The 3Rs, Replacement, Reduction and Refinement, is a framework to ensure the ethical and justified use of animals in research. The implementation of refinements is required to alleviate and minimise the pain and suffering of animals in research. Public acceptability of animal use in research is contingent on satisfying ethical and legal obligations to provide pain relief along with humane endpoints. To fulfil this obligation, staff, researchers, veterinarians, and technicians must rapidly, accurately, efficiently and consistently identify, assess and act on signs of pain. This ability is paramount to uphold animal welfare, prevent undue suffering and mitigate possible negative impacts on research. Identification of pain may be based on indicators such as physiological, behavioural, or physical ones. Each has been used to develop different pain scoring systems with potential benefits and limitations in identifying and assessing pain. Grimace scores are a promising adjunctive behavioural technique in some mammalian species to identify and assess pain in research animals. The use of this method can be beneficial to animal welfare and research outcomes by identifying animals that may require alleviation of pain or humane intervention. This paper highlights the benefits, caveats, and potential applications of grimace scales.

This experiment assessed the efficacy of the cauterisation procedure with or without pain relief (injectable meloxicam) in mitigating the acute stress response to tail docking. Male piglets (n = 432) were allocated to the following treatments at 2-d post-farrowing: (1) no handling, (2) sham handling, (3) tail docked using clippers, (4) tail docked using a cauteriser, (5) meloxicam + clipper, and (6) meloxicam + cauteriser. Meloxicam treatments used Metacam® at 5 mg/mL injected i.m. 1 h prior to tail docking. Blood samples were collected at 15 and 30 min post-treatment and analysed for total plasma cortisol. Behaviours indicative of pain such as escape attempts, vocalisations and standing with head lowered were measured. The duration of vocalisations and frequency of escape attempts during treatment were greater in all tail docking treatments compared to the sham treatment. Piglets in the clipper treatment had higher (p < 0.05) cortisol concentrations at 30 min but not 15 min after treatment and stood for longer (p < 0.001) with head lowered in the first 60 min after treatment than those in the cauterisation treatment. Meloxicam reduced (p < 0.05) both the cortisol response at 30 min after tail docking with the clipper as well as the behavioural response in the first 60 min after tail docking with the clipper. In comparison to the sham treatment, cortisol concentrations at 15 min were higher in the two tail docking treatments whereas the tail docking treatments with meloxicam were similar to the sham handling treatment. In comparison to the sham handling treatment, cortisol concentrations at 30 min post-docking were higher (p < 0.05) only in the clipper treatment. While cauterisation appears to be less aversive than the clipper procedure, the administration of meloxicam did not mitigate the behavioural response during tail docking using either procedure, but reduced standing with head lowered in the first hour after docking for both methods. The commercial viability of administration of meloxicam requires consideration before it is recommended for use compared to cauterisation alone, as it requires additional handling of piglets and costs.

This experiment assessed the efficacy of the cauterisation procedure with or without pain relief (injectable meloxicam) in mitigating the acute stress response to tail docking. Male piglets (n = 432) were allocated to the following treatments at 2-d post-farrowing: (1) no handling, (2) sham handling, (3) tail docked using clippers, (4) tail docked using a cauteriser, (5) meloxicam + clipper, and (6) meloxicam + cauteriser. Meloxicam treatments used Metacam® at 5 mg/mL injected i.m. 1 h prior to tail docking. Blood samples were collected at 15 and 30 min post-treatment and analysed for total plasma cortisol. Behaviours indicative of pain such as escape attempts, vocalisations and standing with head lowered were measured. The duration of vocalisations and frequency of escape attempts during treatment were greater in all tail docking treatments compared to the sham treatment. Piglets in the clipper treatment had higher (p < 0.05) cortisol concentrations at 30 min but not 15 min after treatment and stood for longer (p < 0.001) with head lowered in the first 60 min after treatment than those in the cauterisation treatment. Meloxicam reduced (p < 0.05) both the cortisol response at 30 min after tail docking with the clipper as well as the behavioural response in the first 60 min after tail docking with the clipper. In comparison to the sham treatment, cortisol concentrations at 15 min were higher in the two tail docking treatments whereas the tail docking treatments with meloxicam were similar to the sham handling treatment. In comparison to the sham handling treatment, cortisol concentrations at 30 min post-docking were higher (p < 0.05) only in the clipper treatment. While cauterisation appears to be less aversive than the clipper procedure, the administration of meloxicam did not mitigate the behavioural response during tail docking using either procedure, but reduced standing with head lowered in the first hour after docking for both methods. The commercial viability of administration of meloxicam requires consideration before it is recommended for use compared to cauterisation alone, as it requires additional handling of piglets and costs.

BACKGROUND: Doramapimod, a p38 MAPK inhibitor, is a potent anti-inflammatory drug that decreases inflammatory cytokine production in equine whole blood in vitro. It may have benefits for treating systemic inflammation in horses. OBJECTIVE: To determine whether doramapimod is well tolerated when administered IV to horses, and whether it has anti-inflammatory effects in horses in a low-dose endotoxemia model. ANIMALS: Six Standardbred horses. METHODS: Tolerability study, followed by a blinded, randomized, placebo-controlled cross-over study. Horses were given doramapimod, and clinical and clinicopathological variables were monitored for 24 hours. Horses then were treated with doramapimod or placebo, followed by a low dose infusion of lipopolysaccharide (LPS). Clinical variables (heart rate, rectal temperature, noninvasive blood pressure), leukocyte count and tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) concentrations were measured at multiple time points until 6 hours post-LPS infusion. RESULTS: No adverse effects or clinicopathological changes were seen in the safety study. When treated with doramapimod as compared to placebo, horses had significantly lower heart rates (P = .03), rectal temperatures (P = .03), and cytokine concentrations (P = .03 for TNF-α and IL-1β), and a significantly higher white blood cell count (P = .03) after LPS infusion. CONCLUSIONS AND CLINICAL IMPORTANCE: Doramapimod has clinically relevant anti-inflammatory effects in horses, likely mediated by a decrease in leukocyte activation and decrease in the release of pro-inflammatory cytokines. To evaluate its potential as a novel treatment for systemic inflammatory response syndrome in horses, clinical trials will be necessary to determine its efficacy in naturally occurring disease.

BACKGROUND: Equine gastrointestinal nematodes (GINs) have been the subject of intermittent studies in Australia over the past few decades. However, comprehensive information on the epidemiology of equine GINs, the efficacy of available anthelmintic drugs and the prevalence of anthelmintic resistance (AR) in Australasia is lacking. Herein, we have systematically reviewed existing knowledge on the horse GINs recorded in Australia, and main aspects of their pathogeneses, epidemiology, diagnoses, treatment and control. METHODS: Six electronic databases were searched for publications on GINs of Australian horses that met our inclusion criteria for the systematic review. Subsets of publications were subjected to review epidemiology, diagnoses, pathogeneses, treatment and control of GINs of horses from Australia. RESULTS: A total of 51 articles published between 1950 to 2018 were included. The main GINs reported in Australian horses were cyathostomins (at least 28 species), Draschia megastoma, Habronema muscae, H. majus, Oxyuris equi, Parascaris equorum, Strongyloides westeri and Trichostrongylus axei across different climatic regions of Queensland, New South Wales, Victoria, and Western Australia. Nematodes are diagnosed based on the traditional McMaster egg counting technique, though molecular markers to characterise common GINs of equines were characterised in 1990s. The use of anthelmintic drugs remains the most widely-used strategy for controlling equine GIN parasites in Australia; however, the threshold of faecal egg count that should trigger treatment in horses, remains controversial. Furthermore, anthelmintic resistance within GIN population of horses is becoming a common problem in Australia. CONCLUSIONS: Although GINs infecting Australian horses have been the subject of occasional studies over the past few decades, the effective control of GIN infections is hampered by a generalised lack of knowledge in various disciplines of equine parasitology. Therefore, coordinated and focused research is required to fill our knowledge gaps in these areas to maximise equine health and minimise economic losses associated with the parasitic infections in Australia.

A cross‐sectional study was conducted to estimate the seroprevalence of bovine brucellosis in smallholder farms in seven regions in Pakistan, identify herd and individual level risk factors for seropositivity and assess the level of engagement of farmers with risk factors. In total, 1063 cattle and buffalo belonging to 420 herds in seven districts were sampled. The Rose Bengal test (RBT), indirect enzyme‐linked immunosorbent assay (I‐ELISA) and competitive enzyme‐linked immunosorbent assay (C‐ELISA) were used for the serological diagnosis of bovine brucellosis on all the serum samples. The associations between herd‐and animal‐level risk factors and seropositivity were investigated using logistic regression analyses. In addition, herd management practice scores, created to quantify the number of management practices undertaken that pose a risk for Brucella transmission, were calculated and compared between seropositive and negative herds within each district. Overall herd and animal prevalence were estimated to be 16.2% (95% CI, 13–20%) and 8.7% (95% CI, 7.2–10.6%), respectively, across all districts sampled. Herds with a history of last trimester abortion were found to be more likely to be positive than herds without such history (OR = 2.06, 95% CI, 1.09–3.89), providing validation of our findings and identifying that clinical disease is occurring in this region. It was also identified that herds with five to eight buffalo (OR = 3.80, 95% CI, 1.69–8.49), and those with more than eight buffalo (OR = 3.81, 95% CI, 1.51–9.58) were more likely to be positive for Brucella than those with less (one to two and three to four) buffaloes present in the herd. The presence of other domestic animals at the farm and purchasing animals in last year were found to have no association with seropositivity. The findings of this study support the need for the development of targeted intervention strategies specific to the disease status of each district.

Timely euthanasia is a fundamental part of safeguarding swine welfare by reducing suffering when compromised pigs are unable to recover. The quality and appropriateness of timely euthanasia rely mainly on the knowledge and experience of the individual caretaker but may also be affected by caretaker attitudes toward euthanasia. However, literature on caretaker attitudes toward swine euthanasia is lacking. This study investigated caretaker attitudes, perceived knowledge, and confidence in performing on-farm timely euthanasia. A total of 84 caretakers from eight swine farms (ranging in size from 1,300 to 7,000 sows) participated in a survey designed to investigate attitudes toward swine and swine euthanasia. Caretaker's ages ranged from 18 to 59 yr with an average work experience of 8.5 yr. The majority of participants worked in either farrowing or breeding units. Survey questions were designed to assess caretakers' attitudes and attributes (empathy affect, empathy attribution, attitudes toward pigs, feeling bad about euthanizing pigs, and assumptions about pigs' emotional capabilities), decision-making skills (confidence in identifying compromised pigs or relying on coworkers to make decisions), and euthanasia skillset (confidence in performing euthanasia, training, and perceived level of knowledge). Using cluster analysis to analyze survey answers, three distinct groups of caretakers were identified: 1) confident and empathetic; 2) Confident, knowledgeable, and detached; and 3) unconfident and lacking knowledge. The survey results showed that empathy attribution was strongly correlated with empathy affect (r = 0.571, P < 0.01) and that empathy affect and empathy attribution were higher in female caretakers compared with male caretakers (P < 0.05). A risk analysis that included previously identified clusters showed that females were more likely to be grouped among caretakers that were confident and empathetic (P = 0.04), and caretakers with more than 2 yr of swine experience were more likely to be grouped as confident and skilled (P = 0.01), while the unconfident and empathetic were more likely to have had less than 2 yr of experience (P = 0.04). This study provides important information about variability in caretaker experience as well as their attitudes toward pigs and timely euthanasia. Increased knowledge about swine caretaker attitudes may be used to implement training and euthanasia protocols to increase both human and pig welfare on farm.

This research studied whether floor feeding group-housed sows their daily allocation over multiple feed drops per day provides more equitable feeding opportunities in later drops. Over four time replicates, 275 sows were mixed into groups of 10 for both their first and second gestations (200 sows/gestation, 126 sows observed in both gestations). The feeding behavior of individual sows was recorded for 10 min following each of four feed drops per day (0730, 0900, 1100, 1500 h) on days 2, 9 and 51 post-mixing. The location of feeding sows (i.e., feeding in areas associated with high, reduced or little/no food availability) was also recorded. Sow aggressive behavior on day 2 was used to classify sows as dominant (D), subdominant (SD), or submissive (SM). Dominant sows spent the most time feeding in areas of high-food availability (gestation 1, p < 0.001; gestation 2, p = 0.023); SD sows fed more frequently than D sows from areas of reduced food availability (gestation 1, p = 0.001; gestation 2, p = 0.025); and SM sows performed more feeding behavior in areas of little/no food availability (gestation 1, p < 0.001; gestation 2, p < 0.001). These relationships did not change over feed drops or days in either gestation (p > 0.05). Further research on the management and design of floor feeding systems is required, with a particular emphasis on increasing accessibility to sows that avoid the feeding area.

BACKGROUND: Hepatic arteriovenous malformations (HAVMs) are rare congenital lesions consisting of multiple high-pressure arteries feeding into low-pressure veins via a central nidus. Massive haemorrhage, portal hypertension and hepatic insufficiency can ensue. Endovascular embolization is increasingly a first line treatment method although there is no general consensus or guidelines on the most effective embolic agent or approach. We describe the novel treatment of two dogs with congenital hepatic AVMs using a modified version of the 'pressure cooker' technique often utilised in neurointervention with the DMSO-based PHIL embolic agent delivered via the DMSO compatible Scepter-XC dual lumen balloon catheter. CASE PRESENTATION: Two paediatric dogs were diagnosed with hepatic AVMs. Both dogs presented with ascites and abnormal liver function tests. CT angiograms revealed hepatic arterio-portal malformations arising from an enlarged celiac artery. Selective catheterisation of the artery supplying the AVM was achieved via a femoral artery approach. A Scepter XC dual-lumen compliant balloon microcatheter and Traxcess 0.014 guidewire combination was advanced to the nidus via through the 5Fr guide catheter towards the nidus. Inflation of the balloon occluded arterial inflow and PHIL was injected under continuous fluoroscopic screening until the PHIL embolic agent penetrated into the draining portal vein beyond the nidus. In patient 1, normal portal venous waveform was restored with reversal of severe hepatic insufficiency. Whilst there was initial improvement post-operatively in patient 2 with normalisation of portal vein pressures and flow, opening of collateral nidus vessels re-established the high-pressure communication, and euthanasia was elected by the owner. CONCLUSIONS: The 'pressure cooker' technique is a safe and efficacious approach to the treatment of canine HAVMs. The novel use of PHIL and the Scepter XC balloon catheter has several advantages over conventional endovascular approaches. Translational application to human paediatric interventions for similar conditions where embolic and contrast agent volume constraints are similar can be considered.

Achieving and maintaining high standards of animal welfare is critical to the success of a modern zoo. Research has shown that an animal's welfare is highly dependent on how various individual animal factors (e.g., species traits, genetics, temperament and previous experience) interact with environmental features (e.g., social grouping, enclosure design and sensory environment). One prominent feature of the zoo environment is the presence of visitors. Visitor contact can be unpredictable and intense, particularly in terms of auditory and visual interaction. Depending on an animal's perception of this interaction, visitors can have either negative, neutral or positive impacts on zoo animal behaviour and welfare. This paper reviews the literature on the implications and potential opportunities of human-zoo animal interactions on animal behaviour and welfare, with the aim of stimulating interest, understanding and exploration of this important subject. The literature to date presents a mixed range of findings on the topic. It is possible this variation in the responses of zoo animals to visitors may be due to species-specific differences, the nature and intensity of the visitor interactions, enclosure design, and individual animal characteristics. Analysing these studies and better understanding animal preferences and motivations can provide insight into what animals find negatively and positively reinforcing in terms of visitor contact in a specific zoo setting. This understanding can then be applied to either safeguard welfare in cases where visitors can have a negative impact, or, conversely, it can be applied to highlight opportunities to encourage animal-visitor interaction in situations where animals experience positive emotions associated with visitor interaction.

We examined the effects of regulating the viewing proximity of visitors and the intensity of visitor behaviour on behaviours indicative of fear and stress physiology of 15 zoo-housed little penguins (Eudyptula minor). A 2 × 2 factorial fully randomised design was used to examine the effects of regulating: 1) the viewing proximity of visitors to enclosure, 'normal viewing distance' and 'increased viewing distance' (using a physical barrier set up 2 m away from the enclosure), and 2) the intensity of visitor behaviour, 'unregulated visitor behaviour' and 'regulated visitor behaviour' (using signage and uniformed personnel). In addition, a treatment consisting of closing the enclosure to visitors was included. Penguin behaviour, visitor numbers and visitor behaviour were recorded by CCTV video recordings and direct observations, respectively. Penguin faecal glucocorticoid metabolites were also analysed as a measure of stress physiology. We found that increased viewing distance reduced (p < 0.05) all visitor behaviours except for loud vocalisations and tactile contact with penguins. However, there were no direct effects of signage and uniformed personnel on visitor behaviour (p > 0.05). As the regulation of viewing proximity increased from a closed exhibit to an open exhibit with increased viewing distance, and then to an open exhibit with normal viewing distance, this increased the proportion of penguins huddling (p = 0.0011), vigilant (p = 0.0060) and retreating (p = 0.00013), and decreased the proportion of penguins within 1 m of the visitor viewing area (p = 0.00066), surface swimming (p = 0.00091) and preening in the water (p = 0.042). There were also limited effects of regulating visitor behaviour on penguin behaviour. No treatment effects were found on faecal glucocorticoid metabolites (p > 0.05). These results indicate that regulating visitor viewing proximity affects penguin behaviours indicative of fear and visitor behaviour. This suggests that close visitor contact can be fear-provoking for little penguins but increasing the distance between visitors and penguins can reduce fear responses of penguins by regulating both viewing proximity and visitor behaviour. However, it is unclear whether these changes in penguin behaviour are due to the increased separation between visitors and penguins and/or specific visitor behaviours associated with close viewing proximity to the enclosure, such as leaning over the enclosure or tactile contact with the pool, which are impeded when visitors are further away.