شناسایی مولکولی ژن‌های بتالاکتاماز (PER ،VEB و GES) درسالمونلا انتریتیدیس جدا شده از گوشت طیور با روش Multiplex-PCR
محورهای موضوعی :
علوم و صنایع غذایی
مهشید اسفندیان
1
(دانش آموخته کارشناسی ارشد گروه میکروبیولوژی، دانشکده علوم پایه، واحد ساوه، دانشگاه آزاد اسلامی، ساوه، ایران)
غلامعلی مرادلی
2
(استادیار گروه میکروبیولوژی، دانشکده علوم پایه، واحد ساوه، دانشگاه آزاد اسلامی، ساوه، ایران)
تاریخ دریافت : 1395/07/11
تاریخ پذیرش : 1397/03/04
تاریخ انتشار : 1397/07/01
کلید واژه:
سالمونلا انتریتیدیس,
blaVEB,
blaPER,
blaGES,
چکیده مقاله :
سالمونلوز یک بیماری مهم در انسان و گونه‌های حیوانات است که به‌وسیله سرووارهای مختلف سالمونلاانتریکا ایجاد می‌شود. سرووار انتریتیدیس یکی از شایع‌ترین سرووارها در انسان و طیور و هم‌چنین عامل بیماری منتقله از طریق غذا می‌باشد. هدف از این مطالعه بررسی حضور ژن‌های blaPER ،blaVEB و blaGES در سویه‌های سالمونلا انتریتیدیس جداسازی شده از گوشت مرغ می‌باشد. در این مطالعه، تعداد 60 نمونه سالمونلا جداسازی شده از گوشت مرغ جمع‌آوری و با استفاده از روش‌های کشت و بیوشیمیایی تأیید شد. سروتایپینگ نمونه‌ها با استفاده از آنتی سرمه‌ای O و H انجام شد. آزمون Multiplex-PCR جهت شناسایی ژن‌های مقاومت به آنتی‌بیوتیک‌های بتالاکتام وسیع الطیف شامل blaPER ،blaVEB و blaGES بر روی نمونه‌ها صورت گرفت. نتایج سروتایپینگ نشان داد هر 60 نمونه مربوط به گروه D و سرووار انتریتیدیس بودند. از 60 نمونه مورد بررسی 3 نمونه دارای ژن blaPER، 2 نمونه دارای ژن blaVEB و 1 نمونه دارای ژن blaGES بودند. نتایج آنتی بیوگرام نشان داد بیشترین مقاومت به آنتی‌بیوتیک سفتازیدیم (64%) و بیشترین حساسیت به آنتی‌بیوتیک‌های مروپنم و سفتی زوکسیم (92%) بود. با توجه به این‌که سالمونلا یک باکتری منتقله از راه غذا می‌باشد حضور ژن‌های بتالاکتاماز وسیع الطیف در نمونه‌های مواد غذایی هرچند به میزان کم می‌تواند هشداری جدی باشد.
چکیده انگلیسی:
Salmonellosis is an important disease in animals and human which is caused by different serovars of Salmonellaenterica. SerovarsEnteritidis is one of the most prevalent sorvars in animals and poultry and also is the food-borne pathogen. The aim of this study is to identify the prevalence of blaVEB, blaPER and blaGES in SalmonellaEnteritidis isolated from chicken samples.In this study, 60 Salmonella samples isolated from chickens were collected and confirmed as Salmonella by culture and biochemical tests. Serotyping was performed using O and H antisera. Multiplex-PCR was done for identification of ESBL genes including blaPER, blaVEB and blaGES. The serotyping results showed that all of the 60 samples belonged to group D and serovarEnteritidis. Among 60 samples, 3 had the blaPER, 2 had the blaVEb and 1harboured the blaGES gene.Antibiogram results showed the most resistance was to ceftazidime (64%) and the most susceptibility was to meropeneme and ceftizoxime (92%). As regards Salmonella is a food-borne bacteria, the presence of ESBL in food samples even in small numbers can be a serious alarm.
منابع و مأخذ:
· Ambler, R. P., Coulson, A. F., Frere, J. M., Ghuysen, J. M., Joris, B. Forsman, M., et al., (1991). A standard numbering scheme for the class A. beta-lactamases. Biochemistry Journal. 276: 269–270.
· Alikhani, M. Y., KarimiTabar, Z., Mihani, F., Kalantar, E., Karami, P., Sadeghi, M., et al., (2014). Antimicrobial resistance patterns and prevalence of blaPER-1 and blaVEB-1 genes among ESBL-producing Pseudomonas aeruginosaisolatesin West of Iran. Jundishapur Journal Microbiology. 7(1): e8888.
· Bauer, A., Kirby, W. M., Sherris, J. C. and Truck, M. (1996). Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology. 45: 493-496.
· Braden Christopher, R. (2006). Salmonellaenterica serotype Enteritidis and egg; a national apidemic in the United States.Food safety, 43: 512-17.
· Campos, J., Pichel, M., Vaz, T.M.I., Tavechio, A.T., Fernandes, S.A., Munoz, N., et al., (2012). Building PulseNet Latin American and Caribbean Salmonella regional database:first conclusions of genetic subtypes of S.Typhi, S. Typhimurium and S. Enteritidis circulating in six countries of the region. Food Research International. 45: 1030e1036.
· Castilla, K.S., de Gobbi, D.D., Moreno, L.Z., Paixao, R., Coutinho, T.A., dosSantos, J.L., et al., (2012). Characterization of Haemophilusparasuisisolated from Brazilian swine through serotyping, AFLP and PFGE. Res Veterinary Science. 92: 366e371.
· Clinical and Laboratory Standards Institute (CLSI) (2011): Performance Standards for Antimicrobial susceptibility Testing; twenty first Informational Supplement. M100-S21. Wayne, PA: CLSI.
· Cortez, A. L. L., Carvalho, A. C. F. B., Ikunob, A. A., Burger, K. P., Vidal-Martins, A. M. C. (2006). Identification of Salmonella spp. Isolates from chicken abattoirs by multiplex-PCR.Veterinary Science. 81: 340-344.
· Dallenne C., Da Costa A, Decré D., Favier C. Arlet G. (2010). Development of a set of multiplex PCR assays for the detection of genes encoding important β-lactamases in Enterobacteriaceae. Journal Antimicrobial Chemotherapy. 12: 498.
· Egli, T., Koster, W., Meile, L. (2002). Pathogenic microbes in water and food: changes and challenges. FEMS Microbiology Review. 26 (2): 111–12.
· Ezatpanah E, Moradi Bidhendi S, Khaki P, Ghaderi R, Seyedan Jasebi E, Moghtadaee Far S. (2013). Isolation, serotyping and antibiotic-resistance pattern of isolated Salmonella from chicken of Arak. Iranian Veterinary Journal. 9 (2): 88-96. [In Persian]
· Hidalgo, L., Hopkins, K. L., Wareham, D. W., Gutierrez B., Gonzalez-Zorn, B. (2012). Association of Extended-Spectrum -Lactamase VEB-5 and 16S rRNAmethyltransferasearmA in Salmonella entericafrom the United Kingdom. Antimicrobial Agents Chemotherapy. 56(9): 4985–4987.
· Poirel, L., Bonnin R. A., Nordmann, P. (2012). Genetic support and diversity of acquired extended-spectrum b-lactamases in Gram-negative rods. Infection Genetic Evoloution. 12:883–893.
· Ranjbar R, Sarshar M, Morovvati S. (2012). A Study of Ribotype Patterns of Salmonella Enterica Serovar Enteritidis Strains Isolated in Tehran, Iran. Journal Isfahan Medical School. 30(180): 238-247. [In Persian]
· Ranjbar R, Naghoni A. (2014). Class 1 integron-mediated antibiotic resistance in Salmonella enterica strains isolated in Tehran, Iran. Iranian Journal of Medical Microbiolology.7 (4):16-23. [In Persian]
· Rayamajhi, N., Kang, S. G., Kang, M. L., Lee, H. S., Park K. Y., Yoo, H. S. (2008). Assessment of antibiotic resistance phenotype and integrons in SalmonellaentericaserovarTyphimurium isolated from swine. Journal Veterinary Medical Science. 70(10):1133-7.
· Salyers, A. A. and Whitt, D. D. (2005). Revenge of the Microbes: How bacterial resistance is undermining the antibiotic miracle. ASM Press, Washington, DC.
· Soltan-Dallal M. M., SharifiYazdi, M. K., Mirzaei, N.,Kalantar., E. (2014). Prevalence of Salmonella spp. in packed and unpacked red meat and chicken in South of Tehran. Jundishapur Journal Microbiology. 7(4): 9254-58.
· Tajbakhsh, M., YaghoobiAvini, M., Alikhajeh, J., Tajeddin, E., Rahbar, M., Eslami, P., et al., (2016). Emergence of blaCTX-M-15, blaTEM-169 and blaPER-1 extended-spectrum β-lactamase genes among different Salmonella entericaserovars fromhuman faecal samples. Infection Disease. 48(7):550-6.
· Tajbakhsh M, Yaghoobi Avini M, Ali Khajeh J, Alebouyeh M, Nazemalhosseini Mojarad E, Zali MR. (2012). Increased-Resistance Phenotype Resulted from Elevated β-Lactamase Enzyme Activity in Salmonella Clinical Isolates. Journal Isfahan Medical School. 30(178): 1-11. [In Persian]
· Wang, Y., Yang, B., Wu, Y., Zhang, Z., Meng, X., Xi, M., et al., (2015). Molecular characterization of Salmonella enterica serovar enteritidis on retail raw poultry in six provinces and two National cities in China. Food Microbiology. 46: 74-80.
· Wu, H., Wang, Y., Wu, Y., Qiao, J., Li, H., Zheng, S., et al., (2015). Emergence of β-Lactamases and extended-spectrum β-Lactamases (ESBLs) producing Salmonellain retail raw chicken in China. Foodborne Pathology Disease. 12(3): 228-235.
· Wu, H., Xia, X., Cui, Y, Hu, Y., Xi, M., Wang, X., et al., (2013). Prevalence of extended-spectrum β-Lactamase–producing Salmonella on retail chicken in six provinces and two national cities in the people’s republic of China. Journal Food Protection. 76(12): 2040–2044.
Yahyavi Firozabadi A, Sedighi khavidak S, Soltan Dallal M.M. (2016). The Molecular epidemiology evaluation and antibiotic resistance patterns of Salmonella isolated in food outbreaks in Yazd province by culture and PCR methods. Razi Journal Medical Science. 23(144): 1-8. [In Persian]
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· Ambler, R. P., Coulson, A. F., Frere, J. M., Ghuysen, J. M., Joris, B. Forsman, M., et al., (1991). A standard numbering scheme for the class A. beta-lactamases. Biochemistry Journal. 276: 269–270.
· Alikhani, M. Y., KarimiTabar, Z., Mihani, F., Kalantar, E., Karami, P., Sadeghi, M., et al., (2014). Antimicrobial resistance patterns and prevalence of blaPER-1 and blaVEB-1 genes among ESBL-producing Pseudomonas aeruginosaisolatesin West of Iran. Jundishapur Journal Microbiology. 7(1): e8888.
· Bauer, A., Kirby, W. M., Sherris, J. C. and Truck, M. (1996). Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology. 45: 493-496.
· Braden Christopher, R. (2006). Salmonellaenterica serotype Enteritidis and egg; a national apidemic in the United States.Food safety, 43: 512-17.
· Campos, J., Pichel, M., Vaz, T.M.I., Tavechio, A.T., Fernandes, S.A., Munoz, N., et al., (2012). Building PulseNet Latin American and Caribbean Salmonella regional database:first conclusions of genetic subtypes of S.Typhi, S. Typhimurium and S. Enteritidis circulating in six countries of the region. Food Research International. 45: 1030e1036.
· Castilla, K.S., de Gobbi, D.D., Moreno, L.Z., Paixao, R., Coutinho, T.A., dosSantos, J.L., et al., (2012). Characterization of Haemophilusparasuisisolated from Brazilian swine through serotyping, AFLP and PFGE. Res Veterinary Science. 92: 366e371.
· Clinical and Laboratory Standards Institute (CLSI) (2011): Performance Standards for Antimicrobial susceptibility Testing; twenty first Informational Supplement. M100-S21. Wayne, PA: CLSI.
· Cortez, A. L. L., Carvalho, A. C. F. B., Ikunob, A. A., Burger, K. P., Vidal-Martins, A. M. C. (2006). Identification of Salmonella spp. Isolates from chicken abattoirs by multiplex-PCR.Veterinary Science. 81: 340-344.
· Dallenne C., Da Costa A, Decré D., Favier C. Arlet G. (2010). Development of a set of multiplex PCR assays for the detection of genes encoding important β-lactamases in Enterobacteriaceae. Journal Antimicrobial Chemotherapy. 12: 498.
· Egli, T., Koster, W., Meile, L. (2002). Pathogenic microbes in water and food: changes and challenges. FEMS Microbiology Review. 26 (2): 111–12.
· Ezatpanah E, Moradi Bidhendi S, Khaki P, Ghaderi R, Seyedan Jasebi E, Moghtadaee Far S. (2013). Isolation, serotyping and antibiotic-resistance pattern of isolated Salmonella from chicken of Arak. Iranian Veterinary Journal. 9 (2): 88-96. [In Persian]
· Hidalgo, L., Hopkins, K. L., Wareham, D. W., Gutierrez B., Gonzalez-Zorn, B. (2012). Association of Extended-Spectrum -Lactamase VEB-5 and 16S rRNAmethyltransferasearmA in Salmonella entericafrom the United Kingdom. Antimicrobial Agents Chemotherapy. 56(9): 4985–4987.
· Poirel, L., Bonnin R. A., Nordmann, P. (2012). Genetic support and diversity of acquired extended-spectrum b-lactamases in Gram-negative rods. Infection Genetic Evoloution. 12:883–893.
· Ranjbar R, Sarshar M, Morovvati S. (2012). A Study of Ribotype Patterns of Salmonella Enterica Serovar Enteritidis Strains Isolated in Tehran, Iran. Journal Isfahan Medical School. 30(180): 238-247. [In Persian]
· Ranjbar R, Naghoni A. (2014). Class 1 integron-mediated antibiotic resistance in Salmonella enterica strains isolated in Tehran, Iran. Iranian Journal of Medical Microbiolology.7 (4):16-23. [In Persian]
· Rayamajhi, N., Kang, S. G., Kang, M. L., Lee, H. S., Park K. Y., Yoo, H. S. (2008). Assessment of antibiotic resistance phenotype and integrons in SalmonellaentericaserovarTyphimurium isolated from swine. Journal Veterinary Medical Science. 70(10):1133-7.
· Salyers, A. A. and Whitt, D. D. (2005). Revenge of the Microbes: How bacterial resistance is undermining the antibiotic miracle. ASM Press, Washington, DC.
· Soltan-Dallal M. M., SharifiYazdi, M. K., Mirzaei, N.,Kalantar., E. (2014). Prevalence of Salmonella spp. in packed and unpacked red meat and chicken in South of Tehran. Jundishapur Journal Microbiology. 7(4): 9254-58.
· Tajbakhsh, M., YaghoobiAvini, M., Alikhajeh, J., Tajeddin, E., Rahbar, M., Eslami, P., et al., (2016). Emergence of blaCTX-M-15, blaTEM-169 and blaPER-1 extended-spectrum β-lactamase genes among different Salmonella entericaserovars fromhuman faecal samples. Infection Disease. 48(7):550-6.
· Tajbakhsh M, Yaghoobi Avini M, Ali Khajeh J, Alebouyeh M, Nazemalhosseini Mojarad E, Zali MR. (2012). Increased-Resistance Phenotype Resulted from Elevated β-Lactamase Enzyme Activity in Salmonella Clinical Isolates. Journal Isfahan Medical School. 30(178): 1-11. [In Persian]
· Wang, Y., Yang, B., Wu, Y., Zhang, Z., Meng, X., Xi, M., et al., (2015). Molecular characterization of Salmonella enterica serovar enteritidis on retail raw poultry in six provinces and two National cities in China. Food Microbiology. 46: 74-80.
· Wu, H., Wang, Y., Wu, Y., Qiao, J., Li, H., Zheng, S., et al., (2015). Emergence of β-Lactamases and extended-spectrum β-Lactamases (ESBLs) producing Salmonellain retail raw chicken in China. Foodborne Pathology Disease. 12(3): 228-235.
· Wu, H., Xia, X., Cui, Y, Hu, Y., Xi, M., Wang, X., et al., (2013). Prevalence of extended-spectrum β-Lactamase–producing Salmonella on retail chicken in six provinces and two national cities in the people’s republic of China. Journal Food Protection. 76(12): 2040–2044.
Yahyavi Firozabadi A, Sedighi khavidak S, Soltan Dallal M.M. (2016). The Molecular epidemiology evaluation and antibiotic resistance patterns of Salmonella isolated in food outbreaks in Yazd province by culture and PCR methods. Razi Journal Medical Science. 23(144): 1-8. [In Persian]
