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Two exemplary cases of mixed urogenital and intestinal schistosomiasis in Malawi show hybridizations of Schistosoma mattheei with S. haematobium and S. mansoni, indicating newly emerging genetic diversity. Complex egg excretion patterns in feces expose current diagnostic gaps and alert to future sampling needs for effective surveillance of zoonotic schistosomiasis.

Background Male Genital Schistosomiasis (MGS) remains an often-overlooked chronic sequela of urogenital schistosomiasis in endemic areas of sub-Saharan Africa. As part of a 2-year longitudinal study on Hybridization of UroGenital Schistosomiasis (HUGS) in Malawi, a MGS sub-study was conducted to assess whether hybrid schistosomes were incriminated. Methods During recruitment, demographic, health and socio-economic data were collected through individual questionnaire interviews in Mthawira community from Nsanje District along Shire River and Samama community from Mangochi District along Lake Malawi shoreline. Urine and semen samples were collected and analysed to determine the identity of schistosome infection. Urine filtration and microscopy, direct microscopy of semen and its sediments (after centrifugation) were performed. Thereafter, the sediments were examined by molecular DNA analysis with a novel two-tube real-time PCR assay. The participants were also screened for Human papilloma virus (HPV) and other sexually transmitted infections (STIs). Results Twenty-two men were recruited for the sub-study, 8 in Nsanje District and 14 in Mangochi District, with a median age of 22.0 years. By microscopy, ten (45.7%) participants had Schistosoma ova in their urine, 11 (50.0%) in semen while 16 (72.7%) were positive by real-time PCR. One participant had both S. haematobium and S. mattheei ova in his semen, three showed symptoms, and one had a mixed infection of S. mansoni and possible S. haematobium - S. mattheei hybrid. Twelve men had detectable high-risk HPV serotypes 16, 18 and others while six had Trichomonas vaginalis and other STIs. Conclusion Zoonotic and hybrid schistosomes can cause MGS similar to human schistosomes, which can be co-infected with HPV and STIs, thereby posing a new challenge in diagnosis, management and control measures in resource poor settings. Increased awareness of these infections among local communities and primary healthcare workers and improvement of disease management are needed and advocated.

Our pilot parasitological investigation of cattle, supplemented with molecular DNA characterisation of encountered schistosomes, sheds first light upon bovine schistosomiasis on Unguja Island, Zanzibar. During February 2024, a total of 99 cattle were examined. Of these, 47 were exported animals from the Tanzanian mainland, designated for slaughter at two governmental abattoirs (Kisakasaka and Muwanda), and 52 were free-grazing animals sampled from four grazing locations within the island’s North and West-B regions. Upon visual inspection of 31 cattle carcasses at Kisakasaka for adult worms, the prevalence of bovine schistosomiasis was 51.6%; however, upon faecal miracidia hatching test (MHT) it was 80.6%. At Muwanda, only faecal MHT was used, finding a much lower prevalence of 12.5%. In free-grazing animals, the prevalence of bovine schistosomiasis by MHT was 0.0%. At Muwanda, the animal quarantine paddock was in disrepair, inclusive of a large pond now acting as a watering point. Here, numerous Bulinus forskalii sp. were found. Whilst no snails were observed to shed schistosome cercariae, molecular xenomonitoring did detect a pre-patent infection prevalence of 10.8%, with Schistosoma bovis firmly incriminated. Molecular DNA characterisation of adult schistosomes ( n  = 19) by real-time polymerase chain reaction (PCR) and high-resolution melt profiling, alongside DNA sequencing, also identified S. bovis , although two worms were putative S. bovis-S. mattheei hybrids. Atypical intrauterine eggs of S. bovis were noted upon microscopy of a worm pair. A broader screen of 92 miracidia confirmed S. bovis and three miracidia as S. bovis-S. mattheei hybrids. Contrasting with Pemba Island, Zanzibar, where autochthonous transmission of S. bovis can occur, bovine schistosomiasis on Unguja Island currently appears restricted to imported animals alone. However, the seminal detection of putative S. bovis-mattheei hybrids, alongside the current inadequate quarantine facilities at Muwanda, raises a new concern that such hybrid schistosomes may escape and enter the island’s hinterland. Should this happen, surveillance and control of urogenital schistosomiasis on Unguja would be compromised and further complicated. We therefore strongly recommend immediate repair and improved maintenance of governmental animal quarantine facilities. Future epidemiological surveys of imported cattle are now well justified, not only to better understand the full repertoire of hybrid schistosomes present but also to develop appropriate mitigating interventions. Graphical Abstract

Background Snail-borne trematodes afflict humans, livestock, and wildlife. Recognizing their zoonotic potential and possible hybridization, a One Health approach is essential for effective control. Given the dearth of knowledge on African trematodes, this study aimed to map snail and trematode diversity, focusing on (i) characterizing gastropod snail species and their trematode parasites, (ii) determining infection rates of snail species as intermediate hosts for medically, veterinary, and ecologically significant trematodes, and (iii) comparing their diversity across endemic regions. Methods A cross-sectional study conducted in 2021 in Chiredzi and Wedza districts in Zimbabwe, known for high human schistosomiasis prevalence, involved malacological surveys at 56 sites. Trematode infections in snails were detected through shedding experiments and multiplex rapid diagnostic polymerase chain reactions (RD-PCRs). Morphological and molecular analyses were employed to identify snail and trematode species. Results Among 3209 collected snail specimens, 11 species were identified, including schistosome and fasciolid competent snail species. We report for the first time the invasive exotic snail Tarebia granifera in Zimbabwe, which was highly abundant, mainly in Chiredzi, occurring at 29 out of 35 sites. Shedding experiments on 1303 snails revealed a 2.24% infection rate, with 15 trematode species identified through molecular genotyping. Five species were exclusive to Chiredzi: Bolbophorus sp., Schistosoma mansoni , Schistosoma mattheei, Calicophoron sp., and Uvulifer sp. Eight were exclusive to Wedza, including Trichobilharzia sp., Stephanoprora amurensis , Spirorchid sp., and Echinostoma sp. as well as an unidentified species of the Plagiorchioidea superfamily. One species, Tylodelphys mashonensis , was common to both regions. The RD-PCR screening of 976 non-shedding snails indicated a 35.7% trematode infection rate, including the presence of schistosomes (1.1%) Fasciola nyanzae (0.6%). In Chiredzi, Radix natalensis had the highest trematode infection prevalence (33.3%), while in Wedza, R. natalensis (55.4%) and Bulinus tropicus (53.2%) had the highest infection prevalence. Conclusions Our xenomonitoring approach unveiled 15 trematode species, including nine new records in Zimbabwe. Schistosoma mansoni persists in the study region despite six mass deworming rounds. The high snail and parasite diversity, including the presence of exotic snail species that can impact endemic species and biomedically important trematodes, underscores the need for increased monitoring. Graphical Abstract

Terminal-spined Schistosoma sp. eggs were detected in several groups of baboons living in Kafue National Park in central Zambia. A total of 166 fecal samples was screened; egg prevalence overall ranged between 7% and 10%, while infection intensities were low. Formalin-fixed eggs had an average length of 144.5 µm and a breadth of 48.3 µm, but the schistosome species could not be unambiguously identified by size or morphology. We used molecular methods to definitively identify the parasite species. Parasite DNA was amplified from stools by polymerase chain reaction (PCR). Sequence analysis of fragments of the first internal transcribed spacer (ITS-1), mitochondrial 12S rDNA, NADH dehydrogenase subunit 6 (nad6), and cytochrome C oxidase subunit 1 (cox1) from 3 egg-positive samples revealed the presence of S. mattheei in these samples. This is the first molecular identification of S. mattheei from free-ranging baboons. Schistosoma mattheei is typically a parasite of bovids, but it can also infect humans. Schistosoma mattheei in baboons in Zambia may affect other wildlife species and humans that live in close proximity to baboons.

The susceptibility of Bulinus tropicus , B. globosus , Biomphalaria pfeifferi , Lymnaea natalensis , and Melanoides tuberculata to Calicophoron microbothrium was examined. Bulinus tropicus had the highest prevalence (65.0%), followed by B. pfeifferi (37.5%), B. globosus (6.8%), and M. tuberculata (5.9%). Lymnaea natalensis was refractory to infection. Bulinus tropicus snails infected with C. microbothrium alone or coinfected with either Schistosoma haematobium or S. mattheei 0, 7, 14, and 21 days after exposure to C. microbothrium produced C. microbothrium cercariae only.

We reviewed the early literature and maps of the occurrence of urogenital schistosomiasis (bilharzia) in the Eastern Cape, South Africa from the 1860s until its decline from about 1900 and reappearance in 2002. Although this decline in transmission has received little attention to date, clinical descriptions of the disease over this period indicate that infection was common, probably patchy, although sometimes with severe morbidity. The long period of quiescence between 1900 and 2002 is thought to be as a result of several factors, but primarily because of the impact of the area's cold winters and drought-prone climate on the survival and reproduction of both the snail intermediate host Bulinus africanus and the intramolluscan stages of the parasite. The concept of an outbreak area is invoked to describe the occurrence of intense urogenital schistosomiasis transmission in localised areas for relatively short periods of up to 35 years in this the southernmost part of its range in Africa, a suboptimal environment for transmission.

The study investigated whether the susceptibility of calves to an early Schistosoma mattheei infection may be modified by intake of colostrum from infected cows. Twelve calves born to non-infected mothers were randomly divided into 2 groups of 6. The animals from group 1 were fed colostrum originating from a pool collected from non-infected cows, the calves from group 2 received colostrum from a pool collected from cows infected with S. mattheei. One month after birth all calves were infected by exposure to 1000 cercariae of a local strain of S. mattheei, and perfused 12 weeks later to determine the worm- and tissue egg counts. IgGH+L, IgG1, IgG2 and IgA levels against soluble adult worm antigen preparation of S. bovis (SWAP bovis) were analysed in both colostrum pools and in the serum from the calves collected during the study before and after receiving colostrum, then on days 7, 30, 73 and 122. Faecal egg counts were determined from day 73 onwards. The IgGH+L, IgG1 and IgA levels of the positive colostrum pool were higher than those of the negative pool. Calves of group 2 showed significantly higher levels of IgGH+L and IgG1 until day 73, to reach equal levels at necropsy. Calves of group 2 showed significant reductions of 42, 28 and 42% in total worm counts, female worm counts, and tissue egg counts, respectively, and a reduction of 25% in cumulative faecal egg counts. These findings indicate that there was a significant impact of colostrum on the parasitological and serological course of early S. mattheei infections.

Over a period of about 12 years, 30 abnormal Schistosoma mattheei cercariae were found among a total of approximately 2.8 million examined. Initially seven were recovered from about 1.02 million (0.0007 %), which were examined individually while being counted with the aid of a stereoscopic microscope. Subsequently, on the strength of relatively high percentages of abnormal individuals recovered when counting cercariae that failed to penetrate into oxen, it appeared that the morphologically abnormal cercariae were unable to swim and would mostly sediment out of a suspension while most of the normal cercariae would remain swimming. This surmise is supported by recovery of 23 morphologically abnormal cercariae (0.001 %) from about 1.8 million, by examining the sediment after the cercarial suspension had been left standing undisturbed in glass measuring cylinders. The abnormalities ranged from aberrant tails only (e.g. an underdeveloped tail, or different degrees of schism) or aberrant heads only, to abnormalities of both the heads and tails. A suggested schematic classification of abnormal cercariae is presented. A young, adult hamster was exposed to eight S. mattheei cercariae with complete schism of the shaft of the tail, by pipetting the cercariae onto the shaved abdominal skin of the anaesthetised animal. Two underdeveloped females were subsequently encountered in squash preparations of the liver when the hamster was killed for worm recovery 10 weeks after infection, thus showing that some of the abnormal cercariae were viable. A method is also described for killing and fixing cercariae while retaining some of the shining brilliance of live cercariae, without them becoming shrivelled, granular and semi-opaque, as occurs when cercariae die spontaneously or are killed with heat This is apparently the first report of abnormal cercariae of S. mattheei. In addition, a method of concentrating abnormal cercariae after emergence from a snail, a schematic classification of abnormal cercariae and a method for killing and fixing cercariae while retaining much of the shiny brilliance of live cercariae are also reported for the first time as far as is known.