Detection of Multi-Drug Resistant Food-borne Bacteria in Ready-to-Eat Meat Products in Luxor City, Egypt

A total of five Escherichia coli and eight Salmonella isolates were recovered from ready-to-eat meat samples obtained from different street vendors in Luxor city, Egypt. Bacterial isolates were assayed for antimicrobial susceptibility, its virulence and antimicrobial resistance genes. The total number recovered positive Salmonella spp and E. coli were 8 (6.66%) and 5 (4.16%) respectively. All E. coli isolates were exhibited resistance against streptomycin and cephalothin. While all Salmonella isolates were resistant to nalidixic acid. PCR screening for virulence genes showed that 2 (40%) of the E. coli (O111:H4) serovar were positive for stx1, stx2, and eaeA. While Salmonella enteritidis, typhimurium, and virchow hold invA, hilA and stn genes with percentage of 37.5, 25 and 12.5% respectively. The identified tetracycline resistance gene for E. coli isolates were tetB (60%), tetC (20%) and tetD (20%). The β-lactamase resistance gene blaCTX was identified in 50% of Salmonella isolates represented by S. enteritidis, S. typhimurium and S. Virchow. The blaCMY genes were detected in S. typhimurium and S. infantis (37.5%). These results highlighted the role of ready-to-eat meat as a potential source for multidrug-resistant strains of E. coli and Salmonella. The current results indicate the need for applying hygienic practices in food outlets - especially in street vendors - to reduce the incidence of food-borne bacteria and to prevent future food-borne outbreaks in the studied area

Prevalence of Toxigenic and Methicillin Resistant Staphylococci in Poultry Chain Production

oai:ojs.pkp.sfu.ca:article/1Staphylococci are a worldwide cause of human and animal infection and are considered to be of the most common causes of infections in birds. Enterotoxins produced by some staphylococcal species were recognized as a causative agent of staphylococcal food poisoning (SFP). Only enterotoxins produced by Staphylococcus aureus were as yet well characterized. Much less is known about enterotoxigenic potential of coagulase-negative species of genus Staphylococcus (CNS). It has been reported that enterotoxigenic CNS strains have been associated with human and animal infections and food poisoning. Samples collected from chicken production cycle (un hatched eggs, baby chicks, broilers, chicken meat and table eggs) in Luxor, Egypt were tested to investigate the presence of Staphylococcus species and detection of their enterotoxines genes with more special attention for detection of methicillin resistance gene (mec A). Samples were tested for S. aureus and CNS on the basis of cultural and biochemical properties and confirmed by PCR amplification of 16S rRNA and clfa gene. Results showed that the presence of Staphylococci were 50/150 (33.3%), 14% of the samples were S. aureus (21/150), while, 19.33% were CNS (29/150). mecA gene was detected in 66.7% and 51.7% among S. aureus and CNS respectively. Enterotoxins genes (seb, sec and see) were found in 5 (23.8%) of S. aureus with a percent of (9.5%) for seb and sec and (4.8%) for see, while sec and see were found in 6 (20.6%) of CNS.  With a percent (10.3%) for each. &nbsp

Molecular Characterization of Biofilm Producing Genes in Salmonellae Isolated from Chicken

Salmonella enterica considered one of the most important food-borne pathogen. Biofilm formation considered one of the main problems related to S. enterica, in this study, biofilm formation, colony morphotype, cellulose and curli production genes of 19 Salmonella isolates were tested. The results showed that 85% of isolates produced strong biofilm and 15% of isolates produced moderate biofilm on polystyrene plate with 1/20 diluted TSB. Different colony morphotypes expressed saw, sbam, and rdar morphotype when cultivated on LB containing Congo red for monitoring cellulose and curli production. All S. enterica strains possess adrA, csgD and gcpA genes using PCR. Thus in this study all Salmonella isolates formed biofilm so they give increased tolerance for antimicrobial agents and disinfectant, which results in difficulty in the treatment of diseases and causing many problems in food industry as it becomes a persistent of source of contamination

Surveillance on A/H5N1 virus in domestic poultry and wild birds in Egypt

The endemic H5N1 high pathogenicity avian influenza virus (A/H5N1) in poultry in Egypt continues to cause heavy losses in poultry and poses a significant threat to human health. Here we describe results of A/H5N1 surveillance in domestic poultry in 2009 and wild birds in 2009-2010. Tracheal and cloacal swabs were collected from domestic poultry from 22024 commercial farms, 1435 backyards and 944 live bird markets (LBMs) as well as from 1297 wild birds representing 28 different types of migratory birds. Viral RNA was extracted from a mix of tracheal and cloacal swabs media. Matrix gene of avian influenza type A virus was detected using specific real-time reverse-transcription polymerase chain reaction (RT-qPCR) and positive samples were tested by RT- qPCR for simultaneous detection of the H5 and N1 genes. In this surveillance, A/H5N1 was detected from 0.1% (n = 23/) of examined commercial poultry farms, 10.5% (n = 151) of backyard birds and 11.4% (n = 108) of LBMs but no wild bird tested positive for A/H5N1. The virus was detected from domestic poultry year- round with higher incidence in the warmer months of summer and spring particularly in backyard birds. Outbreaks were recorded mostly in Lower Egypt where 95.7% (n = 22), 68.9% (n = 104) and 52.8% (n = 57) of positive commercial farms, backyards and LBMs were detected, respectively. Higher prevalence (56%, n = 85) was reported in backyards that had mixed chickens and waterfowl together in the same vicinity and LBMs that had waterfowl (76%, n = 82). Our findings indicated broad circulation of the endemic A/H5N1 among poultry in 2009 in Egypt. In addition, the epidemiology of A/H5N1 has changed over time with outbreaks occurring in the warmer months of the year. Backyard waterfowl may play a role as a reservoir and/or source of A/H5N1 particularly in LBMs. The virus has been established in poultry in the Nile Delta where major metropolitan areas, dense human population and poultry stocks are concentrated. Continuous surveillance, tracing the source of live birds in the markets and integration of multifaceted strategies and global collaboration are needed to control the spread of the virus in Egypt

Bacteriological and molecular study of Salmonella species associated with central nervous system manifestation in chicken flocks

Background and Aim: Salmonella species often cause systemic health problems in poultry flocks, sometimes including nervous systems manifestations. This impact of Salmonella has rarely been studied. This study aimed to define an alternative pathogenic pathway for Salmonella spp. invasion of brain tissue in chicken flocks. Brain infection produces neurological manifestations; Salmonella strains isolated from brain tissue showed the presences of two virulence genes. Confirmation of the pathway of isolates from intestinal mucosa through the blood–brain barrier was attained using experimental infections in specific pathogen-free (SPF)-day-old chicks through two routes of inoculation. Materials and Methods: Isolation of Salmonella spp. from five chicken flocks that showed signs of the central nervous system (CNS) effects were isolated. Isolates were characterized by serotyping, and antimicrobial assays. In addition, virulence profiles were described using detection of virulence plasmid spvC, and Salmonella plasmid sopB. A pathogenicity study of isolates in specific pathogen-free (SPF)-day-old chicks through oral and intracerebral administration performed, and experimental infection in SPF embryonated chicken eggs through intra-yolk and intra-allantoic administration was investigated. Supporting histopathology and immunohistopathology against Salmonella antigen in brain tissue were performed for flock and experimental infections. Results: Three serotypes of Salmonella were isolated from the brains of five flocks (two Salmonella Virchow, two Salmonella Kentucky, and one Salmonella Enteritidis isolates). Phage related gene sopB and plasmid-mediated operon spvC were identified in all isolated strains. The Salmonella strains were re-isolated and identified from the brain and internal organs of post-experimental infected chicks. Infected chicks showed nervous manifestations associated with Salmonella infection. The presence of positively stained Salmonella antigen in brain tissues indicates penetration of the blood–brain barrier by the Salmonella species. Conclusion: Our results indicate that some virulent systemic strains of Salmonella spp. can induce CNS manifestations in chicken hosts

A comparative study on the use of real time polymerase chain reaction (RT-PCR) and standard isolation techniques for the detection of Salmonellae in broiler c

This study was carried out to compare between conventional cultural isolation methods and real time polymerase chain reaction (RT-PCR) technique for the detection of Salmonella in broiler chicks. About 120 livers and intestinal contents samples were collected from 1800 day-old imported and local broiler chicks. The incidence of Salmonellae among imported chicks was 11.67% compared to 21.67% among local chicks using conventional cultural isolation methods. Salmonella newport (S. newport) showed the highest incidence rate in imported chicks, while Salmonella enteritidis and Salmonella typhimurium were frequently detected in local chicks. The RT-PCR results for detection of invA gene of Salmonella spp. were 58.33% and 66.67% positive samples in imported and local chicks, respectively. Results have confirmed that RT-PCR technique is rapid, robust, effective and reliable method for detection of Salmonella spp. in broiler chicken when compared to conventional cultural methods. However, RT-PCR should be performed parallel with conventional methods for more accurate detection results of different Salmonellae serovars

Detection of Novel Goose Parvovirus Disease Associated with Short Beak and Dwarfism Syndrome in Commercial Ducks

Derzsy’s disease causes disastrous losses in domestic waterfowl farms. A genetically variant strain of Muscovy duck parvovirus (MDPV) and goose parvovirus (GPV) was named novel goose parvovirus (NGPV), which causes characteristic syndrome in young ducklings. The syndrome was clinically characterized by deformity in beaks and retarded growth, called short beaks and dwarfism syndrome (SBDS). Ten mule and pekin duck farms were investigated for parvovirus in three Egyptian provinces. Despite low recorded mortality rate (20%), morbidity rate was high (70%), but the economic losses were remarkable as a result of retarded growth and low performance. Isolation of NGPV was successful on primary cell culture of embryonated duck liver cells with a clear cytopathic effect. Partial gene sequence of the VP1 gene showed high amino acids identity among isolated strains and close identity with Chinese strains of NGPV, and low identity with classic GPV and MDPV strains. To the best of our knowledge, this can be considered the first record of NGPV infections in Egypt

Phenotypic and Genotypic Characterization of Paratyphoid Salmonellae isolated from Poultry in Delta Area- Egypt

The present work aimed to isolate and characterize Salmonellae from chickens, ducks, quails and turkeys in five Egyptian Governorates. Polymerase chain reaction (PCR) was used for the detection of common virulence genes. A total of 265 flock samples (150 chickens, 60 ducks, 30 quails and 25 turkeys) were collected from Dakahlia, Kafrelsheik, Damietta, Sharkia and Gharbia Governorates. Birds were subjected to either clinical and/or post-mortem examination, in adittion to isolation and identification of salmonellae from internal organs including liver, lung, spleen, caecum and unabsorbed yolk sac. Biochmeical and serological identification of the isolates was done. Twenty eight birds (10.6%) were found positive for Salmonella isolation. The number and percentage of positive chickens, ducks, quails and turkeys were 16 (10.7%), 7 (11.7%), 3 (10%) and 2 (8%), respectively. Salmonella Typhimurium, S. Enteritidis, S. Kentucky, S. Paratyphi A, S. Molade, S. Heidelberg, S. Infantis and S. Apeyeme were isolated from chickens. While S. Enteritidis, S. Typhimurium, S. Paratyphi A, S. Kentucky, S. Inganda and S. Bargny were isolated from ducks. While, S. Virchow, S. Tamale and S. Typhimurium were isolated from Quails and S. Wingrove, finally, S. Kentucky were isolated from turkeys. Molecular characterization of common virulence genes Salmonella outer proteins (sopB), Plasmid encoded virulence gene (spvC), salmonella enterotoxin (stn) and bacterial colonization factor (bcfC) showed the presence of stn and bcfC genes in all isolates, while, sopB and Spv genes were present in 64.3% and 10.7%, respectively. It is concluded that salmonellae with common virulence genes were widely spread among domestic birds in Delta areas, Egypt, resulting in economic and public health problems which require the application of strictly biosecurity measures in poultry rearing

Prevalence of Toxigenic and Methicillin Resistant Staphylococci in Poultry Chain Production

Staphylococci are a worldwide cause of human and animal infection and are considered to be of the most common causes of infections in birds. Enterotoxins produced by some staphylococcal species were recognized as a causative agent of staphylococcal food poisoning (SFP). Only enterotoxins produced by Staphylococcus aureus were as yet well characterized. Much less is known about enterotoxigenic potential of coagulase-negative species of genus Staphylococcus (CNS). It has been reported that enterotoxigenic CNS strains have been associated with human and animal infections and food poisoning. Samples collected from chicken production cycle (un hatched eggs, baby chicks, broilers, chicken meat and table eggs) in Luxor, Egypt were tested to investigate the presence of Staphylococcus species and detection of their enterotoxines genes with more special attention for detection of methicillin resistance gene (mec A). Samples were tested for S. aureus and CNS on the basis of cultural and biochemical properties and confirmed by PCR amplification of 16S rRNA and clfa gene. Results showed that the presence of Staphylococci were 50/150 (33.3%), 14% of the samples were S. aureus (21/150), while, 19.33% were CNS (29/150). mecA gene was detected in 66.7% and 51.7% among S. aureus and CNS respectively. Enterotoxins genes (seb, sec and see) were found in 5 (23.8%) of S. aureus with a percent of (9.5%) for seb and sec and (4.8%) for see, while sec and see were found in 6 (20.6%) of CNS.  With a percent (10.3%) for each

The occurrence of disinfectant and antibiotic-resistant genes in Escherichia coli isolated from chickens in Egypt

Aim: This work aimed to determine the occurrence of antibiotic and disinfectant resistance genes in Escherichia coli isolated from chickens in Egypt. Materials and Methods: Organs (liver, lung, heart, yolk sac, and bone marrow) of 1500 chicken samples were collected from diseased chickens suffered from colibacillosis with PM findings as CRD, diarrhea and omphalitis from different governorates of Egypt as: Giza, EL-Bahira, Fayoum, El-Dakahlia, El-Ismalia, and El-Sharkia during 2015-2016. These samples were labeled and transported immediately on ice to the Reference laboratory for quality control on poultry production (RLQP). The samples were cultured onto MacConkey agar and Eosin Methylene Blue Agar. Isolation and identification of the E. coli were performed based on morphology, cultural, staining, and biochemical properties. Antimicrobial resistance test was carried out using disk diffusion method. The PCR employing tetA, qacED1 and qacA/B were carried out for detection of these genes in isolated E.coli. Results: The prevalence of E. coli in chicken was 34%. Predominant serotypes of E. coli which serologically identified were O128, O111, O44, O158, and O2. Antibiotic susceptibility test of E. coli revealed that 100% of isolates were resistant to ampicillin, erythromycin, and sulfamethoxazole-trimethoprim, while 73.53% and 38.23% of them were sensitive for colistin sulfate and levofloxacin, respectively. Antibiotic resistance genes as tetA gene were tested for isolated E. coli and detected by incidence rate of 91.18%. qac resistance genes resembling as qacED1 and qacA/B genes were detected in isolated E. coli 70.6% and 14.7%, respectively. Conclusion: E. coli isolated from chickens in Egypt was carried qac and antibiotic-resistant genes that affect the poultry industry