Biology subjects

Hackett, T. D.

Publications and source records attributed to Hackett, T. D..

2 recordsLinked to original sources

bioRxiv2026

A novel qPCR assay to detect the presence of Anopheles gambiae complex mosquitoes

BackgroundAnopheles mosquitoes vector pathogens responsible for more than 600,000 human deaths annually. Ecological studies of these insects are important to guide effective vector-control campaigns and to understand their broader ecological consequences. Molecular ecology methods, particularly qPCR, provide a valuable tool in such studies. By detecting trace DNA of a taxon of interest within mixed or environmental samples, qPCR can facilitate identification of prey taxa of interest in the diets of consumers. However, no protocol for the detection of An. gambiae complex mosquitoes in dietary samples has been available. MethodsWe introduce a new set of qPCR primers (Agam_CO1_F1 and Agam_CO1_R1) and a probe-based assay for detection of Anopheles gambiae-complex mosquitoes, even with short reads common in dietary and environmental samples. The primers were tested in vitro for their specificity and sensitivity, and in silico using Primer-BLAST to assess potential off-target amplification. ResultsThe qPCR primers amplified An. gambiae DNA even at low starting concentrations (5 copies {micro}l-1). The primers did not amplify any non-target DNA in either the in vitro or in silico tests, but consistently amplified An. gambiae complex DNA. The primers can therefore provide reliable tests for the presence or absence of An. gambiae complex in dietary or eDNA samples. ConclusionsThe new qPCR primers should allow advances in research into mosquito ecology by allowing detection of even trace amounts of An. gambiae DNA in dietary and environmental samples. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=72 SRC="FIGDIR/small/707393v1_ufig1.gif" ALT="Figure 1"> View larger version (11K): org.highwire.dtl.DTLVardef@1fcce3corg.highwire.dtl.DTLVardef@47e5f5org.highwire.dtl.DTLVardef@4a7063org.highwire.dtl.DTLVardef@1188d60_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

bioRxiv2025

Characterising a species-rich and understudied tropical insect fauna using DNA barcoding

BackgroundWest Africa has high biodiversity that is relatively understudied, especially for insects. Studies of West African arthropod diversity can therefore help address important questions regarding conservation, ecosystem services, and insecticide use and other species-control interventions in agriculture and disease management. We intensively sampled arthropods in Ghana using complementary trapping methods, generated DNA barcodes, and classified sequences by Barcode Index Numbers (BINs, a species proxy). Using this dataset, we investigate assemblage composition, temporal activity patterns, and the state of regional biodiversity sampling. ResultsSequencing DNA from 95,996 individuals captured using Malaise, yellow pan, pitfall, Heath and Centre for Disease Control (CDC) traps, we identified 10,120 unique BINs. The rate of species accumulation did not approach an asymptote for any taxonomic group or trap type, indicating high biodiversity. The different trap types sampled different subsets of the local community, with greatest similarity between yellow pan and pitfall traps. More insects and species (BINs) were trapped during the day than at night. Our dataset shared more BINs in the Barcode of Life Database with South Africa than with any other country, although this predominantly reflects the limited sampling and DNA sequencing campaigns in Africa. ConclusionsThis study more than doubles the published BINs for West Africa, offering insights into the biodiversity of an ecologically important but understudied taxon and region. Using multiple trap types allowed a more complete assessment of the local arthropod assemblage. The public release of these data will support and stimulate further taxonomic and ecological work in the region.

ecology↗