Antimicrobial Resistance Profiles and Tetracycline Resistance Genes of Escherichia coli in Mediterranean Mussel and Sea Snails Collected from Black Sea, Turkey

Öz

Aquatic environments are often affected by and exposed to anthropogenic pollutants including antimicrobials used as disease prevention and feed additives. Antimicrobial resistance is a major problem both in animal and in human health worldwide. In this study, Mediterranean mussel (Mytilus galloprovincialis) and Sea snail (Rapana venosa) samples were collected seasonally from the coastline of Black Sea. A total of 54 Escherichia coli were isolated from Mediterranean mussel and Sea snail collected from the coast of Artvin, Rize, Trabzon and Giresun, Turkey. Antimicrobial resistance and the presence of tetracycline (tet) resistance genes (tetA, tetB, tetC, tetD, and tetE) in E. coli isolates were investigated. Antimicrobial susceptibility test determined that 83.3 % of the isolates exhibited resistance to sulfamethoxazole. Resistance to ampicillin and aztreonam was as 66.7 % and 37.0 % among the tested antimicrobials, respectively. The lowest resistant antimicrobial was florfenicol (1.9 %). tetC resistance gene was detected in more than 50% of the isolates. Among the tet resistance genes, tetC was found in the most common gene followed by tetB, tetA, tetE, and tetD. At least one tet gene was detected in 88% of the isolates, and 46% of the isolates had two or more tet genes. The presence of tet resistance genes in E. coli in aquatic environments indicates that these isolates may be a reservoir of tet resistance genes. They may also exhibit an important role in the spread of genes among the pathogenic and non-pathogenic bacteria.

Anahtar Kelimeler

• Antimicrobial,Mytilus galloprovincialis,Rapana venosa,Tetracycline resistance gene

Öz

Doğu Karadeniz (Türkiye)’den Toplanan Kara Midye ve Deniz Salyangozlarındaki Escherichia coli’lerin Antimikrobiyal Direnç Profilleri ve Tetrasiklin Direnç Genleri

Sucul ortamlar sık sık hastalıklardan korunma ve besin katkı maddesi olarak kullanılan antimikrobiyalleri de içeren karasal kökenli kirleticilere maruz kalmakta ve etkilenmektedir. Antimikrobiyal direnç dünya çapında hem insan sağlığında hem de hayvan sağlığı açısından büyük bir problemdir. Bu çalışmada, Karadeniz kıyılarından mevsimlik olarak kara midye ve deniz salyangozu örnekleri toplanmıştır. Artvin, Rize, Trabzon ve Giresun kıyılarından toplanan kara midye ve deniz salyangozlarında toplam 54 adet Escherichia coli suşu izole edilmiştir. E. coli izolatlarında antimikrobiyal direnç ve tetrasiklin direnç genlerinin (tetA, tetB, tetC, tetD, ve tetE) varlığı araştırılmıştır. Antimikrobiyal hassasiyet testi izolatların %83,3’ünün sulfametaksazola karşı direnç olduğunu göstermiştir. Ampisilin ve aztreonama karşı direnç de sırasıyla %66,7 ve %37,0 olarak hesaplanmıştır. En düşük direncin florfenikole karşı olduğu tespit edilmiştir. İzolatların % 50’sinden fazlasında tetC geni bulunmuştur. tet genleri arasında, tetC en çok bulunan gen olurken bunu tetB, tetA, tetE ve tetD takip etmiştir. İzolatların %88’inde en az bir tane test edilen direnç geni olduğu tespit edilmiş ve %46’sı ise iki veya daha fazla tet genine sahip olduğu belirlenmiştir. Sucul ortamlardaki E. coli’lerde tet direnç genlerinin varlığı bu izolatların tet direnç genleri bakımından bir rezervuar olabileceğini göstermektedir. Bu izolatlar ayrıca patojenik ve patojenik olmayan bakteriler arasında da bu genlerin yayılmasında önemli bir rol oynayabilir. 

Anahtar Kelimeler

• Antimikrobiyal,Mytilus galloprovincialis,Rapana venosa,Tetrasiklin direnç geni

Kaynakça

1. Akkan, T., and Mutlu, C., 2016. Determination of Antibiotics Resistance Levels in Enterobacteriaceae Isolated from Giresun Coasts. Turkish Journal of Agriculture-Food Science and Technology, 4(8): 640-650
2. Alkan, N., Aktas, M., and Gedik, K., 2012. Comparison of Metal Accumulation in Fish Species from the Southeastern Black Sea. Bulletin of Environmental Contamination and Toxicology, 88(6): 807-812
3. Altuğ, G., and Güler, N., 2002. Determination of the levels of indicator bacteria, Salmonella spp. and heavy metals in sea snails (Rapana venosa) from the Northern Marmara Sea, Turkey. Turkish Journal of Fisheries and Aquatic Sciences, 2(2): 141-144
4. Avşar, C., and Berber, İ., 2014. Plasmid profiling and antibiotics resisitance of Escherichia coli strains isolated from Mytilus galloprovincialis and seawater. Journal of Coastal Life Medicine 2(9): 689-693
5. Balta, F., Sandalli, C., Kayis, S., and Ozgumus, O. B., 2010. Molecular analysis of antimicrobial resistance in Yersinia ruckeri strains isolated from rainbow trout (Oncorhynchus mykiss) grown in commercial fish farms in Turkey. Bulletin of The European Association of Fish Pathologists. 30, 211-219
6. Baltas, H., Dalgic, G., Bayrak, E. Y., Sirin, M., Cevik, U., and Apaydin, G., 2016. Experimental study on copper uptake capacity in the Mediterranean mussel (Mytilus galloprovincialis). Environmental Science and Pollution Research, 23(11): 10983-10989
7. Bat, L., and Öztekin, H. C. 2016. Heavy Metals in Mytilus galloprovincialis, Rapana venosa and Eriphia verrucosa from the Black Sea Coasts of Turkey as Bioindicators of Pollution. Walailak Journal of Science and Technology, 13(9): 715-728
8. Brenner, D. J., and Farmer, J. J., 2015. Enterobacteriaceae. In Bergey's Manual of Systematics of Archaea and Bacteria: John Wiley & Sons, Ltd.

9. Capkin, E., Terzi, E., and Altinok, I., 2015. Occurrence of antibiotic resistance genes in culturable bacteria isolated from Turkish trout farms and their local aquatic environment. Diseases of Aquatic Organisms, 114(2): 127-137
10. CLSI, 2014. Clinical and laboratory standards institute. Performance standards for antimicrobial susceptibility testing, 17
11. Grevskott, D. H., Svanevik, C. S., Sunde, M., Wester, A. L., and Lunestad, B. T., 2017. Marine Bivalve Mollusks As Possible Indicators of Multidrug-Resistant Escherichia coli and Other Species of the Enterobacteriaceae Family. Frontiers in Microbiology, 8(24), doi:10.3389/fmicb.2017.00024
12. Guardabassi, L., Dijkshoorn, L., Collard, J. M., Olsen, J. E., and Dalsgaard, A., 2000. Distribution and in-vitro transfer of tetracycline resistance determinants in clinical and aquatic Acinetobacter strains. Journal of Medical Microbiology, 49(10): 929-936
13. Kacar, A., 2011. Some microbial characteristics of mussels (Mytilus galloprovincialis) in coastal city area. Environmental Science and Pollution Research, 18(8): 1384-1389
14. Ng, L. K., Martin, I., Alfa, M., and Mulvey, M., 2001. Multiplex PCR for the detection of tetracycline resistant genes. Molecular and Cellular Probes, 15(4): 209-215 Rees, E. E., Davidson, J., Fairbrother, J. M., St Hilaire, S., Saab, M., and McClure, J. T., 2015. Occurrence and antimicrobial resistance of Escherichia coli in oysters and mussels from Atlantic Canada. Foodborne Pathogens and Disease, 12(2): 164-169
15. Ryu, S.-H., Park, S.-G., Choi, S.-M., Hwang, Y.-O., Ham, H.-J., Kim, S.-U., et al., 2012. Antimicrobial resistance and resistance genes in Escherichia coli strains isolated from commercial fish and seafood. International Journal of Food Microbiology, 152(1-2): 14-18
16. Sandalli, C., Özgümüş, O. B., and Sevim, A., 2010. Characterization of tetracycline resistance genes in tetracycline-resistant Enterobacteriaceae obtained from a coliform collection. World Journal of Microbiology and Biotechnology, 26(11): 2099-2103