Alimentary tract bacteria isolated and identified with API- 20E and molecular cloning techniques from Australian tropical fruit flies, Bactrocera cacuminata and B. tryoni
1Institute of Agricultural Technology, Walailak University, Nakhon Si Thammarat, 80161, Thailand
2Center for Agricultural Biotechnology (AG-BIO/PERDO-CHE), Thailand
3International Centre for the Management of Pest Fruit Flies, Griffith School of Environment, Griffith University,
Nathan campus, Nathan, Queensland, 4111, Australia
4Griffith School of Environment, Griffith University, Nathan campus, Nathan, Queensland, 4111, Australia
Abstract
Bacteria were isolated from the crop and midgut of field collected Bactrocera cacuminata (Hering) and Bactrocera tryoni (Froggatt) (Diptera: Tephritidae). Two methods were used, firstly isolation onto two types of bacteriological culture media (PYEA and TSA) and identification using the API-20E diagnostic kit, and secondly, analysis of samples using the 16S rRNA gene molecular diagnostic method. Using the API-20E method, 10 genera and 17 species of bacteria in the family Enterobacteriaceae were identified from cultures growing on the nutrient agar. The dominant species in both the crop and midgut were Citrobacter freundii, Enterobacter cloacae and Klebsiella oxytoca. Providencia rettgeri, Klebsiella pneumoniae ssp ozaenae and Serratia marcescens were isolated from B. tryoni only. Using the molecular cloning technique that is based on 16S rRNA gene sequences, five bacteria classes were dignosed – Alpha-, Beta-, Gamma- and Delta- Proteobacteria and Firmicutes – including five families, Leuconostocaceae, Enterococcaceae, Acetobacteriaceae, Comamonadaceae and Enterobacteriaceae. The bacteria affiliated with Firmicutes were found mainly in the crop while the Gammaproteobacteria, especially the family Enterobacteriaceae, was dominant in the midgut. This paper presents results from the first known application of molecular cloning techniques to study bacteria within tephritid species and the first record of Firmicutes bacteria in these flies.
Keywords: Bactrocera cacuminata; Bactrocera tryoni; PCR; 16S rRNA gene; Enterobacteriaceae; lactic acid bacteria
Correspondence:
anarit_taochan@yahoo.com,
b*d.drew@griffith.edu.au,
cjane.hughes@griffith.edu.au,
ds.vijay@griffith.edu.au,
echin_anuchit@hotmail.com, *Corresponding author
Associate Editor:Allen Cohen was editor of this paper.
Received: 23 February 2009 | Accepted: 10 October 2009 | Published: 12 August 2010
Copyright: This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.
ISSN: 1536-2442 | Volume 10, Number 131
Thaochan N, Drew RAI, Hughes JM, Vijaysegaran S, Chinajariyawong A2010. Alimentary tract bacteria isolated and identified with API-20E and molecular cloning techniques from Australian tropical fruit flies, Bactrocera cacuminata and B. tryoni. Journal of Insect Science 10:131, available online: insectscience.org/10.131
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