Adult Ossabaw Pigs Prefer Fermented Sorghum Tea over Isocaloric Sweetened Water.

Ossabaw pigs (n = 11; 5-gilts, 6-barrows; age 15.6 ± 0.62 SD months) were exposed to a three-choice preference maze to evaluate preference for fermented sorghum teas (FSTs). After conditioning, pigs were exposed, in four sessions, to choices of white FST, sumac FST, and roasted sumac-FST. Then, pigs were exposed, in three sessions, to choices of deionized H2O (-control; avoidance), isocaloric control (+control; deionized H2O and sucrose), and blended FST (3Tea) (equal portions: white, sumac, and roasted sumac). When tea type was evaluated, no clear preference behaviors for tea type were observed (p > 0.10). When the 3Tea and controls were evaluated, pigs consumed minimal control (p < 0.01;18.0 ± 2.21% SEM), and they consumed great but similar volumes of +control and 3Tea (96.6 and 99.0 ± 2.21% SEM, respectively). Likewise, head-in-bowl duration was the least for -control, but 3Tea was the greatest (p < 0.01; 5.6 and 31.9 ± 1.87% SEM, respectively). Head-in-bowl duration for +control was less than 3Tea (p < 0.01; 27.6 vs. 31.9 ± 1.87% SEM). Exploration duration was the greatest in the area with the -control (p < 0.01; 7.1 ± 1.45% SEM), but 3Tea and +control exploration were not different from each other (1.4 and 3.0 ± 1.45% SEM, respectively). Regardless of tea type, adult pigs show preference for FST, even over +control. Adult pigs likely prefer the complexity of flavors, rather than the sweetness alone.

Evaluation of Precision Livestock Technology and Human Scoring of Nursery Pigs in a Controlled Immune Challenge Experiment.

The objectives were to determine the sensitivity, specificity, and cutoff values of a visual-based precision livestock technology (NUtrack), and determine the sensitivity and specificity of sickness score data collected with the live observation by trained human observers. At weaning, pigs (n = 192; gilts and barrows) were randomly assigned to one of twelve pens (16/pen) and treatments were randomly assigned to pens. Sham-pen pigs all received subcutaneous saline (3 mL). For LPS-pen pigs, all pigs received subcutaneous lipopolysaccharide (LPS; 300 µg/kg BW; E. coli O111:B4; in 3 mL of saline). For the last treatment, eight pigs were randomly assigned to receive LPS, and the other eight were sham (same methods as above; half-and-half pens). Human data from the day of the challenge presented high true positive and low false positive rates (88.5% sensitivity; 85.4% specificity; 0.871 Area Under Curve, AUC), however, these values declined when half-and-half pigs were scored (75% sensitivity; 65.5% specificity; 0.703 AUC). Precision technology measures had excellent AUC, sensitivity, and specificity for the first 72 h after treatment and AUC values were >0.970, regardless of pen treatment. These results indicate that precision technology has a greater potential for identifying pigs during a natural infectious disease event than trained professionals using timepoint sampling.

Development of a Minipig Model of BINT From Blast Exposure Using a Repeatable Mobile Shock Expansion Tube.

INTRODUCTION: The Office of Naval Research (ONR) sponsored the Blast Load Assessment Sense and Test (BLAST) program to provide an approach to operationally relevant monitoring and analysis of blast exposure for optimization of service member performance and health. Of critical importance in this effort was the development of a standardized methodology for preclinical large animal studies that can reliably produce outcome measures that cannot be measured in human studies to support science-based guidelines. The primary advantage of this approach is that, because animal studies report physiological measures that correlate with human neuropathology, these data can be used to evaluate potential risks to service members by accounting for the anatomical and physiological differences between humans and large animal models. This article describes the methodology used to generate a comprehensive outcome measure dataset correlated with controlled blast exposure. METHODS AND MATERIALS: To quantify outcomes associated with a single exposure to blast, 23 age- and weight-matched Yucatan minipigs were exposed to a single blast event generated by a large-bore, compressed gas shock tube. The peak pressure ranged from 280 to 525 kPa. After a post-exposure 72-hour observation period, the physiological response was quantified using a comprehensive set of neurological outcome measures that included neuroimaging, histology, and behavioral measures. Responses of the blast-exposed animals were compared to the sham-treated cohort to identify statistically significant and physiologically relevant differences between the two groups. RESULTS: Following a single exposure, the minipigs were assessed for structural, behavioral, and cellular changes for 3 days after exposure. The following neurological changes were observed: Structural-Using Diffusion Tensor Imaging, a statistically significant decrement (P < .001) in Fractional Anisotropy across the entire volume of the brain was observed when comparing the exposed group to the sham group. This finding indicates that alterations in brain tissue following exposure are not focused at a single location but instead a diffuse brain volume that can only be observed through a systematic examination of the neurological tissue. Cellular-The histopathology results from several large white matter tract locations showed varied cellular responses from six different stains. Using standard statistical methods, results from stains such as Fluoro-Jade C and cluster of differentiation 68 in the hippocampus showed significantly higher levels of neurodegeneration and increased microglia/macrophage activation in blast-exposed subjects. However, other stains also indicated increased response, demonstrating the need for multivariate analysis with a larger dataset. Behavioral-The behavior changes observed were typically transient; the animals' behavior returned to near baseline levels after a relatively short recovery period. Despite behavioral recovery, the presence of active neurodegenerative and inflammatory responses remained. CONCLUSIONS: The results of this study demonstrate that (1) a shock tube provides an effective tool for generating repeatable exposures in large animals and (2) exposure to blast overpressure can be correlated using a combination of imaging, behavioral, and histological analyses. This research demonstrates the importance of using multiple physiological indicators to track blast-induced changes in minipigs. The methodology and findings from this effort were central to developing machine-learning models to inform the development of blast exposure guidelines.

Evaluation of a novel computer vision-based livestock monitoring system to identify and track specific behaviors of individual nursery pigs within a group-housed environment.

Animal behavior is indicative of health status and changes in behavior can indicate health issues (i.e., illness, stress, or injury). Currently, human observation (HO) is the only method for detecting behavior changes that may indicate problems in group-housed pigs. While HO is effective, limitations exist. Limitations include HO being time consuming, HO obfuscates natural behaviors, and it is not possible to maintain continuous HO. To address these limitations, a computer vision platform (NUtrack) was developed to identify (ID) and continuously monitor specific behaviors of group-housed pigs on an individual basis. The objectives of this study were to evaluate the capabilities of the NUtrack system and evaluate changes in behavior patterns over time of group-housed nursery pigs. The NUtrack system was installed above four nursery pens to monitor the behavior of 28 newly weaned pigs during a 42-d nursery period. Pigs were stratified by sex, litter, and randomly assigned to one of two pens (14 pigs/pen) for the first 22 d. On day 23, pigs were split into four pens (7 pigs/pen). To evaluate the NUtrack system's capabilities, 800 video frames containing 11,200 individual observations were randomly selected across the nursery period. Each frame was visually evaluated to verify the NUtrack system's accuracy for ID and classification of behavior. The NUtrack system achieved an overall accuracy for ID of 95.6%. This accuracy for ID was 93.5% during the first 22 d and increased (P < 0.001) to 98.2% for the final 20 d. Of the ID errors, 72.2% were due to mislabeled ID and 27.8% were due to loss of ID. The NUtrack system classified lying, standing, walking, at the feeder (ATF), and at the waterer (ATW) behaviors accurately at a rate of 98.7%, 89.7%, 88.5%, 95.6%, and 79.9%, respectively. Behavior data indicated that the time budget for lying, standing, and walking in nursery pigs was 77.7% ± 1.6%, 8.5% ± 1.1%, and 2.9% ± 0.4%, respectively. In addition, behavior data indicated that nursery pigs spent 9.9% ± 1.7% and 1.0% ± 0.3% time ATF and ATW, respectively. Results suggest that the NUtrack system can detect, identify, maintain ID, and classify specific behavior of group-housed nursery pigs for the duration of the 42-d nursery period. Overall, results suggest that, with continued research, the NUtrack system may provide a viable real-time precision livestock tool with the ability to assist producers in monitoring behaviors and potential changes in the behavior of group-housed pigs.

Pre-Clinical Common Data Elements for Traumatic Brain Injury Research: Progress and Use Cases.

Traumatic brain injury (TBI) is an extremely complex condition due to heterogeneity in injury mechanism, underlying conditions, and secondary injury. Pre-clinical and clinical researchers face challenges with reproducibility that negatively impact translation and therapeutic development for improved TBI patient outcomes. To address this challenge, TBI Pre-clinical Working Groups expanded upon previous efforts and developed common data elements (CDEs) to describe the most frequently used experimental parameters. The working groups created 913 CDEs to describe study metadata, animal characteristics, animal history, injury models, and behavioral tests. Use cases applied a set of commonly used CDEs to address and evaluate the degree of missing data resulting from combining legacy data from different laboratories for two different outcome measures (Morris water maze [MWM]; RotorRod/Rotarod). Data were cleaned and harmonized to Form Structures containing the relevant CDEs and subjected to missing value analysis. For the MWM dataset (358 animals from five studies, 44 CDEs), 50% of the CDEs contained at least one missing value, while for the Rotarod dataset (97 animals from three studies, 48 CDEs), over 60% of CDEs contained at least one missing value. Overall, 35% of values were missing across the MWM dataset, and 33% of values were missing for the Rotarod dataset, demonstrating both the feasibility and the challenge of combining legacy datasets using CDEs. The CDEs and the associated forms created here are available to the broader pre-clinical research community to promote consistent and comprehensive data acquisition, as well as to facilitate data sharing and formation of data repositories. In addition to addressing the challenge of standardization in TBI pre-clinical studies, this effort is intended to bring attention to the discrepancies in assessment and outcome metrics among pre-clinical laboratories and ultimately accelerate translation to clinical research.

Noninvasive, In-pen Approach Test for Laboratory-housed Pigs.

Traumatic brain injury (TBI) incidences have increased in both civilian and military populations, and many researchers are adopting a porcine model for TBI. Unlike rodent models for TBI, there are few behavioral tests that have been standardized. A larger animal requires more invasive handling in test areas than rodents, which potentially adds stress and variation to the animals' responses. Here, the human approach test (HAT) is described, which was developed to be performed in front of laboratory pigs' home pen. It is noninvasive, but flexible enough that it allows for differences in housing set-ups. During the HAT, three behavioral ethograms were developed and then a formula was applied to create an approach index (AI). Results indicate that the HAT and its index, AI, are sensitive enough to detect mild and temporary alterations in pigs' behavior after a mild TBI (mTBI). In addition, although specific behavior outcomes are housing-dependent, the use of an AI reduces variation and allows for consistent measurements across laboratories. This test is reliable and valid; HAT can be used across many laboratories and for various types of porcine models of injury, sickness, and distress. This test was developed for an optimized manual timestamping method such that the observer consistently spends no more than 9 min on each sample.

Association of plasma haptoglobin concentration and other biomarkers with bovine respiratory disease status in pre-weaned dairy calves.

We conducted a nested, case-control study of pre-weaned dairy calves ( n = 477; 4 California dairy farms) to assess the association between bovine respiratory disease (BRD) and hematologic biomarkers, including plasma haptoglobin (Hp) and plasma bactericide (PB). At each location, heifer or bull dairy calves were observed 2-4 times per week until confirmed as BRD-positive using parallel interpretation of thoracic ultrasound examination and auscultation. In addition, control calves were enrolled after being confirmed as BRD-negative using ultrasound and auscultation. Complete blood counts (CBC), PB, and Hp concentrations were measured. Hp values were higher in calves with confirmed BRD than in controls ( p < 0.01). The area under the curve (AUC) for the various biomarkers was obtained from the corresponding receiver operating characteristic curves. The AUC for Hp was 0.68, a value greater than those for PB or the remaining CBC parameters, indicating that Hp may be the most useful biomarker of BRD in pre-weaned dairy calves. The cutoff value for Hp was 0.195 g/L.

Effect of oral administration of meloxicam prior to transport on inflammatory mediators and leukoctye function of cattle at feedlot arrival.

OBJECTIVE To investigate the effects of meloxicam administration before long-distance transport on inflammatory mediators and leukocyte function of cattle at feedlot arrival. ANIMALS 60 healthy yearling beef steers. PROCEDURES Single-source steers were assigned to a transported (n = 40) or nontransported (20) group. Then, half of the steers within each group were assigned to receive meloxicam (1 mg/kg, PO) or a lactose placebo (1 bolus/steer, PO). All steers were transported approximately 1,300 km overnight to a feedlot; however, the nontransported group was moved before treatment (meloxicam or placebo) administration and allowed a 17-day acclimation period, whereas the transported group was moved immediately after treatment administration on day -1. Blood samples for measurement of inflammatory mediators and leukocyte function were collected from all steers on days -1, 0, and 3. RESULTS For steers that received meloxicam, mean plasma meloxicam concentration for the transported group was significantly greater than that for the nontransported group on day 0. For steers that received the placebo, mean haptoglobin-matrix metalloproteinase-9 complex for the transported group was significantly greater than that for the nontransported group on day 0. Mean haptoglobin concentration, neutrophil L-selectin intensity, and polymorphonuclear leukocyte count for the transported group were significantly greater than those for the nontransported group. Mean substance P concentration for nontransported steers that received meloxicam was significantly lower than that for the other 3 treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated meloxicam administration to healthy steers immediately before long-distance transport did not significantly mitigate the effects of transport-induced stress on leukocyte function or inflammatory markers.

Stress, immunity, and the management of calves.

Despite many advances in management and housing of dairy calves, 1 in 10 US dairy heifers die before weaning. A better understanding of the internal and external stimuli that contribute to the physiological and behavioral responses of calves to stressors is needed to reduce the risk of morbidity and mortality. Feeding calves their first meal is crucial, as successful passive transfer reduces the risk of mortality and morbidity. Sexually dimorphic immune and stress responses appear to be present in young cattle, but more research is needed to determine if this is caused by human bias for female calves. After that first feeding, 1 in 10 heifers and most bull calves in the United States are transported to specialized calf-raising facilities, yet information is lacking on the newborn calf stress response during transit. Whether calves are raised on site or at a calf ranch, individual housing systems are commonly used in the United States to reduce the risk of pathogen exposure and provide individual feeding and healthcare. However, health, growth, and social implications may be present for calves in alternative systems with greater space allowance than conventional systems or group housing. Disbudding and castration are typically performed at an early age for dairy calves during the pre-wean stage. These stressors often take place when the calf has decreased passive transfer of Ig and immunity is developing. Availability of pain mitigation through anesthetics and analgesics is limited, but evidence indicates that analgesics attenuate suppressed leukocyte function during these procedures. Solid-feed intake is a primary measure for determining weaning readiness, but some milk replacer formulas may influence the calf's oral behaviors before weaning; therefore, alternate weaning methods may need to coincide with alternate milk replacer formulas. The calf's behavioral and stress response at weaning may influence its immunity during the transition from individual to group housing (commingling). Alternate commingling strategies and nutritional supplements may help with this transition, but more research is needed to explore feasible alternatives. Optimizing the calf's health and well-being at these early stages may improve its long-term health and welfare.

Survey of management practices related to bovine respiratory disease in preweaned calves on California dairies.

In the spring of 2013, a survey of California (CA) dairies was performed to characterize management practices related to bovine respiratory disease in preweaned calves, compare these practices across geographic regions of the state, and determine the principal components that explain the variability in management between herds. The questionnaire consisted of 53 questions divided into 6 sections to assess management practices affecting dairy calves from precalving to weaning. The questionnaire was mailed to 1,523 grade A licensed dairies in CA and 224 responses (14.7%) were collected. Survey response rates were similar over the 3 defined regions of CA: northern CA, northern San Joaquin Valley, and the greater southern CA region. The mean size of respondent herds was 1,423 milking cows. Most dairies reported raising preweaned calves on-site (59.7%). In 93.3% of dairies, preweaned calves were raised in some form of individual housing. Nonsaleable milk was the most frequent liquid diet fed to preweaned heifers (75.2%). Several important differences were identified between calf-raising practices in CA and practices reported in recent nationwide studies, including herd sizes, housing practices, and sources of milk fed to heifers. The differences between the CA and nationwide studies may be explained by differences in herd size. Regional differences within CA were also identified. Compared with the 2 other regions, northern CA dairies were found to have smaller herds, less Holstein cattle, calves remained with dams for longer periods of time after calving, were more likely to be certified organic dairies, and raised their own calves more often. Principal component analysis was performed and identified 11 components composed of 28 variables (questions) that explained 66.5% of the variability in the data. The identified components and questions will contribute to developing a risk assessment tool for bovine respiratory disease in preweaned dairy calves.

Sexually dimorphic innate immunological responses of pre-pubertal Brahman cattle following an intravenous lipopolysaccharide challenge.

This study was designed to characterize potential sexually dimorphic immunological responses following a lipopolysaccharide (LPS) challenge in beef cattle. Six female (heifers) and five male (bulls) Brahman calves (average age=253 ± 19.9 and 288 ± 47.9 days; average body weight=194 ± 11 kg and 247 ± 19 kg for heifers and bulls, respectively) were challenged with LPS (0.25 µg LPS/kg body weight). Following administration of LPS, all cattle displayed increased sickness behavior beginning at 0.5h, with heifers on average displaying less sickness behavior than bulls. A febrile response was observed in all animals following LPS administration, with a maximum response observed from 4 to 5.5h. The average rectal temperature response was greater in heifers than bulls. In all cattle there were elevated serum concentrations of cortisol from 0.5 to 8h, TNF-α from 1 to 2.5h, IL-6 from 2 to 8h, and IFN-γ from 2.5 to 7h after LPS challenge. Additionally, serum concentrations of TNF-α were greater in heifers than bulls from 1.5 to 2h after the LPS challenge. Concentrations of IFN-γ were also greater on average in bulls than heifers. Leukopenia occurred from 1 to 8h, with a decreased neutrophil to lymphocyte ratio for the first 5h among all calves. These data demonstrate the existence of a sexually dimorphic acute-phase response in pre-pubertal Brahman calves. Specifically, heifers may have a more robust acute response to LPS challenge, even though bulls display more signs of sickness.

Sexually dimorphic stress and pro-inflammatory cytokine responses to an intravenous corticotropin-releasing hormone challenge of Brahman cattle following transportation.

This study was designed to characterize potential sexually dimorphic stress and immunological responses following a corticotropin-releasing hormone (CRH) challenge in beef cattle. Six female (heifers) and six male (bulls) Brahman calves (264 ± 12 d of age) were administered CRH intravenously (0.5 µg of CRH/kg body mass) after which serum concentrations of cortisol increased from 0.5 h to 4 h. From 1 h to 4 h after CRH administration, serum cortisol concentrations were greater in heifers than in bulls. In all cattle, increased serum concentrations of TNF-α, IL-6 and IFN-γ were observed from 2.5 h to 3 h after CRH, with greater concentrations of IFN-γ and IL-6 in heifers than bulls. Heifer total leukocyte counts decreased 1 h after CRH administration, while bull leukocyte counts and percent neutrophils decreased 2 h after CRH administration. Heifers had greater rectal temperatures than bulls, yet rectal temperatures did not change following administration of CRH. There was no effect of CRH administration on heart rate. However, bulls tended to have increased heart rate 2 h after CRH administration than before CRH. Heifer heart rate was greater than bulls throughout the study. These data demonstrate that acute CRH administration can elicit a pro-inflammatory response, and cattle exhibit a sexually dimorphic pro-inflammatory cytokine and cortisol response to acute CRH administration.

Innate immune responses and health of individually reared Holstein calves after placement into transition-pens 23 d after weaning.

Sixty-four Holstein dairy bull calves were all reared in individual calf-hutches and pens until they were randomly assigned to treatments of Grouped (pens of 3, n=36 calves) or Control (left in home hutch, n=8 calves) at age 68±2·3 d (body weight 74·9±1·5 kg). Blood was drawn at age 66, 70, 74 and 88 d for ex-vivo immunological and biochemical analyses. Calf starter intake was measured daily and individual body weights were measured at age 68, 78, and 89 (±2·3 sd) d. Grouped-calves consumed less starter (P<0·05), and weighed 6·4±1·99 kg less (P<0·05) than Control-calves by age 89 d. Group housing was a mild stressor, as evident by a transient suppression in neutrophil oxidative burst at age 70 d, but there was a lack of difference in the neutrophil:lymphocyte ratio of peripheral leucocytes and neutrophil expression of L-selectin at age 70 and 74 d. However, grouped-calves had elevated total peripheral leukocyte counts at age 70 d (P<0·05) and tended (P<0·10) to be greater at age 74 and 88 d. In addition, neutrophil phagocytosis of Escherichia coli increased (P<0·05) at age 74 d in Grouped-calves. These data indicate that moving calves into transition-pens with 3 calves per group decreases performance, but this may not be due primarily to extreme stress or disease. These data do indicate that it is important that calves have a competent immune system and any potential stressors are limited when they are moved into transition-pens because they are exposed to a wider diversity and (or) load of pathogens.

Innate immune responses of temperamental and calm cattle after transportation.

The objective was to investigate measures of cellular innate immune responses among calm and temperamental Brahman bulls in response to handling and transportation. Sixteen Brahman bulls (344 ± 37 d of age; 271.6 ± 45.5 kg BW) classified as either calm (n=8) or temperamental (n=8) were loaded onto a trailer, transported for 4h to a novel facility, rested 16 h overnight, and then were returned to their original facility after a 4h transport. Blood samples were collected immediately prior to (time 0) and at 24, 48, and 96 h after initial loading for analyses of innate immune and blood parameters. Leukocyte counts did not differ (P>0.05) due to temperament before or after transportation, but neutrophil:mononuclear cell ratios were greater in temperamental bulls compared to calm bulls at 24h. At 24h, expression of peripheral neutrophil ß(2)-integrin decreased among all bulls compared with 0 h (P<0.01). Temperamental bulls had greater glucose and cortisol than calm bulls (P<0.01) at 48 h; whereas calm bulls had elevated neutrophil L-selectin expression, and phagocytic and oxidative burst activity compared with temperamental bulls (P<0.10) at 48 h. The supernatant collected from endotoxin-stimulated whole blood cultures had greater TNF-α concentrations at 48 h than at the other time points (P<0.05), but no temperament effect was observed (P>0.05). In contrast, 96 h after initial loading the supernatant TNF-α concentrations were lower (P<0.05) among all cattle. Lastly, transportation increased neutrophil phagocytosis, oxidative burst, and cell adhesion molecule expression 96 h post-transportation and the effect was more pronounced among calm bulls. These data suggest that neutrophils from calm bulls are more likely to resist microbial invasion at 96 h after transportation than neutrophils from temperamental bulls.

Temperament influences endotoxin-induced changes in rectal temperature, sickness behavior, and plasma epinephrine concentrations in bulls.

This study was designed to determine the influence of temperament on endotoxin-induced changes in body temperature, sickness behavior, and stress hormone concentrations in cattle. Brahman bulls were selected based on temperament score measured 28 d prior to weaning. In dwelling recording devices were used to monitor rectal temperature, and jugular catheters were used to collect blood samples to determine cortisol and epinephrine concentrations before and after LPS administration (0.5 µg/kg body weight). Temperamental bulls had the lowest peak rectal temperature and sickness behavior scores relative to the Calm and Intermediate bulls. Prior to the administration of LPS, Temperamental bulls had greater cortisol and epinephrine concentrations than Calm or Intermediate bulls. Cortisol concentrations increased following LPS administration but were not affected by temperament. Epinephrine concentrations peaked 1 h after LPS administration in Calm bulls. Temperamental bulls did not exhibit an epinephrine response to LPS challenge. These data demonstrate that the temperament of calves can modulate the physiological, behavioral, and endocrine responses of pre-pubertal Brahman bulls to endotoxin challenge. Specifically, temperament differentially affected the rectal temperature, sickness behavior and epinephrine, but not cortisol, responses to LPS challenge.