Imatinib, a drug used for treatment of human chronic myeloid leukaemia, due to its activity against protein kinases, has been also evaluated in vitro against Schistosoma mansoni showing high schistosomicidal activity. In the present experiments imatinib activity in vitro was confirmed at the doses of 25 µM, 50 µM and 100 µM. The first drug activity observed with the lower dose was interruption of egg-laying and with the higher dosages was the death of the worms. In mice infected with S. mansoni no activity was found even with 1,000 mg/kg/day, 500 mg/kg/day, single oral dose or when administered for three consecutive days. This is another example of the difference of results related to in vitro and in vivo trials using S. mansoni worms.

Mice infected with Schistosoma mansoni were treated with oxamniquine, praziquantel, artesunate at the pre-patent phase, aiming at observing schistogram alterations. Half of the animals were perfused five days post-treatment for counting and classification of immature worms, based on pre-established morphological criteria (schistogram); the remaining animals were evaluated 42 or 100 days after infection and perfusion of the portal-system was performed for collection and counting of adult worms and oogram. It was observed that oxamniquine and artesunate treatment administered at the pre-postural phase causes significant reduction in the number of immature and adult worms. However, there was little reduction with praziquantel when used at the dose of 400 mg/kg for treatments administered 14, 15, 21 or 23 days post-infection. Artesunate was responsible for significant alterations in development of young worms, as well as for a higher number of worms presenting intestinal damages. Immature adult worms were detected in mice treated with artesunate or oxamniquine at the pre-patent phase of infection and recovered by perfusion 100 days after infection. Schistogram proved to be a very useful tool for experimental evaluation of the activity of antischistosomal drugs and a good model to identify the most sensitive stages to drugs.

Current schistosomiasis control strategies are largely based on chemotherapeutic agents and a limited number of drugs are available today. Praziquantel (PZQ) is the only drug currently used in schistosomiasis control programs. Unfortunately, this drug shows poor efficacy in patients during the earliest infection phases. The effects of PZQ appear to operate on the voltage-operated Ca2+channels, which are located on the external Schistosoma mansoni membrane. Because some Ca2+channels have dihydropyridine drug class (a class that includes nifedipine) sensitivity, an in vitro analysis using a calcium channel antagonist (clinically used for cardiovascular hypertension) was performed to determine the antischistosomal effects of nifedipine on schistosomula and adult worm cultures. Nifedipine demonstrated antischistosomal activity against schistosomula and significantly reduced viability at all of the concentrations used alone or in combination with PZQ. In contrast, PZQ did not show significant efficacy when used alone. Adult worms were also affected by nifedipine after a 24 h incubation and exhibited impaired motility, several lesions on the tegument and intense contractility. These data support the idea of Ca2+channels subunits as drug targets and favour alternative therapeutic schemes when drug resistance has been reported. In this paper, strong arguments encouraging drug research are presented, with a focus on exploring schistosomal Ca2+channels.

To elucidate the mechanisms of antischistosoma resistance, drug-resistant Schistosoma mansoni laboratory isolates are essential. We developed a new method for inducing resistance to praziquantel (PZQ) using successive drug treatments of Biomphalaria glabrata snails infected with S. mansoni. Infected B. glabrata were treated three times with 100 mg/kg PZQ for five consecutive days with a one-week interval between them. After the treatment, the cercariae (LE-PZQ) produced from these snails and the LE strains (susceptible) were used to infect mice. Forty-five days after infection, mice were treated with 200, 400 or 800 mg/kg PZQ. Thirty days post-treatment, we observed that the mean number of worms recovered by perfusion was significantly higher in the group of mice infected with the LE-PZQ isolate treated with 200 and 400 mg/kg in comparison to the LE strain with the same treatment. Moreover, there was a significant difference between the ED50 (effective dose required to kill 50% of the worms) of the LE-PZQ isolate (362 mg/kg) and the LE strain (68 mg/kg). In the in vitro assays, the worms of the LE-PZQ isolate were also less susceptible to PZQ. Thus, the use of infected snails as an experimental model for development of resistance to S. mansoni is effective, fast, simple and cheap.

The activity of lovastatin associated with oxamniquine or praziquantel against schistosomiasis mansoni was evaluated in mice infected with Schistosoma mansoni. Forty days after infection, mice were treated with lovastatin, 400 mg/kg for five consecutive days by oral route, and on the last day of this sequence with 50 mg/kg oxamniquine or with 200 mg/kg praziquantel, both by oral route, single dose. Fifteen days later, the animals were perfused in parallel with an untreated control group. Studies were carried out in vitro, using lovastatin in culture medium containing S. mansoni worms proceeding from experimentally infected mice. In the in vivo trials, the association of lovastatin with oxamniquine or praziquantel did not show any additive action, but there were oogram changes when lovastatin was associated with oxamniquine. In vitro lovastatin was able to interrupt the maturation of S. mansoni eggs, which remained at the 1st or 2nd stages, depending on the dose used. The total number of morphologically dead eggs found in culture of worms exposed to 2 µg/ml or 4 µg/ml concentrations of lovastatin was significantly higher than the number of viable eggs. Using the probe Hoescht 33258 it was observed that 70% of the eggs considered morphologically viable in the treated groups (against 16% in the control group) were labeled, indicating that the majority of the viable eggs had membrane permeability increased due to lovastatin action.

Com o objetivo de avaliar a ação da lovastatina na postura do Schistosoma mansoni foram usados camundongos infectados com 100 ±10 cercárias da cepa LE. Trinta dias após a infecção, os animais foram tratados com 100, 200 e 400mg/kg de lovastatina, via oral, durante cinco dias consecutivos e sacrificados 7,15,30 ou 60 dias após o tratamento. Foram analisados: distribuição de vermes no mesentério e fígado, mortalidade de vermes no fígado, alteração do oograma, contagem de ovos no jejuno e fígado, presença de ovos intra-uterino e morfologia dos vermes dos grupos tratados e controle (animais infectados e não tratados). Diferenças estatisticamente significativas foram encontradas, na dose de 400mg/kg, entre grupos tratados e controles quando se considerou a presença de ovos no útero, a alteração do oograma, ovos nos diferentes estágios de desenvolvimento no jejuno e fígado e no comprimento do corpo dos vermes machos e fêmeas. O estudo morfológico dos vermes mostrou alterações degenerativas sendo as principais no aparelho reprodutor, com redução e alteração dos folículos vitelínicos e do ovário das fêmeas e modificações nos testículos dos machos. Os resultados apresentados levam à conclusão que a droga em estudo reduz, consideravelmente, a oviposição das fêmeas do S. mansoni, aumenta o tamanho dos vermes, provoca alterações no sistema reprodutivo de machos e fêmeas e pode ocasionar morte de parte significativa da população de vermes na dose de 400mg/kg.

Aimed to evaluate the lovastatin action on Schistosoma mansoni oviposition, infected mice with 100 ± 10 cercariae of the LE strain were used. Thirty days after infection the animals were treated with 100, 200 and 400mg/kg of lovastatin, per os, during five consecutive days and then sacrificed 7, 15, 30 or 60 days after treatment. We analyzed: distribution of worms in mesenteries and liver; mortality of worms in the liver; alteration of the oogram; eggs counting in the jejunum and liver; presence of intrauterine eggs and morphology of the worms from the treated and control groups (infected and not treated animals). Significant statistical differences were found between treated and control groups when the presence of intrauterine eggs was considered and also, alteration on the oogram, eggs at different stages of development in jejunum, liver and in the body length of males and females. The morphological study of the worms showed that the degenerative modifications occurred, mainly in the reproductive system, with reduction and alteration of the viteline follicles and the ovary of the females. Also modifications in the males testicles were observed. The results suggest that the drug under study reduces, considerably, the female S. mansoni, oviposition increases the worms' size, leads to alteration in the reproductive system of males and females. It also may induce death of a significant part of the worm populations at the dose of 400mg/kg.

A fim de investigar se a imunização de camundongos com a proteína recombinante de14,7 KDa (Sm14r) de Schistosoma mansoni confere proteção contra uma infecção letal por S. mansoni, camundongos imunizados com Sm14r foram infectados com diferentes cargas de vermes.Uma proteção significativa foi demonstrada nos camundongos imunizados e infectados com 100 ou 1.000 cercárias de S. mansoni quando comparados com os controles (p< 0,004 e p< 0,01 respectivamente). Diferentemente de resultados anteriores, nenhum camundongo do grupo controle (não imunizado e infectado com 1.000 cercárias) morreu antes do 30º dia após infecção. Uma correlação direta entre o número de cercárias e o tempo de morte dos camundongos foi detectada. Anticorpos IgM anti-Sm14r foram produzidos significativamente (p< 0,05), principalmente, nos grupos de camundongos imunizados e infectados com 500 ou 1.000 cercárias. Anticorpos IgG e IgA anti-Sm14r não foram produzidos em quantidades significativas. Os testes de immunoblots demonstraram que todos os soros de camundongos revelaram uma banda específica de aproximadamente 14,7 KDa, e com maior intensidade nos soros dos camundongos imunizados. A maior reatividade foi encontrada no soro no período da terceira imunização.Os resultados mostram que imunização com Sm14r reduz a carga de vermes independentemente da carga de cercárias da infecção. Nenhuma correlação foi encontrada entre tipos de anticorpos e redução da carga de vermes. Uma correlação direta foi encontrada em relação a carga de cercárias e a taxa e precocidade da mortalidade dos camundongos.

To investigate whether mice immunization with the recombinant form of a 14.7 KDa Schistosoma mansoni protein (rSm14) confers protection against a S. mansoni lethal challenge infection, rSm14-immunized mice were challenged with different cercarial burdens. A significant protection was detected in immunized mice challenged with 100 or 1,000 S. mansoni cercariae when compared with their controls (p< 0.004 and p< 0.01 respectively). Differently from previous report, none of the mice from the control group (not immunized and infected with 1000 cercariae) died before the 30th day post-infection. A direct correlation between the number of challenge cercariae and the precocity of mice death was found. IgM anti-rSm14 antibodies were significantly produced (p< 0.05) mainly in the groups of immunized mice infected with 500 or 1000 cercariae. IgG and IgA anti-rSm14 antibodies were not significantly detected. In Western immunoblots, all mice sera showed a specific antibody response with a 14.7 KDa antigen being reacted with particular intensity in sera from immunized mice. The results show that immunization with rSm14 reduced mice worm burden independently of the cercariae load of challenge infection. No correlation was found between serum antibodies and worm burden reduction. In relation to cercarial load and the rate and precocity of mice mortality a direct correlation was found.

Camundongos infectados experimentalmente com Schistosoma mansoni foram tratados, em dose oral única, com artesunato (LACTAB®), 300 ou 500mg/kg ou durante cinco dias consecutivos. Os animais foram sacrificados 7, 30, 60 ou 90 dias após o tratamento. Diferenças estatisticamente significativas foram encontradas na distribuição e mortalidade dos vermes e na alteração do oograma nos grupos tratados quando comparados ao controle, em todos os esquemas testados quando os animais foram sacrificados 30 dias após o tratamento. A análise morfológica dos vermes mostrou alterações no aparelho reprodutor feminino com diminuição do volume ovariano, rarefação dos folículos vitelínicos, alterações estas mais acentuadas nas dosagens mais altas, justificando o encontro na alteração do oograma que chegou a 100%. Entretanto, quando os animais foram sacrificados 60 ou 90 dias após o tratamento, as diferenças e alterações foram menores, mostrando que os vermes sobreviventes se recuperaram e reiniciaram a postura.

Mice experimentally infected with Schistosoma mansoni were treated orally with artesunate (Lactab®) in a single dose of 300 or 500mg/kg or over a period of five consecutive days. The animals were sacrificed 7, 30, 60 or 90 days after treatment. Statistically significant differences were found in the distribution and mortality of the worms and in the alterations of the oogram in the treated group when compared to control in all of the tested schemes when the animals were sacrificed 30 days after treatment. Morphological analysis of female worms showed a reduction of ovarian volume and rarefaction of the vitelline follicles. These modifications were more marked after treatment with the higher dose, explaining the alteration of the oogram which reached 100%. However, when the animals were sacrificed 60 or 90 days after treatment, the differences and alterations were smaller, showing that the surviving worms recovered and restarted oviposition.

Foram estudados dez isolados de Schistosoma mansoni provenientes de pacientes residentes em Itaquara, Bahia, Brasil, tratados com oxamniquine eposteriormente com praziquantel, e ainda assim não curados. Caramujos (Biomphalaria glabrataj foram expostos a miracídios provenientes das fezes dos pacientes. Cercãrías eliminadas por estes caramujos e por moluscos coletados no peridomicílio dos pacientes foram utilizadas para infecção experimental de camundongos albinos. Os animais infectados foram tratados em dose única, via oral, com oxamniquine (25, 50 e 100 mg/kg) ou praziquantel (100, 200 e 400mg/kg). Foram realizados estudos de análise e comparação de DNA de cercãrías de S. mansoni eliminadas pelos caramujos e de vermes adultos recolhidos de camundongos infectados experimentalmente com os isolados dos 10 pacientes e ainda de cercãrías de S. mansoni eliminadas por moluscos naturalmente infectados coletados em Itaquara. A cepa LE (mantida rotineiramente no laboratório) foi usada como padrão de comparação da resposta aos agentes esquistossomicidas administrados. As respostas terapêuticas foram significativamente diferentes entre alguns dos isolados embora não fosse possível caracterizar nenhum como resistente. A análise dos perfis de amplificação de DNA nas cercãrías e nos vermes adultos dos isolados de S. mansoni demonstrou baixo grau de variabilidade indicando que estes são geneticamente próximos e revelando a ausência de rearranjos globais dentro dos genomas.

Ten inhabitants of Itaquara, Bahia, Brazil treated with oxamniquine and subsequently praziquantel were not cured. Schistosoma mansoni isolates derived from these patients were studied. Snails were infected with miracidia derivedfrom the feces of these patients and the cercariae produced used to infect albino mice. The animals were then treated with a single oral dose of oxamniquine (25, 50 and 100mg/kg) or praziquantel (100, 200 and 400 mg/kg). Tloe response to chemotherapy was significantly different in some of the isolates although it was not possible to characterize any of them as resistant, hi addition, DNA analysis of the isolates by means of "Random Amplified Polymorfhic DNA ''indicated a low degree of variability as compared with a laboratory strain, LE. Thus, it was not possible to characterize these organisms at a genetic level as a distinct strain.

A densidade planorbídica, as taxas de infecção e a produção mensal de cercárias de Schistosoma mansoni por Biomphalaria glabrata, foram determinadas em foco do Barreiro de Baixo (Belo Horizonte MG, Brasil). Durante 38 meses, de 1984 a 1987, foram capturados 5.366 moluscos dentre os quais 324 (6,03%) estavam infectados com o S. mansoni. O total de cercárias eliminadas foi de 5.667.312. Cada molusco eliminou em média 17.422 cercárias durante sua permanência no laboratório. A maior longevidade dos moluscos infectados foi de 218 dias. Foi observada a cura natural de 42 (12,9%) dos exemplares infectados, após cerca de 130 dias de captura. A densidade planorbídica média, no foco durante esse período foi de 16,3 moluscos por conchada. A eliminação de cercárias pelos moluscos do campo foi comparada com a de exemplares criados no laboratório, infectados em massa com a linhagem LE de S. mansoni de Belo Horizonte. Os moluscos do laboratório eliminaram em média 6.061 cercárias por exemplar, número 2,8 vezes menor do que os moluscos do campo, devido a longevidade menor. A taxa de prevalência da esquistossomose no foco foi de 14,3%.

The snail density, levels of infection and the monthly production of Schistosoma mansoni cercariae by Biomphalaria glabrata were determined in a focus of Barreiro de Baixo (Belo Horizonte, MG, Brazil). During a period of 38 months (1984 to 1987) 5,366 snails were collected of which 324 (6.03%) were infected with S. mansoni. The total number of cercariae shed was 5,667,312. Each snail shed an average of 17,422 cercariae during the time that it was under study in the laboratory. The greatest longevity of infected snails was 218 days. Natural cure was observed in 42 (12.9%) of the infected specimens about 130 days after collection. The average snail density in the focus during the period of study was 16.3 snails per scoop. The shedding of cercariae by snails collected from the field was compared with laboratory bred specimens infected in mass with the LE strain of S. mansoni from Belo Horizonte. The laboratory infected snails shed an average of 6,061 cercariae each, a value 2.8 times less than the field specimens due to a shorter life span. The prevalence of schistosomiasis in the focus was 14.3%.

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

Aqueous suspensions of fruit's pericarp, leaves, root's bark and seeds of Guaiacum officinale were tested at different concentrations as molluscicide, cercaricide and piscicide. In the laboratory the suspension of fruit's pericarp produced 100% mortality for egg masses of B. glabrata at 100 ppm, for adult snails of Biomphalaria glabrata, B. straminea and B. tenagophila at 20 ppm, for Lebistes reticulatus (fishes) at 5 ppm and Schistosoma mansoni's cercariae at 1 ppm. The ethanolic extract of fruit's pericarp was not active against adult snails of B. glabrata. The letal dose for adult snails(DL90) of the aqueous suspension of fruit's pericarp after 24 hours exposure, were: 15 ppm for B. glabrata; 14 ppm for B. straminea and 18 ppm for B. tenagophila. The DL90 of aqueous suspensions of root's bark, seeds and leaves were 57 ppm, 33 ppm and 15 ppm, respectively. In the field, B. glabrata adult snail mortality was 68% at 20 ppm and 100% at 40 ppm, when using suspension of fruit's pericarp

The action of the ether artemisinin (artemether) on Shistosoma mansoni in mice and the hansters experimentally infected with the LE strain was studied. In mice, the drugs showed high schistosomicidal activity using a single intramuscular dose of 100 mg/Kg/day. By the oral route, this dose showed a low activity. Mice treated with a single intramuscular dose of 200 mg/Kg/day, and examined 15 days after treatment, presented 100% alteration of the oogram; when examined 45 days after treatment, the oogram was normal. With doses of 100 mg/Kg/day, i.m., during 3 or 5 consecutive days, the death rate of mice was very high. Morphologic analysis of the worms collected by perfusion of mice treated with a single dose of 100 mg/Kg/day, i.m., detected a marked decrease in the length of male and female forms, degenerative alterations in the parenchyma and in the reproductive system of the females, with reduction of vitellinic material and in ovary volume; the intestinal contents presented a marked despigmentation. In the male worms signifcant alteration was not apparent by optical microscopy.

Os hidrolatos e óleos essenciais de diferentes espécies de Eucalyptus foram testados sobre caramujos adultos e desovas de Biomphalaria glabrata e cercárias de Schistosoma mansoni. Esses produtos foram obtidos por arraste de vapor. Dos 21 hidrolatos ensaiados, oito foram ativos sobre caramujos, nove demonstraram ação sobre desovas e três apresentaram atividade sobre cercárias a partir da diluição de 1:4 (V/V). Dos óleos estudados, 11 tiveram atividade para planorbídeos e desovas nas concentrações de 20ppm (p/V). O hidrolato do Eucalyptus deanei foi ativo sobre caramujos, desovas e cercárias a partir da diluição de 1:4, e o óleo na concentração de 20 ppm.

Hydrolates and essential oils of several Eucalyptus species were tested on adult snails and egg masses of Biomphalaria glabrata, and cercaria of Shistosoma mansoni. These products were obtained by vapor dragging. Eight out of 21 hydrolates presented activity on snails, nine on egg masses and three on cercaria at 1:4 dilutions. Eleven oils were active on both snails and egg masses at 20 ppm concentrations. The hydrolate of E. deanei was active both on snails, egg masses and cercaria at 1:4 dilution and its essential oil at 20 ppm.

O extrato butílico da Phytolacca dodecandra (tipo 44) obtido de frutos procedentes da Etiópia, foi testado como moluscicida em nosso laboratório e no campo. As concentrações letais, CL90, com 24 horas de exposição, para Biomphalaria glabrata adulta, recém-eclodidas e desovadas foram de 4,5, 23,0 e 102,0 ppm, respectivamente. Para peixes, Lebistes reticulatus, a CL90 foi de 2,0 ppm. Esses resultados foram semelhantes aos obtidos por Lemma em 1984, na Etiópia. Em dois criadouros com água parada, tratados com 10ppm de estrato butílico e 3ppm de niclosamida, ocorreu 84,0 e 100,0% de mortalidade B. glabrata, respectivamente. Os dois produtos foram tóxicos para peixes (L. reticulatus) no campo. É discutida a possibilidade do uso de moluscidas de origem vegetal, como alternativa para o combate a focos de esquistossomose no Brasil.

A buthanol extract of Phytolacca dodecandra (type 44) obtained from Ethiopia berries, was tested as molluscicide in our laboratory and in the field. The lethal dose (LD90) for adult snails, newly hatched and egg-masses of Biomphalaria glabrata, in 24 hours exposure, were of 4.5, 23.0 and 102.0 ppm respectively. The LD90 for the fish Lebistes reticulatus was of 2.0 ppm. These results are similar to those of Lemma (1984) in Ethiopia. In two water ponds treated with 10 ppm of the buthanol extract or 3 ppm of niclosamide the mortality rates of B. glabrata were of 84.6 and 100.0%, respectively. Both treatments were toxic for L. reticulatus in the field trials. The possibility of using molluscicides derived from plants is discussed as an alternative for treatment of schistosomiasis foci in Brazil.

Caramujos Biomphalaria tenagophila descendentes de exemplares coletados no lago da Pampulha, Belo Horizonte, Minas Gerais, foram expostos a miracídios de quatro cepas de Schistosoma mansoni: LE e HK de origem local, Belo Horizonte, AL do Estado de Alagoas e SJ, de São José dos Campos, SP. As cepas LE, AL e SJ são mantidas em laboratório e HK foi obtida de fezes de paciente que reside próximo à Pampulha. As taxas de infecção experimental foram de 4% (LE), 6% (HK), 30% (SJ) e 40% (AL). Esses indícios de infecção foram semelhantes aos obtidos por vários autores para populações de B. tenagophila de Minas Gerais. Caramujos infectados experimentalmente eliminaram número de cercárias comparável ao de B. glabrata do controle e de B. tenagophila capturada no lago, com infecção natural (cerca de 2.000 cercárias/molusco). Devido à alta densidade planorbídica atual em alguns pontos do lago, número de cercárias eliminadas por exemplares naturalmente infectados, afluxo de pessoas para pesca e lazer, contaminação das águas por dejetos humanos, os autores alertam para o risco de crescimento do foco de esquistossomose no local.

Biomphalaria tenagophila snails, from a population originally obtained from "Pampulha" lake, Belo Horizonte, Minas Gerais State, Brazil, were exposed to miracidia from four strains of Schistosoma mansoni: "LE" and "HK" from Belo Horizonte, "AL" from alagoas and "SJ" from São José dos Campos, São Paulo. The "LE", "AL" and "SJ" strains are maintained in the laboratory and the "HK" strain was obtained from feces of a patient residing near to "Pampulha" lake. Infection rates were of 4% ("LE" strain), 6% ("HK" strain), 30% ("SJ" strain) and 40% ("AL" strain). These infection rates were similar to those obtained by others authors for B. tenagophila from Minas Gerais. Experimentally infected snails when compared to B. glabrata of the control group and B. tenagophila naturally infected in "Pampulha" lake shed similar number of cercariae (2000 cercariae/snail). The high density of B. tenagophila in the "Pampulha" lake, the number of cercariae shed by naturally infected snails, the great number of persons who use the water for fishing and swimming, and the water contamination with human feces, are favourable factors for growing the Schistosomiasis focus in the lake.