Whiteflies (Hemiptera: Aleyrodidae) are significant agricultural pests worldwide, primarily because they act as vectors for various plant viruses, including geminiviruses. Additionally, whitefly activity fosters the development of fungal colonies, such as sooty mold (Capnodium spp.), which grows on the sugary secretions (honeydew) produced by these insects. The presence of sooty mold reduces the plant’s photosynthetic capacity, further worsening the negative impact on its growth and development. Together, these factors make whitefly control a challenge for farmers, who must implement integrated pest management strategies to mitigate both the direct damage and the associated diseases that whiteflies can spread. Although approximately 1,200 species of whiteflies have been described, fewer than 40 are considered primary pests, with Bemisia tabaci (Gennadius, 1889) and Trialeurodes vaporariorum (Westwood, 1889) responsible for the greatest agricultural losses worldwide. The emergence of B. tabaci biotypes has prompted significant research, as these biotypes have been linked to economic damage to crops in several nations of the Americas, including Argentina and Brazil. In Paraguay, there is a need to identify local biotypes of B. tabaci led to the collection of whitefly samples from soybean plots in Capitán Miranda and Natalio, Itapúa, Paraguay. DNA was extracted using the Wizard genomic purification kit, followed by PCR amplification of the mitochondrial cytochrome oxidase I (mtCOI) region using specific primers for B. tabaci and T. vaporariorum. Agarose gel electrophoresis confirmed the presence of B and Q biotypes of B. tabaci in both Paraguayan locations, with no detection of T. vaporariorum. Sequencing of the PCR products revealed a 100% match with known sequences for B. tabaci and its biotypes, as confirmed by GenBank BLAST analysis. This research highlights the critical importance of continuously monitoring the population dynamics and geographical distribution of different biotypes. While the expansion of a polyphagous and dynamic pest like the whitefly cannot always be prevented, constant monitoring enables early detection of its spread and allows for the adjustment of management strategies tailored to each region. These measures will not eliminate the pest but will help mitigate damage and manage the problem sustainably and efficiently. The study also emphasizes the need to the effectiveness of insecticides, as resistance has been reported in other countries—an issue not yet explored in Paraguay.
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