Effects of heat stress abatement on systemic and mammary inflammation in lactating dairy cows.

To examine the effects of evaporative cooling on systemic and mammary inflammation of lactating dairy cows, 30 multiparous Holstein cows (parity = 2.4, 156 d in milk) were randomly assigned to 1 of 2 treatments: cooling (CL) with fans and misters or not (NC). The experiment was divided into a 10-d baseline when all cows were cooled, followed by a 36-d environmental challenge when cooling was terminated for NC cows. The onset of environmental challenge was considered as d 1. Temperature-humidity index averaged 78.4 during the environmental challenge. Milk yield and dry matter intake (DMI) were recorded daily. Blood and milk samples were collected from a subset of cows (n = 9/treatment) on d -3, 1, 3, 7, 14, and 28 of the experiment to measure cortisol, interleukin 10 (IL10), tumor necrosis factor-α (TNF-α), haptoglobin, and lipopolysaccharide binding protein (LBP). Mammary biopsies were collected from a second subset of cows (n = 6/treatment) on d -9, 2, 10, and 36 to analyze gene expression of cytokines and haptoglobin. A subset of cows (n = 7/treatment) who were not subjected to mammary biopsy collection received a bolus of lipopolysaccharides (LPS) in the left rear quarter on d 30 of the experiment. Blood was sampled from cows and milk samples from the LPS-infused quarter were collected at -4, 0, 3, 6, 12, 24, 48, and 96 h relative to infusion, for analyses of inflammatory products. Deprivation of cooling decreased milk yield and DMI. Compared with CL cows, plasma cortisol concentration of NC cows was higher on d 1 but lower on d 28 of the experiment (cooling × time). Deprivation of cooling did not affect circulating TNF-α, IL10, haptoglobin, or LBP. Compared with CL cows, NC cows tended to have higher milk IL10 concentrations but did not show effects in TNF-α, haptoglobin, or LBP. No differences were observed in mammary tissue gene expression of TNF-α, IL10, and haptoglobin. Milk yield declined after LPS infusion but was not affected by treatment. Compared with CL cows, NC cows had greater milk somatic cell count following intramammary LPS infusion. Non-cooled cows had lower circulating TNF-α and IL10 concentrations and tended to have lower circulating haptoglobin concentrations than CL cows. Milk IL10 and TNF-⍺ concentrations were higher 3 h after LPS infusion for NC cows compared with CL cows. Additionally, NC cows tended to have higher milk haptoglobin concentration after LPS infusion than CL cows. In conclusion, deprivation of evaporative cooling had minimal effects on lactating cows' basal inflammatory status, but upregulated mammary inflammatory responses after intramammary LPS infusion.

Late embryo mortality in Holstein cows inseminated with Holstein or Limousine frozen semen.

The objective of this study was to compare the late embryo mortality (LEM) rate (losses approximately between 32 and 53 days of gestation) and Pregnancy Specific Protein B (PSPB) and progesterone (P4) concentrations on day 32 post AI in Holstein cows bred with either Holstein or Limousine semen. A sample size of 1082 cows per group diagnosed pregnant between 28- and 35-days post breeding was calculated. The study consisted of evaluating LEM (%) in a cohort of Holstein cows bred with Holstein semen (HO × HO) or Limousine semen (HO × LM), to compare pregnancy loss from 28 to 35 days post breeding to 50-57 days post breeding. A logistic regression model to compare embryo losses was developed considering as main explanatory variable the cohort (HO × HO embryo vs. HO × LM embryo), correcting by lactation number, breeding season, days to breeding and AI technician. HO × HO embryos had greater LEM (15.16%) than HO × LM embryos (9.79%). Cows bred in summertime had higher LEM (15.23%) than cows bred in no-summertime (9.88%). There were no differences among AI technicians. Within summertime there was no difference in LEM (%) between groups within each lactation number; yet, within no-summertime, LEM (%) was higher in HO × HO than HO × LM within each lactation number. Pregnancy SPB optical densities were significantly greater in the HO × HO than in the HO × LM (p = .023) group; yet, the concentration of P4 was not different between groups (p > .05).

Effect of a very low negative dietary cation-anion difference (DCAD) diet on plasma and urine metabolomics of prepartum Holstein cows.

The objectives of this cross-sectional, nonintervention, observational study were to compare urine and blood parameters between cows consuming a positive dietary cation-anion difference (DCAD) diet [early dry cows, DCAD + 250 mEq/kg of dry matter (DM), n = 15] with the same cows consuming a negative DCAD diet (-220 mEq/kg of DM) 10 d after moving them from the early dry to the prepartum group. The most remarkable finding was that cows consuming the anionic diet had very low urine pH and very low base excess in blood, suggestive of uncompensated metabolic acidosis. Importantly, the metabolomics data revealed that only urine concentrations of essential and aromatic amino acids were decreased, and that concentrations of total nonessential amino acids and glucogenic amino acids were increased in plasma and reciprocally decreased in urine, suggesting that the cows fed anionic salts were attempting to meet a high glucose demand by mobilizing gluconeogenic amino acid reserves. Notably, the dietary anionic salts exerted marked effects on glycerophospholipids, with a reduction in most phosphatidylcholine containing diacyl (PC aa) and acyl-alkyl (PC ae) moieties in plasma and urine. Further characterization of these metabolomic profiles may lead to the development of novel biomarkers to identify cows susceptible to metabolic acidosis and other metabolic diseases.

Effect of fish oil and canola oil supplementation on immunological parameters, feed intake, and growth of Holstein calves.

Supplemental n-3 fatty acids (FA) may support better immune responses than n-6 and n-9 FA in dairy calves. The objective was to evaluate the effect of n-3 FA, supplemented as a fish oil product (FO) in the milk replacer (MR), in comparison to n-6 and n-9 FA, supplemented as canola oil (CO), on body weight (BW), daily gain, and immunological parameters of preweaning Holstein calves. The study was conducted from September to December 2019. Calves were randomly allocated to a control group (n = 15; BW = 36.2 ± 1.5 kg; mean ± SEM) supplemented daily with 30 mL of CO and to an experimental group (n = 15; BW = 36.3 ± 1.5 kg) supplemented with 60 g of a product containing 30 g of FO. Both treatments were added to the MR during the morning feeding. All calves were fed 4 L of MR at 12.5% solids at 0700 and 1600 h for wk 1, 6 L from wk 2 to 7, and 3 L once daily (0700 h) during wk 8 until weaning (56 d). Blood samples were collected at 7, 14, 21, 28, 35, 42, 49, and 56 d of age for serum haptoglobin, TNF-α, IL-1ß, and protectin. Dry matter intake was recorded in all experimental calves daily. Seroneutralization titers to vaccination against viral diseases (infectious bovine rhinotracheitis, parainfluenza 3, bovine viral diarrhea, and bovine respiratory syncytial virus) were determined. Mixed models for repeated measures were developed to analyze variables over time. Seroneutralization titers were analyzed by the Kruskal-Wallis test. The other variables were compared by a generalized linear model. Serum FA profile at 35 d of age showed that FO supported higher concentrations of n-3 FA than CO. Final BW [65.2 vs. 62.0 kg, standard error of the mean (SEM) = 2.1 kg] and average daily gain (0.52 vs. 0.46 kg/d, SEM = 0.1 kg/d) tended to be higher for the FO than the CO group. An interaction of treatment × day for dry matter intake was observed, especially during weaning (2.17 kg vs. 1.94 kg, SEM = 0.158 kg, for FO and CO group, respectively). Blood lactate (mmol/L) was higher in the CO than in the FO group at d 7. Haptoglobin and IL-1ß were higher for the CO group on d 14 than the FO group. The TNF- α concentrations for the FO group were reduced over time, whereas the concentrations in the CO group remained constant. Protectin was higher in the FO group on d 14, but was lower on d 28, 35, and 49. Seroneutralization antibody titers postvaccination for the PI3 virus were higher for the FO than the CO group. In conclusion, calves supplemented with FO had lower concentrations of blood lactate, haptoglobin, IL-1ß and TNF-α than calves supplemented with CO during the study period. The FO supplementation had a higher DMI than CO supplementation. Results of this trial should be interpreted with caution due to the lack of a negative control group as well as the lower birth weight and growth rate observed under heat stress conditions.

The association of prepartum urine pH, plasma total calcium concentration at calving and postpartum diseases in Holstein dairy cattle.

The use of anionic salts to prevent milk fever in dairy cattle has been an effective nutritional strategy; however, the degree of acidification that determines the most acceptable productive responses and well-being of the cow is still a controversial topic. The objective of this study was to assess urine pH in prepartum Holstein cows fed anionic diets and determine its association with plasma total Ca, Mg, P, ß-hydroxyl-butyrate (BHB) concentrations at parturition and the occurrence of peripartum disorders. This investigation consisted of 2 studies. Study 1 was conducted on a grazing dairy. Between February and May 2019, 60 prepartum multiparous cows were tested for urine pH and plasma metabolite concentration at parturition. Total Ca, P, Mg and BHB at day 1 in milk (DIM) were assessed and statistically analyzed by ANOVA (models for polynomial regression). Study 2 was conducted on a drylot dairy farm. Between July 2018 and January 2019, 203 cows were evaluated for urine pH and followed-up for 30 DIM to obtain the incidence of dystocia, stillbirths, milk fever, retained fetal membranes, metritis, clinical mastitis and ketosis. Cows were categorized based on their last urine pH as group 1: pH > 7.0 (n = 135); group 2: pH between 6.0 and 7.0 (n = 46) and group 3: pH < 6.0 (n = 22). A logistic regression model for each health event was conducted considering urine pH group as the main effect. Urine sample was collected at 2.71 ±â€¯2.84 days before parturition. In study 1, there was a quadratic effect of urine pH on total Ca. Total Ca concentration was higher between urine pH 6.0 and 7.0, while decreasing below pH 6.0 and above pH 7.0. There was a trend (P = 0.11) for a quadratic effect of urine pH on the concentration of plasma BHB at parturition. ß-Hydroxyl-butyrate was lower approximately between urine pH 6.5 and 7.5. In study 2, the odds for a stillborn in cows with urine pH < 6.0 was 2.39 (95% CI = 1.06-5.40) times the odds for a stillborn in cows with urine pH ≥ 7.0. There was no association between urine pH and the other diseases. In conclusion, cows with prepartum urine pH < 6.0 and >7.0 had lower concentration of plasma total Ca and tended to have a higher concentration of BHB. Cows with urine pH < 6.0 had a higher incidence of stillbirths than cows with urine pH > 7.0.

Lying and stepping behaviors around corrective or therapeutic claw trimming.

The objectives of this study were to test differences in lying and stepping behaviors in lactating cows with altered gait or other signs of pain in the day preceding either corrective claw trimming (CCT) or therapeutic claw trimming (TCT; d -1), including healthy controls (HC) as a reference group, and to analyze the associations between categories of lying and stepping behaviors at d -1 and subsequent submission to CCT or TCT (d 0). In addition, we characterized variations in lying and stepping behaviors within 7 d relative to CCT or treatment for a specific foot condition (TCT). A total of 310 Holstein cows in a USDA-certified organic dairy herd in northern Colorado were enrolled. Cows were affixed with an accelerometer (IceQube, IceRobotics) below the fetlock of one hind leg at 12 ± 8 d in milk and monitored for 6 mo for daily activity [lying time (LY; min/d), lying bouts (LB; no./d), and steps (ST; no./d)]. Cows with altered gait or other signs of pain were submitted to claw trimming (CT; d 0) and differentiated as receiving only corrective interventions (CCT) or as being treated for a lameness disorder (TCT). Concurrent activity from cows not submitted to CT was considered a reference from HC. Daily averages for each activity within ±7 d relative to CT were compared among the 3 groups using linear mixed models (PROC MIXED of SAS; SAS Institute Inc.). Logistic regression analyses with a logit link function (PROC GLIMMIX of SAS) were used to test the associations between categories (lower quartile or less, interquartile range, and higher quartile or greater) of lying and stepping behaviors (LY, LB, and ST) preceding CT (d -1) and subsequent submission to CCT or TCT (d 0). On d -1, average (± standard error) LY was greater in TCT (631 ± 34 min/d) than in CCT (568 ± 22 min/d) and HC (581 ± 13 min/d). Overall, LB were not statistically different among groups (CCT = 18.7 ± 0.02 bouts/d; TCT = 19.6 ± 1.17 bouts/d; and HC = 19.1 ± 0.43 bouts/d). Daily ST were lower in TCT (1,810 ± 126 steps/d) than in CCT (2,803 ± 63 steps/d) and HC (2,542 ± 103 steps/d). The odds (confidence interval in parentheses) of TCT were greater in cows in the high LY [3.27 (2.47-4.33)] and high LB [2.31 (1.77-3.02)] categories at d -1. The low ST category was associated with increased odds of CCT [3.32 (1.96-5.65)] and TCT [8.48 (6.49-11.1)]. We concluded that, under the current experimental conditions, cows requiring TCT evidenced greater overall LY and lower daily ST the day before CT. However, these differences were not consistent throughout days in milk categories within the lactation. Type of CT was partially associated with category of activity the day preceding CT, and the associations were more evident for TCT. The magnitude of the behavioral changes before and after CT and the recovery times posttreatment to approach HC behavioral levels varied depending on specific conditions.

Efficacy of estradiol or GnRH in combination with progesterone intravaginal devices to control the follicular wave dynamics and resulting fertility in lactating dairy cows.

This study was conducted to compare efficacy of treatments with EB or GnRH and different quantities of exogenous progesterone (P4) for synchronization of time of ovulation on follicular growth and pregnancy in lactating dairy cows. In Experiment 1, 40 cows were treated with EB or GnRH and 1.9 or 3.0 g of P4 via progesterone-containing intravaginal devices (IVPD; D0), two doses of PGF2α on D7, GnRH on D9, and TAI on D10. In Experiment 2, 1,440 cows were treated with EB or GnRH and 1 g IVPD on D0, cloprostenol, eCG and EB on D7. Cows in estrus by 48 h were AIDE, and non-estrous cows were administered GnRH and TAI 60 h after IVPD removal. Non-estrous cows were AIDE 72 h after IVPD removal. In Experiment 1, P4 was greater on D7 for cows treated with GnRH than those treated with EB. The dominant follicle was larger for cows treated with GnRH than those treated with EB. In Experiment 2, for estrous cows, pregnancy per AI was greater in cows AI at 48 h compared to 60 h after IVPD removal for cows treated with GnRH, and greater with AI at 60 h after IVPD removal compared to 48 h in EB-treated cows. In non-estrous cows, there was no effect on pregnancy. In conclusion, treatment with GnRH compared with EB resulted in increased P4 regardless of amount of exogenous P4, and there were differential proportions of estrous cows pregnant depending on time of AI after IVPD removal.

Associations between postpartum diseases and milk yield and changes in body condition between drying off and parturition of dairy cows in Argentina.

Aims: To assess the change in body condition score (BCS) during the early and late dry periods and its association with postpartum diseases and milk yield in grazing dairy cows from central Argentina. Methods: BCS assessments during the dry period, and cow health and milk production records up to 90 days in milk (DIM), were collated for cows from 28 farms at monthly visits between 2007 and 2008. Cows were categorised into four groups; those in Group 1 (n=7,067) maintained or gained BCS during the early and late dry periods; Group 2 (n=2,615) maintained or gained BCS during the early dry period and lost BCS during the late dry period; Group 3 (n=1,989) lost BCS during the early dry period and maintained or gained BCS during the late dry period; and Group 4 (n=5,144) lost BCS during the early and late dry periods. Results: Cows in Group 1 had reduced odds of having retained fetal membranes (RFM), metritis, and clinical mastitis up to 90 DIM than cows in Group 2 (p<0.001), but the odds of disease were similar to cows in Group 3. The odds of having RFM or clinical mastitis tended to be lower in cows in Group 1 than cows in Group 4 (p=0.08). The odds of cows being culled or dying during the first 90 DIM were lower for cows in Group 1 than for those in Groups 2, 3, and 4 (p≤0.05). Mean accumulated milk yield up to 90 DIM was higher in cows in Group 1 than Group 2 and Group 4 (p<0.001), but was similar to that of cows in Group 3 (p=0.28). Conclusions and clinical relevance: Cows that lost BCS during the late dry period had increased odds of being diagnosed with several postpartum diseases and had decreased milk yield compared to cows that maintained or gained BCS during the entire dry period. Loss of BCS during any stage of the dry period was also associated with increased incidence of culling or death during the first 90 DIM. These results should raise awareness among dairy cattle producers of the importance of properly managing cow body condition during the dry period, especially during the late dry period.

Safety and efficacy of a mesenchymal stem cell intramammary therapy in dairy cows with experimentally induced Staphylococcus aureus clinical mastitis.

Although, antibiotics are effective in the treatment of bovine mastitis, they do not address the regeneration of mammary glandular tissue and have been associated to the increment in antimicrobial resistance worldwide. Considering the necessity of alternative therapies for this disease of high economic impact and the reported regenerative and antibacterial effects of mesenchymal stem cell (MSCs), we evaluated the safety and efficacy of an allogenic MSC-based intramammary therapy in dairy cows with experimentally induced Staphylococcus aureus clinical mastitis. In a safety trial, heifers were inoculated intramammarily with a 2.5 × 107-suspension of bovine fetal AT-MSCs on experimental days 1 and 10. Animals were evaluated clinically on a daily basis during a 20-day experimental period and blood samples were collected for hemogram determination and peripheral blood leukocytes (PBLs) isolation. In an efficacy trial, Holstein Friesian cows were inoculated with S. aureus and treated intramammarily with vehicle (NEG; days 4 and 10), antibiotics (ATB; days 4 and 5) or a suspension of 2.5 × 107 AT-MSCs (MSC; days 4 and 5). Cows were clinically evaluated daily and milk samples were collected for somatic cell count (SCC) and colony forming units (CFU). Blood samples were collected for serum haptoglobin and amyloid A determination. Intramammary administration of two doses of bovine fetal AT-MSCs in healthy cows did not induce changes in clinical or hematological variables, and gene expression profiles in PBLs associated to activation (CD4, CD8, CD25, CD62L and CD69) and proinflammatory cytokines (CCL2, CCL5, IL2, CXCL3, IFNγ, and TNFα). Quarters of MSC group of cows had similar SCC log/mL in milk compared to infected quarters of ATB or NEG cows. However, quarters of MSC cows had lower CFU log/mL in milk compared to quarters of NEG cows. Intramammarily inoculation of repeated doses of 2.5 × 107 allogenic AT-MSCs did not induce clinical or immunological response in healthy cows. Moreover, MSC-intramammary treatment reduced bacterial count in milk of cows with S. aureus clinical mastitis compared to untreated cows. This work provides initial evidence for the safety and efficacy of an allogenic MSC-based intramammary therapy for the treatment of bovine mastitis.

Efficacy of non-antibiotic treatment options for digital dermatitis on an organic dairy farm.

The objective of this study was to evaluate the efficacy of two non-antibiotic treatment options for digital dermatitis (DD) on an organic certified dairy farm. A randomized clinical trial was conducted using 70 multiparous Holstein cows with an early DD lesion at a USDA certified organic dairy farm in Northern Colorado, USA. Cows were enrolled in the study based on the presence of early DD lesions (scores M1 and M2) and randomly assigned to one of three treatments: (1) topical application of copper sulfate and iodine (CUI); (2) topical application of honey and iodine (HOI); and (3) control subject to no treatment (CON). Cows were evaluated at enrolment and on days 3, 12, 28, and 120 post treatment for pain and lesion size and received a locomotion and a lesion score. Cure was defined as the transition from active to non-active stages (M1/M2 to M0 or M4). The formulations had variable effects on the treatment of DD. The cure rate was numerically higher for CUI on all follow up days. The proportion of cows experiencing pain on d3 after treatment was greater in CON, followed by HOI and CUI. However, this proportion increased in HOI during the follow up period. The CUI group had a greater reduction in lesion size and larger lesions persisted in HOI. Non-antibiotic treatment formulations were partially effective in the treatment of DD in organic dairy cows: The two non-antibiotic formulations resulted in an earlier transition to mature lesions compared with the control group. The CUI combination was the most effective treatment in reducing lesion size, pain, and lameness in affected cows. However, this combination had short-term efficacy, which did not persist throughout the duration of the study. The HOI combination produced only transient reduction in lesion size.

Use of milk electrical conductivity for the differentiation of mastitis causing pathogens in Holstein cows.

Mastitis is one of the most prevalent and costly diseases in dairy cattle. Key components for adequate mastitis control are the detection of early stages of infection, as well as the selection of appropriate management interventions and therapies based on the causal pathogens associated with the infection. The objective was to characterize the pattern of electrical conductivity (EC) in milk during intramammary infection, considering specific mastitis-causing pathogen groups involvement. Cows (n = 200) identified by an in-line mastitis detection system with a positive deviation ≥15% in the manufacturer's proprietary algorithm for EC (high electrical conductivity (HEC)) were considered cases and enrolled in the study at the subsequent milking. One control (CON) cow, within normal ranges for EC, was matched to each case. A composite milk sample was collected aseptically from each cow for bacteriological culture. Milk yield (MY) and EC were recorded for each milking during ±7 days relative to enrollment. Milk cultures were categorized into gram positive (GP), gram negative (GN), other (OTH) and no growth (NOG). Data were submitted for repeated-measures analysis with EC as the dependent variable and EC status at day -1, bacteriological culture category, parity number, stage of lactation and days relative to sampling as main independent variables. Average (± standard error (SE)) EC was greater in HEC than in CON cows (12.5 ± 0.5 v. 10.8 ± 0.5 mS/cm) on the day of identification (day -1). Milk yield on day -1 was greater in CON than in HEC (37.6 ± 5.1 v. 33.5 ± 5.2 kg). For practical management purposes, average EC on day -1 was similar for the different bacteriological culture categories: 11.4 ± 0.6, 11.7 ± 0.5, 12.3 ± 0.8 and 11.7 ± 0.5 mS/cm in GN, GP, OTH and NOG, respectively. Parity number was only associated with day -1 EC in HEC group, with the greatest EC values in parity 3 (12.3 ± 0.3 mS/cm), followed by parity 2 (11.9 ± 0.2 mS/cm), parity >3 (11.6 ± 0.5 mS/cm) and primiparous cows (11.2 ± 0.2 mS/cm). An effect on EC for the interaction of day relative to identification by pathogen gram category was observed. The same interaction effect was observed on daily MY. Overall, the level of variation for MY and EC between- and within-cows was substantial, and as indicated by the model diagnostic procedures, the magnitude of the variance in the cows in the CON group resulted in deviations from normality in the residuals. We concluded that characteristic temporal patterns in EC and MY in particular pathogen groups may provide indications for differentiation of groups of mastitis-causing pathogens. Further research to build detection models including EC, MY and cow-level factors is required for accurate differentiation.

Genome-wide study to detect single nucleotide polymorphisms associated with visceral and subcutaneous fat deposition in Holstein dairy cows.

Excessive abdominal fat might be associated with more severe metabolic disorders in Holstein cows. Our hypothesis was that there are genetic differences between cows with low and high abdominal fat deposition and a normal cover of subcutaneous adipose tissue. The objective of this study was to assess the genetic basis for variation in visceral adiposity in US Holstein cows. The study included adult Holstein cows sampled from a slaughterhouse (Green Bay, WI, USA) during September 2016. Only animals with a body condition score between 2.75 and 3.25 were considered. The extent of omental fat at the level of the insertion of the lesser omentum over the pylorus area was assessed. A group of 100 Holstein cows with an omental fold <5 mm in thickness and minimum fat deposition throughout the entire omentum, and the second group of 100 cows with an omental fold ⩾20 mm in thickness and with a marked fat deposition observed throughout the entire omentum were sampled. A small piece of muscle from the neck was collected from each cow into a sterile container for DNA extraction. Samples were submitted to a commercial laboratory for interrogation of genome-wide genomic variation using the Illumina BovineHD Beadchip. Genome-Wide association analysis was performed to test potential associations between fat deposition and genomic variation. A univariate mixed linear model analysis was performed using genome-wide efficient mixed model association to identify single nucleotide polymorphisms (SNPs) significantly associated with variation in a visceral fat deposition. The chip heritability was 0.686 and the estimated additive genetic and residual variance components were 0.427 and 0.074, respectively. In total, 11 SNPs defining four quantitative trait locus (QTL) regions were found to be significantly associated with visceral fat deposition (P<0.00001). Among them, two of the QTL were detected with four and five significantly associated SNPs, respectively; whereas, the QTLs detected on BTA12 and BTA19 were each detected with only one significantly associated SNP. No enriched gene ontology terms were found within the gene networks harboring these genes when supplied to DAVID using either the Bos taurus or human gene ontology databases. We conclude that excessive omental fat in Holstein cows with similar body condition scores is not caused by a single Mendelian locus and that the trait appears to be at least moderately heritable; consequently, selection to reduce excessive omental fat is potentially possible, but would require the generation of predicted transmitting abilities from larger and random samples of Holstein cattle.

Effect of diets containing sulfate or chloride-based anionic salts, fed to grazing prepartum dairy cows, on concentrations of Ca in plasma, disease incidence and milk yield.

AIMS: To compare the effect of two partial mixed ration diets containing either sulfate or chloride-based anionic salts, fed to grazing prepartum dairy cows, on concentrations of Ca, Mg, P and beta-hydroxybutyrate (BHB) in plasma, disease incidence and milk yield during the early postpartum period. METHODS: The study was conducted on a seasonal-calving grazing dairy farm located in Benton (MO, USA). In August 2016, two groups of 200 cows were randomly assigned at 30 days before expected parturition to receive either a sulfate or chloride-based anionic diet, to achieve a dietary cation-anion difference (DCAD) of -25 mEq/kg DM. Cows were fed partial mixed rations to provide 50% of their DM intake, with 50% provided by grazing fescue pasture. Urine samples were collected from 20 cows from each group once a week for 5 weeks until calving for pH assessment. At parturition, 42 cows per group were selected at random and blood samples collected on Days 1, 2, 3 and 7 postpartum. The study continued until 100 cows per group calved. Health events and cumulative milk yield until 30 days postpartum were recorded. Concentrations of Ca, P, and Mg in plasma at Days 1, 2, 3, and 7, and BHB in plasma at Day 7 postpartum were assessed. RESULTS: Mean urine pH was lower in cows in the chloride than sulfate group (p≤0.06) on four of five sampling occasions. Mean concentrations of Ca in plasma on Day 1 were 2.00 (95% CI=1.94-2.05) and 1.91 (95% CI=1.88-1.97) mmol/L for the chloride and sulfate groups, respectively (p=0.15), and on Day 3 were 2.03 (95% CI=1.97-2.08) and 1.90 (95% CI=1.84-1.96) mmol/L, respectively (p=0.038). Concentrations of Mg, P and BHB in plasma and incidence of diseases postpartum were similar in both groups (p>0.05). There were no cases of clinical hypocalcaemia in either group. CONCLUSIONS AND CLINICAL RELEVANCE: Prepartum cows fed a partial mixed ration with a negative DCAD containing chloride-based anionic salts and free access to a fescue pasture had lower prepartum urine pH, and higher concentrations of total Ca in plasma during the first week postpartum, than cows receiving a partial mixed ration containing sulfate-based anionic salts.

Technical note: Evaluation of fine needle aspiration cytology for the diagnosis of fatty liver in dairy cattle.

Fatty liver is a common condition affecting dairy cattle during the periparturient period, characterized by a pathological accumulation of triglycerides (TG) in the hepatocytes. The objective of this study was to evaluate the diagnostic potential of fine needle aspiration cytology in fresh liver specimens using liver TG concentrations as a gold standard. Fifty-seven liver samples from Holstein cows were collected during processing at a slaughterhouse. Tissue and fine needle aspirate samples were obtained from the parietal upper portion of the caudate lobe. Two samples of liver tissue were collected with a 16 gauge × 15 cm biopsy needle for histological and TG concentration assessment. A third sample was collected for cytology using an 18 gauge × 5.08 cm needle. The contents of the needle were transferred to a glass slide, spread, and air-dried. Liver samples were assayed by colorimetry/fluorimetry to determine TG concentrations. Concentrations of TG <2% were considered normal. Histological and cytological evaluations were conducted by 2 different pathologists blind to the visual classification. Sensitivity (Se) and specificity (Sp) were calculated. Cytology had a Se and Sp of 73 and 85%, respectively. Histopathology had a Se and Sp of 45.9 and 100%, respectively. The likelihood of having higher scores for histopathology and cytology increased as a function of liver TG content (mg/g).

Enfermedad neumocócica invasiva por serotipo 38, reporte de caso clínico / Invasive pneumococcal disease by serotype 38, a clinical case report

La enfermedad neumocócica invasiva (ENI) es causa de morbimortalidad prevenible en pediatría. Con la introducción de vacunas antineumocócicas conjugadas disminuyó la prevalencia de ENI en 61,9% en los menores de 2 años, y se produjo un cambio en la distribución de serotipos y un aumento de ENI por serotipos no vaccinales. En este contexto, es relevante la vigilancia epidemiológica de los serotipos emergentes causantes de ENI en la población. Se presentará el caso de una lactante de 11 meses con diagnóstico de meningitis causada por neumococo serotipo 38, su evolución y consecuencias clínicas, y se realiza un análisis de la situación epidemiológica actual.

In pediatrics,invasive pneumococcal disease is a preventable cause ofmorbidity andmortality.The introduction of conjugated pneumococcal vaccines has reduced the prevalence of invasive pneumococcal disease by 61.9% in the under two's and has brought about a change in the distribution of serotypes and a rise in invasive pneumococcal disease caused by non-vaccine serotypes.This being the case,itis very importanttomonitorthe epidemiology ofthe emerging serotypes causing the disease in the population.We presentthe case of an 11 month old infant diagnosed with meningitis caused by serotype 38, describing his clinical course andclinical consequences; andweperforman analysis ofthepresent epidemiologica lsituation

Retrospective evaluation of milk production and culling risk following either surgical, toggle-pin suture or conservative treatment of left displaced abomasum in Chilean dairy cows.

AIMS To describe milk yield and culling risk in cows diagnosed with left displacement of abomasum (LDA) treated either conservatively, by right flank pyloric omentopexy, or rolling and toggling, compared with normal herdmates from four Chilean dairy herds. METHODS Historical records were obtained from four commercial dairy farms located in Central Chile for cows with a history of LDA between 2010 and 2012, and healthy herdmates. Cows with LDA were categorised into three groups: cows treated with right omentopexy (ST, n=58), cows treated by toggle suturing (TT, n=15) and cows treated conservatively (CT, n=56). Control cows (n=129) were selected from unaffected cows, matched by days in milk (DIM), parity and herd with affected cows. Groups were compared for risk of culling up to 300 DIM and for milk production up to 5 months of lactation using survival and Cox proportional hazard models and mixed models for repeated measures, respectively. RESULTS Compared with cows in the Control group, the risk of being culled up to 300 DIM was 9.1 (SE 0.62) times greater in ST cows, 10.4 (SE 0.68) times greater in TT cows, and 37.3 (SE 0.61) times greater in CT cows (p<0.01). In the first 5 months of lactation, compared with cows in the Control group, mean daily milk production was 23.3 (SE 1.5) kg less in ST cows, 15.3 (SE 1.6) kg less in TT cows, and 30.1 (SE 1.3) kg less in CT cows (p<0.001). CONCLUSIONS AND CLINICAL RELEVANCE Cows in four dairy herds in central Chile diagnosed and treated for LDA produced significantly less milk and had a higher risk of culling than healthy herdmates. Although cows treated surgically or with toggle suture never recovered to the extent of healthy cows, they produced more milk than cows treated conservatively. However, the retrospective nature of the data, the inclusion of only four herds and the non-random allocation to treatments means that these conclusions cannot be extrapolated to the overall dairy cattle population in Chile.

Hot topic: 16S rRNA gene sequencing reveals the microbiome of the virgin and pregnant bovine uterus.

We tested the hypothesis that the uterus of virgin heifers and pregnant cows possessed a resident microbiome by 16S rRNA gene sequencing of the virgin and pregnant bovine uterus. The endometrium of 10 virgin heifers in estrus and the amniotic fluid, placentome, intercotyledonary placenta, cervical lumen, and external cervix surface (control) of 5 pregnant cows were sampled using aseptic techniques. The DNA was extracted, the V4 hypervariable region of the 16S rRNA gene was amplified, and amplicons were sequenced using Illumina MiSeq technology (Illumina Inc., San Diego, CA). Operational taxonomic units (OTU) were generated from the sequences using Qiime v1.8 software, and taxonomy was assigned using the Greengenes database. The effect of tissue on the microbial composition within the pregnant uterus was tested using univariate (mixed model) and multivariate (permutational multivariate ANOVA) procedures. Amplicons of 16S rRNA gene were generated in all samples, supporting the contention that the uterus of virgin heifers and pregnant cows contained a microbiome. On average, 53, 199, 380, 382, 525, and 13,589 reads annotated as 16, 35, 43, 63, 48, and 176 OTU in the placentome, virgin endometrium, amniotic fluid, cervical lumen, intercotyledonary placenta, and external surface of the cervix, respectively, were generated. The 3 most abundant phyla in the uterus of the virgin heifers and pregnant cows were Firmicutes, Bacteroidetes, and Proteobacteria, and they accounted for approximately 40, 35, and 10% of the sequences, respectively. Phyla abundance was similar between the tissues of the pregnant uterus. Principal component analysis, one-way PERMANOVA analysis of the Bray-Curtis similarity index, and mixed model analysis of the Shannon diversity index and Chao1 index demonstrated that the microbiome of the control tissue (external surface of the cervix) was significantly different from that of the amniotic fluid, intercotyledonary placenta, and placentome tissues. Interestingly, many bacterial species associated with postpartum uterine disease (i.e., Trueperella spp., Acinetobacter spp., Fusobacteria spp., Proteus spp., Prevotella spp., and Peptostreptococcus spp.) were also present in the uterus of virgin heifers and of pregnant cows. The presence of 16S rRNA gene sequence reads in the samples from the current study suggests that the uterine microbiome is established by the time a female reaches reproductive maturity, and that pregnancies are established and maintained in the presence of a uterine microbiome.

Association between disease occurrence and fertility of dairy cows in three geographic regions of Chile.

The objective was to analyze the association between disease occurrence during early lactation and reproductive performance and survival of dairy cows in high-producing herds, under different management practices in three geographic regions of Chile. Data included 30,757 lactation records of cows calving from January 2013 to June 2014 in three different locations: Central (C) area (n = 6198 cows in eight herds), Central-South (CS) area(n = 17,234 cows in 12 herds), and South (S) area (n = 7325 cows in six herds). Data were analyzed using logistic regression and ANOVA, considering cow as the experimental unit. Covariables offered to the models included parity number, season of calving, cow and herd relative milk yield, geographic location, and management system. Average milk yield (305 ME) per cow were 12,091, 11,783, and 6852 kg for C, CS, and S regions, respectively. Time from calving to first service and time to conception were consistently greater for cows with at least one disease event within 50 days in milk (DIM), for cows that were reported lame, or for cows that had mastitis or metritis. The odds (95% confidence interval) of pregnancy at 150 DIM (P150) and the odds of survival until 150 DIM (S150) for cows that had at least one disease event within 50 DIM were 0.84 (0.79-0.91) times the odds of pregnancy and 0.25 (0.22-0.28) times the odds of survival for healthy cows. The odds of P150 for cows located in the C and CS areas were 1.56 (1.36-1.80) and 1.16 (1.04-1.30) times the odds of P150 for cows in the S area. The odds of S150 for cows located in the C and CS areas were 0.48 (0.37-0.62) and 0.54 (0.42-0.67) times the odds of S150 for cows in the S area. These data suggested that cow health status and geographic location are significantly associated with reproductive performance and survival in this population of Chilean dairy cows.