title: Molecular identification of hemolymph-associated symbiotic bacteria in red imported fire ant larvae.
abstract: The red imported fire ant (RIFA) (Solenopsis invicta Buren), an exotic insect pest in Texas, has become well established throughout the eastern part of the state. More aggressive than native ant species, RIFA gradually have enlarged their range and spread north and west despite intense efforts to stop them. Symbiotic bacteria have an important relationship in the midgut of fourth instar RIFA larvae. However, the presence of symbiotic bacteria in hemolymph has not been explored. In this study, symbiotic bacteria isolated from the hemolymph of fourth instar larvae of RIFA were genetically identified in terms of genus using a partial sequence of the 16S rRNA gene. Using three different primer sets to amplify regions of the gyrA, gyrB, and SG850 genes, multiple species of the genus Bacillus were identified as inhabitants of fire ant hemolymph. Analysis of gyrA gene identified Bacillus cereus with a percentage match of 94.13-99.20% with DNA sequences from GenBank BLAST ( www.ncbi.nih.gov ). Using the gyrB gene, Bacillus species were identified with a percentage match of 95.48% to 100% using DNA sequences from GenBank. Finally, analysis of the SG850 gene identified Bacillus cereus with a percentage match of 96.20% to 99.83% using DNA sequences from GenBank.
authors.affiliation: Department of Biology, The University of Texas at Tyler, 3900 University Blvd., Tyler, TX 75799, USA.
authors.forename: Stanley
authors.initials: S
authors.lastname: Gunawan
authors.forename: Danielle M
authors.initials: DM
authors.lastname: Tufts
authors.forename: Blake R
authors.initials: BR
authors.lastname: Bextine
mesh_terms: D000818:Animals; D001000:Ants / Q000382:microbiology*; D001407:Bacillus / Q000145:classification* / Q000302:isolation & purification*; D018407:Bacterial Physiological Phenomena; D027081:DNA Gyrase / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D020459:Genes, rRNA; D006458:Hemolymph / Q000382:microbiology*; D007814:Larva / Q000382:microbiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D013559:Symbiosis; D013781:Texas
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D027081:DNA Gyrase
references.citation: J Invertebr Pathol. 1977 Mar;29(2):192-200
references.pmid: 850074
references.citation: J Biosci Bioeng. 2001;92(4):342-5
references.pmid: 16233108
references.citation: Appl Environ Microbiol. 1999 Mar;65(3):1298-303
references.pmid: 10049896
references.citation: J Invertebr Pathol. 2005 Jul;89(3):203-9
references.pmid: 16039667
references.citation: J Antimicrob Chemother. 2004 Jul;54(1):90-4
references.pmid: 15190035
references.citation: FEMS Microbiol Ecol. 2006 Apr;56(1):132-40
references.pmid: 16542411