تشخیص و تعیین فراوانی لیستریا مونوسیتوژنز در نمونه های مواد غذایی با بررسی مولکولی ژن hlyA
محورهای موضوعی :
میکروبیولوژی مواد غذایی
سوسن اشرفی
1
,
نسرین بهمنی
2
,
فاطمه کشاورزی
3
1 - کارشناس ارشد میکروبیولوژی، گروه زیست شناسی، واحد سنندج، دانشگاه آزاد اسلامی، سنندج، ایران
2 - استادیار مرکز تحقیقات زئونوز، پژوهشکده توسعه بهداشت، دانشگاه علوم پزشکی کردستان، سنندج، ایران
3 - دانشیار گروه زیست شناسی، واحد سنندج، دانشگاه آزاد اسلامی، سنندج، ایران
تاریخ دریافت : 1402/01/21
تاریخ پذیرش : 1402/04/17
تاریخ انتشار : 1402/07/01
کلید واژه:
مواد غذایی,
ژنA hly,
لیستریا مونوسیتوژنز,
چکیده مقاله :
مقدمه: لیستریولیزینo (LLO)) Listeriolysin O ( یک همولیزین فعال شده با سولفیدریل متصل به کلسترول است که توسط ژن hlyA لیستریا مونوسیتوژنز کدگذاری می شود. هدف از این مطالعه تشخیص و تعیین فراوانی لیستریا مونوسیتوژنز در نمونه های مواد غذایی با بررسی مولکولی ژن hlyA است. مواد و روش ها: مطالعه حاضر یک مطالعه مقطعی- توصیفی است. نمونه گیری از اواسط مهر ماه 1398تا پایان دی ماه همان سال به طول انجامید. در مجموع 165 نمونه انواع مختلف گوشت ، سبزیجات ، همبرگر و میگو مورد بررسی قرار گرفت. جمع آوری نمونه ها به صورت تصادفی از رستوران ها، اغذیه فروشی ها، مکان های تهیه غذای آماده و سوپر مارکت های مختلف انجام شد. کلیه نمونه های جمع آوری شده، با آزمایشات فنوتیپیکی و بیوشیمیایی مورد بررسی قرار گرفت و نمونه های مشکوک به لیستریا مونوسیتوژنز پس از استخراج DNA با روش جوشاندن با پرایمرهای احتصاصی برای ژن hlyA ارزیابی شد. یافته ها : بررسی های فنوتیپیک (کشت میکروبی و بررسی بیوشیمیایی) 18 نمونه آلوده به لیستریا مونوسیتوژنز را تایید کرد، از این تعداد بطریق PCR ژن hlyA 10(06/6% ) نمونه لیستریامونوسیتوز مثبت بود. شیوع آلودگی در کباب کوبیده 37/9 %، انواع گوشت مرغ 44/4 %، همبرگر محلی 66/16 % و کاهو 33/13 % بود. این یافته ها در مقایسه با دیگر مطالعات در برخی مواد غذایی(همبرگر محلی) شیوع بیشتری داشت و در بعضی مواد غذایی(گوشت مرغ) کمتر بود.نتیجه گیری: استفاده از روش بیوشیمیایی در تعیین فراوانی لیستریا مونوسیتوژنز در نمونه های مواد غذایی در شهرستان سنندج از درصد بالای مثبت کاذب برخوردار بود.
چکیده انگلیسی:
Introduction: Listeriolysin (LLO) is a sulfhydryl-activated hemolysin bound to cholesterol, that is encoded by the hlyA gene of Listeria monocytogenes. The aim of this study was to detect and determine the frequency of Listeria monocytogenes in food samples through molecular analysis of the hlyA gene.Materials and Methods: The present study was a descriptive cross-sectional assay conducted from the middle of October 2018 to the end of January of the same year. One hundred sixty-five samples of different types of meat, vegetables, hamburgers and shrimps were examined. Samples were randomly collected from restaurants, delicatessens, prepared food places, and different supermarkets. All samples were analyzed with phenotypic and biochemical tests. The samples suspected of Listeria monocytogenes, after DNA extraction, were evaluated by PCR with specific primers for the hlyA gene. Then the data and results were evaluated with using SPSS v.23 software and Excel software.Results: Phenotypic methods (microbial culture and biochemical investigation) confirmed 18 positive samples for Listeria monocytogenes, of which 10 (6.06%) samples were positive for Listeria monocytogenes by PCR of the hlyA gene. The rate of contamination in pounded kebab was 37.9%, chicken meat 4.4%, local hamburger 16.66% and lettuce 13.33%. Compared to other studies, these findings were more prevalent in some foods (e.g., local hamburger) and less in others (e.g., chicken meat).Conclusion: The use of biochemical methods for determining the frequency of Listeria monocytogenes in food samples in Sanandaj had a high percentage of false positives.
منابع و مأخذ:
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Chaitiemwong, N., Hazeleger, W.C., Beumer, R.R. & Zwietering, M.H. (2014). Quantification of transfer of Listeria monocytogenes between cooked ham and slicing machine surface. Food Control, 44, 177–184. https://doi.org/10.1016/j.foodcont.2014.03.056
Choi, W.S. & Hong, C. (2003). Rapid enumeration of Listeria monocytogenes in milk using competitive PCR. International Journal of Food Microbiology, 84, 79– 85. https://doi.org/10.1016/s0168-1605(02)00401-4
Di Pinto, A., Novello, L., Montemurro, F., Bonerba, E. & Tantillo G. (2010). Occurrence of Listeria monocytogenes in ready to eat foods from supermarkets in southern Italy. New Microbiology, 33, 249–252.
Esho, F. K., Enkhtuya, B., Kusumoto, A. & Kawamoto, K. (2013). Microbial Assessment and Prevalence of Foodborne Pathogens in Natural Cheeses in Japan. BioMed Research International, 20580, 1–6. https://doi.org/10.1155/2013/205801
Freitag, N.E., Rong, L. & Portnoy, D.A. (1993). Regulation of the prfA transcriptional activator of Listeria monocytogenes: multiple promoter elements contribute to intracellular growth and cell-to-cell spread. Infect Immune, 61 (6), 2537.
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Gamboa-Marín A., Buitrago S.M., Pérez-Pérez K., Mercado M.R., Poutou-Piñales R. & Carrascal-Camacho A. (2012). Prevalence of Listeria monocytogenes in pork-meat and other processed products from the Colombian swine industry. Rev MVZ Cordoba, 17, 2827–2833. https://doi.org/10.21897/rmvz.250
Greco, S., Tolli, R., Bossù, T., Flores Rodas, E. M., Di Giamberardino, F., Di Sirio, A., Vita, S., De 633 Angelis, V., Bilei, S., Sonnessa, M., Gattuso, A. & Lanni, L. (2014). Case of contamination 634 by Listeria monocytogenes in mozzarella cheese. Italian Journal of Food Safety, 3(1), 51-53.22. https://doi.org/10.4081/ijfs.2014.1708
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Hamidiyan, N., Salehi-Abargouei, A., Rezaei, Z., Dehghani-Tafti, R. & Akrami-Mohajeri, F. (2018). The prevalence of Listeria spp. Food contamination in Iran: A systematic review and meta-analysis. Food Research International, 107, 437-50. https://doi.org/10.1016/j.foodres.2018.02.038
Hellström, S., Laukkanen, R., Siekkinen, K.M., Jukka, R., Maijala, R. & Korkeala, H. (2010). Listeria monocytogenes contamination in pork can originate from farms. Journal of Food Protection, 73, 641–648. https://doi.org/10.4315/0362-028x-73.4.641
Hoelzer, K., Pouillot, R., Gallagher, D., Silverman, M.B., Kause, J. & Dennis, S. (2012). Estimation of Listeria monocytogenes transfer coefficients and efficacy of bacterial removal through cleaning and sanitation. International Journal of Food Microbiology, 157(2), 267-77. https://doi.org/10.1016/j.ijfoodmicro.2012.05.019
Henriques, A.R., Gama, L.T. & Fraqueza, M.J. (2017). Tracking Listeria monocytogenes contamination and virulence associated characteristic in the ready-to-eat meat-based food products industry according to hygiene level. International Journal of Food Microbiology, 242, 101–106. https://doi.org/10.1016/j.ijfoodmicro.2016.11.020
Jalali, M. & Abedi, D. (2008). Prevalence of Listeria speices in food products in Isfahan Iran. Int J Food Microbiol, 122(3), 336-40. https://doi.org/10.1016/j.ijfoodmicro.2007.11.082
Jami, S., Jamshidi, A. & Khanzadi, S. (2010). The presence of Listeria monocytogenes in raw milk samples in Mashhad, Iran. Iranian Journal of Veterinary Research, 11(4), 363-367. 22099/IJVR.2010.108
Kaur, S., Singh, R., Sran, M.K. & Gill, JP. (2018). Molecular characterization of Listeria monocytogenes in white meat samples from Punjab, India. Indian Journal of Animal Research, 52(11), 1635-41. https://doi.org/10.18805/ijar.b-3414
Kačániová, M., Kluz, M., Petrová, J., Mellen, M. & Kunová, S. (2015). Incidence of Listeria monocytogenes in meat product samples by real time PCR. Modern Chemistry & Applications, 3, 1–5. https://doi.org/10.4172/2329-6798.1000155
Kurpas, M., Wieczorek, K. & Osek, J. (2018). Ready-to-eat meat products as a source of Listeria monocytogenes. Journal of Veterinary Research, 62(1),49-55. https://doi.org/10.2478/jvetres-2018-0007
Kanuganti, S.R., Wesley, I.V., Reddy, P.G., McKean, J. & Hurd, H.S. (2002). Detection Listeria monocytogenes in pigs and pork. Journal of Food Protection, 65, 1470–1474. https://doi.org/10.4315/0362-028x-65.9.1470
Liu, D. (2006). Identification, subtyping and virulence determination of Listeria monocytogenes, an important foodborne pathogen. Journal of Medical Microbiology, 55(6), 645-59. https://doi.org/10.1099/jmm.0.46495-0
Montero, D., Bodero, M., Riveros, G., Lapierre, L., Gaggero, A., Vidal, R.M. &Vidal, M. (2015). Molecular epidemiology and genetic diversity of Listeria monocytogenes isolates from a wide variety of ready-to-eat foods and their relationship to clinical strains from listeriosis outbreaks in Chile. Front Microbiology, 6, 1–8. https://doi.org/10.3389/fmicb.2015.00384
Norowzi, J., Moradi, B. & Shafiee, M. (2013). Detection of actA gene in Listeria monocytogenes isolated from dairy products. Journal of Microbial World, 6(3), 246-252. [In Persian]
Osaili, T., Alaboudi, A. & Nesiar, E. (2011). Prevalence of Listeria spp. and antibiotic susceptibility of Listeria monocytogenes isolated from raw chicken and ready-to-eat chicken products in Jordan. Food Control, 22(3), 586-590. https://doi.org/10.1016/j.foodcont.2010.10.008
Orndorff, P.E., Hamrick, T.S., Smoak, I.W. & Havell, E.A. (2006). Host and bacterial factors in listeriosis pathogenesis. Veterinary Microbiology, 16, 114(1-2), 1-5. https://doi.org/10.1016/j.vetmic.2005.12.003
Pintado, C. M. B., Oliveira, A., Pampulha, M. E. & Ferreira, M. A. S. (2005). Prevalence and characterization of Listeria monocytogenes isolated from soft cheese. Food Microbiology, 22(1), 79–85. https://doi.org/10.1016/j.fm.2004.04.004
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Perrin, M., Bremer, M. & Delamare, C. (2007). Fatal microbiological methods, 71(2),13.
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Almeida, P.F. & Almeida, R.C.C. (2000). A PCR protocol using inl gene as a target for specific detection of Listeria monocytogenes. Food Control, 11(2), 97-10. https://doi.org/10.1016/s0956-7135(99)00067-5
(2016). European Centre for Disease Prevention and Control (ECDC): The European Union summary report on trends and sources of zoonoses, zoonotic agents, and food-borne outbreaks in 2015. EFSA Journal, 14. https://doi.org/10.2903/j.efsa.2016.4634
Akhondzadeh Basati, A., Misaghi, A., Salehi Zahraei, T. & Kamkar, A. (2006). Bacterial pathogens in fresh smoked and salted Irainian fish. Food Control, 17(3), 183-188. https://doi.org/10.1016/j.foodcont.2004.10.001
Baharvand, R. (2015). A gene in Listeria monocytogenes isolated from food. The Journal of Qazvin University of Medical Sciences, 19 (1), 24-31. [In Persian]
Bierne, H., Sabet, C., Personnic, N. & Cossart, P. (2007). Internalins: a complex family of leucine-richrepeat-containing proteins in Listeria monocytogenes. Microbes and Infection, 9, 1156-66. https://doi.org/10.1016/j.micinf.2007.05.003
Borović, B.B., Baltić, T., Lakićević, B., Janković, V., Mitrović, R. Jovanović, J. & Lilić S. (2014). Prevalence of Listeria monocytogenes in ready–to–eat food of animal origin. Meat Technology, 55, 117–122. https://doi.org/10.5937/tehmesa1402117b
Carpentier, B. & Cerf, O. (2011). Review — Persistence of Listeria monocytogenes in food industry equipment and premises. International Journal of Food Microbiology, 145, 1-8. https://doi.org/10.1016/j.ijfoodmicro.2011.01.005
Chaitiemwong, N., Hazeleger, W.C., Beumer, R.R. & Zwietering, M.H. (2014). Quantification of transfer of Listeria monocytogenes between cooked ham and slicing machine surface. Food Control, 44, 177–184. https://doi.org/10.1016/j.foodcont.2014.03.056
Choi, W.S. & Hong, C. (2003). Rapid enumeration of Listeria monocytogenes in milk using competitive PCR. International Journal of Food Microbiology, 84, 79– 85. https://doi.org/10.1016/s0168-1605(02)00401-4
Di Pinto, A., Novello, L., Montemurro, F., Bonerba, E. & Tantillo G. (2010). Occurrence of Listeria monocytogenes in ready to eat foods from supermarkets in southern Italy. New Microbiology, 33, 249–252.
Esho, F. K., Enkhtuya, B., Kusumoto, A. & Kawamoto, K. (2013). Microbial Assessment and Prevalence of Foodborne Pathogens in Natural Cheeses in Japan. BioMed Research International, 20580, 1–6. https://doi.org/10.1155/2013/205801
Freitag, N.E., Rong, L. & Portnoy, D.A. (1993). Regulation of the prfA transcriptional activator of Listeria monocytogenes: multiple promoter elements contribute to intracellular growth and cell-to-cell spread. Infect Immune, 61 (6), 2537.
https://doi.org/10.1128/iai.61.6.2537-2544.1993
Gamboa-Marín A., Buitrago S.M., Pérez-Pérez K., Mercado M.R., Poutou-Piñales R. & Carrascal-Camacho A. (2012). Prevalence of Listeria monocytogenes in pork-meat and other processed products from the Colombian swine industry. Rev MVZ Cordoba, 17, 2827–2833. https://doi.org/10.21897/rmvz.250
Greco, S., Tolli, R., Bossù, T., Flores Rodas, E. M., Di Giamberardino, F., Di Sirio, A., Vita, S., De 633 Angelis, V., Bilei, S., Sonnessa, M., Gattuso, A. & Lanni, L. (2014). Case of contamination 634 by Listeria monocytogenes in mozzarella cheese. Italian Journal of Food Safety, 3(1), 51-53.22. https://doi.org/10.4081/ijfs.2014.1708
Gómez, D., Pilar Iguácel, L., Rota, C., Carramiñana, J.J., Ariño, A. & Yangüela, J. (2015). Occurrence of Listeria monocytogenes in ready-to-eat meat products and meat processing plants in Spain. Foods, 4, 271–282. https://doi.org/10.3390/foods4030271
Gudbjörnsdóttir, B., Suihko, M. L., Gustavsson, P., Thorkelsson, G., Salo, S., Sjöberg, A. M., Niclasen, O. & Bredholt, S. (2004). The incidence of Listeria monocytogenes in meat, poultry and seafood plants in the Nordic countries. Food Microbial, 21, 217–222. https://doi.org/10.1016/s0740-0020(03)00012-1
Hamidiyan, N., Salehi-Abargouei, A., Rezaei, Z., Dehghani-Tafti, R. & Akrami-Mohajeri, F. (2018). The prevalence of Listeria spp. Food contamination in Iran: A systematic review and meta-analysis. Food Research International, 107, 437-50. https://doi.org/10.1016/j.foodres.2018.02.038
Hellström, S., Laukkanen, R., Siekkinen, K.M., Jukka, R., Maijala, R. & Korkeala, H. (2010). Listeria monocytogenes contamination in pork can originate from farms. Journal of Food Protection, 73, 641–648. https://doi.org/10.4315/0362-028x-73.4.641
Hoelzer, K., Pouillot, R., Gallagher, D., Silverman, M.B., Kause, J. & Dennis, S. (2012). Estimation of Listeria monocytogenes transfer coefficients and efficacy of bacterial removal through cleaning and sanitation. International Journal of Food Microbiology, 157(2), 267-77. https://doi.org/10.1016/j.ijfoodmicro.2012.05.019
Henriques, A.R., Gama, L.T. & Fraqueza, M.J. (2017). Tracking Listeria monocytogenes contamination and virulence associated characteristic in the ready-to-eat meat-based food products industry according to hygiene level. International Journal of Food Microbiology, 242, 101–106. https://doi.org/10.1016/j.ijfoodmicro.2016.11.020
Jalali, M. & Abedi, D. (2008). Prevalence of Listeria speices in food products in Isfahan Iran. Int J Food Microbiol, 122(3), 336-40. https://doi.org/10.1016/j.ijfoodmicro.2007.11.082
Jami, S., Jamshidi, A. & Khanzadi, S. (2010). The presence of Listeria monocytogenes in raw milk samples in Mashhad, Iran. Iranian Journal of Veterinary Research, 11(4), 363-367. 22099/IJVR.2010.108
Kaur, S., Singh, R., Sran, M.K. & Gill, JP. (2018). Molecular characterization of Listeria monocytogenes in white meat samples from Punjab, India. Indian Journal of Animal Research, 52(11), 1635-41. https://doi.org/10.18805/ijar.b-3414
Kačániová, M., Kluz, M., Petrová, J., Mellen, M. & Kunová, S. (2015). Incidence of Listeria monocytogenes in meat product samples by real time PCR. Modern Chemistry & Applications, 3, 1–5. https://doi.org/10.4172/2329-6798.1000155
Kurpas, M., Wieczorek, K. & Osek, J. (2018). Ready-to-eat meat products as a source of Listeria monocytogenes. Journal of Veterinary Research, 62(1),49-55. https://doi.org/10.2478/jvetres-2018-0007
Kanuganti, S.R., Wesley, I.V., Reddy, P.G., McKean, J. & Hurd, H.S. (2002). Detection Listeria monocytogenes in pigs and pork. Journal of Food Protection, 65, 1470–1474. https://doi.org/10.4315/0362-028x-65.9.1470
Liu, D. (2006). Identification, subtyping and virulence determination of Listeria monocytogenes, an important foodborne pathogen. Journal of Medical Microbiology, 55(6), 645-59. https://doi.org/10.1099/jmm.0.46495-0
Montero, D., Bodero, M., Riveros, G., Lapierre, L., Gaggero, A., Vidal, R.M. &Vidal, M. (2015). Molecular epidemiology and genetic diversity of Listeria monocytogenes isolates from a wide variety of ready-to-eat foods and their relationship to clinical strains from listeriosis outbreaks in Chile. Front Microbiology, 6, 1–8. https://doi.org/10.3389/fmicb.2015.00384
Norowzi, J., Moradi, B. & Shafiee, M. (2013). Detection of actA gene in Listeria monocytogenes isolated from dairy products. Journal of Microbial World, 6(3), 246-252. [In Persian]
Osaili, T., Alaboudi, A. & Nesiar, E. (2011). Prevalence of Listeria spp. and antibiotic susceptibility of Listeria monocytogenes isolated from raw chicken and ready-to-eat chicken products in Jordan. Food Control, 22(3), 586-590. https://doi.org/10.1016/j.foodcont.2010.10.008
Orndorff, P.E., Hamrick, T.S., Smoak, I.W. & Havell, E.A. (2006). Host and bacterial factors in listeriosis pathogenesis. Veterinary Microbiology, 16, 114(1-2), 1-5. https://doi.org/10.1016/j.vetmic.2005.12.003
Pintado, C. M. B., Oliveira, A., Pampulha, M. E. & Ferreira, M. A. S. (2005). Prevalence and characterization of Listeria monocytogenes isolated from soft cheese. Food Microbiology, 22(1), 79–85. https://doi.org/10.1016/j.fm.2004.04.004
Pan, Y., Breidt, F. Jr. & Kathariou S. (2006). Resistance of Listeria monocytogenes biofilms to sanitizing agents in simulated food processing environment. Applied and Environmental Microbiology, 72, 7711–7717. https://doi.org/10.1128/aem.01065-06
Perrin, M., Bremer, M. & Delamare, C. (2007). Fatal microbiological methods, 71(2),13.
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