Identification of genetic markers associated with hyperketonemia patterns in early lactation Holstein cows.

Ketosis, evidenced by hyperketonemia with elevated blood ß-hydroxybutyrate (BHB) levels, is a significant metabolic disorder of dairy cattle, typically diagnosed within the first 6 weeks post-calving when high energy levels are essential to milk production. Our study aimed to identify genetic markers linked to hyperketonemia (HYK) patterns in Holstein cows during early lactation and compare these to HYK-negative cows. We screened 964 cows for HYK using a threshold of BHB ≥1.2 mmol/L during the first 2 weeks postpartum (screening period, SP). Cows that tested negative initially were retested the following week. Cows were deemed HYK-negative (CON group) if BHB levels were below 1.2 mmol/L in both tests, while those with BHB levels exceeding this threshold at any test were treated and classified as HYK-positive (HYK+). Post-treatment, HYK+ cows were monitored for two-week follow-up period (FP) and classified based on their recovery: cured (CUR; consistently low BHB), recurrent (REC; fluctuating BHB levels), severe (SEV; high initial BHB that decreased), or chronic (CHR; persistently high BHB). Using 489 cows that were genotyped, a GWAS was conducted using GCTA software, revealing significant associations of several SNPs across different HYK patterns when compared to the CON group. These SNPs were primarily linked to genes affecting milk traits and were enriched in biological pathways relevant to protein glycosylation, inflammatory response, glucose homeostasis, and fatty acid synthesis. Our findings highlight genomic regions, potential candidate genes, and biological pathways related to ketosis, underscoring potential targets for improving health management in dairy cattle. These insights could lead to better strategies for managing ketosis through genetic selection, ultimately enhancing dairy cattle welfare and productivity. Further research with a larger number of cows is recommended to validate these findings and help confirm the implicated SNPs and genes.

Perspective: Transient postparturient hypocalcemia-A lactation-induced phenomenon of high-producing dairy cows.

Milk fever is one of the most historically relevant diseases of dairy cows. It is caused by tremendous calcium (Ca) expenditure at the initiation of lactation, so severe that cows can no longer stand and, if left untreated, die. Fortunately, through prepartum nutritional improvements, this version of clinical hypocalcemia is rare in the United States. Nonetheless, the opinion that all versions of postpartum hypocalcemia are detrimental remains pervasive, which is particularly significant given that 50% of cows are subclinically hypocalcemic after calving. This has led to a variety of available management and treatment strategies, ranging from prepartum dietary programs to postpartum Ca gels and boluses, targeted at preventing hypocalcemia in dairy cows. Recent research has determined that postpartum dairy cows can experience different types of subclinical hypocalcemia: transient, persistent, or delayed. We now know cows experiencing transient hypocalcemia as part of the homeorhetic adaptation to lactation are the highest milk producers in modern dairy herds, whereas cows with hypocalcemia several days after calving experience disease and losses in milk production. Therefore, it is wrong to assume all postpartum hypocalcemia is detrimental and that treatment of all cases is considered necessary and beneficial. Research indicates that milk synthesis at the onset of lactation contributes to immediate postpartum hypocalcemia, and that the mammary gland is a critical factor in management of Ca homeostasis. However, cows differ in their ability to manage this phenomenon, and it is possible that immediate postpartum influences such as dry matter intake, inflammation, and immune activation affect appropriate Ca regulation in the days following calving.

Hyperketonemia: A Marker of Disease, a Sign of a High-Producing Dairy Cow, or Both?

This review covers the history and nomenclature of ketosis, the source and use of ketones in transition cows, and the controversial role of hyperketonemia's association with health and production outcomes in dairy cows. With the goal of assisting veterinarians with on-farm diagnostic and treatment methods, the authors present current and evolving means of direct and indirect hyperketonemia detection as well as a summary of treatment modalities and their efficacy. They encourage veterinarians to include hyperketonemia testing as part of their routine physical examinations and contemplate day in milk at hyperketonemia diagnosis when designing treatment and management strategies.

Considerations in the Diagnosis and Treatment of Early Lactation Calcium Disturbances.

This article reviews the history of clinical hypocalcemia and the evolving definition of subclinical hypocalcemia, targeting a concept for consideration that not all hypocalcemia is negative. With a goal of presenting bovine practitioners information to assist with individual animal hypocalcemia diagnosis and treatment as well as herd-level monitoring and prevention, we present current methods of direct calcium measurement, therapeutic interventions for clinical hypocalcemia, and postpartum calcium supplementation options and their efficacy. We encourage veterinarians to understand calcium dynamics in the immediate postpartum period and evaluate how individual cow therapy and herd prevention protocols can assist with supporting calcium regulation.

The association of hyperketonemia with fecal and rumen microbiota at time of diagnosis in a case-control cohort of early lactation cows.

BACKGROUND: Many dairy cows experience a state of energy deficit as they transition from late gestation to early lactation. The aims of this study were to 1) determine if the development of hyperketonemia in early lactation dairy cows is indicated by their gut microbiome, and 2) to identify microbial features which may inform health status. We conducted a prospective nested case-control study in which cows were enrolled 14 to 7 days before calving and followed through their first 14 days in milk (DIM). Hyperketonemic cows (HYK, n = 10) were classified based on a blood ß-hydroxybutyrate (BHB) concentration 1.2 mmol/L within their first 14 DIM. For each HYK cow, two non-HYK (CON, n = 20) cows were matched by parity and 3 DIM, with BHB < 1.2 mmol/L. Daily blood BHB measures were used to confirm CON cows maintained their healthy status; some CON cows displayed BHB 1.2 mmol/L after matching and these cows were reclassified as control-HYK (C-HYK, n = 9). Rumen and fecal samples were collected on the day of diagnosis or matching and subjected to 16S rRNA profiling. RESULTS: No differences in taxa abundance, or alpha and beta diversity, were observed among CON, C-HYK, and HYK health groups for fecal microbiomes. Similar microbiome composition based on beta diversity analysis was detected for all health statuses, however the rumen microbiome of CON and HYK cows were found to be significantly different. Interestingly, highly similar microbiome composition was observed among C-HYK cow rumen and fecal microbiomes, suggesting that these individual animals which initially appear healthy with late onset of hyperketonemia were highly similar to each other. These C-HYK cows had significantly lower abundance of Ruminococcus 2 in their rumen microbiome compared to CON and HYK groups. Multinomial regressions used to compute log-fold changes in microbial abundance relative to health status were not found to have predictive value, therefore were not useful to identify the role of certain microbial features in predicting health status. CONCLUSIONS: Lower relative abundance of Ruminococcus 2 in C-HYK cow rumens was observed, suggesting these cows may be less efficient at degrading cellulose although the mechanistic role of Ruminococcus spp. in rumen metabolism is not completely understood. Substantial differences in fecal or rumen microbiomes among cows experiencing different levels of energy deficit were not observed, suggesting that hyperketonemia may not be greatly influenced by gut microbial composition, and vice versa. Further studies using higher resolution -omics approaches like meta-transcriptomics or meta-proteomics are needed to decipher the exact mechanisms at play.

Ophthalmic disorders in a referral population of seven breeds of brachycephalic dogs: 970 cases (2008-2017).

OBJECTIVE: To evaluate the frequency of ophthalmic disorders in 7 brachycephalic dog breeds referred to an academic veterinary ophthalmology service. ANIMALS: 970 client-owned dogs of 7 brachycephalic breeds that were evaluated by the ophthalmology service in a veterinary teaching hospital from January 2008 through December 2017. PROCEDURES: Medical records of 7 brachycephalic breeds (ie, Boston Terriers, English Bulldogs, French Bulldogs, Lhasa Apsos, Pekingese, Pugs, and Shih Tzus) were reviewed to collect data regarding patient signalment, ophthalmic diagnoses, affected eyes, and number and dates of visits. RESULTS: Median age at the first examination was 7 years (range, 23 days to 22 years). The number of dogs seen for a first examination increased with age. Corneal ulcers, keratoconjunctivitis sicca, corneal pigmentation, immature cataracts, and uveitis were each diagnosed in ≥ 100 dogs and represented 40.4% (1,161/2,873) of all diagnoses. On the basis of anatomic location, 66.3% (1,905/2,873) of all disorders were located in either the cornea (1,014/2,873 [35.2%]) or adnexa (891/2,873 [31%]). There was a significant difference in breed proportion in the study population; of the 7 breeds studied, Shih Tzus (34.3% [333/970]), Pugs (20.8% [202/970]), and Boston Terriers (16.6% [161/970]) were the most prevalent breeds. The frequency of some diseases within the referral population was associated with breed. CONCLUSIONS AND CLINICAL RELEVANCE: Findings suggested that the most prevalent disorders for the brachycephalic breeds in this ophthalmic referral population were corneal ulcers, keratoconjunctivitis sicca, corneal pigmentation, immature cataracts, and uveitis. Although all dogs shared brachycephalic features, the frequency of specific ophthalmic diseases varied between breeds.

Symposium review: Transition cow calcium homeostasis-Health effects of hypocalcemia and strategies for prevention.

The effects of subclinical hypocalcemia have been explored in numerous observational and mechanistic studies in recent years. Besides obvious, well-known effects on muscle contractility, the role of Ca with respect to immune function and intermediary metabolism explains the contribution of subclinical hypocalcemia to the development of several diseases observed in early lactation and underlines its importance in high-performing dairy cows. The present review aims at integrating recent scientific progress, such as discoveries about the role of the mammary gland in regulating bone mobilization, into generally accepted aspects of the endocrine control of Ca homeostasis. We will discuss Ca transport mechanisms through absorption, resorption, secretion, and mobilization, as well as the physiological regulation of Ca through parathyroid hormone, 1,25-dihydroxyvitamin D, fibroblast growth factor 23, and serotonin, in addition to dietary mineral requirements. To improve hypocalcemia prevention strategies, our knowledge of the physiological mechanisms necessary to maintain normocalcemia and their endogenous regulation should be combined with data derived from herd-level studies. Using such studies, we will discuss prepartum nutritional strategies aimed at reducing the incidence of subclinical hypocalcemia, as well as options for postpartum Ca supplementation and their effects on early-lactation health and production. Especially in respect to approaches that might interfere with endogenous adaptation processes, such as supplementation with vitamin D metabolites or large doses of Ca, a thorough understanding of the underlying mechanisms that might induce unwanted hypocalcemia rebound effects will be crucial to ameliorate our future management of transition cows. Continued efforts by researchers to understand the interaction of Ca homeostasis with prevention strategies is necessary to optimize cow health and support copious milk production.

Blood ß-hydroxybutyrate concentrations and early lactation management strategies on pasture-based dairy farms in Colombia.

The increasing global demand for food requires sustainable solutions to close the gap in agricultural yield between industrialized and non-industrialized countries. Our objectives in this cross-sectional study were to: 1) characterize farm populations, milk yield, and early lactation management strategies of dairy cows in three different regions of Colombia, and 2) determine the association of these management strategies with blood ß-hydroxybutyrate (BHB) concentrations in the first 42 days in milk (DIM). Dairy herds (n = 56) in the Antioquia, Caldas, and Cundinamarca regions of Colombia were visited once from May through July 2018. A survey was administered to farm owners to collect demographic, management, and herd nutrition information. Blood samples from dairy cows (n = 880) between calving and 42 DIM were used to measure blood BHB concentration. Associations between management and nutritional strategies and blood BHB concentration were examined using mixed models. Prevalence of hyperketonemia was calculated as the number of samples with BHB concentration ≥1.2 mmol/L divided by the total number of samples. The estimated diet composition for early lactation dairy cows was 65.5% pasture and 31.8% commercial concentrates. The farm median milk yield, protein concentration, and fat concentration were 21.0 kg (range = 13.1-36 kg), 3.2% (range = 2.7-4.1%), and 3.5% (range = 3.0-4.1%), respectively. Milk yield least squares means (95% confidence interval; CI) differed by region: 21.7 (20.3, 23.2), 18.5 (17.0, 20.2), and 20.3 (18.5, 22.4) kg in Antioquia, Caldas, and Cundinamarca, respectively. Median blood BHB concentration was 0.5 and ranged from 0.1-4.4 mmol/L; blood BHB concentration was not different among the three regions. Pasture fertilization, increased parity, and BCS were associated with changes in blood BHB concentration. The overall prevalence of hyperketonemia was 4.5%. Geographical region affected the prevalence of hyperketonemia at 2.5%, 4.0%, and 10.2% in Antioquia, Caldas, and Cundinamarca, respectively. Mean stocking density (95% CI) was greater in Cundinamarca than Antioquia or Caldas at 3.3 (2.2, 5.0), 2.8 (2.1, 3.9) and 1.7 (1.2, 2.6) animals per ha, respectively, and was associated with hyperketonemia prevalence. Farms that abruptly stop milking cows at dry-off had 80% of the hyperketonemia events in the study. Pasture-based dairies in Colombia had lower blood BHB concentrations and estimated milk yield compared with confined production systems in temperate zones. However, geographical region, stocking density, and abrupt cessation of milking at dry-off were associated with prevalence of hyperketonemia in pasture-based dairies.

Longitudinal Phenotypes Improve Genotype Association for Hyperketonemia in Dairy Cattle.

The objective of our study was to identify genomic regions associated with varying concentrations of non-esterified fatty acid (NEFA), ß-hydroxybutyrate (BHB), and the development of hyperketonemia (HYK) in longitudinally sampled Holstein dairy cows. Our study population consisted of 147 multiparous cows intensively characterized by serial NEFA and BHB concentrations. To identify individuals with contrasting combinations in longitudinal BHB and NEFA concentrations, phenotypes were established using incremental area under the curve (AUC) and categorized as follows: Group (1) high NEFA and high BHB, group (2) low NEFA and high BHB), group (3) low NEFA and low BHB, and group (4) high NEFA and low BHB. Cows were genotyped on the Illumina Bovine High-density (777K) beadchip. Genome-wide association studies using mixed linear models with the least-related animals were performed to establish a genetic association with HYK, BHB-AUC, NEFA-AUC, and the comparisons of the 4 AUC phenotypic groups using Golden Helix software. Nine single-nucleotide polymorphisms were associated with high longitudinal concentrations of BHB and further investigated. Five candidate genes related to energy metabolism and homeostasis were identified. These results provide biological insight and help identify susceptible animals thus improving genetic selection criteria thereby decreasing the incidence of HYK.

Udder and teat conformational risk factors for elevated somatic cell count and clinical mastitis in New York Holsteins.

Our primary objective was to identify udder and teat conformational risk factors associated with the occurrence of elevated somatic cell count (SCC) and clinical mastitis using a prospective cohort study design with careful assessment of exposure and disease outcomes. Mastitis prevalence was evaluated by parity across 6 sampling periods representing key physiological transitions during lactation: 0-1 day in milk (DIM), 3-5 DIM, 10-14 DIM, 50-60 DIM, 90-110 DIM, and 210-230 DIM. Cows were scored for front and rear teat length, width, end shape, and placement, fore udder attachment, udder cleft, udder depth, rear udder height, and rear udder width. Two independent multivariable logistic regression models were used to generate odds ratios (OR) for elevated SCC (≥ 200,000 cells/ml) and farm-diagnosed clinical mastitis. We identified that loose fore udder attachment (reference level: strong fore udder attachment, OR = 2.1, 95% confidence interval (CI) = 1.2-3.8) and flat teat end shape (reference level: round teat end shape, OR = 1.4, 95% CI = 1.1-1.9) increased the odds of an elevated SCC event, whereas a negative California Mastitis Test score at 0-1 DIM decreased the odds of an elevated SCC event (OR = 0.6, 95% CI = 0.4 to 0.8). Loose fore udder attachment (reference level: strong fore udder attachment, OR = 3.7, 95% CI = 1.3-10.7), flat teat end shape (reference level: round teat end shape, OR = 1.5, 95% CI = 1.0-2.4), low rear udder height (reference level: intermediate rear udder height, OR = 2.8, 95% CI = 0.3-6.2), and increasing rear teat width (OR = 2.2, 95% CI = 1.2-4.4) heightened the odds of developing clinical mastitis. We identified that within our study cohort, loose fore udder attachment and flat teat ends had an important association with increased odds of both an elevated SCC event and clinical mastitis diagnosis. The identification of these udder and teat conformational risk factors for mastitis can provide farmers an effective and inexpensive tool to manage mastitis.

A longitudinal study of digital cushion thickness and its function as a predictor for compromised locomotion and hoof lesions in Holstein cows.

Lameness is a major animal welfare and economic issue for the dairy industry and is a challenge to overcome due to multifaceted causes. Digital cushion thickness (DCT) is a strong predictor of lameness and is phenotypically associated with incidence of claw horn disruption lesions (CHDL; sole ulcers and white line disease). We hypothesized that DCT varies between digits and across lactation within the cow. This variation could be characterized to predict the occurrence of CHDL or compromised locomotion. BCS, visual locomotion score (VLS), DCT, and presence or absence of lesions were collected at 4 time points: <40 d prepartum (DPP), 1 to 30 d in milk (DIM), 90 to 120 DIM, and ≥255 DIM for 183 commercial Holstein cows enrolled in the study. Cows underwent digital sonographic examination for the measurement of DCT evaluated at the typical sole ulcer site beneath the flexor tuberosity for the right front medial and lateral digits and right hind medial and lateral digits. Factors such as parity number and stage in lactation were obtained from farm management software (DairyComp 305; Valley Agricultural Software, Tulare, CA). Cows were grouped by parity: primiparous (parity = 1) or multiparous (parity ≥ 2). The prevalence of CHDL among time points ranged from 0% to 4.2% for primiparous cows vs. 2.5% to 25% for multiparous cows, whereas the prevalence of lameness based on VLS of 3 to 5 ranged from 1.7% to 8.3% for primiparous cows vs. 12.7% to 33% for multiparous cows. DCT varied within primiparous and multiparous cows based on stage of lactation and digit (P < 0.05) and was thicker for both parity groups prior to dry off (≥255 DIM) and thinnest prior to calving (<40 DPP) and after peak lactation (90 to 120 DIM). The DCT of the front medial digit was thickest for primiparous heifers, whereas the hind lateral digit was thickest for multiparous cows. The DCT of the hind medial digit was thinnest for both parity groups. Parity group and DCT of the hind lateral digit <40 DPP were important predictors of CHDL (P < 0.05), whereas parity group and DCT of the hind lateral digit and front lateral digit at 1 to 30 DIM were key predictors of VLS lameness (P < 0.05). These results may help identify animals with higher odds of developing these diseases by highlighting key time points and specific digits of importance for monitoring. In addition, it improves our biological understanding of the relationship between DCT and lameness.

Herd-Level Monitoring and Prevention of Displaced Abomasum in Dairy Cattle.

Displaced abomasum (DA) is a postpartum disease that causes significant economic losses in the dairy industry. Abomasal atony and excessive production of gas have been reported as prerequisites for the development of DA. The exact cause of DA is unknown, yet infectious and metabolic disease, diet composition and physical form, cow comfort, and management of dairy cows during the transition period have been associated with the occurrence of this disorder. This review article discusses different factors that lead to the development of DA and strategies for monitoring DA and its comorbidities at the herd level.

Field trial of 2 calcium supplements on early lactation health and production in multiparous Holstein cows.

Our objectives were to measure serum Ca concentrations in the first 48 h postpartum in cows supplemented with oral Ca or subcutaneous Ca and nonsupplemented cows and evaluate the effect of these treatments on the incidence of metritis, displaced abomasum, mastitis, and early lactation disease (any of the diseases milk fever, retained placenta, metritis, or displaced abomasum), removal from the herd, pregnancy to first insemination, and average daily milk yield for the first 10 wk of lactation. We conducted 2 experiments on 1 commercial herd in New York State. In experiment 1, multiparous Holstein cows (n = 30) were blocked by parity (2 and ≥3) and sequentially assigned at calving to nontreated control (CON, n = 10), subcutaneous administration of 500 mL 23% Ca gluconate at calving (SC, n = 10), or administration of an oral Ca bolus containing 43 g of calcium at calving and again 12 h later (OB, n = 10). Blood was collected before treatment and at 1, 2, 4, 8, 12, 24, and 48 h thereafter for measurement of serum total Ca concentration. In experiment 2, 1,478 multiparous Holstein cows were sequentially assigned by calving date to the same 3 treatments (CON, n = 523; SC, n = 480; OB, n = 475). In experiment 1, SC cows had greater Ca concentrations from 1 through 12 h post-treatment and OB cows had greater Ca concentrations at 1 and 24 h post-treatment compared with CON cows. We found no difference in risk of metritis, displaced abomasum, early lactation disease diagnosis, or pregnancy to first insemination among treatments. Treatment with SC or OB had no effect on average daily milk yield compared with CON cows (CON = 46.7 kg; SC = 47.1 kg; OB = 47.0 kg). Cows treated with SC or OB that had a high relative herd milk rank in the previous lactation were almost half as likely to be diagnosed with mastitis in the first 60 DIM compared with CON cows [risk ratio (RR)SC = 0.57, RROB = 0.54]; however, we found no difference in risk of mastitis among treatments for cows with low relative herd milk rank. Second-parity cows fed a negative prepartum dietary cation-anion difference ration and treated with SC or OB were more likely to be removed from the herd than CON cows (RRSC = 3.91, RROB = 4.72); this difference was not observed in second-parity cows fed a neutral prepartum dietary cation-anion difference ration or in parity ≥3 cows. Although Ca supplementation increased serum Ca, this effect did not greatly improve milk production or health and reproductive outcomes.

Characterization of the resistome in manure, soil and wastewater from dairy and beef production systems.

It has been proposed that livestock production effluents such as wastewater, airborne dust and manure increase the density of antimicrobial resistant bacteria and genes in the environment. The public health risk posed by this proposed outcome has been difficult to quantify using traditional microbiological approaches. We utilized shotgun metagenomics to provide a first description of the resistome of North American dairy and beef production effluents, and identify factors that significantly impact this resistome. We identified 34 mechanisms of antimicrobial drug resistance within 34 soil, manure and wastewater samples from feedlot, ranch and dairy operations. The majority of resistance-associated sequences found in all samples belonged to tetracycline resistance mechanisms. We found that the ranch samples contained significantly fewer resistance mechanisms than dairy and feedlot samples, and that the resistome of dairy operations differed significantly from that of feedlots. The resistome in soil, manure and wastewater differed, suggesting that management of these effluents should be tailored appropriately. By providing a baseline of the cattle production waste resistome, this study represents a solid foundation for future efforts to characterize and quantify the public health risk posed by livestock effluents.

Evaluation of sample handling effects on serum vitamin e and cholesterol concentrations in alpacas.

Clinical cases of vitamin E deficiencies have been diagnosed in camelids and may indicate that these species are more sensitive to inadequate vitamin E in hay-based diets compared to other ruminant and equine species. In bovine, cholesterol has been reported to affect vitamin E concentrations. In order to evaluate vitamin E deficiencies in camelids, the effects of collection and storage of the blood samples prior to processing were necessary. Reports vary as to factors affecting vitamin E and cholesterol in blood samples, and diagnostic laboratories vary in instructions regarding sample handling. Blood was collected from healthy alpacas and processed under conditions including exposure to fluorescent light, serum and red blood cell contact, tube stopper contact, temperature, and hemolysis. Serum vitamin E and cholesterol concentrations were then measured. Statistical analyses found that the vitamin E concentrations decreased with prolonged contact with the tube stopper and with increasing hemolysis. Vitamin E concentration variations were seen with other factors but were not significant. Time prior to serum separation and individual animal variation was found to alter cholesterol concentrations within the sample, yet this finding was clinically unremarkable. No correlation was seen between vitamin E and cholesterol concentration, possibly due to lack of variation of cholesterol.