RESUMO
The slow action of fungi is one of the biggest challenges in using entomopathogenic fungi. A promising alternative to reduce the time of action is to combine conidia with extracellular enzymes. This study aimed to characterize the production of Pr1 subtilisin protease and lipases by Beauveria bassiana and Metarhizium anisopliae in different culture media and to evaluate the efficiency of the enzymatic treatment against Aphis gossypii and Spodoptera frugiperda. The isolates were cultivated in five different liquid cultures, and, after 7 days, the culture was filtered and centrifuged, and the activity of the Pr1 and lipases was measured. The fungi cultured in a Luria-Bertani broth medium had the highest activity of proteases and lipases. The mortality of A. gossypii nymphs treated with conidia 7 days after the treatment was 39% (JEF-410), 76.5% (JEF-492), 74.8% (ERL-836), and 70.9% (JEF-214). The B. bassiana JEF-410 supernatant combined with conidia increased the fungal virulence at day 5 and day 6 after treatment. When S. frugiperda larvae were treated with B. bassiana JEF-492 conidia combined with its supernatant, the time of infection was shorter compared to the larvae treated with conidia only. Once the supernatant was incubated at 37 °C, the relative activity decreased from 100% to 80% after 2 h and to 45% after 24 h. The results suggest that the supernatant of entomopathogenic fungi may be formulated and used as a biopesticide in an efficient strategy for the biological control of pests.
RESUMO
Nutritional factors exert significant influence on the growth of entomopathogenic fungi, one of the main agents employed commercially in the biological control of arthropods. Thus, the objective of this work is to optimize the culture medium and solid fermentation time for production of proteases and conidia of Metarhizium robertsii ARSEF 2575 and to evaluate the interference of riboflavin and salts on virulence and resistance to abiotic stress factors. In the first step, nine groups were separated: negative control, positive control, and seven supplementation groups: ammonium nitrate, ammonium chloride, potassium nitrate, sodium nitrate, ammonium sulfate, ammonium phosphate, urea. Sodium nitrate showed significant difference in protease production at the time of 20 days of solid fermentation. Then, different concentrations of sodium nitrate and riboflavin as supplement were evaluated. Response surface methodology demonstrated that riboflavin and sodium nitrate influence proteolytic activity and conidia production, but without synergism. Supplementation of the medium with the optimal concentration of sodium nitrate and riboflavin did not interfere with the germination of conidia without exposure to abiotic stress, but did increase the thermotolerance of conidia. The presence of riboflavin and sodium nitrate at optimal concentrations in the culture medium did not alter fungal virulence with and without exposure to heat stress, varying according to the presence or absence of the supernatant during exposure, evidencing that resistance to heat exposure is multifactorial and dependent on intra- and extracellular factors. Moreover, the supplementation increased the larvicidal activity of the supernatant against Aedes aegypti.
RESUMO
Brazil concentrates the largest number of primate species in the world. In the present study, an extensive literature review of ticks on New World wild monkeys has been carried out, demonstrating that between the years 1912 to 2018, 182 larvae, 137 nymphs and 31 adult ticks (10 males and 21 females) were collected on 78 primates (from 12 different species) in 28 distinct localities in the Brazilian territory. Additionally, examination of allotments of 11 tick collections of Brazil revealed that from 1919 to 2019, 93 larvae, 91 nymphs and 175 adult ticks (62 males and 113 females) were collected from 100 monkeys (among 20 different species) from 43 localities in distinct Brazilian biomes. Overall, 19 tick species were identified on wild primates in the country: Amblyomma aureolatum (Pallas, 1772), Amblyomma cajennense (Fabricius, 1787) sensu stricto, Amblyomma coelebs Neumann, 1899, Amblyomma dubitatum Neumann, 1899, Amblyomma geayi Neumann, 1899, Amblyomma incisum Neumann, 1906, Amblyomma longirostre (Koch, 1844), Amblyomma naponense (Packard, 1869), Amblyomma nodosum Neumann, 1899, Amblyomma ovale Koch, 1844, Amblyomma parkeri Fonseca & Aragão, 1952, Amblyomma romarioi Martins, Luz & Labruna, 2019, Amblyomma rotundatum Koch, 1844, Amblyomma sculptum Berlese, 1888, Haemaphysalis juxtakochi Cooley, 1946, Ixodes fuscipes Koch, 1844, Rhipicephalus microplus (Canestrini, 1888), Rhipicephalus sanguineus (Latreille, 1806) sensu lato,and Ornithodoros rostratus Aragão, 1911. The presence of A. incisum, A. naponense, A. nodosum, A. rotundatum and I. fuscipes on monkeys is recorded for the first time. Thisresearch is therefore a significant contribution to the knowledge of tick speciesassociated with non-human primates in the Neotropical region.