Microsclerotial pellets of Metarhizium spp.: thermotolerance and bioefficacy against the cattle tick.

The cattle tick, Rhipicephalus microplus (Acari: Ixodidae), is a multi-billion dollar ectoparasite of global importance affecting beef and milk production. Submerged cultures of cosmopolitan entomopathogenic fungal species of the genus Metarhizium typically produce microsclerotia that provide both long-term survival and environmental resistance. Microsclerotia hold great potential as an unconventional active propagule to control this tick under laboratory and semi-field conditions. However, heat stress caused especially by elevated temperatures poses a critical environmental constraint for the successful development and efficacy of microsclerotia under tropical conditions. First, we screened six strains of Metarhizium anisopliae, Metarhizium robertsii and Metarhizium humberi for their ability to produce microsclerotia by submerged liquid cultivation. In addition, we assessed the biological fitness and bioefficacy of dried microsclerotial pellets under amenable (27 °C) and heat-stressed (32 °C) incubation against engorged adult females of R. microplus. Microsclerotia in pelletized formulation prepared with carriers based on diatomaceous earth and microcrystalline cellulose exhibited conidial production at different extents according to the fungal strain and the incubation temperature, but most strains displayed reduced sporogenesis when exposed to 32 °C. Engorged tick females exposed to sporulated microsclerotia from pelletized M. anisopliae CG47 or IP 119 had fewer number of hatching larvae in comparison to the control group, irrespective of the incubation temperature tested. The minimum dosage of microsclerotial pellets that effectively reduced hatchability of tick larvae was estimated to be 2 mg per plate (equivalent to 6.0 kg per hectare). Metarhizium microsclerotial pellets exhibited significant tolerance to 32 °C and pronounced acaricidal activity against this economically important ectoparasite of cattle, even under simulated environmental heat stress. KEY POINTS: • Heat stress affects conidial production by microsclerotia of most pelletized Metarhizium strains • Heat stress does not impair the acaricidal performance of pelletized microsclerotia • Pellet formulation of Metarhizium microsclerotia is a promising mycoacaricide.

Inorganic pellets containing microsclerotia of Metarhizium anisopliae: a new technological platform for the biological control of the cattle tick Rhipicephalus microplus.

This study was sought to devise pellets containing inorganic materials and microsclerotia of Metarhizium anisopliae strain IP 119 for biological control of Rhipicephalus microplus, the most economically important tick in Brazilian cattle industry. In addition, we evaluated the storage stability of the pellets, their tolerance to ultraviolet radiation (UV-B), and efficacy against ticks under laboratory conditions. Fungal microsclerotia were produced by liquid culture fermentation and mixed with pre-selected inorganic matrices: vermiculite powder, diatomaceous earth, and colloidal silicon dioxide (78:20:2, w/w/w). The microsclerotial pellets were then prepared by a two-stage process involving extrusion and spheronization. Pellet size averaged 525.53 ± 7.74 µm, with a sphericity index of 0.72 ± 0.01, while biomass constituents did not affect the wet mass properties. Conidial production from microsclerotial pellets upon rehydration ranged from 1.85 × 109 to 1.97 × 109 conidia g-1 with conidial viability ≥ 93%. Conidial production from pellets stored at 4 °C was invariable for up to 21 days. Unformulated microsclerotia and microsclerotial pellets were extremely tolerant to UV-B compared with aerial conidia. Engorged tick females exposed to conidia from sporulated pellets applied to soil samples and upon optimal rehydration exhibited shorter oviposition time length, shorter life span, and reduced number of hatched larvae. In summary, microsclerotial pellets of M. anisopliae IP 119 effectively suppressed R. microplus and showed outstanding UV-B tolerance in laboratory tests. Prospectively, this formulation prototype is promising for targeting the non-parasitic stage of this tick on outdoor pasture fields and may offer a novel mycoacaricide for its sustainable management. KEY POINTS: • Pellets with microsclerotia and inorganic materials are innovative for tick control. • Metarhizium microsclerotia show superior UV-B tolerance in relation to conidia. • Pellets of Metarhizium microsclerotia produce infective conidia against ticks.

Relative humidity impacts development and activity against Aedes aegypti adults by granular formulations of Metarhizium humberi microsclerotia.

The impact of ambient relative humidity (RH) on conidial production of Metarhizium humberi IP 46 microsclerotia (MS) formulated in pellets or granules was investigated, and a promising granular formulation was tested against Aedes aegypti adults to confirm its efficacy. Microcrystalline cellulose (MC) and diatomaceous earth (DE) or a combination of vermiculite (VE), DE and silicon dioxide (SD) were tested as carriers in granular formulations containing MS. A range of 93-96.5% RH was critical for fungal development, and at least 96.5-98.5% RH was required for high conidial production on pellets or granules. Conidial production was clearly higher on pellets and granules prepared with VE than MC as the main carrier. VE granules containing MS were highly active against A. aegypti adults. Most mosquitoes were killed within 6 days after treatment regardless of the exposure time of adults to the formulation (1 min-24 h) or ambient humidity (75 or >98%). Production of conidia on dead adults varied between 7.3 × 106 and 2.2 × 107 conidia/individual, when exposed to MS granules for 12 h and 1 min, respectively. Granular formulations containing VE as the main carrier and MS as the active ingredient of M. humberi have strong potential for use against A. aegypti. KEY POINTS: • High conidial production on granular microsclerotial formulations at >96.5% RH • Vermiculite is more appropriate as a carrier than microcrystalline cellulose • Granules with IP 46 microsclerotia are highly active against Aedes aegypti adults.