Isolation, characterization, and resistance profile of Salmonella spp. from chicken cuts / Isolamento, caracterização e perfil de resistência de Salmonella spp. provenientes de cortes de frango

The present study aimed at isolating and characterizing Salmonella spp. from chicken cuts marketed in Francisco Beltrão, PR, and verify the resistance profile of the isolates against antimicrobials used in human therapy. Samples of chicken cuts (n=40) were purchased from supermarkets and submitted to microbiological analysis for the detection of Salmonella spp. The suspected colonies underwent biochemical testing for the identification of enterobacteria. Four colonies were selected from each sample positive for Salmonella spp., totaling 28 isolates that were tested for antimicrobial sensitivity. Colonies that showed resistance to ceftriaxone were subjected to extended-spectrum beta-lactamases (ESBL). Among the analyzed chicken samples, seven (17.5%) showed biochemical behavior characteristic of Salmonella spp. Among the 28 isolates, seventeen different resistance profiles were found, of which 46.42% (n=13) had a multi-resistance profile, and 21.4% (n=6) of the isolates had a phenotype for ESBL production. The strains of Salmonella spp. isolated from chicken cuts found in this study showed a high level of resistance to antimicrobials of different classes and of last generations, these data serve as a warning, as they put the human treatment of salmonellosis at risk.(AU)

A pesquisa objetivou isolar e caracterizar Salmonella spp., a partir de cortes de frangos comercializados na cidade de Francisco Beltrão - PR, bem como verificar o perfil de resistência dos isolados em relação aos antimicrobianos utilizados na terapêutica humana. Amostras de cortes de frango (n=40) foram adquiridas em supermercados e submetidas à análise microbiológica para detecção de Salmonella spp. As colônias suspeitas foram submetidas a provas bioquímicas para identificação de enterobactérias. Quatro colônias foram selecionadas de cada amostra positiva para Salmonella spp., totalizando 28 isolados, que foram testadas quanto à sensibilidade a antimicrobianos. As colônias que apresentaram resistência à ceftriaxona foram submetidas à pesquisa de betalactamases de espectro estendido (ESBL). Das amostras de frango analisadas, sete (17,5%) apresentaram comportamento bioquímico característico de Salmonella spp. Entre os 28 isolados, foram encontrados 17 perfis diferentes de resistência, tendo 46,42% (n=13) apresentado perfil de multirresistência e 21,4% (n=6) apresentado fenótipo para produção de ESBL. As cepas de Salmonella spp. isoladas de cortes de frango, encontradas neste estudo, apresentaram alto índice de resistência a antimicrobianos de diferentes classes e de últimas gerações. Esses dados servem de alerta, uma vez que coloca em risco o tratamento da salmonelose humana.(AU)

Detection of Campylobacter jejuni and Salmonella typhimurium in chicken using PCR for virulence factor hipO and invA genes (Saudi Arabia).

Campylobacter jejuni and Salmonella typhimurium are the leading causes of bacterial food contamination in chicken carcasses. Contamination is particularly associated with the slaughtering process. The present study isolated C. jejuni and S. typhimurim from fifty chicken carcass samples, all of which were acquired from different companies in Riyadh, Saudi Arabia. The identification of C. jejuni was performed phenotypically by using a hippurate test and genetically using a polymerase chain reaction with primers for 16S rRNA and hippurate hydrolase (hipO gene). For the dentification of S. typhimurim, a serological Widal test was carried out using serum anti-S. typhimurium antibodies. Strains were genetically detected using invA gene primers. The positive isolates for C. jejuni showed a specific molecular size of 1448 bp for 16S rRNA and 1148 bp for hipO genes. However, the positive isolates of the invA gene exhibited a specific molecular size at 244 bp using polymerase chain reaction (PCR). Comparing sequencing was performed with respect to the invA gene and the BLAST nucleotide isolates that were identified as Salmonella enterica subsp. enterica serovar typhimurium strain ST45, thereby producing a similarity of 100%. The testing identified C.jejuni for hippuricase, GenBank: Z36940.1. While many isolates of Salmonella spp. that contained the invA gene were not necessarily identified as S. typhimurim, the limiting factor for the Widal test used antiS. typhimurum antibodies. The multidrug resistance (MDR) of C. jejuni isolates in chickens was compared with the standard C. jejuni strain ATCC 22931. Similarly, S. typhimurium isolates were compared with the standard S. typhimurium strain ATCC 14028.

Characterization of Pediococcus acidilactici postbiotic and impact of postbiotic-fortified chitosan coating on the microbial and chemical quality of chicken breast fillets.

This study was carried out to characterize antioxidant activity, total phenolic content, and the phenolic and flavonoids profile of postbiotic of Pediococcus acidilactici and to evaluate the effects of postbiotics (10% and 50%) alone and in combination with chitosan coating (1%) on the microbial and chemical quality of chicken breast fillets during storage at 4 °C. Antioxidant activity and total phenolic content of the postbiotics were found to be 1291.02 ± 1.5 mg/L TEAC and 2336.11 ± 2.36 mg/L GAE, respectively. The most abundant phenolic was vanillic acid, followed by t-caffeic, gallic, and caftaric acids. The postbiotic-chitosan (50% + 1%) combination decreased L. monocytogenes and S. Typhimurium counts by 1.5 and 2.1 log10 CFU/g, respectively, compared to the control (P < 0.05). This combination decreased the total viable count (TVC), lactic acid bacteria (LAB), and psychrotrophic bacteria count compared to the control (P < 0.05). No differences were found in thiobarbituric acid (TBA) values among the samples during storage (P > 0.05). Postbiotic treatment did not significantly change the pH values and color properties of the breast fillets (P > 0.05). Postbiotic-chitosan combinations extended the shelf-life by up to 12 days compared to the control. In conclusion, the postbiotic-chitosan combination can be used to preserve and improve the microbial quality of chicken meat products.

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.

Selection and application of natural antimicrobials to control Clostridium perfringens in sous-vide chicken breasts inhibition of C. perfringens in sous-vide chicken.

Current consumer preferences for both clean label food ingredients and convenience-based foods has provided a unique opportunity to explore the application of novel natural food preservatives in sous vide products. The anaerobic environment and relatively low thermal processing of the sous vide process creates a favorable environment for the survival, germination, and outgrowth of spore-forming bacterium Clostridium perfringens. The aim of this study was to identify effective novel natural ingredient formulations against C. perfringens and apply them within a vacuum-sealed sous vide chicken model exposed to abusive storage and chilling conditions. Among six commercial vinegar-based formulations, liquid vinegar with citrus extract (CE; 1.0%) and with lemon juice concentrate (LJC; 1.5%) were identified as the most effective at inhibiting three individual C. perfringens strains. Both reduced viable cell counts by 5 log CFU/mL (P < 0.05), whereas reductions in spore counts ranged from 2 to 4 log CFU/mL depending on formulation and concentration used. Once incorporated to chicken meat 1.0% CE and 1.5% LJC before sous-vide cooking, completely inhibited the growth of mixed C. perfringens strains (P < 0.05) during storage for 16 days at 12 and 16 °C. Exponential cooling from 54 to 4 °C was performed for 18 h to imitate abusive storage conditions. CE and LJC at 3.0% inhibited growth and reduced counts by 3.4 and 2.9 log CFU/g compared to respective controls. Treatments CE and LJC could be implemented within the formulation of a sous vide chicken product to provide an effective protection against C. perfringens meeting clean label expectations.

Nanocomposite active packaging based on chitosan biopolymer loaded with nano-liposomal essential oil: Its characterizations and effects on microbial, and chemical properties of refrigerated chicken breast fillet.

Biodegradable films reinforced with bio-nanomaterials are a solution for developing active packaging systems, shelf-life extension and protection of environment against conventional packaging. This study aimed to characterize the biocompatible chitosan (CS) films formulated with nano-liposomal garlic essential oil (NLGEO) and assess the physicho-mechanical, morphology properties and also microbial and chemical changes in chicken fillets during storage time at 4 °C. NLGEO was obtained by thin-layer hydration-sonication method using glycerol and tween 80 as plasticizer and emulsifier, respectively. Different levels (0, 0.5, 1 and 2%) of NLGEO with average size of ~101 nm were added into the chitosan matrix and films fabricated by casting method. The average size, polydispersity index and zeta potential were ~101 nm, 0.127 and -7.23, respectively. Control samples showed higher values for pH, total volatile nitrogen (TVN), peroxide value (PV), thiobarbituric acid-reactive substances (TBARS), and microbial count including total viable count (TVC), coliforms, Staphylococcus aureus and psychrotroph bacteria than treated samples. The films with higher NLGEO content represented stronger inhibitory effects. The incorporation of NLGEO improved the mechanical properties and water resistance of active films. Microstructure analysis also showed a nearly smooth surface morphology and homogenous structure with a good dispersion for NLGEO films. Significant synergistic effects in chemical and bacterial preservation of chicken fillet samples were observed by NLGEO films. The optimal mechanical and barrier properties of chitosan-NLGEO films introduced it a potential active packaging to extend the shelf life of chicken fillet.

Comparative history of Campylobacter contamination on chicken meat and campylobacteriosis cases in the United States: 1994-2018.

In many countries campylobacteriosis ranks as one of the most frequently reported foodborne illnesses and poultry is the commodity that is most often associated with these illnesses. Nevertheless, efforts to reduce the occurrence of pathogen contamination on poultry are often more focused on Salmonella. While some control measures are pathogen specific, such as pre-harvest vaccination for Salmonella, improvements in sanitary dressing and interventions applied during the slaughter process can be effective against all forms of microbial contamination. To investigate the potential effectiveness of these non-specific pathogen reduction strategies in the United States, it is helpful to assess if, and by how much, Campylobacter contamination of chicken meat has changed across time. This study assesses change considering data collected in both slaughter and retail establishments and comparing observed trends in contamination with trends in human surveillance data. The results support the assertion that substantial reductions in Campylobacter contamination of chicken meat in the late 1990s and early 2000s contributed to a reduction in the human case rate of campylobacteriosis. Further reductions in chicken meat contamination between 2013 and 2018 are more difficult to associate with trends in human illnesses, with one contributing factor being the inclusion of culture independent diagnostic test results in the official case counts during that time. Other contributing factors are discussed.

Strategies to Improve Poultry Food Safety, a Landscape Review.

Food safety remains a significant public health issue for the poultry industry. Foodborne pathogens can be in contact at all phases of poultry production, from initial hatch to processing and ultimately to retail and meal preparation. Salmonella and Campylobacter have been considered the primary foodborne pathogens associated with poultry. Both organisms are major causative agents of human foodborne illness. Limiting these pathogens in poultry production requires identifying their sources and routes of transmission. This involves the ability to isolate and precisely identify them using methodologies capable of discernment at the genome level. Interventions to reduce their occurrence in poultry production employ two basic strategies: prevention of establishment and elimination of already-established pathogens. This review provides an overview of current findings and prospects for further research on poultry food safety issues.

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.

Indigenous Bacteriocin of Lactic Acid Bacteria from "Dadih" a Fermented Buffalo Milk from West Sumatra, Indonesia as Chicken Meat Preservative.

BACKGROUND AND OBJECTIVE: The bacteriocin isolated from fermented buffalo milk from West Sumatra-Indonesia, called Dadih, can be considered as a natural and safe antimicrobial compound for food products. The objective of this research was to evaluate the antimicrobial activity of bacteriocin from Dadih and its effectiveness as a preservative in chicken meat. MATERIALS AND METHODS: This study used experimental method followed by statistical analysis using 3 experiments with duplication including experiment of meat samples (0 and 10% bacteriocin), storage temperatures (7 and 26°C) and storage duration (0, 1, 2, 3, 4, 5, 6 days and 0, 6, 12 hrs). Each experiment consists of a bacteriocin test, antimicrobial activity assay, physicochemical measurement and storability. RESULTS: From 10 LAB isolates successfully obtained from Dadih, two isolates with D7 code and D10 code had the highest antimicrobial activity, reaching 11.75 mm and 12 mm, respectively. The meat treated by 10% of bacteriocin gave the lower total microbial (3rd and 5th day) and total E. coli (5th day) at 7 and 26°C. The pH and water activity (aw) values of chicken meat with 10% of bacteriocin showed lower values at 7 and 26°C. The application of bacteriocin to chicken meat was able to inhibit the microbial growth that was still below standard for 3 days at 7°C and 6 hrs at 26°C. CONCLUSION: Based on research, lactic acid bacteria isolated from buffalo milk curd produced bacteriocin compound which has antimicrobial properties. This bacteriocin showed potential as a natural preservative for chicken meat by inhibiting the growth of pathogen microorganisms.

Accurate and sensitive detection of Salmonella in foods by engineered bacteriophages.

Salmonella is a major causative agent of foodborne illness and rapid identification of this pathogen is essential to prevent disease. Currently most assays require high bacterial burdens or prolonged enrichment to achieve acceptable performance. A reduction in testing time without loss of sensitivity is critical to allow food processors to safely decrease product holding time. To meet this need, a method was developed to detect Salmonella using luciferase reporter bacteriophages. Bacteriophages were engineered to express NanoLuc, a novel optimized luciferase originating from the deep-sea shrimp Oplophorus gracilirostris. NanoLuc-expressing bacteriophages had a limit of detection of 10-100 CFU per mL in culture without enrichment. Luciferase reporters demonstrated a broad host range covering all Salmonella species with one reporter detecting 99.3% of 269 inclusivity strains. Cross-reactivity was limited and only observed with other members of the Enterobacteriaceae family. In food matrix studies, a cocktail of engineered bacteriophages accurately detected 1 CFU in either 25 g of ground turkey with a 7 h enrichment or 100 g of powdered infant formula with a 16 h enrichment. Use of the NanoLuc reporter assay described herein resulted in a considerable reduction in enrichment time without a loss of sensitivity.

Stress resistance of emerging poultry-associated Salmonella serovars.

In recent years, the on-farm prevalence of some poultry-related Salmonella serovars such as S. Kentucky, S. Heidelberg, S. Livingstone and S. Mbandaka has increased significantly, even replacing S. Enteritidis and S. Typhimurium as the most frequently isolated serovars in some production settings and countries. For this reason, the aim of this work was to determine the resistance to several stressing agents and food preservation technologies, in laboratory media and in egg products, of 4 strains of these emerging Salmonella serovars associated to poultry and poultry products and to make comparisons with 4 S. Enteritidis strains. First, the resistance to acid pH, hydrogen peroxide, NaCl, heat, HHP, PEF and UV of the 8 Salmonella strains studied was determined and compared in laboratory media. From this part of the study, it was concluded that variability in resistance to stress among the 8 studied strains varied depending on the investigated agent/technology. However, differences in resistance (2D-values) were always lower than 3.3-fold. Results obtained also indicated that the strains of the emerging serovars studied would display lower acid and NaCl resistance, higher heat resistance and similar oxidative, HHP, PEF and UV resistance than S. Enteritidis. Then, the resistance of these 8 strains was evaluated and compared in egg, egg products and poultry manure. For some agents -including osmotic stresses, UV and PEF- there was a very good correspondence between the results obtained in laboratory media and in real food matrices and poultry manure (r > 0.85; p < 0.01). A significant relationship was also found for acid and HHP resistance (p < 0.05) and a trend for heat resistance (p < 0.10). Therefore, in general terms, conclusions drawn from the study carried out in laboratory media - regarding intraspecific variability and the relative resistance of the different strains - might be extrapolated, although with caution, to real food scenarios. Results obtained in this investigation would help to better understand the physiology and ecology of Salmonella and to design better egg preservation strategies.

Prevalence and antimicrobial susceptibility of Salmonella in poultry farms and in-contact humans in Adama and Modjo towns, Ethiopia.

Consumption of contaminated poultry and poultry products represents a common source of nontyphoidal Salmonella infection. Little is known on the status of Salmonella and their antimicrobial susceptibility in poultry farms in Ethiopia. This study investigated the prevalence, serotype distribution, and antimicrobial susceptibility of nontyphoidal Salmonella among poultry farms in Adama and Modjo towns. Three hundred thirty-four cloacal swabs, 384 fecal droppings of birds, 59 feed, 59 floor swabs, and 36 stools from in-contact humans were collected and processed for Salmonella isolation. Isolates were tested for their susceptibility to 15 antimicrobials using Kirby-Bauer disk diffusion assay. Seventeen (28.8%) of the farms and 24 (2.9%) of the samples from poultry farms and 2.8% (1/36) of stool samples of humans in-contact with poultry were positive for Salmonella. Most of the isolates (n = 21) were recovered from fecal droppings of birds while the remaining isolates were recovered from floor swab samples (n = 2) and cloacal swab sample (n = 1). Only three Salmonella serovars: S. Haifa (n = 14, 56%), S. Anatum (n = 7; 28%), and S. Give (n = 4; 16%) were detected. Poultry farms in Adama town, large flock sized farms, and farms that used antimicrobials were significantly associated with the occurrence of Salmonella (p < .05). Twenty (80%) and 19 (76%) of Salmonella isolates were resistant to streptomycin and tetracycline, respectively. Nineteen (76%) of the isolates were resistant to two or more antimicrobials. Detection of multidrug-resistant strains of Salmonella in poultry farms suggests the need for detailed epidemiological and molecular studies to establish sources of acquisition of resistant Salmonella strains.

Methicillin-resistant Staphylococcus aureus (MRSA) isolated from chicken meat and giblets often produces staphylococcal enterotoxin B (SEB) in non-refrigerated raw chicken livers.

Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for several difficult-to-treat infections and staphylococcal food poisoning (SFP). This study was conducted to investigate the prevalence and enterotoxigenicity of MRSA in broiler chicken meat and giblets. A total of 5.5% (8/144) of the examined samples were contaminated with mecA positive/mecC negative MRSA, with staphylococcal counts of approximately 102 colony forming units (CFU)/g in breast, leg and gizzard samples and approximately 3.3 × 103 CFU/g in frozen liver samples. Most MRSA isolates (75%, 6/8) harboured the staphylococcal enterotoxin B (seb) gene. Reverse transcription-PCR (RT-PCR) showed that MRSA isolates initiated SEB production in experimentally contaminated chicken livers within 24 h of storage at temperatures over 8 °C. SEB was maximally produced at 24 °C when the MRSA counts reached 7.3 × 103 ± 1.2 × 103 CFU/g sample homogenate. The current study concludes that the main broiler chicken MRSA isolates in Egypt harbour the seb gene. To mitigate possible SEB production, especially in broiler chicken livers, a maximum "out of refrigeration" time limit should be implemented for cold chain poultry products.

Antimicrobial susceptibility, biofilm formation and genetic profiles of Escherichia coli isolated from retail chicken meat.

Brazil is the number one exporter of chicken meat, and this industry maintains constant microbiological vigilance. The objective of this study was to characterize the pathogenicity, antimicrobial resistance (AMR) and the profile of biofilm production of Escherchia coli strains isolated from raw refrigerated cuts of chicken meat sold in retail markets of the four largest poultry companies in Brazil. We collected 150 samples of chicken meat, in order to isolate E. coli and performed susceptibility tests (to amoxicillin associated with clavulanic acid, ceftiofur, enrofloxacin, gentamicin, and trimethoprim + sulfamethoxazole). In addition, the disc approximation test to detect extended spectrum beta-lactamases enzymes (ESBLs) producers was performed. E. coli ability to form biofilm was checked using polystyrene microplates. We also searched for ESBLs genes (blaCTY-M2, blaSHV-1, blaTEM-1, blaCTX-M2, blaOXA-1, blaPSE-1 and AmpC) and adhesion genes (sfa/foc, afa/draB, iha, hrla, fimC, tsh, papC, mat, cr1, felA, fimH and papG) in ESBL-E. coli producers and in those E. coli classified as strongly biofilm formers, respectively. The overall percentage of E. coli isolation was 58.66%, with brand A having the highest percentage (70%), followed by brands D, B and C (60, 53.3 and 50%, respectively). The highest resistance profile was observed for beta-lactams (39.5%), followed by sulfonamide associated to trimethoprim (36.9%) and polymyxin (33.4%). Of the isolates obtained, 77% were non-susceptible to at least one antimicrobial. Brand A showed the highest overall percentage of resistance with 95.23%, followed by brands C (80%), B (75%) and D (69.44%). Overall, 73.86% of the isolates were non susceptible to at least one antibiotic and 36.3% were multiresistants. A total of 17.04% of E. coli strains were identified as ESBLs producers and 70.44% were able to form biofilms (moderate-to-strong). The blaTEM-1 gene was the most prevalent (73.33%), followed by blaSHV-1 (46.66%) and blaCMY-2 (6%). Of the 31 strongly biofilm-forming strains, 26 (83.87%), 24 (77.41%) and 20 (64.51%) expressed fimC, papG and crl genes, respectively. Taken together, our results show that Brazilian chicken meat can be contaminated with E. coli that are non-susceptible to multiple antibiotics, able to form biofilm and showing a diverse repertoire of adhesins linked to pathogenicity depending on the brand evaluated.

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.

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.

Effects of climatic elements on Salmonella contamination in broiler chicken meat in Japan.

The effects of climatic elements on Salmonella contamination of chicken meat were investigated. Logistic regression analysis was performed to evaluate the association between Salmonella isolation, for 240 chicken samples purchased from March 2015 to February 2017, and climatic elements, over 65 days of chicken rearing. Salmonella was isolated from 143 samples (59.6%), and the most dominant serovars identified were Infantis (77/240, 32.1%) and Schwarzengrund (56/240, 23.3%). Previous studies have reported S. Schwarzengrund contamination of broiler chickens only in western Japan; however, in the present study, S. Schwarzengrund was also isolated from meat produced in eastern Japan-20% (12/60) in the C prefecture to 36.4% (8/22) in the Y prefecture-suggesting that S. Schwarzengrund-contaminated areas have expanded towards eastern Japan. Air temperature showed a significant negative association with S. Schwarzengrund isolation for chicken meat produced during periods with rising temperature (spring and summer) [odds ratio (OR), 0.894 to 0.935; P<0.01]. Moreover, the risk of S. Schwarzengrund contamination of chicken meat was higher during spring (OR, 3.951; P=0.008) and winter (OR, 4.071; P=0.006) than during summer. Effects of climatic elements and differences in contamination risk across seasons were not observed for any Salmonella serovars and only S. Infantis, which could be attributed to differences in transmission patterns and vehicles among Salmonella serovars. These findings are valuable for understanding the dynamics of S. Schwarzengrund dissemination in broiler farms.

Assessment of the microbiological quality and safety of marinated chicken products from Greek retail outlets.

The prevalence of three pathogens in marinated chicken products and the evaluation of their quality by microbiological and sensory analysis were assessed. Eighty (80) samples obtained from several meat retail markets in Greece were analyzed for the presence of Campylobacter spp., Salmonella and Listeria monocytogenes. Concerning Campylobacter, rep-PCR and species specific PCR were applied for the differentiation and identification of isolates, respectively. The samples were subsequently stored aerobically at 4 °C for 5 days. Microbiological analysis, sensory assessment and HPLC analysis were carried out for the evaluation of spoilage microorganisms, sensory quality and the presence of preservatives (potassium sorbate and sodium benzoate). Τhe prevalence of Campylobacter spp., Salmonella, and Listeria monocytogenes was 50%, 11% and 44%, respectively. In the case of Campylobacter, from a total of 40 isolates, 27 were identified as Campylobacter coli, 4 as Campylobacter jejuni, whereas the remaining 9 belonged to unidentified Campylobacter species. Pseudomonas spp. was the dominant spoilage microbial genus in 43% of the samples, while in 31% of them a co-dominance of Pseudomonas spp. and Brochothrix thermosphacta was observed. Total aerobic counts increased to 7.0 log CFU/g at the 1st, 2nd or 3rd day of storage in 71% of the samples, while sensory analysis showed that 80% of the samples were characterized as spoiled after 3, 4 or 5 days. The presence of preservatives was confirmed in 31% of the samples and slightly affected the microbiological profile. In conclusion, the obtained data demonstrated the occurrence of foodborne pathogens and allowed the acquisition of an overall view about the microbiological quality of marinated chicken products.

Prevalence and characteristics of Livestock-Associated Methicillin-Resistant Staphylococcus aureus (LA-MRSA) isolated from chicken meat in the province of Quebec, Canada.

This study was conducted to estimate the prevalence of Livestock-Associated Methicillin-Resistant Staphylococcus aureus (LA-MRSA) in retail chicken meat and broiler chickens from the Province of Quebec, Canada, and to characterize LA-MRSA isolates. A total of 309 chicken drumsticks and thighs were randomly selected in 2013 from 43 retail stores in the Monteregie. In addition, nasal swabs and caeca samples were collected in 2013-2014 from 200 broiler chickens of 38 different flocks. LA-MRSA was not detected in broiler chickens. Fifteen LA-MRSA isolates were recovered from four (1.3%) of the 309 chicken meat samples. Multi-Locus Sequence Typing (MLST) and SCCmec typing revealed two profiles (ST398-MRSA-V and ST8-MRSA-IVa), which were distinct using pulse-field gel electrophoresis (PFGE) and microarray (antimicrobial resistance and virulence genes) analyses. In addition to beta-lactam resistance, tetracycline and spectinomycin resistance was detected in all isolates from the 3 positive samples of the ST398 profile. Southern blot hybridization revealed that the resistance genes aad(D) and lnu(A), encoding resistances to aminoglycosides and lincosamides respectively, were located on plasmid. All isolates were able to produce biofilms, but biofilm production was not correlated with hld gene expression. Our results show the presence of two separate lineages of MRSA in retail chicken meat in Quebec, one of which is likely of human origin.