Applicability of the probiotic Lactiplantibacillus plantarum BFL as an adjunct culture in a dry fermented sausage.

The addition of probiotic bacteria to a meat batter allows the development of functional fermented sausages. The aim of this work was to study the effect of microencapsulated Lactiplantibacillus plantarum BFL (EP) and as free cells (FP) on microbiological, physicochemical, and sensory parameters of fermented sausages during the drying stage and on the product ready for consumption. The microencapsulation of L. plantarum BFL did not improve its viability during the drying stage. In addition, sausages inoculated with L. plantarum BFL (FP and EP) caused lower residual nitrites values, pH values and Escherichia coli counts than the Control (C). However, only the presence of free cells of L. plantarum BFL (FP) caused a decrease in the Enterobacteriaceae and mannitol salt-positive Staphylococcus counts. In the sensory analysis, no significant differences were found in the acceptability of the different sausages. However, the acidity in probiotic sausages (FP and EP) was an attribute that consumers highlighted. The probiotic L. plantarum BFL could adapt and survive at high doses in the matrix of an industrial fermented sausage. Therefore, its use could represent a strategy both for biocontrol of pathogens and for the development of functional meat products.

Prevalence and antimicrobial resistance of Campylobacter jejuni and C. coli identified in a slaughterhouse in Argentina.

The aim of this study was to evaluate the percentage of Campylobacter (C. jejuni and C. coli) from samples collected at the slaughterhouse to describe the prevalence of resistance to selected antimicrobials, and to characterize the genetic determinants. In total, from 333 samples analyzed, 31% were positive for Campylobacter. More positive samples were detected before the chiller (46%) than after the chiller (16%). C. coli (59%) was more prevalent than C. jejuni (41%). Antimicrobial resistance differences between C. jejuni and C. coli were found (p < 0.001). Multidrug resistance was found in 72% of C. coli isolates and 69% of C. jejuni isolates (p < 0.001). Most C. jejuni isolates (57%) had the three genes of the cmeABC efflux pump. The tet(O) gene and resistance-associated point mutations within both the gyrA and 23S rRNA genes were detected in 100% of C. coli isolates. On the other hand, C. jejuni only had more prevalence of the bla OXA-61 gene than C. coli (p < 0.001), and most of the C. jejuni isolates (70-80%) had the tet(O) and gyrA point mutation. These results could contribute to knowledge about the status of thermotolerant Campylobacter resistant to antimicrobials isolated from food animals in Argentina and to develop an antimicrobial resistance surveillance system.

Prevalence, genotypic diversity and detection of virulence genes in thermotolerant Campylobacter at different stages of the poultry meat supply chain.

Thermotolerant Campylobacter is the leading bacterial cause of foodborne illness in humans worldwide. The objectives of this study were to estimate prevalence and to identify and characterize potential sources of thermotolerant Campylobacter contamination in broilers on farms and at the slaughterhouse; to evaluate the clonal relationship among thermotolerant Campylobacter isolates from different stages of the broiler meat supply chain, and to analyze the presence of virulence genes in different sources of thermotolerant Campylobacter. A total of 1210 samples were collected from three broiler meat supply chains in Santa Fe, Argentina. At the farms, the sampling collection included broilers one week prior to slaughter, wild-living birds, domestic dogs, wild rodents, farm workers' boots, litter, feed, drinking water, flies, and darkling beetles (Alphitobius diaperinus). At the slaughtering line, the samples taken were from the evisceration zone (broiler cecum, working surfaces, evisceration knives and workers' hands), from the chiller zone (surfaces and direct supply water) and from the packing zone (work surfaces, workers' hands and broiler carcasses). The samples taken along each supply chain were in the same batch. The isolates obtained were identified to the species level (C. jejuni and C. coli) by multiplex PCR and were analyzed using pulsed-field gel electrophoresis to compare different profiles according to the source. Finally, the presence of 11 virulence genes was examined (cadF, cdtA, cdtB, cdtC, ciaB, flaA, flhA, iam, wlaN, virB11, racR). From 254 isolates, 128 (50.4%) were Campylobacter jejuni and 126 (49.6%) Campylobacter coli. C. jejuni was the species most prevalent in farm and C. coli the species most prevalent at the slaughterhouse. We detected thermotolerant Campylobacter in samples of wild birds, darkling beetles, farm workers' boots, flies and litter. At the slaughterhouse, the prevalence varied along the process line. By analyzing PFGE results, C. jejuni showed 21 profiles with three predominant genotypes, while C. coli showed 14 profiles with four predominant genotypes. A high genotype diversity was found; however, relationships between isolates from different stages of the broiler meat chain, between broiler and potential sources of thermotolerant Campylobacter contamination and between strains in the farm and in the slaughterhouse were detected. Furthermore, there was evidence of cross-contamination at the slaughterhouse. FlaA, flhA genes were detected in all strains, and the third most prevalent virulence gene was cadF. Only those strains obtained from flies, wild-living birds and broiler carcass samples harbored 10 of 11 pathogenic genes. The prevalence of some pathogenic genes between C. jejuni and C. coli was different. This evidence should contribute the scientific basis to implement risk management measures in public health.

Meta-analysis of the prevalence of thermotolerant Campylobacter in food-producing animals worldwide.

The objective of this meta-analysis was to summarize available information on the prevalence of thermotolerant Campylobacter (TC) in different food-producing animals worldwide. Databases (i.e., PubMed, ScienceDirect, Scopus) were searched from 1980 to 2017 unrestricted by language. The inclusion criteria were as follows: prevalence or incidence studies, published in peer-reviewed journals, and they must have reported the total number of animal samples studied and the number of samples that were positive for the presence of TC. When the identification of Campylobacter species was available, this information was included in the analysis. Multilevel random-effect meta-analysis models were fitted to estimate mean occurrence rate of TC and to compare them among different factors potentially associated with the outcome. The mean occurrence rate of TC in food-producing animals was 0.424 (95% CI: 0.394-0.455), and the mean occurrence rate of Campylobacter jejuni and Campylobacter coli were 0.214 and 0.133, respectively. Pigs and poultry showed the highest prevalence of TC; however, there were differences in the prevalence of each Campylobacter species. Campylobacter jejuni was observed in broilers (0.322; 95% CI: 0.273-0.377) and hens (0.395; 95% CI: 0.265-0.542), while C. coli was restricted essentially in pigs (0.553; 95% CI: 0.541-0.650). The prevalence of C. jejuni in intensively bred cattle was higher (0.302; 95% CI: 0.227-0.389) than the prevalence in extensively bred cattle (0.172; 95% CI: 0.119-0.242) while the prevalence of C. coli was similar (0.051; 95% CI: 0.028-0.091 vs. 0.050; 95% CI: 0.027-0.091) in both production systems. Agar with or without blood used for the isolation of TC did not affect the prevalence observed. The method of species identification did not seem to generate differences in the prevalence of Campylobacter species. The prevalence of Campylobacter in primary food production has a strong impact on the entire agri-food chain. National authorities must monitor the situation with the aim to establish the appropriate risk management measures.

Antimicrobial Resistance of Thermotolerant Campylobacter Species Isolated from Humans, Food-Producing Animals, and Products of Animal Origin: A Worldwide Meta-Analysis.

The objective of this meta-analysis was to summarize available information on the prevalence of antimicrobial-resistant Campylobacter species in humans, food-producing animals, and products of animal origin. A number of multilevel random-effect meta-analysis models were fitted to estimate mean occurrence rate of antimicrobial-resistant thermotolerant Campylobacter and to compare them throughout the years and among the species, food-producing animals (i.e., bovine, pigs, broilers, hen, goat, and sheep), country of origin, sample type, methodology to determine the antimicrobial susceptibility, and the species of Campylobacter. Among the considered antibiotics, thermotolerant Campylobacter showed the highest resistance to tetracycline (pool estimate [PE] = 0.493; 95% CI 0.466-0.519), nalidixic acid (PE = 0.385; 95% CI 0.348-0.423), and ciprofloxacin (PE = 0.376; 95% CI 0.339-0.415). In general, the prevalence of antimicrobial-resistant Campylobacter spp. was higher in hen, broilers, and swine. Campylobacter coli showed a higher prevalence of antimicrobial resistance than Campylobacter jejuni. Independent of the antimicrobial evaluated, the disk diffusion method showed higher prevalence of antimicrobial-resistant Campylobacter than the methods based on the minimum inhibitory concentration estimation. The meta-analysis showed that the prevalence of antimicrobial-resistant Campylobacter is relevant essentially in foods derived from hens and broilers, and it was observed worldwide. The prevalence of this pathogen is of public health importance and the increase in the prevalence of Campylobacter strains resistant to the antimicrobial of choice worsens the situation, hence, national authorities must monitor the situation in each country with the aim to establish the appropriate risk management measures.

Genetic diversity of thermotolerant Campylobacter spp. isolates from different stages of the poultry meat supply chain in Argentina.

The objective of this study was to investigate a clonal relationship among thermotolerant Campylobacter spp. isolates from different stages of the poultry meat supply chain in Argentina. A total of 128 thermotolerant Campylobacter spp. (89 C. jejuni and 39 C. coli) isolates from six poultry meat chains were examined. These isolates were from: a) hens from breeder flocks, b) chickens on the farm (at ages 1 wk and 5 wk), c) chicken carcasses in the slaughterhouse, and d) chicken carcasses in the retail market. Chickens sampled along each food chain were from the same batch. Campylobacter spp. isolates were analyzed using pulsed-field gel electrophoresis to compare different profiles according to the source. Clustering of C. jejuni isolates resulted in 17 profiles, with four predominant genotypes and many small profiles with just a few isolates or unique patterns, showing a very high degree of heterogeneity among the C. jejuni isolates. Some clusters included isolates from different stages within the same chain, which would indicate a spread of strains along the same poultry meat chain. Moreover, twenty-two strains of C. coli clustered in seven groups and the remaining 17 isolates exhibited unique profiles. Evidence for transmission of thermotolerant Campylobacter spp. through the food chain and cross contamination in the slaughterhouses were obtained. This collective evidence should be considered as the scientific basis to implement risk management measures to protect the public health.

[Campylobacter jejuni O:19 serotype in Argentine poultry meat supply chain]. / Presencia de serotipos de Campylobacter jejuni O:19 en la cadena avícola Argentina.

Thermotolerant species of Campylobacter have been focus of attention in the last years because they are the major agent causing zoonotic foodborne diseases. In addition, Campylobacter jejuni O:19 serotype was associated with Guillain Barré syndrome. The aim of this study was to determine the proportion of C. jejuni O:19 serotype isolated at different stages of three poultry meat supply chain in Santa Fe, Argentina. The analysis showed that 18% of isolated C. jejuni belong to serotype O:19. It was also determined that the presence of these strains is given in almost all production stages. These results reflect a significant risk to public health of consumers. Epidemiological studies of Campylobacter should be considered to establish a risk manager policy.