Maria Cristina da Silva

Conidiobolomicose em ovinos no Estado de Mato Grosso

The epidemiological, clinical, pathological and mycological findings of an outbreak of conidiobolomycosis in a flock of 40 Santa Ines sheep, in the state of Mato Grosso, Brazil, are reported. The illness occurred in the municipality of Nobres during January-June, 2007, resulting in death of about 30% of the affected sheep within 2-5 weeks. The clinical signs were characterized by apathy, weight loss, labored and noisy breathing with dyspnea, and mucous or serohemorrhagic nasal discharge. In three sheep there was unilateral exophthalmia, keratitis and corneal ulceration. A firm whitish, multi-lobulated, friable growth was evident in the ethmoturbinate region at the coronal sections of the head from six affected sheep. In all sheep the choana had similar nodular infiltration which resulted in local obstruction. In three of those there was exophthalmia. There was infiltration in the cribiform plate and brain (2 cases), regional lymph nodes (2), lungs (3), and abomasums (1). Microscopic findings were granulomatous inflammation of the rhinoetimoidal region, with necrosis, lymphocytic infiltration, epithelioid multinucleated giant cells and fibrovascular tissue, surrounding Splendore-Hoeppli material wich contained unstained ghost images of hyphae. The methenamine-silver stain uncover fungi hyphae, rarely ramified with bulbous dilatation in their extremities. Conidiobolus sp. was isolated from nasal tissue lesions of four sheep.

Pathology of Nasal Infection caused by Conidiobolus lamprauges and Pythium insidiosum in Sheep

Conidiobolomycosis and pythiosis are important diseases of sheep in midwestern Brazil. Veterinary practitioners consider it difficult to differentiate between these diseases because they have similar clinical features. In this study, 186 sheep were subjected to necropsy examination over a 6-year period. Thirty (16.1%) cases of rhinitis in sheep that were caused by Conidiobolus lamprauges (n = 15) or Pythium insidiosum (n = 15) were investigated further. The lesions of C. lamprauges infection were mainly rhinopharyngeal (86.7%), localized to the ethmoidal region and associated with exophthalmos. The lesions appear as a white to yellow, firm mass that microscopically appears as a granulomatous inflammatory reaction with numerous giant cells. In contrast, P. insidiosum infection is associated with rhinofacial (93.3%) lesions that mainly involve the frontal region and hard palate and appear as an irregular, friable, yellow to red mass. Microscopically, pythiosis presents as diffuse necrotizing eosinophilic rhinitis. Immunohistochemistry using polyclonal antisera raised specifically against the two organisms was used to confirm the identity of the infectious agent in each disease. This study reports the first immunohistochemical diagnosis of conidiobolomycosis and the first description of a rhinopharyngeal lesion caused by P. insidiosum in sheep.

Molecular characterization of ovine zygomycosis in central western Brazil.

Zygomycosis is an important granulomatous disease that affects humans and animals, particularly sheep in tropical regions. Rhinofacial and nasopharyngeal zygomycosis were described in sheep in association with Conidiobolus spp. The present study characterized 5 samples of Conidiobolus isolated from 3 herds with clinical disease in Mato Grosso State, Brazil. The clinical and pathological findings were similar to nasopharyngeal zygomycosis. Based on morphological features, isolates were classified as Conidiobolus spp., and molecular phylogenetic analyses based on 18S ribosomal DNA grouped all isolates in a Conidiobolus lamprauges cluster. The current report describes the molecular characterization of ovine nasopharyngeal zygomycosis associated with C. lamprauges.

Prevalência de Salmonella sp. em suínos abatidos no Estado de Mato Grosso

The present study aims to determine the prevalence of Salmonella sp. in pigs in slaughterhouses under Federal Inspection at Mato Grosso, Brazil. Samples of mesenteric lymph nodes and tonsils were collected in three abattoirs from different regions. Three hundred samples were processed; fifty (16.6%) were positive for Salmonella sp. and belonged to fourteen different serological types. The most common serotypes were Derby (16%), Typhimurium (14%), London (12%) and Give (12%). The epidemiological results obtained show a low prevalence of this bacterium when compared to studies in other Brazilian states.

Development and application of polymerase chain reaction test for detection of Conidiobolus lamprauges

ABSTRACT.- Silveira M.M., Paula D.A.J., Silva M.C., Pitchenin L.C., Cruz R.A.S., Colodel E.M., Dutra V. & Nakazato L. 2013. Development and application of polymerase chain reaction test for detection of Conidiobolus lamprauges . Pesquisa Veterinaria Brasileira 33(12):1448-1452. Departamento de Clinica Medica Veterinaria, Faculdade de Agronomia, Medicina Ve-terinaria e Zootecnia, Universidade Federal de Mato Grosso, Av. Fernando Correa da Costa 2673, Bairro Boa Esperanca, Cuiaba, MT 78068-900, Brazil. E-mail: lucnak@ufmt.brConidiobolomycosis is a granulomatous disease caused by the fungus Conidiobolus spp. in humans and animals. Traditional technique for diagnosis of the disease is isolation of the agent associated with the presence of typical clinical signs and pathological conditions. The aim of this study was to describe the development of a specific polymerase chain reaction (PCR) test for Conidiobolus lamprauges to detect the fungus in clinical samples. Samples from suspected animals were collected and submitted to isolation, histopathological analy-sis and amplification by PCR. DNA from tissues was subjected to PCR with fungi universal primers 18S rDNA gene, and specific primers were designed based on the same gene in

Genes termorregulados diferencialmente expressos em Conidiobolus lamprauges

Conidiobolus lamprauges is a pathogen zygomycetes fungi of humans and animals, responsible for conidiobolomycosis, which is characterized by a severe granulomatous chronic rhinosinusitis. The ability to adapt and grow at high temperatures is suggested as an attribute of virulence in fungi that infect animals and humans, however regarding C. lamprauges little information is available about this aspect. This paper aims to identify differential expression genes in C. lamprauges grown at 30°C and 37°C through the technique of Representational Difference Analysis (RDA). After the analysis and sequencing of a set of 120cDNAs, it was identified enolase, a glycolytic enzyme, differentially expressed at 37°C. This gene performs functions related to pathogenicity during host-pathogen interaction process in several pathogenic microorganisms. showing a potential involvement in host-pathogen relationship, and virulence in C. lamprauges.

Prevalência de Streptococcus suis tipo 2 por meio da técnica de reação em cadeia da polimerase em suínos abatidos no Estado do Mato Grosso

Streptococcus suis is a pathogen that affects the industrial production of swine worldwide. It is extremely important, because it is associated with pigs and humans diseases. The aim of this study was to determine the prevalence of Streptococcus suis type 2 in 201 samples of tonsils from clinically healthy animals by the PCR technique. The samples positive for S. suis type 2 were tested for the gene encoding extracellular factors (ef). The results showed that the prevalence (23.38%) was higher than other recent survey in the State, demonstrating that the PCR is a more sensitive method in relation to the bacterial isolation. There was a low occurrence of ef* gene in samples (1.49%) showing great importance to local swine population, because negative strains are potentially less virulent that positive strains.

Preliminary virulence genotyping and phylogeny of Escherichia coli from the gut of pigs at slaughtering stage in Brazil

In the current study we screened Escherichia coli from intestine of pigs slaughtered in Mato Grosso, Brazil, for virulence-markers related to human disease. Furthermore, we employed for the first time a phylogenetic assay to explore the association between phylogeny and virulence genotype in E. coli from finished swine. A low prevalence (7.8%) of E. coli harbouring virulence genes was observed. Among the positive isolates, 3.3% could be classified as atypical EPEC, 2.2% as STEC and 2.2% as CDT harbouring E. coli. Virulence genes were not found to co-occur in a strain. Phylogenetic determination of isolates revealed a low prevalence of E. coli lineages related to disease. Therefore, preliminary sampling of 74 pigs indicated that slaughter swine may not be major reservoirs of E. coli capable of causing human disease. In light of the significant association between phylogeny and virulence genotype, we also underscored the phylogenetic grouping of strains as a valuable tool for E. coli surveillance programmes in slaughterhouses.

Prognostic significance of Ki67 and its correlation with mitotic index in dogs with diffuse large B-cell lymphoma treated with 19-week CHOP-based protocol:

Diffuse large B-cell lymphoma (DLBCL) is the most common type of lymphoma in dogs. We evaluated Ki67 immunoexpression and mitotic index (MI) in dogs diagnosed with DLBCL and treated with a 19-wk CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) protocol. Twenty-nine lymph node samples from dogs diagnosed with DLBCL were analyzed for Ki67 immunostaining, and positive cells present in 1 cm2 were counted in a grid reticle for comparison of survival times above and below the means. The Ki67 mean was 107, and the MI mean was 21. There was a significant (p < 0.05) difference in median survival time between Ki67 immunostaining above and below the mean, with no difference in MI groups. Ki67 values >107 positive cells per 5 HPF counted in a grid reticle were associated with shorter survival times in dogs with DLBCL treated with a 19-wk CHOP-based protocol.

Proteínas imunorreativas de Conidiobolus lamprauges isoladas de ovinos infectados naturalmente

The study of sheep conidiobolomycosis has been carried out in its clinical, epidemiological, pathological and molecular aspects. Information, however, about the host immune response in infection Conidiobolus lamprauges is absent. This study aimed to identify immunoreactive proteins that may play an important role in the immune response of sheep naturally infected by C.lamprauges. For protein and immunological characterization, C. lamprauges (strain FIOCRUZ-INCQS 40316) isolated from a sheep with clinical signs of conidiobolomycosis in the MT state and five sera samples of naturally infected sheep were used. The presence of IgG antibody was observed in all patients with reagent titers in dilutions up to 1:1600. In immunoblot technique, the antigenic profile against infected sheep sera showed twelve reactive bands with molecular weights ranging from 35 to 198 kDa. Among them, the 198 kDa protein was reactive against sera from three sheep and the 53 kDa showed increased intensity compared to other bands probably being immunodominant. Healthy animal serum samples showed no reactivity demonstrating the specificity of the technique. The presence of antigenic proteins of C. lamprauges and specific IgG in sheep sera observed in this study may assist in the development of early diagnostic methods and the use of protein as candidate vaccines for the control and prevention of infection in animals and human.