Trade-offs in the externalities of pig production are not inevitable.

Farming externalities are believed to co-vary negatively, yet trade-offs have rarely been quantified systematically. Here we present data from UK and Brazilian pig production systems representative of most commercial systems across the world ranging from 'intensive' indoor systems through to extensive free range, Organic and woodland systems to explore co-variation among four major externality costs. We found that no specific farming type was consistently associated with good performance across all domains. Generally, systems with low land use have low greenhouse gas emissions but high antimicrobial use and poor animal welfare, and vice versa. Some individual systems performed well in all domains but were not exclusive to any particular type of farming system. Our findings suggest that trade-offs may be avoidable if mitigation focuses on lowering impacts within system types rather than simply changing types of farming.

Changes in Stereotypies: Effects over Time and over Generations.

Stereotypies comprise a wide range of repeated and apparently functionless behaviors that develop in individuals whose neural condition or environment results in poor welfare. While stereotypies are an indicator of poor welfare at the time of occurrence, they may have various consequences. Environmental enrichment modifies causal factors and reduces the occurrence of stereotypies, providing evidence that stereotypies are an indicator of poor welfare. However, stereotypy occurrence and consequences change over time. Furthermore, there are complex direct and epigenetic effects when mother mammals that are kept in negative conditions do or do not show stereotypies. It is proposed that, when trying to deal with challenging situations, stereotypies might initially help animals to cope. After further time in the conditions, the performance of the stereotypy may impair brain function and change brain connections, neurophysiology and eventually neuroanatomy. It is possible that reported neuroanatomical changes are an effect of the stereotypy rather than a cause.

Positive and negative gestational handling influences placental traits and mother-offspring behavior in dairy goats.

Dairy animals are subjected to a number of potential stressors throughout their lives, including daily interactions with humans. The quality of these interactions may have direct consequences for the animal undergoing the experience, but if such events occur during gestation it may also affect the developing fetus. This study examined the effects of differential handling during mid-gestation in 40 twin-bearing Saanen×Toggenburg primiparous goats. Between days 80 and 115 of gestation (gestation=150 days), goats were subjected to aversive (AVS, n=13), gentle (GEN, n=13) or minimal (M, n=14) handling protocols for 10 minute periods twice daily. The control (M) group did not receive handling treatments and all goats received normal husbandry procedures outside treatment periods. Salivary cortisol measured during the treatment period was higher in AVS goats (mean cortisol (sem) in pg/µl: AVS: 176.7 (18.2), GEN: 119.6 (11.1), M: 126.5 (13.7); P=0.007). Data collection was focussed on mother-offspring behaviors 2h post-partum, placental morphology and colostrum quality. AVS goats were the only treatment group to suffer fetal loss (16% loss vs 0% in GEN and M, P=0.05). Treatment also influenced placental morphology with a tendency for fewer cotyledons evident in placentae from the aversive treatment (AVS: 87.9 (7.8), GEN: 107.1 (7.9), M: 112.1 (9.3), P=0.093), and significantly fewer medium sized cotyledons (AVS: 67.6 (7.8), GEN: 89.3 (6.4), M: 84.3 (5.4), P=0.042). GEN goats displayed more grooming and nosing behaviors towards their young during the first 2h post-partum (grooming: GEN: 89.3% (7.1), AVS: 72.6% (7.7), M: 63.4% (9.0), P=0.045; nosing frequency: GEN: 58.8 (12.5), AVS: 28.6 (11.1), M: 34.7 (6.5), P=0.021). There was an overall trend for kids from mothers experiencing the AVS treatment to take longer to stand, reach the udder and suck compared to kids from GEN and M treatment groups. Treatment significantly affected latency to perform play behavior, with kids from AVS goats taking on average 25 min longer to play for the first time than kids from GEN and M treatment groups (P<0.001). The results show that handling during gestation affects placental morphology, fetal survival and post-partum maternal behaviors, and influences kid behavioral development. Such results have important animal welfare implications, demonstrating that negative handling of pregnant females results in poorer placental quality with potential for fetal loss. It also demonstrates the beneficial effects of positive handling on enhancement of maternal behaviors.

Simultaneous detection and quantification of six equine cytokines in plasma using a fluorescent microsphere immunoassay (FMIA).

Cytokines are cell signalling proteins that mediate a number of different physiological responses. They are also biomarkers for inflammatory conditions and potential diagnostic references for diseases. Until recently, simultaneous quantification of cytokine profiles had not been possible. Now however, fluorescent microsphere immunoassays (FMIA) are able to measure multiple cytokines in a single sample. The following pro-inflammatory and anti-inflammatory cytokines were quantified in equine plasma and serum samples: interleukin (IL)-2, IL-4, IL-6, IL-10, interferon (IFN)-γ, tumor necrosis factor (TNF)-α. •The objective of this study was to quantify six equine cytokines simultaneously using the BioPlex(®) 200 system in equine EDTA-plasma and serum.•It demonstrates an increased number of detectable cytokines over published studies.•This technology has the advantage of reduced sample volume and assay time compared to traditional sandwich ELISAs.

Early adverse experience alters dendritic spine density and gene expression in prefrontal cortex and hippocampus in lambs.

In the laboratory, prenatal stress produces alterations in the structure and function of corticolimbic neurons. Here we report changes in gene expression and corticolimbic dendritic spine morphology in the offspring of pregnant ewes subjected to aversive interactions with human handlers during the last five weeks of pregnancy (AVS) compared to control dams that received gentle handling (GEN). AVS lambs had higher spine density on pyramidal neurons in area CA1 of the hippocampus and in medial prefrontal cortex compared to GEN lambs, as well as a lower ratio of mushroom spines to stubby and thin spines in area CA1. Expression of genes involved in brain development and spine morphogenesis was decreased in hippocampus and prefrontal cortex in AVS compared to GEN lambs. This study is the first demonstration that an ecologically relevant aversive experience in a field setting alters neuronal structure similarly to previous reports from laboratory settings and that even for animals domesticated over 12,000 years ago, an apparently mild stressor, resulting from human-animal interactions, can have similarly profound impacts on corticolimbic morphology.

Identification of chromosomal locations associated with tail biting and being a victim of tail-biting behaviour in the domestic pig (Sus scrofa domesticus).

The objective of this study was to identify loci associated with tail biting or being a victim of tail biting in Norwegian crossbred pigs using a genome-wide association study with PLINK case-control analysis. DNA was extracted from hair or blood samples collected from 98 trios of crossbred pigs located across Norway. Each trio came from the same pen and consisted of one pig observed to initiate tail biting, one pig which was the victim of tail biting and a control pig which was not involved in either behaviour. DNA was genotyped using the Illumina PorcineSNP60 BeadChip whole-genome single-nucleotide polymorphism (SNP) assay. After quality assurance filtering, 53,952 SNPs remained comprising 74 animals (37 pairs) for the tail biter versus control comparison and 53,419 SNPs remained comprising 80 animals (40 pairs) for the victim of tail biting versus control comparison. An association with being a tail biter was observed on Sus scrofa chromosome 16 (SSC16; p = 1.6 × 10(-5)) and an unassigned chromosome (p = 3.9 × 10(-5)). An association with being the victim of tail biting was observed on Sus scrofa chromosomes 1 (SSC1; p = 4.7 × 10(-5)), 9 (SSC9; p = 3.9 × 10(-5)), 18 (SSC18; p = 7 × 10(-5) for 9,602,511 bp, p = 3.4 × 10(-5) for 9,653,881 bp and p = 5.3 × 10(-5) for 29,577,783 bp) and an unassigned chromosome (p = 6.1 × 10(-5)). An r(2) = 0.96 and a D' = 1 between the two SNPs at 9 Mb on SSC18 indicated extremely high linkage disequilibrium, suggesting that these two markers represent a single locus. These results provide evidence of a moderate genetic association between the propensity to participate in tail-biting behaviour and the likelihood of becoming a victim of this behaviour.

Factors affecting mechanical (nociceptive) thresholds in piglets.

OBJECTIVE: To evaluate the stability and repeatability of measures of mechanical (nociceptive) thresholds in piglets and to examine potentially confounding factors when using a hand held algometer. STUDY DESIGN: Descriptive, prospective cohort. ANIMALS: Forty-four piglets from four litters, weighing 4.6 ± 1.0 kg (mean ± SD) at 2 weeks of age. METHODS: Mechanical thresholds were measured twice on each of 2 days during the first and second week of life. Data were analyzed using a repeated measures design to test the effects of behavior prior to testing, sex, week, day within week, and repetition within day. The effect of body weight and the interaction between piglet weight and behaviour were also tested. Piglet was entered into the model as a random effect as an additional test of repeatability. The effect of repeated testing was used to test the stability of measures. Pearson correlations between repeated measures were used to test the repeatability of measures. Variance component analysis was used to describe the variability in the data. RESULTS: Variance component analysis indicated that piglet explained only 17% of the variance in the data. All variables in the model (behaviour prior to testing, sex, week, day within week, repetition within day, body weight, the interaction between body weight and behaviour, piglet identity) except sex had a significant effect (p < 0.04 for all). Correlations between repeated measures increased from the first to the second week. CONCLUSIONS AND CLINICAL RELEVANCE: Repeatability was acceptable only during the second week of testing and measures changed with repeated testing and increased with increasing piglet weight, indicating that time (age) and animal body weight should be taken into account when measuring mechanical (nociceptive) thresholds in piglets. Mechanical (nociceptive) thresholds can be used both for testing the efficacy of anaesthetics and analgesics, and for assessing hyperalgesia in chronic pain states in research and clinical settings.

The effect of biting tails and having tails bitten in pigs.

Tail-biting is a behavioral abnormality which compromises the welfare of pigs. The goal of this study was to characterize the tail-biting phenotype using behavior and measures of heart-rate (HR) and its variability (HRV) in pigs. Thirty pigs were categorized as tail-biters (n=10), tail-bite victims (n=10), and control pigs (n=10) based on the frequency of tail-biting behavior that they performed or received at the farm. The animals' behavioral responses were registered at the experimental facilities for 10 min during test sessions whereas physiological responses were registered for 10 min prior to (basal) and during sessions when subjected to a novel object test (NOT) and to a novel arena test (NAT). Phenotypes differed in most behaviors during the two tests and in the NOT their physiological responses suggested different regulation of vagal tone. Biters had a reduction from baseline values to values during testing for the root mean square of successive R-R intervals (RMSSD) and the high-frequency band (HF) compared to victims, whose RMSSD and HF increased from baseline to test values. In the low-frequency band (LF), an increase was shown in biters and controls while a decrease in victims. LF was found to be strongly positively correlated with HF and RMSSD in biters. During baseline, victims tended to have lower HF and significantly higher power of the low-frequency component divided by power of the high-frequency band (LF:HF ratio) compared to biters and controls. The activity of the autonomic nervous system, especially the suppression of parasympathetic tone, indicated that both victims and biters may have a dysfunctional autonomic regulation which may indicate psychological disturbance. We provide the first documentation of phenotypic differences between pigs that have performed tail-biting, have been victimized, or have not been involved in tail biting using HRV data.

Adults may be used to alleviate weaning stress in domestic foals (Equus caballus).

The present study aims to investigate whether the presence of unrelated adult horses at weaning would reduce the social stress of weaning and the emergence of undesirable behaviours. We tested this hypothesis in 32 domestic foals by comparing short and medium term behavioural and physiological responses to weaning in foals maintained in homogeneous groups of peers (PW) to those of foals grouped with both peers and unrelated adults (AW). In total, three trials were conducted, which each trial consisting of one AW group and one PW group. In all foals, weaning was followed by increased vocalization, increased locomotion and increased salivary cortisol concentration. However, signs of stress were less pronounced and shorter in duration in weanlings housed with unrelated adults (e.g., whinnies: p<0.05; salivary cortisol: p<0.05). Only foals without adults exhibited increased aggressiveness towards peers (p<0.05) and abnormal behaviours (p<0.05) such as excessive wood-chewing and redirected sucking towards peers. In conclusion, introducing adults to minimize weaning stress in foals and later on aggressiveness and abnormal behaviours appears as the most promising approach to date.

Effects of haloperidol, a dopamine D2-like receptor antagonist, on reward-related behaviors in laying hens.

In order to investigate the involvement of dopaminergic transmission in the regulation of reward-related behaviors in laying hens, the effects of systemic injections of dopamine D2-like receptor antagonist haloperidol (0.3, 0.5, 1.0 and 2.0 mg/kg s.c.) 30 min prior to a conditioned cue signaling a reward were tested and compared to the effects of a saline injection. Head movements and latency to initiate display of anticipatory behavior were significantly affected by 0.3 and 0.5 mg/kg haloperidol, respectively. More hens were found lying down resting prior to the cue at 0.5 mg/kg and higher doses, and increased latency to peck at reward and shorter duration of anticipatory behavior was significant at 2.0 mg/kg. The findings are consistent with the involvement of dopamine in control of reward-related behaviors in laying hens. It is suggested that the lowest dose of haloperidol (0.3 mg/kg) affects reward-related behaviors, whereas the effects of higher doses of haloperidol could be confounded by sedative effects. A high inter- and intra observer agreement in the assessment of head movements together with their dopamine dependency further suggest that this behavior in classical conditioning paradigm represents an indicator of the state of the reward system in laying hens that can be assessed with good reliability.

Applying fractal analysis to heart rate time series of sheep experiencing pain.

The objective assessment of pain is difficult in animals and humans alike. Detrended fluctuation analysis (DFA) is a method which extracts "hidden" information from heart rate time series, and may offer a novel way of assessing the subjective experience associated with pain. The aim of this study was to investigate whether any fractal differences could be detected in heart rate time series of sheep due to the infliction of ischaemic pain. Heart rate variability (HRV) was recorded continuously in five ewes during treatment sequences of baseline, intervention and post-intervention for up to 60 min. Heart rate time series were subjected to a DFA, and the median of the scaling coefficients (alpha) was found to be alpha=1.10 for the baseline sequences, 1.01 for the intervention sequences and 1.00 for the post-intervention sequences. The complexity in the regulation of heartbeats decreased between baseline and intervention (p approximately 0.03) and baseline and post-intervention (p approximately 0.01), indicating reperfusion pain and nociceptive sensitization in the post-intervention sequence. Random time series based on Gaussian white noise were generated, with similar mean and variance to the HRV sequences. No difference was found between these series (p approximately 0.28), pointing to a true difference in complexity in the original data. We found no difference in the scaling coefficient alpha between the different treatments, possibly due to the small sample size or a fear induced sympathetic arousal during test day 1 confounding the results. The decrease in the scaling coefficient alpha may be due to sympathetic activation and vagal withdrawal. DFA of heart rate time series may be a useful method to evaluate the progressive shift of cardiac regulation toward sympathetic activation and vagal withdrawal produced by pain or negative emotional responses such as fear.

Exploring non-invasive methods to assess pain in sheep.

The aim of this study was to determine whether changes in eye temperature, measured using infrared thermography (IRT), and heart rate variability (HRV) can detect moderate levels of pain in sheep. Six ewes received the following treatments: 1) noxious ischaemic stimulus by application of a forelimb tourniquet (S), 2) noxious ischaemic stimulus and flunixin meglumine (S+F), and 3) flunixin meglumine (F). Maximum eye temperature, HRV, mechanical nociceptive threshold, blood pressure and behaviour were recorded for up to 60 min, including 15 min of baseline, 30 min during intervention and 15 min post-intervention. There was a tendency towards a decrease in the heart rate variability parameters RMSSD (the root mean square of successive R-R intervals) and SDNN (the standard deviation of all interbeat intervals) in treatment S compared to treatment F, and a significant increase in the same parameters between test day 1 and 3. A reduction in eye temperature was detected for all treatments during intervention, but no difference was found between S and F and S+N and F during intervention. The eye temperature decreased more in test day 2 and 3 compared to test day 1 during intervention. A significant reduction for both lip licking and vocalisation was observed between test day 1 and 3, and forward facing ears was the ear posture most frequently recorded in test day 1. We suggest that HRV is a sensitive, non-invasive method to assess mild to moderate pain in sheep, whereas IRT is a less sensitive method.

Gene expression profiling of peripheral mononuclear cells in lame dairy cows with foot lesions.

Lameness is a major health issue and likely the single most common cause of pain and discomfort in dairy cattle. Appropriate treatment is delayed or neglected due, in part, to lack of reliable detection. Assessment of cows with lameness is currently limited to subjective visual scoring systems based on locomotion and posture abnormalities. These systems are unreliable to detect lameness, and therefore, a large number of cows remain undiagnosed. The objective of this research was to search for potential biomarkers for lameness-associated painful inflammatory foot lesions in dairy cattle using microarray-based gene expression profiling of peripheral blood mononuclear cells (PBMC). BOTL5 microarrays spotted in duplicate with cDNA representing bovine immune response genes were interrogated with cDNA samples in an eight-array, balanced complete block design with dye swap. Samples from eight lame cows with inflammatory foot lesions and from eight sound cows were pair-matched by age, weight, days in lactation, and pregnancy status at time of PBMC collection and directly compared with each other on individual arrays. Statistical analysis of resulting fluorescence intensity data revealed 31 genes that were putatively differentially expressed in lame versus sound cows (P<0.05). Of these, BLASTn analysis and gene ontology information showed that 28 genes had high similarity or homology to known human and/or rodent genes. Validation of 15 of these genes known to be important in inflammation and pain was carried out using relative quantitative real-time RT-PCR, which confirmed the up-regulation of interleukin (IL)-2 (12.68+/-1.47-fold increase) and IL-10 (2.39+/-0.55-fold increase), matrix metalloproteinase-13 (MMP-13) (10.44+/-1.14-fold increase), and chemokine C-C motif receptor-5 (CCR5) (5.26+/-1.05-fold increase), in lame relative to sound cows (P< or =0.05). Similarly, granulocyte-macrophage colony-stimulating factor receptor alpha chain precursor (GM-CSF-R-alpha) (2.30+/-0.63-fold increase) and IL-4 (2.06+/-0.59-fold increase) showed a tendency (P=0.10) for up-regulation in lame compared to sound cows. PBMC co-expression of IL-2, MMP-13, CCR5 and IL-10, and potentially IL-4 and GM-CSF-R-alpha appears to be a promising, objective sign of lameness-related inflammatory foot lesions in dairy cattle. In conclusion, this study revealed potential biomarkers of the presence of foot lesions that could boost diagnostic accuracy of lameness and, ultimately, help identify animals in need of pain relief.

Assessing mid-trimester salivary cortisol levels across three consecutive days in pregnant women using an at-home collection protocol.

The Pregnancy Outcomes and Community Health (POUCH) Study enrolled women in their 15th to 27th week of pregnancy from 52 prenatal clinics located in five communities. After the study began, an at-home protocol to measure maternal stress was added, which included collection of urine and saliva twice a day (waking = AM, bedtime = PM) for three consecutive days, and completion of a daily diary. The at-home protocol was in place for 2852 POUCH participants at enrollment, and 81.3% (n = 2318) consented and returned samples and diary. For these analyses, salivary cortisol was analysed in a subset of 846 women who delivered at term. Day-to-day correlation coefficients for AM cortisol (0.45-0.55) exceeded those for PM cortisol (0.31-0.43). Study diaries indicated that there was variation in the time interval between waking and sample collection. Analyses of discrete intervals showed the same awakening response pattern in cortisol levels that has been reported in studies with serial AM sampling. The adjusted mean AM cortisol (microg/dL) was 0.506 at 0-15 min post-waking, 0.544 at 16-30 min (P < 0.05), 0.582 at 31-60 min (P < 0.01), and 0.515 at >60 min post-waking. In addition, adjusted mean AM cortisol in samples collected at or before 9 a.m. was higher than that in samples collected after 9 a.m. (0.564 vs. 0.510 microg/dL, P < 0.01). Among working women, adjusted mean AM cortisol was higher on work days than non-work days (0.564 vs. 0.489 microg/dL, P < 0.01), and in multiparae compared with primiparae (0.551 vs. 0.502 microg/dL, P = 0.07). The parity effect was not evident in non-working women. The adjusted mean PM cortisol significantly increased as week of pregnancy at sampling increased, but was not significantly related to time of collection, work day and parity. Factors influencing AM and PM cortisol levels in pregnancy appear to differ. In studies of AM cortisol levels, it is important to gather data on time of sample collection, interval from waking to sample collection, parity and work.

Results of a national survey of US veterinary college faculty regarding attitudes toward farm animal welfare.

OBJECTIVE: To examine attitudes toward farm animal welfare among veterinary college faculty. DESIGN: E-mail survey. STUDY POPULATION: 157 US veterinary college faculty with large animal or food animal emphasis. PROCEDURE: Veterinarians from 27 US veterinary colleges were contacted via e-mail and asked to complete a 7-page survey relating to farm animal welfare issues. Thirty-one percent of those contacted responded. RESULTS: 71% of respondents self-characterized their attitude toward farm animal welfare as "we can use animals for the greater human good but have an obligation to provide for the majority of the animals' physiologic and behavioral needs." An additional 19% of respondents were more concerned about animal welfare than was indicated by that statement, and 10% were less concerned about farm animal welfare than was indicated by that statement. Significant relationships among demographic variables and attitude scores were observed, including more concerned attitudes among females, those with more liberal political views, and those who cited lower religiosity. No relationship between attitude and age was observed. CONCLUSIONS AND CLINICAL RELEVANCE: Veterinary college faculty have the opportunity to impact many stakeholders within the animal agriculture industries (eg, future veterinarians and policy makers looking for a veterinary science perspective). Results indicated that a considerable level of concern toward farm animal welfare is present in this population. Although the process of change may not be rapid, it is likely that the influence of these respondents will factor heavily into enhancing farm animal welfare.

Integrating animal welfare into veterinary education: using an online, interactive course.

Veterinarians in the United States and abroad are faced with growing public concern for the welfare of animals, particularly those in production. To prepare veterinarians to exert the leadership expected by the public and industry, steps should be taken to provide instruction in animal welfare at veterinary colleges. The ultimate goal is to offer courses in animal welfare in a consistent manner on a global scale, utilizing existing expertise in an efficient and cost-effective manner. Given the intense curricula of veterinary schools and the scarcity of instructors trained in animal welfare, a nontraditional approach is needed to educate veterinary students in the United States and abroad in animal welfare. Michigan State University (MSU) is developing a graduate-level, online interactive course in animal welfare assessment. The course will approach the topic of animal welfare education from a holistic, multidisciplinary standpoint (encompassing ethics, economics, and behavior) and address issues important to the general public and the international community. The MSU course will draw on renowned international animal welfare experts, allowing students to receive high-quality education that would be difficult in any other circumstance. The course will bridge an important gap in the veterinary curriculum and offer a complete and congruous education in animal welfare to veterinarians worldwide. The MSU course will also serve as a model for collaboration in content assembly and course delivery, by using technology to leverage global expertise in the interests of educational equity. In addition to innovative technology, such as the use of Web-collaboration software to create the course, a variety of media that enable students to interact with the material will also be incorporated throughout the course. Thus, the course will not only utilize the Internet to provide access to high-quality material, but will also require the active participation of the student, which is needed for effective learning.

Development of a porcine brain cDNA library, EST database, and microarray resource.

Recent developments in expressed sequence tag (EST) and cDNA microarray technology have had a dramatic impact on the ability of scientists to study responses of thousands of genes to internal and external stimuli. In neurobiology, studies of the human brain have been expanding rapidly by use of functional genomics techniques. To enhance these studies and allow use of a porcine brain model, a normalized porcine brain cDNA library (PBL) has been generated and used as a base for EST discovery and microarray generation. In this report, we discuss initial sequence analysis of 965 clones from this resource. Our data revealed that library normalization successfully reduced the number of clones representing highly abundant cDNA species and overall clone redundancy. Cluster analysis revealed over 800 unique cDNA species representing a redundancy rate for the normalized library of 6.9% compared with 29.4% before normalization. Sequence information, BLAST results, and TIGR cluster matches for these ESTs are publicly available via a web-accessible database (http://nbfgc.msu.edu). A cDNA microarray was created using 877 unique porcine brain EST amplicons spotted in triplicate on glass slides. This microarray was assessed by performing a series of experiments designed to test hybridization efficiency and false-positive rate. Our results indicate that the PBL cDNA microarray is a robust tool for studies of brain gene expression using swine as a model system.