Salmonella Control in Fattening Pigs through the Use of Esterified Formic Acid in Drinking Water Shortly before Slaughter.

The presence of Salmonella in pig feces is a major source of abattoir and carcass contamination, and one of the main sources of human salmonellosis. This study assessed whether using a form of esterified formic acid (30% formic acid) in drinking water (10 kg/1000 L) 5 days before slaughter could be a helpful strategy to mitigate this public health issue. Thus, 240 pigs from three Salmonella-positive commercial fattening farms were selected. From each farm, 40 pigs were allocated to a control group (CG) and 40 to a treatment group (TG). At the abattoir, fecal samples from both groups were collected for Salmonella detection (ISO 6579-1:2017) and quantification (ISO/TS 6579-2:2012). Salmonella was present in 35% (95% IC = 29.24-41.23) of the samples collected. The prevalence was significantly higher in the CG than in the TG (50% vs. 20%; p < 0.001). In all farms, the TG showed a lower percentage of shedders than the CG. A random-effects logistic model showed that the odds of shedding Salmonella were 5.63 times higher (95% CI = 2.92-10.8) for the CG than for the TG. Thus, the proportion of pigs shedding Salmonella that was prevented in the TG due to the use of this form of organic acid was 82.2%. In addition, a Chi-squared analysis for trends showed that the higher the Salmonella count, the higher the odds of the sample belonging to the CG. These results suggest that adding this type of acid to drinking water 5 days before slaughter could reduce the proportion of Salmonella-shedding pigs and the Salmonella loads in the guts of shedder pigs.

Building a predictive model for assessing the risk of Salmonella shedding at slaughter in fattening pigs.

Salmonellosis continues to be a major cause of foodborne outbreaks worldwide, and pigs are one of the main sources of human infection. Salmonella pork contamination is a major concern for abattoirs and is related to the presence of Salmonella in pigs' feces at slaughter. Being able to predict the risk of Salmonella shedding in pigs arriving at the slaughterhouse could help mitigate abattoir and carcass contamination. For this purpose, 30 batches of 50 pigs each were selected from 30 different fattening units. The pigs were tagged and bled for the detection of antibodies against Salmonella approximately one month before slaughter. Pooled floor fecal samples were also collected from 10 pens per unit for Salmonella detection, and a questionnaire on biosecurity was administered to each farm. At the abattoir, colon content was collected from each tagged pig for the Salmonella shedding assessment. A predictive model for Salmonella shedding at slaughter was built with two-third of the pigs by employing random-effects logistic regression analysis, with Salmonella shedding as the dependent variable and pig serology and other farm/environmental characteristics as the independent variables. The model included farm as the grouping factor. Data from the remaining one-third of the pigs were used for model validation. Out of 1,500 pigs initially selected, 1,341 were identified at the abattoir and analyzed. Salmonella was detected in 13 (43.3%; 95%CI = 27.4-60.8) of the fattening units. The mean batch seroprevalence (cut-off OD% ≥40) among the fattening units was 31.7% (95%CI = 21.8-41.0), and a total of 316 pigs (23.6%; 95%CI = 21.4-25.9) shed Salmonella at slaughter. The model predicted reasonably well (Area under the curve = 0.76; P < 0.05) whether a pig would shed Salmonella at slaughter, with estimates of sensitivity and specificity at 71.6% and 73.6%, respectively. Serology, the percentage of Salmonella-positive pens on the farm, and the internal biosecurity score were significantly associated (P < 0.05) with Salmonella shedding at the abattoir, and several scenarios were observed by the model. The study highlighted that although serology may be helpful for identifying batches of pigs at risk of shedding Salmonella upon their arrival at the abattoir, it may not be necessary in some scenarios.

Salmonella Shedding in Slaughter Pigs and the Use of Esterified Formic Acid in the Drinking Water as a Potential Abattoir-Based Mitigation Measure.

Pigs shedding Salmonella at slaughter are considered a source of carcass contamination and human infection. To assess this potential risk, the proportion of Salmonella shedders that arrive for slaughter was evaluated in a population of 1068 pigs from 24 farms. Shedding was present in 27.3% of the pigs, and the monophasic variant of Salmonella Typhimurium, an emerging zoonotic serotype, was the most prevalent (46.9%). Antimicrobial resistance (AMR) in Salmonella isolates was common, but few isolates showed AMR to antimicrobials of critical importance for humans such as third-generation cephalosporins (5%), colistin (0%), or carbapenems (0%). However, AMR to tigecycline was moderately high (15%). The efficacy of an esterified formic acid in the lairage drinking water (3 kg formic acid/1000 L) was also assessed as a potential abattoir-based strategy to reduce Salmonella shedding. It was able to reduce the proportion of shedders (60.7% in the control group (CG) vs. 44.3% in the treatment group (TG); p < 0.01). After considering clustering and confounding factors, the odds of shedding Salmonella in the CG were 2.75 (95% CI = 1.80−4.21) times higher than those of the TG, suggesting a potential efficacy of reduction in shedding as high as 63.6%. This strategy may contribute to mitigating the burden of abattoir environmental contamination.

Effects of Dietary Supplementation with Protected Sodium Butyrate on Gut Microbiota in Growing-Finishing Pigs.

The study assessed changes in the gut microbiota of pigs after dietary supplementation with protected sodium butyrate (PSB) during the growing-fattening period (≈90 days). One gram of colon content from 18 pigs (9 from the treatment group -TG- and 9 from the control group -CG-) was collected. Bacterial DNA was extracted and 16S rRNA high-throughput amplicon sequencing used to assess microbiota changes between groups. The groups shared 75.4% of the 4697 operational taxonomic units identified. No differences in alpha diversity were found, but significant differences for some specific taxa were detected between groups. The low-represented phylum Deinococcus-Thermus, which is associated with the production of carotenoids with antioxidant, anti-apoptotic, and anti-inflammatory properties, was increased in the TG (p = 0.032). Prevotellaceae, Lachnospiraceae, Peptostreptococcaceae, Peptococcaceae, and Terrisporobacter were increased in the TG. Members of these families have the ability to ferment complex dietary polysaccharides and produce larger amounts of short chain fatty acids. Regarding species, only Clostridium butyricum was increased in the TG (p = 0.048). Clostridium butyricum is well-known as probiotic in humans, but it has also been associated with overall positive gut effects (increased villus height, improved body weight, reduction of diarrhea, etc.) in weanling pigs. Although the use of PSB did not modify the overall richness of microbiota composition of these slaughter pigs, it may have increased specific taxa associated with better gut health parameters.

Effect of Edema Disease Vaccination on Mortality and Growth Parameters in Nursery Pigs in a Shiga Toxin 2e Positive Commercial Farm.

Diseases caused by Escherichia coli are recognized as major problems in the swine industry, one of them being edema disease (ED). Importantly, the current decrease in antibiotic use may cause difficulties in controlling the disorders caused by E. coli. Therefore, this study assessed the efficacy of a commercial vaccine against ED in nursery pigs from a farm with previous history of ED. A total of 1344 pigs were monitored; half of them were randomly assigned to a vaccinated group (VG) and the other half to a non-vaccinated group (NVG). The vaccine was administered at 7 days of age. Animals received a pre-starter feed with 2500 ppm of zinc oxide (ZnO) for 2 weeks and a starter feed without ZnO for another 3 weeks. Pen-group weights were recorded at 28 (weaning), 42 (end of pre-starter phase), and 63 days of life (end of nursery phase). Death/culling rates, average daily gain (ADG), and average daily feed intake (ADFI) were calculated for each group at each phase. The overall relative risk of dying/being culled for a pig in the NVG was 5 times higher than that of the VG group but increased to 12 times higher during the starter period. ADG and ADFI were also significantly higher in the VG group for that period. Vaccination against ED significantly reduced pig losses and improved ADG and ADFI, particularly when ZnO was not used.

Incidence and characterization of Clostridium difficile in a secondary care hospital in Spain

Introduction: Clostridium difficile (C. difficile) is a major nosocomial infectious agent in hospitals. Previous studies have addressed the high proportion of infection episodes that are overlooked in health care facilities. Objective: the main aim of this study was to characterize C. difficile clinical cases that occurred in a secondary care hospital during a five-month period. Material and methods: for this purpose, a total of 137 stool samples from the same number of patients with diarrhea were analyzed for the presence of C. difficile by culture techniques. An enzyme immunoassay (EIA) test for the detection of C. difficile and its toxins was also used in 50 cases (36.5%) for diagnostic purposes. Results: a total of 14 (10.2%) C. difficile isolates were obtained, of which nine (64.3%) were toxigenic. A mean incidence of 3.2 episodes of C. difficile infections (CDI) per 10,000 patients-days was estimated for the study period. Around 56% of the CDI cases were determined as hospital-acquired, whereas 44% originated in the community. Among these, only two episodes (22.2%) were detected in the hospital by the EIA test, which indicated that the hospital CDI detection protocol needed to be revised. One unusual C. difficile isolate was negative for all toxin genes examined and also for the non-toxigenic strain assay, which highlights the need to perform genome sequencing to study its pathogenicity locus insertion site organization. A stable metronidazole-resistant C. difficile strain and three strains showing multidrug resistance were detected in this study, suggesting that C. difficile antimicrobial susceptibility surveillance programs should be established in this health-care facility

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Incidence and characterization of Clostridium difficile in a secondary care hospital in Spain.

INTRODUCTION: Clostridium difficile (C. difficile) is a major nosocomial infectious agent in hospitals. Previous studies have addressed the high proportion of infection episodes that are overlooked in health care facilities. OBJECTIVE: the main aim of this study was to characterize C. difficile clinical cases that occurred in a secondary care hospital during a five-month period. MATERIAL AND METHODS: for this purpose, a total of 137 stool samples from the same number of patients with diarrhea were analyzed for the presence of C. difficile by culture techniques. An enzyme immunoassay (EIA) test for the detection of C. difficile and its toxins was also used in 50 cases (36.5%) for diagnostic purposes. RESULTS: a total of 14 (10.2%) C. difficile isolates were obtained, of which nine (64.3%) were toxigenic. A mean incidence of 3.2 episodes of C. difficile infections (CDI) per 10,000 patients-days was estimated for the study period. Around 56% of the CDI cases were determined as hospital-acquired, whereas 44% originated in the community. Among these, only two episodes (22.2%) were detected in the hospital by the EIA test, which indicated that the hospital CDI detection protocol needed to be revised. One unusual C. difficile isolate was negative for all toxin genes examined and also for the non-toxigenic strain assay, which highlights the need to perform genome sequencing to study its pathogenicity locus insertion site organization. A stable metronidazole-resistant C. difficile strain and three strains showing multidrug resistance were detected in this study, suggesting that C. difficile antimicrobial susceptibility surveillance programs should be established in this health-care facility.

Preliminary studies on isolates of Clostridium difficile from dogs and exotic pets.

BACKGROUND: Clostridium difficile infection (CDI) is recognised as an emerging disease in both humans and some animal species. During the past few years, insights into human CDI epidemiology changed and C. difficile is also considered as an emerging community-acquired pathogen. Certain ribotypes (RT) are possibly associated with zoonotic transmission. The objective of this study was to assess the presence of C. difficile in a population of pets and to characterise the isolates. RESULTS: Faecal samples from a total of 90 diarrhoeic dogs and 24 from exotic animal species (both diarrhoeic and non-diarrhoeic) were analysed. Clostridium difficile was isolated from 6 (6.7%) dogs and one reptile sample (4.2%). Four (66.7%) of the six dog strains were capable of producing toxins. Four known different RTs were detected in dogs (010, 014, 123 and 358) and a new one was found in a faecal sample of an exotic animal. This new RT isolate was negative for all toxin genes tested and belonged to sequence type 347 which has been proposed as a Clade-III member. Importantly, two dog strains showed a stable resistance to metronidazole (initial MIC values: 128 and 48 µg/ml). CONCLUSIONS: The results obtained in this study suggest the implementation of antimicrobial susceptibility surveillance programs to assess the prevalence of metronidazole resistance in dogs; molecular studies to elucidate C. difficile metronidazole resistance mechanisms are warranted. Based on the similarity between the ribotypes observed in dogs and those described in humans, the zoonotic transmission should be further explored. Furthermore, exotic animals have shown to harbor uncommon C. difficile strains which require further genomic studies.

Simultaneous infections by different Salmonella strains in mesenteric lymph nodes of finishing pigs.

BACKGROUND: Salmonellosis is a major worldwide zoonosis, and Salmonella-infected finishing pigs are considered one of the major sources of human infections in developed countries. Baseline studies on salmonellosis prevalence in fattening pigs in Europe are based on direct pathogen isolation from mesenteric lymph nodes (MLN). This procedure is considered the most reliable for diagnosing salmonellosis in apparently healthy pigs. The presence of simultaneous infections by different Salmonella strains in the same animal has never been reported and could have important epidemiological implications. RESULTS: Fourteen finishing pigs belonging to 14 farms that showed high salmonellosis prevalence and a variety of circulating Salmonella strains, were found infected by Salmonella spp, and 7 of them were simultaneously infected with strains of 2 or 3 different serotypes. Typhimurium isolates showing resistance to several antimicrobials and carrying mobile integrons were the most frequently identified in the colonized MLN. Four animals were found infected by Salmonella spp. of a single serotype (Rissen or Derby) but showing 2 or 3 different antimicrobial resistance profiles, without evidence of mobile genetic element exchange in vivo. CONCLUSION: This is the first report clearly demonstrating that pigs naturally infected by Salmonella may harbour different Salmonella strains simultaneously. This may have implications in the interpretation of results from baseline studies, and also help to better understand human salmonellosis outbreaks and the horizontal transmission of antimicrobial resistance genes.