Estudo experimental de um "Schizotrypanum" do morcego "Hemiderma perspicillatum" da Venezuela

1) - Em Guarativana, Estado de Yaracuy, Venezuela, foi determinada a infecção natural de morcegos da espécie Hemiderma perspicillatum (L) por um hemoflagelado do gênero Schizotrypanum CHAGAS. 2) - As formas sanguícolas do parasito são morfologicamente muito semelhantes às do Trypanosoma phyllostomae CARTAYA, 1910, descrito em morcegos da mesma espécie, em Cuba. Sem por ora identificarmos a este parasito o que encontramos na Venezuela, continuaremos a referir-nos a este último como amostra phyllostomae (cf. DIAS, VTDJ). 3) - Os tripanosomas da amostra phyllostomae teem em média 20µ de comprimento total, possuem volumoso blefaroplasto subterminal, núcleo elíptico quese mediano, porem mais próvimo à extremidade anterior do corpo. As seguintes médias foram obtidas da medida de 100 tripanosomas: Extremidade posterior ao meio do núcleo 7,1µ; Meio do núcleo à extremidade anteior 5,0µ; Flagelo livre 7,8µ; Comprimento total 20,0µ; Posição do núcleo (índice PN/NA) 1,4. A amostra phyllostomae mostrou-se biometricamente diferençavel de todas as demais de Schizotupanum estudadas (DIAS & FREITAS). 4) - Formas de multiplicação com a morfologia de leishmania foram encontradas em coração e estômago de morcegos infectados. Tripanosomas em via de divisão nunca foram observados no sangue. 5) - Cobaia, cão e camondongo branco são sensiveis à amostra, sofrendo algumas vezes infecções intensas e mortais. Leishmanias intracelulares e lesões foram encontradas em diversos orgãos, principalmente no coração. 6) - O morcego Phyllostomae hastatus não é sensivel à amostra, apresentando apenas um parasitismo sanguíneo escasso e transitório quando inoculado e reinoculado repetidas vezes. O Molossus obscurus tambem é aparentemente insensivel, enquanto que os Molossus rufus parece receptivel ao parasito. 7) - Após numerosas passagens por animais de laboratório, a amostra Phyllostomae foi infesctante para dois exemplares de Hemiderma perspicillatum do Brasil. Os morcegos H. perpicillatum e H. perspicillatum aztecum são aparentemente resistentes á inoculação do Schizotrypanum cruzi (tipo humano). 8) - A amostra phyllostomae evoluiu facilmente em todos os artrópodes sugadores experimentados: Rhodnius prolixus, Eutriatoma nigromaculata, E. maculata, E. sordida, Panstrongylus megistus, P. geniculatus, Triatoma infestans,Psammolestes arthuri, Cimex hemipterus e Ornithodoros moubata. A evolução processa-se analogamente à do S. chuzi, terminando com a formação de tripanosomas metacíclicos no intestino posterior. 9) - Cultivos artificiais são facilmente conseguidos em meios apropriados. 10) - A amostra phyllostomae distingue-se biológica e morfologicamente de outros hemoflagelados de morcegos (amostras hastatus e vespertilionis). Biologicamente aproxima-se muito do Schizotrydanum cruzi humano, do qual é, entretanto, diferençavel por processos biométricos (DIAS, 1940, DIAS & FREITAS).

Clearing factor pancreático

Dosou-se o "clearing factor" (lipase lipoproteica), em pâncreas de ratos normais após serem submetidos a algumas condições experimentais. As seguintes conclusões podem ser extraídas: a) O pãncreas é o órgão mais rico nesta enzima (8 vezes mais que o tecido adiposo, até então considerado o mais rico em CF); b) O jejum total de 48 horas diminui significamente a atividade de CF; c) O complamin, o derivado do ácido nicotínico, agindo sobre a arteriosclerose experimental, não afeta a atividade da enzima; d) O ácido épsilon-aminocapróico (AEAC) potente antagonista da heparina, inibe a ativdade do Cf pancreático; e) Ratos com arteriosclerose experimental apresentam níveis diminuídos desta enzima no pâncreas.

Jatobal virus antigenic characterization by ELISA and neutralization test using EIA as indicator, on tissue culture

A virus antigenic characterization methodology using an indirect method of antibody detection ELISA with virus-infected cultured cells as antigen and a micro virus neutralisation test using EIA (NT-EIA) as an aid to reading were used for antigenic characterization of Jatobal (BeAn 423380). Jatobal virus was characterized as a Bunyaviridae, Bunyavirus genus, Simbu serogroup virus. ELISA using infected cultured cells as antigen is a sensitive and reliable method for identification of viruses and has many advantages over conventional antibody capture ELISA's and other tests: it eliminates solid phase coating with virus and laborious antigen preparation; it permits screening of large numbers of virus antisera faster and more easily than by CF, HAI, or plaque reduction NT. ELISA and NT using EIA as an aid to reading can be applicable to viruses which do not produce cytopathogenic effect. Both techniques are applicable to identification of viruses which grow in mosquito cells.

Isolation of Pseudomonas cepacia in cystic fibrosis patient

Pulmonary infection on cystic fibrosis (CF) patients are associated with a limited qualitative number of microorganisms. During the colonization process, Staphylococcus aureus usually preceedes Pseudomonas aeruginosa. This latter is at first non-mucoid, being replaced or associated to a mucoid morphotype which is rare in other diseases. In 1980, Pseudomonas cepacia appeared as an important agent in CF pulmonary infections with a mean frequency of about 6.1% isolations in different parts of the world. The primus colonization mainly occurs in the presence of pre-existent tissue lesions and the clinical progress of the disease is variable. In some patients it can be fulminant; in others it can cause a gradual and slow decrease in their pulmonary functions. The concern with this germ isolation is justified by its antibiotic multiple resistence and the possibility of direct transmission from a colonized patient to a non-colonized one. We reported the first case of P. cepacia infection in a CF patient in our area. The microbiological attendance to this patient had been made from 1986 to 1991 and the first positive culture appeared in 1988. The sensitivity profile showed that the primus colonization strain was sensitive to 9 of 17 tested antibiotics, however in the last culture the strain was resistent to all antibiotics. These data corroborate the need for monitoring the bacterial flora on CF patients respiratory system.

Use of PCR for the determination of the frequency of the deltaF508 mutation in Brasilian cistic fibrosis patients

The [Delta]F508 mutation in the cystic fibrosis (CF) gene was studied in a population of 18 Brazilian CF patients and their 17 families by use of PCR and differential hybridization with oligonucleotides. In a total of 34 chromosomes considered, 12 (35%) carried the F508 deletion, a frequency much lower than that reported in most other populations. As a consequence, CF in Brazil would be predominantly caused by mutations different from the F508 deletion

Standardization of serological tests for detecting anti-Trypanosoma cruzi antibodies in dogs

This paper reports on the standardization of four serological reactions currently used in human serodiagnosis for the detection of anti-Trypanosoma cruzi antibodies in naturally and experimentally infected dogs. Indirect immunofluorescence test (IFAT) and hemagglutination test (IHAT) were standardized, and complement fixation test (CFT) and direct agglutination test (DAT) were used for diagnostic confirmation. Four hundred and eighty one mongrel dogs that were studied by xenodiagnosis were used: (1) parasitemic dogs of two localities of endemic area (EA) of Santiago del Estero province in Argentina (n = 134); (2) non-parasitemic dogs of the same area (n = 285); (3) dogs experimentally infected with T. cruzi in the patent period (n = 6); (4) non-infected dogs (n = 56) which were born in the city of Buenos Aires (BA), one non-EA for Chagas' disease. For IFAT, parasitemic dogs EA showed 95% of reactive sera. Non parasitemic dogs EA showed 77% of non reactive sera. None sera from BA were reactive for dilutions higher than four. For IHAT, 84% of sera of parasitemic dogs EA showed serological reactivity and among non parasitemic dogs BA, 61% were non reactive, while the remainder showed at most titres of 1/16. The cut-off titres for IFAT and IHAT were 1/16 and 1/32 respectively, and for CFT and DAT 1/1 and 1/128 respectively. Sensitivity for IFAT, IHAT, CF and DAT were 95%, 84%, 97% and 95% respectively.

Neotropical Monogenoidea. 23. Two new species of Gyrodactylus (Gyrodactylidae) from a Cichlid and an Erythrinid fish of Southeastern Brazil

Two new species of Gyrodactylus Nordmann, 1832 (Platyhelminthes, Monogenoidea) are described from fishes collected from southeastern Brazil. Gyrodactylus geophagensis n. sp. was collected from the body surface of the "cará", Geophagus brasiliensis (Quoy and Gaimard, 1824) (Cichlidae), from the Rio da Guarda, state of Rio de Janeiro; its major diagnostic features are the morphology of the anchor with a short, truncate superficial root and the shape of the hooks - with a long, delicate shaft. Gyrodactylus trairae n. sp. parasitizes the body surface of the "traíra", Hoplias aff. malabaricus (Bloch, 1794) (Erythrinidae), from the rio Guandu, state of Rio de Janeiro and can be easily differentiated from other species of the genus by having a thin, dorsal bridge, connecting the superficial bar with the spathulated shield. These are the first species of Gyrodactylus formally reported from Brazil. Presently, 26 species of Gyrodactylidae are known from freshwater fishes in the neotropical region; a list of these species is provided.

Tumor necrosis factor-α in human American tegumentary leishmaniasis

Tumor necrosis factor-alpha (TNF-α) is a cytokine produced by activated macrophages and other cells. In order to verify whether the serum levels of TNF-α in American tegumentary leishmaniasis patients are associated with the process of cure or aggravation of the disease, 41 patients were studied: 26 cases of cutaneous leishmaniasis (CL) and 15 of mucocutaneous leishmaniasis (MCL). During active disease the serum levels of TNF-α of MCL patients were significantly higher than those of CL patients and control subjects (healthy individuals and cutaneous lesions from other etiologies). The MCL patients had serum titers of TNF-α significantly lower at the end of antimonial therapy than before therapy. After a six-month follow-up, the MCL patients had serum levels of TNF-α similar to those observed at the end of the therapy as well as to those of CL patients and control subjects. No significant variation in the serum levels of TNF-α was observed in CL patients throughout the study period (before, at the end of therapy and after a six-month follow-up). The possible relationship between the high TNF-α serum levels and severity of the disease is discussed.

Prognosis of schistosomiasis mansoni patients infected with hepatitis B virus

A clinical study on the evolution of patients with schistosomiasis mansoni has been conducted since 1983 at the outpatient clinic of the Infectious and Parasitic Disease Service in the Clementino Fraga Filho University Hospital in Rio de Janeiro, Brazil, comparing prevalence of positive tests for HBsAg, anti-HBsAg, and anti-HBc among patients infected with Schistosoma mansoni coming from various regions of Brazil and with different clinical forms of the disease. A non-significant predominance of HBsAg, anti-HBsAg, and anti-HBc was detected among patients with the hepatosplenic form of schistosomiasis, who presented a more severe clinical evolution with a higher frequency of hematemesis and/or melena, in addition to the development of macronodular cirrhosis and a worse prognosis as compared to patients with the toxemic form, schistosomiasis-infection and the hepatointestinal form.

Atypical Mucocutaneous Leishmaniasis Caused by Leishmania braziliensis in an Acquired Immunodeficiency Syndrome Patient: T-cell Responses and Remission of Lesions Associated with Antigen Immunotherapy

An atypical case of acquired immunodeficiency syndrome-associated mucocutaneous lesions due to Leishmania braziliensis is described. Many vacuolated macrophages laden with amastigote forms of the parasite were found in the lesions. Leishmanin skin test and serology for leishmaniasis were both negative. The patient was resistant to therapy with conventional drugs (antimonial and amphotericin B). Interestingly, remission of lesions was achieved after an alternative combined therapy of antimonial associated with immunotherapy (whole promastigote antigens). Peripheral blood mononuclear cells were separated and stimulated in vitro with Leishmania antigens to test the lymphoproliferative responses (LPR). Before the combined immunochemotherapy, the LPR to leishmanial antigens was negligible (stimulation index - SI=1.4). After the first course of combined therapy it became positive (SI=4.17). The antigen responding cells were predominantly T-cells (47.5%) most of them with CD8+ phenotype (33%). Very low CD4+ cells (2.2%) percentages were detected. The increased T-cell responsiveness to leishmanial antigens after combined therapy was accompanied by interferon-g (IFN-g) production as observed in the cell culture supernatants. In this patient, healing of the leishmaniasis lesions was associated with the induction of a specific T-cell immune response, characterized by the production of IFN-g and the predominance of the CD8+ phenotype among the Leishmania-reactive T-cells.

Studies on the effectiveness of diarylheptanoids derivatives against Leishmania amazonensis

In a previous work we demonstrated that diarylheptanoids extracted from Centrolobium sclerophyllum are very active against Leishmania amazonensis promastigotes. In order to continue our studies with these class of compounds, we decided to evaluate the activity of several diarylheptanoids derived from curcumin (diferuloyl methane) against the extracellular form (promastigotes) of L. amazonensis. Furthermore, an experiment against the intracellular form of the parasite (amastigotes) was carried out, comparing the most active compound among the curcumin derivatives (the methylcurcumin) with des-O-methylcentrolobine, the most active diarylheptanoid derived from C. sclerophyllum.

TT virus infection in children and adults who visited a general hospital in the south of Brazil for routine procedure

TT virus (TTV) is a newly described nonenveloped human virus, with a circular, negative-stranded DNA genome, that was first identified in the blood of a patient with posttransfusion hepatitis of unknown etiology. PCR primers and conditions used for TTV DNA amplification may greatly influence the level of TTV detection in serum. Three PCR assays, with different regions of the genome as targets, were used to test TTV DNA in 130 sera from children and adults visiting a hospital in the south of Brazil, most of them for routine procedure. Forty-four percent of adult sera and 73% of sera from children aged 0-10 years were TTV positive with at least one PCR assay. However, the three assays were able to detect only 33%, 35%, and 70% of the total positive samples. Our results showed a high prevalence of TTV infection in the south of Brazil, particularly among young children, and confirmed the necessity of performing several PCR assays to assess the true TTV prevalence in a determined population.

Screening of Brazilian basidiomycetes for antimicrobial activity

A total of 103 isolates of basidiomycetes, representing 84 species from different Brazilian ecosystems, were evaluated for their antifungal and antibacterial activity in a panel of pathogenic and non-pathogenic microorganisms. Tissue plugs of the fruiting bodies were cultivated in liquid media and the whole culture extracted with ethyl acetate. Crude extracts from Agaricus cf. nigrecentulus, Agrocybe perfecta, Climacodon pulcherrimus, Gloeoporus thelephoroides, Hexagonia hydnoides, Irpex lacteus, Leucoagaricus cf. cinereus, Marasmius cf. bellus, Marasmius sp., Nothopanus hygrophanus, Oudemansiella canarii, Pycnoporus sanguineus, Phellinus sp., and Tyromyces duracinus presented significant activity against one or more of the target microorganisms. Eight isolates were active only against bacteria while three inhibited exclusively the growth of fungi. Two extracts presented wide antimicrobial spectrum and were active against both fungi and bacteria. Differences in the bioactivity of extracts obtained from isolates from the same species were observed.