Journal of Mosquito Research 2024, Vol.14, No.2, 67-75 http://emtoscipublisher.com/index.php/jmr 72 Zafar et al. (2020) showed that a strategy to manage resistance using varieties with multiple different Bt toxin genes (pyramided cotton) and RNA interference (RNAi) technology has been proposed. This approach, called multi-gene stacking and silencing (MGPS), aims to control pests by simultaneously expressing multiple Bt toxins and RNAi to knock out key genes of pests. The MGPS approach can delay or prevent the development of pest resistance to Bt cotton by delivering high doses of Bt toxins and RNAi and complying with refuge requirements. Bonin et al. (2015) suggested regular susceptibility monitoring of mosquito populations, using bioassays and genetic analysis methods to assess the level of resistance to Bti. This includes the use of QTL (quantitative trait loci) and admixture analysis to explore genetic markers associated with Bti resistance. In this way, signs of resistance can be detected in a timely manner and control strategies can be adjusted. Nascimento et al. (2020) showed that Cyt1Aa toxin plays a key role in promoting the entry of Bin toxin or its BinA subunit into midgut cells of mosquito larvae that lack Bin toxin receptors. This synergistic effect provides new ideas for the development of improved strategies to combat insect resistance. 3.3 Technical and practical challenges faced by Bti applications In the process of widespread adoption of Bacillus thuringiensis var. israelensis (Bti) for mosquito control, we face multiple technical and practical challenges that affect not only the effectiveness of Bti, but also its use in public health and ecosystems. application sustainability. The continuity and stability of Bti products is a significant technical challenge. Although Bti is widely considered to be relatively safe for non-target organisms, its long-term residue in the environment may have unknown effects on ecological balance. For example, Gutierrez-Villagomez et al. (2021) showed that the impact of Bti on amphibians may be more complex than previously thought, especially changes in their intestinal microbial communities that may affect the health and ecological functions of these organisms. In addition, the study by Johnson et al. (2020) revealed that in a mixed mangrove-salt marsh system, high mangrove coverage may significantly reduce the deposition of Bti products and reduce its lethality to mosquito larvae, indicating that the effect of Bti application is affected by Strong influence of ecosystem structure. The practical challenges of Bti application cannot be ignored. Cost is an important factor, especially in areas with limited resources. Although Bti is a more environmentally friendly option relative to chemical pesticides, the economic cost of its production and application remains a limiting factor. Additionally, community acceptance and participation are critical to the success of Bti control programs. Effective community communication and education activities can increase public awareness of the benefits of Bti applications and increase their acceptance. Although the application of Bti in mosquito control has faced challenges, these challenges have also prompted the development of scientific research and technological innovation. Through continued research, it will be possible to develop more efficient and sustainable Bti application strategies. For example, the research by Nascimento et al. (2020) explored the synergistic effect of Bti and other microbial toxins, providing a scientific basis for the development of new and more effective mosquito control products. 4 Discussion and Outlook This study provides insight into the effectiveness and challenges of using Bacillus thuringiensis var. israelensis (Bti) to control mosquito larvae (Aedes spp.) in aquaculture environments. The widespread application of Bti not only significantly improves public health and safety, reducing the risk of disease transmission by effectively reducing mosquito populations, but also highlights its important role in environmental protection, especially in reducing the use of chemical pesticides and protecting non-target Biological aspects (Brühl et al., 2020). Although Bti has gained wide acceptance and application as a biological control method, research in recent years has revealed several key challenges. The first is the development of mosquito resistance to Bti, which poses a threat to the long-term effectiveness of Bti. In addition, the effects of Bti application in different ecosystems and its potential impacts on non-target organisms and the entire ecosystem have also raised concerns. For example, recent studies have raised the need for further evaluation of the long-term ecological impacts of Bti, emphasizing
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