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.

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.

Immunodiagnostic assays.

The immune system is a complex interactive network. Defects in its function can be characterized broadly as being the result of actual deficiencies in the network or misdirection of normal immunologic functions. The assays that are available to detect deficiencies in the immunologic network barely scrape the surface of the possibilities. These assays primarily evaluate humoral immune function, but undetected defects in innate and cellular immunity are sure to exist. Although assays of humoral immunity have allowed the characterization of a number of immunodeficiency syndromes in horses, closer evaluation of the equine immune system using newer assays described in this text, as well as future assays yet to be developed are sure to determine new syndromes. Assays of misdirected immunologic functions have been limited to detection of misdirected antibody responses, but the dependence of antibody production on help from T cells could reflect an underlying defect of cellular immunity. Similar to immunodeficiency syndromes, misdirected responses of the innate and adaptive arms of immunity are sure to occur but will only be detected by more diligent surveillance of diseased horses and application of new immunodiagnostic technologies.

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.

Characterization of a red blood cell antigen in donkeys and mules associated with neonatal isoerythrolysis.

A red cell antigen of donkeys and mules was identified using antibodies in serum from a mare which produced a mule foal affected with neonatal isoerythrolysis (NI). Subsequently antibodies with similar activity were identified in the sera of other mares which had produced mule foals and were produced by immunization of horses with blood from donkeys. The antigen detected by these antibodies does not correspond to any recognized horse red cell alloantigen. This may be a xenoantigen since all donkeys (and mules) tested have shared this antigen and all horses tested have lacked the antigen. The results suggest that all mule pregnancies (donkey sire x horse dam) are incompatible with regard to this factor and a potential for neonatal isoerythrolysis exists in all cases.

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.

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)

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.

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.

Equine disease association studies: a clinician's perspective.

Diagnostic criteria should be carefully defined and described in disease association studies to allow (1) comparison among studies from different laboratories evaluating the same disease, (2) critical evaluation of selection procedures of patients, and (3) to strengthen genuine associations with any genetic marker system. Factors to consider include age at onset of disease, specialized diagnostic methods necessary to diagnose or eliminate patients with a selected disease, ranges of affectedness and differences in sex expression.

Association of arytenoid chondritis with equine lymphocyte antigens but no association with laryngeal hemiplegia, umbilical hernias and cryptorchidism.

Associations were sought between ELA A1-A10 and W11 antigens and the presence of laryngeal hemiplegia, arytenoid chondritis, umbilical hernias and cryptorchidism in Thoroughbreds and/or Quarter Horses. No significant associations were detected between laryngeal hemiplegia and any ELA antigen in Thoroughbreds. The association between arytenoid chondritis and A9 was significant with a relative risk (RR) of 15.6 and aetiologic fraction (EF) of 0.80 in Thoroughbreds. There were apparent associations based on RR between A4 and A5 in Quarter Horses with umbilical hernias (RR = 7.5 and 6.1 respectively); however, these were not statistically significant. No significant associations were detected with cryptorchidism in Quarter Horses when the control population included both sexes. When only unaffected males were used as the control group, there was an apparent increase in relative risk with A6 (from RR = 1.7 to 4.3); however this was not statistically significant. Cryptorchidism in Thoroughbreds showed an increased relative risk with A5 regardless of whether the control population included males and females (RR = 4.1) or only males (RR = 4.7) but the increases were not statistically significant.

Joint Report of the Fifth International Workshop on Lymphocyte Alloantigens of the Horse, Baton Rouge, Louisiana, 31 October-1 November 1987.

Six laboratories participated in the Fifth International Workshop on Lymphocyte Alloantigens of the Horse, testing 132 alloantisera against lymphocytes of 880 horses chosen to represent different families and breeds. Most of the alloantisera were produced by lymphocyte immunization between horses matched at the ELA-A locus. All horses were also tested with antisera contributed to the workshop by participating laboratories which identified ELA specificities A1-A10 and W12-W21. Previously identified workshop specificities ELA-W14, W15 and W19 were accepted as products of the ELA-A locus based on family and population studies by the workshop. Their designations were changed to ELA-A14, ELA-A15 and ELA-A19, respectively. Two new specificities were identified, namely ELA-W22 (W22) and ELA-W23 (W23). Population and family studies indicated that W22 and W23 as well as W13 are products of an ELA locus other than ELA-A. The presence of these specificities was correlated with the presence of certain ELA-A locus specificities, e.g. W13 with A3, W22 with A2 and W23 with A5. However, the association was not complete and W13, W22 and W23 also segregated with other ELA-A specificities in some families. Evidence for recombination was found between the ELA-A locus and the locus or loci encoding these specificities resulting in seven recombinant haplotypes found among the data presented in this workshop. Further studies are required for definitive assignment of the specificities to a class I or class II locus.

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.

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.

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).