Vaccination protocols in Québec dairy herds.

Information is needed on vaccination protocols used by veterinarians and dairy producers to prevent and control infections in dairy herds. This observational study described farm's vaccination standard operating procedures (SOP) developed by veterinarians in collaboration with dairy producers in Québec. Data pertaining to vaccination protocols and dairy producer practices were collected as part of the biosecurity component of the National Mandatory Quality Assurance Certification Program (proAction). Generalized statistical mixed-effects models were used to assess associations between dairy herd characteristics and the vaccination SOP, encompassing various vaccination types. These included any vaccination, core vaccines only (bovine respiratory syncytial virus, infectious bovine rhinotracheitis herpesvirus, parainfluenza virus type 3, bovine viral diarrhea virus type 1 and type 2) and vaccination against diarrhea, mastitis, or clostridial diseases. These models accounted for random variations related to clustering by veterinarians and veterinary clinics. Furthermore, the variance of the outcome was partitioned into producer, veterinarian, and veterinary clinic levels to explore the proportion of the total variance attributable to each group. A total of 3,759 standardized vaccination procedures completed between 2018 and 2021 were analyzed. At least one vaccination target was included in the vaccination SOP in 89% of the dairy herds. The most frequently included vaccine in the SOP was core vaccines, comprising 88%, followed by mastitis (22%), neonatal diarrhea (18%), and clostridial diseases (15%). The vaccination SOPs, particularly core, mastitis, and diarrhea vaccinations, mainly varied due to the veterinarian's characteristics, followed by the clinic's characteristics. In contrast, the decision to included clostridial vaccination primarily varied with the veterinary clinic (76%). Organic producers generally included fewer vaccinations in their SOPs, including core vaccines, than conventional producers. In addition, producers who were providing access to pasture had fewer vaccination SOP for vaccination against mastitis and neonatal diarrhea but more vaccination SOP for clostridial vaccination.

Development of a multivariable prediction model to identify dairy calves too young to be transported to auction markets in Canada using simple physical examination and body weight.

Calves born on Eastern Canadian dairy farms that are not kept in the herds are traditionally sold through auction markets and are raised for meat purposes such as veal calves. Since February 2020, a new Canadian federal regulation has forbidden calves <9 d old to be sold through auction markets. However, in the absence of a real-time birth registry consultation system, it would be of interest to look for predictors that could be associated with age to allow identification of calves too young to be transported. In the current retrospective cross-sectional study, 1,178 calves with a declared birth date (411 calves aged <9 d old; 34.9%) were assessed in 2 large Québec auction sites. Easy-to-record covariates [body weight (BW), breed phenotype, and presence of an umbilical cord remnant] as well as other clinical signs (umbilical swelling, enlargement, umbilical pain, wet umbilicus, skin tent, sunken eyes, ocular and nasal secretion, and hide cleanliness) were assessed. Two logistic regression models using age as a dichotomous dependent variable (<9 d old vs ≥9 d old) were built. The first model (model 1) considered all covariates, which were selected after univariable analyses and a backward stepwise selection process, whereas a more pragmatic model (model 2) only included the 3 easy-to-record variables (i.e., BW, breed, umbilical cord). Both models had similar accuracy to detect calves <9 d old (sensitivity of 38.4 and 37.5%, and specificity of 85.7 and 84.6% for model 1 and 2, respectively). Model 2 was subsequently more specifically studied as it employs a faster and easier assessment. Decision thresholds were tested for their robustness based on misclassification cost term (MCT) analysis with various prevalence of calves <9 d old and various costs of false-negative:false-positive ratio. Despite statistical significance, model accuracy, even if refined with MCT analysis, was limited at the individual level, showing the limits of using physical signs and BW or their combination as a reliable proxy of age. The sensitivity of these models to find calves <9 d old was not to be used for monitoring compliance with the Canadian federal regulation. The relatively high model specificity may help to use this model as a rule-in test (i.e., targeting positive calves for further investigation) rather than a rule-out test (due to its low sensitivity).

Systematic review and meta-analysis of refractometry for diagnosis of inadequate transfer of passive immunity in dairy calves: Quantifying how accuracy varies with threshold using a Bayesian approach.

Inadequate transfer of passive immunity (TPI) is associated with increased risk for calfhood disease and increased risk of mortality and morbidity. Accurately diagnosing calves and herds with inadequate TPI is of primary importance and brix (BRIX) or classical refractometer (REF) devices are more practical for this purpose than measuring the serum immunoglobulin G concentration in neonatal calves. We previously reported a systematic review and meta-analysis for quantifying the pooled accuracy of BRIX and REF for detecting calves with serum IgG < 10 g/L noting that sparse data were available especially because studies did not report the same thresholds. We updated the previous systematic review using different methods that accounted for the test results distribution in calves with or without inadequate TPI. With this approach, all reported cut-offs for a specific study are used in that Bayesian approach that quantifies how accuracy varied among all reported thresholds. Five new manuscripts were included, which represented 4 new studies since the initial study was performed. A total of 11 REF and 9 BRIX studies were available. The meta-analytic methods allowed reporting variation of the true and false positive rate across and among all reported cut-offs. Pooled points estimates (95 % Bayesian credible intervals) for sensitivity (Se) and specificity (Sp) of REF < 5.5 g/L were 86.1 % (68.5-97.9%) and 76.2 % (65.9-88.4%) whereas BRIX < 8.4 % was associated with Se of 91.6 % (77.2-99.5%) and Sp of 88.2 % (65.4-99.8%). Interestingly, the accuracy (Se + Sp-1) was generally higher for BRIX than for REF at the reported cut-offs. Besides the benefit of providing pooled estimates for all reported and unreported BRIX and REF thresholds, the general framework used in this study could potentially be used in many veterinary diagnostic tests studies that reported multiple thresholds accounting for potentially different tests distributions in population with and without the target condition.

Factors affecting dairy calf price in auction markets in Québec, Canada: 2008-2019.

Dairy calves not kept for replacement are sold at young age in Québec auction markets for white and grain-fed veal calf production. The province of Québec produces 80% of the Canadian veal meat, but little information is available on the factors associated with the calves' price per crude weight (Can$/kg; Can$1 = US$0.78 at time of writing). The characteristics of calves sold in Québec auction markets from 12 complete years (2008-2019) were retrospectively studied. The calves' weight, breed and sex, the year and season of sale, the auction site, as well as the estimated distance traveled between the farm of origin and the auction site were analyzed as potential covariates associated with calf price. Two multivariable logistic models associated with low sale value (below the 10th or the 25th percentile of the day price) and 2 models associated with good sale characteristics (above the 50th or the 75th percentile of the day price) were built. The median distance between the farm and the auction site was 52 km (interquartile range: 30-95 km). Only 5% of calves traveled distances greater than 220 km. The weight, breed, sex, and auction sites explained most of the variability in the different models. Distance traveled and multiple interactions were also significantly associated with the outcomes. Calves with body weight from 48 to <56 kg were sold in higher percentiles of the day than lighter or heavier calves. Beef-crossed calves had better sale prices than Holstein, whereas colored dairy calves had lower sale characteristics than both Holstein and beef-crossed calves. The effect of distance traveled was complex, varying depending on the model and interactions, and explained a small portion of the total deviance in every model. Calves traveling from distances ≥110 km had lower sale characteristics in summer and fall in the different studied models. This study gives relevant insights on calves' characteristics associated with good versus low sale prices in Québec auction markets.

Comparison between thoracic ultrasonography and thoracic radiography for the detection of thoracic lesions in dairy calves using a two-stage Bayesian method.

Infectious bronchopneumonia is a lower respiratory tract disease with major economic consequences in dairy calves. Thoracic radiography (TR) and thoracic ultrasonography (TUS) are two imaging diagnostic procedures available in bovine medicine for identifying thoracic lesions. However, no study has investigated whether one of these tests is superior to the other or if they provide comparable results for the detection of thoracic lesions in calves. The objective of this study was therefore to estimate and to compare the performances of TUS and TR for the detection of thoracic lesions in dairy calves. A prospective cross-sectional study was performed in a hospital setting. A total of 50 calves (≥7 days old; ≤100 kg; standing; pCO2 ≥ 53 mmHg; any reason of presentation) were enrolled. Every calf underwent TUS and TR. Only calves with thoracic lesions on TUS and/or TR were controlled by thoracic computed tomography (CT) (the gold standard). Calves without lesions were not controlled by CT. A two-stage Bayesian framework was used. The sensitivities (Se) and specificities (Sp) of both tests individually and used in series or parallel were estimated. The Se and Sp of TUS were 0.81 (95 % BCI (Bayesian Credible Interval): 0.65; 0.92) and 0.90 (95 % BCI: 0.81; 0.96), respectively. The Se and Sp of TR were 0.86 (95 % BCI: 0.62; 0.99) and 0.89 (95 % BCI: 0.67; 0.99), respectively. This study did not reveal any differences between both tests. Using TUS and TR in series was more specific than using both tests in parallel. The performances of TUS alone were not different from the performances of both tests in series or in parallel. In conclusion, TUS and TR were equivalent in detecting thoracic lesions in this study. Using TUS alone allowed an accurate detection of thoracic lesions in dairy calves. Further studies enrolling a larger sample (> 400 calves) and allowing adequate power to be achieved would be necessary to confirm these results.

Validation of serum gamma-glutamyl transferase activity and body weight information for identifying dairy calves that are too young to be transported to auction markets in Canada.

Dairy calves are at risk of being stressed when transported during the first week of life. A new Canadian federal rule will forbid transportation of calves younger than 9 d old to auction market. However, in the absence of reliable information to determine birth date, other indirect methods would be of interest. This study aimed to determine the prediction accuracy of body weight, Brix refractometry, and serum gamma-glutamyl transferase (GGT) activity for determining if a calf was not fit to be transported (i.e., <9 d old). For this purpose, we used 284 calves with a known birth date from a cross-sectional and a prospective cohort study. A logistic regression model was built based on multivariable analysis as well as a misclassification cost term analysis. Because of the collinearity between GGT activity and Brix value and lower discrimination of Brix value, the GGT activity was retained for the main model. The final logistic regression model contained body weight and log-transformed GGT activity value. The misclassifications of the logistic model was minimized using a model probability threshold ≥0.55 with a sensitivity of 70.4% and a specificity of 77.3%. This probability threshold was relatively robust for various prevalence and false negative to false positive cost ratios. The prediction accuracy of this model was moderate at the individual level, but is helpful in calves with a reasonable suspicion of being less than 9 d old.

Accuracy of paratuberculosis diagnostic tests in small ruminants: protocol for a systematic review and meta-analysis.

Paratuberculosis is a worldwide infectious disease caused by Mycobacterium avium ssp. paratuberculosis (MAP). Various ruminant species can be affected by the disease, and the diagnosis of the disease is challenging in the absence of a gold standard test. The aim of this systematic review protocol is to determine the accuracy of the direct and indirect diagnostic tests for MAP infection with a special focus on sheep and goats.

Validation of a clinical scoring system for bovine respiratory disease complex diagnosis in preweaned dairy calves using a Bayesian framework.

Bovine respiratory disease complex is a major cause of illness in dairy calves. The diagnosis of active infection of the lower respiratory tract is challenging on daily basis in the absence of accurate clinical signs. Clinical scoring systems such as the Californian scoring system, are appealing but were developed without considering the imperfection of reference standard tests used for case definition. This study used a Bayesian latent class model to update Californian prediction rules. The results of clinical examination and ultrasound findings of 608 preweaned dairy calves were used. A model accounting for imperfect accuracy of thoracic ultrasound examination was used to obtain updated weights for the clinical signs included in the Californian scoring system. There were 20 points (95% Bayesian credible intervals: 11-29) for abnormal breathing pattern, 16 points (95% BCI: 4-29) for ear drop/head tilt, 16 points (95% BCI: 9-25) for cough, 10 points (95% BCI: 3-18) for the presence of nasal discharge, 7 points (95% BCI: -1 to 8) for rectal temperature ≥39.2 °C, and -1 points (95% BCI: -9 to 8) for the presence of ocular discharge. The optimal cut-offs were determined using the misclassification cost-term term (MCT) approach with different possible scenarios of expected prevalence and different plausible ratio of false negative costs/false positive costs. The predicted probabilities of active infection of the lower respiratory tract were also obtained using posterior densities of the main logistic regression model. Depending on the context, cut-off varying from 9 to 16 can minimized the MCT. The optimal cut-off decreased when expected prevalence of disease and false negative/false positive ratio increased.

Non-invasive acquisition of fetal ECG from the maternal xyphoid process: a feasibility study in pregnant sheep and a call for open data sets.

OBJECTIVE: The utility of fetal heart rate (FHR) monitoring can only be achieved with an acquisition sampling rate that preserves the underlying physiological information on the millisecond time scale (1000 Hz rather than 4 Hz). For such acquisition, fetal ECG (fECG) is required, rather than the ultrasound to derive FHR. We tested one recently developed algorithm, SAVER, and two widely applied algorithms to extract fECG from a single-channel maternal ECG signal recorded over the xyphoid process rather than the routine abdominal signal. APPROACH: At 126dG, ECG was attached to near-term ewe and fetal shoulders, manubrium and xyphoid processes (n = 12). fECG served as the ground-truth to which the fetal ECG signal extracted from the simultaneously-acquired maternal ECG was compared. All fetuses were in good health during surgery (pH 7.29 ± 0.03, pO2 33.2 ± 8.4, pCO2 56.0 ± 7.8, O2Sat 78.3 ± 7.6, lactate 2.8 ± 0.6, BE -0.3 ± 2.4). MAIN RESULT: In all animals, single lead fECG extraction algorithm could not extract fECG from the maternal ECG signal over the xyphoid process with the F1 less than 50%. SIGNIFICANCE: The applied fECG extraction algorithms might be unsuitable for the maternal ECG signal over the xyphoid process, or the latter does not contain strong enough fECG signal, although the lead is near the mother's abdomen. Fetal sheep model is widely used to mimic various fetal conditions, yet ECG recordings in a public data set form are not available to test the predictive ability of fECG and FHR. We are making this data set openly available to other researchers to foster non-invasive fECG acquisition in this animal model.

Systematic Review and Meta-Analysis of Diagnostic Accuracy of Serum Refractometry and Brix Refractometry for the Diagnosis of Inadequate Transfer of Passive Immunity in Calves.

BACKGROUND: Transfer of passive immunity in calves can be assessed by direct measurement of immunoglobulin G (IgG) by methods such as radial immunodiffusion (RID) or turbidimetric immunoassay (TIA). IgG can also be measured indirectly by methods such as serum refractometry (REF) or Brix refractometry (BRIX). OBJECTIVES: To determine the accuracy of REF and BRIX for assessment of inadequate transfer of passive immunity (ITPI) in calves. DESIGN: Systematic review and meta-analysis of diagnostic accuracy studies. METHODS: Databases (PubMed and CAB Abstract, Searchable Proceedings of Animal Science) and Google Scholar were searched for relevant studies. Studies were eligible if the accuracy (sensitivity and specificity) of REF or BRIX was determined using direct measurement of IgG by RID or turbidimetry as the reference standard. The study population included calves <14 days old that were fed with natural colostrum (colostrum replacement products were excluded). Quality assessment was performed by the QUADAS-2 tool. Hierarchical models were used for meta-analysis. RESULTS: From 1,291 references identified, 13 studies of 3,788 calves were included. Of these, 11 studies evaluated REF and 5 studies evaluated BRIX. The median (range) prevalence of ITPI (defined as calves with IgG <10 g/L by RID or TIA) was 21% (1.3-56%). Risk of bias and applicability concerns were generally low or unclear. For REF, summary estimates were obtained for 2 different cutoffs: 5.2 g/dL (6 studies) and 5.5 g/dL (5 studies). For the 5.2 g/dL cutoff, the summary sensitivity (95% CI) and specificity (95% CI) were 76.1% (63.8-85.2%) and 89.3% (82.3-93.7%), and 88.2% (80.2-93.3%) and 77.9% (74.5-81.0%) for the 5.5 g/dL cutoff. Due to the low number of studies using the same cutoffs, summary estimates could not be obtained for BRIX. CONCLUSIONS AND CLINICAL IMPORTANCE: Despite their widespread use on dairy farms, evidence about the optimal strategy for using refractometry, including the optimal cutoff, are sparse (especially for BRIX). When using REF to rule out ITPI in herds, the 5.5 g/dL cutoff may be used whereas for ruling in ITPI, the 5.2 g/dL cutoff may be used.

Incidence of fecal excretion of Mycobacterium avium subsp. paratuberculosis in dairy cows before and after the enrolment in the Québec voluntary program.

Paratuberculosis is a chronic and contagious enteric disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). This disease of worldwide distribution is responsible for significant economic losses and the bacteria itself has been linked to human Crohn's disease. Paratuberculosis control programs focus on reducing MAP transmission by implementing better management practices that target infection routes. In Québec, a Voluntary Paratuberculosis Prevention and Control Program (QVPPCP) was launched in 2007. The objectives of this prospective cohort study were threefold. The first was to describe the changes in the incidence of fecal excretion of MAP in cows born before and after farm enrolment in the QVPPCP. The second was to estimate the impact of the risk of within-herd transmission of MAP (measured by the risk assessment score (RAS)) on the incidence of fecal excretion of MAP. And the third was to evaluate the impact of calf rearing practices on the incidence of fecal excretion of MAP. Eighteen MAP-positive herds were visited annually from 2011 to 2015. At each visit, individual fecal samples from all adult cows were collected. MAP was cultured using liquid media and an automated system. A risk assessment questionnaire was completed upon enrolment in the QVPPCP and at each visit. The RAS of the farm was attributed to each cow according to its birthdate. Cox proportional hazards models were used to estimate the hazard ratios (HR) for the exposure variables. Herd clustering was taken into account using robust standard errors. A total of 2158 cows were included (cohort born before n=919; cohort born after n=1239). The incidence and hazard of fecal excretion were significantly lower for the cohort-after than the cohort-before (incidence rate ratio=0.38; 95% CI: 0.18-0.78 and HR=0.48; 95% CI: 0.23-0.98). The HR of fecal excretion for cows exposed to a high RAS was 2.20 times (95% CI: 1.21-3.99) that of cows exposed to a low RAS. Poor calving cow hygiene (HR=3.41; 95% CI: 1.40-8.31) and contact between pre-weaned heifers and adult cows or their feces were significantly associated with an increased hazard of fecal excretion of MAP (HR=2.66; 95% CI: 1.08-6.56). Our results suggest that enrolment in the QVPPCP reduces the risk of MAP fecal excretion. They support the hypothesis that contact between calves and adult cows or their feces increases MAP transmission. The incidence results also suggest that MAP prevalence could be reduced to low levels regardless of initial MAP prevalence.

α7 nicotinic acetylcholine receptor signaling modulates the inflammatory phenotype of fetal brain microglia: first evidence of interference by iron homeostasis.

Neuroinflammation in utero may result in life-long neurological disabilities. Microglia play a pivotal role, but the mechanisms are poorly understood. No early postnatal treatment strategies exist to enhance neuroprotective potential of microglia. We hypothesized that agonism on α7 nicotinic acetylcholine receptor (α7nAChR) in fetal microglia will augment their neuroprotective transcriptome profile, while the antagonistic stimulation of α7nAChR will achieve the opposite. Using an in vivo - in vitro model of developmental programming of neuroinflammation induced by lipopolysaccharide (LPS), we validated this hypothesis in primary fetal sheep microglia cultures re-exposed to LPS in presence of a selective α7nAChR agonist or antagonist. Our RNAseq and protein level findings show that a pro-inflammatory microglial phenotype acquired in vitro by LPS stimulation is reversed with α7nAChR agonistic stimulation. Conversely, antagonistic α7nAChR stimulation potentiates the pro-inflammatory microglial phenotype. Surprisingly, under conditions of LPS double-hit an interference of a postulated α7nAChR - ferroportin signaling pathway may impede this mechanism. These results suggest a therapeutic potential of α7nAChR agonists in early re-programming of microglia in neonates exposed to in utero inflammation via an endogenous cerebral cholinergic anti-inflammatory pathway. Future studies will assess the role of interactions between inflammation-triggered microglial iron sequestering and α7nAChR signaling in neurodevelopment.

Cerebrospinal Fluid Analysis in Recumbent Adult Dairy Cows With or Without Spinal Cord Lesions.

BACKGROUND: Diagnosis of central nervous system (CNS) lesions in recumbent dairy cattle (RDC) is challenging because neurologic examination is limited and medical imaging often is challenging or unrewarding. Cerebrospinal fluid (CSF) analysis is useful in the diagnosis of CNS disorders in cattle. However, its utility in identifying spinal cord lesions in RDC remains to be evaluated. HYPOTHESIS/OBJECTIVES: We hypothesized that CSF analysis would discriminate between RDC with and without spinal cord lesions. ANIMALS: Twenty-one RDC with spinal cord lesions (RDC+) and 19 without (RDC-) were evaluated. METHODS: Spinal cord lesions were confirmed at necropsy. Signalment, clinical findings, and CSF results were compared retrospectively. Total nucleated cell count and differential, protein concentration, and red blood cell count in RDC+ and RDC- were compared. RESULTS: Neoplasia, trauma, and infectious processes were the most frequent spinal cord lesions identified. Cerebrospinal fluid protein concentrations and TNCC were significantly higher in RDC+ compared to RDC- (P = .0092 and P = .0103, respectively). Additionally, CSF protein concentrations and TNCC in RDC- were lower than previously published reference ranges. Using an interpretation rule based on CSF protein concentration and TNCC, it was possible to accurately identify 13 RDC with spinal cord lesions and 6 RDC without lesions. It was not possible to determine spinal cord status in the remaining 18 RDC. CONCLUSIONS AND CLINICAL IMPORTANCE: Cerebrospinal fluid analysis is valuable in the evaluation of spinal cord status in RDC. The prognosis associated with these findings remains to be determined.

Cholangiohepatitis in Dairy Cattle: 13 Cases.

BACKGROUND: The signalment, clinicopathologic, bacteriological, histopathological, ultrasonographic characteristics, and the treatment and outcomes of adult cattle with cholangiohepatitis are poorly defined. ANIMALS: Thirteen Holstein cows with cholangiohepatitis. METHODS: Retrospective study of medical records of cattle admitted to the CHUV and the AVC between 1992 and 2012 and 2000 and 2012, respectively, for cattle older than 3 months of age with a histopathological diagnosis of cholangiohepatitis. Cholangiohepatitis was defined as the presence of portal inflammation surrounding or infiltrating bile ducts, with or without epithelial damage, and extending into the adjacent lobules. RESULTS: At the time of diagnosis of cholangiohepatitis, cows had decreased appetite (n = 7) or were anorectic (n = 6), had fever (n = 5), and had tachycardia (n = 8). Icterus was detected in 5 cows. Yellow discoloration was identified on the skin of the udder (n = 3), conjunctiva (n = 2), and vulva (n = 1). There was leukocytosis (n = 6), neutrophilia (n = 9), and hyperfibrinogenemia (n = 8). Alteration in the serum biochemistry profile included hyperglobulinemia (n = 8), hypoalbuminemia (n = 10), increased activity of GGT (n = 12), AST (n = 8), and ALP (n = 10), and hyperbilirubinemia (n = 10). Histopathological diagnosis included mild, subacute, nonsuppurative cholangiohepatitis (n = 4), mild suppurative cholangiohepatitis (n = 4), mild mixed (neutrophilic and lymphocytic) cholangiohepatitis (n = 3), and moderate, chronic, nonsuppurative cholangiohepatitis (n = 1). Six cows were discharged from the hospital, and 7 were euthanized. CONCLUSIONS AND CLINICAL IMPORTANCE: Cholangiohepatitis is a rare condition in adult cattle. Antemortem diagnosis can be challenging because clinical signs are unspecific.

RNAseq profiling of primary microglia and astrocyte cultures in near-term ovine fetus: A glial in vivo-in vitro multi-hit paradigm in large mammalian brain.

BACKGROUND: The chronically instrumented fetal sheep is a widely used animal model to study fetal brain development in health and disease, but no methods exist yet to interrogate dedicated brain cell populations to identify their molecular and genomic phenotype. For example, the molecular mechanisms whereby microglia or astrocytes contribute to inflammation in the brain remain incompletely understood. NEW METHOD: Here we present a protocol to derive primary pure microglial or astrocyte cultures from near-term fetal sheep brain, after the animals have been chronically instrumented and studied in vivo. Next, we present the implementation of whole transcriptome sequencing (RNAseq) pipeline to deeper elucidate the phenotype of such primary sheep brain glial cultures. RESULTS: We validate the new primary cultures method for cell purity and test the function of the glial cells on protein (IL-1ß) and transcriptome (RNAseq) levels in response to a lipopolysaccharide (LPS) challenge in vitro. COMPARISON WITH EXISTING METHODS: This method represents the first implementation of pure microglial or astrocytes cultures in fetal sheep brain. CONCLUSIONS: The presented approach opens new possibilities for testing not only supernatant protein levels in response to an in vitro challenge, but also to evaluate changes in the transcriptome of glial cells derived from a large mammalian brain bearing high resemblance to the human brain. Moreover, the presented approach lends itself to modeling the complex multi-hit paradigms of antenatal and perinatal cerebral insults in vivo and in vitro.

Diagnostic accuracy of refractometer and Brix refractometer to assess failure of passive transfer in calves: protocol for a systematic review and meta-analysis.

Calves are highly dependent of colostrum (and antibody) intake because they are born agammaglobulinemic. The transfer of passive immunity in calves can be assessed directly by dosing immunoglobulin G (IgG) or by refractometry or Brix refractometry. The latter are easier to perform routinely in the field. This paper presents a protocol for a systematic review meta-analysis to assess the diagnostic accuracy of refractometry or Brix refractometry versus dosage of IgG as a reference standard test. With this review protocol we aim to be able to report refractometer and Brix refractometer accuracy in terms of sensitivity and specificity as well as to quantify the impact of any study characteristic on test accuracy.

Increased blood-circulating interferon-γ, interleukin-17, and osteopontin levels in bovine paratuberculosis.

Paratuberculosis-infected cattle initially develop an effective cell-mediated immune response that declines as the disease progresses. Blood is one of best sources for characterizing the inflammatory status of infected cows and for studying mediators related to chronic diseases. The aim of this study was to evaluate the cow-level association between blood cytokine concentration, the influence of serum on immune cell proliferation, and dairy cows naturally infected with Mycobacterium avium ssp. paratuberculosis (MAP). Positive animals (n=41) from 19 herds were selected on the basis of 2 positive fecal culture results and divided into 2 groups: single-positive, or serum ELISA-negative cows (n=32), and double-positive, or cows that gave positive results for both mycobacterial culture and serum ELISA (n=9). Negative animals (n=39) were selected from paratuberculosis-negative herds in which at least 80% of the animals had been diagnosed as negative by fecal culture and ELISA and that did not produce positive results during the 2-yr study. Analysis of plasma levels of the cytokines IL-4, IL-10, IL-17, IFN-γ, and osteopontin was performed, revealing distinct patterns. The ELISA-positive cows with MAP shedding had similar plasma concentrations of IL-4 and IL-10 but elevated levels of IFN-γ, IL-17, and osteopontin, which is indicative of inflammatory disease in these subclinical positive cows. In vitro MAP infection of bovine macrophages showed increased gene expression of tumor necrosis factor-α, IL-1ß, IL-6, IL-23, and transforming growth factor-ß as early as 6h postinfection for all of the cytokines involved in the establishment of a T-helper type-17 immune response. To determine the systemic influence of serum on immune cell functions, lymphoproliferation assays were also performed in presence of JD serum. The serum from shedding cows showed 15% less proliferation. These results indicate that infected cows have a lower systemic capacity to maintain a protective immune response and that, as the disease progresses, an emerging T-helper type-17 immune response is established.

Neonatal morbidity and mortality of 31 calves derived from somatic cloning.

BACKGROUND: The neonatal period is associated with high morbidity and mortality in cloned calves. OBJECTIVE: To describe morbidity and mortality in cloned calves from birth to 2 years of age. ANIMALS: Thirty-one somatic cell-derived Holstein calves delivered at a veterinary teaching hospital. METHODS: Medical files were retrospectively analyzed. RESULTS: Four calves were stillborn. Five calves born alive had physical congenital defects. Twenty-three calves had an enlarged umbilical cord. Laboratory abnormalities included acidemia, respiratory acidosis, hyperlactatemia, anemia, stress leukogram, decreased total protein, albumin and globulins, and increased creatinine. Twenty-five calves survived the 1st hour of life. Among them, 11 stood without assistance within 6 hours of birth, 10 calves took longer than 6 hours to stand, and 4 never stood. Twenty-two calves suffered from anorexia. Twelve calves had complications arising from umbilical cord infections. Three calves developed idiopathic hyperthermia (>40°C). Eight calves suffered from gastrointestinal problems, including ruminal distension, abomasal ulcers, neonatal enteritis, intussusception, and abomasal displacement. Mortality between birth and 3 weeks of age was 32% (10/31). Causes of death and reasons for euthanasia included stillbirths, respiratory failure, and limb deformities. Mortality between 3 weeks and 2 years of age was 19% (4/21), with deaths in this group attributed to generalized peritonitis and complications arising from umbilical infections. Overall, mortality rate within 2 years of age was 14/31 (45%). CONCLUSION AND CLINICAL IMPORTANCE: Respiratory problems, limb deformities, and umbilical infections were the most common causes of morbidity and mortality in these cloned calves.

Developmental and epigenetic anomalies in cloned cattle.

Many of the developmental anomalies observed in cloned animals are related to foetal and placental overgrowth, a phenomenon known as the 'large offspring syndrome' (LOS) in ruminants. It has been hypothesized that the epigenetic control of imprinted genes, that is, genes that are expressed in a parental-specific manner, is at the root of LOS. Our recent research has focused on understanding epigenetic alterations to imprinted genes that are associated with assisted reproductive technologies (ART), such as early embryo in vitro culture (IVC) and somatic cell nuclear transfer (SCNT) in cattle. We have sought and identified single nucleotide polymorphisms in Bos indicus DNA useful for the analysis of parental-specific alleles and their respective transcripts in tissues from hybrid embryos derived by crossing Bos indicus and Bos taurus cattle. By analysing differentially methylated regions (DMRs) of imprinted genes SNRPN, H19 and the IGF2R in cattle, we demonstrated that there is a generalized hypomethylation of the imprinted allele and the biallelic expression of embryos produced by SCNT when compared to the methylation patterns observed in vivo (artificially inseminated). Together, these results indicate that imprinting marks are erased during the reprogramming of the somatic cell nucleus during early development, indicating that such epigenetic anomalies may play a key role in mortality and morbidity of cloned animals.

Risk factors associated with transmission of Mycobacterium avium subsp. paratuberculosis to calves within dairy herd: a systematic review.

BACKGROUND: Paratuberculosis has a worldwide distribution and many countries have implemented control programs to prevent transmission among and within herds. For these programs to be efficient, knowledge of the risk factors involved in transmission is essential. OBJECTIVES: Systematically review the scientific literature concerning risk factors associated with Mycobacterium avium subsp. paratuberculosis (MAP) transmission to dairy calves. STUDY DESIGN: Systematic review. METHODS: An electronic search was done in PubMed and CAB to retrieve references relevant to answer at least 1 of the 5 questions concerning neonatal environment, colostrum, milk, housing of calves, and contact of calves with adult cow feces as risk factors in MAP transmission. A 1st screening was done using titles only, then abstracts, and finally full-length articles were reviewed for relevance. From the articles selected, risk factors and presence of a significant association between these risk factors and MAP transmission were recorded. RESULTS: Twenty-three articles from 11 different countries and published in 12 different journals were reviewed. The most common study design was cross-sectional (n = 16). The case definition and diagnostic tests used were very variable among studies, but serum ELISA was used in most studies (n = 14). The study unit was the herd in 18 studies. CONCLUSIONS AND CLINICAL IMPORTANCE: The contact of calves with adult cow feces is the most important risk factor in MAP transmission. The 5 categories of risk factors are linked to one another.