title: Neem oil increases the efficiency of the entomopathogenic fungus Metarhizium anisopliae for the control of Aedes aegypti (Diptera: Culicidae) larvae.
abstract: BACKGROUND
  Entomopathogenic fungi are potential candidates for use in integrated vector management and many isolates are compatible with synthetic and natural insecticides. Neem oil was tested separately and in combination with the entomopathogenic fungus Metarhizium anisopliae against larvae of the dengue vector Aedes aegypti. Our aim was to increase the effectiveness of the fungus for the control of larval mosquito populations.
  
  
  METHODS
  Commercially available neem oil was used at concentrations ranging from 0.0001 to 1%. Larval survival rates were monitored over a 7 day period following exposure to neem. The virulence of the fungus M. anisopliae was confirmed using five conidial concentrations (1 × 10(5) to 1 × 10(9) conidia mL(-1)) and survival monitored over 7 days. Two concentrations of fungal conidia were then tested together with neem (0.001%). Survival curve comparisons were carried out using the Log-rank test and end-point survival rates were compared using one-way ANOVA.
  
  
  RESULTS
  1% neem was toxic to A. aegypti larvae reducing survival to 18% with S50 of 2 days. Neem had no effect on conidial germination or fungal vegetative growth in vitro. Larval survival rates were reduced to 24% (S50 = 3 days) when using 1 × 10(9) conidia mL(-1). Using 1 × 10(8) conidia mL(-1), 30% survival (S50 = 3 days) was observed. We tested a "sub-lethal" neem concentration (0.001%) together with these concentrations of conidia. For combinations of neem + fungus, the survival rates were significantly lower than the survival rates seen for fungus alone or for neem alone. Using a combination of 1 × 10(7) conidia mL(-1) + neem (0.001%), the survival rates were 36%, whereas exposure to the fungus alone resulted in 74% survival and exposure to neem alone resulted in 78% survival. When using 1 × 10(8) conidia mL(-1), the survival curves were modified, with a combination of the fungus + neem resulting in 12% survival, whilst the fungus alone at this concentration also significantly reduced survival rates (28%).
  
  
  CONCLUSIONS
  The use of adjuvants is an important strategy for maintaining/increasing fungal virulence and/or shelf-life. The addition of neem to conidial suspensions improved virulence, significantly reducing larval survival times and percentages.
authors.affiliation: Department of Entomology and Plant Pathology, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, RJ, CEP 28013-602, Brazil. simoneazgomes@yahoo.com.br.
authors.forename: Simone A
authors.initials: SA
authors.lastname: Gomes
authors.affiliation: Department of Entomology and Plant Pathology, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, RJ, CEP 28013-602, Brazil. biodepaula@yahoo.com.br.
authors.forename: Adriano R
authors.initials: AR
authors.lastname: Paula
authors.affiliation: Department of Entomology and Plant Pathology, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, RJ, CEP 28013-602, Brazil. anderson.ribeirorj@yahoo.com.br.
authors.forename: Anderson
authors.initials: A
authors.lastname: Ribeiro
authors.affiliation: Department of Entomology and Plant Pathology, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, RJ, CEP 28013-602, Brazil. catiadepaula@gmail.com.
authors.forename: Catia O P
authors.initials: CO
authors.lastname: Moraes
authors.affiliation: Department of Entomology and Plant Pathology, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, RJ, CEP 28013-602, Brazil. jonathan_bastos@outlook.com.
authors.forename: Jonathan W A B
authors.initials: JW
authors.lastname: Santos
authors.affiliation: Departamento de Bioquímica, Universidade Federal de Santa Catarina, 88040-900, Florianópolis, Brazil. capsilva@ccb.ufsc.br.
authors.forename: Carlos P
authors.initials: CP
authors.lastname: Silva
authors.affiliation: Department of Entomology and Plant Pathology, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, RJ, CEP 28013-602, Brazil. richardiansamuels@gmail.com.
authors.forename: Richard I
authors.initials: RI
authors.lastname: Samuels
mesh_terms: D000330:Aedes / Q000187:drug effects* / Q000382:microbiology* / Q000502:physiology; D000818:Animals; D001681:Biological Assay; D005989:Glycerides / Q000494:pharmacology*; D007306:Insecticides / Q000494:pharmacology*; D007814:Larva / Q000187:drug effects / Q000382:microbiology / Q000502:physiology; D052981:Metarhizium / Q000187:drug effects / Q000254:growth & development* / Q000502:physiology; D009032:Mosquito Control / Q000379:methods*; D016019:Survival Analysis; D013729:Terpenes / Q000494:pharmacology*; D014774:Virulence / Q000187:drug effects
chemical_list: D005989:Glycerides; D007306:Insecticides; D013729:Terpenes; C002443:neem oil