اثربخشی نایسین ریزپوشانی شده با سدیم آلژینات و کیتوزان بر تشکیل بیوفیلم در لیستریا مونوسایتوجنز
محورهای موضوعی :
بهداشت مواد غذایی
سجاد باباخانی
1
,
فرزانه حسینی
2
,
پرویز پاکزاد
3
,
مریم بی خوف تربتی
4
1 - دانشجوی دکتری گروه میکروبیولوژی، دانشکده علوم زیستی، واحد تهران شمال، دانشگاه آزاد اسلامی، تهران، ایران
2 - گروه میکروبیولوژی، دانشکده علوم زیستی، واحد تهران شمال، دانشگاه آزاد اسلامی، تهران، ایران
3 - استاد گروه میکروبیولوژی، دانشکده علوم زیستی، واحد تهران شمال، دانشگاه آزاد اسلامی، تهران، ایران
4 - استادیارگروه زیست‌شناسی، دانشکده علوم، واحد شهر ری، دانشگاه آزاد اسلامی، تهران، ایران
تاریخ دریافت : 1401/04/14
تاریخ پذیرش : 1401/05/29
تاریخ انتشار : 1401/03/01
کلید واژه:
پروبیوتیک,
مواد غذایی,
لیستریوزیس,
تهاجم سلولی,
میکروکپسوله‌سازی,
ویرولانس,
چکیده مقاله :
لیستریا مونوسایتوجنز عامل لیستریوزیس منتقله از طریق غذا است و بقای طولانی از طریق تشکیل بیوفیلم دارد. هدف این مطالعه بررسی اثر رقت IU/ml 102 نایسین و نایسین میکروکپسوله شده با کیتوزان و سدیم آلژینات بر تشکیل بیوفیلم و تغییر سطح بیان ژن‌های مرتبط با بیوفیلم در لیستریا مونوسایتوجنز بود. لیستریا مونوسایتوجنز ATCC 19115، سروتایپ b4 به‌وسیله نایسین IU/ml 102 و نایسین میکروکپسوله با کیتوزان و سدیم آلژینات تیمار شد. اثربخشی نایسین بر بقای سلولی از طریق محاسبه جذب نوری برآورد شد. پس از اثبات وجود ژن‌های prfA، sigB و agrA، اثر نایسین IU/ml 102 بر روی بیوفیلم لیستریا مونوسایتوجنز به روش میکروتیتر پلیت بررسی شد. تغییر بیان ژن‌های prfA، sigB و agrA با استفاده از روش ریل تایم (qRT-PCR) واکاوی شد. میزان مهار بیوفیلم لیستریا مونوسایتوجنز در دمای 37 درجه سلسیوس و 5/5 pHتوسط نایسین 57 درصد بود. بیشترین مهار بیوفیلم مربوط به استفاده هم‌زمان نایسین IU/ml 102 با کیتوزان و سدیم آلژینات در دمای 37 درجه سلسیوس و 5/5 pHبود. ریزپوشانی با کیتوزان و سدیم آلژینات توانست اثرگذاری نایسین در مهار بیوفیلم را به 76 درصد برساند (0001/0p<). نایسین IU/ml 102 موجب کاهش بیان ژن‌های prfA، sigB و agrA به‌ترتیب به میزان 313/2-، 808/2- و 453/1- برابر نسبت به کنترل شد (p<0.0001). نایسین فعالیت مهاری قابل‌توجهی در برابر بیوفیلم لیستریا مونوسایتوجنز داشت. نایسین میکروکپسوله شده با کیتوزان و سدیم آلژینات دارای یک اثربخشی فوق‌العاده در مهار بیوفیلم لیستریا مونوسایتوجنز بود. نایسین IU/ml 102 می‌تواند موجب کاهش بیان برخی ژن‌های درگیر در تشکیل بیوفیلم لیستریا مونوسایتوجنز شود.
چکیده انگلیسی:
Listeria monocytogenes is a foodborne pathogen the cause of listeriosis that has long-term survival due to biofilm formation. This study aimed to investigate the efficacy of nisin 102 IU/ml, encapsulated nisin with chitosan and sodium alginate on biofilm formation and changing the expression level of genes related to biofilm formation L. monocytogenes ATCC 19115 (serotype 4b) was treated by nisin 102 IU/ml and microencapsulated nisin with chitosan and sodium alginate solutions. The effectiveness of nisin on cell survival was estimated by calculating the optical absorbance. After proving the presence of prfA, sigB, and agrA genes, the effect of 102 IU/ml nisin dilution on L. monocytogenes biofilm was investigated by the microtiter plate method. The expression level of prfA, sigB, and agrA genes was analyzed using real-time PCR methods (qRT-PCR). The inhibition rate of biofilm formation in L. monocytogenes by nisin 102 IU/ml was 57% at 37°C and pH 5.5. The maximum inhibition of biofilm formation was related to the simultaneous use of nisin 102 IU/ml with chitosan and sodium alginate at 37°C and pH 5.5. Microencapsulated nisin with chitosan and sodium alginate can increase its effectiveness in preventing the biofilm formation of L. monocytogenes to 76% (p<0.0001). Nisin 102 IU/ml decreased the expression level of prfA, sigB, and agrA by -2.313, -2.808, and -1.453-fold compared to the control (p<0.0001). Nisin only had significant inhibitory activity against L. monocytogenes biofilm. Microencapsulated nisin with chitosan and sodium alginate had high efficacy in preventing the biofilm formation of L. monocytogenes.Listeria monocytogenes is a foodborne pathogen the cause of listeriosis that has long-term survival due to biofilm formation. This study aimed to investigate the efficacy of nisin 102 IU/ml, encapsulated nisin with chitosan and sodium alginate on biofilm formation and changing the expression level of genes related to biofilm formation L. monocytogenes ATCC 19115 (serotype 4b) was treated by nisin 102 IU/ml and microencapsulated nisin with chitosan and sodium alginate solutions. The effectiveness of nisin on cell survival was estimated by calculating the optical absorbance. After proving the presence of prfA, sigB, and agrA genes, the effect of 102 IU/ml nisin dilution on L. monocytogenes biofilm was investigated by the microtiter plate method. The expression level of prfA, sigB, and agrA genes was analyzed using real-time PCR methods (qRT-PCR). The inhibition rate of biofilm formation in L. monocytogenes by nisin 102 IU/ml was 57% at 37°C and pH 5.5. The maximum inhibition of biofilm formation was related to the simultaneous use of nisin 102 IU/ml with chitosan and sodium alginate at 37°C and pH 5.5. Microencapsulated nisin with chitosan and sodium alginate can increase its effectiveness in preventing the biofilm formation of L. monocytogenes to 76% (p<0.0001). Nisin 102 IU/ml decreased the expression level of prfA, sigB, and agrA by -2.313, -2.808, and -1.453-fold compared to the control (p<0.0001). Nisin only had significant inhibitory activity against L. monocytogenes biofilm. Microencapsulated nisin with chitosan and sodium alginate had high efficacy in preventing the biofilm formation of L. monocytogenes.
منابع و مأخذ:
Abebe, E., Gugsa, G. and Ahmed, M. (2020). Review on Major Food-Borne Zoonotic Bacterial Pathogens. Journal of Tropical Medicine, 2020: 4674235.
Balciunas, E.M., Martinez, F.A.C., Todorov, S.D., de Melo Franco, B.D.G., Converti, A. and de Souza Oliveira, R.P. (2013). Novel biotechnological applications of bacteriocins: a review. Food Control, 32(1): 134-142.
Brandelli, A. (2012). Nanostructures as promising tools for delivery of antimicrobial peptides. Mini reviews in medicinal chemistry, 12(8): 731-741.
Cabeça, T.K., Pizzolitto, A.C. and Pizzolitto, E.L. (2012). Activity of disinfectants against foodborne pathogens in suspension and adhered to stainless steel surfaces. Brazilian Journal of Microbiology, 43(3): 1112-1119.
Chaturongakul, S., Raengpradub, S., Wiedmann, M. and Boor, K.J. (2008). Modulation of stress and virulence in Listeria monocytogenes. Trends in Microbiology, 16(8): 388-396.
Fahim, H.A., Khairalla, A.S. and El-Gendy, A.O. (2016). Nanotechnology: a valuable strategy to improve bacteriocin formulations. Frontiers in Microbiology, 7: 1385.
Feng, L. and Mumper, R.J. (2013). A critical review of lipid-based nanoparticles for taxane delivery. Cancer letters, 334(2): 157-175.
Galie, S., Garcia-Gutierrez, C., Miguelez, E.M., Villar, C.J. and Lombo, F. (2018). Biofilms in the food industry: health aspects and control methods. Frontiers in Microbiology, 9: 898.
Good, J.A., Andersson, C., Hansen, S., Wall, J., Krishnan, K.S., Begum, A. et al., (2016). Attenuating Listeria monocytogenes virulence by targeting the regulatory protein PrfA. Cell Chemical Biology, 23(3): 404-414.
Gray, J., Chandry, P.S., Kaur, M., Kocharunchitt, C., Fanning, S., Bowman, J.P. et al., (2021). Colonisation dynamics of Listeria monocytogenes strains isolated from food production environments. Scientific Reports, 11(1): 12195.
Hernandez-Milian, A. and Payeras-Cifre, A. (2014). What is new in listeriosis? BioMed Research International, 358051.
Kiral, E., kacmaz, E., Bozan, G., Arslanoglu, O., Kilic, O. and Dinleyici, E.C. (2021). 261. A rare case of meningitis and symptomatic hydrocephalus by Listeria monocytogenes in dermatomyositis: A Case Report. Open Forum Infectious Diseases, 8(Supplement_1): S237-S238.
Li, M. (2020), Exploring the connection between acid exposure and virulence in Listeria monocytogenes, Utah State University.
Liu, Y., Wu, L., Han, J., Dong, P., Luo, X., Zhang, Y. et al., (2020). Inhibition of biofilm formation and related gene expression of Listeria monocytogenes in response to four natural antimicrobial compounds and sodium hypochlorite. Frontiers in Microbiology, 11: 617473.
Livak, K.J. and Schmittgen, T.D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods, 25(4): 402-408.
Lopes, N.A., Pinilla, C.M.B. and Brandelli, A. (2019). Antimicrobial activity of lysozyme-nisin co-encapsulated in liposomes coated with polysaccharides. Food Hydrocolloids, 93: 1-9.
Maresca, D., De Prisco, A., La Storia, A., Cirillo, T., Esposito, F. and Mauriello, G. (2016). Microencapsulation of nisin in alginate beads by vibrating technology: Preliminary investigation. LWT-Food Science and Technology, 66: 436-443.
Martinez, R.C.R., Alvarenga, V.O., Thomazini, M., Fávaro-Trindade, C.S. and de Souza Sant'Ana, A. (2016). Assessment of the inhibitory effect of free and encapsulated commercial nisin (Nisaplin®), tested alone and in combination, on Listeria monocytogenes and Bacillus cereus in refrigerated milk. LWT-Food Science and Technology, 68: 67-75.
Mauriello, G., De Luca, E., La Storia, A., Villani, F. and Ercolini, D. (2005). Antimicrobial activity of a nisin‐activated plastic film for food packaging. Letters in Applied Microbiology, 41(6): 464-469.
Pinheiro, J., Lisboa, J., Pombinho, R., Carvalho, F., Carreaux, A., Brito, C. et al., (2018). MouR controls the expression of the Listeria monocytogenes Agr system and mediates virulence. Nucleic Acids Research, 46(18): 9338-9352.
Pinilla, C.M.B., Stincone, P. and Brandelli, A. (2021). Proteomic analysis reveals differential responses of Listeria monocytogenes to free and nanoencapsulated nisin. International Journal of Food Microbiology, 346: 109170.
Pisano, M.B., Fadda, M.E., Melis, R., Ciusa, M.L., Viale, S., Deplano, M. et al., (2015). Molecular identification of bacteriocins produced by Lactococcus lactis dairy strains and their technological and genotypic characterization. Food Control, 51: 1-8.
Prombutara, P., Kulwatthanasal, Y., Supaka, N., Sramala, I. and Chareonpornwattana, S. (2012). Production of nisin-loaded solid lipid nanoparticles for sustained antimicrobial activity. Food Control, 24(1-2): 184-190.
Rahmati, F. (2017). Characterization of Lactobacillus, Bacillus and Saccharomyces isolated from Iranian traditional dairy products for potential sources of starter cultures. AIMS Microbiology, 3(4): 815.
Rahmati, F. (2020). Microencapsulation of Lactobacillus acidophilus and Lactobacillus plantarum in Eudragit S100 and alginate chitosan under gastrointestinal and normal conditions. Applied Nanoscience, 10(2): 391-399.
Ramaswamy, V., Cresence, V.M., Rejitha, J.S., Lekshmi, M.U., Dharsana, K., Prasad, S.P. et al., (2007). Listeria-review of epidemiology and pathogenesis. Journal of Microbiology Immunology and Infection, 40(1): 4.
Rossi, M.L., Paiva, A., Tornese, M., Chianelli, S. and Troncoso, A. (2008). Listeria monocytogenes outbreaks: a review of the routes that favor bacterial presence. Revista Chilena de Infectologia: organo oficial de la Sociedad Chilena de Infectologia, 25(5): 328-335.
Salama, Y., Chennaoui, M., Sylla, A., Mountadar, M., Rihani, M. and Assobhei, O. (2016). Characterization, structure, and function of extracellular polymeric substances (EPS) of microbial biofilm in biological wastewater treatment systems: a review. Desalination and Water Treatment, 57(35): 16220-16237.
Sen, C. and Ray, P.R. (2019). Biopreservation of dairy products using bacteriocins. Indian Food Industry, 1: 51-60.
Stincone, P., Miyamoto, K.N., Timbe, P.P.R., Lieske, I. and Brandelli, A. (2020). Nisin influence on the expression of Listeria monocytogenes surface proteins. Journal of Proteomics, 226: 103906.
Urban, P., Jose Valle-Delgado, J., Moles, E., Marques, J., Diez, C. and Fernandez-Busquets, X. (2012). Nanotools for the delivery of antimicrobial peptides. Current Drug Targets, 13(9): 1158-1172.
van der Veen, S. and Abee, T. (2010). Importance of SigB for Listeria monocytogenes static and continuous-flow biofilm formation and disinfectant resistance. Applied and Environmental Microbiology, 76(23): 7854-7860.
Were, L.M., Bruce, B., Davidson, P.M. and Weiss, J. (2004). Encapsulation of nisin and lysozyme in liposomes enhances efficacy against Listeria monocytogenes. Journal of Food Protection, 67(5): 922-927.
Zhao, X., and Kuipers, O.P. (2021). Synthesis of silver-nisin nanoparticles with low cytotoxicity as antimicrobials against biofilm-forming pathogens. Colloids and Surfaces B: Biointerfaces, 206: 111965.
Zhao, X., Zhao, F., Wang, J. and Zhong, N. (2017). Biofilm formation and control strategies of foodborne pathogens: food safety perspectives. RSC Advances, 7(58): 36670-36683.
Zohri, M., Alavidjeh, M.S., Haririan, I., Ardestani, M.S., Ebrahimi, S.E.S., Sani, H.T. et al., (2010). A comparative study between the antibacterial effect of nisin and nisin-loaded chitosan/alginate nanoparticles on the growth of Staphylococcus aureus in raw and pasteurized milk samples. Probiotics and Antimicrobial Proteins, 2(4): 258-266.
Zohri, M., Shafiee Alavidjeh, M., Mirdamadi, S.S., Behmadi, H., Hossaini Nasr, S.M., Eshghi Gonbaki, S. et al., (2013). Nisin‐loaded chitosan/alginate nanoparticles: A hopeful hybrid biopreservative. Journal of Food Safety, 33(1): 40-49.
_||_
Abebe, E., Gugsa, G. and Ahmed, M. (2020). Review on Major Food-Borne Zoonotic Bacterial Pathogens. Journal of Tropical Medicine, 2020: 4674235.
Balciunas, E.M., Martinez, F.A.C., Todorov, S.D., de Melo Franco, B.D.G., Converti, A. and de Souza Oliveira, R.P. (2013). Novel biotechnological applications of bacteriocins: a review. Food Control, 32(1): 134-142.
Brandelli, A. (2012). Nanostructures as promising tools for delivery of antimicrobial peptides. Mini reviews in medicinal chemistry, 12(8): 731-741.
Cabeça, T.K., Pizzolitto, A.C. and Pizzolitto, E.L. (2012). Activity of disinfectants against foodborne pathogens in suspension and adhered to stainless steel surfaces. Brazilian Journal of Microbiology, 43(3): 1112-1119.
Chaturongakul, S., Raengpradub, S., Wiedmann, M. and Boor, K.J. (2008). Modulation of stress and virulence in Listeria monocytogenes. Trends in Microbiology, 16(8): 388-396.
Fahim, H.A., Khairalla, A.S. and El-Gendy, A.O. (2016). Nanotechnology: a valuable strategy to improve bacteriocin formulations. Frontiers in Microbiology, 7: 1385.
Feng, L. and Mumper, R.J. (2013). A critical review of lipid-based nanoparticles for taxane delivery. Cancer letters, 334(2): 157-175.
Galie, S., Garcia-Gutierrez, C., Miguelez, E.M., Villar, C.J. and Lombo, F. (2018). Biofilms in the food industry: health aspects and control methods. Frontiers in Microbiology, 9: 898.
Good, J.A., Andersson, C., Hansen, S., Wall, J., Krishnan, K.S., Begum, A. et al., (2016). Attenuating Listeria monocytogenes virulence by targeting the regulatory protein PrfA. Cell Chemical Biology, 23(3): 404-414.
Gray, J., Chandry, P.S., Kaur, M., Kocharunchitt, C., Fanning, S., Bowman, J.P. et al., (2021). Colonisation dynamics of Listeria monocytogenes strains isolated from food production environments. Scientific Reports, 11(1): 12195.
Hernandez-Milian, A. and Payeras-Cifre, A. (2014). What is new in listeriosis? BioMed Research International, 358051.
Kiral, E., kacmaz, E., Bozan, G., Arslanoglu, O., Kilic, O. and Dinleyici, E.C. (2021). 261. A rare case of meningitis and symptomatic hydrocephalus by Listeria monocytogenes in dermatomyositis: A Case Report. Open Forum Infectious Diseases, 8(Supplement_1): S237-S238.
Li, M. (2020), Exploring the connection between acid exposure and virulence in Listeria monocytogenes, Utah State University.
Liu, Y., Wu, L., Han, J., Dong, P., Luo, X., Zhang, Y. et al., (2020). Inhibition of biofilm formation and related gene expression of Listeria monocytogenes in response to four natural antimicrobial compounds and sodium hypochlorite. Frontiers in Microbiology, 11: 617473.
Livak, K.J. and Schmittgen, T.D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods, 25(4): 402-408.
Lopes, N.A., Pinilla, C.M.B. and Brandelli, A. (2019). Antimicrobial activity of lysozyme-nisin co-encapsulated in liposomes coated with polysaccharides. Food Hydrocolloids, 93: 1-9.
Maresca, D., De Prisco, A., La Storia, A., Cirillo, T., Esposito, F. and Mauriello, G. (2016). Microencapsulation of nisin in alginate beads by vibrating technology: Preliminary investigation. LWT-Food Science and Technology, 66: 436-443.
Martinez, R.C.R., Alvarenga, V.O., Thomazini, M., Fávaro-Trindade, C.S. and de Souza Sant'Ana, A. (2016). Assessment of the inhibitory effect of free and encapsulated commercial nisin (Nisaplin®), tested alone and in combination, on Listeria monocytogenes and Bacillus cereus in refrigerated milk. LWT-Food Science and Technology, 68: 67-75.
Mauriello, G., De Luca, E., La Storia, A., Villani, F. and Ercolini, D. (2005). Antimicrobial activity of a nisin‐activated plastic film for food packaging. Letters in Applied Microbiology, 41(6): 464-469.
Pinheiro, J., Lisboa, J., Pombinho, R., Carvalho, F., Carreaux, A., Brito, C. et al., (2018). MouR controls the expression of the Listeria monocytogenes Agr system and mediates virulence. Nucleic Acids Research, 46(18): 9338-9352.
Pinilla, C.M.B., Stincone, P. and Brandelli, A. (2021). Proteomic analysis reveals differential responses of Listeria monocytogenes to free and nanoencapsulated nisin. International Journal of Food Microbiology, 346: 109170.
Pisano, M.B., Fadda, M.E., Melis, R., Ciusa, M.L., Viale, S., Deplano, M. et al., (2015). Molecular identification of bacteriocins produced by Lactococcus lactis dairy strains and their technological and genotypic characterization. Food Control, 51: 1-8.
Prombutara, P., Kulwatthanasal, Y., Supaka, N., Sramala, I. and Chareonpornwattana, S. (2012). Production of nisin-loaded solid lipid nanoparticles for sustained antimicrobial activity. Food Control, 24(1-2): 184-190.
Rahmati, F. (2017). Characterization of Lactobacillus, Bacillus and Saccharomyces isolated from Iranian traditional dairy products for potential sources of starter cultures. AIMS Microbiology, 3(4): 815.
Rahmati, F. (2020). Microencapsulation of Lactobacillus acidophilus and Lactobacillus plantarum in Eudragit S100 and alginate chitosan under gastrointestinal and normal conditions. Applied Nanoscience, 10(2): 391-399.
Ramaswamy, V., Cresence, V.M., Rejitha, J.S., Lekshmi, M.U., Dharsana, K., Prasad, S.P. et al., (2007). Listeria-review of epidemiology and pathogenesis. Journal of Microbiology Immunology and Infection, 40(1): 4.
Rossi, M.L., Paiva, A., Tornese, M., Chianelli, S. and Troncoso, A. (2008). Listeria monocytogenes outbreaks: a review of the routes that favor bacterial presence. Revista Chilena de Infectologia: organo oficial de la Sociedad Chilena de Infectologia, 25(5): 328-335.
Salama, Y., Chennaoui, M., Sylla, A., Mountadar, M., Rihani, M. and Assobhei, O. (2016). Characterization, structure, and function of extracellular polymeric substances (EPS) of microbial biofilm in biological wastewater treatment systems: a review. Desalination and Water Treatment, 57(35): 16220-16237.
Sen, C. and Ray, P.R. (2019). Biopreservation of dairy products using bacteriocins. Indian Food Industry, 1: 51-60.
Stincone, P., Miyamoto, K.N., Timbe, P.P.R., Lieske, I. and Brandelli, A. (2020). Nisin influence on the expression of Listeria monocytogenes surface proteins. Journal of Proteomics, 226: 103906.
Urban, P., Jose Valle-Delgado, J., Moles, E., Marques, J., Diez, C. and Fernandez-Busquets, X. (2012). Nanotools for the delivery of antimicrobial peptides. Current Drug Targets, 13(9): 1158-1172.
van der Veen, S. and Abee, T. (2010). Importance of SigB for Listeria monocytogenes static and continuous-flow biofilm formation and disinfectant resistance. Applied and Environmental Microbiology, 76(23): 7854-7860.
Were, L.M., Bruce, B., Davidson, P.M. and Weiss, J. (2004). Encapsulation of nisin and lysozyme in liposomes enhances efficacy against Listeria monocytogenes. Journal of Food Protection, 67(5): 922-927.
Zhao, X., and Kuipers, O.P. (2021). Synthesis of silver-nisin nanoparticles with low cytotoxicity as antimicrobials against biofilm-forming pathogens. Colloids and Surfaces B: Biointerfaces, 206: 111965.
Zhao, X., Zhao, F., Wang, J. and Zhong, N. (2017). Biofilm formation and control strategies of foodborne pathogens: food safety perspectives. RSC Advances, 7(58): 36670-36683.
Zohri, M., Alavidjeh, M.S., Haririan, I., Ardestani, M.S., Ebrahimi, S.E.S., Sani, H.T. et al., (2010). A comparative study between the antibacterial effect of nisin and nisin-loaded chitosan/alginate nanoparticles on the growth of Staphylococcus aureus in raw and pasteurized milk samples. Probiotics and Antimicrobial Proteins, 2(4): 258-266.
Zohri, M., Shafiee Alavidjeh, M., Mirdamadi, S.S., Behmadi, H., Hossaini Nasr, S.M., Eshghi Gonbaki, S. et al., (2013). Nisin‐loaded chitosan/alginate nanoparticles: A hopeful hybrid biopreservative. Journal of Food Safety, 33(1): 40-49.
