Immunological and pathological investigations in equine experimental uveitis

Background

The pathogenic mechanism of equine recurrent uveitis (ERU) is still poorly defined and many variations between experimental animal models and spontaneous disease exist.

Objectives

The aim of our study was to investigate if Th17 cell-mediated response plays role in the pathogenesis of the used experimental model in horses and to reveal its pathological findings.

Methods

Experimental uveitis was induced in 6 healthy horses. The concentrations of retinal autoantigen CRALBP and IL-17 were measured using ELISA in aqueous humor and vitreous body of the 12 inflamed eyes as well as in 12 control non-inflamed eyes taken from 6 horses in slaughter house. After centrifugation of the two eye media, smears were prepared and cytological investigation was performed. Tissue specimens were taken from all eye globes and were submitted to histopathological investigation.

Results

CRALBP and IL-17 concentrations were significantly elevated in eye media of horses with experimental uveitis in comparison with controls. Cytological and histopathological findings corresponded to the changes characteristic of chronic immune-mediated inflammation with mononuclear cell infiltration of uvea, choroid, retina, and eye media as well as severe retinal destruction.

Conclusions

Our study demonstrated the involvement of the retinal autoantigen CRALBP as well as IL-17 in the pathogenesis of experimental uveitis in horses. These findings suggests that this experimental uveitis in horses may serve as a suitable animal model for investigation of IL-17- mediated immune response during spontaneous autoimmune uveitis in horses as well as in humans.

     

Use of Non‐invasive Methods for Evaluating the Testicular Biometry in Collared Peccaries (Pecari tajacu Linnaeus, 1758)

The aim of this study was to compare the accuracy of two methods used to estimate testicular volume in the collared peccary. Calliper and ultrasonographic measurements of testicular dimensions (length, width and height) of both testes were taken on five adult collared peccaries. The testicular volume was calculated by Lambert's empiric formula: length (L) × width (W) × height (H) × 0.71, the formula of an ellipsoid L × W × H × 0.52, and Hansen's formula: L × W² × 0.52. The calculated volumes were then compared with the actual ones, which were estimated by water displacement. The mean of true testicular volume was 22.65 ± 1.52 ml. Lambert's formula estimated testicular volume more accurately when ultrasound measurements were taken. However, when the calliper was the methodology used, the results were closest to the true volume, especially when Ellipsoid formula and Hansen's formula were applied, and underestimated the true volumes by 1.53 ± 1.75 ml and 1.53 ± 1.65 ml, respectively. This specific application of technologies in wild animals has the potential to revolutionize the selection process for the collared peccary entering artificial insemination or natural breeding programmes.     

Isolation of Leptospira noguchii from sheep

The main goal of this study was to obtain new isolates of Leptospira spp. from sheep. A total of 10 kidney samples and 44 blood samples were collected from sheep slaughtered in Pelotas, Southern Brazil. One isolate was obtained which was identified by 16S rRNA gene sequencing and serogrouping to be Leptospira noguchii serogroup Autumnalis. Microscopic agglutination test (MAT) evaluation revealed that 4.5% of the sheep sera reacted against the Autumnalis serogroup. This is the first report of isolation of L. noguchii from sheep. Together these findings indicate that L. noguchii infections may be a potentially important veterinary problem in this domestic animal species.     

Endometrial nitric oxide production and nitric oxide synthases in the equine endometrium: Relationship with microvascular density during the estrous cycle

Nitric oxide (NO) plays an important role in angiogenesis and in the regulation of the blood flow. This study was carried out to investigate (i) the effects of endogenous estrogens and progestins and exogenous progesterone (P₄) (5 ng/ml or 1 μg/ml) or estradiol 17β (E₂β) (50 pg/ml or 1 μg/ml) on in vitro endometrial NO synthesis; (ii) the presence of different isoforms of NO synthase; (iii) and their relationship to microvascular density in the equine endometrium during the estrous cycle. NOS expression was also evaluated in the myometrium. Expression of endothelial and inducible forms of NOS in the uterus was assessed by Western blot and immunocytochemistry. Vascular density in endometrial tissue was determined on histologic sections. In the luteal phase, compared to the follicular phase, endometrial NO production increased without exogenous hormones and with exogenous E₂β (1 μg/ml). Although immunocytochemistry revealed iNOS and eNOS expression in the endometrium, no positive signal for iNOS was detected by Western blot. Endothelial NOS was observed in endometrial glands, endothelial cells, fibroblasts, blood and lymphatic vessels. Endometrial eNOS expression was the highest in the follicular and mid-luteal phases while it was found to be the lowest in the early luteal phase. In the follicular phase, hyperplasia of endometrial tissue with respect to myometrium was detected. No difference in vascular density was present between phases. All together, NO may play some roles in both proliferative and secretory phases of endometrial development in the mare.     

A monoclonal antibody against a canine CD45 homologue: analysis of tissue distribution, biochemical properties and in vitro immunological activity

This report describes the characterisation of a monoclonal antibody (mAb), AB6, which recognises specifically a cluster of canine leukocyte surface molecules. The immunogen used for obtaining the AB6 mAb was a lysate of canine peripheral blood mononuclear cells (PBMC). This novel mAb belongs to the IgG2a isotype, and reacted in Western blot with four different canine leukocyte glycoproteins with apparent molecular weights of 180, 190, 205 and 220 kDa. The AB6 mAb recognised the majority of canine peripheral blood leukocytes as determined by flow cytometry (97%). It also exhibited a broad reactivity pattern against lymphoid and myeloid cells, inhibited the proliferation of mitogen-stimulated canine PBMC and did not recognise human PBMC and murine splenocytes. The biochemical properties, cell and tissue specificity, and in vitro biological activity of the AB6 mAb indicate that it recognises a canine CD45 homologue. The mAb could become a valuable diagnostic and research tool for the evaluation of immune functions in dogs.     

Phylogenetic characterization of rabies virus isolates from Carnivora in Brazil

The incidence of canine rabies has been widely reported in Brazil, and new rabies virus (RV) variants, genetically similar to canine RV, have recently been isolated from foxes. In order to derive the epidemiological characteristics of Brazilian Carnivora RV, Brazilian RVs isolated from dogs, cats, and foxes were genetically analyzed. Brazilian Carnivora RV isolates were divided into 2 main lineages. The predominant lineage was found in dogs and cats, which included the Argentinean and Bolivian Carnivora RV isolates, and was extensively distributed throughout Brazil and surrounding countries. The other lineage consisted of three sublineages containing Brazilian dog and fox RV isolates, with the dog sublineages located on an internal branch of 2 fox sublineages, suggesting that RV transmission events might have occurred between foxes and dogs in the past. These results suggest that contact between dogs and wildlife has the potential to generate new rabies variants and that it is important to control RV infection cycles in both dogs and wildlife to prevent spread of rabies infection.     

Risk factors associated with sero-positivity to Toxoplasma gondii in captive neotropical felids from Brazil

From September 1995 to February 2001, blood samples were collected from 865 neotropical felids belonging to 8 different species. These animals were housed in 86 institutions located in 78 cities of 20 Brazilian states. Our goal was to identify the risk factors associated with sero-positivity to Toxoplasma gondii in captive neotropical felids from Brazil. All serum samples were tested by the modified agglutination test (MAT), using formalin-fixed whole tachyzoites and mercaptoethanol. For each animal an individual questionnaire was filled with questions about tattoo number, felid species, age, sex, origin, number of animals in the group, introduction of new animals in the group, time in the institution, eating meat previously frozen for a period <7 days in the last 6 months, eating meat of run-over or euthanized animals in the last 6 months, predation of rodents or birds in the last 6 months and presence of domestic cats near the enclosures in the last 6 months. The total sero-prevalence was 55% (95% CI: 52%, 57%). We estimated a prevalence of 46% (95% CI: 40%, 54%) for jaguarundi (Puma yagouaroundi); 58% (95% CI: 53%, 63%) for ocelot (Leopardus pardalis); 50% (95% CI: 45%, 56%) for oncilla (L. tigrinus); 54% (95% CI: 46%, 62%) for margay (L. wiedii); 12% (95% CI: 4%, 31%) for Pampas-cat (L. colocolo); 83% (95% CI: 65%, 93%) for Geoffroy's-cat (L. geoffroyi); 64% (95% CI: 50%, 68%) for jaguar (Panthera onca) and 48% (95% CI: 42%, 54%) for puma (Puma concolor). Multiple logistic regression was used to evaluate the association between the variables in the questionnaire and sero-positivity to T. gondii. We concluded that the independent risk factors for toxoplasmosis were: age >3 years (OR = 4.75 [2.75; 8.2]), eating meat previously frozen for a period <7 days (OR = 2.23 [1.24; 4.01]), and consumption of animals that were run-over or euthanized (OR = 1.64; [1.14; 2.37]).