The history and future of the cornual nerve block for calf disbudding.

Disbudding damages the germinal horn bud cells and prevents subsequent horn growth in young calves. Hot-iron cautery or caustic paste are the most common disbudding techniques and are unequivocally painful procedures. An important technique in controlling the acute pain experienced during disbudding is the cornual nerve block (CNB) that uses a local anesthetic agent and targets a branch of the trigeminal cranial nerve, the zygomaticotemporal nerve, as it travels along the temporal groove of the skull. Though CNB have been used since 1932, practitioners have reported variability in achieving full desensitization of the horn bud region since its inception. This failure may have led to the establishment of variations in the CNB technique, without consensus on a reliable and repeatable approach. Reasons for CNB failures may include technical errors by the practitioner, such as an injection into subcutaneous or deep muscle bodies; biologically important path variations in the zygomaticotemporal nerve and its cornual branches; and secondary innervation of the horn bud region, particularly the cornual branches of the infratrochlear nerve. Further investigation into documenting and understanding CNB failure and alternatives, such as regional perfusion, is warranted.

Whole genome sequence analysis reveals high genomic diversity and potential host-driven adaptations among multidrug-resistant Escherichia coli from pre-weaned dairy calves.

Food-producing animals such as dairy cattle are potential reservoirs of antimicrobial resistance (AMR), with multidrug-resistant (MDR) organisms such as Escherichia coli observed in higher frequency in young calves compared to older cattle. In this study, we characterized the genomes of enteric MDR E. coli from pre-weaned dairy calves with and without diarrhea and evaluated the influence of host-level factors on genomic composition. Whole genome sequence comparative analysis of E. coli (n = 43) revealed substantial genomic diversity that primarily clustered by sequence type and was minimally driven by calf diarrheal disease status (healthy, diarrheic, or recovered), antimicrobial exposure, and dietary zinc supplementation. Diverse AMR genes (ARGs)-including extended-spectrum beta-lactamase genes and quinolone resistance determinants-were identified (n = 40), with unique sets of ARGs co-occurring in gene clusters with large AMR plasmids IncA/C2 and IncFIB(AP001918). Zinc supplementation was not significantly associated with the selection of individual ARGs in E. coli, however analysis of ARG and metal resistance gene pairs identified positive associations between certain aminoglycoside, beta-lactam, sulfonamide, and trimethoprim ARGs with acid, tellurium and mercury resistance genes. Although E. coli in this study lacked the typical virulence factors of diarrheagenic strains, virulence genes overlapping with those in major pathotypes were identified. Among the 103 virulence genes detected, the highest abundance and diversity of genes corresponded to iron acquisition (siderophores and heme uptake). Our findings indicate that the host-level factors evaluated in this study were not key drivers of genomic variability, but that certain accessory genes in enteric MDR E. coli may be enriched. Collectively, this work provides insight into the genomic diversity and host-microbe interface of MDR E. coli from pre-weaned dairy calves.

Impact of zinc supplementation on phenotypic antimicrobial resistance of fecal commensal bacteria from pre-weaned dairy calves.

The objective of this study was to evaluate the impact of dietary zinc supplementation in pre-weaned dairy calves on the phenotypic antimicrobial resistance (AMR) of fecal commensal bacteria. A repository of fecal specimens from a random sample of calves block-randomized into placebo (n = 39) and zinc sulfate (n = 28) groups collected over a zinc supplementation clinical trial at the onset of calf diarrhea, calf diarrheal cure, and the last day of 14 cumulative days of zinc or placebo treatment were analyzed. Antimicrobial susceptibility testing was conducted for Enterococcus spp. (n = 167) and E. coli (n = 44), with one representative isolate of each commensal bacteria tested per sample. Parametric survival interval regression models were constructed to evaluate the association between zinc treatment and phenotypic AMR, with exponentiated accelerated failure time (AFT) coefficients adapted for MIC instead of time representing the degree of change in AMR (MIC Ratio, MR). Findings from our study indicated that zinc supplementation did not significantly alter the MIC in Enterococcus spp. for 13 drugs: gentamicin, vancomycin, ciprofloxacin, erythromycin, penicillin, nitrofurantoin, linezolid, quinupristin/dalfopristin, tylosin tartrate, streptomycin, daptomycin, chloramphenicol, and tigecycline (MR = 0.96-2.94, p > 0.05). In E. coli, zinc supplementation was not associated with resistance to azithromycin (MR = 0.80, p > 0.05) and ceftriaxone (MR = 0.95, p > 0.05). However, a significant reduction in E. coli MIC values was observed for ciprofloxacin (MR = 0.17, 95% CI 0.03-0.97) and nalidixic acid (MR = 0.28, 95% CI 0.15-0.53) for zinc-treated compared to placebo-treated calves. Alongside predictions of MIC values generated from these 17 AFT models, findings from this study corroborate the influence of age and antimicrobial exposure on phenotypic AMR.

Effects of simplified group housing on behavior, welfare, growth performance, and health of preweaned dairy calves on a California dairy.

Most US dairy calves are raised in individual hutches for biocontainment purposes and to facilitate monitoring and handling of calves. However, individual hutches typically restrict calves' activity and social interactions. Previous studies showed that group housing (GH) is beneficial to calf welfare and is associated with social benefits. The adoption of GH on dairies is hindered by several concerns, with the primary concern being the potential for increased transmission of diseases due to heightened calf-to-calf contact. In light of this, our study aimed to compare the behavior, health, and growth outcomes of calves housed in groups of 3 to individually housed (IH) calves during the preweaning period. A total of 42 Holstein heifer calves on a commercial dairy in Northern California were enrolled in groups of 3 to different housing treatments; IH (n = 21) or GH (n = 21) between July and October 2020. Each treatment was composed of 7 groups of 3 calves each. Calves in the GH treatment were housed in groups of 3 from 6 to 10 d until 70 d of age. Individual pens consisted of one polyethylene hutch with a 1.5 m × 1.2 m outside exercise area. Group pens were constructed by assembling 3 polyethylene hutches with a 1.5 m × 3.6 m outside exercise area of wire panel fencing. Calves were weighed and measured for height at birth and weaning. Diarrhea and bovine respiratory disease (BRD) scores were recorded daily throughout the preweaning period. Cumulative incidence and hazard ratios were estimated for BRD and diarrhea for GH and IH. A mixed model with pen as a random effect was specified to evaluate the effect of treatment. Group-housed calves gained 0.64 ± 0.02 kg/d while IH calves gained 0.65 ± 0.02 kg/d. Similarly, there was no evidence for treatment differences in withers height gain in GH calves (0.22 ± 0.01 cm/d) compared with IH calves (0.21 ± 0.01 cm/d). The cumulative incidence of BRD based on the California scoring system in GH calves was 75 ± 9.68% compared with 66.66 ± 10.28% in IH calves. Group-housed calves had a BRD hazard of 1.14 times that of IH calves (95% CI: 1.21-2.40). The cumulative incidence of diarrhea (fecal score 3) in GH calves was 100% in comparison to 95.20% ± 4.66% in IH calves. The mean proportion of scan observations of calves feeding on concentrates was significantly higher in GH (0.145 ± 0.004/h) compared with IH calves (0.076 ± 0.003/h) during the preweaning period. The study results provide evidence that this simplified GH system provides benefits of GH without detrimental short-term effects on calf growth and health during the preweaning period.

Antimicrobial Susceptibility in Respiratory Pathogens and Farm and Animal Variables in Weaned California Dairy Heifers: Logistic Regression and Bayesian Network Analyses.

Weaned dairy heifers are a relatively understudied production group. Bovine respiratory disease (BRD) is the most common cause of antimicrobial drug (AMD) use, morbidity, and mortality in this production group. The study of antimicrobial resistance (AMR) is complicated because many variables that may affect AMR are related. This study generates hypotheses regarding the farm- and animal-level variables (e.g., vaccination, lane cleaning, and AMD use practices) that may be associated with AMR in respiratory isolates from weaned dairy heifers. A cross-sectional study was performed using survey data and respiratory isolates (Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni) collected from 341 weaned dairy heifers on six farms in California. Logistic regression and Bayesian network analyses were used to evaluate the associations between farm- and animal-level variables with minimum inhibitory concentration (MIC) classification of respiratory isolates against 11 AMDs. Farm-level variables associated with MIC classification of respiratory isolates included the number of source farms of a calf-rearing facility, whether the farm practiced onsite milking, the use of lagoon water for flush lane cleaning, and respiratory and pinkeye vaccination practices. Animal-level variables associated with a MIC classification included whether the calf was BRD-score-positive and time since the last phenicol treatment.

Unveiling the microbiome during post-partum uterine infection: a deep shotgun sequencing approach to characterize the dairy cow uterine microbiome.

BACKGROUND: The goal of this study was to assess the microbial ecology and diversity present in the uterus of post-partum dairy cows with and without metritis from 24 commercial California dairy farms using shotgun metagenomics. A set subset of 95 intrauterine swab samples, taken from a larger selection of 307 individual cow samples previously collected, were examined for α and ß diversity and differential abundance associated with metritis. Cows within 21 days post-partum were categorized into one of three clinical groups during sample collection: control (CT, n = 32), defined as cows with either no vaginal discharge or a clear, non-purulent mucus vaginal discharge; metritis (MET, n = 33), defined as a cow with watery, red or brown colored, and fetid vaginal discharge; and purulent discharge cows (PUS, n = 31), defined as a non-fetid purulent or mucopurulent vaginal discharge. RESULTS: All three clinical groups (CT, MET, and PUS) were highly diverse, with the top 12 most abundant genera accounting for 10.3%, 8.8%, and 10.1% of mean relative abundance, respectively. The α diversity indices revealed a lower diversity from samples collected from MET and PUS when compared to CT cows. PERMANOVA statistical testing revealed a significant difference (P adjusted < 0.01) in the diversity of genera between CT and MET samples (R2 = 0.112, P = 0.003) and a non-significant difference between MET and PUS samples (R2 = 0.036, P = 0.046). ANCOM-BC analysis revealed that from the top 12 most abundant genera, seven genera were increased in the natural log fold change (LFC) of abundance in MET when compared to CT samples: Bacteroides, Clostridium, Fusobacterium, Phocaeicola, Porphyromonas, Prevotella, and Streptococcus. Two genera, Dietzia and Microbacterium, were decreased in natural LFC of abundance when comparing MET (regardless of treatment) and CT, while no changes in natural LFC of abundance were observed for Escherichia, Histophilus, and Trueperella. CONCLUSIONS: The results presented here, are the current deepest shotgun metagenomic analyses conducted on the bovine uterine microbiome to date (mean of 256,425 genus-level reads per sample). Our findings support that uterine samples from cows without metritis (CT) had increased α-diversity but decreased ß-diversity when compared to metritis or PUS cows, characteristic of dysbiosis. In summary, our findings highlight that MET cows have an increased abundance of Bacteroides, Porphyromonas, and Fusobacterium when compared to CT and PUS, and support the need for further studies to better understand their potential causal role in metritis pathogenesis.

Management Practices of Bovine Mastitis and Milk Quality on Egyptian Dairies.

Milk production continues to be the main source of income for dairy producers, and mastitis continues to be the major health challenge for dairy cows worldwide. The objective of the current study was to describe the different management practices implemented in Egyptian dairies, which may influence mastitis and improve milk quality. An in-person survey was completed with herd managers and owners of 20 Egyptian dairies selected using a stratified random sample from four of Egypt's milk sheds. The questionnaire included 80 questions that inquired about herd demographics, mastitis status and control, milking practices, and management practices of the study dairies. Statistical analyses included descriptive statistics, multiple factor analysis (MFA), and hierarchical clustering to identify the important principal components and different dairy clusters. Of the surveyed herds, 69.50% had less than 500 lactating cows, with Holstein as the main cow breed. The reported average milk production on the study herds was 31.1 (SE ± 1.10) Kg/cow/day. Housing of lactating, dry, and close-up cows was in open lots. The majority of the dairies milked cows were three times per day (90.63%), the remaining milked cows four (5.00%) or a mix of four and three times per day. Furthermore, herringbone parlor design was the most common parlor design (66.79%) in the study dairies. The most common disinfectants used for both pre- and post-milking teat dip were iodine-based, 90.0% and 95.0%, respectively, while 52.16% of the dairies reported that their milkers wore gloves during milking. The reported mean annual percentage of mastitis was 52.3%, as a percent of the milking herd. The study dairies reported the inspection of udder health through either visual inspection (95.00%), udder hygiene score (86.88%), teat end score (71.88%), and/or using CMT (54.91%). Contagious mastitis pathogens were reported in 45.2% of the study herds. More than 50% of the study herds relied on importing pregnant cows or pregnant heifers as replacements. Multiple factor analysis identified 20 questions and represented 5 components of variability related to mastitis on dairies. The current survey of Egyptian dairies described the herd demographics and different management practices related to mastitis control and prevention. All the study dairies relied on the blanket intramammary antimicrobial drugs at dry off when current research elsewhere has identified selective dry cow therapy as an integral component of antimicrobial stewardship on dairies. Further research is required to identify the association between different management factors and the occurrence of mastitis.

Effect of dry cow therapy on antimicrobial resistance of mastitis pathogens post-calving.

The aim of this study was to evaluate the effect of dry cow therapy (DCT) on the antimicrobial resistance (AMR) profile of mastitis pathogens post-calving. A repository of isolates based on a DCT trial was utilized for the current study. A stratified random survey sample of cows from the trial were identified within the strata of season, herd, and trial treatment resulting in 382 cows. All isolates from the 382 cows were selected for the current study, which identified 566 isolates from milk samples collected at dry off (S1), post-calving (S2), and at the first clinical mastitis event up to 150 days in milk (S3). The AMR profiles were determined using broth microdilution method. Less than 10% of the coagulase-negative Staphylococcus species (CNS) isolates (n = 421) were resistant to tetracycline, ceftiofur, penicillin/novobiocin or erythromycin, while higher proportions of resistance to sulfadimethoxine (72%) and penicillin (28%) were observed. All Staphylococcus aureus (S. aureus) isolates (n = 4) were susceptible to all tested AMD except sulfadimethoxine, to which all isolates were resistant. Similarly, all Streptococcus spp. (n = 37) were susceptible to penicillin, penicillin/novobiocin, and ampicillin while resistant to tetracycline (17%). All coliforms (n = 21) were susceptible to ceftiofur, but resistance was recorded for sulfadimethoxine (70%), cephalothin (56%), and tetracycline (43%). The increased resistance percent from S1 to S2 was observed in CNS isolates from AMD-treated cows, with the highest increase recorded for penicillin (12.2%). Parametric survival interval regression models were used to explore the association between antimicrobial drug (AMD) therapy at dry off and the AMR phenotype post-calving. The accelerated failure-time metric was adopted to minimum inhibitory concentration measurements to permit interpretation of model exponentiated coefficients. Models for cows with CNS isolated at both S1 and S2 showed increased resistance against cephalothin, oxacillin, and ceftiofur in cows that received DCT from the same drug class, or a class with a shared resistance mechanism. In contrast, resistance of CNS isolates to tetracycline were associated with any AMD therapy at dry off. Resistance of CNS isolates to Penicillin decreased in CNS isolates in cows that received any AMD therapy at dry off compared to those that didn't. The study provided evidence that dry-cow IMM AMD was associated with AMR post-calving.

Effect of Group Housing of Preweaned Dairy Calves: Health and Fecal Commensal Antimicrobial Resistance Outcomes.

The objectives of this study were to investigate the effects of group housing (three calves per group) on bovine respiratory disease (BRD), diarrhea and antimicrobial resistance (AMR) to fecal commensal Escherichia coli (EC) and enterococci/streptococci (ES). Our study comprised two arms, one experimental and one observational. In the experimental arm, preweaned calves on a California dairy were randomized to either individual (IND; n = 21) or group (GRP; n = 21) housing, using a modified California-style wooden hutch. The study period lasted from birth to 56 days of age, during which calves were health scored daily. Cumulative incidence and hazard ratios were estimated for disease. Antimicrobial resistance outcomes were assessed using a prospective cohort design; feces were collected from each calf three times per week and EC and ES were evaluated for AMR using the broth microdilution method against a panel of 19 antimicrobial drugs (AMD). Analysis of treatment records was used to select calves that had been exposed (EXP) to an AMD-treated calf. In GRP, exposure occurred when a calf was a hutchmate with an AMD-treated calf. In IND, exposure occurred when a calf was a neighbor with an AMD-treated calf (TRT). Age-matched unexposed calves (UNEXP) were then selected for comparison. Proportions of AMR in fecal commensals among EXP, UNEXP, and TRT calves were compared between GRP and IND. Accelerated failure time survival regression models were specified to compare differences in minimum inhibitory concentration (MIC) of fecal commensals between EXP and UNEXP calves within each of GRP and IND calves separately. Group calves had a BRD hazard 1.94 times greater that of IND calves (p = 0.03), using BRD treatment records as the outcome. For AMR in EC isolates, higher resistance to enrofloxacin was detected in enrofloxacin-EXP GRP isolates compared with enrofloxacin-EXP IND isolates, and UNEXP GRP calves had lower resistance to ceftiofur compared with enrofloxacin-EXP and enrofloxacin-TRT calves. A significant housing-by-time interaction was detected for EC ceftiofur MIC in EXP GRP calves at 4-14 days post exposure to enrofloxacin (MIC EXP-UNEXP: µg/mL (95% CI): 10.62 (1.17, 20.07)), compared with UNEXP calves. The findings of this study show an increase in BRD hazard for group-housed calves and an increase in ceftiofur resistance in group-housed calves exposed to an enrofloxacin-treated calf.

Decision tree risk analysis for FMD outbreak prevention in Egyptian feedlots.

Foot and Mouth Disease (FMD) is one of the most contagious and economically influential viral diseases on animal health and productivity. Introduction of newly purchased animals to beef farms constitutes a greater risk for the FMD outbreaks in endemic countries. Physical examination of newly purchased animals in live animal markets and/or at the receiving farm, and the timing of preventative FMD vaccination may reduce the risk of FMD outbreaks under endemic conditions. Small (< 50 animals) and medium (50-500 animals) sized beef farms in Egypt constitute more than 60% of the country's beef industry where protocols for receiving newly purchased animals vary widely between farms. The current risk analysis aimed to build a decision tree model to reduce FMD outbreaks associated with introduction of replacement cattle on Egypt's medium sized beef farms. Management practices explored were the use of physical examinations and FMD vaccination and their timing for replacements with the goal of reducing losses due to FMD outbreaks. A producer survey revealed that more than 50% of the study herds relied on live animal markets as a source for replacements and reported more FMD outbreaks (P-value=0.09), FMD herd morbidity > 50% (p-value=0.05), and weight loss > 15 kg/animal in FMD clinical cases (P-value=0.01) in comparison to herds that received replacements from other farms, imported, or purchased from small stakeholders. More than 70% of the surveyed farms received replacements ≤ 1year old and reported significantly higher FMD outbreaks (P-value=0.02) in comparison to farms that received older animals. More than 80% of the surveyed farms performed physical examination of newly purchased animals before arrival at their premises. Of the surveyed farms, 73% reported FMD outbreaks with 67% of the outbreaks being reported during the Fall and Winter seasons. The decision tree identified physical examination of newly purchased animals prior to arrival and mixing with a premises beef herd followed by vaccination against FMD upon arrival as the intervention resulting in the lowest probability of FMD outbreak (8.9%). In contrast, herds that did not perform physical examination and delay the FMD vaccination for two or more weeks had the highest probability of FMD outbreaks (33.5%).

Effectiveness of Intramammary Antibiotics, Internal Teat Sealants, or Both at Dry-Off in Dairy Cows: Milk Production and Somatic Cell Count Outcomes.

Mastitis is the greatest disease challenge for dairy producers, with substantial economic impacts due to lost milk production. Amongst the approaches implemented to control and prevent mastitis on dairies are vaccination, pre- and post-milking teat dips, and treatments at dry-off including intramammary antibiotics and teat sealants. The objectives of our study were to evaluate the effect of different treatments at dry-off on the subsequent lactation's milk production and somatic cell count (SCC). A single-blinded controlled block randomized clinical trial was conducted between December 2016 and August 2018 on eight herds from four of the top ten milk-producing counties in California: Tulare, Kings, Stanislaus, and San Joaquin. The trial was repeated with cows enrolled during the winter and summer seasons to account for seasonal variability. Eligible cows were treated at dry-off with either intramammary antibiotics (AB), internal teat sealant (TS), AB + TS, or did not receive any treatment (control), and were followed through 150 days in milk (DIM) post-calving. The milk production and SCC data were extracted from monthly test day milk records (Dairy Comp 305, Valley Ag Software, Tulare, CA, USA). Two-piece spline linear mixed models were used to model the milk production (kg) and natural logarithm-transformed SCC. After accounting for parity, breed, season, and dry period duration, the milk model showed a significant increase in milk production (1.84 kg/day) in cows treated with AB + TS at dry-off in comparison to controls. There was no significant difference in the milk produced by cows that received either AB or TS (0.12 kg/day, and 0.67 kg/day, respectively) in comparison to the untreated cows. Different dry cow treatments were associated with a significant reduction in ln SCC during the first 150 DIM. The greatest reduction was associated with using AB + TS, followed by AB, and finally TS in comparison to controls. Dairies with high SCC may benefit from treating cows at dry-off with AB, TS, or both.

Evaluation of antimicrobial resistance and risk factors for recovery of intrauterine Escherichia coli from cows with metritis on California commercial dairy farms.

The goals of this study were to evaluate factors affecting recovery and antimicrobial resistance (AMR) in intrauterine E. coli in post-partum dairy cows with and without metritis from commercial California dairy farms. Using a cross-sectional study design, a total of 307 cows were sampled from 25 farms throughout California, from which a total of 162 intrauterine E. coli isolates were recovered. During farm visits, cows within 21 days post-partum were categorized in one of three clinical presentation groups before enrollment: metritis (MET, n = 86), defined as a cow with watery, red or brown colored, and fetid vaginal discharge; cows with purulent discharge (PUS, n = 106), defined as a non-fetid purulent or mucopurulent vaginal discharge; and control cows, (CTL, n = 115) defined as cows with either no vaginal discharge or a clear, non-purulent mucus vaginal discharge. Cows diagnosed as MET had significantly higher odds for recovery of E. coli compared to cows diagnosed as CTL (OR = 2.16, 95% CI: 1.17-3.96), with no significant difference observed between PUS and CTL, and PUS and MET. An increase in days in milk (DIM) at the time of sampling was significantly associated with a decrease in the odds ratio for E. coli recovery from intrauterine swabs (OR = 0.94, 95% CI: 0.89-0.98). All intrauterine E. coli were resistant to ampicillin (AMP), with an AMR prevalence of 30.2% and 33.9% observed for chlortetracycline and oxytetracycline, respectively. Only 8.6% of isolates were resistant to ceftiofur (CEFT), one of the most common drugs used to treat cows on farms sampled. No significant difference in the prevalence of AMR was observed among clinical groups at the individual cow level. At the farm level, a significantly higher odds for isolating intrauterine E. coli resistant to chlortetracycline (OR: 2.6; 95% CI: 3.7-58.0) or oxytetracycline (OR: 1.9; 95% CI: 1.4-33.8) was observed at farms that used an intrauterine infusion of oxytetracycline as a treatment for metritis when compared to those farms that did not use this practice. Findings from this study indicate the need for further research supporting a broader understanding of farm practices driving AMR in cows with metritis, as well as data to increase the accuracy of breakpoints for AMR classification of intrauterine E. coli from cattle.

Effectiveness of Intramammary Antibiotics, Internal Teat Sealants, or Both at Dry-Off in Dairy Cows: Clinical Mastitis and Culling Outcomes.

Intramammary antibiotic (AB) and internal teat sealants (TS) infusion at dry-off have been used to prevent intramammary infections (IMI) in dairy cows during the dry period and reduce the risk of mastitis during the dry period and subsequent lactation. A randomized clinal trial was completed on eight California dairy herds to estimate the effects of different dry cow therapies (AB, TS, AB + TS or None) on clinical mastitis and culling. A total of 1273 cows were randomized to one of the four treatment groups over summer and winter seasons. For each enrolled cow, microbiological testing was done on quarter milk samples collected from the first detection of clinical mastitis within the first 150 days in milk (DIM) in the subsequent lactation. Statistical analysis was done using generalized linear mixed models. There were no significant differences in the odds of clinical mastitis or culling between cows treated with AB, TS, or AB + TS compared to the controls. Dry cow therapy with AB and/or TS had no statistically significant effect on clinical mastitis and cow culling during the first 150 DIM.

Dynamic changes in fecal bacterial microbiota of dairy cattle across the production line.

BACKGROUND: Microbiota play important roles in the gastrointestinal tract (GIT) of dairy cattle as the communities are responsible for host health, growth, and production performance. However, a systematic characterization and comparison of microbial communities in the GIT of cattle housed in different management units on a modern dairy farm are still lacking. We used 16S rRNA gene sequencing to evaluate the fecal bacterial communities of 90 dairy cattle housed in 12 distinctly defined management units on a modern dairy farm. RESULTS: We found that cattle from management units 5, 6, 8, and 9 had similar bacterial communities while the other units showed varying levels of differences. Hutch calves had a dramatically different bacterial community than adult cattle, with at least 10 genera exclusively detected in their samples but not in non-neonatal cattle. Moreover, we compared fecal bacteria of cattle from every pair of the management units and detailed the number and relative abundance of the significantly differential genera. Lastly, we identified 181 pairs of strongly correlated taxa in the community, showing possible synergistic or antagonistic relationships. CONCLUSIONS: This study assesses the fecal microbiota of cattle from 12 distinctly defined management units along the production line on a California dairy farm. The results highlight the similarities and differences of fecal microbiota between cattle from each pair of the management units. Especially, the data indicate that the newborn calves host very different gut bacterial communities than non-neonatal cattle, while non-neonatal cattle adopt one of the two distinct types of gut bacterial communities with subtle differences among the management units. The gut microbial communities of dairy cattle change dramatically in bacterial abundances at different taxonomic levels along the production line. The findings provide a reference for research and practice in modern dairy farm management.

Bayesian estimation of sensitivity and specificity of a rapid mastitis test kit, bacterial culture, and PCR for detection of Staphylococcus aureus, Streptococcus species, and coliforms in bovine milk samples.

Our objectives were to evaluate the diagnostic accuracy of a rapid and novel immunochromatography-based mastitis kit that includes 3 independent tests to detect coliforms (Escherichia coli or Klebsiella pneumoniae), Streptococcus spp., and Staphylococcus aureus. The kit was developed to facilitate diagnostic-based mastitis treatment. Validation of the kit was based on 154 aseptically collected mastitis samples from 2 clinical herds (clinical population) and 120 milk samples from 3 nonclinical herds (nonclinical population) without clinical cases at the time of enrollment. One herd sampled at different times was common to both populations. A 3-test in 2-population Bayesian latent class model with uniform priors for all test parameters except specificity of culture, which was modeled informatively, was used to estimate sensitivity (Se) and specificity (Sp) of the test kit, culture, and PCR at the cow level. The mastitis test kit's 96.9% Sp for Streptococcus spp. had a low false positive percentage (3.1%), which, together with the kit's rapid turnaround time for results, makes it a suitable initial screening test that producers can use to identify clinical cows to treat based on Streptococcus spp. mastitis in kit-positive results. Due to the 60.4% kit Se, producers should follow up on Streptococcus spp. kit-negative cows using a confirmatory test such as PCR (Sp of 98.4%) or culture (Sp of 99.6%). In contrast, aerobic culture had Se of 76.5% and Sp of 99.6% for Streptococcus spp. Similarly, the Sp of the kit (98.2%) and culture (99.8%) for Staph. aureus were particularly high, and even though the kit's Se (61.0%) was lower than culture (88.4%; posterior probability of difference 98%), the kit could be beneficial before use of a confirmatory test for kit-negative samples due to its ease and rapid turnaround time. Mostly, quantitative real-time (q)PCR outperformed the kit's Se (37.7%) and Sp (92.9%) for coliforms, as well as the kit's Se (60.4%) for Streptococcus spp. However, qPCR may require more technical skills and turnaround time for final results. Use of the on-farm mastitis test kit evaluated in the present study could enhance sustainable antimicrobial drug use by rapidly identifying Streptococcus mastitis for targeted treatment. Furthermore, the kit may be used in a Staph. aureus outbreak where cows can be rapidly screened to identify cases for segregation or culling during an outbreak and kit-negative cows further confirmed by milk culture or qPCR. However, the cost-effectiveness of such an approach has not been investigated.

Factors Associated with Antimicrobial Stewardship Practices on California Dairies: One Year Post Senate Bill 27.

BACKGROUND: The current study is aimed at identifying the factors associated with antimicrobial drug (AMD) use and stewardship practices on conventional California (CA) dairies a year after CA Senate Bill 27. METHODS: Responses from 113 out of 1282 dairies mailed a questionnaire in 2019 were analyzed to estimate the associations between management practices and six outcomes including producer familiarity with medically important antimicrobial drugs (MIADs), restricted use of MIADs previously available over the counter (OTC), use of alternatives to AMD, changes in on-farm management practices, changes in AMD costs, and animal health status in dairies. RESULTS: Producers who reported having a veterinarian-client-patient relationship (VCPR) and tracking AMD withdrawal intervals had greater odds of being familiar with the MIADs. Producers who began or increased the use of preventive alternatives to AMD in 2019 had higher odds (OR = 3.23, p = 0.04) of decreased use of MIADs previously available OTC compared to those who did not. Changes in management practices to prevent disease outbreak and the use of diagnostics to guide treatment were associated with producer-reported improved animal health. In addition, our study identified record keeping (associated with familiarity with MIADs), use of alternatives to AMD (associated with management changes to prevent diseases and decreased AMD costs), and use of diagnostics in treatment decisions (associated with reported better animal health) as factors associated with AMD stewardship. CONCLUSIONS: Our survey findings can be incorporated in outreach education materials to promote antimicrobial stewardship practices in dairies.

Etiology and risk factors for bovine respiratory disease in pre-weaned calves on California dairies and calf ranches.

Our study objective was to estimate the magnitude of association of BRD risk factors including failure of passive immunity transfer, sex, age, and the detection of suspected BRD etiological pathogens in pre-weaned dairy calves in California. A conditional logistic regression model and a mixed-effects logistic regression model were used to estimate the association of these potential risk factors with BRD from a matched and nested case-control studies, respectively. For each exposure covariate, the odds ratio (OR) is the ratio of odds of an exposure in a BRD calf (case) to that in a non-BRD calf (control). In the matched case-control study, an interaction term between failure of transfer of passive immunity and sex of calf showed that female calves were more negatively impacted by failure of transfer of passive immunity compared to male calves. The odds ratios comparing failure of transfer of passive immunity in BRD score positive calves versus controls for male calves was 1.34 (95 % CI: 0.87, 2.06) and was 2.47 (95 % CI: 1.54, 3.96) for female calves. The model odds ratios varied from 1.74 (95 % CI: 1.26, 2.42) for Mycoplasma spp. to 9.18 (95 % CI: 2.60, 32.40) for Histophilus somni, with Mannheimia haemolytica and Pasteurella multocida having an OR of 6.64 (95 % CI: 4.39, 10.03) and 6.53 (95 % CI: 4.44, 9.59), respectively. For bovine respiratory syncytial virus positive calves, the OR was 4.60 (95 % CI: 3.04, 6.97). Findings from the nested case-control study showed that based on thoracic ultrasonography findings consistent with BRD, the odds of a calf being 1 day older compared to a day younger were 1.01 (95 % CI: 1.00, 1.02) among BRD cases. For the bacterial and viral pathogens, the OR for Mycoplasma spp. and Pasteurella multocida were 1.85 (95 % CI: 1.24, 2.75) and 1.86 (95 % CI: 1.28, 2.71), respectively. The OR values for these pathogens were similar when both thoracic auscultation and ultrasound findings were used to detect cases of BRD. Based on positive scores for BRD using the California BRD scoring system, the OR for facility type, calf ranch versus dairy farm, was 3.17 (95 % CI: 1.43, 7.01), Mannheimia haemolytica was 3.50 (95 % CI: 2.00, 6.11), Pasteurella multocida was 1.78 (95 % CI: 1.21, 2.60), and bovine coronavirus was 2.61 (95 % CI: 1.85, 3.70). Results from both study designs showed the difference in relative contributions of age, sex, immune status, and pathogens in BRD occurrence between cases and controls in pre-weaned dairy calves.

Estimating the Rates of Acquisition and loss of Resistance of Enterobacteriaceae to Antimicrobial Drugs in Pre-Weaned Dairy Calves.

The objective of this study was to investigate the effect of the antimicrobial drugs (AMD) on the shedding of resistant Enterobacteriaceae in feces of pre-weaned dairy calves. The AMD considered were ceftiofur, administered parenterally, and neomycin sulfate added in milk replacer and fed to calves during the first 20 days of life. Fifty-five calves, aged one to three days, were enrolled and followed to 64 days. Fecal samples were collected three times/week and treatments recorded daily. Enterobacteriaceae were quantified for a subset of 33 calves using spiral plating on plain, ceftiofur supplemented, and neomycin supplemented MacConkey agar. Negative binomial models were used to predict the association between treatment with AMD and the gain and loss of Enterobacteriaceae resistance over time. Acquisition of resistance by the Enterobacteriaceae occurred during treatment and peaked between days three to four post-treatment before decreasing to below treatment levels at days seven to eight post-treatment. Acquisition of neomycin resistance was observed on the first sampling day (day four from the start of feeding medicated milk replacer) to day eight, followed by cyclical peaks until day 29, when the Enterobacteriaceae counts decreased below pre-treatment. Enterobacteriaceae resistance against both AMD increased after AMD administration and didn't return to pre-therapeutic status until seven or more days after therapy had been discontinued. The study findings provide valuable insights into the dynamics of Enterobacteriaceae under routine AMD use in calves.

Antibiotic Resistance Genes and Associated Phenotypes in Escherichia coli and Enterococcus from Cattle at Different Production Stages on a Dairy Farm in Central California.

The objectives of this study were to characterize overall genomic antibiotic resistance profiles of fecal Escherichia coli and Enterococcus spp. from dairy cattle at different production stages using whole-genome sequencing and to determine the association between antimicrobial resistance (AMR) phenotypes and their corresponding genotypes. The Comprehensive Antibiotic Resistance Database (CARD) and ResFinder, two publicly available databases of antimicrobial resistance genes, were used to annotate isolates. Based on the ResFinder database, 27.5% and 20.0% of tested E. coli isolates (n = 40) harbored single and ≥3 antimicrobial resistance genes, respectively; for Enterococcus spp., we observed 87.8% and 8.2%, respectively. The highest prevalence of AMR genes in E. coli was for resistance to tetracycline (27.5%), followed by sulphonamide (22.5%) and aminoglycoside (20.0%); the predominant antimicrobial resistance genes in Enterococcus spp. targeted macrolide drugs (77.6%). Based on the CARD database, resistance to ≥3 antimicrobial classes was observed in all E. coli and 77.6% in Enterococcus spp. isolates. A high degree of agreement existed between the resistance phenotype and the presence of resistance genes for various antimicrobial classes for E. coli but much less so for isolates of Enterococcus. Consistent with prior work, fecal E. coli and Enterococcus spp. isolates from calves harbored a wide spectrum of resistance genes, compared to those from cattle at other production stages, based on the cross-sectional samples from the studied farm.

2019 Survey of Antimicrobial Drug Use and Stewardship Practices in Adult Cows on California Dairies: Post Senate Bill 27.

Antimicrobial resistance (AMR) is a global issue for both human and animal health. Antimicrobial drug (AMD) use in animals can contribute to the emergence of AMR. In January 2018, California (CA) implemented legislation (Senate Bill 27; SB 27) requiring veterinary prescriptions for medically important AMD use in food animals. The objective of our survey was to characterize AMD use, health management, and AMD stewardship practices of adult cows on CA dairies since the implementation of SB 27. In 2019, we mailed a questionnaire to 1282 California dairies. We received a total of 131 (10.2%) survey responses from 19 counties in CA. Our results showed that 45.6% of respondents included a veterinarian in their decision on which injectable AMD to purchase. Additionally, 48.8% of dairy producers included a veterinarian in their decision on which AMDs were used to treat sick cows. The majority (96.8%) of dairy producers were aware that all uses of medically important AMDs require a prescription. Approximately 49% of respondents agreed or strongly agreed that AMD use in livestock does not cause problems in humans. The survey documents antimicrobial use and stewardship practices in CA's dairy industry and focus areas for future research and education.