Whole-genome sequencing of Listeria innocua recovered from retail milk and dairy products in Egypt.

The similarity of the Listeria innocua genome with Listeria monocytogenes and their presence in the same niche may facilitate gene transfer between them. A better understanding of the mechanisms responsible for bacterial virulence requires an in-depth knowledge of the genetic characteristics of these bacteria. In this context, draft whole genome sequences were completed on five L. innocua isolated from milk and dairy products in Egypt. The assembled sequences were screened for antimicrobial resistance and virulence genes, plasmid replicons and multilocus sequence types (MLST); phylogenetic analysis of the sequenced isolates was also performed. The sequencing results revealed the presence of only one antimicrobial resistance gene, fosX, in the L. innocua isolates. However, the five isolates carried 13 virulence genes involved in adhesion, invasion, surface protein anchoring, peptidoglycan degradation, intracellular survival, and heat stress; all five lacked the Listeria Pathogenicity Island 1 (LIPI-1) genes. MLST assigned these five isolates into the same sequence type (ST), ST-1085; however, single nucleotide polymorphism (SNP)-based phylogenetic analysis revealed 422-1,091 SNP differences between our isolates and global lineages of L. innocua. The five isolates possessed an ATP-dependent protease (clpL) gene, which mediates heat resistance, on a rep25 type plasmids. Blast analysis of clpL-carrying plasmid contigs showed approximately 99% sequence similarity to the corresponding parts of plasmids of L. monocytogenes strains 2015TE24968 and N1-011A previously isolated from Italy and the United States, respectively. Although this plasmid has been linked to L. monocytogenes that was responsible for a serious outbreak, this is the first report of L. innocua containing clpL-carrying plasmids. Various genetic mechanisms of virulence transfer among Listeria species and other genera could raise the possibility of the evolution of virulent strains of L. innocua. Such strains could challenge processing and preservation protocols and pose health risks from dairy products. Ongoing genomic research is necessary to identify these alarming genetic changes and develop preventive and control measures.

Prevalence, antibiotic resistance patterns, and biofilm formation ability of Enterobacterales recovered from food of animal origin in Egypt.

Background and Aim: The majority of animal-derived food safety studies have focused on foodborne zoonotic agents; however, members of the opportunistic Enterobacteriaceae (Ops) family are increasingly implicated in foodborne and public health crises due to their robust evolution of acquiring antimicrobial resistance and biofilms, consequently require thorough characterization, particularly in the Egyptian food sector. Therefore, this study aimed to determine the distribution and prevalence of Enterobacteriaceae family members in animal-derived foods, as well as their resistance to important antimicrobials and biofilm-forming potential. Materials and Methods: A total of 274 beef, rabbit meat, chicken meat, egg, butter, and milk samples were investigated for the presence of Enterobacteriaceae. All isolated strains were first recognized using traditional microbiological techniques. Following that, matrix-assisted laser desorption ionization-time of flight mass spectrometry was used to validate the Enterobacteriaceae's identity. The isolated enterobacteria strains were tested on disk diffusion and crystal violet quantitative microtiter plates to determine their antibiotic resistance and capacity to form biofilms. Results: There have been thirty isolates of Enterobacteriaceae from seven different species and four genera. Out of the three food types, Pseudomonas aeruginosa had the highest prevalence rate (4.1%). With three species, Enterobacter genera had the second-highest prevalence (3.28%) across five different food categories. In four different food types, the Klebsiella genera had the second-highest distribution and third-highest incidence (2.55%). Almost all isolates, except three Proteus mirabilis, showed prominent levels of resistance, particularly to beta-lactam antibiotics. Except for two Enterobacter cloacae and three P. mirabilis isolates, all isolates were classified as multidrug-resistant (MDR) orextensively multidrug-resistant (XDR). The multiple antibiotic resistance index (MARI) of the majority of isolates dropped between 0.273 and 0.727. The highest MARI was conferred by Klebsiella pneumoniae, at 0.727. Overall, 83.33% of the isolates had strong biofilm capacity, while only 16.67% exhibited moderate capacity. Conclusion: The MDR, XDR, and strong biofilm indicators confirmed in 83.33% of the currently tested Enterobacteriaceae from animal-derived foods suggest that, if not addressed, there may be rising risks to Egypt's economy and public health.

Prevalence and molecular characterization of foodborne and human-derived Salmonella strains for resistance to critically important antibiotics.

The primary goals of this cross-sectional study were to screen various food/water, and human samples for the presence of Salmonella species, and to assess the phenotypic and genetic relationship between resistances found in food and human Salmonella isolates to critically important antibiotics. Between November 2019 and May 2021, 501 samples were randomly collected for Salmonella isolation and identification using standard culturing methods, biochemical, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and PCR techniques. Antimicrobial susceptibility testing was performed on confirmed Salmonella species, and PCR was used to investigate the genetic components that confer these resistance traits. Salmonella enterica subspecies enterica was confirmed in 35 (6.99%) of the samples (raw food = 23, ready-to-eat food/drink [REF/D] = 5, human = 7). Seventeen of them were antibiotic-resistant to at least one class, and eight were multidrug-resistant (MDR) isolates (raw food = 7, human = 1). All Salmonella isolates were susceptible to carbapenems, third- and fourth-generation cephalosporins and monobactam antibiotics. Resistance phenotypes to aminoglycosides (48.57%), ß-lactams (20%) and tetracycline (17.14%), as well as associated genes such as aadA, blaTEM , blaZ and tetA, as well as dfrA and sul1, were prevalent in Salmonella isolates. Colistin resistance genotype (mcr1) was detected in three (8.57%) isolates recovered from egg, cattle mince and rabbit meat, and the total incidence was 14.29% when two isolates exhibited resistance phenotypes were considered. Furthermore, four (11.43%) MDR isolates shared the blaTEM and blaZ genes, and one (2.86%) isolate contained three extended spectrum ß-lactams producing genes (ESBL), namely blaCTX , blaTEM and blaZ . The gyrA gene was expressed by one of three foodborne Salmonella isolates (8.57%) with ciprofloxacin resistance phenotypes. To the best of our knowledge, this is the first report from Egypt identifying colistin resistance in Salmonella enterica recovered from cattle minced meat and rabbit meat. Overall, the highest incidence rate of Salmonella enterica was found in cattle-derived products, and it was slightly more prevalent in RTE/D foods than in raw foods. Resistance to critical and clinically important antibiotics, particularly in Salmonella from RTE/D food, suggests that these antibiotics are being abused in the investigated area's veterinary field, and raises the potential of these isolates being transmitted to high-risk humans, which would be a serious problem. Future research using whole-genome sequencing is needed to clarify Salmonella resistance mechanisms to critically important antimicrobial agents or those exhibiting multidrug resistance.

Impact of Fennel Essential Oil as an Antibiotic Alternative in Rabbit Diet on Antioxidant Enzymes Levels, Growth Performance, and Meat Quality.

In the current study, fennel essential oil was used as an antibiotic alternative compared to gentamycin for enhancing the expression of apoptosis genes and antioxidant enzymes in weaned rabbits as well as meat quality and growth performance. The gene expression of the cell lymphoma 2 (BAX and BCL2), caspase3 (CASP3), and glutathione peroxidase (GPX1) were estimated in the liver tissue using qRT-PCR. A total of 45 Moshtohor weaned male rabbits aged four weeks were randomly allocated to control, T1, and T2 treatment groups; each consisted of 15 weaned male rabbits with five replicates. Rabbits in the T1 and T2 groups were orally supplied with 1 mL fennel oil and 1 mL gentamycin, respectively. Weaned rabbits under different treatments showed increased body weight (BW) at 8 and 12 weeks of age and average daily gain (ADG) at 4-8 and 4-12 weeks of age compared to the control group. Compared to the controls, the weaned rabbits supplemented with fennel oil and gentamycin had lower total cholesterol, triglyceride, and MDA. In addition, villus length, mRNA of BAX, BCL2, Casp3, and GPX were increased in the different treatments compared to the control. Furthermore, the meat of these rabbits was less tender, had a lower aerobic plate count (APC), pH, and was brighter and redder in color than the control. Under the conditions of the present study, the supplementation of weaned Moshtohor rabbits with fennel oil as a natural alternative for gentamycin enhanced feed conversion and daily gain through enhancing villus length and mucus thickness. Additionally, fennel essential oil reduces oxidative stress by increasing the antioxidant enzymes.

Health Aspects, Growth Performance, and Meat Quality of Rabbits Receiving Diets Supplemented with Lettuce Fertilized with Whey Protein Hydrolysate Substituting Nitrate.

Lettuce (Lactuca sativa) was grown using a foliar spray with whey protein hydrolysate (WPH) as opposed to normal nitrate fertilization. Lettuce juice was prepared from lettuce cultivated without any fertilization, nitrate fertilization, or WPH. Sixty weaned, 4-week-old male V-line rabbits with an average 455 ± 6 g body weight were randomly divided into 4 groups (n = 15) and administered different lettuce juices. Rabbits administered WPH-fertilized lettuce showed significantly higher (n = 5, p < 0.05) body weight and carcass weight than those receiving nitrate-fertilized lettuce. Rabbits administered nitrate-fertilized lettuce were associated with significantly (p < 0.05) higher levels of liver enzyme activities (AST, ALT, and ALP), bilirubin (total, direct, and indirect), and kidney biomarkers (creatinine, urea, and uric acid). Rabbits administered WPH-fertilized lettuce avoided such increases and exhibited normal levels of serum proteins. Rabbits administered nitrate-fertilized lettuce manifested significantly (p < 0.05) lower RBCs and Hb levels than that of the other groups, while those receiving WPH-fertilized lettuce showed the highest levels. Liver and kidney sections of rabbits receiving WPH-fertilized lettuce witnessed the absence of the histopathological changes induced by feeding on nitrate-fertilized lettuce and produced higher quality meat. WPH-lettuce can substitute nitrate-fertilized lettuce in feeding rabbits for better performance and health aspects.

Biosynthesized Iron Oxide Nanoparticles from Petroselinum crispum Leaf Extract Mitigate Lead-Acetate-Induced Anemia in Male Albino Rats: Hematological, Biochemical and Histopathological Features.

This study aimed to investigate the ameliorative effects of iron oxide nanoparticles (IONPs) prepared from leaf extract of Petroselinum crispum compared to those prepared using a chemical method in lead-acetate-induced anemic rats. Twenty rats were divided into four groups (five rats each). Throughout the experimental period (8 weeks), the rats in group 1 were not given any therapy. The rats in groups 2, 3 and 4 were given 400 ppm lead acetate orally for 2 weeks to make them anemic. Following that, these rats were either left untreated, given 27 ppm of chemical IONPs orally or given 27 ppm of natural IONPs orally for the remaining 6 weeks of the experiment. TEM analysis indicated that the chemically and naturally prepared IONPs had sizes of 6.22-9.7 and 64-68 nm, respectively. Serum ferritin and iron concentrations were reduced, whereas the total iron-binding capacity (TIBC), ALT, AST, urea and creatinine were significantly increased in the non-treated lead-acetate-induced anemic rats compared to those of the control. In addition, congestion, hemorrhage, necrosis, vacuolation and leukocytic infiltration in the kidneys, liver and spleen were observed in non-treated lead-acetate-induced anemic rats compared to the control. The effects of lead acetate were mitigated by IONPs, particularly the natural one. In conclusion, IONPs produced from Petroselinum crispum leaf extract can be used as an efficient and safe therapy in lead-acetate-induced anemic rats.

Enrichment of Broiler Chickens' Meat with Dietary Linseed Oil and Lysine Mixtures: Influence on Nutritional Value, Carcass Characteristics and Oxidative Stress Biomarkers.

This study aimed to evaluate the effect of four combinations of dietary linseed oil and lysine mixtures on performance, fatty and amino acid profiles, oxidative stress biomarkers, cell energy and meat quality parameters of broiler chickens. One hundred and sixty broiler chicks were allocated into four groups. Birds of groups 1-4 were fed diets containing optimum lysine and 2% of linseed oil, optimum lysine and 4% of linseed oil, high lysine and 2% of linseed oil, and high lysine and 4% of linseed oil, respectively, for a period of 35 days. High linseed oil or lysine levels did not affect the performance of the tested birds, but the high level of dietary linseed oil decreased the concentrations of muscles' saturated fatty acids (SFA). The highest values of ω-3 polyunsaturated fatty (ω-3 PUFA) and arachidonic acids with lowest levels of monounsaturated fatty (MUFA) were detected in the muscles of birds fed diets containing high linseed oils and/or lysine levels. High linseed oil or lysine levels provided the best essential amino acid profile and improved antioxidant components as well as cell energy, and tenderness and redness of the meat. Conclusively, high dietary lysine and linseed oil combinations improved the nutritional value, antioxidant status and cell energy of broiler chickens' meat.