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Gilleard, J.

Publications and source records attributed to Gilleard, J..

3 recordsLinked to original sources

Development, field-testing and optimization of tools to quantify soil-transmitted helminths in fecal sludge from school pit latrines in Ethiopia

BackgroundSurveys to monitor large-scale deworming programs against soil-transmitted helminthiases (STH) involve examination of stool samples from schoolchildren. These surveys are resource demanding and impact school activities. A potentially cost-saving alternative that does not involve children is to process fecal sludge samples from school pit latrines. To provide a proof-of-principle of latrine-based monitoring of STH programs, we optimized tools to collect fecal sludge and to quantify STH eggs in the samples. MethodsFirst, we designed, developed and field-tested three locally made fecal sludge sampling prototypes. Second, we developed a modified egg-counting method and conducted spiking experiments to explore its analytical performance. Third, we estimated the variation in egg counts in fecal sludge samples collected from six primary schools in Ethiopia at different pit latrine depths used by boys and girls and by repeatedly examining samples. Finally, field data were used to inform an egg count simulation model to quantify this variation in egg counts and to determine the sampling and analysis strategies that resulted in surveys as precise as a stool-based survey. ResultsThe modified fecal sludge sampling prototypes were generally successful, except for a few pit latrines with dried/solid sludge types and insufficient sludge volume. The egg-counting method had moderately high analytical sensitivity that varied across the consistency of the samples. The variation in egg counts was mainly explained by differences between squat holes followed by repeated fecal sludge sample processing. Latrine-based surveys were as precise as stool-based surveys only for Ascaris and when the intensity of infections was low. ConclusionsWe developed a sampling and diagnostic strategy that we will use in a follow-up study. This study will be conducted in Jimma Zone across 25 schools (52 children per school) and will compare the mean fecal sludge egg counts at school level with the STH prevalence in children. Author summaryProgress of large-scale deworming programs are currently monitored through screening individual stool samples of schoolchildren. A latrine-based program monitoring, without active participation of schoolchildren and interruption of routine school activities, is a potentially cost-effective alternative. As a proof-of-concept, we developed a method to quantify worm eggs in fecal sludge samples, conducted spiking experiments to determine its analytical performance and applied the method across schools in Ethiopia to explore variation in egg counts (e.g., squat holes, depth of sample collection, and repeated analyses). Based on our findings, we then determined the sample collection and analysis strategy that results a latrine-based survey as precise as a survey based on screening individual stool samples. The analytical performance was moderately high but varied across the consistency of the samples. The variation in egg counts was mainly driven by variation between squat holes on the same pit latrine, and thus it is better to sample more squat holes (at least 3) than repeatedly processing the same fecal sludge sample (one examination is sufficient). We will now apply this sample collection and analysis across 25 schools (52 children per school) in Ethiopia and compare it with a survey based on screening individual stool samples.

microbiology↗

Multiple Drug Resistance in the canine hookworm Ancylostoma caninum: an Emerging Threat

In the past few years, diagnoses by veterinarians of recurrent canine hookworm infections have dramatically increased, suggesting that anthelmintic resistance (AR) may have evolved in the parasite Ancylostoma caninum. To investigate this, we established three \"suspected-resistant\" and two susceptible A. caninum isolates in research dogs for further study. The egg hatch assay (EHA) and the larval development assay (LDA) were used for detecting resistance to benzimidazoles, and macrocyclic lactones, respectively. Resistance ratios ranged from 6.0 to >100 and 5.5-69.8 for the EHA and LDA, respectively. Following treatments with fenbendazole, pyrantel and milbemycin oxime, reduction in faecal egg counts ranged from 64-86%, 0-72% and 58-92%, respectively. Deep amplicon sequencing of the isotype-1 {beta} tubulin gene identified a high frequency of resistance-associated single nucleotide polymorphisms at codon 167 in the resistant isolates and clinical cases.. These data conclusively demonstrate multiple anthelmintic resistance in A. caninum, and provide pivotal evidence that this is an emerging problem in the United States. Consequently, these findings should provide some concern to the global health community, as the scale-up of mass drug administration for soil-transmitted helminths (STH) is now placing similar selection pressures for benzimidazole resistance in human hookworms.

zoology↗

Evidence for the F200Y (TAC) mutation conferring benzimidazole resistance in a southern USA cattle population of Haemonchus placei spreading from a single emergence

The benzimidazoles are one of the most important broad-spectrum anthelmintic drug classes for the control of parasitic nematodes of domestic animals and humans. They have been widely used in the livestock sector, particularly in small ruminants for over 40 years. This has resulted in the development and wide spread of resistance in small ruminant gastrointestinal nematode parasite species, including Haemonchus contortus. Recently, resistance to benzimidazole drugs has been reported in Haemonchus placei, but there is relatively little information on its prevalence. It is important to develop a molecular tools to identify resistance mutations in H. placei early in their development in order to understand the emergence and spread. Our previous study demonstrated the F200Y (TAC) mutation at their early stage in 6/9 H. placei populations derived from southern USA, albeit at low frequencies between 2 and 10%. The present study analysis the phylogenetics of the isotype-1 {beta}-tubulin locus to suggest that F200Y (TAC) mutation has been spread from a single emergence in H. placei; likely by the anthroprogenic movement of ruminant livestock in southern USA. Population genetic data of H. placei using a panel of microsatellite markers revealed little genetic sub-structure, consistent with a high level of gene flow in this region. Overall, these results provide clear genetic evidence for the spread of F200Y (TAC) benzimidazoles resistance mutation to multiple different locations from a single emergence in H. placei.

molecular biology↗