The Vagus Nerve Regulates Immunometabolic Homeostasis in the Ovine Fetus near Term: The Impact on Terminal Ileum.

BACKGROUND: Glucosensing elements are widely distributed throughout the body and relay information about circulating glucose levels to the brain via the vagus nerve. However, while anatomical wiring has been established, little is known about the physiological role of the vagus nerve in glucosensing. The contribution of the vagus nerve to inflammation in the fetus is poorly understood. Increased glucose levels and inflammation act synergistically when causing organ injury, but their interplay remains incompletely understood. We hypothesized that vagotomy (Vx) will trigger a rise in systemic glucose levels and this will be enhanced during systemic and organ-specific inflammation. Efferent vagus nerve stimulation (VNS) should reverse this phenotype. METHODS: Near-term fetal sheep (n = 57) were surgically prepared using vascular catheters and ECG electrodes as the control and treatment groups (lipopolysaccharide (LPS), Vx + LPS, Vx + LPS + selective efferent VNS). The experiment was started 72 h postoperatively to allow for post-surgical recovery. Inflammation was induced with LPS bolus intravenously (LPS group, 400 ng/fetus/day for 2 days; n = 23). For the Vx + LPS group (n = 11), a bilateral cervical vagotomy was performed during surgery; of these n = 5 received double the LPS dose, LPS800. The Vx + LPS + efferent VNS group (n = 8) received cervical VNS probes bilaterally distal from Vx in eight animals. Efferent VNS was administered for 20 min on days 1 and 2 +/10 min around the LPS bolus. Fetal arterial blood samples were drawn on each postoperative day of recovery (-72 h, -48 h, and -24 h) as well as at the baseline and seven selected time points (3-54 h) to profile inflammation (ELISA IL-6, pg/mL), insulin (ELISA), blood gas, and metabolism (glucose). At 54 h post-LPS, a necropsy was performed, and the terminal ileum macrophages' CD11c (M1 phenotype) immunofluorescence was quantified to detect inflammation. The results are reported for p < 0.05 and for Spearman R2 > 0.1. The results are presented as the median (IQR). RESULTS: Across the treatment groups, blood gas and cardiovascular changes indicated mild septicemia. At 3 h in the LPS group, IL-6 peaked. That peak was decreased in the Vx + LPS400 group and doubled in the Vx + LPS800 group. The efferent VNS sped up the reduction in the inflammatory response profile over 54 h. The M1 macrophage activity was increased in the LPS and Vx + LPS800 groups only. The glucose and insulin concentrations in the Vx + LPS group were, respectively, 1.3-fold (throughout the experiment) and 2.3-fold higher vs. control (at 3 h). The efferent VNS normalized the glucose concentrations. CONCLUSIONS: The complete withdrawal of vagal innervation resulted in a 72-h delayed onset of a sustained increase in glucose for at least 54 h and intermittent hyperinsulinemia. Under the conditions of moderate fetal inflammation, this was related to higher levels of gut inflammation. The efferent VNS reduced the systemic inflammatory response as well as restored both the concentrations of glucose and the degree of terminal ileum inflammation, but not the insulin concentrations. Supporting our hypothesis, these findings revealed a novel regulatory, hormetic, role of the vagus nerve in the immunometabolic response to endotoxin in near-term fetuses.

Ability of unsterilized recycled manure solids bedding to support growth of Klebsiella pneumoniae and Escherichia coli.

Although recycled manure solids (RMS) bedding is used on dairy farms, it could allow bacterial growth when contaminated by feces and thus increase the incidence of clinical mastitis in cows. The objective of this study was to describe bacterial growth in three different types of RMS bedding, as well as in sand, when samples were experimentally inoculated with Escherichia coli and Klebsiella pneumoniae. Two 3-day trials were conducted, during which treatments included inoculating bedding samples with E. coli and K. pneumoniae, as well as no inoculation. The trial was repeated 3 times for each bedding sample on each day. Samples were incubated at 15°C for 3 d and bacterial counts were measured every day. After inoculation, there was no significant K. pneumoniae or E. coli growth phase during the trial in those RMS samples that were prepared either in a container or in a heap. Recycled manure solids and sand samples prepared in a rotary drum, however, showed a similar active growth phase of K. pneumoniae during the first 24 h of the trial. Moreover, a significant E. coli growth phase was observed in the samples of sand bedding in the first 24 h. The 3 different types of RMS bedding samples did not react in a similar manner to coliform inoculation. No active growth phase was observed in bedding samples already containing a high bacterial concentration following inoculation with coliforms.

Bien que la litière de fumier recyclé (LFR) soit utilisée dans les fermes laitières, elle pourrait permettre la croissance bactérienne lorsqu'elle est contaminée par des matières fécales et augmenter ainsi l'incidence de mammite clinique chez les vaches. L'objectif de cette étude était de décrire la croissance bactérienne dans trois types de LFR, ainsi que dans du sable, lorsque des échantillons étaient inoculés expérimentalement avec Escherichia coli et Klebsiella pneumoniae. Deux essais de trois jours ont été réalisés, au cours desquels les échantillons de litière ont été inoculés ou non avec E. coli et K. pneumoniae. L'essai a été répété trois fois pour chaque échantillon de litière, chaque jour. Les échantillons ont été incubés à 15 °C pendant 3 jours et la numération bactérienne a été mesurée chaque jour. Après inoculation, il n'y a pas eu de phase de croissance significative de K. pneumoniae ou d'E. coli au cours de l'essai dans les échantillons de LFR préparés dans un conteneur ou en tas. Les échantillons de sable et de LFR préparés dans un tambour rotatif ont cependant montré une phase de croissance active similaire de K. pneumoniae pendant les premières 24 heures de l'essai. En outre, une phase de croissance significative d'E. coli a été observée dans les échantillons de litière de sable au cours des premières 24 h. Les trois différents types d'échantillons de LFR n'ont pas réagi de la même manière à l'inoculation de coliformes. Aucune phase de croissance active n'a été observée dans les échantillons de litière contenant déjà une concentration bactérienne élevée après l'inoculation de coliformes.(Traduit par Docteur Simon Dufour).

Comparison of 2 PCR assays on environmental samples cultured for Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causal agent of paratuberculosis, a chronic, contagious, and incurable enteric disease of ruminants. An in-house IS900 PCR assay validated for MAP detection in sheep has been shown to have a higher sensitivity than a commercial PCR and fecal culture. We have now compared the performance of this in-house IS900 PCR assay with a commercial ISMap02 PCR assay for the detection of MAP DNA in bovine dairy farm environmental samples. We purposefully selected 30 culture-positive, 62 culture-negative, and 62 non-interpretable environmental samples. We applied the IS900 PCR assay directly to the frozen inoculum of these samples. Inocula were incubated in an automated system, and growth was confirmed by an acid-fast bacilli stain and the IS900 PCR assay. Among culture-positive samples before incubation, the IS900 PCR assay yielded significantly more positive results than the ISMap02 PCR assay; however, among culture-negative samples, the IS900 PCR assay yielded positive results both before and after incubation. The ISMap02 PCR assay did not flag positively among the culture-negative samples either before or after incubation. The IS900 PCR assay is a sensitive method that can be used to detect MAP DNA in environmental samples before incubation. The ISMap02 PCR assay is a specific method used to detect MAP DNA in environmental samples both before and after incubation.

Accuracy of testing strategies using antibody-ELISA tests on repeated bulk tank milk samples and/or sera of individual animals for predicting herd status for Salmonella dublin in dairy cattle.

There is currently no perfect test for determining herd-level status for Salmonella Dublin in dairy cattle herds. Our objectives were to evaluate the accuracy, predictive ability, and misclassification cost term of different testing scenarios using repeated measurements for establishing the S. Dublin herd status. Diagnostic strategies investigated used repeated bulk tank milk antibody-ELISA tests, repeated rounds of blood antibody-ELISA tests on non-lactating animals or a combination of both approaches. Two populations hypothesized to have different S. Dublin prevalences were included: (i) a convenience sample of 302 herds with unknown history of infection; and (ii) a cohort of 58 herds that previously tested positive to S. Dublin. Bulk milk samples were collected monthly for 6-7 months and serum were obtained from 10 young animals on two occasions, at the beginning and end of bulk milk sampling period. A series of Bayesian latent class models for two populations and comparing two tests were used to compare bulk milk-based to serum-based strategies. Moreover, Monte Carlo simulations were used to compared diagnostic strategies combining both types of samples. For each diagnostic strategy, we estimated the predictive values using two theoretical prevalences (0.05 and 0.25). Misclassification cost term was also estimated for each strategy using these two prevalences and a few relevant false-negative to false-positive cost ratios. When used for screening a population with an expected low prevalence of disease, for instance for screening herds with no clinical signs and no previous S. Dublin history, a diagnostic strategy consisting of two visits at 6 months interval, and with herd considered positive if bulk milk PP% ≥ 35 and/or ≥ 1/10 animals are positive on one or both visits could be used to confidently rule-out S. Dublin infection (median negative predictive value of 0.99; 95% Bayesian credible intervals, 95BCI: 0.98, 1.0). With this approach, however, positive results should later be confirmed with more specific tests to confirm whether S. Dublin is truly present (median positive predictive value of 0.36; 95BCI: 0.22, 0.57). The same diagnostic strategy could also be used confidently to reassess the S. Dublin status in herds with a previous S. Dublin history. When use for such a purpose, the predictive value of a positive result could be greatly improved, from 0.78 (95BCI: 0.65, 0.90) to 0.99 (95BCI: 0.94, 1.0) by requiring ≥ 1 positive result on both visits, rather than at any of the two visits.

Inter-Rater Reliability of Scoring Systems for Abomasal Lesions in Quebec Veal Calves.

The objective of this study was to determine the inter-rater reliability of current scoring systems used to detect abomasal lesions in veal calves. In addition, macroscopic lesions were compared with corresponding histological lesions. For this, 76 abomasa were retrieved from veal calves in a slaughterhouse in Quebec and scored by four independent raters using current scoring systems. The localisations of the lesions were separated into pyloric, fundic, or torus pyloricus areas. Lesions were classified into three different types, i.e., erosions, ulcers, and scars. To estimate the inter-rater reliability, the coefficient type 1 of Gwet's agreement and Fleiss κ were used for the presence or absence of a lesion, and the intra-class correlation coefficient was used for the number of lesions. All veal calves had at least one abomasal lesion detected. Most lesions were erosions, and most of them were located in the pyloric area. Overall, a poor to very good inter-rater agreement was seen for the pyloric area and the torus pyloricus regarding the presence or absence of a lesion (Fleiss κ: 0.00-0.34; Gwet's AC1: 0.12-0.83), although a higher agreement was observed when combining all lesions in the pyloric area (Fleiss κ: 0.09-0.12; Gwet's AC1: 0.43-0.93). For the fundic area, a poor to very good agreement was also observed (Fleiss κ: 0.17-0.70; Gwet's AC1: 0.90-0.97). Regarding the inter-rater agreement for the number of lesions, a poor to moderate agreement was found (ICC: 0.11-0.73). When using the scoring system developed in the European Welfare Quality Protocol, a poor single random rater agreement (ICC: 0.42; 95% CI: 0.31-0.56) but acceptable average random rater agreement (ICC: 0.75; 95% CI: 0.64-0.83) was determined. Microscopic scar lesions were often mistaken as ulcers macroscopically. These results show that the scoring of abomasal lesions is challenging and highlight the need for a reliable scoring system. A fast, simple, and reliable scoring system would allow for large scale studies which investigate possible risk factors and hopefully help to prevent these lesions, which can compromise veal calves' health and welfare.

Health indicators in surplus calves at the time of arrival at auction markets: Associations with distance from farms of origin in Québec, Canada.

This study evaluated the associations between estimated distance from farms' locations to auction markets, and health indicators of surplus dairy calves sold during summer 2019 and winter 2020 in Québec, Canada. A total of 3,610 animals from 1,331 different farms were used in this cross-sectional cohort study. Geographic coordinates (latitude and longitude) were obtained for each farm and the 2 participating livestock auction markets. Calves' abnormal physical signs (APS) were noted upon arrival at the auction market as they were examined by trained research staff. The haversine distance between the farm and the auction market was evaluated using geographic coordinates and categorized. Generalized linear mixed models were used for statistical analyses. The main APS observed were ocular discharge (34.9%), abnormal hide cleanliness (21.2%), swollen navel (17.2%), dehydration score 1 (at least one of the 2 following clinical signs: persistent skin tent or sunken eye, 12.9%), and dehydration score 2 (both clinical signs mentioned above, 6.5%). Calves from farms located at greater distances from the auction markets (≥110 km) had a higher risk ratio [RR = 1.08; 95% confidence internal (CI) = 1.03, 1.13] for dehydration than those from lesser distances (0-25 km). During the summertime, a RR of 1.18 (95% CI = 1.15, 1.22) was observed for dehydration compared with wintertime. A 2-way interaction between estimated distance and season showed a higher prevalence of ocular discharge for calves from farms at distances greater than or equal to 110 km during the summer (RR = 1.11; 95% CI: 1.04, 1.20) than for calves from farms located at lesser distances (0-25 km). These results demonstrate that calves from farms located at greater distances from the auction markets had more APS, mainly during the summer. A better understanding of the transport conditions and interaction with management at the farm of origin is determinant to mitigate the impact of the journey on surplus calf health.

Presence or severity of hypophosphatemia is not associated with survival outcome in postpartum downer dairy cows.

OBJECTIVE: To evaluate the association between serum phosphorus concentration and the outcome of postpartum downer cows. ANIMALS: Postpartum downer dairy cows presented over a 22-year period. PROCEDURES: In this cross-sectional study (1994 to 2016), medical records of all postpartum downer cows presented to a referral large animal hospital were reviewed. The association between serum inorganic phosphorus concentration and survival was assessed using a multivariable logistic regression. RESULTS: 907 postpartum downer dairy cows were included and classified as hypophosphatemic (mild: > 2.25 to < 3.25 mg/dL, moderate: > 1.50 to ≤ 2.25 mg/dL, and severe: ≤ 1.50 mg/dL), normophosphatemic (≥ 3.25 and ≤ 8.76 mg/dL) or hyperphosphatemic (> 8.76 mg/dL). Hypophosphatemia was observed in 19.4% of the cows (n = 176). Of those, 54.5% (n = 96) were also hypocalcemic. Overall, 58.4% cows (n = 530) survived after hospitalization. Hypophosphatemia was not significantly associated with the outcome of postpartum downer cows (mild: OR = 1.0, 95% CI: 0.6 to 1.8; moderate: OR = 0.5, 95% CI: 0.2 to 1.1; severe: OR = 1.0, 95% CI: 0.4 to 2.4). CLINICAL RELEVANCE: Low serum phosphorus concentration was frequently observed with hypocalcemia and was not associated with the outcome of postpartum downer cows.

Interpretation of cerebrospinal fluid analysis from recumbent cows using different thresholds of red blood cell count.

BACKGROUND: Hemodilution of the cerebrospinal fluid (CSF) could confound interpretation of results. Accurately predicting total nucleated cells count (TNCC) and total protein concentration (TPC) attributable to hemodilution is difficult. OBJECTIVE: To determine the effects of hemodilution on TPC and TNCC in bovine CSF. METHODS: Retrospective review of CSF analysis results of downer dairy cows treated at Centre hospitalier universitaire vétérinaire between January 2006 and December 2014. Descriptive statistics were performed using 3 scenarios. RESULTS: Among the 235 samples included, red blood cell (RBC) count (RBCC) ranged from 0 to 869 220 RBC/µL (median = 6.6), TPC ranged from 0.04 to 6.51 g/L (median = 0.27), and TNCC ranged from 0 to 7500 cell/µL (median = 1.1). Among the 157 samples that had <30 RBC/µL (a threshold used in other species), TPC and TNCC varied between 0.13 and 1.06 g/L (median = 0.27) and between 0 and 31.4 cell/µL (median = 0.6), respectively. Eighty-four samples had TPC <0.25 g/L and TNCC ≤4.5 cell/µL. Among those 84 samples, RBCC varied between 0 and 1290 RBC/µL (median = 4.7). In 20 samples, TNCC was 0 with a variation in RBCC between 0 and 840 RBC/µL (median = 3.9). No strong correlations between RBCC and TNCC and TPC were found. CONCLUSIONS: A cutoff around 200 RBC/µL is proposed as clinically meaninful in bovine CSF. Results between 200 and 1290 RBC/µL are equivocal.

Estimation of the accuracy of an ELISA test applied to bulk tank milk for predicting herd-level status for Salmonella Dublin in dairy herds using Bayesian Latent Class Models.

Enzyme-Linked Immunosorbent Assay (ELISA) test is commonly used for detection of antibodies to Salmonella Dublin in individual bovine milk samples. However, little is known about its accuracy when used on bulk tank milk for determining herd-level S. Dublin status and when evaluated without assuming a perfect reference test. The objectives of this study were: i) to estimate the herd prevalence of S. Dublin among dairy cattle herds in Québec, Canada; ii) to estimate the herd sensitivity and specificity of a commercially available ELISA test when used on bulk milk; iii) to examine how the diagnostic test accuracy varies with different bulk milk ELISA cut-offs; and (iv) to assess the added value of combining ELISA screening of bulk milk and individual serum of 10 animals for determining S. Dublin herd status. A cohort of 302 dairy herds selected in three regions (population 1) and 58 herds that have already tested positive to S. Dublin (population 2) were recruited. A total of 715 bulk milk samples and 7150 individual blood samples from cattle over 3 months old (10 animals per herd) sampled on two occasions were collected. Testing was conducted using PrioCHECK™ Salmonella Ab bovine Dublin ELISA test for milk (Bmilk ELISA: test under investigation) and for serum of 10 individual animals (Serum10 ELISA: imperfect reference test) to determine the herd-level S. Dublin status. A latent class model for two populations, two tests, allowing for conditional dependence between tests was fit within a Bayesian framework. At cut-off PP % ≥ 15 for a Bmilk ELISA, which is used by provincial authorities, the herd prevalence of S. Dublin estimated using informative prior was 6.8 % (4.3-9.9) in population 1. The herd sensitivity and specificity estimates (95 % Bayesian Credibility Intervals) for Bmilk ELISA were 40.6 % (15.6-88.8) and 91.9 % (88.3-95.8), respectively. Positive and negative predictive values of Bmilk ELISA applied in population 1 were 26.4 % (8.5-60.2) and 95.8 % (92.1-99.2), respectively. Increasing Bmilk ELISA cut-offs had little influence on predictive values. The combination of both ELISA tests did not improve the diagnostic accuracy of S. Dublin. Our study shows that a test-positive herd based on a single bulk milk sample would require complementary tests for status confirmation. However, a test-negative herd could be classified as true negative with a high certainty.

Corrigendum: Association Between Recycled Manure Solids Bedding and Subclinical Mastitis Incidence: A Canadian Cohort Study.

[This corrects the article DOI: 10.3389/fvets.2022.859858.].

Association Between Recycled Manure Solids Bedding and Subclinical Mastitis Incidence: A Canadian Cohort Study.

Recycled manure solids (RMS) are increasingly used as bedding for dairy cows. However, potential impact of RMS bedding on animal health is not well described. The objective of this study was to evaluate subclinical mastitis incidence in cows housed on RMS bedding. Twenty RMS farms and a comparative group of 60 straw-bedded farms were enrolled in a 1-year longitudinal study (2018-2019). Data from 11,031 dairy cows were collected. Variations of individual somatic cell count were evaluated using three different methods. First, we compared the cow's mean lactation linear score between cows housed on the two bedding types. Then, we compared across bedding types the risk for a given cow of having a milk test with a linear score ≥ 4. Finally, we evaluated the dynamics of somatic cell count using pairs of tests within a cow. More specifically, we considered that only pairs of DHI tests where the first test yielded a linear score < 4 were at risk of an incident subclinical mastitis event. Then, we defined a newly acquired subclinical mastitis when the second test was ≥ 4. All models were adjusted for putative confounders. We could not highlight a significant association between bedding type and cow's mean lactational linear score (least square mean of 2.47 in cows from RMS farms vs. 2.37 in straw farms; 95%CI for linear score's difference: -0.20, 0.40). Furthermore, we could not find an association between bedding type and the risk of a high linear score (≥ 4). For the latter, cows housed on RMS had 0.93 times the risk of having a high linear score than straw-bedded cows (%95 CI: 0.68, 1.28). Moreover, cows on recycled manure solids farms had 0.73 time the risk of acquiring subclinical mastitis when compared to straw-bedded farms. Again, this risk was not statistically significant (%95 CI: 0.54, 1.00). In our study, RMS bedding was not associated with subclinical mastitis, as measured by somatic cell count, when compared to cows housed on a more conventional bedding, straw bedding.

Impact of recycled manure solids bedding on hygiene and odds of hock lesions in dairy cows.

The use of recycled manure solids (RMS) as bedding for dairy cows has become more popular over the last decade. Once producers own the RMS producing equipment, they are not restricted in the amount of bedding they can use in stalls, due to its large availability and low production costs. Thus, they are usually able to provide a soft lying area for their cows. Nevertheless, the potential positive impact of RMS on cows' hygiene and odds of hock lesions is not clearly demonstrated. Objectives of this research were, therefore, to compare RMS-bedded cows' hygiene level and odds of hock lesions to straw-bedded cows (comparative group). To achieve this, an observational cross-sectional study was conducted in 27 RMS- and 61 straw-bedded herds. During the farm visit, 30 cows per herd were scored for hygiene of three body regions (udder, lower legs and flank/upper legs) using a score ranging from 1 to 4 (1 being the cleanest and 4 the dirtiest). In addition, both hocks were scored (0 to 3) to evaluate the presence of lesions (0 when there was no lesion and 3 when a swelling protrusion > 2.5 cm was present). Continuation-ratio models were used to compute the odds of having a given score to the odds of having a lower score. Recycled manure solids usage was associated with lower odds of having an udder hygiene score ≥3 [odds ratio (OR): 0.43; 95% CI: 0.20, 0.95] and of having a score of 4 (OR: 0.29; 95% CI: 0.09, 0.93). It was also associated, compared to straw, with cleaner lower legs. We observed lower odds of having a score ≥2 (OR: 0.45; 95% CI: 0.21, 0.98), ≥3 (OR: 0.16; 95% CI: 0.04, 0.66), or a score of 4 (OR: 0.07; 95% CI: 0.01, 0.40) in RMS-bedded herds. No statistically significant association could be highlighted between RMS usage and flank/upper legs hygiene. Furthermore, we did not observe any statistically significant associations between bedding type (RMS vs. straw) and odds of hock lesions. In conclusion, cows bedded with RMS had generally cleaner udders and cleaner lower legs than straw-bedded ones. No association was found between bedding type and odds of hock lesions.

Factors Determining the Success of the Chronically Instrumented Non-anesthetized Fetal Sheep Model of Human Development: A Retrospective Cohort Study.

Background The chronically instrumented non-anesthetized fetal sheep (CINAFS) model has been a mainstay of human fetal development research for the past 60 years. As a large "two for one" animal model, involving the instrumentation of the ewe and her fetus, the model poses challenges to implement de novo and maintain overtime at the highest standards of operating procedures to ensure ongoing performance. A common yet conventionally underreported issue researchers face is a high rate of animal loss. Here, we investigate what determines the success of the CINAFS model of human development. Methods We conducted a retrospective cohort analysis consisting of 82 experiments spanning the course of six years. Our team identified 10 variables that we anticipated were likely to influence the experimental outcome, such as the time of year, animal size, and surgical complexity. To evaluate the role of each variable in contributing to the success of the model, a binary logit regression analysis with a Fisher scoring optimization was fit to the data (SAS, V9 engine, release 3.8, SAS Institute, Cary, NC, USA). A higher predictive probability indicates a larger impact by the given variable on the outcome of the experiment. A Wald chi-squared analysis was run on the data to control for confounders and determine significance. Results The single variable identified in this study as determining the success of experiment outcomes using the CINAFS model is the experience level of the team. Conclusion The CINAFS model offers enormous potential to further our understanding of human fetal development and create interventional technologies related to fetal health. However, to improve experimental outcomes using the CINAFS model, stronger communication and training are needed. We discuss the implications of our findings for the successful implementation of this challenging yet scientifically advantageous animal model of human physiology.

Clinical Mastitis Incidence in Dairy Cows Housed on Recycled Manure Solids Bedding: A Canadian Cohort Study.

Bedding can affect mammary health of dairy cows. The objectives of this study were to evaluate clinical mastitis incidence in cows housed on recycled manure solids bedding and, more specifically, to determine which pathogens were involved. We followed 26 recycled manure solids farms and 60 straw-bedded farms as a comparative group during 1 year (2018-2019). For each episode of clinical mastitis, defined as a visual alteration of the milk, with or without local or systemic signs of infection, producers sampled aseptically the affected quarter, provided some details about the animal, and sent the sample to the research team at the Université de Montréal. We received and analyzed 1,144 milk samples. The samples were cultured according to the National Mastitis Council guidelines and the different colony phenotypes were subsequently identified with mass spectrometry. In 54.6% of CM cases, a single phenotype of bacteria was cultured (pure culture), while two different phenotypes were found in 16.8% of the samples (mixed culture), and no growth was observed in 14.4% of the samples. Samples with three or more phenotypes were considered contaminated and were not included in the pathogen-specific analyses (14.3% of the submitted samples). The most frequently identified bacterial species in pure and mixed culture in farms using recycled manure solids were Streptococcus uberis (16.0%), Escherichia coli (13.8%), Klebsiella pneumoniae (13.2%), Streptococcus dysgalactiae (6.2%), and Staphylococcus aureus (3.4%). In straw farms, the most frequent species were S. aureus (16.6%), S. uberis (11.0%), E. coli (9.1%), S. dysgalactiae (8.0%), and K. pneumoniae (1.1%). The incidence of clinical mastitis (all cases together) was not higher in recycled manure solids farms (14.0 cases/100 cow-year; 95% CI: 8.3-23.7) compared with straw-bedded farms (16.3 cases/100 cow-year; 95% CI: 9.0-29.6). However, K. pneumoniae clinical mastitis episodes were 7.0 (95% CI: 2.0-24.6) times more frequent in recycled manure solids farms than in straw farms. Adjusted least square means estimates were 1.6 K. pneumoniae clinical mastitis cases/100 cow-year (95% CI: 0.8-3.4) in recycled manure solids farms vs. 0.2 cases/100 cow-year (95% CI: 0.1-0.6) in straw-bedded farms. Klebsiella pneumoniae clinical mastitis is in general severe. Producers interested in this bedding alternative need to be aware of this risk.

Bayesian accuracy estimates of environmental sampling for determining herd paratuberculosis infection status and its association with the within-herd individual fecal culture prevalence in Québec dairies.

The objectives of this retrospective analysis were to: 1) estimate the diagnostic sensitivity (Se) and specificity (Sp) of bacterial culture of environmental samples for determining Mycobacterium avium subsp. paratuberculosis (MAP) infection status in Québec dairies, using a Bayesian Latent Class Model (BLCM); and 2) determine the association between the number of positive environmental samples and the individual fecal culture (IFC) apparent and true MAP within-herd prevalence. Environmental and individual fecal samples were collected from 87 commercial dairy herds participating in previous research projects. Environmental samples included two composite samples of 20 g collected from different locations within each of the following sites: an area where manure from the majority of adult cattle accumulates, a manure storage area and another site of manure accumulation chosen by the veterinarian. Samples were cultured using the MGIT Para TB culture liquid media and the BACTEC MGIT 960 system. The Se and Sp of environmental sampling were estimated using a one-test-one-population BLCM. Herds were considered positive for environmental sampling if at least one out of the six samples collected was positive. The apparent and true IFC within-herd MAP prevalence estimates for each herd were obtained using a two-stage cluster BLCM, then merged in a single dataset with the environmental sample results. The association between the within-herd MAP prevalence results (apparent and true), and the number of positive environmental samples was assessed using a zero-inflated negative binomial (ZINB) model. In all BLCMs, median posterior estimates and 95 % Bayesian credible intervals (BCI) were obtained with OpenBUGS statistical freeware. Se and Sp of environmental sampling were 43.7 % (95 % BCI: 32.5-55.5) and 96.2 % (95 % BCI: 84.2-99.8), respectively. Overall, the number of positive environmental samples increased with the apparent and true MAP within-herd prevalence. The true prevalence was higher than the apparent prevalence for a given number of positive environmental samples. The probability of not observing a positive environmental sample decreased with the prevalence. Despite its imperfect accuracy, environmental sampling is an inexpensive and non-invasive sampling method to determine MAP infection status in tie-stall herds that can be used as a proxy to estimate the true within-herd prevalence. The absence of positive environmental samples in a single sampling visit is likely an indicator of a very low within-herd prevalence rather than being MAP exempt.

Survival and prognostic indicators in downer dairy cows presented to a referring hospital: A retrospective study (1318 cases).

BACKGROUND: Downer cow syndrome, a common problem in dairy cattle, represents a diagnostic and therapeutic challenge for the attending veterinarian. Identifying prognostic indicators and assessing the odds of survival may improve the accuracy of the clinician's prognosis at the time of diagnosis. OBJECTIVE: To describe a population of downer dairy cows referred to a hospital and investigate predictors of outcome. ANIMALS: Recumbent adult dairy cows (cows unable or unwilling to stand without help) treated at a referral hospital. METHODS: Data at the time of admission were collected from medical records of downer dairy cows treated at the Centre Hospitalier Universitaire Vétérinaire between 1994 and 2016. Simple and multivariable logistic regression analyses were performed to assess the association of predictors with the outcome. RESULTS: Among 1318 cows included, 727 (55%) cows were discharged, and 591 (45%) cows died or were euthanized. Cows with longer time of recumbency before referral (odds ratio [OR] = 3.6), tachycardia (100-120 beats per minute [bpm], OR = 1.93; >120 bpm, OR = 2.92), tachypnea (OR = 1.76), hypothermia (OR = 2.08), anemia (OR = 3.30), neutropenia (OR = 1.7), high aspartate aminotransferase activity (500-1000 U/L, OR = 2.16; >1000 U/L, OR = 6.69), and increased serum creatinine concentration (OR = 1.75) had higher odds of nonsurvival. CONCLUSIONS AND CLINICAL IMPORTANCE: These findings may help the practitioner to consider treatment options and decide if referral is likely beneficial based on the odds of success. Early recognition of low chance of survival may facilitate an early decision for euthanasia.

Recording and manipulation of vagus nerve electrical activity in chronically instrumented unanesthetized near term fetal sheep.

BACKGROUND: The chronically instrumented pregnant sheep has been used as a model of human fetal development and responses to pathophysiologic stimuli. This is due to the unique amenability of the unanesthetized fetal sheep to the surgical placement and maintenance of catheters and electrodes, allowing repetitive blood sampling, substance injection, recording of bioelectrical activity, application of electric stimulation, and in vivo organ imaging. Recently, there has been growing interest in the pleiotropic effects of vagus nerve stimulation (VNS) on various organ systems such as innate immunity and inflammation, and metabolism. There is no approach to study this in utero and corresponding physiological understanding is scarce. NEW METHOD: Based on our previous presentation of a stable chronically instrumented unanesthetized fetal sheep model, here we describe the surgical instrumentation procedure allowing successful implantation of a cervical uni- or bilateral VNS probe with or without vagotomy. RESULTS: In a cohort of 68 animals, we present the changes in blood gas, metabolic, and inflammatory markers during the postoperative period. We detail the design of a VNS probe which also allows recording from the fetal nerve. We also present an example of fetal vagus electroneurogram (VENG) recorded from the VNS probe and an analytical approach to the data. COMPARISON WITH EXISTING METHODS: This method represents the first implementation of fetal VENG/VNS in a large pregnant mammalian organism. CONCLUSIONS: This study describes a new surgical procedure allowing to record and manipulate chronically fetal vagus nerve activity in an animal model of human pregnancy.

Comparison of thoracic ultrasonography and thoracic radiography to detect active infectious bronchopneumonia in hospitalized dairy calves.

BACKGROUND: The best test between thoracic ultrasonography (TUS) and thoracic radiography (TR) or the best combination of tests (series or parallel) to detect active infectious bronchopneumonia (BP) in hospitalized dairy calves remains unknown. HYPOTHESIS/OBJECTIVES: To estimate performances of TUS and TR to detect active BP in hospitalized dairy calves and to determine the best strategy for using these tests based on a panel diagnosis method (PDM). Performances of TUS and TR were hypothesized to be equivalent. ANIMALS: Fifty hospitalized dairy calves (≥7 days old; ≤100 kg; standing; pCO2 ≥ 53 mm Hg; any reason of presentation). METHODS: Each calf prospectively and sequentially underwent physical examination, thoracic auscultation, blood analyses, and TUS and TR. Three blinded experts determined whether active BP was present/absent based on PDM. Krippendorff's alpha measured interexpert agreement. The sensitivities (Se) and specificities (Sp) of TUS and TR alone and in series or parallel were compared (McNemar's test; P < .05). RESULTS: Interexpert agreement was moderate at 0.58 (95%CI: 0.42; 0.73). The Se and Sp of TUS were 0.84 (95%CI: 0.60; 0.97) and 0.74 (95%CI: 0.57; 0.86), respectively. The Se and Sp of TR were 0.89 (95%CI: 0.67; 0.99) and 0.58 (95%CI: 0.39; 0.75), respectively. No significant difference was found in the Se and Sp of TUS and TR when analyzed alone, in series or in parallel. CONCLUSION: Thoracic ultrasonography or TR alone equally detected active BP in hospitalized dairy calves. Series or parallel analysis provided no additional benefit. Its ease of use and widespread accessibility support using TUS as a first-line test to detect active BP in hospitalized dairy calves.

Scoping review on clinical definition of bovine respiratory disease complex and related clinical signs in dairy cows.

Bovine respiratory disease complex (BRD) is a worldwide multifactorial infectious disease. Antimicrobials are commonly used for treating BRD because bacteria are often involved. The clinical diagnosis of BRD is a challenge, especially in adult dairy cows, where information on this syndrome is scant. Having a definition based on consistent and reliable clinical signs would improve the accuracy of BRD diagnosis and could help to develop an optimal treatment approach by an early detection. The aim of this scoping review was to review clinical signs that could be recognized by producers in dairy cattle suffering from naturally occurring infectious respiratory disease, as reported in the literature. A review of the literature was performed for articles published between January 1, 1990 and January 1, 2020. The search of literature in English, French, and Italian languages included 2 different databases (Pubmed, https://pubmed.ncbi.nlm.nih.gov/; CAB abstract, https://www.cabi.org/publishing-products/cab-abstracts/). Clinical signs were categorized as follows: (1) "general manifestations of disease," which included behavioral changes or fever; (2) "alterations in respiratory function," which included clinical signs specifically associated with the respiratory tract examination; and (3) "clinical signs of other body systems," which included clinical signs related to other systems such as diarrhea or subcutaneous emphysema. The focus of the review was on clinical signs that could be monitored by animal handlers and producers. A total of 1,067 titles were screened, and 23 studies were finally included. The most common general clinical signs were increased body temperature (reported in 83% of studies, n = 19), change in feed intake (26%, n = 6), altered mentation (22%, n = 5), and decreased milk production (17%, n = 4). The alterations in respiratory function noted were nasal discharge (74%, n = 17), cough (65%, n = 15), altered respiratory dynamic or dyspnea (61%, n = 14), increased respiratory rate (43%, n = 10), and ocular discharge or lacrimation (30%, n = 7). The clinical signs associated with infectious respiratory disease reported in the 23 studies generally lacked a clear description of what constitutes a deviation from normality (0-50% of studies clearly reported what was considered normal versus abnormal depending on the clinical signs). This limitation prevented any comparison between studies that apparently reported the same "clinical sign," but possibly referred to a different assessment and definition of what was considered normal versus abnormal. Therefore, the definition of clinical signs in a repeatable way with validated interobserver agreement to determine the optimal combination for the diagnosis of BRD in dairy cows is needed. This could lead to a more judicious use of antimicrobials for respiratory disease in adult dairy cows.