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Ketosis, evidenced by hyperketonemia with elevated blood β-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., (© 2024 The Author(s). Journal of Animal Breeding and Genetics published by John Wiley & Sons Ltd.)

A multicenter observational study was conducted on early lactation Holstein cows (n = 261) from 32 herds from 3 regions (Australia, AU; California, CA; and Canada, CAN) to characterize their risk of acidosis into 3 groups (high, medium, or low) using a discriminant analysis model previously developed. Diets ranged from pasture supplemented with concentrates to total mixed ration (nonfiber carbohydrates = 17 to 47 and neutral detergent fiber = 27 to 58% of dry matter). Rumen fluid samples were collected <3 h after feeding and analyzed for pH, and ammonia, d- and l-lactate, and volatile fatty acid (VFA) concentrations. Eigenvectors were produced using cluster and discriminant analysis from a combination of rumen pH, and ammonia, d-lactate, and individual VFA concentrations and were used to calculate the probability of the risk of ruminal acidosis based on proximity to the centroid of 3 clusters. Bacterial 16S ribosomal DNA sequence data were analyzed to characterize bacteria. Individual cow milk volume, fat, protein, and somatic cell count values were obtained from the closest herd test to the rumen sampling date (median = 1 d before rumen sampling). Mixed model analyses were performed on the markers of rumen fermentation, production characteristics, and the probability of acidosis. A total of 26.1% of the cows were classified as high risk for acidosis, 26.8% as medium risk, and 47.1% as low risk. Acidosis risk differed among regions with AU (37.2%) and CA (39.2%) having similar prevalence of high-risk cows and CAN only 5.2%. The high-risk group had rumen phyla, fermentation, and production characteristics consistent with a model of acidosis that reflected a rapid rate of carbohydrate fermentation. Namely, acetate to propionate ratio (1.98 ± 0.11), concentrations of valerate (2.93 ± 0.14 mM), milk fat to protein ratio (1.11 ± 0.047), and a positive association with abundance of phylum Firmicutes. The medium-risk group contains cows that may be inappetant or that had not eaten recently or were in recovery from acidosis. The low-risk group may represent cattle that are well fed with a stable rumen and a slower rumen fermentation of carbohydrates. The high risk for acidosis group had lower diversity of bacteria than the other groups, whereas CAN had a greater diversity than AU and CA. Rumen fermentation profile, abundance of ruminal bacterial phyla, and production characteristics of early lactation dairy cattle from 3 regions were successfully categorized in 3 different acidosis risk states, with characteristics differing between acidosis risk groups. The prevalence of acidosis risk also differed between regions., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Data were obtained from studies in Australia, Canada, and the United States using individual cow data from 28,230 Holstein cows to evaluate associations between parity and disease. Our goal was to develop understanding of disease risks for cows of differing parity. We hypothesized that there would be increased risks of disease and changes in metabolite concentrations with increased parity. Parity ≥5 represented 2,533 cows or 9.0%, parity 4 was 9.8% (2,778), parity 3 as 19.0% (5,355), parity 2 as 28.1% (7,925), and parity 1 was 34.1% (9,639) of the sample. Of these cows, 15.5% were in Australia, 14.7% in Canada, and 69.8% in the United States. Lactational incidence (LI) risk of clinical hypocalcemia increased with parity from 0.1% for parity 1 to 13% for parity ≥5 cows. The marked increase suggests profound differences in metabolism with increased parity. The LI of clinical mastitis was 17.4%. The odds of mastitis increased with parity to 2.5 times greater in parity ≥5 than in parity 1. The LI of lameness increased with parity; specifically, the odds of lameness was 5.6 times greater for parity ≥5 than parity 1. Dystocia incidence was 8.7% and greatest for parity 1 cows. The LI of retained placenta was 7.4% and increased with parity, with the odds for parity ≥5 2.3 times greater than for parity 1. The LI of metritis was 10% and of endometritis 14%, with the greatest odds in parity 1. The LI of clinical ketosis was 3.3% with a marked increase in odds with parity. The prevalence of subclinical ketosis was 26.8% with only cows in parity 1 having lower odds than other parities. Parity ≥5 cows had greater odds (odds ratio = 1.7) of respiratory disease than parity 1 cows, which were lesser than other parities. Metabolite concentrations were evaluated in 5,154 Holstein cows in the precalving, calving, and immediate postcalving data sets. Metabolic measures near peak lactation provided 1,906 observations. Concentrations of β-hydroxybutyrate (BHB) and nonesterified fatty acids increased with parity on d 1 to 3 of lactation and at peak lactation. On d 1 to 3 after calving differences in glucose, nonesterified fatty acids, and BHB indicated a greater reliance on mobilized lipid to export energy to peripheral tissues as BHB for greater parity cows. Differences in concentrations among parity groups were marked at times, for example >0.20 mM in Ca for parity 1 and 2 to parity ≥5 and >0.33 mM for all older parities compared with parity 1 for P on the day of calving. The marked increase suggests profound differences in metabolism with increased parity are probably influenced, in part, by increased production. We found marked differences in concentrations of metabolites with parity that are consistent with reduced reproduction, health, and body condition for higher parity cows. These unfavorable differences in metabolism in Ca, P, glucose, and cholesterol concentrations for higher parity cows also complement the often-substantial differences in disease risk with parity and suggest a need to carefully consider the parity structure in study design. Managers and advisors will need to consider methods to reduce risk of health disorders tailored to cows of different ages., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

A systematic review was conducted to assess the cost of ketosis in dairy cattle, and to elucidate how ketosis cost is estimated in each of the studies. Scientific papers addressing the economic impact of ketosis in dairy cows were identified through a search in 4 databases (Medline, ISI Web of Science, CAB Abstracts, and Agricola). The literature search was conducted with no restrictions on the date of study publication, publication type, or language. The methodological quality of the studies was assessed regarding study design, data collection, and analysis and interpretation of the study results. Of 531 identified records, 10 were selected, of which 9 were published from 2015 onward. Of the 10 studies reviewed, 9 report cost of a case of ketosis, and the estimates vary widely, with values ranging from €19 to €812. Two studies report ketosis cost at a farm level (€3.6-€29/cow per year). Among the studies, we observed great variation not only in the estimation models and inputs used (costs and losses associated with the disease) but also in the definition of ketosis and its prevalence or incidence figures. Moreover, the cost of ketosis was estimated for dairy farms in the United States, Canada, the Netherlands, Denmark, France, Germany, Spain, Sweden, Norway, and India. Consequently, there was great heterogeneity regarding herd characteristics, milk production, milk prices, culled cows' value, feed prices, and costs of veterinary services. Ketosis cost estimates vary as a consequence of all these aspects. Therefore, although most of the studies were well-designed and used high-quality data, the systematic approach review does not allow combination of the cost estimates of into a single figure. In conclusion, our review highlights an overall considerable economic impact of ketosis in dairy cattle. Economic prevention and mitigation strategies should be taken according to herd- and country-specific conditions. Ketosis cost figures reported in economic studies should always be considered carefully and interpreted with appropriate consideration of the inputs of the estimation, country context, and herd parameters., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Ketosis is one of the most frequent metabolic diseases in high-yielding dairy cows and is characterized by high concentrations of ketone bodies in blood, urine, and milk, causing high economic losses. The search for polymorphic genes, whose alleles have different effects on resistance to developing the disease, is of extreme importance to help select less susceptible animals. The aims of this study were to identify genomic regions associated with clinical and subclinical ketosis (β-hydroxybutyrate concentration) in North American Holstein dairy cattle and to investigate these regions to identify candidate genes and metabolic pathways associated with these traits. To achieve this, a GWAS was performed for 4 traits: clinical ketosis lactation 1, clinical ketosis lactation 2 to 5, subclinical ketosis lactation 1, and subclinical ketosis lactation 2 to 5. The estimated breeding values from 77,277 cows and 7,704 bulls were deregressed and used as pseudophenotypes in the GWAS. The top-20 genomic regions explaining the largest proportion of the genetic variance were investigated for putative genes associated with the traits through functional analyses. Regions of interest were identified on chromosomes 2, 5, and 6 for clinical ketosis lactation 1; 3, 6, and 7 for clinical ketosis lactation 2 to 5; 1, 2, and 12 for subclinical ketosis lactation 1; and 20, 11, and 25 for subclinical ketosis lactation 2 to 5. The highlighted genes potentially related to clinical and subclinical ketosis included ACAT2 and IGF1. Enrichment analysis of the list of candidate genes for clinical and subclinical ketosis showed molecular functions and biological processes involved in fatty acid metabolism, lipid metabolism, and inflammatory response in dairy cattle. Several genomic regions and SNPs related to susceptibility to ketosis in dairy cattle that were previously described in other studies were confirmed. The novel genomic regions identified in this study aid to characterize the most important genes and pathways that explain the susceptibility to clinical and subclinical ketosis in dairy cattle., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

Many cows culled from dairy farms arrive at livestock markets or abattoirs in compromised condition, but it is unknown whether cows are compromised at the time of culling or deteriorate during the marketing process. This study followed cull cows from farm to abattoir, monitored any deterioration, and tested the effect of providing such information on subsequent culling. For 11 mo, data were collected from 20 dairy farms, livestock markets, and abattoirs. Cows' body condition, lameness, and udder condition (as binary outcomes) were assessed at 4 points: when placed on the culling list, before leaving the farm, at auction, and at the abattoir. Based on the first 5 mo of data (baseline period), information materials were provided to 10 randomly selected farms (Informed treatment), and the other 10 served as a Control. Logistic mixed models were used to test differences in cows' condition at the 4 points of assessment, using the assessment before leaving the farm as a reference value and cow-within-farm as a random effect. The effects of providing information on outcome measures (proportion thin, severely lame, with udder edema, poor fitness for transport, euthanized, and died) were tested with a general linear model. As demand for milk increased during the baseline period, additional linear mixed models were fitted to test whether the same outcome measures were affected by time (baseline or treatment period) and treatment (Informed or Control) using farm as a random effect. While cows were on the culling list (before shipping) the odds of being thin or having poor fitness for transport increased. After leaving the farms, cows spent (mean ± standard deviation) 82 ± 46 h in the marketing system before being slaughtered. Shipping from farm to abattoir increased the odds of being thin, having udder edema, or showing poor fitness for transport [odds ratio (OR) 5.8, 95% confidence interval 4.2 to 8.1; OR 14.7, 10.7 to 20.2; and OR 7.3, 5.7 to 9.5, respectively]. Providing this information to farmers (Informed treatment) did not affect the outcome measures. Compared with the baseline, during the treatment period fewer cows were shipped when thin (4.9 versus 9.0%) and with poor fitness for transport (15.7 versus 25.8%), and fewer cows died (5.4 versus 11.9%) or were euthanized (13.3 versus 20.5%). Increased demand for milk appeared to lead to delayed culling, such that more compromised cows were sent to slaughter. The results of this study show a need for clear and specific guidelines to assess fitness for transport of cull dairy cows., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Background: Thoracic ultrasonography (US) and bronchoalveolar lavage fluid (BALF) analysis are antemortem methods used to identify the lung lesions associated with bovine respiratory disease (BRD). Accuracy of US and the cell distributions in BALF have not been characterized in calves with subclinical disease., Objectives: To evaluate the accuracy of US and BALF and describe BALF characteristics in calves with subclinical lung lesions., Animals: Twenty-five Holstein calves, 1-12 weeks old., Methods: In this prospective study, calves with low respiratory scores underwent US, BALF and postmortem examination (normal US, n = 5; comet-tails, n = 5; consolidation, n = 15). Bronchoalveolar lavage fluid was collected and analyzed for total and differential cell counts. Lung lesions were assessed by gross and histopathologic examination. Data were analyzed using nonparametric methods and relative risk analysis. The accuracy of US and BALF were estimated relative to postmortem examination., Results: The sensitivity and specificity of US for detecting lung lesions was 94% (95% CI, 69-100%) and 100% (95% CI, 64-100%), respectively. A cut-point of ≥4% BALF neutrophils was associated with the highest BALF sensitivity and specificity, 81% (95% CI, 56-94%) and 75% (95% CI, 36-95%). The presence of consolidation on US increased the risk of having a BALF neutrophil proportion ≥4% (RR, 3.9; 95% CI, 1.13-13.45; P = .003)., Conclusions and Clinical Importance: Ultrasonography accurately detects lung lesions in calves with subclinical disease. Clinicians should use a cut-point of ≥4% BALF neutrophils to diagnose subclinical respiratory disease., (Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of American College of Veterinary Internal Medicine.)

Early detection of dairy animal health issues allows the producer or veterinarian to intervene before the animals' production levels, or even survival, is threatened. An increased concentration of β-hydroxybutyrate (βHBA) is a key biomarker for diagnosis of subclinical ketosis (SCK), and provides information on the health stress in cows well before any external symptoms are observable. In this study, quantum dots (QDs) modified with cofactor nicotinamide adenine dinucleotide (NAD(+)) were prepared for the sensing event, by which the βHBA concentration in the cow's blood and milk samples was determined via fluorescence analysis of the functionalized QDs. The detection was performed on a custom designed microfluidic platform combining with a low cost and miniaturized optical sensor. The sensing mechanism was first validated by a microplate reader method and then applied to the microfluidic platform. Standard βHBA solution, βHBA in blood and milk samples from cows were successfully measured by this novel technology with a detection limit at a level of 35 µM. Side by side comparison of the developed microfluidic biosensor with a commercial kit presented its good performance., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: Determination of β-hydroxybutyrate (βHBA) is a gold standard for diagnosis of Subclinical Ketosis (SCK), a common disease in dairy cows that causes significant economic loss. Early detection of SCK can help reduce the risk of the disease progressing into clinical stage, thus minimizing economic losses on dairy cattle. Conventional laboratory methods are time consuming and labor-intensive, requiring expensive and bulky equipment. Development of portable and robust devices for rapid on-site SCK diagnosis is an effective way to prevent and control ketosis and can significantly aid in the management of dairy animal health. Microfluidic technology provides a rapid, cost-effective way to develop handheld devices for on-farm detection of sub-clinical ketosis. In this study, a highly sensitive microfluidics-based biosensor for on-site SCK diagnosis has been developed., Results: A rapid, low-cost microfluidic biosensor with high sensitivity and specificity was developed for SCK diagnosis. Determination of βHBA was employed as the indicator in the diagnosis of SCK. On-chip detection using miniaturized and cost-effective optical sensor can be finished in 1 minute with a detection limit of 0.05 mM concentration. Developed microfluidic biosensor was successfully tested with the serum samples from dairy cows affected by SCK. The results of the developed biosensor agreed well with two other laboratory methods. The biosensor was characterized by high sensitivity and specificity towards βHBA with a detection limit of 0.05 mM., Conclusions: The developed microfluidic biosensor provides a promising prototype for a cost-effective handheld meter for on-site SCK diagnosis. By using microfluidic method, the detection time is significantly decreased compared to other laboratory methods. Here, we demonstrate a field-deployable device to precisely identify and measure subclinical ketosis by specific labeling and quantification of β-hydroxybutyate in cow blood samples. A real-time on-site detection system will maximize convenience for the farmers.

The objective of this study was to assess the effect of storage conditions on the accuracy of a milk test strip for ketosis. Storage at 21 degrees C for up to 18 wk had little effect on accuracy for diagnosis and classification of subclinical ketosis.

Objective: To identify serum biochemical and hematologic variables, as measured in the week before parturition, that predict postpartum retention of the placenta (RP) in dairy cows., Design: Retrospective cohort study., Animals: 1,038 cows in 20 commercial dairy herds., Procedures: Serum concentrations of fatty acids (FAs), beta-hydroxybutyrate, cholesterol, glucose, urea, and calcium and blood leukocyte, neutrophil, lymphocyte, monocyte, and eosinophil counts were determined. These variables were evaluated for an association with development of RP by use of a multivariate logistic regression model. Parity, season of parturition, existence of twins or dystocia, body condition score, and vitamin E treatment were included in the model as covariates., Results: High serum concentrations of cholesterol and FAs were associated with an increased odds of RP. There was a 5% relative increase in the odds of RP for each 0.1 mmol/L increase in cholesterol or FAs concentration in the week before parturition. Season of parturition and twinning were also identified as risk factors., Conclusions and Clinical Relevance: These associations indicated that prepartum energy metabolism contributes to the development of RP. Serum concentrations of cholesterol and FAs may be useful to identify cows with a metabolic abnormality or energy imbalance that might predispose them to RP and should be interpreted in conjunction with clinical risk factors such as twinning, dystocia, or parturient paresis.

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne's disease in cattle and other farm ruminants, and is also a suspected pathogen of Crohn's disease in humans. Development of diagnostic methods for MAP infection has been a challenge over the last few decades. The objective of this study was to investigate the relationship between different methods for detection of MAP in milk and fecal samples. A total of 134 milk samples and 110 feces samples were collected from 146 individual cows in 14 MAP-infected herds in southwestern Ontario. Culture, IS900 polymerase chain reaction (PCR) and nested PCR methods were used for detecting MAP in milk; results were compared with those of fecal culture. A significant relationship was found between milk culture, direct PCR, and nested PCR (P < 0.05). The fecal culture results were not related to any of the 3 assay methods used for the milk samples (P > 0.10). Although fecal culture showed a higher sensitivity than the milk culture method, the difference was not significant (P = 0.2473). The number of MAP colony-forming units (CFU) isolated by culture from fecal samples was, on average, higher than that isolated from milk samples (P = 0.0083). There was no significant correlation between the number of CFU cultured from milk and from feces (Pearson correlation coefficient = 0.1957, N = 63, P = 0.1243). The animals with high numbers of CFU in milk culture may not be detected by fecal culture at all, and vise versa. A significant proportion (29% to 41%) of the positive animals would be missed if only 1 culture method, instead of both milk and feces, were to be used for diagnosis. This suggests that the shedding of MAP in feces and milk is not synchronized. Most of the infected cows were low-level shedders. The proportion of low-level shedders may even be underestimated because MAP is killed during decontamination, thus reducing the chance of detection. Therefore, to identify suspected Johne's-infected animals using the tests in this study, both milk and feces samples should be collected in duplicate to enhance the diagnostic rate. The high MAP kill rate identified in the culture methods during decontamination may be compensated for by using the nested PCR method, which had a higher sensitivity than the IS900 PCR method used.

To identify management practices associated with an increased within-herd prevalence of Cryptosporidium parvum shedding on dairy farms in southern Ontario, fecal samples were taken from 1089 calves aged 7-28 days, from 119 herds. Information on management practices was obtained by administering a questionnaire compiled using a modified Delphi technique. Data were analyzed using univariable and multivariable negative binomial regression. Overall, 30% of the calves in the study were shedding C. parvum oocysts, with at least one positive calf detected in 77% of herds. Within-herd prevalence ranged from 0 to 80%. Predictors significantly associated with an increased prevalence of shedding in multivariable modelling were the use of calf scour prophylaxis in cows (risk ratio [RR] 1.70, P<0.01) and calves (RR 1.38, P=0.02) and the feeding of milk replacer in the first week of life (RR 1.40, P=0.02). In contrast, the presence of concrete flooring in calf housing areas (RR 0.59, P<0.01) and the use of soap or detergent when washing calf feeding utensils (RR 0.61, P<0.01) appeared to be protective.

This work was conducted to investigate calf-level factors that influence the risk of neonatal diarrhea and shedding of Cryptosporidium parvum oocysts in calves, on dairy farms in Ontario with histories of calf diarrhea or cryptosporidiosis. Fecal samples were collected weekly for 4 weeks from each of 1045 calves under 30 days of age on 11 dairy farms in south-western Ontario during the summer of 2003 and the winter of 2004. A questionnaire designed to gather information on calf-level management factors was administered on farm for each calf in the study. Samples were examined for C. parvum oocysts by microscopy, and a subset of specimens was also tested for enterotoxigenic Escherichia coli, Salmonella, bovine rotavirus and bovine coronavirus. The consistency of each sample was scored and recorded at the time of collection in order to assess the presence or absence of diarrhea. In addition, a blood sample was taken from each calf upon enrollment in the study, for assessment of maternal antibody transfer and for polymerase chain reaction testing for persistent bovine viral diarrhea virus infection. Using the GLLAMM function in Stata 9.0, multilevel regression techniques were employed to investigate associations between management practices and the risk of C. parvum shedding or diarrhea. C. parvum oocysts were detected in the feces of 78% of the 919 calves from which all four fecal samples had been collected. Furthermore, 73% of the 846 calves for which all four fecal consistency scores had been recorded were diarrheic at the time of collection of at least one sample. Significant predictors of the calf-level risk of C. parvum shedding included the use of calf diarrhea prophylaxis in pregnant cows, and the type of maternity facilities in which the calves were born. Factors associated with an increased risk of diarrhea were leaving the calf with the dam for more than an hour after birth, and the birth of a calf in the summer as opposed to winter. Calves shedding C. parvum oocysts had 5.3 (95% CI 4.4, 6.4) times the odds of diarrhea than non-shedding calves, controlling for other factors included in the final multivariable model. Furthermore, infected calves shedding more than 2.2 x 10(5) oocysts per gram of feces were more likely to scour than infected calves shedding lower numbers of oocysts (OR= 6.1, 95% CI 4.8, 7.8). The odds of diarrhea in calves shedding oocysts that had been allowed to remain with their dams for more than an hour were higher than the odds of diarrhea in shedding calves that had been separated from their dams within an hour after birth.

Clostridium difficile toxins were associated with calf diarrhea in a recent retrospective study; however, no causal relationship has been prospectively investigated. This infection study tested whether the oral inoculation of neonatal calves with a toxigenic strain of C. difficile (PCR-ribotype 077) results in enteric disease. Fourteen 6-24 h old male colostrums-fed Holstein calves, received either three doses of C. difficile (1.4 x 10(8) +/- 3.5 x 10(8) cfu) (n = 8) or sterile culture broth (n = 6). Calves were euthanized on day 6 or after the onset of diarrhea, whichever came first. Fecal and intestinal samples were blindly cultured for C. difficile, and tested for its toxin A/B (C. difficile TOX A/B II ELISA, Techlab). PCR-ribotyping was used to compare inoculated and recovered isolates. Diarrhea was observed in all control calves and 3/8 of inoculated calves (p = 0.03), but it did not occur in calves that tested positive for C. difficile toxins. Fecal toxins were identified only from two controls. PCR-ribotyping confirmed the presence of C. difficile PCR-ribotype 077 in samples of all inoculated calves, but not from controls. The identification of five other PCR-ribotypes in 3/8 (37.5%) and 2/6 (33.3%) of inoculated and control calves, respectively, indicated early natural infection (< or = 24h of age). Five of 14 cecal samples had C. difficile (p = 0.01). In conclusion, the oral administration of C. difficile PCR-ribotype 077 to neonatal calves resulted in fecal/intestinal colonization but not in detection of toxins, or signs of enteric disease. Further studies are required to investigate the clinical relevance of C. difficile in calves.

This study randomly surveyed 50 dairy herds in Ontario; 18% and 30% of herds had 2 or more milk or serum enzyme-linked immunosorbent assay (ELISA)-positive cows, respectively. The apparent cow level prevalence was 1.7% and 2.6% on the milk and serum ELISA, respectively. The serum and milk assays agreed moderately.

There is a need for simple and inexpensive diagnostic and screening tests for the detection of Cryptosporidium parvum infection in calves. A sucrose wet mount test and a lateral immunochromatography test were evaluated for epidemiological sensitivity and specificity, cost per test, simplicity, test time and ease of batching. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the Cryptosporidium oocyst wall protein (COWP) gene locus, with gel electrophoresis, was used as a gold standard. Cohen's kappa statistic of agreement (kappa) between the Ontario Veterinary College (OVC) sucrose wet mount test and COWP PCR-RFLP was 0.82, and the sensitivity and specificity of the OVC sucrose wet mount test were 88.6% and 93.8%, respectively. The sensitivity and specificity of the lateral immunochromatography test were 78.3% and 93.3%, respectively, and agreement between this test and PCR-RFLP was good (kappa=0.73). There was substantial agreement between the OVC sucrose wet mount test and the lateral immunochromatography test (kappa=0.84). Both tests were inexpensive and easy to use; however, the lateral immunochromatography test was faster and simpler to perform than the sucrose wet mount test, and was generally more user-friendly. These tests provide practitioners and researchers with cheap, quick and accurate methods of detecting C. parvum infection in young calves.

A total of 1317 Holstein cows from 45 farms in the Canadian provinces of Quebec, Prince Edward Island (PEI) and Ontario were enrolled in a randomized trial during 1998 and 1999 to further confirm the efficacy of a monensin controlled release capsule in preventing periparturient disease in lactating dairy cows. Cows were randomized on the farms to receive either a monensin controlled release capsule (CRC) 2 to 4 wk before expected calving or to serve as negative controls. Health data were collected for 90 d postcalving and were analyzed with logistic regression accounting for the intraherd correlation with generalized estimating equations. Monensin CRC significantly reduced the incidence of both clinical ketosis and abomasal displacement post-calving. There was a numerical but nonsignificant decrease in the incidence of retained placenta in cows receiving a monensin CRC. A pooled analysis of two separate but similar studies (conducted in 1995 and 1998) demonstrated a strengthened association between monensin CRC administration precalving and reduced periparturient disease. A 40% reduction in both abomasal displacement and clinical ketosis was observed with precalving administration of a monensin CRC. In addition, the larger dataset highlighted a trend for a 25% reduction in the incidence of retained placenta in monensin-treated cows. Improved energy metabolism as a result of monensin treatment is likely the mechanism for the reduction in incidence of all three of these diseases. Thus the term "energy associated disease" was created to assess the combined impact of the precalving monensin treatment on the incidence of retained placenta, displaced abomasum, and clinical ketosis. The monensin controlled release capsule reduced the incidence of energy associated disease by 30%.

Results of studies on metabolic variables that exist prior to the diagnosis of displaced abomasum (DA) indicate that subclinical ketosis is a risk factor for DA. Serum aspartate aminotransferase activity, serum and milk betahydroxybutyric acid concentrations, and milk fat-to-protein ratios were often elevated before the diagnosis of DA, and these variables may be used to predict DA. Prevention of subclinical ketosis may prevent DA.

Subclinical ketosis is an important and common condition of early-lactation dairy cattle. It is associated with losses in milk production and increased risk of periparturient disease. Prevention depends on several factors, including proper transition-cow nutrition, management of body condition, and the use of certain feed additives such as niacin, propylene glycol, and ionophores. All currently available cowside tests for subclinical ketosis have certain limitations in their use. Effective monitoring schemes for subclinical ketosis can be developed, however, and these may be useful in many herd health programs.

First Dairy Herd Improvement Corporation (DHI) test milk that was sampled prior to displaced abomasum (DA) diagnosis was used to examine the association between milk yield (kg), fat (%), protein (%), or fat/protein ratio and DA in dairy cows. 27 DA cases were matched to 3 controls per case by herd and calving date. Logistic regression was used to examine the effect of each milk parameter on the odds of DA, adjusting for lactation number and days in milk (DIM) at test. Higher odds of DA diagnosis was found with lower milk yield, higher milk fat percentage, lower milk protein percentage and higher fat/protein ratio. We conclude that milk yield, fat percentage, protein percentage and fat/protein ratio may be used to detect cows at risk for DA. Our findings also indicate that cows diagnosed with DA were energy deficient prior to DA diagnosis.

The objective of this study was to evaluate the characteristics of five ketone tests for use with milk to detect subclinical ketosis [defined as > 1200 mumol of beta-hydroxybutyrate (BHBA)/L of blood] in dairy cows. The tests studied were the Ketolac BHB strip to detect BHBA (Hoechst, Unterschleissheim, Germany) and four tests based on sodium nitroprusside to detect acetoacetate (Rothera tests) [Ketostix strip (Bayer, Etobicoke, ON, Canada), Bioketone powder (Societé d' Analyses Biopharmaceutiques, Laval, QC, Canada), Ketocheck powder (Great States, St. Joseph, MO), and Utrecht powder (University of Utrecht, Utrecht, The Netherlands)]. Milk samples (n = 529) from 266 cows in 25 Ontario dairy herds were used for this investigation. The Ketolac BHB strip at 50 and 100 mumol of BHBA/L of milk showed sensitivities of 92 and 72%, respectively. This test was more sensitive for subclinical ketosis than were any of the Rothera tests. The sensitivities of the Rothera tests were 43, 33, 28, and 5%, respectively, for Utrecht powder, Bioketone powder, Ketocheck powder, and Ketostix strip. The Rothera tests were highly specific, as was the Ketolac BHB strip at 200, 500, and 1000 mumol of BHBA/L of milk (specificity, > 97%). The prevalence of subclinical ketosis was highest during the first 6 wk of lactation, and a peak was detected during the 2nd wk of lactation. Of the tests evaluated in this study, Ketolac BHB strip was the most useful to monitor subclinical ketosis in dairy herds.

Two commercial milk ketone tests (Ketocheck) Great States, St. Joseph, MI and Ketolac BHB; Hoechst, Unterschleissheim, Germany) were used with milk samples collected within 2 wk postpartum to evaluate the use of these tests in the prediction of left displaced abomasum in dairy cows. Odds ratios, sensitivities, specificities, and likelihood ratios were determined. For the study of Ketocheck, 47 milk samples from 26 cows that were later diagnosed with left displaced abomasum were taken at a median of 6 d postpartum (12 d prior to diagnosis of the disease. For the study of Ketolac BHB, 26 milk samples from 15 cows that were later diagnosed with left displaced abomasum were taken at a median of 6 d postpartum (11 d prior to diagnosis of the disease). Each cow that was subsequently diagnosed with left displaced abomasum was matched with three control cows by herd and calving date. Positive Ketocheck results and Ketolac BHB results that indicated > or = 100 or > or = 200 mumol of beta-hydroxybutyrate (BHBA)/L of milk were significantly associated with the subsequent diagnosis of left displaced abomasum. The sensitivity of Ketocheck for the prediction of left displaced abomasum was 22%; specificity was 96%. The sensitivity of Ketolac BHB for the prediction of left displaced abomasum was 46% when the results indicated > or = 100 mumol of BHBA/L and was 34% when the results indicated > or = 200 mumol of BHBA/L. Ketocheck and Ketolac BHB might be used as tests for the prediction of left displaced abomasum when used with milk sampled within 2 wk postpartum. Hyperketolactia is a significant risk factor of left displaced abomasum.

Objective: To evaluate aspartate transaminase (AST) activity and beta-hydroxybutyrate (BHB) concentration in blood obtained during the first or second postpartum (PP) week as tests for prediction of subsequent left displaced abomasum (LDA) diagnosis in dairy cows., Animals: 36 cows with LDA tested at a mean 3 PP days, which was 7 to 22 days prior to LDA diagnosis (25, 75% quantiles), and 28 cows with LDA tested at 10 PP days, which was 5 to 18 days prior to LDA diagnosis, were matched to 3 controls per case by herd and calving date. Data were available from a large field study., Procedure: Odds ratio, sensitivity, specificity, and likelihood ratio were determined for various AST and BHB cutoff values., Results: AST, using cutoff values between 100 and 180 U/L, and BHB, using cutoff values between 1,000 and 1,600 mumol/L, were significantly associated with subsequent LDA diagnosis. When cutoff values were increased, odds ratio and likelihood ratio increased; however, sensitivity decreased and specificity increased., Conclusion and Clinical Relevance: AST activity and BHB concentration in blood obtained during the first or second PP week might be useful as predictors of subsequent LDA diagnosis.

First DHI test milk that was sampled prior to displaced abomasum (DA) diagnosis was used to evaluate milk fat/protein ratio (FPR) for prediction of subsequent DA in dairy cows. Odds ratio, sensitivity, specificity, predictive values, and likelihood ratio were determined. Twenty-seven DA cases were matched to three controls per case by herd and calving date. Milk was tested 18 to 23 days (95% CI) after calving, which was 9 to 17 days (95% CI) prior to DA diagnosis. Adjusted for parity and days in milk, a fat/protein ratio > or = 1.4 was 8.6 times more likely (95% CI, 2.8 to 26) to come from a cow subsequently diagnosed with DA than a fat/protein ratio < 1.4. Using the cut off value of 1.4, the sensitivity of the FPR for DA was 80% and the specificity was 69%. A response operating characteristics curve indicated that the minimum sum of false negative and false positive results was at an FPR cut off value of 1.4. The likelihood ratio indicated that fat/protein ratios > or = 1.6 are 3.3 times more likely to come from cows that are diagnosed subsequently with DA than from cows without DA. The fat/protein ratio in first DHI test milk might be useful as a test for subsequent DA in dairy cows.