İnsuyu Mağarası’ndan İzole Edilen Bakterilerin Antibiyotik Dirençliliğinin Araştırılması

Yıl 2022, , 40 - 48, 09.06.2022

Nermin Sarıgül , Fatma Seçil Koç Çelik

https://doi.org/10.29048/makufebed.1056387

Cited By: 1

Öz

Ekstrem çevrelerde farklı metabolik özellikteki mikroorganizmaların keşfi için yapılan çalışmalar her geçen yıl artmaktadır. Bu çalışma kapsamında ekstrem çevre olarak İnsuyu mağarasından örneklemeler yapılmıştır. Elde edilen izolatlarda antibiyotik dirençliliği kültürel ve moleküler biyoloji yöntemleri ile araştırılmıştır. Çoklu direnç gösteren 10 izolat ve duyarlı 3 izolat seçilerek 16S rRNA dizi analizi ile tür tayini yapılmıştır. Çoklu direnç gösteren Pseudomonas spp. (2G-2), Pseudomonas jesseni (2J), Sphingopyxis fribergensis (4D), Microbacterium yannicii (4M), Flavobacterium chungangense (6B), Rhodococcus spp. (7A), Flavobacterium resistens (9D), Pseudomonas spp. (K-4F), Pseudomonas spp. (K-4G), Buttiauxella agrestis (K-15A) ve duyarlılık gösteren Flavobacterium chungangense (K-1E), Rhodococcus erythropolis (K-11G) ve Pseudomonas spp. (K-15G) suşlarında antibiyotik direnç genleri taranmıştır. Pseudomonas jesseni (2J) ve Pseudomonas spp. (K-4G)’ de aminoglikozit 2’’-O-nükleotiltransferaz (aadB), beta laktamaz (blaCTXM3, blaSHV, blaTEM), aminoglikozit direnç proteini (strA), rifampin ADP- ribosiltransferaz (arr2- int2a), vankomisin (vanC) olmak üzere 8 farklı gen bölgesi saptanmıştır.

Anahtar Kelimeler

Antibiyotik direnci, ekstrem çevreler, çoklu ilaç direnci

Investigation of Antibiotic Resistance of Bacteria Isolated from Insuyu Cave, Burdur-Turkey

Öz

Studies for the discovery of microorganisms with different metabolic properties in extreme environments are increasing every year. Within the scope of this study, samples were taken from, an extreme environment, İnsuyu cave. Antibiotic resistance in the obtained isolates was investigated by cultural and molecular biology methods. As a result of Minimum Inhibition Concentration (MIC) test, 64 cefpodoxim, 6 rifampicin and gentamycin, 18 vancomycin, 15 ampicillin, 44 clindamycin, 48 penicillin resistant strains were detected. Pseudomonas spp. (2G-2), Pseudomonas jesseni (2J), Sphingopyxis fribergensis (4D), Microbacterium yannicii (4M), Flavobacterium chungangense (6B), Rhodococcus spp. (7A), Flavobacterium resistens (9D), Pseudomonas spp. (K-4F), Pseudomonas spp. (K-4G), Buttiauxella agrestis (K-15A) were found to have multidrug resistant by cultural methods. These bacteria and susceptible strains, Flavobacterium chungangense (K-1E), Rhodococcus erythropolis (K-11G) and Pseudomonas spp. (K-15G), as negative controls were selected for the identification of antibiotic resistance genes by PCR based methods. Eight different gene regions, aminoglycoside 2''-O-nucleotyltransferase (aadB), beta lactamase (blaCTXM3, blaSHV, blaTEM), aminoglycoside resistance protein (strA), rifampin ADP- ribosyltransferase (arr2- int2a), vancomycin (vanC) have been detected in Pseudomonas jesseni (2J) and Pseudomonas spp. (K-4G).

Anahtar Kelimeler

Antibiotic resistance, extreme environment, multidrug resistance

Kaynakça

• Bhullar, K. (2011). Investigation of antibiotic resistance in isolated Lechuguilla. M.Sc. Thesis, McMaster University, Department of Biochemistry.
• Carattoli, A. (2002). Use of a Light Cycler gyrA mutation assay for identification of ciprofloxacin‐resistant Campylobacter coli. FEMS Microbiology Letters, 214: 87-93.
• Chandrasekaran, S., Lalithakumari, D. (1998). Plasmid-mediated rifampicin resistance in Pseudomonas fluorescens. Journal of Medical Microbiology, 47: 197-200.
• Cheeptam, N., Sadoway, T., Rule, D., Watson, K., Moote, P., Soliman, L. (2013). Cure from the cave: Volcanic cave actinomycetes and their potential in drug discovery. International Journal of Speleology, 35-47.
• Davies, J., Davies, D. (2010). Origins and evolution of antibiotic resistance. Microbiology and Molecular Biology Reviews, 74: 417-433.
• Fonseca, E.L., Freitas F., Amorim, J.C., Vicente, C.P. (2008). Detection of New arr-4 and arr-5 Gene Cassettes in Clinical Pseudomonas aeruginosa and Klebsiella pneumoniae Strains from Brazil. Antimicrobial Agents and Chemotherapy, 52: 1865–1867.
• Houang, E., Chu, Y., Lo, W., Chu, K., Cheng, A. (2003). Epidemiology of Rifampin ADP-Ribosyltransferase (arr-2) and Metallo- -Lactamase (blaIMP-4) Gene Cassettes in Class 1 Integrons in Acinetobacter Strains Isolated from Blood Cultures in 1997 to 2000. Antimicrobial Agents and Chemotherapy, 47: 1382–1390.
• Lee, S. H.,]eoni, H., Lee, K.J. (2011). Evolution of TEM Beta-Iactamase genes identified by peR with newly designed primers in Korean clinical isolates. Clinical Microbiology and Infection, 7: 88-102.
• Levy, S. (2002). Factors impacting on the problem of antibiotic resistance. Journal of Antimicrobial Chemotherapy, 49: 25-30.
• Miele, A., Bandera, M., Goldstein, B.P. (1995). Use of Primers selective for vancomycin resistance genes to determine van Genotype in Enterococci and to study gene organization in VanA isolates. Antimicrobial Agents and Chemotherapy, 39: 1772–1778.
• Mir, R. A., Weppelmann, T.A., Johnson, J.A., Archer, D., Morris, J. G., Jeong, C.K. (2016). Identification and characterization of Cefotaxime resistant bacteria in beef cattle. Plos One, 11(9): e0163279; DOI: 10.1371/journal.pone.0163279
• Rajput, Y., Biswas, J., Rai, V. (2012). Potentiality test in antimicrobial activity and antibiotic sensitivity of subterranean Streptomyces strains isolated from 70 Kotumsar cave of India. The International Journal of Biological and Chemical Sciences, 6: 53-60.
• Sharma, J., Sharma, M., Ray, P. (2010). Detection of TEM & SHV genes in Escherichia coli & Klebsiella pneumoniae isolates in a tertiary care hospital from India. Indian Journal of Medical Research, 132: 332-336.
• Stankovic, N., Radulovic, V., Petkovic, M., Vuckovic, I., Jadranin, M., Vasiljevic, B. (2012). Streptomyces sp. JS520 produces exceptionally high quantities of undecylprodigiosin with antibacterial, antioxidative, and UV-protective properties. Applied Microbiology and Biotechnolohy, 96: 1217-31.
• Teixeira, B., Rodulfo, H., Carreno N., Guzman, M., Salazar, E., De Donato, M. (2016). Aminoglycoside Resistance Genes in Pseudomonas aeruginosa Isolates from Cumana, Venezuela. The Revista do Instituto de Medicina Tropical de São Paulo, 58, 13; DOI: 10.1590/S1678-9946201658013
• White, M. (2013). Living in a bacterial World: Antibiotic-resistant bacteria everywhere Pacific standard. http://www.psmag.com/science/living-bacterial-world-66260/
• Yücel, S., Yamaç, M. (2010). Selection of Streptomyces isolates from Turkish karstic caves against antibiotic resistant microorganisms. Journal of Pharmaceutical Sciences and Pharmacology, 23(1):1-6.