Relationship between biofilm production and virulence factors in Yersinia enterocolitica strains isolated from red meat in Shahrekord city
Subject Areas : Food microbiology
najmeh molavi
1
*
,
Manouchehr Momeni-Shahraki
2
,
Hossein Khodabandeh shahraki
3
1 - azad shahrekord
2 - گروه بهداشت مواد غذایی، دانشکده دامپزشکی، دانشگاه آزاد شهرکرد، ایران.
3 -
Keywords: Meat, Virulence genes, Yersinia enterocolitica.,
Abstract :
Yersinia enterocolitica is a gram-negative bacterium that belongs to the Enterobacteriaceae family. This bacterium is a food pathogenic bacterium that is mainly transmitted by contaminated meat, milk and water. Unsanitary meat is one of the main sources of Yersinia enterocolitica infection to humans. The present study was conducted with the aim of investigating the frequency of virulence genes and the relationship between biofilm production and virulence factors in Yersinia enterocolitica isolates isolated from meat supplied to Shahrekord market. 384 samples of red meat were randomly collected from meat supply stores in Shahrekord city and analyzed by biochemical and molecular methods in order to identify Yersinia enterocolitica. The isolated microbial strains were investigated in order to investigate the biofilm production ability by microtiter plate method and the virulence genes were investigated by PCR test. From a total of 384 red meat samples, Yersinia enterocolitica was isolated from 65 samples (16.92%). Contamination was reported in beef (27.69%), mutton (33.84%) and veal (38.64%). 35 isolates (53.85%) showed strong biofilm reaction, 20 isolates (30.77%) showed moderate biofilm reaction and 10 isolates (15.38%) showed weak biofilm reaction. The frequencies of inv, ail, yadA, ystA and virF genes were reported as: 83%, 41.53%, 43% 33.84% and 29.23%, respectively. In the statistical analysis with Chi-square test, a significant relationship was observed between virulence genes and strong biofilm reaction.
Considering the importance of red meat in the transmission of Yersinia enterocolitica to humans, it is recommended to follow the hygiene principles and not to consume contaminated meat, to use the PCR test in order to accurately identify the contamination in food.
1- Alenizi, D., Ringwood, T., Redhwan, A., Bouraha, B., Wren, B. W., Prentice, M. (2016). All Yersinia enterocolitica are pathogenic: Virulence of phylogroup 1 Y. enterocolitica in a Galleria mellonella infection model. Microbiology (Reading(: 162 (8): 1379-1387.
2- Altrock, A., Seinige, D., Kehrenberg, C. (2015). Yersinia enterocolitica isolates from wild boars hunted in lower saxony, Germany. Applied and Environmental Microbiology, 81(14), 4835–4840.
3- Bancerz-Kisiel, A., and Lipczynska-Ilczuk, K. (2021). Evaluation of the correlation between the mRNA expression Levels of ystA and ymoA genes in Y. enterocolitica strains with different enterotoxic properties. Pathogens. 10 (9): 1136. 1150.
4- Bancerz-Kisiel, A., Szczerba-Turek, A., Platt-Samoraj, A., Michalczyk, M., Szweda, W. (2018). Characterisation of ail-positive Yersinia enterocolitica of different biotypes using HRMA. International Journal of Food Microbiology. 269: 46–51.
5- Bari, M. L., Hossain, M. A., Isshiki, K. Ukuku, D. (2011). Behavior of Yersinia enterocolitica in foods. Journal of Pathogens. 54 (2): 1-12.
6- Bernardino-Varo, L., Quinones-Ramírez, E. I., Fernandez, F.J.V azquez-Salinas, C. (2013). Prevalence of Yersinia enterocolitica in raw cow’s milk collected from stables of Mexico City. Journal of Food Protection. 76(4): 694–698.
7- Bhagat, N. and Virdi, J.S. (2011). The enigma of Yersinia enterocolitica biovar 1A. Critical Reviews in Microbiology. 37(1): 25–39.
8- Bonardi, S., Bruini, I., D’Incau, M., Van Damme, I., Carniel, E., Br´emont, S. (2016). Detection, seroprevalence and antimicrobial resistance of Yersinia enterocolitica and Yersinia pseudotuberculosis in pig tonsils in Northern Italy. International Journal of Food Microbiology. 235 (5): 125–132.
9- Bonardi, S., Paris, A., Bassi, L., Salmi, F., Bacci, C., Riboldi, E., et al. (2010). Detection, semiquantitative enumeration, and antimicrobial susceptibility of Yersinia enterocolitica in pork and chicken meats in Italy. Journal of Food Protection. 73(10): 1785–1792.
10- Bottone, E. J. (2015). Yersinia enterocolitica: Revisitation of an enduring human pathogen. Clinical Microbiology Newsletter. 37(1): 387-405.
11- رحیم زاده، زراسوندی، ع.، 1382، جستجوی یرسینیا انتروکولیتیکا در شیرخام و پاستوریزه در استان چهارمحال و بختیاری ،دانشکده دامپزشکی، پایان نامه برای دریافت درجه دکترای حرفه‌ای دامپزشکی از دانشگاه آزاد اسلامی، واحد شهرکرد، ص. 48-8.
12- شریف زاده، ع و اخوان، م.، 1383، جداسازی یرسینیا انتروکولیتیکا و لیستریا مونوسایتوژنز از شیرهای خام پاستوریزه عرضه شده در سطح فروشگاه‌های استان چهارمحال و بختیاری، فصلنامه علوم صنایع غذایی ایران، دوره 1، شماره 1.
13- سالک مقدم، ع.، فروهش، هـ.، انصاری، ج.، 1380، بررسی آلودگی میکروبی در نمونه‌های پنیر غیر پاستوریزه در مقایسه با پنیرهای پاستوریزه و تأثیر مقادیر مختلف نمک اضافه شده به پنیر بر روی باکتری‌های بیماری‌زای آلوده کننده، مجله دانشگاه علوم پزشکی ایران، سال ششم، شماره 25.
14- سلطان دلان، م.م.، ایزدپور، ف.، خلیفه قلی، م.، خاتمی مقدم، م. و حجازی، س.ح.، 1382، بررسی یرسینیا در گوشت‌های ماهی قرمز و مرغ عرضه شده در قصابی‌ها و مرغ فروشی‌های تحت نظارت دانشگاه علوم پزشکی تهران، ص18-10.
15- سلطان دلال، م.، خرمی زاده، م.، متین، ف. 1385، بررسی روش‌های تشخیصی یرسینیا انتروکولیتیکا، مجله دانشگاه علوم پزشکی تهران،ص. 10-5.
16- سلطان دلال، م.، اخی، م.ت.، پرتوآذر، ع. 1387، مطالعه یرسینیا انتروکولیتیکا در اسهال‌های حاد کودکان زیر چهارده سال در مرکز طبی اطفال بیمارستان امام خمینی تهران، مجله پزشکی، دانشگاه علوم پزشکی تبریز، دوره 30، شماره 4، صفحه 52.-49.
17- فرشچیان، م.، مهدوی، س.، سروش، م.، 1388، بررسی میزان آلودگی گوشت‌های قرمز توزیع شده در شهر تبریز به یرسینیا انتروکولیتیکا، مجله دامپزشکی دانشگاه آزاد اسلامی، شماره 11، سال چهارم.
18- قیاسیان، م، قندهاری ،ف.، 1388، باکتری شناسی تشخیصی، انتشارات کنکاش، چاپ اول، ص.50-4.
19- کرمانشاهی، ک.، 1381، مکانیسم مولکولی بیماری‌زایی باکتری‌ها، انتشارات جهاد دانشگاهی، واحد اصفهان، چاپ اول. ص. 50-30.
20- مکفرسون، ج.، گیرمولر، س.، 1390، PCR (واکنش زنجیره‌ای پلی‌مراز)، ترجمه زینی، غ.، میرزایی،ع.، الهیان، ف.، انتشارات آییژ، ص.50-5.
21- ملک زاده، ف.، 1381، میکروب‌شناسی، انتشارات دانشگاه تهران، ص 217 و 325-322.
22- ملنیک و آدلیرک، 2007، میکروبیولوژی پزشکی، ترجمه خزعلی، م. و ضیایی، م.، ویراست بیست و چهارم، انتشارات حیان، جلد اول، ص.361-303.
23- یادمان، ر.، 1375، بررسی میزان آلودگی شیرخام و پاستوریزه به یرسینیا انتروکولیتیکا در منطقه اهواز و حومه، دانشکده دامپزشکی، پایان نامه برای دریافت درجه‌ی دکترای حرفه‌ای دامپزشکی از دانشگاه شهید چمران، پایان نامه دکترای حرفه‌ای، صفحه 90تا50.
24- هاریسون، 1375، اصول طب داخلی هاریسون، ترجمه ابراهیمی، ر. و داداش زاده، چاپ دوم، انتشارات حیان، صفحه 528 تا 534
25-Aulisiso, C.C, Hi11, W.E., Stanfield, J.T., 1983, Evaluation of virulence factor testing and Characteristics of pathogenicity in Yersinia enterocolitica infect, Immun, 40:300-335
.26-Bancerz, A., Szczerba, A., platt – samoraj, A., 2010, Application of multiplex PCR for the evaluation of the occurrence of yestA, yestB, ystC and ymoA genes in Yersinia enterocolitica,1,55:33-37.
27-Bancerz, A., Szczreba, A., platt, A., 2011, Isolation, Biotyping and serotping of Yersinia entercoliica, Bull Vet Inst pulawy.55: 39-43.
28-Bhagat, N., vird:, J.S, 2007, Distribution of virulrnce. Associated in Yersinia enterocolitica biovar 1A correlates with clonal groups and not the source of isolation, FEMS Microbiology letters, 226: 83-183.
29-Biase, A.M., Petrone, G., Conte, M.P., 2000, Infection of human entrocyte-like cells with rotaviruse enhances in vasiveness of Yersinia enterocolitia and Y.pseudotuber culosic, J. Med. Microbial, P. 49: 897-904.
30- Blais, B. W. and Phillippe, L., N., 1995, Compavative analysis of yadA and ail polymerase chain reaction- methods for virulent Yersinia enterocolitica, Food control, 6: 211-214.
31-Bottone, E.J., 1999, Yersina enterocolitica overview and epidemiologic correlates, Microbes infect, 1(4): 323-30.
32-Bottone, E.J., 1997, Yersinia enterocolitica the charismacotinuses clinical microbiolog Reviews, 10: 257-276.
33-Brham, E.P., 1981, The betalactam a antibiotics, Jour. Scientific A merican,.37, (4):76-86
34-Bronfine, D.R, Kroli, L.R., 2003, Yersinia entercolitica fection, 18(1):1-10
35-Capita, R., 2002, Incidence and pathogencity of Yersina Spp. From poultry in spain, food Microbiol, 19: 295-301.
36-Carter, G.R., 1994. Clinical Veterinary Microbiology, Wolfe pub: 234-5.
37-Cheyne, B., Van Dayke, M., An evaluation of methods for the isolation of Yersinia enterocolitica from surface waters in the Grand River watershed, to the university of waterloo in fulfillment of the thesis requirement.
38-Collee, J.G., Duguw, J.P., Fraser, A.G., 1998, Paractical Medical Microbiology, 3th edition, Longman Singapore publisher (LTD), pp. 226-6, 536-7.
39-Cumpansion. C.E,. 2009., Characterization of some Yersina Spp using the AP1 20E system, Medicin Veterinara, Vol 12 (1):275-282
40-Dwight, C., Hirch, M., 1999, Veterinary Microbiology, Black Cell Pup, pp. 28-3, 102-103.
41-Fredksson-Ahoma, M., Korkeala, H., 2003, Low occurrence of pathogenic Yersinia enterocolitica clinical food and environmental samples, climicro Rev : 220-229.
42- Fukushima, H., Shimizu, S. and Inatsu, Y. 2011, Yersinia enterocolitica and Yersina pseudotuberculosis detection in food, Journal of pathogens, Article 10735308, doi: 10. 4061-2011/735308.
43-Havens, J., Monthgomery, M., greecnon, E., 2003, Pathogenic Yersinia.entercolitca DNA is detected in gastrointestional malakplakia, Modern pathology, 14:1200a.
44- Ibraham, A., Rea, M., 1991, isolation of Yersinia enterocolitica and related species from red meat and milk, J. Food.Sci, 56:1524-1526.
.45- Johnson, J., 1998, Isolation and Identification of pathogene Yersinia enterocolitica from meat and pultry products, microbiology laboratory Guid book, 3 Edition.
46-Juliana P. Falcao, Deise P. Falcao, Andre Pitondo-Silva, Ana Carolina Malaspina and Marcelo Brocchi,2006,Molecular typing and virulence markers of Yersinia enterocolitica strains from human, animal and food origins isolated between 1968 and 2000 in Brazil ,Journal of Medical Microbiology, 55, 1539–1548.
47- Kapperud, G., Vardund, T., skyerve, E. Hornes, E., 1993, Detection of pathogenic Yersinia enterocolitica in foods and water by immunogenettic sepayation, nested polymerase chain reaction, and colorimetric detection of amplified DNA, Appi Environ Microbiol, 59 (9): 2938-2977.
48-Khorramizadeh, solta – Dallal, M., Savifor, F.,2007, Assessment of ail gene marker amplicon for molecular characteri2ation of pathogenic Yersinai enterocolitica in food samples collected in Iran, Iranian. J. publ. Health.:8-15.
49-LambertZ, S.T., Nilsson, C., Hallanoun, S., Lindblad,M.,2008, Real – time PCR method for detection of pathogenic Yersinia enterocolitica in food, Applied and Environmental Microbiology, 74(19):6060-6067
50-Lamps, L.W., Madhusudhan, K.T., Greenson, J.K.,. pierce, R.H., Massol, N.A., 2001, The Role of Yersinia enterocolitica and Yersinia pseudotuberculosis in granulomatousappendicitis, American. Journal .Surgical .Pathology, 25: 508-515.
51-Lewin. A., Strauch, E., Hertwig., S., Hoffmann, B., Nattrmann, H., Apple, B., 1996, Comparison of plasmid of strains of Yersinia enterocolitica biovar 1A with the virulence plasmid of a pathogenic strain, 285: 52-630.
52-Macdonald, E., Heier, B.T., 2011, Yersinia enterocolitica O:9 infections associated with bagged salad mix in Norway, Ferbrury to April 2011, Euro sarve: 11, 16(19): 19866.
53-Mauro, A., Lagana, P., Brouno, G., Micali, M., 2008, Isolation of Yersinai enterocolitica biotype 1A from raw meat products, J.PREV. VED .HYG, 49: 75-18.
54-Murry, p., 1998, Medical Microbiology. 3 th wishington DC Asempres pub, pp: 240-315.
55-Najdanski, H.,Iteman, I., Carnile, E, 1994, Efficienet subyping of pathogenic Yersinia enterocolitica strains by pulshed – fleid gel electero phoresis, Journal .Clinical. Micro :2913-20.
56-Nesbakken, T., Iversen, T., Lium, ., 2007, Pig herds free from human pathogenic Yersinai enterocolitica, Emery.Infect. Dis, 13 (12): 186-40
57-Niskanen, T., waldnstrom, J., Fredriksson – ahomaa, M., 2003, Vir F positive Yersinia pseudotuberculosis and Yersinia enterocolitica found in migratory birds in Sweden, Applied and environmental Microbiology, 69: 4670-4675.
58- Okwori, A.E., Agina, S.E., Olabode, A.O., 2005, Feacal carriage of Yesrina species in pigs sheep and poultry on display for sale in Vom and Bukuru aresas of jos south local Government Area (LGA). Plateau stat, J.Microbiol:387-230.
59- Ostroff, S.,1995,Yersiniaas an emerging infection Epidemiologic aspects of yersiniosis,Microbiol.Immunol,13:5-10.
60-Pham, J.N. Bell, S.M. Martin, L., Camiel, E., 2000, The beta – lactamases and beta-lactam antibiotic susceptibility of Yesinia enterocolitica, Antimicrob Chemother, 46(6): 951-1.
61-Quinn, P., Carter, M., Markey, B., Catrert, S., 1994, Clinical veterinary Microbiology wolf, pp. 234-236.
62-Robert, F., Hore, G., 1991, Basic medical microbiology 4th edition, Robert F. Body and Brayan G. Hoert pub, pp. 493-7.
63-Sabina, Y., Rahman, A., Chandra, R., Montet, D., 2011, Yersinia enterocolitica: mode of Transmission, Molecular insights of virulence, and pathogensis of Infection, Journal of pathogens, ID 429069, dio: 10. 4061/2011/4290069.
64-Salmah, A.A. and Makki, S., 1991, Insidence of Yersinai enterocolitica in some food environmental samples in Saudi Arabic, J .king. Saud. Univ, 3(2): 91-100
65-Sedgwick, A., Tilt, N.C., 1970, biochemical and serological charactenstics of a Yersinia enterocolitica isolate, Appled Microbiology: 383-384.
66-Sharifi, M., Soltan-Dellal, 2011, incidence and antibiotins suseptibilites of Yersinia enterocolitica and other Yersinia Species recovers from meat and chiken in Tehran, Iran, African Journal of Microbiology Research 5(18): 2649-2653.
67-Simonova., J. Valzerova, M., Stenhauserova, I., 2007, Detection of pathogenic Yersinai enterocolitica serotype O:3 by biochemical, serological, and PCR methods, Czech J food SCi, 25: 214-220.
68-Sirkem, B., 2002, The presence of Yersinia enterocolitica and other Yersina species in ground beef in Aydil Turkey.Turk.J vet.Anim sci.28:489-495
69-Soltan-Dallal, M., Chita2, M., 1996., Prevalence of Yersinai enterocolitica in peariatric dysentery, Facalty .Mesi. J, 1996:1: 4246.
70-Stock, I., Wiedemann, B., 1999, An invitrostudy of the antimicrobial susceptibilities of Yersinai enterocolitica and definition of a database, The journal of antimicrobial Chemotherapy, 43: 37-45.
71-Thisted lambertz, S., Danielsson – Them, N., 2005, Idenrification and characterization of pathogenic Yersinia enterocolitica isolates by PCR , Applies and Environmental Microbiology, 71: 3674-3681.
72-Thoemer, P., Kingombe, C., Eissig-Choisa, B., 2003, PCR Detection of virulence genes in Yersinia enterocolitica and Yersinia pseudotuberclosis and investingation of virulence gene distribution, applied and Enviromental Microbiology, 69: 1810-1816.
73-Velin, D., 1984, Enterotoxin production by Yersinia enterocolitica in food samples, Acta Microbiol Hung, 31(1): 43-8.
74-Vidon, D., Delmas. , C., 1981. Incidence of Yersinia enterocolitica in raw milk in eastern france, Appl. Environ. Microbial, 1981: 41(2): 355-9.
75-Warren, S.M., Young, G.X., 2005,An amino ytermila secretion signal is requires for YPLA export by the Yas, YSC, and flagellar type III secrection systems of Yersinai enterocolitica biovar 1B, J.Bacteriol, 187 (17): 6075-83.
76-Wannet, W.J., Ressinj, M., 2001, Detection of pathogenic Yersinia enterocolitica by a rapid and sensitive duplex PCR assay, Journal of Clinical Microbiology, 39(2): 4483-7786.
77-Wauters, G., kansol, K., and Janssens, M., 1987. Revised biogroply scheme of Yersinia enterocolitica , contrib. Microbiol. Immunol, 9: 14-21.
78-Weunats, V., Jadot. V., Denoel, P.A, Tibor., A., Letesson, J.J., 1996, Detection of Yersinia enterocolitica sergoup O: 3 by a PCR methos, J. Clin. Microbial, 1996 34(5): 1224-7.
79-Yerma, N.k. and Misra, 1984, Characterization of enterotoxin produced by four Yersinia enterocolitica strains of pig origin, Mesival Microbiology, 50(4), Dol: 10. 1007/BF00399650.
80-Zheng, H., Sun, Y., 2008, Investigation of virulence gence in slinical isolates of Yersinia enterocolitica, Medical Microbiology 53: 368-374.
81- رحیم زاده، زراسوندی، ع.، 1382، جستجوی یرسینیا انتروکولیتیکا در شیرخام و پاستوریزه در استان چهارمحال و بختیاری ،دانشکده دامپزشکی، پایان نامه برای دریافت درجه دکترای حرفه‌ای دامپزشکی از دانشگاه آزاد اسلامی، واحد شهرکرد، ص. 48-8.
82- Sharifi Yazdi, M. and Soltan-Dellal. (2011). Incidence and antibiotins suseptibilites of Yersinia enterocolitica and other Yersinia Species recovers from meat and chiken in Tehran, Iran, African Journal of Microbiology Research. 5(18): 2649-2653.
83- Thoemer, P., Kingombe, C., Eissig-Choisa, B. (2003). PCR Detection of virulence genes in Yersinia enterocolitica and Yersinia pseudotuberclosis and investingation of virulence gene distribution, applied and Enviromental Microbiology. 69: 1810-1816.
84- Thisted lambertz, S., Danielsson – Them, N. (2005). Idenrification and characterization of pathogenic Yersinia enterocolitica isolates by PCR. Applies and Environmental Microbiology. 71: 3674-3681.
85- Zexun L, Xiumin S, Jin Ch, Mingqian Qa, Huanjing Sh, Qian Zh, Jinlei Zh, Jun Y, Shenghui C, Fengqin L, Chengqian F, Zixin P, Baowei Y. (2022). Prevalence, bio-serotype, antibiotic susceptibility and genotype of Yersinia enterocolitica and other Yersinia species isolated from retail and processed meats in Shaanxi Province, China. Food Science and Technology. 168 ;113962
85- Piotr Ł., Klaudia K.K., Agata BK. 2023. Isolation, characterization and antimicrobial resistance of Yersinia enterocolitica from Polish cattle and their carcasses. BMC Veterinary Research. 19:143-152.
86- Thisted Lambert S, Danielsson-Tham ML. Identification and characterization of pathogenic Yersinia enterocolitica isolates by PCR and pulsed-field gel electrophoresis.
Applied and Environmental Microbiology. 2005: 71(7): 3674-3681
87- Sirkem, B., 2002, The presence of Yersinia enterocolitica and other Yersina species in ground beef in Aydil Turkey.Turk.J vet.Anim sci.28:489-495.
88- Naves P., Del Prado G., Huelves L., Gracia M., Ruiz V., Blanco J. Soriano F. (2008). Measurement of biofilm formation by clinical isolates of Escherichia coli is method‐dependent. Journal of Applied Microbiology. 105 (2); 585-590.