Ecological prevalence, genetic diversity, and multidrug resistance of Salmonella enteritidis recovered from broiler and layer chicken farms.

Salmonella is a significant foodborne pathogen that has a significant impact on public health, and different strains of multidrug resistance (MDR) have been identified in this genus. This study used a combination of phenotypic and genotypic approaches to identify distinct Salmonella species collected from poultry broiler and layer farms, and antibiotic sensitivity testing was performed on these species. A total of 56 Salmonella isolates were serotyped, and phenotypic antibiotic resistance was determined for each strain. The enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) method was also used to provide a genotypic description, from which a dendrogram was constructed and the most likely phylogenetic relationships were applied. Salmonella isolates were detected in 20 (17%) out of 117 samples collected from small-scale broiler flocks. Salmonella isolates were classified as MDR strains after showing tolerance to 4 antibiotics, but no resistance to cloxacillin, streptomycin, vancomycin, or netilmicin was observed. From a genotypic perspective, these strains lack dfrD, parC, and blasfo-1 resistant genes, while harboring blactx-M, blaDHA-L, qnrA, qnrB, qnrS, gyrA, ermA, ermB, ermC, ermTR, mefA, msrA, tet A, tet B, tet L, tet M resistance genes. The genotyping results obtained with ERIC-PCR allowed isolates to be classified based on the source of recovery. It was determined that Salmonella strains displayed MDR, and many genes associated with them. Additionally, the ERIC-PCR procedure aided in the generation of clusters with biological significance. Extensive research on Salmonella serotypes is warranted, along with the implementation of long-term surveillance programs to monitor MDR Salmonella serotypes in avian-derived foods.

In ovo protective effects of chicoric and rosmarinic acids against Thiacloprid-induced cytotoxicity, oxidative stress, and growth retardation on newly hatched chicks.

Thiacloprid (TH) is a neonicotinoid insecticide employed in agriculture to protect fruits and vegetables against different insects. It showed different deleterious effects on the general health of non-target organisms including birds and animals, however, its developmental toxicity has yet to be fully elucidated. Chicoric (CA) and rosmarinic (RA) acids are polyphenolic compounds with a wide range of beneficial biological activities. In this study, the possible protective effects of CA and RA were investigated in chick embryos exposed in ovo to TH (1µg/egg) with or without CA (100 µg/egg) or RA (100 µg/egg) co-exposure. TH reduced the hatchling body weight, body weight/egg weight, and relative weight of bursa of Fabricius in the one-day-old hatchlings. Examination of the 7-day-old chicks revealed a decline in feed intake, daily weight gain, feed conversion ratio (FCR), and plasma levels of T3, T4, and growth hormone. Serum ALT, AST activities, and total cholesterol levels showed significant elevations. Hepatic MDA was increased with a reduction in SOD activity and GSH level and downregulation of the liver SOD and GST gene expression pattern. Serum IgG and IgM levels were reduced, and various histopathological alterations were noticed in the liver. Co-administration of CA or RA with TH mitigated the toxic effects on hatchlings. When both CA and RA are combined, they present a synergistic protective effect. CA and RA can be used as protective agents against TH toxicity as they improve growth performance and have hepatoprotective and immunostimulant effects in newly hatched chicks.

Occurrence, virulence, carbapenem resistance, susceptibility to disinfectants and public health hazard of Pseudomonas aeruginosa isolated from animals, humans and environment in intensive farms.

AIMS: This work aimed to determine the occurrence, virulence, antibiogram, carbapenem resistance genes and susceptibility to disinfectants of Pseudomonas aeruginosa isolated from animals, environment and workers in intensive farms. METHODS AND RESULTS: A total of 610 samples from intensive beef cattle and sheep farms in Kafr El Sheikh Governorate, Egypt were screened for the presence of P. aeruginosa using bacteriological assays. The isolates were characterized by PCR and tested for susceptibility to antibiotics using disk diffusion method and disinfectants by quantitative suspension test. In all, 60 P. aeruginosa isolates were recovered in this study and all isolates harboured at least one of the virulence genes tested. Human P. aeruginosa isolates were highly resistant to cephalosporins, fluroquinolones, aminoglycosides, carbapenems and penicillins+ß-lactamase inhibitors than non-human isolates. Colistin resistance was higher in non-human than human P. aeruginosa isolates, whereas low resistance to aztreonam was observed in non-human and human isolates. Carbapenem-resistant P. aeruginosa (CRPA) strains were recovered from workers (56.5%), sheep (8.3%) and cattle (8.3%). All CRPA harboured at least one of the carbapenem resistance genes tested and most of them showed multidrug resistance (MDR) or extensive drug resistance (XDR) phenotypes. Glutaraldehyde 1% and hydrogen peroxide 3% eliminated P. aeruginosa completely in the absence and presence of organic matter within short contact time compared with other disinfectants. CONCLUSIONS: This study reported the occurrence of CRPA in animals and workers in intensive farms. Glutaraldehyde and hydrogen peroxide were the most effective disinfectants against P. aeruginosa. SIGNIFICANCE AND IMPACT OF THE STUDY: The occurrence of CRPA in intensive livestock farms is a serious challenge that threatens animal and human health and increases the risk of P. aeruginosa infection in the community. Therefore, it is vital to control the spread of CRPA by banning or restricting the use of antibiotics and applying proper cleaning and disinfection protocols in livestock farms.

Adsorptivity of mercury on magnetite nano-particles and their influences on growth, economical, hemato-biochemical, histological parameters and bioaccumulation in Nile tilapia (Oreochromis niloticus).

Among toxic pollutants, Mercury (Hg) is a toxic heavy metal that induces harmful impacts on aquatic ecosystems directly and human being's health indirectly. This study confirmed the in vitro magnetic potential of magnetite Nano-Particles (Fe3O4 NPs) against waterborne Hg exposure-induced toxicity in Nile tilapia (Oreochromis niloticus). We further evaluate the safety profile of Fe3O4 NPs on fish growth, hemato-biochemical, histological parameters, bioaccumulation in muscles, and economy. Magnetite nanoparticles were characterized, adsorption loading to Hg ions was investigated, and testing different concentrations of Fe3O4 NPs (0.2, 0.4, 0.6, 0.8, and 1.0 mg/L) was applied to determine the highest concentration of adsorption. An in vivo experiment includes 120 fish with an average weight of 26.2 ± 0.26 g were randomly divided into 4 equal groups, each group had three replicates (n = 30 fish/group; 10 fish/ replicate). All groups were fed on a reference basal diet and the experiment was conducted for 30 days. The first group (G1) was allocated as a control. The second group (G2) received 1.0 mg/L aqueous suspension of Fe3O4 NPs. The third group (G3) was exposed to an aqueous solution of Hg ions at a concentration of 0.025 mg/L. Meanwhile, the fourth group (G4) acquired an aqueous suspension composed of a mixture of Hg ions and Fe3O4 NPs as previously mentioned. Throughout the exposure period, the clinical signs, symptoms, and mortalities were recorded. The Hg ions-exposed group induced the following consequences; reduced appetite resulting in reduced growth and less economic efficiency; microcytic hypochromic anemia, leukocytosis, lymphopenia, and neutrophilia; sharp and clear depletion in the immune indicators including lysozymes activity, immunoglobulin M (IgM), and Myeloperoxidase activities (MPO); significant higher levels of ALT, AST, urea, creatinine, and Superoxide dismutase (SOD); histological alterations of gill, hepatic and muscular tissues with strong expression of apoptotic marker (caspase 3); and a higher accumulation of Hg ions in the muscles. Surprisingly, Fe3O4 NPs-supplemented groups exhibited strong adsorption capacity against the Hg ions and mostly removed the Hg ions accumulation in the muscles. Also, the hematological, biochemical, and histological parameters were recovered. Thus, in order to assess the antitoxic role of Fe3O4 NPs against Hg and their safety on O. niloticus, and fill the gap of the research, the current context was investigated to evaluate the promising role of Fe3O4 NPs to prevent Hg-exposure-induced toxicity and protection of fish health, which ascertains essentiality for sustainable development of nanotechnology in the aquatic environment.

Effects of Pyrene on Human Liver HepG2 Cells: Cytotoxicity, Oxidative Stress, and Transcriptomic Changes in Xenobiotic Metabolizing Enzymes and Inflammatory Markers with Protection Trial Using Lycopene.

Pyrene is one of the major polycyclic aromatic hydrocarbons formed during heat treatment of meat and in car exhausts; however, few studies have investigated pyrene-induced adverse effects on human cell lines. This study aimed at the investigation of pyrene-induced cytotoxicity and oxidative damage in human liver HepG2 cells at environmentally relevant concentrations. Pyrene-induced changes in mRNA expression of xenobiotic metabolizing enzymes (XMEs), xenobiotic transporters, antioxidant enzymes, and inflammatory markers were investigated using real-time PCR. As a protection trial, the ameliorative effects of lycopene, a carotenoid abundantly found in tomato, were investigated. The possible mechanisms behind such effects were examined via studying the co exposure effects of pyrene and lycopene on regulatory elements including the aryl hydrocarbon receptor (Air) and elytroid 2-related factor 2 (RF). The achieved results indicated that pyrene caused significant cytotoxicity at 50 n, with a clear production of reactive oxygen species (ROS) in a dose-dependent manner. Pyrene upregulated mRNA expression of phase I enzymes including CYP1A1, 1A2, and CYP1B1 and inflammatory markers including TNFα and Cox2. However, pyrene significantly downregulated phase II enzymes, xenobiotic transporters, and antioxidant enzymes. Interestingly, lycopene significantly reduced pyrene-induced cytotoxicity and ROS production. Moreover, lycopene upregulated detoxification and antioxidant enzymes, probably via its regulatory effects on Air- and RF-dependent pathways.

Metal contamination in quail meat: residues, sources, molecular biomarkers, and human health risk assessment.

Quail meat is an emerging source of high-quality animal protein. Quails are exposed to a wide range of xenobiotics such as heavy metals. In this study, residual concentrations of four toxic metals, of significant public health importance, including cadmium (Cd), lead (Pb), arsenic (As), and nickel (Ni), were determined in edible tissues of quails. In addition, metal loads were measured in water, feed, and litter samples collected from same quail farms as possible sources for quail exposure to heavy metals. The possible use of metallothionein (MT) and heat shock protein 70 (Hsp70) as molecular biomarkers of exposure to heavy metals was further investigated. Furthermore, the dietary intake and the potential risk assessment of the examined heavy metals among children and adults were calculated. The edible tissues of quails contained high concentrations of four heavy metals (contents (ppm/ww) ranging from 0.02 to 0.32 in Cd, 0.05 to 1.96 in Pb, 0.002 to 0.32 in As, and 1.17 to 3.94 in Ni), which corresponded to the high contents of these metals in the feeds, water, and litter. MT and Hsp70 mRNA expressions showed positive correlations with the concentrations of heavy metals in tissues indicating the possibility to use these proteins as biomarkers for quail's exposure to toxic metals. Dietary intake of quail meat and risk assessment revealed potential risks especially for children after prolonged exposure to the examined metals. Thus, legislations should be established and continuous screening of metal residues should be adopted in order to reduce the toxic metal concentrations in feeds and drinking water for quails. Reduction of exposure to heavy metals subsequently would lead to minimization of exposure of such toxicants through consumption of quail meat.

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.

Non-Typhoidal Salmonella in poultry meat and diarrhoeic patients: prevalence, antibiogram, virulotyping, molecular detection and sequencing of class I integrons in multidrug resistant strains.

BACKGROUND: The worldwide increase of food-borne infections with antibiotic resistant pathogens constitutes a major public health problem. Therefore, this study aimed to determine the prevalence, antibiogram, virulence genes profiles and integron characteristics of non-typhoidal Salmonella spp. isolated from poultry meat and diarrhoeic patients in Egypt. METHODS: A total of 150 samples comprising (100 poultry meat and 50 diarrhoeic patients' stool) were examined for the presence of Salmonella spp. using culture methods followed by biochemical and serological identification of the isolates. All Salmonella strains were tested for their susceptibility to the antibiotics using disk diffusion method and screened for the presence of virulence genes and class I integrons using PCR. RESULTS: The overall prevalence of Salmonella spp. in poultry meat samples was 10 % compared to 4 % in diarrhoeic patients. All the isolates were serologically identified into Salmonella Typhimurium (seven isolates), S. Derby, S. Kiel, S. Rubislaw (one isolate, each) and untypable strains (two isolates). Antibiotic susceptibility testing showed a higher resistance of the total isolates to erythromycin and tetracycline (100 %, each), followed by amoxicillin-clavulanic acid (91.7 %), trimethoprim-sulfamethoxazole (83.3 %), streptomycin, nalidixic acid, ampicillin-sulbactam (75 %, each), gentamycin, ampicillin (66.7 %, each), chloramphenicol (58.3 %), ciprofloxacin (25 %) and ceftriaxone (16.7 %). Virulence genes profiles revealed the presence of sopB gene in five Salmonella strains isolated from poultry meat (n = 3) and humans (n = 2). Moreover, pefA was only identified in three isolates from poultry meat. On the other hand, S. Kiel and S. Typhimurium (one isolate, each) were harboring hilA and stn genes, respectively. Class 1 integrons were detected in all Salmonella spp. with variable amplicon sizes ranged from 650-3000 bp. Sequencing of these amplicons revealed the presence of gene cassettes harboring aac(3)-Id, aadA2, aadA4, aadA7, sat, dfrA15, lnuF and estX resistance genes. Nucleotide sequence analysis showed point mutations in the aac(3)-Id of S. Derby, aadA2, estX-sat genes of S. Typhimurium. Meanwhile, frame shift mutation was observed in aadA7 genes of S. Typhimurium. CONCLUSIONS: Increasing rate of antimicrobial resistance and class 1 integrons among multidrug resistant Salmonella spp. has prompted calls for the reduction of antimicrobial use in livestock to prevent future emergence of resistance.