BACKGROUND: Dietary content and environmental factors can shape the gut microbiota, and consequently, the way the gut microbiota metabolizes fats, carbohydrates, and proteins, affecting overall health of the host. We evaluated the impact of 3 diets (all meat [raw], high-insoluble fiber dry extruded diet and hydrolyzed protein dry extruded diet) on the gut microbiota of healthy dogs in a cross-over sequential study. RESULTS: We showed that diet can have an effect on the gut microbiome in dogs, which was influenced by the order of feeding. High-protein (all meat) diets were characterized by an increase in bacteria belonging to the Fusobacteria and Bacteroidetes phyla, whereas a high-insoluble fiber commercial diet correlated with increases in Firmicutes and Actinobacteria phyla. However, the individual dog's baseline microbiota had the most impact on the magnitude and nature of the changes in response to dietary intervention. CONCLUSION: Our results suggest that the dog fecal microbiota is driven by protein and fiber composition to different degrees in individual animals, and targeted modification of these patterns could be useful in the modulation of the gut microbiota in different diseases.

Ethical food choices have become an important societal theme in post-industrial countries. Many consumers are particularly interested in the animal welfare implications of the various foods they may choose to consume. However, concepts in animal welfare are rapidly evolving towards consideration of all animals (including wildlife) in contemporary approaches such as "One Welfare". This approach requires recognition that negative impacts (harms) may be intentional and obvious (e.g., slaughter of livestock) but also include the under-appreciated indirect or unintentional harms that often impact wildlife (e.g., land clearing). This is especially true in the Anthropocene, where impacts on non-human life are almost ubiquitous across all human activities. We applied the "harms" model of animal welfare assessment to several common food production systems and provide a framework for assessing the breadth (not intensity) of harms imposed. We considered all harms caused to wild as well as domestic animals, both direct effects and indirect effects. We described 21 forms of harm and considered how they applied to 16 forms of food production. Our analysis suggests that all food production systems harm animals to some degree and that the majority of these harms affect wildlife, not livestock. We conclude that the food production systems likely to impose the greatest overall breadth of harms to animals are intensive animal agriculture industries (e.g., dairy) that rely on a secondary food production system (e.g., cropping), while harvesting of locally available wild plants, mushrooms or seaweed is likely to impose the least harms. We present this conceptual analysis as a resource for those who want to begin considering the complex animal welfare trade-offs involved in their food choices.

Increasing societal and customer pressure to provide animals with 'a life worth living' continues to apply pressure on livestock production industries to alleviate pain associated with husbandry practices, injury and illness. Over the past 15-20 years, there has been considerable research effort to understand and develop mitigation strategies for painful husbandry procedures in sheep, leading to the successful launch of analgesic approaches specific to sheep in a number of countries. However, even with multi-modal approaches to analgesia, using both local anaesthetic and non-steroidal anti-inflammatory drugs (NSAID), pain is not obliterated, and the challenge of pain mitigation and phasing out of painful husbandry practices remains. It is timely to review and reflect on progress to date in order to strategically focus on the most important challenges, and the avenues which offer the greatest potential to be incorporated into industry practice in a process of continuous improvement. A structured, systematic literature search was carried out, incorporating peer-reviewed scientific literature in the period 2000-2019. An enormous volume of research is underway, testament to the fact that we have not solved the pain and analgesia challenge for any species, including our own. This review has highlighted a number of potential areas for further research.

Surveys are used extensively in social research and, despite a lack of conclusive evidence of their 'representativeness,' probability internet panel (PIP) surveys are being increasingly used to make inferences about knowledge, attitude and behavior in the general population regarding a range of socially relevant issues. A large-scale survey of Australian public attitudes and behavior toward the red meat industry was undertaken. Samples were obtained using a random digit dialing telephone survey (Computer-Assisted Telephone Interviewing-CATI, n = 502 respondents) and a PIP survey (PANEL, n = 530 respondents) to examine differences between the two samples regarding attitudes and behavior relating to livestock use and welfare. There was little difference in demographics between the CATI and the PANEL surveys apart from highest level of education. However, there were differences between the two samples in both attitudes and behavior toward the red meat industry after controlling for education levels. The PANEL respondents gave generally more conservative responses than did the CATI respondents in the sense that they were more positive toward the livestock industries and animal welfare within these industries. Differences were also found between the respondents of the two samples regarding behavior that relates to the red meat industry, both community and consumer behavior. PANEL respondents were less engaged in community behaviors performed in opposition of the red meat industry when compared with the CATI sample. The majority of CATI and PANEL respondents were red meat eaters and there was no difference between respondents of the two samples in relation to red meat consumption, however, there were fewer vegetarians and vegans in the PANEL survey. Possible reasons for the observed differences are discussed, however, a definitive answer will depend on further research to identify the specific psychological factors that differ between samples derived from different survey methodologies.

BACKGROUND: The potential for an exotic disease incursion is a significant concern for the United Kingdom (UK) equine industry. Horse owners' perceptions of, and attitudes towards, exotic diseases can influence decisions to adopt disease preparedness strategies. The objectives of this study were to describe horse owners' 1) perceptions of the term 'exotic disease', and 2) attitudes towards their risk of being affected by an exotic disease. In order to address these objectives, qualitative content analysis was undertaken on data collected using two open-ended survey questions. RESULTS: Horse owners (n = 423) perceived exotic diseases as 1) belonging somewhere else, and 2) a dangerous threat to their horse(s). The term 'exotic' was associated with being foreign, non-native, and out-of-place in the UK. Attitudes towards exotic disease risk were summarised into four categories: 1) responsible horse owners prevent disease, 2) horse owners need support to stop disease spread, 3) risk depends on proximity to the 'risky' horse, and 4) some risk is inevitable. A 'responsible' owner was aware of health hazards and took actions to protect their horse from disease. Reliance on others, including stakeholders, to uphold disease prevention in the community led to feeling vulnerable to disease threats. When evaluating risk, horse owners considered which horses were the 'riskiest' to their horse's health (horses that travelled, participated in competitions, or were simply unfamiliar) and avoided situations where they could interact. Despite undertaking disease prevention measures, the perceived uncontrollable nature of exotic diseases led some owners to feel an incursion was inevitable. CONCLUSIONS: Without accounting for horse owners' perceptions of, and attitudes towards, exotic diseases, recommendations to increase preparedness may be ineffective. Improved communication among stakeholders in the industry may assist in clarifying expectations for exotic disease-specific prevention measures. A collaborative approach among horse owners and stakeholders is recommended to improve disease preparedness within the industry.

Equine herpesvirus myeloencephalitis (EHM) remains one of the most devastating manifestations of equine herpesvirus type 1 (EHV-1) infection but our understanding of its pathogenesis remains rudimentary, partly because of a lack of adequate experimental models. EHV-1 infection of the ocular vasculature may offer an alternative model as EHV-1-induced chorioretinopathy appears to occur in a significant number of horses, and the pathogenesis of EHM and ocular EHV-1 may be similar. To investigate the potential of ocular EHV-1 as a model for EHM, and to determine the frequency of ocular EHV-1, our goal was to study: (1) Dissemination of virus following acute infection, (2) Development and frequency of ocular lesions following infection, and (3) Utility of a GFP-expressing virus for localization of the virus in vivo. Viral antigen could be detected following acute infection in ocular tissues and the central nervous system (experiment 1). Furthermore, EHV-1 infection resulted in multifocal choroidal lesions in 90% (experiment 2) and 50% (experiment 3) of experimentally infected horses, however ocular lesions did not appear in vivo until between 3 weeks and 3 months post-infection. Taken together, the timing of the appearance of lesions and their ophthalmoscopic features suggest that their pathogenesis may involve ischemic injury to the chorioretina following viremic delivery of virus to the eye, mirroring the vascular events that result in EHM. In summary, we show that the frequency of ocular EHV-1 is 50-90% following experimental infection making this model attractive for testing future vaccines or therapeutics in an immunologically relevant age group.

BACKGROUND: In addition to probe sequence characteristics, noise in hybridization array data is thought to be influenced by competitive hybridization between probes tiled at high densities. Empirical evaluation of competitive hybridization and an estimation of what other non-sequence related features might affect noisy data is currently lacking. RESULTS: A high density array was designed to a 1.5 megabase region of the canine genome to explore the potential for probe competition to introduce noise. Multivariate assessment of the influence of probe, segment and design characteristics on hybridization intensity demonstrate that whilst increased density significantly depresses fluorescence intensities, this effect is largely consistent when an ultra high density offset is applied. Signal variation not attributable to sequence composition resulted from the reduction in competition when large inter-probe spacing was introduced due to long repetitive elements and when a lower density offset was applied. Tiling of probes immediately adjacent to various classes of repeat elements did not generate noise. Comparison of identical probe sets hybridized with DNA extracted from blood or saliva establishes salivary DNA as a source of noise. CONCLUSIONS: This analysis demonstrates the occurrence of competitive hybridization between oligonucleotide probes in high density tiling arrays. It supports that probe competition does not generate random noise when it is maintained across a region. To prevent the introduction of noise from this source, the degree of competition should be regulated by minimizing variation in density across the target region. This finding can make an important contribution to optimizing coverage whilst minimizing sources of noise in the design of high density tiling arrays.

BACKGROUND: Physical activity modulates expression of metabolic genes and may therefore be a prerequisite for metabolic responses to environmental stimuli. However, the extent to which exercise interacts with environmental conditions to modulate metabolism is unresolved. Hence, we tested the hypothesis that even low levels of physical activity are beneficial by improving metabolic responsiveness to temperatures below the thermal neutral zone, thereby increasing the capacity for substrate oxidation and energy expenditure. METHODOLOGY/PRINCIPAL FINDINGS: We used wild rats (Rattus fuscipes) to avoid potential effects of breeding on physiological phenotypes. Exercise acclimation (for 30 min/day on 5 days/week for 30 days at 60% of maximal performance) at 22°C increased mRNA concentrations of PGC1α, PPARδ, and NRF-1 in skeletal muscle and brown adipose tissue compared to sedentary animals. Lowering ambient temperature to 12°C caused further increases in relative expression of NRF-1 in skeletal muscle, and of PPARδ of brown adipose tissue. Surprisingly, relative expression of UCP1 increased only when both exercise and cold stimuli were present. Importantly, in sedentary animals cold acclimation (12°C) alone did not change any of the above variables. Similarly, cold alone did not increase maximum capacity for substrate oxidation in mitochondria (cytochrome c oxidase and citrate synthase activities) of either muscle or brown adipose tissue. Animals that exercised regularly had higher exercise induced metabolic rates in colder environments than sedentary rats, and temperature induced metabolic scope was greater in exercised rats. CONCLUSIONS/SIGNIFICANCE: Physical activity is a necessary prerequisite for the expression of transcriptional regulators that influence a broad range of physiological functions from energy metabolism to cardiovascular function and nutrient uptake. A sedentary lifestyle leads to decreased daily energy expenditure because of a lack of direct use of energy and a muted metabolic response to ambient temperature, which can be reversed even by low levels of physical activity.

BACKGROUND: Streptococcus suis is a major swine pathogen and zoonotic agent that mainly causes septicemia, meningitis, and endocarditis. It has recently been suggested that proteinases produced by S. suis (serotype 2) are potential virulence determinants. In the present study, we screened a S. suis mutant library created by the insertion of Tn917 transposon in order to isolate a mutant deficient in a cell surface proteinase. We characterized the gene and assessed the proteinase for its potential as a virulence factor. RESULTS: Two mutants (G6G and M3G) possessing a single Tn917 insertion were isolated. The affected gene coded for a protein (SSU0757) that shared a high degree of identity with Streptococccus thermophilus PrtS (95.9%) and, to a lesser extent, with Streptococcus agalactiae CspA (49.5%), which are cell surface serine proteinases. The SSU0757 protein had a calculated molecular mass of 169.6 kDa and contained the catalytic triad characteristic of subtilisin family proteinases: motif I (Asp200), motif II (His239), and motif III (Ser568). SSU0757 also had the Gram-positive cell wall anchoring motif (Leu-Pro-X-Thr-Gly) at the carboxy-terminus, which was followed by a hydrophobic domain. All the S. suis isolates tested, which belonged to different serotypes, possessed the gene encoding the SSU0757 protein. The two mutants devoid of subtilisin-like proteinase activity had longer generation times and were more susceptible to killing by whole blood than the wild-type parent strain P1/7. The virulence of the G6G and M3G mutants was compared to the wild-type strain in the CD1 mouse model. Significant differences in mortality rates were noted between the P1/7 group and the M3G and G6G groups (p < 0.001). CONCLUSION: In summary, we identified a gene coding for a cell surface subtilisin-like serine proteinase that is widely distributed in S. suis. Evidences were brought for the involvement of this proteinase in S. suis virulence.

BACKGROUND: Streptococcus suis is a zoonotic pathogen that infects pigs and can occasionally cause serious infections in humans. S. suis infections occur sporadically in human Europe and North America, but a recent major outbreak has been described in China with high levels of mortality. The mechanisms of S. suis pathogenesis in humans and pigs are poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: The sequencing of whole genomes of S. suis isolates provides opportunities to investigate the genetic basis of infection. Here we describe whole genome sequences of three S. suis strains from the same lineage: one from European pigs, and two from human cases from China and Vietnam. Comparative genomic analysis was used to investigate the variability of these strains. S. suis is phylogenetically distinct from other Streptococcus species for which genome sequences are currently available. Accordingly, approximately 40% of the approximately 2 Mb genome is unique in comparison to other Streptococcus species. Finer genomic comparisons within the species showed a high level of sequence conservation; virtually all of the genome is common to the S. suis strains. The only exceptions are three approximately 90 kb regions, present in the two isolates from humans, composed of integrative conjugative elements and transposons. Carried in these regions are coding sequences associated with drug resistance. In addition, small-scale sequence variation has generated pseudogenes in putative virulence and colonization factors. CONCLUSIONS/SIGNIFICANCE: The genomic inventories of genetically related S. suis strains, isolated from distinct hosts and diseases, exhibit high levels of conservation. However, the genomes provide evidence that horizontal gene transfer has contributed to the evolution of drug resistance.

Animals play a significant role in the production of film and television in Australia and globally. Given this, regulating and monitoring their welfare on- and off-set is imperative. We therefore aim to compare Australia’s state and territory-based legislation and regulation to those in the United States and the United Kingdom and assess regulations against the Five Domains Model of animal welfare. Historical examples of animal incidents in Australian film are used to illustrate potential deficiencies. We reviewed archived media for animal welfare incidents on and off production sets. We demonstrate a lack of uniformity, with 37.5% (3/8) of states and territories providing targeted Codes of Practice for animals in filmed media, and partially addressing behavioural interactions or mental state within the Five Domains Model. Three themes of welfare concerns were identified including incidents on-set, incidents off-set, and effects of portrayal on perception or ownership of specific species. This highlights the need for standardised national legislation and improved monitoring and regulation. Further research should quantify the number of animals used in productions, describe the type and duration of the work the animals undertake, investigate the frequency of animal welfare incidents, and explore alternative methods to the use of live animals in film and television.