Economic impact of treatment of subclinical mastitis in early lactation using intramammary nisin.

Treatment of subclinical mastitis (SCM) during lactation is rarely recommended due to concerns related to both antimicrobial usage and costs associated with milk discard. Nisin is a naturally produced antimicrobial peptide with a Gram-positive spectrum that, when given to dairy cows, does not require milk discard. We evaluated the economic impact of treatment of SCM during early lactation using a nisin-based intramammary treatment under different scenarios which included various treatment costs, milk prices, and cure rates. We stochastically simulated the dynamics of SCM detected during the first week of lactation. The net economic impact was expressed in US dollars per case. The probabilities of an event and their related costs were estimated using a model that was based on pathogen-specific assumptions selected from peer-reviewed articles. Nisin cure rates were based on results of pivotal studies included in the FDA approval submission. Based on our model, the average cost of a case of intramammary infection (i.e., only true positive cases) in early lactation was $170 (90th percentage range = $148 to $187), while the cost of a clinical mastitis case was $521 (90th percentage range = $435 to $581). Both estimates varied with etiology, parity, and stage of lactation. When comparing the net cost of SCM cases (i.e., CMT-positive tests) detected during the first week of lactation, nisin treatment generated an average positive economic impact of $19 per CMT-positive case. Use of nisin to treat SCM was beneficial 93% of the time. Based on the sensitivity analysis, treatment would result in an economically beneficial outcome for 95% and 73% of multiparous and primiparous cows, respectively. At the herd level, use of intramammary nisin to treat SCM in cows in early lactation was economically beneficial in most tested scenarios. However, economic impact was highly influenced by factors such as rate of bacteriological cure, cost of treatment, and parity of affected animal. These factors should be considered when deciding to use nisin as a treatment for SCM.

Comparison of Minimum Inhibitory Concentrations of Selected Antimicrobials for Non-Aureus Staphylococci, Enterococci, Lactococci, and Streptococci Isolated from Milk Samples of Cows with Clinical Mastitis.

The objective of this study was to compare the minimum inhibitory concentrations of antimicrobials included in a commercial broth microdilution panel among Gram-positive pathogens that caused non-severe clinical mastitis on three Michigan dairy farms. Duplicate quarter milk samples were collected from eligible quarters of cows enrolled in a randomized clinical trial, cultured in a university laboratory, and identified using MALDI-TOF. Etiologies were grouped by genus as Enterococcus species (n = 11), Lactococcus species (n = 44), non-aureus Staphylococcus species (n = 39), or Streptococcus species (n = 25). Minimum inhibitory concentrations (MICs) were determined using the mastitis panel of a commercially available broth microdilution test. In vitro susceptibility was determined using approved guidelines and included breakpoints for mastitis pathogens, or when not available, breakpoints from other species. Most isolates were inhibited at or below breakpoints that demonstrated in vitro susceptibility. The proportions of susceptible isolates varied among pathogens for pirlimycin, penicillin, and tetracycline. The greatest proportion of resistance was observed for pirlimycin, tetracycline, and sulfadimethoxine. Survival analysis was performed to evaluate differences in MICs among pathogen groups. MIC values varied among pathogens for ceftiofur, cephalothin, erythromycin, penicillin, pirlimycin, and tetracycline. However, nearly all isolates were susceptible to ceftiofur and cephalothin, indicating that pathogen differences in MIC are not likely clinically relevant, as these are the two most commonly administered mastitis treatments in the United States. While differences in vitro susceptibility were observed for some antimicrobials, susceptibility was high to cephalosporin-based IMM treatments that are most commonly used and did not vary among pathogens.

A randomized control trial to test the effect of pegbovigrastim treatment at dry off on plasma and milk oxylipid profiles during early mammary gland involution and post-parturient period.

The early period of mammary gland involution is a critical juncture in the lactation cycle that can have significant effects on milk production and mammary gland health. Pegbovigrastim (PEG) administered 1 wk prior and on the day of parturition can enhance immune function and reduce the incidence of mastitis in the early postpartum period. Oxylipids are potent metabolites of polyunsaturated fatty acids (PUFA) and are important mediators of inflammation. The objective of this study was to evaluate effects of PEG given 1 wk before and at the day of dry-off (D0) on concentrations of oxylipids in plasma and milk from 7 d before D0 to 14 after, as well as the effects during the first 14 d of the subsequent lactation. We hypothesized that both pro- and anti-inflammatory oxylipids would vary based on initiation of mammary gland involution and that pegbovigrastim would affect oxylipid concentration, particularly those related to leukocytes. A complete randomized blocked design was used to enroll cows into either a PEG treatment group (n = 10) or control group (n = 10; CON). Blood samples were collected -7, -2, -1, 0, 1, 2, 4, 7, 14 d relative to dry-off and 5, 10, and 14 d post-calving. Samples were analyzed for PUFA and oxylipids in milk and plasma by ultra-performance mass-spectrometry and liquid chromatography tandem quadrupole mass spectrometry, respectively. Overall, 30 lipid mediators were measured both in milk and plasma. Repeated measures analyses revealed a significant interaction of treatment by time for milk 8-iso-keto-15-prostaglandin E2, prostaglandin F2α, plasma 8,12-iso-prostaglandin FαVI, 11-hydroxyeicosatetraenoic acid, and 12-hydroxyheptadecatienoic acid. The majority of milk PUFA and oxylipids differed significantly during early mammary gland involution and into the early postpartum period. This study demonstrated changes in oxylipids in milk secretions and plasma during early involution and further investigation may illuminate multiple complex processes and reveal targets for optimization of mammary gland involution.

Apparent prevalence and selected risk factors of methicillin-resistant Staphylococcus aureus and non-aureus staphylococci and mammaliicocci in bulk tank milk of dairy herds in Indiana, Ohio, and Michigan.

The purpose of this study was to determine the apparent prevalence and risk factors of methicillin-resistant Staphylococcus aureus and non-aureus staphylococci and mammaliicocci (NASM) in bulk tank milk (BTM) obtained from 300 dairy farms that belong to a cooperative collecting milk from Indiana, Michigan, and Ohio. Dairy field personnel recorded information about selected farm level risk factors and collected and froze BTM samples (n = 300) that were sent to Michigan State University researchers. Milk samples were thawed at room temperature and pre-enriched by adding 1 to 4 mL of Mueller-Hinton broth supplemented with 6.5% NaCl and incubated at 37°C for 24 h. Subsequently, 10 µL was plated on mannitol salt agar and Mueller-Hinton agar supplemented with 2.5% NaCl containing 2 mg/L oxacillin and 20 mg/L aztreonam. Colonies that grew on the selective media were subcultured on blood agar and identified using MALDI-TOF mass spectrometry. Phenotypic methicillin resistance was tested using cefoxitin disk diffusion. Conventional PCR was used to detect mecA and mecC in phenotypically resistant isolates. Of 550 isolates that were obtained from mannitol salt agar plates and 10 isolates from Mueller-Hinton agar plates, 16 species of NASM accounted for 84% of staphylococci, while S. aureus accounted for the remaining 16%. Among S. aureus, 4 isolates from 4 farms (1.3%) demonstrated phenotypic resistance to methicillin resistance but none carried mecA or mecC genes. Among NASM, 45 isolates from 40 farms (13.3%) demonstrated phenotypic resistance to methicillin. However, only 13 NASM isolates (7 Mammaliicoccus sciuri, 2 Staphylococcus haemolyticus, 1 Mammaliicoccus fleuretti, 1 Staphylococcus epidermidis, 1 Staphylococcus saprophyticus, and 1 Staphylococcus hyicus) from 13 farms were positive for mecA, whereas all were negative for mecC. Thus, the prevalence of mecA-positive NASM in BTM was 4.3%. Based on molecular results, this study demonstrated a low prevalence of methicillin resistance NASM from BTM samples collected from farms in the Upper Midwest. Dairy farms that contained ≤200 lactating cows and had swine located on the farm had a higher prevalence of methicillin-resistant NASM than smaller farms that did not contain swine.

Persistent effects of intramammary ceftiofur treatment on the gut microbiome and antibiotic resistance in dairy cattle.

BACKGROUND: Intramammary (IMM) ceftiofur treatment is commonly used in dairy farms to prevent mastitis, though its impact on the cattle gut microbiome and selection of antibiotic-resistant bacteria has not been elucidated. Herein, we enrolled 40 dairy (Holstein) cows at the end of the lactation phase for dry-cow therapy: 20 were treated with IMM ceftiofur (Spectramast®DC) and a non-antibiotic internal teat sealant (bismuth subnitrate) and 20 (controls) received only bismuth subnitrate. Fecal grab samples were collected before and after treatment (weeks 1, 2, 3, 5, 7, and 9) for bacterial quantification and metagenomic next-generation sequencing. RESULTS: Overall, 90% and 24% of the 278 samples had Gram-negative bacteria with resistance to ampicillin and ceftiofur, respectively. Most of the cows treated with ceftiofur did not have an increase in the number of resistant bacteria; however, a subset (25%) shed higher levels of ceftiofur-resistant bacteria for up to 2 weeks post-treatment. At week 5, the antibiotic-treated cows had lower microbiota abundance and richness, whereas a greater abundance of genes encoding extended-spectrum ß-lactamases (ESBLs), CfxA, ACI-1, and CMY, was observed at weeks 1, 5 and 9. Moreover, the contig and network analyses detected associations between ß-lactam resistance genes and phages, mobile genetic elements, and specific genera. Commensal bacterial populations belonging to Bacteroidetes most commonly possessed ESBL genes followed by members of Enterobacteriaceae. CONCLUSION: This study highlights variable, persistent effects of IMM ceftiofur treatment on the gut microbiome and resistome in dairy cattle. Antibiotic-treated cattle had an increased abundance of specific taxa and genes encoding ESBL production that persisted for 9 weeks. Fecal shedding of ESBL-producing Enterobacteriaceae, which was classified as a serious public health threat, varied across animals. Together, these findings highlight the need for additional studies aimed at identifying factors associated with shedding levels and the dissemination and persistence of antibiotic resistance determinants on dairy farms across geographic locations.

Invited review: Selective treatment of clinical mastitis in dairy cattle.

Treatment of clinical mastitis (CM) and use of antimicrobials for dry cow therapy are responsible for the majority of animal-defined daily doses of antimicrobial use (AMU) on dairy farms. However, advancements made in the last decade have enabled excluding nonsevere CM cases from antimicrobial treatment that have a high probability of cure without antimicrobials (no bacterial causes or gram-negative, excluding Klebsiella spp.) and cases with a low bacteriological cure rate (chronic cases). These advancements include availability of rapid diagnostic tests and improved udder health management practices, which reduced the incidence and infection pressure of contagious CM pathogens. This review informed an evidence-based protocol for selective CM treatment decisions based on a combination of rapid diagnostic test results, review of somatic cell count and CM records, and elucidated consequences in terms of udder health, AMU, and farm economics. Relatively fast identification of the causative agent is the most important factor in selective CM treatment protocols. Many reported studies did not indicate detrimental udder health consequences (e.g., reduced clinical or bacteriological cures, increased somatic cell count, increased culling rate, or increased recurrence of CM later in lactation) after initiating selective CM treatment protocols using on-farm testing. The magnitude of AMU reduction following a selective CM treatment protocol implementation depended on the causal pathogen distribution and protocol characteristics. Uptake of selective treatment of nonsevere CM cases differs across regions and is dependent on management systems and adoption of udder health programs. No economic losses or animal welfare issues are expected when adopting a selective versus blanket CM treatment protocol. Therefore, selective CM treatment of nonsevere cases can be a practical tool to aid AMU reduction on dairy farms.

Selective treatment of nonsevere clinical mastitis does not adversely affect cure, somatic cell count, milk yield, recurrence, or culling: A systematic review and meta-analysis.

Treatment of clinical mastitis (CM) contributes to antimicrobial use on dairy farms. Selective treatment of CM based on bacterial diagnosis can reduce antimicrobial use, as not all cases of CM will benefit from antimicrobial treatment, e.g., mild and moderate gram-negative infections. However, impacts of selective CM treatment on udder health and culling are not fully understood. A systematic search identified 13 studies that compared selective versus blanket CM treatment protocols. Reported outcomes were synthesized with random-effects models and presented as risk ratios or mean differences. Selective CM treatment protocol was not inferior to blanket CM treatment protocol for the outcome bacteriological cure. Noninferiority margins could not be established for the outcomes clinical cure, new intramammary infection, somatic cell count, milk yield, recurrence, or culling. However, no differences were detected between selective and blanket CM treatment protocols using traditional analyses, apart from a not clinically relevant increase in interval from treatment to clinical cure (0.4 d) in the selective group and higher proportion of clinical cure at 14 d in the selective group. The latter occurred in studies co-administering nonsteroidal anti-inflammatories only in the selective group. Bias could not be ruled out in most studies due to suboptimal randomization, although this would likely only affect subjective outcomes such as clinical cure. Hence, findings were supported by a high or moderate certainty of evidence for all outcome measures except clinical cure. In conclusion, this review supported the assertion that a selective CM treatment protocol can be adopted without adversely influencing bacteriological and clinical cure, somatic cell count, milk yield, and incidence of recurrence or culling.

Evaluation of reticuloruminal temperature for the prediction of clinical mastitis in dairy cows challenged with Streptococcus uberis.

Automated monitoring devices have become increasingly utilized in the dairy industry, especially for monitoring or predicting disease status. While multiple automated monitoring devices have been developed for the prediction of clinical mastitis (CM), limitations in performance or applicability remain. The aims of this study were to (1) detect variations in reticuloruminal temperature (RRT) relative to an experimental intramammary challenge with Streptococcus uberis and (2) evaluate alerts generated automatically based on variation in RRT to predict initial signs of CM in the challenged cows based on severity of clinical signs and the concentration of bacteria (cfu/mL) in the infected quarter separately. Clinically healthy Holstein cows without a history of CM in the 60 d before the experiment (n = 37, parity 1 to 5, ≥120 d in milk) were included if they were microbiologically negative and had a somatic cell count under 200,000 cells/mL based on screening of quarter milk samples 1 wk before challenge. Each cow received an intra-reticuloruminal automated monitoring device before the trial and was challenged with 2,000 cfu of Strep. uberis 0140J in 1 rear quarter. Based on interrupted time series analysis, intramammary challenge with Strep. uberis increased RRT by 0.54°C [95% confidence interval (CI): 0.41, 0.66] at 24 h after the challenge, which remained elevated until the end of the study. Alerts based on RRT correctly classified 78.3% (95% CI: 65.8, 87.9) of first occurrences of CM at least 24 h in advance, with a sensitivity of 70.0% (95% CI: 50.6, 85.3) and a specificity of 86.7% (95% CI: 69.3, 96.2). The accuracy of CM for a given severity score was 90.9% (95% CI: 70.8, 98.9) for mild cases, 85.2% (95% CI: 72.9, 93.4) for moderate cases, and 92.9% (95% CI: 66.1, 99.8) for severe cases. Test characteristics of the RRT alerts to predict initial signs of CM improved substantially after bacterial count in the challenged quarter reached 5.0 log10 cfu/mL, reaching a sensitivity of 73.5% (95% CI: 55.6, 87.1) and a specificity of 87.5% (95% CI: 71.0, 96.5). Overall, the results of this study indicated that RRT was affected by the intramammary challenge with Strep. uberis and the RRT-generated alerts had similar accuracy as reported for other sensors and algorithms. Further research that includes natural infections with other pathogens as well as different variations in RRT to determine CM status is warranted.

Incidence and Treatments of Bovine Mastitis and Other Diseases on 37 Dairy Farms in Wisconsin.

The aim of this research was to describe the incidence and treatments of mastitis and other common bovine diseases using one year of retrospective observational data (n = 50,329 cow-lactations) obtained from herd management software of 37 large dairy farms in Wisconsin. Incidence rate (IR) was defined as the number of first cases of each disease divided by the number of lactations per farm. Clinical mastitis (CM) remains the most diagnosed disease of dairy cows. Across all herds, the mean IR (cases per 100 cow-lactations) was 24.4 for clinical mastitis, 14.5 for foot disorders (FD), 11.2 for metritis (ME), 8.6 for ketosis (KE), 7.4 for retained fetal membranes (RFM), 4.5 for diarrhea (DI), 3.1 for displaced abomasum (DA), 2.9 for pneumonia (PN) and 1.9 for milk fever (MF). More than 30% of cows that had first cases of CM, DA, RFM, DI, and FD did not receive antibiotics. Of those treated, more than 50% of cows diagnosed with PN, ME and CM received ceftiofur as a treatment. The IR of mastitis and most other diseases was greater in older cows (parity ≥ 3) during the first 100 days of lactation and these cows were more likely to receive antibiotic treatments (as compared to younger cows diagnosed in later lactation). Cows of first and second parities in early lactation were more likely to remain in the herd after diagnosis of disease, as compared to older cows and cows in later stages of lactation. Most older cows diagnosed with CM in later lactation were culled before completion of the lactation. These results provide baseline data for disease incidence in dairy cows on modern U.S. dairy farms and reinforce the role of mastitis as an important cause of dairy cow morbidity.

Cows enrolled in a pilot randomized clinical trial evaluating pegbovigrastim administered to dairy cows at dry-off had reduced incidence of intramammary infection.

OBJECTIVE: The objective of this pilot study was to determine if an alternative dosing schedule of pegbovigrastim (PEG; Imrestor; Elanco Animal Health) affects mammary gland health, rear udder width, or milk production of healthy dairy cows. ANIMALS: 20 pregnant late-lactation Holstein cows in November 2019 through April 2020. PROCEDURES: Cows were randomly assigned to receive subcutaneous injections with either 15 mg of PEG (PEG group; n = 10) or a sham injection with saline (0.9% NaCl) solution (control group; 10) administered 7 days before dry-off and at dry-off. Quarter milk samples were collected for bacterial culture and somatic cells before and after dry-off and after calving. Mammary gland width was assessed before and after dry-off. Daily milk yields were evaluated after calving. RESULTS: The incidence of intramammary infection was 5 times greater for quarters of cows in the control group than for quarters of cows that received PEG. The effect of treatment on somatic cell count was not significant, but the effects of period and a treatment-by-period interaction were identified. Treatment did not significantly affect milk production in the subsequent lactation, but the effects of period and an interaction of treatment by period were identified. Rear udder width after dry-off was not significantly affected by treatment, but an effect of period was identified. CLINICAL RELEVANCE: In this pilot study, cows treated with PEG using an alternative dosing schedule had reduced incidence of intramammary infection and an interaction of treatment by sampling period was observed for milk yield. These results suggest that further studies with larger numbers of cows are warranted.

Realities, Challenges and Benefits of Antimicrobial Stewardship in Dairy Practice in the United States.

The use of antimicrobials for the treatment of food-producing animals is increasingly scrutinized and regulated based on concerns about maintaining the efficacy of antimicrobials used to treat important human diseases. Consumers are skeptical about the use of antibiotics in dairy cows, while dairy producers and veterinarians demonstrate ambivalence about maintaining animal welfare with reduced antimicrobial usage. Antimicrobial stewardship refers to proactive actions taken to preserve the efficacy of antimicrobials and emphasizes the prevention of bacterial diseases and use of evidence-based treatment protocols. The ability to broadly implement antimicrobial stewardship in the dairy industry is based on the recognition of appropriate antimicrobial usage as well as an understanding of the benefits of participating in such programs. The most common reason for the use of antimicrobials on dairy farms is the intramammary treatment of cows affected with clinical mastitis or at dry off. Based on national sales data, intramammary treatments comprise < 1% of overall antimicrobial use for food-producing animals, but a large proportion of that usage is a third-generation cephalosporin, which is classified as a highest-priority, critically important antimicrobial. Opportunities exist to improve the use of antimicrobials in dairy practice. While there are barriers to the increased adoption of antimicrobial stewardship principles, the structured nature of dairy practice and existing emphasis on disease prevention provides an opportunity to easily integrate principles of antimicrobial stewardship into daily veterinary practice. The purpose of this paper is to define elements of antimicrobial stewardship in dairy practice and discuss the challenges and potential benefits associated with these concepts.

The bovine milk microbiome - an evolving science.

Improved access to genome based, culture independent methods has generated great interest in defining the bovine milk microbiome. Several comprehensive reviews of this subject have recently been published and the purpose of this short review is to consolidate current understanding of the relevance and biological significance of this emerging topic. In contrast to mucosal organs that contain rich and well-characterized culturable and nonculturable microbial communities, milk obtained from the healthy bovine mammary gland usually contains few or no viable bacteria. The low bacterial biomass of milk has created methodological challenges that have resulted in considerable variability in results of studies that have used genomic methods to define the microbiota of milk obtained from healthy or diseased mammary glands. While genomes from several bacterial genera are routinely identified from samples of milk, teat skin and the teat canal, the viability, origin, and function of these organisms is uncertain as environmental factors have been shown to strongly influence the composition of these bacterial populations. Possible sources of microbial DNA include bacteria introduced from skin or the environment, bacteria trapped in teat canal keratin or bacteria engulfed by phagocytes. Researchers have not achieved consensus about key concepts such as the presence of a core commensal milk microbiome or dysbiosis as part of a causal pathway disrupting udder health. Understanding of the bovine milk microbiome has been greatly impeded by a lack of standardized methods used to collect, process, and assess bovine milk samples. Sample collection is a critical first step that will determine the validity of results. To minimize contamination with external sources of bacterial DNA, teat sanitation methods used for collection of milk samples that will be subjected to extraction and amplification of bacteria DNA should far exceed aseptic techniques used for collection of milk samples that will be submitted for microbiological culture. A number of laboratory issues have yet to be resolved. Contamination of low biomass samples with bacterial DNA from laboratory reagents is a well-known issue that has affected results of studies using bovine milk samples and results of sequencing of negative controls should always be reported. Replication of experiments has rarely been performed and consistency in results are lacking. While progress has been made, standardization of methods and replication using samples originating from differing farm conditions are critically needed to solidify knowledge of this emerging topic.

Wisconsin dairy farm worker perceptions and practices related to antibiotic use, resistance, and infection prevention using a systems engineering framework.

We studied farmworker practices and beliefs potentially contributing to transmission of bacteria and their associated antibiotic resistance genes (ARGs) among animals and farm workers to identify potential behavioral interventions to reduce the risk of bacterial transmission. Ten focus groups were conducted on eight Wisconsin dairy farms to assess potentially high-risk practices and farmworker knowledge and experiences with antibiotic use and resistance using the Systems Engineering in Patient Safety (SEIPS) framework. Farmworkers were asked to describe common on-farm tasks and the policies guiding these practices. We found workers demonstrated knowledge of the role of antibiotic stewardship in preventing the spread of ARGs. Worker knowledge of various forms of personal protective equipment was higher for workers who commonly reported glove-use. Additionally, workers knowledge regarding the importance of reducing ARG transmission varied but was higher than we had hypothesized. Programs to reduce ARG spread on dairy farms should focus on proper hand hygiene and personal protective equipment use at the level of knowledge, beliefs, and practices.

What Is Success? A Narrative Review of Research Evaluating Outcomes of Antibiotics Used for Treatment of Clinical Mastitis.

Treatment of clinical mastitis is the most common reason that antimicrobials are given to adult dairy cows and careful consideration of treatment protocols is necessary to ensure responsible antimicrobial stewardship. Clinical mastitis is caused by a variety of bacteria which stimulate an immune response that often results in spontaneous bacteriological clearance but can develop into long-term subclinical infections. Use of antimicrobial therapy is most beneficial for cases that are caused by pathogens that have a low rate of spontaneous cure but high rate of therapeutic cure. The purpose of this paper is to review studies that evaluated outcomes of antimicrobial therapy of clinical mastitis. Few studies reported differences in bacteriological cure among treatments and this outcome was rarely associated with clinical outcomes. Return to normal milk appearance was evaluated in most studies but demonstrated little variation and is not a reliable indicator of therapeutic success. Somatic cell count should be measured at quarter-level and will decline gradually after bacteriological clearance. Few researchers have evaluated important clinical outcomes such as post-treatment milk yield or culling. Few differences among approved antimicrobial therapies have been demonstrated and selection of antimicrobial therapy should consider the spectrum of activity relative to etiology.

Use of a systems engineering framework to assess perceptions and practices about antimicrobial resistance of workers on large dairy farms in Wisconsin.

We studied farmworker practices potentially contributing to transmission of bacteria and antimicrobial resistant genes (ARGs) among animals and farm workers to identify human behavioral interventions to reduce exposure risk. Ten focus groups were conducted on eight farms to explore potentially high-risk practices and farmworker knowledge and experiences with antimicrobial use and resistance using the Systems Engineering in Patient Safety (SEIPS) framework. Farmworkers were asked to describe common tasks and the policies guiding these practices. We found workers demonstrated knowledge of the role of antibiotic stewardship in preventing the spread of ARGs. Knowledge of various forms of personal protective equipment was higher for workers who commonly reported glove-use. Knowledge regarding the importance of reducing ARG transmission varied but was greater than previously reported. Programs to reduce ARG spread on dairy farms should focus on proper hand hygiene and personal protective equipment use but at the level of knowledge, beliefs, and practices.

A Cohort Study of the Milk Microbiota of Healthy and Inflamed Bovine Mammary Glands From Dryoff Through 150 Days in Milk.

The objective of this longitudinal cohort study was to describe the milk microbiota of dairy cow mammary glands based on inflammation status before and after the dry period. Individual mammary quarters were assigned to cohorts based on culture results and somatic cell count (SCC) at dryoff and twice in the first 2 weeks post-calving. Mammary glands that were microbiologically negative and had low SCC (< 100,000 cells/mL) at all 3 sampling periods were classified as Healthy (n = 80). Microbiologically negative mammary glands that had SCC ≥150,000 cells/mL at dryoff and the first post-calving sample were classified as Chronic Culture-Negative Inflammation (CHRON; n = 17). Quarters that did not have both culture-negative milk and SCC ≥ 150,000 cells/mL at dryoff but were culture-negative with SCC ≥ 150,000 at both post-calving sampling periods were classified as Culture-Negative New Inflammation (NEWINF; n = 6). Mammary glands with bacterial growth and SCC ≥ 150,000 cells/mL at all 3 periods were classified as Positive (POS; n = 3). Milk samples were collected from all enrolled quarters until 150 days in milk and subjected to microbiota analysis. Milk samples underwent total DNA extraction, a 40-cycle PCR to amplify the V4 region of the bacterial 16S rRNA gene, and next-generation sequencing. Healthy quarters had the lowest rate of PCR and sequencing success (53, 67, 83, and 67% for Healthy, CHRON, NEWINF, and POS, respectively). Chao richness was greatest in milk collected from Healthy quarters and Shannon diversity was greater in milk from Healthy and CHRON quarters than in milk collected from glands in the NEWINF or POS cohorts. Regardless of cohort, season was associated with both richness and diversity, but stage of lactation was not. The most prevalent OTUs included typical gut- and skin-associated bacteria such as those in the phylum Bacteroidetes and the genera Enhydrobacter and Corynebacterium. The increased sequencing success in quarters with worse health outcomes, combined with the lack of bacterial growth in most samples and the high PCR cycle number required for amplification of bacterial DNA, suggests that the milk microbiota of culture-negative, healthy mammary glands is less abundant than that of culture-negative glands with a history of inflammation.

Making Antibiotic Treatment Decisions for Clinical Mastitis.

Treatment of bovine mastitis is the most common reason that antibiotics are used for adult dairy cows. Although antibiotics are essential to treat severe cases of clinical mastitis, using antibiotics to treat many cases of nonsevere clinical mastitis does not result in improved bacteriologic or clinical outcomes. Mastitis treatment protocols should be pathogen specific; no antimicrobial therapy is recommended for many culture-negative or gram-negative cases when detected. Before withholding antibiotic therapy, it is important to assess the ability of affected cows to mount a successful immune response. When the immune system is compromised, antimicrobial therapy may be recommended.

Understanding the Milk Microbiota.

The milk microbiota is an intriguing area of research because milk with no bacterial growth in culture was long thought to be sterile. Recent DNA sequencing techniques have been developed that do not require bacteria to be culturable, and DNA from new bacteria have been reported in milk from dairy cow mammary glands with or without mastitis. Methodologies and results vary among research groups, and not enough is known about the milk microbiota for the results to be used for diagnosis or prognosis of mastitis.

Genetic variation in serum protein pattern and blood ß-hydroxybutyrate and their relationships with udder health traits, protein profile, and cheese-making properties in Holstein cows.

The aim of this study was to investigate in Holstein cows the genetic basis of blood serum metabolites [i.e., total protein, albumin, globulin, albumin:globulin ratio (A:G), and blood ß-hydroxybutyrate (BHB)], a set of milk phenotypes related to udder health, milk quality technological characteristics, and genetic relationships among them. Samples of milk were collected from 498 Holstein cows belonging to 28 herds. All animal welfare and milk phenotypes were assessed using standard analytical methodology. A set of Bayesian univariate and bivariate animal models was implemented via Gibbs sampling, and statistical inference was based on the marginal posterior distributions of parameters of concern. We observed a small additive genetic influence for serum albumin concentrations, moderate heritability (≥0.20) for total proteins, globulins, and A:G, and high heritability (0.37) for blood BHB. Udder health traits (somatic cell score, milk lactose, and milk pH) showed low or moderate heritabilities (0.15-0.20), whereas variations in milk protein fraction concentrations were confirmed as mostly under genetic control (heritability: 0.21-0.71). The moderate and high heritabilities observed for milk coagulation properties and curd firming modeling parameters provided confirmation that genetic background exerts a strong influence on the cheese-making ability of milk, largely due to genetic polymorphisms in the major milk protein genes. Blood BHB showed strong negative genetic correlations with globulins (-0.619) but positive correlations with serum albumin (0.629) and A:G (0.717), which suggests that alterations in the serum protein pattern and BHB blood levels are likely to be genetically related. Strong relationships were found between albumin and fat percentages (-0.894), between globulin and αS2-CN (-0.610), and, to a lesser extent, between serum protein pattern and milk technological characteristics. Genetic relationships between blood BHB and traits related to udder health and milk quality and technological characteristics were mostly weak. This study provides evidence that there is exploitable additive genetic variation for traits related to animal health and welfare and throws light on the shared genetic basis of these traits and the phenotypes related to the quality and cheese-making ability of milk.