Diverging patterns of introgression from Schistosoma bovis across S. haematobium African lineages

Hybridization is a fascinating evolutionary phenomenon that raises the question of how species maintain their integrity. Inter-species hybridization occurs between certain Schistosoma species that can cause important public health and veterinary issues. In particular hybrids between Schistosoma haematobium and S. bovis associated with humans and animals respectively are frequently identified in Africa. Recent genomic evidence indicates that some S. haematobium populations show signatures of genomic introgression from S. bovis. Here, we conducted a genomic comparative study and investigated the genomic relationships between S. haematobium, S. bovis and their hybrids using 19 isolates originating from a wide geographical range over Africa, including samples initially classified as S. haematobium (n = 11), S. bovis (n = 6) and S. haematobium x S. bovis hybrids (n = 2). Based on a whole genomic sequencing approach, we developed 56,181 SNPs that allowed a clear differentiation of S. bovis isolates from a genomic cluster including all S. haematobium isolates and a natural S. haematobium-bovis hybrid. All the isolates from the S. haematobium cluster except the isolate from Madagascar harbored signatures of genomic introgression from S. bovis. Isolates from Corsica, Mali and Egypt harbored the S. bovis-like Invadolysin gene, an introgressed tract that has been previously detected in some introgressed S. haematobium populations from Niger. Together our results highlight the fact that introgression from S. bovis is widespread across S. haematobium and that the observed introgression is unidirectional.     

Studies on experimental mixed infections of Schistosoma mansoni and S. haematobium in hamsters

Golden hamsters were superinfected simultaneously with 100 Schistosoma haematobium cercariae, 1 and 3 weeks after initial infection with 100 S. mansoni cercariae. Results indicate that there was a higher degree of resistance to superinfection with S. haematobium at 1 week following initial infection with S. mansoni than that produced in the other two superinfections. This resistance was evidenced by a reduction in the number and size of worms of both species, decrease in S. haematobium egg extrusion per female and by a striking deviation in the egg distribution pattern of both species. Such an early host resistance was not recorded in previous works. Cross-mating was observed but no hybridization took place and the eggs produced were hatchable and typical of their species.     

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.     

Epidemiology of mixed Schistosoma mansoni and Schistosoma haematobium infections in northern Senegal

Due to the large overlap of Schistosoma mansoni- and Schistosoma haematobium-endemic regions in Africa, many people are at risk of co-infection, with potential adverse effects on schistosomiasis morbidity and control. Nonetheless, studies on the distribution and determinants of mixed Schistosoma infections have to date been rare. We conducted a cross-sectional survey in two communities in northern Senegal (n=857) to obtain further insight into the epidemiology of mixed infections and ectopic egg elimination. Overall prevalences of S. mansoni and S. haematobium infection were 61% and 50%, respectively, in these communities. Among infected subjects, 53% had mixed infections and 8% demonstrated ectopic egg elimination. Risk factors for mixed infection - i.e. gender, community of residence and age - were not different from what is generally seen in Schistosoma-endemic areas. Similar to overall S. mansoni and S. haematobium infections, age-related patterns of mixed infections showed the characteristic convex-shaped curve for schistosomiasis, with a rapid increase in children, a peak in adolescents and a decline in adults. Looking at the data in more detail however, the decline in overall S. haematobium infection prevalences and intensities appeared to be steeper than for S. mansoni, resulting in a decrease in mixed infections and a relative increase in single S. mansoni infections with age. Moreover, individuals with mixed infections had higher infection intensities of both S. mansoni and S. haematobium than those with single infections, especially those with ectopic egg elimination (P<0.05). High infection intensities in mixed infections, as well as age-related differences in infection patterns between S. mansoni and S. haematobium, may influence disease epidemiology and control considerably, and merit further studies into the underlying mechanisms of Schistosoma infections in co-endemic areas.     

Schistosoma bovis: patterns of cercarial emergence from snails of the genera Bulinus and Planorbarius

A comparative experimental study of the rhythmic shedding of two geographic strains of Schistosoma bovis cercariae by Bulinus truncatus and Planorbarius metidjensis did not show any differences in emergence patterns. In our opinion, the results support the hypothesis that cercarial emission rhythms are determined primarily by the parasite in these snail/parasite associations.     

Identification of Schistosoma haematobium, S. bovis and S. curassoni by multivariate analysis of cercarial papillae indices

The disposition of cercarial papillae of 68 pre-identified Schistosoma species was established. All the cercariae originated from Africa and Madagascar and were either obtained from natural or experimental infections, and belonged to three species Schistosoma haematobium, S. bovis and S. curassoni. Discriminant analysis was based on nine characters: average values, skewness and kurtosis of three cercarial indices (AD, AL and U) for each sample or isolate. AD, AL correspond respectively to the relative distance between dorsal and lateral papillae. U corresponds to the total number of tail stem papillae. With the exception of two cases of the 68 (one of them corresponding to cercariae shed by a non-African experimentally infected snail), the method enabled discrimination of S. haematobium, S. bovis and S. curassoni.     

The prepatent period and cercarial production of Schistosoma haematobium in Bulinus truncatus (Egyptian field strains) at different constant temperatures

The developmental time of Schistosoma haematobium in Bulinus truncatus snails (field strains) was determined in the laboratory at different constant temperatures between 18 and 32 degrees C. The basic relationship between the length of the minimum prepatent period (y, in days) and the temperature (x, in degree C) is given by the hyperbolic formula y = 295/(x-15.3), 15.3 being the theoretical "developmental null point" and 295 the constant time-temperature product. The shortest prepatency was 17-19 days at 30, 31 and 32 degrees C; at 18 degrees C, cercarial development required at least 106-113 days. The maturation time frequently exceeded the possible minimum by several weeks. No schistosome matured in our experiments at 17 degrees or 33 degrees C. The cercarial release per snail at weekly exposures showed a maximum at 25 degrees C with a geometric mean of 109 cercariae (95% confidence limits 79-149), decreasing to 8 (2-30) at 18 degrees C and 62 (38-100) at 32 degrees C. The absolute maximum of cercariae shed by one snail during 5 h "stimulation" was 2,150 in a 25 degrees C batch, 48 at 18 degrees C and 529 at 32 degrees C. The epidemiological application, the prognosis of the transmission period and the estimation of the transmission potential in relation to climatic conditions are discussed.     

Bulinus tropicus from Central Kenya acting as a host for Schistosoma bovis

One hundred and twelve snails were collected from two habitats on the Mau Escarpment, Kenya and were provisionally identified as Bulinus tropicus from the characteristics of their shell and soft parts, chromosome number (n = 18), electrophoresis of egg protein on cellulose acetate strip and isoelectric focusing of AcP, GPI, HBDH, MDH and PGM digestive gland enzymes. Of the 55 specimens examined alive in London, 10 were infected with amphistome and schistosome larvae, 9 with amphistome larvae and the remainder were uninfected. The GPI and MDH separations of known infected snails showed two distinct areas of activity: host and parasite. Individual hamsters were exposed to schistosome cercariae emanating from each snail with a double infection (apart from one which died prematurely) and examination of the resulting adult worms showed that all were monomorphic for AcP with a band of enzyme activity at pH 6.45, characteristic of Schistosoma bovis. Examination of eggs found in two infections proved to be S. bovis in shape and size. Exposure of laboratory-bred snails of B. tropicus from the Mau Escarpment and other populations of B. tropicus proved negative. Thus, it is suggested that the presence of the amphistome infection may have a suppressive effect on the immune system of the snail, thereby allowing S. bovis to develop.     

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.     

Schistosoma haematobium in Bulinus guernei: Electron microscopy of hemocyte-sporocyst interactions

Electron microscopy has revealed that in Bulinus guernei (Gambian strain) snails infected with Schistosoma haematobium (Egyptian strain) daughter sporocysts and cercariae, two kinds of hemocytes called granulocytes and hyalinocytes are found associated with the sporocysts. Granulocytes are small, numerous, plumbophilic, and amoeboid. They contain lysosome-like granules. Hyalinocytes are large, sparse, less plumbophilic than granulocytes, and have intracellular microfilaments (about 9 nm wide), and few or no pseudopods. They are devoid of lysosome-like granules. Granulocytes and hyalinocytes infiltrate near sporocysts, but only granulocytes interact with sporocyst microvilli by contact. Granulocytes induce a restricted multilamellated encapsulation reaction. Extracellular microfilaments (about 12.5 nm wide), with a regular transverse structure pattern of about 50-nm periodicity, frequently are found along the outer surface of granulocytes located adjacent to sporocysts. Intracellular filamentous structures and a prominent glycocalyx also are features of the seemingly more active granulocytes contiguous with sporocysts. Cell adhesions may occur between surfaces of (1) granulocytes and sporocysts, (2) interdigitating pseudopodial processes of capsular granulocytes, and (3) granulocytes and hyalinocytes.     

Transmission and diversity of Schistosoma haematobium and S. bovis and their freshwater intermediate snail hosts Bulinus globosus and B. nasutus in the Zanzibar Archipelago,United Republic of Tanzania

BackgroundThe Zanzibar Archipelago (Pemba and Unguja islands) is targeted for the elimination of human urogenital schistosomiasis caused by infection with Schistosoma haematobium where the intermediate snail host is Bulinus globosus. Following multiple studies, it has remained unclear if B. nasutus (a snail species that occupies geographically distinct regions on the Archipelago) is involved in S. haematobium transmission on Zanzibar. Additionally, S. haematobium was thought to be the only Schistosoma species present on the Zanzibar Archipelago until the sympatric transmission of S. bovis, a parasite of ruminants, was recently identified. Here we re-assess the epidemiology of schistosomiasis on Pemba and Unguja together with the role and genetic diversity of the Bulinus spp. involved in transmission.Methodology/Principal findingsMalacological and parasitological surveys were conducted between 2016 and 2019. In total, 11,116 Bulinus spp. snails were collected from 65 of 112 freshwater bodies surveyed. Bulinus species identification were determined using mitochondrial cox1 sequences for a representative subset of collected Bulinus (n = 504) and together with archived museum specimens (n = 6), 433 B. globosus and 77 B. nasutus were identified. Phylogenetic analysis of cox1 haplotypes revealed three distinct populations of B. globosus, two with an overlapping distribution on Pemba and one on Unguja. For B. nasutus, only a single clade with matching haplotypes was observed across the islands and included reference sequences from Kenya. Schistosoma haematobium cercariae (n = 158) were identified from 12 infected B. globosus and one B. nasutus collected between 2016 and 2019 in Pemba, and cercariae originating from 69 Bulinus spp. archived in museum collections. Schistosoma bovis cercariae (n = 21) were identified from seven additional B. globosus collected between 2016 and 2019 in Pemba. By analysing a partial mitochondrial cox1 region and the nuclear ITS (1–5.8S-2) rDNA region of Schistosoma cercariae, we identified 18 S. haematobium and three S. bovis haplotypes representing populations associated with mainland Africa and the Indian Ocean Islands (Zanzibar, Madagascar, Mauritius and Mafia).Conclusions/SignificanceThe individual B. nasutus on Pemba infected with S. haematobium demonstrates that B. nasutus could also play a role in the local transmission of S. haematobium. We provide preliminary evidence that intraspecific variability of S. haematobium on Pemba may increase the transmission potential of S. haematobium locally due to the expanded intermediate host range, and that the presence of S. bovis complicates the environmental surveillance of schistosome infections.     

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.     

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.     

Analysis and comparison of cercarial emergence rhythms of Schistosoma haematobium, S. intercalatum, S. bovis, and their hybrid progeny

In identical experimental conditions, the three schistosomes of the terminal spined egg group (S. haematobium, S. intercalatum and S. bovis) showed significant differences in their cercarial shedding patterns. The cercariae of the F1 hybrids, obtained by experimental crosses between these three species, showed the same circadian emergence rhythm (with one peak) as the cercariae of the parental species. However, the mean shedding time of these hybrid parasites (12:14 +/- 1 h 34 min for S. haematobium x S. intercalatum; 09:58 +/- 1 h 24 min for S. haematobium x S. bovis; 08:57 +/- 1 h 19 min for S. bovis x S. intercalatum) was always in advance to the one of their parental species (13:51 +/- 2 h 04 min for S. haematobium; 13:59 +/- 1 h 55 min for S. intercalatum; 10:09 +/- 2 h 04 min for S. bovis). These results are compared with those obtained from crosses between schistosomes with lateral spined eggs, and from crosses between intraspecific chronobiological variants of S. mansoni. They corroborate the genetic determinism of the cercarial emergence of schistosomes. Moreover, such significant differences between the hybrids and their parents in the times of cercarial emission may be of value in the epidemiological research and characterization of naturally occurring hybrids.     

Mating interactions between Schistosoma haematobium and S. mansoni

Schistosoma haematobium and S. mansoni are two medically important schistosomes, commonly occurring sympatrically in Africa and so potentially able to infect the same human host. Experiments were designed to study the mating behaviour of these two species in mixed infections in hamsters. Analysis of the data obtained showed that both heterospecific and homospecific pairs readily form. No significant difference was seen between the two species in their ability in forming pairs, however, S. mansoni showed a greater homospecific mate preference. Analysis of the data using the Mantel-Haenszel test suggests that mating competition does occur between S. haematobium and S. mansoni, the former being the more dominant species. Both species appeared to be able to change mate, with S. haematobium showing a greater ability in taking S. mansoni females away from S. mansoni males when introduced into a pre-established S. mansoni infection highlighting the competitiveness of S. haematobium. The significance of the results is discussed in relation to the epidemiological consequences occurring in Senegal, and other areas where both species are sympatric.