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The incrementing scope of pathogenic resistance to antibiotics has encouraged the search for antivirulence natural extracts. Therefore, our study designed to demonstrate the antimicrobial activity of an aqueous-garlic and thyme oil extracts against Gram-positive (Staphylococcus aureus) and Gram-negative (Salmonella spp.) bacteria by evaluating the influence of sub-inhibitory concentrations on the expression of the most critical virulence genes of the tested isolates.The antibacterial potential of both herbs was checked by the agar well diffusion method and minimum inhibitory concentration (MIC) assay. Interestingly, all isolates were inhibited by both extracts up to 50% concentration. Also, the MIC values of garlic extract (0.125-1µg/ml) against Salmonella isolates were lower than the values of thyme extract (0.5-8µg/ml). But in S. aureus isolates, the MIC values of thyme extract (0.25-2µg/ml) were the lowermost. Conventional PCR investigated that all S. aureus isolates carried the hlg (hemolysin) and icaA (intracellular adhesion) genes, but only six Salmonella isolates (three S. typhimurium and one each of S. kentucky, S. anatum, and S. lagos) had both the sopB (Salmonella outer protein B) and mgtC (membrane protein) genes. Real-time RT-PCR assays were performed to evaluate the extract's effect on the virulence genes. The thyme-oil extract has significantly repressed S. aureus virulence genes expression more than aqueous-garlic extract, which later one has effectively more than thyme-oil extract in downregulating the Salmonella virulence genes. In conclusion, garlic and thyme extracts can be used not only as a flavor, but also as potential antimicrobial agents against Gram-positive and negative bacteria.

Antimicrobial resistance poses a great threat and challenge to humanity. Therefore, the search for alternative ways to target and eliminate microbes from plant, animal, and marine microorganisms is one of the world’s concerns today.

| Listeriosis is classified as the third main cause of food-borne diseases and it is one of the significant bacterial zoonotic infections causing high fatality rates worldwide. The aim of this research was to isolate and characterize Listeria monocytogenes (L. monocytogenes) from different food items in Sharkia Province, Egypt. A total of 200 food samples were collected aseptically from local markets and subjected to the phenotypic and genotypic characterization of L. monocytogenes. Listeria species were isolated from 28 (56%), 9 (18%), 3 (6%) and 5 (10%) of the minced meat, poultry meat, tilapia fish, and raw milk samples, respectively. Overall, L. monocytogenes were isolated from 12 and 8 samples from urban and rural areas with percentages of 6 and 4%, respectively. Of interest, inlA, actA, prfA, and hlyA virulence genes were detected in all 20 L. monocytogenes isolates by multiplex PCR. In conclusion, L. monocytogenes is an organism of public health implications, and its recovery from the food samples sold at retail outlets indicates a breach of quality assurance.

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Recently, cellulose nanocrystals (CNs) have attracted wide attention owing to their superior properties compared to their bulk materials. For example, they represent an outstanding model for fabricating green metallic/metal oxide nanoparticles (NPs). In this study, two CNs (carboxylated CNs and sulfated CNs) extracted from agro-wastes of palm sheath fibers were used as templates for the facile and green synthesis of ZnO NPs by employing the sono-co-precipitation method. The obtained nanomaterials were characterized using TEM, EDX, UV–visible, DLS, FT-IR, and XRD analysis. As a result, the size and concentration of synthesized ZnO NPs were inversely proportional to one another and were affected by the CNs utilized and the reaction temperature used. Contagious diseases incited by multifarious toxigenic bacteria present severe threats to human health. The fabricated bio-nanocomposites were evaluated in terms of their antimicrobial efficacy by agar well diffusion method and broth microdilution assay, showing that CN–ZnO bio-nanocomposites were effective against the tested Gram-negative (Escherichia coli and Salmonella) and Gram-positive (Listeria monocytogenes and Staphylococcus aureus) bacteria. The influence of the subinhibitory concentrations of these suspensions on the expression of the most critical virulence toxin genes of the tested strains was effective. Significant downregulation levels were observed through toxigenic operons to both fabricated CN–ZnO bio-nanocomposites with a fold change ranging from 0.004 to 0.510, revealing a decline in the capacity and virulence of microorganisms to pose infections. Therefore, these newly fabricated CNS–ZnO bio-nanocomposites could be employed rationally in food systems as a novel preservative to inhibit microbial growth and repress the synthesis of exotoxins.

The study investigated 110 Enterobacteriaceae isolates from broiler chickens isolated from Sharkia poultry farms and analyzed the isolates antimicrobial resistance and the presence of integrons as a potential basis for this resistance. Antibiotic susceptibilities against 12 different antibiotics were determined by the disk diffusion method. Prevalences and classes of integrons were then detected in multi-drug resistant (MDR) strains using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) followed by sequencing of the variable parts. Fifty-three isolates were MDR (resistant to three or more antimicrobial agents). High resistance was detected for rifamycin (82.7%), erythromycin (67.2%), and amoxicillin-clavulanic acid (63%). Classes 1 and 2 integrons were detected in 38 of 53 MDR Enterobacteriaceae isolates of which the most common were Salmonella species (n=19), followed by Escherichia coli (12), Klebsiella pneumoniae (3), Proteus species (3), and Citrobacter freundii (1). Three isolates only harbored class 1 integrons while the remaining 35 isolates carried class 2.  All class 1 integron positive isolates exhibited the same gene cassettes arrangements: 1.) dfrA12-orfF–aadA27 (1.6 kbp); 2.) aadA23 (1.0 kbp); and 3.) dfrA15 (0.8 kbp). Moreover, four different gene cassettes were identified within class 2 integrons: 1.) dfrA1-sat2-aadA30 (2 kbp) in all isolates; 2.) sat2-aadA1 (1.7 kbp) in only one isolate; 3.) catB2 (0.9 kbp) in four isolates; and 4.) a new variant of sat2 (0.65 kbp) in three isolates. Efforts should be made to introduce surveillance programs for monitoring antimicrobial resistance that could potentially be transmitted from broiler chickens to human via integrons.

Caseous lymphadenitis (CLA) is a bacterial infection caused by Corynebacterium pseudotuberculosis (C. pseudotuberculosis) that affects sheep and goats, leading to abscess formation in their lymph nodes. The present study aimed to isolate and identify C. pseudotuberculosis from CLA in smallholder sheep and goats, and determine the resistance patterns, virulence, and resistance genes of the isolates. Additionally, genotypic and phylogenetic analysis of the isolates was conducted using ERIC-PCR and DNA sequencing techniques. A cross-sectional study examined 220 animals (130 sheep and 90 goats) from 39 smallholder flocks for clinical signs of CLA. Fifty-four (24.54%) animals showed CLA-compatible lesions, confirmed by C. pseudotuberculosis isolation and PCR identification. Sheep had a lower infection rate of CLA (18.46%) compared with goats (33.3%). Antimicrobial susceptibility testing of 54 C. pseudotuberculosis isolates to 24 antimicrobial drugs revealed that they were 100% resistant to bacitracin and florfenicol, while none of the isolates were resistant to norfloxacin. A high resistance rate was observed for penicillin and erythromycin (92.6% each). Interestingly, 16.7% of C. pseudotuberculosis isolates recovered from sheep showed vancomycin resistance. Molecular characterization of C. pseudotuberculosis isolates revealed that PLD, PIP, and FagA virulence genes were present in all examined isolates. However, the FagB, FagC, and FagD genes were detected in 24 (100%), 20 (83%), and 18 (75%) of the sheep isolates, and 26 (87%), 26 (87%), and 18 (60%) of the goat isolates, respectively. The β-lactam resistance gene was present in all isolates. Furthermore, 83% of the sheep isolates carried the aminoglycoside (aph(3″)-lb), chloramphenicol (cat1), and bacitracin (bcrA) resistance genes. Among the isolates recovered from goats, 73% were found to contain macrolides (ermX), sulfonamide (sul1), and bacitracin (bcrA) resistance genes. It is worrisome that the glycopeptide (vanA) resistance gene was detected in 8% of the sheep isolates as a first report. ERIC-PCR genotyping of 10 multi-drug-resistant C. pseudotuberculosis isolates showed a high similarity index of 83.6% between isolates from sheep and goats. Nucleotide sequence analysis of partial 16S rRNA sequences of C. pseudotuberculosis revealed 98.83% similarity with biovar Ovis of globally available reference sequences on the Genbank database. Overall, our findings might indicate that C. pseudotuberculosis infection in smallholders in Egypt might be underestimated despite the significant financial impact on animal husbandry and potential health hazards it poses. Moreover, this study highlights the importance of implementing a sustainable control strategy and increasing knowledge and awareness among smallholder breeders to mitigate the economic impact of CLA.