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. 2023 Feb 15;13:2729. doi: 10.1038/s41598-023-29657-y

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Design of experiments to investigate the impact of different environmental conditions on An. stephensi and Ae. aegypti eDNA detection. Numbers of second instar larvae are denoted on each artificial breeding site. Experiment 1 investigated eDNA detection of different densities of An. stephensi and Ae. aegypti in 50 ml artificial breeding sites. Experiment 2 investigated eDNA detection of different densities of An. stephensi and Ae. aegypti in 1L artificial breeding sites, including different ratios of An. stephensi:Ae. aegypti co-habiting the same breeding site. Experiment 3 investigated the rate of eDNA degradation. An. stephensi pupae were left in each breeding site for 24 h, prior to manual removal of all emerged adults, remaining pupae, and pupal skins; eDNA detection was performed for the following 14 days. Figure created with BioRender.com.