Prevalence of colistin-resistant Escherichia coli in foods and food-producing animals through the food chain: A worldwide systematic review and meta-analysis.

The purpose of this systematic review and meta-analysis was to summarize the available scientific evidence on the prevalence of colistin-resistant Escherichia coli strains isolated from foods and food-producing animals, the mobile colistin-resistant genes involved, and the impact of the associated variables. A systematic review was carried out in databases according to selection criteria and search strategies established a priori. Random-effect meta-analysis models were fitted to estimate the prevalence of colistin-resistant Escherichia coli and to identify the factors associated with the outcome. In general, 4.79% (95% CI: 3.98%-5.76%) of the food and food-producing animal samples harbored colistin-resistant Escherichia coli (total number of colistin-resistant Escherichia coli/total number of samples), while 5.70% (95% confidence interval: 4.97%-6.52%) of the E. coli strains isolated from food and food-producing animal samples harbored colistin resistance (total number of colistin-resistant Escherichia coli/total number of Escherichia coli isolated samples). The prevalence of colistin-resistant Escherichia coli increased over time (P < 0.001). On the other hand, 65.30% (95% confidence interval: 57.77%-72.14%) of colistin resistance was mediated by the mobile colistin resistance-1 gene. The mobile colistin resistance-1 gene prevalence did not show increases over time (P = 0.640). According to the findings, other allelic variants (mobile colistin resistance 2-10 genes) seem to have less impact on prevalence. A higher prevalence of colistin resistance was estimated in developing countries (P < 0.001), especially in samples (feces and intestinal content, meat, and viscera) derived from poultry and pigs (P < 0.001). The mobile colistin resistance-1 gene showed a global distribution with a high prevalence in most of the regions analyzed (>50%). The prevalence of colistin-resistant Escherichia coli and the mobile colistin resistance-1 gene has a strong impact on the entire food chain. The high prevalence estimated in the retail market represents a potential risk for consumers' health. There is an urgent need to implement based-evidence risk management measures under the "One Health" approach to guarantee public health, food safety, and a sustainable future.

Macroencapsulation of Limosilactobacillus reuteri DSPV002C as nutritional supplement for piglets: Storage stability and survival in gastrointestinal conditions.

The aim of this study was to evaluate the protective effect of the encapsulation of Limosilactobacillus reuteri DSPV002C in macrocapsules made from industrial materials during production, storage and under simulated gastrointestinal conditions in vitro and in vivo. The production of macrocapsules involved the evaluation of different wall materials (matrix), namely, gelatin and pregelatinized starch, different inoculums, matrix ratios, and diverse cryoprotectants (whey permeate and maltodextrin). The different macrocapsules were arranged in molds of similar size to pig pelleted food and lyophilized. Then, the viability of the macrocapsules was assessed over time during storage at different temperatures (freezing, refrigeration and room temperature) and atmospheres (vacuum and non-vaccum). The macrocapsules with 10% w/v gelatin+5% w/v pregelatinized starch, permeated (10%, w/v), with a 9:1 inoculum:matrix ratio (GS7.5P9), stored under freezing conditions and vacuum, exhibited the highest viability of L. reuteri DSPV002C (9.3 log CFU/cap until 210 d). Under simulated gastrointestinal conditions, the encapsulated inoculum showed less viability loss (0.58±0.09 log CFU/ml, 26.53%), compared to the free culture (1.56±0.16 log CFU/ml, 2.85%). Finally, by administering GS7.5P9 to pigs, the tolerance of the bacteria to the gastrointestinal environment was verified, with viable counts equal to or greater than 3.72 log CFU/g of fecal matter throughout the trial. In this study, a high-density carrier probiotic macrocapsule of L. reuteri DSPV002C was obtained, which displayed a long shelf life, a suitable shape to be included in pig feed and an adequate survival of viable cells at the site of action.

Lactic acid bacteria viability in different refrigerated food matrices: a systematic review and Meta­analysis.

The aim of this systematic review and meta-analysis was to determine which variables affect the viability of lactic acid bacteria (LAB) added to different types of refrigerated foods during the first 28 days. Scopus, ScienceDirect, PubMed and Cochrane Central Register of Reviews databases were searched from 1997 to April 2022. A total of 278 studies, which showed randomized and controlled experiments published in peer reviewed journals, were included. The viability of LAB in different moments during the storage process was synthesized as mean point estimate (MPE) via random-effects meta-analyses and the effect of multiple factors on the LAB´s viability was evaluated by multiple meta-regression. The meta-analysis showed that the decrease in LAB viability will be more abrupt the greater the initial dose. The physical structure of food may influence bacterial viability. Fruit was the type of product that most quickly lost viability. Co-culture of two or more species did not affect viability. Preservation methods had an unfavorable effect and prebiotics had a beneficial effect on bacterial viability. Viability was genus dependent. The data obtained in this study provide an overview of the factors to be taken into account for the design of new foods.

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.

Impact of farm-level strategies against thermotolerant Campylobacter in broiler chickens, using a quantitative risk assessment model and meta-analysis.

The aim of this study was to evaluate the effect of control strategies (probiotic supplementation and vaccination) at farm on thermotolerant Campylobacter infection due to the consumption of salad cross-contaminated from broiler meat. The broiler food chain was modelled considering the most common practices applied in Argentina (baseline model) and their effect on the prevalence and counts of Campylobacter. Probiotic supplementation and vaccination against Campylobacter spp. were included in different models to evaluate their effectiveness. The parameter distributions of each intervention were obtained based on a systematic review and meta-analysis previously described. The control measures applied at farms were evaluated considering their effectiveness in reducing both the prevalence and the count of thermotolerant Campylobacter in comparison with the baseline model estimation and expressed as relative change in campylobacteriosis risk. Additionally, the identification of the most important input parameters for the model was performed by sensitivity analysis. The model estimated a risk of campylobacteriosis per consumed serving of salad contaminated with poultry meat of 4.99 x 10-3 (95% CI: 6.12 x 10-6 -1.13 x 10-2 ), corresponding to an annual incidence risk estimated of 1,876,009 persons. Scenario analysis indicated that the application of vaccines against Campylobacter (probability of campylobacteriosis = 9.55 x 10-4 ; 95% CI: 5.31 x 10-4 -1.29 x 10-3 ) and the supplementation of broilers with probiotics (probability of campylobacteriosis = 1.32 x 10-3 ; 8.55 x 10-4 -1.69 x 10-3 ) can offer a modest reduction in risk estimates. The intervention efficacy was 80.86% and 73.54% for vaccination and probiotic supplementation, respectively. On-farm interventions were effective to mitigate the risk of campylobacteriosis.

Capacidad de Lactiplantibacillus plantarum LP5 para inhibir biopelículas de Campylobacter coli / Ability of Lactiplantibacillus plantarum LP5 to inhibit biofilms of Campylobacter coli

Resumen El objetivo de este estudio fue evaluar la capacidad inhibitoria de Lactiplantibacillus plantarum LP5 frente a Campylobacter coli en ensayos de formación de biopelículas in vitro y exclusión competitiva. La formación de biopelículas por C. coli NCTC11366, C. coli DSPV458, C. coli DSPV541 y C. coli DSPV570 fue evaluada mediante medición de DO. La capacidad inhibitoria de L. plantarum LP5 frente a C. coli fue evaluada sobre discos de vidrio, nailon y aluminio. Sobre una biopelícula de L. plantarum se adicionó C. coli para cuantificar el efecto inhibidor de L. plantarum LP5 sobre el patógeno. Las cuatro cepas de C. coli fueron clasificadas como moderadas formadoras de biopelículas. El ensayo de exclusión competitiva mostró que la formación de biopelículas de las cepas de C. coli en todos los materiales fue significativamente mayor que la formación de biopelículas de cada patógeno en presencia de biopelículas de L. plantarum LP5. Si bien es necesario realizar más pruebas para confirmar la capacidad de supervivencia de C. coli en ambientes hostiles hasta llegar al huésped, este estudio permitiría avanzar en el esclarecimiento de su comportamiento mediante la formación de biopelículas.

Abstract The objective of this study was to evaluate the inhibitory capacity of Lactiplantibacillus plantarum LP5 against Campylobacter coli in in vitro biofilm formation and competitive exclusion assays. Biofilm formation by C. coli NCTC11366, C. coli DSPV458, C. coli DSPV541 and C. coli DSPV570 was evaluated by OD measurement. The inhibitory capacity of L. plantarum LP5 against C. coli was evaluated on glass, nylon and aluminium discs, added with L. plantarum and incubated at 37°C for 72 h. C. coli was added to each washed well. The plates were incubated at 42°C for 72 h in microaerophilic conditions and the biofilms were detached for quantification. The four strains of C. coli were classified as moderate biofilm former. The competitive exclusion test showed that the biofilm formation of the C. coli strains in all materials was significantly higher than the biofilm formation of each pathogen in the presence of L. plantarum LP5 biofilms. Although it is necessary to carry out more tests to confirm the ability of C. coli to survive in hostile environments until reaching the host, this study would allow progress in the elucidation of its behaviour through the formation of biofilms.

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.

Development and storage studies of high density macrocapsules containing Lactobacillus spp. strains as nutritional supplement in young calves.

The aim of this study was to evaluate different production methodologies of probiotic macrocapsules with high bacterial densities destined to lactating calves. Three types of capsules containing Lactobacillus casei DSPV318T and Lactobacillus plantarum DSPV354T were prepared from an overnight culture in whey medium: (1) mixing the culture with calcium alginate and then, reincubating the capsules in whey (RC); (2) concentrating the biomass by centrifugation and mixing the pellet with calcium alginate (CC) at different concentrations with respect to the initial culture (5X and 12.5X); (3) CC with cryoprotectants: whey permeate (Per) and glycerol (Gly). Chitosan coating was evaluated. Capsules were freeze-dried and viability was assessed before freezing, after freeze-drying and every two weeks for 84 days of storage at room temperature, 4°C and -20°C. CC showed higher cell densities than RC. Storage temperature affected viability: greater viability at lower temperature. Moreover, the effect of temperature was influenced by other factors, such as capsule coating, culture neutralization and cryoprotectants. Coating improved viability at room temperature; however no effect was observed at 4°C. Culture neutralization allowed greater survival during storage. Cryoprotectants improved viability during freezing, but they also generated a positive or negative effect depending on storage temperature. The best results were: at refrigeration Gly12.5X exhibited counts above 109CFU/capsule until day 70 and Per12.5X until day 56 of storage and at -20°C Gly12.5X showed counts above 109CFU/capsule until the end of the study (84 days). A 109CFU capsule is the daily dose per calf which would facilitate the administration of this probiotic inoculum to field animals.

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.