The Rhipicephalus microplus tick is a notorious blood-feeding ectoparasite of livestock, especially cattle, responsible for massive losses in animal production. It is the main vector for transmission of pathogenic bacteria and parasites, including Babesia bovis, an intraerythrocytic apicomplexan protozoan parasite responsible for bovine Babesiosis. This study describes the development and testing of a live B. bovis vaccine expressing the protective tick antigen glutathione-S-transferase from Haemaphysalis longicornis (HlGST). The B. bovis S74-T3B parasites were electroporated with a plasmid containing the bidirectional Ef-1α (elongation factor 1 alpha) promoter of B. bovis controlling expression of two independent genes, the selectable marker GFP-BSD (green fluorescent protein–blasticidin deaminase), and HlGST fused to the MSA-1 (merozoite surface antigen 1) signal peptide from B. bovis. Electroporation followed by blasticidin selection resulted in the emergence of a mixed B. bovis transfected line (termed HlGST) in in vitro cultures, containing parasites with distinct patterns of insertion of both exogenous genes, either in or outside the Ef-1α locus. A B. bovis clonal line termed HlGST-Cln expressing intracellular GFP and HlGST in the surface of merozoites was then derived from the mixed parasite line HlGST using a fluorescent activated cell sorter. Two independent calf immunization trials were performed via intravenous inoculation of the HlGST-Cln and a previously described control consisting of an irrelevant transfected clonal line of B. bovis designated GFP-Cln. The control GFP-Cln line contains a copy of the GFP-BSD gene inserted into the Ef-1α locus of B. bovis in an identical fashion as the HIGST-Cln parasites. All animals inoculated with the HlGST-Cln and GFP-Cln transfected parasites developed mild babesiosis. Tick egg fertility and fully engorged female tick weight was reduced significantly in R. microplus feeding on HlGST-Cln-immunized calves. Collectively, these data show the efficacy of a transfected HlGST-Cln B. bovis parasite to induce detectable anti-glutathione-S-transferase antibodies and a reduction in tick size and fecundity of R. microplus feeding in experimentally inoculated animals.

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Pesq. Vet. Bras. 34(2):141-146, fevereiro 2014 141 RESUMO.-Este trabalho foi desenvolvido com o objetivo de avaliar a resistência aos ectoparasitas em bovinos jovens da raça Crioula Lageana e meio-sangue Angus em condições naturais. Foram utilizados 10 machos castrados em cada grupo, recém desmamados (6-8 meses) e mantidos sob as mesmas condições de manejo, em pastagens cultivadas de inverno e verão, no município de Monte Castelo -SC, sofrendo infestação natural por ectoparasitas. A cada 28 dias, fêmeas de carrapatos (Rhipicephalus (Boophilus) microplus) com tamanho acima de 4 mm foram quantificadas nos dois lados do corpo de cada animal, bem como os nódulos com larvas de Dermatobia hominis. A espessura da capa do pelame foi também avaliada a cada coleta e os animais foram classificados quanto à coloração do pelame. Os animais da raça Crioula Lageana foram mais resistentes às infestações por D. hominis e R. microplus. Não houve diferença entre a disposição de carrapatos e bernes conforme os lados direito e esquerdo dos animais. Os animais de pelagem escura albergaram a maior porcentagem de ectoparasitas nos grupos avaliados. This study was carried out to evaluate the breed resistance against ectoparasites infestations in Crioula Lageana and crossbred Angus male calves under natural condition. Ten weaned and neutered male calves (6-8 months) of each group were kept together in winter and summer pastures in Monte Castelo, Santa Catarina State. Every 28 days, female ticks (Rhipicephalus (Boophilus) microplus) more than 4 mm long were counted on the right and left side of the body of each animal, as well as the D. hominis nodules. Coat thickness also was measured at each sampling and all animals were classified according coat color. Crioula Lageana group was less infested by D. hominis than crossbred Angus in five times and by ticks in the infestation peak (P<0.05). There were no differences in the distribution of the ticks and grubs between animal body sides. Dark-haired animal in both groups were the most susceptible.

Neuroblastic tumors can originate from the central neuraxis, olfactory epithelium, adrenal medullary region or autonomous system. Ganglioneuroblastoma are a type of neuroblastic tumor, with very few case descriptions in animals. Diagnosis of facial nerve ganglioneuroblastoma was made in a feline leukemia virus-positive 11-month-old cat. The cat had hyporexia, left head tilt, depressed mental state, horizontal nystagmus, inability to retract the pinched left lip, anisocoria, ptosis, and absence of the menace reflex. Gross necropsy showed a mass at the left facial nerve root region. Histological examination of this mass showed neoplastic proliferation of neuroblasts arranged in a cohesive pattern and mature ganglion cells. Ganglion cells were positive for neurofilament, neuron-specific enolase, S100, and glial fibrillary acidic protein by immunohistochemistry, while neuroblasts were positive for vimentin, S100, neuron-specific enolase and feline leukemia virus.

Background: Leishmaniasis is a chronic infectious disease caused by intracellular protozoan Leishmania that affects canine and human. The visceral leishmaniasis (VL) is caused by the Leishmania donovani complex, in which canines are the main reservoir for human VL. In Southern Brazil, case reports of this disease have increased, especially when canines are infected in endemic areas in the country. Canines usually present a systemic disease, characterized by cutaneous lesions, weight loss, generalized lymphadenopathy, hepatomegaly and splenomegaly. This report aims to describe the clinicalpathological features of a case of autochthonous VL in a canine of an urban area of Porto Alegre.Case: A male, mixed breed, dog , 7 year-old, resident at the East Side of Porto Alegre, living together with two other canines, presented at clinical examination progressive weight loss, associated with hyporexia and hyperthermia, evolving to lateral recumbency, paralisys, and death. At necropsy, the dog was cachectic with diffusely pale mucosae. Gross findings included liver enlarged, with multifocal firm brownish areas, spleen enlarged, showing multifocal firm round dark-red areas, and kidneys diffusely pale with evidentiated tranversal striations on cortical zone. At microscopic examination, there was on the spleen diffuse inflammatory infiltrate of macrophages with large cytoplasm containing multiple amastigotes. The liver, exhibited atrophy of hepatocytes and moderate multifocal inflammatory infiltrate in sinusoids of macrophages containing multiple amastigotes. These features were also observed moderately on lymph nodes and lamina propria of large intestine. Immunohistochemistry examination showed marked positive staining for Leishmania spp. in amastigotes located whitin the cytoplasm of macrophages of spleen, liver, lymph nodes and large intestine.Discussion: Canine leishmaniasis is a disease that affects both internal organs and skin. However, the condition is called VL mostly because it is associated with the same species that cause the visceral form in humans. In this case the canine presented liver, spleen and lymph node lesions; however no skin lesion was observed during the clinical examination. A serological evaluation of canids in East Side of Porto Alegre performed in 2005 showed that 3.5% of these dogs were positive for Leishmania; yet neither of these animals presented clinical signs. This demonstrates that the agent was already present, however only 3-10% of the canines infected develop clinical disease. In non-endemic areas VL is related to the migration of canids from endemic areas, where VL is common. However the canine here described was born, raised, and was living in Porto Alegre. Lutzomyia longipalpis is the main vector of VL and despite its wide distribution it was identified only in 2009 in Rio Grande do Sul State. Autochthonous cases of VL were reported in canines and humans in this state, confirming the hypothesis that the vector was widely distributed and it is now present in Porto Alegre, keeping the epidemiological cycle of that disease active. Canine VL occurs prior in humans, since canines are the main domestic reservoirs and are critical for the maintenance of this disease cycle. This is the first autochthonous VL in a canine of an urban area of Porto Alegre; therefore authorities should be alert, and new control measures must be taken to avoid the canine leishmaniasis outbreak due to its potential for zoonotic transmission.Keywords: leishmaniasis, canine, protozoan, immunohistochemistry.