Pathogenicity of microsclerotia from Metarhizium robertsii against Aedes aegypti larvae and antimicrobial peptides expression by mosquitoes during fungal-host interaction.

Aedes aegypti is a vector of various disease-causing arboviruses. Chemical insecticide-based methods for mosquito control have increased resistance in different parts of the world. Thus, alternative control agents such as the entomopathogenic fungi are excellent candidates to control mosquitoes as part of an ecofriendly strategy. There is evidence of the potential of entomopathogenic fungal conidia and blastospores for biological control of eggs, larval and adult stages, as well as the pathogenicity of fungal microsclerotia against adults and eggs. However, there are no studies on the pathogenicity of microsclerotia against either aquatic insects or insects that develop part of their life cycle in the water, such as the A. aegypti larvae. In this study, we assayed the production of microsclerotia and their pathogenicity against A. aegypti larvae of two isolates of Metarhizium robertsii, i.e., CEP 423 isolated in La Plata, Argentina, and the model ARSEF 2575. Both isolates significantly reduced the survival of A. aegypti exposed to their microsclerotia. The fungus-larva interaction resulted in a delayed response in the host. This was evidenced by the expression of some humoral immune system genes such as defensins and cecropin on the 9th day post-infection, when the fungal infection was consolidated as a successful process that culminates in larvae mortality. In conclusion, M. robertsii microsclerotia are promising propagules to be applied as biological control agents against mosquitoes since they produce pathogenic conidia against A. aegypti larvae.

Adulticidal Activity of Metarhizium anisopliae s.l. (Hypocreales: Clavicipitaceae) Native Strains From Argentina Against Aedes aegypti (Diptera: Culicidae).

The adulticidal activity of six fungal strains of Metarhizium anisopliae (Metschn.) Sorokin s.l. (Hypocreales: Clavicipitaceae) against Aedes aegypti (L.) (Diptera: Culicidae) was assessed. These strains (CEP 085, CEP 087, CEP 120, CEP 350, CEP 404, and CEP 423) were isolated from soil samples or nondipteran hosts collected from areas in Argentina where Ae. aegypti is distributed. Bioassays were performed with four conidial concentrations plus a control of each fungal strain to determine the lethal concentrations (LC50/LC90), the median survival times (ST50), the mean percentage of the surviving individuals, and the mean percentage of mycotized cadavers. The strains were able to infect and kill adult Ae. aegypti. Significant differences were found among the LC50 values, with CEP 423 as the most virulent strain with the lowest LC50 (2.4 × 106 conidia/ml). At 1 × 107 conidia/ml: no significant differences were found in the Kaplan-Meier survival functions among the strains; the ST50 ranged from 5 (CEP 085) to 8 d (CEP 350); and the mean percentage of the surviving adults was between 13.3 (CEP 085, CEP 350 and CEP 423) and 46.7% (CEP 087). Significant differences were also found among strains in the mean percentage of cadavers with fungal sporulation. Strain CEP 423 produced the highest percentage of mycotized adults (70%). The concentration of the inoculum significantly affected the survival of individuals and the percentage of mycotized cadavers within each strain. Metarhizium anisopliae s.l. CEP 423 was selected as the most promising candidate for further research aiming to develop a mycoinsecticide against Ae. aegypti.

Effect of UV-B Irradiation on Water-Suspended Metarhizium anisopliae s.l. (Hypocreales: Clavicipitaceae) Conidia and Their Larvicidal Activity in Aedes aegypti (Diptera: Culicidae).

Ultraviolet (UV) radiation is a key limiting factor for biological pest control with entomopathogenic fungi. While little is known about the impact of UV on Metarhizium anisopliae Metchnikoff (Sorokin) (Hypocreales: Clavicipitaceae) conidia in aquatic mosquito-breeding sites, this study determined the effect of UV-B on the viability and virulence of M. anisopliae sensu lato (s.l.) strain IP 46 in the laboratory against Aedes aegypti (L.) (Diptera: Culicidae) larvae. Conidia were treated in cups under defined water depths (0, 1, 2, and 3 cm) to six different UV-B doses (0, 0.657, 1.971, 3.942, 7.884, 11.826, or 15.768 kJ m-2) at 27 ± 2°C. The ability of treated conidia to germinate up to 24 h postexposure on PDAY + benomyl + chloramphenicol medium at 25 ± 1°C was adversely affected by higher doses of UV-B radiation regardless of the water depth. Germination, however, did not fall below 70% regardless of the test conditions. In fact, conidial virulence against second-instar larvae was not affected by either the water depth (F3,84 = 0.3, P = 0.85) or any tested levels of UV-B radiation (F6,21 ≤ 1.2, P ≥ 0.39) including those distinctly higher than might be expected for tropical sites. These findings strengthen previous observations that IP 46 has significant potential for use against A. aegypti larvae, even when exposed to elevated UV-B irradiance levels in the small breeding sites that are common for this important vector.