Insulin regulates human mammosphere development and function.

Assessing the role of lactogenic hormones in human mammary gland development is limited due to issues accessing tissue samples and so development of a human in vitro three-dimensional mammosphere model with functions similar to secretory alveoli in the mammary gland can aid to overcome this shortfall. In this study, a mammosphere model has been characterised using human mammary epithelial cells grown on either mouse extracellular matrix or agarose and showed insulin is essential for formation of mammospheres. Insulin was shown to up-regulate extracellular matrix genes. Microarray analysis of these mammospheres revealed an up-regulation of differentiation, cell-cell junctions, and cytoskeleton organisation functions, suggesting mammosphere formation may be regulated through ILK signalling. Comparison of insulin and IGF-1 effects on mammosphere signalling showed that although IGF-1 could induce spherical structures, the cells did not polarise correctly as shown by the absence of up-regulation of polarisation genes and did not induce the expression of milk protein genes. This study demonstrated a major role for insulin in mammary acinar development for secretory differentiation and function indicating the potential for reduced lactational efficiency in women with obesity and gestational diabetes.

Adverse effects of LPS on membrane proteins in lactating bovine mammary epithelial cells.

Mastitis causes a decrease in milk yield and abnormalities in milk components from dairy cows. Escherichia coli and the E. coli lipopolysaccharide (LPS) cell wall component directly downregulate milk production in bovine mammary epithelial cells (BMECs). However, the detailed mechanism by which this occurs in BMECs remains unclear. Various membrane proteins, such as immune sensors (Toll-like receptors, TLR), nutrient transporters (glucose transporter and aquaporin), and tight junction proteins (claudin and occludin) are involved in the onset of mastitis or milk production in BMECs. In this study, we investigated the influence of LPS on membrane proteins using an in vitro culture model. This mastitis model demonstrated a loss of glucose transporter-1 and aquaporin-3 at lateral membranes and a decrease in milk production in response to LPS treatment. LPS disrupted the tight junction barrier and caused compositional changes in localization of claudin-3 and claudin-4, although tight junctions were maintained to separate the apical membrane domains and the basolateral membrane domains. LPS did not significantly affect the expression level and subcellular localization of epidermal growth factor receptor in lactating BMECs with no detectable changes in MEK1/2-ERK1/2 signaling. In contrast, NFκB was concurrently activated with temporal translocation of TLR-4 in the apical membranes, whereas TLR-2 was not significantly influenced by LPS treatment. These findings indicate the importance of investigating the subcellular localization of membrane proteins to understand the molecular mechanism of LPS in milk production in mastitis.

Niacin stimulates EPH4EV mammary epithelial cell proliferation and mammary gland development in pubertal mice through activation of AKT/mTOR and ERK1/2 signaling pathways.

Previous studies have shown the effects of vitamins on the development of the mammary gland. However, the role of niacin in this process has not been reported. Therefore, the aim of this study is to investigate the effects of niacin on mammary gland development in pubertal mice and to use a mouse mammary epithelial cell line to study the underlying mechanism. The results showed that niacin could activate the AKT/mTOR and ERK signaling pathways and increase phosphorylation of 4EBP1 to promote the synthesis of cell proliferation markers, leading to the dissociation of the Rb-E2F1 complex in mMECs. In addition, 0.5% niacin promoted mammary duct development, increased the expression of cyclin D1/D3 and PCNA and activated Akt/mTOR and ERK1/2 in the mammary glands of pubertal mice. These results strongly suggest that niacin stimulates mammary gland development in pubertal mice through the Akt/mTOR and ERK1/2 signaling pathways.

Mycoplasma bovis subverts autophagy to promote intracellular replication in bovine mammary epithelial cells cultured in vitro.

Mycoplasma species are the smallest prokaryotes capable of self-replication. To investigate Mycoplasma induced autophagy in mammalian cells, Mycoplasma bovis (M. bovis) and bovine mammary epithelial cells (bMEC) were used in an in vitro infection model. Initially, intracellular M. bovis was enclosed within a membrane-like structure in bMEC, as viewed with transmission electron microscopy. In infected bMEC, increased LC3II was verified by Western blotting, RT-PCR and laser confocal microscopy, confirming autophagy at 1, 3 and 6 h post-infection (hpi), with a peak at 6 hpi. However, the M. bovis-induced autophagy flux was subsequently blocked. P62 degradation in infected bMEC was inhibited at 3, 6, 12 and 24 hpi, based on Western blotting and RT-PCR. Beclin1 expression decreased at 12 and 24 hpi. Furthermore, autophagosome maturation was subverted by M. bovis. Autophagosome acidification was inhibited by M. bovis infection, based on detection of mCherry-GFP-LC3 labeled autophagosomes; the decreases in protein levels of Lamp-2a indicate that the lysosomes were impaired by infection. In contrast, activation of autophagy (with rapamycin or HBSS) overcame the M. bovis-induced blockade in phagosome maturation by increasing delivery of M. bovis to the lysosome, with a concurrent decrease in intracellular M. bovis replication. In conclusion, although M. bovis infection induced autophagy in bMEC, the autophagy flux was subsequently impaired by inhibiting autophagosome maturation. Therefore, we conclude that M. bovis subverted autophagy to promote its intracellular replication in bMEC. These findings are the impetus for future studies to further characterize interactions between M. bovis and mammalian host cells.

Longitudinal study on the effects of intramammary infection with non-aureus staphylococci on udder health and milk production in dairy heifers.

We conducted a longitudinal study to evaluate the effect of non-aureus staphylococci (NAS) causing subclinical intramammary infections (IMI) on quarter milk somatic cell count (qSCC) and quarter milk yield (qMY). In total, 324 quarters of 82 Holstein Friesian heifers were followed from calving to 130 d in milk (DIM) and were sampled 10 times each at 14-d intervals. The IMI status of each quarter was determined based on bacterial culture results at the current and previous or next sampling day, or both. The qSCC was determined on each sampling day and the average qMY on sampling day was available through stored daily milk weight data in the management program of the automatic milking system. A transient IMI (tIMI) was defined as a case where a specific pathogen was isolated from a quarter on only one sampling day and not on the previous or next sampling day. When the same bacterial strain, as defined by random amplification of polymorphic DNA-PCR, was isolated from the same quarter on multiple sampling days, it was defined as a persistent IMI (pIMI) status on those sampling days; a pIMI episode was defined as the combination of multiple consecutive pIMI statuses with the same bacterial strain on different sampling days. During this study, 142 subclinical IMI with NAS occurred in 116 different quarters from 64 animals, yielding in total 304 NAS isolates belonging to 17 different species. The prevalence of NAS was highest in the first 4 DIM. Overall, the predominant species was Staphylococcus chromogenes (52% of the isolates), followed by S. epidermidis (9.2%), S. xylosus (8.2%), and S. equorum (5.9%). Staphylococcus chromogenes was the only species for which an effect on qSCC and qMY could be analyzed separately; the other NAS species were considered as a group because of their low prevalence. Eighteen out of 40 IMI (45%) caused by S. chromogenes persisted over at least 2 sampling days, whereas only 10 of 102 (9.8%) IMI caused by other NAS species persisted for at least 2 sampling days. The average duration of pIMI episodes was 110.4 d for S. chromogenes and 70 d for the other NAS species. Remarkably, 17 of the 18 pIMI episodes with S. chromogenes started within the first 18 DIM. The qSCC was highest in quarters having a pIMI with a major pathogen, followed by quarters having a pIMI with S. chromogenes, and a pIMI with other NAS. Transient IMI with other NAS or with a major pathogen caused a small but significantly higher qSCC, whereas the qSCC in quarters having a tIMI with S. chromogenes was not statistically different compared with noninfected quarters. No significant differences in qMY were observed between quarters having a pIMI or tIMI with S. chromogenes or with the other NAS species compared with noninfected quarters, despite the higher qSCC. Quarters having a pIMI with major pathogens showed significantly lower daily milk production. Surprisingly, quarters that cured from an IMI with S. chromogenes had a significantly lower qMY than noninfected quarters.

Got Milk? Identifying and Characterizing Lactation Defects in Genetically-Engineered Mouse Models.

The ability to produce and expel milk is important for the health and survival of all mammals. Nevertheless, our understanding of the molecular events underlying the execution of this process remains incomplete. Whilst impaired mammary gland development and lactational competence remains the subject of focused investigations, defects in these events may also be an unintended consequence of genetic manipulation in rodent models. In this technical report, we outline established and emerging methods to characterize lactation phenotypes in genetically-engineered mouse models. We discuss important considerations of common models, optimized conditions for mating and the importance of litter size and standardization. Methods for quantifying milk production and quality, as well as protocols for wholemount preparation, immunohistochemistry and the preparation of RNA and protein lysates are provided. This review is intended to help guide researchers new to the field of mammary gland biology in the systematic analysis of lactation defects and in the preparation of samples for more focused mechanistic investigations.

The Multi-Faced Role of PAPP-A in Post-Partum Breast Cancer: IGF-Signaling is Only the Beginning.

Insulin-like growth factor (IGF) signaling and control of local bioavailability of free IGF by the IGF binding proteins (IGFBP) are important regulators of both mammary development and breast cancer. A recent genome-wide association study (GWAS) identified small nucleotide polymorphisms that reduce the expression of IGFBP-5 as a risk factor of developing breast cancer. This observation suggests that genetic alterations leading to a decreased level of IGFBP-5 may also contribute to breast cancer. In the current review, we focus on Pregnancy-Associated Plasma Protein A (PAPP-A), a protease involved in the degradation of IGFBP-5. PAPP-A is overexpressed in the majority of breast cancers but its role in cancer has only begun to be explored. More specifically, this review aims at highlighting the role of post-partum involution in the oncogenic function of PAPP-A. Notably, we summarize recent studies indicating that PAPP-A plays a role not only in the degradation of IGFBP-5 but also in the deposition of collagen and activation of the collagen receptor discoidin 2 (DDR2) during post-partum involution. Finally, considering the immunosuppressive microenvironment of post-partum involution, we also discuss the unexpected finding made in Ewing Sarcoma that PAPP-A plays a role in immune evasion. While the immunosuppressive role of PAPP-A in breast cancer remains to be determined, collectively these studies highlight the multifaced role of PAPP-A in cancer that extends well beyond its effect on IGF-signaling.

Genetic parameters of weeping teats in Italian Saanen and Alpine dairy goats and their relationship with milk production and somatic cell score.

This paper reports a quantitative genetics analysis of weeping teats (WT), an abnormality of the mammary gland in goats. Weeping teats are characterized by milk oozing out of the teat or by the presence of multiple cysts near its base. This abnormality has been routinely recorded in Italian Alpine and Saanen goats since 2000 using a score of 0 or 1 (0 = defect not present, 1 = defect present). No information is available on the genetic background of WT or its relationship with production or other udder-related traits. Data obtained by the Italian National Sheep and Goat Breeders Association (Rome, Italy) were used to estimate both heritability of WT and its genetic correlation with milk yield, somatic cell score, and udder traits. The final data set used in the analysis included 2,178 Saanen and 2,309 Alpine primiparous goats kidding from 2009 to 2014. The pedigree data included 7,333 Saanen and 7,421 Alpines, respectively. A threshold-linear multivariate animal model was used to estimate variance and covariance components. A genealogical data analysis was also implemented, including genealogical data completeness, inbreeding, and identification of possible most recent common ancestors. On average, around 4 and 13% of primiparous Saanen and Alpine females kidding from 2009 to 2014 showed mammary gland abnormality, respectively. Weeping teats heritability was 0.27 and 0.26 for Saanen and Alpine, respectively. Genetic correlations between milk production or somatic cell score ranged from -0.16 in Saanen to 0.43 in Alpine, but the standard error of the estimates was very large. Positive genetic correlations were observed among WT and teat characteristics in both Saanen and Alpine. The average inbreeding of abnormality carriers was 2.4 and 5.1 for Saanen and Alpine, respectively. The genealogical data analysis identified 4 common ancestors of affected does in Saanen and 2 in Alpine. These results indicate that WT have a possible genetic background. A genome-wide association study might help in understanding the polygenic or monogenic determination of this abnormality.

Incidence of milk leakage after dry-off in European dairy herds, related risk factors, and its role in new intramammary infections.

The incidence of milk leakage (ML) after dry-off (DO) and related risk factors was studied in 1,175 dairy cows from 41 commercial herds in 8 European countries: Belgium, Czech Republic, Denmark, France, Germany, Italy, the Netherlands, and Spain. Milk leakage was assessed twice for 30 s each during 3 visits at 20 to 24 h, 30 to 34 h, and 48 to 52 h after DO. Information related to dry-cow management and udder health was collected at herd and cow level, including individual somatic cell count (ISCC) from test-day controls and occurrence of clinical mastitis cases from DO until 30 d in lactation. Mixed-effect logistic regression analyses were used to identify possible risk factors for ML and to study the association between ML and new intramammary infections. Intramammary infections were defined as clinical mastitis cases during the dry period and in the first 30 d in lactation or a rise in ISCC from before to after the dry period (threshold: 200,000 cells/mL) or both. Milk leakage was observed in 24.5% of the cows between 20 and 52 h after DO, where the herd incidence varied between 0.0 and 77.8%. The reduction in number of milkings in the weeks before DO had statistically significant effect on the ML incidence. When the milking frequency was reduced from 3 times/d to 2 or maintained at twice a day, cows had 11 (95% CI = 3.43-35.46) or 9 (95% CI = 1.85-48.22) times higher odds of leaking milk, respectively, compared with cows where the milking frequency was reduced from twice to once a day. Also, the milk production 24 h before DO was associated with ML incidence. Hence, cows with a milk production between 13 and 21 L or above 21 L had 2.3 (95% CI = 1.48-3.53) and 3.1 (95% CI = 1.79-5.3) times higher odds of leaking milk, respectively, compared with cows with a milk production below 13 L. A higher ML incidence was present in the group of cows with an average ISCC in the last 3 mo before DO ≥200,000 cells/mL (odds ratio = 1.7; 95% CI = 1.13-2.41) compared with cows with an average ISCC <100,000 cells/mL. Quarters with ML tended to have 2.0 times higher odds of developing clinical mastitis compared with quarters not leaking milk. Cows with ML tended to have 1.5 times higher odds of intramammary infections (i.e., an increase of ISCC or clinical mastitis) compared with cows without ML.

Intramammary antimicrobial treatment of subclinical mastitis and cow performance later in lactation.

The aim of this study was to evaluate long-term therapeutic effects of antimicrobial treatment of recently acquired subclinical mastitis (RASCM) during lactation. Quarter-level clinical mastitis (CM) follow-up, composite somatic cell counts (SCC), and cow-level milk yield later in lactation were evaluated using follow-up data from 2 previously published linked randomized field trials. The first trial randomly assigned antimicrobial treatment with any intramammary product or negative control to culture-positive quarters of cows having a first elevated composite SCC after 2 consecutive low composite SCC measurements. Untreated cows that had a second elevated composite SCC at the next measurement and were staphylococci-positive (i.e., Staphylococcus aureus or non-aureus staphylococci) were randomly assigned to treatment or control. Quarter-level CM cases were reported by the participating herd personnel, and milk yield and composite SCC data were obtained from the regular test-day recording. Frailty survival models were used to evaluate the long-term therapeutic effects of antimicrobial treatment of RASCM on quarter-level CM follow-up. Mixed linear regression models were applied to quantify the effect on milk yield and composite SCC. Data of 638 quarters from 486 cows in 38 herds were available for statistical analyses, of which 229 quarters of 175 cows received antimicrobial treatment for RASCM. Antimicrobial treatment culminated in reduced composite SCC levels later in lactation but did not result in different milk yield levels or CM follow-up compared with control cows. Antimicrobial treatment of cows with RASCM should therefore only be considered in exceptional situations given the current focus on antimicrobial usage reduction in animal husbandry.

Randomized, controlled, superiority study of extended duration of therapy with an intramammary antibiotic for treatment of clinical mastitis.

The objectives of this study were to evaluate the efficacy of therapy of clinical mastitis following intramammary infusion of 2 durations of intramammary antibiotic therapy and to analyze clinical and microbiology data at enrollment or within 4 d of commencement of treatment as potential predictors of subsequent clinical and bacteriological cure. A total of 304 glands were randomly assigned to intramammary treatment with a combination of amoxicillin, clavulanic acid, and prednisolone either 3 times at 12-h intervals (3×12, n = 156) or 5 times at 12-h intervals (5×12 hourly, n = 148). At enrollment (d 0), assessments were made of the cow's demeanor, rectal temperature, teat-end hyperkeratosis, presence of heat or swelling of the gland, presence of clots in the milk, and California Mastitis Test (CMT) score. Cows were revisited on d 4, 14, and 21 and assessed for rectal temperature, the presence of clots in milk, heat or swelling of the gland, and CMT score. Milk samples were collected from enrolled glands for bacteriology (d 0, 4, 14, and 21) and for somatic cell count (d 14 and 21). Data were analyzed using generalized linear models. Treatment group was the key independent variable, but herd was included in the models. Other potentially confounding variables were included where associated at the bivariate level. A second series of analyses were undertaken to predict likelihood of clinical and bacteriological cure using the clinical signs cow age and days in milk, as well as bacteriological results at d 0 and 4. Although we found a higher clinical failure rate in the 3×12 than 5×12 hourly treatment group (28.2 ± 5.0 vs. 13.4 ± 3.6%, respectively), we noted no difference in bacteriological cure percentage (73.3 ± 7.8 vs. 72.0 ± 7.4%, respectively). The presence of heat or swelling at d 4, days in milk at initiation of treatment, age of the cow, and change in CMT score between d 0 and 4 were predictive of clinical and bacteriological cure. We concluded that increasing the duration of treatment resulted in significantly fewer clinical failures, but had no effect on cure proportion, somatic cell count, or new infection rate. Knowledge of cow age and days in milk and clinical signs were predictive of clinical and bacteriological cure proportion and may provide guidance for producers, under veterinary direction, as to when to extend or recommence therapy.

Infection dynamics across the dry period using Dairy Herd Improvement somatic cell count data and its effect on cow performance in the subsequent lactation.

In this study, we studied infection dynamics across the dry period using test-day somatic cell count (SCC) data from 739 Holstein cows from 33 randomly selected commercial dairy herds in Flanders, all of which applied blanket dry-cow therapy at dry-off. First, we determined infection dynamics, combining the last test-day SCC before dry-off and the first test-day SCC after calving. Next, we determined the effect of dry period infection dynamics, adjusting for the level of the second test-day SCC after calving, on the evolution of test-day SCC and milk yield (MY) and on clinical mastitis and culling hazard in the subsequent lactation. Using an SCC threshold of 200,000 cells/mL, 12.6% of the cows considered healthy before dry-off acquired a new intramammary infection (IMI) across the dry period, whereas 66.9% of the cows considered infected before dry-off cured from IMI. Infection dynamics across the dry period significantly affect a cow's SCC, clinical mastitis risk, and culling hazard in the subsequent lactation. Cows with a new IMI, a cured IMI, or a chronic IMI across the dry period had higher test-day SCC than healthy cows, and their test-day SCC evolved differently over time. This was not the case for test-day milk yield, for which no association with infection dynamics was detected. Furthermore, cows with a second test-day SCC <200,000 cells/mL had a lower test-day SCC in the remainder of the lactation than cows with a second test-day SCC ≥200,000 cells/mL, but this association was modified by infection dynamics across the dry period. The lowest test-day SCC in the remainder of the lactation was observed for cows that remained healthy across the dry period combined with a low (<200,000 cells/mL) second test-day SCC. Cows that cured from an IMI present at dry-off and cows with a chronic IMI across the dry period were more likely to develop clinical mastitis (hazard ratio = 2.22 and 2.89; 95% confidence interval = 1.45-3.43 and 1.60-5.20, respectively), and chronic IMI cows were more likely to be culled (hazard ratio = 3.68; 95% confidence interval = 1.64-8.20) in the subsequent lactation compared with healthy cows. This was not true for cows that became infected across the dry period. This study underlines the importance of good udder health management during lactation to prevent IMI at dry-off rather than curing infected cows during the dry period to ensure optimal udder health in the subsequent lactation.

Effect of intramammary infection with non-aureus staphylococci in early lactation in dairy heifers on quarter somatic cell count and quarter milk yield during the first 4 months of lactation.

A longitudinal study was conducted to assess to what extent intramammary infection (IMI) with non-aureus staphylococci (NAS) within the first 4 d after calving in dairy heifers affects quarter milk yield (qMY) and quarter milk somatic cell count (qSCC) during the first 4 mo of lactation. In total, 324 quarters from 82 Holstein Friesian heifers from 3 commercial dairy herds equipped with an automatic milking system were included and followed from calving up to 4 mo in lactation. The automatic milking system allowed us to precisely determine the daily qMY. A milk sample from each quarter was collected in early lactation (between 1 and 4 d in milk) for bacteriological culturing and measurement of the qSCC. Subsequently, milk samples were taken on a biweekly basis for measurement of the qSCC. The milk prolactin level in early lactation was measured, and the relation with NAS IMI was determined. Overall, NAS IMI in early lactation caused only a slight but significant increase in qSCC compared with milk from noninfected quarters during the first 4 mo in lactation, whereas no significant difference in daily qMY was present between NAS-infected and noninfected quarters. The milk prolactin level in early lactation did not differ between NAS-infected and noninfected quarters either. Our data suggest that IMI with NAS (as a group) present shortly after calving do not have an adverse effect on later production. The milk prolactin concentrations were not dissimilar between NAS-infected and noninfected quarters and thus cannot explain why NAS-infected quarters do not produce less than noninfected quarters.

Efficacy of cabergoline in a double-blind randomized clinical trial on milk leakage reduction at drying-off and new intramammary infections across the dry period and postcalving.

The abrupt cessation of milking at dry-off may induce milk leakage, which may increase the risk of new intramammary infections (IMI). This study assessed the efficacy of 1 i.m. injection of 5.6 mg of cabergoline (Velactis, Ceva Santé Animale, Libourne, France) at drying-off on milk leakage after dry-off and new IMI across the dry period and postcalving compared with a placebo (negative control) and an intramammary antibiotic treatment (positive control) under field conditions. The study was a double-blind, randomized, 3-arm, multicenter, clinical trial performed under Good Clinical Practice conditions. Data from 900 dairy cows of various breeds from 63 farms in France, Germany, and Hungary were analyzed. Only quarters with no bacterial growth at drying-off and a cow somatic cell count ≤200,000 cells/mL were included. Quarters infected with major or minor pathogens or cows with high somatic cell count at time of inclusion were excluded. Cows that qualified for the study were visited 7 times in total before and after drying-off and after calving. Presence (yes/no) of milk leakage was recorded on the day after dry-off. A new infected quarter (new IMI) was defined as one with a major pathogen present in any one of the 2 postcalving samples. Two mixed logistic regression models were fitted to the data to evaluate the efficacy of cabergoline in the reduction of milk leakage and new IMI. One i.m. injection of cabergoline at drying-off significantly reduced the incidence of milk leakage the day after dry-off compared with both placebo and antibiotic treatment. Cabergoline-treated cows significantly reduced the risk of new IMI by major pathogens across the dry period and postcalving by 21% when compared with placebo cows (20.5 vs. 26.0%, respectively). However, when milk leakage was added to the model, the significance of cabergoline was reduced. We interpreted this to show that milk leakage is an intervening variable between treatment with cabergoline and lower risk of new IMI. The antibiotic treatment significantly decreased the odds of new IMI compared with both cabergoline and placebo. However, because several countries are currently disallowing the preventive use of antibiotics at dry-off in noninfected quarters, the dry-off facilitator cabergoline may therefore be of particular value to reduce the risk of new IMI across the dry period.

The effect of mastitis management input and implementation of mastitis management on udder health, milk quality, and antimicrobial consumption in dairy herds.

The main objective of this study was to evaluate evolutions in herd-level antimicrobial consumption (AMC) and in udder health and milk quality parameters between herds that received mastitis management input on a regular basis (actively advised by the first author; referred to as intervention herds) and herds that did not (referred to as control herds). Strikingly, herds in the intervention group had a significantly higher prevalence of new intramammary infections compared with those in the control group. No significant differences were observed in the percentage of chronically infected cows, the bulk milk somatic cell count, and the bacterial and coliform count between the intervention and control herds, nor did the herd-level AMC differ between them. Furthermore, the level of mastitis management applied in each herd was assessed and scored [mastitis management score (MMS); higher is better], as was the level of implementation of different recommended mastitis management practices over time, expressed as the mastitis management implementation score (MMIS; higher is better). A large variation was observed in MMS and MMIS in the intervention herds (median = 16 and range = 12 to 22; median = 13 and range = -5 to 31, respectively) and the control herds (median = 16 and range = 9 to 22; median = 9 and range = -13 to 22, respectively). Also, intervention herds in which the herd veterinarian attended each herd visit executed by the first author had a higher MMS and MMIS (median = 20 and 24, respectively) compared with herds in which the veterinarian sometimes (median = 16 and 17, respectively) or never (median = 16.5 and 7.5, respectively) attended the herd visits. Further, the association between MMS or MMIS on one hand and udder health, milk quality, and the herd-level AMC over time on the other was studied using the data of both groups of herds. Better mastitis management was associated with a reduction in the consumption of antimicrobials that are critically important for human health over time and with lower bacterial counts and bulk milk somatic cell count. Better mastitis management can be helpful in obtaining better milk quality and more responsible use of critically important antimicrobials on dairy farms.

Investigating associations between lamb survival to weaning and dam udder and teat scores.

Aims: To examine a range of udder and teat traits in Romney ewes and to describe the frequency with which different scores occur, and to investigate associations between lamb survival to weaning and ewe udder and teat scores. Methods: Mixed-age, mature Romney ewes (n = 1,009) were enrolled from a commercial sheep flock located in the Wellington region of New Zealand in January 2017. A range of udder and teat traits were scored in all ewes, using visual assessment and palpation, at pre-mating (February), pre-lambing (October), docking (November) and weaning (January 2018). During the lambing period each newborn lamb was matched to its dam, with lamb mortalities recorded until weaning. Associations between udder and teat scores and lamb survival to weaning were examined using multivariable models for each udder-scoring time. Results: Records from 981 ewes and 1,822 live-born lambs were included in analyses, with 252 (13.8%) lambs recorded dead between birth and weaning. Lambs born to ewes with pre-mating udder scores of lump or hard had 4.9 (95% CI = 2.6-9.6, p = 0.003) and 3.0 (95% CI = 1.5-6.1, p < 0.001) increased odds of failure to survive to weaning, respectively, compared with lambs whose dams had normal udder scores. Lambs born to ewes with mastitis at docking or weaning had 3.0 (95% CI = 1.5-5.9, p = 0.001) and 3.9 (95% CI = 1.3-11.6, p = 0.013) increased odds of failure to survive to weaning, respectively, compared with lambs whose dams did not have mastitis. Offspring of dams with asymmetrical udders at docking or weaning had 3.3 (95% CI = 2.2-4.9, p < 0.001) and 2.5 (95% CI = 1.5-4.0, p < 0.001) increased odds of failure to survive, respectively, compared with lambs whose dams had symmetrical udders. Conclusion and Clinical Relevance: Pre-mating udder palpation scores of hard or lump were associated with increased odds of lambs not surviving to weaning compared with normal scores, and could be used to identify ewes that are likely to be unsuitable for retaining in the breeding flock. Farmers could also use clinical mastitis scores and udder symmetry scores at docking or weaning to identify ewes whose lambs had greater odds of failure to survive to weaning. However these scores do not provide an indication of future performance, therefore further investigation into the impact of the present season's score on future seasons' lamb survival is required.

Associations between lamb growth to weaning and dam udder and teat scores.

Aims: To investigate associations between lamb growth to weaning and dam udder and teat scores measured at pre-mating, pre-lambing, docking and weaning. Methods: Mature Romney ewes (n = 1,009) were enrolled from a commercial sheep flock located near Masterton, in the Wellington region of New Zealand in 2017. A range of udder and teat traits were scored in all ewes, using visual assessment and palpation, at pre-mating, pre-lambing, docking and weaning. During the lambing period, each newborn lamb was matched to its dam and lamb sex, birthweight and birth-rank were recorded. A rearing rank was allocated to each live-lamb at weaning, when all lambs were weighed (n = 1,570), allowing calculation of daily growth rates (g/day). Associations between udder and teat scores and lamb growth rates to weaning were examined using multivariable models for each udder-scoring time. Results: Growth rates of lambs whose dams had udder palpation scores of hard, or both teats recorded as abnormal, pre-mating were lower than lambs whose dams had normal scores (229.9 (95% CI = 213.2-246.6) vs. 254.5 (95% CI = 245.6-263.5) g/day; p = 0.011) and (227.4 (95% CI = 208.3-246.6) vs. 247.9 (95% CI = 235.7-260.2) g/day; p = 0.024), respectively. Growth rates of lambs whose dams had clinical mastitis at docking or weaning were lower than those without mastitis (215.8 (95% CI = 199.9-231.7) vs. 235.4 (95% CI = 225.4-255.0) g/day; p = 0.007) and (220.0 (95% CI = 205.2-234.8) vs. 254.7 (95% CI = 248.9-260.5) g/day; p < 0.001), respectively. Growth rates of lambs whose dams had asymmetrical udders at docking or weaning were lower than lambs whose dams had symmetrical udders (204.6 (95% CI = 189.7-219.5) vs. 240.2 (95% CI = 225.4-255.0) g/day; p < 0.001) and (223.3 (95% CI = 213.9-232.7) vs. 242.2 (95% CI = 229.4-255.0) g/day; p = 0.014), respectively. Conclusions and Clinical Relevance: Pre-mating udder palpation and teat palpation scores can be used to identify ewes whose lambs are predicted to have lower growth to weaning. Assuming a mean lamb age at weaning of 84.4 days, lambs born to ewes with a pre-mating score of hard would be expected to have a mean weaning weight that was 2.1 kg less than those whose dams had normal scores. Udder palpation, udder symmetry and clinical mastitis scores during lactation were also associated with lamb growth rates. Abbreviation: CALW: Conceptus-adjusted liveweight.

Invited review: Effectiveness of precalving treatment on postcalving udder health in nulliparous dairy heifers: A systematic review and meta-analysis.

Clinical mastitis affects 3% of primiparous dairy cattle (heifers) in the first month after calving. Additionally, the prevalence of intramammary infection (IMI) in the months before first calving is high, resulting in a high prevalence of heifers calving with IMI. Precalving therapy is an accepted recommendation for reducing mastitis in multiparous cows, but prophylactic treatment for heifers is uncommon in North America. Objectives of this study were to (1) quantify changes in postcalving udder health in heifers following application of a precalving treatment; (2) compare effectiveness among various types of treatments; and (3) compare effectiveness of various types of treatments against specific pathogens. A systematic review was conducted comparing interventions aimed at improving udder health in heifers. Of 62 included studies, 48 clinical trials were used in a meta-analysis. Data were synthesized using a random effects model for meta-analysis, followed by sub-group analyses comparing treatment types, and specific pathogens with statistical testing using meta-regression. Occurrence of mastitis (defined as elevated somatic cell count, clinical mastitis, and IMI) was reduced in treated heifers compared with untreated controls with a pooled risk ratio of treated to untreated heifers of 0.56 (95% confidence interval: 0.47 to 0.67). Upon stratification by treatment types, teat sealants and combination therapies (vaccines and antimicrobials; antimicrobials and teat sealants; and all 3) were most effective at improving udder health with pooled risk ratios of 0.40 (95% confidence interval: 0.30 to 0.52) and 0.34 (95% confidence interval: 0.25 to 0.45), respectively. Antimicrobials and vaccines also reduced occurrence of IMI and subclinical and clinical mastitis when compared with untreated heifers. Although variation was observed in the pathogen-specific effectiveness of treatments at reducing rates of disease, antimicrobials, teat sealants, and combinations of vaccines or teat sealants with antimicrobials were consistently effective, whereas vaccines were only effective for contagious pathogens. Recommendations for use of antibiotics should consider their relative benefit while also considering potential for increasing antimicrobial resistance.

Phenotypic and genetic relationships between indicators of the mammary gland health status and milk composition, coagulation, and curd firming in dairy sheep.

The present study investigated the effect of somatic cell count, lactose, and pH on sheep milk composition, coagulation properties (MCP), and curd firming (CF) parameters. Individual milk samples were collected from 1,114 Sarda ewes reared in 23 farms. Milk composition, somatic cell count, single point MCP (rennet coagulation time, RCT; curd firming time, k20; and curd firmness, a30, a45, and a60), and CF model parameters were achieved. Phenotypic traits were statistically analyzed using a mixed model to estimate the effects of the different levels of milk somatic cell score (SCS), lactose, and pH, respectively. Additive genetic, herd, and residual correlations among these 3 traits, and with milk composition, MCP and CF parameters, were inferred using a Bayesian approach. From a phenotypic point of view, higher SCS levels caused a delayed gelification of milk. Lactose concentration and pH were significant for many milk quality traits, with a very intense effect on both coagulation times and curd firming. These traits (RCT, RCT estimated using the curd firming over time equation, and k20) showed an unfavorable increase of about 20% from the highest to the lowest level of lactose. Milk samples with pH values lower than 6.56 versus higher than 6.78 were characterized by an increase of RCT (from 6.00 to 14.3 min) and k20 (from 1.65 to 2.65 min) and a decrease of all the 3 curd firmness traits. From a genetic point of view, the marginal posterior distribution of heritability estimates evidenced a large and exploitable variability for all 3 phenotypes. The mean intra-farm heritability estimates were 0.173 for SCS, 0.418 for lactose content, and 0.206 for pH. Lactose (favorably), and SCS and pH (unfavorably), at phenotypic and genetic levels, were correlated mainly with RCT and RCT estimated using the curd firming over time equation and scarcely with the other curd firming traits. The SCS, lactose, and pH were significantly correlated with each other's. In conclusion, results reported in the present study suggest that SCS, pH, and lactose affect, contemporarily and independently, milk quality and MCP. These phenotypes, easily available during milk recording schemes measured by infrared spectra prediction, could be used as potential indicators traits for improving cheese-making ability of ovine milk.

Udder health of dairy cows fed different dietary energy levels after a short or no dry period without use of dry cow antibiotics.

Reports on the effects of length of dry period (DP) on udder health of cows that were not treated with dry cow antibiotics are scarce. Additionally, the effects of a reduced dietary energy level for cows with a 0-d DP on udder health have not yet been studied. The aims of this study were (1) to compare effects of a 0-d or 30-d DP without use of dry cow antibiotics on udder health across the DP and subsequent lactation in dairy cows fed different dietary energy levels and (2) to evaluate associations between udder health and metabolic status of dairy cows. Five weeks before the expected calving date, Holstein-Friesian dairy cows (n = 115) were blocked for parity, expected calving date, and milk yield and somatic cell count (SCC) at their 2 last test days and were randomly assigned to 2 DP lengths: 0-d DP (n = 77) or 30-d DP (n = 38). Quarter milk samples were taken in wk 5 prepartum and in wk 1 and 5 postpartum. Proportion of quarters with elevated SCC (SCC ≥200,000 cells/mL) and proportion of udder pathogens in quarter milk samples did not differ between DP lengths among weeks. After calving, 102 of these cows were randomly assigned to 3 treatments: a 30-d DP with a standard energy level required for expected milk yield (30-d DP SEL; n = 36), a 0-d DP with the same energy level as cows with a 30-d DP (0-d DP SEL; n = 33), and a 0-d DP with a low energy level (0-d DP LEL, n = 33). From wk 8 of lactation onward, cows received either a glucogenic ration consisting of corn silage and grass silage or a lipogenic ration consisting of grass silage and sugar beet pulp at a standard or low energy level. During wk 1 to 7 postpartum, treatment did not affect SCC or SCC corrected for milk yield. During wk 8 to 44 of lactation, 0-d DP SEL cows had a greater SCC than 0-d DP LEL or 30-d DP SEL cows and had a greater SCC corrected for milk yield than 0-d DP LEL cows. During wk 1 to 44 of lactation, occurrence of at least 1 elevation of SCC (SCC ≥200,000 cells/mL after 2 wk of SCC <200,000 cells/mL) was not different among treatments. The 0-d DP SEL cows but not the 0-d DP LEL cows tended to have a 2.17 times greater hazard of having a case of clinical mastitis at any time in lactation than 30-d DP SEL cows. In wk 1 to 44 of lactation, lower fat- and protein- corrected milk yield and energy intake, greater energy balance, and greater plasma insulin concentration were associated with greater SCC. In conclusion, DP length did not affect udder health in the DP and in early lactation but seemed to decrease udder health for 0-d DP SEL cows in later lactation compared with 30-d DP SEL or 0-d DP LEL cows.