hfbHealth, Food & BiotechnologyHealth, Food & Biotechnology2712-7648РОСБИОТЕХ10.36107/hfb.2019.i4.s281hfb-49Research ArticleПИТАНИЕFOODДетекция жизнеспособных некультивируемыхклеток микроорганизмов в курином фаршеDetection of Viable but Nоnculturable Microbial Cells in Chicken MinceАбдуллаеваА. М.AbdullaevaA. M.

Абдуллаева Асият Мухтаровна

125080, Москва, Волоколамское шоссе, дом 11

Asiyat M. Abdullaeva

11 Volokolamskoe highway, Moscow, 125080, Russian Federation

abdullaevaam@mgupp.ruБлинковаЛ. П.BlinkovaL. P.

Блинкова Лариса Петровна

105064, город Москва, Малый Казенный переулок, дом 5а

Larisa P. Blinkova

5а, Maliy Kazenniy pereulok, Moscow, 105064, Russan Federation

b.larus@mail.ruУшаБ. В.UshaB. V.

Уша Борис Вениаминович

125080, Москва, Волоколамское шоссе, дом 11

Boris V. Usha

11 Volokolamskoe highway, Moscow, 125080, Russian Federation

vet-san-dekanat@yandex.ruВалитоваР. К.ValitovaR. K.

Валитова Румия Камилевна

125080, Москва, Волоколамское шоссе, дом 11

105064, город Москва, Малый Казенный переулок, дом 5а

Rumiya K. Valitova

11 Volokolamskoe highway, Moscow, 125080, Russian Federation

5а, Maliy Kazenniy pereulok, Moscow, 105064, Russan Federation

rumiya.valitova@gmail.comПахомовЮ. Д.PakhomovYu. D.

Пахомов Юрий Дмитриевич

105064, город Москва, Малый Казенный переулок, дом 5а

Yuriy D. Pakhomov

5а, Maliy Kazenniy pereulok, Moscow, 105064, Russan Federation

yury-pakhomov@yandex.ruМитрофановаД. Б.MitrofanovaD. B.

Митрофанова Дарья Борисовна

125080, Москва, Волоколамское шоссе, дом 11

105064, город Москва, Малый Казенный переулок, дом 5а

Daria B. Mitrofanova

11 Volokolamskoe highway, Moscow, 125080, Russian Federation

5а, Maliy Kazenniy pereulok, Moscow, 105064, Russan Federation

dar.gushina@yandex.ruФГБОУ ВО «Московский государственный университет пищевых производств»РоссияMoscow State University of Food ProductionRussian FederationФГБНУ НИИ вакцин и сывороток им. И.И. МечниковаРоссияI.I. Mechnikov Research Institute of Vaccines and SeraRussian FederationФГБОУ ВО «Московский государственный университет пищевых производств»; ФГБНУ НИИ вакцин и сывороток им. И.И. МечниковаРоссияMoscow State University of Food Production; I.I. Mechnikov Research Institute of Vaccines and SeraRussian Federation201905052021142638Copyright © Абдуллаева А.М., Блинкова Л.П., Уша Б.В., Валитова Р.К., Пахомов Ю.Д., Митрофанова Д.Б., 20202020Абдуллаева А.М., Блинкова Л.П., Уша Б.В., Валитова Р.К., Пахомов Ю.Д., Митрофанова Д.Б.Abdullaeva A.M., Blinkova L.P., Usha B.V., Valitova R.K., Pakhomov Y.D., Mitrofanova D.B.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.hfb-journal.ru/jour/article/view/49

В связи с потенциальной возможностью существования в пищевых продуктах биоопасных жизнеспособных некультивируемых клеток (ЖНК) патогенных микроорганизмов, которые могут формироваться под действием различных факторов, актуальными для предотвращения заражения мясных продуктов являются детекция и определение условий образования в них ЖНК разных бактерий-контаминантов. В работе проведен поиск ЖНК в курином фарше в условиях реального времени и при экспериментальном заражении продукции Staphylococcus aureus. Для выявления ЖНК в курином фарше (производство региональной птицефабрики) в 1 г продукта определяли совокупное количество микробов, численность выросших бактериальных колоний (КОЕ), содержание живых (мертвых) клеток с помощью коммерческого набора флуоресцентных красителей. Повторное изучение проводили через 5 ч инкубации тестируемых образцов фарша при комнатной температуре. В пробах куриного фарша на 4-й день после его изготовления обнаружены ЖНК в количестве более 99 % от всех выявленных живых клеток. Через 5 часов инкубации фарша при комнатной температуре бактерии-контаминанты увеличили численность КОЕ/г в 22,5 раза, но количество ЖНК сохранялось на уровне более 99 % от числа жизнеспособных бактерий. При искусственном заражении той же серии фарша бульонной культурой S. aureus 209 P в стадии логарифмического роста количество ЖНК в образце 0 час составило 97,33 %, а через 5 ч их численность повысилась до 99,99 %. Вероятно, во внесенной культуре стафилококка в стадии активного размножения не происходило образования ЖНК, что снизило вначале их количество в пробе. После 5 ч инкубации переход бактерий в состояние ЖНК ускорился, возможно, из-за неблагоприятных для популяции клеток условий (изменение трофического субстрата, температуры, pH и т.д.). Полученные данные являются экспериментальным подтверждением наличия в продукции из куриного мяса малоизученных ЖНК бактерий, не вырастающих на традиционных питательных средах, и показывающих ложноотрицательный результат при традиционной микробиологической экспертизе. Ввиду биоопасности таких скрытых дормантных клеток целесообразно предусмотреть регламентированное тестирование пищевой продукции на наличие ЖНК.

There is a potential for existence of hazardous viable but nonculturable (VBNC) cells of pathogenic microorganisms in foodstuffs that can be formed under the influence of various factors. Their detection and determination of conditions for formation of VBNC cells of various bacteria are relevant for preventing contamination of meats. This was the aim of the present study. The search was conducted for VBNC cells in chicken mince in real time and during experimental infection of it by Staphylococcus aureus 209P. In order to detect VBNC cells in chicken mince, total number of microbes, number of bacterial colonies (CFU), and the portion of living (dead) cells were determined in 1g of the product using a commercial set of fluorescent dyes. A second study was carried out after 5 h of incubation of tested samples at room temperature. In samples of minced meat on the 4th day after production, more than 99 % of all detected living cells were VBNC. After 5-hour incubation of the sample, the number of CFU/g increased by 22.5 times, but the portion of VBNC cells remained higher than 99 % of viable bacteria. During artificial infection of the same batch of mince with S. aureus in broth culture at the stage of logarithmic growth, the amount of VBNC cells for 0 hours was 97.33 %. After 5 hours their number increased to 99.99 %. Probably, in the introduced culture of Staphylococcus at the stage of active reproduction, formation of VBNC bacteria did not occur, which initially reduced their number in the sample. After 5-h incubation, transition of bacteria to VBNC state was accelerated, possibly due to unfavorable conditions for the cell population (changes in trophic substrate, temperature, pH, etc.). Experimental data confirm presence of VBNC bacteria in chicken products that don’t grow on traditional nutrient media and showing a false negative result in traditional microbiological expertise. Because of the biohazard of such dormant cells, it is advisable to provide regulated testing of foodstuffs for presence of VBNC cells.

жизнеспособныено некультивируемые клеткикуриный фаршдетекциябиоопасностьviable but nonculturable cellschicken mincedetectionbiohazardReferencesАбдуллаева, А. М., Блинкова, Л. П., & Пахомов, Ю. Д. (2019) Значение жизнеспособных некультивируемых бактерий для безопасности пищевых продуктов. Проблемы Ветеринарной Санитарии, Гигиены и Экологии, 2(30), с. 183-190.Abdullaeva, A. M., Blinkova, L. P., & Pakhomov, Yu. D. (2019). Significance of viable but nonculturable bacteria for safety of food products. Problemy Veterinarnoy, Sanitarii, Gigieny i Ekologii [Problems of Veterinary, Sanitation, Hygiene and Ecology] 2(30), 183-189. http://dx.doi.org/10.25725/vet.san.hyg.ecol.201902012Al-Qadiri, H. M., Lu, X., Al-Alami, N. I., & Rasco, B. A. (2011). Survival of Escherichia coli O157: H7 and Campylobacter jejuni in bottled purified drinking water under different storage conditions. Journal of Food Protection, 74(2), 254–260. https://doi.org/10.4315/0362-028X.JFP-10-368Al-Qadiri, H. M., Lu, X., Al-Alami, N. I., & Rasco, B. A. (2011). Survival of Escherichia coli O157: H7 and Campylobacter jejuni in bottled purified drinking water under different storage conditions. Journal of Food Protection, 74(2), 254–260. https://doi.org/10.4315/0362-028X.JFP-10-368Asakura, H., Makino, S.-I., Takagi, T., Kuri, A., Kurazono, T., Watarai, M., & Shirahata, T. (2002). Passage in mice causes a change in the ability of Salmonella enterica serovar Oranienburg to survive NaCl osmotic stress: resuscitation from the viable but non-culturable state. FEMS Microbiology Letters, 212(1), 87-93. https://doi.org/10.1111/j.1574-6968.2002.tb11249.xAsakura, H., Makino, S.-I., Takagi, T., Kuri, A., Kurazono, T., Watarai, M., & Shirahata, T. (2002). Passage in mice causes a change in the ability of Salmonella enterica serovar Oranienburg to survive NaCl osmotic stress: resuscitation from the viable but non-culturable state. FEMS Microbiology Letters, 212(1), 87-93. https://doi.org/10.1111/j.1574-6968.2002.tb11249.xAurass, P., Prager, R., & Flieger, A. (2011). EHEC/ EAEC O104: H4 strain linked with the 2011 German outbreak of haemolytic uremic syndrome enters into the viable but non-culturable state in response to various stresses and resuscitates upon stress relief. Environmental Microbiology, 13(12), 3139–3148. https://doi.org/10.1111/j.1462-2920.2011.02604.xAurass, P., Prager, R., & Flieger, A. (2011). EHEC/ EAEC O104: H4 strain linked with the 2011 German outbreak of haemolytic uremic syndrome enters into the viable but non-culturable state in response to various stresses and resuscitates upon stress relief. Environmental Microbiology, 13(12), 3139–3148. https://doi.org/10.1111/j.1462-2920.2011.02604.xAyrapetyan, M., & Oliver, J. D. (2016). The viable but non-culturable state and its relevance in food safety. Current Opinion in Food Science, 8, 127–133. https://doi.org/10.1016/j.cofs.2016.04.010Ayrapetyan, M., & Oliver, J. D. (2016). The viable but non-culturable state and its relevance in food safety. Current Opinion in Food Science, 8, 127–133. https://doi.org/10.1016/j.cofs.2016.04.010Barron, J. C., & Forsythe, S. J. (2007). Dry stress and survival time of Enterobacter sakazakii and other Enterobacteriaceae in dehydrated powdered infant formula. Journal of Food Protection, 70(9), 2111–2117.https://doi.org/10.4315/0362-028X-70.9.2111Barron, J. C., & Forsythe, S. J. (2007). Dry stress and survival time of Enterobacter sakazakii and other Enterobacteriaceae in dehydrated powdered infant formula. Journal of Food Protection, 70(9), 2111–2117.https://doi.org/10.4315/0362-028X-70.9.2111Bates, T. C., & Oliver, J. D. (2004). The viable but nonculturable state of Kanagawa positive and negative strains of Vibrio parahaemolyticus. Journal of Microbiology, 42(2), 74–79.Bates, T. C., & Oliver, J. D. (2004). The viable but nonculturable state of Kanagawa positive and negative strains of Vibrio parahaemolyticus. Journal of Microbiology, 42(2), 74–79.Blinkova, L., Martirosyan, D., Pakhomov, Yu., Dmitrieva, O., Vaughan, R., & Altshuler M. (2014). Nоnculturable forms of bacteria in lyophilized probiotic preparations. Functional Foods in Health and Disease, 4(2), 66-76.Blinkova, L., Martirosyan, D., Pakhomov, Yu., Dmitrieva, O., Vaughan, R., & Altshuler M. (2014). Nоnculturable forms of bacteria in lyophilized probiotic preparations. Functional Foods in Health and Disease, 4(2), 66-76.Capozzi, V., Di Toro, M. R., Grieco, F., Michelotti, V., Salma, M., Lamontanara, A., Russo, P., Orru, L., Alexandre, H., & Spano, G. (2016). Viable But Not Culturable (VBNC) state of Brettanomyces bruxellensis in wine: new insights on molecular basis of VBNC behaviour using a transcriptomic approach. Food Microbiology, 59, 196–204. https://doi.org/10.1016/j.fm.2016.06.007Capozzi, V., Di Toro, M. R., Grieco, F., Michelotti, V., Salma, M., Lamontanara, A., Russo, P., Orru, L., Alexandre, H., & Spano, G. (2016). Viable But Not Culturable (VBNC) state of Brettanomyces bruxellensis in wine: new insights on molecular basis of VBNC behaviour using a transcriptomic approach. Food Microbiology, 59, 196–204. https://doi.org/10.1016/j.fm.2016.06.007Chaisowwong, W., Kusumoto, A., Hashimoto, M., Harada, T., Maklon, K., & Kawamoto, K. (2012). Physiological characterization of Campylobacter jejuni under cold stresses conditions: its potential for public threat. Journal of Veterinary Medical Science, 74(1), 43–50. https://doi.org/10.1292/jvms.11-0305Chaisowwong, W., Kusumoto, A., Hashimoto, M., Harada, T., Maklon, K., & Kawamoto, K. (2012). Physiological characterization of Campylobacter jejuni under cold stresses conditions: its potential for public threat. Journal of Veterinary Medical Science, 74(1), 43–50. https://doi.org/10.1292/jvms.11-0305Cunningham, E., O’Byrne, C., & Oliver, J. D. (2009). Effect of weak acids on Listeria monocytogenes survival: evidence for a viable but nonculturable state in response to low pH. Food Control, 20(12), 1141–1144. https://doi.org/10.1016/j.foodcont.2009.03.005Cunningham, E., O’Byrne, C., & Oliver, J. D. (2009). Effect of weak acids on Listeria monocytogenes survival: evidence for a viable but nonculturable state in response to low pH. Food Control, 20(12), 1141–1144. https://doi.org/10.1016/j.foodcont.2009.03.005Ding, T., Suo, Y., Xiang, Q., Zhao, X., Chen, S., Ye, X., & Liu, D. (2017). Significance of viable but nonculturable Escherichia coli: induction, detection, and control. Journal of Microbiology and Biotechnology, 27(3), 417–428 https://doi.org/10.4014/jmb.1609.09063.Ding, T., Suo, Y., Xiang, Q., Zhao, X., Chen, S., Ye, X., & Liu, D. (2017). Significance of viable but nonculturable Escherichia coli: induction, detection, and control. Journal of Microbiology and Biotechnology, 27(3), 417–428 https://doi.org/10.4014/jmb.1609.09063.Dinu, L. D., & Bach, S. (2013). Detection of viable but non-culturable Escherichia coli O157: H7 from vegetable samples using quantitative PCR with propidium monoazide and immunological assays. Food Control, 31(2), 268-273.Dinu, L. D., & Bach, S. (2013). Detection of viable but non-culturable Escherichia coli O157: H7 from vegetable samples using quantitative PCR with propidium monoazide and immunological assays. Food Control, 31(2), 268-273.Divol B. & Lonvaud-Funel A. (2004). Evidence for viable but nonculturable yeasts in botrytis – affected wine. Journal of Applied Microbiology, 99(1), 85-93.Divol B. & Lonvaud-Funel A. (2004). Evidence for viable but nonculturable yeasts in botrytis – affected wine. Journal of Applied Microbiology, 99(1), 85-93.Fakruddin, M., Mannan, K. S., & Andrews, S. (2013). Viable but nonculturable bacteria: food safety and public health perspective. ISRN Microbiology, 2013:703813. https://doi.org/10.1155/2013/703813Fakruddin, M., Mannan, K. S., & Andrews, S. (2013). Viable but nonculturable bacteria: food safety and public health perspective. ISRN Microbiology, 2013:703813. https://doi.org/10.1155/2013/703813Gunasekera, T. S., Sørensen, A., Attfield, P. V., Sørensen, S. J., & Veal, D. A. (2002). Inducible gene expression by nonculturable bacteria in milk after pasteurization. Applied and Environmental Microbiology, 68(4), 1988–1993. https://doi.org/10.1128/AEM.68.4.1988-1993.2002Gunasekera, T. S., Sørensen, A., Attfield, P. V., Sørensen, S. J., & Veal, D. A. (2002). Inducible gene expression by nonculturable bacteria in milk after pasteurization. Applied and Environmental Microbiology, 68(4), 1988–1993. https://doi.org/10.1128/AEM.68.4.1988-1993.2002Hu, Y., & Coates, A. (2012). “Nonmultiplying bacteria are profoundly tolerant to antibiotics,” in Antibiotic Resistance, eds R. M. Anthony and A. Coates (Berlin: Springer Press), 99–119. https://doi.org/ 10.1007/978-3-642-28951-4_7Hu, Y., & Coates, A. (2012). “Nonmultiplying bacteria are profoundly tolerant to antibiotics,” in Antibiotic Resistance, eds R. M. Anthony and A. Coates (Berlin: Springer Press), 99–119. https://doi.org/10.1007/978-3-642-28951-4_7Kramer, B., & Muranyi, P. (2014). Effect of pulsed light on structural and physiological properties of Listeria innocua and Escherichia coli. Journal of Applied Microbiology, 116(3), 596–611. https://doi.org/10.1111/jam.12394Kramer, B., & Muranyi, P. (2014). Effect of pulsed light on structural and physiological properties of Listeria innocua and Escherichia coli. Journal of Applied Microbiology, 116(3), 596–611. https://doi.org/10.1111/jam.12394Li, L., Mendis, N., Trigui, H., Oliver, J. D., & Faucher, S. P. (2014). The importance of the viable but nonculturable state in human bacterial pathogens. Frontiers in Microbiology, 5, 258. https://doi.org/10.3389/fmicb.2014.00258Li, L., Mendis, N., Trigui, H., Oliver, J. D., & Faucher, S. P. (2014). The importance of the viable but nonculturable state in human bacterial pathogens. Frontiers in Microbiology, 5, 258. https://doi.org/10.3389/fmicb.2014.00258Liu, J., Li, L., Li, B., Peters, B. M., Deng, Y., Xu, Z. & Shirtliff, M. E. (2017). First study on the formation and resuscitation of viable but nonculturable state and beer spoilage capability of Lactobacillus lindneri. Microbial Pathogenesis, 107, 219-224. https://doi.org/ 10.1016/j.micpath.2017.03.043Liu, J., Li, L., Li, B., Peters, B. M., Deng, Y., Xu, Z. & Shirtliff, M. E. (2017). First study on the formation and resuscitation of viable but nonculturable state and beer spoilage capability of Lactobacillus lindneri. Microbial Pathogenesis, 107, 219-224. https://doi.org/ 10.1016/j.micpath.2017.03.043Makino, S.-I., Kii, T., Asakura, H., Shirahata, T., Ikeda, T., Takeshi, K., & Itoh, K. (2000). Does Enterohemorrhagic Escherichia coli O157: H7 enter the viable but nonculturable state in salted salmon roe? Applied and Environmental Microbiology, 66(12), 5536–5539. https://doi.org/10.1128/AEM.66.12.5536-5539.2000Makino, S.-I., Kii, T., Asakura, H., Shirahata, T., Ikeda, T., Takeshi, K., & Itoh, K. (2000). Does Enterohemorrhagic Escherichia coli O157: H7 enter the viable but nonculturable state in salted salmon roe? Applied and Environmental Microbiology, 66(12), 5536–5539. https://doi.org/10.1128/AEM.66.12.5536-5539.2000Masmoudi, S., Denis, M., & Maalej, S. (2010). Inactivation of the gene katA or sodA affects the transient entry into the viable but nonculturable response of Staphylococcus aureus in natural seawater at low temperature. Marine Pollution Bulletin, 60(12), 2209–2214. https://doi.org/10.1016/j.marpolbul.2010.08.017Masmoudi, S., Denis, M., & Maalej, S. (2010). Inactivation of the gene katA or sodA affects the transient entry into the viable but nonculturable response of Staphylococcus aureus in natural seawater at low temperature. Marine Pollution Bulletin, 60(12), 2209–2214. https://doi.org/10.1016/j.marpolbul.2010.08.017Nicolò, M. S., Gioffrè, A., Carnazza, S., Platania, G., Silvestro, I. D., & Guglielmino, S. P. (2010). Viable but nonculturable state of foodborne pathogens in grapefruit juice: a study of laboratory. Foodborne Pathogens and Disease, 8(1), 11–17. https://doi.org/10.1089/fpd.2009.0491Nicolò, M. S., Gioffrè, A., Carnazza, S., Platania, G., Silvestro, I. D., & Guglielmino, S. P. (2010). Viable but nonculturable state of foodborne pathogens in grapefruit juice: a study of laboratory. Foodborne Pathogens and Disease, 8(1), 11–17. https://doi.org/10.1089/fpd.2009.0491Nicolò, M. S., & Guglielmino, S. P. P. (2012). “Viable but nonculturable bacteria in food,” in Public Health–Methodology, Environmental and Systems Issues, ed. J. Maddock (Rjeka: InTech), 189–216. https://doi.org/10.5772/38118Nicolò, M. S., & Guglielmino, S. P. P. (2012). “Viable but nonculturable bacteria in food,” in Public Health–Methodology, Environmental and Systems Issues, ed. J. Maddock (Rjeka: InTech), 189–216. https://doi.org/10.5772/38118Nyström, T. (2003). Nonculturable bacteria: programmed survival forms or cells at death’s door? Bioessays, 25(3), 204–211. https://doi.org/10.1002/bies.10233Nyström, T. (2003). Nonculturable bacteria: programmed survival forms or cells at death’s door? Bioessays, 25(3), 204–211. https://doi.org/10.1002/bies.10233Oh, E., McMullen, L., & Jeon, B. (2015). Impact of oxidative stress defense on bacterial survival and morphological change in Campylobacter jejuni under aerobic conditions. Frontiers in Microbiology, 6, 295. https://doi.org/10.3389/fmicb.2015.00295Oh, E., McMullen, L., & Jeon, B. (2015). Impact of oxidative stress defense on bacterial survival and morphological change in Campylobacter jejuni under aerobic conditions. Frontiers in Microbiology, 6, 295. https://doi.org/10.3389/fmicb.2015.00295Oliver, J. D. (2010). Recent findings on the viable but nonculturable state in pathogenic bacteria. FEMS Microbiology Reviews, 34(4), 415-425. https://doi.org/10.1111/j.1574-6976.2009.00200.xOliver, J. D. (2010). Recent findings on the viable but nonculturable state in pathogenic bacteria. FEMS Microbiology Reviews, 34(4), 415-425. https://doi.org/10.1111/j.1574-6976.2009.00200.xPeneau, S., Chassaing, D., & Carpentier, B. (2007). First evidence of division and accumulation of viable but nonculturable Pseudomonas fluorescens cells on surfaces subjected to conditions encountered at meat processing premises. Applied and Environmental Microbiology, 73(9), 2839–2846. https://doi.org/10.1128/AEM.02267-06Peneau, S., Chassaing, D., & Carpentier, B. (2007). First evidence of division and accumulation of viable but nonculturable Pseudomonas fluorescens cells on surfaces subjected to conditions encountered at meat processing premises. Applied and Environmental Microbiology, 73(9), 2839–2846. https://doi.org/10.1128/AEM.02267-06Pinto D., Santos M. A., & Chambel L. (2015). Thirty years of viable but nonculturable state research unsolved molecular mechanisms. Critical Reviews in Microbiology, 41(1), 61-76.Pinto D., Santos M. A., & Chambel L. (2015). Thirty years of viable but nonculturable state research unsolved molecular mechanisms. Critical Reviews in Microbiology, 41(1), 61-76.Ramamurthy, T., Ghosh, A., Pazhani, G. P., & Shinoda, S. (2014). Current Perspectives on Viable but Non- Culturable (VBNC) Pathogenic Bacteria. Frontiers in Public Health, 2, 103. https://doi.org/10.3389/fpubh.2014.00103Ramamurthy, T., Ghosh, A., Pazhani, G. P., & Shinoda, S. (2014). Current Perspectives on Viable but Non- Culturable (VBNC) Pathogenic Bacteria. Frontiers in Public Health, 2, 103. https://doi.org/10.3389/fpubh.2014.00103Rao, N. V., Shashidhar, R., & Bandekar, J. R. (2014). Induction, resuscitation and quantitative realtime polymerase chain reaction analyses of viable but nonculturable Vibrio vulnificus in artificial sea water. World Journal of Microbiology and Biotechnology, 30, 2205–2212. https://doi.org/10.1007/s11274-014-1640-1Rao, N. V., Shashidhar, R., & Bandekar, J. R. (2014). Induction, resuscitation and quantitative realtime polymerase chain reaction analyses of viable but nonculturable Vibrio vulnificus in artificial sea water. World Journal of Microbiology and Biotechnology, 30, 2205–2212. https://doi.org/10.1007/s11274-014-1640-1Rowan, N. J. (2004). Viable but non-culturable forms of food and waterborne bacteria: quo vadis? Trends in Food Science & Technology, 15(9) 462–467. https://doi.org/10.1016/j.tifs.2004.02.009Rowan, N. J. (2004). Viable but non-culturable forms of food and waterborne bacteria: quo vadis? Trends in Food Science & Technology, 15(9) 462–467. https://doi.org/10.1016/j.tifs.2004.02.009Rowan, N. J., Valdramidis, V. P., & Gomez-Lopez, V. M. (2015). A review of quantitative methods to describe efficacy of pulsed light generated inactivation data that embraces the occurrence of viable but non culturable state microorganisms. Trends in Food Science & Technology, 44(1), 79–92. https://doi.org/10.1016/j.tifs.2015.03.006Rowan, N. J., Valdramidis, V. P., & Gomez-Lopez, V. M. (2015). A review of quantitative methods to describe efficacy of pulsed light generated inactivation data that embraces the occurrence of viable but non culturable state microorganisms. Trends in Food Science & Technology, 44(1), 79–92. https://doi.org/10.1016/j.tifs.2015.03.006Schottroff, F., Fröhling, A., Zunabovic-Pichler, M., Krottenthaler, A., Shlüter, O., & Jäger, H. (2018). Sublethal injury and viable but nonculturable (VBNC) state in microorganisms during preservation of food and biological materials by non-thermal processes. Frontiers in Microbiology, 9, article 2773, p. 1-19. https://doi.org/10.3389/fmicb.2018.02773.Schottroff, F., Fröhling, A., Zunabovic-Pichler, M., Krottenthaler, A., Shlüter, O., & Jäger, H. (2018). Sublethal injury and viable but nonculturable (VBNC) state in microorganisms during preservation of food and biological materials by non-thermal processes. Frontiers in Microbiology, 9, article 2773, p. 1-19. https://doi.org/10.3389/fmicb.2018.02773.Serpaggi, V., Remize, F., & Recorbet, G. (2012). Characterization of the “viable but nonculturable” (VBNC) state in the wine spoilage yeast Brettanomyces. Food Microbiology, 30(2), 440-447.Serpaggi, V., Remize, F., & Recorbet, G. (2012). Characterization of the “viable but nonculturable” (VBNC) state in the wine spoilage yeast Brettanomyces. Food Microbiology, 30(2), 440-447.Zhang, S., Ye, C., Lin, H., Lv, L., & Yu, X. (2015). UV disinfection induces a VBNC state in Escherichia coli and Pseudomonas aeruginosa. Environmental Science & Technology, 49(3), 1721–1728. https://doi.org/ 10.1021/es505211eZhang, S., Ye, C., Lin, H., Lv, L., & Yu, X. (2015). UV disinfection induces a VBNC state in Escherichia coli and Pseudomonas aeruginosa. Environmental Science & Technology, 49(3), 1721–1728. https://doi.org/ 10.1021/es505211eZhao, X., Zhong, J., Wei, C. Lin C.-W., & Ding T. (2017). Current perspectives on viable but nonculturable state in foodborne pathogens. Frontiers in Microbiology, 8, 1-32. https://doi.org/10.3389/fmicb.2017.00580Zhao, X., Zhong, J., Wei, C. Lin C.-W., & Ding T. (2017). Current perspectives on viable but nonculturable state in foodborne pathogens. Frontiers in Microbiology, 8, 1-32. https://doi.org/10.3389/fmicb.2017.00580Zhao, X., Wei, C., Zhong, J., & Jin, S. (2016). Research advance in rapid detection of foodborne Staphylococcus aureus. Biotechnology & Biotechnological Equipment, 30(5), 1–7. https://doi.org/ 10.1080/13102818.2016.1209433Zhao, X., Wei, C., Zhong, J., & Jin, S. (2016). Research advance in rapid detection of foodborne Staphylococcus aureus. Biotechnology & Biotechnological Equipment, 30(5), 1–7. https://doi.org/ 10.1080/13102818.2016.1209433Ziprin, R. L., Droleskey, R. E., Hume, M. E., & Harvey, R. B. (2003). Failure of viable nonculturable Campylobacter jejuni to colonize the cecum of newly hatched leghorn chicks. Avian Diseases, 47(3), 753–758. https://doi.org/10.1637/7015Ziprin, R. L., Droleskey, R. E., Hume, M. E., & Harvey, R. B. (2003). Failure of viable nonculturable Campylobacter jejuni to colonize the cecum of newly hatched leghorn chicks. Avian Diseases, 47(3), 753–758. https://doi.org/10.1637/7015

The authors declare that there are no conflicts of interest present.