Surveillance of Escherichia coli in different types of chicken and duck hatcheries: one health outlook.

Escherichia coli is an important zoonotic bacterium that significantly impacts one health concept. E. coli is normally detected in the gut of warm-blooded animals, but some serotypes can cause diseases in humans and animals. Moreover, it can continue for a long time in different environments, replicate in water, and survive outside different hosts. In this study, 171 samples collected from 10 different types of poultry hatcheries (automatic, semiautomatic, and manual "traditional" types) were examined for the prevalence of E. coli. PCR was applied to verify the E. coli isolates via 16S rRNA gene-specific primers. From the gathered samples, 62 E. coli isolates were recovered (36.3%). The highest prevalence was met with the manual "traditional" hatcheries (57.1%) with no significance difference (P = 0.243) in the 3 types of hatcheries. The incidence of E. coli varied significantly in different tested avian types and breeds. The prevalence was 35.7% in duck hatcheries and 37% in chicken hatcheries, with significant differences between breeds of both species (P = 0.024 and 0.001, respectively). The identification of zoonotic E. coli serotypes in this study is concerning, highlighting the need for collaborative efforts across various sectors, including social, environmental, and governance, to promote the adoption of the one health principle in the chicken business. Periodical surveillance, biosecurity measures at the hatcheries and farm levels, and boosting the immunity of birds were recommended to limit the risk of E. coli spread from avian sources to humans.

Virulence, antimicrobial resistance and phylogenetic analysis of zoonotic walking pneumonia Mycoplasma arginini in the one-humped camel (Camelus dromedarius).

In the scientific literature, a small amount of information is found concerning mycoplasmosis in camel species. A variety of pathogens could be causative agents for pneumonia, but walking pneumonia is mostly caused by Mycoplasma with slow development and mild symptoms. The aim of this study was to identify mycoplasmas from camels (Camelus dromedarius) and extending the arsenal of factors implicated in pathogenicity of M. arginini to shed light on the current knowledge gap. 460 lung samples (pneumonic; n=210 and apparently healthy; n=250) were randomly collected from the one-humped camels (C. domedarius) that have been imported from Sudan and slaughtered at Cairo Slaughterhouse. 48 out of 210 isolates (22.9%) recovered from the pneumonic lungs were recorded as M. arginini. Positive PCR results were obtained for all 48 isolates. On the other hand, infection with the organism was not detected in the apparently healthy lungs. Hemolysis and hydrogen sulphide (H2S) production, a compound that has previously not been identified as a virulence factor in M. arginini, was evident in 100% of the isolates. The 48 M. arginini isolates were weak in their ability to form biofilm on polystyrene surfaces. All isolates were 100% susceptible to florfenicol and streptomycin and 100% resistant to ciprofloxacin. Resistance to lincomycin, spiromycin, tylosin, doxacyclin and erythromycin was observed at different frequencies. 13 different combinations of antibiotics representing one to four classes were evident with the Macrolide erythromycin being the most represented. It also should be noted that the ciprofloxacin, doxacyclin, lincomycin, erythromycin combination was the most noted in 21/48 isolates. Surprisingly, none of the virulence genes (vsp, uvrC and gapA) and quinolone resistance genes (parC and gyrA) were detected by PCR.

Enterotoxigenicity and Antibiotic Resistance of Coagulase-Negative Staphylococci Isolated from Raw Buffalo and Cow Milk.

Staphylococcal food poisoning is considered to be one of the most common foodborne illnesses worldwide. Because milk is rich in nutrients and its neutral pH, it leads to the growth of various bacteria. To date, the correlation between enterotoxigenic potential in Staphylococcus species and antimicrobial resistance (AMR), using bioinformatics analysis in buffalo and cow raw milk and the possible health risks from these bacteria, has not been examined in Egypt. A total of 42 Staphylococcus isolates representing 12 coagulase-positive staphylococci (Staphylococcus aureus and Staphylococcus intermedius) and 30 coagulase-negative staphylococci (Staphylococcus capitis, Staphylococcus xylosus, Staphylococcus carnosus, Staphylococcus saccharolyticus, and Staphylococcus auricularis) were isolated. An assay of the antimicrobial resistance phenotypes indicated low resistance against vancomycin (9.5%). The blaZ gene was associated with penicillin G and methicillin resistance and not with sulbactam + ampicillin. The presence of the gene ermB presented the correlation with erythromycin resistance and tetK with tetracycline resistance (correlation index: 0.57 and 0.49, respectively), despite the absence of the same behavior for ermC and tetM, respectively. Interestingly, the gene mecA was not correlated with resistance to methicillin or any other ß-lactam. Correlation showed that slime-producing isolates had more resistance to antibiotics than those of nonslime producers. The multiple correlations between antibiotic resistance phenotypes and resistance genes indicate a complex nature of resistance in Staphylococcus species. The antimicrobial resistance could potentially spread to the community and thus, the resistance of Staphylococcus species to various antibiotics does not depend only on the use of a single antimicrobial, but also extends to other unrelated classes of antimicrobials.

Phoenix dactylifera, mentha piperita and montanide™ ISA-201 as immunological adjuvants in a chicken model.

This study evaluated plant-based immune-adjuvants from crude extracts of Phoenix dactylifera and Mentha piperita as promising adjuvants for vaccines because of the limited side effects associated with plant extracts. In addition, Montanide™ ISA 201 previously used in vaccines in cattle. Eight different infectious coryza (IC) vaccines were prepared from three serovars [A (W strain and local strain), C (Modesto strain) and B (0222 strain)] with eight Avibacterium paragallinarum vaccines adjuvants formulae using liquid paraffin, Montanide™ ISA 71, Montanide™ ISA 201, and Montanide™ Gel adjuvants, P. dactylifera and M. piperita as immune-stimulants at a concentration of 1 mg and 2 mg incorporated with or without liquid paraffin oil as an adjuvant. These vaccines were applied in a chicken model. After a single immunization, the eight vaccine formulations were evaluated using the ELISA and Microplate agglutination test. Evidence of protection in the immunized birds was based on the results after challenge and bacterial isolation. The incorporation of the crude aqueous extract of P. dactylifera or M. piperita at a concentration of 2 mg in a liquid paraffin oil adjuvanted IC vaccine could be employed as an efficient adjuvant for chicken to IC vaccine to enhance immune responses. Also,Montanide™ ISA 201 may be the best adjuvant to be used to enhance the protective response against Av. paragallinarum. Our results confirm that aqueous extracts of M. piperita leaves and P. dactylifera fruit have immunomodulatory potentials in vivo and elevated serum antibodies against Av. Paragallinarum.

Vancomycin and florfenicol resistant Enterococcus faecalis and Enterococcus faecium isolated from human urine in an Egyptian urban-rural community.

Multidrug resistance is one of the top three threats to global public health. Understanding resistance of bacteria is important to help decrease resistance and improve the development of novel antimicrobial agents or other alternative tools to combat public health challenges. Thus, the goal of this study was to investigate the vancomycin and florfenicol resistance genes of five E. faecalis and 15 E. faecium isolated from patients with urinary tract infections. There were 20 Enterococcus obtained from the library collection of randomly selected private hospitals located in the city of El Qanater El Khayreya; these samples were isolated during 2017. Samples were evaluated for their phenotypic characterization of virulence factors, antimicrobial resistance and PCR was conducted to detect the prescence of the vancomycin vanABC and florfenicol resistance genes encoding the catAB, fexAB and cfu. There were six different antibiotic resistance profiles observed. The 20 isolates showed resistance to clindamycin, oxytetracycline and gentamycin. Resistance was evident to ciprofloxacin, norfloxacin and florfenicol in the absence of the cfr gene in all of the 20 Enterococcus isolates. In addition, all isolates produced biofilms and were classified as extensive drug resistant. MARindices of the isolates were >0.6. The MARindex of human isolates of enterococci suggest these pathogens originate from a high-risk source of contamination where antibiotics are often used. This information highlights a possible public health concern to the Egyptian community. The results also suggest the emergence of a linezolid sensitive-vancomycin resistant E. faecium and E. faecalis in the absence of the cfr gene.

Prevalence, Pathogenicity, Virulence, Antibiotic Resistance, and Phylogenetic Analysis of Biofilm-Producing Listeria monocytogenes Isolated from Different Ecological Niches in Egypt: Food, Humans, Animals, and Environment.

Serious outbreaks of foodborne disease have been caused by Listeria monocytogenes found in retail delicatessens and the severity of disease is significant, with high hospitalization and mortality rates. Little is understood about the formidable public health threat of L. monocytogenes in all four niches, humans, animals, food, and environment, in Egypt. This study analyzed the presence of L. monocytogenes collected from the four environmental niches and bioinformatics analysis was implemented to analyze and compare the data. PCR was used to detect virulence genes encoded by pathogenicity island (LIPI-1). prfA amino acid substation that causes constitutive expression of virulence was common in 77.7% of isolates. BLAST analysis did not match other isolates in the NCBI database, suggesting this may be a characteristic of the region associated with these isolates. A second group included the NH1 isolate originating in China, and BLAST analysis showed this prfA allele was shared with isolates from other global locations, such as Europe and North America. Identification of possible links and transmission pathways between the four niches helps to decrease the risk of disease in humans, to take more specific control measures in the context of disease prevention, to limit economic losses associated with food recalls, and highlights the need for treatment options.

Pseudomonas species isolated from camel meat: quorum sensing-dependent virulence, biofilm formation and antibiotic resistance.

Aim: This research pioneers the process of obtaining information concerning the distribution and existence of seven ESBL genes linked to Pseudomonas, three virulence and five quorum sensing separated from 100 camel meat samples using PCR. Materials & methods: The Vitek system was used to identify Pseudomonas species. Phenotypic antibiotic resistance of 16 antibiotics was tested by disc diffusion. Quantification of pyocyanin, elastase, alkaline protease, biofilm and Vero cell cytotoxicity was also implemented. Results: The total number of Pseudomonas species isolated from camel meat was 10/100 identified as Pseudomonas aeruginosa 8/10, Pseudomonas fluorescens 2/10. The isolates were multidrug resistant and were resistant to four to eight antibiotics representing four to six classes. The 15 genes exhibited a huge diversity in their association. Conclusion: The results indicated that camel meat is an unpropitious hotbed for Pseudomonas species of clinical significance.

Poultry as a vector for emerging multidrug resistant Enterococcus spp.: First report of vancomycin (van) and the chloramphenicol-florfenicol (cat-fex-cfr) resistance genes from pigeon and duck faeces.

Although commonly regarded as human and animal intestinal tract commensals, Enterococcus spp. have emerged as important nosocomial pathogens due to their intrinsic or acquired resistance to a number of antibiotics. Poultry has been suggested to be a reservoir for antibiotic resistance that may aggravate the problem of transmission of enterococci infections. Between January and December 2016, 106 Enterococcus spp. were isolated from a total of three poultry species. The collection included isolates recovered from chickens (n = 30), ducks (n = 35) and pigeons (n = 41). All enterococci isolates were screened for their ability to form biofilm. The antibiotic susceptibility was determined against 13 antibiotics using the disc diffusion method. The presence of the eight resistance genes, vanA, vanB, vanC, catA, catB, fexA, fexB and cfr was determined by PCR. All 106 isolates were resistant to clindamycin, whereas majority of isolates (>90%) were resistant to erythromycin, oxytetracycline, doxycycline, gentamycin, ciprofloxacin, norfloxacin, and vancomycin. All isolates produced biofilms and were classified as extensive drug-resistant. MARindices for all isolates was determined to be > 0.8, indicating that they have been recovered from high risk contamination sources. The cfr resistance gene was not detected in any of the 106 enterococci isolates, whereas the chloramphenicol resistance genes catA and catB were found in 18.9% (20/106) of the isolates. Interestengly, fexA 11.9% (15/106), fexB 8.7% (11/106), vanA 18.9% (20/106), vanB 25.5% (27/106), and vanC 33% (35/106) genes were also determined in our study. The present study highlights the emergence of a linezolid sensitive-vancomycin resistant enterococci, which lacks the cfr gene reporting also for the first time the detection of van, fex and cat -genes in Enterococcus species recovered from chickens, ducks and pigeons in Egypt suggesting that poultry species could be potential vectors for transmission of multidrug resistant enterococci posing a public health risk.

Genetic Diversity Among Candida albicans Isolated from Humans and Cattle with Respiratory Distress in Egypt.

As human populaces develop, they are progressively squeezed into higher living densities. The same is true for horticulture and animals expected to bolster these communities. Despite the high potential for zoonotic transmission, connections among humans and cattle have been understudied; however, Candida albicans remains the most important medical mycosis. The genesis of the mycobiome can vary, and interactions between humans and cattle are progressively being perceived as a key interface for disease transmission. αINT1 is a unique gene from Candida albicans; hence, it has been used for detection as well as intraspecific and interspecific phylogenetic analysis of C. albicans collected from human patients and cattle with pulmonary distress in urban-rural populations. A total of 1,921 specimens were examined by direct microscopy and culture to recover yeast associated with human infection. Identification was performed by micromorphology using an API 20C AUX system. The fungal species identified in bovine nasal specimens were Alternaria species (15%), Penicillium species, and C. albicans (6.7%). Other fungal species, such as Aspergillus niger, Torulopsis species, Mucor species (5%), Aspergillus flavus, Fusarium species, Trichosporon species (3.3%), C. rugosa, C. tropical, and Saccharomyces species (1.7%), were also isolated. In human sputum specimens, C. albicans (20%) and C. parapsilosis (2.7%) were the only reported yeast species in our samples. The four identified C. albicans species (two human and two cattle) were subjected to αINT1 gene sequence analysis, which confirmed major phylogenetic relationships among human and cattle isolates. This finding highlights the public health importance of bovines as a potential source for C. albicans zoonotic transmission to humans in an urban-rural community. Additionally, the close relationship between circulating C. albicans strains recorded in Egypt and the United States indicates the possible cross-species transmission of C. albicans between imported foreign and native cattle breeds.

Poultry and beef meat as potential seedbeds for antimicrobial resistant enterotoxigenic Bacillus species: a materializing epidemiological and potential severe health hazard.

Although Bacillus cereus is of particular concern in food safety and public health, the role of other Bacillus species was overlooked. Therefore, we investigated the presence of eight enterotoxigenic genes, a hemolytic gene and phenotypic antibiotic resistance profiles of Bacillus species in retail meat samples. From 255 samples, 124 Bacillus isolates were recovered, 27 belonged to B. cereus and 97 were non-B. cereus species. Interestingly, the non-B. cereus isolates carried the virulence genes and exhibited phenotypic virulence characteristics as the B. cereus. However, correlation matrix analysis revealed the B. cereus group positively correlates with the presence of the genes hblA, hblC, and plc, and the detection of hemolysis (p < 0.05), while the other Bacillus sp. groups are negatively correlated. Tests for antimicrobial resistance against ten antibiotics revealed extensive drug and multi-drug resistant isolates. Statistical analyses didn't support a correlation of antibiotic resistance to tested virulence factors suggesting independence of these phenotypic markers and virulence genes. Of special interest was the isolation of Paenibacillus alvei and Geobacillus stearothermophilus from the imported meat samples being the first recorded. The isolation of non-B. cereus species carrying enterotoxigenic genes in meat within Egypt, suggests their impact on food safety and public health and should therefore not be minimised, posing an area that requires further research.

Urinary tract infection attributed to Escherichia coli isolated from participants attending an unorganized gathering.

AIM: Participants in an unorganized gathering are potential hosts of diseases, bringing diseases from around the world to be introduced to a large at-risk population. Therefore, we investigated the gene repertoire in 29 Escherichia coli strains linked to urinary tract infection isolated from patients transferred to the hospital after attending an unorganized gathering in Cairo. MATERIALS & METHODS: Virulence and resistance determinants, phenotypic antibiotic resistance, biofilm formation, their serotypes and phylogenetic relationships were analyzed. RESULTS: The 29 tested serovars were phenotypically virulent, with the prevalence of group B2, and resistant to tetracycline, naldixic acid, ampicillin, trimethoprim, neomycin, oxytetracycline and erythromycin encoding the iss virulent gene. CONCLUSION: A One Health approach is a must to monitor and control E. coli urinary tract infections.

Poultry hatcheries as potential reservoirs for antimicrobial-resistant Escherichia coli: A risk to public health and food safety.

Hatcheries have the power to spread antimicrobial resistant (AMR) pathogens through the poultry value chain because of their central position in the poultry production chain. Currently, no information is available about the presence of AMR Escherichia coli strains and the antibiotic resistance genes (ARGs) they harbor within hatchezries. Therefore, this study aimed to investigate the possible involvement of hatcheries in harboring hemolytic AMR E. coli. Serotyping of the 65 isolated hemolytic E. coli revealed 15 serotypes with the ability to produce moderate biofilms, and shared susceptibility to cephradine and fosfomycin and resistance to spectinomycin. The most common ß-lactam resistance gene was blaTEM, followed by blaOXA-1, blaMOX-like, blaCIT-like, blaSHV and blaFOX. Hierarchical clustering of E. coli isolates based on their phenotypic and genotypic profiles revealed separation of the majority of isolates from hatchlings and the hatchery environments, suggesting that hatchling and environmental isolates may have different origins. The high frequency of ß-lactam resistance genes in AMR E. coli from chick hatchlings indicates that hatcheries may be a reservoir of AMR E. coli and can be a major contributor to the increased environmental burden of ARGs posing an eminent threat to poultry and human health.

An alternative approach for evaluating the phenotypic virulence factors of pathogenic Escherichia coli.

Escherichia coli is a recognized zoonotic food-borne pathogen; however, the use of polymerase chain reaction (PCR) in the underdeveloped countries to differentiate pathogenic from non-pathogenic E. coli is a problematic issue. Our grail was to assess the phenotypic virulence markers motility, hemolysin, congo red agar, embryo lethality assay and serum resistance for pathogenic E. coli (PEC) correlated to PCR tests which is currently used world-wide to evaluate the PEC. The 448 strains of Escherichia coli that were isolated from different sources, were characterized for phenotypic virulence factors such as motility, hemolysin, Congo red binding, Embryo Lethality assay (ELA) and serum resistance, as well as antibiotic susceptibility using disc diffusion method to 23 antibiotics. Results exhibited 100% motility and Congo red binding, 97.1% for hemolysin production and 90.2% in the ELA. As a result, we were able to hypothetically conclude that the aforementioned virulence markers are plain, straightforward, economical, rapid, more dynamic, uncomplicated methodology, duplicatable and cost next to nothing when compared to the molecular PCR. Their implementation in a diagnostic microbiology laboratory for vetting is a rewarding task in the underdeveloped countries. It augments endeavors to minimize the use of PCR in our investigations especially during epidemiological and outbreak investigations of PEC.

Characterization and susceptibility of streptococci and enterococci isolated from Nile tilapia (Oreochromis niloticus) showing septicaemia in aquaculture and wild sites in Egypt.

BACKGROUND: The present investigation was an endeavor into the elucidation of the disease-causing pathogen of streptococcosis in Nile tilapia (Oreochromis niloticus) in Egypt affecting adult fish cultured and wild fish in the Nile river. Fish were obtained from commercial fishermen, collected as part of their routine fishing activities. The researchers observed the routine fishing process and selected fish for use in the study, at the point of purchase from the fisherman. RESULTS: Diseased fish showed exophthalmia with accumulation of purulent and haemorrhagic fluid around eyes, and ventral petechial haemorrhages. The Post mortem examination revealed, abdominal fat haemorrhage, pericarditis and enlargement of the liver, spleen and kidney. Gram-stained smears revealed the presence of Gram-positive cocci, ß-hemolytic, oxidase and catalase negative. Analysis of the 16S rRNA gene confirmed that the 17 tilapia isolates studied were 6/17 Enterococcus faecalis, 2/17 Enterococcus gallinarum, 3/17 Streptococcus pluranimalium, 2/17 Aerococcus viridans, 1/17 isolate of each Streptococcus dysgalactiae, Streptococcus anginosus, Lactococcus garvieae and Granulicetella elegans/Leuconostoc mesenteroides cremoris. It should be noted that there was no mixed infection. Multiple resistance was observed and the most frequent antibiotic combination was penicillin, ampicillin, vancomycin, chloramphenicol, rifampicin, ofloxacin, clindamycin, erythromycin and tetracycline representing eight classes. CONCLUSIONS: Consequently, we concluded that Streptococcus species are an emerging pathogen for Nile tilapia aquaculture in Egypt and to be considered as a new candidate in the warm water fish diseases in Egypt with special reference to L. garvieae, S. dysgalactiae in addition to L. mesenteroides cremoris which was not reported before from tilapia and taking into consideration their zoonotic implications for public health.

Antimicrobial resistance and virulence characterization of Staphylococcus aureus and coagulase-negative staphylococci from imported beef meat.

BACKGROUND: The objectives of this study were to characterize the diversity and magnitude of antimicrobial resistance among Staphylococcus species recovered from imported beef meat sold in the Egyptian market and the potential mechanisms underlying the antimicrobial resistance phenotypes including harboring of resistance genes (mecA, cfr, gyrA, gyrB, and grlA) and biofilm formation. RESULTS: The resistance gene mecA was detected in 50% of methicillin-resistant non-Staphylococcus aureus isolates (4/8). Interestingly, our results showed that: (i) resistance genes mecA, gyrA, gyrB, grlA, and cfr were absent in Staphylococcus hominis and Staphylococcus hemolyticus isolates, although S. hominis was phenotypically resistant to methicillin (MR-non-S. aureus) while S. hemolyticus was resistant to vancomycin only; (ii) S. aureus isolates did not carry the mecA gene (100%) and were phenotypically characterized as methicillin- susceptible S. aureus (MSS); and (iii) the resistance gene mecA was present in one isolate (1/3) of Staphylococcus lugdunensis that was phenotypically characterized as methicillin-susceptible non-S. aureus (MSNSA). CONCLUSIONS: Our findings highlight the potential risk for consumers, in the absence of actionable risk management information systems, of imported foods and advice a strict implementation of international standards by different venues such as CODEX to avoid the increase in prevalence of coagulase positive and coagulase negative Staphylococcus isolates and their antibiotic resistance genes in imported beef meat at the Egyptian market.

Prevalence of the Antibiotic Resistance Genes in Coagulase-Positive-and Negative-Staphylococcus in Chicken Meat Retailed to Consumers.

The use of antibiotics in farm management (growing crops and raising animals) has become a major area of concern. Its implications is the consequent emergence of antibiotic resistant bacteria (ARB) and accordingly their access into the human food chain with passage of antibiotic resistance genes (ARG) to the normal human intestinal microbiota and hence to other pathogenic bacteria causative human disease. Therefore, we pursued in this study to unravel the frequency and the quinolone resistance determining region, mecA and cfr genes of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), methicillin-resistant coagulase-negative staphylococci (MRCNS) and methicillin-susceptible coagulase-negative staphylococci (MSCNS) isolated from the retail trade of ready-to-eat raw chicken meat samples collected during 1 year and sold across the Great Cairo area. The 50 Staphylococcus isolated from retail raw chicken meat were analyzed for their antibiotic resistance phenotypic profile on 12 antibiotics (penicillin, oxacillin, methicillin, ampicillin-sulbactam, erythromycin, tetracycline, clindamycin, gentamicin, ciprofloxacin, chloramphenicol, sulfamethoxazole-trimethoprim, and vancomycin) and their endorsement of the quinolone resistance determining region, mecA and cfr genes. The isolation results revealed 50 isolates, CPS (14) and CNS (36), representing ten species (S. aureus, S. hyicus, S. epidermedius, S. lugdunensis, S. haemolyticus, S. hominus, S. schleiferi, S. cohnii, S. intermedius, and S. lentus). Twenty seven isolates were methicillin-resistant. Out of the characterized 50 staphylococcal isolates, three were MRSA but only 2/3 carried the mecA gene. The ARG that bestows resistance to quinolones, ß-lactams, macrolides, lincosamides, and streptogramin B [MLS(B)] in MRSA and MR-CNS were perceived. According to the available literature, the present investigation was a unique endeavor into the identification of the quinolone-resistance-determining-regions, the identification of MRSA and MR-CNS from retail chicken meat in Egypt. In addition, these isolates might indicate the promulgation of methicillin, oxacillin and vancomycin resistance in the community and imply food safety hazards.

Determination of virulence and antibiotic resistance pattern of biofilm producing Listeria species isolated from retail raw milk.

BACKGROUND: One of the foodborne pathogens is Listeria monocytogenes, which causes serious invasive illness in elderly and immunocompromised patients, pregnant women, newborns and infants ranking second after salmonellosis because of its high case fatality rate. Listerial cow mastitis marked by abnormal milk, increased cell counts and reduced production has not been reported. Therefore, apparently healthy cows can be reservoirs of L. monocytogenes. A number of 203 udder milk samples from apparently healthy animals (buffalo, n = 100; cow, n = 103) were collected and tested for Listeria. Isolated colonies on the PALCAM agar were Listeria species confirmed according to their biochemical and the Christie-Atkins-Munch-Petersen (CAMP) reactions. The Listeria species pathogenicity of was tested by phosphatidylinositol-specific phospholipase C, DL-alanine-ß-naphthylamide HCl, Dalanine-p-nitroanilide tests, chick embryo, mice inoculation tests, Vero cell cytotoxicity and biofilm formation. The virulence-associated genes, hlyA, plcB, actA and iap associated with Listeria were molecularly assayed. RESULTS: The 17 isolated Listeria spp. represented a prevalence rate of 8.4 %. Of these 3 (1.4 %), 2 (1 %), 5 (2.5 %), 4 (2 %) and 3 (1.5 %) were confirmed as L. monocytogenes, L. innocua, L. welshimeri, L. seelegeri, respectively. While the L. monocytogenes isolate displayed all the four virulence-associated genes, L. seelegeri carried the hlyA gene only. The L. monocytogenes had a strong in vitro affinity to form a biofilm, in particular serotype 4 which is associated with human infections. L. monocytogenes showed resistance for 9/27 antibiotics. CONCLUSIONS: The biofilm forming capability of the Listeria spps. makes them particularly successful in colonizing surfaces within the host thus being responsible for persistence infections and due to their antimicrobial resistant phenotype that this structure confers. In addition, strains belonging to serotypes associated with human infections and characterized by pathogenic potential (serotype 4) are capable to persist within the processing plants forming biofilm and thus being a medical hazard.

Dispersion of the Vancomycin Resistance Genes vanA and vanC of Enterococcus Isolated from Nile Tilapia on Retail Sale: A Public Health Hazard.

Although normally regarded harmless commensals, enterococci may cause a range of different infections in humans, including urinary tract infections, sepsis, and endocarditis. The acquisition of vancomycin resistance by enterococci (VRE) has seriously affected the treatment and infection control of these organisms. VRE are frequently resistant to all antibiotics that are effective treatment for vancomycin-susceptible enterococci, which leaves clinicians treating VRE infections with limited therapeutic options. With VRE emerging as a global threat to public health, we aimed to isolate, identify enterococci species from tilapia and their resistance to van-mediated glycopeptide (vanA and vanC) as well as the presence of enterococcal surface protein (esp) using conventional and molecular methods. The cultural, biochemical (Vitek 2 system) and polymerase chain reaction results revealed eight Enterococcus isolates from the 80 fish samples (10%) to be further identified as E. faecalis (6/8, 75%) and E gallinarum (2/8, 25%). Intraperitoneal injection of healthy Nile tilapia with the eight Enterococcus isolates caused significant morbidity (70%) within 3 days and 100% mortality at 6 days post-injection with general signs of septicemia. All of the eight Enterococcus isolates were found to be resistant to tetracycline. The 6/6 E. faecalis isolates were susceptible for penicillin, nitrofurantoin, gentamicin, and streptomycin. On the other hand 5/6 were susceptible for ampicillin, vancomycin, chloramphenicol, and ciprofloxacin. The two isolates of E. gallinarum were sensitive to rifampicin and ciprofloxacin and resistant to vancomycin, chloramphenicol, and erythromycin. Molecular characterization proved that they all presented the prototypic vanC element. On the whole, one of the two vancomycin resistance gene was present in 3/8 of the enterococci isolates, while the esp virulence gene was present in 1/8 of the enterococci isolates. The results in this study emphasize the potential role that aquatic environments are correlated to proximity to anthropogenic activities in determining the antimicrobial resistance patterns of Enterococcus spp. recovered from fish in the river Nile in Giza, Elmounib, Egypt as a continuation of our larger study on the reservoirs of antibiotic resistance in the environment.

Antimicrobial Resistance, Biofilm Formation and mecA Characterization of Methicillin-Susceptible S. aureus and Non-S. aureus of Beef Meat Origin in Egypt.

Methicillin-resistant Staphylococcus aureus (MRSA) have been found in various farm animal species throughout the world. Yet, methicillin-susceptible S. aureus (MSSA), methicillin-susceptible non-S. aureus (MS-NSA), and methicillin-resistant non-S. aureus (MR-NSA) were not investigated. Therefore, we persued to determine the diversity in their phenotypic virulence assay, phenotypic antimicrobial resistance profile and molecular characterization in one of the food chains in Egypt. Samples were collected during 2013 from beef meat at retail. Twenty seven isolates comprising five species (S. hyicus, S. aureus, S. schleiferi subsp. coagulans, S. intermedius, and S. lentus) were characterized for their antibiotic resistance phenotypic profile and antibiotic resistance genes (mecA, cfr, gyrA, gyrB, and grlA). Out of the 27 Staphylococcus isolates only one isolate was resistant to the 12 antibiotics representing nine classes. Raw beef meat sold across the Great Cairo zone, contains 66.7% of MRS, with highest prevalence was reported in S. aureus (66.7%), while the MRS non-S. aureus strains constituted 66.7% from which S. hyicus (60%), S. intermedius (33.3%), S. schleiferi subsp. coagulans (100%), and S. lentus (100%) were MRS. Seven S. aureus, six S. hyicus, four S. schleiferi subsp. coagulans, three S. intermedius, and one S. lentus isolates although being resistant to oxacillin yet, 11/27 (40.7%) carried the mecA gene. At the same time, the cfr gene was present in 2 of the nine S. aureus isolates, and totally undetectable in S. hyicus, S. schleiferi subsp. coagulans, S. intermedius, and S. lentus. Although, global researches largely focused into MRSA and MR-NSA in animals on pigs, the analysis of our results stipulates, that buffaloes and cattle could be MRSA dispersers and that this theme is not specific to pigs. Detection of MSSA virulence determinants is a must, as although oxacillin resistance may be absent yet, the MSSA may carry the virulence determinants which could be a source of perilous S. aureus for the human community.

Prevalence and antimicrobial resistance profile of Staphylococcus species in chicken and beef raw meat in Egypt.

Coagulase-positive (CPS) and coagulase-negative (CNS) staphylococci cause staphylococcal food poisoning. Recently, CPS and CNS have received increasing attention due to their potential role in the dissemination of antibiotic resistance markers. The present study aimed to evaluate CPS and CNS species distribution and their antibiotic resistance profile isolated from chicken and beef meat. Fifty fresh, uncooked chicken parts and 50 beef meat cuts (local n=27; imported n=23) were used. One hundred staphylococcal isolates belonging to 11 species were isolated and identified from chicken (n=50) and beef (n=50) raw meat samples. Staphylococcus hyicus (26/100), lugdunensis (18/100), aureus (15/100) and epidermidis (14/100) were dominant. S. aureus was 100% resistant to penicillin and sulfamethoxazole/trimethoprim. Vancomycin-resistant S. aureus showed intermediate resistance (51%), which might indicate the dissemination of vancomycin resistance in the community and imply food safety hazards. The percentage of resistance to ß-lactams was variable, with the highest resistance being to penicillin (94%) and lowest to ampicillin-sulbactam (22%). Antimicrobial resistance was mainly against penicillin (94%), clindamycin (90%) and sulfamethoxazole/trimethoprim (82%). The results indicate that chicken and beef raw meat are an important source of antibiotic-resistant CPS and CNS.