Detection and effects on platelet function of anti-platelet antibody in mule foals with experimentally induced neonatal alloimmune thrombocytopenia.

Horse mares carrying mule foals were immunized during the last trimester of pregnancy with whole acid-citrate-dextrose-anticoagulated donkey blood to experimentally induce neonatal alloimmune thrombocytopenia. Thrombocytopenia occurred in the neonatal mule foals born to immunized horse mares within 24 hours after ingestion of their dams' colostrum. Mule foals born to mares not immunized with donkey blood did not develop thrombocytopenia. These findings suggest that antibodies may have been directed against a donkey platelet antigen present in the mule foals but not present in their dams. The objectives of this study were to determine whether anti-platelet antibody could be detected in mule foals with experimentally induced neonatal alloimmune thrombocytopenia, to identify any platelet proteins recognized by serum antibody in these foals, and to determine if platelet function was altered by sera from these mule foals. An indirect enzyme-linked immunosorbent assay demonstrated significantly higher absorption at 1:200 of platelet-bindable immunoglobulin G in serum from thrombocytopenic mule foals, compared with nonthrombocytopenic mule foals. Sera from thrombocytopenic and nonthrombocytopenic mule foals produced similar binding patterns in western immunoblots with donkey platelet proteins separated on sodium dodecyl sulfate polyacrylamide gels. Maximal platelet aggregation and relative slope of aggregation in response to collagen were significantly inhibited after incubation with sera from thrombocytopenic mule foals. These results suggest that mule foals with induced alloimmune thrombocytopenia have serum antibodies that bind to platelets and may compete with collagen binding sites to impair platelet aggregation.

Use of immunohistochemical methods for diagnosis of equine pythiosis.

Excisional biopsy specimens were received from 8 horses with lesions suspected of being caused by Pythium sp. Pythium sp was isolated from all specimens. An indirect peroxidase technique was applied to formalin-fixed tissues from the biopsy material and there was distinct staining of hyphal cell walls in all specimens. Sections containing hyphae, other than those of Pythium sp, failed to stain. The indirect peroxidase technique could be used for confirmation of suspected cases of pythiosis.

Evaluation of a technique for measurement of gamma-glutamyltranspeptidase in equine urine.

gamma-Glutamyltranspeptidase (GGT) activity in equine urine was measured, using an assay developed for use with serum and was found to be reproducible. The GGT activity was measured in samples prepared by serial dilution of exogenous GGT with equine urine, and the activity was determined to be linear between 21 IU/L and 407 IU/L. The behavior of exogenously added GGT was compared in equine serum and urine. The enzyme behaved similarly in both fluids. The GGT activity was measured in serum and urine samples after storage at -20, 4, and 25 C for 24 and/or 72 hours. Enzyme activity decreased after storage at all temperatures and times, but the decrease was greatest and most variable at -20 C. The urine GGT: urine creatinine ratio was calculated on serial urine samples collected over 24 hours. Although the urine GGT activity and creatinine concentration had significant variation, the ratio remained constant. The urine GGT: urine creatinine ratio was calculated from random urine samples collected from 27 clinically healthy adult horses and was found to be 10.52 +/- 4.78 (mean +/- SD).

Microculture method for mixed lymphocyte cultures in the horse.

A miniaturized method for the mixed lymphocyte culture test in the horse is described. The test is performed in either round- or flat-bottom microtitration tissue culture plates. Concentrations of responsing and stimulating cells are varied, depening on the experiment. Significant discrimination between isogeneic and allogenic mixtures is possible after 120 hours' culture when cells are labeled ([3H]thymidine) for the last 16 to 18 hours of the test.

Peanut agglutinin as a surface marker for canine T lymphocytes.

Peanut agglutinin (PNA) and surface immunoglobulin (SIg) were investigated as markers for T and B lymphocytes in blood and lymphoid tissues of dogs of various ages. In the blood study, 4 age groups (n = 8 dogs/group) were used. The mean (+/- SD) percentages of PNA-positive (PNA+) cells were 68.4 +/- 8.6% (group 1, less than 1 year old), 70.3 +/- 9.2% (group 2, 1 to 2 years old), 72.0 +/- 3.7% (group 3, 5 to 6 years old), and 63.8 +/- 10.1% (group 4, 10 to 11 years old). The mean percentages of SIg-positive (SIg+) cells in blood were 32.1 +/- 10.6% (group 1), 43.2 +/- 7.0% (group 2), 34.3 +/- 4.8% (group 3), and 35.0 +/- 6.8% (group 4). The mean total percentages of PNA+ and SIg+ cells were 100 +/- 6.0% (group 1), 113.5 +/- 4.9% (group 2), 106.3 +/- 5.3% (group 3), and 98.9 +/- 9.2% (group 4). The proportions of PNA+ and SIg+ cells in dogs of group 2 were significantly (P less than 0.05) different from those in dogs of the other groups. Serial changes in PNA+ and SIg+ cells were investigated in blood of 6- to 29-week-old pups (n = 8). A significant (P less than 0.05) transient decrease in PNA+ cells and a corresponding increase in SIg+ cells was observed in pups between 14 and 17 weeks old. Lymphoid tissue specimens and blood samples were obtained from 2- to 6-month-old dogs (n = 11) and from 6- to 12-month-old dogs (n = 10).(ABSTRACT TRUNCATED AT 250 WORDS)

Efficacy of ivermectin in paste and injectable formulations against microfilariae of Onchocerca cervicalis and resolution of associated dermatitis in horses.

The efficacy of a single dose of ivermectin (0.2 mg/kg), in injectable or paste formulations, against microfilariae of Onchocerca cervicalis and associated skin lesions was evaluated in 20 naturally infected horses during midsummer months in Louisiana. All horses had clinical signs of dermatitis of the ventral midline and/or limbs, shoulders, thorax, and withers. Efficacy was monitored at 21, 42, and 63 days after treatment. Procedures done at these intervals included microfilarial counts of 6-mm skin biopsy specimens of affected ventral midline, grading of gross lesions, and photography and histologic assessment of ventral midline biopsy specimens. Microfilarial numbers were reduced to 0 by 21 days after treatment in all but one horse. Active lesions improved or were resolved completely by 63 days after treatment. Total inflammation, as judged by histologic methods, was reduced in all horses by 63 days after treatment, but there was a residual population of inflammatory cells in all horses. Adverse reactions after treatment were not observed in any of the horses.

Immunodeficiency manifested by oral candidiasis and bacterial septicemia in foals.

Oral candidiasis and bacterial septicemia were diagnosed in 8 foals that had laboratory and/or pathologic evidence of immunodeficiency. Two foals suffered solely from complete failure of passive transfer of colostal immunoglobulins. Six foals had evidence of immune defects but did not meet the criteria for diagnosis of any of the currently recognized primary equine immunodeficiency syndromes. All six of these foals died or were euthanatized due to bacterial infections. One foal with failure of passive transfer recovered and the other died of a mesenteric torsion before the effect of treatment could be evaluated.

Preputial hematoma in a stallion.

An 8-year-old Quarter Horse stallion was admitted with a swelling involving the prepuce, dorsal part of the scrotum, and root of the penis. The swelling was first noticed 10 days after breeding, with no evidence that it had resulted from a kick by the mare. The penis was retracted into the prepuce, except for a 10- to 15-cm protrusion of the glans penis. Treatment included warm hydrotherapy, massage, stockinette compression, antidiuretics, antibiotics, penile support, and exercise, without success. At necropsy, a massive preputial hematoma was found extending forward from the root of the penis to 12 cm proximal to the glans penis. The exact origin of the hematoma could not be determined. Most likely, bleeding had originated from the vascular plexus lying subfascially on the surface of the penis outside the intact tunica albuginea or from a branch of the external pudendal artery.

Chronic chondritis of the equine arytenoid cartilage.

Chronic chondritis of the arytenoid cartilage was diagnosed in 7 male Thoroughbred horses examined for obstructive upper airway disorders. The history of the cases was characterized by a 3- to 6-month progression of exercise intolerance and inspiratory dyspnea during exercise. Endoscopy revealed marked asymmetry of the rima glottidis, partial or complete inability to abduct the involved cartilage, and axial displacement of the involved arytenoid cartilage. In less severe cases, the disorder was confused with laryngeal hemiplegia. Focal elevated lesions of the involved cartilage, which were frequently seen, produced contact lesions on the contralateral cartilage. Subtotal arytenoidectomy was performed in 6 cases. Histologic examination of the removed cartilages revealed marked lamination of the cartilage with fibrous connective tissue. Granulating sinus tracts were seen in 3 cases. The surgical intervention resulted in marked improvement of exercise intolerance in 5 cases.

Ataxia in four horses with equine infectious anemia.

In 4 horses with equine infectious anemia (EIA), the predominant clinical sign was ataxia. Other clinical and laboratory findings often associated with EIA included weight loss, anemia, pyrexia, thrombocytopenia, hemorrhages, hypergammaglobulinemia, and high activity of biliary epithelial enzymes. Neuropathologic findings were nonsuppurative granulomatous ependymitis, meningitis, and encephalomyelitis and plasmacytic-lymphocytic infiltration of the brain and spinal cord. The onset of neurologic signs corresponded to the acute stage of infection in at least 2 horses, and the signs developed at least 18 months after infection in 1 case. Cerebrospinal fluid from 3 of the horses contained high concentration of protein and white cells, although changes in 1 horse may have been associated with a prior traumatic attempt to collect CSF. By comparison, CSF from 3 ponies inapparently infected with EIA was normal. Active production of anti-EIA antibody in the CSF was suspected on the basis of serologic findings.

Neonatal isoerythrolysis in mule foals.

Four mule foals received anti-RBC antibody from their dam's colostrum. Two foals developed anemia, 1 developed severe thrombocytopenia, and 1 did not develop any clinical or laboratory abnormalities. All foals had the same donkey sire. Seven mares, including the dams of foals 1 to 4, bred to the donkey sire produced anti-RBC antibody. Two mares produced anti-RBC antibody to the donkey RBC on the first mating. The donkey sire and 18 of its offspring were found to have an RBC antigen unique to mules and donkeys. Thus, all matings of this donkey to horse mares were incompatible. The reason for the high rate of sensitization of mares bred to this donkey is unknown.

Seminal vesiculitis as a cause of signs of colic in a stallion.

A 5-year-old stallion was referred because of signs of abdominal pain. During the initial examination, signs of pain were elicited when the right seminal vesicle was palpated per rectum. Signs of pain were also elicited during sexual arousal and attempts at semen collection. The right seminal vesicle was subsequently determined to be abnormal by ultrasonographic and endoscopic examination. The stallion was treated with trimethoprim/sulfamethoxazole for 6 weeks. Five months later, there had been no recurrence of the condition.

Complications associated with immunotherapy of equine phycomycosis.

Five horses with pythiosis of the limbs were treated unsuccessfully by surgery or topical application of amphotericin B, or both. Follow-up immunotherapy resulted in 1 horse responding favorably. Three horses were cured of the fungal infection but developed osteitis or deep-seated laminitis, which necessitated their destruction. The remaining horse, which had severe anemia, died before the course of vaccination was completed.

Equine autoimmunity.

Although relatively little is known about autoimmunity and autoimmune mechanisms specifically in horses, the similarities between clinical syndromes with identifiable effector mechanisms in horses and other species suggest that comparable mechanisms may be applicable. Our understanding of equine autoimmunity undoubtedly will benefit from the extensive study of autoimmunity in other species.