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.     

Studies on the relationship between Schistosoma and their intermediate hosts. V. The genus Bulinus and Schistosoma bovis from Iringa, Tanzania

The relationship between an isolate of Schistosoma bovis from Iringa, Tanzania, and various species of the host snail genus Bulinus from East Africa was studied using the total cercarial production per 100 exposed snails over a period of 4 weeks following patency as an index of the compatibility. All populations of Bulinus forskalii and B. africanus tested exhibited a high level of susceptibility while the populations of B. truncatus and B. globosus tested were either refractory or of low to moderately low susceptibility. All populations of B. abyssinicus, B. canescens, B. nasutus and B. tropicus tested were refractory. It is suggested that B. africanus is the most important host snail for S. bovis in East Africa, that B. forskalii at least locally may contribute significantly to the transmission and that B. truncatus and B. globosus only play a limited role in the transmission.     

Developing species-specific primers to identify Bulinus truncatus and Bulinus beccari, the intermediate hosts of Schistosoma haematobium in Saudi Arabia

This work aimed to determine the inter- and intra-specific variations in populations of Bulinus truncatus and Bulinus beccari, the intermediate hosts of Schistosoma haematobium in Saudi Arabia, and to develop species-specific primers to identify these snails as a first step in the development of multiplex PCR for simultaneously identifying the snails and diagnosing its infections in a single step. Two populations of B. truncatus were collected from Asser and Bisha (A and B), and two B. beccari populations were collected from Mahial Asser and Merba (C and D). The snails' genomic DNA was extracted and amplified using 5 different primers. The primers displayed variable intra- and inter-specific differences across the populations. The largest RAPD-PCR fragments were cloned into a vector as a preparatory step for sequencing. Similarity searches for the sequenced cloned inserts revealed no similar sequences in the GenBank database or its associated databases. Specific primers used to target the B. truncatus and B. beccari genomes were designed using the Gene Runner program and based on the DNA sequences obtained from RAPD fragment sequence analyses. Using these primers for specific PCRs resulted in expected single-band PCR products of 536 bp for B. beccari and 478 bp for B. truncatus. These results will be helpful for simultaneously identifying B. truncatus and B. beccari snails and diagnosing S. haematobium infections within the snails using single step multiplex PCR.     

Studies of the relationship between Schistosoma and their intermediate hosts. III. The genus Biomphalaria and Schistosoma mansoni from Egypt, Kenya, Sudan, Uganda, West Indies (St. Lucia) and Zaire (two different strains: Katanga and Kinshasa)

The compatibility between strains of Schistosoma mansoni from Egypt, Kenya, Sudan, Uganda, the West Indies, and Zaire (two strains which came from Katanga and from Kinshasa), and various species and strains of Biomphalaria, i.e. Biomphalaria pfeifferi, B. alexandrina, B. glabrata and B. camerunensis was investigated. Data as mortality, rate of infection of the surviving snails, duration of infection, cercarial production per day per positive snail, etc., were observed. The main emphasis was placed on determining the total cercarial production per 100 exposed snails for each snail population. It was possible to infect all the tested populations of B pfeifferi with the various strains of S. mansoni, but the observation as e.g. TCP/100 exposed snails varied greatly according to the population of snail and the strain of S. mansoni. The results for the remaining species of Biomphalaria varied greatly, depending on the combination, e.g. B. alexandrina was only susceptible to the local S. mansoni from Egypt. The highest TCP/100 exposed snails was more than 1 million for the strains of S. mansoni from Egypt, Kenya and the West Indies in B. alexandrina, B. pfeifferi and B. glabrata, respectively. The next group, with a TCP/100 exposed snails on 7--800 000 consists of S. mansoni from Sudan, Uganda and Zaire (Katanga) all in B. pfeifferi. The last tested strain of S. mansoni, Zaire (Kinshasa) yielded a cercarial production on 500 000 per 100 exposed snails in B. pfeifferi and B. camerunensis. The shortest prepatent period, 19 days, was observed for S. mansoni from Kinshasa, Zaire, in B. camerunensis, and the longest prepatent period, 25 days, was found for strains from Egypt and from the West Indies in B. alexandrina and B. glabrata, respectively. In general, a very long duration of infection, lasting up to 200 days, was observed.     

Discussion of the relationships between Schistosoma and their intermediate hosts, assessment of the degree of host-parasite compatibility and evaluation of schistosome taxonomy.

Studies of the spectrum of the intermediate hosts of schistosomes and the relationships between parasites and intermediate host snails have been carried out with each researcher using different methods. A comparison between compatibility studies has therefore been impossible. The following three basic problems in these kind of studies are discussed in this paper: (1) the standardization of experimental materials such as schistosomes, intermediate and final hosts, (2) the methods involved in the experiments, and (3) the sort of data which must be collected. Standardized methods and materials are described in this paper. The relationships between the genus Schistosoma and the intermediate host snails belonging to the two genera Biomphalaria and Bulinus, and the mechanisms behind the process resulting in various degrees of compatibility between the two involved organisms are reviewed and discussed. Why certain combinations of schistosomes and snails result in the production of cercariae and others do not is still unknown, but there is now some indication of a camouflage behaviour during the intramolluscan stages, e.g., the larvae in the snails cover themselves with snail material and are not recognized as foreign objects. The same mechanism has already been found in the schistosomules in the final host. Much of the confusion in the discussion of the relationships between schistosomes and intermediate hosts is the result of a lack of an objective way to describe and assess the compatibility between the two. Based on a series of experiments with S. haematobium, S. intercalatum, S. bovis, S. mansoni, and several species of possible intermediate hosts, the following index for estimating the degree of compatibility is proposed: the total cercarial production from 100 exposed snails (TCP/100 exposed snails). Seven classes have been suggested: the first one (Class 0) with a TCP/100 exposed snails of zero, is called 'refractory'; the next one (Class I) called 'not very compatible' has a TCP /100 exposed snails between 1 and 10,000; the last group (Class IV) called 'extremely compatible' produced more than 500,001 per 100 exposed snails during the entire lifespan. The ability to use a spectrum of intermediate hosts and the compatibility in the taxonomy of schistosomes is discussed. The results for the different species of schistosomes indicated that each species consists of strains with 'hybrid populations' in between.     

Spatial and temporal population genetic survey of Bulinus globosus from Zanzibar: an intermediate host of Schistosoma haematobium

In Zanzibar, the freshwater hermaphroditic snail Bulinus globosus is the intermediate host of the human parasite Schistosoma haematobium . To shed light on the patterns of genetic variation within Bu. globosus , variation at six polymorphic microsatellite loci was assessed to quantify spatial genetic structure within eight populations and temporal changes in a further four populations of the snails. Limited genetic variation was observed within populations, possibly reflecting both partial-selfing and fluctuations in population size. Although a statistically significant heterozygote deficiency was observed, Bu. globosus appears to be a preferential out-crosser, which was consistent with the absence of genotypic linkage disequilibria. Genetic variation across populations was substantial, illustrating significant isolation-by-distance on a regional scale and that gene flow was restricted to nearby populations. The temporal survey showed that levels of genetic variation and inbreeding changed over a 1-year period, consistent with field observations of seasonal change. The results strongly support a snail demographic model of population expansion and contraction throughout the year with habitats re-colonized by aestivating or surviving snails from local refugia. This model might help explain co-evolution of snails and schistosomes in Zanzibar as local populations of Bu. globosus have varying degrees of susceptibility to S. haematobium .     

Field observations on the population dynamics of Bulinus globosus, the intermediate host of Schistosoma haematobium in the Ifakara area, Tanzania

The population dynamics of Bulinus globosus were studied in a stream and an adjacent pool near Ifakara, Tanzania. Experiments with caged snails were used to determine fecundity, hatching rates, growth and mortality. Growth and mortality rates were also estimated for the natural population from successive size frequency distributions. Oviposition began in snails as small as 6.6 mm and there was a positive correlation between the size of the snails at oviposition and the number of embryos per egg mass. Fecundity was season-dependent. In the pool, snail growth was constant throughout the year, while in the stream, growth was markedly reduced during the rainy season. Mortality rates correlated with fluctuations in snail population density and were highest during the rainy season and among young snails. The high intrinsic rate of natural increase observed for B. globosus enables it to exist in habitats which alternate frequently between favorable and adverse conditions.     

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.     

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.     

Studies of the relationships between Schistosoma nasale and S. spindale and their snail host Indoplanorbis exustus

Infectivity and cercarial production of Indoplanorbis exustus related to variation of miracidial dose (1, 4, 10 or 20) with Schistosoma nasale and S. spindale from Sri Lanka were studied. The intermediate host-parasite relationships of the two schistosome species showed marked differences under the conditions of observation recorded in this study. Prepatent death rates (PDR) were on average higher for S. spindale (30%) than for S. nasale (10%). The size of the miracidial dose to which snails had been exposed had no effect on PDR. The infection rates (IR) were on average higher for S. nasale (41%) compared with S. spindale (27%). Highest IR occurred after exposure to 4 miracidia in S. nasale infections (79%) and after exposure to 10 miracidia in S. spindale infections (60%). The highest daily average cercarial production per snail was recorded for S. nasale at a level of 4 miracidia (1311), and for S. spindale at a level of 10 miracidia (1615). At low level (1 or 4 miracidia) of exposure, I. exustus showed a better compatibility with S. nasale than with S. spindale. An opposite tendency was observed at higher levels (10 or 20 miracidia) of exposure. Unsuccessful infections of Lymnaea luteola with either S. nasale or S. spindale indicate that this species is not involved in transmission.     

Schistosoma mansoni, S. japonicum, S. haematobium: glycogen content and glucose uptake in parasites from fasted and control hosts

The glycogen content of male and female Schistosoma mansoni has been measured in flukes from normally fed hosts and those from fasted hosts. In infections from both the mouse and the hamster, a significant reduction in schistosomal glycogen of males is seen hours after food is withdrawn from the host. Reductions in protein content of the schistosomes were only observed in hamster infections fasted at least 72 hr. The livers of infected mice not only decrease in size during fasting, but there is a concomitant reduction in glycogen per unit wet weight. Comparisons of glycogen:protein ratios of mansonian males, females, and host livers indicate that the fasting-induced loss of liver glycogen is also observed in the male schistosome, but not the female. Studies of both S. mansoni and S. haematobium pairs from fed hosts suggest that the ratio of glycogen:protein contents in the male schistosome correlates with the glycogen:protein ratio of the female partner. Measurements of glucose uptake in vitro suggest that greater uptake rates may be observed in flukes perfused from fasted hosts. In S. japonicum from infected mice, a reduction in male glycogen was also detected as early as after a 6-hr fasting period, but changes in the females were not significant. Unmated male S. japonicum also exhibit a reduction in glycogen levels after fasting, but the quantity of worm glycogen present in these males remains higher than comparable mated males. In mice entrained to a regulated pattern of available food, fluctuations in glycogen content of the male schistosomes were observed, but in the female partners fluctuations were of a smaller magnitude.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Some peculiarities of the relationships between parasitic copepods and their invertebrate hosts

According to the rule of academican E. N. Pavlovskiy, any organism of host is an environment of inhabit for a parasite (Pavlovskiy, 1934). It was analysed, which "ecological niche" or microbiotop (= microhabitat) is occupied by this or that species of symbiotic (parasitic) copepods in organisms of different groups invertebrate-hosts. The assumption lying in a basis of the given analysis means that each group of hosts may give to cohabitants only certain variants of microbiotopes independently on the general morphological structure and life mode of hosts. Five types of microbiotops offered by various groups of hosts for symbiotic copepods are designated (Ta[symbol: see text] 2). 1. The body surface of benthic invertebrates as a microbiotope is characterized by conditions being little different (concerning any kind of physical and chemical influences on copepods) from those in external environment on any other substrate. Apparently a trophical dependence plays a determining role in this case. There are certain directions in a development of adaptations, which are characteristic in some extent for all water ectoparasitic crustaceans and have one functional task--to help to an ectoparasite to keep itself on a surface of host body. In the first, the maxillules and maxillipeds significantly are developed, they get a form of large claws, with which the dopepods are strongly attached on a surface of host body and have an opportunity to move on it without a danger to be washed off. In the second, the form of the body undergoes a dorso-ventral expression and expansion of prosome, forms a cephalic shield allowing to the symbiont to press itself tightly to the host body surface and to avoid the loss of host (tab. 2). In occasions, some ectoparasites stimulate the formation of galls in skin tissues of the host, that also provides the parasite with constant conditions, without any threat to lose the host. However, this phenomenon has not a wide distribution and is observed in some groups of crustacean and echinoderm hosts. 2. The narrow tubular cavities in the organism of host either they are a part of external environment (as in channel system of spongia) or a part of internal environment of organism (as channels of blood system or thin parts of a digestive system) have always rigidly limited sizes and form. Characteristics of all parasites occupying this microbiotopes are the strong transformations. They are expressed by the reduction of legs or any other appendages (frequently in a significant degree), loss of segmentation to some extent and in eruciform (or vermiform) form of a body (tab. 2). This microbiotope is occupied by an ectoparasite in one case only (Spongicola uncifer from channel system of spongia) and by endoparasites in all other cases. 3. Large cavities connected with external environment. The formations of various genesis, such as mantle cavity of molluscs, gill cavity and marsupium of crustaceans, bursal cavity of ophiuroids and branchial cavity of ascidians, concern this type of microbiotopes. All of them are characterized by the relative difference from the external environment and rather large volume (in comparison to sizes of copepods), that provides the parasites with a sufficient protection from factors of the external environment and constant source of food such as elements of host body or food's particles brought by the water flow. Morphological changes in inhabitants of the microbiotope have two directions. They practically are absent in the overwhelming majority copepods, living in the mantle of cavity of the lamellibranches. On the other hand, the inhabitants of gill cavity and marsupium of crustaceans, bursal cavities of ophiuroids and branchial cavity of ascidians are characterized by the presence of strong transformations. Usually there are expressed in a loss of segmentation to some extent, reduction of appendages and swelling of body, as in species of the genus Sphaeronella (tab. 2). Changes are also observed in the life cycle: the tendency to reduce stages of development (development of nauplii stage, which takes place under the ovarial cover). In this case the copepodid stages hatch from the ova. 4. The internal cavity of organism of host. This type of microbiotopes in different groups of the hosts is represented in a various degree. We recognise it in a coelome of polychaetes, lacunar system of molluscs, mixocoel of crustaceans, coelome of echinoderms and cavity of body in ascidians. Two basic evolutionary directions are observed in copepods occupying this microbiotope. In the first case, the parasite is not exposed to transformations and keeps the initial plan of structure as in ancestral free-living forms. In the second case the parasites are exposed to strong transformations, they either live directly in cavity's liquid, or are surrounded by a cyst (as in Cucumaricolidae). 5. Microbiotope of the last type is most specific. The simultaneous existence in two environments--external environment (environment of the second order) and internal environment (environment of the first order) leads to the complete loss of ancestral type in a structure and level of organisation. At the same time both morphological and functional division of the parasite body into two parts produces a new formation--the ectosome and endosome. In this case we deals with the phenomenon of mesoparasitism.     

Schistosomiasis at Loum, Cameroun; Schistosoma haematobium, S. intercalatum and their natural hybrid.

A survey of 500 schoolchildren in Loum in 1968 revealed an overall infection rate of 54.2% with Schistosoma intercalatum and this was the only species of schistosome encountered. In 1972 a number of children were found to be passing schistosome eggs in their urine and these eggs ranged in shape and size from the forms characteristic for S. haematobium to those of S. intercalatum. Preliminary laboratory studies demonstrated that hybridisation between the two species was occurring. Subsequent field surveys showed that the snail hosts for the two parasites (B. rohlfsi for S. haematobium and B. forskali for S. intercalatum) were both present in the river Mbette and its tributaries in Loum and the distribution of the two snail species coincided closely with the distribution of the schistosomes in the human population. Detailed study of a small group of children passing hybrid eggs in their urine revealed that few of them were passing eggs in their faeces and that those eggs which were found in faeces were not viable. Analysis of schistosome egg-shape by plotting cumulative size-frequency data on probability paper demonstrated that the graph obtained from a natural hybrid series was different from that given by a known mixture of the two separate species. The hybrid series included a number of exceptionally large eggs resembling those of S. bovis but isolation of these eggs and subsequent laboratory passage of the parasites showed that they were part of the series and were not evidence of the presence of a third species. Hybridisation experiments in the laboratory showed that the cross S. haematobium male X S. intercalatum femal is fully viable but that the reverse mating is not successful, thus accounting for the failure of the faecal eggs recovered from children with hybrid infections. Histological results from laboratory passaged hybrids suggest that the Ziehl-positive staining reaction of the egg-shells of S. intercalatum may be a recessive character. The observations reported here indicate that S. haematobium has only recently become established in Loum and that it is, through introgressive hybridisation, replacing the indigenous S. intercalatum. A suggested explanation for the change in the parasite fauna is offered and this depends upon ecological changes resulting from forest clearance and agricultural development providing improved conditions for the spread of B. rohlfsi, the snail host for S. haematobium. It is suggested that, in contrast to recent reports on the spread of S. intercalatum, this species is in fact retreating and being replaced by S. haematobium in areas where forest clearance is taking place. In conclusion it is suggested that introgressive hybridisation of this kind may have been responsible for the evolution of certain characteristic local strains of African schistosomes.     

Hybridisation between the digeneans Schistosoma haematobium and S. mattheei : viability of hybrids and their development in sheep

The F1 and F2 hybrids of Schistosoma haematobium male × S. mattheei female were studied with regard to infectivity to intermediate and definitive hosts, isoenzymes (phosphoglucomutase) of individual male worms, randomly amplified polymorphic DNAs of individual adult worms and scanning electron micrographs of the tubercles of male worms. The infection rate of the F1 hybrid miracidia in Bulinus globosus (41.7%) was greater than that achieved in B. wrighti (16.3%); the infection rate of the F2 in B. wrighti was 15.4%. In the definitive hosts: in sheep only male F1 hybrids (i.e. no females and no F2 worms)were recovered; but in hamsters both paired F1 worms and unpaired F1 males were recovered, as were one pair of worms and unpaired males of the F2 generation. The S. mattheei and S. haematobium male worms showed very distinctive PGM patterns, and the F1 hybrids showed additive patterns and a polymorphism with two distinct types of band patterns which are the result of polymorphism in the S. haematobium. The RAPD profiles of the F1 hybrids were also composite of the two parental species. Scanning electron micrographs of the tubercles of male S. haematobium showed them to be heavily spined, whereas those of S. mattheei males were devoid of spines. The F1 hybrids did show variation ranging from non-spined, some with partial spination, to those with heavily spined tubercles. Male worms of the F2 generation possessed tubercles either with or without spines. The potential significance of hybridisation in areas of sympatry between S. haematobium and S. mattheei is discussed.