Isolation of entomopathogenic fungi from soils and Ixodes scapularis (Acari

Ixodidae) ticks: Prevalence and methods

Amy Tuininga, Jessica L. Miller, Shannon U. Morath, Thomas J. Daniels, Richard C. Falco, Michael Marchese, Sadia Sahabi, Dieshia Rosa, Kirby C. Stafford

Research output: Contribution to journalArticleResearchpeer-review

25 Citations (Scopus)

Abstract

Entomopathogenic fungi are commonly found in forested soils that provide tick habitat, and many species are pathogenic to Ixodes scapularis Say, the blacklegged tick. As a first step to developing effective biocontrol strategies, the objective of this study was to determine the best methods to isolate entomopathogenic fungal species from field-collected samples of soils and ticks from an Eastern deciduous forest where I. scapularis is common. Several methods were assessed: (1) soils, leaf litter, and ticks were plated on two types of media; (2) soils were assayed for entomopathogenic fungi using the Galleria bait method; (3) DNA from internal transcribed spacer (ITS) regions of the nuclear ribosomal repeat was extracted from pure cultures obtained from soils, Galleria, and ticks and was amplified and sequenced; and (4) DNA was extracted directly from ticks, amplified, and sequenced. We conclude that (1) ticks encounter potentially entomopathogenic fungi more often in soil than in leaf litter, (2) many species of potentially entomopathogenic fungi found in the soil can readily be cultured, (3) the Galleria bait method is a sufficiently efficient method for isolation of these fungi from soils, and (4) although DNA extraction from ticks was not possible in this study because of small sample size, DNA extraction from fungi isolated from soils and from ticks was successful and provided clean sequences in 100 and 73% of samples, respectively. A combination of the above methods is clearly necessary for optimal characterization of entomopathogenic fungi associated with ticks in the environment.

Original languageEnglish
Pages (from-to)557-565
Number of pages9
JournalJournal of Medical Entomology
Volume46
Issue number3
DOIs
StatePublished - 1 May 2009

Fingerprint

Ixodidae
Ixodes
Ixodes scapularis
entomopathogenic fungi
Ticks
ticks
Acari
Fungi
Soil
Galleria
soil
methodology
DNA
plant litter
baits
fungi
isolation techniques
deciduous forests
internal transcribed spacers
Sample Size

Keywords

  • Biological control
  • Blacklegged tick
  • Entomopathogen
  • Fungi
  • Ixodes scapularis

Cite this

Tuininga, Amy ; Miller, Jessica L. ; Morath, Shannon U. ; Daniels, Thomas J. ; Falco, Richard C. ; Marchese, Michael ; Sahabi, Sadia ; Rosa, Dieshia ; Stafford, Kirby C. / Isolation of entomopathogenic fungi from soils and Ixodes scapularis (Acari : Ixodidae) ticks: Prevalence and methods. In: Journal of Medical Entomology. 2009 ; Vol. 46, No. 3. pp. 557-565.
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abstract = "Entomopathogenic fungi are commonly found in forested soils that provide tick habitat, and many species are pathogenic to Ixodes scapularis Say, the blacklegged tick. As a first step to developing effective biocontrol strategies, the objective of this study was to determine the best methods to isolate entomopathogenic fungal species from field-collected samples of soils and ticks from an Eastern deciduous forest where I. scapularis is common. Several methods were assessed: (1) soils, leaf litter, and ticks were plated on two types of media; (2) soils were assayed for entomopathogenic fungi using the Galleria bait method; (3) DNA from internal transcribed spacer (ITS) regions of the nuclear ribosomal repeat was extracted from pure cultures obtained from soils, Galleria, and ticks and was amplified and sequenced; and (4) DNA was extracted directly from ticks, amplified, and sequenced. We conclude that (1) ticks encounter potentially entomopathogenic fungi more often in soil than in leaf litter, (2) many species of potentially entomopathogenic fungi found in the soil can readily be cultured, (3) the Galleria bait method is a sufficiently efficient method for isolation of these fungi from soils, and (4) although DNA extraction from ticks was not possible in this study because of small sample size, DNA extraction from fungi isolated from soils and from ticks was successful and provided clean sequences in 100 and 73{\%} of samples, respectively. A combination of the above methods is clearly necessary for optimal characterization of entomopathogenic fungi associated with ticks in the environment.",
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Tuininga, A, Miller, JL, Morath, SU, Daniels, TJ, Falco, RC, Marchese, M, Sahabi, S, Rosa, D & Stafford, KC 2009, 'Isolation of entomopathogenic fungi from soils and Ixodes scapularis (Acari: Ixodidae) ticks: Prevalence and methods', Journal of Medical Entomology, vol. 46, no. 3, pp. 557-565. https://doi.org/10.1603/033.046.0321

Isolation of entomopathogenic fungi from soils and Ixodes scapularis (Acari : Ixodidae) ticks: Prevalence and methods. / Tuininga, Amy; Miller, Jessica L.; Morath, Shannon U.; Daniels, Thomas J.; Falco, Richard C.; Marchese, Michael; Sahabi, Sadia; Rosa, Dieshia; Stafford, Kirby C.

In: Journal of Medical Entomology, Vol. 46, No. 3, 01.05.2009, p. 557-565.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Isolation of entomopathogenic fungi from soils and Ixodes scapularis (Acari

T2 - Ixodidae) ticks: Prevalence and methods

AU - Tuininga, Amy

AU - Miller, Jessica L.

AU - Morath, Shannon U.

AU - Daniels, Thomas J.

AU - Falco, Richard C.

AU - Marchese, Michael

AU - Sahabi, Sadia

AU - Rosa, Dieshia

AU - Stafford, Kirby C.

PY - 2009/5/1

Y1 - 2009/5/1

N2 - Entomopathogenic fungi are commonly found in forested soils that provide tick habitat, and many species are pathogenic to Ixodes scapularis Say, the blacklegged tick. As a first step to developing effective biocontrol strategies, the objective of this study was to determine the best methods to isolate entomopathogenic fungal species from field-collected samples of soils and ticks from an Eastern deciduous forest where I. scapularis is common. Several methods were assessed: (1) soils, leaf litter, and ticks were plated on two types of media; (2) soils were assayed for entomopathogenic fungi using the Galleria bait method; (3) DNA from internal transcribed spacer (ITS) regions of the nuclear ribosomal repeat was extracted from pure cultures obtained from soils, Galleria, and ticks and was amplified and sequenced; and (4) DNA was extracted directly from ticks, amplified, and sequenced. We conclude that (1) ticks encounter potentially entomopathogenic fungi more often in soil than in leaf litter, (2) many species of potentially entomopathogenic fungi found in the soil can readily be cultured, (3) the Galleria bait method is a sufficiently efficient method for isolation of these fungi from soils, and (4) although DNA extraction from ticks was not possible in this study because of small sample size, DNA extraction from fungi isolated from soils and from ticks was successful and provided clean sequences in 100 and 73% of samples, respectively. A combination of the above methods is clearly necessary for optimal characterization of entomopathogenic fungi associated with ticks in the environment.

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KW - Biological control

KW - Blacklegged tick

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