Treatment of acute experimental schistosomiasis

A model of acute schistosomiasis of the mouse was used to observe whether curative treatment would be followed by an enhancement of the hepatic and splenic lesions, as a consequence of the massive destruction of worms and eggs within the portal system. Mice infected with 50 cercariae of Schistosoma mansoni were treated with both oxamniquine and praziquantel on the 50th day of infection and submitted to a sequential histologic examination from the 2nd to the 45th day after treatment. Although severe focal lesions due to dead and disintegranting worms were present in the livers of the treated animals, no aggravation of the general changes (reative hepatitis and splenitis, or periovular granulomas) was seen in comparison with a control non-treated group. Of 50 animals treated during the acute phase of schistosomiasis only one died espontaneously, while 16 ou of 30 infected controls died before the end of the experiment. The present investigation indicates that curative treatment during the acute phase of schistosomiasis does not enhance previous lesions at first and results in progressive disappearance of the lesions starting six days following chemotherapy.

Resistance of schistosomes to hycanthone and oxamniquine

Genetic crosses between phenotypically resistant and sensitive schistosomes demonstrated that resistance to hycanthone and oxamniquine behaves like a recessive trait, thus suggesting that resistance is due to the lack of some factor. We hypothesized that, in order to kill schistosomes, hycanthone and oxamniquine need to be converted into an active metabolite by some parasite enzyme wich, if inactive, results in drug resistance. Esterification of the drugs seemed to be the most likely event as it would lead to the production of an alkylating agent upon dissociation of the ester. An artificial ester of hycanthone was indeed active even in resistant worms, thus indirectly supporting our hypothesis. In addition, several lines of evidence demonstrated that exposure to hycanthone and oxamniquine results in alkylation of worm macromolecules. Thus, radioactive drugs formed covalent bonds with the DNA of sensitive (but not of resistant) schistosomes; an antiserum raised against hycanthone detected the presence of the drug in the purified DNA fraction of sensitive (but not of resistant) schistosomes; a drug-DNA adduct was isolated from hycanthone-treated worms and fully characterized as hycanthone-deoxyguanosine.

Autoradiographic analysis of Schistosoma mansoni migration in the NZ rabbit

The migration of larval Schistosoma mansoni was tracked by means of autoradiographic analysis in naive rabbits percutaneously exposed to L-(**75 Se) selenomethionine-labeled cercariae on serial intervals of 1, 2, 4, 6, 8, 10, 15, 20, 25, 30, 40 and 50 days post-infection. Autoradiographic foci were detected from the 1st day in the skin, up to the 15th day in the liver. Adult and mature worms were recovered either paired or not 60 days after infection, by perfusion of hepatic and mesenteric veins. Morphometric analysis under optical microscopy, showed that worms were within regular dimention limits as compared to adult worms harboured by other host species. These observations extend previous informations on the S. mansoni-rabbit association and clearly demonstrate the post-liver phase of S.mansoni life-cycle in this host.

Identification and characterization of sex-linked proteins of Schistosoma mansoni

The proteins of adults worms (male and female) of two isolates (BH and RJ) of Shistosoma mansoni were extracted using Triton X-114 phase separation. The SDS-polyacrilamide gel electrophoresis profiles of the three phases (detergent, aqueous and insoluble proteins) obtained were compared after Coomassie blue and silver staining, surface radioiodination and Western blotting. No major differences were detected between the 2 isolates. Of the 25 or more proteins which partitioned into the detergent phase, only about 8 proteins could be surface radiodinated on live adult worms. A comparison was also made between the profiles of mael and females worms, isolated from bisexually infected mice. Two major female-specific and one male-specific band were detected by silver and/or Coomassie staining. The female bands, 32 KDa and 18 KDa, partitioned into the detergent and aqueous phase, respectively. The male-specific band of 42 KDa remained in the insoluble phase. Antigenic differences between male and females protins were detected by Western vlotting using a sera from infected Nectomys squamipes.

A new helminth parasite of fish: Spirocamallanus freitasi sp. n. (Nematoda - Camallanidae)

A new species of nematode is described, Spirocamallanus freitasi sp. n. The worms were collected from fishes (Bergiaria westermanni, Pimelodus maculatus and Pimelodus sp.) living in Três Marias Dam (São Francisco River) in the State of Minas Gerais, Brazil. Spirocamallanus freitasi sp. n. differs from Procamallanus iheringi, P. amarali, P. macaensis, P. (Spirocamallanus) pimelodus, P. (S.) solani and P. (S.) pereirai by having digitated larger spicule and from P. cruzi by having not digitated terminal parts of both spicules. It differs also from P. (S.) intermedius by having not larger spicule divided in two undigitated branches and six to nine sclerotized bands in the buccal capsule; P. rarus has three to four sclerotized bands and tridigitated larger branch of longer speicule, differing from S. feitasi sp. n.

Travassosnema gen. n. with the description of T. travassosi sp. n. (Dracunculoidea, Guyanemidae) parasite of Acestrorhynchus lacustris Reinhardt, 1874 (Characidae) from Três Marias Reservoir, MG, Brazil

A new genus, Travassosnema (Guyanemidae, Dracunculoidea) is proposed to include filariid worms having esophagus divided into muscular and glandular parts, with esophageal appendix near junction with intestine; anus functional; vulva anterior, well developed and functional in mature females. Travassonema travassosi sp. n., a parasite of Acestrorhynchus lacustris Reinhardt, 1874 from Três Marias Reservoir (São Francisco River) in the State of Minas Gerais, Brazil, is described. The generic and the specific names are a tribure to Brazilian parasitologist Lauro Travassos at his birth centenary.

Activity of the artemether in experimental schistosomiasis mansoni

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.

Parasites of commercial shrimps and fishes in Argentine sea: on the adult and metacercaria of Opecoeloides feliciae n. sp. (Digenea: Opecoelidae)

Opecoeloides feliciae n. sp., first record of opecoelid metacercariae in commercial shrimps of South Atlantic Ocean, parasitizing Cynoscion striatus (adults) and Artemesia longinaris (metacercariae), is described, illustrated and compared with related species of the genus. Adults, immature worms and metacercariae are compared, and rates of prevalence and intensity of infection are also given.

Experimental heteroxenous cycle of Lagochilascaris minor Leiper, 1909 (Nematoda: Ascarididae) in white mice and in cats

Reports of natural infections of sylvatic carnivores by adult worms of species similar to Lagochilascaris minor in the Neotropical region led to attempts to estabilish experimental cycles in laboratory mice and in cats. Also, larval development was seen in the skeletal muscle of an agouti (Dasyprocta leporina) infected per os with incubated eggs of the parasite obtained from a human case. In cats, adult worms develop and fertile eggs are expelled in the feces: in mice, larval stages of the parasite develop, and are encapsulate in the skeletal muscle, and in the adipose and subcutaneous connective tissue. From our observations, we conclude that the larva infective for the mouse is the early 3rd stage, while for the final host the infective form is the later 3rd stage. A single moult was seen in the mouse, giving rise to a small population of 4th stage larvae, long after the initial infection.

Ultrastructure of the adults of Bunostomum phlebotomum (Nematoda: Ancylostomatidae)

The morphology of the male and female of Bunostomum phlebotomum are described based on scanning electron microscope (SEM) observations. The attachment of the worms to the small intestinal mucosa and during the copula were observed. Structures of the bucal capsule and genital organs were also studied.

Natural infection of wild rodents by Schistosoma mansoni parasitological aspects

The evaluation of the role of rodents as natural hosts of Schistosoma mansoni was studied at the Pamparrão Valley, Sumidouro, RJ, with monthly captures and examination of the animals. Twenty-three Nectomys squamipes and 9 Akodom arviculoides with a shistosomal infection rate of 56.5% and 22.2% respectively eliminated a great majority of viable eggs. With a strain isolated from one of the naturally infected N. squamipes, we infected 75% of simpatric Biomphalaria glabrata and 100% of albino Mus musculus mice. The adult worms, isolated from N. squamipes after perfusion were located mainly in the liver (91.5%) and the mesenteric veins (8.5%). The male/female proportion was 2:1. The eggs were distributed on small intestine segments (proximal, medial and distal portions) and the large intestine without any significant differences in egg concentration of these segments. In A. arviculoides, the few eggs eliminated by the stools were viable and there was litlle egg retention on intestinal segments. Considering the ease to complete S. mansoni biological cycle in the Nectomys/Biomphalaria/Nectomys system under laboratory conditions, probably the same is likely to occur in natural conditions. In support to this hypotesis there are also the facts that human mansonic shistosomiasis has a very low prevalence in Sumidouro and endemicity among the rodents has not changed even after repetead treatments of the local patients. Based on our experiments, we conclude that N. squamipes has become a natural host of S. mansoni and possibly may participate in keeping the cycle of schistosomiasis transmission at Pamparrão Valley.

Nectomys squamipes (Rodentia: Cricetidae) as an experimental model for schistosomiasis mansoni

Twenty specimens of Nectomys squamipes born in captivity, were infected with 500 cercariae by the transcutaneous route. Coprologic examinations were carried out from the 5th to 23rd week after infection. On the 7th, 8th, 12th, 16th, and 23rd weeks the animals were sacrificed and perfused. The oogram was performed in segments of the small intestine (proximal, medial and distal portions) and the large intestine. The average pre-patent period was of 42 days. The average number of eggs varied from 350 on 6th week, to 800 on the 13th. From the 14th week on, the average number of eggs eliminated was lower than 50 per gram of feces. The recovery of worms kept steady on the 7th, 8th, and 12th week (16.85%; 15.45% and 11.95%), decreasing to 7.70% on the 16th week and 8.45% on the 23rd week. The proportion of male/female worms was about the same on the first two weeks, but from the 12th week on, the proportion was: 1,4/1 on the 12th week; 2,5/1 on the 16thweek and 1,8/1 on the 23rd weekThese observations suggest that N. squamipes may used as an experimental model for schistosomiasis mansoni, to wich it develops resistance mechanism, useful for immunity studies.

Parasite enzymes as a tool to investigate immune responses

Previous evidences reported by us and by other authors revealed the presence of IgG in sera of Schistosoma mansoni-infected patients to immunodominant antigens which are enzymes. Besides their immunological interest as possible inductors of protection, several of these enzume antigens might be also intersting markers of infection in antibody-detecting immunocapture assays which use the intrinsic catalytic property of these antigens. It was thus thought important to define some enzymatic and immunological characteristics of these molecules to better exploit their use as antigens. Four different enzymes from adult worms were partially characterized in their biochemical properties and susceptibility to react with antibodies of infected patients, namely alkaline phosphatase (AKP, Mg*+, pH 9.5), type I phosphodiesterase (PDE, pH 9.5), cysteine proteinase (CP, dithiothreitol, pH 5.5) and N-acetyl-ß-D-glucosaminidase (NAG, pH 5.5). The AKP and PDE are distinct tegumental membrane-bound enzymes whereas CP and NAG are soluble acid enzymes. Antibodies in infected human sera differed in their capacity to react with and to inhibit these enzyme antigens. Possibly, the specificity of the antibodies related to the extent of homology between the parasite and the host enzyme might be in part responsible for the above differences. The results are also discussed in view of the possible functional importance of these enzymes.

Human IgE responses to Schistosoma mansoni and resistance to reinfection

Schistosoma mansoni infected Kenyan patients were treated and the intensities of their reinfections were followed over the next two years. in addition, their pre- and six month post-treatment serum levels of IgG1-4, IgM, and IgE, specific for schistosoma, egg and adult worm, were measured in ELISA. No reinfection took place before six months post-treatment. Reinfection intensities varied with age; the younger children becoming reinfected at significantly higher intensities than older individuals. When antibody and reinfection levels were compared, only the six month post-treatment IgE response against adult worm correlated negatively with intensities of reinfection and, therefore, was predictive of resistance or immunity to reinfection. IgE and IgG specific Western Blots were carried out. The adult worm antigens recognized by IgE were restricted compared with the IgG responses of the same patients, although no individual antigen was uniquely recognized by the IgE isotype. A dominant 22 kDa antigen was recognized by most but not all high IgE responders. Patients with IgE responses against this antigen suffered significantly lower subsequent levels of reinfection, compared with non-responders. A monospecific rabbit antiserum against the 22KDa adult worm antigen showed that this antigen is specifically located in the tegument of the adult worm and of 'lung' and 'liver' stage schistosomula, but is absent from the early 'skin' schistosomula. It is possible that this antigen is a target for human IgE mediated immune effector mechanisms active against the post skin stage schistosomula and that this is boosted by the death of adult worms.

Immunological profiles of patients from endemic areas infected with Schistosoma mansoni

Crude extracts of eggs (SEA) adult worms (SWAP) or cercariae (Cerc) have been used to stimulate Peripheral Blood Mononuclear cells (PBMC) and have provided rather distinct profiles of responses in different types of patients. In genenral it is clear that patients with early infections respond strongly to SEA while response to SWAP are developed more slowly. As infection progresses into the more chronic phases, a general pattern is seen whic leads to lower anti-SEA proliferative responses in the face of higher responses to SWAP and variable anti-cerc responsiveness. Cured not re-exposed patients express very high levels of anti-SEA proliferation. It has recently been seen that those individuals who live in endemic areas and have continued water contact, but are reapeatedly stool-negative (who are presumed to have self-cured or be putatively resistant; endemic normals) are strongly responsive to antigenic extracts, particularly to SEA. Furthermore, our results show that endemic normal individuals have significantly higher IFN gamma production upon PBMC stimulation with schistosome antigens than infected individuals. With the emergence of more studies it is becoming apparent that both the intensity and the prevalence of a given area may influence or shape the general responsiveness of the population under study.