Study of tolerance of enterobacteria to chlorine-based biocides in experimental models using chromogenic indicator tests

View or download the full article: 

N.R. Efimochkina, I.B. Bykova, Yu.V. Korotkevich, Yu.M. Markova, L.P. Minaeva, S.A. Sheveleva


FSBI "Institute of Nutrition", Russian Federation, Moscow, 2/14 Ustinsky Passage, 109240


The species-specific composition of microbial contaminants of vegetable raw materials and equipment used in the production of biotechnological products and beverages fermentation are studied. 85 enterobacteria strains was isolated and investigated, 46 strains of the genera Enterobacter, Pantoea, Citrobacter, Serratia, Escherichia, Cronobacter was identified to the species level; the most frequently detected bacteria of the genera Enterobacter and Pantoea (about 50 %). For the first time developed and tested chromogenic in vitro model based that allows to quantify the degree of inhibition of gram-negative microflora under the influence of antimicrobial agents depending on the concentrations of biocides and density of bacterial populations. A comparative analysis of the tolerance of Enterobacteriaceae strains from different biotopes was conducted. Sensitivity to the treatment of chlorine-containing biocides in 26 strains of enterobacteria from plant material and 9 strains of Escherichia coli from the intestine of male rats of Wistar line was tested. Enterobacteria from vegetable raw materials and swabs were more resistant to antimicrobial action of chlorine, than the representatives of the populations of the normal intestinal microbiota. It is established that the active chlorine concentration of 50–100 mg/dm 3 , the most commonly used in the processing of vegetable raw materials, is not effective for Enterobacteriaceae, if the density of the microbial population is 10 5–7 cells/cm 3 and above. At an initial level of contamination with Enterobacteriaceae not more than 10 3 cells/cm 3 processing solutions with a concentration of active chlorine of 75 to 100 mg/dm 3 can provide effective disinfection of raw materials, equipment, or inventory. Experimental chromogenic in vitro model proposed to assess the impact of chlorine-based biocides on the degree of the enterobacteria inhibition, can be used to justify the selection and doses of antimicrobial agents, effective against other groups of microbial contaminants. This will optimize the use of modes of decontamination of raw materials and sanitizing equipment in the food industry.

enterobacteria, chromogenic model in vitro, vegetable raw materials, chlorine-based biocides, tolerance
Efimochkina N.R., Bykova I.B., Korotkevich Yu.V., Markova Yu.M., Minaeva L.P. , Sheveleva S.A. Study of tolerance of enterobacteria to chlorine-based biocides in experimental models using chromogenic indicator tests. Health Risk Analysis, 2015, no. 3, pp. 73-82. DOI: 10.21668/health.risk/2015.3.11.eng
  1. Efimochkina N.R. Mikrobiologija pishhevyh produktov i sovremennye metody detekcii patogenov [Food microbiology and modern methods of detection of foodborne pathogens]. Мoscow: Publishing house of the Russian Academy of Medical Sciences, 2013. 517 p.
  2. Efimochkina N.R., Bykova I.B., Batisheva S.Yu., Minaeva L.P., Markova Yu.M., KorotkevichYu.V., Shi-lov G.Yu., ShevelevaS.A. Izuchenie osobennostej mikrobnoj kontaminacii svezhih ovoshhej i listovyh salatov pro-myshlennogo izgotovlenija [Study of microbial contamination of processed fresh vegetables and lettuce]. Voprosi pitanya, 2014, no. 5, pp. 33–42 (In Russan).
  3. SanPiN «Pit'evaja voda. Gigienicheskie trebovanija k kachestvu vody centralizovannyh sis-tem pit'evogo vodosnabzhenija. Kontrol' kachestva. Gigienicheskie trebovanija k obespecheniju bezopasnosti sis-tem gorjachego vodosnabzhenija. Sanitarno-jepidemiologicheskie pravila i normativy» [SanPin «Drinking water. Hygienic requirements to water quality of centralized drinking water supply systems. Quality con-trol. Hygienic requirements for safety of hot water systems. Sanitary-epidemiological rules and regulations»]. Mos-cow: Rospotrebnadzor, 2001. 112 р. (In Russan).
  4. Mohyla P., Bilgili S. F., Oyarzabal O. A. et al. Application of Acidified Sodium Chlorite in the Drinking Water to Control Salmonella serotype Typhimurium and Campylobacter jejuni in Commercial Broilers. J. Appl. Poult Res., 2007, vol. 16, no. 1, pp. 45–51. doi: 10.1093/japr/16.1.45.
  5. Christian M.S., York R.G., Hoberman, A.M. et al. Biodisposition of dibromoacetic acid (DBA) and bromo-dichloromethane (BDCM) administered to rats and rabbits in drinking water during range-finding reproduction and developmental toxicity studies. International Journal of Toxicology, 2001, vol. 20, pp. 239–253.
  6. Hicks S.J., Rowbury R.J. Resistance of attached Escherichia coli to acrylic acid and its significance for the survival of plasmid-bearing organisms in water. Ann. Inst. Pasteur, 1987, vol. 138, pp. 359–369.
  7. International Life Sciences Institute. An Evaluation of EPA’s Proposed Guidelines for Carcinogen Risk As-sessment Using Chloroform and Dichloroacetate as Case Studies. Report of an Expert Panel, ILSI HESI, Washing-ton, DC, November 1997. 240 р.
  8. Christian M.S., York R.G., Hoberman A.M. et al. Oral (drinking water) developmental toxicity studies of bromodichloromethane (BDCM) in rats and rabbits. International Journal of Toxicology, 2001, vol. 20, pp. 225–237.
  9. Christian M.S., York R.G., Hoberman A.M. et al. Oral (drinking water) two-generation reproductive toxicity study of bromodichloromethane (BDCM) in rats. International Journal of Toxicology, 2002, vol. 21, pp. 115–146.
  10. Oyarzabal O.A. Reduction of Campylobacter spp. by commercial antimicrobials applied during the processing of broiler chickens: a review from the United States perspective. J. Food Prot., 2005, vol. 68, pp. 1752–1760.
  11. Scientific Opinion of the Panel on Biological Hazards on a request from DG SANCO on the assessment of the possible effect of the four antimicrobial treatment substances on the emergence of antimicrobial resistance. The EFSA Journal, 2008, vol. 659, pp. 1–26.
  12. Whiting G.C., Rowbury R.J. Increased resistance of Escherichia coli to acrylic acid and to copper ions af-ter cold-shock. Letts. Appl. Microbiol., 1995, vol. 20, pp. 240–242.
  13. Yang, H., Li, Y. Johnson, M.G. Survival and death of Salmonella typhimurium and Campylobacter jejuni in processing water and on chicken skin during poultry scalding and chilling. J. Food Prot., 2001, vol. 64, pp. 770–776.

You are here