Equine Population Research Articles

UnsaddlingStreptococcus equiinfection of horses

Infection with Streptococcus equi, which forms abscesses in the lymph nodes of the head and neck in horses, is endemic in almost all countries around the world. The identification and isolation of horses with fever, an early sign of disease, is critical to minimising the number of horses affected and the severity of an outbreak, while the identification and treatment of persistently infected ‘carrier’ horses can reduce the risk of recurrent outbreaks and transmission between equine populations. Rapid diagnostic testing plays a key role in the identification of infected horses, which can then be isolated before the development of acute disease or treated to clear persistent infection. Vaccination can also be used to reduce the number of horses that become infected and the severity of their ensuing disease. This review describes the tools available to veterinarians and the journey towards the development and launch of a multi-component fusion protein vaccine that does not trigger positive diagnoses with any of the available diagnostic tests for strangles. The use of vaccination, alongside conventional methods of biosecurity and diagnostic testing, has the potential to unsaddle S. equi, reducing the number of strangles outbreaks and enhancing the health of horses.

Taylorella asinigenitalis: raising awareness of its importance and presence in equine and asinine populations.

Taylorella equigenitalis has long been recognised as a causative agent of contagious equine metritis, but practitioners may be less familiar with Taylorella asinigenitalis, which has been identified more recently. Here, Abel Dorrego, Consuelo Serres and Fatima Cruz-Lopez of the Universidad Complutense de Madrid describe T asinigenitalis and report the findings of a survey they carried out in donkeys in Spain.

Validation of an indirect in-house ELISA using synthetic peptides to detect antibodies anti-gp90 and gp45 of the equine infectious anaemia virus.

Equine infectious anaemia (EIA) is controlled by the identification of seropositive animals. The official diagnostic method is the agar gel immunodiffusion (AGID) test, which detects antibodies against a viral core protein (p26). Although AGID is inexpensive and specific, the report of results takes considerable time and the test has low analytical sensitivity. To validate our in-house indirect ELISAgp90/45 , following the World Organization of Animal Health (OIE) criteria. Test validation. Synthetic peptides gp90 and gp45 were used as antigens in ELISAgp90/45 . Tests used for validation, calibration and linear working operating range, analytical and diagnostic sensitivity and specificity, repeatability and reproducibility were assessed by comparing them with the AGID test and using 1844 equine sera grouped into five different panels. We were able to replace the National References Sera with our Internal Reference Sera. ELISAgp90/45 had acceptable repeatability and reproducibility. Analytical sensitivity of the ELISAgp90/45 was 800 times greater than that of AGID test for positive sera and 400 times greater for weak positive sera. ELISAgp90/45 also showed optimal analytical specificity, since no cross-reactivity was detected with antibodies against other equine viruses. One sample was positive by AGID test and negative by ELISAgp90/45. ELISAgp90/45 was performed using 243 EIA positive and 878 negative equid sera, and showed a diagnostic sensitivity of 99.59% [CI 97.73%-99.99%] and a diagnostic specificity of 90.32% [CI 88.17%-92.19%], compared to AGID test; thus, it was demonstrated to be a robust test. Samples were derived from naturally infected equid populations showing heterogeneous clinical states: therefore, their status was uncertain and some horses were sampled more than once. The AGID test may not be the most useful gold standard. ELISAgp90/45 is a useful tool for the diagnosis of EIAV infection and meets validation requirements established by the OIE.

Detecting paroxysmal atrial fibrillation from normal sinus rhythm in equine athletes using Symmetric Projection Attractor Reconstruction and machine learning

BackgroundAtrial fibrillation (AF) is a common cardiac arrhythmia in both human and equine populations. It is associated with adverse outcomes in humans and decreased athletic performance in both populations. Paroxysmal atrial fibrillation (PAF) presents with intermittent, self-terminating AF episodes, and is difficult to diagnose once sinus rhythm resumes.ObjectiveWe aimed to detect PAF subjects from normal sinus rhythm equine electrocardiograms (ECGs) using the Symmetric Projection Attractor Reconstruction (SPAR) method to encapsulate the waveform morphology and variability as the basis of a machine learning classification.MethodsWe obtained ECG signals from 139 active equine athletes (120 control, 19 with a PAF diagnosis). The SPAR method was applied to 9 short (20-second) ECG strips for each subject. An optimal SPAR feature set was determined by forward feature selection for input to a machine learning model ensemble of 3 different classifiers (k-nearest neighbors, linear support vector machine, and radial basis function kernel support vector machine). Imbalanced data were handled by upsampling the minority (PAF) class. A final subject classification was made by taking a majority vote over results from the 9 ECG strips.ResultsOur final cross-validated classification for a subject gave an accuracy of 89.0%, sensitivity of 94.8%, specificity of 87.1%, and receiver operating characteristic area under the curve of 0.98, taking PAF as the positive class.ConclusionThe SPAR method and machine learning generated a final model with high sensitivity, suggesting that PAF can be discriminated from short equine ECG strips. This preliminary study indicated that SPAR analysis of human ECG could support patient monitoring, risk stratification, and clinical decision-making.

The Gingiva of Horses With Pituitary Pars Intermedia Dysfunction: A Macroscopic Anatomical Evaluation.

Pituitary pars intermedia dysfunction (PPID) is a common neurodegenerative disease mainly in horses older than 15 years. The domestic equine population is following the same demographic change as that seen in humans; it is aging and veterinarians are asked to attend to geriatric horses more frequently. Common problems seen regularly in older equines are dental disorders and especially periodontal disease. As a systemic and endocrine disease, associated with delayed wound healing and impaired immune function, PPID should be considered before major dental treatment in aged equines is started. Possible negative effects of PPID on epithelial tissues could also affect the periodontium. Therefore, the aim of the present study was to identify gross changes in the gingiva associated with PPID. Fourteen horses with clinical signs of PPID and adenoma in the pituitary pars intermedia and 13 controls showing neither clinical signs nor PPID-associated histological changes in the pituitary gland were included. PPID-affected horses (26.9 ± 0.73 years) were significantly older than controls (20.0 ± 1.24 years). In the PPID-affected group, significantly more often an irregular and bulky appearance of the gingival texture was observed, as well as an irregular shape of the gingival margin. Furthermore, the sulcus gingivalis of cheek teeth frequently was deeper than 1 mm. These findings indicate a possible association between age, soft tissue alterations, and PPID and suggest a potential predisposition of PPID-affected horses for periodontal diseases.

Clostridioides difficile infection and One Health: an equine perspective.

Summary Clostridioides (Clostridium) difficile presents a significant health risk to humans and animals. The complexity of the bacterial–host interaction affecting pathogenesis and disease development creates an ongoing challenge for epidemiological studies, control strategies and prevention planning. The recent emergence of human disease caused by strains of C. difficile found in animals adds to mounting evidence that C. difficile infection (CDI) may be a zoonosis. In equine populations, C. difficile is a known cause of diarrhoea and gastrointestinal inflammation, with considerable mortality and morbidity. This has a significant impact on both the well‐being of the animal and, in the case of performance and production animals, it may have an adverse economic impact on relevant industries. While C. difficile is regularly isolated from horses, many questions remain regarding the impact of asymptomatic carriage as well as optimization of diagnosis, testing and treatment. This review provides an overview of our understanding of equine CDI while also identifying knowledge gaps and the need for a holistic One Health approach to a complicated issue.

Equine asthma syndrome: managing the environment

Equine asthma syndrome is arguably the most common non-septic lower respiratory condition diagnosed in horses. In a hospital environment, it is important to be aware of its clinical signs, especially as many horses with the condition will go unnoticed by their owners. Prompt identification can help reduce morbidity within the hospital. Focus on environmental management practices, including feed, bedding and ventilation are well known to reduce the prevalence of the disease within the equine population. This review provides practical advice to equine professionals operating within a hospital environment to minimise respirable particle concentration and the incidence of equine asthma syndrome. A checklist is provided to allow interactive discussion with owners at discharge into how best to reduce the clinical signs of equine asthma syndrome at home.

Advances in the understanding of laminitis and the role of nutrition in its management

Laminitis heavily impacts the equine population and, for this reason, has long been researched with respect to its prevention, causation, and management. Recent research has uncovered the importance of owner recognition in both the prevention and management of laminitis and highlighted this as a potential area for education. Furthermore, clearer definitions of the syndrome and its causation, along with risk of recurrence, should enable a more tailored approach to nutrition and feed management. Dissemination of current research into feeding practice should be seen as a key part of the ‘next steps’ in laminitis management and prevention. This review collates current recommendations with applications to practical feeding situations, and demonstrates that while there is information on what constitutes an appropriate diet ‘on paper’, the practicalities of providing this (particularly where forage is concerned), often represents a barrier to long-term use. It is in this area that future research should focus, to fully realise and benefit from the progress made to date.

Development of IgM-ELISA for diagnosis of recent infection of Japanese encephalitis virus in equines

Japanese encephalitis (JE) is a re-emerging mosquito borne disease, for which equines are most susceptible amongst all animals. Detection of specific immunoglobulin ‘M’ (IgM) is considered as an ideal way to diagnose recent JE virus infection in equines due to low virus load and short-term viremia. The present study was undertaken to develop a sensitive and specific recombinant NS1 protein based indirect IgM-ELISA and IgM capture (MAC) ELISA to diagnose recent infection of JEV in equines. Indirect IgM ELISA was standardized with relative diagnostic sensitivity and specificity of 100% and 88.5%, respectively. The validation of indirect IgM-ELISA in different laboratories revealed excellent reproducibility with Cohen's kappa value ranging between 0.84 and 1. The standardization of MAC ELISA was attempted using checker board titration method and non-specific binding of polyclonal anti-equine IgM capture antibody with anti-porcine IgG conjugate and with hyperimmune serum raised in swine against the antigen was observed. Hence, the MAC ELISA was standardized with monoclonal capture antibody; however, its diagnostic performance could not meet the satisfactory limit. Due to better sensitivity and less turnaround time, indirect IgM-ELISA was employed to screen 821 equine serum samples revealing 33.73% positivity of IgM antibodies against JEV in equine population of India. The high JEV sero-positivity warrants the need for vaccination in Indian equine population along with the demand for research focused towards anti-viral therapy. The indirect IgM-ELISA developed in the present study could be useful to diagnose acute or recent infection of JEV in equines as well as in sero-epidemiological studies.

ANÁLISIS GENEALÓGICO EN LA POBLACIÓN ABIERTA DE CABALLOS SILLA ARGENTINO PERTENECIENTES A LA POLICÍA NACIONAL DE COLOMBIA

This study aimed to evaluate population parameters, genetic diversity and identify the ancestors with the highest genetic contribution of an open population of Silla Argentino horses in Colombia using pedigree data. 508 horses born between 1998 and 2018 were evaluated, called reference population (RP). The ancestors of the RP were added, defining the total population (TP) whit 1,861 horses born since 1905. Population parameters were estimated, including inbreeding coefficient (F), effective population size (Ne), effective number of founders (fe), ancestors (fa), and founder genomes (fg) and generation interval (GIN). The GIN was 10.1 (TP) and 7.7 (RP) years. The estimate of F for the RP was 0.3%, indicating control in mattings. The Ne was 317 (RP) and 179 (TP). The probability of genes origin indicated values for fe, fa and fg of 386, 187 and 351 (TP), and 161, 64 and 56 (RP), respectively, evidencing a reduction of genetic variability in the RP. Half of the genetic variation in the equine population studied was explained by 40 ancestors. The results show loss of genetic diversity in the RP and the genetic contribution of the ancestors suggests the need to increase the number of stallions for breeding and guide mating to increase genetic diversity in the new generations of Silla Argentino horses of the Colombian National Police

Long-term adaptation following influenza A virus host shifts results in increased within-host viral fitness due to higher replication rates, broader dissemination within the respiratory epithelium and reduced tissue damage

The mechanisms and consequences of genome evolution on viral fitness following host shifts are poorly understood. In addition, viral fitness -the ability of an organism to reproduce and survive- is multifactorial and thus difficult to quantify. Influenza A viruses (IAVs) circulate broadly among wild birds and have jumped into and become endemic in multiple mammalian hosts, including humans, pigs, dogs, seals, and horses. H3N8 equine influenza virus (EIV) is an endemic virus of horses that originated in birds and has been circulating uninterruptedly in equine populations since the early 1960s. Here, we used EIV to quantify changes in infection phenotype associated to viral fitness due to genome-wide changes acquired during long-term adaptation. We performed experimental infections of two mammalian cell lines and equine tracheal explants using the earliest H3N8 EIV isolated (A/equine/Uruguay/63 [EIV/63]), and A/equine/Ohio/2003 (EIV/2003), a monophyletic descendant of EIV/63 isolated 40 years after the emergence of H3N8 EIV. We show that EIV/2003 exhibits increased resistance to interferon, enhanced viral replication, and a more efficient cell-to-cell spread in cells and tissues. Transcriptomics analyses revealed virus-specific responses to each virus, mainly affecting host immunity and inflammation. Image analyses of infected equine respiratory explants showed that despite replicating at higher levels and spreading over larger areas of the respiratory epithelium, EIV/2003 induced milder lesions compared to EIV/63, suggesting that adaptation led to reduced tissue pathogenicity. Our results reveal previously unknown links between virus genotype and the host response to infection, providing new insights on the relationship between virus evolution and fitness.

A new cluster of West Nile virus lineage 1 isolated from a northern goshawk in Spain

West Nile Virus (WNV; family Flaviviridae, genus flavivirus) is a zoonotic arbovirus worldwide spread. Its genetic diversity has allowed the definition of at least seven lineages, being lineages 1 and 2 the most widely distributed. Western Mediterranean region has been affected by WNV since decades. In Spain, WNV is actively circulating, provoking annual outbreaks in birds, horses and lately in humans. Lineage 1 is responsible for outbreaks that occurred in central and southern regions, while lineage 2 has been recently described in wild birds in north-eastern part of the country. During 2017 season, a disease outbreak in captive raptors was reported in southern Spain and WNV was isolated from a dead northern goshawk. Full genome sequencing was followed by phylogenetic analyses and analyses of the amino acidic substitutions. This strain, named Spain/2017/NG-b, highly differs from those which have been circulating both in Spain and in the neighbouring Mediterranean countries, constituting a new distinct group, tentatively classified in a newly defined cluster 7 within the WNV clade 1a, supporting a new, independent introduction of the virus in the Western Mediterranean region from an unknown origin. Besides, circumstantial evidence indicates that this emerging WNV strain could be behind the subsequent outbreak occurred nearby in horses. Overall, the reinforcement of surveillance programs, especially in wild birds, is essential to early detect the circulation of WNV and other related flaviviruses that could cause outbreaks in wild or domestic birds, equine and human populations.

What can equine surveillance learn from surveillance of other species?

Mary-Ellen Lloyd Tivey of the Royal Veterinary College, suggests that greater collaboration and encouraging engagement from horse owners and vets is key to strengthening scanning surveillance in equine populations.

Retrospective Analysis of Cause-of-Death at an Equine Retirement Center in the Netherlands Over an Eight-Year Period

Geriatric horses (>15 years) represent a substantial proportion of the equine population. The objective of this study was to (1) report on the cause of death in geriatric horses on a single retirement centre, (2) to evaluate the relationship between PPID status, and mortality rate. Data of 194 horses was collected over an eight-year period. This providing data for 80 geriatric cases of mortality. The average age of horses that died was 26.2 years (±4.8 years). The majority (95%) of mortalities were euthanised, most frequently due to colic (20%), old age (19%), and lameness (15%). Overall mortality rate was 16.3 per 100 horse years at risk (95% C.I. 15.2–17.5) for the total geriatric (>15 years) population. PPID had a prevalence of 48.8% (61 out of 125 horses). The results of this study demonstrate increasing mortality rates with increasing age in geriatric horses. The most common reasons for euthanasia were lameness, old age, and colic. The treatment of PPID does not result in an older age of death. The effect of treatment on the welfare status was not studied. Management, of both veterinarians and care taker, has an important role in maintaining health and welfare in geriatric horses. Findings of this study could be relevant to veterinary surgeons and stable managers providing care for geriatric horses.

Sero-positivity of Japanese Encephalitis Virus in Equine Population of India Using IgG ELISA: Unraveling the Need for Vaccination

Japanese encephalitis (JE) is a mosquito borne flaviviral zoonoses, causing fatal disease in equines and humans. JE is endemic in most of the states of India with occurrence of human cases every year. The horses are not vaccinated against JE in India and thus they are at more risk of acquiring the disease. Due to nonavailability of indigenously developed ELISA and high cost of imported kits, regular sero-surveillance is not being carried out to assess the true picture of JE virus in equine population of India. Therefore, a recombinant NS1 protein based indirect IgG ELISA was developed with the objective to assess the sero-positivity of JE virus in equine population of India. The diagnostic sensitivity and specificity of developed ELISA was 84.73% and 86.70%, respectively. The validation studies revealed good reproducibility of ELISA with kappa value ranging from 0.75 to 1 between the results of different laboratories. A total of 2,069 horse serum samples were screened using the developed ELISA and 401 samples were positive for IgG against JEV with an overall sero-positivity of 19.38% in equine population of India. A sero-positivity of 25.90% and 12.22% was recorded in Himachal Pradesh and Jammu-Kashmir, both hill states of North zone of India for the first time, revealing the spread of virus to the nonendemic parts of the country. The high sero-positivity of JE virus recorded in equine population warrants the need for initiation of vaccination of horses in India to prevent the morbidity and mortality.

Autumn de-worming

Autumn is traditionally a time when most horses are de-wormed with one or more products that are effective against tapeworms and both adult and larval roundworms. However, the increasing prevalence of resistance and availability of diagnostics should prompt scrutiny of whether blanket de-worming at this time of year is appropriate. If management is good and horses are at low-risk of clinical disease, then there may be no need to use anthelmintics. Serology provides a means of assessing exposure to both cyathostomins and tapeworms, but it has its limitations and does not provide a validated means of assessing the risk of parasitic disease. The results of serology, results of faecal worm egg counts performed in the population through the preceding grazing season, and knowledge of management practices in the population should all be considered when assessing whether the risk of the disease to the individual is such that it must take priority over the ever-increasing threat of resistance in the equine population.

Equine obesity and the role of the veterinary nurse

Equine obesity is defined as a medical disease in which excess body fat has accumulated to such an extent that it has an adverse effect on the general health of the horse. Obesity is a cause for concern, with one-third of the equine population in the UK being regarded as obese, although owner recognition of obesity in horses is an inherent problem, with many underestimating the body condition or weight of their horse. This is further complicated by the fact that with larger framed horses, or horses that are already overweight, assessing body condition is more difficult. There are a number of ways to assess body condition and the most practical means of regular assessment is body condition scoring, although this is regarded as subjective. As with many diseases and disorders, the cause of obesity is multifactorial. However, the most common reason for a horse to become obese is overfeeding, coupled with a lack of exercise. Obesity can be addressed with client education and veterinary nurses can provide advice on weight management programmes. However, these need to be tailored to the individual horse and owners need to recognise that they are entering into a long-term commitment.

Co-infection with Trypanosoma congolense and Trypanosoma brucei is a significant risk factor for cerebral trypanosomosis in the equid population of the Gambia

Trypanosomosis is a major cause of morbidity and mortality in working equids in The Gambia. Recently, a progressive, severe neurological syndrome characterised by a diffuse lymphoplasmacytic meningoencephalitis has been identified and associated with Trypanosoma brucei infection of the central nervous system. The pathogenesis of cerebral trypanosomosis is unclear and the clinical syndrome not well described. This observational cross-sectional study aimed to identify host and parasite related risk factors associated with the development of cerebral trypanosomosis and to describe the neurological syndrome associated with cerebral trypanosomosis. History, signalment, clinical and laboratory parameters were collected from 326 horses and donkeys presented to The Gambia Horse and Donkey Trust. Neurological derangements in affected animals were described. Species-specific polymerase chain reaction (PCR) for Trypanosoma congolense, Trypanosoma vivax and Trypanosoma brucei was performed. The associations between signalment, clinical and laboratory parameters and PCR results were assessed using multivariable logistic regression. The overall prevalence of trypanosomosis was 50 %, with infections dominated by T. congolense (44.1 %) and a lower intensity of T. brucei (7.4 %) and T. vivax (6.5 %). Overall, 54.8 % of neurological cases were PCR positive for trypanosomosis. Within the neurological sub-population prevalence remained similar to the whole population for T. congolense (48.4 %) and T. vivax (6.5 %); whilst the prevalence increased markedly for T. brucei (32.3 %). Co-infections were identified in 32.3 % of neurological cases. Donkeys typically presented with progressive cerebral dysfunction and cranial nerve deficits, whereas in horses a progressive spinal ataxia was predominant. Mortality in affected animals was high (82.4 %). The final multivariable model identified a significant association between body condition score ≤2 (OR 11.4; 95 % CI 4.6–27.9; P = <0.001), and T. congolense and T. brucei. coinfection (OR 20.6; 95 % CI 1.71–244.1; P = 0.016) with the presence of neurological deficits. This study has provided clinically relevant information confirming the link between T. brucei and neurological disease outbreak in the equid population of The Gambia, and crucially identified co-infection with T. brucei and T. congolense as a major risk factor for the development of neurological trypanosomosis. Further research is required to identify the epidemiology of co-infection in equidae of The Gambia, so that cerebral trypanosomosis can be better prevented in this vulnerable population.

16Years of breed management brings substantial improvement in population genetics of the endangered Cleveland Bay Horse.

The consequences of poor breed management and inbreeding can range from gradual declines in individual productivity to more serious fertility and mortality concerns. However, many small and closed groups, as well as larger unmanaged populations, are plagued by genetic regression, often due to a dearth in breeding support tools which are accessible and easy to use in supporting decision‐making. To address this, we have developed a population management tool (BCAS, Breed Conservation and Management System) based on individual relatedness assessed using pedigree‐based kinship, which offers breeding recommendations for such populations. Moreover, we demonstrate the success of this tool in 16 years of employment in a closed equine population native to the UK, most notably, the rate of inbreeding reducing from more than 3% per generation, to less than 0.5%, or that attributed to genetic drift, as assessed over the last 16 years of implementation. Furthermore, with adherence to this program, the long‐term impact of poor management has been reversed and the genetic resource within the breed has grown from an effective population size of 20 in 1994 to more than 140 in 2020. The development and availability of our BCAS for breed management and selection establish a new paradigm for the successful maintenance of genetic resources in animal populations.

Recent advancements in our understanding of equid gammaherpesvirus infections.

Equid gammaherpesviruses are ubiquitous and widespread in the equine population. Despite their frequent detection, their contribution to immune system modulation and the pathogenesis of several diseases remains unclear. Genetic variability and the combination of equid gammaherpesvirus strains a horse is infected with might be clinically significant. Initial gammaherpesvirus infection occurs in foals peripartum with latency then established in peripheral blood mononuclear cells. A novel EHV-5 study suggests that following inhalation equid gammaherpesviruses might obtain direct access to T and B lymphocytes via the tonsillar crypts to establish latency. EHV-5 is associated with equine multinodular pulmonary fibrosis, however, unlike with EHV-2 there is currently minimal evidence for its role in milder cases of respiratory disease and poor performance. Transmission is presumed to be via the upper respiratory tract with periodic reactivation of the latent virus in adult horses. Stress of transport has been identified as a risk factor for reactivation and shedding of equine gammaherpesviruses. There is currently a lack of evidence for the effectiveness of antiviral drugs in the treatment of equine gammaherpesvirus infections.