RESUMO

A significant increase in cutaneous leishmaniasis (CL) and its geographic expansion has motivated the development of techniques to help with diagnosis of the disease. Here we describe immunocytochemical (ICC) and immunohistochemical (IHC) techniques for the diagnosis of CL in the laboratory. Polyclonal antibodies and a modified avidin-biotin complex (Ultra Streptavidin(®)) for Leishmania (V.) braziliensis or Leishmania (L.) amazonensis were developed for the present study. In vitro culture and histological sections from experimentally infected tissues were submitted to ICC/IHC techniques. The polyclonal antibody specificity, stability and immunostaining were evaluated. The polyclonal antibodies purified by chromatography (Sephadex(®)) and obtained from L. (V.) braziliensis and L. (L.) amazonensis insoluble antigens presented 83.3% sensitivity, when the presence of antigens was evaluated, i.e., higher than histopathology or any equivalent method (in vitro culture). The polyclonal antibody presented 100% specificity when used against species frequently found in CL lesions. The ICC/IHC techniques developed in the current study were able to recognize amastigotes and antigens from in vivo and in vitro cultures and from biopsies, offering additional help in the diagnosis of CL. This methodology could be beneficially adopted in public health laboratories.

Assuntos

RESUMO

Occurrence of Acanthamoeba in the hospital environment may represent a health risk for patients, since these organisms can cause severe opportunistic illness, such as keratitis, and also can harbor pathogenic agents. We analyzed the dust from some environments of a public hospital in Curitiba, Parana State, Brazil. Two distinct populations of Acanthamoeba were isolated in five locations and morphologically classified as group I and group II according to Pussard and Pons. Isolates were identified as Acanthamoeba by PCR using primers to amplify a region of 18S rDNA, which showed variation in the product length among the isolates. A cloned culture of group II showed greater growth at 37 degrees C and in media with 0.1, 0.5, and 1.0 M mannitol, which are the physiological characteristics of pathogenic Acanthamoeba. Monitoring the presence of Acanthamoeba in hospital units, as well as evaluating the pathogenicity of the isolates, can be an approach to alert the health professionals to improve the disinfection procedures and minimize the risks of treating this problematic disease caused by this protozoan.

Assuntos

RESUMO

This study evaluated occurrence of the Toxoplasma gondii infection in ovine from urban and periurban areas of Curitiba, State of Paraná, Southern of Brazil. Blood samples from 167 sheep from three farms were collected. ELISA assay was used to detect IgG anti-Toxoplasma gondii antibodies. Of 167 ovine samples, 43 (25.75%) were positive. The parasite is widespread among sheep from urban and periurban areas of Curitiba.

Assuntos

RESUMO

Com o objetivo de avaliar a ocorrência da infecção por Toxoplasma gondii, em ovinos das regiões periurbana e urbana do município de Curitiba, Paraná, Região Sul do Brasil, foram coletadas amostras de sangue de 167 ovinos, de três rebanhos. O teste de ELISA foi utilizado para detectar anticorpos anti-Toxoplasma gondii. Dos 167 soros ovinos, 43 (25,75%) apresentaram anticorpos anti-T. gondii. O parasito está difundido entre os ovinos das duas regiões estudadas.

This study evaluated occurrence of the Toxoplasma gondii infection in ovine from urban and periurban areas of Curitiba, State of Paraná, Southern of Brazil. Blood samples from 167 sheep from three farms were collected. ELISA assay was used to detect IgG anti-Toxoplasma gondii antibodies. Of 167 ovine samples, 43 (25.75%) were positive. The parasite is widespread among sheep from urban and periurban areas of Curitiba.

Assuntos

RESUMO

The south region is the only area in Brazil that does not present autochthonous cases of visceral leishmaniasis (VL), however, in the state of Paraná, dogs and humans have been found showing a VL compatible clinical profile. In view of this problem, the present work aimed at isolating and identifying the parasite and determining the cases autochthony. All animals clinically suspect of VL were clinically evaluated, and had samples of their blood collected for hemoculture (NNN culture medium), serology, PCR and RAPD-PCR, hemogram, and biochemical assays. The dogs presenting VL clinical profile had their lymph nodes and/or bone marrow punctured and their content inoculated in NNN culture medium. The protozoan isolated was identified by PCR and PCR-RAPD. Strains of Leishmania were isolated in 19 out of the 24 studied animals. Fourteen isolates were identified as L. (Leishmania) infantum, and five were L. (Viannia) braziliensis. In the epidemiological analysis, it was possible to determine that all dogs with L. (L.) infantum being allochthonous cases. Leishmaniasis is a zoonose that has the domestic dog as reservoir, the migration of such animals can disseminate the parasite to other regions, provided the agent finds an adequate ecotope and a specific vector (Lutzomyia longipalpis).

Assuntos

RESUMO

O presente trabalho teve como objetivo pesquisar a ocorrência de leishmaniose visceral em cães com sinais clínicos compatíveis, procedentes de clínicas veterinárias das diferentes regiões do Estado do Paraná e em caso positivo verificar a autoctonia dos mesmos. Dos animais com suspeita clínica e epidemiológica de LV, foi coletada amostra de sangue para realização de hemograma, provas bioquímicas, sorologia, cultura do parasito, PCR e RAPD. Os cães portadores de quadro clínico, como febre, hiperqueratose, onicogrifose e emagrecimento exagerado, tiveram gânglio poplíteo e/ou medula óssea puncionados e o conteúdo inoculado em meio de cultivo NNN. A extração de DNA do parasito em sangue e cultura foi realizada pelo método fenol/clorofórmio. A amplificação de DNA do protozoário foi feita por PCR e RAPD. Dos 24 animais analisados, o parasito foi isolado em 19 cães. As técnicas moleculares permitiram identificar 14 isolados como L. (Leihmania) infantum e cinco como L. (Viannia) braziliensis. A análise epidemiológica dos casos permitiu determinar o local de transmissão e definir que todos os cães com LV diagnosticados eram casos alóctones, ou seja eram importados de regiões endêmicas. Sugere-se que a migração dos cães de regiões endêmicas para regiões indenes poderá permitir a instalação de novos focos, desde que o agente encontre ecótopo adequado e vetor específico (Lutzomyia longipalpis).

The south region is the only area in Brazil that does not present autochthonous cases of visceral leishmaniasis (VL), however, in the state of Parana, dogs and humans have been found showing a VL compatible clinical profile. In view of this problem, the present work aimed at isolating and identifying the parasite and determining the cases autochthony. All animals clinically suspect of VL were clinically evaluated, and had samples of their blood collected for hemoculture (NNN culture medium), serology, PCR and RAPD-PCR, hemogram, and biochemical assays. The dogs presenting VL clinical profile had their lymph nodes and/or bone marrow punctured and their content inoculated in NNN culture medium. The protozoan isolated was identified by PCR and PCR-RAPD. Strains of Leishmania were isolated in 19 out of the 24 studied animals. Fourteen isolates were identified as L. (Leishmania) infantum, and five were L. (Viannia) braziliensis. In the epidemiological analysis, it was possible to determine that all dogs with L. (L.) infantum being allochthonous cases. Leishmaniasis is a zoonose that has the domestic dog as reservoir, the migration ofsuch animals can disseminate the parasite to other regions, provided the agent finds an adequate ecotope and a specific vector (Lutzomyia longipalpis).

Assuntos

RESUMO

Com o objetivo de avaliar a ocorrência da infecção por Toxoplasma gondii, em ovinos das regiões periurbana e urbana do município de Curitiba, Paraná, Região Sul do Brasil, foram coletadas amostras de sangue de 167 ovinos, de três rebanhos. O teste de ELISA foi utilizado para detectar anticorpos anti-Toxoplasma gondii. Dos 167 soros ovinos, 43 (25,75%) apresentaram anticorpos anti-T. gondii. O parasito está difundido entre os ovinos das duas regiões estudadas. (AU)

This study evaluated occurrence of the Toxoplasma gondii infection in ovine from urban and periurban areas of Curitiba, State of Paraná, Southern of Brazil. Blood samples from 167 sheep from three farms were collected. ELISA assay was used to detect IgG anti-Toxoplasma gondii antibodies. Of 167 ovine samples, 43 (25.75%) were positive. The parasite is widespread among sheep from urban and periurban areas of Curitiba. (AU)

Assuntos

RESUMO

The present study examines the role that dogs play in the maintenance of the Leishmania cycle in the State of Paraná, Southern Brazil. Dogs were examined in three regions where cutaneous leishmaniasis is endemic or epidemic (R1-Vale da Ribeira; R2-Central region of Paraná State and R3-Northern region). To determine serum prevalence rates ELISA was used. In regions endemic for Trypanosoma cruzi (R1 and R3), serum from dogs seroreactive towards Leishmania antigen was subjected to T. cruzi adsorption in order to eliminate cross-reaction with common antigen epitopes. Concomitantly, dogs with cutaneous lesions were biopsied to isolate and identify parasites using RAPD. Leishmania were classified by the phenetic method using the Jaccard coefficient of similarity, and grouped by Unweighted Pair-Group Method using an Arithmetic Average (UPGMA). A total of 410 dogs were studied. In R1 (Vale da Ribeira) 159 dogs were evaluated of which 10 had anti-Leishmania antibody. In R2 (Central Paraná), 39 animals were examined of which 8 were seropositive. In R3 (the North) 212 dogs were evaluated of which 39 animals were seropositive. Thirteen dogs had cutaneous lesions and the parasites were isolated from a dog with mucocutaneous lesion in R1, two animals with simple skin lesions in R2 and 10 dogs with multiple lesions in R3. The identification of the parasite by molecular methods showed it to be L. (Viannia) braziliensis. Based on this information, the role of domestic dogs in Leishmania infection of cutaneous leishmaniasis in Paraná is discussed.

Assuntos

RESUMO

Studies were carried out on the recovery rate and cysticerci location in bovines experimentally infected with Taenia saginata eggs. Three calves of 6.5 months and one with 19 months of age were infected orally with 2 x 10(4) eggs of Taenia saginata. A fifth calf served as control. After 90 days of infection, the animals were slaughtered and organs and skeletal muscles were inspected using a slicing technique every 5 mm. From the four infected calves, 702 cysticerci were recovered, of which 570 (81.2 percent) were alive and 132 (18.8 percent) were degenerated. The recovery rate ranged from 0.01 to 1.43 percent with an average of 0.88 percent. The cysticerci presented the following anatomical distribution: hioideos muscles 02 (0.28 percent), kidneys 03 (0.43 percent), tongue 07 (1.00 percent), liver 12 (1.71 percent), lungs 15 (2.14 percent), diaphragm 18 (2.56 percent), mastication muscles 25 (3.56 percent), heart 49 (6.98 percent), anterior muscle 323 (46.00 percent) and posterior muscle 248 (35.33 percent)

Assuntos