Mezbaha Atık Sularından İzole Edilen Enterococcus Türlerinin Moleküler Karakterizasyonu

Yıl 2024, Cilt: 21 Sayı: 1, 55 - 64, 01.04.2024

Nuri Güngör , Dursun Alp Gündoğ , Candan Güngör , Nurhan Ertaş Onmaz

https://doi.org/10.32707/ercivet.1455401

Öz

Bu çalışmada, sığır mezbahası atık suyunun (MAS) önemli bir halk sağlığı riski yaratan antibiyotik dirençli Entero-coccus faecalis ve Enterococcus faecium türlerinin çevreye salınımında olası bir kaynak olarak rolünün araştırılması amaçlandı. Bu amaçla, büyükbaş hayvan mezbahalarından temin edilen 106 adet MAS örneğinden konvansiyonel metot ile izole edilen Enterococcus spp. suşları kantitatif gerçek zamanlı polimeraz zincir reaksiyonu (Q-PCR) ile konfir-me edildi ve konvansiyonel PCR ile karakterize edildi. İzolatların antimikrobiyal duyarlılıkları disk difüzyon yöntemi ile belirlendi. Çalışmada MAS örneklerinden elde edilen 95 (%89.6) enterokok izolatının 33’ü (%34.7) E. faecalis ve 10’u (%10.5) ise E. faecium olarak identifiye edildi. E. faecalis izolatlarının Quinupristin-dalfopristine, rifampin, tetrasiklin ve eritromisine karşı direnç oranları sırasıyla %90, %88, %75 ve %63 olarak belirlenirken E. faecium izolatlarının ilgili antibiyotiklere direnç oranları ise sırasıyla %90, %100, %80 ve %80 olarak belirlendi. Fakat, linezolid (%90), vankomi-sin (≥%70) ve penisilin (>%90) E. faecalis ve E. faecium izolatlarına karşı yüksek etkinlik gösterdi. Ayrıca E. faecalis izolatlarının 26’sı (%79), E. faecium izolatlarının tamamı (%100) çoklu antibiyotik direnci (ÇAD) gösterdi ve bu izolatlar-dan sırası ile %84’ünün ve %100’ünün ÇAD indeksleri 0.2’den yüksek idi. Sonuç olarak, analiz edilen MAS’ların tıbbi açıdan önemli antimikrobiyallere karşı yüksek oranda çoklu direnç sergileyen E. faecium ve E. faecalis izolatları için bir rezervuar görevi görebileceği ve bu organizmaların çevreye ve insanlara kolonizasyonunda önemli rol alabileceği tespit edildi. Bu nedenle, zengin mikrobiyotaya sahip MAS’ların halk sağlığı ve çevre üzerine olumsuz etkilerin en aza indiril-mesi için uygun arıtma stratejileri uygulanmalıdır.

Anahtar Kelimeler

E. faecalis, E. faecium, halk sağlığı, mezbaha atık suyu

Molecular Characterization of Enterococcus Species Isolated from Slaughterhouse Wastewater

Öz

In this study, it was aimed to investigate the role of cattle slaughterhouse wastewater (SWW) as a possible source of the antibiotic-resistant Enterococcus faecalis and Enterococcus faecium strains that pose a significant public health risk. For this purpose, Enterococcus spp. strains isolated by conventional method from 106 SWW samples ob-tained from cattle slaughterhouses were confirmed by quantitative real-time polymerase chain reaction (Q-PCR) and characterized by conventional PCR. Their antimicrobial susceptibilities were determined with disc diffusion method. Out of the 95 (89.6%) enterococci isolates, 33 (34.7%) were identified as E. faecalis and 10 (10.5%) as E. faecium. While E. faecalis isolates were resistant to Quinupristin-dalfopristin, rifampin, tetracycline and erythromycin at the rates of 90%, 88%, 75% and 63%, respectively; the resistance rates of E. faecium isolates to the relevant antibiotics were determined as 90%, 100%, 80% and 80%, respectively. However, linezolid (90%), vancomycin (≥70%), and penicillin (>90%) showed high efficacy against E. faecalis and E. faecium isolates. Additionally, 26 (79%) of the E. faecalis isolates and all (100%) E. faecium isolates exhibited multiple drug resistance (MDR) and their multiple antibiotic resistance (MAR) indexes were greater than 0.2 for 84% and 100% of these isolates, respectively. In conclusion, the analyzed SWWs can serve as a reservoir for E. feacium and E. faecalis isolates that exhibit high resistance to medically significant antimicrobials, and they can play a critical role in colonization on the environment and humans. Therefore, appropriate treatment strategies should be applied to minimize the negative effects of SWWs with rich microbiota on public health and environment.

Anahtar Kelimeler

E. faecalis, E. faecium, public health, slaughterhouse wastewater

Kaynakça

• Adegoke AA, Madu CE, Reddy P, Stenström TA, Okoh AI. Prevalence of vancomycin resistant En-terococcus in wastewater treatment plants and their recipients for reuse using PCR and MALDI-ToF MS. Front Environ Sci 2022; 9: 797992.
• Adeniji OO, Sibanda T, Okoh AI. Molecular detection of antibiotic resistance and virulence gene determinants of Enterococcus species isolated from coastal water in the Eastern Cape Province, South Africa. Int J Environ Stud 2021; 78(2): 208-27.
• Alipour M, Hajiesmaili R, Talebjannat M, Yahyapour Y. Identification and antimicrobial resistance of Enterococcus spp. isolated from the river and coastal waters in northern Iran. Sci World J 2014: 287458.
• Carey SA, Goldstein RER, Gibbs SG, Claye E, He X, Sapkota AR. Occurrence of vancomycin-resistant and-susceptible Enterococcus spp. in reclaimed water used for spray irrigation. Environ Res 2016; 147: 350-5.
• Clinical and Laboratory Standards Institute (CLSI). M100-performance Standards for Antimicrobial Susceptibility Testing. Thirty Third Edition 2023.
• Dec M, Stȩpień-Pyśniak D, Gnat S, Fratini F.; Urban-Chmiel R, Cerri D, Winiarczyk S, Turchi B. Antibi-otic susceptibility and virulence genes in Entero-coccus isolates from wild mammals living in Tusca-ny, Italy. Microb Drug Resist 2020; 26: 505-19.
• Dutka-Malen S, Evers S, Courvalin P. Detection of glycopeptides resistance genotypes and identification to the species level of clinically relevant entero-cocci by PCR. J Clin Microbiol 1995; 33(5): 24-7.
• Foyle L, Burnett M, Creaser A, Hens R, Keough J, Madin L, Price R, Smith H, Stone S, Kinobe RT. Prevalence and distribution of antimicrobial resistance in effluent wastewater from animal slaughter facilities: A systematic review. Environ Pollut 2023; 318: 120848.
• Gagetti P, Bonofiglio L, García Gabarrot G, Kaufman S, Mollerach M, Vigliarolo L, von Specht M, Toresani I, Lopardo HA. Resistencia a los β-lactámicos en enterococos. Rev Argent Microbiol 2019; 51(2): 179-83.
• Gotkowska-Płachta, A. The prevalence of virulent and multidrug-resistant enterococci in river water and in treated and untreated municipal and hospital wastewater. Int J Environ Res Public Health 2021; 18(2): 563.
• Güngör C, Gündoğ D.A., Onmaz NE. (2023). Mezba-ha ortamından izole edilen Enterococcus faecalis izolatlarının biyofilm oluşturma kapasitesi ve biyofilm ile ilişkili virülans genlerin varlığı. Bozok Vet Sci 2023; 4(1): 12-7.
• Hancock LE, Gilmore MS. Pathogenicity of entero-cocci. Fischetti VA, Novick RP, Ferretti JJ, Portnoy DA, Rood JI, eds. In: Gram-Positive Pathogens. Washington DC: ASM Press, 2000; pp. 251-8.
• Hao W, Shan X, Li D, Schwarz S, Zhang SM, Li XS, Du XD. Analysis of a poxtA- and optrA-co-carrying conjugative multiresistance plasmid from Entero-coccus faecalis. J Antimicrob Chemother 2019; 74: 1771-5.
• Hembach N, Bierbaum G, Schreiber C, Schwartz T. Facultative pathogenic bacteria and antibiotic resistance genes in swine livestock manure and clinical wastewater: A molecular biology comparison. Environ Poll 2022; 313: 120128.
• Igbinosa IH, Raje OC. Characterization of Enterococ-cus species isolated from abattoir environment in Benin City, Nigeria. Ife J Sci 2019; 21(3): 81-95.
• Igbinosa EO, Beshiru A, Odjadjare EEO. Diversity, antimicrobial characterization and biofilm formation of enterococci isolated from aquaculture and slaughterhouse sources in Benin City, Nigeria. Ife J Sci 2021; 22: 51-63.
• Jannati E, Khademi F, Manouchehrifar M, Maleki D, Amirmozaffari N, Sadat Nikbin V, Arzanlou M. Anti-biotic resistance and virulence potentials of E. fae-calis and E. faecium in hospital wastewater: A case study in Ardabil, Iran. J Water and Health 2023; 21(9): 1277-90.
• Kim MH, Moon DC, Kim SJ, Mechesso AF, Song HJ, Kang HY, Choi JH, Yoon SS, Lim SK. Nationwide surveillance on antimicrobial resistance profiles of Enterococcus faecium and Enterococcus faecalis Isolated from healthy food animals in South Korea, 2010 to 2019. Microorganisms 2021; 9(5): 925.
• Krawczyk B, Wityk P, Gałęcka M, Michalik M. The many faces of Enterococcus spp. commensal, pro-biotic and opportunistic pathogen. Microorganisms 2021; 7(9): 1900.
• Kwit R, Zając M, Śmiałowska-Węglińska A, Skarżyńska M, Bomba A, Lalak A, Skrzypiec E, Wojdat D, Koza W, Mikos-Wojewoda E. Prevalence of Enterococcus spp. and the whole-genome char-acteristics of Enterococcus faecium and Enterococ-cus faecalis strains isolated from free-living birds in Poland. Pathogens 2023; 12(6): 836.
• Maleki D, Manouchehrifar M, Kheljan MN, Mossavi SH, Jannati E, Doghaheh HP, Teimourpour R, Khademi F, Arzanlou M. Vancomycin-resistant Enterococcus species: antimicrobial resistance and virulence genes profile. Gene Rep 2021; 25: 101338.
• Mir R, Salari S, Najimi M, Rashki A. Determination of frequency, multiple antibiotic resistance index and resistotype of Salmonella spp. in chicken meat collected from southeast of Iran. Vet Med Sci 2022; 8(1): 229-36.
• Montealegre MC, Roh JH, Rae M, Davlieva MG, Singh KV, Shamoo Y, Murray BE. Differential penicillin-binding protein 5 (PBP5) levels in the Entero-coccus faecium clades with different levels of ampi-cillin resistance. Antimicrob Agents Chemother 2017; 61(1): e02034-16.
• Mwikuma G, Kainga H, Kallu SA, Nakajima C, Suzuki Y, Hang’ombe BM. Determination of the prevalence and antimicrobial resistance of Enterococcus fae-calis and Enterococcus faecium associated with poultry in four districts in Zambia. Antibiotics 2023;12(4): 657.
• Ni J, Long X, Wang M, Ma J, Sun Y, Wang W, Yu M, Yang H, Pan D, Tang B. Transmission of linezolid-resistant Enterococcus isolates carrying optrA and poxtA genes in slaughterhouses. Front Sustain Food Syst 2023; 7: 1179078.
• Olawale SI, Busayo OOM, Olatunji OI, Mariam M, Olayinka OS. Plasmid profiles and antibiotic susceptibility patterns of bacteria isolated from abat-toirs wastewater within Ilorin, Kwara, Nigeria. Irani-an J Microbiol 2020; 12(6): 547.
• Paschoalini BR, Nuñez KVM, Maffei JT, Langoni H, Guimarães FF, Gebara C, Freitas NE, Santos VG, Fidelis CD, Kappes R, Gonçalves MC, Silva NCC. The emergence of antimicrobial resistance and virulence characteristics in Enterococcus species isolated from bovine milk. Antibiotics 2023; 12(8): 1243.
• Savin M, Bierbaum G, Kreyenschmidt J, Schmithausen RM, Sib E, Schmoger S, Kasbohrer A, Hammerl JA. Clinically relevant Escherichia coli isolates from process waters and wastewater of poultry and pig slaughterhouses in Germany. Mi-croorganisms 2021; 9(4): 698.
• Sib E, Lenz-Plet F, Barabasch V, Klanke U, Savin M, Hembach N, Bierbaum, G. Bacteria isolated from hospital, municipal and slaughterhouse wastewaters show characteristic, different resistance profiles. Sci Total Environ 2020; 746: 140894.
• Solaiman S, Patterson R, Davey K, Katz Y, Payne‐Sturges D, Sapkota AR, Micallef SA. Effects of season and water type on the distribution and antimicrobial resistance of Enterococcus faecalis and Ent. faecium from surface and reclaimed water. J Appl Microbiol 2022; 133(2): 477-87.
• US Environmental Protection Agency. Method 1611.1: Enterococci in water by TaqMan quantitative polymerase chain reaction (qPCR). EPA-820-R-15-008. US, 2015.
• Xuan H, Yao X, Pan R, Gao Y, Wei J, Shao D, Liu K, Li Z, Qiu Y, Ma Z, Li B, Xia L. Antimicrobial resistance in Enterococcus faecium and Enterococcus faecalis isolates of swine origin from eighteen provinces in China. J Vet Med Sci 2021; 83(12): 1952-8.
• Yoon S, Kim YB, Seo KW, Ha JS, Noh EB, Lee YJ. Characteristics of linezolid-resistant Enterococcus faecalis isolates from broiler breeder farms. Poult Sci 2020; 99: 6055-61.
• Zaheer R, Cook SR, Barbieri R, Goji N, Cameron A, Petkau A, McAllister TA. Surveillance of Enterococ-cus spp. reveals distinct species and antimicrobial resistance diversity across a One-Health continuum. Sci Reports 2020; 10 (1): 3937.