Evaluation of the efficacy of commonly used disinfectants against isolated chlorine-resistant strains from drinking water used in Egyptian cattle farms

Abstract

Background and aim: Drinking water of poor microbiological quality contains high percentages of microbes causing outbreaks of mainly coliform-related diseases. These microbes could be controlled by many hygienic standards including disinfection, but disinfectants misuse causes the developing of disinfectant-resistant strains. The present study aimed to investigate drinking water bacterial profile, determine chlorine-resistant strains, and statistically correlate that with the used disinfectant and disinfection process variables. In vitro evaluation of the bactericidal effect of the most commonly used disinfectants in cattle operations against the isolated chlorine-resistant strains and detection of qacE resistance gene in the isolated chlorine-resistant Escherichia coli strains in some cattle farms suffering coliform and non-coliform related disease around Egypt.

Materials and methods: A structured questionnaire is used to survey a convenience sample of 132 Egyptian cattle beef and dairy farms suffering emerged epidemics to identify commonly used disinfection process, disinfectant types, disinfectants frequency, and rate of use. One hundred and thirty-two water samples were collected for microbiological analysis to obtain water bacterial profile and testing resistance to chlorine. Statistical analysis was performed to identify the level of association between microbial profile and presence of chlorine-resistant strains in each farm with used disinfection, disinfectant types, and rate of use in these farms.

Results: A wide range of disinfectant types used for variable purposes inside cattle farms with a different frequency of use and the highest percent of farms 25.8% use 4-5 types of disinfectants, followed by 25% of farms use two types, then 18.9% use three types. Microbial profile of water samples revealed isolation of E. coli, Streptococcus faecalis, Pseudomonas aeruginosa, Klebsiella spp., Proteus spp., Salmonella spp., Enterobacter spp., Citrobacter spp., Shigella flexneri, Serratia marcescens, and Yersinia enterocolitica in percent (98.5, 97.7, 97.7, 76.5, 66.7, 36.4, 78.8, 74.2, 30.3, 29.5, and 14.4% of cattle farms, respectively), from which five E. coli, four Salmonella, four Pseudomonas, two Klebsiella, and four Streptococcus strains expressed chlorine resistance. Statistical analysis showed weak to moderate correlation (rho 0.15-0.46) between bacterial profile strains count and presence of resistant strains with different farm disinfection, disinfectant types, and rate of use. Experimental evaluation of the bactericidal effect of the eight selected disinfectants on the chlorine-resistant isolated strains revealed that peroxymonosulfate killed 19/19 isolated strains/15 min contact time, and quaternary ammonium compounds killed only 3/19 strains/15 min contact time. The qacE resistance gene was detected in 3/4 isolated chlorine-resistant E. coli strains.

Conclusion: Drinking water microbial profile strains and resistance to disinfectants are widely varied in cattle farms, and this variance depends on critical factors among which the disinfection process types used disinfectant types and frequency of disinfectants use or change.

Keywords: cattle farms; disinfectant resistance; drinking water; microbial profile; resistance genes.

Figure-1

Spearman’s rho correlation coefficient between microbial profile count and presence of resistant strain, with particular disinfectant type (T) and disinfectant frequency of use or change (F).

Figure-2

The presence of qacE gene among four E. coli chlorine-resistant strains. Lane M=100 bp ladder, lane 1=positive control, lane 2=negative control, lane E1=sample code O1 had negative result, lane E2=sample code O2 had positive result, lane E3=sample code O5 had positive result, and lane E4=sample code O4 had positive result.