بررسی میزان آلودگی بروسلا در شیرهای خام گوسفندی و اثر پالیده عاری از سلول (CFS) لاکتوباسیلوس اسیدوفیلوس روی تولید بیوفیلم آن
محورهای موضوعی :
بهداشت مواد غذایی
مسعود پرانداخته
1
(دانش‌آموخته دکتری عمومی دامپزشکی، واحد ارومیه، دانشگاه آزاد اسلامی، ارومیه، ایران)
مسلم نیریز نقدهی
2
(استادیار گروه بهداشت مواد غذایی، واحد ارومیه، دانشگاه آزاد اسلامی، ارومیه، ایران)
تاریخ دریافت : 1401/06/02
تاریخ پذیرش : 1401/07/25
تاریخ انتشار : 1401/06/01
کلید واژه:
بیوفیلم,
لاکتوباسیلوس اسیدوفیلوس,
بروسلا,
پالیده عاری از سلول (CFS),
شیر‌خام گوسفند,
مهاباد (ایران),
چکیده مقاله :
بروسلوز از بیماری‌های باکتریایی زئونوتیک مهم در انسان و حیوانات مهره‌دار است. باکتری‌های اسید‌لاکتیک و ترکیبات تولیدی آن‌ها به‌عنوان عوامل کنترل‌زیستی بالقوه در تشکیل بیوفیلم مطرح شده‌اند. تحقیق حاضر با هدف تعیین اثرات پالیده عاری از سلول (CFS) لاکتوباسیلوس اسیدوفیلوس روی تولید بیوفیلم جدایه‌های بروسلا از شیرهای خام گوسفندی شهرستان مهاباد انجام شد. در این تحقیق، 100 نمونه شیر گوسفندی بهصورت تصادفی از مناطق مختلف شهری و روستایی شهرستان مهاباد جمع‌آوری شدند. برای جداسازی بروسلا، ابتدا نمونه‌ها در آبگوشت بروسلا و سپس در آگار بروسلا با مکمل انتخابی کشت داده شدند؛ و برای شناسایی گونه‌های بروسلا، آزمایش‌های متابولیکی مرتبط انجام گردید. قابلیت تولید بیوفیلم جدایه‌های بروسلا و نیز فعالیت ضد‌بیوفیلم CFS لاکتوباسیلوس اسیدوفیلوس به روش سنجش میکروپلیت ارزیابی شد. یافته‌ها نشان داد که از مجموع 100 نمونه شیر، تعداد 3 نمونه (3 درصد) آلوده به بروسلا ملی‌تنسیس بودند. هم‌چنین هر سه جدایه دارای توانایی تشکیل بیوفیلم تشخیص داده شدند. دو جدایه، تولید کننده متوسط بیوفیلم و یک جدایه، مولد قوی بیوفیلم بودند. غلظت‌های مختلف CFS، بهصورت معنی‌داری (01/0˂p) تولید بیوفیلم جدایه‌ها را کاهش دادند. هم‌چنین غلظت 10 درصد CSF، بیشترین فعالیت ضد‌بیوفیلم را نشان داد. از یافته‌ها می‌توان استنباط نمود که آلودگی بروسلا در شیرهای خام گوسفندی منطقه مهاباد نسبتاً بالاست. اجرای کامل برنامه‌های غربالگری و مایه‌کوبی بروسلوز در گله‌های دامی منطقه ضروری است. هم‌چنین می‌توان نتیجه‌گیری نمود که CFS لاکتوباسیلوس اسیدوفیلوس دارای قابلیت حذف بیوفیلم بروسلا می‌باشد. بنابراین، CFS لاکتوباسیلوس اسیدوفیلوس به‌عنوان یک ترکیب ضد بیوفیلم می‌تواند در کنترل رشد بروسلا مطرح گردد.
چکیده انگلیسی:
Brucellosis is one of the most important zoonotic bacterial diseases in humans and vertebrate animals. Lactic acid bacteria (LAB) and their production compounds have been proposed as potential biocontrol agents in biofilm formation. The present research aimed to determine the effects of cell-free supernatant (CFS) from Lactobacillus acidophilus on the biofilm production of Brucella isolates from raw sheep milk of Mahabad city (Iran). In this study, 100 sheep milk samples were randomly collected from different urban and rural areas of Mahabad. To isolate Brucella, the samples were cultured in Brucella broth and then streaked on Brucella agar with the selective supplement, and to identify the species of Brucella, related metabolic tests were performed. The biofilm production capability of the isolates, as well as the antibiofilm activity of the CFS, were evaluated by the microplate assay method. The results showed that out of 100 milk samples, 3 samples (3%) are contaminated with Brucella melitensis. Also, all three isolates were biofilm-formers. Two isolates were recognized as moderate and one isolate as a strong biofilm-former. Different concentrations of CFS reduced the biofilm production of isolates significantly (P˂ 0.01). Also, the concentration of 10% of CSF showed the highest antibiofilm activity. From the findings, it can be concluded that Brucella contamination in raw sheep milk in the Mahabad region is relatively high. There is a need for a full performance of Brucellosis screening and vaccination programs in livestock herds of the region. It can also be concluded that the CFS of L. acidophilus can remove Brucella biofilm. Therefore, L. acidophilus CFS as an antibiofilm compound can be proposed to control the growth of Brucella.
منابع و مأخذ:
Abedi, A.S., Hashempour-Baltork, F., Alizadeh, A.M., Beikzadeh, S., Hosseini, H., Bashiry, M., et al. (2020). The prevalence of Brucella in dairy products in the Middle East region: a systematic review and meta-analysis. Acta Tropica, 202: 105241.
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Almirón, M.A., Roset, M.S. and Sanjuan, N. (2013). The aggregation of Brucella abortus occurs under microaerobic conditions and promotes desiccation tolerance and biofilm formation. Open Microbiology Journal, 7: 87–91.
Aminnezhad, S. and Kasra-Kermanshahi R. (2014). Antibiofilm activity of cell-free supernatant from Lactobacillus casei in Pseudomonas aeruginosa. Feyz (Journal of Kashan University of Medical Sciences), 18(1): 30-37. [In Persian]
Ashrafganjooyi, S.H., Saedadeli, N., Alamian, S., Khalili, M. and Shirazi, Z. (2017). Isolation and biotyping of Brucella from sheep and goats raw milk in southeastern Iran. Tropical Biomedicine, 34(3): 507–511.
Burmølle, M., Thomsen, T.R., Fazli, M., Dige, I., Christensen, L., Homøe, P., et al. (2010). Biofilms in chronic infections a matter of opportunity monospecies biofilms in multispecies infections. FEMS Immunology & Medical Microbiology, 59: 324-336.
Dadar, M., Alamian, S., Behrozikhah, A.M., Yazdani, F., Kalantari, A., Etemadi, A., et al. (2019). Molecular identification of Brucella species and biovars associated with animal and human infection in Iran. Veterinary Research Forum, 10 (4): 315-321.
Dadar, M., Shahalia, Y. and Whatmore, AM. (2019). Human brucellosis caused by raw dairy products: A review on the occurrence, major risk factors and prevention. International Journal of Food Microbiology, 292: 39-47.
Dang, H. and Lovell, C.R. (2015). Microbial surface colonization and biofilm development in marine environments. Microbiology and Molecular Biology Reviews, 80: 91-138.
Ebrahimi, M. and Zoghi, E., (2017). A review of the use of reduced-dose vaccine of melitensis Rev1 strain in adult sheep. Veterinary Journal, 115: 2-9. [In Persian]
Esmaeili, H., Ekhtiyarzadeh. H., Ebrahimzadeh, H., Partovi, R., Marhamati Khameneh, B., Hamedi, M., et al. (2012). Evaluation of the national sheep and goat brucellosis control program in Iran. Arak Medical University Journal, 14(6, Suppl 3): 9-20. [In Persian]
Esmaeili, H., Esmaeili, H. and Amiri, K., 2013. The effects of brucellosis vaccination in domestic animal on human brucellosis in Iran. Razi Journal of Medical Sciences, 20: 80-86. [In Persian]
Giordani, B., Parolin, C. and Vitali, B. (2021). Lactobacilli as anti-biofilm strategy in oral infectious diseases: a mini-review. Frontiers in Medical Technology, 3: 769172.
Godefroid, M., Svensson, M.V., Cambier, P., Uzureau1, S., Mirabella, A., De Bolle, X., et al. (2010). Brucella melitensis 16M produces a mannan and other extracellular matrix components typical of a biofilm. FEMS Immunology & Medical Microbiology, 59: 364–377.
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Jalilsood, T., Baradaran, A., Song, A.A.L., Ling Foo, H., Mustafa, S., Saad, W.Z., et al. (2015). Inhibition of pathogenic and spoilage bacteria by a novel biofilmforming Lactobacillus isolate: A potential host for the expression of heterologous proteins. Microbial Cell Factories, 14: 96.
Karim, G. and Farkhondeh, A. (1984). Milk and Public Health. 1st Edition, Markaz Nashr Daneshgahi, pp. 36-44. [In Persian].
Khelissa, S.O., Abdallah, M., Jama, C., Faille, C. and Chihib, N-E. (2017). Bacterial contamination and biofilm formation on abiotic surfaces and strategies to overcome their persistence. Journal of Materials and Environmental Sciences, 8(9): 3326-3346.
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Kumon, H., Ono, N., Iida, M. and Nickel, J.C. (1995). Combination effect of fosfomycin and ofloxacin against Pseudomonas aeruginosa growing in a biofilm. Antimicrobial Agents and Chemotherapy, 39 (5):1038-1044.
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Méndez-Vilas A. (2013). Microbial Pathogens and Strategies for Combating them: Science, Technology and Education. Formatex Research Center, Badajoz, pp. 42-51.
Mirnejad, R., Vahdati, A.R., Rashidiani, J., Erfani, M. and Piranfar, V. (2013). The antimicrobial effect of Lactobacillus casei culture supernatant against multiple drug resistant clinical isolates of Shigella Sonnei and Shigella Flexneri in vitro. Iranian Red Crescent Medical Journal, 5(2): 122-6.
Moslemi, E., Soltandalal, M.M., Beheshtizadeh, M.R., Taghavi, A., Kheiri Manjili., H.R, Mahmoudi Lamouki, R., et al. (2018). Detection of Brucella in dairy products by real-time PCR. Archives of Clinical Infectious Diseases, 13(1): e12673.
Nataraj, B.H., Ali, S.A., Behare, P. V. and Yadav, H. (2020). Postbiotics-parabiotics: the new horizons in microbial biotherapy and functional foods. Microbial Cell Factories, 19:168.
Naves, P., Del Prado, G., Huelves1, L., Gracia1, M., Ruiz1, V., Blanco, J., et al. (2008). Measurement of biofilm formation by clinical isolates of Escherichia coli is method-dependent. Journal of Applied Microbiology, 105:585-
Parsek, M.R. and Singh, P.K. (2003). Bacterial biofilms: An emerging link to disease pathogenesis. Annual review of Microbiology, 57: 677-701.
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Shafeie, B., Ahmadi, M. and Dastmalchi Saei, H. (2012). Diagnosis of Brucella abortus and Brucella melitensis in the milk of cattle and sheep in Kordestan province by polymerase chain reaction. Journal of Veterinary Microbiology. 8(2): 127-135. [In Persian]
Shakerian, A., Deo, P., Rahimi, E Shahjavan, A-R. and Khamesipour, F. (2015). Molecular detection of Brucella melitensis in sheep and goat milk in Iran. Tropical Journal of Pharmaceutical Research, 15 (5): 913-918.
Shokri, D., Khorasgani, M.R., Mohkam, M., Fatemi, S.M., Ghasemi, Y. and Taheri-Kafrani, A. (2018). The inhibition effect of lactobacilli against growth and biofilm formation of Pseudomonas aeruginosa. Probiotics and Antimicrobial Proteins, 10:34-
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Abedi, A.S., Hashempour-Baltork, F., Alizadeh, A.M., Beikzadeh, S., Hosseini, H., Bashiry, M., et al. (2020). The prevalence of Brucella in dairy products in the Middle East region: a systematic review and meta-analysis. Acta Tropica, 202: 105241.
Aguilar-Toalá, J.E, Garcia-Varela, R., Garcia, H.S., Mata-Haro, V., GonzálezCórdova, A.F., Vallejo-Cordoba, B., et al. (2018). Postbiotics: An evolving term within the functional foods field. Trends in Food Science and Technology, 75: 105–14.
Ait Ouali, F., Al Kassaa, I., Cudennec, B., Abdallah, M., Bendali, F., Sadoun, D., et al. (2014). Identification of lactobacilli with inhibitory effect on biofilm formation by pathogenic bacteria on stainless steel surfaces. International Journal of Food Microbiology, 191: 116-124.
Almirón, M.A., Roset, M.S. and Sanjuan, N. (2013). The aggregation of Brucella abortus occurs under microaerobic conditions and promotes desiccation tolerance and biofilm formation. Open Microbiology Journal, 7: 87–91.
Aminnezhad, S. and Kasra-Kermanshahi R. (2014). Antibiofilm activity of cell-free supernatant from Lactobacillus casei in Pseudomonas aeruginosa. Feyz (Journal of Kashan University of Medical Sciences), 18(1): 30-37. [In Persian]
Ashrafganjooyi, S.H., Saedadeli, N., Alamian, S., Khalili, M. and Shirazi, Z. (2017). Isolation and biotyping of Brucella from sheep and goats raw milk in southeastern Iran. Tropical Biomedicine, 34(3): 507–511.
Burmølle, M., Thomsen, T.R., Fazli, M., Dige, I., Christensen, L., Homøe, P., et al. (2010). Biofilms in chronic infections a matter of opportunity monospecies biofilms in multispecies infections. FEMS Immunology & Medical Microbiology, 59: 324-336.
Dadar, M., Alamian, S., Behrozikhah, A.M., Yazdani, F., Kalantari, A., Etemadi, A., et al. (2019). Molecular identification of Brucella species and biovars associated with animal and human infection in Iran. Veterinary Research Forum, 10 (4): 315-321.
Dadar, M., Shahalia, Y. and Whatmore, AM. (2019). Human brucellosis caused by raw dairy products: A review on the occurrence, major risk factors and prevention. International Journal of Food Microbiology, 292: 39-47.
Dang, H. and Lovell, C.R. (2015). Microbial surface colonization and biofilm development in marine environments. Microbiology and Molecular Biology Reviews, 80: 91-138.
Ebrahimi, M. and Zoghi, E., (2017). A review of the use of reduced-dose vaccine of melitensis Rev1 strain in adult sheep. Veterinary Journal, 115: 2-9. [In Persian]
Esmaeili, H., Ekhtiyarzadeh. H., Ebrahimzadeh, H., Partovi, R., Marhamati Khameneh, B., Hamedi, M., et al. (2012). Evaluation of the national sheep and goat brucellosis control program in Iran. Arak Medical University Journal, 14(6, Suppl 3): 9-20. [In Persian]
Esmaeili, H., Esmaeili, H. and Amiri, K., 2013. The effects of brucellosis vaccination in domestic animal on human brucellosis in Iran. Razi Journal of Medical Sciences, 20: 80-86. [In Persian]
Giordani, B., Parolin, C. and Vitali, B. (2021). Lactobacilli as anti-biofilm strategy in oral infectious diseases: a mini-review. Frontiers in Medical Technology, 3: 769172.
Godefroid, M., Svensson, M.V., Cambier, P., Uzureau1, S., Mirabella, A., De Bolle, X., et al. (2010). Brucella melitensis 16M produces a mannan and other extracellular matrix components typical of a biofilm. FEMS Immunology & Medical Microbiology, 59: 364–377.
Gomez, N.C., Ramiro, J.M., Quecan, B.X.V. and De Melo Franco, B.D.G. (2016). Use of potential probiotic lactic acid bacteria (lab) biofilms for the control of Listeria monocytogenes, Salmonella typhimurium, and Escherichia coli O157:H7 biofilms formation. Frontiers in Microbiology, 7: 863.
Halling, S.M., and Young, E.J. (2001). Brucella, In: Hu, Y. H., Pierson, M.D., and Gorham, J.R. (Editors), Foodborne Disease Handbook Volume 1: Bacterial Pathogens. Marcel Dekker, Inc. pp. 77-82.
Institute of Standards and Industrial Research of Iran. (ISIRI), (2016). Microbiology of food and animal feeding stuffs –General requirements and guidance for microbiological examinations. 1st edition, ISIRI No. 9899. [In Persian]
Institute of Standards and Industrial Research of Iran. (ISIRI), (2020). Microbiology of Food Chain- Complete Method for Isolation and Identification of Brucella 1st edition, ISIRI No. 19153. [In Persian].
Jalilsood, T., Baradaran, A., Song, A.A.L., Ling Foo, H., Mustafa, S., Saad, W.Z., et al. (2015). Inhibition of pathogenic and spoilage bacteria by a novel biofilmforming Lactobacillus isolate: A potential host for the expression of heterologous proteins. Microbial Cell Factories, 14: 96.
Karim, G. and Farkhondeh, A. (1984). Milk and Public Health. 1st Edition, Markaz Nashr Daneshgahi, pp. 36-44. [In Persian].
Khelissa, S.O., Abdallah, M., Jama, C., Faille, C. and Chihib, N-E. (2017). Bacterial contamination and biofilm formation on abiotic surfaces and strategies to overcome their persistence. Journal of Materials and Environmental Sciences, 8(9): 3326-3346.
Khiralla, G.M., Mohamad, E.A.H., Farag, A.G. and Elhariry, H. (2015). Antibiofilm effect of Lactobacillus pentosus and Lactobacillus plantarum cell-free supernatants against some bacterial pathogens. Journal of Biotechnology Research, 6: 86-95.
Koohestani, M., Moradi, M., Tajik, H. and Badali, A. (2018). Effects of cell-free supernatant of Lactobacillus acidophilus LA5 and Lactobacillus casei 431 against planktonic form and biofilm of Staphylococcus aureus. Veterinary Research Forum, 9(1): 301-306.
Kumon, H., Ono, N., Iida, M. and Nickel, J.C. (1995). Combination effect of fosfomycin and ofloxacin against Pseudomonas aeruginosa growing in a biofilm. Antimicrobial Agents and Chemotherapy, 39 (5):1038-1044.
Lopes, L.B., Nicolino, R. and Haddad, J.P.A. (2017). Brucellosis - Risk Factors and Prevalence: A Review. The Open Veterinary Science Journal, 4: 72-84.
Marouf, A.S Hanifian, S. and Shayegh, J. (2021). Prevalence of Brucella in raw milk and artisanal cheese tested via real-time qPCR and culture assay. International Journal of Food Microbiology, 347: 109192.
Méndez-Vilas A. (2013). Microbial Pathogens and Strategies for Combating them: Science, Technology and Education. Formatex Research Center, Badajoz, pp. 42-51.
Mirnejad, R., Vahdati, A.R., Rashidiani, J., Erfani, M. and Piranfar, V. (2013). The antimicrobial effect of Lactobacillus casei culture supernatant against multiple drug resistant clinical isolates of Shigella Sonnei and Shigella Flexneri in vitro. Iranian Red Crescent Medical Journal, 5(2): 122-6.
Moslemi, E., Soltandalal, M.M., Beheshtizadeh, M.R., Taghavi, A., Kheiri Manjili., H.R, Mahmoudi Lamouki, R., et al. (2018). Detection of Brucella in dairy products by real-time PCR. Archives of Clinical Infectious Diseases, 13(1): e12673.
Nataraj, B.H., Ali, S.A., Behare, P. V. and Yadav, H. (2020). Postbiotics-parabiotics: the new horizons in microbial biotherapy and functional foods. Microbial Cell Factories, 19:168.
Naves, P., Del Prado, G., Huelves1, L., Gracia1, M., Ruiz1, V., Blanco, J., et al. (2008). Measurement of biofilm formation by clinical isolates of Escherichia coli is method-dependent. Journal of Applied Microbiology, 105:585-
Parsek, M.R. and Singh, P.K. (2003). Bacterial biofilms: An emerging link to disease pathogenesis. Annual review of Microbiology, 57: 677-701.
Quinn, P.J., Carter, M.E., Markey, B. and Carter, G.R. (1999). Clinical Veterinary Microbiology. Elsevier Limited, pp. 261-268.
Shafeie, B., Ahmadi, M. and Dastmalchi Saei, H. (2012). Diagnosis of Brucella abortus and Brucella melitensis in the milk of cattle and sheep in Kordestan province by polymerase chain reaction. Journal of Veterinary Microbiology. 8(2): 127-135. [In Persian]
Shakerian, A., Deo, P., Rahimi, E Shahjavan, A-R. and Khamesipour, F. (2015). Molecular detection of Brucella melitensis in sheep and goat milk in Iran. Tropical Journal of Pharmaceutical Research, 15 (5): 913-918.
Shokri, D., Khorasgani, M.R., Mohkam, M., Fatemi, S.M., Ghasemi, Y. and Taheri-Kafrani, A. (2018). The inhibition effect of lactobacilli against growth and biofilm formation of Pseudomonas aeruginosa. Probiotics and Antimicrobial Proteins, 10:34-
Shunmugaperumal, T. (2010). Biofilm Eradication and Prevention: A Pharmaceutical Approach to Medical Device Infections. Hoboken, USA: John Wiley & Sons, pp. 3-36.
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