Nevşehir İlindeki Kesimhanelerden İzole Edilen Staphylococcus aureus İzolatlarının Moleküler Tiplendirilmesi ve Vankomisin Dirençliliğinin İncelenmesi

Year 2020, , 227 - 234, 01.12.2020

Ömer Tolga Yılmaz Harun Hızlısoy

https://doi.org/10.32707/ercivet.828369

Abstract

Bu çalışma, kesimhanelerde Staphylococcus aureus’un mevcudiyetini, izolatların moleküler tiplendirmesini ve izolatların vankomisin antibiyotiğine duyarlılığını tespit etmeyi amaçlamaktadır. Nevşehir ilindeki üç farklı kesimhane-den sıvap ile alınan; sığır karkası, duvar, bıçak, kesme tahtası yüzeyleri ve kesimhane atık su örnekleri çalışma kapsa-mında materyal olarak kullanılmıştır. Her bir kesimhaneden; 10 adet sığır karkası yüzeyi, 10 adet duvar yüzeyi, 10 adet bıçak yüzeyi, 10 adet kesme tahtası yüzeyi ve 10 adet atık su olmak üzere, bir kesimhaneden toplam 50 adet; üç ke-simhaneden toplam 150 adet numune incelenmiştir. Etken izolasyonunu takiben izolatların identifikasyonu fenotipik ve moleküler yöntemlerle (PCR) yapılmıştır. İzolatların vankomisine duyarlılıkları E test ile test edilmiştir. Ayrıca izolatların genetik yakınlıkları Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) ile araştırılmıştır. Çalışmada 150 örnekten 150 izolat elde edilmiş, bu izolatların 65’i (%43.3) fenotipik olarak koagülaz pozitif stafilokok olarak tespit edilmiştir. Bu 65 izolatın 6’sı (%4) ise PCR sonucu S. aureus olarak identifiye edilmiştir. Elde edilen S. aureus izolatları-nın tümünün (duyarlı: ≤2 μg/ml) vankomisine duyarlı olduğu saptanmıştır. ERIC-PCR analizi sonucu izolatların genoti-pik olarak farklı olduğu saptanmıştır. Hem gıda toksikasyonlarının önüne geçmek hem de karkas kalitesini artırmak ve kırmızı etin raf ömrünü uzatmak için, kesimhanede hijyen koşullarına dikkat edilmelidir. Yapılan antibiyotik duyarlılık test sonucu, direnç tespit edilmemesi olumlu olmakla birlikte, antibiyotiklerin bilinçsiz kullanımından kaçınılmalı ve kul-lanımları kontrol altında tutulmalıdır.

Keywords

E test, halk sağlığı, kesimhane, vankomisin, VRSA

Molecular Typing of Staphylococcus aureus Isolates Obtained from Slaughterhouses in Nevsehir Province (Turkey) and Investigation of Vancomycin Resistance of The Isolates

Abstract

This study aims to determine the presence of Staphylococcus aureus in the slaughter houses, the molecu-lar typing of the isolates and the susceptibility of the isolates to the vancomycin antibiotic. The samples collected from cattle carcass, wall, knife, wastewater and cutting board from three different slaughterhouses in the Province of Nevşehir, in the scope of the study, were used as material. Fifty swab samples of 10 beef carcass surfaces, 10 wall surface, 10 blade surface, 10 cutting board surface and 10 waste water were taken from each slaughterhouse; a total of 150 swab samples were analyzed. The identification of the isolates was performed by phenotypic and molecular methods (PCR) following the isolation of the agents. The susceptibility of the isolates to vancomycin was tested by E test. In addition, genetic proximity of the isolates was investigated by Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR). In this study, 150 isolates were obtained from 150 samples. 65 (43.3%) of these isolates were iden-tified as coagulase positive staphylococci and 6 (4%) of 150 isolates were identified as S. aureus. All S. aureus isolates (susceptible: ≤2 μg / ml; moderately sensitive: 4-8 μg / ml; resistant: ≥16 μg / ml) were found to be susceptible to van-comycin. According to the ERIC-PCR analysis, isolates have been determined to be genotypically different. Special attention should be paid to hygiene conditions in slaughterhouses, both to prevent food poisoning and to improve car-cass quality, and to extend the shelf life of red meat. Although it is pleasing that no resistance is detected as a result of our antimicrobial susceptibility test; unconscious use of antibiotics should be avoided, and their use should be kept under control.

Keywords

E Test, public health, slaughterhouse, vancomycine

References

• Adugna F, Pal M, Girmay G. Prevalence and antibi-ogram assessment of Staphylococcus aureus in beef at municipal abattoir and but chers hops in Addis Ababa, Ethiopia. Biomed Res Int 2018: 1-7.
• Abdollahi S, Ramazanzadeh R, Kalantar E, Zamani S. Molecular epidemiology of Staphylococcus au-reus with ERIC-PCR method. Bull Env Pharmacol Life Sci 2014; 3(3): 158-65.
• Adwan G, Shaheen H, Adwan K, Barakat A. Molecu-lar characterization of methicillin resistant Staphy-lococcus aureus isolated from hospitals environ-ments and patients in Northern Palestine. Epide-miol Biostat Public Health 2015;12(3): e11183.
• Arslan E, Mutlu EG. Genotyping of Staphylococcus aureus strains isolated from bovine mastitis in Tur-key by using ERIC-PCR method. Pakistan J Zool 2016; 48(6): 1747-52.
• Aydin A, Muratoglu K, Sudagidan M, Bostan K, Okuk-lu B, Harsa S. Prevalence and antibiotic resistance of foodborne Staphylococcus aureus isolates in Turkey. Foodborne Pathog Dis 2011; 8(1): 63-9.
• Barber M. Methicillin-resistant Staphylococci. J Clin Path 1961; 14: 385-92.
• Bersisa A, Tulu D, Negera, C. Investigation of bacte-riological quality of meat from abattoir and butcher shops in Bishoftu, Central Ethiopia. Int J Food Microbiol 2019: 1-8.
• Beyene T, Hayishe H, Gizaw F, Beyi AF, Abunna F, Mammo B, Ayana D, Walktole H, Abdi RD. Preva-lence and antimicrobial resistance profile of Staphylococcus in dairy farms, abattoir and hu-mans in Addis Ababa, Ethiopia. BMC Research Notes 2017; 10(1): 171.
• Birhanu W, Weldegebriel S, Bassazin G, Mitku F, Birku L, Tadesse M, Assesment of microbiological quality and meat handling practices in butcher shops and abattoir found in Gondar town, Ethiopia. Int J Microbiol Res 2017; 8(2): 59-68.
• Crago B, Ferrato, C, Drews SJ, Svenson LW, Tyrrell G, Louie M. Prevalence of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) in food samples associated with food borne illness in Al-berta, Canada from 2007. Food Microbiol 2010; 32(1): 202-5.
• Cha JO, Lee JK, Jung YH, Yoo JI, Park YK, Kim BS, Lee YS. Molecular analysis of Staphylococcus aureus isolates associated with staphylococcal food poisoning in South Korea. J Appl Microbiol 2006; 101(4): 864-71.
• CLSI Performance Standards for Antimicrobial Sus-ceptibility Testing. Twenty Seventh Edition. (CLSI supplement M100 Clinicaland Laboratory Stan-dards Institute), Wayne. Pennsylvania, USA, 2017: 56-63.
• Desmarchelier PM, Higgs GM, Mills L, Sullivan AM,Vanderlinde PB. Incidence of coagulase positive Staphylococcus on beef carcasses in three Austra-lia abattoirs. Int J Food Microbiol 1999; 47: 222-9.
• Dongyou L. Molecular Detection of Foodborne Patho-gens. First Edition. Boca Raton: CRC Press, 2010; p. 245-55.
• Ertaş Onmaz N, Datlı S. Bir büyükbaş hayvan kesim-hanesinde Escherichia coli O157:H7 varlığının IMS-PZR teknikleri ile araştırılması. Eurasian J Vet Sci 2018; 34(1): 49-55.
• Fijalkowski K, Peitler D, Karakulska J. Staphylococci isolated from ready-to-eat meat identification, anti-biotic resistance and toxin gene profile. Int J Food Microbiol 2016; 238: 113-20.
• Gardete S, Tomasz A. Mechanisms of vancomycin resistance in Staphylococcus aureus. J Clin Invest 2014;124 (7): 2836-40.
• Gomez-Gil B, Roque A, Turnbull JF. The use and selection of probiotic bacteria for use in the culture of larval aquatic organisms. Aquaculture 2000; 191: 259-70.
• Gowda Tanuja KGM, Latha C, Sunil B, Van Damme I. Occurrence and antibiotic susceptibility of Liste-ria species and Staphylococcus aureus in cattle slaughterhouses of Kerala, South India. Foodbor-ne Pathog Dis 2017; 14(10): 573-9.
• Hızlısoy H, Ertaş Onmaz N, Karadal F, Al S, Yıldırım Y, Gönülalan Z, Kılıc H. Antibiotic resistance gene profiles of Staphylococcus aureus isolated from foods of animal origin. Kafkas Univ Vet Fak Derg 2018; 24(2): 244-9.
• Howden BP, Davies JK, Johnson PD, Stinear TP, Grayson ML. Reduced vancomycin susceptibility in Staphylococcus aureus, including vancomycin intermediate and heterogeneous vancomycin-intermediate strains: resistance mechanisms, labo-ratory detection, and clinical implications. Clin Mic-robiol Rev 2010; 23(1): 99-139.
• Keyvan E ve Özdemir H. Sığır karkaslarında Staphy-lococcus aureus’un varlığı, enterotoksijenik özellik-leri ve antimikrobiyal dirençliliği. Ankara Üniv Vet Fak Derg 2016; 63(1): 17-23.
• Keyvan E. Sığır Karkaslarında Staphylococcus au-reus’un varlığı, karakterizasyonu ve antimikrobiyal dirençliliğinin belirlenmesi. Doktora tezi, Ankara Üniv Sağ Bil Ens, Ankara, 2014; s. 11-7.
• Özdemir H, Şireli UT, Can HY. Determination of mic-robial surface contamination on beef carcasses. Arch Lebensmittelhyg 2010; 61(1): 27-30.
• Sancak B. Staphylococcus aureus ve antibiyotik direnci. Mikrobiyol Bul 2011; 45(3): 568-71.
• Sorrell TC, Packham DR, Shanker S, Foldes M, Mun-ro R. Vancomycin therapy for methicillin-resistant Staphylococcus aureus. Ann Intern Med 1982; 97(3): 344-50.
• Şahal M. Süt ve besi hayvancılığında antibiyotik kul-lanımı. Bilinçli Antibiyotik Kullanımı ve Antimikrobi-yel Direnç Sempozyumu. 18 Ekim, 2012; Gıda Tarım ve Hayvancılık Bakanlığı, Ankara-Türkiye.
• Tanih NF, Sekwadi E, Ndip RN, Bessong PO. Detec-tion of pathogenic Escherichia coli and Staphylo-coccus aureus from cattle and pigs slaughtered in abattoirs in Vhembe District, South Africa. Sci World J 2015: 1-8.
• Terzi G, Gücükoğlu A, Çadırcı Ö, Uyanik T, Alişarlı M. Serotyping and antibiotic susceptibility of Liste-ria monocytogenes isolated from ready-to-eatfoods in Samsun, Turkey. Turk J Vet Anim Sci 2015; 39: 211-7.
• Tong, SY, Davis JS, Eichenberger E, Holland TL, Fowler VG Jr. Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifesta-tions, and management. Clin Microbiol Rev 2015; 28(3): 603-61.
• Veras, JF, do Carmo LS, Tong LC, Shupp JW, Cum-mings C, dos Santos DA, Cerqueira MMOP, Canti-ni A, Nicoli JR, Jett M. A study of the enterotoxige-nicity of coagulase-negative and coagulase-positive staphylococcal isolates from food poiso-ning outbreaks in Minas Gerai Brazil. Int J Infect Dis 2008; 12: 410-5.
• Ye Y, Jiang Q, Wu Q, Zhang J, Lu J, Lin L. The cha-racterization and comparison of Staphylococcus aureus by antibiotics usceptibility testing, entero-bacterial repetitive intergenic consensus-polymerase chain reaction, and random amplified polymorphic DNA-polymerase chain reaction. Fo-odborne Pathog Dis 2012; 9(2): 168-71.
• Yıldırım Y, Ertaş Onmaz N, Gonulalan Z, Al S, Yildi-rim A, Karadal F, Hızlısoy H, Pamuk Ş. Microbiolo-gicalquality of pastrami and associated surfaces at thepoint of sale in Kayseri Turkey. Public Health 2017; 146: 152-8.
• Yildirim T, Sadati F, Kocaman B, Siriken B. Staphylo-coccus aureus and Staphylococcal enterotoxin detection in raw milk and cheese origin coagulase positive isolates. Int J Sci Lett (IJSL) 2019; 1(1): 30-41.
• Yılmaz İ, Gümüş T. Sığır karkaslarının mikrobiyolojik kalitesinin belirlenmesi, Onuncu Gıda Kongresi. 21-23 Mayıs 2004; Atatürk Üniversitesi, Erzurum-Türkiye.