Stress steroid levels and the short-term impact of routine dehorning in female southern white rhinoceroses (Ceratotherium simum simum)

Rhinoceros populations in Africa are under severe threat as a result of surging poaching rates and risk-mitigation strategies are continuously adapted in an attempt to ensure the survival of the species. This study compared faecal glucocorticoid metabolite (fGCM) levels of two age classes of limited free-ranging female white rhinos with fGCM levels of adult free-ranging female white rhinos. Subsequently, fGCM alterations in the limited free-ranging animals were monitored following routine dehorning as a measure of the animals’ short-term physiological stress response. Baseline fGCM levels differed significantly between tested groups, with both free-ranging and limited free-ranging adult animals showing significantly higher fGCM levels compared with limited free-ranging juvenile females. In contrast, baseline fGCM levels did not differ significantly between limited free-ranging and free-ranging adult individuals. Routine dehorning procedures resulted in a short-term stress response expressed by a significant increase in fGCM levels 48 h post-dehorning, with stress steroid levels returning to pre-dehorning concentrations 72 h after the procedure.http://www.nisc.co.za/products/59/journals/african-zoology2017-12-31hb2017Anatomy and Physiolog

Detection of equine herpesvirus-4 and physiological stress patterns in young Thoroughbreds consigned to a South African auction sale

BACKGROUND : The prevalence of equine herpesvirus types-1 and -4 (EHV-1 and -4) in South African Thoroughbreds at auction sales is currently undefined. Commingling of young Thoroughbreds from various populations together with physiological stress related to their transport and confinement at a sales complex, may be associated with shedding and transmission of EHV-1 and -4. This prospective cohort study sampled 90 young Thoroughbreds consigned from eight farms, originating from three provinces representative of the South African Thoroughbred breeding demographic to a sales complex. Nasal swabs for quantitative real-time polymerase chain reaction (qPCR) assay to detect EHV-1 and -4 nucleic acid and blood samples for enzyme-linked immunosorbent assay for EHV-1 and -4 antibodies were collected from all horses on arrival and departure. Additional nasal swabs for qPCR were obtained serially from those displaying pyrexia and, or nasal discharge. Daily faecal samples were used for determination of faecal glucocorticoid metabolite (FGM) concentrations as a measurement of physiological stress and these values were modelled to determine the factors best explaining FGM variability. RESULTS : EHV-4 nucleic acid was detected in 14.4 % and EHV-1 from none of the animals in the study population. Most (93.3 %) and very few (1.1 %) of this population showed antibodies indicating prior exposure to EHV-4 and EHV-1 respectively. Pyrexia and nasal discharge were poor predictors for detecting EHV-4 nucleic acid. The horses’ FGM concentrations increased following arrival before decreasing for most of the remaining study period including the auction process. Model averaging showed that variation in FGM concentrations was best explained by days post-arrival and transport duration. CONCLUSIONS : In this study population, sales consignment was associated with limited detection of EHV-4 nucleic acid in nasal secretions, with most showing prior exposure to EHV-4 and very few to EHV-1. The physiological stress response shown by most reflected the combination of stressors associated with transport and arrival and these are key areas for future investigation into management practices to enhance health and welfare of young Thoroughbreds during sales consignment.Racing South Africa (Pty) Ltd, the Equine Research Centre and Departments of Companion Animal Clinical Studies and Production Animal Clinical Studies, Faculty of Veterinary Science, University of Pretoria, South Africa.http://www.biomedcentral.com/bmcvetresam201

First detection and frequent occurrence of Equine Hepacivirus in horses on the African continent

Since the discovery of equine hepacivirus (EqHV) in 2011, the virus has been detected in horse populations from more than twelve countries across five continents. EqHV seroprevalence has been reported to be as high as 61.8% and EqHV ribonucleic acid (RNA) prevalence to range between 0.9% and 34.1%. Molecular and serological indications of EqHV infection have never been reported in equids on the African continent. Therefore, investigation of EqHV prevalence in South African horses and subsequent viral genetic characterization contribute to a better understanding of the global epidemiology of this virus. In a cross-sectional study, serum samples from 454 Thoroughbred foals (aged 58–183 days) were analysed for anti-EqHV non-structural protein 3 (NS3)-specific antibodies (abs) with a luciferase immunoprecipitation system (LIPS) and for EqHV RNA by quantitative real-time polymerase chain reaction (qRT-PCR). Farms of origin (n = 26) were situated in South Africa’s Western Cape Province. The associations between EqHV infection state and farm of origin, foal gender and foal age were subsequently described. Furthermore, nested PCRs were performed on parts of the 5′UTR, NS3 and NS5B genes of 17 samples. Samples were sequenced and phylogenetic analyses were conducted. The population’s seroprevalence was 83.70% and RNA was detected in 7.93% of samples. Increasing foal age was associated with decreasing ab prevalence and increasing prevalence of EqHV RNA. Sequences from South African EqHV strains did not show in-depth clustering with published sequences of EqHV isolates from particular continents. In conclusion, EqHV is present in the South African Thoroughbred population and appears more prevalent than reported in other horse populations worldwide.M.B. was supported by the Deutscher Akademischer Austauschdienst (DAAD). B.T. was supported by the Hannover Medical School. E.S. was supported by the Helmholtz Centre for Infection Research.http://www.elsevier.com/locate/vetmic2019-09-01hj2018Companion Animal Clinical StudiesEquine Research Centr

An Equine Model for Vaccination against a Hepacivirus: Insights into Host Responses to E2 Recombinant Protein Vaccination and Subsequent Equine Hepacivirus Inoculation

Equine hepacivirus (EqHV) is the closest known genetic homologue of hepatitis C virus. An effective prophylactic vaccine is currently not available for either of these hepaciviruses. The equine as potential surrogate model for hepacivirus vaccine studies was investigated, while equine host responses following vaccination with EqHV E2 recombinant protein and subsequent EqHV inoculation were elucidated. Four ponies received prime and booster vaccinations (recombinant protein, adjuvant) four weeks apart (day −55 and −27). Two control ponies received adjuvant only. Ponies were inoculated with EqHV RNA-positive plasma on day 0. Blood samples and liver biopsies were collected over 26 weeks (day −70 to +112). Serum analyses included detection of EqHV RNA, isotypes of E2-specific immunoglobulin G (IgG), nonstructural protein 3-specific IgG, haematology, serum biochemistry, and metabolomics. Liver tissue analyses included EqHV RNA detection, RNA sequencing, histopathology, immunohistochemistry, and fluorescent in situ hybridization. Al-though vaccination did not result in complete protective immunity against experimental EqHV inoculation, the majority of vaccinated ponies cleared the serum EqHV RNA earlier than the control ponies. The majority of vaccinated ponies appeared to recover from the EqHV-associated liver insult earlier than the control ponies. The equine model shows promise as a surrogate model for future hepacivirus vaccine research

Detection of equine herpesvirus-4 and physiological stress patterns in young Thoroughbreds consigned to a South African auction sale

BACKGROUND : The prevalence of equine herpesvirus types-1 and -4 (EHV-1 and -4) in South African Thoroughbreds at auction sales is currently undefined. Commingling of young Thoroughbreds from various populations together with physiological stress related to their transport and confinement at a sales complex, may be associated with shedding and transmission of EHV-1 and -4. This prospective cohort study sampled 90 young Thoroughbreds consigned from eight farms, originating from three provinces representative of the South African Thoroughbred breeding demographic to a sales complex. Nasal swabs for quantitative real-time polymerase chain reaction (qPCR) assay to detect EHV-1 and -4 nucleic acid and blood samples for enzyme-linked immunosorbent assay for EHV-1 and -4 antibodies were collected from all horses on arrival and departure. Additional nasal swabs for qPCR were obtained serially from those displaying pyrexia and, or nasal discharge. Daily faecal samples were used for determination of faecal glucocorticoid metabolite (FGM) concentrations as a measurement of physiological stress and these values were modelled to determine the factors best explaining FGM variability. RESULTS : EHV-4 nucleic acid was detected in 14.4 % and EHV-1 from none of the animals in the study population. Most (93.3 %) and very few (1.1 %) of this population showed antibodies indicating prior exposure to EHV-4 and EHV-1 respectively. Pyrexia and nasal discharge were poor predictors for detecting EHV-4 nucleic acid. The horses’ FGM concentrations increased following arrival before decreasing for most of the remaining study period including the auction process. Model averaging showed that variation in FGM concentrations was best explained by days post-arrival and transport duration. CONCLUSIONS : In this study population, sales consignment was associated with limited detection of EHV-4 nucleic acid in nasal secretions, with most showing prior exposure to EHV-4 and very few to EHV-1. The physiological stress response shown by most reflected the combination of stressors associated with transport and arrival and these are key areas for future investigation into management practices to enhance health and welfare of young Thoroughbreds during sales consignment.Racing South Africa (Pty) Ltd, the Equine Research Centre and Departments of Companion Animal Clinical Studies and Production Animal Clinical Studies, Faculty of Veterinary Science, University of Pretoria, South Africa.http://www.biomedcentral.com/bmcvetresam201

An Equine Model for Vaccination against a Hepacivirus: Insights into Host Responses to E2 Recombinant Protein Vaccination and Subsequent Equine Hepacivirus Inoculation

Equine hepacivirus (EqHV) is the closest known genetic homologue of hepatitis C virus. An effective prophylactic vaccine is currently not available for either of these hepaciviruses. The equine as potential surrogate model for hepacivirus vaccine studies was investigated, while equine host responses following vaccination with EqHV E2 recombinant protein and subsequent EqHV inoculation were elucidated. Four ponies received prime and booster vaccinations (recombinant protein, adjuvant) four weeks apart (day −55 and −27). Two control ponies received adjuvant only. Ponies were inoculated with EqHV RNA-positive plasma on day 0. Blood samples and liver biopsies were collected over 26 weeks (day −70 to +112). Serum analyses included detection of EqHV RNA, isotypes of E2-specific immunoglobulin G (IgG), nonstructural protein 3-specific IgG, haematology, serum biochemistry, and metabolomics. Liver tissue analyses included EqHV RNA detection, RNA sequencing, histopathology, immunohistochemistry, and fluorescent in situ hybridization. Al-though vaccination did not result in complete protective immunity against experimental EqHV inoculation, the majority of vaccinated ponies cleared the serum EqHV RNA earlier than the control ponies. The majority of vaccinated ponies appeared to recover from the EqHV-associated liver insult earlier than the control ponies. The equine model shows promise as a surrogate model for future hepacivirus vaccine research

Detection of equine herpesvirus -4 and physiological stress patterns in young Thoroughbreds consigned to a South African auction sale

Commingling of horses from various populations, together with physiological stress associated with transport and confinement at a sales complex, may be associated with detection and transmission of equine herpesvirus type-1 (EHV-1) and -4 (EHV-4). This prospective cohort study aimed to investigate the currently undefined prevalence of EHV-1 and -4 in young Thoroughbreds at an auction sale in South Africa, and associations between clinical signs, physiological stress and viral detection. Ninety, two-year old Thoroughbreds (51 colts, 39 fillies) were consigned from eight farms and sampled at a South African auction sale. The horses were monitored for pyrexia and nasal discharge. Nasal swabs were collected for quantitative polymerase chain reaction (qPCR) assay to detect EHV-1 and -4 and faecal samples were collected for enzyme immunoassay (EIA) to determine faecal glucocorticoid metabolite (FGM) concentrations. EHV-4 nucleic acid was detected in some and EHV-1 nucleic acid in none of the population. Pyrexia and nasal discharge were poor indicators of EHV-4 status. Variation in FGM concentrations was best explained by transportation and preparation for auction. Peaks in EHV-4 detection and increases in FGM concentrations were identified shortly post-arrival and on the first day of auction. Temporal changes in FGM concentrations of horses from individual farms showed two distinct patterns: Pattern A (biphasic peaks) and Pattern B (single peak). It was concluded that sales consignment was associated with some EHV-4 nucleic acid detection and distinctive physiological stress patterns in this population of young Thoroughbreds.Dissertation (MSc)--University of Pretoria, 2014.tm2015Companion Animal Clinical StudiesMScUnrestricte

Stress steroid levels and the short-term impact of routine dehorning in female southern white rhinoceroses (Ceratotherium simum simum)

Rhinoceros populations in Africa are under severe threat as a result of surging poaching rates and risk-mitigation strategies are continuously adapted in an attempt to ensure the survival of the species. This study compared faecal glucocorticoid metabolite (fGCM) levels of two age classes of limited free-ranging female white rhinos with fGCM levels of adult free-ranging female white rhinos. Subsequently, fGCM alterations in the limited free-ranging animals were monitored following routine dehorning as a measure of the animals’ short-term physiological stress response. Baseline fGCM levels differed significantly between tested groups, with both free-ranging and limited free-ranging adult animals showing significantly higher fGCM levels compared with limited free-ranging juvenile females. In contrast, baseline fGCM levels did not differ significantly between limited free-ranging and free-ranging adult individuals. Routine dehorning procedures resulted in a short-term stress response expressed by a significant increase in fGCM levels 48 h post-dehorning, with stress steroid levels returning to pre-dehorning concentrations 72 h after the procedure.Keywords: faecal glucocorticoid metabolites, non-invasive hormone monitoring, physiological stress, South Afric

No Evidence of Mosquito Involvement in the Transmission of Equine Hepacivirus (Flaviviridae) in an Epidemiological Survey of Austrian Horses

Prevalence studies have demonstrated a global distribution of equine hepacivirus (EqHV), a member of the family Flaviviridae. However, apart from a single case of vertical transmission, natural routes of EqHV transmission remain elusive. Many known flaviviruses are horizontally transmitted between hematophagous arthropods and vertebrate hosts. This study represents the first investigation of potential EqHV transmission by mosquitoes. More than 5000 mosquitoes were collected across Austria and analyzed for EqHV ribonucleic acid (RNA) by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Concurrently, 386 serum samples from horses in eastern Austria were analyzed for EqHV-specific antibodies by luciferase immunoprecipitation system (LIPS) and for EqHV RNA by RT-qPCR. Additionally, liver-specific biochemistry parameters were compared between EqHV RNA-positive horses and EqHV RNA-negative horses. Phylogenetic analysis was conducted in comparison to previously published sequences from various origins. No EqHV RNA was detected in mosquito pools. Serum samples yielded an EqHV antibody prevalence of 45.9% (177/386) and RNA prevalence of 4.15% (16/386). EqHV RNA-positive horses had significantly higher glutamate dehydrogenase (GLDH) levels (p = 0.013) than control horses. Phylogenetic analysis showed high similarity between nucleotide sequences of EqHV in Austrian horses and EqHV circulating in other regions. Despite frequently detected evidence of EqHV infection in Austrian horses, no viral RNA was found in mosquitoes. It is therefore unlikely that mosquitoes are vectors of this flavivirus

Active equine parvovirus‐hepatitis infection is most frequently detected in Austrian horses of advanced age

Background: Equine parvovirus-hepatitis (EqPV-H) research is in its infancy. Information regarding prevalence, geographical distribution, genetic diversity, pathogenesis and risk factors enhances understanding of this potentially fatal infection. Objectives: Determining the prevalence of EqPV-H in Austrian equids. Investigating factors increasing probability of infection, liver-associated biochemistry parameters, concurrent equine hepacivirus (EqHV) infection and phylogenetic analysis of Austrian EqPV-H variants. Study design: Cross-sectional study. Methods: Sera from 259 horses and 13 donkeys in Austria were analysed for anti-EqPV-H VP1-specific antibodies by luciferase immunoprecipitation system (LIPS) and EqPV-H DNA by nested polymerase chain reaction (PCR). Associations between infection status, sex and age were described. Glutamate dehydrogenase (GLDH), gamma-glutamyl transferase (GGT), bile acids and albumin concentrations were compared between horses with active infection and PCR-negative horses. PCR targeting partial EqPV-H NS1 was performed and phylogenetic analysis of Austrian EqPV-H variants was conducted. Complete coding sequences (CDS) of four Austrian variants were determined by next-generation sequencing (NGS) and compared with published sequences. Results: Horses' EqPV-H seroprevalence was 30.1% and DNA prevalence was 8.9%. One horse was co-infected with EqHV. Significantly, higher probability of active EqPV-H infection was identified in 16- to 31-year-old horses, compared with 1- to 8-year-old horses (P = 0.002; OR = 8.19; 95% CI = 1.79 to 37.50) and 9- to 15-year-old horses (P = 0.03; OR = 2.96; 95% CI = 1.08 to 8.17). Liver-associated plasma parameters were not significantly different between horses with active infection and controls. Austrian EqPV-H variants revealed high similarity to sequences worldwide. No evidence of EqPV-H was detected in donkeys. Main limitations: Equids' inclusion depended upon owner consent. There was only one sampling point per animal and the sample of donkeys was small. Conclusions: EqPV-H antibodies and DNA are frequently detected in Austrian horses, without associated hepatitis in horses with active infection. The risk of active EqPV-H infection increases with increasing age. Phylogenetic evidence supports close relation of EqPV-H variants globally, including Austrian variants