Abstract
Entomopathogenic fungi, such as Metarhizium anisopliae, for the control of arthropods, have been studied for more than 20 years. The aim of this study was to determine the best methodology to evaluate the in vitro effect of the fungus M. anisopliae on Rhipicephalus microplus tick larvae. We compared a modified Larval Packet Test (LPT) and a Larval Immersion Test (LIT). For the LPT filter papers were impregnated with 1 mL of M. anisopliae suspension in Triton X-100 at 0.02%, in concentrations of 106, 107 and 108 conidia/mL and subsequently folded to include the larval ticks. LIT was performed by immersing the larvae in M. anisopliae suspensions for 5 min using the same three concentrations, then the larvae were placed on filter paper clips. For LPT, the LT50 values obtained were 134.6, 27.2 and 24.8 days for concentrations of 106, 107 and 108 conidia/mL; and the mortality after 21 days was 17.3, 17.6 and 38%, respectively. The LT50 values of LIT were 24.5, 20 and 9.2 days with mortality after 21 days of 50.5, 64.7 and 98% for 106, 107 and 108 conidia/mL, respectively. For the same conidia concentration, LIT showed a higher mortality in a shorter time interval when compared with LPT. These differences between the methods tested must be taking into account in further screening and effect studies with M. anisopliae. The set of results shown here could optimize the protocol used to identify M. anisopliae strains pathogenic against R. microplus.
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Acknowledgements
This study was financially supported by Fundação de Amparo a Pesquisa do Estado do Rio Grande do Sul (FAPERGS), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Conselho Nacional de Desenvolvimento Cientifico e Tecnológico (CNPq).
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Fig. 1
Larval mortality at different concentrations of Metharhizium anisopliae suspension. Panel A, Results of Larval Immersion Test (LIT). Panel B, Results of Larval Packet Test (LPT). The evaluation of tick mortality (counting the number of alive and dead larvae) was done throughout the 21 days of experiment daily. Spore suspension of M. anisopliae at 106 conidia/mL (■), 107 conidia/mL (▲), 108 conidia/mL (X) and control (♦). (TIFF 59 kb)
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Webster, A., Souza, U.A., Martins, J.R. et al. Comparative study between Larval Packet Test and Larval Immersion Test to assess the effect of Metarhizium anisopliae on Rhipicephalus microplus tick larvae. Exp Appl Acarol 74, 455–461 (2018). https://doi.org/10.1007/s10493-018-0235-1
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DOI: https://doi.org/10.1007/s10493-018-0235-1