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Identification of Rare Bacterial Pathogens by 16S rRNA Gene Sequencing and MALDI-TOF MS
Published on: July 11, 2016
Microbial associates of the southern mole cricket (Scapteriscus borellii) are highly pathogenic
Sudarshan K Aryal1, Derreck Carter-House2, Jason E Stajich2
1Department of Nematology, University of California, Riverside, CA 92521, USA.
Abstract:
We report the isolation and identification of seven bacterial strains and one fungal strain from dead and diseased Scapteriscus borellii mole crickets collected from a golf course in southern California. Using 16S and 18S rRNA gene sequence analysis we identified the microbes as Serratia marcescens (red), S. marcescens (white), S. marcescens (purple), Achromobacter xylosoxidans, Chryseobacterium sp., Ochrobactrum anthropi, Tsukamurella tryosinosolvens, and Beauveria bassiana. We performed a dose response curve for each of these cricket-associated microbial strains (except T. tryosinosolvens) and two other strains of S. marcescens (DB1140 and ATCC 13880). We found that all of these microbes except O. anthropi were highly pathogenic to D. melanogaster compared to the other strains of S. marcescens. Injecting the mole cricket associated strains of Serratia into flies killed all infected flies in ≤24h. For all other strains, the median time to death of injected flies varied in a dose-dependent manner. In vivo growth assessments of these microbes suggested that the host immune system was quickly overcome. We used disease tolerance curves to better understand the host-microbe interactions. Further studies are necessary to understand in mechanistic detail the virulence mechanisms of these mole cricket associated microbes and how this association may have influenced the evolution of mole cricket immunity.
Insights
Microbes isolated from diseased mole crickets, including Serratia marcescens and Beauveria bassiana, were highly pathogenic to fruit flies (Drosophila melanogaster). These findings highlight potential virulence mechanisms and host-pathogen interactions in insect immunity.
Area of Science:
- Entomology
- Microbiology
- Insect Pathology
Background:
- Mole crickets (Scapteriscus borellii) can be significant pests in turfgrass environments.
- Understanding the microbial agents associated with diseased insects is crucial for developing novel pest control strategies.
Purpose of the Study:
- To isolate and identify microbes from diseased mole crickets.
- To assess the pathogenicity of these microbes against a model insect, Drosophila melanogaster.
Main Methods:
- Isolation and identification of microbes using 16S and 18S rRNA gene sequencing.
- Pathogenicity testing via dose-response assays in Drosophila melanogaster.
- In vivo growth assessments and disease tolerance curve analysis.
Main Results:
- Seven bacterial strains (Serratia marcescens, Achromobacter xylosoxidans, Chryseobacterium sp., Ochrobactrum anthropi, Tsukamurella tryosinosolvens) and one fungal strain (Beauveria bassiana) were identified.
- Most isolated microbes, particularly Serratia marcescens strains, exhibited high pathogenicity towards Drosophila melanogaster.
- In vivo growth indicated rapid evasion of the host immune system.
Conclusions:
- Microbes associated with Scapteriscus borellii, especially Serratia species, possess significant virulence factors.
- These findings provide a basis for further investigation into insect-microbe interactions and potential biological control agents.
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