Methods used by Canadian dairy farmers for on-farm euthanasia and the emotions associated with the decision and the practice of euthanasia.

On-farm euthanasia is complicated in that farmers have to make the decision to end the life of an animal they are caring for. Studies have shown that some culling decisions are made too late, which results in animal welfare issues. However, information about on-farm euthanasia methods and the decision process leading to the euthanasia of an animal is limited. Additionally, emotions and feelings are involved in the decision and the act of euthanasia. The objectives of this study were to describe current practices related to on-farm euthanasia performed by Canadian dairy farmers and to assess the emotions and complex feelings surrounding the act. A cross-sectional study design was used to gather this information through an online questionnaire completed exclusively by Canadian dairy farmers. Data were analyzed descriptively using frequencies, and associations between the participants' characteristics and their practices and emotions regarding on-farm euthanasia were assessed using logistic regression models. A total of 479 dairy farmers from 5 provinces answered the survey. The most commonly used primary method of on-farm euthanasia was a firearm (>70%). Unacceptable euthanasia methods, such as standard 0.22-caliber long rifle for adult cows or no adjunct method following the use of captive bolt, were used by 25%, 18%, and 58% of the participants for calves, replacement animals, and dairy cows, respectively. Ninety-four percent of the participants reported that the farm owner was the person who always or often made the euthanasia decisions. In addition, 32% of the participants reported that the veterinarian always examined the animal before performing euthanasia, 51% reported at least one person received training to perform euthanasia, and 16% reported having a decision tree for euthanasia. Some participants (17%) were troubled with the responsibility for ending the life of their animals, and 19% perceived other people on the farm to be uncomfortable with performing euthanasia. Half of the participants reported that people performing euthanasia on the farm felt at least one related painful emotion or complex feeling, and half reported they had at least one reason to feel anxiety or unease concerning euthanasia. The results of this study showed gaps such as the limited use of acceptable euthanasia methods and decision trees by Canadian dairy farmers, low availability of on-farm training for euthanasia, and variable involvement of veterinarians.

Graduate Student Literature Review: What is known about the eliminative behaviors of dairy cattle?

The eliminative behaviors of dairy cattle include frequencies and distribution over time and space for defecations and urinations, how the animal responds to cow-related and environmental factors by way of altered patterns of defecation and urination, and how an animal carries out and responds to its own acts of elimination. This review discusses the available literature to first define and describe eliminative behaviors of dairy cattle; what follows is a discussion on what can affect eliminative behaviors and methods for managing them. Information regarding these behaviors is sparse for dairy cattle and is largely centered around frequencies and distributions over the day. Relationships exist between eliminative behaviors and activity levels of the animals and activity levels of the people who manage them, suggesting that types of housing systems play a key role in mainly where and when eliminations occur. It also seems that individual animals vary in their elimination frequencies, in which case it may be interesting to determine what aspects of their individuality contribute to these differences. Although aspects of housing are intended to separate animals from their excreta, stalls or cubicles are not necessarily designed with cattle's natural eliminative behaviors in mind. Refining the timing of management routines and training of animals are some options in the next steps toward managing eliminative behaviors.

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).

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.

Short communication: Efficacy of a second intrauterine cephapirin infusion for the treatment of purulent vaginal discharge and endometritis in postpartum dairy cows.

The objective of this study was to quantify the efficacy of a second intrauterine cephapirin treatment administered 14 d after the initial one on subsequent reproductive performance of postpartum dairy cows affected by purulent vaginal discharge (PVD) or endometritis (ENDO). In total, 4,140 Holstein cows from 30 commercial herds were enrolled in a randomized clinical trial. At 36 (±7) d in milk, cows were examined using the Metricheck device to diagnose PVD. An endometrial cytology sample was also collected from each cow to perform a leukocyte esterase test for diagnosing ENDO. Diagnosis of PVD and ENDO was done cow-side. Cows diagnosed with PVD or ENDO were assigned to receive 1 of 2 treatments: (1) a single intrauterine cephapirin infusion (500 mg of cephapirin benzathin; Metricure, Merck Animal Health, Kirkland, QC, Canada) at the time of initial examination or (2) a single intrauterine infusion at the time of initial examination and a second one 14 d later. Subsequent reproductive and culling events were collected until 200 d in milk. Statistical analyses were performed using univariable and multivariable mixed logistic regression models. In cows affected by PVD, a second intrauterine cephapirin infusion increased the pregnancy risk at first insemination in comparison with cows that only received one treatment (28.0 vs. 38.8%). In cows affected by ENDO, a second treatment also increased the pregnancy risk at first insemination compared with cows that only received one treatment (30.3 vs. 39.2%). Overall, these results demonstrate that administering a second intrauterine cephapirin infusion 14 d after the initial treatment in postpartum cows affected by PVD or ENDO did improve their subsequent reproductive performance.

Randomized controlled trial of pegbovigrastim as an adjunct therapy for naturally occurring severe clinical mastitis cases in dairy cows.

The main objective of this study was to quantify the effect of pegbovigrastim (PEG) as an adjunct therapy for naturally occurring severe mastitis cases on survival, intramammary bacteriological cure, and subsequent milk production. A double-blinded, randomized controlled trial was conducted on a single commercial dairy farm. During the study period, all cows having a case of severe clinical mastitis, defined as the presence of abnormal milk and inflammation in one quarter or more combined with the presence of systemic signs (pyrexia, dehydration, or recumbency), were enrolled in the study. In addition to a standardized therapy combining systemic and intramammary antimicrobials as well as systemic anti-inflammatory drugs, cows received a subcutaneous injection of either 2.7 mL of 0.9% sterile saline (control group; CON) or 2.7 mL of PEG (PEG group). A milk sample for bacteriological analysis was taken before treatment was administered and a second sample was taken 14 d after enrollment. Survival (30 d post-treatment) and bacteriological cure (14 d post-treatment) were analyzed by survival analysis and chi-squared tests, respectively, whereas daily milk production was treated as a repeated measure in mixed regression models. The 77 cows enrolled in the study were of parity 2 to 4 (mean = 3) and between 3 and 302 d in milk (mean = 154). The bacteria identified in milk at enrollment were Klebsiella spp. (n = 48; 62%), Escherichia coli (n = 16; 21%), Enterobacter spp. (n = 10; 13%), or no growth (n = 3; 4%). The probability of survival during the first 30 d after treatment was higher in the PEG group (84%) than in the control group (46%). Daily milk production over the 30-d period following treatment and bacteriological cure 14 d post-treatment did not differ between groups. Overall, cows treated with PEG as an adjunct therapy for naturally occurring cases of severe clinical mastitis had a better survival than cows treated with saline.

Short communication: Accuracy of estimation of lameness, injury, and cleanliness prevalence by dairy farmers and veterinarians.

Lameness, injuries, and cleanliness are considered important indicators of dairy cow welfare, milk production, and milk quality. Previous research has identified that farmers globally underestimate the prevalence of these cow-based measurements, but no information on the perceptions of veterinarians is available. Because veterinarians are often perceived as the main providers of health advice on farms, the objective of the present study was to evaluate the relationship between the true prevalence of lameness, injury (hock, knee, neck), and cleanliness (udder, legs, flanks), and the estimated prevalence of these issues by farmers and their herd veterinarians. A cross-sectional study was conducted between February 2016 and July 2017. First, the farm owner and the herd veterinarian were asked to estimate the prevalence of lameness, of neck, knee and hock injuries, and of udder, leg, and flank cleanliness on the farm. The research team then visited the farm and scored all lactating cows in the herd for each measurement. Linear regression models were used to assess the relationship between the prevalence estimated by the veterinarians and the farmers, of each cow-based measurement, and the true prevalence on the farm. The 93 herds enrolled had a median of 55 milking cows and were housed in tiestall (90%) and freestall (10%) barns. Ten herd veterinarians participated and were involved with 2 to 22 enrolled farms each. A wide variation was detected in the true prevalence of the different cow-based measurements among herds (lameness: range = 19-72%, median = 36%; neck injuries: range = 0-65%, median = 14%; knee injuries: range = 0-44%, median = 12%; hock injuries: range = 0-57%, median = 25%; dirty udder: range = 0-55%, median 13%; dirty legs: range = 0-91%, median = 18%; and dirty flanks: range = 0-82%, median = 20%). For both veterinarians and farmers, the perception of each cow-based measurement prevalence increased incrementally as the herd's true prevalence increased. Overall, farmers and veterinarians underestimated cow-based measurements. Farmers and veterinarians more accurately estimated lameness prevalence in herds with higher prevalence than in herds with low prevalence, suggesting a better awareness of the issue on farms with lameness problems. Injuries were less accurately estimated in herds with higher injury prevalence compared with herds with lower prevalence, suggesting an opportunity for better knowledge transfer in this area.

Associations between on-farm cow welfare indicators and productivity and profitability on Canadian dairies: II. On tiestall farms.

The use of tiestall housing for dairy cows is often criticized due to the reduced freedom of movement it offers for the animals. Maximizing comfort is especially important in tiestall farms to ensure an acceptable level of cow welfare. Motivating dairy producers to make financial investments directly aimed toward the improvement of their animals' welfare can be challenging, especially when financial returns are uncertain. The aim of this study was to evaluate the existence of associations between on-farm animal welfare and indicators of farm productivity and profitability in tiestall farms. The prevalence of animal-, resource-, and management-based welfare indicators was collected on 100 Canadian tiestall farms during a cow comfort study. Records from the dairy herd improvement agency were retrieved and used to calculate the farms' productivity and profitability measures. Univariable and multivariable linear regressions were used to assess the associations between welfare indicators and milk production, milk quality, cow longevity, and economic margins calculated over replacement costs. Increased yearly average corrected milk production was associated with longer average lying time [ß = 272; 95% confidence interval (CI): 94, 450] and a higher proportion of cows fitting the tie-rail height (ß = 6; 95% CI: 1, 11). Lower yearly average somatic cell count was associated with lower percentages of stalls mostly soiled with manure (ß = -3.7; 95% CI: -1.9, -5.4) and a lower proportion of cows with body condition score ≤2 (ß = -5.1; 95% CI: -2.3, -8.3). The average margin per cow over replacement costs was positively associated with average lying time (ß = 147; 95% CI: 27, 267), percent of stall not soiled with manure (ß = 7.2; 95% CI: 3.0, 11.3), and the frequency of scheduled hoof trimming per year. Some of the relationships found included interactions between animal- and management-based welfare measures. For example, the relationship between lameness prevalence and average milk production was modified through the milk production genetic index. Overall, the results show that improved cow comfort and welfare on tiestall farms is associated with increased productivity, cow longevity, and profitability when estimated through margins calculated over the replacement costs. Producers should aim to optimize all aspects of stall comfort to enhance their cows' productivity.

Associations between on-farm animal welfare indicators and productivity and profitability on Canadian dairies: I. On freestall farms.

Motivating dairy producers to financially invest in the improvement of their animals' comfort and welfare can pose some challenges, especially when financial returns are uncertain. Economic advantages for dairy producers associated with increased animal welfare are likely to come from either a premium paid for the milk or increased productivity. The aim of the current study was to evaluate the associations between measures of herd productivity and farm profitability and animal-, management-, and resource-based indicators of cow welfare and comfort. The cow welfare measures were collected during a cow comfort assessment conducted on 130 Canadian freestall dairy farms, including 20 using an automatic milking system. Herd productivity and farm profitability measures were retrieved or calculated from data collected by the regional dairy herd improvement programs, and included milk production and quality, longevity, and economic margins over replacement costs. Univariable and multivariable linear regression models were used to assess the associations between welfare indicators and productivity and profitability measures. Increased yearly corrected milk production was associated with reduced prevalence of cows with knee lesions [ß = 7.40; 95% confidence interval (CI): 2.6, 12.2], dirty flanks (ß = 26.9; 95% CI: 7.4, 46.5), and lameness (ß = 11.7; 95% CI: 3.3, 20.1). The farms' economic margin per cow, calculated over replacement costs, was associated with the within farm average lying time standard deviation (ß = -7.2; 95% CI: -12.7, -1.7), percent of stalls with dry bedding (ß = 6.4; 95% CI: 1.4, 11.4), and prevalence of cows with knee lesions (ß = -5.1; 95% CI: -8.9, -1.3). Some of the relationships found were complex, including several interactions between the animal-, management-, and resource-based measures. Overall, the results suggest that improved cow comfort and welfare on freestall farms is associated with increased herd productivity and profitability, when the latest is calculated by the margins over the replacement costs.

Is the profitability of Canadian tiestall farms associated with their performance on an animal welfare assessment?

In order for dairy producers to comply with animal welfare recommendations, financial investments may be required. In Canada, a new dairy animal care assessment program is currently being implemented under the proAction Initiative to determine the extent to which certain aspects of the Code of Practice are being followed and to assess the care and well-being of dairy cattle on farm. The aim of the current study was to evaluate the association between meeting the proAction animal-based and the electric trainer placement criteria and certain aspects of productivity and profitability on tiestall dairy farms. The results of a previous on-farm cow comfort assessment conducted on 100 Canadian tiestall farms were used to simulate the results of a part of the proAction Animal Care assessment on these farms. Each farm's productivity and profitability data were retrieved from the regional dairy herd improvement associations. Univariable and multivariable linear regressions were used to evaluate the associations between meeting these proAction criteria and the farms' average yearly: corrected milk production, somatic cell count (SCC), calving interval, number of breedings/cow, culling rate, prevalence of cows in third or higher lactation, and margins per cow and per kilogram of quota calculated over replacement costs. The association between milk production and the proAction lameness criterion was moderated through an interaction with the milk production genetic index which resulted in an increase in milk production per year with increasing genetic index that was steeper in farms that met the proAction lameness criterion compared with farms that did not. Meeting the proAction body condition score criterion was associated with reduced SCC and meeting the proAction electric trainer placement criterion was associated with SCC through an interaction with the farms' average SCC genetic index. The increase in SCC with increasing SCC genetic index was milder in farms that met this criterion compared with farms that did not. Farms that met the proAction electric trainer placement criterion had 4.6% more cows in their third or greater lactation. These results suggest that some associations exist between the productivity of Canadian tiestall farms and meeting several parameters of the proAction Animal Care assessment. Meeting these criteria is unlikely to impose any economic burden to the dairy industry as a whole.

Is the profitability of Canadian freestall farms associated with their performance on an animal welfare assessment?

Improving animal welfare on farm can sometimes require substantial financial investments. The Canadian dairy industry recently updated their Code of Practice for the care of dairy animals and created a mandatory on-farm animal care assessment (proAction Animal Care). Motivating dairy farmers to follow the recommendations of the Code of Practice and successfully meet the targets of the on-farm assessment can be enhanced by financial gain associated with improved animal welfare. The aim of the current study was to evaluate the association between meeting or not meeting several criteria from an on-farm animal welfare assessment and the farms' productivity and profitability indicators. Data from 130 freestall farms (20 using automatic milking systems) were used to calculate the results of the animal care assessment. Productivity and profitability indicators, including milk production, somatic cell count, reproduction, and longevity, were retrieved from the regional dairy herd improvement association databases. Economic margins over replacement costs were also calculated. Univariable and multivariable linear regression models were used to evaluate the associations between welfare and productivity and profitability indicators. The proportion of automatic milking system farms that met the proAction criterion for hock lesions was higher compared with parlor farms and lower for the neck lesion criterion. The proAction criterion for lameness prevalence was significantly associated with average corrected milk production per year. Average days in milk (DIM) at first breeding acted as an effect modifier for this association, resulting in a steeper increase of milk production in farms that met the criterion with increasing average DIM at first breeding. The reproduction and longevity indicators studied were not significantly associated with meeting or not meeting the proAction criteria investigated in this study. Meeting the proAction lameness prevalence parameter was associated with an increased profitability margin per cow over replacement cost by $236 compared with farms that did not. These results suggest that associations are present between meeting the lameness prevalence benchmark of the Animal Care proAction Initiative and freestall farms' productivity and profitability. Overall, meeting the animal-based criteria evaluated in this study was not detrimental to freestall farms' productivity and profitability.

Systematic early obstetrical assistance at calving: II. Effects on dairy heifer calf growth, health, and survival to weaning.

Good calving management should not only ease the transition of cows into lactation, but also contribute to providing healthy replacement animals for the herd. Difficulty during parturition has been found to be detrimental to the offspring. Because of the association of obstetrical intervention with undesirable outcomes, the general management recommendation for calving is to let cows give birth unassisted whenever possible. Unfortunately, very few studies have investigated the effects of planned early assistance during calving, regardless of whether or not another problem exists. To investigate the effects of early assistance during apparently normal calvings, a clinical trial was conducted on 257 Holstein cows. They were observed through the second stage of calving, and before calving they were divided randomly between 2 assigned interventions: not assisted or early assistance during the second stage of parturition. After calving, the animals were classified into 4 actual calving intervention groups: too quick to be assisted, not assisted, early assistance, and late assistance (for cows in the not assisted group that did not calve unassisted within the 1 h maximum time allowed). Early assistance was given 15 min after first sight of both front hooves of the calf and done using human force only. Heifer calves (n=129) born from enrolled dams were followed until weaning to assess the effect of assigned and actual calving interventions on their growth, health, and survival. Heifer calves' weight at birth was positively associated with weight at 7 wk, but the effect varied by assigned and actual intervention. As a routine management practice, giving early assistance during calving to dairy cows did not influence average daily gain, health, or survival in dairy heifer calves up to weaning at 7 wk of age.

Systematic early obstetrical assistance at calving: I. Effects on dairy calf stillbirth, vigor, and passive immunity transfer.

A critical time for dairy cattle is the perinatal period. Good calving management is critical to reduce periparturient losses and ensure the health of the offspring. Generally, it has been recommended that cows be allowed to calve unassisted when possible, but very few studies have been published that support or refute this general guideline. To investigate the effect of early assistance, a clinical trial enrolled 257 Holstein cows that were observed through the second stage of calving and assigned randomly to 1 of 2 calving interventions: not assisted (NA) or early assistance (EA) during the second stage of parturition. Early assistance was given 15min after the first sight of both front hooves of the calf and done using only human force. After calving, the animals were classified into 4 actual calving intervention groups: too quick to be assisted (TQ), NA, EA, and late assistance (LA; for cows in the NA group that did not calve unassisted within the 1h maximum time frame allowed). Giving early assistance to cows during calving as a routine management practice (assigned intervention) did not negatively influence calves' stillbirth risk, vigor at birth, or transfer of passive immunity. Calves in the LA intervention group had significantly greater odds of stillbirth than calves in the NA and EA groups, respectively. Calves in the LA group also had significantly worse vigor at birth than calves in the TQ, NA, or EA groups. Early assistance given at calving to cows that did not present signs of calving difficulties did not adversely affect calves' likelihood of being stillborn, vigor at birth, or transfer of passive immunity.

Calving management practices on Canadian dairy farms: Prevalence of practices.

Little information is available about current practices around calving in dairy cattle. The aim of this study was to describe calving management practices in the Canadian dairy industry related to housing, calving protocols, monitoring of parturition, and calving assistance. Information was gathered by in-person interviews from 236 dairy farms from 3 Canadian provinces (Alberta, Ontario, and Québec) with freestalls and an automatic milking system (n=24), freestalls with a parlor (n=112), and tiestalls (n=100). The most commonly used types of calving facilities were group calving pens (35%) followed by individual calving pens (30%). Tiestalls were used by 26% of all surveyed producers as their main type of calving area (49% of the tiestall, 7% of the freestall with parlor, and 13% of the automatic milking system farms). Written protocols related to calving were found on only 7% of the farms visited, and only 50% of those protocols were developed with a veterinarian. However, 90% of producers kept written records of calving difficulty. Monitoring of cows around calving occurred 5 times more often during the daytime (between morning and evening milking) compared with nighttime. Cameras were used to monitor cows around and during calvings on 18% of farms. Sixteen percent of producers vaginally palpated all animals during calving. Twenty-seven percent of producers interviewed assisted all calvings on their farms by pulling the calf, and 37% assisted all heifers at calving. According to the producers' reported perception, 93% of them had "a minor problem" or "no problem" with calving difficulties on their farms. This study provides basic data on current calving practices and identifies areas for improvement and potential targets for knowledge transfer efforts or research to clarify best management practices.

Addition of gut active carbohydrates to colostrum replacer does not improve passive transfer of immunoglobulin G in Holstein dairy calves.

The primary objective of this study was to investigate the effects of supplementing a commercial colostrum replacer (CR) with gut active carbohydrates (GAC) on passive transfer of IgG in commercial dairy calves. A secondary objective was to evaluate the effect of treatment on preweaning health and growth. A total of 240 newborn Holstein dairy calves on a commercial dairy farm were enrolled in this study. Newborn heifer and bull calves were weighed and then randomly assigned to either the treated group [GAC: 30g of GAC mixed into 1.5 doses (150g of IgG) of commercial colostrum replacer; n=119] or the control group [CON: 1.5 doses (150g of IgG) of CR; n=121]. The assigned CR treatment was fed within 3.5h of birth using an esophageal tube feeder. Venous blood samples were collected at 0 and 24h of age and used to measure serum IgG (mg/mL) and serum total protein (g/dL) concentrations and to estimate the apparent efficiency of absorption of IgG (%). The 129 heifers calves enrolled (CON=60; GAC=69) were also followed until weaning to assess the effect of GAC addition on preweaning health and growth. Multivariable linear regression showed that the addition of GAC to CR did not influence passive transfer of IgG, as measured by apparent efficiency of absorption at 24h of age (CON=54.0 vs. GAC=54.3%), serum IgG (CON=20.3 vs. GAC=20.2mg/mL), and serum total protein (CON=5.69 vs. GAC=5.68g/dL). Although study sample sizes were not originally derived to evaluate health outcomes, treatment had no effect on weight gain or incidence of health events (diarrhea, pneumonia, mortality) for heifer calves between birth and 7 wk of age.

When and where do dairy cows defecate and urinate?

The accumulation of urine and feces can be responsible for many cow and environmental problems. Despite this, little is known about the factors affecting defecation and urination. In the first experiment, the occurrence of defecation and urination behaviors of 48 lactating Holstein cows was observed [days in milk (DIM) = 144.7 ± 38.0 d, body weight (BW) = 667.1 ± 72.0 kg, parity = 2.8 ± 2.3] in freestalls over 48 h. In the second experiment, defecation and urination by 29 lactating Holstein dairy cows were observed (DIM = 62 ± 22.1 d, BW = 590 ± 70.0 kg, parity = 2 ± 1.3) in another freestall barn over a period of 5 d and related to cow activity and feeding behavior. In both experiments, based on total occurrence of eliminative behaviors, cows mainly defecated (experiment 1: 33.4 ± 2.0%; experiment 2: 42.3 ± 3.1%) and urinated (experiment 1: 28.2 ± 2.5%; experiment 2: 42.7 ± 4.0%) in the feed alley and while occupying a stall (defecation: experiment 1: 28.5 ± 1.0%; experiment 2: 26.2 ± 3.0%; urination: experiment 1: 42.2 ± 1.5%; experiment 2: 39.9 ± 3.8%). Occupying a stall included lying, standing in the stall, or standing with 2 feet in the stall and 2 feet in the alley. In both experiments, differences were found between cows in frequency of defecation (experiment 1: 9.8 ± 4.2/d, range = 3 to 20; experiment 2: 15.4 ± 4.3/d, range = 6 to 36) and in frequency of urination (experiment 1: 7.0 ± 3.1/d, range = 2 to 18; experiment 2: 9.3 ± 2.8/d, range = 3 to 19). Large differences between cows were observed in the frequency of defecation and urination, but these were not correlated with parity, milk production, BW, DIM, or dry matter intake.

Oestrogen and testosterone modulate the firing activity of dorsal raphe nucleus serotonergic neurones in both male and female rats.

Women are twice as likely to suffer from mood disorders than men. Moreover, a growing body of evidence suggests a reciprocal modulation between sex steroids and the serotonin (5-HT) system. A previous study from our laboratory has shown that the progesterone metabolites 5beta-pregnane-3,20-dione (5beta-DHP) and 5alpha-pregnan-3alpha-ol,20-one (3alpha,5alpha-THP), as well as dehydroepiandrosterone (DHEA), increase the firing activity of dorsal raphe nucleus (DRN) 5-HT neurones in female rats. The present study was undertaken to assess the effects of these steroids in male rats, as well as the effects of testosterone and 17beta-oestradiol (17beta-E) in both sexes, and finally to evaluate gender differences in the modulation of the 5-HT neuronal firing activity by these different neuroactive steroids. Male rats were treated i.c.v., for 7 days, with a dose of 50 microg/kg/day of one of the following steroids: progesterone, 5beta-DHP, 3alpha,5alpha-THP, DHEA, testosterone, 17beta-hydroxy-5alpha-androstan-3-one (5alpha-DHT) and 17beta-E. Some rats also received a 3-day administration of testosterone (50 microg/kg/day, i.c.v). Females were treated in the same fashion with testosterone and 17beta-E. Extracellular unitary recordings of 5-HT neurones, obtained in vivo in the DRN of these rats, revealed that testosterone and 17beta-E increased the firing activity of 5-HT neurones in both males and females. In males, the effect of testosterone could already be seen after 3 days of treatment. Neither castration nor any treatment with other steroids significantly modified the firing rate of male 5-HT neurones. Taken together with previous findings, the results of the present study indicate both similarities and differences between sexes in the modulation of 5-HT neurones by some steroids. This could prove important in understanding gender differences in mood disorders.

Modulation of the firing activity of female dorsal raphe nucleus serotonergic neurons by neuroactive steroids.

Important gender differences in mood disorders result in a greater susceptibility for women. Accumulating evidence suggests a reciprocal modulation between the 5-hydroxytryptamine (5-HT) system and neuroactive steroids. Previous data from our laboratory have shown that during pregnancy, the firing activity of 5-HT neurons increases in parallel with progesterone levels. This study was undertaken to evaluate the putative modulation of the 5-HT neuronal firing activity by different neurosteroids. Female rats received i.c.v. for 7 days a dose of 50 micro g/kg per day of one of the following steroids: progesterone, pregnenolone, 5beta-pregnane-3,20-dione (5beta-DHP), 5beta-pregnan-3alpha-ol,20-one, 5beta-pregnan-3beta-ol,20-one, 5alpha-pregnane-3,20-dione, 5alpha-pregnan-3alpha-ol,20-one (allopregnanolone, 3alpha,5alpha-THP), 5alpha-pregnane-3beta-ol,20-one and dehydroepiandrosterone (DHEA). 5beta-DHP and DHEA were also administered for 14 and 21 days (50 micro g/kg per day, i.c.v.) as well as concomitantly with the selective sigma 1 (sigma1) receptor antagonist NE-100. In vivo, extracellular unitary recording of 5-HT neurons performed in the dorsal raphe nucleus of these rats revealed that DHEA, 5beta-DHP and 3alpha,5alpha-THP significantly increased the firing activity of the 5-HT neurons. Interestingly, 5beta-DHP and DHEA showed different time-frames for their effects with 5beta-DHP having its greatest effect after 7 days to return to control values after 21 days, whereas DHEA demonstrated a sustained effect over the 21 day period. NE-100 prevented the effect of DHEA but not of 5beta-DHP, thus indicating that its sigma1 receptors mediate the effect of DHEA but not that of 5beta-DHP. In conclusion, our results offer a cellular basis for potential antidepressant effects of neurosteroids, which may prove important particularly for women with affective disorders.

Gender and gonadal status modulation of dorsal raphe nucleus serotonergic neurons. Part I: effects of gender and pregnancy.

Gender differences in susceptibility to affective disorders are well documented. The ovarian steroids, estrogen (E) and progesterone (P), may modulate the function of the serotonergic (5-HT) system, implicated in the etiology and treatment of affective disorders. We tested the hypothesis that ovarian steroid modulation of 5-HT function could result in a modification of the 5-HT neuronal firing activity. Extracellular unitary recordings of dorsal raphe nucleus 5-HT neurons were obtained in male rats and in female rats during natural E and P fluctuations. The average firing activity of 5-HT neurons was significantly higher in males (41%) than in freely cycling (CF) and in ovariectomized (OVX) females. During pregnancy, it increased gradually and by up to 136% on gestational day 17, then declined before parturition. In the postpartum period (PP), the firing rate decreased markedly compared to P17 but remained 63% higher than in CF. During pregnancy, the firing rate variations were closely correlated with P plasmatic levels. Finally no modification of the basal firing activity of locus coeruleus noradrenergic neurons was found in any group tested. Our results thus reveal a gender and pregnancy-dependent modulation of 5-HT firing rate that would impact 5-HT-mediated neurotransmission.

Establishment and comprehensive analysis of a new human transformed follicular lymphoma B cell line, Tat-1.

A spontaneously EBV transformed follicular lymphoma (FL) cell line, Tat-1, was established from the lymph node biopsy specimen of a patient with B cell FL, grade 1 in transformation to high grade disease. Tat-1 cells expressed lymphoid markers and developed tumor masses in immunodeficient mice. Bcl-2, Bcl-X(L), Bax and p53 protein expression was revealed by Western blotting. Flow cytometric analysis confirmed P-gp expression. Cytogenetically, the Tat-1 cell line showed identical chromosomal alterations to that of the initial biopsy specimen, among which the most notable were the t(14;18) typical of FL and additional abnormalities involving chromosomes 1, 8 and 13. Multicolor FISH analysis delineated all abnormalities, including a t(1p;8q), a der(8)(8q24::14q32::18q21) and a der(13)(13q32::8q24::14q32::18q21). Further FISH investigations using a locus-specific probe cocktail containing c-myc, IgH and bcl-2 revealed fusion of these three loci on the derivatives 8 and 13, in addition to the derivative 14 IgH/bcl-2 fusion and an extra copy of c-myc on derivative chromosome 1. These results demonstrate an additional example of the deregulation of bcl-2 and c-myc expression through recombination with a single IgH enhancer region. The unusual molecular features of the Tat-1 cell line render it a unique tool for studies focused on cytogenetic alterations, expression of multidrug resistance phenotype and expression of anti-apoptotic proteins in FL.