Prevalence, virulence factor and antimicrobial resistance analysis of Salmonella Enteritidis from poultry and egg samples in Iran.

BACKGROUND: Salmonella enterica serovar Enteritidis (S. Enteritidis) is one of the most common serovars, associated with human salmonellosis. The food-borne outbreak of this bacterium is mainly related to the consumption of contaminated poultry meat and poultry products, including eggs. Therefore, rapid and accurate detection, besides investigation of virulence characteristics and antimicrobial resistance profiles of S. Enteritidis in poultry and poultry egg samples is essential. A total of 3125 samples (2250 poultry and 875 poultry egg samples), sent to the administrative centers of veterinary microbiology laboratories in six provinces of Iran, were examined for Salmonella contamination, according to the ISO 6579 guideline. Next, duplex PCR was conducted on 250 presumptive Salmonella isolates to detect invA gene for identification of the genus Salmonella and sdf gene for identification of S. Enteritidis. Subsequently, the S. Enteritidis isolates were examined for detection of important virulence genes (pagC, cdtB, msgA, spaN, tolC, lpfC, and spvC) and determination of antibiotic resistance patterns against nalidixic acid, trimethoprim-sulfamethoxazole, cephalothin, ceftazidime, colistin sulfate, and kanamycin by the disk diffusion method. RESULTS: Overall, 8.7 and 2.3% of poultry samples and 6.3 and 1.3% of eggs were contaminated with Salmonella species and S. Enteritidis, respectively. The invA and msgA genes (100%) and cdtB gene (6.3%) had the highest and the lowest prevalence rates in S. Enteritidis isolates. The spvC gene, which is mainly located on the Salmonella virulence plasmid, was detected in 50.8% of S. Enteritidis isolates. The S. Enteritidis isolates showed the highest and the lowest resistance to nalidixic acid (87.3%) and ceftazidime (11.1%), respectively. Unfortunately, 27.0% of S. Enteritidis isolates were multidrug-resistant (MDR). CONCLUSION: The rate of contamination with Salmonella in the poultry and egg samples, besides the presence of antimicrobial resistant and MDR Salmonella isolates harboring the virulence genes in these samples, could significantly affect food safety and subsequently, human health. Therefore, continuous monitoring of animal-source foods, enhancement of poultry farm control measures, and limiting the use of antibiotics for prophylactic purposes in food producing animals, are essential for reducing the zoonotic risk of this foodborne pathogen for consumers and also choosing effective antibiotics for the treatment of salmonellosis.

The Public Health Impact of Implementing a Concentration-Based Microbiological Criterion for Controlling Salmonella in Ground Turkey.

Despite initiatives to improve the safety of poultry products in the United States, progress has stalled, and salmonellosis incidence is still above Healthy People 2020's goal. One strategy to manage Salmonella and verify process control in poultry establishments is to implement microbiological criteria (MC) linked to public health outcomes. Concentration-based MC have been used by the food industry; however, the public health impact of such approaches is only starting to be assessed. This study evaluated the public health impact of a concentration-based MC for Salmonella in raw ground turkey consumed in the United States using a quantitative risk assessment modeling approach. The distribution of Salmonella concentration in ground turkey was derived from USDA-FSIS monitoring surveys. Other variables and parameters were derived from public databases, literature, and expert opinion. Based on considered concentrations, implementing a MC of 1 cell/g led to an estimated 46.1% reduction (preventable fraction, PF) in the mean probability of illness when consumer cooking and cross-contamination were included. The PF was consistent across scenarios including or excluding cross-contamination and cooking, with slightly lower mean PF when cross-contamination was included. The proportion of lots not compliant with the 1 cell/g MC was 1.05% in the main scenarios and increased nonlinearly when higher Salmonella concentrations were assumed. Assumptions on concentration variability across lots and within lots had a large impact, highlighting the benefit of reducing this uncertainty. These approach and results can help inform the development of MC to monitor and control Salmonella in ground turkey products.

Impact of Poultry Processing Operating Parameters on Bacterial Transmission and Persistence on Chicken Carcasses and Their Shelf Life.

It is important for the poultry industry to maximize product safety and quality by understanding the connection between bacterial diversity on chicken carcasses throughout poultry processing to the end of shelf life and the impact of the local processing environment. Enumeration of total aerobic bacteria, Campylobacter and Pseudomonas, and 16S rRNA gene amplicon sequencing were used to evaluate the processing line by collecting 10 carcasses from five processing steps: prescald, postplucker, pre- and post-immersion chill, and post-air chill. The diversity throughout a 12-day shelf life was also determined by examining 30 packaged carcasses. To identify the sources of possible contamination, scald water tank, immersion chilling water tank, air samples, and wall surfaces in the air-chill room were analyzed. Despite bacterial reductions on carcasses (>5 log10 CFU/ml) throughout the process, each step altered the bacterial diversity. Campylobacter was a minor but persistent component in the bacterial community on carcasses. The combination of scalding, defeathering, and plucking distributed thermophilic spore-forming Anoxybacillus to carcasses, which remained at a high abundance on carcasses throughout subsequent processes. Pseudomonas was not isolated from carcasses after air chilling but was abundant on the wall of the air-chill room and became the predominant taxon at the end of shelf life, suggesting possible contamination through air movement. The results suggest that attention is needed at each processing step, regardless of bacterial reductions on carcasses. Changing scalding water regularly, maintaining good hygiene practices during processing, and thorough disinfection at the end of each processing day are important to minimize bacterial transmission.IMPORTANCE Culture-based and culture-independent approaches were utilized to reveal bacterial community changes on chicken carcasses at different processing steps and potential routes from the local processing environment. Current commercial processing effectively reduced bacterial loads on carcasses. Poultry processes have similar processes across facilities, but various processing arrangements and operating parameters could impact the bacterial transmission and persistence on carcasses differently. This study showed the use of a single tunnel incorporating scalding, defeathering and plucking may undesirably distribute the thermoduric bacteria, e.g., Campylobacter and Anoxybacillus, between the local environment and carcasses, whereas this does not occur when these steps are separated. The length of immersion and air chilling also impacted bacterial diversity on carcasses. Air chilling can transfer Pseudomonas from wall surfaces onto carcasses; this may subsequently influence chicken product shelf life. This study helps poultry processors understand the impact of current commercial processing and improve the chicken product quality and safety.

Rapid and sensitive detection of Salmonella with reduced graphene oxide-carbon nanotube based electrochemical aptasensor.

Rapid detection of foodborne pathogens is crucial as ingestion of contaminated food products may endanger human health. Thus, the objective of this study was to develop a biosensor using reduced graphene oxide-carbon nanotubes (rGO-CNT) nanocomposite via the hydrothermal method for accurate and rapid label-free electrochemical detection of pathogenic bacteria such as Salmonella enterica. The rGO-CNT nanocomposite was characterized using Fourier transform infrared spectroscopy, Raman spectroscopy, X-ray diffraction and transmission electron microscopy. The nanocomposite was dropped cast on the glassy carbon electrode and further modified with amino-modified DNA aptamer. The resultant ssDNA/rGO-CNT/GCE aptasensor was then used to detect bacteria by using differential pulse voltammetry (DPV) technique. Synergistic effects of aptasensor was evident through the combination of enhanced electrical properties and facile chemical functionality of both rGO and CNT for the stable interface. Under optimal experimental conditions, the aptasensor could detect S. Typhimurium in a wide linear dynamic range from 101 until 108 cfu mL-1 with a 101 cfu mL-1 of the limit of detection. This aptasensor also showed good sensitivity, selectivity and specificity for the detection of microorganisms. Furthermore, we have successfully applied the aptasensor for S. Typhimurium detection in real food samples.

Changes of the bacterial community diversity on chicken carcasses through an Australian poultry processing line.

Understanding the bacterial community profile through poultry processing could help the industry to produce better poultry products. In this study, 10 chicken carcasses were randomly sampled from before and after scalding, before and after immersion chilling, and after air chilling each through a modern commercial processing line, along with the contents of 10 caeca. The sampled processing line effectively reduced the bacterial counts by > 4.6 Log10 CFU/ml for each of Total Viable Counts, Escherichia coli and Campylobacter. However, the metagenomics results suggested that Lactobacillus, Staphylococcus and unclassified Lachnospiraceae persisted at all sampling stages. Pseudomonas, Paeniglutamicibacter, Chryseobacterium and Pseudarthrobacter comprised 47.2% in the bacterial community on samples after air chilling compared to 0.3% on samples after immersion chilling, whereas TVCs were the same. Overall, the current interventions of the investigated poultry processing line were unable to eliminate persistence of certain foodborne pathogens, despite a significant reduction of the overall bacterial counts. Chilling is an important controlling point in contamination/cross-contamination, particularly extended air chilling. Lastly, the large presence of Pseudomonas on chickens after air chilling may lead to downstream spoilage related issues, which needs more investigation to explore quantitatively the effect on the shelf life of poultry products.

Efficient isolation of Campylobacter bacteriophages from chicken skin, analysis of several isolation protocols.

The application of Campylobacter specific bacteriophages appears as a promising food safety tool for the biocontrol of this pathogen in the poultry meat production chain. However, their isolation is a complicated challenge since their occurrence appears to be low. This work assessed the efficiency of seven protocols for recovering Campylobacter phages from chicken skin samples inoculated at phage loads from 5.0 × 101 to 5.0 × 106 PFU/g. The enrichment of chicken skin in selective Bolton broth containing target isolates was the most efficient procedure, showing a low detection limit of 5.0 × 101 PFU/g and high recovery rates of up to 560%. This method's effectiveness increased as phage concentration decreased, showing its suitability for phage isolation. When this method was applied to isolate new Campylobacter phages from retail chicken skin, a total of 280 phages were recovered achieving an isolation success rate of 257%. From the 109 samples 68 resulted phage positive (62%). Chicken skin could be, therefore, considered a rich source in Campylobacter phages. This method is a simple, reproducible and efficient approach for the successful isolation of both group II and III Campylobacter specific bacteriophages, which could be helpful for the enhancement of food safety by reducing this pathogen contamination in broiler meat.

Microfluidic based impedance biosensor for pathogens detection in food products.

A MEMS-based impedance biosensor was designed, fabricated, and tested to effectively detect the presence of bacterial cells including E. coli O157:H7 and Salmonella typhimurium in raw chicken products using detection region made of multiple interdigitated electrode arrays. A positive dielectrophoresis based focusing electrode was used in order to focus and concentrate the bacterial cells at the centerline of the fluidic microchannel and direct them toward the detection microchannel. The biosensor was fabricated using surface micromachining technology on a glass substrate. The results demonstrate that the device can detect Salmonella with concentrations as low as 10 cells/mL in less than 1 h. The device sensitivity was improved by the addition of the focusing electrodes, which increased the signal response by a factor between 6 and 18 times higher than without the use of the focusing electrodes. The biosensor is selective and can detect other types of pathogen by changing the type of the antibody immobilized on the detection electrodes. The device was able to differentiate live from dead bacteria.

Phentotypic, gentotypic antimicrobial resistance and pathogenicity of Salmonella enterica serovars Typimurium and Enteriditis in poultry and poultry products.

For detection and isolation of Salmonella enterica, 650 meat and tissue samples were processed using Rappaport-Vassiliadis Enrichment broth and Salmonella Chromogenic agar followed by confirmation through specific antisera and polymerase chain reaction (PCR) targeting their Specific Serovar Genomic Regions (SSGRS). Isolates were tested for 15 antibiotics (CRO, AMX, GEN, STR, TET, CHL, CLR, LVX, OFX, GAT, CIP, SXT, AMP, LIN and AZM) according to the disc diffusion method and antimicrobial resistant genes (tet(A), tet(B), tet(C), strA/strB, aadA, aac(3)IV), aadB, sul1, sul2 and sul3, blaCMY-2, blaTEM and blaSHV) using PCR. The overall prevalence of Salmonella enterica was 12%, being higher in markets (15%) as compared to poultry farms (37.2%). The MPN of all positive meat and tissue samples was found 3.6 MPN/gram (0.17-18). A total of 234 isolates were obtained, serovar Typimurium (139) and Enteridits (95) were the most prevalent. Antimicrobial resistance patterns were different in different serovars according to origin of Salmonella isolates. The overall isolates were highly resistant for LIN (93.1%, 218/234) followed by AMX (80%, 187/234), AMP (74.3%, 174/234), TET (64.5%, 151/234) and STR (64.5%, 151/234). Overall, the most common ARG was blaTEM (76%, 178/234), followed by blaSHV (71.7%, 168/234), tet(A) (64%, 151/234) and tet(B) (64%, 150/234), while the least ARG was aadB (7.2%, 17/234). Both Typimurium and Enteridits were tested in the Balb/C mice for pathogenicity. Both Typimurium and Enteridits were found to cause successful colonization, 100% morbidity but Enteriditis were found to cause 33% mortality.

Overexpressing ovotransferrin and avian ß-defensin-3 improves antimicrobial capacity of chickens and poultry products.

Zoonotic and foodborne diseases pose a significant burden, decreasing both human and animal health. Modifying chickens to overexpress antimicrobials has the potential to decrease bacterial growth on poultry products and boost chicken innate immunity. Chickens overexpressing either ovotransferrin or avian ß-defensin-3 (AvßD3) were generated using Tol-2 transposons. Transgene expression at the RNA and protein level was seen in egg white, breast muscle, and serum. There were significant differences in the immune cell populations in the blood, bursa, and spleen associated with transgene expression including an increased proportion of CD8+ cells in the blood of ovotransferrin and AvßD3 transgenic birds. Expression of the antimicrobials inhibited the in vitro growth of human and chicken bacterial pathogens and spoilage bacteria. For example, transgene expression significantly reduced growth of aerobic and coliform bacteria in breast muscle and decreased the growth of Salmonella enterica in egg white. Overall these results indicate that overexpression of antimicrobials in the chicken can impact the immune system and increase the antimicrobial capacity of poultry products.

Inactivation of extraintestinal pathogenic E. coli clinical and food isolates suspended in ground chicken meat by gamma radiation.

Extraintestinal pathogenic Escherichia coli are common contaminants in retail poultry and involved inflammatory bowel disease, urinary tract infections and meningitis in both animals and humans. They cause significantly more illnesses and deaths in humans than Shiga toxin-producing E. coli (STEC). Ionizing radiation is used commercially for improving the safety and shelf-life of foods. In this study we inoculated ground chicken meat with 25 individual isolates of clinical uropathogenic E. coli (UPEC) and newborn meningitis causing E. coli (NMEC), isolates from retail chicken meat (CM), as well as retail chicken-skin isolates identified in our laboratory (CS). We then determined their gamma radiation inactivation kinetics (D10-value). The mean D10-value for all isolates (n = 25) was 0.30 kGy. The mean D10-value for the UPEC, NMEC, CM, and CS isolates were 0.25, 0.29, 0.29, and 0.39 kGy, respectively. The mean D10-value for the clinical isolates was 0.27 kGy vs. 0.34 kGy for the non-clinical isolates. There was no correlation between presence of virulence factors, antibiotic resistance, and radiation resistance. ExPEC were similar to that of STEC which were previously evaluated in our laboratory. The radiation doses needed to kill STEC poultry meat should also kill ExPEC.

Contamination of yellow-feathered broiler carcasses: Microbial diversity and succession during processing.

The processing environment of broiler processing plants is a potential major source of bacterial contamination of broiler carcasses. This study investigated the effect of processing water and processing time on the microbial diversity of yellow-feathered broiler carcasses at select stages of slaughter during one commercial processing day using a high-throughput sequencing technique targeting the V3V4 region of the 16S RNA gene. Our results demonstrated that Firmicutes and Proteobacteria were the dominant bacterial phyla of broiler carcasses and processing water in the chiller tank, whereas the processing water in the scalder tank contained a high abundance of Firmicutes and Deinococcus-Thermus. At the genus level, Escherichia-Shigella and Streptococcus were present on broiler carcasses with high abundances after defeathering, but their abundance decreased after washing and chilling. The bacterial community structure was revealed to become more complex at later stages of processing, as indicated by the consistent increase in microbial alpha diversity metrics (Chao 1, Shannoneven and Shannon) throughout the processing stages (p < 0.05). Significantly separate clustering of bacterial communities between scalder tank water and carcasses was revealed by PCoA analysis, indicating the limited effect of scalding water on the bacterial communities of broiler carcasses.

Transcriptional profiling and metabolomic analysis of Staphylococcus aureus grown on autoclaved chicken breast.

Although Staphylococcus aureus is a major cause of food poisoning, little is known about its response to growth on food. Utilizing a transcriptional profiling and metabolomics approach, we compared S. aureus grown on autoclaved chicken breast (ACB) to Luria broth agar. ACB cultures demonstrated increased expression of genes associated with protein synthesis, cofactors, secondary metabolites, nitrogen and nucleotide metabolism, amino acid transport, and reduced expression of general stress, lipid metabolism, and virulence genes. The ACB culture also displayed characteristics of catabolite de-repression and anaerobic growth, and increased expression of arginine biosynthesis genes (argFGH) and an arginine/ornithine antiporter gene (arcD). S. aureus synthesizes arginine from proline and the ACB culture exhibited increased expression of proline transport genes (opuBA, opuBB and putP) and increased proline accumulation. Amino acid and sugar content in the ACB grown culture increased, and this was attributed to the consumption of ACB, transport of amino acids, and gluconeogenesis. Genes involved with biotin biosynthesis and uptake were upregulated and biotin is required for amino acid catabolism. Genes encoding urease and urease activity were upregulated in ACB cultures, while urea levels were reduced. This research provides fundamental information on the response of S. aureus growing on chicken meat that could find application in future attempts to reduce the growth of S. aureus in food.

Antimicrobial Resistance Profile of Salmonella Isolates in Chicken Carcasses in Dakar, Senegal.

The incidence of the Salmonella contamination of poultry products in Senegal is unknown. Salmonella contamination and antimicrobial drug resistance profiles in chicken carcasses were investigated. Between July 2012 and July 2013, three types of chicken carcasses (broilers, laying hens, and premises chickens) obtained from retailers in the markets of Dakar and its suburbs were tested for Salmonella contamination. Salmonella strains were isolated from 300 chicken carcasses according to International Organization for Standardization ISO 6579 (2002) guidelines. In these samples, 273 isolates were obtained, belonging to 22 serovars, and 53% samples were contaminated with at least 1 serovar. Standardized techniques were used for the susceptibility testing and serotyping of isolates. Hygiene conditions, in terms of the cleanliness of stalls, the packing of chicken carcasses in bags, and the maintenance of the cold chain at the stall, were moderately poor. The three serovars most frequently identified were Salmonella Istanbul (28%), Salmonella Brancaster (19%), and Salmonella Kentucky (13%). Overall, 21% of isolates were resistant to quinolones and fluoroquinolones. Serovar Istanbul was resistant to tetracycline (TE) and trimethoprim + sulfamethoxazole (SXT). Serovars Brancaster and Kentucky were resistant to betalactams and to quinolones or fluoroquinolones. The uncommon serovar Senftenberg had the strongest resistance profile, displaying resistance to betalactams including imipenem (IMP). Large numbers of isolates were resistant to TE (66%) and SXT (47%). Resistance to cephalosporins (5%), chloramphenicol (2%), gentamicin (8%), and IMP (1%) was less frequent. A large proportion of the broilers sold in Dakar markets were contaminated with Salmonella. This situation probably resulted from poor hygiene conditions in chicken farms and slaughterhouses and from breaks in the cold chain at some point in the distribution of poultry products.

Alpha-cyclodextrin as a new functional ingredient in low-fat chicken frankfurter.

1. The current consumer preference for healthier meat products is associated with less additives in manufacturing (so-called 'clean-label') or the addition of non-meat ingredients with functional properties, recognised as improving specific technological properties in meat products.2. This study evaluated the effect of the addition of alpha-cyclodextrin and wheat fibre to low-fat chicken frankfurters containing 35% mechanically deboned chicken meat on the technological and sensorial properties during refrigerated storage.3. The results showed that the addition of dietary fibres (alpha-cyclodextrin and wheat fibre) in low-fat chicken frankfurters improved emulsion stability, hardness, chewiness and reduced cohesiveness.4. Alpha-cyclodextrin helped the retention of fat globules in the microstructure and affected colour in the sensorial evaluation.5. The use of alpha-cyclodextrin, in combination with wheat fibre, as a new ingredient to substitute fat in emulsified meat products containing mechanically deboned chicken, improved emulsion stability and texture.6. Alpha-cyclodextrin and wheat fibre were effective in contributing to fat reduction without affecting the sensory properties of the product.

Growth and Persistence of Campylobacter jejuni in Foodstuffs.

Campylobacter genus bacteria causing campylobacteriasis are difficult to culture. This fact necessitates creation of special approaches to studies of the behavior of these pathogens during the manufacture and storage of foodstuffs. The regularities of Campylobacter jejuni transition into an uncultivable state are studied under conditions simulating the process of immersion cooling of fresh poultry products. The proportion of viable colony-forming (CFU) cells to the total count of planktonic and uncultivable cells in the population was calculated by the level of genomic DNA in the samples evaluated by quantitative real-time PCR with intercalating dyes. PCR was carried out with primers detecting the cytolethal toxin subunit B gene cdtB and invasion gene ciaB in C. jejuni strains. The count of detected cells was 5-10-fold higher than the count of CFU; the cultural method failed to detect the agent in 40% analyzed samples of superficially infected products, while the level of uncultivable cells detected by PCR was significantly higher. The relationship between culturing conditions and formation of C. jejuni biofilms was studied. The most intensive formation of film exomatrix was observed under unfavorable conditions for this microorganism at 25oC. In microaerophilic gaseous medium, weak formation of films and intensive growth of C. jejuni populations were observed. Culturing at higher temperatures (37-42oC) was in fact inessential for the film formation process.

Whole-Genome Sequencing and Bioinformatic Analysis of Isolates from Foodborne Illness Outbreaks of Campylobacter jejuni and Salmonella enterica.

Whole-genome sequencing (WGS) via next-generation sequencing (NGS) technologies is a powerful tool for determining the relatedness of bacterial isolates in foodborne illness detection and outbreak investigations. WGS has been applied to national outbreaks (for example, Listeria monocytogenes); however, WGS has rarely been used in smaller local outbreaks. The current study demonstrates the superior resolution of genetic and evolutionary relatedness generated by WGS data analysis, compared to pulsed-field gel electrophoresis (PFGE). The current study retrospectively applies WGS and a reference-free bioinformatic analysis to a Utah-specific outbreak of Campylobacter jejuni associated with raw milk and to a national multistate outbreak of Salmonella enterica subsp. enterica serovar Typhimurium associated with rotisserie chicken, both of which were characterized previously by PFGE. Together, these analyses demonstrate how a reference-free WGS workflow is not reliant on determination of a reference sequence, like WGS workflows that are based on single-nucleotide polymorphisms, or the need for curated allele databases, like multilocus sequence typing workflows.

Extended-spectrum cephalosporin-resistant Escherichia coli isolated from chickens and chicken meat in Brazil is associated with rare and complex resistance plasmids and pandemic ST lineages.

Objectives: Brazil is the greatest exporter of chicken meat (CM) in the world. It is of utmost importance to monitor resistance to extended-spectrum cephalosporins (ESCs) in this sector because resistance to ESCs in Escherichia coli isolated from food-producing animals may contaminate humans through the food chain. Thus, the aim of this study was to characterize and compare ESC-resistant E. coli isolated from chickens and retail CM produced in south-eastern Brazil. Methods: Five CM samples and 117 chicken cloacal swabs (CCSs) were inoculated on MacConkey agar supplemented with cefotaxime. Presumptive E. coli colonies were identified and antimicrobial susceptibility was tested. Virulence and acquired blaESBL and blaAmpC genes were sought and genetic environments characterized. Isolates were typed by phylogenetic grouping, XbaI-PFGE and MLST. Results: All five CM samples and 36 CCSs (30.8%) were positive for the presence of ESC-resistant E. coli, leading to the selection of 58 resistant isolates. ESC resistance was mostly due to the presence of the chromosome-encoded blaCTX-M-2 gene, but plasmid-mediated blaCTX-M-2, blaCTX-M-8, blaCTX-M-15, blaCTX-M-55 and blaCMY-2 were also detected. Multireplicon plasmids were sporadically identified, such as IncHI2/P-blaCTX-M-2 and IncFII/N-blaCTX-M-55. Phylogroup D predominated, while PFGE and MLST revealed a high genetic diversity. Conclusions: Live Brazilian chickens and CM act as reservoirs of ESC-resistant E. coli and resistance genes are located on highly diverse genetic determinants. Potentially pathogenic strains, which may represent a threat to human health and a source of environmental contamination, were also identified. Active surveillance is therefore essential in Brazil's chicken production line.

Virulence genes and genetic diversity assessment of Shiga toxin-producing Escherichia coli O91 strains from cattle, beef and poultry products.

Shiga toxin-producing Escherichia coli (STEC) O91 has ranked in the top five of the non-O157 serogroups most frequently associated with human cases. In order to gain insight into the genetic diversity of O91 Latin American STEC strains, we analyzed their virulence properties and carried out a subtyping assay. A panel of 21 virulence genetic markers associated with human and animal infections was evaluated and the relatedness among strains was determined by a multiple-locus variable-number tandem repeats analysis (MLVA) comprising 9 VNTR loci. Twenty-two STEC O91 isolated from cattle and meat food and belonging to 5 serotypes (O91:H21, O91:H8, O91:H14, O91:H28, O91:H40) were studied. Eight virulence profiles were obtained for the O91 STEC strains: 4 for O91:H21 plus one for O91:H8, O91:H14, O91:H28 and O91:H40. All strains contained ehxA and lpfA0113 genes and only both stx1-positive strains lacked saa, which encodes the STEC autoagglutinating adhesin. Other genes involved in adhesion were detected: ehaA (91%), elfA and espP (86%), ecpA (82%) and, hcpA (77%). The gene encoding the cytolethal distending toxin type-V (CDT-V) was found only in O91:H8 and O91:H21, being present in the majority (89%) of strains of this last serotype. MLVA typing divided the total number of strains into 12 genotypes, and 9 of them were unique to a single strain. No association was observed between the virulence profiles and the source of the strains. Although they lack the eae gene, most of the strains have the genetic potential to adhere to host cells through other structures and possess cdt-V, which has been found in STEC strains involved in serious diseases. The MLVA showed clonal relatedness among strains isolated from cattle belonged to a same dairy farm and suggested that the same clone remains circulating throughout the year and, on the other hand, the need to increase the number of VNTR loci which could allow a higher discrimination among O91:H21 isolates.

Trends in microbial control techniques for poultry products.

Fresh poultry meat and poultry products are highly perishable foods and high potential sources of human infection due to the presence of several foodborne pathogens. Focusing on the microbial control of poultry products, the food industry generally implements numerous preventive measures based on the Hazard Analysis and Critical Control Points (HACCP) food safety management system certification together with technological steps, such as refrigeration coupled to modified atmosphere packaging that are able to control identified potential microbial hazards during food processing. However, in recent years, to meet the demand of consumers for minimally processed, high-quality, and additive-free foods, technologies are emerging associated with nonthermal microbial inactivation, such as high hydrostatic pressure, irradiation, and natural alternatives, such as biopreservation or the incorporation of natural preservatives in packaging materials. These technologies are discussed throughout this article, emphasizing their pros and cons regarding the control of poultry microbiota and their effects on poultry sensory properties. The discussion for each of the preservation techniques mentioned will be provided with as much detail as the data and studies provided in the literature for poultry meat and products allow. These new approaches, on their own, have proved to be effective against a wide range of microorganisms in poultry meat. However, since some of these emergent technologies still do not have full consumer's acceptability and, taking into consideration the hurdle technology concept for poultry processing, it is suggested that they will be used as combined treatments or, more frequently, in combination with modified atmosphere packaging.

Campylobacter control measures in indoor broiler chicken: critical re-assessment of cost-utility and putative barriers to implementation.

As campylobacteriosis is one of the most important foodborne infections, a European Union (EU)-27 level cost-effectiveness model has been developed on the socio-economic costs and benefits of applying certain control measures for the reduction of Campylobacter in broiler meat. This is expected to be a gold standard for food safety policymakers in the EU; hence, the validity of its modelling assumptions is essential. The authors of the present paper conducted an independent review of model input parameters on health and economic burden and found that the model most probably overestimated the burden of human campylobacteriosis. A discounted, quality-adjusted life year (QALY)-based European estimate has been developed for human campylobacteriosis and resulted in 15.23 QALY loss per 1000 human gastroenteritis cases. Country-specific cost of illness estimates have been developed for various countries in the EU-27. Based on these model adaptations, a selected Campylobacter control strategy was re-assessed and its high cost-effectiveness was confirmed at the EU level, and also in all but three Member States. Bacteriocin treatment or vaccination of the animals, two alternative control measures were also re-evaluated, and these strategies seemed to be far less cost-effective than the investigated strategy. Putative barriers to the rapid implementation of the investigated Campylobacter control strategy are discussed, and potential solutions are proposed. Further research is required on stakeholder perspectives pertaining to the realistic barriers and implementation opportunities.