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Susceptibility of South African Bulinus africanus populations to infection with Schistosoma haematobium.

The use of intermediate host snails of Schistosoma spp. which are insusceptible to infection has been proposed as a possible method of controlling schistosomiasis. The objective of this approach is to change the susceptibility of natural snail populations from being predominantly susceptible to a non-susceptible state, through the release of refractory snails into natural habitats. In an attempt to determine whether or not such refractory Bulinus africanus populations occur in eastern South Africa, F1 generation snails of populations from 8 different areas were exposed to miracidia hatched from eggs excreted in the urine of schoolchildren infected with Schistosoma haematobium in the Nelspruit district. The proportion of snails successfully infected ranged from 27 to 100%, which revealed considerable genetic heterogeneity amongst populations of the same snail species. One population from Natal could be regarded as partially refractory, while a laboratory population from Durban proved to be 100% susceptible. A completely refractory strain of B. africanus has not yet been identified.

Animals↗

The effects of starvation and refeeding on egg laying and the synthetic activity of the albumen gland in Bulinus truncatus, a snail vector of urinary schistosomiasis.

1. The effects of starvation (for 1, 2, 3, 6, 9 and 12 days, respectively) and refeeding (12 days starvation and 1, 2 and 3 days refeeding, respectively) on egg laying and albumen gland activity in the freshwater snail, Bulinus truncatus were studied. 2. The egg laying of starving snails rapidly decreased and ceased by day 6 of starvation. Egg laying was restored 24 hr after refeeding. The recorded decrease in albumen gland wet weight was proportional to the starvation periods. The DNA contents of the glands of the different experimental groups was not statistically different from the controls. 3. Albumen gland synthetic activity expressed as 14C-glucose incorporation into galactogen/microgram DNA and 3H-amino acids into total protein was determined. The glands showed an abrupt decrease in synthetic activity after 1 day of starvation and gradually decreased further until days 9-12. The decrease in activity of the glands was more rapid than that of egg laying. Upon refeeding, the activity of the glands recovered rapidly, simultaneous with the increase in wet weight and egg laying. 4. In conclusion, there is a correlation between egg laying and the in vitro activity of albumen glands. The results show a short-term effect of starvation on the fecundity of the snails. Such studies could be useful in field studies as well as snail control by applying molluscicides under optimal conditions.

Animals↗

The carboxylic and amino acid chemoreception niche of Bulinus rohlfsi (Clessin), the snail host of Schistosoma haematobium.

The response of the freshwater snail, Bulinus rohlfsi, to gradients of carboxylic acids, amino acids and some related compounds was investigated by means of diffusion olfactometers. The snails proved to be very discriminating in their responses to the chemical species tested and there were also very marked ontogenetic differences. Although 15 distinctive types of amino and carboxylic acids proved to be attractants and arrestants, only octanoic acid proved effective for snails of all ages. Possible mechanisms involved in the chemoreception of active compounds, the ecological, physiological and biochemical relevance of the responses and possible application to controlled release technology are discussed.

Amino Acids↗

Two glutathione S-transferase isoenzymes purified from Bulinus truncatus (Gastropoda: Planorbidae).

We purified and characterized two major glutathione S-transferase isoenzymes (GST2 and GST3) from snail Bulinus truncatus (Mollusca, Gastropoda, Planorbidae) tissue. The Km with respect to 1-chloro-2, 4-dinitrobenzene (CDNB) for both isoenzymes was increased as the pH decreased. Km of both isoenzymes with respect to glutathione (GSH) doubled when the pH was increased from 6.0 to 6.5. Acid inactivated GST2 and GST3 and the two enzymes were almost inactive at pH 3.5. However, they retain the full activity for at least 20 h when incubated at pH between 6.0 and 9.0. The optimum temperature was 45 degrees C for GST2 and 50 degrees C for GST3. The half lifetime at 50 degrees C was 70 min and 45 min for GST2 and GST3 isoenzymes, respectively. Addition of 5 mM GSH to the incubation buffer increased the half life of both isoenzymes more than fourfold. The activation energy for catalyzing the conjugation of CDNB was 1.826 and 3.435 kcal/mol for GST2 and GST3, respectively. I50 values for Cibacron blue, bromosulphophthalein, indocyanine green, hematin and ethacrynic acid were 0.76 microM, 47.9 microM, 7.59 microM, 0.03 microM and 0.79 microM for GST2, and 0.479 microM, 79.4 microM, 89.1 microM, 32.4 microM and 1.15 microM for GST3, respectively. Cibacron blue and indocyanine green were non-competitive inhibitors, while hematin was a mixed inhibitor. Bromosulphophthalein was found to be a competitive inhibitor for GST2 and a mixed inhibitor for GST3.

Animals↗

Molluscicidal activity against Bulinus truncatus of Croton campestris.

The bark of Croton campestris provided three furano-clerodane named, respectively, velamone, velamolone acetate and velamolone. Fractions containing these diterpenoids were investigated for their molluscicidal activity against Bulinus truncatus, one of the aquatic snail vectors of schistosomiasis. A dichloromethanic extract of root barks, rich in furano-clerodane and molluscicidal at 20ppm was promising as natural molluscicide. Enriched fraction with velamone gave 80% mortality from 25ppm, while its LD(100) was at 50ppm. The LD(80) was ca. at 45ppm, while LD(100) was at 60ppm for the enriched fraction containing velamolone acetate. The velamolone-enriched fraction was less efficient and gave a mortality of 90% at 90ppm. The pure compounds, velamone and velamolone acetate were active at 100% at 3 and 6ppm, respectively. Velamolone showed a molluscicidal activity at 20ppm.

Animals↗

The influence of self-fertilization and pairing on life-history traits in the freshwater snail Bulinus forskalii (Gastropoda, planorbidae).

This study analyses the basic reproductive biology of the schistosonie-vector snail Bulinus forskalli. This hermaphroditic species can self-fertilize. Variation of the mating system was analysed in two populations from Cameroon in a three-step experiment. (i) The fecundity (number of eggs and of egg capsules) of isolated virgin individuals from both populations was followed over a month, as well as the survival of offspring from these individuals. Estimates of these parameters were intermediate. Our results also indicate that inbreeding depression has a maximum value of about one-half in both populations. No difference was observed between populations for both the overall fecundity of adults and survival of offspring. (ii) Individuals were then paired either within, or between populations over three successive days, and the number of copulations recorded. Copulations were observed in all situations, including between-population pairs. No significant dlifference in the number of copulations per pair was detected. However, this value decreased with time, and reciprocal copulations were significantly less numerous between than within populations. (iii) Individuals that successfully copulated as female in (ii) were isolated, and their fecundity was followed over 3 weeks. The fecundity of individuals that had been maintained isolated over the whole experiment served as a control. No significant difference was on the whole detected. Our results collectively suggest that the individuals from the two populations studied are rather selfers, confirming that B. forskalii is a preferentially selfing species. There is also some pre-zygotic isolation against crosses between populations.

Animals↗

Evaluation of three methods to control Bulinus truncatus, the intermediate host of Schistosoma haematobium in an irrigation scheme, Tessaout-Amont, Morocco.

In Morocco, the implementation of new irrigation schemes resulted in the introduction of schistosomiasis haematobium to new areas. Canal structures (especially syphon boxes) were found to be favourable habitats for Bulinus truncatus and they constitute important transmission sites. Three methods for snail control in these syphon boxes were assessed in the Tessaout-Amont irrigation scheme. Molluscicide (niclosamide) application resulted in a pronounced reduction in the snail population density, but snails started to recolonise the sites a few months later. Regular brushing of the inner sides of the syphon boxes and covering the syphon boxes, so as to exclude light, resulted in significant reductions in snail numbers. However, the latter methods were not as efficient as the molluscicide.

Animals↗

A comparison of Bulinus africanus group species (Planorbidae; Gastropoda) by use of the internal transcribed spacer 1 region combined by morphological and anatomical characters.

The morphological and anatomical characters applied to determine species identity in the Bulinus africanus group species are insufficient to unambiguously discriminate between arbitrary species of given populations. In order to solve this problem, four snail populations from Kenya have been investigated morphologically and anatomically, and the species status compared with the result of molecular methods. We have amplified the entire ITS region and found that the investigated populations showed intra-specific genetic polymorphism, thereby giving the taxa an identity which were indistinct when the region was cut with restriction enzymes. Instead, an amplification of the sub-region ITS 1 revealed an unambiguous identification. because the amplification revealed only one single fragment. We also found that the observed heterogeneity of the entire ITS region could be confined to the sub-region ITS 2. Furthermore, the micro-sculpture of the shell and penis to preputium proportion, which are normally applied as morphological characters, might be considered as inadequate, because of the lack of a significant difference in those characters between the two well established species, namely B. africanus and B. nasutus in the Kisumu area. Instead, these two taxa were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to be one single species with a highly variable morphology. This result was further confirmed by random amplified polymorphic DNA (RAPD) data and the mitochondrial cytochome oxidase subunit I (COI).

Animals↗

Molecular characterization of Bulinus globosus and B. nasutus on Zanzibar, and an investigation of their roles in the epidemiology of Schistosoma haematobium.

The ribosomal internal transcribed spacer (ITS) regions of both Bulinus globosus and B. nasutus from Zanzibar were amplified and restricted with 6 enzymes. Restriction products were separated by polyacrylamide gel electrophoresis and visualized by ethidium bromide staining. Considerable genetic variation was observed at 3 levels: (i) between species, (ii) among populations and (iii) among individuals. In addition, the 18S ribosomal deoxyribonucleic acid gene was amplified from both species and the products were also restricted with several enzymes. No variation was detected. Nucleotide diversity of the ITS was estimated to be approximately 0.019 for both species and nucleotide divergence between species was estimated to be 0.03. Restriction profiles differentiated these snail species and may prove useful for identification purposes. Only B. globosus populations were found to be naturally infected with Schistosoma haematobium. From laboratory infection studies of progeny snails, B. nasutus appeared to be refractory to S. haematobium, whereas certain B. globosus populations were susceptible to laboratory challenge.

Animals↗

Studies of the relationships between Schistosoma and their intermediate hosts. II. The genus Bulinus and Schistosoma haematobium from Sudan, Zaire and Zambia.

The relationship between Schistosoma haematobium from Sudan, Zaire and Zambia and various species and strains of Bulinus, has been investigated. The main emphasis was placed on evaluating the total cercarial production per 100 exposed snails as an index of the compatibility between snail and schistosome. The observations confirmed the results of previous compatibility studies which showed that there were two groups of S. haematobium, truncatus-borne from Sudan and Zaire, and africanus-borne, from Zambia. They also revealed that this division was not absolute, as some overlapping occurred. However, if the cercarial production was taken as an indication of compatibility, instead of the infection rate, there was a more definite division between the two groups. The observation that there is a truncatus-borne strain of S. haematobium present in Zaire extends the area where there is a mixture of truncatus- and africanus-borne strains farther southwards.

Adult↗

Studies of the relationships between Schistosoma and their intermediate hosts. IV. The genus Bulinus and Schistosoma bovis from Morocco.

Schistosoma bovis from Morocco was used in infection experiments with several populations of Bulinus truncatus. The snails from Libya, Malawi, Morocco and Senegal were very compatible with the schistosome since the infection rates were approximately 90%, the mortality was low, and a very high production of cercariae, approximately 1.2 million per 100 exposed snails, was observed. Only a very few B. truncatus (2n = 36) from Rhodesia became infected. B. permembranaceus and B. forskalii were refractory.

Animals↗

The effect of light and darkness on the production of cercariae of Schistosoma haematobium from Bulinus globosus.

The cercariae of Schistosoma haematobium showed a diurnal periodicity of emergence from Bulinus globosus in a twelve hour light/dark cycle. Peak emission occurred at 11.00 hrs with a smaller peak at 20.00 hrs, following the start of the period of darkness. In continuous illumination this second peak was not seen, indicating that only the morning peak is circadian in origin. The evening peak occurs in response to dark treatment and can be produced by periods of darkness ranging from eight seconds to one hour. The longer the period of dark treatment the longer the rise in output is maintained on return to light conditions. Subjection of snails to periods of dark treatment during the normal light period caused a reduction in the evening peak with the largest effect seen following the longest period of darkness. An increased output of cercariae was seen following fifteen minutes exposure to a range of light intensities, the largest increase occurring at 10,000 and 7000 lux and complete darkness. The rapidity of this reaction to variations in light intensity suggests that the cercariae of S. haematobium are showing emergence in response to shadows.

Animals↗

Cercarial shedding patterns of Schistosoma bovis and S. haematobium from single and mixed infections of Bulinus truncatus.

The cercarial shedding of Schistosoma bovis and S. haematobium were studied in single and mixed infections in the snail host Bulinus truncatus. The two species displayed a distinctive diurnal cercarial emergence with an earlier shedding pattern for S. bovis than S. haematobium (the average emergence peaks were respectively at 0800 h and 1200 h). In mixed infections, each species kept its own cercarial shedding rhythm with no marked alterations. The cercarial emergence pattern is proposed as a new method to identify natural mixed infections in the snail intermediate hosts. The interactions between the two parasites are discussed.

Animals↗

The effect of infection with Schistosoma margrebowiei on the growth of Bulinus natalensis.

Three age groups of Bulinus natalensis, immature, mature but not yet egg-laying and mature egg-laying, were infected with miracidia of Schistosoma margrebowiei. The growth of infected, exposed but showing no signs of infection, and uninfected control groups, were examined at weekly intervals for ten weeks post exposure. Snails exposed to infection when immature, even in the group where no patent development ensued, showed a statistically significant reduction in growth and this was evident as early as 2-5 weeks post exposure. When infection occurred at the stage prior to egg-laying a significant reduction in growth was seen but only in the group which developed a patent infection. This was also the case for the egg-laying group although the reduction in growth was only significant in the final three weeks of the experiment. Thus in all three age groups, growth rate was reduced and the infected snails were significantly smaller at the end of ten weeks compared with the controls. The reasons for these effects are discussed.

Animals↗

Bulinus tropicus, a natural intermediate host for Schistosoma margrebowiei in Lochinvar National Park, Zambia.

Snails, provisionally identified as Bulinus tropicus, on the basis of chromosome number, egg protein profile, AcP and HBDH enzymes of the digestive gland, and radular morphology, from Lochinvar National Park, Zambia have been demonstrated to transmit Schistosoma margrebowiei naturally. The identification of the unpaired male schistosomes was confirmed by PGM and AcP analyses. The observations confirm earlier epidemiological predictions, and add another species of mollusc to the two, B. forskalii and B. scalaris, known to be natural intermediate hosts of S. margrebowiei.

Animals↗

Parasites in Bulinus senegalensis (Mollusca: Planorbidae) and their detection.

Isoelectric focusing studies on enzyme variation between populations of the snail Bulinus senegalensis revealed that parasitic infections in the snails contributed additional bands of enzyme activity, particularly in the glucose phosphate isomerase (GPI) and malate dehydrogenase (MDH) systems. The patterns due to the parasite enzymes were, in most cases, clearly distinct from those of the host and different from each other. Parasites encountered included Schistosoma haematobium, S. bovis, Paramphistomum microbothrium, another amphistome probably belonging to the group which infect amphibians, Echinostoma revolutum, another echinostome (probably Echinoparyphium sp.), strigeids, xiphidiocercariae (these were resolved into 3 distinct types by the enzyme data) and ciliate protozoa. The 7 host populations which were examined showed marked differences in both the prevalence and variety of their parasitic infections and these variations were tentatively related to environmental differences in their respective habitats and to the nature of human contact patterns. Seasonal changes in the parasite fauna were also noted and some of the implications of the parasite load on the host population are briefly mentioned.

Animals↗

Spatial and temporal heterogeneity in the population dynamics of Bulinus globosus and Biomphalaria pfeifferi and in the epidemiology of their infection with schistosomes.

Populations of Bulinus globosus and Biomphalaria pfeifferi were studied in a river habitat in Zimbabwe over a period of 12 months. Data were obtained on the prevalences of infections of Schistosoma haematobium (also S. mattheei) and S. mansoni respectively. Population parameters showed the following patterns for both snail species. (1) A patchy distribution correlated with the distributions of aquatic plants. (2) Life-expectancies of only a few weeks. (3) Recruitment rates correlated with water temperature and showing a distinct seasonal peak. (4) Spatial variation in recruitment. (5) A redistribution of snails during the rainy season. Epidemiological parameters showed the following patterns. (1) Large seasonal variations in the prevalence of patent infections. (2) Evidence from size-prevalence curves that suggests a variable force-of-infection from man to snail, correlated with water temperature. (3) Prevalences of infection that were higher in the vicinity of (+/- 60 m from) major water contact sites. Local prevalences of infection for B. globosus sometimes exceeded 50% and may have approached 100% if pre-patent infections are included. Snail numbers may limit transmission at these locations. Attention is drawn to the need to make field observations at an appropriate spatial scale and also to the implications for the effectiveness of focal snail control as a means of reducing transmission.

Animals↗

Effects of Schistosoma haematobium infection on reproductive success and male outcrossing ability in the simultaneous hermaphrodite, Bulinus truncatus (Gastropoda: Planorbidae).

The schistosome intermediate snail host, Bulinus truncatus (Mollusca: Planorbidae), has two reproductive (phally) morphs. Both aphallics and euphallics can self-fertilize, but aphallics cannot donate sperm because they do not develop a functional penis and prostate. This study investigated the interactions between phally and fitness consequences of Schistosoma haematobium infection in B. truncatus. Snails which developed patent infections produced 26% fewer eggs than controls and 35% fewer eggs than exposed snails which did not develop infections. This reduction was due to a lower lifetime production of egg masses and a smaller mean number of eggs/mass in infected snails relative to control or exposed snails. However, there was no evidence of increased mortality in infected snails. Contrary to reports of fecundity compensation in other intermediate host snails, egg production post-exposure during the pre-patent period did not increase relative to that of controls in either infected or exposed snails. Phally did not influence susceptibility to infection or length of the prepatent period. Furthermore, lifetime egg, egg mass and hatchling production, as well as mean eggs/mass and number of hatchlings reaching maturity, did not differ significantly between aphallics and euphallics within control of exposed experimental groups. However, within the infected group euphallics produced 38% fewer eggs, smaller egg masses, and fewer hatchlings reaching maturity than aphallics, supporting the prediction of a cost to the growth and maintenance of a full male tract. This cost was detectable only when snails were under the stress of infection. The proportion of euphallic offspring produced did not differ across experimental groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗