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To minimize the impacts of COVID-19 and to keep campus open, Cornell University’s Ithaca, NY, campus implemented a comprehensive process to monitor COVID-19 spread, support prevention practices, and assess early warning indicators linked to knowledge, behaviors, and attitudes of campus community members. The integrated surveillance approach informed leadership and allowed for prompt adjustments to university policies and practices through evidence-based decisions. This approach enhanced healthy behaviors and promoted the well-being and safety of all community members. (Am J Public Health. 2022;112(7):980–984. https://doi.org/10.2105/AJPH.2022.306838 )

Curbing the coronavirus disease 2019 (COVID-19) pandemic requires a thorough understanding of risk factors for transmission of SARS-CoV-2, the etiologic agent. Institutions of higher education present unique challenges for controlling disease spread because of features inherent to these settings. Our objective was to determine risk factors for SARS-CoV-2 infection among a university student population in the northeastern USA during the spring and fall 2021 semesters, using the case-control study design. Cases were defined as students with a newly diagnosed SARS-CoV-2 infection detected either through the robust PCR-based surveillance testing program on campus or through healthcare testing if symptoms compatible with COVID-19 were present. Controls were defined as students with negative SARS-CoV-2 status, based on consistently negative PCR results at the time of selection. A comprehensive questionnaire was administered to each student enrolled in the study, covering a broad range of campus life activities. A total of 446 cases and 1,185 controls were included in this study. Multivariable logistic regression analysis showed that recent party attendance (adjusted OR = 2.3, p.0001), recently visiting a bar (aOR = 1.6, p = .007), living in a campus residence hall (aOR = 1.6, p = .001), fraternity/sorority membership (aOR = 1.8, p = .002), and recent travel (aOR = 1.3, p = .04) were associated with being a COVID-19 case. Having an on-campus job was negatively associated with being a COVID-19 case (aOR = 0.6, p = .0003). Among cases, the most commonly reported symptoms were cough (43.9%), fatigue (38.1%) and sore throat (30.3%). These findings can be used to inform the development of COVID-19 mitigation strategies and public health outreach efforts in university settings, thus reducing SARS-CoV-2 transmission among students and helping to preserve the vital education and research missions of these institutions.

As dairy herd sizes become larger and the organization of the business more complex, targeting communication and education to enhance animal care becomes more difficult. The purpose of this study was to describe selected demographics of calf care employees on large (500 animals) and small (501 animals) dairy farms that raise their own calves. Two to 8 individuals per farm involved with calf care, including owners, veterinarians, and calf managers, feeders, and treaters, were interviewed in either English or Spanish. Interviews were conducted in person on 53 dairy farms located in Arizona, Idaho, New York, Oregon, and Washington State. The number of preweaned calves on the farm ranged from 9 to 1,500 (median = 93). A total of 224 individuals were interviewed across 8 job titles. As farm size increased, personnel structure became more complex. Farms with100 preweaned calves were 15 times more likely to have a calf manager title compared with farms with ≤100 preweaned calves. Eight farms designated the same person as calf manager, treater, and feeder, all with ≤100 preweaned calves. Thirty-two (60%) of the farms had at least 1 full-time calf feeder. Almost 30% of owners and over 40% of veterinarians interviewed were over 50 yr of age, whereas over 40% of the calf managers, feeders, and treaters were under 30 yr of age. Seventy-three percent of feeders and 72% of treaters spoke Spanish at home. For languages in which interviewees were comfortable speaking, more than 30% of owners and 33% of veterinarians were comfortable communicating in Spanish. For calf care employees, 60% of calf managers, 42% of feeders, and 38% of treaters were bilingual (English and Spanish), but most (72%) preferred to be interviewed in Spanish. The level of education varied by job title for those interviewed, but most of the calf care team had high school or less education. However, some diversity was observed in educational background within job title with almost 38% of the calf managers having at least some college education. The majority of feeders (88%) and treaters (83%) reported being trained by another employee and 66 and 58%, respectively, had not received any continuing education in the previous year. With the amount of diversity seen on these farms, understanding employees' educational backgrounds, language, and generational differences may be valuable when developing training for new procedures for animal health or other aspects of animal care.

The objective of this study was to evaluate the longitudinal effect of enrofloxacin or tulathromycin use in calves at high risk of bovine respiratory disease (BRD) on antimicrobial resistance genes and mutation in quinolone resistance-determining regions (QRDR) in fecal E. coli. Calves at high risk of developing BRD were randomly enrolled in one of three groups receiving: (1) enrofloxacin (ENR; n = 22); (2) tulathromycin (TUL; n = 24); or (3) no treatment (CTL; n = 21). Fecal samples were collected at enrollment and at 7, 28, and 56 days after beginning treatment, cultured for Escherichiacoli (EC) and DNA extracted. Isolates were screened for cephalosporin, quinolone and tetracycline resistance genes using PCR. QRDR screening was conducted using Sanger sequencing. The only resistance genes detected were aac(6′)Ib-cr (n = 13), bla-CTX-M (n = 51), bla-TEM (n = 117), tetA (n = 142) and tetB (n = 101). A significantly higher detection of gyrA mutated at position 248 at time points 7 (OR = 11.5; P value = 0.03) and 28 (OR = 9.0; P value = 0.05) was observed in the ENR group when compared to calves in the control group. Our findings support a better understanding of the potential impacts from the use of enrofloxacin in calves on the selection and persistence of resistance.

The increasing concerns with antimicrobial resistance highlights the need for studies evaluating the impacts of antimicrobial use in livestock on antimicrobial resistance using new sequencing technologies. Through shotgun sequencing, we investigated the changes in the fecal microbiome composition and function, with a focus on functions related to antimicrobial resistance, of dairy calves. Heifers 2 to 3 weeks old, which were not treated with antibiotics by the farm before enrollment, were randomly allocated to one of three study groups: control (no treatment), enrofloxacin, or tulathromycin. Fecal samples were collected at days 4, 14, 56 and 112 days after enrollment, and DNA extraction and sample preparation and sequencing was conducted. The effect of antibiotic treatment on each taxon and functional level by time (including Day 0 as a covariate) revealed few changes in the microbiota. At the genus level, enrofloxacin group had higher abundance of Blautia, Coprococcus and Desulfovibrio and lower abundance of Bacteroides when compared to other treatment groups. The SEED database was used for functional analyses, which showed that calves in the enrofloxacin group started with a higher abundance of “Resistance to antibiotics and toxic compounds” function on Day 0, however an increase in antibiotic resistance genes after treatment with enrofloxacin was not observed. “Resistance to Fluoroquinolones” and “Erythromycin resistance”, of relevance given the study groups, were not statistically different in abundance between treatment groups. “Resistance to fluoroquinolones” increased during the study period regardless of treatment group. Despite small differences over the first weeks between treatment groups, at Day 112 the microbiota composition and functional profile was similar among all study groups. These findings show that metaphylaxis treatment of dairy calves with either enrofloxacin or tulathromycin have minimal impacts on the microbial composition and functional microbiota of dairy calves over time.

The increasing concerns with antimicrobial resistance highlights the need for studies evaluating the impacts of antimicrobial use in livestock on antimicrobial resistance using new sequencing technologies. Through shotgun sequencing, we investigated the changes in the fecal microbiome composition and function, with a focus on functions related to antimicrobial resistance, of dairy calves. Heifers 2 to 3 weeks old, which were not treated with antibiotics by the farm before enrollment, were randomly allocated to one of three study groups: control (no treatment), a single treatment of enrofloxacin, or a single treatment of tulathromycin. Fecal samples were collected at days 4, 14, 56 and 112 days after enrollment, and DNA extraction and sequencing was conducted. The effect of antibiotic treatment on each taxon and genetic functional level by time (including Day 0 as a covariate) revealed few changes in the microbiota. At the genus level, enrofloxacin group had higher relative abundance of Blautia, Coprococcus and Desulfovibrio and lower abundance of Bacteroides when compared to other study groups. The SEED database was used for genetic functional analyses, which showed that calves in the enrofloxacin group started with a higher relative abundance of "Resistance to antibiotics and toxic compounds" function on Day 0, however an increase in antibiotic resistance genes after treatment with enrofloxacin was not observed. "Resistance to Fluoroquinolones" and "Erythromycin resistance", of relevance given the study groups, were not statistically different in relative abundance between study groups. "Resistance to fluoroquinolones" increased during the study period regardless of study group. Despite small differences over the first weeks between study groups, at Day 112 the microbiota composition and genetic functional profile was similar among all study groups. In our study, enrofloxacin or tulathromycin had minimal impacts on the microbial composition and genetic functional microbiota of calves over the study period.

Understanding farmers' behavior, motivations, and perceptions toward antimicrobial use can influence how veterinarians translate research into practice and guide effective ways of implementing protocols. A multidisciplinary team investigated behavioral tendencies of New York dairy farmers toward antimicrobial use by administering a survey modeled with the reasoned action approach. This approach is a framework from social psychology containing the constructs attitude, perceived norms, and perceived behavioral control, and is used in combination with structural equation modeling to determine what drives intentions. Multiple indicators and multiple causes (MIMIC) models were then used to determine the effects of beliefs on their underlying constructs. The objective of the study was to provide direct and indirect measures of the constructs using survey data to determine importance of and associations with intention to use antimicrobials prudently. The structural equation model indicated that perceived behavioral control explained intention. Thus, farmers who feel capable of prudent use expressed positive intentions. Attitude and perception of others also had influence to a lesser extent. MIMIC models showed that the most important attributes of instrumental attitude were increasing profitability, decreasing risk of residues, and increasing herd health. Contributing attributes of affective attitude were job satisfaction, decreasing resistance, and increasing milk production. For perceived norms, the attributes were opinions/approval of family and peers, veterinarians, and milk processors. Finally, for perceived behavioral control, attributes focused on saving money on labor and treatment, ability to fit into the daily routine, and effectiveness with veterinary guidance. In conclusion, the best approach for adoption of practices might be presentation of examples of successful strategies by other producers, particularly in peer groups. In addition, veterinarians should provide the tools and guidance needed to produce economic gain, reduction of risks associated with residues and resistance, and positive experiences when using the tactics.

Feeding drug residue-containing milk to calves is common worldwide and no information is currently available on the impact on the functional profile of the fecal microbiota. Our objective was to characterize the functional profile of the fecal microbiota of preweaned dairy calves fed raw milk with residual concentrations of antimicrobials commonly found in waste milk from birth to weaning. Calves were assigned to a controlled feeding trial being fed milk with no drug residues or milk with antibiotic residues. Fecal samples collected from each calf once a week starting at birth, prior to the first feeding in the trial, until 6 weeks of age. Antibiotic residues resulted in a significant difference in relative abundance of microbial cell functions, especially with genes linked with stress response, regulation and cell signaling, and nitrogen metabolism. These changes could directly impacts selection and dissemination of virulence and antimicrobial. Our data also identified a strong association between age in weeks and abundance of Resistance to Antibiotics and Toxic Compounds. Findings from this study support the hypothesis that drug residues, even at very low concentrations, impact the gut microbiota of calves and result in changes in the functional profile of microbial populations.

SUMMARYAlthough cattle movement and commingling play an important role in the inter-herd transmission of pathogens, little is known about the effect of commingling of heifers at raising operations. The objective of this study was to compare the resistance ofE. coliand prevalence ofSalmonellafrom pooled faecal pats of heifers raised off-farm at multi-source raisers (MULTI) that raised heifers from at least two farms compared with on-farm raisers (HOME), with heifers from only that farm. MULTI faecal pat samples were collected from pens with animals that had arrived at the farm within the previous 2 months (AP) and from animals that would be departing the heifer raiser in 2–3 months (DP). Corresponding age sampling was conducted at HOME raisers. Odds of ampicillin resistance were 3·0 times greater inE. colicollected from MULTI compared to HOME raisers.E. colifrom AP pens had significantly (P< 0·05) higher odds of resistance to ampicillin, neomycin, streptomycin, and tetracycline compared to DP pens.Salmonellarecovery was not significantly different between heifer-raising systems (P= 0·3). Heifer-raising system did not have a major overall impact on selection of resistantE. coli, which was strongly affected by the age of the animals sampled.

The release of Escherichia coli into the environment from untreated manure can pose a threat to human health. Environmental survival of E. coli has been linked to extracellular fibers called curli. We investigated the effect of manure management (surface application followed by incorporation versus immediate incorporation) on the relative abundance of curli-producing E. coli in subsurface drainage effluent. Samples were collected from three dairy farms. The proportion of curli-producing E. coli in the manure storage facilities was uniform across the farms. However, the abundance of curli-producing E. coli was much greater (P < 0.05) in the tile drains of farms performing surface application of manure than in the tile drain of the farm that incorporated manure. This field observation was tested with controlled soil column experiments; the abundance of curli-producing E. coli in soil column effluents was greater (P < 0.05) when manure was surface-applied than when it was incorporated. Our findings suggest selection pressures resulting from the different manure application methods affected curli production by E. coli isolates transported through soil. Given the importance of curli production in pathogenesis, this work highlights the effect that manure management strategies may have on pathogenesis-associated phenotypes of bacteria in agricultural subsurface runoff.

Respiratory disease and diarrhea are the 2 most common diseases that result in the use of antimicrobial drugs in preweaned calves. Because the use of drugs in food animals, including dairy calves, has the potential for generating cross-resistance to drugs used in human medicine, it is vital to propose farm practices that foster the judicious use of antimicrobials while assuring animal health and productivity. The objective of this study was to use dairy farm calf treatment records to identify antimicrobial drug treatments in calves and to evaluate their effects on the prevalence of antimicrobial-resistant Escherichia coli from rectal swabs of preweaned dairy calves. Eight farms from central New York participated in the study, 3 farms using individual pen housing management and 5 farms using group pen housing management. Eligible study farms could not add antimicrobial drugs to the milk fed to preweaned calves and were required to have farm records documenting antimicrobial drug treatment of calves from birth to weaning. Three fecal E. coli isolates per calf were tested for susceptibility to 12 antimicrobial drugs using a Kirby-Bauer disk diffusion assay. A total of 473 calves were sampled, from which 1,423 commensal E. coli isolates were tested. Of the 9 antimicrobial drugs used on study farms, only enrofloxacin was significantly associated with reduced antimicrobial susceptibility of E. coli isolates, although treatment with ceftiofur was associated with reduced susceptibility to ceftriaxone. The median numbers of days from treatment with ceftiofur and enrofloxacin to rectal swab sampling of calves were 16 d (range: 1–39) and 12 d (range: 6–44), respectively. At the isolate level, treatment with enrofloxacin resulted in odds ratios of 2 [95% confidence interval (CI): 1–4] and 3 (95% CI: 2–6), respectively, for isolation of nonsusceptible E. coli to nalidixic acid and ciprofloxacin compared with calves not treated with enrofloxacin. Treatment with ceftiofur resulted in an odds ratio of 3 (95% CI: 0.9–12) for isolation of nonsusceptible E. coli to ceftriaxone compared with calves not treated with ceftiofur. Treatment with enrofloxacin resulted in selection of isolates that presented phenotypic resistance to both ciprofloxacin and ceftriaxone. Treatment with ceftiofur resulted in a higher prevalence of isolates resistant to ≥3 antimicrobial drugs (97%) compared with no treatment with ceftiofur (73%). These findings reinforce the necessity for continued implementation of practices at the dairy farm that support the sustainable and judicious use of antimicrobial drugs in dairy calves.

Nosocomial salmonellosis continues to pose an important threat to veterinary medical teaching hospitals. The objectives of this study were to describe an outbreak of salmonellosis caused by Salmonella enterica serovar Oranienburg within our hospital and to highlight its unique features, which can be used to help mitigate or prevent nosocomial outbreaks in the future. We retrospectively analyzed data from patients that were fecal culture-positive for Salmonella Oranienburg between January 1, 2006, and June 1, 2011, including historical, clinical, and pulsed-field gel electrophoresis (PFGE) data. Salmonella Oranienburg was identified in 20 horses, five alpacas, and three cows during this time frame, with dates of admission spanning the period from August, 2006, through January, 2008. We consider most of these patients to have become infected through either nosocomial or on-farm transmission, as evidenced by molecular subtyping results and supportive epidemiologic data. Interpretation of PFGE results in this outbreak was challenging because of the identification of several closely related Salmonella Oranienburg subtypes. Furthermore, a high percentage of cases were fecal culture-positive for Salmonella Oranienburg within 24 h of admission. These patients initially appeared to represent new introductions of Salmonella into the hospital, but closer inspection of their medical records revealed epidemiologic links to the hospital following the index case. Cessation of this outbreak was observed following efforts to further heighten biosecurity efforts, with no known cases or positive environmental samples after January, 2008. This study demonstrates that a Salmonella-positive culture result within 24 h of admission does not exclude the hospital as the source of infection, and it underscores the important role played by veterinary medical teaching hospitals as nodes of Salmonella infection that can promote transmission outside of the hospital setting.

Many of the drugs commonly used in lactating dairy cows result in residues in the milk, prohibiting its sale for human consumption. Milk withheld for sale because of drug treatment or from cows with high somatic cell counts is commonly called “waste milk.” One-third of dairy farms in the United States use waste milk to feed preweaned dairy calves. Limited information is currently available on the effect of this practice on the selection and dissemination of antibiotic-resistant bacteria. Pooled waste milk samples were collected from 34 dairy farms in central New York State with the objective of detecting the presence and quantity of drug residues in these samples. Samples were collected and refrigerated using ice packs and then stored at 4°C upon arrival at the Cornell laboratory (Ithaca, NY). Screening for β-lactam, tetracycline, and sulfonamide residues in the milk was performed using commercial enzyme-linked receptor-binding assay (SNAP) tests (Idexx Laboratories Inc., Westbrook, ME). Samples with a positive SNAP test were selected for screening using a multiresidue liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. The SNAP tests revealed that 75, 14.3, and 7.1% of waste milk samples (n = 34) contained β-lactam, tetracycline, and sulfamethazine residues, respectively. Of the samples sent for LC-MS/MS (n = 28), half had detectable quantities of drug residues. The most prevalent drugs detected by LC-MS/MS were ceftiofur (39.2%; mean ± SE concentration = 0.151 ± 0.042 μg/mL), penicillin G (14.2%; mean ± SE concentration = 0.008 ± 0.001 μg/mL), and ampicillin (7.1%; mean ± SE concentration = 0.472 ± 0.43 μg/mL). In addition, one sample had detectable concentrations of oxytetracycline and one sample had detectable concentrations of sulfadimethoxine. These results provide insight on drug residues present in waste milk from select farm in upstate New York, and additionally indicate the need for additional studies targeting on-farm treatments that could degrade drug residues present in waste milk and reduce the potential effects on the biosphere from the disposal and use of waste milk as a feed source.

Monitoring antimicrobial resistance trends among bacteria isolated from food animals and people is necessary to inform public policy regarding appropriate antimicrobial use. Our objectives were to describe the antimicrobial resistance status of Salmonella isolates from dairy cattle in the northeastern United States and to identify trends in resistance to various antimicrobial agents over time. Data were collected retrospectively for all bovine Salmonella isolates that were obtained from samples submitted to Cornell University's Animal Health Diagnostic Center between January 1, 2004 and December 31, 2011. Temporal trends in the prevalence of resistant Salmonella were investigated for each antimicrobial agent using the Cochran-Armitage trend test. Antimicrobial susceptibility testing was performed on 2745 bovine Salmonella isolates from clinical samples submitted during the study period. Overall resistance to each antimicrobial agent ranged from 0% (amikacin, ciprofloxacin, and nalidixic acid) to 72.0% (sulfadimethoxine). There was evidence of a significantly decreasing trend in prevalence of resistance to most agents: amoxicillin/clavulanic acid (AUG), ampicillin (AMP), cefoxitin (FOX), ceftiofur (TIO), ceftriaxone (AXO), chloramphenicol (CHL), chlortetracycline (CTET), florfenicol (FFN), kanamycin (KAN), neomycin (NEO), oxytetracycline (OXY), spectinomycin (SPE), streptomycin (STR), sulfadimethoxine (SDM), sulfisoxazole (FIS), and tetracycline (TET). Among the 265 isolates that were tested using the National Antimicrobial Resistance Monitoring System (NARMS) panel, the most common resistance patterns were pansusceptible (54.0%), AUG-AMP-FOX-TIO-AXO-CHL-KAN-STR-FIS-TET (18.1%), and AUG-AMP-FOX-TIO-AXO-CHL-STR-FIS-TET (12.1%). Increasing prevalence of S. enterica serovar Cerro over the course of the study period presumably had an impact on the observed resistance trends. Nevertheless, these results do not support the notion that the current level of antimicrobial use in dairy cattle is driving an increase in the emergence and dissemination of drug-resistant Salmonella in the region served by the laboratory.

OBJECTIVE To describe the antimicrobial resistance patterns of Salmonella isolates obtained from horses in the northeastern United States and to identify trends in resistance to select antimicrobials over time. SAMPLE 462 Salmonella isolates from horses. PROCEDURES Retrospective data were collected for all Salmonella isolates obtained from equine specimens that were submitted to the Cornell University Animal Health Diagnostic Center between January 1, 2001, and December 31, 2013. Temporal trends in the prevalence of resistant Salmonella isolates were investigated for each of 13 antimicrobials by use of the Cochran-Armitage trend test. RESULTS The prevalence of resistant isolates varied among antimicrobials and ranged from 0% (imipenem) to 51.5% (chloramphenicol). During the observation period, the prevalence of resistant isolates decreased significantly for amoxicillin—clavulanic acid, ampicillin, cefazolin, cefoxitin, ceftiofur, chloramphenicol, and tetracycline and remained negligible for amikacin and enrofloxacin. Of the 337 isolates for which the susceptibility to all 13 antimicrobials was determined, 138 (40.9%) were pansusceptible and 192 (57.0%) were multidrug resistant (resistant to ≥ 3 antimicrobial classes). The most common serovar isolated was Salmonella Newport, and although the annual prevalence of that serovar decreased significantly over time, that decrease had only a minimal effect on the observed antimicrobial resistance trends. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that current antimicrobial use in horses is not promoting the emergence and dissemination of antimicrobial-resistant Salmonella strains in the region served by the laboratory.

Although antimicrobial drugs are central to combat disease in modern medicine, the use of these drugs can have undesired consequences for human and animal health. One consequence is the post-therapy excretion of pharmacological agents, such as the elimination of drug residues at very low concentrations in the milk of lactating mammals. Limited information is currently available on the impact from the exposure of the gut microbiota to drug residues using in vivo natural models. The objective of our study was to address this knowledge gap and evaluate the effect on the fecal microbiota composition from feeding preweaned dairy calves raw milk with residual concentrations of ampicillin, ceftiofur, penicillin, and oxytetracycline from birth to weaning. At birth, thirty calves were randomly assigned to a controlled feeding trial where: 15 calves were fed raw milk with no drug residues (NR), and 15 calves were fed raw milk with drug residues (DR) by adding ceftiofur, penicillin, ampicillin, and oxytetracycline at final concentrations in the milk of 0.1, 0.005, 0.01, and 0.3 μg/ml, respectively. Fecal samples were rectally collected from each calf once a week starting at birth, prior to the first feeding in the trial (pre-treatment), until 6 weeks of age. Sequencing of the microbial 16S rRNA genes was conducted using the Illumina MiSeq, which provides a high resolution of the microbiota down to the genus level. Discriminant analysis showed that, except for pre-treatment samples, calves fed milk with drug residues and calves fed milk without drug residues easily discriminated at the genus level on their weekly microbial profile. However, analysis comparing the abundance of taxon between NR and DR showed significant differences only at the genus levels, and not at the phylum, class, order or family levels. These results suggest that although drug residues can result in clear discriminate gut microbial communities, they do not result in disruption of taxonomic levels above the genus.

The increasing size and complexity of US dairy farms could make it more difficult for a veterinary practitioner to effectively communicate protocol recommendations for prevention or treatment on the farm. A continuing education workshop was set up based on the results of research on dairy organizational communication on dairy farms, which resulted in a tool to assess dairy communication structure and flow. The workshop specifically focused on communication structure and whom to talk to when implementing health care changes in calf rearing. In addition, modern methods of veterinary–client communication knowledge and skills were provided. Primary outcomes of the workshops were to obtain feedback from participants about research findings and the communication model, to improve awareness about the complexity of communication structures on dairy farms, and to change participants' knowledge and skills associated with on-farm communication by providing communication theory and skills and an approach to evaluate and improve dairy organizational communication. Of the 37 participants completing the pre-program assessment, most recognized a need for themselves or their practice to improve communication with clients and farm employees. After the program, most participants were confident in their new communication skills and would consider using them. They highlighted specific new ideas they could apply in practice, such as conducting a “communication audit.” The results from the assessment of this communication workshop, focused on dairy veterinarians, highlighted the need for communication training in this sector of the profession and practitioners' desire to engage in this type of training.

The objective of this study was to identify patient symptoms and case outcomes that were more likely to occur as a result of Salmonella infections caused by bovine-associated subtypes (isolates that matched contemporary bovine isolates from New York by serovar and pulsed-field gel electrophoresis pattern), as compared to salmonellosis caused by non-bovine-associated subtypes. Data were collected in 34 counties of New York that comprise the Foodborne Diseases Active Surveillance Network (FoodNet) catchment area of the Centers for Disease Control and Prevention Emerging Infections Program. Patients with specimen collection dates between March 1, 2008 and March 1, 2010 were included. Symptoms and outcomes of 40 cases infected with bovine-associated Salmonella subtypes were compared to those of 379 control-cases infected with Salmonella isolates that were not bovine-associated. Cases were significantly more likely to have invasive salmonellosis (odds ratio, 3.8; p-value=0.02), after adjusting for age group, gender, and race. In addition, there was a marginal association between case status and the presence of blood in the stool (p-value=0.1) while ill. These findings might have implications for patient management, as a history of consuming undercooked foods of bovine origin or having direct contact with cattle in the few days prior to illness could be useful for suggesting a more proactive diagnostic approach as well as close monitoring for the need to implement more aggressive therapy.

Salmonella Cerro prevalence in US dairy cattle has increased significantly during the past decade. Comparison of 237 Salmonella isolates collected from various human and animal sources between 1986 and 2009 using pulsed- field gel electrophoresis, antimicrobial resistance typing, and spvA screening, showed very limited genetic diversity, indicating clonality of this serotype. Improved subtyping methods are clearly needed to analyze the potential emergence of this serotype. Our results thus emphasize the critical importance of population-based pathogen surveillance for the detection and characterization of potentially emerging pathogens, and caution to critically evaluate the adequacy of diagnostic tests for a given study population and diagnostic application.

Transmission of antimicrobial drug resistance from resistant bacteria to non-resistant strains is an important public health issue. In this study, we have examined the possibility of multiple resistance gene transfer between Escherichia coli and Salmonella in the natural setting. Bacteria isolated from calves concurrently shedding E. coli and Salmonella showed similar antimicrobial drug resistance patterns as measured by a broth dilution method. However, microarray analysis of the antibiotic resistance at the gene level revealed several differences in resistance gene profile. Resistance profiles of E. coli isolated from different farms were closer than the profile of E. coli and Salmonella isolated from the same farm. This shows that the chance of multiple resistance gene transfers between these species is unlikely.

Salmonella represents an important zoonotic pathogen worldwide, but the transmission dynamics between humans and animals as well as within animal populations are incompletely understood. We characterized Salmonella isolates from cattle and humans in two geographic regions of the United States, the Pacific Northwest and the Northeast, using three common subtyping methods (pulsed-field gel electrophoresis [PFGE], multilocus variable number of tandem repeat analysis [MLVA], and multilocus sequence typing [MLST]). In addition, we analyzed the distribution of antimicrobial resistance among human and cattle Salmonella isolates from the two study areas and characterized Salmonella persistence on individual dairy farms. For both Salmonella enterica subsp. enterica serotypes Newport and Typhimurium, we found multidrug resistance to be significantly associated with bovine origin of isolates, with the odds of multidrug resistance for Newport isolates from cattle approximately 18 times higher than for Newport isolates from humans. Isolates from the Northwest were significantly more likely to be multidrug resistant than those from the Northeast, and susceptible and resistant isolates appeared to represent distinct Salmonella subtypes. We detected evidence for strain diversification during Salmonella persistence on farms, which included changes in antimicrobial resistance as well as genetic changes manifested in PFGE and MLVA pattern shifts. While discriminatory power was serotype dependent, the combination of PFGE data with either MLVA or resistance typing data consistently allowed for improved subtype discrimination. Our results are consistent with the idea that cattle are an important reservoir of multidrug-resistant Salmonella infections in humans. In addition, the study provides evidence for the value of including antimicrobial resistance data in epidemiological investigations and highlights the benefits and potential problems of combining subtyping methods.

The objective of this study was to determine if the within-herd prevalence of fecal Salmonella shedding is higher in dairy herds with clinical outbreaks of disease, as compared to herds with subclinical infections only. Data were collected prospectively from dairy herds throughout New York that had at least 150 lactating cows and that received clinical service from participating veterinarians. After enrollment, Salmonella surveillance consisted of both environmental screening and disease monitoring within the herd. Herds positive by either environmental or fecal culture were sampled during three visits to estimate the within-herd prevalence of Salmonella. We characterized isolates by serovar and antimicrobial resistance pattern. Among 57 enrolled herds, 44 (77%) yielded Salmonella-positive samples during the study period; 27 (61%) of the positive herds had Salmonella isolated from environmental samples only, and 17 (39%) had one or more laboratory-confirmed clinical cases. The within-herd prevalence of fecal Salmonella shedding ranged from 0 to 53%. Salmonella Cerro was the predominant serovar, accounting for 56% of all isolates. Antimicrobial resistance ranged from zero to nine drugs, and 14 (32%) of the positive farms generated multidrug-resistant isolates. Herds with laboratory-confirmed clinical cases had a higher prevalence of fecal Salmonella shedding than herds that only generated positive environmental samples, as estimated by a Poisson regression model (prevalence ratio, 2.7; p = 0.01). An association between dairy herd outbreaks of salmonellosis and a higher prevalence of asymptomatic shedding should help guide strategies for reducing the public health threat of Salmonella, as the ability to recognize high-risk herds by clinical laboratory submissions presents an obvious opportunity to maximize food safety at the preharvest level. This is in contrast with other foodborne zoonotic pathogens, such as Campylobacter jejuni and Escherichia coli O157:H7, which occur widely in adult cattle without accompanying clinical disease.

The focus of this study was Salmonella enterica serotype Cerro, a potentially emerging pathogen of cattle. Our objectives were to document the within-herd prevalence of Salmonella Cerro among a sample of New York dairy herds, to describe the antimicrobial resistance patterns and pulsed-field gel electrophoresis types of the isolates, and to elucidate the status of this serotype as a bovine pathogen. Data were collected prospectively from dairy herds throughout New York that had at least 150 lactating cows and that received clinical service from participating veterinarians. Following enrollment, Salmonella surveillance consisted of both environmental screening and disease monitoring within the herd. Herds positive by either environmental or fecal culture were sampled during three visits to estimate the within-herd prevalence of Salmonella. Among 57 enrolled herds, 44 (77%) yielded Salmonella-positive samples during the study period. Of these, 20 herds (46%) were positive for Salmonella Cerro. Upon follow-up sampling for estimation of prevalence, Cerro was identified in 10 of the 20 herds; the median within-herd Cerro prevalence was 17%, with a maximum of 53%. Antimicrobial resistance ranged from zero to nine drugs, and eight (40%) of the Cerro-positive farms generated drug-resistant isolates. Eight XbaI pulsed-field gel electrophoresis types were represented among 116 isolates tested, although 89% of these isolates shared the predominant type. Among herds with clinical cases, cattle that had signs consistent with salmonellosis were more likely to test positive for Cerro than apparently healthy cattle, as estimated by a logistic regression model that controlled for herd as a random effect (odds ratio: 3.9). There is little in the literature concerning Salmonella Cerro, and published reports suggest an absence of disease association in cattle. However, in our region there has been an apparent increase in the prevalence of this serotype among cattle with salmonellosis. Other Salmonella serotypes important to bovine health have emerged to become leading causes of human foodborne disease, and close monitoring of Cerro is warranted.

Pulsed-field gel electrophoresis (PFGE) characterization of 335 temporally and spatially matched clinical, bovine, and human Salmonella enterica subsp. enterica isolates revealed 167 XbaI PFGE patterns. These isolates were previously classified into 51 serotypes and 73 sequence types, as determined by multilocus sequence typing. Discriminatory power of PFGE (Simpson's index, D = 0.991) was considerably higher than that of multilocus sequence typing (D = 0.920) or serotyping (D = 0.913). Although 128 PFGE types each only represented a single isolate, 8 PFGE types represented >4 isolates, including (i) three serotype Enteritidis and Heidelberg patterns that were only identified among human isolates, (ii) two PFGE patterns (each representing serotypes Bardo and Newport) that were significantly more common among bovine isolates as compared with human isolates; (iii) two PFGE types that each includes two serotypes (4,5,12:i:- and Typhimurium; Thompson and 1,7:-:1,5); and (iv) one PFGE type that includes eight Typhimurium isolates from humans and cattle. Characterization of isolates collected over multiple farm visits indicated that given specific PFGE types persisted over time on 11 farms. On an additional seven farms, isolates with a given sequence type represented multiple PFGE type, which typically only differed by

Aims: To investigate the genetic diversity among S. Enteritidis isolates from different geographic regions to evaluate the relationship between phage types (PTs) and variable number tandem repeat analysis (VNTR) loci. Methods and Results: We performed multiple-locus variable number tandem repeat analysis (MLVA) and phage typing on 245 S. Enteritidis isolates collected from sporadic human clinical cases in Michigan, Minnesota, New York, and Washington states between 2000 and 2007. Ninety-four MLVA types and 22 different PTs were identified. Specific PTs were associated with a predominant allele for certain VNTR loci. Cluster analysis using a minimum-spanning tree demonstrated two major clusters (I, II) and one minor cluster of isolates. PTs 8, 13a, 13 and 34 were significantly associated with MLVA cluster I. Phage types 1, 4, 6a, and 18 were significantly associated with MLVA cluster II. Conclusions: We found significant association between MLVA-based clusters and PTs. Certain VNTR loci were associated with specific PTs and could serve as useful molecular markers for S. Enteritidis in epidemiological investigations. Significance and Impact of the Study: MLVA genotyping in combination with phage typing can be used for effective characterization of S. Enteritidis isolates. It can also be useful for tracing possible sources during investigations of sporadic and outbreak cases of S. Enteritidis.

An intervention study was conducted to determine whether discontinuing the feeding of milk replacer medicated with oxytetracycline and neomycin to preweaned calves reduced antimicrobial resistance in Salmonella , Campylobacter , and Escherichia coli bacteria. Results demonstrated that the intervention did reduce multidrug resistance in these bacteria but that other factors also influenced multidrug resistance.

The objectives of this study were to determine the duration of fecal Salmonella shedding among dairy cattle in the northeastern United States following laboratory-confirmed clinical disease and to evaluate whether age group or serotype was associated with either shedding period or mortality. Study farms included 22 dairy herds that had at least two previous salmonellosis cases confirmed by fecal culture. Veterinarians continued to submit culture samples from clinical suspects following herd enrollment, and fecal samples from positive cattle were collected monthly until three sequential negative results were obtained or until loss to follow-up. There were 357 culture-positive clinical cases that each involved a single serotype during the shedding period. The Kaplan-Meier median duration of fecal Salmonella shedding was 50 days, and the maximum was 391 days. S. Newport was the predominant serotype, accounting for 51% of the cases. Age group and serotype were not significant predictors of Salmonella shedding duration in a Cox proportional hazards model, when stratifying by herd. However, the proportion of adult cows shedding for at least two consecutive monthly samples was significantly greater than the proportion of female calves shedding for this duration (Fisher’s exact test p-value < 0.01). Age group was also associated with mortality in this study; calves with salmonellosis were more likely to die than cows as estimated by a logistic regression model which controlled for herd as a random effect (p-value = 0.04).

Salmonella spp. are one of the most common causes of bacterial food-borne diseases worldwide (34). In the United States nontyphoidal Salmonella serotypes cause an estimated 1.4 million human salmonellosis cases, including approximately 550 deaths annually (27). Serotyping with the Kaufmann-White scheme is used commonly as a first step to differentiate Salmonella isolates. Serotyping of Salmonella isolates is based on lipopolysaccharide moieties on the cell surface (O antigens) and the flagellar proteins (H antigens), as well as capsular protein antigens (Vi antigen), which are only found in a few Salmonella serotypes (e.g., Salmonella enterica serotype Typhi). According to the Kaufmann-White scheme, Salmonella includes over 2,500 recognized serotypes (20). Many Salmonella bacteria are motile due to peritrichous flagella (28), which include a basal body, a propeller, and a hook. The motility of Salmonella depends on the rotation of the flagellar propeller (i.e., the filament), which includes either FliC (phase 1 antigen) or FljB (phase 2 antigen) flagellin (11). Most Salmonella serotypes, including Salmonella enterica serotype Typhimurium, are biphasic, meaning that they can express two distinct flagellar antigens (i.e., phase 1 and phase 2 antigens). Regulation of phase 1 and 2 antigen expression is under the control of the recombinase Hin. This recombinase facilitates inversion of a promoter element so that it either (i) transcribes fljB (which encodes the phase 2 antigen FljB) and fljA (which encodes a repressor of fliC, the gene encoding the phase 1 antigen FliC) (4, 37) or (ii) does not transcribe either of these genes. If this promoter is located in an orientation that does not allow for transcription of fljB and fljA, the lack of a repression of fliC transcription leads to expression of phase 1 flagellar antigens. Salmonella enterica serotype 4,5,12:i:− is a serotype that appears to be antigenically similar and genetically closely related to Salmonella serotype Typhimurium (which has the antigenic formula 4,5,12:i:1,2) but lacks expression of the second-phase flagellar antigen, which is 1,2 in Salmonella serotype Typhimurium (28). Salmonella serotype 4,5,12:i:− was the sixth most common Salmonella serotype among cases of human disease in the United States in 2006 (10) and the fourth most common serotype among human isolates in Spain in 1998 (18). Overall, the prevalence of Salmonella serotype 4,5,12:i:− among human cases has increased considerably in many countries in the world over the last 10 years (9, 10, 18, 29, 36). This Salmonella serotype has also been responsible for a number of human salmonellosis outbreaks over the last decades, including in Spain (1998), the United States (2004 and 2007), and Luxemburg (2006). Salmonella serotype 4,5,12:i:− has been isolated, particularly over the last decade, from a number of different foods and animals (1, 6, 13, 29, 38). While a number of separate studies, using molecular subtyping and characterization tools (e.g., genomic microarrays and PCR assays to test for gene presence/absence), have shown that Salmonella serotype 4,5,12:i:− isolates from Spain (15, 18) and the United States (1, 2, 38) are genetically closely related to Salmonella serotype Typhimurium, we are not aware of any comparative studies of Salmonella serotype 4,5,12:i:− isolates from Europe and the United States that have been published to date. In order to provide a better understanding of the transmission, ecology, and evolution of Salmonella serotype 4,5,12:i:−, we have assembled a collection of 190 Salmonella serotype 4,5,12:i:− and Typhimurium isolates from various sources and from two countries, the United States and Spain. These isolates were characterized by different molecular subtyping methods (i.e., multilocus sequence typing [MLST] and pulsed-field gel electrophoresis [PFGE]), followed by characterization of selected isolates for genomic deletions that may be responsible for the lack of phase 2 flagellum expression.

Objective—To determine the duration of fecal shedding of and serologic response to Salmonella spp after natural infection in dairy calves and characterize Salmonella organisms recovered from these herds. Design—Longitudinal study. Animals—Calves from 2 dairy herds (A and B) in the northeast United States that were identified at the beginning of a Salmonella outbreak. Procedures—Fecal samples were collected twice per week (herd A) or once per week (herd B); blood samples were collected for serologic testing once per week in both herds. Bacteriologic culture of fecal samples was performed, and Salmonella isolates were characterized by serotype, pulsed-field gel electrophoresis (PFGE) pattern, and antimicrobial resistance profile. Results—All Salmonella isolates from herd A were serovar Typhimurium var Copenhagen, had the same PFGE pattern, and were resistant to at least 9 antimicrobials. All isolates from herd B were Salmonella Typhimurium, represented 2 PFGE patterns, and were susceptible to all antimicrobials evaluated. The estimated duration of fecal shedding was 14 days in herd A and 9 days in herd B. Few calves were seropositive for antibody against Salmonella lipopolysaccharide within the first week after birth (0 of 20 in herd A and 13 of 79 in herd B) or seroconverted (6 in herd A and 4 in herd B). Fecal shedding was more common in calves that seroconverted, but overall, there was not a strong association between seropositivity and fecal shedding of Salmonella organisms. Conclusions and Clinical Relevance—Although the herds differed in serologic response and Salmonella subtype, the duration of fecal shedding among calves was similar between herds.

The objectives of this study were to estimate the incidence of salmonellosis among a large sample of dairy herds in the northeastern United States (both at the animal level and the herd level), to describe the serotypes and antimicrobial resistance profiles of the positive samples, and to determine whether various herd-level factors were important predictors of incidence. Participating veterinarians enrolled 831 dairy herds and submitted fecal samples from 2,565 female dairy cattle for Salmonella culture because of suspicion of clinical disease. Estimates of animal-level incidence rates were calculated for each age group as the number of cases per animal time at risk, and an estimate of herd-level incidence rate was calculated as the number of positive herds per herd time at risk. Descriptive analysis of serotype data and level of antimicrobial resistance was performed, and Poisson regression analysis was used to study associations between the within-herd incidence of salmonellosis and certain predictor variables (herd size, housing type, vaccination status, and prior history of Salmonella infection). Salmonella was isolated from 576 (22.5%) samples representing 93 herds. The animal-level incidence rates for preweaned female calves, heifers, and adult cows were 8.1, 0.04, and 1.8 cases per 1,000 animal-years, respectively. The herd-level incidence rate was 8.6 positive herds per 100 herd-years. Salmonella Newport was the predominant serotype, accounting for 41% of the cases, followed by Salmonella Typhimurium. Over 68% of all isolates were resistant to 5 or more antimicrobial agents. Herd size was the only significant predictor of the incidence of salmonellosis in a multivariable model; herds with at least 400 female dairy cattle had a higher incidence rate than smaller herds. Our results shed light on the impact of salmonellosis on the dairy industry in the northeastern United States, and they help clarify the role of dairy cattle as a source of Salmonella serotypes that are also important human pathogens.

Objective—To compare agreement between 2 pregnancy tests in dairy cattle. Design—Evaluation study. Animals—976 and 507 cattle for phases 1 and 2, respectively. Procedures—Blood samples were collected, and palpation per rectum (PPR) was performed on cattle. Blood samples for the pregnancy-specific protein B (PSPB) ELISA were sent by courier to a commercial laboratory with results returned later. Results of PPR were extracted from herd records. Statistical comparison of results was performed by use of a mixed linear model and N analysis. Results—Of 571 cattle classified as pregnant by the PSPB ELISA in phase 1, 30 (5%) were nonpregnant by PPR. Mean ± SE adjusted optical density (OD) of cattle classified pregnant by both tests was significantly higher (0.31 ± 0.01), compared with the adjusted OD of cattle classified pregnant by the PSPB ELISA and nonpregnant by PPR (0.22 ± 0.02). Of 255 cows classified pregnant by the PSPB ELISA in phase 2, 31 (12%) were nonpregnant by PPR. Mean ± SE adjusted OD of cattle classified pregnant by both tests was significantly higher (0.26 ± 0.01), compared with the adjusted OD of cattle classified pregnant by the PSPB ELISA and nonpregnant by PPR (0.21 ± 0.01). The N value was 0.82 and 0.81 for phases 1 and 2, respectively. Conclusions and Clinical Relevance—Good agreement existed between the 2 tests, especially at longer intervals after insemination. Discrepant results appeared to be attributable to a nonviable fetus, embryonic loss, or fetal loss.

In recent years, the proportion of Salmonella enterica infections represented by S. enterica serovar Newport has increased markedly among humans and animals. Multilocus variable-number tandem-repeat analysis (MLVA) has proven to be useful in discriminating other highly clonal Salmonella serovars. Here, we report on the development of a highly discriminatory MLVA for Salmonella serovar Newport.

Salmonella spp. represent one of the most common causes of bacterial foodborne illnesses around the world. The species Salmonella enterica contains more than 2500 serotypes, and emergence of new human pathogenic Salmonella strains and serotypes represents a major public health issue. Salmonella enterica subsp. enterica serotype 4,5,12:i:– represents a monophasic variant of Salmonella Typhimurium, which has rarely been identified before the mid-1990s. The prevalence of this serotype among human salmonellosis cases has increased considerably since the mid-1990s and Salmonella 4,5,12:i:– currently (i.e., the first decade of the 2000s) represents one of the most common serotypes among human cases in many countries around the world. This paper discusses our current knowledge of the global ecology, epidemiology, transmission, and evolution of this emerging Salmonella serotype.

A randomized intervention study was conducted to determine if discontinuing use of calf milk replacer medicated with oxytetracycline results in increased tetracycline susceptibility in Salmonella and Campylobacter spp. and Escherichia coli in dairy calves over a 12-month period. Dairy herds with enteric bacteria with known low tetracycline susceptibility were enrolled for the study. Fecal samples from preweaned calves and environmental samples were collected from eight dairy herds in Michigan and New York State. Samples were collected monthly for 3 months prior to and 12 months after four of the eight herds discontinued medicated milk replacer feeding. Salmonella and Campylobacter spp. and E. coli were isolated, and antimicrobial susceptibility testing was conducted using automated broth microdilution. A total of 804 intervention and 1,026 control calf fecal samples and 122 intervention and 136 control environmental samples were collected for testing. No differences in owner-reported morbidity and mortality between treatment groups were seen. The intervention was significantly associated with increasing tetracycline susceptibility in E. coli and Salmonella . Tetracycline susceptibility increased in intervention herds for the first 3 months after switching to nonmedicated milk replacer but declined in subsequent months. Discontinuing the practice of feeding medicated milk replacers to calves increased tetracycline susceptibility in E. coli and Salmonella on dairy farms, without increasing cattle disease, but declines in effectiveness after 3 months suggest that other factors contribute to decreasing susceptibility on the farm.

Objective—To determine the phase and quantitate the electromyographic (EMG) activity of the genioglossus, geniohyoideus, hyoepiglotticus, omohyoideus, sternohyoideus, sternothyroideus, and thyrohyoideus muscles of clinically normal horses during strenuous exercise. Animals—7 clinically normal adult horses (2 Thoroughbreds and 5 Standardbreds). Procedures—Bipolar electrodes were surgically implanted in the aforementioned muscles, and horses were subjected to an incremental exercise test on a high-speed treadmill. The EMG, heart rate, respiratory rate, and static pharyngeal airway pressures were measured during exercise. The EMG was measured as mean electrical activity (MEA). The MEA values for maximal exercise intensity (13 or 14 m/s) were expressed as a percentage of the MEA measured at an exercise intensity of 6 m/s. Results—MEA was detected during expiration in the genioglossus, geniohyoideus, sternohyoideus, and thyrohyoideus muscles and during inspiration in the hyoepiglotticus and sternothyroideus muscles. Intensity of the MEA increased significantly with exercise intensity in the genioglossus, geniohyoideus, and hyoepiglotticus muscles. Intensity of the MEA increased significantly in relation to expiratory pharyngeal pressure in the geniohyoideus and hyoepiglotticus muscles. Conclusions and Clinical Relevance—Once exercise intensity reached 6 m/s, no quantifiable additional increase in muscular activity was detected in the omohyoideus, sternohyoideus, sternothyroideus, and thyrohyoideus muscles. However, muscles that may affect the diameter of the oropharynx (genioglossus and geniohyoideus muscles) or rima glottis (hyoepiglotticus muscle) had activity correlated with the intensity of exercise or expiratory pharyngeal pressures. Activity of the muscles affecting the geometry of the oropharynx may be important in the pathophysiologic processes associated with nasopharyngeal patency.

SUMMARYThe objective of this study was to address the impact of heterogeneity of infectious period and contagiousness onSalmonellatransmission dynamics in dairy cattle populations. We developed three deterministic SIR-type models with two basic infected stages (clinically and subclinically infected). In addition, model 2 included long-term shedders, which were defined as individuals with low contagiousness but long infectious period, and model 3 included super-shedders (individuals with high contagiousness and long infectious period). The simulated dynamics, basic reproduction number (R0) and critical vaccination threshold were studied. Clinically infected individuals were the main force of infection transmission for models 1 and 2. Long-term shedders had a small impact on the transmission of the infection and on the estimated vaccination thresholds. The presence of super-shedders increasesR0and decreases the effectiveness of population-wise strategies to reduce infection, making necessary the application of strategies that target this specific group.

Objective—To evaluate effect of twin birth calvings on milk production, reproductive performance, and survival of lactating cows. Design—Retrospective observational cohort study. Animals—33,868 cows from 20 farms. Procedures—Data on age at calving for primiparous cows and mature equivalent milk yield for multiparous cows, assistance at calving, stillbirths, twin births, gestation duration, pregnancy at the end of the data collection period, and culling-death for all cows were extracted from farm computer records and used for statistical analysis. Results—Prevalence of twin parturitions was 1.3% (159/12,050) and 6.5% (1,410/21,818) for primiparous and multiparous cows, respectively. Primiparous and multiparous cows with singletons produced more milk than cows with live twins or at least 1 dead twin (primiparous, 33.1 vs 31.9 vs 31.2; multiparous, 36.5 vs 35.7 vs 35.0). Multiparous cows with dead twins produced less milk than cows with live twins. Compared with dams with singleton birth, cows with twins were 0.78 times as likely to conceive and 1.42 times as likely to die or be culled. Cows with dead twins also had increased time to conception, compared with live twins. Conclusions and Clinical Relevance—Twin birth was associated with decreased survival, milk production, and reproductive performance. Having at least 1 dead twin was even more detrimental than having live twins and resulted in decreased milk production and reproductive performance of lactating cows.

Nontyphoidal salmonellosis has been characterized by a pattern of dissemination of clonal Salmonella enterica. The most well-known example of this was S. enterica serovar Typhimurium definitive phage type 104 (DT104), a clone that emerged and disseminated globally in the 1990s (1). Other examples of epidemic clones include Salmonella serovar Wien, which spread from North Africa through Europe in the 1970s; S. Typhimurium phage type 10 (DT10), which disseminated across Canada in the 1970s; and other phage types of S. Typhimurium associated with cattle that disseminated widely in Europe during the 1980s (2). More recently, multidrug-resistant (MDR) S. Agona was disseminated in poultry in Belgium; MDR S. Paratyphi B dT+ in Germany, Belgium, and the Netherlands; and MDR S. Newport that harbors the plasmid-mediated AmpC resistance in the United States (3). Epidemic clones may remain largely restricted to the animal reservoir for years before their incidence rises among human infections: S. Typhimurium phage type 204c disseminated throughout the United Kingdom in cattle from 1980 through 1984, but its proportion among human S. Typhimurium infections was not high during that period (4). Some epidemic clones, for example DT10 and DT104, expanded in the absence of a specific resistance advantage (1). The mechanisms underlying emergence, dissemination, and subsequent decline of novel Salmonella clones are unknown, although some researchers have hypothesized that fitness-associated genetic factors may allow more efficient dissemination in specific hosts and environments (5) and that acquisition of these fitness genes may be phage mediated (6). One approach to understanding clonal replacement events is to conduct prospective surveillance of Salmonella infections in animal and human hosts. We initiated such a study to compare human- and animal-origin Salmonella strains by serovar, antimicrobial drug resistance pattern, and pulsed-field gel electrophoresis (PFGE).

This study compared the antimicrobial susceptibility of Shiga toxin-producing Escherichia coli (STEC) isolates from organic dairy farms, conventional dairy farms, and Minnesota county fairs. A total of 83 STEC isolates (43 O157 and 40 non-O157 STEC) were tested for antimicrobial susceptibility as determined by the automated broth microdilution method. Resistance to tetracycline was identified in 19 (23%) isolates and to sulphadimethoxine in 40 (48%) isolates. Half of the STEC isolates were resistant to at least one antimicrobial agent. Resistance to at least one antimicrobial agent was observed in 18 (62%) isolates from conventional farms and in 11 (48%) isolates from organic farms. Resistance to at least one antimicrobial agent was more frequent in isolates from calves (77%) than from cows (39%). Multidrug resistant patterns were more common in non-O157 STEC than O157 STEC. This study provides data to document the degree of STEC antimicrobial resistance from dairy cattle sources in Minnesota. The use of antimicrobial agents on farms, and other environmental influences, may affect resistance patterns in isolates from cattle sources. Systematic surveillance of STEC from cattle could potentially detect emergence of antimicrobial resistance that may be spread to humans through the food chain.

The objective of the study was to determine the effects of treatment with a controlled internal drug-release (CIDR) insert containing progesterone in a PGF2alpha-based presynchronization protocol on pregnancy rates at first service in lactating Holstein cows. A total of 1,318 (656 treatment and 662 control) cows from 5 farms were used in the analysis. Cows received a CIDR insert as part of the presynchronization protocol of 2 PGF2alpha injections given 14 d apart. The CIDR insert was applied during 7 d before the second PGF2alpha injection, whereas control cows received no CIDR insert. Serum progesterone concentrations were measured in samples collected at 37 +/- 3 d in milk (DIM; 7 d after the first PGF2alpha injection) and at 58 +/- 3 DIM, just before initiation of the Ovsynch protocol. According to serum concentrations of progesterone, cows were classified as having either high (or = 1 ng/mL) or low (1 ng/mL) progesterone. The proportion of cows with low progesterone at 37 +/- 3 DIM was similar for cows treated later with the CIDR insert (60.7%; n = 654) and for control cows (59.2%; n = 657). In contrast, use of the CIDR insert resulted in fewer low-progesterone cows (17.4%; n = 402) compared with control cows (30.6%; n = 399) at 58 +/- 3 DIM. No significant effect of the CIDR insert was detected on overall pregnancy rates. Pregnancy rates, as measured by the percentage of cows pregnant at 37 +/- 3 d post timed artificial insemination, for control cows having high or low progesterone at 58 +/- 3 DIM were 46.6 and 22.1%, respectively. For the CIDR group, pregnancy rates were 40.4 and 11.4%, respectively, for high- and low-progesterone cows at 58 +/- 3 DIM. Overall pregnancy rates were 36.4 and 34.5% for control cows and cows receiving the CIDR insert, respectively. A significant decreasing trend was observed in the proportion of cows having low progesterone as the body condition score increased, at 37 +/- 3 and 58 +/- 3 DIM. A significant increasing trend in the pregnancy rate was observed as body condition score increased. In conclusion, incorporation of CIDR inserts into a presynchronization protocol reduced the proportion of cows having low progesterone; however, the pregnancy rate did not differ between control cows and those receiving the CIDR insert. Earlier expression of estrus after the second PGF2alpha injection, and consequently improper timing of initiation of the Ovsynch protocol, could have negatively affected fertility in the CIDR-treated cows.

The objective of this study was to compare antimicrobial susceptibility of Salmonella isolated from conventional and organic dairy farms in the Midwest and Northeast United States. Environmental and fecal samples were collected from organic (n = 26) and conventional (n = 69) farms in Michigan, Minnesota, New York, and Wisconsin every 2 mo from August 2000 to October 2001. Salmonella isolates (n = 1,243) were tested using a broth microdilution method for susceptibility to amoxicillin-clavulanic acid, ampicillin, ceftiofur, ceftriaxone, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline, and trimethoprim-sulfamethoxazole. Herd-level logistic regression and logistic proportional hazards multivariable models were used to examine the association between farm management type and susceptibility to antimicrobial agents. For most antimicrobial agents tested, susceptibility of Salmonella isolates was similar on organic and conventional herds when controlling for herd size and state. Conventional farms were more likely to have at least one Salmonella isolate resistant to streptomycin using logistic regression (odds ratio=7.5; 95% confidence interval=1.7–5.4). Conventional farms were more likely to have Salmonella isolates with greater resistance to streptomycin (odds ratio=5.4; 95% confidence interval=1.5–19.0) and sulfamethoxazole (odds ratio=4.2; 95% confidence interval=1.2–14.1) using logistic proportional hazards models. Although not statistically significant, conventional farms tended to be more likely to have at least one Salmonella isolate resistant to 5 or more antimicrobial agents when compared with organic farms.

Insulin-like growth factor-I (IGF-I) is one of the most influential growth factors in cartilage repair. Maintenance of adequate IGF-I levels after articular repair procedures is complicated by the short biological half-life of IGF-I in vivo. This study investigated the potential for more prolonged IGF-I delivery through direct adenoviral mediated transduction of synovial tissues in the metacarpophalangeal (MCP) joints of horses. The use of a large animal model provided a structurally similar and metabolically relevant corollary to the human knee. The complete IGF-I coding sequence was packaged into an E1-E3 deleted adenovirus-5 vector under cytomegalovirus promoter control (AdIGF-I), and injected at varying total joint doses to the MCP joints of 14 horses. Direct injection of 20 and 50 x 10(10) AdIGF-I resulted in significant elevations of IGF-I in synovial fluid for approximately 21 days. Synovial tissue taken from injected joints at day 35 following injection and compared to tissue taken preinjection from the same joints revealed elevated synoviocyte IGF-I mRNA levels for the highest viral dose by in situ hybridization and real-time PCR techniques. AdIGF-I injections did not result in significant lameness, joint effusion or elevated total protein concentrations in the synovial fluid. Mild mononuclear infiltration of white blood cells was evident in histologic sections of the synovium in the second highest adenoviral IGF-I dose of 20 x 10(10) particles. Cartilage biopsies taken from all injected joints did not reveal any significant changes in proteoglycan levels nor in histological morphology, which included chondrocyte cloning, architecture, cell type or toluidine blue staining, when compared to control joints. Based on these findings, gene transfer of IGF-I to the synovium of joints can result in significant and persistent elevations of IGF-I ligand in synovial fluid with minimal detrimental effects. Direct IGF-I gene therapy may offer a simple approach in treating patients with acute cartilage injury.

Samples were collected from 26 organic and conventional farms and 12 county fairs in Minnesota during 2001 and 2002 to identify the presence of Escherichia coli O157. Immunomagnetic separation was used for isolation of E. coli O157. Isolates were further characterized by the presence of virulence marker genes (stx1, stx2, eaeA, E-hly, katP, etpD, and espP), antimicrobial susceptibility profiles, and genotypes. During 2001, E. coli O157 was isolated from 16 (5.2%) of 305 fecal samples and from 7 (36.8%) of 19 farms. During 2002, E. coli O157 was isolated from 6 (4.5%) of 132 fecal samples from weaned calves at 4 (23.5%) of 17 farms. During 2001 and 2002, cattle manure samples were collected from 12 county fairs, and E. coli O157 was isolated from 19 (11%) of 178 samples and 9 (75%) of 12 county fairs. Among 40 E. coli O157 isolates, 17 isolates (43%) had both the stx1 and stx2 genes, and 21 strains (53%) had the stx2 gene only. Thirteen percent of O157 isolates were resistant to tetracycline, and 25% were resistant to sulfadimethoxine. Heterogeneity of E. coli O157 strains was demonstrated by the presence of 22 different pulsed-field gel electrophoresis (PFGE) patterns. Four PFGE patterns matched those of isolates previously found in humans. The presence of E. coli O157 at county fairs suggests the potential for transmission to the public, who may have contact with cattle or their environment.

The objective of this study was to evaluate associations between herd-level factors and the isolation of Salmonella in calves from dairy farms in Minnesota, Wisconsin, Michigan and New York. Study farms were 129 conventional and organic farms enrolled without regard to previous history of Salmonella infection. Herds were sampled at 2-month intervals over a 1-year period. Salmonella was isolated in fecal samples from 176 (3.8%) of 4673 preweaned calves with 40 (31.0%) of 129 farms having at least one positive calf sample over the course of the study. Multivariable logistic regression using the generalized estimating equations approach was used to evaluate risk factors for Salmonella shedding after adjustment for effects of herd size, season, state of origin and the multiple sampling occasions per herd. Factors retained in the final model that were associated with an increased odds for Salmonella shedding were lack of routine feeding of milk replacer containing antimicrobials to preweaned calves (OR=2.8, 95% CI: 1.4, 5.8), use of maternity housing as a hospital area for sick cows more than once a month (OR=2.1, 95% CI: 1.1, 4.0), and cow prevalence level by visit, categorized into the following four-levels: > or =20% (OR=11.6, 95% CI: 5.7, 23.7), 10-19.9% (OR=4.7, 95% CI: 2.0, 11.5), 0.1-9.9% (OR=3.6, 95% CI: 1.5, 8.7) and 0% (reference level). Herd size was not associated with Salmonella shedding in the final multivariable model.