This study documented the population dynamics of Biomphalaria and associated natural infections with digenetic trematodes, along the shores of Lake Albert and Lake Victoria, recording local physicochemical factors. Over a two-and-a-half-year study period with monthly sampling, physicochemical factors were measured at 12 survey sites and all freshwater snails were collected. Retained Biomphalaria were subsequently monitored in laboratory aquaria for shedding trematode cercariae, which were classified as either human infective (Schistosoma mansoni) or nonhuman infective. The population dynamics of Biomphalaria differed by location and by lake and had positive relationship with pH (P
BACKGROUND: Vector control is an effective way of reducing malaria transmission. The main vector control methods include the use of insecticide-treated bed nets and indoor residual spraying (IRS). Both interventions rely on the continuing susceptibility of Anopheles to a limited number of insecticides. However, insecticide resistance, in particular pyrethroid-DDT cross-resistance, is a challenge facing malaria vector control in Africa because pyrethroids represent the only class of insecticides approved for treating bed nets and DDT is commonly used for IRS. Here baseline data are presented on the insecticide susceptibility levels of malaria vectors prior to The Gambian indoor residual spraying intervention programme. METHODS: Anopheles larvae were collected from six malaria surveillance sites (Brikama, Essau, Farafenni, Mansakonko, Kuntaur and Basse) established by the National Malaria Control Programme and the UK Medical Research Council Laboratories in The Gambia. The mosquitoes were reared to adulthood and identified using morphological keys and a species-specific polymerase chain reaction assay. Two- to three-day old adult female mosquitoes were tested for susceptibility to permethrin, deltamethrin and DDT using standard WHO protocols, insecticide susceptibility test kits and treated papers. RESULTS: All Anopheles mosquitoes tested belonged to the Anopheles gambiae complex. Anopheles arabiensis was predominant (54.1%), followed by An. gambiae s.s. (26.1%) and Anopheles melas (19.8%). Anopheles gambiae s.s. and An. arabiensis were found at all six sites. Anopheles melas was recorded only at Brikama. Mosquitoes from two of the six sites (Brikama and Basse) were fully susceptible to all three insecticides tested. However, DDT resistance was found in An. gambiae from Essau where the 24 hours post-exposure mortality was
Howell A, Mugisha L, Davies J, LaCourse EJ, Claridge J, Williams DJ, Kelly-Hope L, Betson M, Kabatereine NB, Stothard JR (2012) Bovine fasciolosis at increasing altitudes: parasitological and malacological sampling on the slopes of Mount Elgon, Uganda., Parasites & vectors 5
BACKGROUND: To clarify the extent and putative transmission zone of bovine fasciolosis on the slopes of Mount Elgon, Uganda, conjoint parasitological and malacological surveys, inclusive of inspection of animals at slaughter, were undertaken at increasing altitudes. RESULTS: A total of 239 cattle were sampled across eight locations ranging in elevation from 1112-2072 m. Faecal material was examined for presence of Fasciola eggs and sera were tested by ELISA for antibodies against Fasciola antigens. Bolstering this, 38 cattle at slaughter from 2 abattoir sites at 1150 m and 1947 m were inspected; in addition, wild buffalo stool (n=10) opportunistically picked within Mount Elgon National Park (MENP) at 3640 m was examined. By faecal egg detection, prevalence of Fasciola gigantica at low (1500 m) altitude sites was 43.7% (95% CI 35.4-52.2) and 1.1% (95% CI 0.0-6.0), respectively, while by ELISA was much higher, low altitude--77.9% (95% CI 69.7-85.4) and high altitude--64.5% (95% CI 51.3-76.3). The decline in prevalence with increasing altitude was corroborated by abattoir sampling. Thirty seven aquatic habitats, ranging from 1139-3937 m in altitude were inspected for freshwater snails, 12 of which were within MENP. At lower altitudes, Lymnaea (Radix) natalensis was common, and often abundant, but at higher altitudes became much rarer ceasing to be found above 1800 m. On the other hand, Lymnaea (Galba) truncatula was found only at altitudes above 3000 m and within MENP alone. The snail identifications were confirmed by DNA analysis of the ribosomal 18S gene. CONCLUSIONS: Active infections of F. gigantica in cattle are common in lower altitude settings but appear to diminish with increasing elevation. This is likely due to a growing paucity of intermediate hosts, specifically populations of L. natalensis for which a natural boundary of 1800 m appeared. Although F. hepatica was not encountered, the presence of several populations of L. truncatula at elevations over 3000 m point towards a potential transmission zone within MENP should this parasite be introduced.
BACKGROUND: The roundworm Ascaris lumbricoides infects 0.8 billion people worldwide, and Ascaris suum infects innumerable pigs across the globe. The extent of natural cross-transmission of Ascaris between pig and human hosts in different geographical settings is unknown, warranting investigation. METHODS: Adult Ascaris organisms were obtained from humans and pigs in Europe, Africa, Asia, and Latin America. Barcodes were assigned to 536 parasites on the basis of sequence analysis of the mitochondrial cytochrome c oxidase I gene. Genotyping of 410 worms was also conducted using a panel of microsatellite markers. Phylogenetic, population genetic, and Bayesian assignment methods were used for analysis. RESULTS: There was marked genetic segregation between worms originating from human hosts and those originating from pig hosts. However, human Ascaris infections in Europe were of pig origin, and there was evidence of cross-transmission between humans and pigs in Africa. Significant genetic differentiation exists between parasite populations from different countries, villages, and hosts. CONCLUSIONS: In conducting an analysis of variation within Ascaris populations from pig and human hosts across the globe, we demonstrate that cross-transmission takes place in developing and developed countries, contingent upon epidemiological potential and local phylogeography. Our results provide novel insights into the transmission dynamics and speciation of Ascaris worms from humans and pigs that are of importance for control programs.
Frasa MA, Maximiano FC, Smolarczyk K, Francis RE, Betson ME, Lozano E, Goldenring J, Seabra MC, Rak A, Ahmadian MR, Braga VM (2010) Armus is a Rac1 effector that inactivates Rab7 and regulates E-cadherin degradation., Current biology : CB 20 (3) pp. 198-208
BACKGROUND: Cell-cell adhesion and intracellular trafficking are regulated by signaling pathways from small GTPases of the Rho, Arf, and Rab subfamilies. How signaling from distinct small GTPases are integrated in a given process is poorly understood. RESULTS: We find that a TBC/RabGAP protein, Armus, integrates signaling between Arf6, Rac1, and Rab7 during junction disassembly. Armus binds specifically to activated Rac1 and its C-terminal TBC/RabGAP domain inactivates Rab7. Thus, Armus is a novel Rac1 effector and a bona fide GAP for Rab7 in vitro and in vivo, a unique and previously unreported combination. Arf6 activation efficiently disrupts cell-cell contacts and is known to activate Rac1 and Rab7. Arf6-induced E-cadherin degradation is efficiently blocked by expression of Armus C-terminal domain or after Armus RNAi. Coexpression of Arf6 with dominant-negative Rab7 or Rac1 also inhibits junction disassembly. Importantly, Armus RabGAP expression also prevents EGF-induced scattering in keratinocytes, a process shown here to require Arf6, Rac1, and Rab7 function. To our knowledge, this is the first report to demonstrate a molecular and functional link between Rac1 and Rab7. CONCLUSIONS: Our data indicate that active Rac1 recruits Armus to locally inactivate Rab7 and facilitate E-cadherin degradation in lysosomes. Thus, the integration of Rac1 and Rab7 activities by Armus provides an important regulatory node for E-cadherin turnover and stability of cell-cell contacts.
Pinot de Moira A, Sousa-Figueiredo JC, Jones FM, Fitzsimmons CM, Betson M, Kabatereine NB, Stothard JR, Dunne DW (2013) Schistosoma mansoni infection in preschool-aged children: development of immunoglobulin E and immunoglobulin G4 responses to parasite allergen-like proteins., The Journal of infectious diseases 207 (2) pp. 362-366
Specific immunoglobulin E (IgE) responses are upregulated during chronic schistosome infection and during allergy. These responses are tightly regulated during schistosomiasis. We have previously shown that IgE regulation depends on the extent and length of exposure to individual parasite allergen-like proteins. Here we compare the development of IgE and immunoglobulin G4 (IgG(4)) responses to the differentially expressed allergen-like proteins SmTAL1 and SmTAL2 among preschool-aged children from 2 villages with different levels of Schistosoma mansoni transmission. We found a lack of SmTAL1 responsiveness among all children, but evidence for IgG(4)-dependent IgE-SmTAL2 desensitization in both villages, occurring earlier among children from the village where the level of transmission was greater. Findings provide insights into the development and regulation of allergic-type immune responses.
Green HK, Sousa-Figueiredo JC, Basáñez MG, Betson M, Kabatereine NB, Fenwick A, Stothard JR (2011) Anaemia in Ugandan preschool-aged children: the relative contribution of intestinal parasites and malaria., Parasitology 138 (12) pp. 1534-1545
Anaemia is a severe public health issue among African preschool-aged children, yet little effective progress has been made towards its amelioration, in part due to difficulties in unravelling its complex, multifactorial aetiology. To determine the current anaemia situation and assess the relative contribution of malaria, intestinal schistosomiasis and infection with soil-transmitted helminths, two separate cross-sectional epidemiological surveys were carried out in Uganda including 573 and 455 preschool-aged children (d6 years) living along the shores of Lake Albert and on the islands in Lake Victoria, respectively. Anaemia was found to be a severe public health problem in Lake Albert, affecting 68·9% of children (ninety-five percent confidence intervals (95% CI) 64·9-72·7%), a statistically significant higher prevalence relative to the 27·3% detected in Lake Victoria (95% CI: 23·3-31·7%). After multivariate analysis (controlling for sex and age of the child), the only factor found to be significantly associated with increased odds of anaemia in both lake systems was malaria (Lake Albert, odds ratio (OR)=2·1, 95% CI: 1·4-3·2; Lake Victoria, OR=1·9, 95% CI: 1·2-2·9). Thus intervention strategies primarily focusing on very young children and combating malaria appear to represent the most appropriate use of human and financial resources for the prevention of anaemia in this age group and area. Looking to the future, these activities could be further emphasised within the National Child Health Days(PLUS) agenda.
The complexity and connectedness of eco-social processes have major influence on the emergence and spread of infectious diseases amongst humans and animals. The disciplinary nature of most research activity has made it difficult to improve our understanding of interactions and feedback loops within the relevant systems. Influenced by the One Health approach, increasing efforts have recently been made to address this knowledge gap. Disease emergence and spread is strongly influenced by host density and contact structures, pathogen characteristics and pathogen population and molecular evolutionary dynamics in different host species, and host response to infection. All these mechanisms are strongly influenced by eco-social processes, such as globalisation and urbanisation, which lead to changes in global ecosystem dynamics, including patterns of mobility, human population density and contact structures, and food production and consumption. An improved understanding of epidemiological and eco-social processes, including their interdependence, will be essential to be able to manage diseases in these circumstances. The interfaces between wild animals, domestic animals and humans need to be examined to identify the main risk pathways and put in place appropriate mitigation. Some recent examples of emerging infectious disease are described to illustrate eco-social processes that are influencing disease emergence and spread.
Haygarth PM, Apsimon H, Betson M, Harris D, Hodgkinson R, Withers PJ (2009) Mitigating diffuse phosphorus transfer from agriculture according to cost and efficiency., Journal of environmental quality 38 (5) pp. 2012-2022
Potential options for mitigating phosphorus (P) transfer from agriculture to water in England and Wales (E&W) were collated across a range of farm systems to assess their potential effectiveness in reducing mass of P transferred and potential cost (pounds sterling [ pound]) to the farming industry. A simple model framework (called PEASE) incorporating a number of assumptions was used to identify 15 methods for mitigating inputs of P to agricultural systems, 19 methods for preventing mobilization of P, and six methods for controlling the transport of P to streams. The scope for largest reductions in P inputs was to grassland and horticulture. Potential reductions in P mobilization were up to 1.2 kg P ha(-1). Reductions in P transfer associated with transport mitigation were larger than those associated with input and mobilization methods (up to 2.2 kg P ha(-1)). The largest estimated reductions were achieved by installing buffer zones and constructed wetlands, the former being very cost effective ( pound3-5 kg(-1) P saved). Plots of cost curves helped identify where the combined and cumulative P transfer reductions were attainable; these were approximately 0.2 kg ha(-1) for uplands, 0.6 kg ha(-1) for outdoor pigs, 0.9 kg ha(-1) for intensive dairy, and 2.2 kg ha(-1) for arable examples. We concluded that established catchment-scale evidence for mitigation is sparse, especially for specific farm systems in E&W. Sensitivities and uncertainties in the approach, especially associated with expert coefficients, are noted. This approach is nonetheless considered useful for prioritizing where and how best options might be most effectively targeted for least cost but greatest benefit.
Betson M, Halstead FD, Nejsum P, Imison E, Khamis IS, Sousa-Figueiredo JC, Rollinson D, Stothard JR (2011) A molecular epidemiological investigation of Ascaris on Unguja, Zanzibar using isoenyzme analysis, DNA barcoding and microsatellite DNA profiling., Transactions of the Royal Society of Tropical Medicine and Hygiene 105 (7) pp. 370-379
Ascariasis is of public health importance on the islands of Zanzibar (Unguja and Pemba). To shed light on the molecular epidemiology of this parasite, 68 Ascaris worms, obtained from 14 individuals in four Ungujan villages, were examined by isoenzyme analysis (ISA), DNA barcoding and microsatellite DNA profiling. ISA revealed genetic variation, which was confirmed by DNA barcoding. Nineteen worms recovered from individuals in Uganda were included for comparison. Sixteen unique DNA barcodes were identified, 15 on Unguja and three in Uganda with two shared between. These two barcodes were found in all four Ungujan villages. Worms from Tumbatu-Jongowe, an isolated village on an islet off Unguja, seemed particularly diverse. Within our barcodes, three exact matches were found with Chinese Ascaris retrieved from pigs, which is perhaps surprising given the present rarity of these animals on Unguja. Microsatellite profiling and population genetic analysis revealed further genetic diversity within our samples although population sub-structuring within Unguja was minor in comparison to that between Unguja and Uganda. As African Ascaris has not been subjected to detailed molecular scrutiny, this new diversity represents an important piece in its evolutionary jigsaw and such population markers are informative in monitoring worm dynamics during ongoing control.
Verdier V, Johndrow JE, Betson M, Chen GC, Hughes DA, Parkhurst SM, Settleman J (2006) Drosophila Rho-kinase (DRok) is required for tissue morphogenesis in diverse compartments of the egg chamber during oogenesis., Developmental biology 297 (2) pp. 417-432
The Rho-kinases are widely utilized downstream targets of the activated Rho GTPase that have been directly implicated in many aspects of Rho-dependent effects on F-actin assembly, acto-myosin contractility, and microtubule stability, and consequently play an essential role in regulating cell shape, migration, polarity, and division. We have determined that the single closely related Drosophila Rho-kinase ortholog, DRok, is required for several aspects of oogenesis, including maintaining the integrity of the oocyte cortex, actin-mediated tethering of nurse cell nuclei, "dumping" of nurse cell contents into the oocyte, establishment of oocyte polarity, and the trafficking of oocyte yolk granules. These defects are associated with abnormalities in DRok-dependent actin dynamics and appear to be mediated by multiple downstream effectors of activated DRok that have previously been implicated in oogenesis. DRok regulates at least one of these targets, the membrane cytoskeletal cross-linker DMoesin, via a direct phosphorylation that is required to promote localization of DMoesin to the oocyte cortex. The collective oogenesis defects associated with DRok deficiency reveal its essential role in multiple aspects of proper oocyte formation and suggest that DRok defines a novel class of oogenesis determinants that function as key regulators of several distinct actin-dependent processes required for proper tissue morphogenesis.
Zhang J, Betson M, Erasmus J, Zeikos K, Bailly M, Cramer LP, Braga VM (2005) Actin at cell-cell junctions is composed of two dynamic and functional populations., Journal of cell science 118 (Pt 23) pp. 5549-5562
The ability of epithelial cells to polarize requires cell-cell adhesion mediated by cadherin receptors. During cell-cell contact, the mechanism via which a flat, spread cell shape is changed into a tall, cuboidal epithelial morphology is not known. We found that cadherin-dependent adhesion modulates actin dynamics by triggering changes in actin organization both locally at junctions and within the rest of the cell. Upon induction of cell-cell contacts, two spatial actin populations are distinguishable: junctional actin and peripheral thin bundles. With time, the relative position of these two populations changes and becomes indistinguishable to form a cortical actin ring that is characteristic of mature, fully polarized epithelial cells. Junctional actin and thin actin bundles differ in their actin dynamics and mechanism of formation, and interestingly, have distinct roles during epithelial polarization. Whereas junctional actin stabilizes clustered cadherin receptors at cell-cell contacts, contraction of peripheral actin bundle is essential for an increase in the maximum height at the lateral domain during polarization (cuboidal morphology). Thus, both junctional actin and thin bundles are necessary, and cooperate with each other to generate a polarized epithelial morphology.
Sousa-Figueiredo JC, Pleasant J, Day M, Betson M, Rollinson D, Montresor A, Kazibwe F, Kabatereine NB, Stothard JR (2010) Treatment of intestinal schistosomiasis in Ugandan preschool children: best diagnosis, treatment efficacy and side-effects, and an extended praziquantel dosing pole., International health 2 (2) pp. 103-113
The Ugandan national control programme for schistosomiasis has no clear policy for inclusion of preschool-children (=5 years old) children. To re-balance this health inequality, we sought to identify best diagnosis of intestinal schistosomiasis, observe treatment safety and efficacy of praziquantel (PZQ), and extend the current WHO dose pole for chemotherapy. We examined and treated 363 preschool children from shoreline villages of Lakes Albert and Victoria, and found that 62.3% (CI(95) 57.1-67.3) of the children were confirmed to have intestinal schistosomiasis. One day after treatment, children were reported as having headaches (3.6%), vomiting (9.4%), diarrhoea (10.9%) and urticaria/rash (8.9%) with amelioration at 21-day follow-up, where the parasitological cure rate was found to be 100.0%. Height and weight data were collected from a further 3303 preschool children to establish and validate an extended PZQ dose pole that now includes two new height-intervals: 60-84 cm for one-half tablet and 84-99 cm for three-quarter tablet divisions; which would result in 97.6% of children receiving an acceptable dose (30-60 mg/kg). To conclude, preschool children in lakeshore communities of Uganda are at significant risk of intestinal schistosomiasis; we now strongly advocate for their immediate inclusion within the national control programme to eliminate this health inequity.
Betson M, Sousa-Figueiredo JC, Atuhaire A, Arinaitwe M, Adriko M, Mwesigwa G, Nabonge J, Kabatereine NB, Sutherland CJ, Stothard JR (2014) Detection of persistent Plasmodium spp. infections in Ugandan children after artemether-lumefantrine treatment., Parasitology 141 (14) pp. 1880-1890
During a longitudinal study investigating the dynamics of malaria in Ugandan lakeshore communities, a consistently high malaria prevalence was observed in young children despite regular treatment. To explore the short-term performance of artemether-lumefantrine (AL), a pilot investigation into parasite carriage after treatment(s) was conducted in Bukoba village. A total of 163 children (aged 2-7 years) with a positive blood film and rapid antigen test were treated with AL; only 8.7% of these had elevated axillary temperatures. On day 7 and then on day 17, 40 children (26.3%) and 33 (22.3%) were positive by microscopy, respectively. Real-time PCR analysis demonstrated that multi-species Plasmodium infections were common at baseline, with 41.1% of children positive for Plasmodium falciparum/Plasmodium malariae, 9.2% for P. falciparum/ Plasmodium ovale spp. and 8.0% for all three species. Moreover, on day 17, 39.9% of children infected with falciparum malaria at baseline were again positive for the same species, and 9.2% of those infected with P. malariae at baseline were positive for P. malariae. Here, chronic multi-species malaria infections persisted in children after AL treatment(s). Better point-of-care diagnostics for non-falciparum infections are needed, as well as further investigation of AL performance in asymptomatic individuals.
Hawash MB, Betson M, Al-Jubury A, Ketzis J, LeeWillingham A, Bertelsen MF, Cooper PJ, Littlewood DT, Zhu XQ, Nejsum P (2016) Whipworms in humans and pigs: origins and demography., Parasit Vectors 9
BACKGROUND: Trichuris suis and T. trichiura are two different whipworm species that infect pigs and humans, respectively. T. suis is found in pigs worldwide while T. trichiura is responsible for nearly 460 million infections in people, mainly in areas of poor sanitation in tropical and subtropical areas. The evolutionary relationship and the historical factors responsible for this worldwide distribution are poorly understood. In this study, we aimed to reconstruct the demographic history of Trichuris in humans and pigs, the evolutionary origin of Trichuris in these hosts and factors responsible for parasite dispersal globally. METHODS: Parts of the mitochondrial nad1 and rrnL genes were sequenced followed by population genetic and phylogenetic analyses. Populations of Trichuris examined were recovered from humans (n = 31), pigs (n = 58) and non-human primates (n = 49) in different countries on different continents, namely Denmark, USA, Uganda, Ecuador, China and St. Kitts (Caribbean). Additional sequences available from GenBank were incorporated into the analyses. RESULTS: We found no differentiation between human-derived Trichuris in Uganda and the majority of the Trichuris samples from non-human primates suggesting a common African origin of the parasite, which then was transmitted to Asia and further to South America. On the other hand, there was no differentiation between pig-derived Trichuris from Europe and the New World suggesting dispersal relates to human activities by transporting pigs and their parasites through colonisation and trade. Evidence for recent pig transport from China to Ecuador and from Europe to Uganda was also observed from their parasites. In contrast, there was high genetic differentiation between the pig Trichuris in Denmark and China in concordance with the host genetics. CONCLUSIONS: We found evidence for an African origin of T. trichiura which were then transmitted with human ancestors to Asia and further to South America. A host shift to pigs may have occurred in Asia from where T. suis seems to have been transmitted globally by a combination of natural host dispersal and anthropogenic factors.
Kane RA, Stothard JR, Rollinson D, Leclipteux T, Evraerts J, Standley CJ, Allan F, Betson M, Kaba R, Mertens P, Laurent T (2013) Detection and quantification of schistosome DNA in freshwater snails using either fluorescent probes in real-time PCR or oligochromatographic dipstick assays targeting the ribosomal intergenic spacer., Acta tropica 128 (2) pp. 241-249
Several DNA probes were designed for use in real-time polymerase chain reaction (PCR) assays to target sequence variation within the ribosomal intergenic spacer (IGS) of schistosomes. A sub-section of the IGS (
Nejsum P, Bertelsen MF, Betson M, Stothard JR, Murrell KD (2010) Molecular evidence for sustained transmission of zoonotic Ascaris suum among zoo chimpanzees (Pan troglodytes)., Veterinary parasitology 171 (3-4) pp. 273-276
Chimpanzees in the Copenhagen Zoo frequently excrete ascarid worms onto the cage floor in spite of a regular anthelmintic treatment program. Previously it had been shown that the source of the infections was of pig origin. However, it was unknown whether the recurrence of the infection was due to reintroduction of eggs from an external source or to a sustained transmission cycle within the zoo. We found that isolated eggs were able to embryonate to the infective J3 stage and PCR-RFLP analysis on the ITS region amplified from single embryonated eggs suggest these to be Ascaris suum. In addition, sequence analysis of the cox1 gene ('barcoding') on expelled worms followed by cluster analysis revealed that the chimpanzees are infected with pig A. suum which now, in spite of control efforts, has stabilized into a permanent transmission cycle in the zoo's chimpanzee troop.
Seto EY, Sousa-Figueiredo JC, Betson M, Byalero C, Kabatereine NB, Stothard JR (2012) Patterns of intestinal schistosomiasis among mothers and young children from Lake Albert, Uganda: water contact and social networks inferred from wearable global positioning system dataloggers., Geospatial health 7 (1) pp. 1-13
The establishment of a national control programme (NCP) in Uganda has led to routine treatment of intestinal schistosomiasis with praziquantel in the communities along Lake Albert. However, because regular water contact remains a way of life for these populations, re-infection continues to mitigate the sustainability of the chemotherapy-based programme. A six-month longitudinal study was conducted in one Lake Albert community with the aim of characterizing water contact exposure and infection among mothers and their young preschool-aged children as the latter are not yet formally included within the NCP. At baseline the cohort of 37 mothers, 36 preschool-aged children had infection prevalences of 62% and 67%, respectively, which diminished to 20% and 29%, respectively, at the 6-month post-treatment follow-up. The subjects wore global positioning system (GPS) datalogging devices over a 3-day period shortly after baseline, allowing for the estimation of time spent at the lakeshore as an exposure metric, which was found to be associated with prevalence at follow-up (OR = 2.1, P = 0.01 for both mothers and young children and odds ratio (OR) = 4.4, P = 0.01 for young children alone). A social network of interpersonal interactions was also derived from the GPS data, and the exposures were positively associated both with the number and duration of peer interaction, suggesting the importance of socio-cultural factors associated with water contact behaviour. The findings illustrate reduction in both prevalence and intensity of infection in this community after treatment as well as remarkably high rates of water contact exposure and re-infection, particularly among younger children. We believe that this should now be formally considered within NCP, which may benefit from more in-depth ethnographic exploration of factors related to water contact as this should provide new opportunities for sustaining control.
Betson M, Nejsum P, Llewellyn-Hughes J, Griffin C, Atuhaire A, Arinaitwe M, Adriko M, Ruggiana A, Turyakira G, Kabatereine NB, Stothard JR (2012) Genetic diversity of Ascaris in southwestern Uganda., Transactions of the Royal Society of Tropical Medicine and Hygiene 106 (2) pp. 75-83
Despite the common occurrence of ascariasis in southwestern Uganda, helminth control in the region has been limited. To gain further insights into the genetic diversity of Ascaris in this area, a parasitological survey in mothers (n=41) and children (n=74) living in two villages, Habutobere and Musezero, was carried out. Adult Ascaris worms were collected from infected individuals by chemo-expulsion using pyrantel pamoate treatment. Genetic diversity within these worms was assessed by inspection of DNA sequence variation in a mitochondrial marker and length polymorphism at microsatellite loci. Overall prevalence of ascariasis was 42.5% in mothers and 30.4% in their children and a total of 98 worms was examined from 18 hosts. Sequence analysis of a portion of the mitochondrial cytochrome c oxidase subunit 1 gene revealed 19 different haplotypes, 13 of which had not been previously encountered. Microsatellite analysis using eight loci provided evidence for high gene flow between worm populations from the two villages but comparing these worms with others obtained in a prior study on Unguja, Zanzibar, confirmed little genetic exchange and mixing of worm populations between the two areas. By adding to our understanding of the genetic diversity of Ascaris in Africa, this study provides useful information for monitoring changes in parasite population structure in the face of ongoing and future control.
Meekums H, Hawash MB, Sparks AM, Oviedo Y, Sandoval C, Chico ME, Stothard JR, Cooper PJ, Nejsum P, Betson M (2015) A genetic analysis of Trichuris trichiura and Trichuris suis from Ecuador., Parasites & vectors 8
Since the nematodes Trichuris trichiura and T. suis are morphologically indistinguishable, genetic analysis is required to assess epidemiological cross-over between people and pigs. This study aimed to clarify the transmission biology of trichuriasis in Ecuador.Adult Trichuris worms were collected during a parasitological survey of 132 people and 46 pigs in Esmeraldas Province, Ecuador. Morphometric analysis of 49 pig worms and 64 human worms revealed significant variation. In discriminant analysis morphometric characteristics correctly classified male worms according to host species. In PCR-RFLP analysis of the ribosomal Internal Transcribed Spacer (ITS-2) and 18S DNA (59 pig worms and 82 human worms), nearly all Trichuris exhibited expected restriction patterns. However, two pig-derived worms showed a "heterozygous-type" ITS-2 pattern, with one also having a "heterozygous-type" 18S pattern. Phylogenetic analysis of the mitochondrial large ribosomal subunit partitioned worms by host species. Notably, some Ecuadorian T. suis clustered with porcine Trichuris from USA and Denmark and some with Chinese T. suis.This is the first study in Latin America to genetically analyse Trichuris parasites. Although T. trichiura does not appear to be zoonotic in Ecuador, there is evidence of genetic exchange between T. trichiura and T. suis warranting more detailed genetic sampling.
BACKGROUND: In 2012 the WHO formally recognised that infants and preschool children are at significant risk of schistosomiasis and qualify for treatment with praziquantel (PZQ). Targeted surveys determining both the performance and safety of this drug are now needed in endemic areas. We have formally assessed parasitological cure and putative side-effects in a prospective cohort of Schistosoma mansoni-infected children (aged 5 months-7 years old) in lakeshore settings of Uganda. METHODOLOGY/PRINCIPAL FINDINGS: From a total of 369 children found to be egg-patent for intestinal schistosomiasis, 305 were followed-up three to four weeks after PZQ treatment and infection status re-assessed. Separately, a previously tested side-effect questionnaire was employed before and 24 hours after PZQ treatment to assess incidence and amelioration of symptoms in young children and their mothers. While the overall observed parasitological cure was 56.4%, a significant difference was found between a sub-set of children who had a history of multiple PZQ treatments (between one and four in an 18 month period), where cure rate was 41.7%, and those who had never received treatment (cure rate was 77·6%). PZQ proved to be safe, with only mild reported side effects which cleared within a month after treatment. Prevalence of reported symptoms was significantly lower in children than in mothers, and fewer side-effects were reported upon subsequent rounds of PZQ treatment. CONCLUSION/SIGNIFICANCE: Our findings show that PZQ treatment of young children resulted in satisfactory cure rates, and marked reduction in egg-output, with only mild and transient reported side-effects. However, the cure rate is clearly lower in younger children and those with history of previous treatment. Cure rate, but not egg reduction rate, was also lower in children with heavier pre-intervention infection intensity. With chemotherapy now recommended as a long-term strategy for disease control in young children, research into optimising the periodicity of targeted treatment strategies is now crucial.
Marie H, Pratt SJ, Betson M, Epple H, Kittler JT, Meek L, Moss SJ, Troyanovsky S, Attwell D, Longmore GD, Braga VM (2003) The LIM protein Ajuba is recruited to cadherin-dependent cell junctions through an association with alpha-catenin., The Journal of biological chemistry 278 (2) pp. 1220-1228
Cell-cell adhesive events affect cell growth and fate decisions and provide spatial clues for cell polarity within tissues. The complete molecular determinants required for adhesive junction formation and their function are not completely understood. LIM domain-containing proteins have been shown to be present at cell-cell contact sites and are known to shuttle into the nucleus where they can affect cell fate and growth; however, their precise localization at cell-cell contacts, how they localize to these sites, and what their functions are at these sites is unknown. Here we show that, in primary keratinocytes, the LIM domain protein Ajuba is recruited to cadherin-dependent cell-cell adhesive complexes in a regulated manner. At cadherin adhesive complexes Ajuba interacts with alpha-catenin, and alpha-catenin is required for efficient recruitment of Ajuba to cell junctions. Ajuba also interacts directly with F-actin. Keratinocytes from Ajuba null mice exhibit abnormal cell-cell junction formation and/or stability and function. These data reveal Ajuba as a new component at cadherin-mediated cell-cell junctions and suggest that Ajuba may contribute to the bridging of the cadherin adhesive complexes to the actin cytoskeleton and as such contribute to the formation or strengthening of cadherin-mediated cell-cell adhesion.
Bustinduy A, King C, Scott J, Appleton S, Sousa-Figueiredo JC, Betson M, Stothard JR (2014) HIV and schistosomiasis co-infection in African children., The Lancet. Infectious diseases 14 (7) pp. 640-649
HIV/AIDS and schistosomiasis both cause a substantial disease burden in sub-Saharan Africa and the two diseases often overlap in their epidemiological characteristics. Although disease-specific control interventions are continuing, potential synergies in the control efforts for these two diseases have not been investigated. With a focus on children with schistosomiasis, we assess the risk for increased HIV transmission, HIV progression, and impaired response to drugs when given alongside HIV interventions. A new research agenda tailored to children is needed to better understand the interactions of these two diseases and the potential for combined responses.
Standley CJ, Mugisha L, Verweij JJ, Adriko M, Arinaitwe M, Rowell C, Atuhaire A, Betson M, Hobbs E, van Tulleken CR, Kane RA, van Lieshout L, Ajarova L, Kabatereine NB, Stothard JR (2011) Confirmed infection with intestinal schistosomiasis in semi-captive wild-born chimpanzees on Ngamba Island, Uganda., Vector borne and zoonotic diseases (Larchmont, N.Y.) 11 (2) pp. 169-176
BACKGROUND: Intestinal schistosomiasis, caused by Schistosoma mansoni, is endemic to Lake Victoria, with high prevalence of the disease observed in human lakeshore communities. However, nonhuman primates have recently been overlooked as potential hosts of the disease, despite known susceptibility. METHODS: Using a variety of stool, urine, and serological diagnostic methods, 39 semi-captive wild-born chimpanzees and 37 staff members at Ngamba Island Chimpanzee Sanctuary, Lake Victoria, Uganda, were examined for S. mansoni infection. Miracidia recovered from stool were DNA barcoded to investigate cross-over between humans and chimpanzees. The island was also surveyed for Biomphalaria intermediate host snails, which were examined for infection with S. mansoni. RESULTS: Chimpanzees were unequivocally shown to be infected with intestinal schistosomiasis with a seroprevalence in excess of 90%. Three egg-positive cases were detected, although the sensitivity of the diagnostic tests varied due to earlier prophylactic praziquantel treatment. Miracidia hatched from chimpanzee stool revealed three DNA haplotypes commonly found in humans living throughout Lake Victoria, including staff on Ngamba Island, as well as two novel haplotypes. At one site, a snail was observed shedding schistosome cercariae. CONCLUSIONS: The anthropozoonotic potential of intestinal schistosomiasis on Ngamba Island is greater than previously thought. Moreover, the ability of chimpanzees to void schistosome eggs capable of hatching into viable miracidia further suggests that these nonhuman primates may be capable of maintaining a local zoonotic transmission of schistosomiasis independently of humans. The implications for management of captive and wild primate populations at risk of exposure are discussed.
Betson M, Lozano E, Zhang J, Braga VM (2002) Rac activation upon cell-cell contact formation is dependent on signaling from the epidermal growth factor receptor., The Journal of biological chemistry 277 (40) pp. 36962-36969
Cadherins are transmembrane receptors that mediate cell-cell adhesion. They play an essential role in embryonic development and maintenance of tissue architecture. The Rho family small GTPases regulate actin cytoskeletal dynamics in different cell types. The function of two family members, Rho and Rac, is required for the stability of cadherins at cell-cell contacts. Consistent with the published data we have found that Rac is activated upon induction of intercellular adhesion in epithelial cells. This activation is dependent on functional cadherins (Nakagawa, M., Fukata, M., Yamaga, M., Itoh, N., and Kaibuchi, K. (2001) J. Cell Sci. 114, 1829-1838; Noren, N. K., Niessen, C. M., Gumbiner, B. M., and Burridge, K. (2001) J. Biol. Chem. 276, 3305-3308). Here we show for the first time that clustering of cadherins using antibody-coated beads is sufficient to promote Rac activation. In the presence of Latrunculin B, Rac can be partially activated by antibody-clustered cadherins. These results suggest that actin polymerization is not required for initial Rac activation. Contrary to what has been described before, phosphatidylinositol 3-kinases are not involved in Rac activation following cell-cell adhesion in keratinocytes. Interestingly, inhibition of epidermal growth factor receptor signaling efficiently blocks the increased Rac-GTP levels observed after contact formation. We conclude that cadherin-dependent adhesion can activate Rac via epidermal growth factor receptor signaling.
Betson M, Nejsum P, Stothard JR (2013) From the twig tips to the deeper branches: new insights into the evolutionary history and phylogeography of Ascaris, In: Holland C (eds.), Ascaris: the neglected parasite 10 Academic Press Elsevier
To shed light on the epidemiology of ascariasis in Ecuador and Zanzibar, 177 adult worms retrieved by chemo-expulsion from either people or pigs were collected, measured and subjected to polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the ribosomal internal transcribed spacer (ITS) region. Upon double digestion with RsaI and HaeIII, PCR-RFLP analysis revealed the presence of A. lumbricoides in people and A. suum in pigs in Ecuador. In contrast, while there are no pigs on Zanzibar, of the 56 worms obtained from people, one was genotyped as A. suum. No additional genetic variation was detected upon further PCR-RFLP analysis with several other restriction enzymes. Upon measurement, worm mass and length differed by location and by species, A. suum being lighter and longer. While there is no evidence to suggest zoonotic transmission in Ecuador, an enduring historical signature of previous zoonotic transmission remains on Zanzibar.
Ascaris lumbricoides and A. suum are two parasitic nematodes infecting humans and pigs, respectively. There has been considerable debate as to whether Ascaris in the two hosts should be considered a single or two separate species. Previous studies identified at least three major clusters (A, B and C) of human and pig Ascaris based on partial cox1 sequences. In the present study, we selected major haplotypes from these different clusters to characterize their whole mitochondrial genomes for phylogenetic analysis. We also undertook coalescent simulations to investigate the evolutionary history of the different Ascaris haplotypes. The topology of the phylogenetic tree based on complete mitochondrial genomic sequences was found to be similar to partial cox1 sequencing, but the support at internal nodes was higher in the former. Coalescent simulations suggested the presence of at least two divergence events: the first one occurring early in the Neolithic period which resulted in a differentiated population of Ascaris in pigs (cluster C), the second occurring more recently (~900 generations ago), resulting in clusters A and B which might have been spread worldwide by human activities.
As part of a longitudinal cohort investigation of intestinal schistosomiasis and malaria in Ugandan children and their mothers on the shorelines of Lakes Victoria and Albert, we documented risk factors and morbidity associated with nonfalciparum Plasmodium infections and the longitudinal dynamics of Plasmodium species in children. Host age, household location, and Plasmodium falciparum infection were strongly associated with nonfalciparum Plasmodium infections, and Plasmodium malariae infection was associated with splenomegaly. Despite regular artemisinin combination therapy treatment, there was a 3-fold rise in P. malariae prevalence, which was not accountable for by increasing age of the child. Worryingly, our findings reveal the consistent emergence of nonfalciparum infections in children, highlighting the complex dynamics underlying multispecies infections here. Given the growing body of evidence that nonfalciparum malaria infections cause significant morbidity, we encourage better surveillance for nonfalciparum Plasmodium infections, particularly in children, with more sensitive DNA detection methods and improved field-based diagnostics.
Human trichuriasis is a neglected tropical disease
which affects hundreds of millions of people worldwide and is
particularly prevalent among children living in areas where
sanitation is poor. This review examines the current knowledge
on the taxonomy, genetics and phylogeography of human
Trichuris and its relationship to whipworm parasites in
other host species. The evidence for zoonotic transmission of
Trichuris and the emergence of anthelmintic resistance is
assessed. In addition, the implications of the recent publication
of the genomes and transcriptomes of multiple Trichuris
species are discussed. Finally, priorities for future research in
Trichuris genetics are proposed.
Surrey, a county in southern England, is a hot spot for angiostrongylosis in domestic dogs but there have been no investigations into the intermediate hosts of Angiostrongylus vasorum in this area. This study aimed to determine the prevalence of A. vasorum in gastropods in Guildford, the most populous town in Surrey, and to ascertain which gastropod species can act as intermediate hosts for A. vasorum. Gastropods (n=97) were sampled in six locations, representing urban, suburban and rural environments and identified to species based on morphological features. A PCR assay was used to detect A. vasorum DNA in gastropod tissue and the species of infected specimens was confirmed by sequencing of mitochondrial genes. 4.1% (4/97) of sampled gastropods and 9.1% (4/44) of sampled slugs were A. vasorum positive. Infected gastropod species were Arion rufus (n=3) and Deroceras invadens (n=1), the first description of the latter species as a potential intermediate host for A. vasorum. Two infected slugs were sampled in urban environments and two in suburban environments. The results demonstrate that there is a risk of transmission of A. vasorum to domestic dogs from the gastropod population in urban and suburban areas of Guildford.
Culicoides biting midges are biological vectors of arthropod-borne viruses (arboviruses) of international importance. The emergence of bluetongue virus (BTV) and Schmallenberg virus (SBV) in Europe has highlighted the vulnerability of the UK to incursions. Persistence of Culicoides-borne viruses through adverse conditions in the cold winter months (?overwintering?) contributes to their economic and welfare impact and is a major source of uncertainty in policy response. Understanding the role of temperature in adult Culicoides activity, particularly in colder months, will inform policy on the likely risk of transmission. Reducing exposure to Culicoides biting is an important factor in controlling the spread of Culicoides-borne viruses following an incursion and immunological responses to the Culicoides bite may provide a novel approach to measuring the exposure of susceptible hosts to Culicoides.
This thesis investigates adult Culicoides seasonal activity and biting in the UK using both field and laboratory-based studies, focusing on the impact of winter. Culicoides activity, examined directly using ultraviolet-baited light-suction traps, was limited to days when maximum temperatures were >7°C in a field study. An experimental system that tested movement of adult Culicoides within a controlled environment further demonstrated the effect of temperature, with the minimum temperature for flight activity varying according to collection location, time of year of collection and species. Furthermore, immunoassays were developed to detect the presence of antibodies against Culicoides salivary proteins in host sera and to indirectly infer Culicoides blood-feeding activity. This work demonstrated that horses, cattle and sheep in the UK produce detectable levels of antibodies recognising Culicoides salivary protein antigens for the first time, as a result of natural biting exposure in the field environment.
These findings have direct implications in future surveillance programs and policy decision making, particularly when defining vector-free periods of adult activity.