Alda F. A. Pires

Longitudinal study of Salmonella shedding in naturally infected finishing pigs.

A 3-year longitudinal study was conducted on a multi-site farrow-to-finish production system. For each of 18 cohorts at three finishing sites, 50 pigs were randomly selected. Faecal samples were collected every 2 weeks for 16 weeks. Salmonella was cultured from 453 (6·6%) of 6836 faecal samples. The pig-level incidence of Salmonella was 20·8% (187/899 pigs). Salmonella prevalence varied between cohorts and within pigs. The adjusted Salmonella prevalence decreased over the finishing period from 6·4% to 0·8%. Intermittent detection of Salmonella was found in more than 50% of pigs that were positive at more than one collection. The finding that the majority of pigs shed intermittently has implications for surveillance and research study design when determining Salmonella status. The variability in shedding over time, as well as between and within cohorts and pigs suggests that there may be time-variant risk factors for Salmonella shedding in swine.

Longitudinal study to evaluate the association between thermal environment and Salmonella shedding in a midwestern US swine farm

The objective of this study was to document the association between the thermal environment in the barn and Salmonella shedding in finishing pigs. For this purpose, individual fecal samples from 900 finishing pigs (8 collections per pig) were repeatedly collected from 18 cohorts (50 pigs per cohort) on 3 sites of a multi-site farrow-to-finish production system in a longitudinal study. Pen temperature and humidity were measured every 2 min during the study period. The thermal parameters of interest were: hourly average temperature, minimum and maximum temperature, hourly temperature variation, temperature humidity index (THI) and cumulative number of hours/degree above and below the thermal neutral zone at the pen level prior to fecal sampling for 6 time periods (12h, 24h, 48 h, 72 h, 1 week and 1 month). Additional potential risk factors at the individual (e.g., sex, health events), cohort (e.g., mortality, morbidity, Salmonella status of the nursery) and pen level (e.g., type of pen) were also evaluated. Multilevel logistic models using generalized linear models, with random intercepts at pig, pen and cohort levels to account for clustering (individual samples nested within pigs, pigs nested within pens, pens within cohorts) were constructed. Site (A, B, C) was considered as a fixed effect in order to control for clustering within site. The outcome variable was Salmonella fecal status of the individual sample. Cold exposure (temperatures below the thermal neutral zone) and exposure to a THI>72 were both positively associated with risk Salmonella shedding. Nursery Salmonella status was positively associated with Salmonella shedding and pig age was negatively associated with Salmonella shedding. In the multilevel intercept-only model the largest proportion of model variance was associated with the individual fecal sample (44.8%) followed by cohort (24.5%), pen (20.5%) and pig (10.2%). The present study allowed the investigation of the association of time-variant thermal factors and Salmonella shedding. Interventions that target the thermal environment may have an effect on reducing Salmonella shedding in swine and also improve pig well-being and production efficiency. Alternatively, thermal parameters may be used to identify groups of pigs at high risk for Salmonella shedding. Future studies should be performed to investigate the cost-efficacy of interventions to improve the thermal environment of swine.

Assessment of a fluorescent antibody test for the detection of antibodies against epizootic bovine abortion.

The current study was directed at developing and validating an indirect fluorescent antibody test (IFAT) capable of detecting antibodies specific for the agent of epizootic bovine abortion (aoEBA). Sensitivity and specificity was determined by comparing antibody titers from 114 fetuses infected with aoEBA with 68 fetuses diagnosed with alternate infectious etiologies. Data established specificity at 100% and sensitivity at 94.7% when cutoff criteria for a positive test were assigned at a titer of ≥1,000. Potential cross-reactivity was noted in samples from 3 fetuses with antibody titers of 10 or100; all were infected with Gram-positive organisms. The remaining 65 fetuses infected with microbes other than aoEBA, and an additional 12 negative reference sera, did not have detectable titers. The IFAT-based serology assay is rapid, reproducible, and unaffected by fluid color or opacity. Total fetal immunoglobulin (Ig)G was also evaluated as an aid for diagnosing EBA. Significantly higher concentrations of IgG were identified in fetuses infected with aoEBA as compared to those with alternate infectious etiologies. The presence of IgG is a sensitive indicator of EBA and increases the specificity of FAT-based serologic diagnosis when titers are 10 or 100. Taken together, serology and IgG analyses suggest that the incidence of EBA may be underestimated.

Enumeration of salmonella in feces of naturally infected pigs

Quantification of Salmonella in asymptomatic pigs can be used to institute control measures and to assess risk of carcass contamination during slaughter. The objective of this study was to quantify the fecal concentration of Salmonella in naturally infected pigs. Individual fecal samples (positive [n=443], negative [n=1225] determined by microbiological culture) were submitted for direct quantitative real-time polymerase chain reaction (q-PCR). Direct q-PCR categorized 99.6% (1220/1225) of culture negative samples as negative. For culture positive samples, 15.4% (68/443) were detected by q-PCR, but only 3.4% (15/443) were within the direct q-PCR quantifiable range (≥ 10(3) colony-forming units [CFU]/g of feces). Of these latter samples, the concentration range was 1.06 × 10(3) to 1.73 × 10(6) CFU/g feces. Of the 15 samples with high Salmonella concentrations, seven were collected from one pig and three samples were collected from its penmates. Direct q-PCR may be an alternative to traditional culture-dependent methods for detection of pigs with high fecal concentrations of Salmonella, but not for detection of pigs shedding low concentrations of Salmonella, which represented the majority of pigs in this study. When high shedding was detected it was clustered within a single pig and its penmates. These data contribute to quantitative risk assessments of the association between concentrations of Salmonella shed by pigs during the finishing phase and risk of carcass contamination at slaughter.

Persistence of Escherichia coli in the soil of an organic mixed crop-livestock farm that integrates sheep grazing within vegetable fields

Mixed crop‐livestock farms (MCLF) integrate livestock and crops using their animals to graze crop residues and/or cover crops. MCLF are considered sustainable because grazing and the manure deposited by livestock enhance soil fertility and recycles farm nutrients. However, livestock manure may introduce enteric foodborne pathogens to the soil, which could contaminate fresh produce. Organic farmers in the United States follow the USDA National Organic Program (NOP) standards, which require 90 or 120 days between incorporating raw manure into the soil and harvest. Although not specifically addressed in NOP, organic farmers using grazing within production fields may also use this standard. The objectives of this study were to generate preharvest data to assess the die‐off of generic Escherichia coli (E. coli) in the soil, after cover crops were grazed by sheep; and assess the genetic relatedness of generic E. coli isolates between soil and sheep faecal samples. We conducted a repeated observational study to evaluate the persistence of generic E. coli, as an indicator of faecal contamination and surrogate for STEC, in the soil of two fields (A and B) on an organic MCLF. Results showed a 3.70 log10 reduction in mean generic E. coli concentration MPN in the soil of field A from the highest of 3.70 log10 MPN/g on 48 day postsheep grazing (DPS) to −0.70 log10 MPN/g on 139 DPS. Field B showed a 3.51 log10 reduction in mean generic E. coli concentration in the soil from the highest mean of 3.51 log10 MPN/g on 14 DPS to the lowest mean −0.35 log10 MPN/g on 112 DPS. STEC prevalence in the sheep flock was 4.17% (1/24). Closely related generic E. coli strains were found between soil and faecal samples. Developing research‐based waiting periods between grazing and harvest is important to inform best practices for farmers and food safety regulators.

Antimicrobial resistance trends in fecal Salmonella isolates from northern California dairy cattle admitted to a veterinary teaching hospital, 2002-2016

Nontyphoidal Salmonella infections contribute to approximately 1.2 million annual illnesses in the United States. Historical and recent outbreaks have been associated with dairy products, ground beef, and direct contact with cattle. Salmonella antimicrobial resistance (AMR) is a serious concern that can reduce successful treatment of infections, increasing recovery time, medical costs, and mortality rates in humans and animals. This highlights the need to track AMR in Salmonella isolated from cattle to improve treatment plans, manage trends in AMR, and prevent future AMR development. A total of 242 Salmonella isolates were retrieved from 9,162 cattle fecal samples submitted to the University of California, Davis Veterinary Medical Teaching Hospital from 2002 to 2016. These isolates were tested for antimicrobial susceptibility using a standardized broth dilution panel. Multidrug resistance (MDR) to three or more classes of antimicrobials was observed in 50.8% of isolates, and the most common MDR pattern was amoxicillin-ampicillin-cefoxitin-ceftiofur-ceftriaxone-chloramphenicol-streptomycin-tetracycline (23.2%). There were significantly greater odds for antimicrobial resistance to aminoglycosides (OR: 2.03, 95% CI: 1.1–3.7), penicillins (OR: 1.87, 95% CI: 1.007–3.5), and tetracyclines (OR: 1.87, 95% CI: 1.017–3.4) for the 2002–2009 period when compared to the 2010–2016 period. The most prevalent MDR serotypes were Newport (100% MDR, n = 52), Typhimurium (100%, n = 20), and Dublin (71% MDR, n = 46). Risk factors associated with higher odds for isolating MDR Salmonella included isolates from calves when compared to adult cattle (OR: 22.0; 95% C.I.: 3.9–125.7), and isolates obtained from cattle suspect of having salmonellosis versus from the infectious disease control surveillance program (OR:13.7; 95%C.I.: 2.8–66.8). Despite a temporal trend for reduced AMR to most antimicrobial drug classes, a lack of this observed in the 2002–2009 period when compared to the 2010–2016 period for important drug classes such as cephalosporins (OR: 1.6, 95% CI: 0.87–3.1), and a trend for temporal increase in resistant to quinolones drugs (P value 0.004) highlight the relevance of AMR surveillance in cattle with Salmonella infections with the aim of targeting future prophylactic interventions.

Phenotypic and Genotypic Diversity of Salmonella in Finishing Swine.

Salmonella enterica (nontyphoidal) is one of the major causes of foodborne diseases in the United States and worldwide. Molecular typing methods are significant tools used to better understand the transmission and ecology of Salmonella in order to implement pre-harvest control measures. The objectives of this study were to describe the Salmonella genotypes, the distribution of isolate subtypes from different ecological niches (i.e., barn environment, nursery, and individual pigs) and their evolution over time in a longitudinal study conducted in three finishing sites (housing pigs from 10 weeks of age until slaughter at 24-26 weeks of age). Among the 107 Salmonella isolates submitted for pulsed-field gel electrophoresis (PFGE) analysis, there were 25 distinct subtypes. PFGE genotyping results were consistent with the serotype findings. A large number of distinguishable PFGE patterns (i.e., within the same serovar) were observed and different combinations of subtypes were identified within and across sites and cohorts. New subtypes may result of the introduction of new strains, genetic changes, or ongoing transmission of evolved strains within the production system. The same subtypes were detected intermittently during the study period, which suggests the persistence of indistinguishable subtypes in this production system. In addition, this study suggests persistence of the same subtype over several cohorts of pigs and potential residual contamination from the barn. Factors affecting adaptation and transmission of Salmonella within and among ecological systems (e.g., finishing pigs, nursery, and environment) should be further investigated. Understanding genotypic diversity of Salmonella in different ecological niches during pre-harvest may contribute to the development of more targeted and cost effective control programs during nursery and finishing phases.