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Discrimination between Onchocerca volvulus and O. ochengi filarial larvae in Simulium damnosum (s.l.) and their distribution throughout central Ghana using a versatile high-resolution speciation assay

Stephen R. Doyle, Samuel Armoo, Alfons Renz, Mark J Taylor, Mike Yaw Osei-Atweneboana, Warwick N Grant
doi: https://doi.org/10.1101/046987
Stephen R. Doyle
1Department of Animal, Plant and Soil Sciences, La Trobe University, Bundoora, 3086, Australia
5current address: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridgeshire, CB10 1SA, United Kingdom
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  • For correspondence: stephenrdoyle1@gmail.com
Samuel Armoo
1Department of Animal, Plant and Soil Sciences, La Trobe University, Bundoora, 3086, Australia
2Council for Scientific and industrial Research - Water Research Institute, Accra, Ghana
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Alfons Renz
3Institute of Evolution and Ecology, Department of Comparative Zoology, University of Tübingen, Auf der Morgenstelle 28, 74074, Tübingen, Germany
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Mark J Taylor
4Department of Parasitology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom
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Mike Yaw Osei-Atweneboana
2Council for Scientific and industrial Research - Water Research Institute, Accra, Ghana
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Warwick N Grant
1Department of Animal, Plant and Soil Sciences, La Trobe University, Bundoora, 3086, Australia
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Abstract

Background Genetic surveillance of the human filarial parasite, Onchocerca volvulus, from onchocerciasis endemic regions will ideally focus on genotyping individual infective larval stages collected from their intermediate host, Simuliid blackflies. However, blackflies also transmit other Onchocerca species, including the cattle parasite O. ochengi, which are difficult to distinguish from the human parasite based on morphological characteristics alone. This study describes a versatile approach to discriminate between O. volvulus and O. ochengi that is demonstrated using parasite infective larvae dissected from blackflies.

Results A speciation assay was designed based on genetic differentiation between O. volvulus and O. ochengi mitochondrial genome sequences that can be performed in highthroughput high-resolution melt (HRM)- or lower throughput conventional restriction fragment length polymorphism (RFLP) analyses. This assay was validated on 185 Onchocerca larvae dissected from blackflies captured from 14 communities in Ghana throughout 2011-13. The frequency of O. ochengi was approximately 67 % of all larvae analysed, which is significantly higher than previously reported in this region. Furthermore, the species distribution was not uniform throughout the study region, with 25 %, 47 % and 93 % of O. volvulus being found in the western-most (Black Volta, Tain and Tombe), the central (Pru) and eastern-most (Daka) river basins, respectively.

Conclusions This tool provides a simple and cost-effective approach to determine the identity and distribution of two Onchocerca species, and will be valuable for future genetic studies that focus on parasites collected from blackflies. The results presented highlight the need to discriminate Onchocerca species in transmission studies, as the frequency of each species varied significantly between the communities studied.

Footnotes

  • ↵† Equal contributions

  • Emails: Stephen R Doyle: stephenrdoyle1{at}gmail.com, Samuel Armoo: sarmoo{at}students.latrobe.edu.au, Alfons Renz: Alfons.Renz{at}uni-tuebingen.de, Mark J Taylor: mark.taylor{at}lstmed.ac.uk, Mike Yaw Osei-Atweneboana: oseiatweneboana{at}yahoo.co.uk, Warwick N Grant: w.grant{at}latrobe.edu.au

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted October 01, 2016.
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Discrimination between Onchocerca volvulus and O. ochengi filarial larvae in Simulium damnosum (s.l.) and their distribution throughout central Ghana using a versatile high-resolution speciation assay
Stephen R. Doyle, Samuel Armoo, Alfons Renz, Mark J Taylor, Mike Yaw Osei-Atweneboana, Warwick N Grant
bioRxiv 046987; doi: https://doi.org/10.1101/046987
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Discrimination between Onchocerca volvulus and O. ochengi filarial larvae in Simulium damnosum (s.l.) and their distribution throughout central Ghana using a versatile high-resolution speciation assay
Stephen R. Doyle, Samuel Armoo, Alfons Renz, Mark J Taylor, Mike Yaw Osei-Atweneboana, Warwick N Grant
bioRxiv 046987; doi: https://doi.org/10.1101/046987

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