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

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

Preparation of goat and rabbit anti-camel immunoglobulin G whole molecule labeled with horseradish peroxidase

Aim: As the labeled anti-camel immunoglobulins (Igs) with enzymes for enzyme-linked immunosorbent assay (ELISA) are unavailable in the Egyptian market, the present investigation was directed for developing local labeled anti-camel IgG with horseradish peroxidase (HRP) to save hard curacy. Materials and Methods: For purification of camel IgG whole molecule, camel sera was preliminary precipitated with 50% saturated ammonium sulfate and dialyzed against 15 mM phosphate-buffered saline pH 7.2 then concentrated. This preparation was further purified by protein A sepharose affinity column chromatography. The purity of the eluted camel IgG was tested by sodium dodecyl sulfate polyacrylamide gel electrophoresi. Anti-camel IgG was prepared by immunization of goats and rabbits separately, with purified camel IgG. The anti-camel IgG was purified by protein A sepharose affinity column chromatography. Whole molecule anti-camel IgG was conjugated with HRP using glutraldehyde based assay. Sensitivity and specificity of prepared conjugated secondary antibodies were detected using positive and negative camel serum samples reacted with different antigens in ELISA, respectively. The potency of prepared conjugated antibodies was evaluated compared with protein A HRP. The stability of the conjugate at −20°C during 1 year was assessed by ELISA. Results: The electrophoretic profile of camel IgG showed four bands of molecular weight 63, 52, 40 and 33 kDa. The recorded sensitivity and specificity of the product are 100%. Its potency is also 100% compared to 58-75% of commercial protein A HRP. The conjugates are stable for 1 year at −20°C as proved by ELISA. Conclusion: Collectively, this study introduces goat and rabbit anti-camel IgG whole molecules with simple, inexpensive method, with 100% sensitivity, 100% specificity and stability up to 1 year at −20°C. The important facet of the current study is saving hard curacy. Future investigations are necessary for preparation of IgG subclasses

Isolation of antimicrobial producing Actinobacteria from soil samples

Emergence of multidrug resistant bacteria has made the search for novel bioactive compounds from natural and unexplored habitats a necessity. Actinobacteria have important bioactive substances. The present study investigated antimicrobial activity of Actinobacteria isolated from soil samples of Egypt. One hundred samples were collected from agricultural farming soil of different governorates. Twelve isolates have produced activity against the tested microorganisms (S. aureus, Bacillus cereus, E. coli, K. pneumoniae, P. aeruginosa, S. Typhi, C. albicans, A. niger and A. flavus). By VITEK 2 system version: 07.01 the 12 isolates were identified as Kocuria kristinae, Kocuria rosea, Streptomyces griseus, Streptomyces flaveolus and Actinobacteria. Using ethyl acetate extraction method the isolates culture’s supernatants were tested by diffusion method against indicator microorganisms. These results indicate that Actinobacteria isolated from Egypt farms could be sources of antimicrobial bioactive substances