Characterization of Extended Spectrum Cephalosporin-Resistant Escherichia coli Strains Isolated from Raw Poultry Carcasses in Catering Services in Northern Greece.

Antimicrobial resistance is considered a topic of utmost interest under the concept of "One Health", having severe implications in both human and veterinary medicine. Among the antibiotic-resistant bacteria, gram-negative bacteria, especially those belonging to the order of Enterobacterales (such as Escherichia coli), hold a prominent position in terms of both virulence and possessing/disseminating antimicrobial resistance (AMR) traits. The aim of this study was to examine the presence of extended-spectrum ß-lactamase producing E. coli isolates in raw poultry carcasses collected from a university club. Five hundred raw poultry skin samples were collected from the Aristotle University of Thessaloniki (AUTh) club in Thessaloniki, Greece. A total of 64% of the samples were positive for the presence of extended-spectrum ß-lactamase (ESBL)-producing E. coli. The isolates were further examined for their susceptibility to selected antibiotics by the disc diffusion method and were characterized as true ESBL, as producing class C cephalosporinases (AmpC) or "of unknown etiology" by the combination disc test. The 86 of the 120 isolates (71.67%) were classified as true ESBL, 24 (20.00%) as AmpC, and 10 (8.33%) as "of unknown etiology". The isolates were screened for the occurrence of ß-lactamase genes (blaTEM, blaCTX-M, blaSHV, and blaOXA). Thirty-six isolates (32 ESBL- and 4 AmpC-phenotype) harbored both blaTEM and blaCTX-M genes, twenty-two isolates (among which 19 ESBL-phenotype and 2 AmpC-phenotype) harbored blaCTX-M only, whereas twenty-six (14 ESBL- and 12 AmpC-phenotype) isolates harbored blaTEM alone. No isolate harboring blaSHV or blaOXA was detected. The results demonstrate the existence of E. coli isolates producing extended-spectrum ß-lactamases in poultry carcasses from Greece, pausing a risk for antibiotic resistance transfer to humans.

Livestock microbial landscape patterns: Retail poultry microbiomes significantly vary by region and season.

Microbes play key roles in animal welfare and food safety but there is little understanding of whether microbiomes associated with livestock vary in space and time. Here we analysed the bacteria associated with the carcasses of the same breed of 28 poultry broiler flocks at different stages of processing across two climatically similar UK regions over two seasons with 16S metabarcode DNA sequencing. Numbers of taxa types did not differ by region, but did by season (P = 1.2 × 10-19), and numbers increased with factory processing, especially in summer. There was also a significant (P < 1 × 10-4) difference in the presences and abundances of taxa types by season, region and factory processing stage, and the signal for seasonal and regional differences remained highly significant on final retail products. This study therefore revealed that both season and region influence the types and abundances of taxa on retail poultry products. That poultry microbiomes differ in space and time should be considered when testing the efficacy of microbial management interventions designed to increase animal welfare and food safety: these may have differential effects on livestock depending on location and timing.

Serotype diversity and slaughterhouse-level risk factors related to Salmonella contamination on poultry carcasses in Algiers.

INTRODUCTION: In Algeria, the latest studies on Salmonella demonstrated warning contamination rates in farms and slaughterhouses. This pathogen can contaminate poultry meat and put humans at risk especially that such product is nowadays widely consumed. METHODOLOGY: a cross-sectional study was conducted in Algiers to evaluate prevalence, determine serotypes and quantify risk for Salmonella contamination in broiler chickens and turkeys at the post-chill stage of slaughter process. RESULTS: batch prevalence was 63.1% for chickens and 34.9% for turkeys. Eleven serotypes were isolated from chickens and five from turkeys. The most predominant at both sample and batch levels was S. Kentucky either in chicken (65.1%) or in turkey carcasses (63.2%). Univariate analysis screened 3 variables for chickens and 5 variables for turkeys. Final multivariate regression models provided one potential risk factor for Salmonella contamination in each poultry species. Presence of less than 6 broilers simultaneously in the traditional scalding tank of small scale slaughterhouses had a significantly reduced contamination risk (OR = 0.31; p < 0.05). Slaughtering turkeys in sites processing only this specie than in mixed poultry slaughterhouses increased significantly the contamination probability (OR = 4.44; p < 0.05). CONCLUSIONS: Our study indicates a high prevalence of Salmonella-contaminated poultry carcass with wide diversity of serotypes. Moreover, two potential risk factors identified for the first time in Algeria are found to be associated with the lack in hygienic management on production sites. A real threat for consumers exists highlighting the imperative need for improved safety throughout the local poultry meat supply chain.

Prevalência e susceptibilidade antimicrobiana de sorotipos de Salmonella spp. isolados de frangos vivos e carcaças no estado do Rio de Janeiro / Prevalence and antimicrobial susceptibility of Salmonella spp. serotypes in broiler chickens and carcasses in the State of Rio de Janeiro, Brazil

A presença de Salmonella spp. em produtos de origem avícola e seus subprodutos se mostra um grande desafio para a produção comercial. Dados de prevalência, dos sorotipos circulantes e do perfil de susceptibilidade antimicrobiana de cepas de Salmonella spp. no Estado do Rio de Janeiro são escassos. Portanto, objetivou-se detectar a presença Salmonella spp. em frangos vivos e carcaças em matadouros do Estados do Rio de Janeiro, identificar os sorotipos e avaliar a susceptibilidade antimicrobiana dessas cepas para fluoroquinolonas e betalactâmicos. Foram coletadas 60 amostras cloacais de frangos vivos e 60 amostras de carcaça de seis matadouros sob Inspeção Estadual (SIE). Os isolados foram sorotipificados e testados frente a oito antimicrobianos: enrofloxacina, ciprofloxacina, norfloxacina, cefalotina, ceftiofur, cefotaxima, amoxicilina/ácido clavulânico e ampicilina pelo método de difusão em disco. Os resultados mostraram uma prevalência de Salmonella spp. de 1,66% (1/60) em amostras de suabe de cloaca e de 26,66% (16/60) em carcaças. Em amostras de suabe de cloaca, somente o sorotipo Senftenberg (1,66%) foi isolado. No total, foram isolados sete sorotipos diferentes nas carcaças: Senftenberg (15%) o mais frequente, seguido por Mbandaka (8,3%), Schwarzengrund (3,3%), Cerro (3,3%), Ohio (3,3%), Minnesota (1,66%) e Tennessee (1,66%). Em relação à susceptibilidade antimicrobiana, 29 (87,87%) isolados foram sensíveis a todos os antimicrobianos testados e 4 (12,12%) isolados foram resistentes a pelo menos três antimicrobianos betalactâmicos ou mais. Não foi observada resistência às fluoroquinolonas. Os resultados encontrados demonstram uma prevalência de Salmonella spp. acima da esperada em matadouros do Estado do Rio de Janeiro, além da presença de vários sorotipos de Salmonella spp. A resistência encontrada para betalactâmicos alerta para a disseminação dessas cepas pela cadeia alimentar.(AU)

The presence of Salmonella spp. in poultry products and their by-products is a major challenge for commercial production. Data about the prevalence, the circulating serotypes and the antimicrobial susceptibility profile of Salmonella spp. strains in the State of Rio de Janeiro are scarce. Therefore, the aim of this study was to detect the presence of Salmonella spp. in live chickens and carcasses in slaughterhouses of the State of Rio de Janeiro, to identify the serotypes and to evaluate the antimicrobial susceptibility of these strains for fluoroquinolones and beta-lactams. Sixty cloacal swabs samples from broiler chickens and sixty samples of carcasses from six slaughterhouses under State Inspection were collected. The isolates were serotyped and resistance was tested to eight antimicrobials: enrofloxacin, ciprofloxacin, norfloxacin, cephalothin, ceftiofur, cefotaxime, amoxicillin/clavulanic acid and ampicillin by disc diffusion method. The results showed a prevalence of Salmonella spp. of 1.66% (1/60) in cloacal swabs samples and 26.66% (16/60) in carcasses. In cloacal swabs sample only Senftenberg (1.66%) serotype was isolated. In total, seven different serotypes were obtained from carcasses: Senftenberg (15%), followed by Mbandaka (8.3%), Schwarzengrund (3.3%), Cerro (3.3%), Ohio (3.3%), Minnesota (1.66%) and Tennessee (1.66%). Regarding antimicrobial susceptibility, 29 (87.87%) isolates were sensitive to all antimicrobials tested and 4 (12.12%) isolates were resistant to three or more beta-lactams antimicrobials. No susceptibility to fluoroquinolones was observed. These results showed a prevalence of Salmonella spp. higher than expected in slaughterhouses in the State of Rio de Janeiro, besides the presence of several serotypes of Salmonella spp. The resistance found for beta-lactams alerts to the spread of these strains through the food chain.(AU)

Effect of Substrate on Identification of Microbial Communities in Poultry Carcass Composting and Microorganisms Associated with Poultry Carcass Decomposition.

Three composting systems, which consisted of different ratios of chicken manure, sawdust, and poultry carcasses, were used to investigate the effect of substrate on the identification of microbial communities and microorganisms associated with poultry carcass decomposition by characterizing the microbial communities and physicochemical parameters. The physicochemical and Miseq Illumina sequencing results showed the composition of substrate had a significant effect on the identification and metabolic capabilities of microbial communities in decomposting process. Poultry carcasses might be the potential driver for the identification of bacterial communities in poultry carcass composting, whereas the initial C/N ratio may mainly contribute to the diversity of fungal communities and the similar dominant microbial communities in treatments. Poultry carcasses and initial C/N ratio could respectively affect the composition and abundance of microorganisms associated with the decomposition of poultry carcasses. Understanding the potential composting driver could allow development of an efficient carcass degradation system.

Predicting Salmonella Typhimurium reductions in poultry ground carcasses.

To improve understanding of Salmonella Typhimurium LT2 inactivation in ground poultry carcasses, a series of experiments were carried out at multiple temperatures. Subsequently, a non-linear model was developed to predict Salmonella inactivation at composting and low rendering temperatures. The Salmonella inactivation study was conducted using bench-top experiments at 38, 48, 55, 62.5, 70, and 78°C in mixed and non-mixed reactors using ground poultry carcasses as a feedstock. Subsequently, these observations were used for developing a non-linear model. The model predictions were compared with the observations of a different set of experiments. The comparisons among predictions and observations showed that the model predictions are reasonable and can be useful to determine the time required for Salmonella inactivation in poultry carcasses at multiple temperatures. Results showed that at composting conditions, when temperature varies between 48 and 62.5°C, Salmonella survival can prolong between 10,000 and 25,000 min (7 to 17 d). If ambient temperature is maintained at low temperature rendering range (70 to 78°C), then Salmonella survival can last for 90 to 120 minutes. We anticipate that this study will help in improving the existing understanding of Salmonella survival in poultry carcasses.

Assessing Salmonella typhimurium persistence in poultry carcasses under multiple thermal conditions consistent with composting and wet rendering.

Mitigation of Salmonella associated with poultry carcasses is primarily accomplished by rendering or carcass composting. While rendering temperatures and pressures are well established for pathogen inactivation in poultry carcasses, parameters controlling composting processes are less defined in part because multiple conditions and procedures are utilized. Consequently, limited knowledge exists describing the impacts of composting with varying temperature and mixing protocols with respect to the inactivation of Salmonella in poultry carcasses. To improve the existing knowledge of Salmonella survival in poultry carcasses, inactivation of Salmonella enterica serovar Typhimurium (ST) LT2 was investigated. The impacts of various composting temperatures (55, 62.5°C) and low-rendering (i.e., pasteurization) temperatures (70, 78°C) on Salmonella inactivation were tested in a bench-top setting using a ground carcass slurry and whole birds under mixed and non-mixed conditions. Results showed that the ground carcass slurry and the whole carcass exposed to temperatures consistent with composting had no detectable Salmonella after 110 h with a level of detection of one CFU/mL of ground carcass slurry and one CFU/g of whole carcasses, respectively. In addition, grinding of carcasses as opposed to whole carcasses was more predictable with respect to Salmonella heat inactivation. Furthermore, results showed that constant mixing decreased the overall time required to eliminate Salmonella under composting and low-rendering temperatures.

Poultry litter of wood shavings and/or sugarcane bagasse: animal performance and bed quality / Cama de viruta de madera y/o bagazo de caña de azúcar para pollo: desempeño animal y calidad de la cama / Cama composta por maravalha e/ou bagaço de cana de açúcar para frangos: desempenho animal e qualidade da cama

Background: litter provides comfort to animals while improving productive performance and carcass quality. Objective: this study evaluated broiler performance, incidence of contact dermatitis, and quality of poultry litter of wood shavings and/or sugarcane bagasse (SB) after five consecutive flocks. Methods: two thousand birds were raised in five consecutive flocks composed of 400 chicks each. The experiment was set in a completely randomized design with five treatments and four repetitions. The treatments consisted of substituting wood shavings with different levels of SB (0, 25, 50, 75, and 100%). Results: litter density, pH, dry matter content, total nitrogen, and phosphorus did not vary between treatments. However, substituting wood shavings with SB increased moisture retention capacity of the litter and ammonia production. Neither carcass lesions nor productive performance, with the exception of weight gain and feed conversion ratio in the fifth flock, differed between treatments. Conclusions: substituting 75% of wood shavings with SB maintained litter quality and improved poultry performance.

Antecedentes: la cama propicia el confort y mejora el desempeño productivo y la calidad de la canal de los pollos de engorde. Objetivo: el objetivo del presente estudio fue evaluar el desempeño productivo de pollos de engorde, la incidencia de dermatitis de contacto en la canal y la calidad de la cama de pollo compuesta de viruta de madera y/o bagazo de caña de azúcar (SB), usada en cinco lotes consecutivos. Métodos: dos mil aves fueron criadas en cinco lotes consecutivos, cada uno constituido por 400 pollitos, en un diseño completamente aleatorizado con cinco tratamientos y cuatro repeticiones. Los tratamientos consistieron en diferentes niveles de sustitución de viruta de madera con SB (0, 25, 50, 75 y 100%). Resultados: la densidad, pH y contenidos de materia seca, nitrógeno total y fósforo no variaron entre los tratamientos. Sin embargo, sustituir viruta de madera con SB aumentó la capacidad de retención de humedad y la cantidad de amoníaco volatilizado en el galpón. Ni las lesiones en la canal y ni el desempeño productivo, con excepción de la ganancia de peso y conversión alimentaria en el quinto lote, difirieron entre los tratamientos. Conclusiones: sustituir 75% de la viruta de madera con SB mantuvo la calidad de la cama y mejoró el desempeño productivo de las aves.

Antecedentes: cama de frango propicia conforto e melhora o desempenho produtivo e a qualidade da carcaça das aves. Objetivo: este estudo avaliou o desempenho produtivo de frangos de corte, a incidência de dermatite de contato na carcaça e a qualidade da cama de frango composta de maravalha e/ou bagaço de cana (SB), usada em cinco lotes consecutivos. Métodos: duas mil aves foram criadas em cinco lotes consecutivos, compostos de 400 pintinhos cada, em delineamento inteiramente casualizado com cinco tratamentos e quatro repetições. Os tratamentos consistiram de diferentes níveis de substituição de maravalha com BC (0, 25, 50, 75 e 100%). Resultados: densidade, pH e teores de matéria seca, nitrogênio total e fósforo não variaram entre os tratamentos. Entretanto, substituir maravalha com SB aumentou a capacidade de retenção de umidade e a quantidade de amônia volatilizada no galpão. Nem as lesões na carcaça e nem o desempenho produtivo, com exceção do ganho de peso e conversão alimentar no quinto lote, diferiram entre os tratamentos. Conclusões: substituir 75% da maravalha com SB manteve a qualidade da cama e melhorou o desempenho produtivo das aves.

Rapid identification of Campylobacter jejuni from poultry carcasses and slaughtering environment samples by real-time PCR.

The objective of this study was to develop a real-time PCR assay for rapid identification of Campylobacter jejuni and to apply the method in analyzing samples from poultry processing. A C. jejuni-specific primer set targeting a portion of the C. jejuni hippuricase gene was developed. The specificity of the newly designed primer pair was verified using 5 C. jejuni strains and 20 other bacterial strains. Sensitivity was determined to be as low as 1 genome copy per reaction. A total of 73 samples were collected at different sites along the processing line during 2 visits to a poultry slaughterhouse and were examined by direct plating onto modified charcoal cefoperazone deoxycholate agar or after enrichment in Bolton broth followed by plating on modified charcoal cefoperazone deoxycholate agar. The newly developed real-time PCR assay was used to identify the presumptive colonies as belonging to C. jejuni. A real-time PCR assay targeting 16S ribosomal RNA was also applied to determine Campylobacter spp. prevalence. Results from the real-time PCR analysis indicated considerable variability in Campylobacter contamination, with incidence rates of 72.7 and 27.6% for sampling days A and B, respectively. Campylobacter was isolated from 100% of prescalded and preeviscerated carcasses on sampling day A. In contrast, on sampling day B, the highest number of Campylobacter-positive carcasses was recovered after evisceration (60%). The chilling process significantly reduced (P < 0.05) Campylobacter population, but the percentage of positive samples on sampling day A increased to 80%. All samples collected from the processing environment, except scalding tank 3 and the prechiller and chiller tanks, were 100% positive on day A, whereas no campylobacters were isolated from machinery on sampling day B. Our results revealed the widespread of C. jejuni in poultry processing and proved that the newly developed real-time PCR assay is a simple, specific, and inexpensive method for rapid C. jejuni identification. The newly developed PCR method can be easily used in laboratories for reliable and unambiguous identification of C. jejuni in poultry samples.