Factors Associated with Salmonella Prevalence in U.S. Swine Grower-Finisher Operations, 2012.

Nontyphoidal Salmonella is an important foodborne pathogen with diverse serotypes occurring in animal and human populations. The prevalence of the organism on swine farms has been associated with numerous risk factors, and although there are strong veterinary public health controls for preventing Salmonella from entering food, there remains interest in eradicating or controlling the organism in the preharvest environment. In this study, using data collected via the U.S. Department of Agriculture (USDA) National Animal Health Monitoring System Swine 2012 study, we describe nontyphoidal Salmonella and specific serotype prevalence on U.S. grower-finisher swine operations and investigate associations between Salmonella detection and numerous factors via multiple correspondence analysis (MCA) and regression analysis. MCA plots, complementary to univariate analyses, display relationships between covariates and Salmonella detection at the farm level. In the univariate analysis, Salmonella detection varied with feed characteristics and farm management practices, reports of diseases on farms and vaccinations administered, and administration of certain antimicrobials. Results from the univariate analysis reinforce the importance of biosecurity in managing diseases and pathogens such as Salmonella on farms. All multivariable regression models for the likelihood of Salmonella detection were strongly affected by multicollinearity among variables, and only one variable, pelleted feed preparation, remained in the final model. The study was limited by its cross-sectional nature, timelines of data collection, and reliance on operator-reported data via a convenience sample.

Prevalence of Salmonella enterica, Listeria monocytogenes, and pathogenic Escherichia coli in bulk tank milk and milk filters from US dairy operations in the National Animal Health Monitoring System Dairy 2014 study.

The dairy farm environment is a well-documented reservoir for zoonotic pathogens such as Salmonella enterica, Shiga-toxigenic Escherichia coli, and Listeria monocytogenes, and humans may be exposed to these pathogens via consumption of unpasteurized milk and dairy products. As part of the National Animal Health Monitoring System Dairy 2014 study, bulk tank milk (BTM, n = 234) and milk filters (n = 254) were collected from a total of 234 dairy operations in 17 major dairy states and analyzed for the presence of these pathogens. The invA gene was detected in samples from 18.5% of operations and Salmonella enterica was isolated from 18.0% of operations. Salmonella Dublin was detected in 0.7% of operations. Sixteen Salmonella serotypes were isolated, and the most common serotypes were Cerro, Montevideo, and Newport. Representative Salmonella isolates (n = 137) were tested against a panel of 14 antimicrobials. Most (85%) were pansusceptible; the remaining were resistant to 1 to 9 antimicrobials, and within the resistant strains the most common profile was resistance to ampicillin/clavulanic acid, ampicillin, cefoxitin, ceftiofur, ceftriaxone, chloramphenicol, streptomycin, sulfisoxazole, and tetracycline. Listeria spp. were isolated from 19.9% of operations, and L. monocytogenes was isolated from 3.0% of operations. Serogroups 1/2a and 1/2b were the most common, followed by 4b and 4a. One or more E. coli virulence genes were detected in the BTM from 30.5% of operations and in the filters from 75.3% of operations. A combination of stx2, eaeA, and γ-tir genes was detected in the BTM from 0.5% of operations and in the filters from 6.6% of operations. The results of this study indicate an appreciable prevalence of bacterial pathogens in BTM and filters, including serovars known to infect humans.

Prevalence, antimicrobial resistance, and molecular characterization of Campylobacter spp. in bulk tank milk and milk filters from US dairies.

Campylobacter spp. are frequently isolated from dairy cows as commensal organisms. Sporadic Campylobacter infections in humans in the United States are generally attributed to poultry, but outbreaks are also commonly associated with dairy products, particularly unpasteurized or raw milk. Bulk tank milk samples and milk filters from US dairy operations were collected during the National Animal Health Monitoring System Dairy 2014 study and analyzed using real-time PCR and traditional culture techniques for the presence of thermophilic Campylobacter species. The weighted prevalence of operations from which we detected Campylobacter spp. in either bulk tank milk or milk filters was 24.9%. We detected Campylobacter spp. in a higher percentage of operations with 100-499 cows (42.8%) and 500 or more cows (47.5%) than in operations with 30-99 cows (6.5%). Campylobacter spp. were also more frequently detected in operations in the west than the east (45.9 and 22.6%, respectively). We isolated Campylobacter spp. from approximately half of PCR-positive samples, representing 12.5% (weighted prevalence) of operations. The majority (91.8%) of isolates were C. jejuni, but C. lari and C. coli were also isolated. We detected resistance to tetracycline in 68.4% of C. jejuni isolates, and resistance to ciprofloxacin and nalidixic acid in 13.2% of C. jejuni isolates. Based on pulsed-field gel electrophoresis, we found that dairy-associated C. jejuni were genotypically diverse, although clonal strains were isolated from different geographic regions. These results suggest that bulk tank milk can be contaminated with pathogenic Campylobacter spp., and that the consumption of unpasteurized or raw milk presents a potential human health risk.

Prevalence and Antimicrobial Resistance of Salmonella Isolated from Cattle Feces in United States Feedlots in 2011.

The objective of this study was to determine the prevalence and characteristics of Salmonella spp. isolated from feces of cattle in feedlots in the United States. Fecal samples were collected from up to three pens of cattle in each of 68 feedlots in 12 states. Samples included up to 25 individual fecal pats from the pen floors and up to five composite samples from the floors of the same pens. The prevalence of Salmonella-positive samples was 9.1% (460/5050) and 11.3% (114/1009) for individual and composite samples, respectively. The prevalences of Salmonella at the pen level were 35.6% (72/202) and 22.8% (46/202) for individual and composite samples, respectively. Dietary factors, including inclusion of cottonseed hulls, coccidiostats, and antimicrobial drugs, were associated with differences in prevalence of Salmonella isolation. Overall, 32 serotypes of Salmonella were identified, but six serotypes accounted for 69.1% (495/716) of the isolates. Nearly two-thirds (64.7%, 44/68) of feedlots had at least one positive sample. All isolates were evaluated for susceptibility to a panel of 15 antimicrobial drugs. Most isolates (74.4%, 533/716) were susceptible to all antimicrobial drugs in the panel. When resistance was detected, it was most commonly to tetracycline (21.7%, 155/716 of isolates) or sulfisoxazole (12.4%, 89/716 of isolates). Less than 10% of the isolates were resistant to any other antimicrobials in the panel. The results of this study indicate that the prevalence of Salmonella in individual fecal samples was less than 10%, but that Salmonella is widely distributed among feedlot cattle. Furthermore, when Salmonella is present in feedlot cattle, there is a low occurrence of antimicrobial resistance with the exception of tetracycline and sulfisoxazole. More research is indicated to understand the ecology of Salmonella and antimicrobial resistance, when present, in cattle-feeding operations.

Salmonella Prevalence and Antimicrobial Susceptibility from the National Animal Health Monitoring System Sheep 2011 Study.

Salmonella is a major cause of foodborne illness and can cause clinical disease in animals. Understanding the on-farm ecology of Salmonella will be helpful in decreasing the risk of foodborne transmission. An objective of this study was to determine the prevalence of Salmonella among fecal samples collected on sheep operations in the United States. Another objective was to compare the use of composite fecal samples with fecal samples collected from individual sheep as a tool for screening sheep flocks for Salmonella. Sheep fecal samples (individual and composite) were collected on operations in 22 states. Salmonella isolates were characterized with regard to species, serotype, and antimicrobial susceptibility profile. Most operations (72.1%) had at least one positive sample and overall 26.9% of samples were positive. The percentage of positive samples varied by animal age class. Composite and individual samples gave similar results. The majority of the isolates (94%) were Salmonella enterica subspecies diarizonae serotype 61:-:1,5,7. Nearly all of the isolates (91.2%) tested for antimicrobial susceptibility were susceptible to all antimicrobials in the panel. The findings suggest that salmonellae typically associated with foodborne disease transmission are infrequently found on sheep operations in the United States.

Prevalence of internal parasites in beef cows in the United States: Results of the National Animal Health Monitoring System's (NAHMS) beef study, 2007-2008.

During the United States Department of Agriculture (USDA) National Animal Health Monitoring System's (NAHMS) 2007-2008 beef study, 567 producers from 24 US States were offered the opportunity to collect fecal samples from weaned beef calves and have them evaluated for the presence of parasite eggs (Phase 1). Participating producers were provided with instructions and materials for sample collection. Up to 20 fresh fecal samples were collected from each of the 99 participating operations. Fresh fecal samples were submitted to one of 3 randomly assigned laboratories for evaluation. Upon arrival at the laboratories, all samples were processed for the enumeration of strongyle, Nematodirus, and Trichuris eggs using the modified Wisconsin technique. The presence or absence of coccidian oocysts and tapeworm eggs was also noted. In submissions where the strongyle eggs per gram exceeded 30, aliquots from 2 to 6 animals were pooled for DNA extraction. Extracted DNA was subjected to genus level polymerase chain reaction (PCR) identification for the presence of Ostertagia, Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. In this study, 85.6% of the samples had strongyle type, Nematodirus, and Trichuris eggs. Among the samples evaluated, 91% had Cooperia, 79% Ostertagia, 53% Haemonchus, 38% Oesophagostomum, 18% Nematodirus, 7% Trichuris, and 3% Trichostrongylus. The prevalence of coccidia and tapeworm eggs was 59.9% and 13.7%, respectively.

Pendant l'étude de 2007­2008 chez les bovins effectuée par le Système national de surveillance des maladies animales (NAHMS) du Département de l'agriculture des États-Unis (USDA), 567 producteurs provenant de 24 états américains se sont vus offrir l'opportunité de prélever des échantillons de fèces de veaux sevrés et de les faire analyser pour la présence d'oeufs de parasite (Phase 1). On a fourni aux producteurs participants les instructions et le matériel pour le prélèvement d'échantillon. Jusqu'à 20 échantillons de fèces fraiches furent prélevés de chacune des 99 opérations participantes. Les échantillons de fèces fraiches furent soumis de manière aléatoire pour évaluation à l'un des trois laboratoires participants. Suite à l'arrivée au laboratoire, tous les échantillons étaient traités pour énumération des strongles, de Nematodirus, et d'oeufs de Trichuris en utilisant la technique de Wisconsin modifiée. La présence ou l'absence d'ookystes de coccidie et d'oeufs de vers plats furent également notées. Dans les échantillons soumis et dont le nombre d'oeufs de strongles par gramme dépassait 30, des aliquots de 2 à 6 animaux étaient regroupés pour extraction de l'ADN. L'ADN extrait était soumis à une réaction d'amplification en chaine par la polymérase (PCR) pour une identification au genre de la présence d'Ostertagia, de Cooperia, d'Haemonchus, d'Oesophagostomum, et de Trichostrongylus. Dans la présente étude, 85,6 % des échantillons avaient des strongles, du Nematodirus, et des oeufs de Trichuris. Parmi les échantillons évalués, 91 % avaient du Cooperia, 79 % de l'Ostertagia, 53 % de l'Haemoncus, 38 % de l'Oesophagostomum, 18 % du Nematodirus, 7 % du Trichuris, et 3 % du Trichostrongylus. Les prévalences de coccidies et d'oeufs de vers plats étaient respectivement de 59,9 % et 13,7 %.(Traduit par Docteur Serge Messier).

Effectiveness of current anthelmintic treatment programs on reducing fecal egg counts in United States cow-calf operations.

During the United States Department of Agriculture (USDA) National Animal Health Monitoring System's (NAHMS) 2007-2008 beef study, producers from 24 states were offered the opportunity to evaluate their animals for internal parasites and for overall responses to treatment with anthelmintics. A lapse of 45 d was required between initial sampling and any previous treatments. Choice of anthelmintic (oral benzimidazoles, and both injectable and pour-on endectocides) was at the discretion of the producer so as not to alter the local control programs. Fresh fecal samples were collected from 20 animals, or from the entire group if less than 20, then randomly assigned to 1 of 3 participating laboratories for examination. Analyses consisted of double centrifugation flotation followed by enumeration of strongyle, Nematodirus, and Trichuris eggs (the presence of coccidian oocysts and tapeworm eggs was also noted). Where strongyle eggs per gram (epg) exceeded 30, aliquots from 2 to 6 animals were pooled for egg isolation and polymerase chain reaction (PCR) analysis for the presence of Ostertagia, Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. Results from 72 producers (19 States) indicated that fecal egg count reductions were < 90% in 1/3 of the operations. All operations exhibiting less than a 90% reduction had used pour-on macrocyclic lactones as the anthelmintic treatment. While some of these less than expected reductions could have been the result of improper drug application, PCR analyses of the parasite populations surviving treatment, coupled with follow-up studies at a limited number of sites, indicated that less than expected reductions were most likely due to anthelmintic resistance in Cooperia spp. and possibly Haemonchus spp.

Pendant l'étude de 2007­2008 chez les bovins effectuée par le Système national de surveillance des maladies animales (NAHMS) du Département de l'agriculture des États-Unis (USDA), des producteurs provenant de 24 états américains se sont vus offrir l'opportunité de faire évaluer leurs animaux pour la présence de parasites internes et pour leur réponse globale à un traitement avec des anthelminthiques. Un délai de 45 j était requis entre l'échantillonnage initial et un traitement antérieur. Le choix d'un anthelminthique (benzimidazole oral, et des endectocides injectables et en solution à verser) était à la discrétion du producteur afin de ne pas altérer les programmes de contrôle locaux. Des échantillons de fèces fraiches ont été prélevés de 20 animaux, ou de tout le groupe si moins de 20, puis ils ont été acheminés de manière aléatoire à un des trois laboratoires participants pour fin d'examen. L'analyse consistait en une double centrifugation par flottaison suivie d'une énumération des strongles, de Nematodirus, et d'oeufs de Trichuris (la présence d'ookystes de coccidie et d'oeufs de vers plats fut également notée). Lorsque le nombre d'oeufs de strongles par gramme dépassait 30, des aliquots de 2 à 6 animaux étaient regroupés pour isolement des oeufs et et soumis à une réaction d'amplification en chaine par la polymérase (PCR) pour détecter la présence d'Ostertagia, de Cooperia, d'Haemonchus, d'Oesophagostomum, et de Trichostrongylus. Les résultats provenant de 72 producteurs (19 états) indiquent que les réductions dans le dénombrement des oeufs dans les fèces étaient de < 90 % dans le tiers des opérations. Toutes les opérations montrant une réduction de moins de 90 % avaient utilisé des lactones macrocycliques en solution à verser comme traitement anthelminthique. Alors que certaines de ces réductions moindres que prévues puissent être le résultat d'une mauvaise application du produit, les analyses par PCR des populations de parasites survivantes au traitement, combinées aux études de suivis à un nombre limité de sites, indiquent que les réductions moindres que prévues étaient fort probablement dues à la résistance aux anthelminthiques chez Cooperia spp. et possiblement Haemonchus spp.(Traduit par Docteur Serge Messier).

Factors affecting use of veterinarians by small-scale food animal operations.

OBJECTIVE: To identify factors associated with use of a veterinarian by small-scale food animal operations. DESIGN: Cross-sectional descriptive survey. SAMPLE: 16,000 small-scale farm or ranch operations in all 50 states. PROCEDURES: Surveys were conducted via mail or telephone during 2011 for small-scale operations (gross annual agricultural sales between $10,000 and $499,999) in which an animal or animal product comprised the highest percentage of annual sales. RESULTS: 8,186 (51.2%) operations responded to the survey; 7,849 surveys met the inclusion criteria. For 6,511 (83.0%) operations, beef cattle were the primary animal species. An estimated 82.1% of operations (95% confidence interval [CI], 81.1% to 83.0%) had a veterinarian available ≤ 29 miles away; 1.4% (95% CI, 1.2% to 1.7%) did not have a veterinarian available within 100 miles of the operation. Operations for which the nearest veterinarian was ≥ 100 miles away or for which a veterinarian was not available were located in 40 US states. Overall, 61.7% of operations (95% CI, 60.6% to 62.9%) had used a veterinarian during the 12 months prior to the survey. Producers with college degrees were significantly more likely to use a veterinarian (675%) versus those who did not complete high school (52.9%). CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study indicated most small-scale operations had adequate access to veterinarians during 2011, but there seemed to be localized shortages of veterinarians in many states.

Prevalence of Escherichia coli O-types and Shiga toxin genes in fecal samples from feedlot cattle.

While efforts to control foodborne illness associated with the Shiga toxin-producing Escherichia coli (E. coli) O157 through processes and procedures implemented at harvest facilities have been very successful, there is concern about the burden of illness associated with other Shiga toxin-producing E. coli. The U.S. Department of Agriculture Food Safety and Inspection Service announced plans to classify an additional six non-O157 Shiga toxin-producing E. coli as adulterants. Little is known about the prevalence and distribution of these E. coli in the animal production environment. An investigation of the prevalence of O157 and the six major non-O157 E. coli serogroups was conducted in 21 feedlots over the period July 2011 to October 2011. Individual fecal swabs were collected from cattle approximately 60 days after their arrival in the feedlot and were pooled for evaluation using a polymerase chain reaction assay to identify the presence of seven E. coli O-types (O157, O45, O103, O121, O145, O26, and O111) and four virulence genes (stx1, stx2, eaeA, and ehxA). Overall, 1145 fecal pools were evaluated, with 506 (44.2%) being positive for one or more of the E. coli O-serogroups. The pool prevalences for E. coli O157, O45, O26, O103, O121, O145, and O111 were 19.7%, 13.8%, 9.9%, 9.3%, 5.5%, 1.1%, and 0.5%, respectively. Nearly all pools were positive for ehxA (99.7%) or stx2 (98.6%). The pool level prevalence for stx1 and eae was 65.5% and 69.3%, respectively. Pools that were positive for one or more of the other E. coli O-serogroups were 1.37 times more likely to be positive for E. coli O157. Conversely, pools that were positive for E. coli O157 were 1.43 times more likely to be positive for at least one of the other E. coli O-serogroups evaluated. These data will be useful to understand the expected prevalence of potential Shiga toxin-producing E. coli in cattle feedlots.

Farm-level associations with the shedding of Salmonella and antimicrobial-resistant Salmonella in U.S. dairy cattle.

Salmonella enterica is the leading cause of foodborne-related deaths and hospitalizations within the United States. Infections caused by antimicrobial-resistant (AMR) strains are associated with higher hospital costs and case fatality. The objective for this study was to determine the association of management practices with the recovery of Salmonella and AMR Salmonella on dairy herds. Individual adult cow fecal samples and/or composite fecal samples were collected from 265 dairy herds in 17 states. Samples were cultured for Salmonella, and the MIC was determined for 15 antimicrobials. Herds were classified as Salmonella positive if at least one isolate was recovered, and AMR Salmonella positive if at least one resistant isolate was recovered. Questionnaires regarding management practices were administered to herd operators, and a subset of practices was selected based on subject knowledge and prior research. Data on preventive and therapeutic antimicrobial usage were included in the analysis. Logistic regression models were used to determine which practices were significantly (p<0.05) associated with each herd classification. A total of 124 and 25 herds were classified as Salmonella positive and AMR Salmonella positive, respectively. Variables significantly associated with Salmonella-positive herds included using sprinklers or misters for heat abatement (OR=2.8; CI: 1.6-4.9), feeding anionic salts to cows (OR=1.9; CI: 1.1-3.5), and feeding ionophores to cows (OR=2.1; CI: 1.2-3.7). Herds that used a broadcast/solid spread had lower odds (OR=0.26; CI: 0.11-0.63) of being Salmonella positive. Herds with at least one resistant isolate were more likely to have used composted/dried manure for bedding relative to herds with only susceptible isolates (OR=3.6; CI: 1.2-11.0). These results can be useful to focus additional research aimed at decreasing the prevalence of Salmonella and AMR Salmonella on U.S. dairy herds.

Prevalence of Salmonella enterica, Listeria monocytogenes, and Escherichia coli virulence factors in bulk tank milk and in-line filters from U.S. dairies.

The zoonotic bacteria Salmonella enterica, Listeria monocytogenes, and Escherichia coli are known to infect dairy cows while not always causing clinical signs of disease. These pathogens are sometimes found in raw milk, and human disease outbreaks due to these organisms have been associated with the consumption of raw milk or raw milk products. Bulk tank milk (BTM) samples (536) and in-line milk filters (519) collected from dairy farms across the United States during the National Animal Health Monitoring System's Dairy 2007 study were analyzed by real-time PCR for the presence of S. enterica and pathogenic forms of E. coli and by culture techniques for the presence of L. monocytogenes. S. enterica was detected in samples from 28.1% of the dairy operations, primarily in milk filters. Salmonella was isolated from 36 of 75 PCR-positive BTM samples and 105 of 174 PCR-positive filter samples, and the isolates were serotyped. Cerro, Kentucky, Muenster, Anatum, and Newport were the most common serotypes. L. monocytogenes was isolated from 7.1% of the dairy operations, and the 1/2a complex was the most common serotype, followed by 1/2b and 4b (lineage 1). Shiga toxin genes were detected in enrichments from 15.2% of the BTM samples and from 51.0% of the filters by real-time PCR. In most cases, the cycle threshold values for the PCR indicated that toxigenic strains were not a major part of the enrichment populations. These data confirm those from earlier studies showing significant contamination of BTM by zoonotic bacterial pathogens and that the consumption of raw milk and raw milk products presents a health risk.

A multilaboratory evaluation of a commercial enzyme-linked immunosorbent assay test for the detection of antibodies against Mycobacterium avium subsp. paratuberculosis in cattle.

Five laboratories participated in a study to evaluate sources of variation in results from an enzyme-linked immunosorbent assay (ELISA) for antibodies against Mycobacterium avium subsp. paratuberculosis. Each laboratory repeatedly tested duplicates of a negative, positive (P), and high-positive (HP) serum sample, which were supplied by the United States Department of Agriculture: Animal and Plant Health Inspection Service: Veterinary Services, National Veterinary Services Laboratories, Ames, IA, on all 96-well microtiter plates when routinely testing other samples for M. avium subsp. paratuberculosis antibodies. These 3 sera were aliquoted and sent to the 5 participating laboratories. This study focused on variation in test results because of assay reagents and laboratory techniques and did not account for biologic variability associated with the time course of infection in cattle. Overall, results from 868 microtiter plates were used in the study. For each sample a sample-to-positive (S/P) ratio was calculated according to the manufacturer's directions. The S/ P ratio for the P sample ranged from 0.06 to 1.039 (mean = 0.466 and 0.484 for wells 1 and 2, respectively) and those for the HP sample ranged from 2.446 to 8.727 (mean = 4.027 and 3.980 for wells 1 and 2, respectively). The majority of the variation in S/P ratio for the P sample was attributed to kit lot (37.5%), followed by random (unexplained) error (27.0%), laboratory (18.3%), and kit lot by laboratory (11.9%). By eliminating plates in which the separation between negative and positive control ODs was less than 0.4, the proportion of variation attributed to laboratory was reduced markedly. These results confirm that there is variability in M. avium subsp. paratuberculosis ELISA results and that several sources contribute to the observed variability. The study gives a relative estimate of the contribution of various sources to the overall variability observed in the M. avium subsp. paratuberculosis ELISA results with kit lot being a primary contributor. Similar data for other ELISA tests for antibodies to M. avium subsp. paratuberculosis or other antigens also should be developed.