Characterization of Ascaris from Ecuador and Zanzibar

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

Use of synchrotron tomographic techniques in the assessment of diffusion parameters for solute transport in groundwater flow

This technical note describes the use of time-resolved synchrotron radiation tomographic energy dispersive diffraction imaging (TEDDI) and tomographic X-ray fluorescence (TXRF) for examining ion diffusion in porous media. The technique is capable of tracking the diffusion of several ion species simultaneously. This is illustrated by results which compare the movement of Cs+, Ba2+ and La3+ ions from solution into a typical sample of English chalk. The results exhibited somewhat anomalous (non-Fickian) behaviour and revealed heterogeneities (in 1D) on the scale of a few millimetres

Actin at cell-cell junctions is composed of two dynamic and functional populations

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

Environmental Epidemiology of Intestinal Schistosomiasis in Uganda: Population Dynamics of Biomphalaria (Gastropoda: Planorbidae) in Lake Albert and Lake Victoria with Observations on Natural Infections with Digenetic Trematodes

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 < 0.001) in both lakes and negative relationship with conductivity (P = 0.04) in Lake Albert. Of the Biomphalaria collected in Lake Albert (N = 6,183), 8.9% were infected with digenetic trematodes of which 15.8% were shedding S. mansoni cercariae and 84.2% with nonhuman infective cercariae. In Lake Victoria, 2.1% of collected Biomphalaria (N = 13,172) were infected with digenetic trematodes with 13.9% shedding S. mansoni cercariae, 85.7% shedding nonhuman infective cercariae, and 0.4% of infected snails shedding both types of cercariae. Upon morphological identification, species of Biomphalaria infected included B. sudanica, B. pfeifferi, and B. stanleyi in Lake Albert and B. sudanica, B. pfeifferi, and B. choanomphala in Lake Victoria. The study found the physicochemical factors that influenced Biomphalaria population and infections. The number and extent of snails shedding S. mansoni cercariae illustrate the high risk of transmission within these lake settings. For better control of this disease, greater effort should be placed on reducing environmental contamination by improvement of local water sanitation and hygiene

Molecular Epidemiology of Ascariasis: A Global Perspective on the Transmission Dynamics of Ascaris in People and Pigs

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 program

Whipworms in humans and pigs: origins and demography

© 2016 Hawash et al. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. The attached file is the published version of the article.NHM Repositor

Validation of deep amplicon sequencing of Dicrocoelium in small ruminants from Northern regions of Pakistan.

Dicrocoelium lancet flukes cause significant production loss in ruminant livestock. Although co-infection with multiple Dicrocoelium species within a host is common, techniques for studying the composition of these complex parasite communities are lacking. The pathogenicity, epidemiology, and therapeutic susceptibility of different helminth species vary, and little is known about the interactions that take place between co-infecting species and their hosts. Here, we describe the first applicationof metabarcoding deep amplicon sequencingmethod to studythe Dicrocoelium species in sheep and goats. First, rDNA ITS-2 sequences of four Dicrocoelium species (Dicrocoelium dendriticum, Dicrocoelium hospes, Dicrocoelium orientalis, and Dicrocoelium chinensis) were extracted from the NCBI public database. Phylogenetic analysis revealed separate clades of Dicrocoelium species; hence, molecular differentiation between each species is possible in co-infections. Second, 202 flukes belonging to seventeen host populations (morphologically verified as belonging to the Dicrocoelium genus) were evaluated to determine the deep amplicon sequencing read thresholdof an individual fluke for each of the four species. The accuracy of the method in proportional quantification of samples collected from single hosts was further assessed. Overall, 198 (98.01%) flukes were confirmed as D. dendriticum and 1.98% produced no reads. The comparison of genetic distances between rDNA ITS-2 revealed 86% to 98% identity between the Dicrocoelium species. Phylogenetic analysis demonstrated a distinct clustering of species, apart from D. orientalis and D. chinensis, which sit very close to each other in a singlelarge clade whereas D. hospes and D. dendriticum are separated into their own clade. In conclusion each sample was identified as D. dendriticum based on the proportion of MiSeq reads and validated the presence of this group of parasites in the Gilgit Baltistan and Khyber Pakhtunkhwa provinces of Pakistan. The metabarcoding deep amplicon sequencing technology and bioinformatics pathway have several potential applications, including speciesinteractions during co-infections, identifying the host and geographical distribution of Dicrocoelium in livestock, drug therapy response evaluation and understanding of the emergence and spread of drug resistance

Fecal occult blood and fecal calprotectin as point-of-care markers of intestinal morbidity in Ugandan children with Schistosoma mansoni infection.

BACKGROUND: Calprotectin is a calcium-binding cytoplasmic protein found in neutrophils and increasingly used as a marker of bowel inflammation. Fecal occult blood (FOB) is also a dependable indicator of bowel morbidity. The objective of our study was to determine the applicability of these tests as surrogate markers of Schistosoma mansoni intestinal morbidity before and after treatment with praziquantel (PZQ). METHODS: 216 children (ages 3-9 years old) from Buliisa District in Lake Albert, Uganda were examined and treated with PZQ at baseline in October 2012 with 211 of them re-examined 24 days later for S. mansoni and other soil transmitted helminths (STH). POC calprotectin and FOB assays were performed at both time points on a subset of children. Associations between the test results and infection were analysed by logistic regression. RESULTS: Fecal calprotectin concentrations of 150-300 µg/g were associated with S. mansoni egg patent infection both at baseline and follow up (OR: 12.5 P = 0.05; OR: 6.8 P = 0.02). FOB had a very strong association with baseline anemia (OR: 9.2 P = 0.03) and medium and high egg intensity schistosomiasis at follow up (OR: 6.6 P = 0.03; OR: 51.3 P = 0.003). Both tests were strongly associated with heavy intensity S. mansoni infections. There was a significant decrease in FOB and calprotectin test positivity after PZQ treatment in those children who had egg patent schistosomiasis at baseline. CONCLUSIONS: Both FOB and calprotectin rapid assays were found to correlate positively and strongly with egg patent S. mansoni infection with a positive ameloriation response after PZQ treatment indicative of short term reversion of morbidity. Both tests were appropriate for use in the field with excellent operational performance and reliability. Due to its lower-cost which makes its scale-up of use affordable, FOB could be immediately adopted as a monitoring tool for PC campaigns for efficacy evaluation before and after treatment

Status of insecticide susceptibility in Anopheles gambiae s.l. from malaria surveillance sites in The Gambia

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 <80% but 88% for permethrin and 92% for deltamethrin. CONCLUSION: This current survey of insecticide resistance in Anopheles provides baseline information for monitoring resistance in The Gambia and highlights the need for routine resistance surveillance as an integral part of the proposed nation wide IRS intervention using DDT