Intermediate hosts of schistosoma mansoni in Brazil

The Brazilian planorbidical chart is slowly but progressively been increased by new data. Distribution of vector species of Schistosoma mansoni, according to Paraense, 1986, may be thus resumed: Biomphalaria glabrata - delimited by paralells 13 and 21-S and meridians 39 and 45-W, area of greater dominance (Southerst Bahia, oriental half of Minas Gerais and Espírito Santo). It is observed along the coast line of the state of Sergipe, Alagoas, Pernambuco, Paraíba and Rio Grande do Norte. Starting from there, it is found towards the southwest, in the direction to the Sao Francisco River and South-Center of Minas Gerais. Isolated population may be observed in other states. Its presence is probably, associated to the transmission of schistosomiasis in all areas where it occurs. B. tenagophila - extends it self through a wide strip of coast-line the South of Bahia (17-45"S, 39-15'W), RS(33-41'S, 53-27'W). In Sao Paulo and Rio Grande do Sul states it is found further inland. It is important in schistosomiasis transmission in the Paraíba valley (SP). Isolated populations are observed in the Federal District and Minas Gerais state. B. straminea - better adapter species to climatic variation, having a more dense ditribution in the northeast (41-Wand 110-S), south of Bahia and northeast of Minas Gerais (150 and 180-S, 400 and 440-W) It is less susceptible than B. glabrata, being however the most important responsible for the transmission of S. mansoni in the northeast, chiefly in the northeastern dry area, where it is almost the only transmissive species.

On the diversity of mollusc intermediate hosts of Angiostrongylus costaricensis Morera & Cespedes, 1971 in southern Brazil

Veronicellid slugs are considered the most important intermediate hosts of Angiostrongylus costaricensis, an intra-arterial nematode of rodents. Studies undertaken in three localities in southern Brazil led to identification of molluscs other than veronicellid slugs as hosts of A. costaricensis: Limax maximus, Limax flavus and Bradybaena similaris. These data indicate a low host specificity of larval stages of A. costaricensis, as it has been reported to other congeneric species.

Eurytrema coelomaticum (Digenea, Dicrocoeliidae): the effect of infection on carbohydrate contents of its intermediate snail host, Bradybaena similaris (Gastropoda, Xanthonychidae)

The interface Eurytrema coelomaticum/Bradybaena similaris was studied by quantifying the amount of glucose on the hemolymph and the content of glycogen in the cells of the digestive gland and the cephalopedal mass of infected and uninfected snails. Samples were analyzed on days 0, 30, 90 and 150 post-infection. The infected snails had less glucose in the hemolymph, with a reduction of 67.05 por cento at 30 days, and 62.09 por cento at 90 days post-infection. The reduction in glycogen content was 86.41 por cento in the digestive gland and 79.1 por cento in the cephalopedal mass at 30 days, and 92.71 por cento and 90.89 por cento in these organs respectively at 90 days post-infection. It is proposed that the sporocysts absorb glucose directly from the hemolymph.

Degree of host-parasite compatibility between Schistosoma mansoni and their intermediate molluscan hosts in Brazil

The compatibility of Biomphalaria tenagophila, B. straminea and B. glabrata from Minas Gerais with different strains of Schistosoma mansoni was evaluated using the method of Frandsen (1979b) in standardized experiments. One hundred and fifty of each species of snail were individually exposed in the laboratory to 50 miracidia of S. mansoni lines LE, SJ and AL. The cercariae from the infected snails were counted and used to calculate TCP/100 indices, which were compared with those of Frandsen (1979b). For B. tenagophila the TCP/100 indices varied from 37,996 to 74,266 (class II and III). The snail was poorly compatible with LE (class II) and compatible with SJ and AL (class III). For B. straminea the indices varied from 9,484 to 20,508. The snail was not very compatible with SJ (class I) and poorly compatible with LE and AL (class II). For B. glabrata the indices varied from 588,828 to 1,039,065. The snails was extremely compatible (class VI) with the three lines of S. mansoni. These results confirm the epidemiological importance of B. glabrata in Brazil followed by B. tenagophila and B. straminea.

Transfer of toxin genes to alternate bacterial hosts for mosquito control

Mosquitoes are vector of serious human and animal diseases, such as malaria, dengue, yellow fever, among others. The use of biological control agents has provide an environmentally safe and highly specific alternative to the use of chemical insecticides in the control of vector borne diseases. Bacillus thuringiensis and B. sphaericus produce toxic proteins to mosquito larvae. Great progress has been made on the biochemical and molecular characterization of such proteins and the genes encoding them. Nevertheless, the low residuality of these biological insecticides is one of the major drawbacks. This article present some interesting aspects of the mosquito larvae feeding habits and review the attempts that have been made to genetically engineer microorganisms that while are used by mosquito larvae as a food source should express the Bacillus toxin genes in order to improve the residuality and stability in the mosquito breeding ponds.

A low stringency polymerase chain reaction approach to the identification of Biomphalaria glabrata and B. tenagophila, intermediate snail hosts of Schistosoma mansoni in Brazil

The low stringency-polymerase chain reaction (LS-PCR) with a pair of specific primers for the amplification of the 18S rRNA gene was evaluated as a means of differentiating between the two Schistosoma mansoni intermediate host species in Brazil: Biomphalaria glabrata and B. tenagophila. Individual snails obtained from different states of Brazil were used and the amplification patterns obtained showed a high degree of genetic variability in these species. Nevertheless, 4 and 3 clearly defined specific diagnostic bands was observed in individuals from B. glabrata and B. tenagophila respectively. The detection of snail specific diagnostic bands suggests the possibility of reliable species differentiation at the DNA level using LS-PCR.

Abdominal Angiostrongylosis in Southern Brazil - Prevalence and Parasitic Burden in Mollusc Intermediate Hosts from Eighteen Endemic Foci

Angiostrongylus costaricensis is a parasitic nematode of rodents and molluscs are the intermediate hosts. Nocturnal collection of molluscs and search for infective third stage larvae of A. costaricensis was carried out in 18 endemic foci identified by the notification of a confirmed diagnosis in human biopsies or surgical specimens. Molluscs were digested in acidic solution and isolation of larvae eventually present was done in a Baermann funnel. Larvae identified by the presence of a delicate groove in the tail were counted to assess the individual parasitic burden. Four species were found infected, with ranges of prevalence in parenthesis: Phyllocaulis variegatus (7% to 33.3%); Bradybaena similaris (11.7% to 24.1%); Belocaulus angustipes (8.3% ) and Phyllocaulis soleiformis (3.3% to 14.2%). Parasitic burden varied from 1 to 75 with P. variegatus, 1 to 98 with B. similaris, 1 to 13 with B. angustipes and 1 larvae in each of two specimens of P. soleiformis. P. variegatus was present in all sites and was found infected with the highest prevalence figures and the highest individual parasitic burdens. These data stress the importance of veronicellid slugs as intermediate hosts for A. costaricensis in the endemic areas in Rio Grande do Sul, Brazil

Genetic and Antigenic Analysis of Adenovirus Type 3 Strains Showing Intermediate Behavior in Standard Seroneutralization Test

During an epidemiological survey of acute respiratory infection in Rio de Janeiro, among 208 adenovirus isolates, we found two strains that we were not able, by a standard neutralization procedure, to distinguish between type 3 or 7. However, DNA restriction pattern for the two strains with different enzymes were analyzed and showed a typical Ad3h profile. Using a cross-neutralization test in which both Ad3p and Ad7p antisera were used in different concentration against 100 TCID50 of each adenovirus standard and both isolates, we were able to confirm that the two isolates belong to serotype 3. An hemagglutination inhibition test also corroborated the identification of both strains as adenovirus type 3. Comparing Ad3h and Ad3p genome, we observed 16 different restriction enzyme sites, three of which were located in genomic regions encoding polypeptides involved in neutralization sites

Genetic variability and identification of the intermediate snail hosts of Schistosoma mansoni

Studies based on shell or reproductive organ morphology and genetic considerations suggest extensive intraspecific variation in Biomphalaria snails. The high variability at the morphological and genetic levels, as well as the small size of some specimens and similarities between species complicate the correct identification of these snails. Here we review our work using methods based on polymerase chain reaction (PCR) amplification for analysis of genetic variation and identification of Biomphalaria snails from Brazil, Argentina, Uruguay and Paraguay. Arbitrarily primed-PCR revealed that the genome of B. glabrata exihibits a remarkable degree of intraespecific polymorphism. Low stringency-PCR using primers for 18S rRNA permited the identification of B. glabrata, B. tenagophila and B. occidentalis. The study of individuals obtained from geographically distinct populations exhibits significant intraspecific DNA polymorphism, however specimens from the same species, exhibit some species specific LSPs. We also showed that PCR-restriction fragment of length polymorphism of the internal transcribed spacer region of Biomphalaria rDNA, using DdeI permits the differentiation of the three intermediate hosts of Schistosoma mansoni. The molecular biological techniques used in our studies are very useful for the generation of new knowledge concerning the systematics and population genetics of Biomphalaria snails.

Molecular characterisation of intermediate snail hosts and the search for resistance genes

The relationship between schistosomes and their intermediate hosts is an extremely intricate one with strains and species of the parasite depending on particular species of snail, which in turn may vary in their susceptibility to the parasites. In order to gain a better understanding of the epidemiology of the disease we have been investigating the use of molecular markers for snail identification and for studying host-parasite relationships. In this paper we will draw on examples concerning schistosomiasis in West and East Africa to illustrate how a molecular analysis can be used as part of a "total evidence" approach to characterisation of Bulinus species and provide insights into parasite transmission. Particular emphasis is given to ribosomal RNA genes (rRNA), random amplified polymorphic DNA (RAPDs) and the mitochondrial gene cytochrome oxidase I (COI). Snails resistant to infection occur naturally and there is a genetic basis for this resistance. In Biomphalaria glabrata resistance to Schistosoma mansoni is known to be a polygenic trait and we have initiated a preliminary search for snail genomic regions linked to, or involved in, resistance by using a RAPD based approach in conjunction with progeny pooling methods. We are currently characterising a variety of STSs (sequence tagged sites) associated with resistance. These can be used for local linkage and interval mapping to define genomic regions associated with the resistance trait. The development of such markers into simple dot-blot or specific PCR-based assays may have a direct and practical application for the identification of resistant snails in natural populations.

Wild Rodents as Experimental Intermediate Hosts of Lagochilascaris minor Leiper, 1909

A total of 25 specimens of Cavia porcellus (guinea pig), 5 Dasyprocta agouti (agouti), and 22 Calomys callosus (vesper mice) were inoculated with infective eggs of Lagochilascaris minor. The inoculum was prepared with embryonated eggs and orally administered to each individual animal through an esophagus probe. In parallel, 100 specimens of Felis catus domesticus were individually fed with 55-70 nodules containing 3rd-stage larvae encysted in tissues of infected rodents. Animals were examined and necropsied at different time intervals. The migration and encystment of L3 larva was observed in viscera, skeletal muscle, adipose and subcutaneous tissues from all rodents. Adult worms localized at abscesses in the cervical region, rhino, and oropharynx were recovered from domestic cats inoculated with infected rodent tissues. Through this study we can conclude that: (1) wild rodents act as intermediate hosts, characterizing this ascarid heteroxenic cycle; (2) in natural conditions rodents could possibly act as either intermediate hosts or paratenic hosts of Lagochilascaris minor; (3) despite the occurrence of an auto-infecting cycle, in prime-infection of felines (definite hosts) the cycle is only completed when intermediate hosts are provided; and (4) in the wild, rodents could serve as a source of infection for humans as they are frequently used as food in regions with the highest incidence of human lagochilascariasis.

Penetration Sites and Migratory Routes of Angiostrongylus costaricensis in the Experimental Intermediate Host (Sarasinula marginata)

The intermediate hosts of Angiostrongylus costaricensis are terrestrian molluscs, mostly of the family Veronicellidae. The present work aimed at clarifying more accurately the sites of penetration and the migratory routes of A. costaricensis in the tissue slugs and at verifying the pattern of the perilarval reaction at different times of infection. Slugs were individually infected with 5,000 L1, and killed from 30 min to 30 days after infection. From 30 min up to 2 hr after infection, L1 were found within the lumen of different segments of the digestive tube having their number diminished in more advanced times after exposition until complete disappearance. After 30 min of exposition, percutaneous infection occurred, simultaneously to oral infection. Perilarval reaction was observed from 2 hr of infection around larvae in fibromuscular layer, appearing later (after 6 hr) around larvae located in the viscera. A pre-granulomatous reaction was characterized by gradative concentration of amebocytes around larvae, evolving two well-organized granulomas. In this work we confirmed the simultaneous occurrence of oral and percutaneous infections. Perilarval reaction, when very well developed, defined typical granulomatous structure, including epithelioid cell transformation. The infection also caused a systemic mobilization of amebocytes and provoked amebocyte-endothelium interactions.