Hydrocortisone levels in the urine and blood of horses treated with ACTH.

An investigation was undertaken to demonstrate whether therapeutic treatment with ACTH raises hydrocortisone (cortisol) levels in horse urine above the limit (1000 ng/ml) established by the International Conference of Racing Authorities with the aim of controlling the abuse of cortisol and ACTH in equine sports. ACTH (200 iu) was administered i.m. to 3 Thoroughbred horses; urine and blood samples were collected at intervals afterwards and analysed by an immunoenzymatic system (ELISA) and HPLC-MS. To ascertain post exercise cortisol levels in untreated horses, 101 urine and 103 serum samples were taken from horses immediately after racing and analysed by ELISA. The peak urine level of cortisol, detected 8 h after ACTH administration, was around 600 ng/ml using either ELISA or HPLC-MS. The peak serum cortisol concentration was found to be around 250 ng/ml by ELISA, but consistently less by HPLC-MS. Mean cortisol levels in post race horses were 135.1+/-72.1 ng/ml in urine and 90.1+/-41.7 ng/ml in serum. High levels of the metabolite 20beta-dihydrocortisol in urine and the cortisol precursor 11beta-desoxycortisol in serum were found. The latter showed high cross-reactivity with cortisol on ELISA. In our experiment, treatment with ACTH 200 iu i.m. did not raise urinary cortisol levels above the 1000 ng/ml threshold proposed by the ICRA.     

Coagulation profiles of healthy Andalusian donkeys are different than those of healthy horses

Background: Coagulation disorders are frequently diagnosed, especially in hospitalized equidae, and result in increased morbidity and mortality. However, hemostatic reference intervals have not been established for donkeys yet. Objectives: To determine whether the most common coagulation parameters used in equine practice are different between healthy donkeys and horses. Animals: Thirty‐eight healthy donkeys and 29 healthy horses. Methods: Blood samples were collected to assess both coagulation and fibrinolytic systems by determination of platelet count, fibrinogen concentration, clotting times (prothrombin time [PT] and activated partial thromboplastin time [aPTT]), fibrin degradation products (FDP) and D‐Dimer concentrations. Results: PT and aPTT in donkeys were significantly (P < .05) shorter than those of horses. In contrast, FDP and D‐Dimer concentrations were significantly (P < .05) higher in donkeys than in horses. Conclusions and Clinical Importance: The coagulation parameters most commonly determined in equine practice are different in donkeys compared with horses. Thus, the use of normal reference ranges reported previously for healthy horses in donkeys might lead to a misdiagnosis of coagulopathy in healthy donkeys, and unnecessary treatments in sick donkeys. This is the first report of normal coagulation profile results in donkeys, and further studies are warranted to elucidate the physiological mechanisms of the differences observed between donkeys and horses.     

Effect of exercise on the immune response of young and old horses.

OBJECTIVE: To compare exercise-induced immune modulation in young and older horses. ANIMALS: 6 young and 6 aged horses that were vaccinated against equine influenza virus. PROCEDURE: Venous blood samples were collected for immunologic assessment before and immediately after exercise at targeted heart rates and after exercise for determination of plasma lactate and cortisol concentrations. Mononuclear cells were assayed for lymphoproliferative responses and incubated with interleukin-2 (IL-2) to induce lymphokine-activated killer (LAK) cells. Antibodies to equine influenza virus were measured. RESULTS: Older horses had significantly lower proliferative responses to mitogens than younger horses prior to exercise. Exercise caused a significant decrease in lymphoproliferative response of younger horses, but not of older horses. Activity of LAK cells increased slightly with exercise intensity in younger horses. Cortisol concentrations increased in both groups after exercise; younger horses had higher concentrations after exercise at heart rates of 180 and 200 beats/min than those of older horses. Plasma lactate concentrations increased with exercise intensity but there were no differences between older and younger horses. Older horses had lower antibody titers to equine influenza virus than younger horses. Exercise did not affect antibody titers. CONCLUSION: Although lymphoproliferative responses and antibody titers of older horses were less than those of younger horses, older horses were more resistant to exercise-induced changes in immune function, possibly because of lower cortisol concentrations. CLINICAL RELEVANCE: Stress and aging are known to affect immune function. Older horses had reduced immune function, but were more resistant to exercise-induced immune suppression than younger horses.     

Evaluation of a modified thrombelastography assay initiated with recombinant human tissue factor in clinically healthy horses

Background: Thrombelastography (TEG) is used to evaluate the viscoelastic properties of blood during clotting and provides a global assessment of hemostasis and clot lysis. TEG analysis initiated with recombinant human tissue factor (TF) has not been evaluated in clinically healthy horses. Objectives: The purpose of this study was to determine whether TEG results are affected by the time elapsed between sampling and analysis (storage time) of equine blood samples and to establish a preliminary equine reference interval for a modified TEG assay, using recombinant human TF to initiate coagulation. Methods: Citrated blood samples were obtained from 20 clinically healthy adult horses. Thirteen samples were stored for 30, 60, and 120 minutes at room temperature before TEG analysis. Coagulation was initiated by adding 20 μL of CaCl₂ to 330 μL of blood and 10 μL of diluted recombinant TF for a final dilution of 1:3600. Reaction (R) and clotting (K) times, angle (α), and maximum amplitude (MA) were compared between time points. A preliminary reference interval (minimum–maximum values) was determined using data from all 20 horses after 30 minutes of sample storage. Results: There was a significant effect of storage time on R, K, and α but not MA. Reference intervals were: R, 3.65–6.4 minutes; K, 1.8–5.45 minutes; α, 33.4–66.2°; MA, 41.2–64.1 mm; lysis at 30 minutes post‐MA (LY30), <2.75%; and lysis at 60 minutes post‐MA (LY60), 1.55–9.5%. Conclusions: TEG can be performed on equine citrated blood samples using recombinant human TF to activate clot formation. TEG parameters were significantly affected by storage time, suggesting an incomplete inhibition of coagulation in citrated blood.     

Interaction of saddle girth construction and tension on respiratory mechanics and gas exchange during supramaximal treadmill exercise in horses

OBJECTIVE: To determine the effect of girth construction and tension on respiratory mechanics and gas exchange during supramaximal treadmill exercise in horses. METHODS: Six healthy detrained Thoroughbred horses were exercised on a treadmill inclined at 10% at 110% VO2max. Horses were instrumented for respiratory mechanics and gas exchange studies, and data were recorded during incremental exercise tests. The animals were exercised for 2 min at 40% VO2max, and samples and measurements were collected at 1 min 45 sec. After 2 min, speed was increased to that estimated at 110% VO2max and data was collected at 45 sec, 90 sec and every 30 sec thereafter at this speed until the horses fatigued. Horses were run on three occasions with the same racing saddle and saddle packing but using two different girths, either an elastic girth (EG) or a standard canvas girth (SCG) which is nonelastic. A run with 5 kg tension applied to a standard canvas girth was the control for each horse, with additional runs at 15 kg using either the standard canvas girth or using the elastic girth. The runs were randomised and tensions applied were measured at end exhalation whilst at rest. RESULTS: Increasing girth tension was not associated with changes in respiratory mechanical or gas exchange properties. Although girths tightened to 15 kg tension had short run to fatigue times this was not found to be significantly different to girths set at 5 kg resting tension. Girth tensions declined at end exhalation in horses nearing fatigue. CONCLUSIONS: Loss in performance associated with high girth tensions is not due to alteration of respiratory mechanics. Loss in performance may be related to inspiratory muscles working at suboptimal lengths due to thoracic compression or compression of musculature around the chest. However, these changes are not reflected in altered respiratory mechanical or gas exchange properties measured during tidal breathing during supramaximal exercise. Other factors may hasten the onset of fatigue when horses exercise with tight girths and further studies are required to determine why excessively tight girths affect performance.     

Plasma ionized calcium and parathyroid hormone concentrations in horses after endurance rides

OBJECTIVE: To evaluate changes in plasma ionized calcium (Ca2+) and parathyroid hormone (PTH) concentrations in horses competing in endurance rides. DESIGN: Longitudinal clinical study. ANIMALS: 28 horses. PROCEDURE: Venous blood samples were obtained from horses before and after racing 80 km. Plasma pH and concentrations of Ca2+, PTH, inorganic phosphorus, albumin, lactate, and magnesium were measured. RESULTS: Overall, a significant decrease in mean (+/- SD) plasma Ca2+ concentration (from 6.44 +/- 0.42 to 5.64 +/- 0.42 mg/dl) and a significant increase in plasma PTH concentration (from 49.9 +/- 30.1 to 148.1 +/- 183.0 pg/ml) were found after exercise. Exercise also resulted in significant increases in plasma inorganic phosphorus, albumin, and lactate concentrations. No changes in plasma magnesium concentration or pH were detected after exercise. Plasma PTH concentration was not increased after exercise in 8 horses; in these horses, plasma PTH concentration decreased from 58.2 +/- 26.3 to 27.4 +/- 22.4 pg/ml, although plasma Ca2+ concentration was also decreased. CONCLUSIONS AND CLINICAL RELEVANCE: Plasma Ca2+ concentration was decreased after racing for 80 km, compared with values obtained before racing. In most horses, an increase in plasma PTH concentration that was commensurate with the decrease in plasma Ca2+ was detected; however, some horses had decreased plasma PTH concentrations.     

Tensions used on girths on thoroughbred racehorses

OBJECTIVE: To determine commonly used girth tensions in Thoroughbred racehorses in the Melbourne metropolitan area. DESIGN: A prospective industry survey. METHODS: Strappers were instructed to tension girths, using the same saddle, under-saddle packing and girth, sufficiently to hold the saddle for racing. These tensions were continuously recorded by an in-line load cell in the girth. Seventeen strappers from five stables participated in the study, which was conducted on 91 horses. RESULTS: Resting girth tensions varied considerably. The mean tension on inhalation was 13 +/- 0.4 kg. Male strappers recorded higher tensions than females for all indices measured. Age and girth size of the horse did not significantly influence results. CONCLUSION: This study confirms that there is no standard for application of a girth and saddle in the Thoroughbred racing industry and the range of tensions applied was large. Strappers were unable to reliably apply the same tensions between horses. Values recorded at the extremes of the range seem inappropriate for racing conditions as low values would result in slippage of the saddle and higher values may result in girth galls and possible reduction in athletic performance. Further work is necessary to determine the point at which girth tension affects performance.     

Effect of Competition on Nitric Oxide, Carbon Monoxide, and Cyclic Guanosine Monophosphate Levels in Training Horses

The aim of this study was to compare the effect of exercise on nitric oxide (NO), carbon monoxide (CO), and cyclic guanosyl monophosphate (cGMP) levels in jumping and dressage horses involved in competition. Blood samples were collected from the jugular vein of 100 horses involved in jumping or dressage competition at three time points: baseline at rest, on reaching the schooling but before exercise, and over a jump or dressage course. Fourteen healthy horses not involved in competition were used as control group. Exercise increased plasma CO concentration in both jumping and dressage horses, and this effect was more apparent in dressage horses. Exercise also increased NO plasma concentration in jumping horses, whereas it did not significantly modify NO plasma concentration in dressage horses. After exercise, plasma cGMP concentration was higher in both groups. Our results show that different signaling pathways are initially activated by exercise and that this activation is specific to the different modes of exercise. This information can be used to optimize warm-up and cool-down procedures for sport horses or to optimize training programs for equine athletes.     

Effect of Conditioning Horses Once,Twice, or Thrice a Week with High-Intensity Intermittent Exercise on v4

Intensive short intervals of exercise are used to condition horses for racing. In this study, the effect of exercising horses one, two, or three times a week during 6 weeks using two intervals with near-maximal speed over 100 m on v₄ (speed at which, under defined conditions, the blood lactate concentration reaches 4 mmol/L) and muscle thickness (MT) of supraspinatus and extensor carpi radialis was examined. Thoroughbreds (4-5 years of age) were exercised twice at near-maximal speed over 100 m, separated by a 10-minute period at walk, on dirt track during conditioning periods (CP) of 6-week duration. This exercise was undertaken once (six horses), twice (six horses), or thrice (five horses) a week during a CP. Before, every 2 weeks during, and 2 weeks after the CP, horses were subjected to a standardized exercise test to determine their v₄. Before and after CP, the MT of the extensor carpi radialis and the supraspinatus was examined by ultrasonography. There was no differential effect of the number of weekly exercises on v₄. Pooling the data of all horses, a decrease of v₄ was found. The decrease became evident after the sixth week of conditioning. MT did not change. The results indicate that the examined exercise protocols could negatively impact racing performance of horses.     

Exercised-induced pulmonary hemorrhage in polo and racing horses

Philippine polo and racing horses were examined for exercise-induced pulmonary hemorrhage after their competitive exercise. Exercise-induced pulmonary hemorrhage occurred in 11.1% of the polo horses and 64% of the racing horses. None of the horses had blood at the nostrils.     

Early exercise in the horse

Across all equestrian disciplines, the single largest reason for wastage is musculoskeletal injury. It is, therefore, of importance that management and competition structures are in place to optimize the development of the equine musculoskeletal system to minimize wastage.Data from other species and, in particular, humans have demonstrated the benefit of early exercise and the dire consequences of inactivity. The horse has evolved as a cursorial animal capable of covering up to 10 km/d within 9 days of birth. Yet, modern equine management systems restrict, rather than promote, early exercise. Foals were shown to have a positive response to early preweaning paddock exercise (greater cartilage health), and more recent work has demonstrated that exercise over and above that normally occurring with pasture-reared foals, introduced as early as 3 weeks old, had positive effects on the equine musculoskeletal system. The response of juvenile horses to additional exercise is because of the tissue being responsive to priming. Epidemiological data indicate that the window for tissue modification may still be open when the horse is a yearling and even as a 2 year old.However, the method in which the exercise is applied may be of as much importance as the timing of the stimuli. A recent prospective study of both Thoroughbred and Standardbred horses demonstrated that the horses that entered training as 2 year olds had longer and more successful racing careers than those that entered training later in life. It would appear that even the initial stages of training are enough to provide a positive stimulus, as horses first registered with a trainer at 2 years old had the same advantages as those that raced as 2 year olds.The physiological, clinical, and epidemiological data indicate that, rather than restrict exercise and the use of horses at a young age, we should realign expectations with the capability of the horses’ musculoskeletal system and evolutionary template to maximize orthopedic health.     

Intralesional injection of platelet-rich plasma followed by controlled exercise for treatment of midbody suspensory ligament desmitis in Standardbred racehorses

OBJECTIVE: To determine outcome of Standardbred racehorses with moderate to severe midbody suspensory ligament desmitis (MSD) treated by means of ultrasound-guided intralesional injection of a single dose of platelet-rich plasma (PRP) followed by a program of gradually increased exercise. DESIGN: Nonrandomized clinical trial. ANIMALS: 9 Standardbred racehorses. PROCEDURES: Following injection of PRP, horses were allowed a controlled, gradual return to exercise. Race records for the year prior to injury and for 3 consecutive years after horses returned to racing were reviewed. For comparison purposes, race records of 9 Standardbred racehorses with no history of MSD racing at the same time were also reviewed. RESULTS: All 9 horses with MSD returned to racing after treatment; median time to return to racing was 32 weeks. All 9 horses raced at least once during the first and second years after returning to racing, but only 5 raced during the third year. When number of starts, total earnings, and earnings per start were compared between case and comparison horses, the only significant differences were number of starts during the third year after case horses returned to racing and earnings per start during the first year after case horses returned to racing, with values being significantly lower for case horses than for comparison horses. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that horses with moderate to severe MSD treated by means of intralesional injection of a single dose of PRP followed by a program of gradually increased exercise had an excellent prognosis for returning to racing.     

Transendoscopic, laser-assisted ventriculocordectomy for treatment of left laryngeal hemiplegia in horses: 22 cases (1999-2005)

OBJECTIVE: To determine long-term effects of transendoscopic, laser-assisted ventriculocordectomy (LAVC) on airway noise and performance in horses with naturally occurring left laryngeal hemiplegia. DESIGN: Retrospective case series. ANIMALS: 22 horses with left laryngeal hemiplegia treated by means of LAVC. PROCEDURES: Medical records were reviewed and initial complaint, intended use of the horse, duration of abnormal airway noise, preoperative performance level, endoscopic findings, surgical procedure, postoperative treatment, and complications were recorded. Follow-up telephone interviews with owners and trainers were conducted to determine time for return to intended use, level of postoperative performance, and percentage reduction in airway noise. RESULTS: All horses were examined because of excessive airway noise; 10 (45%) had concurrent exercise intolerance. Left ventriculocordectomy was performed in all 22 horses; bilateral ventriculocordectomy (right ventriculocordectomy was done 1 year later) was performed in 1 horse (5%). Complications occurred in 3 (14%) horses. Twenty (91%) horses returned to their intended use. Excessive airway noise was eliminated after surgery in 18 (82%) horses; exercise intolerance improved postoperatively in 8 of 10 horses. Three racing Thoroughbreds returned to racing; 1 additional racehorse returned to racing but required a laryngoplasty 1 year later to continue racing. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that LAVC was an effective procedure for elimination of excessive airway noise and improvement of performance in horses with left laryngeal hemiplegia.     

Thromboelastography in equine medicine: Technique and use in clinical research

Thromboelastography (TEG) is a viscoelastic, whole blood‐based assay that integrates information from both the cellular and soluble components of coagulation, providing a global evaluation of the haemostatic system. This contrasts with the conventional coagulation assays (i.e. platelet count, prothrombin time [PT], activated partial thromboplastin time [aPTT] and fibrinogen concentration [FIB]), which only provide information about one component (e.g. clotting factors in the case of PT and aPTT) of the haemostatic process, requiring the combination of several assays for a complete evaluation of haemostasis. Thromboelastography is an old technology that has been used in human medicine for over 50 years. However, it is relatively new in veterinary medicine and has only been applied to horses in the last 5 years. Clinical applications in human medicine include diagnosis and monitoring of coagulopathies. Currently, extensive research is being carried out to expand the use of TEG in dogs and cats. Therefore, it is expected that the use of this technique will also further expand in horses in the near future. To date, the available studies in the equine species have evaluated TEG in healthy horses, horses with gastrointestinal disease, septic foals, horses with exercise‐induced pulmonary haemorrhage (EIPH) and a filly with Glanzmann's thrombasthenia. The main objective of this review is to introduce the TEG technique to equine clinicians, providing information on how the TEG functions, blood sample collection and processing, variables measured and their interpretations, normal reference values and areas of potential clinical application.     

Acute in vivo interactions of Helicobacter equorum with its equine host

REASONS FOR PERFORMING STUDY: A novel urease-negative Helicobacter species has been isolated from faecal samples of clinically healthy horses, but no information is available about the main sites of colonisation in the equine gastrointestinal tract nor is the pathogenic potential of this microorganism known. An experimental infection in horses was therefore carried out. METHODS: Four horses were infected with H. equorum strain CCUG 52199T and subjected to euthanasia at 10 (n = 2) and 30 days (n = 2) post inoculation. A fifth animal was inoculated with phosphate buffered saline and used as control. Gastrointestinal samples were examined histologically and bacteriologically. These samples, as well as faecal material collected at regular intervals, were also subjected to PCR analysis. RESULTS: All horses remained clinically healthy and no specific macroscopic lesions were identified, nor were there any microscopic changes. H. equorum-DNA was detected in the faeces during the whole experiment in all infected animals but not in the negative control. Sites of colonisation were caecum, colon and rectum. CONCLUSIONS: H. equorum is able to colonise the equine lower bowel and is excreted in faeces without apparent pathology. No association between the presence of the organism and gastrointestinal disease was demonstrated.     

Resting concentrations of cardiac troponin I in fit horses and effect of racing

ObjectivesTo determine normal resting values for cardiac troponin I (cTnI) in healthy Standardbred, Thoroughbred and Warmblood horses and investigate if racing has an influence on cTnI concentrations.BackgroundMeasuring cTnI concentrations in plasma is the gold standard for detecting myocardial injury in humans. Cardiac troponin I is highly conserved between species and has gained interest as a marker for cardiac injury in horses. Increased levels of cTnI have been reported in association with endurance and short-term strenuous exercise on a treadmill in horses. However, the effect of true racing conditions has not yet been reported.Animals, materials and methodsBlood samples for analysis of cTnI concentrations in plasma were collected from 67 Standardbred racehorses, 34 Thoroughbred racehorses and 35 Warmblood dressage horses at rest. Blood samples were also collected prior to and after racing in 22 Standardbred racehorses and 6 Thoroughbred racehorses.ResultsAll horses except one had resting plasma cTnI concentrations <0.022 μg/L. Mild increases in cTnI concentrations were seen in some horses 1–2 h after the race (1/17 Standardbreds and 2/6 Thoroughbreds) as well as 10–14 h after the race (4/21 Standardbreds and 1/6 Thoroughbreds).ConclusionsResting cTnI concentrations in horses are low but mildly elevated cTnI concentrations may be detected in some horses 1–14 h after racing. These findings could be of importance when evaluating horses with suspected cardiac disease that recently have performed hard exercise.     

Comparison of 4 blood storage methods in a protocol for equine pre-operative autologous donation

OBJECTIVE: To compare viability of equine whole blood stored by 4 different methods, and to establish optimal storage protocols for an equine autologous blood donation program. STUDY DESIGN: In vitro study of stored equine whole blood. Animals- Six healthy adult horses. METHODS: Blood from each horse was collected into 4 different containers: glass bottles containing acid-citrate-dextrose solution (ACD), plastic bags containing ACD, citrate-phosphate-dextrose (CPD), and CPD with supplemental adenine (CPDA-1). Blood was stored for 5 weeks and sampled at 2-day intervals. Standard hematologic and biochemical variables were evaluated, and adenosine-5-triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) concentrations were measured and normalized to total hemoglobin content. RESULTS: Plasma hemoglobin, % hemolysis, lactate, potassium, ammonia, and lactate dehydrogenase (LDH) increased, whereas glucose concentration and pH decreased in all stored blood over 5 weeks. There was a temporal increase in hemolysis with all storage methods, but the increase was greatest in glass bottles. Lactate and ammonia were highest in CPD and CPDA-1 samples, indicating more active red blood cell (RBC) metabolism. 2,3-DPG concentrations decreased during storage, but were optimally preserved with CPDA-1. ATP concentrations were significantly higher for blood stored in CPDA-1, and were lowest in glass bottles. CONCLUSIONS: Hematologic and biochemical values measured for blood stored in CPDA-1 are suggestive of improved RBC viability compared with other storage methods. With the exception of ATP, results from stored equine blood were similar to those reported for other species. CLINICAL RELEVANCE: Commercial CPDA-1 bags appear to be the optimal storage method for equine whole blood.     

Dynamic respiratory endoscopy of Standardbred racehorses during qualifying races

Reason for performing study: Examination of the equine upper airway during racing has not previously been documented. Objective: To describe the feasibility and appearance of the upper airways by overground respiratory endoscopic examination during racing conditions. Methods: Overground videoendoscopic examinations were performed on 46 Standardbred racehorses during qualifying races. Examined horses' speeds were recorded throughout the race with a portable GPS device. Results: The procedure did not interfere with performance as there were no significant differences in race times between races in which horses were examined with the endoscope in place and prior unexamined races. Airway obstructions during or after the race were documented in 21 horses. Most previously reported causes of upper airway obstruction were observed; surprisingly bilateral ventro‐medial arytenoid displacement (VMAD; n = 5) was seen during exercise as frequently as dorsal displacement of the soft palate (DDSP). Although DDSP (n = 10) was the most common diagnosis made, many displacements (n = 5) occurred after the race. Horses that demonstrated DDSP after the race had slower speeds than unaffected horses during the race. Conclusions: Racing endoscopy permits the diagnosis of upper airway obstructions without affecting performance. The occurrence of DDSP immediately after exercise may be clinically relevant. During racing VMAD may be an important anomaly. Potential relevance: Racing endoscopy could be used to correlate the sensitivity of diagnostic endoscopy during race‐training or treadmill examination. The pathogenesis and significance of VMAD deserves further investigation.     

Serum amyloid A protein (SAA) in horses: objective measurement of the acute phase response

A sensitive and precise immunoassay for equine serum amyloid A protein (SAA) was established and used to determine, for the first time, the circulating concentration of this protein in health and disease. As in other species, equine SAA was present only at trace levels in healthy animals but behaved as an extremely sensitive and rapidly responding acute phase reactant following most forms of tissue injury, infection and inflammation, objectively reflecting the extent and activity of disease. Measurements of SAA should make a significant contribution to diagnosis and management of viral and bacterial infection in horses, and routine serial assays could provide an objective criterion for monitoring prospectively the general health of horses in training and racing.     

Equus caballus papillomavirus‐2 (EcPV‐2): An infectious cause for equine genital cancer?

Reasons for performing study: The aetiology of genital squamous cell carcinoma (SCC) in horses remains unknown, but the similarity to the disease in man, for which papillomavirus infection has been shown to be a causal factor, requires to be investigated in horses. Hypothesis: One or more novel papillomaviruses cause equine genital SCC and its associated premalignant lesions. Methods: DNA was extracted from samples of equine genital SCC and performed rolling circle amplification, in order to identify closed circular DNA viral genomes within the samples. The amplified DNA was subcloned and sequenced and the DNA sequence compared to that of other papillomavirus genomes. Using PCR primers developed from these genomic DNA sequences, studies were then carried out in order to identify the frequency at which the viral DNA could be identified in equine genital cancer samples from horses in both the UK, Australia and Austria. Finally, in situ hybridisation using specific probes developed from this DNA sequence were used to confirm the presence of the viral RNA sequences in the neoplastic cells in these lesions. Results: The full length genome of a novel papillomavirus species was characterised from the equine genital SCC tissue and termed Equus caballus papillomavirus‐2 (EcPV‐2). Viral DNA and RNA was identified in the genital tumour samples, but not in the adjacent histologically normal tissue. EcPV‐2 DNA could not be identified in equine ocular or nasal carcinomas or within the scrotal skin or in most smegma samples obtained from tumour‐free horses. Sequencing of amplicons, generated from the archived equine genital tumours, identified variations within E1 and E6 on DNA and predicted protein level. Conclusions: A novel papillomavirus, EcPV‐2, is likely to play a causal role in the pathogenesis of equine genital epithelial tumours. Potential relevance: Identification of a papillomavirus causal for genital carcinomas in horses may lead to development of a vaccine that could be used to prevent this serious disease in horses. This would be analogous to man, where vaccination against oncogenic papillomavirus species is currently being used to help prevent cervical cancer.