Efficacy evaluation of hydrogen peroxide disinfectant based zinc oxide nanoparticles against diarrhea causing Escherichia coli in ruminant animals and broiler chickens.

Different strains of Escherichia coli that exhibit genetic characteristics linked to diarrhea pose a major threat to both human and animal health. The purpose of this study was to determine the prevalence of pathogenic Escherichia coli (E. coli), the genetic linkages and routes of transmission between E. coli isolates from different animal species. The efficiency of disinfectants such as hydrogen peroxide (H2O2), Virkon®S, TH4+, nano zinc oxide (ZnO NPs), and H2O2-based zinc oxide nanoparticles (H2O2/ZnO NPs) against isolated strains of E. coli was evaluated. Using 100 fecal samples from different diarrheal species (cow n = 30, sheep n = 40, and broiler chicken n = 30) for E. coli isolation and identification using the entero-bacterial repetitive intergenic consensus (ERIC-PCR) fingerprinting technique. The E. coli properties isolated from several diarrheal species were examined for their pathogenicity in vitro. Scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HR-TEM), Fourier-transform infrared spectrum (FT-IR), X-ray diffraction (XRD), zeta potential, and particle size distribution were used for the synthesis and characterization of ZnO NPs and H2O2/ZnO NPs. The broth macro-dilution method was used to assess the effectiveness of disinfectants and disinfectant-based nanoparticles against E. coli strains. Regarding the results, the hemolytic activity and Congo red binding assays of pathogenic E. coli isolates were 55.3 and 44.7%, respectively. Eleven virulent E. coli isolates were typed into five ERIC-types (A1, A2, B1, B2, and B3) using the ERIC-PCR method. These types clustered into two main clusters (A and B) with 75% similarity. In conclusion, there was 90% similarity between the sheep samples' ERIC types A1 and A2. On the other hand, 89% of the ERIC types B1, B2, and B3 of cows and poultry samples were comparable. The H2O2/ZnO NPs composite exhibits potential antibacterial action against E. coli isolates at 0.04 mg/ml after 120 min of exposure.

The Efficacy of Bacteriocins Against Biofilm-Producing Bacteria Causing Bovine Clinical Mastitis in Dairy Farms: A New Strategy.

Using an alternative bio-product is one of the most promising ways to control bovine mastitis and avoid new intra-mammary infections. The aims of this study were to ascertain the prevalence of biofilm-forming bacteria responsible for causing clinical mastitis in dairy herds and to assess the effectiveness of bacteriocins, produced by Bacillus subtilis, in controlling the growth of these bacteria in the milk of animals. A total of 150 milk samples were collected from cows and buffalos suffering from mastitis and the etiological agents were isolated and identified by the VITEK-2-COMPACT-SYSTEM®. Additionally, the capability of the bacterial isolates to produce biofilms was determined. RT-PCR was used to detect enterotoxin-producing genes (sed and seb), resistance genes (mecA and blaZ), and biofilm-associated genes (icaA and fnbA) in the isolated bacteria. The susceptibility patterns of the bacterial isolates to bacteriocins were assessed using an agar well-diffusion assay. S. aureus was significantly more capable of producing biofilms than coagulase-negative Staphylococcus isolates. S. ubris was the strongest biofilm producer among the Streptococcus species. The sensitivity profiles of the Staphylococcus spp. (S. aureus and coagulase-negative Staphylococcus) and their biofilm producers to bacteriocins were significantly higher (100% and 90%, respectively) at the same concentration. Bacteriocins had a lethal effect on Staphylococci, Streptococci, and biofilm development at a dose of 250 µg/mL. In dairy farms, bacteriocins are a viable alternative treatment for the prevention and control of bovine clinical mastitis.

An alternative approach for controlling bacterial pathogens in liquid and solid poultry waste using Calcium hypochlorite Ca(OCl)2 disinfectant-based silver nanoparticles.

Inappropriate handling of poultry waste from the beginning to the end of the production cycle could lead to health and environmental hazards. The purpose of this study was to assess the current state of poultry waste management practices as well as to evaluate the efficacy of disinfectants (VIRKON S, Quaternary ammonium compound (QAC), Calcium hypochlorite [Ca(OCl)2], and nanomaterials (nano-silver particles (Ag NPs), and Ca(OCl)2-Ag NPs composite) on pathogenic bacteria for use in the disinfection of waste collection areas within poultry operation systems. Two hundred and ten samples were gathered from variant waste types for isolation and identification of pathogenic bacteria. Then, the efficacy of some disinfectants against fifty strains of isolated bacteria was evaluated using a broth micro-dilution assay. Results showed the most predominant bacterial isolates from wastes were E. coli (33.69%), Salmonella spp. (26.09%), followed by K. pneumonae (15.22%) and L. monocytogenes (14.13%). Ca(OCl)2-Ag NPs had a microbial lethal effect against all pathogenic bacteria (100%) that were isolated from liquid and solid waste. In conclusion, poultry manure waste is collected and dumped on the agricultural land around those chicken farms without any treatment. The Ca(OCl)2-Ag NPs composite was lethal to all pathogenic microbes isolated from waste and their collected areas at 1.0 mg/L concentration.

Efficiency evaluation of some novel disinfectants and anti-bacterial nanocomposite on zoonotic bacterial pathogens in commercial Mallard duck pens for efficient control.

OBJECTIVE: This work aimed to detect the frequency of pathogenic bacteria of zoonotic importance in ducks' dropping, their surrounding environment, and farmworkers in contact with them. Furthermore, the susceptibility pattern of isolated bacteria to antimicrobial drugs and the efficiency of disinfectants (CID 20, Durak® plus, and hydrogen peroxide (H2O2), nano zinc oxide (ZnO NPs), and hydrogen peroxide loaded nano zinc oxide (H2O2/ZnO NPs) composites against isolated bacteria were evaluated. MATERIALS AND METHODS: A total of 271 samples were collected from duck pens, including 35 fecal droppings, 200 environmental samples, and 36 from the hands of pen workers for isolation and identification of bacterial strains using standard microbiological procedures. After that, the antibiotic sensitivity testing of 40 bacterial isolates was carried out using disk diffusion assay. ZnO NPs and H2O2/ZnO NPs were characterized using Fourier-transform infrared spectrum and high-resolution transmission electron microscopy. The efficacy of disinfectants and nanocomposites was evaluated against enteropathogenic bacteria using the broth macro-dilution method. RESULTS: The results showed that the overall prevalence of pathogenic bacteria in duck pens was 62.73. The highest isolation rate was detected in duck fecal droppings (100%), while Escherichia coli was found to be the most isolated pathogen (56.47%), followed by Pseudomonas aeruginosa (21.8%), Proteus mirabilis (15.29), and Salmonella species (6.47%). Multidrug resistance (MDR) was detected in the majority of bacterial isolates. The efficiency of CID 20 and Durak® plus disinfectants against all bacterial isolates was highly susceptible (100%) after 120 min of exposure time compared to the effectiveness of H2O2 on enteropathogenic bacteria which did not exceeded 60% at 5% concentration. Meanwhile, the sensitivity of Salmonella spp. to Durak® plus did not exceeded 80%. CONCLUSION: The duck fecal droppings are the primary source of bacterial isolates. MDR isolates were susceptible to both CID 20 and Durak® plus disinfectants after 120 min of exposure time at a concentration of 1:100 ml. Besides, H2O2/ZnO NPs composite proved its lethal effect against all testing strains at 0.02 mg/ml after 120 min of exposure. Strict biosecurity guidelines are required to mitigate and prevent the transmission of potentially zoonotic pathogens through the farm environment and/or duck droppings.

Exploitation of new approach to control of environmental pathogenic bacteria causing bovine clinical mastitis using novel anti-biofilm nanocomposite.

New approaches are required for prevention and control of biofilm-producing bacteria and consequently mitigating the health problems of bovine clinical mastitis. This work designed to determine prevalence rates of biofilm-producing bacteria that causing bovine clinical mastitis and evaluate the anti-biofilm effectiveness of novel nanocomposite of zinc-aluminum layered double hydroxide intercalated with gallic acid (GA) as chelating agent (Zn-Al LDH/GA) on the prevention and control of environmental pathogenic bacteria; Escherichia coli (E. coli), Klebsiella pneumoniae (K. pneumoniae), Staphylococcus aureus (S. aureus), and Coagulase-negative staphylococci (CNS), besides Listeria monocytogenes (L. monocytogenes) and assess the ability to use as an antimicrobial agent, and/or sanitizer for milking equipment. All samples (n = 230) involved clinical mastitis cow's milk (n = 50) beside environmental samples (n = 180) were collected then examined for isolation and identification of bacterial pathogens. Zn-Al LDH/GA nanocomposite was synthesized using co-precipitation method, then characterized by Fourier-transform infrared spectroscopy (FT-IR); X-ray diffraction (XRD); field emission scanning electron microscopy (FESEM); high-resolution transmission electron microscopy (HRTEM); thermogravimetric analysis (TGA); differential thermal analysis (DTA); zeta potential; DLS analysis; and Brunauer, Emmett, and Teller (BET) surface area. The anti-biofilm activity of nanocomposite against mastitis-causing bacteria was detected using the broth micro-dilution and disc-diffusion assay. Results, the minimum concentration of Zn-Al LDH/GA that inhibited the growth of gram-positive and negative bacteria, were 312-625 and 5000 µg/mL, respectively. The LD50 of Zn-Al LDH/GA was determined in mice at 1983.3 mg/kg b.wt. As a conclusion, Zn-Al LDH/GA nanocomposite proved its efficiency as an antimicrobial agent and/or sanitizer used for cleaning of milking equipment, due to it could inhibit the growth and multiplication of potentially pathogenic bacteria that causing clinical mastitis and its formation of biofilm on the milking equipment. Zn-Al LDH/GA was found to use under varying pH conditions compared with other commercial sanitizer used besides the formation of nanocomposite increases the material stability.

Acaricidal efficacy of deltamethrin-zinc oxide nanocomposite on Rhipicephalus (Boophilus) annulatus tick.

In the veterinary field, there is a great concern about the issue of acaricides resistance in cattle ticks. A comparative study was designed to evaluate the in vitro acaricidal efficacy of imidacloprid nano-emulsion, zinc oxide nanoparticles (ZnO NPs), silver nanoparticles (Ag NPs), deltamethrin, deltamethrin-ZnO NPs, and deltamethrin-Ag NPs. In addition, the novel formulations of deltamethrin (deltamethrin-ZnO NPs and deltamethrin-Ag NPs) against Rhipicephalus (Boophilus) annulatus were also evaluated by in vivo animal efficacy trials. Nanoparticles and their loaded forms, deltamethrin-Ag NPs and deltamethrin-ZnO NPs, were characterized by Transmission Electron Microscope (TEM) and Fourier-Transform Infrared Spectrum (FTIR). The adulticidal activity of deltamethrin-ZnO NPs at different concentrations; 2 mL/L, 1 mL/L, 0.5 mL/L and 0.25 mL/L induced a significant (P ≤ 0.05) lethal effect on adult ticks compared to deltamethrin-Ag NPs at the same concentrations. The larvicidal efficacy of deltamethrin-ZnO NPs resulted in a complete larval mortality within 24 h of exposure, while deltamethrin and deltamethrin-Ag NPs exhibited 100% immobility of larvae 48 h post-exposure. Furthermore, the in vivo experiments showed a considerable reduction in the tick's survival after using deltamethrin-ZnO NPs. There was no significant effect of different treatments on liver or kidney function tests at pre- and post-treatment of animals. In conclusion, deltamethrin-ZnO nanocomposite was the most effective adulticide and larvicide against R. (B.) annulatus. To the best of our knowledge, this is the first report for using deltamethrin-ZnO NPs as an acaricide.

Resistance of bacterial pathogens to calcium hypochlorite disinfectant and evaluation of the usability of treated filter paper impregnated with nanosilver composite for drinking water purification.

OBJECTIVES: Innovative techniques are urgently required to remove pathogenic bacterial contamination of drinking water. This study aimed to evaluate the biocidal activity of calcium hypochlorite [Ca(OCl)2], silver nanoparticles (AgNPs) and Ca(OCl)2/AgNPs composite against bacteria isolated from drinking water supplies (tap and hand pump water). A field trial was subsequently performed to evaluate the efficacy of a biocidal filter paper containing Ca(OCl)2/AgNPs composite against existing pathogenic bacteria and indicator coliform bacteria. METHODS: A total of 100 water samples were collected from the main source and water troughs used for cattle drinking and were examined for the presence of pathogenic bacteria. The susceptibility of 60 isolated strains to Ca(OCl)2, AgNPs and Ca(OCl)2/AgNPs was evaluated by the broth macrodilution method. The field trial examined different water samples collected from water supplies (pre- and post-treatment) using filter paper impregnated with Ca(OCl)2/AgNPs. RESULTS: Ca(OCl)2 loaded on AgNPs at a concentration of 1.5mg/L showed a lethal effect (100%) on Escherichia coli, Staphylococcus aureus and Klebsiella pneumoniae following 180min of exposure. Furthermore, filter paper impregnated with Ca(OCl)2/AgNPs exhibited a biocidal effect (100%) against existing pathogenic bacteria, bacterial total viable count, total coliform count and faecal coliform count. CONCLUSIONS: Use of the disinfectant Ca(OCl)2 against isolated bacteria revealed the existence of bacterial resistance. Enhancement of the biocidal effect of Ca(OCl)2 using AgNPs was reliable, proving that Ca(OCl)2/AgNPs composite had a biocidal effect against isolated bacteria at 1.5mg/L as well as inactivation of coliforms and pathogenic bacteria during percolation through bactericidal filter paper.

Assessment of carbon nanotubes and silver nanoparticles loaded clays as adsorbents for removal of bacterial contaminants from water sources.

This work evaluated the antimicrobial efficacy of kaolin clay and its loaded forms with carbon nanotubes (CNTs) and silver nanoparticles (AgNPs) against bacterial isolates from different water supplies (tap, underground and surface water) in addition to wastewater. A total of 160 water samples were collected from different water sources in the investigated districts. Samples were cultured for isolation and serological identification of pathogenic bacteria. AgNPs were synthesized by a typical one-step synthesis protocol, where CNTs were carried out in a reactor employing the double bias-assisted hot filament chemical vapor deposition method. Both were characterized using transmission electron microscopy, infrared and X-ray fluorescence (XRF) spectroscopy. The antimicrobial efficacy of each of natural kaolin clay, AgNPs- and CNTs-loaded clays were evaluated by their application in four concentrations (0.01, 0.03, 0.05 and 0.1 ppm) at different contact times (5 min, 15 min, 30 min and 2 h). AgNPs-loaded clays at concentrations of 0.05 and 0.1 mg/l for 2 h contact time exhibited a higher bactericidal efficacy on Escherichia coli and Salmonella spp. (70, 70, 80 and 90%, respectively) compared to CNTs-loaded clay. Concluding, the application of AgNPs-loaded clay for removal of water bacterial contaminants at a concentration of 0.1 ppm for 2 h contact times resulted in highly effective removals.

Ecological study on antimicrobial-resistant zoonotic bacteria transmitted by flies in cattle farms.

Flies were qualitatively and quantitatively monitored on both livestock animals and the surrounding environment to investigate their role as a potential carrier for antimicrobial-resistant bacteria of zoonotic importance in cattle farms. This was done by the use of visual observations and animal photography; meanwhile, in the surrounding environment, flies were collected using sticky cards and then microscopically identified. Representative fly samples were cultured for bacterial isolation, biochemical identification, and then tested against common 12 antibiotics. The total average of dipterous flies in examined farms was 400.42 ± 6.2. Culicoides biting midges were the most common existing species (70.01 %) followed by house flies, stable flies, and mosquitoes (18.31, 7.74, and 3.91 %, respectively) at X (2) = 9.0, P < 0.05. The most predominant bacterial isolates were Escherichia coli (22.6 %), Staphylococcus aureus and Enterobacter (17.3 % each), coagulase-negative Staphylococci (CNS) (14.7 %), Klebsiella sp. (8 %), Salmonella spp. (6.7 %), and Shigella spp. and Proteus spp. (6.7 % each). The tested bacterial isolates were resistant to variant antibiotics used. S. aureus exhibited 100 % resistance to colistine. However, E. coli revealed 92.9 and 78.6 % resistance against tetracycline and colistine, respectively. Both Salmonella spp. and Shigella spp. were 100 % resistant to penicillin, and Klebsiella sp. had 100 % resistance to tetracycline. In conclusion, Culicoides biting midges and house flies could be considered as a potential carrier for multi-drug-resistant bacteria of zoonotic importance. Furthermore, cows' environment has an essential role in propagation and wide spread of antimicrobial-resistant bacterial pathogens.

Impact of Seasonal Conditions on Quality and Pathogens Content of Milk in Friesian Cows.

Heat stress negatively affects milk quality altering its nutritive value and cheese making properties. This study aimed at assessing the impact of seasonal microclimatic conditions on milk quality of Friesian cows. The study was carried out in a dairy farm from June 2013 to May 2014 at Beni-Suef province, Egypt. Inside the barn daily ambient temperature and relative humidity were recorded and used to calculate the daily maximum temperature-humidity index (mxTHI), which was used as indicator of the degree of heat stress. The study was carried out in three periods according to the temperature-humidity index (THI) recorded: from June 2013 to September 2013 (mxTHI>78), from October 2013 to November 2013 (mxTHI 72-78) and from December 2013 to April 2014 (mxTHI<72). Eighty Friesian lactating dairy cows were monitored in each period. The three groups of cows were balanced for days in milk and parity. Milk quality data referred to somatic cell count, total coliform count (TCC), faecal coliform count (FCC), Escherichia coli count, percentage of E. coli, and Staphylococcus aureus, percentage of fat, protein, lactose, total solid and solid non-fat. Increasing THI was associated with a significant decrease in all milk main components. An increase of TCC, FCC, and E. coli count from mxTHI<72 to mxTHI>78 was observed. In addition, the isolation rate of both S. aureus and E. coli increased when the mxTHI increased. The results of this study show the seriousness of the negative effects of hot conditions on milk composition and mammary gland pathogens. These facts warrant the importance of adopting mitigation strategies to alleviate negative consequences of heat stress in dairy cows and for limiting related economic losses.

Field study on evaluation of the efficacy and usability of two disinfectants for drinking water treatment at small cattle breeders and dairy cattle farms.

The hygienic quality of drinking water for cattle originated from different sources together with the efficacy and usability of two types of disinfectants against waterborne pathogens were assessed for small cattle breeders and dairy cattle farms. A total of 120 drinking water samples were collected from water troughs representing three different water sources commonly used for cattle drinking (tap, underground and surface water; n = 65, 25, and 30, respectively). Collected samples were cultured for isolation and identification of pathogenic bacteria using serological techniques and PCR. The bactericidal efficacy of the disinfectants, sodium dichloroisocyanurate (NaDCC) and hydrogen peroxide (H2O2) 50%, at different concentrations were evaluated by the determination of total viable and coliform counts of water prior and postwater treatment. In small cattle breeders, Escherichia coli was the most prevalent bacterial isolates from surface water (56.7%) followed by Staphylococcus aureus (36.7%), Salmonella spp. (26.7%), Streptococcus faecalis (23.3%), Shigella flexneri (16.7%), Proteus spp. (16.7%), and Klebsiella pneumonae (10.0 %) at X(2) = 9, P ≤ 0.01. Prior to the use of disinfectants, the averages of total bacterial and coliform counts were the highest in surface water (3.56 × 10(7), 240.0, and 38.0 CFU/100 ml, respectively). It has been found that hydrogen peroxide 50% at a concentration of 35 mg/l had a lethal effect (100 %) on indicator microorganisms compared with NaDCC at concentration of 2 mg/l. In conclusion, the higher bacterial contaminants in drinking water were found in surface water followed by tap water, particularly for small cattle breeders. Therefore, the usage of more hygienic water troughs with their regular treatment by hydrogen peroxide 50% at concentration of 35 mg/l is highly recommended to control waterborne bacteria and consequently improve and maintain the animal health.