Highlighting male genital schistosomiasis in Malawi.

Highlighting recent literature, we review the epidemiological and clinical importance of male genital schistosomiasis (MGS) in Malawi. We then discuss why individual disease management is an unmet public health challenge and outline how future interventions should be better set within routine services of HIV and men's sexual and reproductive health clinics.

A geospatial analysis of local intermediate snail host distributions provides insight into schistosomiasis risk within under-sampled areas of southern Lake Malawi.

BACKGROUND: Along the southern shoreline of Lake Malawi, the incidence of schistosomiasis is increasing with snails of the genera Bulinus and Biomphalaria transmitting urogenital and intestinal schistosomiasis, respectively. Since the underlying distribution of snails is partially known, often being focal, developing pragmatic spatial models that interpolate snail information across under-sampled regions is required to understand and assess current and future risk of schistosomiasis. METHODS: A secondary geospatial analysis of recently collected malacological and environmental survey data was undertaken. Using a Bayesian Poisson latent Gaussian process model, abundance data were fitted for Bulinus and Biomphalaria. Interpolating the abundance of snails along the shoreline (given their relative distance along the shoreline) was achieved by smoothing, using extracted environmental rainfall, land surface temperature (LST), evapotranspiration, normalised difference vegetation index (NDVI) and soil type covariate data for all predicted locations. Our adopted model used a combination of two-dimensional (2D) and one dimensional (1D) mapping. RESULTS: A significant association between normalised difference vegetation index (NDVI) and abundance of Bulinus spp. was detected (log risk ratio - 0.83, 95% CrI - 1.57, - 0.09). A qualitatively similar association was found between NDVI and Biomphalaria sp. but was not statistically significant (log risk ratio - 1.42, 95% CrI - 3.09, 0.10). Analyses of all other environmental data were considered non-significant. CONCLUSIONS: The spatial range in which interpolation of snail distributions is possible appears < 10km owing to fine-scale biotic and abiotic heterogeneities. The forthcoming challenge is to refine geospatial sampling frameworks with future opportunities to map schistosomiasis within actual or predicted snail distributions. In so doing, this would better reveal local environmental transmission possibilities.

Development, validation, and pilot application of a high throughput molecular xenomonitoring assay to detect Schistosoma mansoni and other trematode species within Biomphalaria freshwater snail hosts.

Schistosomiasis is a neglected tropical disease (NTD) caused by infection with parasitic trematodes of the genus Schistosoma that can lead to debilitating morbidity and mortality. The World Health Organization recommend molecular xenomonitoring of Biomphalaria spp. freshwater snail intermediate hosts of Schistosoma mansoni to identify highly focal intestinal schistosomiasis transmission sites and monitor disease transmission, particularly in low-endemicity areas. A standardised protocol to do this, however, is needed. Here, two previously published primer sets were selected to develop and validate a multiplex molecular xenomonitoring end-point PCR assay capable of detecting S. mansoni infections within individual Biomphalaria spp. missed by cercarial shedding. The assay proved highly sensitive and highly specific in detecting and amplifying S. mansoni DNA and also proved highly sensitive in detecting and amplifying non-S. mansoni trematode DNA. The optimised assay was then used to screen Biomphalaria spp. collected from a S. mansoni-endemic area for infection and successfully detected S. mansoni infections missed by cercarial shedding as well as infections with non-S. mansoni trematodes. The continued development and use of molecular xenomonitoring assays such as this will aid in improving disease control efforts, significantly reducing disease-related morbidities experienced by those in schistosomiasis-endemic areas.

Modelling the age-prevalence relationship in schistosomiasis: A secondary data analysis of school-aged-children in Mangochi District, Lake Malawi.

Schistosomiasis is an aquatic snail borne parasitic disease, with intestinal schistosomiasis (IS) and urogenital schistosomiasis (UGS) caused by Schistosoma mansoni and S. haematobium infections, respectively. School-aged-children (SAC) are a known vulnerable group and can also suffer from co-infections. Along the shoreline of Lake Malawi a newly emerging outbreak of IS is occurring with increasing UGS co-infection rates. Age-prevalence (co)infection profiles are not fully understood. To shed light on these (co)infection trends by Schistosoma species and by age of child, we conducted a secondary data analysis of primary epidemiological data collected from SAC in Mangochi District, Lake Malawi, as published previously. Available diagnostic data by child, were converted into binary response infection profiles for 520 children, aged 6-15, across 12 sampled schools. Generalised additive models were then fitted to mono- and dual-infections. These were used to identify consistent population trends, finding the prevalence of IS significantly increased [p = 8.45e-4] up to 11 years of age then decreasing thereafter. A similar age-prevalence association was observed for co-infection [p = 7.81e-3]. By contrast, no clear age-infection pattern for UGS was found [p = 0.114]. Peak prevalence of Schistosoma infection typically occurs around adolescence; however, in this newly established IS outbreak with rising prevalence of UGS co-infections, the peak appears to occur earlier, around the age of 11 years. As the outbreak of IS fulminates, further temporal analysis of the age-relationship with Schistosoma infection is justified. This should refer to age-prevalence models which could better reveal newly emerging transmission trends and Schistosoma species dynamics. Dynamical modelling of infections, alongside malacological niche mapping, should be considered to guide future primary data collection and intervention programmes.

Biomphalaria pfeifferi (Gastropoda: Planorbidae) in Lake Malawi and Upper Shire River, Mangochi District, Malawi: Distribution, Genetic Diversity and Pre-Patent Schistosome Infections.

In November 2017, Biomphalaria pfeifferi, the key intermediate host for Schistosoma mansoni in Africa, was first reported in Lake Malawi, Mangochi District. Two subsequent malacological surveys in 2018 and 2019 confirmed its lacustrine presence, as well as its presence along the Upper Shire River. These surveys provided sufficient specimens for analyses of the genetic structure and a transmission assessment for intestinal schistosomiasis. A total of 76 collected snails were characterized by a DNA sequence analysis of a 650 bp fragment of the mitochondrial cytochrome oxidase subunit 1 (cox1); by size fractionation of six fluorescently labelled microsatellite loci (Bgµl16, Bgµl, Bpf8, rg6, U-7, and rg9);by denaturing PAGE; and by detection of pre-patent Schistosoma infection by real-time PCR with a TaqMan® probe. Five closely related cox1 haplotypes were identified, all present within a single location, with only one haplotype common across all the other locations sampled. No allelic size variation was detected with the microsatellites and all loci were monomorphic. Overall, the pre-patent prevalence of Schistosoma spp. was 31%, with infected snails found at several sampling locations. In this part of Lake Malawi, Bi. pfeifferi exhibits low genetic diversity and is clearly being exposed to the miracidia of S. mansoni, which is likely facilitating the autochthonous transmission of this parasite.

Detection of male genital schistosomiasis (MGS) by real-time TaqMan® PCR analysis of semen from fishermen along the southern shoreline of Lake Malawi.

Background: Male genital schistosomiasis (MGS) is an underappreciated complication of schistosomiasis, first described in 1911. However, its epidemiology, diagnostic testing and case management are not well understood in sub-Saharan Africa. To shed new light on MGS prevalence in Malawi, a longitudinal cohort study was conducted among adult fishermen along the southern shoreline of Lake Malawi using detection of schistosome DNA in participants' semen by real-time TaqMan® PCR analyses. Methods: Upon recruitment of 376 participants, 210 submitted urine samples and 114 semen samples for parasitological tests. Thereafter, the available semen samples were subsequently analysed by real-time TaqMan® PCR. Praziquantel (PZQ) treatment was provided to all participants with follow-ups attempted at 1, 3, 6 and 12-months' intervals. Results: At baseline, real-time PCR detected a higher MGS cohort prevalence of 26.6% (n = 64, Ct-value range: 18.9-37.4), compared to 10.4% by semen microscopy. In total, 21.9% of participants (n = 114) were detected with MGS either by semen microscopy and/or by real-time PCR. Subsequent analyses at 1-, 3-, 6- and 12-month follow-ups indicated variable detection dynamics. Conclusions: This first application of a molecular method, to detect MGS in sub-Saharan Africa, highlights the need for development of such molecular diagnostic tests which should be affordable and locally accessible. Our investigation also notes the persistence of MGS over a calendar year despite praziquantel treatment.

Prospective pilot study on the relationship between seminal HIV-1 shedding and genital schistosomiasis in men receiving antiretroviral therapy along Lake Malawi.

Male genital schistosomiasis (MGS) is hypothesized to increase seminal shedding of HIV-1. This prospective pilot study assessed seminal HIV-1 RNA shedding in men on long-term ART with and without a diagnosis of MGS. Study visits occurred at 0, 1, 3, 6 and 12 months. MGS was diagnosed by egg positivity on semen microscopy or PCR of seminal sediment. After optimization of the HIV-RNA assay, we examined 72 paired plasma and semen samples collected from 31 men (15 with and 16 without MGS) over 12 months. HIV-1 RNA was detected in 7/72 (9.7%) seminal samples and 25/72 (34.7%) plasma samples. When comparing sample pairs, 5/72 (6.9%) showed HIV-1 RNA detection only in the seminal sample. Overall, 3/31 (9.7%) participants, all with MGS, had detectable HIV-1 RNA in semen while plasma HIV-1 RNA was undetectable (< 22 copies/mL), with seminal levels ranging up to 400 copies/mL. Two participants showing HIV-1 RNA in seminal fluid from the MGS-negative group also had concomitant HIV-1 RNA detection in plasma. The findings suggest that MGS can be associated with low-level HIV-1 RNA shedding despite virologically suppressive ART. Further studies are warranted to confirm these observations and assess its implications.

Molecular Epidemiology and Assemblage Typing of Giardia duodenalis in School-Age Children Situated along the Southern Shoreline of Lake Malawi, Malawi.

Almost all human giardiasis infections are caused by Giardia duodenalis assemblages A and B. Differentiation between human infections with these assemblages, as well as between single-assemblage (A or B) and mixed-assemblage (A and B) infections, is therefore needed to better understand the pathological impact of infection with either, or both, assemblages. We assessed the prevalence of G. duodenalis assemblages A and B using 305 fecal samples provided by school-age children situated along the southern shoreline of Lake Malawi. Concurrently, intestinal pathology data were also collected to test for association(s) between assemblage infection status and intestinal health. Prevalence of G. duodenalis infection was 39.3% by real-time polymerase chain reaction. Of all identified infections, 32% were single G. duodenalis assemblage A and 32% were single G. duodenalis assemblage B, whereas 33% were mixed-assemblage infections. Fifteen unique G. duodenalis assemblage A and 13 unique G. duodenalis assemblage B ß-giardin haplotypes were identified. There was a positive association between single infection with G. duodenalis assemblage B and both self-reporting of abdominal pain (odds ratio [OR]: 3.05, P = 0.004) and self-reporting of diarrhea (OR: 3.1, P = 0.003). No association between single infection with assemblage A and any form of intestinal pathology was found. Additionally, there was a positive association between mixed-assemblage infections and self-reporting of abdominal pain (OR: 3.1, P = 0.002). Our study highlights the importance G. duodenalis assemblage typing and reaffirms the need for improved access to water, sanitation and hygiene infrastructure in rural areas of low- and middle-income countries.

Pathological Abnormalities Observed on Ultrasonography among Fishermen Associated with Male Genital Schistosomiasis (MGS) along the South Lake Malawi Shoreline in Mangochi District, Malawi.

Schistosome eggs cause granulomata and pathological abnormalities, detectable with non-invasive radiological techniques such as ultrasonography which could be useful in male genital schistosomiasis (MGS). As part of our novel MGS study among fishermen along Lake Malawi, we describe pathologies observed on ultrasonography and praziquantel (PZQ) treatment over time. Fishermen aged 18+ years were recruited, submitted urine and semen for parasitological and molecular testing, and thereafter, transabdominal pelvic and scrotal ultrasonography, assessing pathologies in the prostate, seminal vesicles, epididymis and testes. Standard PZQ treatment and follow-up invitation at 1-, 3-, 6- and 12-months' time-points were offered. A total of 130 recruited fishermen underwent ultrasonography at baseline (median age: 32.0 years); 27 (20.9%, n = 129) had S. haematobium eggs in urine (median: 1.0 egg/10 mL), 10 (12.3%, n = 81) in semen (defined as MGS, median: 2.9 eggs/mL ejaculate) and 16 (28.1%, n = 57) had a positive seminal Schistosoma real-time PCR. At baseline, 9 fishermen (6.9%, n = 130) had abnormalities, with 2 positive MGS having prostatic and testicular nodules. Fewer abnormalities were observed on follow-up. In conclusion, pathologies detected in male genitalia by ultrasonography can describe MGS morbidity in those with positive parasitological and molecular findings. Ultrasonography advances and accessibility in endemic areas can support monitoring of pathologies' resolution after treatment.

First Molecular Identification of Bulinus africanus in Lake Malawi Implicated in Transmitting Schistosoma Parasites.

The freshwater snail genus Bulinus plays a vital role in transmitting parasites of the Schistosoma haematobium group. A hybrid schistosome between S. haematobium and S. mattheei has been recently detected using DNA-based identification methods in school children along the Lake Malawi shoreline in Mangochi District. This finding raised the need for contemporary revaluation of local interactions between schistosomes and snails, with a particular focus on snail species within the Bulinus africanus group. In 2017 and 2018, malacological surveys sampled several freshwater sites in Mangochi District. Collected snails (n = 250) were characterised using cytochrome oxidase subunit 1 gene (cox1), with DNA barcoding of the 'Folmer' region and a rapid PCR-RFLP typing assay with double digestion with HaeIII and SacI restriction enzymes. DNA cox1 sequence analysis, with phylogenetic tree construction, suggested the presence of at least three B. africanus group taxa in Lake Malawi, B. globosus, alongside first reports of B. africanus and B. angolensis, which can be differentiated by PCR-RFLP methods. In addition, a total of 30 of the 106 B. africanus group snails (28.30%) were positive to the Schistosoma-specific screen using real-time PCR methods. This study provides new insight into the recent changes in the epidemiology of urogenital schistosomiasis as likely driven by a new diversity of B. africanus group snails within the Lake.

Male Genital Schistosomiasis Along the Shoreline of Lake Malawi: Baseline Prevalence and Associated Knowledge, Attitudes and Practices Among Local Fishermen in Mangochi District, Malawi.

Male genital schistosomiasis (MGS) is an often-overlooked chronic consequence of urogenital schistosomiasis (UGS) associated with Schistosoma haematobium eggs and associated pathologies in the genital system of afflicted men. Despite the first formal description of MGS in 1911 by Madden, its epidemiology, diagnostic testing and case management of today are not well-described. However, since several interactions between MGS and the Human Immunodeficiency Virus (HIV) are known, there is renewed public health interest in MGS across sub-Saharan Africa (SSA). To shed new light upon MGS in Malawi, a longitudinal cohort study was set up among fishermen along the southern shoreline of Lake Malawi in Mangochi District, Malawi, to document its prevalence and assess mens' knowledge, attitudes and practices (KAP). After providing informed written consent, fishermen (n = 376) aged 18+ years (median age: 30 years, range: 18-70 years) were recruited and submitted urine and semen for point-of-care (POC) field and laboratory diagnostic parasitological tests. Individual questionnaires were administered to assess their KAP, with praziquantel (PZQ) treatment provided to all participants. Baseline prevalence of MGS (S. haematobium eggs in semen) was 10.4% (n = 114, median: 5.0 eggs per ml, range: 0.1-30.0) while for UGS (S. haematobium eggs in urine) was 17.1% (n = 210, median: 2.3 eggs per 10 ml, range: 0.1-186.0) and 3.8% were positive by POC circulating cathodic antigen (POC-CCA), indicative of a Schistosoma mansoni infection. Just under 10% of participants reported having experienced symptoms associated with MGS, namely genital or coital pain, or haemospermia. A total of 61.7% reported previous difficulties in accessing PZQ therapy, with 34.8% having received PZQ therapy before. There was a significant correlation between MGS infection and the frequency of fishing in a week (rho = -0.25, n = 100, p = 0.01). In conclusion, MGS is prevalent among local fishermen yet knowledge of the disease is poor. We therefore call for improved availability and accessibility to MGS diagnostics, PZQ treatment within ongoing control interventions. This will improve the lives and reproductive health of men, their partners and communities in this shoreline environment of Lake Malawi.

An outbreak of intestinal schistosomiasis, alongside increasing urogenital schistosomiasis prevalence, in primary school children on the shoreline of Lake Malawi, Mangochi District, Malawi.

BACKGROUND: Intestinal schistosomiasis was not considered endemic in Lake Malawi until November 2017 when populations of Biomphalaria pfeifferi were first reported; in May 2018, emergence of intestinal schistosomiasis was confirmed. This emergence was in spite of ongoing control of urogenital schistosomiasis by preventive chemotherapy. Our current study sought to ascertain whether intestinal schistosomiasis is transitioning from emergence to outbreak, to judge if stepped-up control interventions are needed. METHODS: During late-May 2019, three cross-sectional surveys of primary school children for schistosomiasis were conducted using a combination of rapid diagnostic tests, parasitological examinations and applied morbidity-markers; 1) schistosomiasis dynamics were assessed at Samama (n = 80) and Mchoka (n = 80) schools, where Schistosoma mansoni was first reported, 2) occurrence of S. mansoni was investigated at two non-sampled schools, Mangochi Orphan Education and Training (MOET) (n = 60) and Koche (n = 60) schools, where B. pfeifferi was nearby, and 3) rapid mapping of schistosomiasis, and B. pfeifferi, conducted across a further 8 shoreline schools (n = 240). After data collection, univariate analyses and Chi-square testing were performed, followed by binary logistic regression using generalized linear models, to investigate epidemiological associations. RESULTS: In total, 520 children from 12 lakeshore primary schools were examined, mean prevalence of S. mansoni by 'positive' urine circulating cathodic antigen (CCA)-dipsticks was 31.5% (95% confidence interval [CI]: 27.5-35.5). Upon comparisons of infection prevalence in May 2018, significant increases at Samama (relative risk [RR] = 1.7, 95% CI: 1.4-2.2) and Mchoka (RR = 2.7, 95% CI: 1.7-4.3) schools were observed. Intestinal schistosomiasis was confirmed at MOET (18.3%) and Koche (35.0%) schools, and in all rapid mapping schools, ranging from 10.0 to 56.7%. Several populations of B. pfeifferi were confirmed, with two new eastern shoreline locations noted. Mean prevalence of urogenital schistosomiasis was 24.0% (95% CI: 20.3-27.7). CONCLUSIONS: We notify that intestinal schistosomiasis, once considered non-endemic in Lake Malawi, is now transitioning from emergence to outbreak. Once control interventions can resume after coronavirus disease 2019 (COVID-19) suspensions, we recommend stepped-up preventive chemotherapy, with increased community-access to treatments, alongside renewed efforts in appropriate environmental control.

Case Report: Highlighting Male Genital Schistosomiasis (MGS) in Fishermen from the Southwestern Shoreline of Lake Malawi, Mangochi District.

Urogenital schistosomiasis causes morbidity within the genitalia but is underreported and infrequently examined in men. To draw attention to male genital schistosomiasis (MGS), a longitudinal cohort study was conducted among fishermen along the southwestern shoreline of Lake Malawi. A case series of five participants is presented inclusive of questionnaire interviews, parasitological examinations, ultrasonography, and provision of a standard dose (40 mg/kg) of praziquantel (PZQ) treatment at baseline, 1-, 3-, 6-, and 12-month follow-up time points. Eggs of Schistosoma haematobium were observed in urine or semen across all time points; parasitological diagnostics were bolstered by real-time PCR for Schistosoma DNA in semen and by portable ultrasonography to document putative MGS-associated morbidity. We highlight the importance of developing standard diagnostic tests for MGS and increasing the accessibility of PZQ treatment to men, especially those in at-risk endemic areas.