Novel estimation of African swine fever transmission parameters within smallholder villages in Lao P.D.R.

African Swine Fever (ASF) disease transmission parameters are crucial for making response and control decisions when faced with an outbreak, yet they are poorly quantified for smallholder and village contexts within Southeast Asia. Whilst disease-specific factors - such as latent and infectious periods - should remain reasonably consistent, host, environmental and management factors are likely to affect the rate of disease spread. These differences are investigated using Approximate Bayesian Computation with Sequential Monte-Carlo methods to provide disease parameter estimates in four naïve pig populations in villages of Lao People's Democratic Republic. The villages represent smallholder pig farmers of the Northern province of Oudomxay and the Southern province of Savannakhet, and the model utilised field mortality data to validate the transmission parameter estimates over the course of multiple model generations. The basic reproductive number between-pigs was estimated to range from 3.08 to 7.80, whilst the latent and infectious periods were consistent with those published in the literature for similar genotypes in the region (4.72 to 6.19 days and 2.63 to 5.50 days, respectively). These findings demonstrate that smallholder village pigs interact similarly to commercial pigs, however the spread of disease may occur slightly slower than in commercial study groups. Furthermore, the findings demonstrated that despite diversity across the study groups, the disease behaved in a consistent manner. This data can be used in disease control programs or for future modelling of ASF in smallholder contexts.

Effect of Spirulina platensis Supplementation on Reproductive Parameters of Sahrawi and Jabbali Goat Bucks.

Spirulina platensis (SP) is a protein-rich dietary supplement that improves animal reproductive traits. This study investigated the effect of SP supplementation on puberty onset, semen characteristics, scrotal circumference (SC), libido, and hormone concentrations in Sahrawi and Jabbali bucks. The study was conducted in 36 bucks, divided into three groups (n = 6/group), for 70 days. The rations included the following: (1) Control feed (Con) with 14% crude protein and 11.97% MJ/kg DM energy; (2) Con with 2 g SP/head/day SP treatment (T1) and (3) Con with 4 g SP/head/day treatment (T2). The mean (±SEM) SC of both SP groups in the Sahrawi breed was significantly higher (p ≤ 0.05) compared to the Con. The mean of the semen volume significantly increased (p ≤ 0.05) in the SP group than in the Con group in both breeds. SP groups vs. Con groups had increased sperm concentration in Sahrawi bucks than Jabbali bucks. Mean serum luteinizing hormone (LH) and testosterone (Tes) concentrations in Jabbali bucks were significantly higher (p ≤ 0.05) in the SP groups compared to Sahrawi bucks. SP improved the SC, semen quality, libido, sperm concentration, and LH and Tes concentrations in both breeds. The results of the current study suggest that adding SP to the diet may have the ability to improve the semen quality of the local Omani bucks.

Retrospective investigation of the 2019 African swine fever epidemic within smallholder pig farms in Oudomxay province, Lao PDR.

The 2019 African swine fever (ASF) outbreak in the Lao People's Democratic Republic (Lao PDR or Laos) represented a major epidemiologic event where a transitioning lower-middle income nation (LMIC) experienced a viral epidemic in a naïve pig population. The diversity of pig management styles creates challenges for local and regional policymakers when formulating recommendations to control an ASF outbreak. The aim of this study were to investigate the management of pigs in villages of Oudomxay province that were affected by ASF in 2019, as a case study in a smallholder pig-rasing system in northern Laos. The frequencies of well known risk factors were measured in the affected villages and the timelines and household level stock losses due to the outbreak were investigated. These findings were compared to data available from a similar outbreak in the southern province of Savannakhet. Disease control implications of these findings are discussed. Mean losses were 3.0-23.3 pigs per household, with a mean lost herd value of USD 349, 95% CI (294-415). These pig losses reflect those estimated in Savannakhet (6.7 pigs per household). However, the financial loss estimated per household was higher, USD 349 versus USD 215, possibly due to higher pig values and a higher input/output management approach in Oudomxay. The investigation revealed the presence of numerous ASF risk factors, such as swill-feeding and free-ranging. In addition, poor biosecurity practices - such as inappropriate garbage disposal and slaughtering - that could contaminate the environment were present. ASF cases occurred across all villages between June and December 2019, with outbreak periods ranging from 22-103 days. These values are consistent with the outbreak in Savannakhet; however, notable differences in management styles were observed. These findings demonstrate the need for more disease control resources from the village to the Governmental level. Villages need support in enacting context appropriate biosecurity measures, whilst the ongoing surveillance and investigation of ASF require investment in logistical and veterinary resources at the Governmental level.

Effect of Spirulina platensis Supplementation on Carcass Characteristics, Fatty Acid Profile, and Meat Quality of Omani Goats.

In a 70-day study, 36 Jabbali and Sahrawi bucks, aged 11 months, were utilized to evaluate the effects of different levels of spirulina dietary supplement (SP) on carcass characteristics, fatty acid profile, and meat quality traits in Omani goat breeds. The goats were put into six groups of six bucks, each at random. The diet consisted of a conventional concentrate feed ration (CFR) without spirulina (CON), and the CFR diet supplemented with spirulina at the levels of 2 g/head daily (T1) and 4 g/head daily (T2). In general, Sahrawi bucks showed a highly significant response to SP feeding compared with Jabbali bucks. The treatment groups, especially T1, showed a significant increase in average daily gain and carcass traits (body length, leg length, and the rack weight) compared with the CON group of Sahrawi bucks. The weights of omental and kidney fat were also significantly higher in T1 compared with CON and T2 groups of Sahrawi goats, while they were significantly higher in T2 compared with CON and T1 groups of Jabbali goats. Carcass profile and meat quality, including ultimate ph and meat color lightness (L*) were increased significantly with dietary spirulina in both LD and SM muscles of Sahrawi goats. Most of the Sfa, Mufa, Pufa, Pufa n-6, Pufa n-3, and n-6/n-3 ratios of the LD showed significant differences in diets supplemented with SP compared with CON for Sahrawi bucks, while some of them were significant in Jabbali bucks. The LD muscle of Sahrawi goats fed diets supplemented with SP of the T1 group significantly decreased in the amounts of pentadecanoic and margaric acids compared with the T2 and CON groups. The study concluded that incorporating SP (2 g and 4 g/head daily) into the diet of Omani goats, especially Sahrawi goats, can increase growth performance, as well as improve fatty acid composition and meat quality.

Utilising abattoir sero-surveillance for high-impact and zoonotic pig diseases in Lao PDR.

National disease surveillance systems are essential to a healthy pig industry but can be costly and logistically complex. In 2019, Lao People's Democratic Republic (Lao PDR) piloted an abattoir disease surveillance system to assess for the presence of high impact pig diseases (HIPDs) using serological methods. The Lao Department of Livestock and Fisheries (DLF) identified Classical Swine Fever (CSF), Porcine Respiratory and Reproductive Syndrome (PRRS) and Brucella suis as HIPDs of interest for sero-surveillance purposes. Porcine serum samples (n = 597) were collected from six Lao abattoirs in March to December of 2019. Serological enzyme-linked immunosorbent assay (ELISA) methods were chosen for their high-throughput and relatively low-costs. The true seroprevalence for CSF and PRRS seropositivity were 68.7%, 95% CI (64.8-72.3) and 39.5%, 95% CI (35.7-43.5), respectively. The results demonstrated no evidence of Brucella spp. seroconversion. Lao breed pigs were less likely to be CSF seropositive (P < 0.05), whilst pigs slaughtered at <1 year of age were less likely to be PRRS seropositive (P < 0.01). The testing methods could not differentiate between seropositivity gained from vaccine or natural infection, and investigators were unable to obtain the vaccine status of the slaughtered pigs from the abattoirs. These results demonstrate that adequate sample sizes are possible from abattoir sero-surveillance and lifetime health traceability is necessary to understand HIPDs in Lao PDR.

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.

Foot-and-mouth disease seroprevalence and reporting behaviours in nine northern provinces in Lao PDR: The current situation and challenges for control.

Foot-and-mouth disease (FMD), caused by the FMD virus (FMDV), is one of the most important of global livestock diseases, impacting even-toed ungulates and distributed mostly in less developed countries that are home to 75% of the human population. A progressive control pathway for FMD (PCP-FMD) was developed to assist countries to better manage FMD risks and progress towards control and eradication. This requires evidence of current FMD seroprevalence to enable the informed risk assessment and the disease control planning required to progress along the initial stages of the PCP-FMD. Despite Laos being an active member of the South East Asia and China FMD campaign, these data have been challenging to obtain. To address this gap, a cross-sectional seroprevalence study in nine northern Lao provinces was conducted in early 2019. The study sampled 602 large ruminants and 19 goats from 30 villages. Overall, the large ruminant seroprevalence was 43.7% (95% CI 39.7-47.8), and 90% of sampled villages had at least one seropositive animal. Large ruminants suspected of having had clinical FMD in their lifetime were found to have an increased risk of being seropositive (odds ratio 1.96, 95% CI 1.06-3.65, p = .03). An examination of current reporting behaviours of farmers, village veterinary workers and district and provincial officials found that individuals were using their discretion as to whether disease events required reporting along the chain of command. This, plus the lack of a computerized reporting system, contributed to a loss of historical data, potentially compromising progression on the PCP-FMD. Laos requires ongoing support from donor agencies to improve the current animal disease surveillance system and implement effective FMD control strategies that can enable progress on the PCP-FMD.

Changes in farmer animal health and biosecurity knowledge, attitudes and practices: Insights from Cambodia and Laos.

Livestock agriculture in Cambodia and Laos is severely affected by endemic and exotic transboundary animal diseases, impacting household livelihoods and food and nutritional security. Collaborative animal health and biosecurity projects were conducted in each country between 2015 and 2019 aimed at increasing smallholder livestock production through mainly knowledge-based interventions in large ruminant disease prevention, nutrition, reproduction and marketing access. This study's objectives were to identify baseline animal health and biosecurity knowledge, attitudes and practices (KAP) of farmers, and temporal changes in key attitudes and practices associated with improved knowledge. Data for each country were analysed separately, and a formal between-country comparison was avoided due to major implementation and management differences. Binomial logistic regression without random terms compared the overall knowledge scores between 2015 and 2018 in each country, and binomial logistic regression with random terms was used to investigate possible associations between animal health and biosecurity knowledge scores and attitudes and practices. In Cambodia, the average animal health and biosecurity knowledge score increased from 4.8 (±2.2)/17 in 2015 to 13.0 (±2.8)/17 in 2018 (p < .0001). In Laos, the average scores improved from 7.2 (±2.5)/12 in 2015 to 7.9 (±2.9)/12 in 2018 (p = .0003). Clinical signs of foot-and-mouth disease were accurately reported by 100% of Cambodian and 88% of Lao project farmers, and overall, attitudes and practices improved in both countries over the study period. The belief that vaccination can improve livestock value was associated with increasing knowledge in both countries (Cambodia: OR 1.45, 95% CI 1.24-1.68, p < .0001; Laos: OR 1.43, 95% CI 1.16-1.75, p = .0006) as was the use of the services of village para-veterinarians if they had better training (Cambodia: OR 1.76, 95% CI 1.20-2.58, p = .004; Laos: OR 2.02, 95% CI 1.19-3.41, p = .009). However, the source of livestock information (District veterinary officials) was the only practice associated with knowledge in both Cambodia and Laos. The influence of resource scarcity on farmer decision making and knowledge measures was considered of particular relevance to the findings.

Thirteen Cases of Valeryl Fentanyl in Michigan: A Call for Expanding Opioid Testing.

ABSTRACT: In this report, we describe 13 cases of drug overdose in Michigan in which valeryl fentanyl was found in postmortem blood. Valeryl fentanyl is a schedule I opioid that is rarely found in drug overdoses in the United States. Although little data exist on the mortality and morbidity associated with valeryl fentanyl, its molecular structure indicates that it would be less potent than fentanyl.When analyzing blood samples for valeryl fentanyl, samples from peripheral sites were sometimes negative for quantitative levels; however, samples from central sites in the same decedent were positive. This could indicate unique pharmacokinetics for valeryl fentanyl, which could have implications for other fentanyl analogs. Given the paucity of pharmacodynamic information, the prohibition of its use, the potential to buttress law enforcement efforts in monitoring drug trafficking trends, and to determine the efficacy of current regulations, laboratories should test for valeryl fentanyl. When testing for valeryl fentanyl, and likely other fentanyl analogs, the site of sample collection is important: central sources of blood are preferred to peripheral sources.

Are infectious reproductive pathogens of large ruminants a threat to improving food security? An investigation from Cambodia.

In Cambodia, information on common pathogens causing reproductive losses in cattle and buffalo are lacking, despite there being a need to address livestock health to enhance food security. We analysed stored buffalo (n = 29) and cattle (n = 471) serum samples collected in 2016 using commercially available enzyme-linked immunosorbent assay kits. Antibodies to Neospora caninum, bovine viral diarrhoea virus (BVDV), Leptospira interrogans serovar Hardjo and Brucella abortus were detected in buffalo samples at 79.3% (95% CI 64.6-94.0), 3.4% (95% CI 0-10.0), 0% and 0%, respectively, and in cattle at 4.2% (95% CI 2.4-6.0), 6.4% (95% CI 4.2-8.6), 8.1% (95% CI 5.6-10.6) and 0%, respectively. The high N. caninum seroprevalence in buffalo was associated with increasing age, with buffalo having a 13.1% chance of being seropositive at birth, increasing to 99.4% by age 7 (p = 0.045). This suggests a predominance of horizontal transmission, possibly from exposure to faeces from free-roaming village dogs. Cattle L. interrogans serovar Hardjo seroprevalence was highest in Tbong Khmum province (20.4%) compared to other provinces (p < 0.001), and may be compromising bovine fertility and creating a zoonotic risk for smallholders who may contract leptospirosis from farm work. These high infection rates prompt further research to determine: to what extent these pathogens are linked to the low reproductive efficiency observed in large ruminants in Cambodia, the risk factors to pathogen exposure and appropriate strategies to reduce these risks. Low BVDV and B. abortus exposure is an important observation. Increasing large ruminant livestock trade into the country will require improved biosecurity and disease surveillance to prevent their emergence. An enhanced understanding of the status of infectious reproductive livestock pathogens in Cambodia can assist development projects to make evidence-based strategies to enhance cattle and buffalo health and improve food security.

The impact of African swine fever virus on smallholder village pig production: An outbreak investigation in Lao PDR.

African swine fever virus (ASFV) causes a deadly disease of pigs which spread through southeast Asia in 2019. We investigated one of the first outbreaks of ASFV in Lao People's Democratic Republic amongst smallholder villages of Thapangtong District, Savannakhet Province. In this study, two ASFV affected villages were compared to two unaffected villages. Evidence of ASFV-like clinical signs appeared in pig herds as early as May 2019, with median epidemic days on 1 and 18 June in the two villages, respectively. Using participatory epidemiology mapping techniques, we found statistically significant spatial clustering in both outbreaks (p < 0.001). Villagers reported known risk factors for ASFV transmission - such as free-ranging management systems and wild boar access - in all four villages. The villagers reported increased pig trader activity from Vietnam before the outbreaks; however, the survey did not determine a single outbreak source. The outbreak caused substantial household financial losses with an average of nine pigs lost to the disease, and Monte Carlo analysis estimated this to be USD 215 per household. ASFV poses a significant threat to food and financial security in smallholder communities such as Thapangtong, where 40.6% of the district's population are affected by poverty. This study shows ASFV management in the region will require increased local government resources, knowledge of informal trader activity and wild boar monitoring alongside education and support to address intra-village risk factors such as free-ranging, incorrect waste disposal and swill feeding.

Meat of South American camelids - Sensory quality and nutritional composition.

Meat contains a range of nutrients in a highly bioavailable form and when meat is excluded from the diet, without being replaced with suitable alternatives, nutrient deficiencies may occur. For this reason, it is very important to extend our knowledge of nutrients in alternative red meats, such as that provided from the South American camelids- llama and alpaca. This review summarises the current information on the nutritional and sensory parameters of llama and alpaca meat and factors affecting quality. South American camelids produce lean carcases, with an uneven fat distribution across the carcase. Llama and alpaca meat quality traits are mostly influenced by animal nutrition, animal age and processing methods. A feeding strategy based on pasture and hay supplement of barley and alfalfa, and processing younger animals (18 months) has improved the fatty acid composition respectively in llama and alpaca meat, whereas meat colour and tenderness are influenced by processing treatments including electrical stimulation, tenderstretching and enzymatic infusion.

Managing Welfare and Antimicrobial-Resistance Issues in Treating Foot-and-Mouth Disease Lesions: A New Therapeutic Approach.

INTRODUCTION: Foot-and-mouth disease (FMD) causes negative impacts on global food security, the livestock trade, national economies, and farming-family livelihoods, particularly in resource-poor developing countries with inadequate biosecurity and low levels of vaccination from inadequate veterinary services. As smallholder farmers have limited understanding of disease-risk management, their focus in FMD outbreaks is on accessing clinically effective therapies. However, most are provided inappropriate traditional treatments and/or topical or parenteral antibiotics, often delivered by paraveterinarians inadequately trained in antimicrobial custodianship. This results in negative financial impacts on livelihoods plus risks of food safety and development of antimicrobial resistance. We report the use of a novel pain-relief therapy for FMD. METHODS: Clinical examinations in an outbreak of suspected FMD in April 2019 in Muang Khay village in Luang Prabang province, Laos confirmed signs and lesions of severe, subacute, ulcerative glossitis and interdigital dermatitis, typical of FMD. All affected buffalo (n=99) and cattle (n=37) presented for treatment in a population of 238 large ruminants, from 15 of 136 households, were administered a topical anesthetic pain-relief product (PRP) wound gel by spray-on (10-30 mL per animal) formulation developed for aversive husbandry procedures (Tri-Solfen, Animal Ethics, Australia). RESULTS: Treatment with PRP resulted in immediate improvement in demeanor and locomotion, and no adverse events were observed. On follow-up interview, all owners confirmed that their animals were eating within 2 days and lesions had healed within 5 days. Having experienced the positive clinical impacts of PRP on affected animals, these and surrounding farmers were keen to purchase the PRP for future use. The veterinary authorities rapidly registered the PRP for FMD therapy in Laos due to the observed efficacy. DISCUSSION: These findings suggest a potential paradigm shift from treating FMD with expensive antimicrobials, which risks antimicrobial resistance, to a new, less expensive therapeutic approach that reduces animal suffering and may motivate farmers to report disease to access treatment. Use of the PRP is suggested as an innovation that may improve future FMD management, particularly in developing countries.

Serological Evidence of Foot-and-Mouth Disease Infection in Goats in Lao PDR.

Foot and Mouth Disease (FMD) causes significant economic loss in Lao PDR (Laos) and perpetuates the cycle of smallholder poverty mainly through large ruminant productivity losses, increased costs of production and potential limitations to market access for trade in livestock and their products. Goats are emerging as an important livestock species in Laos, and there is an increasing trend in the number of households with goats, often farmed alongside cattle and buffalo. Although an FMD susceptible species, very little is known about the role of goats in the epidemiology of the disease in Laos. A cross-sectional seroprevalence study was conducted by detecting antibodies to the non-structural proteins (NSP), an indication of a previous infection, and serotype-specific structural proteins (SP) that could be due to vaccination or infection. The study commenced in late 2017 and sera were collected from 591 goats in 26 villages of northern, central and southern Laos. For a subset of sera samples, paired oral swab samples were also collected by a simple random sampling method to detect the prevalence of FMD virus infection at the time of collection. The NSP seroprevalence in the provinces of Borkeo and Xayabouli in the north was 42 and 8%, respectively and in Khammoune in the center, it was 20%. In the other five provinces, Luang Namtha and Luang Prabang (northern Laos), Xieng Khouang and Savannaket (central Laos), and Champasak (southern Laos), the seroprevalence was close to zero. The multivariable analysis indicated that age (p < 0.001) was positively associated with animal-level seropositivity and males were less likely to be seropositive than females (OR: 0.29; 95%CI: 0.10-0.83; p = 0.017). Continued sero-surveillance for FMD in goats is recommended to improve our understanding of their role in the epidemiology of FMD in the region and to extend support to FMD control decisions, particularly regarding vaccination.

Evaluation of the diagnostic accuracy of an affordable rapid diagnostic test for African Swine Fever antigen detection in Lao People's Democratic Republic.

African Swine Fever (ASF) is a transboundary animal disease of pigs and wild suids that appeared in Lao People's Democratic Republic (Lao PDR) in mid-2019, having spread across China and Vietnam in the months prior. Despite the scale of the Asian ASF pandemic and the availability of pen-side rapid diagnostic tests (RDT) on the market, few locally produced and easily available ASF RDTs have been evaluated for diagnostic accuracy. In this study, an ASF antigen detection RDT from Shenzhen Lvshiyuan Biotechnology Co. Ltd was evaluated using clinical field samples submitted to the National Animal Health Laboratory (NAHL) from ASF suspect cases between June and December 2019 in Lao PDR. Positive (n = 57) and negative (n = 50) samples of whole blood, serum and haemolysed serum were assessed by RDT and PCR, with the latter used as the gold standard reference comparator. Overall the RDT had a diagnostic sensitivity (DSe) of 65 %, 95 % CI [51-77] and diagnostic specificity (DSp) of 76 %, 95 % CI [62-87]. The RDT demonstrated improved performance on samples with lower PCR cycle threshold (ct) values with each additional cycle reducing the odds of the RDT returning a positive by 17 % relative to the previous cycle, 95 % CI [8 %-28 %] (P < 0.01). While this test shows promise for field application, complete validation of diagnostic accuracy requires a larger sample size.

The effect of whole carcase medium voltage electrical stimulation, tenderstretching and longissimus infusion with actinidin on alpaca meat quality.

The effect on alpaca meat quality from applying medium voltage electrical stimulation (ES) in combination with tenderstretching (TS; pubic symphysis suspended) to whole carcases was investigated, along with the effect of actinidin infusion on alpaca longissimus (LTL) quality. Carcases (n = 36) were allocated to either no ES + achilles hung; or ES + TS. The left- and right-hand side LTL of each carcase was allocated to one of three infusion treatments; no infusion (control), infusion with water or infusion with enzyme. Processing treatments reduced LTL and semimembranosus shear force without negatively impacting colour or oxidation traits. Infusion with enzyme reduced LTL shear force relative to control and water treatments but resulted in reduced consumer acceptance. The use of TS with ES in commercial alpaca processing is supported. There was no advantage to infusing alpaca LTL with actinidin as results indicate a net negative effect on consumer acceptance of this novel meat.

Which species is in the faeces at a time of global livestock movements: single nucleotide polymorphism genotyping assays for the differentiation of Fasciola spp.

Fasciolosis, caused by Fasciola hepatica and Fasciola gigantica, is a globally distributed zoonotic disease of livestock. While F. hepatica and F. gigantica have temperate and tropical distributions, respectively, parasite sympatry occurs in parts of Asia and Africa. A growing protein demand has the potential to facilitate the translocation of parasites from endemic to non-endemic areas, via associated international livestock movements. Such is the case in Southeast Asia, where livestock trade from F. hepatica-endemic countries into China and Vietnam may account for detection of F. hepatica hybrid/introgressed forms. Of particular importance is Lao People's Democratic Republic, which acts as a major livestock thoroughfare for the region. Our ability to understand the impacts of livestock-associated Fasciola spp. movements on local animal and human health is hindered by a lack of ante-mortem diagnostic tools allowing species differentiation. Molecular tools have been developed for Fasciola spp. differentiation, however those rely on access to pure DNA from adult specimens, limiting their application to post-mortem use. Our aim was to detect and differentiate F. hepatica from the endemic F. gigantica in local smallholder cattle in a region of Southeast Asia with frequent livestock trafficking. To do this we designed and validated ante-mortem molecular assays for Fasciola spp. differentiation targeting single-nucleotide polymorphisms (SNPs) within ITS1 and lsrRNA. We then deployed these SNP genotyping assays to diagnose Fasciola spp. infection in 153 local cattle from 27 villages in Northern Laos. We demonstrate the presence of F. hepatica DNA, confirmed by qualitative Sanger and quantitative Illumina amplicon sequencing of ITS1 and lsrRNA, and highlight the shortfalls of Sanger sequencing for Fasciola spp. identification due to the preferential amplification of F. gigantica nucleotides in mixed DNA samples. The outlined protocol enables rapid surveillance of faecal samples for the presence of Fasciola species eggs, their co-infection and/or infection with F. hepatica/F. gigantica hybrids.

The effects of season and post-transport rest on alpaca (Vicunga pacos) meat quality.

The effects of season (summer, autumn, winter and spring) and post-transportation rest on alpaca meat quality were investigated in 160 castrated male alpacas (23 ±â€¯1 month of age) over a 12 month period. Twice per season, animals were randomly allocated to consignment groups of 20, transported 4 h to slaughter and allocated to either of two treatment groups: (1) overnight lairage pre slaughter (Direct) and (2) seven day rest period with access to feed pre slaughter (Rested). At slaughter, blood was collected for the analysis of plasma cortisol and a longissimus core sample obtain pre rigor for glycogen content determination. Alpaca muscle moisture loss increased through summer and spring in the longissimus thoracics. Seasonal differences did not reflect pasture seasonality or muscle glycogen content. Resting alpacas for 7 days pre-slaughter reduced muscle glycogen content and tenderness. Drip loss and purge was greater for rested animals indicating that resting alpacas post transport is not advantageous to alpaca meat quality.

Improving Village Animal Health Worker participation in national disease surveillance systems: A case study from Cambodia.

Para-veterinary systems have arisen in numerous developing countries to address the low capacity of national veterinary services in meeting livestock health demands of mainly smallholder farmers. In Cambodia, the village animal health worker (VAHW) system was established in the early 1990s, involving short training programmes to equip VAHWs to provide basic animal health services for smallholder farmers, particularly the vaccination of cattle for haemorrhagic septicaemia (HS). However, there are increasing expectations that VAHWs provide village level disease surveillance information to the national veterinary services, despite their low-level disease diagnostic skills. To identify opportunities to improve the disease reporting system in Cambodia, a closed-ended cross-sectional study of VAHWs (n = 80) from two provinces was conducted in 2015, examining their contact frequency with district and provincial animal health authorities. Ordinal logistic regression was used to determine factors associated with increased frequency of VAHW contact with the district animal health authorities responsible for national disease reporting. Positive associations between income generated from VAHW activities (p = .01) and the frequency of visiting farmers (p < .0001), with more frequent contacts with district animal health authorities, were identified. High levels of inappropriate use of antibiotics, with almost 90% of VAHW reporting they use antibiotics to treat FMD-affected animals, were reported. Although further research to increase the rates of reporting at the smallholder farmer and VAHW levels is required, increased emphasis on complying with the World Organisation for Animal Health's competency guidelines for veterinary paraprofessionals may be required if they are expected to contribute to national animal disease surveillance systems.

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.