Search

We need validated, practical methods for pain assessment in piglets. Pain assessment can help researchers, veterinarians and industry professionals identify the need for analgesia or other pain management approaches when applying painful procedures, such as surgical castration. A pain assessment tool, the Piglet Grimace Scale (PGS), was previously validated in this context, but it is not widely applied. It is important that the PGS can be applied by a range of people, not just pain assessment experts. Our objective was to study the validity and reliability of PGS ratings applied by swine industry professionals and the general public, to assess its potential utility in non-research applications. To do so, we conducted an online Qualtrics survey in which, after completing a brief online training module and a practice test, 119 respondents were asked to rate 9 piglet images showing facial expressions immediately after surgical castration or sham-handling. Respondents were provided information on the castration treatment for each image and had continuous access to the scale throughout the survey. The survey also contained demographic questions. Industry respondents were recruited through networking, and participants from the general public were recruited through Amazon Mechanical Turk. Four trained experts scored each image, and these scores were averaged to produce gold standard scores. Intraclass correlations indicated strong internal consistency among experts, industry and public. ANOVA demonstrated scoring to be moderately comparable between groups. Campbell and Fiske’s Multi-Trait Multi-Method framework provided qualified support for the internal validity and reliability of the PGS scale, even applied by non-experts (industry and public). Both response groups were able to recognize pain in castrated piglets. However, public respondents attributed higher levels of pain to sham-handled piglets than industry respondents (2.83 vs. 2.35; p=0.047), and both response groups systematically overestimated pain experience compared to the experts, suggesting more training may be necessary before using the scale in a diagnostic capacity. Nevertheless, overall findings support wide applicability of PGS, even with minimal training, to improve awareness, recognition and monitoring of swine pain among veterinarians, industry professionals and even members of the public.

The total number of acquired melanocytic nevi on the skin is strongly correlated with melanoma risk. Here we report a meta-analysis of 11 nevus GWAS from Australia, Netherlands, United Kingdom, and United States, comprising a total of 52,506 phenotyped individuals. We confirm known loci including MTAP, PLA2G6, and IRF4, and detect novel SNPs at a genome-wide level of significance in KITLG, DOCK8, and a broad region of 9q32. In a bivariate analysis combining the nevus results with those from a recent melanoma GWAS meta-analysis (12,874 cases, 23,203 controls), SNPs near GPRC5A, CYP1B1, PPARGC1B, HDAC4, FAM208B and SYNE2 reached global significance, and other loci, including MIR146A and OBFC1, reached a suggestive level of significance. Overall, we conclude that most nevus genes affect melanoma risk (KITLG an exception), while many melanoma risk loci do not alter nevus count. For example, variants in TERC and OBFC1 affect both traits, but other telomere length maintenance genes seem to affect melanoma risk only. Our findings implicate multiple pathways in nevogenesis via genes we can show to be expressed under control of the MITF melanocytic cell lineage regulator.

The total number of acquired melanocytic nevi on the skin is strongly correlated with melanoma risk. Here we report a meta-analysis of 11 nevus GWAS from Australia, Netherlands, UK, and USA comprising 52,506 individuals. We confirm known loci including MTAP, PLA2G6, and IRF4, and detect novel SNPs in KITLG and a region of 9q32. In a bivariate analysis combining the nevus results with a recent melanoma GWAS meta-analysis (12,874 cases, 23,203 controls), SNPs near GPRC5A, CYP1B1, PPARGC1B, HDAC4, FAM208B, DOCK8, and SYNE2 reached global significance, and other loci, including MIR146A and OBFC1, reached a suggestive level. Overall, we conclude that most nevus genes affect melanoma risk (KITLG an exception), while many melanoma risk loci do not alter nevus count. For example, variants in TERC and OBFC1 affect both traits, but other telomere length maintenance genes seem to affect melanoma risk only. Our findings implicate multiple pathways in nevogenesis.

Genetic selection for improved performance has necessitated the frequent update of amino acid requirements to support this improvement. The first objective of this dissertation was to investigate the tryptophan requirement of laying hens in peak production and the lysine and sulfur amino acid requirements of broiler chicks under a phase feeding scenario using different models. Digestible tryptophan requirements were estimated to be 137 for egg production, 133 for egg mass, and 133 for feed efficiency using the linear broken-line model. The lysine and sulfur amino acid requirement were estimated by conducting 3 experiments within the starter phase from 2 to 5, 5 to 8, and 8 to 11 days of age. The linear broken line estimate for digestible lysine was 1.22, 1.17 and 1.16% for BWG and 1.31, 1.21, and 1.14% for FCR in experiments 1, 2, and 3, respectively. The linear broken line estimate for sulfur amino acids was 0.82, 0.81 and 0.94% for BWG and 0.82, 0.80, and 0.90 for FCR in experiments 4, 5, and 6, respectively. The lysine requirements decreased from 2 to 5 to 8 to 11 days, however the sulfur amino acid requirements increased during that same period. This could be due to other roles that sulfur amino acids play in the body other than growth. The second objective of this dissertation was to investigate the effects of a direct fed microbial (DFM) on broilers exposed to a cyclic heat stress in 2 consecutive experiments. In this experiment, the heat stress treatment reduced body weight gain and lean tissue accretion from 0 to 35 d in both experiments. In Experiment 2, when the litter was reused BWG was increased by 36 g/bird with supplementation of DFM. Ileal digestibility at 28 d (2 hr post HS) was improved with DFM supplementation in both experiments. Serum FITC-d increased with HS at both 28 and 35 d. Serum FITC-d was generally decreased with DFM at 28 d but the response was inconsistent at 35 d. Overall, the results suggest that HS reduced broiler performance and DFM treatment improved intestinal permeability and nutrient digestibility responses to HS in both experiments but did not improve performance until built up litter was used in Experiment 2. The last objective of this dissertation was to investigate the effects of sulfur amino acids (SAA) on broilers exposed to a cyclic heat stress. As expected, HS reduced BWG and worsened FCR. The supplementation of SAA had no effect on live performance. At 28 d of age, supplementation of SAA to birds exposed to HS resulted in reduced intestinal permeability. The interaction was lost at 31 d, but HS still increased intestinal permeability (P ≤ 0.05). Potential oxidative damage was reduced by increased SAA supplementation as indicated by an increase in the reduced glutathione to oxidized glutathione ratio. These data suggest that intestinal permeability is compromised acutely to at least three days of heat exposure before the bird can adjust, but oxidative damage is more chronic building over the entire 7 d HS period. SAA might have some protective effect on both intestinal permeability and oxidative stress responses to HS. Doctor of Philosophy To provide low-cost meat and eggs to consumers, the poultry industry must focus on improving efficiency as well as reduce the impact of stressors within the environment. This is achieved mostly by genetic selection which has resulted in significant improvements in both egg production in laying hens and body weight in broiler chickens. To support this improvement in growth rate birds would require a higher amino acid dense diet to support the increased protein output. One objective of this dissertation was to update the requirement of three key amino acids (tryptophan, lysine, and sulfur amino acids) in both laying hens and broilers for better formulation of diets. These amino acids that were investigated are essential in poultry diets and cannot be produced by the birds. The results from the tryptophan experiment indicated that laying hens need 137 mg of tryptophan per day to maintain maximum egg production. Results from the lysine and sulfur amino acid requirement experiment in broiler chickens indicated that broilers need 1.31% lysine and 0.90% sulfur amino acids to support maximal growth, but these requirements change over time and would need to be adjusted based on the broiler's age. Heat stress in the poultry industry is a major challenge which can affect the growth rate as well as the welfare of broiler chickens. The second objective of this dissertation was to investigate different strategies to ameliorate the effects of heat stress. Supplementing direct fed microbials or probiotics in broiler diets has been proposed as one of those strategies. An experiment was conducted to investigate the effect of the supplementation of a DFM on broiler chickens exposed to heat stress. The results indicated that the DFM was able to ameliorate the negative impacts of the heat stress on nutrient digestibility and intestinal permeability but did not improve the growth of the chickens. Another experiment was conducted to evaluate another strategy to ameliorate the effects of heat stress on broiler chickens which was the supplementation of sulfur amino acids. The sulfur amino acids are not only used for protein synthesis but have other physiological roles in the body that are important specifically during heat stress. Results from this experiment indicated that sulfur amino acids were able to ameliorate the negative effect of heat stress on intestinal permeability and oxidative stress but did not improve the performance of chickens subjected to the heat stress.

In commercial settings, broiler chickens and rainbow trout are housed in barren environments under high stocking densities, due to an emphasis on production efficiency. These monotonous housing conditions do not provide broilers or trout with the ability to perform functional, highly-motivated behaviors and increase their susceptibility to excessive anxiety and fear, resulting in negative affective states and poor animal welfare. Affective state (or emotional state) is a cumulative product of short-term life experiences, ranging from positive to negative. Because affective states are largely influenced by environmental condition, determining animal affective state can provide useful information on how to improve housing conditions in order to ensure positive experiences and good animal welfare. Cognitive processes are closely associated with affective state; a "cognitive bias" occurs when affective state influences aspects of cognition, such as judgement and attention. Animals in positive affective states make optimistically-biased decisions during ambiguous situations, judging the situation as if it will produce a positive outcome, and show less bias towards a perceived threat, responding in a less anxious and calm manner. Animals in negative affective states make pessimistically-biased decisions during ambiguous situations, judging the situation as if it will result in a negative outcome. Additionally, animals in negative affective states will bias their attention towards a perceived threat rather than alternative stimuli, responding in an anxious manner. Therefore, judgement and attention bias tests can be used to determine animal affective states. In Chapter 3, a judgement bias test was used to determine affective state of broiler chickens housed in either complex (perches, dust bath, pecking stones, and rotating enrichment objects) or barren (no enrichment) environments under either high or low stocking densities. Broilers housed in complex environments responded more optimistically during the judgement bias test than broilers from barren environments, indicating the former were in a positive affective state. Stocking density did not impact their responses in the judgement bias test, indicating that affective states were not impacted by that treatment. In Chapter 4, an attention bias test was used to determine level of anxiety and a tonic immobility test was used to determine fear in order to investigate affective state of broilers housed in the same conditions as described for Chapter 3. Broilers housed in complex environments were less anxious during the attention bias test than broilers from barren environments, indicating environmental complexity reduced anxiety in broilers. Stocking density did not impact anxiety. Broilers from high stocking density environments had shorter tonic immobility durations than broilers from low stocking density environments, suggesting the former were less fearful. Environmental complexity did not impact fearfulness. In Chapter 5, a judgement bias test was used to determine affective state of rainbow trout housed in either complex (shelter structure and artificial plants) or barren (no enrichment) tanks under either low or high stocking densities. Trout housed in high stocking density tanks responded optimistically during the judgement bias test, indicating they were in a more positive affective state compared to trout housed in low stocking density tanks. Environmental complexity did not impact their responses in the judgement bias test, indicating no effect of enrichments on affective states was found. These results indicate a beneficial relationship of a complex environment on broiler chicken affective state, observed through an optimistic judgement bias and reduced attention bias (anxiety) towards a perceived threat. Thus, providing a complex housing environment for broilers can improve their welfare and result in a positive affective state. Rainbow trout reared at the tested high density resulted in a positive affective state, although complexity did not benefit their welfare. Our results contribute much needed information on stocking densities to ensure fish welfare. Overall, environmental complexity, not stocking density, had a positive impact on broiler chicken affective states. Rainbow trout affective states were positively impacted by stocking density, but not environmental complexity. Master of Science Conventional housing of broiler chickens and rainbow trout (both raised for meat) causes concern for their welfare and affective states. Environmental conditions can greatly impact animals' affective states–their long-term emotional state, ranging from positive to negative. In barren environments at high stocking densities, broiler chickens and rainbow trout are prevented from showing normal behaviors and these conditions can compromise their affective state and welfare. By 'asking' chickens and trout whether the glass is half full or half empty, we can determine level of optimism or pessimism, and level of anxiety or calmness, therefore gaining a better understanding of their affective states. This can be done using a judgement bias test and attention bias test, where animal responses (optimism and anxiety) are recorded during ambiguous situations (judgement) and threatening situations (attention). Animals in positive affective states judge ambiguous situations optimistically (glass half full) and pay little attention towards perceived threats, while animals in negative affective states judge the same ambiguous situations pessimistically (glass half empty) and pay more attention towards perceived threats. In Chapter 3, responses to ambiguous situations were used to determine the affective state of broiler chickens housed in either enriched (perches, dust bath, pecking stones, rotating toys) or barren environments at either high or low stocking densities. Broiler chickens housed in enriched environments had an optimistic judgement bias of ambiguous situations (glass half full), suggesting they were in a more positive affective state compared to broilers housed in barren environments. Stocking density did not impact their level of optimism. In Chapter 4, responses to a perceived threat were used to determine level of anxiety and a tonic immobility test was used to determine fear of broilers housed under the same conditions as in Chapter 3. Broilers housed in enriched environments paid less attention to a perceived threat than broilers housed in barren environments, indicating the former were less anxious (glass half full) and in a positive affective state. Fear was not impacted by the tested enrichments, but birds kept under higher stocking densities did show reduced fear compared to birds in low-density environments. In Chapter 5, rainbow trout were housed in either enriched (shelter structure and artificial plants) or barren tanks at either high or low stocking densities. Affective state was evaluated through their responses to ambiguous situations. Trout housed in high stocking density environments had an optimistic judgement bias of ambiguous situations (glass half full), suggesting they were in a more positive affective state than trout housed in low stocking density environments. The enrichments did not impact their responses during the test, suggesting they did not impact fish optimism. These results indicate that an enriched environment improves broiler affective state and welfare compared to conventional housing conditions, the tested densities did not impact their welfare. Although an enriched environment did not positively impact responses of trout during ambiguous situations, our results show that housing rainbow trout in large groups results in a positive affective state and improved welfare status compared to housing trout in small groups. Overall, environmental enrichment, not stocking density, had a positive impact on broiler chicken affective states. Rainbow trout affective states were positively impacted by stocking density, but not environmental enrichment.