Dissemination, virulence characteristic, antibiotic resistance determinants of emerging linezolid and vancomycin-resistant Enterococcus spp. in fish and crustacean.

Enterococci are emerging nosocomial pathogens. Their widespread distribution causes them to be food contaminants. Furthermore, Enterococci can colonize various ecological niches and diffuse into the food chain via contaminated animals and foods because of their remarkable tolerance to unfavorable environmental circumstances. Due to their potential dissemination to humans, antimicrobial-resistant Enterococci in fish are a worldwide health issue. This study characterized AMR, ARGs, VAGs, gelatinase activity, and biofilm formation in Enterococcus spp. recovered from fish and seafood and evaluated potential correlations. 54 Enterococcus spp. strains(32.73 %)were isolated from 165 samples (75 Oreochromis niloticus, 30 Argyrosomus regius, and 60 Shrimp), comprising 30 Enterococcus faecalis (55.6 %) and 24 Enterococcus faecium (44.4 %) with total 32.73 % (54/165), The maximum prevalence rate of Enterococcus spp. was observed in Nile tilapia (34/54; 63 %), followed by shrimp (14/54; 25.9 %) and Argyrosomus regius (6/54; 11.1 %). The maximum prevalence rate of E. faecalis was observed in Nile tilapia (22/30; 73.3 %), followed by shrimp (8/30; 26.7 %) with significant differences. The prevalence rate of E. faecium was observed in Nile tilapia (12/24; 50 %), followed by shrimp (6/24,25 %). E. faecium is only isolated from Argyrosomus regius (6/24,25 %). Isolates exhibited high resistance against both tetracycline (90.7 %) and erythromycin(88.9 %), followed by gentamycin (77.8 %), ciprofloxacin (74.1 %), levofloxacin (72.2 %), penicillin (44.4 %), vancomycin (37 %), and linezolid (20.4 %). 50 strains (92.6 %) exhibited resistance to more than two antibiotics, 5 strains (10 %) were XDR, and the remaining 45 strains (90 %) were classified as MDR. 92.6 % of the isolates had MARindices >0.2, indicating they originated in settings with a high risk of contamination. Additionally, ten ARGs were identified, with tet(M) 92.6 %, followed by erm(B) (88.9 %), aac(6')-Ie-aph(2″)-Ia(77.8 %), tet(K) (75.9 %), gyrA (74.1 %), blaZ (48.1 %), vanA (37 %), vanB (31.5 %), optrA (20.4 %), and catA(3.7 %). Biofilm formation and gelatinase activity were observed in 85.2 %, and 61.1 % of the isolates, respectively. A total of 11 VAGs were detected, with gelE as the most prevalent (83.3 %) followed by agg(79.6 %), pil (74.1 %), both sprE and asa1 (72.2 %), hyl (70.4 %), eps(68.5 %), EF3314 (57.4 %), ace (50 %), and cylA (35.2 %) with no detection of cylB. In conclusion, the emergence of linezolid-resistant -vancomycin-resistant enterococci recovered from Egyptian fish and shrimp, suggests that fish and seafood might participate a fundamental part in the emergence of antimicrobial resistance among humans.

Assessment of some toxic elements (Co, Cr, Mn, Se, and As) in muscle, offal, hair, and blood of camels (Camelus dromedaries) and their risk assessment.

Background: Camel meat tainted with heavy metals or trace elements may pose a health risk to consumers. Heavy metal contamination poses a severe danger due to both their toxicity and bioaccumulation in the food chain. Aim: To estimate the residual levels of heavy metals (Co, Cr, Mn, Se, and As) in muscle, liver, kidney, hair, and serum of three camel breeds (Magaheem, Maghateer, and Wadha) collected from Al-Omran abattoir, Al-Ahsa, Saudi Arabia. Methods: A total of 225 tissue samples (muscles, liver, kidney, serum, and hair) were taken and analyzed using an Atomic Absorption Spectrophotometer. Health risk assessment was assessed using the guidelines set by the US Environmental Protection Agency. Results: Camel breed significantly (p < 0.05) influences Co, Cr, Mn, and Se accumulation and distribution in organs and muscle; however, arsenic accumulation was not significantly affected (p < 0.05) by camel breeds. The highest values of Co, Cr, Se, and Mn in all examined samples were detected in the liver samples of Maghateer and Magaheem breeds. Furthermore, significant strong positive correlation between serum and liver cobalt, chromium, manganese, and arsenic. The estimated daily intake owing to camel meat consumption was less than the tolerated daily intake. Conclusion: Heavy metals were distributed among different breeds of camel. Trace elements (Pb and Cd) in meat and offal were below the international maximum permissible limit. The correlation between samples reflects the role of hair as a good tool for the identification of heavy metal pollution.

Prevalence and antibiogram of shigatoxinproducing E. coli in camel meat and offal.

Background: Camels are important animals in Egypt and other Arab countries on the basis of their economic value and ethnic culture. Escherichia coli is implicated in several gastrointestinal infections and outbreaks worldwide, especially in developing countries. It causes infections that might lead to death. Numerous biological activities, such as antioxidative, antibacterial, anti-diabetic, anti-mutagenic, anti-inflammatory, neuroprotective, and diuretic, are associated with coriander and coriander essential oils. Aim: This work targeted investigation of the prevalence, antibiogram, and occurrence of virulence genes of E. coli in camel meat liver and kidney. Besides, the anti-E. coli activity of coriander oil was further examined. Methods: Camel meat, liver, and kidneys were collected from local markets in Egypt. Isolation and identification of E. coli were performed. The antimicrobial susceptibility of the obtained E. coli isolates was screened using the disk diffusion assay. To detect the presence of virulence-associated genes (stx1, stx2, eaeA, and hylA gens), polymerase chain reaction was used. An experimental trial was done to investigate the anti-E. coli activity of coriander oil. Results: The obtained results revealed isolation of the following E. coli pathotypes: O17:H18, O128:H2, O119:H6, O103:H4, O145:H-, and O121:H7. The recovered E. coli isolates practiced multidrug resistance profiling with higher resistance toward Erythromycin, Nalidixic Acid, Clindamycin, and Ampicillin. However, the isolates were sensitive to Meropenem and cefoxitin. The recovered isolates had expressed different virulence attributes. Coriander oil of 2% could significantly reduce E. coli O128 count in camel meat by 65%. Conclusion: Therefore, strict hygienic measures are highly recommended during the processing of camel meat. The use of coriander oil during camel meat processing is highly recommended to reduce E. coli count.

Cross-sectional analysis of risk factors associated with Mugil cephalus in retail fish markets concerning methicillin-resistant Staphylococcus aureus and Aeromonas hydrophila.

Introduction: Aeromonas hydrophila and methicillin-resistant Staphylococcus aureus (MRSA) are potent bacterial pathogens posing major hazards to human health via consuming fish harboring these pathogens or by cross-contamination beyond the contaminated environment. The aim of this study was to determine risk variables associated with the presence of certain pathogenic bacteria from Mugil cephalus fish in retail markets in Egypt. The virulence genes of A. hydrophila and S. aureus were also studied. Furthermore, the antibiotic sensitivity and multidrug resistance of the microorganisms were evaluated. Methods: In a cross-sectional investigation, 370 samples were collected from mullet skin and muscle samples, washing water, fish handlers, knives, and chopping boards. Furthermore, fish handlers' public health implications were assessed via their response to a descriptive questionnaire. Results: S. aureus and Aeromonas species dominated the investigated samples with percentages of 26.76% and 30.81%, respectively. Furthermore, A. hydrophila and MRSA were the predominant recovered bacterial pathogens among washing water and knives (53.85% and 46.66%, respectively). The virulence markers aerA and hlyA were found in 90.7% and 46.5% of A. hydrophila isolates, respectively. Moreover, the virulence genes nuc and mec were prevalent in 80% and 60% of S. aureus isolates, respectively. Antimicrobial susceptibility results revealed that all A. hydrophila isolates were resistant to amoxicillin and all MRSA isolates were resistant to amoxicillin and ampicillin. Remarkably, multiple drug resistance (MDR) patterns were detected in high proportions in A. hydrophila (88.37%) and MRSA (100%) isolates. The prevalence of Aeromonas spp. and S. aureus had a positive significant correlation with the frequency of handwashing and use of sanitizer in cleaning of instruments. MRSA showed the highest significant prevalence rate in the oldest age category. Conclusion: The pathogenic bacteria recovered in this study were virulent and had a significant correlation with risk factors associated with improper fish handling. Furthermore, a high frequency of MDR was detected in these pathogenic bacteria, posing a significant risk to food safety and public health.

Genomic traits of Aeromonas veronii isolated from slaughtered Danish broilers.

Aeromonas veronii is a Gram-negative opportunistic bacterium found in fish, poultry and humans and has occasionally been associated with disease although not generally considered a poultry pathogen. A. veronii was recently isolated from both healthy and condemned broiler carcasses at a major Danish abattoir. In this study, we did a whole genomic analysis of 24A. veronii strains from the abattoir to determine their potential sources and relatedness as well as their pathogenic potential, antimicrobial resistance determinants and associated mobile elements. No strains were multi-drug resistant, but all strains carried the beta-lactam resistance genes cphA3 and blaOXA-12 without being phenotypically resistant to carbapenems. One strain carried an IncA plasmid with tet(A), tet(B) and tet(E) genes. A phylogenetic tree including public A. veronii sequences showed that our isolates were not clonal but were dispersed around the phylogenetic tree, suggesting a diffuse spread of A. veronii across human, aquatic and poultry samples. Strains carried different virulence factors known to be associated with pathogenesis and severity of disease in animals and humans, e.g. type II (aerolysin, amylases, proteases, and cytotoxic enterotoxin Act) and III secretion systems where the latter has been associated with mortality in hospitalized patients. Although our genomic analysis of A. veronii shows zoonotic potential, epidemiological studies of human gastro-enteritis cases of A. veronii associated with consumption of broiler meat are needed. It remains to be proven if A. veronii is a true poultry pathogen and part of the established microflora in abattoirs and the gut-intestinal microflora of poultry.

Characterization of Escherichia coli and other bacteria isolated from condemned broilers at a Danish abattoir.

Meat inspection is important to ensure food safety and protect public health. Visual inspection of slaughtered carcasses for pathological changes should be supported by bacteriological analysis to determine whether the entire carcass or parts of it should be condemned. The aim of this study was to determine the bacterial species present in different sample types from condemned broiler carcasses. Furthermore, we investigated the genetic characteristics, zoonotic potential, and relatedness of Escherichia coli, the predominant bacterial species isolated from the carcasses. A total of 400 broiler carcasses condemned because of cellulitis (100), scratches (100), hepatitis (100), and healthy control carcasses (100) were selected. Samples of meat, pathological lesion, and bone marrow of each carcass were obtained for microbial analysis. From the analyzed samples, 469 bacterial isolates were recovered with E. coli accounting for 45.8%, followed by Aeromonas spp. (27.9%), in particular A. veronii. The highest rate of bacterial isolation was observed in carcasses condemned with cellulitis, whereas carcasses with hepatitis had the lowest rate of bacterial isolation. Forty-four E. coli isolates originating from different sample types were selected for whole genome sequencing. A clonal relationship was shown between E. coli from different sample types of the same carcass condemned with cellulitis and scratches. A major clade of E. coli was found in carcasses condemned with cellulitis with isolates containing mdf(A), tet(A), and bla TEM-1B genes that confer resistance to macrolides, tetracycline, and ampicillin, respectively. E. coli in this clade all belonged to ST117 and clustered with E. coli isolates previously collected from dead chickens and carcasses condemned due to cellulitis in Denmark, Finland, and the United Kingdom. Bacterial evaluation results of carcasses condemned with cellulitis, scratches (moderate to severe skin lesion), and acute hepatitis confirmed the need for total condemnation of carcasses with these pathological findings. A similar evaluation should be done for carcasses affected with chronic hepatitis, and minor scratches lesions.

The association between meat inspection codes, footpad lesions and thinning of broiler flocks in the Danish broiler production.

The foundation of the condemnation practices in Post-Mortem Inspection (PMI) of poultry should be based on up-to-date scientific evidence about the cause of infection and hence whether the lesions observed are of food safety, animal health or welfare concerns. This study aimed to investigate the association between meat inspection codes, footpad lesions, and thinning of flocks in Danish broiler production. The data set was based on the delivery of chicken flocks to one of the two larger chicken slaughterhouses in Denmark, representing 71 farms, 174 houses, and 4,068 flocks over three years from January 2016 to December 2018. Post-mortem condemnation data of slaughtered chickens recorded and stored in the Danish Quality Assurance System (KIK) database was used in the study. Five potentially causal models were developed to investigate whether there was an association between dermatitis, arthritis, systemic infection, emaciation, mortality and possible explaining factors` (footpad lesion, age at slaughter, scratches, ascites, systemic infection and thinning of the flock). These five ecological logistic regression models were analyzed with the three levels: farm, house, and flock. Data from a total number of 126,137,002 (N) slaughtered chickens recorded in KIK databases were used for modeling and analyses. The prevalence of condemned carcasses was 1.1 % (n = 1,420,812). Overall, 12 individual reasons for condemnation of carcasses were recorded. The most frequently observed reason for condemnation was skin disease (scratches and dermatitis) with a prevalence of 0.5 %. Prevalence of ascites was 0.2 %, discoloration 0.09 %, emaciation 0.09 %, hepatitis 0.09 % and arthritis 0.07 %. In the first model, dermatitis was shown to be positively associated with age at slaughter with an OR = 1.04 (CI95 %: 1.02-1.05), while arthritis was considered an intervening variable. Moreover, there was a small protective effect of thinning of the flock for first and second delivery. There was a positive association between arthritis and age at the time of slaughter with an OR = 1.13 (CI95 %: 1.12-1.15). Systemic infections were associated with scratches with an OR = 24.5 (CI95 %: 16.6-36.3) and footpad lesions with an OR = 1.007 (CI95 %: 1.006-1.008). Further modelling of emaciation and mortality was not considered because of unbalanced groups in the data probably caused by the fact that some condemnation codes were rare. We observed that the most common causal factors of condemnation in the systemic infection models were scratches and footpad lesion, therefore preventing and controlling such lesions could reduce losses. Specific management and environmental etiological factors of the main infections causing condemnation in Danish broilers should be determined.