Immunisation with Neospora caninum subunits rsNcSAG4 and rsNcGRA1 (NcSAG4 and NcGRA1 epitopes construct) in BALB/c mice: the profile of the immune response and controlling the vertical transmission.

Neospora caninum is an apicomplexan protozoan that causes neosporosis, which has a high economic impact on cattle herds with no available vaccine. During infection, the secretion of dense granules and the expression of surface antigens play an important role in hosting immunomodulation. However, some epitopes of those antigens are immunogenic, and using these fractions could improve the subunit antigens in vaccine design. This study evaluates the recombinant peptides rsNcGRA1 and rsNcSAG4 derived from NcGRA1 and NcSAG4 native antigens as vaccine candidates produced by a fermentative process in the yeast culture system of Komagataella phaffii strain Km71, confirmed by colony PCR, SDS-PAGE, and western blotting. The assay was conducted in BALB/c mice using the peptides at low (25 µg) and standard (50 µg) dosages in monovalent and combined administrations at three time points with saponin as an adjuvant assessing the immunogenicity by antibodies response and cytokine production. We challenge the females after pregnancy confirmation using 2 × 105 NC-1 tachyzoites previously propagated in Vero cells. We assessed the chronic infection in dams and vertical transmission in the offspring by PCR and histopathology. Mice, especially those immunised with combined peptides and monovalent rsNcGRA1 at a standard dose, controlling the chronic infection in dams with the absence of clinical manifestations, showed an immune response with induction of IgG1, a proper balance between Th1/Th2 cytokines and reduced vertical transmission in the pups. In contrast, dams inoculated with a placebo vaccine showed clinical signs, low-scored brain lesions, augmented chronic infection with 80% positivity, 31% mortality in pups, and 81% vertical transmission. These findings indicate that rsNcGRA1 peptides in monovalent and combined with rsNCSAG4 at standard dose are potential vaccine candidates and improve the protective immune response against neosporosis in mice.

Evaluación clínica e histopatológica de la pitiosis cutánea en burros (Equus asinus) / Clinical and Histopathological Evaluation of Cutaneous Pythiosis in Donkeys (Equusasinus) / Avaliação clínica e histopatológica da pitiose cutânea em burros (Equus asinus)

El objetivo de este estudio fue evaluar y caracterizar los aspectos clínicos e histopatológicos de la pitiosis cutánea en burros (Equus asinus) en el departamento de Córdoba, Colombia. Se realizó un estudio de tipo descriptivo, no probabilístico en animales de conveniencia. Fueron analizados nueve burros con pitiosis cutánea diagnosticados clínica e histopatológicamente. Al describir los casos se observó una grave ulceración granulomatosa en forma de cráter, con tejido necrótico, apariencia tumoral, abundante exudación filante fibrino-sanguinolenta y salida de material caseificado de color blanco amarillento denominado kunkers. Las lesiones se ubicaron en miembros, abdomen ventral y pecho de los animales. Histopatológicamente, con la coloración de hematoxilina-eosina, se observó dermatitis piogranulomatosa severa, con abundante infiltrado inflamatorio de tipo eosinofílico y neutrofílico; con la coloración de Grocott se evidenciaron hifas con tabiques y parcialmente ramificadas en ángulo recto, de color café. El diagnóstico definitivo de la enfermedad se fundamentó en las características clínicas, el diagnóstico diferencial y en los hallazgos histopatológicos. Fue concluyente el diagnóstico de la pitiosis cutánea. El presente estudio se convierte en el primer reporte de esta enfermedad en burros (Equus asinus) en el departamento de Córdoba y en Colombia.

The purpose of this study was to evaluate and characterize the clinical and histopathological aspects of cutaneous pythiosis in donkeys (Equus asinus) in the Department of Cordoba, Colombia. A descriptive, non-probability study was conducted on domesticated animals. Nine donkeys clinically and histopathologically diagnosed with cutaneous pythiosis were analyzed. After describing the cases, a serious, crater-shaped granulomatous ulcer was observed, with necrotic tissue, a tumor appearance, abundant fibrinous-bloody exudation and yellowish-white caseous material known as kunkers. The lesions were found in the members, ventral abdomen and chest of the animals. Histopathologically, with the hematoxylin-eosin staining, severe pyogranulomatous dermatitis was observed, with abundant eosinophilic and neutrophilic inflammatory infiltrate; with the Grocott staining, hyphae with septa were found, partially branching into a right angle and brown in color. The definitive diagnosis of the disease was based on the clinical features, the differential diagnosis and the histopathological findings. The diagnosis of cutaneous pythiosis was conclusive. This study becomes the first report of this disease in donkeys (Equus asinus) in the department of Córdoba and in Colombia.

O objetivo deste estudo foi avaliar e caracterizar os aspectos clínicos e histopatológicos da pitiose cutânea em burros (Equus asinus) no Estado de Córdoba, Colômbia. Realizou-se um estudo de tipo descritivo, não probabilístico em animais de conveniência. Foram analisados nove burros com pitiose cutânea, diagnosticados clínica e histopatologicamente. Ao descrever os casos se observou uma grave ulceração granulomatose em forma de cratera com tecido necrótico, aparência tumoral, abundante exsudação pegajosa fibrinosa sanguinolenta e saída de material caseoso de cor branca amarelada denominada kunkers. As lesões se localizaram em membros, abdômen ventral e peito dos animais. Histopatologicamente, com a coloração de hematoxilina-eosina, se observou dermatite pio granulomatosa severa, com abundante infiltrado inflamatório de tipo eosinofílico e neutrofílico; com a coloração de Grocott se evidenciaram hifas com tabiques e parcialmente ramificadas em ângulo reto, de cor marrom. O diagnóstico definitivo da doença se fundamentou nas características clínicas, o diagnóstico diferencial e nos achados histopatológicos. Foi concluinte o diagnóstico da pitiose cutânea. O presente estudo se converte no primeiro relatório desta doença em burros (Equus asinus) no Estado de Córdoba na Colômbia.