Comparison of water, oils and emulsifiable adjuvant oils as formulating agents for Metarhizium anisopliae for use in control of Boophilus microplus.

Studies were conducted to identify oil-based formulating agents (paraffinic oil, palm oil and emulsifiable adjuvant oils (EAOs)) for Metarhizium anisopliae that were superior to water with simple surfactants using a germination test and a bioassay against Boophilus microplus. Germination of conidia in all formulations, except 10% coconut EAO, produced more than 68% germination at 24 h and nearly 100% at 48 h. Coconut oil (average survival time (AST) = 4.6 +/- 0.28 days) and 10% liquid paraffin EAO (AST = 4.4 +/- 0.15 days) enhanced the pathogenicity of M. anisopliae to B. microplus relative to water (AST = 8.4 +/- 0.42 days). M. anisopliae in 10% liquid paraffin EAO was the most effective formulation having a moderately high germination after 24 h and a low AST as well as a high AST in the control. In the second experiment, germination of conidia in 2% liquid paraffin EAO and 2% Cropspray was higher than in 2% Codacide oil at 24 h, however, all treatments reached 100% germination after 48 h. The ASTs of the EAO based M. anisopliae formulations (Average AST = 6.4 +/- 0.54 days) were similar but lower that the ASTs of the controls (Average AST = 9.6 +/- 0.28 days).

Assessment of fungal isolates for development of a myco-acaricide for cattle tick control.

Entomopathogenic fungal isolates of Arachnid origin were assessed for their ability to produce mortality and inhibit egg hatching in Boophilus microplus with the aim of selecting an isolate for development into a myco-acaricide for control of cattle ticks. The ability of the most promising isolate to target developmental stages of more than one tick species and the optimum concentration of fungal inoculum to be used for future studies were determined. Metarhizium anisopliae was the most pathogenic of the three fungal species tested on B. microplus, producing shorter average survival times (ASTs) for engorged adults (AST = 5.2 +/- 0.1 days) and larvae (AST = 9.3 +/- 0.4 days), and a longer average hatching times (AHT; AHT = 19.8 +/- 0.5 days) in comparison to Simplicillium lamellicola and Paecilomyces farinosus. In comparative studies on two tick species with similar life cycles, M. anisopliae produced a shorter AST in engorged adult B. microplus (AST = 8.8 +/- 0.3 days) than Rhipicephalus sanguineus (AST = 10.3 +/- 0.3 days). M. anisopliae was pathogenic to larvae of B. microplus (AST = 7.7 +/- 0.4 days), however, had no effect on larvae of R. sanguineus (AST = 14.6 +/- 0.3 days) as the AST of this treatment was similar to its untreated control (AST = 14.1 +/- 0.4 days). M. anisopliae lengthened the AHTs in both B. microplus (AHT = 16.4 +/- 0.3 days) and R. sanguineus (AHT = 16.7 +/- 0.3 days) in comparison to the controls. The ASTs of engorged adult B. microplus treated with M. anisopliae shortened as the concentration was increased from 1 x 10(7) to 5 x 10(8) conidia/ mL. A further increase in concentration, 1 x 10(9) conidia/mL (AST = 10.2 +/- 0.4 days) did not shorten or lengthen the AST in comparison to 5 x 10(8) conidia/mL (AST = 9.4 +/- 0.3 days).

Thermal characteristics of Metarhizium anisopliae isolates important for the development of biological pesticides for the control of cattle ticks.

Experiments were conducted to determine if Metarhizium anisopliae isolates which are capable of growth at cattle surface temperatures could produce pathogenicity to Boophilus microplus in laboratory and field studies. The diurnal temperature fluctuation on the surface of cattle was monitored. The temperature tolerance of M. anisopliae isolates (ARSEF3297 and IMI386697) was determined and their pathogenicity to B. microplus compared at a standard bioassay temperature (28 degrees C) and at a temperature similar to the cattle surface (31-35 degrees C). The effect of the two isolates on the B. microplus population on cattle under field conditions was determined. The temperature of the fore udder, rear udder, ribs and neck regions of the mixed Holstein cattle fluctuated between 30 and 35 degrees C, in a similar pattern to the prevailing environmental temperature. However, wider fluctuations were obtained on the ears (28-35 degrees C) and spine (30-41 degrees C). The colony radius of both isolates declined as temperature increased, however, the growth of IMI386697 was five times greater than ARSEF3297 at 34 degrees C. At 28 degrees C, the pathogenicity of both isolates to B. microplus was similar, however, at 31-35 degrees C, IMI386697 was more pathogenic than ARSEF3297. Both isolates reduced the B. microplus population on cattle in comparison to the control formulation. However, IMI386697 (8.5+/-0.64 ticks/animal) produced a greater reduction in tick numbers than ARSEF3297 (19.1+/-0.64 ticks/animal). M. anisopliae was re-isolated from 8.9% of the ticks collected from IMI386697 treated cattle as compared to 2.8% of ticks from ARSEF3297 treated cattle.