Apoptotic mechanisms are involved in the death of Strongyloides venezuelensis after triggering of nitric oxide.

Despite progress in understanding the role of nitric oxide (NO) in the pathogenesis of helminth infections, the role in strongyloidosis is unknown. Firstly, we studied the production of NO in mice infected with Strongyloides venezuelensis as well as in macrophage cultures stimulated with parasite antigens. Somatic larvae 3 (L3) and excretory-secretory female antigens stimulate specific NO production measured by Griess reaction and expression of inducible NO synthase by RT-PCR and quantitative PCR. Moreover, mice infected with S. venezuelensis produce NO in migration stages. Secondly, we analysed the effect of NO production on L3 and females of S. venezuelensis using NO donors such as diethylenetriamine and 3,3-bis(aminoethyl)-1-hydroxy-2-oxo-1-triazene. Parasites died after NO donor treatment in a dose-dependent manner. Finally, apoptotic mechanisms are involved in the death of S. venezuelensis larvae.

Development of Fasciola hepatica in Lymnaea columella infected with miracidia derived from cattle and marmoset infections.

The development of Fasciola hepatica from two species of definitive hosts, i.e. cattle (Bos taurus) and a marmoset (Callithrix penicillata) in the snail Lymnaea columella was determined based on the production of rediae and cercariae and snail survival rate. More rediae and cercariae at 60-74 days post-infection were produced by snails infected by cattle-derived miracidia (cattle group) than by those infected by marmoset-derived miracidia (marmoset group). Among the L. columella parasitized by the marmoset group, the survival rate and the percentage of positive snails were higher than among those parasitized by the cattle group. Eggs of F. hepatica released in cattle faeces were significantly bigger than those released in marmoset faeces. Miracidia originating from parasites that completed their development in cattle were more efficient in infecting the intermediate host. These results suggest that vertebrate-host origin influences the eggs produced by the parasite and the infection rates in the snail host L. columella.

Evaluation of the schistosomicidal efficacy of liposome-entrapped oxamniquine.

Oxamniquine (OXA) was successfully encapsulated in small unilamellar vesicles using a pH gradient method. This procedure led to a high drug encapsulation efficiency (> 85%) at a drug to lipid molar ratio of 1/10. Moreover, these liposomes were found to retain encapsulated OXA efficiently under dialysis conditions at 37 degrees C. Liposome-entrapped OXA (LOXA), OXA, and empty liposomes were tested against Schistosoma mansoni in a murine model. LOXA produced a significant reduction of the worm burden compared to the other preparations, when inoculated by subcutaneous route (s.c.) with 10 mg OXA/kg animal one day before the infection, and 3, 7, and 14 days after. However, LOXA was not effective when given 7 days before, or 35 days after infections. OXA, in the free form, was effective in relation to the untreated group, only when administered 3 days after the infection. Maximum effect of LOXA, with 97% reduction of the parasite number, was observed when the preparation was given s.c. one day before the infection. On the other hand, LOXA inoculated intraperitoneally one day before the infection didn't show any reduction of the parasite count. It can be concluded that LOXA is more effective than OXA for the treatment of experimental schistosomiasis, particularly when administered subcutaneously at a time close to the infection.

Effects of ivermectin on Culex quinquefasciatus larvae.

The effects of ivermectin, a semi-synthetic drug widely used for treatment of livestock parasite diseases, were observed on Culex quinquefasciatus larvae. Toxic effects and mortality evaluation were carried out after 5, 15, 30 and 60 minutes of exposure to 1, 5 or 10 ppm of ivermectin solutions. Observations were made 24 and 48 hours after the beginning of the experiment, and loss of mobility, progressive paralysis and high mortality of larvae were recorded. The observed effects of ivermectin on the mosquito larvae is probably correlated with chloride channel activation on cell membranes.

[Susceptibility of Biomphalaria tenagophila and Biomphalaria glabrata from a same region to 2 Schistosoma mansoni strains]. / Suscetibilidade de Biomphalaria tenagophila e Biomphalaria glabrata de uma mesma região a duas cepas de Schistosoma mansoni.

B. tenagophila snails from Ouro Branco, MG, showed positivity for S. mansoni, with infection rates of 5%, 10%, (SJ strain), and 1% (LE strain) using a pool of miracidia. The mollusks were found to be susceptive from the 3rd generation reared in laboratory onwards. The B. tenagophila (OB, MG) when individually exposed to 10 miracidia, showed infection rate of 2% for LE strain. B. glabrata snails from Gagé, MG, showed a positivity rate of 58% for S. mansoni (LE strain), under experimental conditions. The B. tenagophila from Cabo Frio, RJ and B. glabrata from Belo Horizonte, MG used as a control for SJ strain showed infection rates of 47%-85% and 36% respectivily. For the LE strain, B. glabrata (BH, MG) used as control showed infection rate of 40%-75%.

[Malacological survey of the Soledade lake hydrographic basin, in Ouro Branco (Minas Gerais, Brazil)]. / Levantamento malacológico da bacia hidrográfica do Lago Soledade, Ouro Branco (Minas Gerais, Brasil).

A malacological survey was carried out at the Soledade Lake, in Ouro Branco, State of Minas Gerais, for the period 1986-1991. A total amount of 46,579 mollusks was collected, and among them seven species corresponding to five families could be found, as follows: 39,176 specimens of Biomphalaria tenagophila; 1,296 B. glabrata; 7 Drepanotrema cimex; 2,527 Physa sp; 417 Lymnaea sp; 92 Pomacea hastrum, and 3,064 specimens of Melanoides tuberculata (Melanniidae = Thiaridae) were collected from March/1990 onwards. Four specimens of B. tenagophila were found to be positive for Schistosoma mansoni.

Oogram studies in mice infected with Schistosoma mansoni and treated with dexamethasone.

Mice infected with about 90 cercariae of Schistosoma mansoni (LE strain) were treated during five consecutive days with dexamethasone (50 mg/Kg, subcutaneously), starting on the 42nd day of infection. Groups of five mice were then daily sacrificed from the first day after onset of treatment until the first day after. The perfusion of the portal system was performed and a piece of the intestine was processed for qualitative and quantitative oograms. This treatment carries to larger numbers of eggs in the tissues of treated mice, when compared with untreated groups. No changes were observed in the kinetics of oviposition, as all stages of viable eggs were observed in the tissues of treated and control mice. These data reinforce the hypothesis of a partial blockade of the egg excretion in immunosuppressed mice.

Schistosoma mansoni: the effect of dexamethasone on the cercaria-schistosomulum transformation, in vivo.

Treatment with dexamethasone (DMS) in the early phases of the experimental Schistosoma mansoni infection causes an indirect effect on the cercaria-schistosomulum transformation process. This is observed when naive albino mice are treated with that drug (50 mg/Kg, subcutaneously) and infected intraperitoneally 01 hour later with about 500 S. mansoni cercariae (LE strain). An inhibition in the host cell adhesion to the larvae, with a simultaneous delay in the cercaria-schistosomulum transformation, is observed. This effect is probably due to a blockade of the neutrophil migration to the peritoneal cavity of mice, by an impairment of the release of chemotactic substances. Such delay probably favors the killing of S. mansoni larvae, still in the transformation process, by the vertebrate host defenses, as the complement system.

[Schistosoma mansoni development in the peritoneal cavity of inbred AKR/J mice]. / Desenvolvimento do Schistosoma mansoni na cavidade peritoneal de camundongos da linhagem AKR/J.

Cercariae of Schistosoma mansoni after intraperitoneal inoculation, developed and reached the sexual maturation in that site with egg production. In the AKR/J strain of mice, 7.7% of the peritoneal recovered females showed normal eggs in the uterus. No evidence of hemoglobinic pigment in their digestive tract was observed in the peritoneal recovered parasites from both strains (AKR/J and SWISS). This fact suggests that the parasites can develop without red blood cells ingestion. On the other hand, the development of the parasite with egg production in the peritoneal cavity of the AKR/J mouse reinforces the data that the lung phase is not necessary for the development of the parasite.

Host cell adhesion to Schistosoma mansoni larvae in the peritoneal cavity of naive mice. Histological and scanning electron microscopic studies.

Cercariae of Schistosoma mansoni inoculated into the peritoneal cavity of naive mice induced host cell adhesion to their surface, but after 90 minutes the number of adherent cells sharply decreased. The cell detachment is progressive and simultaneous to the cercaria-schistosomule transformation. The histological study showed mainly neutrophils in close contact with the larvae. Mononuclear cells and some eosinophils were occasionally seen surrounding the adherent neutrophils. The scanning electron microscopy showed cells displaying twisted microvilli and several microplicae contacting or spreading over the larval surface, and larvae completely surrounded by clusters of cells. These results suggest that the neutrophils recognize molecules on the cercarial surface which induce their spreading.

Interaction between neutrophils and Schistosoma mansoni larvae in vivo. A transmission electron-microscopic study.

Schistosoma mansoni cercariae were inoculated into the peritoneal cavity of naive mice and recovered 30 minutes later. Ultrastructural studies showed that neutrophils adhere to the larval surface and participate in the removal of glycocalyx by phagocytosis. This finding suggests that the neutrophils can play a role on the cercaria-schistosomulum transformation process.

Kinetics of the pulmonary phase of Schistosoma mansoni in mice treated with dexamethasone.

In the experimental schistosomiasis mansoni glucocorticoids cause a reduction in the worm burden when administered in the week of infection or, the longest, at the next week. In order to determinate the probable(s) site(s) of reduction of the worm burden, mice were infected with cercariae of LE strain of S. mansoni and dexamethasone was administered daily (50 mg/kg, subcutaneously) starting 1 hour before infection until the eighth day. Mice were sacrificed daily starting on the third day after infection until the ninth day, and schistosomula from lungs were collected. Six weeks after infection, the remaining mice were sacrificed and perfused for adult worm recovery. Analysis of the results showed that the non-treated mice presented larger numbers of lung larvae than the treated ones, and this difference was also found later in the worm burden in the portal system. This difference may reflect the early death of larvae in treated animals, before or after reaching the lungs.