Abstract
Emerging resistance in malaria vectors and parasites has accelerated the development of novel control strategies such as the Sterile Insect Technique and Wolbachia-based approaches, which require large-scale mosquito rearing. These methods traditionally rely on vertebrate Blood, raising ethical and logistical concerns. This study evaluated the effects of a BLOODless™ on wing size, longevity, and Plasmodium falciparum susceptibility in Anopheles coluzzii and An. gambiae from Burkina Faso, and the Kisumu An. gambiae strain. Blood and Bloodless-reared mosquitoes were compared across seven generations. Wing size was unaffected by diet in the 3rd and 7th generations, though sex impacted size in the 3rd generation only. A diet-species interaction was detected in the 7th generation. Plasmodium falciparum infection rates were similar in the 3rd generation, but by the 6th, Blood-fed mosquitoes showed higher oocyst prevalence, though intensity remained unchanged. Infection outcomes were species-dependent only at this later stage. Longevity, initially comparable across groups, increased significantly in Bloodless-fed mosquitoes by the 7th generation. Species had no effect on survival. A Blood-free diet enables sustained mosquito rearing while preserving traits essential for malaria research, providing an ethical, effective alternative to vertebrate Blood in vector control and experimental programs.
Data availability
The data generated in this study are available as Additional file 1.
References
WHO. World malaria report. (2024).
Hemingway, J. et al. Tools and strategies for malaria control and elimination: what do we need to achieve a grand convergence in malaria? PLoS Biol. https://doi.org/10.1371/journal.pbio.1002380 (2016).
Ranson, H. & Lissenden, N. Insecticide resistance in African Anopheles mosquitoes: A worsening situation that needs urgent action to maintain malaria control. Trends Parasitol. https://doi.org/10.1016/j.pt.2015.11.010 (2016).
Ranson, H. Current and future prospects for preventing malaria transmission via the use of insecticides. Cold Spring Harbor Perspect. Med. https://doi.org/10.1101/cshperspect.a026823 (2017).
Bhatt, S. et al. The effect of malaria control on Plasmodium falciparum in Africa between 2000 and 2015. Nature (2015) https://doi.org/10.1038/nature15535
Huijben, S. & Paaijmans, K. P. Putting evolution in elimination: winning our ongoing battle with evolving malaria mosquitoes and parasites. Evol. Appl. https://doi.org/10.1111/eva.12530 (2018).
Sternberg, E. D. & Thomas, M. B. Insights from agriculture for the management of insecticide resistance in disease vectors. Evol. Appl. https://doi.org/10.1111/eva.12501 (2018).
WHO. World Malaria Report. (2022).
WHO. World malaria report. (2023).
Datoo, M. S. et al. Safety and efficacy of malaria vaccine candidate R21/Matrix-M in African children: a multicentre, double-blind, randomised, phase 3 trial. Lancet https://doi.org/10.1016/S0140-6736(23)02511-4 (2024).
Bouyer, J. Current status of the sterile insect technique for the suppression of mosquito populations on a global scale. Infect. Dis. Poverty. 13, 68 (2024).
Hammond, A. et al. A CRISPR-Cas9 gene drive system targeting female reproduction in the malaria mosquito vector Anopheles Gambiae. Nat. Biotechnol. https://doi.org/10.1038/nbt.3439 (2016).
Moreira, L. A. et al. A wolbachia symbiont in Aedes aegypti limits infection with Dengue, Chikungunya, and plasmodium. Cell 139, 1268–1278 (2009).
Yen, P. S. & Failloux, A. B. A review: Wolbachia-based population replacement for mosquito control shares common points with genetically modified control approaches. Pathogens https://doi.org/10.3390/pathogens9050404 (2020).
Sinden, R. E. Developing transmission-blocking strategies for malaria control. PLoS Pathog. 13, 1–12 (2017).
Geoffrey, M., Attardo, I. A. & Hansen, A. S. R. Nutritional regulation of vitellogenesis in mosquitoes: implications for anautogeny. Insect Biochem. Mol. Biol 35, (2005).
Marques, J. et al. Long-term blood-free rearing of Anopheles mosquitoes with no effect on fitness, Plasmodium infectivity nor microbiota composition. Sci. Rep. 14, 1–12 (2024).
Gonzales, K. K. & Hansen, I. A. Artificial diets for mosquitoes. Int. J. Environ. Res. Public Health. https://doi.org/10.3390/ijerph13121267 (2016).
Phasomkusolsil, S. et al. Maintenance of mosquito vectors: effects of blood source on feeding, survival, fecundity, and egg hatching rates. J. Vector Ecol. https://doi.org/10.1111/j.1948-7134.2013.12006.x (2013).
Marques, J. et al. Fresh-blood-free diet for rearing malaria mosquito vectors. Sci. Rep. https://doi.org/10.1038/s41598-018-35886-3 (2018).
Marques, J., Cardoso, J. C. R., Félix, R. C., Power, D. M. & Silveira, H. A Blood-Free diet to Rear Anopheline mosquitoes. J. Vis. Exp. https://doi.org/10.3791/60144-v (2020).
Mosi, F. A. et al. Effects of a blood-free mosquito diet on fitness and gonotrophic cycle parameters of laboratory reared Anopheles Gambiae sensu stricto. Parasites Vectors. 17, 1–8 (2024).
Ilboudo, H. et al. Insecticide resistance in Anopheles Gambiae in villages around the soum agropolis in the Center – West region of Burkina Faso. (2025).
Amara, M. F., Namountougou, M. & Konaté, H. Insecticide resistance and malaria transmission indicators in Anopheles gambiae s. l. in Bobo – Dioulasso, Burkina Faso : implications for vector control strategies. (2025).
Santolamazza, F. et al. Insertion polymorphisms of SINE200 retrotransposons within speciation Islands of Anopheles Gambiae molecular forms. Malar. J. 7, 163 (2008).
Hien, D. F. D. S. et al. Contrasting effects of the alkaloid ricinine on the capacity of Anopheles Gambiae and Anopheles coluzzii to transmit Plasmodium falciparum. Parasites Vectors. 14, 1–11 (2021).
Crawley, M. J. The R Book (John Wiley & Sons Ltd., 2007).
Gnémé, A. et al. Equivalent susceptibility of Anopheles Gambiae M and S molecular forms and Anopheles arabiensis to Plasmodium falciparum infection in Burkina Faso. Malar. J. https://doi.org/10.1186/1475-2875-12-204 (2013).
Ndiath, M. O. et al. Methods to collect Anopheles mosquitoes and evaluate malaria transmission: A comparative study in two villages in Senegal. Malar. J. 10, 270 (2011).
Ayo, D. et al. Susceptibility of Anopheles Gambiae to natural Plasmodium falciparum infection: A comparison between the Well-Established Anopheles Gambiae s.s line and a newly established Ugandan Anopheles Gambiae s.s. line. Am. J. Trop. Med. Hyg. https://doi.org/10.4269/ajtmh.23-0203 (2024).
Angleró-Rodríguez, Y. I. et al. A natural Anopheles -associated Penicillium chrysogenum enhances mosquito susceptibility to Plasmodium infection. Sci. Rep. https://doi.org/10.1038/srep34084 (2016).
Wang, M. et al. Glucose-mediated proliferation of a gut commensal bacterium promotes Plasmodium infection by increasing mosquito midgut pH. Cell. Rep. https://doi.org/10.1016/j.celrep.2021.108992 (2021).
Huang, W., Wang, S. & Jacobs-Lorena, M. Use of microbiota to fight Mosquito-Borne disease. Front. Genet. 11, 1–6 (2020).
Dong, Y., Manfredini, F. & Dimopoulos, G. Implication of the mosquito midgut microbiota in the defense against malaria parasites. PLoS Pathog. 5, 10 (2009).
Meister, S. et al. Anopheles Gambiae PGRPLC-mediated defense against bacteria modulates infections with malaria parasites. PLoS Pathog. 5, e1000542 (2009).
Cirimotich, C. M. et al. Natural microbe-mediated refractoriness to Plasmodium infection in Anopheles Gambiae. Science 332, 855–858 (2011).
Tchioffo, M. T. et al. Modulation of malaria infection in Anopheles Gambiae mosquitoes exposed to natural midgut bacteria. PLoS One. 8, 4–12 (2013).
Wang, S. & Jacobs-Lorena, M. Genetic approaches to interfere with malaria transmission by vector mosquitoes. Trends Biotechnol. 31, 185–193 (2013).
Blanford, S. et al. Fungal pathogen reduces potential for malaria transmission. Sci. (80-). 308, 1638–1641 (2005).
Bargielowski, I. & Koella, J. C. A possible mechanism for the suppression of Plasmodium Berghei development in the mosquito Anopheles Gambiae by the microsporidian Vavraia culicis. PLoS One 4, (2009).
Aliota, M. T., Chen, C. C., Dagoro, H., Fuchs, J. F. & Christensen, B. M. Filarial worms reduce plasmodium infectivity in mosquitoes. PLoS Negl. Trop. Dis. 5, e963 (2011).
Maïga, H., Dabiré, R. K., Lehmann, T., Tripet, F. & Diabaté, A. Variation in energy reserves and role of body size in the mating system of Anopheles Gambiae. J. Vector Ecol. 37, 289–297 (2012).
Sawadogo, S. P. et al. Effects of age and size on Anopheles Gambiae s.s. Male mosquito mating success. J. Med. Entomol. https://doi.org/10.1603/ME12041 (2013).
Zhou, G. et al. Metabolic fate of [ 14 C ] -labeled meal protein amino acids in Aedes aegypti mosquitoes. 50, 337–349 (2004).
Hansen, I. A., Attardo, G. M., Park, J., Peng, Q. & Raikhel, A. S. Target of rapamycin-mediated amino acid signaling in mosquito anautogeny. (2004).
Stone, C. M., Jackson, B. T. & Foster, W. A. Effects of plant-community composition on the vectorial capacity and fitness of the malaria mosquito Anopheles Gambiae. Am. J. Trop. Med. Hyg. 87, 727–736 (2012).
Funding
The research received financial support from Fundação para a Ciência e Tecnologia and Aga-Khan Development Network (MosqDiet_FCT AGAKHAN/541725581/2019), and IHI (Training and Capacity Building Unit). J.M. and H.S. are supported by the Fundação para a Ciência e a Tecnologia (FCT Portugal) by funds to GHTM/IHMT-NOVA (GHTM-UID GHTM UID/04413/2020). J.M. is recipient of a contract by FCT (https://doi.org/10.54499/CEECIND/00450/2017/CP1415/CT0001.
Author information
Authors and Affiliations
Contributions
H.S., J.M., R.V., I.S., E.B and D.F.S.H conceived and designed the present study; J.M. and R.V. produced the diet. D.F.S.H, A.M.A.P.S, and I.S. conducted the experiments, with E.B. providing supervision and overseeing the study; B.S.P. and D.F.S.H carried out the data analysis; D.F.S.H drafted the manuscript; E.B., I.S., H.S., J.M., A.D and R.V. provided critical revisions to the manuscript. All authors reviewed and approved the final version of the manuscript.
Corresponding authors
Ethics declarations
Ethics approval and consent to participate
The protocol of the study was approved by the Institut de Recherche en Sciences de la Santé, ethics committee: A 009-2023/CEIRES/IRSS. Prior to inclusion, informed consent was obtained from the parents or legal guardian of the volunteers.
Consent for publication
All authors concur with the submission presented by the corresponding authors.
Competing interests
The authors declare no competing interests.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Supplementary Material 1
Supplementary Material 2
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. 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-nc-nd/4.0/.
Reprints and permissions
About this article
Cite this article
de Sales Hien, D.F., Sare, I., Sib, A.M.A.P. et al. Effects of a bloodless diet on fitness and malaria susceptibility in Anopheles mosquitoes from Burkina Faso.
Sci Rep (2026). https://doi.org/10.1038/s41598-026-40458-x
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-40458-x
Keywords
- Bloodless diet
- Wing length
- Susceptibility to Plasmodium falciparum
- Longevity, Anopheles mosquitoes
Source: Ecology - nature.com
