Antibiotic Resistance Profiles of Gram Negative Bacteria Isolated From Wastewater

Year 2024, Volume: 27 Issue: 3, 520 - 527

Merve Cora İnci Durukan Gülşen Uluçam Atay Ali Osman Kılıç

https://doi.org/10.18016/ksutarimdoga.vi.1248203

Abstract

Aims: The use of antibiotics, which have been used in the treatment of infections, has been increasing. Therefore, the occurrence of antibiotic-resistant bacteria and resistance genes, both in infections and in the environment, is also increasing. In settlements near the water, wastewater is commonly collected in certain centers and discharged into nearby waters after treatment. As a result, the waters are polluted with pathogenic and resistant microorganisms, posing a great danger to human health. In this study, it was aimed at determining the antibiotic resistance profiles of Gram-negative bacteria isolated from wastewater samples taken from Trabzon City Center. Material and methods: Samples taken from wastewater were inoculated on tryptic soy agar and eosin methylene blue media to analyze the growing microorganisms. Thirty-six Gram-negative bacteria were included in the study. Resistance in bacteria was investigated by the disk diffusion method. Microorganisms with resistance were identified with the MALDI-TOF MS and BD Phoenix automated microbiology systems, and resistance profiles were obtained with the automated systems. The presence of blaSHV, blaTEM, blaOXA, blaCTX-M, integrase 1, integrase 2, and integron in resistant bacteria was investigated by the PCR method, and the transmission mechanisms of these genes were examined by transformation and conjugation experiments. In addition, the production of Extended Spectrum Beta Lactamase (ESBL) in isolates resistant to antibiotics was investigated by a double-disc synergy test.
Results: As a result, in the current study, antibiotic resistance was found in 14 of 36 isolates. The presence of ESBL and blaoxa was shown to be isolated. It was determined that two isolates had plasmids. It was also shown that ampicillin resistance in one isolate was transformed with a conjugative plasmid, and kanamycin resistance was transformed with a non-conjugative plasmid. Conclusions: The detection of the presence of plasmids in some isolates carrying resistance suggests that antibiotic resistance may spread among bacteria in wastewater and may have adverse effects on living things.

Keywords

Antibiotic resistance, PCR, Bacterial transformation

Atık Sulardan İzole Edilen Gram Negatif Bakterilerin Antibiyotik Direnç Profillerinin Belirlenmesi

Abstract

İnsan ve hayvanlardaki enfeksiyonların tedavisinde ve endüstride çeşitli amaçlarla yer alan antibiyotiklerin kullanımının artması çevresel ortamlarda antibiyotiğe dirençli bakterilerin ve direnç genlerinin daha sık tespit edilmesine neden olmaktadır. Su kenarındaki yerleşim yerlerinde atık sular arıtma işleminden sonra yakındaki sulara boşaltılmaktadır. Bunun sonucunda, sular patojen ve dirençli mikroorganizmalarla kirlenerek insan sağlığı için büyük bir tehlike oluşturmaktadır. Bu çalışmada, Trabzon şehir merkezinden alınan atık su örneklerinden Gram negatif bakteriler izole edilerek antibiyotik direnç profillerinin belirlenmesi amaçlanmıştır. Atık sulardan alınan örnekler triptik soy agar ve eozin metilen mavisi besiyerlerine ekim yapılarak üreyen mikroorganizmalar analiz edilmiştir. Çalışmaya Gram negatif 36 bakteri izolatı dahil edilmiştir. Bakterilerdeki direnç disk difüzyon yöntemi ile araştırılmıştır. Antimikrobiyal direnç saptanan mikroorganizmalar MALDI-TOF MS ve BD Phoenix otomatize mikrobiyoloji sistemi ile tanımlanarak otomatize sistem ile direnç profilleri çıkarılmıştır. Dirençli bakterilerde blaSHV, blaTEM, blaOXA, blaCTX-M, integraz 1, integraz 2 ve integron varlığı PZR yöntemi ile araştırılarak bu genlerin aktarım mekanizmaları transformasyon ve konjugasyon deneyleri ile incelenmiştir. Ayrıca, antibiyotiklere dirençli izolatlarda Genişlemiş Spektrumlu Beta Laktamaz (GSBL) üretimi çift disk sinerji testi ile araştırılmıştır. Sonuç olarak bu çalışmada 14 izolatta antibiyotik direnci olduğu, bir izolatta GSBL ve blaoxa varlığı tespit edilmiştir. İki izolatın plazmit taşıdığı ve bir izolatın ampisilin direncinin konjugatif plazmitle, kanamisin direncinin ise konjugatif olmayan bir plazmitle transforme olduğu belirlenmiştir. Direnç taşıyan bazı izolatlarda plazmit varlığının tespit edilmesi; antibiyotik direncinin atık sularda bulunan bakteriler arasında yayılabileceğini ve canlılar üzerinde olumsuz etkiler olabileceğini düşündürmektedir.

Keywords

Bakteriyel transformasyon, Antibiyotik direnci, PZR

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