Ensaios preliminares do Guaiacum officinale L. como moluscicida

Suspensões aquosas do pericarpo do fruto, da casca da raiz, das folhas e das sementes de Guaiacum officinale foram testadas como moluscicida, cercaricida e piscicida em diferentes concentrações. Em laboratório, a suspensão do pericarpo do fruto apresentou 100% de mortalidade a 100 ppm para desovas de B. glabrata, a 20 ppm para caramujos adultos de Biomphalaria glabrata, B. straminea e B. tenagophila, a 5 ppm para Lebistes reticulatus (peixes) e a 1 ppm para cercárias de Schistosoma mansoni. O extrato etanólico do pericarpo do fruto não foi ativo para caramujo adulto de B. glabrata. As doses letais para 90% dos caramujos adultos (DL90), após 24 horas de exposição, usando a suspensão do pericarpo do fruto foram de: 15 ppm para B. glabrata; 14 ppm para B. straminea e 18 ppm para B. tenagophila. As DL90 das suspensões das casca da raiz, sementes e folhas contra B. glabrata foram de 57, 33 e 15 ppm, respectivamente. No campo, coma suspensão do pericarpo do fruto a mortalidade de caramujos adultos de B. glabrata foi de 68% a 20 ppm e 100% a 40 ppm

Sensitivity of an immunoenzymatic test for detection of ant-L. brasiliensis antibodies compared to other tests used for the diagnosis of American cutaneous leishmaniasis

The diagnosis of American cutaneous leishmaniasis (ACL) is frequently based on clinical and epidemiological data associated with the results of laboratory tests. Some laboratory methods are currently being applied for the diagnosis of ACL, among them the indirect immunofluorescence reaction (IIFR), the Montenegro skin test (MST), histopathological examination, and the polymerase chain reaction (PCR). The performance of these methods varies in a considerable proportion of patients. After the standardization of an immunoenzymatic test (ELISA) for the detection of IgG in the serum of patients with ACL using a crude Leishmania braziliensis antigen, the results obtained were compared to those of other tests routinely used for the diagnosis. The tests revealed the following sensitivity, when analyzed separately: 85% for ELISA IgG, 81% for PCR, 64.4% for MST, 58.1% for IIFR, and 34% for the presence of parasites in the biopsy. ELISA was positive in 75% of patients with ACL presenting a negative MST, in 84.8% of ACL patients with negative skin or mucous biopsies for the presence of the parasite, and in 100% of cases with a negative PCR. Thus, ELISA presented a higher sensitivity than the other tests and was useful as a complementary method for the diagnosis of ACL.

Diterpenos de Copaifera reticulata Ducke com atividade larvicida contra Aedes aegypti (L.) (Diptera, Culicidae)

The objective of this study was to evaluate the larvicidal activity of diterpenoids obtained from the oil-resin of Copaifera reticulata against Aedes aegypti larvae, the principal vector of dengue and urban yellow fever. Four diterpenes were obtained from oil-resin extraction with organic solvents and subsequent chromatographic and spectroscopic procedures allowed to isolation and identification of these compounds as 3-b-acetoxylabdan-8(17)-13-dien-15-oic acid (1), alepterolic acid (2), 3-b-hidroxylabdan-8(17)-en-15-oic acid (3), and ent-agatic acid (4). Each compound was previously dissolved in dimethylsulphoxide, and distilled water was added to obtain the desired concentrations. Twenty larvae of third instars were placed into plastic beckers, containing the solution test (25 mL), in a five repetitions scheme, and their mortality, indicated by torpor and darkening of the cephalic capsule, was recorded after 48h. Probit analyses were used to determine lethal concentrations (LC50 and LC90) and their respective 95% confidence intervals. This study showed that only diterpenoids 1 and 2 exhibited larvicidal properties with LC50 of 0.8 ppm and 87.3 ppm, respectively, revealing the former as the most toxic compound against third instars of Ae. aegypti. Therefore, this compound seems to be an interesting source for new metabolite to be exploited.

L. (L.) chagasi in aids and visceral leishmaniasis (kala-azar) co-infection

Concomitant skin lesions in visceral leishmaniasis (VL) or kala-azar are rare, being more common the description of post-kala-azar dermal leishmaniasis occurring post treatment of kala-azar. Skin lesions caused by Leishmania donovani are frequently seen in the aids-VL co-infection. In Brazil cutaneous or mucosal forms of tegumentary leishmaniasis concomitant with aids are more commonly registered. Here we present a case of aids-VL co-infection, with unusual cutaneous and digestive compromising attributed to L. (L.) chagasi, with special attention to ecthymatous aspect of the lesion, allied to the absence of parasite on the histological skin biopsy.

Immunoperoxidase technique using an anti-Leishmania (L.) chagasi hyperimmune serum in the diagnosis of culture-confirmed American tegumentary leishmaniasis

The present study reports the production of the rabbit anti-Leishmania (L.) chagasi hyperimmune serum, the standardization of the immunohistochemistry (IHC) technique and the evaluation of its employment in cutaneous leishmaniasis (CL) lesions diagnosed by Leishmania sp. culture isolation. Thirty fragments of active CL lesions were examined as well as 10 fragments of cutaneous mycosis lesions as control group. IHC proved more sensitive in detecting amastigotes than conventional hematoxylin-eosin (HE) stained slides: the former was positive in 24 (80%) biopsies whereas the latter, in 16 (53%) (p = 0.028). The reaction stained different fungus species causing cutaneous mycosis. Besides, positive reaction was noticed in mononuclear and endothelial cells. Nevertheless, this finding was present in the control group biopsies. It is concluded that IHC showed good sensitivity in detecting amastigotes.

Canine visceral leishmaniasis due to Leishmania (L.) infantum chagasi in Amazonian Brazil: comparison of the parasite density from the skin, lymph node and visceral tissues between symptomatic and asymptomatic, seropositive dogs

Canine visceral leishmaniasis (CVL) is recognizable by characteristic signs of disease and is highly lethal. The infection, however, may be quite inapparent in some seropositive dogs, and this has raised the polemic question as to whether or not such animals can be a source of infection for Lutzomyia longipalpis, the vector of American visceral leishmaniasis (AVL). In this study we have examined 51 dogs with acute CVL from an AVL area in Pará State, northern Brazil, and compared the parasite density, amastigotes of Leishmania (L.) infantum chagasi, in the skin, lymph node and viscera of symptomatic with that of nine asymptomatic but seropositive dogs (IFAT-IgG). Post-mortem biopsy fragments of these tissues were processed by immunohistochemistry, using a polyclonal antibody against Leishmania sp. The X² and Mann Whitney tests were used to evaluate the means of infected macrophage density (p < 0.05). There was no difference (p > 0.05) in the skin (10.7/mm² x 15.5/mm²) and lymph node (6.3/mm² x 8.3/mm²), between asymptomatic and symptomatic dogs, respectively. It was higher (p < 0.05), however, in the viscera of symptomatic (5.3/mm²) than it was in asymptomatic (1.4/mm²) dogs. These results strongly suggest that asymptomatic or symptomatic L. (L.) i. chagasi-infected dogs can serve as a source of infection, principally considering the highest (p < 0.05) parasite density from skin (10.7/mm² x 15.5/mm²), the place where the vetor L. longipalpis takes its blood meal, compared with those from lymph node (6.3/mm² x 8.3/mm²) and viscera (1.4/mm²x 5.3/mm²).

Susceptibility of Cebus apella monkey (Primates: Cebidae) to experimental Leishmania (L.) infantum chagasi-infection

In Amazonian Brazil, the Cebus apella monkey (Primates: Cebidae) has been associated with the enzootic cycle of Leishmania (V.) shawi, a dermotropic parasite causing American cutaneous leishmaniasis (ACL). It has also been successfully used as animal model for studying cutaneous leishmaniasis. In this work, there has been investigated its susceptibility to experimental Leishmania (L.) infantum chagasi-infection, the etiologic agent of American visceral leishmaniasis (AVL). There were used ten C. apella specimens, eight adult and two young, four males and six females, all born and raised in captivity. Two experimental infection protocols were performed: i) six monkeys were inoculated, intra-dermal via (ID), into the base of the tail with 2 x 10(6) promastigotes forms from the stationary phase culture medium; ii) other four monkeys were inoculated with 3 x 10(7) amastigotes forms from the visceral infection of infected hamsters by two different via: a) two by intravenous via (IV) and, b) other two by intra-peritoneal via (IP). The parameters of infection evaluation included: a) clinical: physical exam of abdomen, weigh and body temperature; b) parasitological: needle aspiration of the bone-marrow for searching of amastigotes (Giemsa-stained smears) and promastigotes forms (culture medium); c) immunological: Indirect fluorescence antibody test (IFAT) and, Delayed-type hypersensitivity (DTH). In the six monkeys ID inoculated (promastigotes forms) all parameters of infection evaluation were negative during the 12 months period of follow-up. Among the four monkeys inoculated with amastigotes forms, two IV inoculated showed the parasite in the bone-marrow from the first toward to the sixth month p.i. and following that they cleared the infection, whereas the other two IP inoculated were totally negative. These four monkeys showed specific IgG-antibody response since the third month p.i. (IP: 1/80 and IV: 1/320 IgG) toward to the 12th month (IP: 1/160 and IV: 1/5120). The DTH-conversion occurred in only one IV inoculated monkey with a strong (30 mm) skin reaction. Considering these results, we do not encourage the use of C. apella monkey as animal model for studying the AVL.

Susceptibility of peritoneal macrophage from different species of neotropical primates to Ex vivo Leishmania (L.) infantum chagasi-infection

This study examined the susceptibility of peritoneal macrophage (PM) from the Neotropical primates: Callithrix jacchus, Callithrix penicillata, Saimiri sciureus, Aotus azarae infulatus and Callimico goeldii to ex vivo Leishmania (L.) infantum chagasi-infection, the etiological agent of American visceral leishmaniasis (AVL), as a screening assay for evaluating the potential of these non-human primates as experimental models for studying AVL. The PM-susceptibility to infection was accessed by the PM-infection index (PMI) at 24, 72 h and by the mean of these rates (FPMI), as well as by the TNF-α, IL-12 (Capture ELISA) and Nitric oxide (NO) responses (Griess method). At 24h, the PMI of A. azarae infulatus (128) was higher than those of C. penicillata (83), C. goeldii (78), S. sciureus (77) and C. jacchus (55). At 72h, there was a significant PMI decrease in four monkeys: A. azarae infulatus (128/37), C. penicillata (83/38), S. sciureus (77/38) and C. jacchus (55/12), with exception of C. goeldii (78/54). The FPMI of A. azarae infulatus (82.5) and C. goeldii (66) were higher than C. jacchus (33.5), but not higher than those of C. penicillata (60.5) and S. sciureus (57.5). The TNF-a response was more regular in those four primates which decreased their PMI at 24/72 h: C. jacchus (145/122 pg/mL), C. penicillata (154/130 pg/mL), S. sciureus (164/104 pg/mL) and A. azarae infulatus (154/104 pg/mL), with exception of C. goeldii (38/83 pg/mL). The IL-12 response was mainly prominent in A. infulatus and C. goeldii which presented the highest FPMI and, the NO response was higher in C. goeldii, mainly at 72 h. These findings strongly suggest that these New World primates have developed a resistant innate immune response mechanism capable of controlling the macrophage intracellular growth of L. (L.) i. chagasi-infection, which do not encourage their use as animal model for studying AVL.

Molluscicidal and Mosquitocidal Activities of the Essential oils of Thymus capitatus Hoff. et Link. and Marrubium vulgare L.

Steam distillation of essential oils of aerial parts of Thymus capitatus and Marrubium vulgare L. collected at North cost of Egypt yielded 0.5% and 0.2%, respectively. Results of Gas chromatography-mass spectrometry analyses of the two samples identified 96.27% and 90.19% of the total oil composition for T. capitatus and M. vulgare, respectively. The two oil samples appeared dominated by the oxygenated constituents (88.22% for T. capitatus and 57.50% for M. vulgare), composed of phenols, mainly carvacrol (32.98%) and thymol (32.82%) in essential oil of T. capitatus, and thymol (34.55%) in essential oil of M. vulgare. It was evaluated the molluscicidal activity of T. capitatus and M. vulgare essential oils on adult and eggs of Biomphalaria alexandrina as well as their mosquitocidal activity on Culex pipiens. The LC50 and LC90 of T. capitatus essential oil against adult snails was 200 and 400 ppm/3hrs, respectively, while for M. vulgare it was 50 and 100 ppm/3hrs, respectively. Moreover, M. vulgare showed LC100 ovicidal activity at 200 ppm/24 hrs while T. capitatus oil showed no ovicidal activity. It was verified mosquitocidal activity, with LC50 and LC90 of 100 and 200 ppm/12hrs respectively for larvae, and 200 and 400 ppm/12hrs respectively for pupae of C. pipiens.

ANTIFUNGAL ACTIVITY OF Cymbopogon nardus (L.) Rendle (CITRONELLA) AGAINST Microsporum canis FROM ANIMALS AND HOME ENVIRONMENT

Dermatophytosis is a common zoonosis in urban centers. Dogs and cats have played an important role as its disseminators. Environmental decontamination is essential for the prevention of its propagation to humans and animals. However, sanitizers or disinfectants with antifungal activity, currently available, have high toxicity. The present study evaluated the in vitro effects of an extract of citronella (Cymbopogon nardus) on 31 Microsporum canis isolates from animals and home environments. Susceptibility tests were performed based on document M38-A2 (2008) of the Clinical and Laboratory Standards Institute with modifications for natural products. Although susceptibility variation was observed between the fungus tested, the concentrations that inhibited the growth of 50 and 90% of the microorganisms were low (19.5 and 78 µg/mL, respectively). Thus, this citronella extract showed potent fungistatic and fungicide activities against M. canis isolated from animals and home environments. Therefore, it could be an alternative for dermatophytosis prophylaxis in the home environment.

Estudos sôbre reservatórios e vetores silvestres do Trypanosoma cruzi: XVII. Contribuição para o estudo dos focos naturais da tripanossomose americana, com especial referência à região nordeste do Estado de São Paulo, Brasil

Depois de acentuar que a tripanossomose americana é uma zoonose do tipo anfixenose, bem enquadrável no conceito de PAVLOVSKY de infecção com focos naturais, o Autor analisa o problema da multiplicidade e diversidade dêstes focos que são devidas ao grande número de hospedeiros e vetores naturais com hábitos variados. Descreve, em seguida, alguns focos naturais mais freqüentes e importantes, observados na região nordeste do Estado de São Paulo e áreas limítrofes do Estado de Minas Gerais, focos êstes constituídos por buracos e cavidades no solo, ocos e anfractuosidades em troncos de árvores, tufos de vegetação herbácea, touceiras de piteira e copa de palmeiras, onde triatomíneos e mamíferos convivem.

Estudos sôbre reservatórios e vetores silvestres do Trypanosoma cruzi. XXII. Modificações dos focos naturais da tripanossomose americana e suas conseqüências

Os focos naturais da tripanossomose americana, como tôda biogeocenose, podem. permanecer relativamente estáveis, sendo esta estabilidade necessária para a própria existência dêsses focos. Mas o balanço ecológico pode ser alterado, sendo muito importante aquelas alterações devidas à interferência direta ou indireta do homem ao exercer atividades ligadas ao desbravamento e à colonização. Entre as conseqüências podemos citar: 1) redução ou desaparição dos focos naturais; 2) concentração de hospedeiros e vectores em áreas favoráveis limitadas; 3) deslocamento de hospedeiros e vectores para outras áreas; 4) invasão das habitações humanas e anexos, com a instalação do ciclo doméstico da infecção. Estas possibilidades são analisadas e ilustradas com exemplos.

Tratamento da leishmaniose tegumentar americana pelo niridazol

Foi empregue o niridazol um derivado do nitrotiazol, em doze pacientes com leishmaniose tegumentar americana. Em todos êles, o diagnóstico clínico da doença foi confirmado pela biópsia das lesões e em oito dêles também pela positividade da intradetermorreação de Montenegro. Oito pacientes tinham lesões mucosas metastáticas em atividade e lesões cutâneas iniciais cicatrizadas há tempo mais ou menos longo. Dois apresentavam concomitância de lesões cutâneas e mucosas em atividade, e dois tinham lesões cutâneas exclusivas. A duração da doença variou de 2 meses a 32 anos. A posologia diária do niridazol foi uniformememente de 25 mg/kg de pêso corpóreo. O medicamento foi administrado por via oral, em duas tomadas diárias, sempre com o doente internado em hospital. Quando a tolerância o permitia, o paciente recebia cinco séries de tratamento de 10 dias de duração cada, intercaladas por períodos áe suspensão da droga de 10 dias entre uma série e outra. Isso foi possível em 10 dos 12 pacientes. O tratamento foi bem tolerado em 5 doentes e de tolerância regular em 4. Mal tolerado em um paciente pela ocorrência de alucinações e excitação mental e interrompido pela péssima tolerância em dois doentes devido ao aparecimento de convulsões generalizadas com perda da consciência. O tratamento acompanhou-se de grande incidência, de efeitos colaterais. A ocorrência de para-efeitos mais intensos não foi devida às más condições hepáticas. Provas de função hepática. hemogramas, estudos bioquímicos do sangue e exames de urina, realizados antes, durante e após o tratamento, não revelaram alterações significativas. Biópsias hepáticas por punção com agulha em dez doentes, prévias e posteriores ao tratamento, não detectaram lesões hepáticas que pudessem ser atribuídas à medicação. O seguimento dos doentes prolongou-se pelo prazo de dois a 36 meses apos o tratamento. Alterações eletrocardiográficas foram detectadas em sete de oito doentes que não tinham cardiopatias concomitantes (87,5% dos casos), mas não se acompanharam de clínica relacionada com o aparelho cardiovascular. Alterações eletroencefalográficas foram observadas em 3 de 9 doentes que foram submetidos a exames seriados. O autor ficou decepcionado com os maus resultados terapêuticos. A droga curou as lesões cutâneas de dois dentre três doentes nos quais puderam ser completadas as cinco séries de tratamento. A temporária e aparente melhora das lesões mucosas seguiu-se sempre da sua recidiva, após o término do tratamento em todos os pacientes com lesões mucosas. O autor conclui que o niridazol não é medicamento eficaz na terapêutica da leishmaniose tegumentar americana com lesões mucosas metastáticas tardias localizadas no nariz, boca e/ou faringe. O autor aconselha vigilância dos doentes durante o tratamento devido à possibilidade do aparecimento de manifestações neuropsíquicas.

Pequeno surto de leishmaniose tegumentar americana em Macuco (Cordeiro - RJ)

Por ocasião de um pequeno surto de leishmaniose tegumentar americana em localidade do município de Cordeiro (RJ) foram documentados 14 casos. Entre os fatos que chamaram a atenção dos autores estão a alta ocorrência em crianças (57%), a freqüência do componente linfangítico (40%), a tendência à cicatrização espontânea e a distribuição peri-domiciliar dos casos. Um inquérito pela intradermoreação de Montenegro na área de maior concentração de casos, realizado numa fazendo e numa escola, mostrou 17% de reações positivas, 17% de duvidosas e 66% de reações negativas, num total de 117 indivíduos testados. Não foram encontrados casos antigos cicatrizados, o que sugere um foco recente. Discute-se o significado das reações duvidosas.