Aquatic assessment of the chelating ability of Silica-stabilized magnetite nanocomposite to lead nitrate toxicity with emphasis to their impact on hepatorenal, oxidative stress, genotoxicity, histopathological, and bioaccumulation parameters in Oreochromis niloticus and Clarias gariepinus.

BACKGROUND: In recent years, anthropogenic activities have released heavy metals and polluted the aquatic environment. This study investigated the ability of the silica-stabilized magnetite (Si-M) nanocomposite materials to dispose of lead nitrate (Pb(NO3)2) toxicity in Nile tilapia and African catfish. RESULTS: Preliminary toxicity tests were conducted and determined the median lethal concentration (LC50) of lead nitrate (Pb(NO3)2) to Nile tilapia and African catfish to be 5 mg/l. The sublethal concentration, equivalent to 1/20 of the 96-hour LC50 Pb(NO3)2, was selected for our experiment. Fish of each species were divided into four duplicated groups. The first group served as the control negative group, while the second group (Pb group) was exposed to 0.25 mg/l Pb(NO3)2 (1/20 of the 96-hour LC50). The third group (Si-MNPs) was exposed to silica-stabilized magnetite nanoparticles at a concentration of 1 mg/l, and the fourth group (Pb + Si-MNPs) was exposed simultaneously to Pb(NO3)2 and Si-MNPs at the same concentrations as the second and third groups. Throughout the experimental period, no mortalities or abnormal clinical observations were recorded in any of the treated groups, except for melanosis and abnormal nervous behavior observed in some fish in the Pb group. After three weeks of sublethal exposure, we analyzed hepatorenal indices, oxidative stress parameters, and genotoxicity. Values of alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), urea, and creatinine were significantly higher in the Pb-intoxicated groups compared to the control and Pb + Si-MNPs groups in both fish species. Oxidative stress parameters showed a significant decrease in reduced glutathione (GSH) concentration, along with a significant increase in malondialdehyde (MDA) and protein carbonyl content (PCC) concentrations, as well as DNA fragmentation percentage in the Pb group. However, these values were nearly restored to control levels in the Pb + Si-MNPs groups. High lead accumulation was observed in the liver and gills of the Pb group, with the least accumulation in the muscles of tilapia and catfish in the Pb + Si-MNPs group. Histopathological analysis of tissue samples from Pb-exposed groups of tilapia and catfish revealed brain vacuolation, gill fusion, hyperplasia, and marked hepatocellular and renal necrosis, contrasting with Pb + Si-MNP group, which appeared to have an apparently normal tissue structure. CONCLUSIONS: Our results demonstrate that Si-MNPs are safe and effective aqueous additives in reducing the toxic effects of Pb (NO3)2 on fish tissue through the lead-chelating ability of Si-MNPs in water before being absorbed by fish.

Biofilms and efflux pump regulatory gene (mexR) in multidrug-resistant Pseudomonas aeruginosa isolated from migratory birds in Egypt.

Background and Aim: Multidrug-resistant (MDR) Pseudomonas aeruginosa is a global threat to public health. This study aimed to determine biofilms and efflux pump regulatory gene (mexR) in MDR P. aeruginosa isolates. Materials and Methods: A total of 42 fecal samples of aquatic migratory birds collected during hunting season in Egypt were evaluated for the detection of P. aeruginosa according to standard culture-based methods. The antibiotic susceptibility of P. aeruginosa strains was evaluated using disk diffusion methods. The biofilm formation ability of the isolates was phenotypically determined using a colorimetric microtitration plate assay. Polymerase chain reaction amplification was performed to detect biofilm genes (PelA and PslA) and mexR. Results: In total, 19 isolates (45.2%) were recovered from the 42 fecal samples of migratory birds. All isolates were identified as MDR P. aeruginosa, and 78.9% of the strains produced biofilms at different degrees. Molecular detection of biofilm extracellular polymeric substances revealed that PelA was the most predominant gene in the biofilm-producing isolates, followed by PslA. mexR was detected in 63.2% of MDR P. aeruginosa isolates, and its prevalence was higher in non-biofilm-producing strains (75%) than in biofilm-producing strains (60%). Conclusion: Antibiotic resistance in P. aeruginosa isolates recovered from migratory birds through various mechanisms is a major public and animal health problem. It is important to consider the significance of migratory birds in disease transmission.

Immunological status of some edible fishes exposed to parasitic infections in relation to heavy metals pollution.

In this study, six heavy metals (Pb, Fe, Cu, Ni, Cd, and Mn) have been measured in water, and muscles from mullet (Mugil cephalus), tilapia (Oreochromis niloticus), and African catfish (Clarias gariepinus) collected from Lake Manzal, Egypt. In addition, the existence of different encysted metacercariae in fish muscle with an evaluation of cell-mediated immune response in infected muscles was also investigated. Water samples generally contained less than the permissible level of heavy metals. The metal accumulation levels in muscle were: Pb > Ni > Cd > Cu > Fe > Mn. The levels of Pb and Ni in the muscles exceed the permissible limits, while the concentration of Mn varied significantly (p < 0.05) depending on fish species. Based on the estimated weekly intake in this study, the EWI values of these heavy metals are below the established Provisional Permissible Tolerable Weekly Intake. On the other hand, Prohemistomum vivax encysted metacercaria were found in the muscle of O. niloticus and C. gariepinus with the intensity of 1-10 cyst per 1 cm of muscle. While M. cephalus was found to be infected with Heterophyes heterophyes EMC. TNF- α1 was 10 folds upregulated in O. niloticus than in control fish. IL-1ß expression in O. niloticus was upregulated by 15 folds compared with the control one. By examining C. gariepinus, the MHC II gene expression was increased by 15-fold in comparison to the control group.

Sequencing and phylogenetic analysis of the stn gene of Salmonella species isolated from different environmental sources at Lake Qarun protectorate: The role of migratory birds and public health importance.

BACKGROUND AND AIM: Salmonella causes most foodborne bacterial illnesses worldwide. It is found in various hosts, including pets, farm animals, and wild animals, as well as the environment. This study aimed to examine the epidemiological relationship between Salmonella isolates from aquatic environments and those from other avian hosts. MATERIALS AND METHODS: The study examined 12 water samples, 210 aquatic animals, and 45 migratory aquatic bird samples collected from the protected area of Lake Qarun in El-Fayoum Governorate, Egypt, during migration seasons from different waterfowl migration areas (from October 2018 to January 2019). In addition, 45 fecal samples from domestic chickens were collected from the same geographic location from poultry farms. Bacteriological examination and polymerase chain reaction assay of two virulence genes (i.e., invA and stn) were performed to isolate and identify Salmonella. RESULTS: Salmonella was isolated from 58.3% (7/12) of Lake Qarun water samples, 13.3% (6/45) of migratory waterfowl, 6.6% of (3/45) of chickens (Gallus gallus domesticus), and 4.3% (3/70) of fish and pooled brine shrimp. In migratory aquatic bird species that were sampled, Salmonella were isolated from 23.1% (3/13) of Eurasian coot (Fulica atra), 12.5%, (1/8) of green-winged teal (Anas cardolinesis), 10% (2/20) of northern shoveler (Spatula clypeata), and 0% (0/4) of mallard duck (Anas platyrhynchos). In 35 Tilapia, Salmonella was isolated by (8.6%) 5.7% of external surfaces, 2.85% from the intestine, and 0% from the muscle. No Salmonella was isolated from the 175 brine shrimp samples. Phylogenetic analysis using the stn genes of Salmonella isolated from the aquatic environment, migratory aquatic birds, and chicken showed a strong association between these isolates. In addition, a higher nucleotide identity percentage was observed between the sequences recovered from migratory aquatic birds and Lake Qarun water samples. CONCLUSION: Salmonella distribution was confirmed through migratory aquatic birds, based on our phylogeny tree analysis, Salmonella considered a likely carrier of zoonotic bacterial pathogens. Furthermore, the close relationship between chicken and fish sequences highlights the scenarios of using chicken manure in fish farms and its public health implications. The presence of Salmonella in different environmental sources spotlights the urgent need to control and break down its epidemiological cycle.

Coinfections of Aeromonas spp., Enterococcus faecalis, and Vibrio alginolyticus isolated from farmed Nile tilapia and African catfish in Egypt, with an emphasis on poor water quality.

The high mortality rate among Nile tilapia (Oreochromis niloticus) and African catfish (Clarias gariepinus) polycultured in earthen ponds in Manzala, Egypt, was investigated. Mortality has been linked to poor water quality parameters accompanied with bacterial infections. Moribund farmed fishes exhibited general septicemic signs. Fish from both species (45 each) were sampled and analyzed bacteriologically. Vibrio alginolyticus (32.3%), Enterococcus faecalis (29.4%), Aeromonas caviae (23.5%), and A. veronii (14.7%) were isolated from moribund fishes using selective media and further identified by biochemical tests. 16S rRNA gene sequencing and phylogenetic analysis confirmed the identity of these isolates. Experimental infection of O. niloticus with different bacterial isolates resulted in clinical signs of hemorrhagic septicemia and mortality rates of 80%, 60%, 40%, and 30%, respectively, for E. faecalis, A. veronii, V. alginolyticus, and A. caviae. Water parameter analysis revealed marked divergence from typical values. In addition, different bacterial isolates (including Staphylococcus sciuri, S. aureus, E. faecalis, A. veronii, A. caviae, and V. alginolyticus) were identified and isolated from water samples. BLAST analysis of water bacterial isolates displayed a 100% similarity score with relevant fish isolates, indicating that the water was the likely source of infections. Histopathological examination revealed signs of bacterial infection in both fish species. In addition, common circulatory and degenerative changes with melanophore aggregation, and lymphocytic depletion in hematopoietic organs were recorded.

Isolation and characterization of E. coli strains causing intramammary infections from dairy animals and wild birds.

The study was conducted to estimate the prevalence of Escherichia coli (E. coli) in sub-clinically mastitic (SCM) animals, and in wild and migratory birds which may act as reservoir disseminating such pathogen. Farm hygiene, management and milking procedures were listed through a questionnaire. Thirty lactating cows and 15 lactating buffaloes from five small-scale dairy farms were randomly selected and screened for subclinical mastitis (SCM) using California Mastitis Test (CMT) and somatic cell count (SCC). In addition, 80 teat skin swabs, 5 drinking water samples and 38 wild and migratory bird faecal matter were also collected. All samples were processed for E. coli isolation by culturing on Levine's Eosin Methylene Blue (L-EMB) agar, followed by purification and biochemical identification. Positive samples were subjected to molecular identification and serotyping. In addition, the presence of extended-spectrum beta-lactamase (ESBL) and carbapenemase-producing E. coli have been reported by antimicrobial sensitivity testing. Escherichia coli were isolated from 7.7%, 50% and 50% of the positive CMT cows' quarters, cows' composite and buffaloes' composite milk samples, respectively. In addition, 14% of cows' teats, 20% of water samples, 70% of faecal matter from wild bird, and 33.3% of faecal matter from migratory waterfowls were carrying E. coli. Serotyping, antibiotic-resistant pattern and phylogenetic analysis have pointed the bearable implication of milking hygiene and wild birds in disseminating E. coli strains causing intramammary infections.

Evidence of colistin resistance genes (mcr-1 and mcr-2) in wild birds and its public health implication in Egypt.

Background: Antimicrobial resistance has become one of the most severe global threats to human and veterinary Medicine. colistin is an effective therapeutic agent against multi-drug-resistant pathogens. However, the discovery of transferable plasmids that confer resistance to colistin (mcr-1) has led to challenges in medical science. This study describes the role of wild birds in the harbouring and environmental spread of colistin-resistant bacteria, which could pose a potential hazard to human and animal health. Methods: In total, 140 faecal samples from wild birds (migratory and resident birds) were tested. Twenty surface water samples were collected from the area in which wild bird trapping was conducted, and 50 human stool samples were collected from individuals residing near the surface water sources and farm buildings. Isolation and identification of Enterobacteriaceae and Pseudomonas aeruginosa from the different samples were performed using conventional culture techniques and biochemical identification. PCR amplification of the mcr genes was performed in all positive isolates. Sequencing of mcr-1 genes from three randomly selected E. coli carrying mcr-1 isolates; wild birds, water and humans was performed. Result: The bacteriological examination of the samples showing isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca and P. aeruginosa. The results of multiplex PCR of the mcr genes revealed that E. coli was the most prevalent gram-negative bacterium harbouring the mcr genes, whereas a low prevalence was observed for K. pneumoniae. The prevalence of mcr-1 in resident birds, migratory birds, water sources and humans were 10.4, 20,16.6 and 9.6% while the prevalence of mcr-2 were 1.4, 3.6, 11.1 and 9.6%, respectively. Sequencing of the mcr-1 gene from the three E. coli carrying mcr-1 isolates indicated a possible correlation between the wild bird and surface water isolates. Conclusion: The detection of mcr-1-positive bacteria in wild birds in Egypt indicates the possible environmental dissemination of this gene through bird activity. The impact of the interaction between domestic and wild animals on public health cannot be overlooked.