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Adult schistosomes live in the blood vessels and cannot easily be sampled from humans, so archived miracidia larvae hatched from eggs expelled in feces or urine are commonly used for population genetic studies. Large collections of archived miracidia on FTA cards are now available through the Schistosomiasis Collection at the Natural History Museum (SCAN). Here we describe protocols for whole genome amplification of Schistosoma mansoni and Schistosome haematobium miracidia from these cards, as well as real time PCR quantification of amplified schistosome DNA. We used microgram quantities of DNA obtained for exome capture and sequencing of single miracidia, generating dense polymorphism data across the exome. These methods will facilitate the transition from population genetics, using limited numbers of markers to population genomics using genome-wide marker information, maximising the value of collections such as SCAN.
Endeavours to control urogenital schistosomiasis on Unguja Island (Zanzibar) have focused on school-aged children. To assess the impact of an associated health education campaign, the supervised use of the comic-strip medical booklet Juma na Kichocho by Class V pupils attending eighteen primary schools was investigated. A validated knowledge and attitudes questionnaire was completed at baseline and repeated one year later following the regular use of the booklet during the calendar year. A scoring system (ranging from 0.0 to 5.0) measured children’s understandings of schistosomiasis and malaria, with the latter being a neutral comparator against specific changes for schistosomiasis. In 2006, the average score from 751 children (328 boys and 423 girls) was 2.39 for schistosomiasis and 3.03 for malaria. One year later, the score was 2.43 for schistosomiasis and 2.70 for malaria from 779 children (351 boys and 428 girls). As might be expected, knowledge and attitudes scores for schistosomiasis increased (+0.05), but not as much as originally hoped, while the score for malaria decreased (−0.33). According to a Kolmogorov–Smirnov test, neither change was statistically significant. Analysis also revealed that 75% of school children misunderstood the importance of reinfection after treatment with praziquantel. These results are disappointing. They demonstrate that it is mistaken to assume that knowledge conveyed in child-friendly booklets will necessarily be interpreted, and acted upon, in the way intended. If long-term sustained behavioural change is to be achieved, health education materials need to engage more closely with local understandings and responses to urogenital schistosomiasis. This, in turn, needs to be part of the development of a more holistic, biosocial approach to the control of schistosomiasis.
Top-down biomedical interventions to control schistosomiasis in sub-Saharan Africa have had limited success, primarily because they fail to engage with the social, political, economic and ecological contexts in which they are delivered. Despite the call to foster community engagement and to adapt interventions to local circumstances, programmes have rarely embraced such an approach. This article outlines a community co-designed process, based upon Human-Centered Design, to demonstrate how this approach works in practice. It is based on initial work undertaken by social science researchers, public health practitioners and community members from the Zanzibar Islands, Tanzania, between November 2011 and December 2013. During the process, 32 community members participated in a qualitative and quantitative data-driven workshop where they interpreted data on local infections from S. haematobium and co-designed interventions with the assistance of a facilitator trained in the social sciences. These interventions included the implementation of novel school-based education and training, the identification of relevant safe play activities and events at local schools, the installation of community-designed urinals for boys and girls and the installation of community-designed laundry-washing platforms to reduce exposure to cercariae-contaminated fresh water. It is suggested that the a community co-designed process, drawing from Human-Centered Design principles and techniques, enables the development of more sustainable and effective interventions for the control of schistosomiasis.
Meta-analyses show that cognitive behaviour therapy for psychosis (CBT-P) improves distressing positive symptoms. However, it is a complex intervention involving a range of techniques. No previous study has assessed the delivery of the different elements of treatment and their effect on outcome. Our aim was to assess the differential effect of type of treatment delivered on the effectiveness of CBT-P, using novel statistical methodology.
The Psychological Prevention of Relapse in Psychosis (PRP) trial was a multi-centre randomized controlled trial (RCT) that compared CBT-P with treatment as usual (TAU). Therapy was manualized, and detailed evaluations of therapy delivery and client engagement were made. Follow-up assessments were made at 12 and 24 months. In a planned analysis, we applied principal stratification (involving structural equation modelling with finite mixtures) to estimate intention-to-treat (ITT) effects for subgroups of participants, defined by qualitative and quantitative differences in receipt of therapy, while maintaining the constraints of randomization.
Consistent delivery of full therapy, including specific cognitive and behavioural techniques, was associated with clinically and statistically significant increases in months in remission, and decreases in psychotic and affective symptoms. Delivery of partial therapy involving engagement and assessment was not effective.
Our analyses suggest that CBT-P is of significant benefit on multiple outcomes to patients able to engage in the full range of therapy procedures. The novel statistical methods illustrated in this report have general application to the evaluation of heterogeneity in the effects of treatment.
During the past decade, enormous technological developments have occurred in biology that have led to significant and revolutionary advances. New techniques of DNA cloning, restriction-enzyme analyses and nucleotide sequencing are providing a mass of data concerning the genomes of a wide variety of organisms. Such insights are having a great impact upon many areas of biological investigation, and would seem to be of considerable potential value for studies of taxonomy and population biology. The application of these new approaches to the characterization and identification of parasitic helminths has only recently begun, but they promise to become powerful additional tools for this purpose. Better methods of characterization are required for more precise definition of the parasites of man and domestic animals and for determining vectors and intermediate hosts as well as possible animal reservoir hosts. Moreover, a greater understanding of the genetic diversity of parasitic organisms is required since many helminths, which are morphologically similar, show marked differences in epidemiologically significant factors such as infectivity, pathogenicity, immunogenicity and drug sensitivity.
The recent implementation of mass drug administration (MDA) for control of uro-genital schistosomiasis has identified an urgent need for molecular markers to both directly monitor the impact of MDA, for example to distinguish re-infections from uncleared infections, as well as understand aspects of parasite reproduction and gene flow which might predict evolutionary change, such as the development and spread of drug resistance. We report the development of a novel microsatellite tool-kit allowing, for the first time, robust genetic analysis of individual S. haematobium larvae collected directly from infected human hosts. We genotyped the parasite populations of 47 children from 2 schools in the Ségou region of Mali, the first microsatellite study of this highly neglected parasite. There was only limited evidence of population subdivision between individual children or between the two schools, suggesting that few barriers to gene flow exist in this population. Complex relationships between parasite reproductive success, infection intensity and host age and gender were identified. Older children and boys harboured more diverse infections, as measured by the number of unique adult genotypes present. Individual parasite genotypes had variable reproductive success both across hosts, a pre-requisite for evolutionary selection, and, phenotypically, in hosts of different ages and genders. These data serve as a baseline against which to measure the effect of treatment on parasite population genetics in this region of Mali, and the tools developed are suitable to further investigate this important pathogen, and its close relatives, throughout their range.
All good quality trials of psychological interventions need to check formally that therapists have used the techniques prescribed in the published therapy manuals, and that the therapy has been carried out competently. This paper reviews methods of assessing adherence and competence used in recent large-scale trials of Cognitive Behaviour Therapy (CBT) for psychosis in the UK carried out by our research groups. A combination of the Cognitive Therapy Rating Scale and specific versions of the Cognitive Therapy for Psychosis Adherence Scales provides an optimal assessment of adherence and competence. Careful assessment of the competence and adherence can help identify the procedures actually carried out with individuals within trials. The basic use of such assessments is to provide an external check on treatment fidelity on a sample of sessions. Such assessment can also provide the first step towards moving research towards making sense of CBT for psychosis as a complex intervention and identifying which techniques work for which problems of people with psychosis, at which stages of disorder?
The current study considers the distribution of a small sample of 138 Bulinus snails, across 28 localities within eight Nigerian states. Snails were identified using a combination of molecular methods involving both DNA sequencing of a partial cytochrome oxidase subunit 1 (cox1) fragment and restriction profiles obtained from ribosomal internal transcribed spacer (its) amplicons. The results showed that the majority of Bulinus samples tested belonged to the species Bulinus truncatus while only two were Bulinus globosus. The use of RsaI restriction endonuclease to cleave the ribosomal its of Bulinus, as a method of species identification, was adopted for the majority of samples, this being a quicker and cheaper method better suited to small laboratory environments. Polymerase chain reaction (PCR) amplification of the schistosome Dra1 repeat within each of the collected Bulinus samples was employed to determine the extent and distribution of infected snails within the sample areas. Successful amplification of the Dra1 repeat demonstrated that 29.7% of snails were infected with schistosomes. Sequencing of the partial schistosome its from a small subset of snail samples suggested that some snails were either penetrated by both Schistosoma haematobium and Schistosoma bovis miracidia or hybrid miracidia formed from the two species.
Snails, provisionally identified as Bulinus tropicus, on the basis of chromosome number, egg protein profile, AcP and 118DH 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 schistosomcs 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.
Schistosoma haematobium and S. bovis are widespread schistosome species causing human and cattle schistosomiasis, respectively, in Africa. The sympatric occurrence of these two species and their ability to infect the same Bulinus intermediate snail hosts necessitates precise methods of identification of the larval stages. A rapid diagnostic ‘mulitplex’ one-step polymerase chain reaction protocol (RD-PCR) was developed using cytochrome oxidase subunit 1 (COX1) mitochondrial DNA (mtDNA) to discriminate between S. haematobium and S. bovis. A single forward primer and two species-specific reverse primers were used to produce a polymerase chain reaction (PCR) fragment of 306 bp and 543 bp for S. bovis and S. haematobium, respectively. Serial dilutions were carried out on various lifecycle stages and species combinations to test the sensitivity and specificity of the primers. This RD-PCR proved highly sensitive, detecting a single larval stage and as little as 0.78 ng of genomic DNA (gDNA) from an adult schistosome, providing a cost-effective, rapid and robust molecular tool for high-throughput screening of S. haematobium and S. bovis populations. In areas where human and cattle schistosomiasis overlap and are transmitted in close proximity, this mitochondrial assay will be a valuable identification tool for epidemiological studies, especially when used in conjunction with other nuclear diagnostic markers.
This review considers the current status of urinary schistosomiasis, caused by infection with Schistosoma haematobium, and argues that greater research effort and focus are needed to improve understanding of this neglected tropical disease (NTD). The inappropriateness of relying solely on data concerning the much more extensively studied intestinal form of schistosomiasis caused by S. mansoni is highlighted. The current lack of genome and transcriptome information for S. haematobium is directly hindering further targeted research and must be quickly rectified. Recent molecular phylogenies caution the expectation of similarities between schistosome species and highlight the close relationships of species within the S. haematobium group. Treatment, current and prospective drugs and vaccines, together with diagnosis are considered, highlighting the differences associated with urinary schistosomiasis. This infection has a significant and specific impact on the urino-genital system and has a strong association with bladder cancer, leading to severe and chronic morbidity. There is a clear need for new clinical initiatives in this area to better quantify the disease burden. Furthermore, emerging associations with HIV and other pathogens need to be closely monitored. Research is urgently needed to improve current knowledge in order to develop the next generation of control tools.
Representative samples of Ugandan Schistosoma mansoni from Lake Albert and Lake Victoria were examined using DNA barcoding, sequence analysis of two partially overlapping regions – ASMIT (396 bp) & MORGAN (617 bp) – of the mitochondrial cytochrome oxidase subunit I (cox1). The Victorian sample exhibited greater nucleotide diversity, 1·4% vs. 1·0%, and a significant population partition appeared as barcodes did not cross-over between lakes. With one exception, Lake Albert populations were more mixed by sampled location, while those from Lake Victoria appeared more secluded. Using statistical parsimony, barcode ASMIT 1 was putatively ancestral to all others and analysis of MORGAN cox1 confirmed population diversity. All samples fell into two of five well-resolved lineages; sub-lineages therein broadly partitioning by lake. It seems that barcode ASMIT 1 (and close variants) was likely widely dispersed throughout the Nilotic environment but later diversified in situ, and in parallel, within Lake Albert and Lake Victoria. The genetic uniformity of Ugandan S. mansoni can no longer be assumed, which might better explain known epidemiological heterogeneities. While it appears plausible that locally evolved heritable traits could spread through most of the Lake Albert populations, it seems unlikely they could quickly homogenise into Lake Victoria or amongst populations therein.
Water soluble extracts of 3131 adult specimens of Dicrocoelium dendriticum from cattle, sheep and goats, mainly from León province, were analysed by isoelectric focusing in thin-layer polyacrylamide gels. Activity of the following enzymes was studied: lactate dehydrogenase (LDH, EC 126.96.36.199), glucose phosphate isomerase (GPI, EC 188.8.131.52), phosphoglucomutase (PGM, EC 184.108.40.206), acid phosphatase (AcP, EC 220.127.116.11), α-glycerophosphate dehydrogenase (α-GPDH, EC 18.104.22.168), hydroxybutyrate dehydrogenase (HBDH, EC 22.214.171.124) and malate dehydrogenase (MDH, 126.96.36.199). Five distinct enzyme types were recognized for LDH (pH range 6.30–7.13), GPI (pH 6.13–6.80) and PGM (pH 6.20–6.60) whereas AcP showed three different patterns (pH 5.70–5.92). Weak and diffuse activity was detected for MDH (pH 4.8–6.2) and no activity was observed for α-GPDH and HBDH. In general, little phenotypic variation was observed between worms recovered from a single host, between those from hosts of the same species and between those from hosts of different species, although some enzyme types were found in some animals but not others. Nevertheless, it must be taken into account that most parasites came from sheep and also from a relatively small area in north-west Spain.
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 (6O%). 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.
A total of 358 cattle was examined for schistosome infection in Zambian slaughterhouses. A total of 542 worms collected from 104 infected individuals was examined for glucose-6-phosphate dehydrogenase and phosphoglucomutase using isoelectric focusing. The overall prevalence of infection was 51%. Ninety three percent of the infected animals had less than 100 worm pairs in the mesenteric veins. Schistosoma mattheei was the predominant species (75%); S. leiperi (12%) and S. margrebowiei (2%) were also identified. The remaining 11% of the worms showed one of two distinct heterozygote patterns. Pattern A is identical to that of a laboratory-produced Fl S. mattheei × S. haematobium hybrid, but could also represent a S. mattheei × S. leiperi hybrid. Further studies are required to elucidate the origins of pattern B.
Schistosoma curassoni has been recovered from cattle in northern Nigeria. Rectal scrapings of 90 cows slaughtered at the Kano abattoir, Kano, Nigeria during March and April 1986 revealed a prevalence of 7·8% S. bovis and 2·2% S. curassoni. Further examination of the mesenteric and rectal veins of 502 cows showed that the overall prevalence of schistosomiasis was 31·1%. Local Bulinus globosus were infected successfully in the laboratory with S. bovis miracidia.
Variability of Schistosoma intercalatum eggs in shape and size, and their similarity to those of S. haematobium presented a problem of species identification when egg morphology was the diagnostic criterion used in a study of human schistosomiasis conducted on São Tomé and Principe. More than 2500 egg measurements were obtained by light micoscopy to gather data relating to size variability of S. intercalatum eggs, to evaluate whether factors such as age of host, sex of host and intensity of infection are correlated with variability, and the data were compared with previously published measurements on different isolates and strains of S. intercalatum: the range in length (104–203 μm) embraces most of the measurements reported in other studies of S. intercalatum eggs. There was no correlation either between age and sex of the host, or intensity of infection with variability of egg size. Comparison between measurements of the eggs of S. haematobium, S. intercalatum and S. bovis eggs are presented.
An average of 11% of adult Schistosoma curassoni were recovered from 200 albino mice which had been infected subcutaneously with 150–250 cercariae. Worms were primarily found in the portal veins. The average number of intrauterine eggs per female during the first 100 days p.i. was 13 and the average number of eggs produced per female worm was 103 per day for the first 60 days post oviposition. The majority of eggs were recovered from the liver (98·3%). The oograms were determined until day 95 p.i. For screening of antischistosomal drugs it is recommended not to use infections older than 60 d.p.i.