium Isolated from Seawater. International Journal Syst Evol Microbiol 59(Pt7):1645-9 DOI: 10.1099.
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