Balık Kökenli Mezofil Aeromonas ve Enterobacteriaceae İzolatlarının Tür Düzeyleri ve Genişlemiş Spektrumlu Beta Laktamaz ve Kolistin Direnç Profillerinin Belirlenmesi

Abstract

Çalışmada, kültür balıklarında Aeromonas ve Enterobacteriaceae'lerin tür düzeyleri ile antibiyotik direnç profillerinin belirlenmesi amaçlandı. Bu kapsamda toplam 100 Aeromonas ve Enterobacteriaceae izolatları 37 kültür balıklarından [levrek (n=14), çipura (n=14) ve gökkuşağı alabalık (n=9)] klasik kültür tekniği ile izole edildi ve MALDI- TOF MS sistemi ile tür düzeyleri belirlendi. Antibiyogramları CLSI guidelines (2021) ve EUROCAST (2023)'e göre yapıldı. İzolatlarda ayrıca Genişlemiş spektrumlu beta- laktamaz (GSBL) aktiviteleri çift disk sinerji testi ve blaTEM, blaSHV ve blaCTX-M ile kolistin direnç genleri (mcr-1, mcr-2, mcr-3, mcr-4 ve mcr-5) PZR ile belirlendi. İzole edilen 100 izolattan 8 tür Aeromonas [n=79; A. molluscorum (n=1), A.caviae (n=1), A.hydrophila (n=4), A. media (n=6), A.veronii (n=7), A. salmonicida (n=7), A. bestriarum (n=24) ve A. eucrenophila (n=25), 4 Aeromonas spp.] ve 11 tür Enterobacteriaceae [n=21; Providencia rustigianii (n=1), Lelliottia amnigena (n=1), Serratia (S.) liquefaciens (n=1), S. fronticola (n=1), Escherichia coli (n=1), Buttiauxella gaviniae (n=1), Morgonella morganii (n=1), Citrobacter (C.) brakii (n=1), C. gillenii (n=4), Moellerella wisconsensis (n=2) ve Hafnia alvei (n=7) ] identifiye edildi. İzolatlar pensilin [AMP (%75), AMC (%16) ve SAM (%75)]; sefolosporin [FEF (%3), CTX (%2), CRO (%6), CAZ (%4) ve CFX (%13)]; karbapenem (IMP (%11), ETP (%10), MEM (%3)]; monobaktam (ATM (%5)]; florokinolon [CIP (%7), LEV (%10)]; aminoglokozit [CN (%11) ve TOB (%56)], tetrasiklin [TE (%18)], glisilsiklin [TGC (%10)]; kinolon (NAL (%23)]; amfenikol [(C (%6)]; fofomisin [FF (%4)]; sülfonomid [(SXT (%5)] grubu antibiyotiklere karşı dirençli idi. Çoklu antibiyotik direnç gösteren izolat sayısı 46 (%46,n=100) idi. Bu izolatlardan 12'si Enterobacteriaceae familyasına [7 H. alvei, 2 C. gillenii, 1 C. brakii, 1 Buttiauxella gaviniae, 1 Morgonella morganii] dahil iken, 34 izolat ise Aeromonas türlerine (10 A. eucronophila, 9 A. bestriarum, 5 A. veronii, 4 A. salmonocida, 1 A. media, 1 A. molluscorum, 1 A. hydrophila, 3 Aeromonas spp.) ait idi. Üç GSBL geninden en az biri 60 (%60) izolatta belirlendi. En yaygın GSBL geni blaTEM (%55) iken bu geni blaSHV (%11) izledi. blaCTX-M ise sadece %6 idi. En az iki geni birlikte içeren izolat sayısı ise %11 idi. Her üç genden en az birini ihtiva eden Aeromonas izolat sayısı 46 (%58,23, n=79) iken en yaygın gen blaTEM (n=42), Enterobacteriaceae izolatlarında (n=19) bu üç genden en az birini ihtiva eden izolat sayısı 15 (%71,43, n=21) idi. Kolistin direnç genleri hiçbir izolatta saptanamadı. Sanger dizi analizi ile tür tayininde yetersiz kaldığı sonucuna varıldı.The aim of the study was to determine the species levels and antibiotic resistance profiles of Aeromonas and Enterobacteriaceae in cultured fish. In this context, a total of 100 Aeromonas and Enterobacteriaceae isolates were isolated from 37 cultured fish [sea bass (n=14), sea bream (n=14) and rainbow trout (n=9)] by classical culture technique and their species levels were determined by MALDI-TOF MS. Antibiograms were performed according to CLSI guidelines and EUROCAST. Extended spectrum beta-lactamase (ESBL) activities were determined by double disc synergy test and blaTEM, blaSHV and blaCTX-M and colistin resistance genes (mcr-1, mcr 2, mcr-3, mcr-4 and mcr-5) were determined by PCR. In addition, sequence test applied. As a results, isolates, 8 Aeromonas species [n=79; A. molluscorum (n=1), A. caviae (n=1), A. hydrophila (n=4), A. media (n=6), A. veronii (n=7), A. salmonicida (n=7), A. bestriarum (n=24), A. eucrenophila (n=25), 4 Aeromonas spp.] and 11 Enterobacteriaceae species [n=21; Providencia rustigianii (n=1), Lelliottia amnigena (n=1), Serratia (S. ) liquefaciens (n=1), S. fronticola (n=1), Escherichia coli (n=1), Buttiauxella gaviniae (n=1), Morgonella morganii (n=1), Citrobacter (C.) brakii (n=1), C. gillenii (n=4), Moellerella wisconsensis (n=2), Hafnia alvei (n=7) ] were identified. The isolates were treated with pencillin [AMP (75%), AMC (16%) and SAM (75%)]; cefolosporin [FEF (3%), CTX (2%), CRO (6%), CAZ (4%) and CFX (13%)]; carbapenem (IMP (11%), ETP (10%), MEM (3%)]; monobactam (ATM (5%)]; fluoroquinolone [CIP (7%), LEV (10%)]; aminoglycoside [CN (11%) and TOB (56%)], tetracycline [TE (18%)], glycylcycline [TGC (10%)]; quinolone [NAL (23%)]; amphenicol [(C (6%)]; fofomycin [FF (4%)]; sulphonomide [(SXT (5%)] group antibiotics. The MDR isolates was 46 (46%). While 12 isolates belonged to Enterobacteriaceae family [7 H. alvei, 2 C. gillenii, 1 C. brakii, 1 Buttiauxella gaviniae, 1 Morgonella morganii], 34 isolates belonged to Aeromonas species (10 A. eucronophila, 9 A. bestriarum, 5 A. veronii, 4 A. salmonocida, 1 A. media, 1 A. molluscorum, 1 A. hydrophila, 3 Aeromonas spp.). At leat one ESBL genes was identified in 60 (60%) isolates. The most common ESBL gene was blaTEM (55%), followed by blaSHV (11%). blaCTX-M was only 6%. The number of isolates containing at least two genes together was 11%. The number of Aeromonas isolates containing at least one of the three genes was 46 (58.23%, n=79), the most common gene was blaTEM (n=42), and the number of Enterobacteriaceae isolates (n=19) containing at least one of the three genes was 15 (71.43%, n=21). Colistin resistance genes were not detected. Sanger sequence analysis was insufficient for species determination.