Healthy Horse Research Articles

Development of the equine vibration arthrometry system (EVAS) for the study of equine lameness

Friction caused at different articular surfaces in horses’ joints can produce various vibration signals. In this study, we collected and analyzed the articular vibration signals in the fetlock joints of a healthy horse, an aged horse, and a horse with laminitis using the equine vibration arthrometry system (EVAS). The data obtained from the EVAS enabled the researchers to easily understand the condition of the horses’ inner joints and to differentiate between the joints of healthy limbs and those of diseased limbs with musculoskeletal disorders. Furthermore, we also developed mathematical algorithms to analyze the data from the EVAS in this study. We identified two periodic waveform cycles for each horse’s step in the time domain. The negative waveform cycle first appeared at each aged horse’s step. The root-mean-square (RMS) values of both the positive and negative waveform cycles were significantly larger at the first periodic waveform in the aged horse. In contrast, the positive waveform cycle first appeared in each healthy horse’s step and the RMS values of the positive waveform cycle were significantly larger in the healthy horse. We also measured the energy of the articular vibration signals of the healthy and aged horses in the first and second periodic waveform cycle during each horse’s step. By analyzing and comparing articular vibration signals in these horses, we were able to determine which of the horses had a musculoskeletal disorder. EVAS is a simple, convenient and non-invasive method of identifying articular problems in equine joints.

Equestrian chilblain: another outdoor recreational hazard

Herein, we describe two cases and review 14 cases of equestrian chilblain or 'equestrian cold panniculitis' in the literature. The first, a 23-year-old healthy female horse trainer, presented with burning nodular swelling on her lateral thighs. The second was a 34-year-old healthy woman with recurrent nodular eruption on the lateral thighs after horseback riding in the winter. Physical examination of both patients revealed erythematous to violaceous nodules with eczema craquelé-like changes. Laboratory workup for systemic and autoimmune connective tissue disease was negative. Punch biopsies from both patients showed a superficial and deep perivascular and periadnexal lymphocytic infiltrate with focal extension into the subcutaneous fat. Parakeratosis, subtle spongiosis and increased pandermal interstitial mucin were also present. Previously reported cases generally showed a similar clinical course and similar histopathologic findings. In contrast, our cases revealed increased pandermal interstitial mucin, resembling tumid lupus erythematosus. We aim to better characterize the histopathologic findings of equestrian chilblain and discuss its relationship to other cold-induced skin injuries and autoimmune connective tissue disease, namely lupus erythematosus.

Development and evaluation of a SYBR Green real-time RT-PCR assay for evaluation of cytokine gene expression in horse

Cytokine secretion is one of the main mechanisms by which the immune system is regulated in response to pathogens. Therefore, the measurement of cytokine expression is fundamental to characterizing the immune response to infections. Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) is widely used to measure cytokine mRNA levels, but assay conditions should be properly evaluated before analyzing important equine infections through relative quantification of gene expression. The aim of this study was to develop and evaluate a set of RT-qPCR assays for a panel of the most common cytokines in horses involved in innate and adaptive immune responses. Eight cytokines (interleukin (IL)-1β, IL-2, IL-4, IL-10, IL-12, TNFα, IFNβ and IFNγ) and a housekeeping gene (β-actin) were detected and amplified with the same annealing temperature in a SYBR Green RT-qPCR assay of samples of mitogen-stimulated peripheral blood mononuclear cells from a healthy horse and whole blood from a horse infected with African horse sickness virus. The method gave good efficiency for all genes tested, allowing quantification of relative expression levels. These SYBR Green RT-qPCR assays may be useful for examining cytokine gene expression in horses in response to exposure to economically important pathogens.

Theileria equi merozoite antigen-2 interacts with actin molecule of equine erythrocyte during their asexual development

Theileria equi is a tick-transmitted intraerythrocytic protozoan parasite in equids. Equine merozoite antigen (EMA)-1 and EMA-2 of T. equi have been identified as immunodominant proteins co-expressed on the surface of extra-erythrocytic merozoites. Additionally, only the EMA-2 is shed into the cytoplasm of infected erythrocyte or inside the erythrocytic membrane during their early developmental stage. In this study, we initially performed West-Western blot analysis on Triton X-100-insoluble erythrocytic skeleton collected from a healthy horse, using a glutathione S-transferase (GST)-tagged recombinant EMA-1t or EMA-2t of T. equi. The results indicated positive interactions of actin and band 4.1 molecules in the equine erythrocytic skeleton only with the recombinant EMA-2t. Subsequently, we carried out GST pull-down assay using the recombinant antigens (as above) against solubilized lysate of equine erythrocytic skeleton, and confirmed the co-precipitation of actin molecule with EMA-2t, but not with the EMA-1t. The interaction of EMA-2 with host erythrocytic actin indicated its role in the pathobiology of T. equi infection within host erythrocytes.

Plasma pharmacokinetics and pharmacodynamics of alfaxalone in neonatal foals after an intravenous bolus of alfaxalone following premedication with butorphanol tartrate

ObjectiveTo determine the pharmacokinetics and pharmacodynamics of the neurosteroid anaesthetic, alfaxalone, in neonatal foals after a single intravenous (IV) injection of alfaxalone following premedication with butorphanol tartrate. Study designProspective experimental study. AnimalsFive clinically healthy Australian Stock Horse foals of mean ± SD age of 12 ± 3 days and weighing 67.3 ± 12.4 kg. MethodsFoals were premedicated with butorphanol (0.05 mg kg−1 IV) and anaesthesia was induced 10 minutes later by IV injection with alfaxalone 3 mg kg−1. Cardiorespiratory variables (pulse rate, respiratory rate, direct arterial blood pressure, arterial blood gases) and clinical signs of anaesthetic depth were evaluated throughout anaesthesia. Venous blood samples were collected at strategic time points and alfaxalone plasma concentrations were assayed using liquid chromatography-mass spectrometry (LC/MS) and analysed by noncompartmental pharmacokinetic analysis. ResultsThe harmonic, mean ± SD plasma elimination half life (t½) for alfaxalone was 22.8 ± 5.2 minutes. The observed mean plasma clearance (Clp) and volume of distribution (Vd) were 19.9 ± 5.9 mL minute kg−1 and 0.6 ± 0.2 L kg−1, respectively. Overall, the quality of the anaesthetic inductions and recoveries was good and most monitored physiological variables were clinically acceptable in all foals, although some foals became hypoxaemic for a short period following recumbency. The mean durations of anaesthesia from induction to first movement and from induction to standing were 18.7 ± 7 and 37.2 ± 4.7 minutes, respectively. ConclusionsThe anaesthetic protocol used provided a predictable and consistent plane of anaesthesia in the five foals studied, with minimal cardiovascular depression. In foals, as in the adult horse, alfaxalone has a short elimination half life. Clinical relevanceAlfaxalone appears to be an adequate anaesthetic induction agent in foals and the pharmacokinetics suggest that, with continuous infusion, it might be suitable to provide more prolonged anaesthesia. Oxygen supplementation is recommended.

Comparison of the Sedative and Hemodynamic Effects of Acepromazine and Promethazine in the Standing Horse

The objective of this study was to compare the sedative and peripheral hemodynamic effects of acepromazine (ACP) and promethazine (PTZ) in the standing healthy horse. Nine healthy Warmblood horses randomly received either intravenous ACP at 0.1 mg/kg or PTZ at 0.1, 0.2, and 0.3 mg/kg. A sedation score based on clinical examination was recorded, and systolic arterial blood pressure was noninvasively evaluated using a Doppler flow detector at the tail, just before and every 15 minutes until 60 minutes after drug injection. Hemodynamics of the median artery of the left forelimb was studied using Doppler ultrasonography just before and 45 minutes after injection of the drug, which allowed calculation of surface (SURF), diameter (DIAM), and circumference (CIRC) of the vessel and peak systolic velocity (PSV), end diastolic velocity (EDV), mean velocity (MV), volumetric flow (VF), and resistivity index (RI) of the blood flow. Regardless of the dose used, PTZ had lesser sedative and hypotensive effects than ACP at 0.1 mg/kg and did not induce significant variations in SURF, DIAM, CIRC, PSV, EDV, MV, VF, and RI of the studied standing horses. Conversely, the vasodilatory properties of ACP were illustrated by a significant increase in SURF, DIAM, CIRC, PSV, EDV, MV, and VF and a significant reduction of the RI. Unlike ACP, PTZ did not induce alterations on the morphology of the Doppler waveform. PTZ appears to have less sedative and peripheral vasodilator effects than ACP, thus it could be safer than ACP in patients suffering from hypotension.

No evidence of horizontal infection in horses kept in close contact with dogs experimentally infected with canine influenza A virus (H3N8)

BackgroundSince equine influenza A virus (H3N8) was transmitted to dogs in the United States in 2004, the causative virus, which is called canine influenza A virus (CIV), has become widespread in dogs. To date, it has remained unclear whether or not CIV-infected dogs could transmit CIV to horses. To address this, we tested whether or not close contact between horses and dogs experimentally infected with CIV would result in its interspecies transmission.MethodsThree pairs of animals consisting of a dog inoculated with CIV (108.3 egg infectious dose50/dog) and a healthy horse were kept together in individual stalls for 15 consecutive days. During the study, all the dogs and horses were clinically observed. Virus titres in nasal swab extracts and serological responses were also evaluated. In addition, all the animals were subjected to a gross pathological examination after euthanasia.ResultsAll three dogs inoculated with CIV exhibited clinical signs including, pyrexia, cough, nasal discharge, virus shedding and seroconversion. Gross pathology revealed lung consolidations in all the dogs, and Streptococcus equi subsp. zooepidemicus was isolated from the lesions. Meanwhile, none of the paired horses showed any clinical signs, virus shedding or seroconversion. Moreover, gross pathology revealed no lesions in the respiratory tracts including the lungs of the horses.ConclusionsThese findings may indicate that a single dog infected with CIV is not sufficient to constitute a source of CIV infection in horses.

Vitreal IgM Autoantibodies Target Neurofilament Medium in a Spontaneous Model of Autoimmune Uveitis

Although the presence of IgG autoantibodies in the vitreous of spontaneous cases of equine recurrent uveitis (ERU) has been demonstrated, the potential role of IgM reactivities during ERU pathogenesis remains unexplored. The purpose of this study was to examine the presence of IgM autoantibodies in vitreous specimens of ERU-affected horses and to test their binding specificity to intraocularly expressed proteins. To test IgM autoantibody responses to retinal tissue, vitreous samples of eye-healthy controls and ERU patients were analyzed via two-dimensional Western blot analysis with equine retinal tissue as an antigen source. A candidate protein, the peptide neurofilament medium (NF-M), was identified via mass spectrometry and validated via enzyme-linked immunosorbent assay. Immunohistochemistry for NF-M expression was performed on healthy and ERU-affected retinal sections. Whereas autoreactivity was never detected in the healthy vitreous samples, NF-M was specifically targeted by vitreal IgM autoantibodies in 44% of the ERU cases. Vitreal anti-NF-M IgG was detected in only 8% of the ERU samples, pointing to a persistent IgM response. In healthy horse retina, NF-M was located in the retinal ganglion cells and their processes, with additional staining in the outer plexiform layer. NF-M expression in ERU-affected retinas decreased considerably, and the remaining expression was limited to the nerve fiber layer. Intraocular anti NF-M IgM autoantibodies occur with high prevalence in vitreous of spontaneous autoimmune uveitis cases. The IgM dominated response may indicate a thymus-independent response to NF-M and merits further investigation in ERU, as well as in its human counterpart, autoimmune uveitis.

Looking for an Ideal Horse: Rider Preferences

ABSTRACTThe aims of the present study were as follows: (1) to identify the particular characteristics of a horse that are important to riders, and (2) to examine which of these properties are significant for different categories of riders. The participants (197; 78.7% females) were surveyed using the Ideal Horse Questionnaire (IHQ), which contains items that are categorized as “physical make-up,” “behavior during riding and handling,” “training level,” and “maintenance.” “Behavior during riding and handling” (43.6% of respondents) and “physical make-up” (32.8%) were selected most frequently as the most important category, followed by “training level” (16.9%) and “maintenance” (6.7%). The responses for the questionnaire were subjected to factor analysis (FA). The first four factors of the FA, labeled “ease of handling and maintenance/sociability,” “jumping suitability,” “dressage suitability,” and “challenge” accounted for 13.7%, 11.7%, 8.6%, and 5.9% of the variance, respectively. The highest values on the 0–3 scale were obtained for items related to “ease of handling and maintenance/sociability” (mean ± SD = 1.97 ± 0.48) and “jumping suitability” (1.86 ± 0.62), followed by “dressage suitability” (1.64 ± 0.41) and “challenge” (1.20 ± 0.62). All participants desired a manageable, healthy and non-problematic horse. Men preferred a horse that was a good jumper and would challenge the rider. People who rarely rode, beginner to intermediate riders, non-competitors and all but show jumpers were not interested in a horse with a high “jumping suitability.” Only more experienced riders expected the horse to possess high capabilities and extensive training in dressage. Challenging animals were desirable to men but were avoided by leisure riders. The present study shows that ease of handling and maintenance, along with “jumping suitability,” are crucial attributes of a riding horse. Furthermore, various levels of experience and preferred equestrian activity are associated with different horse requirements.

Analysis of serum proteins electrophoresis in clinically healthy miniature Caspian horse

The Iranian miniature Caspian horse is the most ancient domestic breed of horse; it is the likely ancestor of all modern breeds of hot-blooded horses. Blood serum protein analysis has not been sufficiently studied in miniature Caspian horses. Since serum electrophoresis may be an important preclinical tool for diagnosis of equine diseases, evaluation of serum protein fractions in miniature Caspian horses using acetate cellulose strips was undertaken. In addition, serum protein concentrations were also examined. Blood samples were collected from 30 clinically healthy horses with and without anticoagulant. Using cellulose acetate electrophoresis (CAE), serum samples were fractionated and absolute and relative concentrations from each sample calculated. From 30 samples, 26 samples (86.6%) had five protein fractions and four samples (13.4%) had four fractions. Samples with five fractions were a common pattern (χ 2, P 0.05). Absolute values of albumin, α1, α2, β, and γ globulin were 2.9 ± 0.24, 0.97 ± 0.19, 0.96 ± 0.24, 1.2 ± 0.36, and 1.3 ± 0.29 g/dl, respectively. Mean albumin to globulin ratio was 0.67. In conclusion, analysis of serum proteins in the miniature Caspian horse using CAE showed five protein fractions including albumin, α1, α2, β, and γ globulin.

Expression of inflammation-associated genes in circulating leukocytes collected from horses with gastrointestinal tract disease

To investigate whether expression of inflammation-associated genes in leukocytes from horses with gastrointestinal tract (GIT) diseases correlated with the type of disease and outcome. 10 healthy horses and 50 horses with GIT disease. A blood sample was collected from each healthy horse or horse with GIT disease (during admission to the hospital). Leukocytes were isolated, diluted to a standard concentration, and frozen until RNA extraction. Expression of 14 genes associated with inflammation was quantified by use of a real-time quantitative reverse transcription PCR assay. Results were grouped by GIT disease type and disease outcome for comparison. Horses with GIT disease had colic of unknown etiology (n = 8 horses), GIT inflammation or strangulation (19), or nonstrangulating GIT obstruction (23). Among the 45 horses receiving treatment, 38 were discharged from the hospital, and 7 died or were euthanized. Compared with healthy horses, horses with colic of unknown etiology had similar gene expression. Significant differences in expression of the interleukin-8, leukocyte-selectin molecule, matrix metalloproteinase-9, platelet-selectin molecule, mitochondrial superoxide dismutase, Toll-like receptor 4, and tumor necrosis factor-A genes were detected between healthy horses and horses with GIT disease. Significant differences in expression of the interleukin-1 receptor antagonist, interleukin-8, leukocyte-selectin molecule, matrix metalloproteinase-9, platelet-selectin molecule, mitochondrial superoxide dismutase, Toll-like receptor 4, and tumor necrosis factor-A genes were detected among healthy horses and horses grouped by disease outcome. Inflammatory gene expression in leukocytes of horses with GIT disease appeared to be related to disease pathogenesis and prognosis.

Methicillin-Resistant Staphylococci Isolated from Healthy Horses and Horse Personnel in Italy

Methicillin-resistant staphylococci (MRS) were isolated from nasal swabs of 56 of 159 (35.2%; 95% confidence interval [CI]: 27.9-43.2%) healthy horses. Two nasal swabs were collected from each horse; 43 of 159 (27%; 95% CI: 20.5-34.8%) of the cohort were colonized by MRS strains in 1 nostril, while in the remaining 13 of 159 (8.2%; 95% CI: 4.6-13.9%), different or identical MRS strains were isolated in both nostrils. Of the 29 humans in close contact with the horses tested, 4 (13.8%; 95% CI: 4.5-32.6%) were found to be carriers of MRS. All isolates were coagulase negative with the exception of 2 coagulase-positive MRS strains, Staphylococcus aureus and Staphylococcus pseudintermedius, both isolated from horses. To assay the methicillin resistance, a susceptibility test to oxacillin with standardized disk diffusion method, a PBP-2a latex agglutination test, and a methicillin resistance gene (mecA) polymerase chain reaction assay were performed. Pulsed-field gel electrophoresis patterns of isolates from horses and humans in close contact with the horses revealed similarity. The results suggest evidence of transmission between animals, from animals to humans, and vice versa.

Acid-base balance parameters and a value of anion gap of arterial and venous blood in Małopolski horses

The comparative study of the acid-base balance (ABB) parameters has been performed on 20 clinically healthy mature Małopolski horses. An arterial blood sample from the facial artery and a sample of venous blood from the external cervical vein were colected from each animal. In the samples tested, the blood pH, pCO2, tCO2, HCO3-, concentration of Na+, K+, Cl-, and a value of the anion gap were determined. The difference among pCO2, tCO2, and HCO3- in both samples tested was statistically significant, whereas the pH of the arterial blood and the pH of the venous blood did not differ significantly. The anion gap in both types of blood did not differ significantly. 1) ABB parameters such as pCO2, HCO3-, and tCO2 determined in the arterial and venous blood of the Małopolski horses differ from each other significantly. 2) In spite of the lack of the differences between pH of the arterial and venous blood, the ABB parameters in horses should be determined in the arterial blood, because the comparative study performed proves that the analysis of the ABB parameters determined for the venous blood of a healthy horse may lead to a wrong diagnosis of the compensated respiratory acidosis. 3) The mean value of anion gap in horses aged 8-12 years amounts to 20.9 mmol/l for the arterial blood and 19.93 for the venous blood; the difference between the two values is not statistically significant.

Neuropathogenic and non-neuropathogenic genotypes of Equid Herpesvirus type 1 in Argentina

Infection with Equid Herpesvirus type 1 (EHV-1) leads to respiratory disease, abortion, and neurological disorders in horses. Molecular epidemiology studies have demonstrated that a single nucleotide polymorphism (A2254/G2254) in the genome region of the open reading frame 30 (ORF30), which results in an amino acid variation (N752/D752) of the EHV-1 DNA polymerase, is significantly associated with the neuropathogenic potential of naturally occurring strains. In order to estimate the prevalence of the EHV-1 neuropathogenic genotype in our country, we analyzed the ORF30 genome region of Argentinean EHV-1 isolates. The study was carried out by real time allelic discrimination PCR in 90 equine EHV-1-positive samples, being 89 from 54 cases of abortion outbreaks (two of which were in association with neurological disease) and one from the respiratory tract of a healthy horse in training. Our results indicate that 7% (4/54) of the abortion outbreaks studied were induced by the neuropathogenic (G2254) genotype of EHV-1 and 50% (2/4) of them were associated with simultaneous neurological disease. This information emphasizes the necessity to extreme the hygienic and preventive measures to diminish EHV-1 infections and consequently reduce the risk of epizootic neurological disease as has been recently observed in other countries.

Neurectomy of the deep branch of the lateral plantar nerve can cause neurogenic atrophy of the muscle fibres in the proximal part of the suspensory ligament(M. interosseous III)

Neurectomy of the deep branch of the right lateral plantar nerve was performed on a single healthy mature horse. Six weeks after surgery, the horse was subjected to euthanasia and both hind suspensory ligaments harvested. The cross sectional area of the muscular part of the proximal part of the suspensory ligament was measured and assessed for morphological abnormalities in a blinded fashion. There was a clear difference in cross sectional area of the muscular part between treated and control ligament and there was profound neurogenic atrophy of the muscular fibres in the treated ligament.

Leptospira seroprevalence and associations between seropositivity, clinical disease and host factors in horses

BackgroundA cross-sectional study was carried out to determine the seroprevalence of different serovars of Leptospira spp. and their association with clinical disease and host factors in Swedish horses.MethodsSera from 2017 horses brought to equine clinics during 1997–98 were investigated. The sera were examined by microscopic agglutination test for the presence of antibodies against the following L. interrogans serovars: Bratislava strain Jez, Icterohaemorrhagiae strain Kantorowicz and Pomona strain Pomona and also L. kirschneri sv Grippotyphosa strain Duyster and L. borgpetersenii sv Sejroe strain M 84. Host factors, disease factors, season, pasture access and outdoor confinement variables were analysed with respect to seropositivity to sv Bratislava and Icterohaemorrhagiae. Multivariable logistic regression was used to model seropositivity to sv Bratislava and Icterohaemorrhagiae (seroprevalence > 8%).ResultsThe seroprevalence, at a cut-off 1:100, were for sv Bratislava (16.6%), Icterohaemorrhagiae (8.3%), Sejroe (1.2%), Pomona (0.5%) and Grippotyphosa (0.4%). In the multivariable analysis, it was demonstrated that seroprevalence increased with age for sv Bratislava and Icterohaemorrhagiae. For sv Bratislava the seasons April – June and October – December and for sv Icterohaemorrhagiae October – December had higher seroprevalences than other seasons. Horses not used for racing had higher levels of seropositivity to sv Bratislava. Furthermore, horses with respiratory problems as well as horses with fatigue had higher levels of seropositivity to sv Bratislava. Ponies and coldbloods, and horses with access to pasture, had lower seroprevalence for sv Icterohaemorrhagiae. Healthy horses had lower seroprevalence for sv Icterohaemorrhagiae, than non-healthy horses.ConclusionThere was no significant association between clinical signs and disease and positive titres to sv Bratislava (except for the association between respiratory problems and fatigue and seropositivity to sv Bratislava). The results suggest that horses with increasing age and exposed to factors associated with outdoor life had an increased seroprevalence for sv Bratislava, indicating that horses get infected from outdoor and/or are exposed to shedding from other horses (management dependent). For sv Icterohaemorrhagiae, management possibly plays a role as ponies and coldbloods as well as healthy horses had lower seroprevalence. Overall, the age of the horse should be taken into consideration when evaluating the titre as the average healthy horse has a higher titre than a young horse.

Molecular Characterization of Rhodococcus equi from Horse-Breeding Farms by Means of Multiplex PCR for the vap Gene Family

This study evaluated the molecular characteristics of Rhodococcus equi isolates obtained from horses by a multiplex PCR assay that amplifies the vap gene family (vapA, -B, -C, -D, -E, -F, -G, and -H). A total of 180 R. equi isolates were studied from four different sources, namely healthy horse feces (112), soil (12), stalls (23), and clinical isolates (33) from horse-breeding farms. The technique was performed and confirmed by sequencing of amplified vap gene family controls. Thirty-two (17.8%) of the R. equi isolates were positive for the vapA gene and carried at least three other vap genes. All 147 isolates from equine feces, stalls, and soil failed to demonstrate any genes associated with virulence-inducing proteins. About 32 (97.0%) out of the 33 clinical equine isolates tested positive for the multiplex PCR assay for the vap gene family. They demonstrated six molecular profiles: 100% featured the vapA, vapD, and vapG genes, 86.6% vapF, 76.6% vapH, 43.3% vapC, 36.6% vapE, and none vapB. The most frequent molecular profile was vap A, -D, -F, G, and -H, where this profile was present in 37.5% of the strains. Moreover, there was no molecular epidemiological pattern for R. equi isolates that uniquely mapped to each horse-breeding farm studied. Our proposed technique allows the identification of eight members of the vap gene family (vapA, B, -C, -D, -E, -F, -G, and -H). It is a practical and efficient method of conducting clinical and epidemiological studies on R. equi isolates.

Comparison of cytologic and histologic evaluations of the conjunctiva in the normal equine eye

To describe the cells observed in conjunctival brush cytology (CBC) from normal horses and compare these findings with conjunctival structural histology so as to understand which cells are recovered from CBC. This study was divided into three parts. (1) Conjunctival brush smears were collected from 20 healthy horses on both eyes and a differential count on 300 cells was carried out on May Grünwald-Giemsa (MGG) smears. (2) A similar protocol was used for whole eyes from five horses obtained rapidly after death from a slaughterhouse. The eyes were then assessed for conjunctival histology. (3) Cytobrush smears were collected from five healthy horses. Smears were examined after MGG or periodic acid Schiff (PAS) staining. The differential cell count showed a majority of deep and intermediate epithelial cells with very few superficial and goblet cells in both eyes. A stratified columnar to cuboidal epithelium was observed on nearly the whole surface of the conjunctiva. A stratified squamous type was observed at the palpebral and bulbar edges. Areas with highest mucus cell indices were found from the nasal to the temporal edge of the equine inferior conjunctiva in the upper palpebral segment near the fornix and in a part of the nasal fornix. In MGG smears no mucus cells were identified; however, they were numerous in PAS smears (22.6% +/- 11) and were mostly cylindrical cells (42.5% +/- 14.4 PAS positive). Cytobrush smears in the healthy horse are characterized by a majority of polyhedral and cylindrical cells and a few squamous cells. The cylindrical cells may be mucous cells and probably originate from the main stratified columnar to cuboidal epithelium.

IgE and IgG antibodies in skin allergy of the horse

In horses, allergies have been characterized by clinical signs and/or intradermal (i.d.) allergen testing. Our aim was to find the first direct evidence that immunoglobulin E (IgE) mediates equine allergy. In addition, we tested the hypothesis that immediate skin reactions in horses can also be mediated by IgG. Anti-IgE affinity columns were used to purify IgE from serum of one healthy horse and three horses affected with summer eczema, an allergic dermatitis which is believed to be induced by Culicoides midges. A modified Prausnitz-Küstner experiment was performed in four clinical healthy horses by i.d. injection of the purified serum IgE antibodies. The following day, Culicoides allergen was injected at the same sites. Skin reactions were not observed in response to allergen alone, and in two horses after stimulation at any previous IgE injection site. However, the other two horses showed an immediate skin reaction at the previous injection sites of IgE obtained from allergic horses. In addition, purified monoclonal antibodies to various equine immunoglobulin isotypes were injected i.d. into six healthy horses. Immediate skin reactions were observed in response to anti-IgE (6/6 horses) and anti-IgG(T) injections (5/6 horses). The specificities of both antibodies for IgE and IgG(T), respectively, were confirmed by enzyme linked immunosorbent assays. The results provide the first direct evidence that IgE mediates classical Type-I allergy in horses and plays a major role in the pathogenesis of summer eczema. The data also suggest that IgG(T) can bind to skin mast cells and might contribute to clinical allergy.

Isolation of Equine Herpesvirus-5 from Blood Mononuclear Cells of a Gelding

Horses are commonly infected by herpesviruses, but isolation of equine herpesvirus-5 (EHV-5) has only infrequently been reported. We describe the isolation and characterization of a strain of EHV-5 from the blood mononuclear cells of a healthy adult horse in California. The virus was initially identified by EHV-5 specific polymerase chain reaction (PCR), and it caused lytic infection of cultured rabbit kidney cells only after repeated serial passage. Virions with characteristic herpesvirus morphology were readily demonstrated in cell culture lysate by transmission electron microscopy. A portion of the glycoprotein B gene of this strain of EHV-5 had 99% identity to the published EHV-5 sequence, and it was clearly distinguishable from other EHV (1-4) by virus-specific PCR assays. Prevalence of EHV-5 infection in a group of young racehorses was estimated at 64% using the EHV-5 specific PCR on nasopharyngeal secretions.