Masa-aki Oikawa

Effect of passive immunization against inhibin on FSH secretion, folliculogenesis and ovulation rate during the follicular phase of the estrous cycle in mares.

Physiological roles of inhibin in mares were investigated by means of passive immunization using an antiserum to inhibin that had been raised in a castrated goat. Eight mares were given an intravenous injection of either 100 mL (n = 4) or 200 mL (n = 4) of inhibin antiserum 4 d after a single intramuscular injection of PGF2 alpha on Day 8 after ovulation, 4 control mares were treated with 100 mL castrated goat serum in the same manner. Jugular vein blood samples were collected after treatment with the serum until 192 h post treatment. Follicular growth and ovulations were monitored by ultrasound examination at 24-h intervals. The ability of the inhibin antiserum to neutralize the bioactivity of equine inhibin was examined in vitro using a rat pituitary cell culture system. Suppression of secretion of FSH from cultured rat pituitary cells by equine follicular fluid was reversed by the addition of increasing doses of the inhibin antiserum, thereby indicating its bioactivity. Plasma levels of FSH and estradiol-17 beta were higher in mares treated with the inhibin antiserum. The ovulation rate was significantly higher in mares treated with antiserum (100 mL = 3.75 +/- 0.63; 200 mL = 4.50 +/- 0.65) than in control mares (1.25 +/- 0.25). These results demonstrate that inhibin is important in regulating FSH secretion and folliculogenesis in mares. They also show that neutralization of the bioactivity of inhibin may become a new method for the control of folliculogenesis and ovulation rate in mares.

Pathology of equine respiratory disease occurring in association with transport

Summary Eight young thoroughbred horses, taken 1858 km by road (travelling time, 41 h), were exmanined toassess the pathological nature of respiratory disease associated with transport. Three of the horses showed clinical abnormalities including pyrexia, coughing, leucocytosis and neutrophilia after the first 20 h of transportation. Endoscopical examination of the trachea revealed exacerbation of airway inflammation as a result of transport in two of the three affected horses. A consistent finding in the affected horses was focal serous neutrophilic pneumonia affecting the cranio-ventral portion of the caudal lung lobe with a propensity to affect the right lung. Streptococcus equi subspecies zooepidemicus was isolated from the pneumonic areas, in which corresponding bacterial antigens were identified immunohistochemically. Viral cultures from the pneumonic lesions proved negative for respiratory viruses. It is suggested that transport predisposes the upper respiratory tract and the lower airways to invasion by the bacterium, with episodic pyrexia and acute pneumonia.

Pathology of equine pneumonia associated with transport and isolation of Streptococcus equi subsp. zooepidemicus.

Seven horses that died of pneumonia associated with transport yielded Streptococcus equi subsp. zooepidemicus (S.z.) from their pulmonary lesions. These lesions were divisible roughly into two types, serous haemorrhagic pneumonia and multiple foci of coagulative necrosis, which were considered to reflect a temporal difference in the process of lesion formation. Immunohistologically, S.z. antigen was detected in both types of lesion. Acute necrotic lacunar tonsillitis was considered to play an important role in the onset of the pneumonia.

Effect of restructuring of a racetrack on the occurrence of racing injuries in thoroughbred horses

Summary One of the race courses belonging to the Japan Racing Association was found to have defects in its geometrical design which predisposed racehorses to injury. This racetrack was reconstructed to eliminate these defects. This involved widening of the 3rd and 4th corners, and constructing a gradual curve in the straight stretch between these corners. Furthermore, the composition of the track was changed, and inclines and declines were constructed on the flat stretches. This remodeling of the track decreased the speed of the horses throughout the course, and the incidence of injuries was significantly reduced. Our findings indicate that remodeling of racetracks is effective for reducing the occurrence of racing injuries.

Surfactant proteins in bronchoalveolar lavage fluid of horses: assay technique and changes following road transport

An enzyme-linked immunosorbent assay (ELISA) was developed for equine surfactant proteins SP-A and SP-D in bronchoalveolar lavage fluid (BALF). Anti-equine SP-A or SP-D monoclonal antibodies (mAb) were produced by hybridoma technology, purified by the antibody purification reagent, and analysed by Western blotting analysis. The immunoreaction (two-site sandwich ELISA) with a mAb, peroxidase-labelled mAb and BALF sample was carried out simultaneously and analytical recovery and precision were assayed. Six mAb for SP-A and four mAb for SP-D were successfully cloned in limiting dilution to monoclonality. These mAb were reacted with equine SP-A or SP-D on Western blotting analysis. For SP-A, a combination of solid-phase TA08 and horseradish peroxidase (HRP)-conjugated WA28 was found to be more sensitive than other combinations, gave a good dose response and was capable of measuring 0.78 to 100 ng of protein/mi. For SP-D, a combination of solid-phase TD13 and HRP-conjugated WD19 was found to be more sensitive than other combinations, had a good dose response and was capable of measuring 0.78 to 200 ng of protein/ml. The assay was used to determine the effect of 41 hours of road transport on the concentrations of SP-A and SP-D in the BALF of 30 horses. The concentrations of SP-A and SP-D decreased by 55 per cent and 36 per cent, respectively, decreases similar to the decrease in phosphatidylglycerol concentration previously reported by the authors.

Epidemiology of racing injuries in Thoroughbred racehorses with special reference to bone fractures: Japanese experience from the 1980s to 2000s

ABSTRACT This report describes the descriptive epidemiology of racing fractures that occurred from the 1980s to 2000s on racetracks of the Japan Racing Association (JRA). The incidence of racehorse fractures during flat racing was approximately 1–2%. Fractures occurring during a race are more likely to occur in a forelimb. Fractures mostly occur at the third and fourth corners of oval tracks and on the home stretch. They also occur more frequently at the time of changing the leading limb. Comparison of the incidence of racing fracture between before and after reconstruction of the geometrical configuration of a racetrack revealed that there was an outstanding reduction in the number of serious fractures in the year before and after reconstruction. It was postulated that the improvement in racing time, possibly influenced by reconstructing the geometrical configuration of the racetrack, was connected to the reduction in the number of fractures. Of non-biological race- and course-related factors, type of course (dirt or turf), track surface condition, differences between racecourses, and racing distance significantly influence racing time. By using an instrumented shoe, vertical ground reaction forces (VGRFs) on the forelimb during galloping and the relationships between a rough dirt and woodchip track surface and a smooth dirt and woodchip surface were measured. Relating the incidence of racing fractures with track conditions in general showed that track surface has significant effects on the incidence of fracture, with the incidence of fractures increasing as track conditions on dirt worsen and a tendency for the incidence of fractures to decrease as track conditions on turf worsen. It seems probable that track condition in general may affect the incidence of fracture. The incidence of fracture in horses during both racing and training decreased as the years progressed.

Histopathological characteristics of endometrosis in thoroughbred mares in Japan: results from 50 necropsy cases.

ABSTRACT Uteri from 50 necropsied nonpregnant Japanese Thoroughbred brood mares (1–30 years of age) were investigated to clarify the histopathological characteristics of endometrosis in Japanese Thoroughbred mares and the distribution pattern of endometrosis lesions in the uterus as a whole. Endometrosis was observed in all animals over 6 years of age and in all of the 21 mares aged over 12 years of age. The affected mares showed elastofibrosis of arteries, veins and lymphatic vessels in the uterine wall, atrophy of the uterine smooth muscle layers and hyperplasia of collagen fibers among the smooth muscle fascicles of the myometrium, in addition to pathomorphologic features of endometrosis such as stromal endometrial fibrosis accompanied by endometrial atrophy, periglandular fibrosis and reduction of uterine glands. The severity of the histopathological changes increased with advancing age. Lymphatic vessels with elastofibrosis showed marked lymph congestion, leading to lymphatic edema. With increasing age, the extent of the distribution of these lesions tended to expand from focal to diffuse involvement of the entire uterus. Based on these findings, we speculate that aging plays a role in the pathogenesis of endometrosis; circulatory disturbances due to intrauterine angiosis or angiopathy, particularly reduction of the arterial blood supply and disturbance of venous drainage, resulting in a reduction of lymphatic drainage (lymphatic edema), are closely related to the onset and progression of endometrial fibrosis and myometrial atrophy with fibroplasia may result in myometrial hypofunction during the peri-implantation or puerperal period.

Morphology of equine allantochorion at the tip of the pregnant horn.

The morphology of the equine allantochorion at the tip of the pregnant horn was studied in the membranes of 14 mares. The findings in the allantochorion at the tip of the pregnant horn were of two types; one was growth retardation (hypoplastic villi, tunica adventitia of the vessels resembling embryonal connective tissue and the extended spaces of remnants of the extraembryonic coelom), the other was placental hypoxia or ischaemia (parakeratosis, stratified squamous metaplasia, necrosis of the trophoblasts, thickening of the basement membrane and fibrous hyperplasia of villous stroma). It seems likely that hypoplastic villi are caused by disordered formation of the microcotyledon accompanying the progression of pregnancy.

Arterionecrosis of the equine mesentery in naturally occurring endotoxaemia.

This report describes the mesenteric arteriolar lesions in a Thoroughbred racehorse with endotoxaemia due to colic. The vascular lesions consisted of a striking loss of medial smooth muscle cells, associated with granular cell debris derived from necrosed muscle cells, plasma insudation, erythrocyte infiltration and the deposition of a fibrinoid substance (fibrinoid degeneration) in the entire arterial wall, possibly produced by the infiltration of blood components through endothelial cell junctions into the arterial wall. The morphology of the mesenteric arteriolar necrosis closely resembled that seen in experimental equine endotoxaemia and in horses that died from colic; it also resembled that of Shiga toxin-induced arteriolar lesions in oedema disease of swine and of the arterionecrosis in human cerebral arteries that may lead to hypertensive intracerebral haemorrhage.

ENTHESOPATHY OF THE RADIAL TUBEROSITY IN TWO THOROUGHBRED RACEHORSES

Two cases of enthesopathy of the radial tuberosity in Thoroughbred racehorses are described. Soft X-ray pictures revealed separated bony fragments at the anterior aspect of the radial tuberosity, resembling the lesions of Osgood-Schlatter disease in children. Osgood-Schlatter lesions result from detachment of a portion of the apophysis of the tibial tuberosity. However, in the affected horses, the detached bony fragments consisted of cortical bone tissue composed of trabeculae with osteons similar to lamellar bone, the main component of the radial tuberosity. Tendon fibrils were inserted into the anterior parts of the detached fragments, and scar tissue filled the gap between the posterior part of the fragments and the radial tuberosity. The bone fragments may have originated from an avulsion fracture at the site of attachment of the tendinous portion of the biceps brachii muscle to the bone tissue of the radial tuberosity, a site at which high-tensile force is repeatedly exerted by muscle contraction.