Article

Virulence of Mexican isolates of entomopathogenic fungi (Hypocreales: Clavicipitaceae) upon Rhipicephalus=Boophilus microplus (Acari: Ixodidae) larvae and the efficacy of conidia formulations to reduce larval tick density under field conditions.

Departamento de Agronomía, División de Ciencias de la Vida, Campus Irapuato-Salamanca, Universidad de Guanajuato, Carretera Irapuato-Silao, Apartado Postal 311, Irapuato, Guanajuato, CP 36500, Mexico.
Veterinary Parasitology (Impact Factor: 2.55). 03/2010; 170(3-4):278-86. DOI: 10.1016/j.vetpar.2010.02.037
Source: PubMed

ABSTRACT The first objective was laboratory evaluation of the virulence of 53 Mexican isolates of fungi against larvae of Rhipicephalus (Boophilus) microplus. Thirty-three isolates of Metarhizium anisopliae var. anisopliae (Metschnickoff) Sorokin (Hypocreales: Clavicipitaceae) and 20 isolates of Isaria (Paecilomyces) fumosorosea (fumosoroseus) (Wize) (Eurotiales: Trichomaceae) were tested on 7-day-old larvae under laboratory conditions. Larvae were immersed in a suspension containing 10(8)conidia/mL and the CL(50) values were estimated. Then, field tests were conducted to determine the efficacy of formulations of the isolate with the highest virulence. M. anisopliae (Ma 14 isolate) was formulated with four carriers: Tween, Celite, wheat bran, and Citroline (mineral oil) and applied on pasture beds of Cynodon plectostachyus (L.), at a dose of 2 x 10(9)CFU/m(2). In the first trial, M. anisopliae was applied on plots naturally infested with larvae; in the second trial, tick populations in the experimental plots were eliminated and then re-infested with 20,000 7-day-old larvae. In the laboratory, all M. anisopliae isolates infected larvae with a mortality range between 2 and 100%; also, 13 of 20 I. fumosorosea isolates caused mortality rates between 7 and 94%. In the first field trial, 14 days post-application, conidial formulations in Celite and wheat bran caused 67.8 and 94.2% population reduction, respectively. In the second trial, the Tween formulation caused the highest larval reduction, reaching up to 61% (28 days post-application). Wheat bran formulation caused 58.3% larval reduction (21 days post-application) and was one of the most effective. The carriers and emulsifiers have a large impact on the effectiveness of conidial formulations.

0 Bookmarks
 · 
170 Views
  • [Show abstract] [Hide abstract]
    ABSTRACT: In the present study, the subtilisin-like protease Pr1 activity of five Metarhizium anisopliae s.l. isolates was evaluated together with their virulence against Rhipicephalus microplus females. Bioassays were performed with engorged females, and Pr1 activity was evaluated with the chromogenic substrate N-succinyl-Ala-Ala-Pro-Phe ρ-nitroanilide. All isolates were virulent to R. microplus, although the isolates CG 629, CG 148 and CG 32 caused higher mortality (P ≤ 0.05) than isolates CG 112 and CG 347 after eight days. No mortality was observed in the control group. Enzymatic assays revealed that the protease activity was greater in isolates CG 629, CG 148 or CG 32 than in isolates CG 112 or CG 347 after cultivation for 48 and 72 h. Based on these results, we suggest that the Pr1 activity may be used as a biochemical tool for screening the virulence among M. anisopliae s.l. isolates, and then selecting promising candidates for biocontrol of R. microplus ticks.
    Biocontrol Science and Technology 01/2014; 24(2). · 0.73 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The present study evaluated, for the first time, the effect of the commercial formulation Metarril® SP Organic of Metarhizium anisopliae plus 10% mineral oil to control Rhipicephalus microplus in a pen study. Three groups were formed with six animals each: the first group was exposed to Metarril® plus 10% mineral oil and 1% Tween 80; the second group was exposed to sterile distilled water, mineral oil and Tween 80 (oil control group); and the third group received no treatment (control group). The fungal formulation contained 1 × 108 conidia ml−1. Each animal was sprayed with 3 L of formulation. Fallen ticks were counted daily and a sample of 20 engorged females per day was incubated for assessment of biological parameters. Throughout the study period, Metarril® oil-based formulation showed an efficacy ranging from 19.20% to 67.39% in comparison with the control group; and from 8.18% to 61.38% in comparison with the oil control group. The average efficacy of Metarril® oil-based formulation was 47.74% and 40.89% in comparison with control and oil control groups, respectively. Changes in the biological parameters of engorged R. microplus females were observed in the first three days after treatment, with a significant reduction in hatching percentage and egg production index. We concluded that Metarril® SP Organic plus 10% mineral oil was efficient against R. microplus in pen studies. However, further in vivo studies are required to increase the efficacy and to establish a protocol for the use of this product in the field against the cattle tick.
    Veterinary Parasitology 09/2014; · 2.55 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The present study aimed to evaluate the pathogenic potential of different Metarhizium anisopliae s.l. isolates and to determine whether differences in enzymatic activities of proteases, lipases and chitinases and infection with mycoviruses affect the control of Rhipicephalus microplus achieved by these fungal isolates. Engorged female ticks were exposed to fungal suspensions. The lipolytic and proteolytic activities in the isolates were evaluated using chromogenic substrates and the chitinolytic activity was determined using fluorescent substrates. A gel zymography was performed to determine the approximate size of serine proteases released by M. anisopliae isolates. To detect mycoviral infections, dsRNA was digested using both RNAse A and S1 endonuclease; samples were analyzed on an agarose gel. Four of the five isolates tested were infected with mycovirus; however, the level of control of R. microplus ticks achieved with the only isolate free of infection (isolate CG 347) was low. This finding suggests that mycoviral infection does not affect the virulence of fungi against ticks. Although all five isolates were considered pathogenic to R. microplus, the best tick control and the highest levels of enzymatic activity were achieved with the isolates CG 629 and CG 148. The in vitro activities of lipases, proteases and chitinases produced by M. anisopliae s.l. differed among isolates and may be related to their virulence.
    Veterinary Parasitology 02/2014; · 2.55 Impact Factor

Full-text (2 Sources)

Download
278 Downloads
Available from
May 21, 2014