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.

Effect of Holstein genotype on ex-vivo interleukin-1ß response to lipopolysaccharide (LPS), lipoteichoic acid (LTA) and heat-killed Gram-negative and Gram-positive bacteria.

Effects of Holstein genotype on interleukin-1ß response were assessed by ex-vivo stimulation of whole blood with lipopolysaccharide (LPS), lipoteichoic acid (LTA), or sonicated, heat-killed Gram-negative or Gram-positive bacteria. Holstein genotypes were unselected Holsteins (UH, n = 14) not subjected to selection pressures since the mid-1960s and contemporary Holsteins (CH, n = 13). Milk yield of UH and CH cows differ by more than 4500 kg/lactation. Whole blood was mixed with 0.01 µg LPS, 10 µg LTA or 2.5 × 106 CFU of sonicated, heat-killed E. coli, K. pneumoniae, S. marcescens, S. aureus, S. dysgalactiae, or S. uberis per mL of blood and incubated (4 h, 37 °C). Plasma IL-1ß was quantified by ELISA and log10-transformed concentrations analyzed with a multivariate linear mixed effects model. Responses to bacteria were greater than responses to LPS or LTA. Responses to LPS, LTA and the Gram-negative stimulants were greater in UH than in CH cows while responses to the Gram-positive bacteria did not differ between Holstein genotypes. In both genotypes, strong correlations were detected among IL-1ß responses to the Gram-negative stimulants and to LTA. There were strong correlations among IL-1ß responses to the Gram-positive bacteria in CH cows but only between S. aureus and S. dysgalactiae in UH cows. The IL-1ß response to S. uberis was highly correlated with responses to all of the Gram-negative stimulants in CH cows but only with E. coli in the UH cows. The reduced immune response could make contemporary cows more susceptible to infection by Gram-negative bacteria. Results confirm selection practices since the mid-1960s have altered immune response in the Holstein, at least to Gram-negative bacteria, and validate the need for additional studies to further evaluate the impacts of these selection practices on immune function in contemporary Holsteins.

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.

Improving farm-level antimicrobial stewardship benchmarks by reporting antimicrobial use within the context of both the magnitude of disease pressure and the outcome of therapy.

This manuscript explores a method of benchmarking antimicrobial use within the context of farm level therapeutic incidence (a proxy for disease incidence), and the outcome of that therapy. This is reported both within the same farm over time (2016-2019), as well as evaluated across participating farms. Reporting antimicrobial use in this format addresses multiple primary questions necessary for evaluating on farm antimicrobial stewardship: How much disease is recorded? How much antimicrobial use is recorded? How often are antimicrobials included in therapy for each disease? What is the outcome of therapy? The three primary metrics reported are: therapeutic events per 100 cow years (TE/100CY), antimicrobial regimens per 100 cow years (REG/100CY), and the percent therapeutic success (% Success). Success was defined as: the cow remained in the herd and had no further TE recorded within 30 days of the end of the TE being evaluated. These measures identify opportunities for change on an individual farm, such as improvement in disease prevention, or a change in choices about when to include an antimicrobial in the treatment protocol. Therapeutic outcomes provide additional context, in some instances demonstrating differences in recording practices and case definitions, while in other cases serving to safeguard animal welfare as efforts are made to decrease antimicrobial use in the future. Although developed for farm level reporting, the metrics may also be more broadly summarized to meet future reporting requirements for marketing chain or national level antimicrobial use reports. The process outlined here serves as a prototype to be considered when developing antimicrobial use reporting systems where farm level antimicrobial stewardship is the primary objective.

Effect of Holstein genotype on ex-vivo cytokine response to lipopolysaccharide (LPS) and lipoteichoic acid (LTA) during the periparturient period.

Effects of Holstein genotype on innate immune response were assessed with ex-vivo lipopolysaccharide (LPS) and lipoteichoic acid (LTA) stimulation of whole blood from unselected (UH, n = 10) and contemporary (CH, n = 11) Holsteins that differ in production by more than 4,500 kg/lactation. Blood was collected at -14, 7, 28, and 49 days in milk (DIM), mixed with a pathogen-associated molecular pattern (PAMP) molecule (0.01 or 1.0 µg LPS or 10 or 100 µg LTA per mL blood) and incubated (4 h, 37 °C). Plasma cytokines were quantified by ELISA, log10-transformed and analyzed by repeated measures with DIM as the repeated effect. Cytokine responses increased with PAMP dose and decreased as DIM increased. There was a genotype by LPS dose interaction for IL-1ß as response to the low dose was greater in UH but did not differ between genotypes for the high dose. The IL-1ß response was greater while the IL-6 response to LTA tended to be greater in UH than in CH cows. The more negative energy balance of CH cows did not impact genotype difference in cytokine responses. Results indicate selection since the mid-1960s has decreased ex-vivo, whole blood cytokine response of CH cows to LPS and to LTA.

The microbiome of common bedding materials before and after use on commercial dairy farms.

BACKGROUND: Bovine mastitis is one of the most economically important diseases affecting dairy cows. The choice of bedding material has been identified as an important risk factor contributing to the development of mastitis. However, few reports examine both the culturable and nonculturable microbial composition of commonly used bedding materials, i.e., the microbiome. Given the prevalence of nonculturable microbes in most environments, this information could be an important step to understanding whether and how the bedding microbiome acts as a risk factor for mastitis. Therefore, our objective was to characterize the microbiome composition and diversity of bedding material microbiomes, before and after use. METHODS: We collected 88 bedding samples from 44 dairy farms in the U.S. Unused (from storage pile) and used (out of stalls) bedding materials were collected from four bedding types: new sand (NSA), recycled manure solids (RMS), organic non-manure (ON) and recycled sand (RSA). Samples were analyzed using 16S rRNA sequencing of the V3-V4 region. RESULTS: The overall composition as well as the counts of several microbial taxa differed between bedding types, with Proteobacteria, Actinobacteria, Bacteroidetes and Firmicutes dominating across all types. Used bedding contained a significantly different microbial composition than unused bedding, but the magnitude of this difference varied by bedding type, with RMS bedding exhibiting the smallest difference. In addition, positive correlations were observed between 16S rRNA sequence counts of potential mastitis pathogens (bacterial genera) and corresponding bedding bacterial culture data. CONCLUSION: Our results strengthen the role of bedding as a potential source of mastitis pathogens. The consistent shift in the microbiome of all bedding types that occurred during use by dairy cows deserves further investigation to understand whether this shift promotes pathogen colonization and/or persistence, or whether it can differentially impact udder health outcomes. Future studies of bedding and udder health may be strengthened by including a microbiome component to the study design.

Prevalence, antibiotic resistance, virulence and genetic diversity of Staphylococcus aureus isolated from bulk tank milk samples of U.S. dairy herds.

BACKGROUND: Colonization of dairy cows by Staphylococcus aureus (S. aureus), especially those which are multi-drug resistant and toxin producing, is a concern for animal health and well-being as well as public health. The objective of this study was to investigate the prevalence, antibiotic resistance, gene content and virulence determinants of S. aureus in bulk tank milk samples (BTM) from U.S. dairy herds. RESULTS: BTM samples were collected, once in winter and once in summer, from 189 U.S. dairy herds. Of 365 BTM samples cultured, the sample and herd prevalence of S. aureus in BTM was 46.6% (170 of 365 samples) and 62.4% (118 of 189 herds), respectively. Among a subset of 138 S. aureus isolates that were stored for further analysis, 124 were genome sequenced after being confirmed as S. aureus using phenotypic tests. The most commonly identified antimicrobial resistance-associated gene was norA (99.2%) and mecA gene responsible for methicillin resistance (MRSA) was identified in one isolate (0.8%). The most frequently detected putative virulence genes were aur (100%), hlgB (100%), hlgA, hlgC, hlb (99.2%), lukE (95.9%) and lukD (94.3%). In the 53 staphylococcal enterotoxin positive isolates, sen (37.9%), sem (35.5%), sei (35.5%) and seg (33.1%) were the most frequently detected enterotoxin genes. Among the 14 sequence types (ST) and 18 spa types identified, the most common was ST2187 (20.9%) and t529 (28.2%), respectively. The most predominant clone was CC97 (47.6%) followed by CC unknown (36.3%). The single MRSA isolate belonged to ST72-CC8, spa type t126 and was negative for the tst gene but harbored all the other virulence genes investigated. CONCLUSION: Our findings indicated a high prevalence of S. aureus in BTM of U.S. dairy herds, with isolates showing little evidence of resistance to antibiotics commonly used to treat mastitis. However, isolates often carried genes for the various enterotoxins. This study identified predominant genetic clones. Despite lower prevalence, the presence of MRSA and multi-drug resistant strains in BTM poses a significant risk to animal and public health if their number were to increase in dairy environment. Therefore, it is necessary to continuously monitor the use of antibiotics in dairy cows.

Evaluation of the matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) system in the detection of mastitis pathogens from bovine milk samples.

MALDI-TOF is a chemistry analytical tool that has recently been deployed in the identification of microorganisms isolated from nosocomial environments. Its use in diagnostics has been extremely advantageous in terms of cost effectiveness, sample preparation easiness, turn-around time and result analysis accessibility. In the dairy industry, where mastitis causes great financial losses, a rapid diagnostic method such as MALDI-TOF could assist in the control and prevention program of mastitis, in addition to the sanitation and safety level of the dairy farms and processing facility. However, the diagnostic strengths and limitations of this test method require further understanding. In the present study, we prospectively compared MALDI-TOF MS to conventional 16S rDNA sequencing method for the identification of pathogens recovered from milk associated with clinical and subclinical bovine mastitis cases. Initially, 810 bacterial isolates were collected from raw milk samples over a period of three months. However, only the isolates (481) having both 16S rDNA sequencing and MALDI-TOF identification were included in the final phase of the study. Among the 481 milk isolates, a total of 26 genera (12 g-postive and 14 g-negative), including 71 different species, were taxonomically charecterized by 16S rDNA at the species level. Comparatively, MALDI-TOF identified 17 genera (9 g-positive and 8 g-negative) and 33 differernt species. Overall, 445 (93%) were putatively identified to the genus level by MALDI-TOF MS and 355 (74%) were identified to the species level, but no reliable identification was obtained for 16 (3.3%), and 20 (4.2%) discordant results were identified. Future studies may help to overcome the limitations of the MALDI database and additional sample preparation steps might help to reduce the number of discordances in identification. In conclusion, our results show that MALDI-TOF MS is a fast and reliable technique which has the potential to replace conventional identification methods for common mastitis pathogens, routinely isolated from raw milk. Thus it's adoption will strengthen the capacity, quality, and possibly the scope of diagnostic services to support the dairy industry.

Antimicrobial use quantification in adult dairy cows - Part 3 - Use measured by standardized regimens and grams on 29 dairies in the United States.

This study describes antimicrobial use in terms of standardized regimens per cow year (REG/CY) and grams per cow year (GMS/CY) for 29 dairies in the United States during the years 2016 and 2017. To explore potential priorities for antimicrobial stewardship programs, these measures were stratified by both disease syndrome and antimicrobial class. Potential confounders of use measurements are discussed and challenges for measure interpretation are identified. When measured as REG/CY, the results indicate that mastitis is the disease syndrome with the greatest contribution to overall antimicrobial use. However, when GMS/CY is measured, metritis, lameness and unknown disease syndromes are also significant contributors. When use is stratified by antimicrobial class, measures of REG/CY indicate the greatest magnitude of use is the cephalosporin class. However, when measures of GMS/CY are stratified by drug class, use within the penicillin class contributes more than any other single class. These differences highlight the need for a more complete understanding of the relationship between antimicrobial use measures and their relationship to antimicrobial resistance selection pressure.

Antimicrobial use quantification in adult dairy cows - Part 1 - Standardized regimens as a method for describing antimicrobial use.

This study describes a process to acquire and convert farm treatment records into a standardized regimen format. Multiple sources of on-farm data were utilized to convert the original treatment records to standardized regimens, enabling the generation of objective, granular descriptions of antimicrobial use. These standard regimen descriptions allow antimicrobial use data to be qualitatively stratified by active substance, disease syndrome treated, use category, and route of administration. Quantitative distributions are available for the grams of active substance per administration, the grams of active substance per regimen, the number of administrations, number of days of therapy and the time frame between the first and last administration. Granular quantification by this method informs future research, surveillance and policy.

Antimicrobial use quantification in adult dairy cows - Part 2 - Developing a foundation for pharmacoepidemiology by comparing measurement methods.

As stewardship programmes seek to influence therapeutic choices and reduce resistance selection pressure, it is essential to understand the multiple factors which may influence any given antimicrobial use measure. This article compares 9 measures of antimicrobial use pertaining to adult cows in US dairy systems based on treatment records standardized to a single standardized regimen (SReg) format. There are 3 fundamental parameters underlying the nine measures: amount of active substance (AMOUNT), number of SRegs (ANIMALS) and treatment time (TIME). Additionally, three use categories were considered in the analysis: control of mastitis (Dry Cow), treatment of detected mastitis (Clinical Mastitis) and treatment of other diseases (Other Treatment). The assumptions necessary for calculation of each of the nine measures are identified. The influence of measure choice on farm rank was explored at the farm level both within and across fundamental parameters. Across all use categories, when measures of TIME and ANIMALS were compared, correlations between measures were greater than 0.91. However, when measures of AMOUNT were compared to either measures of TIME or ANIMALS, the correlation between measures was more variable across use categories (R = 0.31-0.91). These comparisons demonstrate that the selection of antimicrobial use metric can affect which dairies are considered the highest users of antimicrobials. Measurement selection also influences which use category has the greatest potential for use reduction. This indicates that measure selection can influence which use category is prioritized as a target for reduction of antimicrobial use. Utilization of this information may allow those developing dairy antimicrobial stewardship programmes to better understand the potential effects of measurement selection on driving changes in antimicrobial use.

Colostrum Management for Dairy Calves.

Colostrum management is the single most important management factor in determining calf health and survival. Additional benefits of good colostrum management include improved rate of gain and future productivity. Successful colostrum management requires producers to provide calves with a sufficient volume of clean, high-quality colostrum within the first few hours of life. This article reviews the process of colostrogenesis and colostrum composition, and discusses key components in developing a successful colostrum management program. In addition, the article discusses approaches for monitoring and proposes new goals for passive immunity in dairy herds.

Use of Rapid Culture Systems to Guide Clinical Mastitis Treatment Decisions.

The selective treatment of clinical mastitis based on culture results can reduce antibiotic use by more than 50% without sacrificing treatment efficacy. Local laboratories reporting results in a 24-hour period, or adoption of on-farm milk culture systems, allow producers to make strategic treatment decisions. On-farm culture systems are most reliable when used to classify infections in broad diagnostic categories such as no growth, gram-positive, or gram-negative growth. Diagnostic accuracy is crucial for on-farm culture systems to be efficacious and economically advantageous. Quality assurance is necessary for the success of on-farm culture systems.

Association of bovine respiratory disease or vaccination with serologic response in dairy heifer calves up to three months of age.

OBJECTIVE: To investigate the association of bovine respiratory disease (BRD) or vaccination with serologic response in calves. ANIMALS: 94 Holstein calves. PROCEDURES: To assess the association between BRD and antibody titers, 38 calves < 3 months old that were treated for BRD were matched with 38 untreated calves. To investigate the effect of vaccination on antibody titers, 24 calves were randomly assigned to be vaccinated against bovine respiratory syncytial virus (BRSV), bovine viral diarrhea virus types 1 and 2, bovine herpesvirus type 1 (BHV1), and parainfluenza virus type 3 at 2 weeks of age (n = 6), 5 weeks of age (6), and both 2 and 5 weeks of age (6) or were assigned to be unvaccinated controls (6). Blood samples were obtained at I, 2, 5, and 12 weeks for determination of serum neutralization antibody titers against the vaccine viruses, bovine coronavirus, and Mannheimia haemolytica. Antibody rates of decay were calculated. RESULTS: Calves with initial antibody titers against BRSV < 1:64 that were treated for BRD had a slower rate of anti-BRSV antibody decay than did similar calves that were not treated for BRD. Calves with high initial antibody titers against BRSV and BHV1 had lower odds of BRD than did calves with low initial antibody titers against those 2 pathogens. Vaccination at 2 or 5 weeks of age had no effect on the rate of antibody decay. CONCLUSIONS AND CLINICAL RELEVANCE: Clinical BRD and the serologic response of dairy calves were associated with initial antibody titers against BRSV and BHV1. Serologic or clinical responses to viral exposure may differ in calves with low passive immunity.

Effect of delayed exposure of cattle to Mycobacterium avium subsp paratuberculosis on the development of subclinical and clinical Johne's disease.

OBJECTIVE: To evaluate the effect of delayed exposure of dairy cattle to Mycobacterium avium subsp paratuberculosis (MAP) on the incidence of those cows testing positive for MAP and developing clinical Johne's disease (CJD). ANIMALS: 79 cows not exposed to MAP as calves (unexposed cohort) and 260 cows exposed to MAP as calves (exposed cohort). PROCEDURES: Cows in the unexposed cohort were born into 5 MAP-uninfected herds and introduced at various ages into 5 MAP-infected herds where the exposed cohort cows were born and raised. Beginning when each cow was 24 months old, fecal and serum samples were collected annually from 2003 through 2006. Feces were cultured for MAP, and an ELISA was used to analyze serum samples for antibodies against MAP. Date and reason for culling were obtained from herd records. Incidence of positive culture and ELISA results and CJD was compared between unexposed and exposed cohort cows with Cox regression. RESULTS: Compared with exposed cohort cows, the hazard ratios for unexposed cohort cows having positive culture results, having positive ELISA results, and developing CJD were 0.12, 0.03, and 0.001, respectively, and those ratios increased by 2%, 6%, and 17%, respectively, for each month spent in an MAP-infected herd. CONCLUSIONS AND CLINICAL RELEVANCE: Delayed exposure of cows to MAP resulted in lower incidences of positive culture and ELISA results and CJD in those cows, compared with incidences of cows exposed to MAP since birth. The hazard of testing positive for MAP or developing CJD increased with time, regardless of cohort.

Is an individual calving pen better than a group calving pen for preventing transmission of Mycobacterium avium subsp paratuberculosis in calves? Results from a field trial.

The objective of this study was to quantify the efficacy of using individual calving pens (ICP) from which manure was removed between successive calving compared with group calving pens (GCP) for limiting transmission of Mycobacterium avium subsp paratuberculosis (MAP) in Holstein calves. Every other pregnant cow in three Minnesota MAP endemic herds was assigned to calve in either the ICP or the GCP within 48-72 h prior to expected calving. Heifer calves born in the ICP were assigned to the intervention group (n=238) while heifer calves born in the GCP were considered controls (n=211). Calves were separated from their dams as soon as was possible once the calf was found. The intervention within the ICP relative to the GCP was the removal of fecal material in the ICP immediately after each birth. Upon enrollment in 2005, calves were monitored into adulthood. Of the original animals enrolled, 318 were tested for MAP at least once in 2007, 2009, or 2010 using serum ELISA (ICP, n=165; GCP, n=141) and bacterial culture of feces (ICP, n=173; GCP, n=145) tests. Cox regression analysis was performed to evaluate the time until MAP test positivity. Cows born in the ICP had a hazard ratio of 0.37 (95% CI=0.34-0.4) for testing MAP serum ELISA positive, compared with cows born in GCP. Similarly, cows born in the ICP had a hazard ratio of 0.09 (95% CI=0.06-0.14) for testing MAP fecal culture positive, compared with cows born in GCP. The Cox proportional-hazard assumption was violated in both models such that differences observed in the instantaneous hazards of MAP positive outcomes between groups (ICP vs. GCP) subsequently diminished overtime. These findings indicate that using ICP for calving delays exposure to MAP in calves and provides an effective strategy for reducing peripartum MAP transmission risks in herds attempting to limit the impact of paratuberculosis.

Evaluation of the risk of paratuberculosis in adult cows fed Mycobacterium avium subsp paratuberculosis DNA-positive or -negative colostrum as calves.

OBJECTIVE: To estimate the risk of subclinical Mycobacterium avium subsp paratuberculosis (MAP) infection in cows that ingested MAP DNA-positive raw colostrum as calves, compared with risk in cows that ingested MAP DNA-negative raw colostrum as calves. ANIMALS: 205 calves born in 12 commercial dairy herds. PROCEDURES: Each calf was separated from its dam within 30 to 60 minutes after birth and fed raw colostrum. For each calf, samples of the colostrum fed were collected and tested for the presence of MAP DNA by use of a nested PCR assay for the target gene ISMAP02. Calves fed colostrum positive or negative for MAP DNA were classified into exposed (n = 69) and unexposed (136) groups, respectively. Each calf was tested for MAP infection at 30, 42, and 54 months of age by use of a serum ELISA and bacterial culture of feces. Weibull hazard regression models were used to evaluate the association between exposure to MAP DNA-positive colostrum and time to testing positive for MAP infection. RESULTS: Hazard of MAP infection was not different between groups (exposed vs unexposed) when serum ELISA, bacterial culture of feces, or both diagnostic tests (parallel interpretation) were positive. CONCLUSIONS AND CLINICAL RELEVANCE: Heifer calves fed MAP DNA-positive colostrum were at no greater risk of MAP infection, compared with heifer calves fed MAP DNA-negative colostrum. This result contradicts findings from other studies and should be interpreted with caution.

Evaluation of the association between fecal excretion of Mycobacterium avium subsp paratuberculosis and detection in colostrum and on teat skin surfaces of dairy cows.

OBJECTIVE: To evaluate the association between fecal excretion of Mycobacterium avium subsp paratuberculosis (MAP) by dairy cows in the periparturient period and detection of MAP DNA in colostrum specimens and on teat skin surfaces. DESIGN: Cross-sectional study. ANIMALS: 112 Holstein cows. PROCEDURES: Fecal specimens were collected within 48 to 72 hours prior to parturition, and colostrum and teat swab specimens were collected immediately after parturition. Detection of MAP in fecal specimens was performed via microbial culture, and detection of MAP DNA in colostrum and teat swab specimens was achieved via a PCR assay targeting the genetic element ISMAP02. Logistic regression was used to model the relationship between MAP fecal shedding status and detection of MAP DNA in colostrum or teat swab specimens. Population attributable fractions for the proportion of colostrum and teat swab specimens containing MAP DNA were also calculated. RESULTS: The odds of detecting MAP DNA in colostrum or teat swab specimens in cows with MAP-positive (vs negative) fecal specimens were 2.02 and 1.87 respectively. Population attributable fractions estimates suggested that withholding colostrum from MAP-positive cows could reduce the odds of exposing calves to MAP in colostrum by 18.2%. Not permitting natural suckling by calves could reduce the odds of exposing calves to MAP on the teat surfaces of MAP-positive cows by 19.5%. CONCLUSIONS AND CLINICAL RELEVANCE: Results underscored the need for strict adherence to practices that limit contact of calves with adult cows from the time of birth and promote hygienic colostrum handling to avoid possible contamination with MAP during colostrum harvest, storage, or feeding.

Effect of a plasma-derived colostrum replacement feeding program on adult performance and longevity in Holstein cows.

OBJECTIVE: To evaluate longevity, milk production, and breeding performance in adult Holstein cows fed either a plasma-derived commercial colostrum replacer (CR) or raw bovine maternal colostrum (MC) at birth. DESIGN: Randomized controlled clinical trial. ANIMALS: 497 heifer calves born in 12 commercial dairies located in Minnesota and Wisconsin. PROCEDURES: All calves were separated from their dams within 30 to 60 minutes after birth and systematically assigned to be fed either MC (control group [n = 261 calves]) or CR (treatment group [236]). Calves were observed from birth up to adulthood (approx 54 months old), during which time death and culling events plus milk yield and breeding performance data were collected. Time to death, time to culling, time to death or culling combined, time to first calving, and time to conception intervals were evaluated by use of proportional hazards survival analysis models. Number of times inseminated per conception and lifetime milk yield (up to 54 months old) were evaluated by use of general linear models. RESULTS: Cows fed CR as calves at the time of birth were no different than cows fed MC as calves with respect to overall risk of death, culling, or death or culling combined (from birth to 54 months of follow-up and from first calving to 54 months old); lifetime milk yield; and breeding performance. CONCLUSIONS AND CLINICAL RELEVANCE: No difference was detected in overall risk of death or culling, milk production, or reproductive performance between cows fed CR and those fed MC as calves at birth.

Experimental validation of a nested polymerase chain reaction targeting the genetic element ISMAP02 for detection of Mycobacterium avium subspecies paratuberculosis in bovine colostrum.

Colostrum samples experimentally inoculated with Mycobacterium avium subsp. paratuberculosis (MAP; strain K-10) at increasing concentrations between 1 x 10(1) and 1 x 10(9) cells/ml were tested for recovery of MAP DNA using a nested ISMAP02 target polymerase chain reaction initially developed for detecting MAP DNA in fecal samples. The following detection rates were achieved for sample replicates inoculated with unsonicated MAP pure stock: 100% between 1 x 10(7) and 1 x 10(9) cells/ml, 75% between 1 x 10(3) and 1 x 10(6) cells/ml, and 50% between 1 x 10(1)and 1 x 10(2) cells/ml replicates. Detection rates achieved for the colostrum sample replicates inoculated with sonicated MAP cell suspension were 75% for 1 x 10(9) cells/ml, 100% between 1 x 10(7) and 1 x 10(8) cells/ml, 75% for 1 x 10(6) cells/ml, 0 for 1 x 10(4) cells/ml, and 25% between 1 x 10(1) and 1 x 10(3) cells/ml. When negative control colostrum samples were tested, 16 of 18 (89%) samples were correctly detected as negative for MAP DNA using the current assay. In conclusion, the MAP DNA detection rates of the present assay improved with increasing concentrations of MAP in the colostrum sample replicates, although MAP DNA was also detected in 2 of 18 (11%) negative control samples, suggesting an undefined technical problem with the assay or, perhaps, sample contamination during preparation. Overall, the present findings suggest a potential role of the proposed polymerase chain reaction assay to detect MAP in colostrum. However, adoption of this test for use in routine screening of field colostrum for MAP awaits findings from an ongoing field validation study.