A history of FMD research and control programmes in Southeast Asia: lessons from the past informing the future.

Foot and mouth disease (FMD) is a major animal health problem within Southeast Asia (SEA). Although Indonesia and more recently the Philippines have achieved freedom from FMD, the disease remains endemic on continental SEA. Control of FMD within SEA would increase access to markets in more developed economies and reduce lost productivity in smallholder and emerging commercial farmer settings. However, despite many years of vaccination by individual countries, numerous factors have prevented the successful control of FMD within the region, including unregulated 'informal' transboundary movement of livestock and their products, difficulties implementing vaccination programmes, emergence of new virus topotypes and lineages, low-level technical capacity and biosecurity at national levels, limited farmer knowledge on FMD disease recognition, failure of timely outbreak reporting and response, and limitations in national and international FMD control programmes. This paper examines the published research of FMD in the SEA region, reviewing the history, virology, epidemiology and control programmes and identifies future opportunities for FMD research aimed at the eventual eradication of FMD from the region.

Innovative pain management solutions in animals may provide improved wound pain reduction during debridement in humans: An opinion informed by veterinary literature.

Painful animal husbandry procedures are routinely performed in a range of livestock species without analgesia. Recently, innovative strategies have been developed to address wound pain in these animals. In particular, a farmer-applied "spray and stay" approach that is administered directly to open wounds was developed (Tri-Solfen® Medical Ethics Pty Ltd., Melbourne, Victoria, Australia). This strategy anaesthetises the wounds immediately upon their formation, with long-lasting effect. This development, described as a "pain management revolution," has become firmly established in the Australian livestock industries and has global potential. The positive outcomes of this approach provide insights and highlight potential benefits that may be accrued from its use in human wound care, providing rapid-onset wound analgesia and/or anaesthetising wounds prior to cleansing and debridement procedures. If these benefits are realised from a clinician and patient perspective for wound debridement as an initial indication, it could provide new horizons in pain management for a spectrum of wound-related procedures. Evidence from use in animal husbandry does support the concept that multimodal anaesthesia holds great potential in the field of wound management across many procedures.

Investigating baseline red meat slaughter operator capacity and directions for development in Lao PDR.

A study of operator knowledge, attitudes and practices (KAP) in red meat slaughter premise operations in northern Laos was conducted and compared with international best practice, to inform future industry development. The survey interviewed 68 (of 94 possible participants) employees from all ten commercial slaughter premises in six districts in three northern Laos provinces. This was supported by observations of slaughter premises, processes and the conduct of personnel. Descriptive analysis and linear regression modelling identified significant KAP predictor factors, and a gap analysis supported or rejected inferences from the generally low KAP scores for human and animal health, animal welfare, good manufacturing practices (GMP), work conditions and economics. The median proportion of correctly/desirably answered knowledge-related questions was 35.2% (interquartile range [IQR] = 22.2-51.9%) with 27.3% (IQR = 15.9-31.8%) for the attitude-related questions and 21.4% (IQR = 14.3-35.7%) for the practice-related questions. Two districts had significantly lower KAP scores than other districts, and staff had the lowest and meat inspectors had the highest scores. This study indicates that the current KAP for red meat processing falls short of international standards and that training programmes on disease risks and prevention are important in facilitating red meat industry development.

Promoting transboundary animal disease risk management via a multiple health and husbandry intervention strategies in upland Lao PDR.

Surveys of smallholder farmer knowledge, attitude, practice (KAP) and income were conducted in 2011 and 2012 in five northern provinces of Lao People's Democratic Republic (Lao PDR; n = 200 farmers). Participating villages were classified as either from a livelihood development project (LV; n = 80 farmers) or a livestock research project that contained both 'high-intervention' (HI; n = 60 farmers) or 'low-intervention' (LI; n = 60 farmers) villages. Farmer training plus a suite of health and productivity interventions were implemented in HI sites; only vaccination was implemented in LI sites, whilst various interventions and limited vaccination were introduced to LV sites. Farmer interviews were conducted with survey questions on socioeconomic variables and KAP of large ruminant health and disease risk management, enabling determination of quantitative and dichotomous qualitative traits and comparison of results from HI, LI and LV villages. The average farmer income from sales of large ruminants in HI was USD 621, in LI was USD 547 and in LV was USD 225 (p < 0.001). The predicted mean of total knowledge scores (/42) in the 2012 survey in HI was 28, in LI was 22 and in LV was 17 (p < 0.001). We conclude that improved KAP of large ruminant health and production can be achieved by intensive training, although with some farmers yet to apply their knowledge on husbandry and biosecurity practices, continued learning support and closed linkage of research and development projects to improve extension capacity is recommended. This multiple participatory approach promoting biosecurity in addition to vaccination may provide a more sustainable pathway for the advancement of Lao PDR on the Progressive Control Pathway for foot and mouth disease control.

Prevalence and clinical impact of Toxocara vitulorum in cattle and buffalo calves in northern Lao PDR.

This study was completed to determine the prevalence and distribution of Toxocara vitulorum infection in cattle and buffalo calves and investigate its clinical impact in northern Lao PDR (Peoples Democratic Republic). The results aim to assist decisions on disease control measures that can contribute to increasing cattle and buffalo productivity within smallholder farming systems in tropical areas. A prevalence survey for T. vitulorum in buffalo and cattle calves aged <3 months was conducted between September 2009 and June 2010 in five provinces of northern Lao PDR using a two-stage sampling technique to select 69 villages and 899 calves, with faecal samples collected and examined for T. vitulorum eggs at a local laboratory. At the time of sampling, data on calf morbidity and anthelmintic treatment was also collected. Factors potentially associated with infection and severity of infection were analyzed at univariable and multivariable levels, using T. vitulorum status (positive/negative) and on the positive calves only, faecal egg count levels as outcome variables. The estimated prevalence of T. vitulorum in northern Lao was 22.6 % (95 % confidence interval [CI], 0.17-0.28), and 76.8 % of villages had at least one positive calf. Province was the only significant (p < 0.05) variable investigated associated with calf infection status. Species (buffalo) was the only variable significantly (p < 0.05) associated with higher egg per gram of faeces levels among infected calves. Prevalence in calves aged 1-21 days, the reported prepatent period, was 17.5 % (CI 0.11-0.24). Treatment levels were very low (8.2 %) and if treatment occurred it was mostly unsuccessful. The high and wide spread infection of T. vitulorum in cattle and buffalo calves identified in this survey is likely to result in suboptimal cattle and buffalo productivity. Improved management of T. vitulorum infection in cattle and buffalo calves in northern Lao PDR is indicated to reduce potential negative production impacts and enable more efficient development of large ruminant livestock industry as a pathway from rural poverty for smallholder farmers in northern Lao PDR. In addition to quantifying this disease problem in calves, the conduct of this applied participatory research study provided an important opportunity to improve animal health services by increasing the parasite, large ruminant handling and research knowledge and capacity of government animal health staff and farmers.

Significant mortality of large ruminants due to hypothermia in northern and central Lao PDR.

An extreme cold exposure event occurred between March 14th and 19th 2011 in northern and central Lao PDR resulting in a major mortality of cattle and buffalo. At least six northern and one central province reported losses, involving 46 districts and 1,384 smallholder farmers, with a total of 7,162 cattle and 3,744 buffalo reported to have died in association with cold weather. Affected animals were observed to shiver, display slow and shallow respiration, lose consciousness and eventually die. Many deaths occurred at night and were recorded in both sexes and all ages of large ruminants. However, mortalities occurred mostly in animals that were free-grazing in the forest and natural grassland, and exposed to the cold weather. Some housed animals that were provided with warmth from shelter and fires and supplementary feed did not die. Samples from dead animals collected for laboratory analysis confirmed that bacterial or viral pathogens were not present. The cause of the mortality was attributed to hypothermia, and the economic losses were estimated at USD 2,463,912.00. Xieng Khouang Province reported the most severe losses with deaths of 4,600 cattle and 1,665 buffalo. At Thong Haihin meteorological station in this province on March 16th and 17th 2011, minimum temperatures recorded were 6.7°C and 7.5°C and rainfall recorded was 36.6 mm and 61.7 mm, respectively. This was the first reported extreme cold event in living memory occurring between the end of dry season and beginning of the wet season in northern Laos. This event is reported in detail as it caused a major loss of wealth for poor smallholder farmers and indicates that strategies to minimise the impact of extreme cold weather events need to be included in livestock development extension programmes.

Anaemia in Lambs Caused by Mycoplasma ovis: Global and Australian Perspectives.

Mycoplasma ovis (formerly Eperythrozoon ovis) is a haemotropic parasitic bacterium found within erythrocytes and distributed widely in global sheep and goat production regions. M. ovis is transmitted by biting flies and by contaminated instruments, causing morbidity and mortalities from anaemia, usually within 6 weeks following blood-exposure procedures, particularly vaccination, castration, ear tagging, mulesing, and occasionally crutching and shearing. Affected animals develop haemolytic anaemia and may have jaundice, causing abattoir condemnations. The typical history, clinical and pathological findings, display of M. ovis in blood smears and/or by PCR is diagnostic, although immune responses deplete M. ovis from smears; hence, in-contact healthy animals should be examined. Differential diagnoses include haemonchosis, fasciolosis, malnutrition (copper or vitamin B12 deficiency), and plant toxicities. M. ovis parasitaemia may persist, with recrudescence following stressful events, although most older sheep remain immune. Human infections have been reported. Inadequate socioeconomic data present difficulties in assessing the impact of M. ovis on production and as antimicrobial therapy is ineffective, its control requires management practices that minimize the impact of invasive procedures in periods when risks of M. ovis transmission are more likely. Although considered an emerging pathogen, recent improvements in welfare attitudes and husbandry practices on Australian sheep farms may potentially limit the transmission of M. ovis.

Can improving animal health and biosecurity knowledge of para-veterinarians in Cambodia assist in addressing challenges in smallholder livestock farming?

Para-veterinarians are crucial in providing front-line veterinary care and advice to smallholder farmers in countries with under-resourced national veterinary services. Many of these individuals undergo basic training with minimal monitoring and evaluation of performance and knowledge levels. To identify the knowledge, attitudes and practices of Cambodian para-veterinarians a cross-sectional survey (n = 80) of Village Animal Health Workers (VAHWs) was conducted during August 2018 in the provinces of Tbong Khmum and Takeo in southern Cambodia. Descriptive statistics indicated that VAHWs had a high level (mean score 86%) of animal health and biosecurity knowledge. As the Royal Government of Cambodia continues to move towards commercial agricultural value chains, VAHWs are crucial to ensuring the safety and sustainability of these systems. Binomial logistic regression modelling found the length of employment (p = .003), reporting of pig diseases (p = .007), recommending the quarantine of newly purchased animals (p = .008), treatment of cases of haemorrhagic septicaemia with antibiotics (p = .032) and washing footwear with water when entering or leaving a farm (p = .008) were all positively associated with a higher (≥92%) animal health and biosecurity knowledge score. These results demonstrate the correlation of positive biosecurity and disease reporting behaviours with high levels of knowledge, providing evidence that improved training of VAHWs can ensure they remain important in supporting the under-resourced national veterinary services. Regular monitoring and evaluation of training programmes for VAHWs will likely further strengthen the livestock sector in Cambodia.

Efficacy and application of a novel topical anaesthetic wound formulation for treating cattle with Foot-and-Mouth disease: A field trial in Cameroon.

Recently, a wound dressing formulation, (Tri-Solfen®, Medical Ethics Pty Ltd, Australia; TS) registered for use in ruminant husbandry in Australia, was registered for Foot-and-Mouth Disease (FMD) therapy in large ruminants in Laos, following clinical observations of improved welfare and healing following treatment of FMD lesions. In November 2019, an FMD outbreak in Cameroon provided an opportunity for a field trial, comparing clinical responses and recoveries to treatments on a sample of cattle (n = 36) comprising three equal groups of animals (n = 12), comparing responses to three treatments: (i) the application to lesions of TS, (ii) the administration of parenteral oxytetraycline commonly used for FMD in Cameroon; and (iii) an untreated control group (C). Appetite scores, lesion healing scores, and changes in dimensions of lesions, were recorded over a 15-day study period. Cattle treated with TS achieved both superior appetite and lesion healing scores with more rapid reduction in dimensions of lesions than other groups. Farmer observations indicated the TS treatment group had a more rapid return to eating with cessation of excessive salivation, and more rapid return of mobility (walking) with absence of overt lameness. The findings indicate that although mortality is usually low in FMD outbreaks, the disease is a debilitating and painful disorder with negative animal welfare impacts that should be addressed. All farmers expressed their desire that the product be made available for use in their region and modelling indicates that TS therapy imposes no additional financial burden on farmers, with the treatment likely to be provided at a similar or reduced cost to current treatment choices. As use of antibiotics for treatment of a viral disease potentially increases pressures for development of antimicrobial resistance and residues in the food chain, TS as an alternative non-antimicrobial therapy should be promoted for wider use in FMD outbreaks.

In utero infection of cattle with Mycobacterium avium subsp. paratuberculosis: a critical review and meta-analysis.

Mycobacterium avium subsp. paratuberculosis (Mptb) causes Johne's disease in ruminants. Disease control programmes aim to break the faecal-oral cow-calf transmission cycle through hygienic calf rearing and removal of affected cows from the herd, but these programmes do not take account of the potential for congenital infection. The aims of this study were to critically review research on in utero infection, determine the prevalence of fetal infection in cattle through meta-analysis and estimate the incidence of calves infected via the in utero route. About 9% (95% confidence limits 6-14%) of fetuses from subclinically infected cows and 39% (20-60%) from clinically affected cows were infected with Mptb (P<0.001). These are underestimates for methodological reasons. The estimated incidence of calf infection derived via the in utero route depends on within-herd prevalence and the ratio of sub-clinical to clinical cases among infected cows. Assuming 80:20 for the latter, estimates of incidence were in the range 0.44-1.2 infected calves per 100 cows per annum in herds with within-herd prevalence of 5%, and 3.5-9.3 calves in herds with 40% prevalence. These estimates were not markedly sensitive to the value chosen for the proportion of clinical cases. In utero transmission of Mptb could retard the success of disease control programmes if the opportunities for post natal transmission via colostrum/milk and environmental contamination were able to be controlled. The consequences of fetal infection for the calves so infected are discussed in the context of diagnosis and vaccination together with recommendations for future research.

Financial impact of an outbreak of clinically diagnosed blackleg - a case study from Lao PDR.

Blackleg (BL) is an acute to peracute highly fatal infectious disease of mainly large ruminants characterised by lesions of myonecrosis caused by Clostridium chauvoei, usually presenting as a sudden onset of sporadic mortalities. In Southeast Asia, 'BL' is considered a cause of occasional outbreaks of a subacute febrile illness, although there are few published reports available. Investigation of a major outbreak of clinically diagnosed BL occurring in large ruminants (cattle and buffalo) in three neighbouring villages in central Laos in mid-2017, was conducted to determine the financial impacts of BL on smallholder livelihoods. Owners of BL-infected large ruminants in the three affected villages were interviewed (n = 27) and financial losses including losses due to mortality, morbidity and costs of treatments, were determined. The reports of clinical signs of subcutaneous swelling with palpable crepitus in febrile animals were considered consistent with a diagnosis of BL. The outbreak occurred in 47 of a total 449 households with large ruminants across the three villages, affecting 147 of a total population of 3505 'at risk' large ruminants with 71 deaths reported. At the household level, the mean morbidity and mortality rates were 5 ± 4(95% CI: 3-6) and 3 ± 2(95% CI: 1-4) heads per household, respectively. The estimated financial losses due to BL per affected household was USD 822 ± 692(95% CI: 518-1125), being 122% of their annual household income from large ruminant sales. The comparison between the estimated losses due to BL per village and cost of annual BL vaccination programmes, indicated a potential economic benefit of USD3.09 and USD12.37 for every dollar invested in the vaccination programme, if BL outbreaks occur every 20 and 5 years, respectively. This study indicates that clinically diagnosed BL can cause significant losses to smallholder households, and requires Lao animal health authorities to consider vaccination interventions to prevent losses from re-emergence of BL in the known endemically affected areas.

Peste des Petits Ruminants (PPR) virus serological surveillance in goats in Lao PDR: Issues for disease eradication in a low-resource disease-free setting.

Peste des Petits ruminants (PPR) is an economically important transboundary viral disease of goats. This study aimed to determine a baseline of serological evidence for Peste des petits ruminants virus (PPRV) in Lao goats. A total of 1,072 serum samples were collected by convenience sampling across five provinces in Laos and tested for antibody response to PPRV using a commercially available competitive ELISA. Positive antibody responses were found in 2.2% (95% CI 1.4, 3.2) of the samples. True prevalence calculations indicated a total overall sample prevalence of 1.7% (95% CI 0.9, 2.8). The highest provincial seroprevalences were Xiangkhouang (3.5%, 95% CI 1.6, 6.9) and Xayaboury (2.9% (95% CI 1.3, 5.7). There was no association between antibody response and each of the following factors: location, breed, gender or age. Considering the apparent absence of disease manifestation of PPR in Laos, likely explanations for the antibody positivity could include cross reaction to other Morbilliviruses such as Measles or Canine Distemper, importation of pre-vaccinated goats, need for test cut-off re-evaluation to be region specific, or a subclinical and a less virulent circulating virus. This study highlights that the sampled Lao goat population is highly likely to be naïve to PPRV and therefore at risk of an outbreak, possibly by transboundary incursion of livestock from PPR endemic China. Further work is required in the testing of small ruminants in Laos that may eventually provide evidence for a status of freedom from disease, particularly in support of programs aimed at global PPR eradication.

Risk factors for Neospora caninum, bovine viral diarrhoea virus, and Leptospira interrogans serovar Hardjo infection in smallholder cattle and buffalo in Lao PDR.

Smallholder large ruminant production in Lao People's Democratic Republic (Laos) is characterised by low reproductive efficiency. To determine if common abortifacient bovid infectious diseases are involved, a serological investigation was conducted. Sera was collected from stored and fresh cattle (n = 390) and buffalo (n = 130) samples from 2016-18 from, and then examined for associations in a retrospective risk factor study of 71 herds. The sera were assayed for antibodies to Neospora caninum, bovine viral diarrhoea virus (BVDV), Leptospira interrogans serovar Hardjo and Brucella abortus using commercially available enzyme-linked immunosorbent assay kits. These pathogens were detected in buffalo samples at 78.5% (95% CI 71.4-85.6), 0%, 2.3% (95% CI 0-4.9) and 0%, respectively, and in cattle at 4.4% (95% CI 2.4-6.4), 7.7% (95% CI 3.1-12.3), 12.8% (95% CI 9.5-16.1) and 0.26% (95% CI 0-0.8), respectively. Exposure of buffalo to N. caninum was positively associated with buffalo age, with a predicted seropositivity at birth of 52.8%, increasing to 97.2% by 12 years of age (p = 0.037). Exposure of cattle to L. interrogans serovar Hardjo was more prevalent in females compared to males, was associated with higher titres of BVDV, and was more prevalent in the wet season compared to the dry season. Exposure of cattle to BVDV was more prevalent in males compared to females, the wet and dry seasons were comparable, and was associated with rising antibody titres against N. caninum and L. interrogans serovar Hardjo. The risk factor survey identified that the probability of herds being N. caninum positive increased with farmer age, if farmers believed there were rodents on farm, and if farmers weren't aware that canids or rodents could contaminate bovid feed on their farm. The probability of a herd being positive to L. interrogans serovar Hardjo increased on farms where multiple cows shared the same bull, where farmers had lower husbandry knowledge, and on farms that used water troughs. The probability of a herd being BVDV seropositive increased with increasing herd size and increasing titres to N. caninum. The benchmarking of bovid exposure to emerging abortifacient pathogens and identification of their risk factors potentially informs disease prevention strategies, supporting efforts to establish a biosecure beef supply for enhanced smallholder livestock productivity, public health and food security in Laos and surrounding countries.

Risk factors for emergence of exotic foot-and-mouth disease O/ME-SA/Ind-2001d on smallholder farms in the Greater Mekong Subregion.

Foot-and-mouth disease (FMD) is a significant endemic transboundary animal disease in Lao People's Democratic Republic (Lao PDR) and throughout the Greater Mekong Subregion (GMS). The disease has been shown to perpetuate the cycle of smallholder poverty through reduced animal production, plus limitations on market access for trading in livestock and their products. Despite significant national and multilateral efforts to control FMD over the past two decades, endemic FMD viruses (FMDVs) continue to circulate in Lao PDR. Further, the threat from new and emerging FMDVs is increasing as transboundary movements in the region intensify in response to increasing regional demand for meat. Although the economic impacts of FMD on smallholder farmers in Lao PDR are significant, studies investigating household-level risk factors for FMD are lacking. Following an outbreak of a novel FMDV (O/ME-SA/Ind2001d) in Lao PDR in 2015, a questionnaire and serological study were conducted in Naxaythong District to identify household-level risk factors associated with this outbreak, as well as endemic circulating viruses in the outbreak area. Data were analysed using a multivariable generalised estimating equation (GEE) model with a logit link function and associations were calculated as odds ratios (OR) with 95% confidence intervals (CI95%). After adjusting for other variables, the practice of quarantining new livestock for a minimum of two weeks prior to introduction to a herd was found to be a significant protective factor during the 2015 outbreak (OR 0.225, CI95% [0.06, 0.88], p-value 0.003). In addition, households owning one or more animals with titres to the non-structural proteins of FMDV, indicating prior infection, had 5.5 times the odds (CI95% [6.16, 49.11], p-value <0.001) of sharing communal grazing land with neighbouring villages. These findings indicate that implementing basic on-farm biosecurity and improved husbandry measures to minimise FMDV circulation at the household level are important and reinforce the need to enhance the education of smallholder farmers in infectious disease control.

Serosurveillance of Coxiellosis (Q-fever) and Brucellosis in goats in selected provinces of Lao People's Democratic Republic.

Goat raising is a growing industry in Lao People's Democratic Republic, with minimal disease investigation to date, especially zoonoses. This study determined the proportional seropositivity of two zoonotic diseases: Q fever (causative agent Coxiella burnetii) and Brucellosis (Brucella species) in goats across five provinces (Vientiane Capital, Xayaboury, Xiengkhuang, Savannakhet and Attapeu). A total of 1458 goat serum samples were tested using commercial indirect ELISA for both pathogens, plus Rose Bengal agglutination test for Brucellosis. Overall individual seropositivity of C. burnetii was 4.1% and Brucella spp. was 1.4%. A multiple logistic regression model identified that province (Vientiane Capital, p = 0.05), breed (introduced Boer mixed breed, p = 0.006) and age (goats ≥3 years old, p = 0.014) were significant risk factors for C. burnetii seropositivity. The results of the survey indicated that province (Vientiane Capital, p<0.001), breed (introduced Boer mixed breed, p<0.001), production system (commercial, p<0.001), age (adult, p = 0.004), and farm size (large, 0.001) were all significant risk factors seropositivity for Brucella spp. It was concluded that Lao goats have been exposed to both C. burnetii and Brucella spp. however the risk of clinical disease has not yet been determined and there is an urgent need to determine human health risks and economic losses caused by Q fever and Brucellosis.

Neuronal ceroid lipofuscinosis in Devon cattle is caused by a single base duplication (c.662dupG) in the bovine CLN5 gene.

The neuronal ceroid lipofuscinoses (NCLs, Batten disease) are recessively inherited neurodegenerative disorders that affect humans and other animals, characterised by brain atrophy and the accumulation of lysosome derived fluorescent storage bodies in neurons and most other cells. Common clinical signs include blindness, ataxia, dementia, seizures and premature death. The associated genes for six different human forms have been identified (CLN1, CLN2, CLN3, CLN5, CLN6 and CLN8), and three other human forms suggested (CLNs 4, 7 and 9). A form of NCL in Australian Devon cattle is caused by a single base duplication (c.662dupG) in bovine CLN5. This mutation causes a frame-shift and premature termination (p.Arg221GlyfsX6) which is predicted to result in a severely truncated protein, analogous to disease causing mutations in human Finnish late infantile variant NCL (CLN5), and a simple genetic diagnostic test has been developed. The symptoms and disease course in cattle also matches CLN5. Only one initiation site was found in the bovine gene, equivalent to the third of four possible initiation sites in the human gene. As cattle are anatomically and physiologically similar to humans with a human-like central nervous system and easy to maintain and breed, they provide a valuable alternative model for CLN5 studies.

Topical anaesthesia reduces sensitivity of castration wounds in neonatal piglets.

The aim of this study was to do determine the efficacy of the topical anaesthetic Tri-Solfen® in the amelioration of the pain of castration in piglets. The trial was conducted over a three day period, and blocked across six litters with 12 piglets treated on days one and two, and 16 on day three. The piglets were randomly allocated by weight and litter to 1 of 4 treatment groups: (i) sham castration (SHAM; n = 10); (ii) surgical castration with no anaesthetic intervention (CAST; n = 10); (iii) surgical castration with post-operative topical anaesthesia (TRI; n = 10); (iv) surgical castration with a pre-operative intra-testicular lignocaine hydrochloride injection (LIG; n = 10). Wound sensitivity testing involved von Frey monofilaments of weights 4g and 300g, and an 18 gauge needle, used to stimulate the wound and surrounding skin respectively, at various pre-determined sites. Neonatal piglets receiving topical anaesthesia (Tri-Solfen®) spray into castration wounds had significantly lower wound sensitivity responses for up to 4h, compared to those castrated following intra-testicular lignocaine injection or those with no treatment. The use of topical anaesthetic is suggested as a practical and affordable method of improving piglet welfare during castration.

Changes in prevalence of ovine paratuberculosis following vaccination with Gudair®: Results of a longitudinal study conducted over a decade.

Since its registration in 2002, Gudair® vaccine has become the key tool for managing paratuberculosis in sheep in Australia. This study was conducted to evaluate its effectiveness in reducing paratuberculosis prevalence in 12 flocks over more than one decade. Flocks with variable initial prevalence (5, 4 and 3 with low, medium, and high prevalence, respectively) were enrolled in the study in 2003-04. Six biennial faecal samplings were conducted in these flocks over a 10year period. At each sampling, faeces from four age groups (3, 4, 5 and 6year-old sheep) were collected from each flock. The aim was to select 7 or 14 pools of faeces constituted with pellets from 50 or 25 sheep from each age group, respectively, but the numbers and sizes of pools did vary due to logistical issues. Samples were cultured using pooled faecal culture approach and sheep level paratuberculosis prevalence was estimated. Changes in probability of a pool being positive and in paratuberculosis prevalence over time were evaluated by fitting generalised-linear and linear mixed models. The proportion of positive faecal pools significantly declined over time from 50.3% at the first sampling in 2003-04 to only 3.1% at the last sampling in 2013-14, suggestive of a 30-fold reduction in the odds of a pooled faecal sample to be positive (p<0.001). Similarly, the average animal level prevalence in flocks dropped from 7.64% at the first sampling to 0.12% at the last sampling. However, faecal shedding persisted and was still present in 3 of the 8 flocks (37.5%) that remained in the study at the last sampling conducted in 2013-14. This is the longest study ever conducted to evaluate paratuberculosis vaccine effectiveness. The results will enable sheep producers to make informed decisions on managing paratuberculosis, and in evaluating the risks of purchasing and trading vaccinated sheep.

Virus-induced congenital malformations in cattle.

Diagnosing the cause of bovine congenital malformations (BCMs) is challenging for bovine veterinary practitioners and laboratory diagnosticians as many known as well as a large number of not-yet reported syndromes exist. Foetal infection with certain viruses, including bovine virus diarrhea virus (BVDV), Schmallenberg virus (SBV), blue tongue virus (BTV), Akabane virus (AKAV), or Aino virus (AV), is associated with a range of congenital malformations. It is tempting for veterinary practitioners to diagnose such infections based only on the morphology of the defective offspring. However, diagnosing a virus as a cause of BCMs usually requires laboratory examination and even in such cases, interpretation of findings may be challenging due to lack of experience regarding genetic defects causing similar lesions, even in cases where virus or congenital antibodies are present. Intrauterine infection of the foetus during the susceptible periods of development, i.e. around gestation days 60-180, by BVDV, SBV, BTV, AKAV and AV may cause malformations in the central nervous system, especially in the brain. Brain lesions typically consist of hydranencephaly, porencephaly, hydrocephalus and cerebellar hypoplasia, which in case of SBV, AKAV and AV infections may be associated by malformation of the axial and appendicular skeleton, e.g. arthrogryposis multiplex congenita. Doming of the calvarium is present in some, but not all, cases. None of these lesions are pathognomonic so diagnosing a viral cause based on gross lesions is uncertain. Several genetic defects share morphology with virus induced congenital malformations, so expert advice should be sought when BCMs are encountered.

Identification of a plant isoflavonoid that causes biliary atresia.

Biliary atresia (BA) is a rapidly progressive and destructive fibrotic disorder of unknown etiology affecting the extrahepatic biliary tree of neonates. Epidemiological studies suggest that an environmental factor, such as a virus or toxin, is the cause of the disease, although none have been definitively established. Several naturally occurring outbreaks of BA in Australian livestock have been associated with the ingestion of unusual plants by pregnant animals during drought conditions. We used a biliary secretion assay in zebrafish to isolate a previously undescribed isoflavonoid, biliatresone, from Dysphania species implicated in a recent BA outbreak. This compound caused selective destruction of the extrahepatic, but not intrahepatic, biliary system of larval zebrafish. A mutation that enhanced biliatresone toxicity mapped to a region of the zebrafish genome that has conserved synteny with an established human BA susceptibility locus. The toxin also caused loss of cilia in neonatal mouse extrahepatic cholangiocytes in culture and disrupted cell polarity and monolayer integrity in cholangiocyte spheroids. Together, these findings provide direct evidence that BA could be initiated by perinatal exposure to an environmental toxin.