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Systematic Review of Wolbachia Symbiont Detection in Mosquitoes: An Entangled Topic about Methodological Power and True Symbiosis.

ABSTRACT: Wolbachia is an endosymbiotic bacterium that naturally infects several arthropods and nematode species. Wolbachia gained particular attention due to its impact on their host fitness and the capacity of specific Wolbachia strains in reducing pathogen vector and agricultural pest populations and pathogens transmission. Despite the success of mosquito/pathogen control programs using Wolbachia-infected mosquito release, little is known about the abundance and distribution of Wolbachia in most mosquito species, a crucial knowledge for planning and deployment of mosquito control programs and that can further improve our basic biology understanding of Wolbachia and host relationships. In this systematic review, Wolbachia was detected in only 30% of the mosquito species investigated. Fourteen percent of the species were considered positive by some studies and negative by others in different geographical regions, suggesting a variable infection rate and/or limitations of the Wolbachia detection methods employed. Eighty-three percent of the studies screened Wolbachia with only one technique. Our findings highlight that the assessment of Wolbachia using a single approach limited the inference of true Wolbachia infection in most of the studied species and that researchers should carefully choose complementary methodologies and consider different Wolbachia-mosquito population dynamics that may be a source of bias to ascertain the correct infectious status of the host species.

SUBMITTER: Inacio da Silva LM

PROVIDER: S-EPMC7825316 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature

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Systematic Review of Wolbachia Symbiont Detection in Mosquitoes: An Entangled Topic about Methodological Power and True Symbiosis.

Inácio da Silva Luísa Maria LM Dezordi Filipe Zimmer FZ Paiva Marcelo Henrique Santos MHS Wallau Gabriel Luz GL

Pathogens (Basel, Switzerland) 20210106 1

Wolbachia is an endosymbiotic bacterium that naturally infects several arthropods and nematode species. Wolbachia gained particular attention due to its impact on their host fitness and the capacity of specific Wolbachia strains in reducing pathogen vector and agricultural pest populations and pathogens transmission. Despite the success of mosquito/pathogen control programs using Wolbachia-infected mosquito release, little is known about the abundance and distribution ...[more]

PMID: 33419044

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Project description:BackgroundOnchocerciasis (river blindness) and lymphatic filariasis (elephantiasis) are two human neglected tropical diseases that cause major disabilities. Mass administration of drugs targeting the microfilarial stage has reduced transmission and eliminated these diseases in several countries but a macrofilaricidal drug that kills or sterilizes the adult worms is critically needed to eradicate the diseases. The causative agents of onchocerciasis and lymphatic filariasis are filarial worms that harbor the endosymbiotic bacterium Wolbachia. Because filarial worms depend on Wolbachia for reproduction and survival, drugs targeting Wolbachia hold great promise as a means to eliminate these diseases.MethodsTo better understand the relationship between Wolbachia and its worm host, adult Brugia pahangi were exposed to varying concentrations of doxycycline, minocycline, tetracycline and rifampicin in vitro and assessed for Wolbachia numbers and worm motility. Worm motility was monitored using the Worminator system, and Wolbachia titers were assessed by qPCR of the single copy gene wsp from Wolbachia and gst from Brugia to calculate IC50s and in time course experiments. Confocal microscopy was also used to quantify Wolbachia located at the distal tip region of worm ovaries to assess the effects of antibiotic treatment in this region of the worm where Wolbachia are transmitted vertically to the microfilarial stage.ResultsWorms treated with higher concentrations of antibiotics had higher Wolbachia titers, i.e. as antibiotic concentrations increased there was a corresponding increase in Wolbachia titers. As the concentration of antibiotic increased, worms stopped moving and never recovered despite maintaining Wolbachia titers comparable to controls. Thus, worms were rendered moribund by the higher concentrations of antibiotics but Wolbachia persisted suggesting that these antibiotics may act directly on the worms at high concentration. Surprisingly, in contrast to these results, antibiotics given at low concentrations reduced Wolbachia titers.ConclusionWolbachia in B. pahangi display a counterintuitive dose response known as the "Eagle effect." This effect in Wolbachia suggests a common underlying mechanism that allows diverse bacterial and fungal species to persist despite exposure to high concentrations of antimicrobial compounds. To our knowledge this is the first report of this phenomenon occurring in an intracellular endosymbiont, Wolbachia, in its filarial host.

Project description:Manipulation of natural mosquito populations using the endosymbiotic bacteria Wolbachia is being investigated as a novel strategy to reduce the burden of mosquito-borne viruses. To evaluate the efficacy of these interventions, it will be critical to determine Wolbachia infection frequencies in Aedes aegypti mosquito populations. However, current diagnostic tools are not well-suited to fit this need. Morphological methods cannot identify Wolbachia, immunoassays often suffer from low sensitivity and poor throughput, while PCR and spectroscopy require complex instruments and technical expertise, which restrict their use to centralized laboratories. To address this unmet need, we have used loop-mediated isothermal amplification (LAMP) and oligonucleotide strand displacement (OSD) probes to create a one-pot sample-to-answer nucleic acid diagnostic platform for vector and symbiont surveillance. LAMP-OSD assays can directly amplify target nucleic acids from macerated mosquitoes without requiring nucleic acid purification and yield specific single endpoint yes/no fluorescence signals that are observable to eye or by cellphone camera. We demonstrate cellphone-imaged LAMP-OSD tests for two targets, the Aedes aegypti cytochrome oxidase I (coi) gene and the Wolbachia surface protein (wsp) gene, and show a limit of detection of 4 and 40 target DNA copies, respectively. In a blinded test of 90 field-caught mosquitoes, the coi LAMP-OSD assay demonstrated 98% specificity and 97% sensitivity in identifying Ae. aegypti mosquitoes even after 3 weeks of storage without desiccant at 37°C. Similarly, the wsp LAMP-OSD assay readily identified the wAlbB Wolbachia strain in field-collected Aedes albopictus mosquitoes without generating any false positive signals. Modest technology requirements, minimal execution steps, simple binary readout, and robust accuracy make the LAMP-OSD-to-cellphone assay platform well suited for field vector surveillance in austere or resource-limited conditions.

Dataset Information

Systematic Review of Wolbachia Symbiont Detection in Mosquitoes: An Entangled Topic about Methodological Power and True Symbiosis.

Publications

Systematic Review of <i>Wolbachia</i> Symbiont Detection in Mosquitoes: An Entangled Topic about Methodological Power and True Symbiosis.

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