Extended-spectrum ß-lactamase-producing E. coli from retail meat and workers: genetic diversity, virulotyping, pathotyping and the antimicrobial effect of silver nanoparticles.

BACKGROUND: The spread of extended-spectrum ß-lactamases (ESBL) producing E. coli from food animals and the environment to humans has become a significant public health concern. The objectives of this study were to determine the occurrence, pathotypes, virulotypes, genotypes, and antimicrobial resistance patterns of ESBL-producing E. coli in retail meat samples and workers in retail meat shops in Egypt and to evaluate the bactericidal efficacy of silver nanoparticles (AgNPs-H2O2) against multidrug resistant (MDR) ESBL-producing E. coli. RESULTS: A total of 250 retail meat samples and 100 human worker samples (hand swabs and stool) were examined for the presence of ESBL- producing E. coli. Duck meat and workers' hand swabs were the highest proportion of ESBL- producing E. coli isolates (81.1%), followed by camel meat (61.5%). Pathotyping revealed that the isolates belonged to groups A and B1. Virulotyping showed that the most prevalent virulence gene was Shiga toxin 2 (stx2) associated gene (36.9%), while none of the isolates harbored stx1 gene. Genotyping of the identified isolates from human and meat sources by REP-PCR showed 100% similarity within the same cluster between human and meat isolates. All isolates were classified as MDR with an average multiple antibiotic resistance (MAR) index of 0.7. AgNPs-H2O2 at concentrations of 0.625, 1.25, 2.5 and 5 µg/mL showed complete bacterial growth inhibition. CONCLUSIONS: Virulent MDR ESBL-producing E. coli were identified in retail meat products in Egypt, posing significant public health threats. Regular monitoring of ESBL-producing E. coli frequency and antimicrobial resistance profile in retail meat products is crucial to enhance their safety. AgNPs-H2O2 is a promising alternative for treating MDR ESBL-producing E. coli infections and reducing antimicrobial resistance risks.

Irradiation as a Promising Technology to Improve Bacteriological and Physicochemical Quality of Fish.

Fish is an excellent source of protein and other essential minerals and vitamins; nevertheless, several food-borne disease outbreaks have been linked to the consumption of different types of fish. Therefore, we aimed to overcome these health threats by evaluating gamma radiation as a good fish preservation method. The aerobic plate count (APC), identification of most common pathogenic bacteria, organoleptic properties, proximate composition, and other chemical evaluations were detected in both untreated and gamma-treated fish. The overall grades of organoleptic evaluations ranged from good to very good. Fortunately, the overall chemical analysis of all examined fish samples was accepted. For the untreated fish samples, the APC was within and above the permissible limit (5 × 107 CFU/g). Pathogenic bacteria were detected with a high prevalence rate, especially S. aureus, which was found in high percentages among examined untreated fish samples. Regarding the treated fish samples, APC and pathogenic bacterial counts were reduced in a dose-dependent manner, and the irradiation at dose 5 KGy resulted in complete eradication of the aerobic plate count (not detectable) with a mean reduction percentage equal to 100%. However, gamma irradiation has no significant effect on proximate composition; particularly, carbohydrates, proteins, and lipids were not significantly affected by low and medium doses of radiation. Therefore, gamma irradiation is a highly effective fish preservation method without any effect on fish quality. Additionally, gamma irradiation as a cold process is an attractive technology for solving the problem arising from fish-borne pathogens, and it has been purposed in this study as a cheap and safe method for reducing microbial contamination of fish.

Physical and biological changes of copper oxide and hydroxyapatite filled in polycaprolactone scaffolds: Cellular growth behavior and antibacterial activity.

Burns have placed a devastating burden on public health because of leading to an increased risk of infection. Therefore, the development of an effective antibacterial dressing for wound healing is essential. The present work is mainly based on the fabrication of biodegradable polycaprolactone (PCL) films through a simple and cheap process of polymer casting using a novel combination of hydroxyapatite (HAP), cuprous oxide (Cu2O) NPs and graphene oxide (GO) nanosheets which have a great effect in preventing colonization and to modify the wound dreasing. The compositions played a key role in decreasing the contact angle of PCL from 47.02° to 11.53°. Further, the cell viability exhibited a viable cell ratio of 81.2% after 3 days of culturing. Moreover, the highest antibacterial activity was obtained from the film of Cu2O@PCl and showed high impact results in antibacterial behavior.

Growth, carcass parameters, biochemical and oxidative stress indices, and meat traits of duck breeds under different stocking densities.

This investigation was aimed to inspect if there is an influence of various stocking density on growth, carcass parameters, blood indices, and meat traits of Muscovy and Mallard ducks. One hundred twenty-six 1-day-old of each Muscovy and Mallard ducks were randomly allocated into three experimental groups with different stocking density. Group one (SD1) was 5 ducks/m2, while group 2 (SD2) was 7 ducks/m2 and group 3 (SD3) was 9 ducks/m2. The growth, carcass parameters, meat quality, blood indices were calculated. Body weight of SD1 was 18 and 4.5% heavier than SD2, while, it was 29.5 and 12% heavier than SD3 of Muscovy and Mallard duck breeds, respectively. SD3 possessed the highest levels of, H/L, ALT, AST, LDL, VLDL, and MDA with the lowest levels of lymphocyte, SOD,GSH, GPX, C3, total antioxidant capacity and IGG of both ducks' breeds. The carcass weight decreased by 40 and 15% from SD1 to SD3 in Muscovy and mallard ducks, respectively. The dressing % was highest at SD1 (84 and 83%) when compared with SD3(71and80%) of Muscovy and Mallard ducks, respectively. Cooking loss was 20 and 16% greater in group three when compared with group one in Muscovy and Mallard ducks, respectively. In conclusion ducks raised in low SD possessed the best performance with better welfare.

Evaluation of the Antimicrobial Activity of Four Plant Essential Oils against Some Food and Phytopathogens Isolated from Processed Meat Products in Egypt.

Synthetic preservatives are widely utilized by the food industry to inhibit the microbial contamination and increase food safety and shelf life. The excessive utilization of synthetic preservatives can have a negative impact on human health and the environment. There is a great interest to find out natural substances as possible food-preservatives. The consumers' preference for food products with natural ingredients prompted food manufacturers to utilize natural-based preservatives in their production. It is worth noting that plant essential oils (EOs) among the natural-based substances have been efficiently used as antimicrobial agents against phyto- and food pathogens. The current study was conducted to evaluate the microbial contamination of three industrial meat products from five governorates in Egypt, identify the predominant bacterial and fungal isolates and determine the antimicrobial efficacy of some EOs (thyme, fennel, anise and marjoram) against the most predominant microbial isolates. A sensory test was also performed to estimate the customer preferences for specific organoleptic aspects of meat products after EOs treatment. Results showed that there is a promising antimicrobial activity of all studied EOs against some microbial isolates in a dose-dependent manner. In particular, thyme EO showed the highest significant antibacterial activity against P.fluorescence and E. coli. Whereas the marjoram EO showed the highest activity against P. aeruginosa. In addition, the sensory test revealed that the treatment with anise and marjoram EOs showed the highest acceptability by the testers and did not show significant differences on the organoleptic properties with respect to control. As overall, the obtained results of the current research are promising and proved feasibility of employing plant EOs as possible preservatives for processed meat products.

Anti-Biofilms' Activity of Garlic and Thyme Essential Oils against Salmonella typhimurium.

Biofilm control by essential oil (EO) application has recently increased to preclude biofilm production on foods and environmental surfaces. In this work, the anti-biofilm effects of garlic and thyme essential oils using the minimum inhibitory concentration (MIC) method against Salmonella typhimurium recovered from different abattoir samples were investigated along with the virulence genes (InvA, SdiA and Stn genes), and the antimicrobial susceptibility profile of S. typhimurium as well. The obtained results revealed that S. typhimurium contaminated abattoir samples to varying degrees. The InvA gene was investigated in all isolates, whereas the SdiA and Stn genes were observed in four and three isolates, respectively. Utilizing the disc diffusion method, S. typhimurium isolates demonstrated substantial resistance to most of the examined antibiotics with a high multiple antibiotic resistance index. S. typhimurium isolates demonstrated biofilm formation abilities to various degrees at varied temperatures levels (4 °C and 37 °C). In conclusion, the obtained samples from the research area are regarded as a potential S. typhimurium contamination source. Furthermore, garlic essential oil (GEO) has more potential to inhibit S. typhimurium biofilm at different sub-minimum inhibitory concentrations as compared to thyme essential oil (TEO). Therefore, these EOs are considered as potential natural antibacterial options that could be applied in food industry.

Genetic Relatedness, Antibiotic Resistance, and Effect of Silver Nanoparticle on Biofilm Formation by Clostridium perfringens Isolated from Chickens, Pigeons, Camels, and Human Consumers.

In this study, we determined the prevalence and toxin types of antibiotic-resistant Clostridium perfringens in chicken, pigeons, camels, and humans. We investigated the inhibitory effects of AgNPs on biofilm formation ability of the isolates and the genetic relatedness of the isolates from various sources determined using RAPD-PCR. Fifty isolates were identified using PCR, and all the isolates were of type A. The cpe and cpb2 genes were detected in 12% and 56% of the isolates, respectively. The effect of AgNPs on biofilm production of six representative isolates indicated that at the highest concentration of AgNPs (100 µg/mL), the inhibition percentages were 80.8-82.8%. The RAPD-PCR patterns of the 50 C. perfringens isolates from various sources revealed 33 profiles and four clusters, and the discriminatory power of RAPD-PCR was high. Multidrug-resistant C. perfringens isolates are predominant in the study area. The inhibition of biofilm formation by C. perfringens isolates was dose-dependent, and RAPD-PCR is a promising method for studying the genetic relatedness between the isolates from various sources. This is the first report of AgNPs' anti-biofilm activity against C. perfringens from chickens, pigeons, camels, and humans, to the best of our knowledge.

Molecular characterization, antibiotic resistance pattern and biofilm formation of Vibrio parahaemolyticus and V. cholerae isolated from crustaceans and humans.

Human infection with pathogenic vibrios is associated with contaminated seafood consumption. In the present study, we examined 225 crustaceans collected from retail markets in Egypt. Stool samples from gastroenteritis patients were also examined. Bacteriological and molecular examinations revealed 34 (15.1%) V. parahaemolyticus and 2 (0.9%) V. cholerae from crustaceans, while V. parahaemolyticus isolates were identified in 3 (3%) of the human samples. The virulence-associated genes tdh and/or trh were detected in 5.9% and 100% of the crustacean and human samples, respectively, whereas the two V. cholerae isolates were positive for the ctx and hlyA genes. Antibiotic sensitivity revealed high resistance of the isolates to the used antibiotics and an average MAR index of 0.77. Biofilm formation at different temperatures indicated significantly higher biofilm formation at 37 °C and 25 °C compared with 4 °C. Frequent monitoring of seafood for Vibrio species and their antibiotic, molecular and biofilm characteristics is essential to improve seafood safety.

Occurrence, Virulence Factors, Antimicrobial Resistance, and Genotyping of Staphylococcus aureus Strains Isolated from Chicken Products and Humans.

Staphylococcus aureus in food is a consequence of inadequate hygienic handling and processing, posing a potential risk to public health. The current study aimed to characterize virulence factors, as well as antimicrobial resistance of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) isolated from retail chicken products and hand swabs from vendors in Egypt. In addition, genetic relatedness of the isolates from chicken and humans was evaluated by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using protein A as a target. A total of 110 samples were collected from chicken products (n = 80) and vendors (n = 30). Overall, 30 (37.5%) chicken products samples were positive for S. aureus, whereas hand swabs from meat handlers revealed that 18 (60%) were positive. Ten MRSA strains were characterized by the presence of the mecA gene, comprising seven isolates from chicken and three from humans. Virulence-associated factors were evaluated by PCR, revealing that 31.3% of S. aureus isolates harbored the Panton-Valentine leukocidin (PVL) gene, whereas 10.4% were positive for the sea and sed genes each, and only two isolates were positive for γ-hemolysin-associated gene. Genotyping using spa PCR-RFLP showed identical restriction banding patterns of MRSA isolates of human and chicken meat origin, indicating the genetic relatedness of the isolates. To the best of our knowledge, this is the first study to characterize PVL-positive MRSA from chicken products and to utilize spa-RFLP for evaluating the genetic relatedness between MRSA of human and chicken origin in Egypt.