Relationships between milk culture results and milk yield in Norwegian dairy cattle

Associations between test-day milk yield and positive milk cultures for Staphylococcus aureus, Streptococcus spp., and other mastitis pathogens or a negative milk culture for mastitis pathogens were assessed in quarter milk samples from randomly sampled cows selected without regard to current or previous udder health status. Staphylococcus aureus was dichotomized according to sparse (≤1,500 cfu/mL of milk) or rich (>1,500 cfu/mL of milk) growth of the bacteria. Quarter milk samples were obtained on 1 to 4 occasions from 2,740 cows in 354 Norwegian dairy herds, resulting in a total of 3,430 samplings. Measures of test-day milk yield were obtained monthly and related to 3,547 microbiological diagnoses at the cow level. Mixed model linear regression models incorporating an autoregressive covariance structure accounting for repeated test-day milk yields within cow and random effects at the herd and sample level were used to quantify the effect of positive milk cultures on test-day milk yields. Identical models were run separately for first-parity, second-parity, and third-parity or older cows. Fixed effects were days in milk, the natural logarithm of days in milk, sparse and rich growth of Staph. aureus (1/0), Streptococcus spp. (1/0), other mastitis pathogens (1/0), calving season, time of test-day milk yields relative to time of microbiological diagnosis (test day relative to time of diagnosis), and the interaction terms between microbiological diagnosis and test day relative to time of diagnosis. The models were run with the logarithmically transformed composite milk somatic cell count excluded and included. Rich growth of Staph. aureus was associated with decreased production levels in first-parity cows. An interaction between rich growth of Staph. aureus and test day relative to time of diagnosis also predicted a decline in milk production in third-parity or older cows. Interaction between sparse growth of Staph. aureus and test day relative to time of diagnosis predicted declining test-day milk yields in first-parity cows. Sparse growth of Staph. aureus was associated with high milk yields in third-parity or older cows after including the logarithmically transformed composite milk somatic cell count in the model, which illustrates that lower production levels are related to elevated somatic cell counts in high-producing cows. The same association with test-day milk yield was found among Streptococcus spp.-positive pluriparous cows.     

Relationships between milk culture results and treatment for clinical mastitis or culling in Norwegian dairy cattle

In quarter milk samples from 2,492 randomly sampled cows that were selected without regard to their current or previous udder health status, the relationships between the following outcome variables were studied: treatment of clinical mastitis; the joint event of either treatment or culling for mastitis; culling for all reasons; culling specifically for mastitis; and the covariates of positive milk culture for Staphylococcus aureus, Streptococcus spp., and coagulase-negative Staphylococcus spp., or other pathogens, or of negative culture for mastitis pathogens. Microbiological diagnoses were assigned at the cow level, and altogether 3,075 diagnoses were related to the outcome variables. The relation between the absence of pathogens and rich (>1,500 cfu/mL of milk) or sparse (≤1,500 cfu/mL of milk) growth of Staph. aureus were also assessed separately for each outcome variable. The hazard of treatment of clinical mastitis was greater for cows diagnosed with Staph. aureus compared with cows with no pathogens in all analyses. Cows with sparse growth of Staph. aureus upon microbiological analysis were more likely to be treated for clinical mastitis, and cows with rich growth of the bacteria experienced a higher overall risk of culling when the models adjusted for cow composite milk somatic cell count. No difference between rich and sparse growth of Staph. aureus was found when mastitis was defined as the joint event of either culling for mastitis or treatment of clinical mastitis, and when the relationship with culling specifically for mastitis was assessed. The combined outcome of treatment and culling for mastitis was related to a positive diagnosis of Strep. spp. after cow composite milk somatic cell count was omitted from the model. Presence of Streptococcus spp. was also related to culling specifically for mastitis, whereas culling for all reasons and treatment of clinical mastitis was not related to a positive culture of Strep. spp. Presence of coagulase-negative Staph. spp. or other pathogens was not associated with either of the outcome variables.     

Postcalving udder health and productivity in cows approaching dry-off with intramammary infections caused by non-aureus Staphylococcus,Aerococcus, Enterococcus,Lactococcus, and Streptococcus species

The objective of this prospective cohort study was to explore associations between intramammary infection (IMI) in late-lactation cows and postcalving udder health and productivity. Cows (n = 2,763) from 74 US dairy herds were recruited as part of a previously published cross-sectional study of bedding management and IMI in late-lactation cows. Each herd was visited twice for sampling. At each visit, aseptic quarter milk samples were collected from 20 cows approaching dry-off (>180 d pregnant), which were cultured using standard bacteriological methods and MALDI-TOF for identification of isolates. Quarter-level culture results were used to establish cow-level IMI status at enrollment. Cows were followed from enrollment until 120 d in milk (DIM) in the subsequent lactation. Herd records were used to establish whether subjects experienced clinical mastitis or removal from the herd, and DHIA test-day data were used to record subclinical mastitis events (somatic cell count >200,000 cells/mL) and milk yield (kg/d) during the follow-up period. Cox regression and generalized estimating equations were used to evaluate the associations between IMI and the outcome of interest. The presence of late-lactation IMI caused by major pathogens was positively associated with postcalving clinical mastitis [hazard ratio = 1.5, 95% confidence interval (CI): 1.2, 2.0] and subclinical mastitis (risk ratio = 1.5, 95% CI: 1.3, 1.9). Species within the non-aureus Staphylococcus (NAS) group varied in their associations with postcalving udder health, with some species being associated with increases in clinical and subclinical mastitis in the subsequent lactation. Late-lactation IMI caused by Streptococcus and Streptococcus (Strep)-like organisms, other than Aerococcus spp. (i.e., Enterococcus, Lactococcus, and Streptococcus spp.) were associated with increases in postcalving clinical and subclinical mastitis. Test-day milk yield from 1 to 120 DIM was lower (?0.9 kg, 95% CI: ?1.6, ?0.3) in late-lactation cows with any IMI compared with cows without IMI. No associations were detected between IMI in late lactation and risk for postcalving removal from the herd within the first 120 DIM. Effect estimates reported in this study may be less than the underlying quarter-level effect size for IMI at dry-off and postcalving clinical and subclinical mastitis, because of the use of late-lactation IMI as a proxy for IMI at dry-off and the use of cow-level exposure and outcome measurements. Furthermore, the large number of models run in this study (n = 94) increases the chance of identifying chance associations. Therefore, confirmatory studies should be conducted. We conclude that IMI in late lactation may increase risk of clinical and subclinical mastitis in the subsequent lactation. The relationship between IMI and postcalving health and productivity is likely to vary among pathogens, with Staphylococcus aureus, Streptococcus spp., Enterococcus spp., and Lactococcus spp. being the most important pathogens identified in the current study.     

Association between Mycobacterium avium subspecies paratuberculosis infection and milk production in two California dairies

The association between Mycobacterium avium ssp. paratuberculosis (MAP) and milk production was estimated on 2 California dairies using longitudinal data from 5,926 cows. Both study herds had moderate MAP seroprevalence, housed cows in freestalls, and had Johne's disease control programs. Cow MAP status was determined using both serum ELISA and fecal culture results from cows tested at dry-off and from whole-herd tests. Potential confounders were evaluated based on a causal diagram. Mixed models with 2 functions (splines) for days in milk (DIM) representing milk production pre- and postpeak used in similar studies were further modified to use each cow's observed DIM at peak and lactation length. Cows that were seropositive produced 2.5 kg less 4% fat-corrected milk (FCM) per day than their seronegative herdmates. In addition, cows that were fecal-culture positive by liquid culture and confirmed by PCR produced 2.2 kg less 4% FCM per day than their fecal-culture negative herdmates. The decrease in milk production in MAP test-positive compared with test-negative cows started in the second lactation. A switch in MAP status in either ELISA or fecal culture results from positive to negative had no significant association with milk production. Modified DIM functions that used the observed DIM at peak had better model fit than another function that assumed a fixed peak at 60 DIM. Cows that tested positive for MAP on serum ELISA or fecal culture produced less milk than cows that tested negative, and the association between MAP and milk production was not confounded by mastitis, elevated somatic cell counts, or uterine or metabolic cow conditions.     

Milk production change following clinical mastitis and reproductive performance compared among J5 vaccinated and control dairy cattle

Naturally occurring cases of bovine clinical mastitis (CM) were studied among J5 vaccinates and controls on 3 commercial dairy farms. Milk production change and reproductive performance following CM were compared between the 2 groups. Among 306 controls and 251 vaccinates, there were 221 new cases of CM affecting 120 cows; 437 lactations never had a case of CM. Environmental pathogens made up 90% (159/176) of etiologic agents isolated. Change in daily milk production following CM was associated with J5 vaccination, days in milk (DIM) at onset of CM, and herd effect as well as each 2-way interaction between the 3 factors. The adjusted daily milk for 21 d following CM was 7.6 kg greater among J5 vaccinates than controls; however, this protective effect of vaccination waned with increasing DIM at onset of CM. A mixed linear model with autoregressive order 1 [AR(1)] correlation structure estimated the daily milk production of any cow (whether or not she had CM) on a given DIM. Cows with CM caused by nonagalactiae streptococci, Staphylococcus aureus, Escherichia coli, or Klebsiella lost significant daily milk production for the entire lactation relative to nonmastitic cows. Another mixed linear model for only coliform CM cases (E. coli, Klebsiella, and Enterobacter) within the first 50 DIM showed milk loss for 21 d following coliform CM to be significantly less for J5 vaccinates than for controls, by 6 to 15 kg per day. Cows were significantly less likely to become pregnant if they had CM caused by E. coli (42% pregnant) or Streptococcus spp. (38% pregnant), whereas 78% (342/437) of cows with no mastitis conceived. Days open (number of days from calving until pregnancy) averaged 131 d for cows with no CM and 162 d for cows that had at least one case of CM. Days until conception, days until last breeding, days open, times bred, and percentage of cows pregnant by 200 DIM were not changed with J5 vaccination. Nonetheless, an important benefit of the use of J5 bacterin appears to be reduction of the loss of daily milk production following CM, whether all cases or only those caused by coliform bacteria were considered.     

Risk factors for isolation of Staphylococcus aureus or Streptococcus dysgalactiae from milk culture obtained approximately 6 days post calving

Milk culture results at approximately 6 d post calving were assessed in a 2-year retrospective single-cohort study in 178 Norwegian herds. A combined teat dipping and selective antibiotic therapy trial was performed in these herds where cows with composite milk somatic cell count (CMSCC) >100,000 cells/ml before drying-off (geometric mean of the last three CMSCC test-days) and isolation of Staphylococcus aureus or Streptococcus dysgalactiae were selected for either short-acting lactation antibiotic treatment or long-acting dry cow antibiotic treatment. Milk culture results at approximately 6 d post-calving were available from 437 treated cows and 3061 non-treated cows before drying-off and separate multivariable logistic regression models were ran for these two groups. Risk factors associated with isolation of Staph. aureus 6 d post calving for non-treated cows were CMSCC >400,000 cells/ml before drying-off v. <400,000 cells/ml (Odd ratio (OR) = 2.4) and clinical mastitis (CM) in the previous lactation v . non-treated (OR=1.5). Risk factors associated with Staph. aureus 6 d post calving for treated cows was a CMS > 200,000 cells/ml before drying-off v. <200,000 cells/ml (OR=2.3) and CM in the previous lactation verus non-treated (OR=1.7). For non-treated cows it was 1.7 times more likely to isolate Str. dysgalactiae 6 d post-calving if the CMSCC was > 50,000 cells/ml compared with <50,000 cells/ml. For treated cows it was 3.7-5.8-times more likely to isolate Str. dysgalactiae 6 d post calving if given short-acting lactation formula at quarter level compared with long-acting dry cow formula used at cow level. Regular use of iodine post-milking teat disinfection (PMTD) did not influence the isolation of Staph. aureus 6 d post calvin, but it was less likely to isolate Str. dysgalactiae 6 d post calving if iodine PMTD was used regularly rather than irregularly. The external teat sealant had no effect on either of the two bacteria. This study indicates that the CMSCC limit for sampling cows before drying-off can be reduced to 50,000 cells/ml in herds with a Str. dysgalactiae problem. Iodine PMTD should also be recommended in these herds. Cows with a CMSCC > 400,000 cells/ml prior to drying-off should receive long-acting dry cow formula irrespective of the milk result.     

A cohort study of the effect of Streptococcus agalactiae on milk yield and somatic cell count in Norwegian dairy cows

The primary objective of the present study was to estimate the effect of Streptococcus agalactiae intramammary infection on milk production and somatic cell count (SCC) in Norwegian dairy cows. A secondary objective was to assess differences in the effect of common Strep. agalactiae sequence types (ST) found in Norwegian dairy herds. We performed a cohort study combining registry data with sequence-type data from Strep. agalactiae isolates. Herds in which Strep. agalactiae had been detected in individual animals (bacteriological culture or quantitative PCR) between 2012 and 2015 were included. We accessed monthly test-day milk yield records for the entire period to compare milk yield and SCC between cows that were Strep. agalactiae positive and all other cows, within each herd. The study sample consisted of 150 herds, 15,757 cows, 30,850 lactations, and 204,126 test days. We evaluated the effects of Strep. agalactiae on test-day milk yield and SCC using mixed linear regression models, controlling for clustering by herd, cow, and lactation. Multilocus sequence typing of Strep. agalactiae was available for isolates from 86 herds. Additional models were fit to a subset of herds (n = 59) in which ST1, ST23, ST103, and ST196 had been found, to compare the effects of ST on milk production and SCC. In the period 3 to 2 mo before diagnosis, Strep. agalactiae-positive cows produced an average of 1.3 kg more DIM-adjusted milk/d than their negative herd mates. At the time of diagnosis, production was on average 0.13 kg less DIM-adjusted milk/d in Strep. agalactiae-positive cows than in negative cows; 2 to 3 mo after diagnosis, they produced 1.24 kg less DIM-adjusted milk/d than negative cows. Losses persisted for the rest of the investigated period. Cows with ST23, ST103, and ST196 followed a similar pattern as the overall analysis with respect to milk production, whereas ST1-affected cows produced similar amounts of milk before diagnosis as the negative cows. Cows with ST1 experienced the largest milk loss 1 to 2 mo after diagnosis but then recovered to some extent; for cows with ST103, the severe milk loss persisted for the rest of the investigation period. The cow-associated ST103 elicited a lower response in peak SCC compared with ST23, ST103, and ST196. The results indicate an effect of Strep. agalactiae on milk production and SCC. Production was lowest 2 to 3 mo after a positive sample. Peak SCC was reached the month before diagnosis, with notable differences between sequence types.     

Effect of prepartum dry cow antibiotic treatment in dairy heifers on udder health and milk production

A high percentage of heifers calve with intramammary infections. One of the measures available to control intramammary infections is treatment with antibiotics before calving. In this study, the effects of prepartum treatment of nonlactating heifers with a 600-mg cloxacillin dry cow treatment on the prevalence of culture-positive milk samples at calving and 10 to 14 d in milk (DIM), the incidence of clinical mastitis, somatic cell count (SCC), and milk production during first lactation were quantified. A total of 184 heifers on 13 dairy farms were treated with antibiotics 8 to 10 wk before the expected calving date. Another 185 heifers served as untreated controls. Coagulase-negative staphylococci were the most frequently isolated group of bacteria in the treatment and control groups at calving (32 and 42%), and at 10 to 14 DIM (15 and 19%), respectively. The prevalence of minor pathogens at calving was lower in the treatment group compared with the control group (34 and 43%, respectively). Staphylococcus aureus was the most frequently isolated major pathogen in the treated and control heifers at calving (4 and 5%) and at 10 to 14 DIM (2 and 3%), respectively. The prevalence of major pathogens was lower in the treated heifers at 10 to 14 DIM compared with the control group (4 and 6%, respectively). Cumulative incidence risk of clinical mastitis during the lactation was 9 and 18% in the treatment and control groups, respectively. Treatment with cloxacillin 8 to 10 wk before calving resulted in a lower prevalence of culture-positive milk samples at calving and lower quarter milk SCC in early lactation [30,000 ± 4,600 (standard deviation) cells/mL in treated heifers versus 40,000 ± 4,600 cells/mL in control heifers], and was associated with lower average test-day SCC (55,000 ± 1,400 cells/mL in treated heifers versus 71,000 ± 1,500 cells/mL in control heifers) and lower incidence of clinical mastitis throughout lactation. The improved udder health resulted in a higher average test-day milk production in the first lactation (24.5 ± 3.2 kg in treated heifers versus 23.6 ± 3.1 kg in control heifers). Dairy farms with heifer mastitis problems need to analyze their mastitis management. Prepartum treatment of heifers with dry cow antibiotics may be helpful by decreasing the prevalence of mastitis-causing pathogens at calving and at 10 to 14 DIM.     

Effect of milk yield characteristics, breed, and parity on success of the first insemination in Dutch dairy cows

The objective of this study was to determine the contribution of cow factors to the probability of a successful first insemination (SFI). The investigation was performed with 51,791 lactations from 1,396 herds obtained from the Dutch dairy cow database of the Cattle Improvement Co-operative (CRV). Cows that had the first insemination (AI) between 40 and 150 d postpartum were selected. The first AI was classified as successful when cows were not reinseminated and either calved between 267 and 295 d later or were culled within 135 to 295 d after first AI. The lactation curve characteristics of individual lactations were estimated by Wilmink's curve using the test-day milk records from CRV. The lactation curve characteristics (peak milk yield, milk yield at the first-AI date, time of peak yield (PT), and milk persistency) were calculated. Breed, parity, interval from calving to first AI (CFI), lactation curve characteristics, milk production traits, moment of AI related to PT (before or after PT), calf status, month of AI, and month of calving were selected as independent variables for a model with SFI as a dependent variable. A multivariable logistic regression model was used with farm as a random effect. Overall SFI was 44%. The effect of parity on SFI depended on CFI. The first-parity cows had the greatest SFI (0.43) compared with other parities (0.32-0.39) at the same period of CFI before 60 d in milk (DIM), and cows in parity ≥5 had the least SFI (0.38-0.40) when AI was after 60 DIM. After 60 DIM, extending CFI did not improve SFI in the first-parity cows, but SFI was improved in multiparous cows. Holstein-Friesian cows had lesser SFI (0.37) compared with cross-breed cows (0.39-0.46). Twin and stillbirth calving reduced SFI (0.39) compared with a single female calf (0.45) or a male calf (0.43) calving. The SFI in different months of AI varied and depended on CFI. Cows that received AI before 60 DIM had a lesser SFI, especially in March, June, and July (0.18, 0.35, and 0.34, respectively). Artificial insemination before PT reduced SFI (0.39) in comparison with AI after PT (0.44). The effect of milk yield at the first-AI date on SFI varied depending on CFI. After 60 DIM at the same period of CFI, a high level of milk yield at the first-AI date reduced SFI. In conclusion, knowledge of the contribution of cow factors on SFI can be applied to support decision making on the moment of insemination of an individual cow in estrus.     

Clinical outcomes and molecular genotyping of Staphylococcus aureus isolated from milk samples of dairy primiparous Mediterranean buffaloes (Bubalus bubalis)

Staphylococcus aureus is one of the most important pathogens causing mastitis in dairy cows and in Mediterranean buffaloes. Genotype B (GTB) is contagious in dairy cows and may occur in up to 87% of cows of a dairy herd. It was the aim of this study to evaluate genotypes present, clinical outcomes, and prevalence of Staph. aureus in milk samples of primiparous Mediterranean dairy buffaloes. Two hundred composite milk samples originating from 40 primiparous buffaloes were collected from May to June 2012, at d 10, 30, 60, 90, and 150 d in milk (DIM) to perform somatic cell counts and bacteriological cultures. Daily milk yields were recorded. Before parturition until 40 to 50 DIM, all primiparous animals were housed separated from the pluriparous animals. Milking was performed in the same milking parlor, but the primiparous animals were milked first. After 50 DIM, the primiparous were mixed with the pluriparous animals, including the milking procedure. Individual quarter samples were collected from each animal, and aliquots of 1 mL were mixed and used for molecular identification and genotyping of Staph. aureus. The identification of Staph. aureus was performed verifying the presence of nuc gene by nuc gene PCR. All the nuc-positive isolates were subjected to genotype analysis by means of PCR amplification of the 16S-23S rRNA intergenic spacer region and analyzed by a miniaturized electrophoresis system. Of all 200 composite samples, 41 (20.5%) were positive for Staph. aureus, and no genotype other than GTB was identified. The prevalence of samples positive for Staph. aureus was 0% at 10 DIM and increased to a maximum of 22/40 (55%) at 90 DIM. During the period of interest, 14 buffaloes tested positive for Staph. aureus once, 6 were positive twice, and 5 were positive 3 times, whereas 15 animals were negative at every sampling. At 90 and 150 DIM, 7 (17.5%) and 3 buffaloes (7.5%), respectively, showed clinical mastitis (CM), and only 1 (2.5%) showed CM at both samplings. At 60, 90, and 150 DIM, 1 buffalo was found with subclinical mastitis at each sampling. At 30, 60, 90, and 150 DIM, 2.5 (1/40), 22.5 (9/40), 35 (14/40), and 10% (4/40) were considered affected by intramammary infection, respectively. Buffaloes with CM caused by Staph. aureus had statistically significantly higher mean somatic cell count values (6.06 ± 0.29, Log₁₀ cells/mL ± standard deviation) and statistically significantly lower mean daily milk yields (7.15 ± 1.49, liters/animal per day) than healthy animals (4.69 ± 0.23 and 13.87 ± 2.64, respectively), buffaloes with IMI (4.82 ± 0.23 and 11.16 ± 1.80, respectively), or with subclinical mastitis (5.47 ± 0.10 and 10.33 ± 0.68, respectively). Based on our knowledge, this is the first time that Staph. aureus GTB has been identified in milk samples of dairy Mediterranean buffaloes.     

Identification of potential markers in blood for the development of subclinical and clinical mastitis in dairy cattle at parturition and during early lactation

Our objective was to identify specific blood markers as risk factors for the development of mastitis during early lactation. We used a subset of cows from a larger experiment that consisted of a total of 634 lactations from 317 cows. Cows were of 3 breeds and ranged from parity 1 to 4. Blood samples were collected weekly from 56 d before expected calving date through 90 d in milk (DIM). Blood was analyzed for several hormones, metabolites, and enzymes, and energy intake and energy balance were calculated. Veterinary treatment records and daily composite milk somatic cell counts were analyzed and used to determine incidence and severity of mastitis in early lactation. Cows were separated into 2 groups: 1) WK0, consisting of cows that developed clinical mastitis (CM), cows that developed subclinical mastitis (SM), or cows that were healthy (H) during the first 7 DIM; and 2) EL, consisting of CM, SM, or H cows during wk 2 through 13 of lactation. Data were adjusted for numerous fixed effects (e.g., parity, breed, season, and DIM) before statistical analysis. The time of mastitis (TOM) was recorded as the DIM in which the first rise in somatic cell count was observed and was recorded as TOM = 0. The time before and after TOM was distinguished as ± n wk relative to TOM = 0. Healthy cows were paired with either a SM or CM cow and the TOM for each H cow was equal to the TOM for its paired SM or CM cow. Data from wk −1 and −2 relative to TOM were analyzed for group WK0 and EL, respectively. For all parameters, SM cows did not differ from H cows from either group. The CM cows had higher nonesterified fatty acid levels and a tendency toward higher β-hydroxybutyrate levels than H cows before mastitis for both groups. For group WK0, glucose was higher −1 wk relative to calving in CM than H cows. For group EL, aspartate aminotransferase was higher −2 wk relative to mastitis in CM than H cows during 8 to 90 DIM. All other variables were similar among CM, SM, and H cows for both groups. Our results indicate that substances in blood, especially nonesterified fatty acids and aspartate aminotransferase, may be potential markers for the risk of mastitis in early lactation.     

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.     

Responses to metabolic challenges in dairy cows with high or low milk yield during an extended lactation

Responses of dairy cows with high or low milk yield (MY) beyond 450 d in milk (DIM) to 3 metabolic challenges were investigated. Twelve multiparous Holstein-Friesian cows that calved in late winter in a pasture-based system were managed for a 670-d lactation by delaying re-breeding. Cows were selected for either high MY (18.9 ± 1.69 L/cow per d; n = 6) or low MY (12.3 ± 3.85 L/cow per d; n = 6) at 450 DIM. Cows were housed indoors for 2 periods of 12 d at approximately 460 and 580 DIM. Each cow was fed freshly cut pasture (460 DIM) or pasture silage (580 DIM) plus 6.0 kg of DM barley grain daily (approximately 200 MJ of total metabolizable energy/cow per day). At all other times, cows were managed as a single herd and grazed pasture supplemented with cereal grain to an estimated intake of 180 MJ of metabolizable energy/cow per d. Cows were fitted with a jugular catheter during the final week of each experimental period. Over a period of 3 d, each cow underwent an intravenous glucose tolerance test (0.3 g/kg of body weight), an insulin tolerance test (0.12 IU of insulin/kg of body weight), and a 2-dose epinephrine challenge (0.1 and 1.6 µg/kg of body weight). Cows selected for high MY had greater milk and milk solids yields between 450 and 580 DIM than low MY cows (17.3 vs. 10.8 ± 1.49 kg of milk/d and 2.4 vs. 1.5 ± 0.23 kg of milk solids/d). The results indicated that whole body and peripheral tissue responsiveness to insulin may vary between cows of high and low MY. Following the glucose tolerance test, high MY cows had a lower plasma insulin response with a greater glucose area under the curve than low MY cows. Further, high MY cows had slower plasma glucose clearance compared with low MY cows during an insulin tolerance test. The plasma nonesterified fatty acid (NEFA) responses to the IVGTT and the ITT were similar between cows of high and low MY, but the clearance of NEFA from the plasma following both the IVGTT and ITT were slower at 580 compared with 460 DIM. The sensitivity to epinephrine was greater in high MY cows compared with low MY cows as the glucose and NEFA area under the curve and the percentage change in NEFA were greater in high MY after the low dose epinephrine challenge. However, the lipolytic but not the glucose appearance in response to epinephrine was greater in high MY cows than low MY cows. Following the high dose of epinephrine, the glucose response was lower, but the NEFA response was greater in high MY compared with low MY cows. Cows able to sustain greater MY to 580 DIM had a greater propensity for lipid mobilization, possibly enhancing nutrient partitioning to the mammary gland during the late stages of an extended lactation.     

Determinants of success or failure in the elimination of major mastitis pathogens in selective dry cow therapy.

Discriminant factors for the elimination of major pathogens (mainly Staphylococcus aureus and Streptococcus dysgalactiae) were identified from a randomized, double-blind, field study on dry cow therapy. From an original study of 686 cows, 269 fulfilled the criteria for this analysis: 93 from the control group, 83 from group C (treated with Benestermycin vet. "LEO") and 93 from group D (treated with Leocillin with Dihydrostreptomycin vet. "LEO"). A "success" cow was defined as a cow that had all quarters healthy in two tests after dry period. Isolation of a major pathogen (mainly Staph. aureus or Strep. dysgalactiae) in any quarter in both samples after the dry period was defined as "failure." Better elimination was associated with short-acting preparations (therapy D) rather than with long-acting therapy C (odds ratio = 0.32), as was a smaller mean value of the last three composite milk somatic cell counts (CMSCC) before dry cow therapy. Cows with a major pathogen in the right hind quarter at drying off or at least one case of acute clinical mastitis during the previous lactation were more likely to have a major pathogen in the next lactation (odds ratio = 4.1 and 3.6, respectively). Young cows without major pathogens at drying off should not be recommended for dry cow therapy, and cows with high CMSCC should be considered for culling if their profiles also show previous acute clinical mastitis in combination with generally high CMSCC and low cure rates in the herd.     

Temporal patterns of lost milk production following clinical mastitis in a large Michigan Holstein herd

Cows with three hundred and sixteen cases of clinical mastitis were sampled by microbiologic culture during a 6-mo period on a 1700-cow Michigan Holstein dairy farm. Daily milk weights were obtained on all cows before clinical onset and for 60 d after onset. Predicted post-mastitis production, projected on the basis of premastitis production and the lactation curves of contemporary non-mastitic herdmates, was compared with actual daily milk production during the 60 d following clinical onset. Cows experiencing clinical mastitis produced approximately 341 kg less salable milk during the 60 d after clinical onset compared with projected production. This milk loss included both decreased production and milk withheld from market following antibiotic treatment. Pluriparous cows lost 2.06 times as much milk as first lactation cows, and cows with mastitis occurring before 150 d in lactation lost 1.40 times as much milk when compared with other cows. Cows with mastitis occurring in the winter showed a milk loss 1.37 times greater than cows with mastitis in summer. The identity of the mastitis agent isolated from the clinical case was not strongly associated with the drop in milk production in the 60 d following clinical onset.     

Streptococcus dysgalactiae isolates at calving and lactation performance within the same lactation

The objective of the study was to investigate the association between early lactation Streptococcus dysgalactiae isolates and milk yield, somatic cell count (SCC), clinical mastitis, and culling in the same lactation. The 178 commercial dairy herds were randomly placed into 3 penicillin- or penicillin-dihydrostreptomycin-based dry-cow treatments and 3 different postmilking teat disinfection groups—negative control, iodine, or external teat sealant. All cows were sampled in early lactation, and Strep. dysgalactiae-positive and culture-negative cows were followed throughout the remainder of the lactation. Mixed models, including repeated measurements, with test-day observation as dependent variable, were used to compare milk yield, SCC, and available milk quality variables throughout the remaining lactation. Survival analyses, using a positive frailty model to account for any herd random effects, were used to estimate the hazard ratio for clinical mastitis and culling. Streptococcus dysgalactiae-positive cows had a significantly higher SCC throughout the lactation compared to culture-negative cows. For primiparous or multiparous cows, respectively, the differences in the geometric mean SCC between Strep. dysgalactiae-positive and culture-negative cows was 197,000 or 280,000 cells/mL at the beginning of the lactation, 24,000 or 46,000 cells/mL in mid lactation, and 39,000 or 111,000 cells/mL at the end of the lactation. Streptococcus dysgalactiae-positive primiparous or multiparous cows produced 334 or 246 kg less milk, respectively, during a 305-d lactation compared with culture-negative cows. Compared with culture-negative cows, the hazard ratios for clinical mastitis in Strep. dysgalactiae-positive cows were 2.3 (1.9 to 2.9) and 1.6 (1.3 to 2.0) for culling. For cows with both Strep. dysgalactiae and Staphylococcus aureus isolates, the hazard ratio for culling significantly increased to 2.5 (1.9 to 3.2).     

Heifers infected with coagulase-negative staphylococci in early lactation have fewer cases of clinical mastitis and higher milk production in their first lactation than noninfected heifers

Intramammary infections (IMI) in recently calved dairy heifers are more common than was formerly believed but their relevance for future performance has been studied only rarely. In the present study, the association between the IMI status of fresh heifers and their subsequent udder health, milk production, and culling in first lactation was explored. Quarter milk samples were collected between 1 and 4 d in milk (DIM) and between 5 and 8 DIM from 191 dairy heifers in 20 dairy herds for bacteriological culturing and somatic cell count (SCC) analysis. Monthly milk recording data including composite milk SCC and test-day milk yield (MY) were obtained for the first 285 DIM or until culling. Farmer-recorded clinical mastitis cases were available. Data were analyzed using mixed models and survival analysis. Approximately 80% of the fresh heifers (79.8%) had at least one culture-positive quarter. Coagulase-negative staphylococci (CNS) were the most frequently isolated pathogens (72%), followed by esculin-positive streptococci (4.6%) and Staphylococcus aureus (3.5%). Overall geometric mean SCC at quarter level decreased between the first and second samplings from 348,000 to 116,000 cells/mL. Heifers infected with CNS had an intermediate average test-day SCC (84,000 cells/mL) during the first 285 DIM compared with noninfected heifers (53,000 cells/mL) and heifers infected with major pathogens (195,000 cells/mL). Heifers infected with major pathogens had a much lower average daily MY (18.3 kg) during first lactation compared with noninfected animals (21.3 kg). That CNS-infected heifers out-produced their noninfected counterparts could be at least partially explained by their significantly lower incidence of clinical mastitis (incidence risk 3.6 vs. 21.0%) during first lactation compared with noninfected heifers. We conclude that although CNS cause the majority of IMI in heifers around calving, they should not be a reason for serious concern.     

Associations between CXCR1 polymorphisms and pathogen-specific incidence rate of clinical mastitis,test-day somatic cell count,and test-day milk yield

The CXCR1 gene plays an important role in the innate immunity of the bovine mammary gland. Associations between single nucleotide polymorphisms (SNP) CXCR1c.735C>G and c.980A>G and udder health have been identified before in small populations. A fluorescent multiprobe PCR assay was designed specifically and validated to genotype both SNP simultaneously in a reliable and cost-effective manner. In total, 3,106 cows from 50 commercial Flemish dairy herds were genotyped using this assay. Associations between genotype and detailed phenotypic data, including pathogen-specific incidence rate of clinical mastitis (IRCM), test-day somatic cell count, and test-day milk yield (MY) were analyzed. Staphylococcus aureus IRCM tended to associate with SNP c.735C>G. Cows with genotype c.735GG had lower Staph. aureus IRCM compared with cows with genotype c.735CC (rate ratio = 0.35, 95% confidence interval = 0.14–0.90). Additionally, a parity-specific association between Staph. aureus IRCM and SNP c.980A>G was detected. Heifers with genotype c.980GG had a lower Staph. aureus IRCM compared with heifers with genotype c.980AG (rate ratio = 0.15, 95% confidence interval = 0.04–0.56). Differences were less pronounced in multiparous cows. Associations between CXCR1 genotype and somatic cell count were not detected. However, MY was associated with SNP c.735C>G. Cows with genotype c.735GG out-produced cows with genotype c.735CC by 0.8 kg of milk/d. Results provide a basis for further research on the relation between CXCR1 polymorphism and pathogen-specific mastitis resistance and MY.     

Evaluation of test-day milk somatic cell count to predict intramammary infection in late lactation grazing dairy cows

The use of selective dry cow antimicrobial therapy requires precisely differentiating cows with an intramammary infection (IMI) from uninfected cows close to drying-off to enable treatment allocation. Milk somatic cell count (SCC) is an indicator of an inflammatory response in the mammary gland and is usually associated with IMI. However, SCC can also be influenced by cow-level variables such as milk yield, lactation number, and stage of lactation. In recent years, predictive algorithms have been developed to differentiate cows with IMI from cows without IMI based on SCC data. The objective of this observational study was to explore the association between SCC and subclinical IMI, taking cognizance of cow-level predictors on Irish seasonal spring calving, pasture-based systems. Additionally, the optimal test-day SCC cut-point (maximized sensitivity and specificity) for IMI diagnosis was determined. A total of 2,074 cows across 21 spring calving dairy herds with an average monthly milk weighted bulk tank SCC of ≤200,000 cells/mL were enrolled in the study. Quarter-level milk sampling was carried out on all cows in late lactation (interquartile range = 240–261 d in milk) for bacteriological culturing. Bacteriological results were used to define cows with IMI when ≥1 quarter sample resulted in bacterial growth. Cow-level test-day SCC records were provided by the herd owners. The ability of the average, maximum, and last test-day SCC to predict infection were compared using receiver operator curves. Predictive logistic regression models tested included parity (primiparous or multiparous), yield at last test-day, and a standardized count of high SCC test-days. In total, 18.7% of cows were classified as having an IMI, with first-parity cows having a higher proportion of IMI (29.3%) compared with multiparous cows (16.1%). Staphylococcus aureus accounted for the majority of these infections. The last test-day SCC was the best predictor of infection with the highest area under the curve. The inclusions of parity, yield at last test-day, and a standardized count of high SCC test-days as predictors did not significantly improve the ability of last test-day SCC to predict IMI. The cut-point for last test-day SCC that maximized sensitivity and specificity was 64,975 cells/mL. This study indicates that in Irish seasonal pasture-based dairy herds with low bulk tank SCC, the last test-day SCC (interquartile range days in milk = 221–240) is the best predictor of IMI in late lactation.     

Selective dry cow therapy effect on milk yield and somatic cell count: A retrospective cohort study

Antibiotic dry cow therapy (aDCT) at the end of lactation is an effective mastitis control measure. Selective dry cow therapy means that only infected or presumed-infected cows are treated, instead of aDCT being used as a treatment for all cows. Because antibiotic resistance poses a global threat, livestock production is under increasing pressure to reduce antibiotic use. Changes in management should not, however, impair animal welfare or cause significant economic losses. Our objective was to compare milk yield and somatic cell count (SCC) between aDCT-treated and untreated cows in herds that used selective aDCT, taking into account risk factors for reduced yield and high SCC. The information source was 2015 to 2017 Dairy Herd Improvement data, with 4,720 multiparous cows from 172 Finnish dairy farms. The response variables were test-day milk yield (kg/d) and naturally log-transformed composite SCC (×1,000 cells/mL) during the first 154 d in milk (DIM). The statistical tool was a linear mixed-effects model with 2-level random intercepts, cows nested within herds, and a first-order autoregressive [AR(1)] correlation structure. The overall proportion of aDCT-treated cows was 25% (1,176/4,720). Due to the interaction effect, SCC on the last test day prior to dry-off affected postcalving milk yield differently in aDCT-treated cows than in untreated cows. A higher SCC prior to dry-off correlated with a greater daily yield difference after calving between cows treated and untreated. The majority of cows had SCC < 200,000 cells/mL before dry-off, and as SCC before dry-off decreased, difference in yield between aDCT-treated and untreated cows decreased. Postcalving SCC was lower for aDCT-treated cows compared with untreated cows. To illustrate, for cows with an SCC of 200,000 cells/mL before dry-off, compared with untreated cows, aDCT-treated cows produced 0.97 kg/d more milk and, at 45 DIM, had an SCC that was 20,000 cells/mL lower. Higher late-lactation SCC and lactational mastitis treatments were associated with higher postcalving SCC. A dry period lasting more than 30 d was associated with higher yields but not with SCC. Our findings indicate that a missed aDCT treatment for a high-SCC cow has a negative effect on subsequent lactation milk yield and SCC, which emphasizes the importance of accurate selection of cows to be treated.