title: No evidence for a culturable bacterial tetrodotoxin producer in Pleurobranchaea maculata (Gastropoda: Pleurobranchidae) and Stylochoplana sp. (Platyhelminthes: Polycladida).
abstract: Tetrodotoxin (TTX) is a potent neurotoxin found in the tissues of many taxonomically diverse organisms. Its origin has been the topic of much debate, with suggestions including endogenous production, acquisition through diet, and symbiotic bacterial synthesis. Bacterial production of TTX has been reported in isolates from marine biota, but at lower than expected concentrations. In this study, 102 strains were isolated from Pleurobranchaea maculata (Opisthobranchia) and Stylochoplana sp. (Platyhelminthes). Tetrodotoxin production was tested utilizing a recently developed sensitive method to detect the C9 base of TTX via liquid chromatography-mass spectrometry. Bacterial strains were characterized by sequencing a region of the 16S ribosomal RNA gene. To account for the possibility that TTX is produced by a consortium of bacteria, a series of experiments using marine broth spiked with various P. maculata tissues were undertaken. Sixteen unique strains from P. maculata and one from Stylochoplana sp. were isolated, representing eight different genera; Pseudomonadales, Actinomycetales, Oceanospirillales, Thiotrichales, Rhodobacterales, Sphingomonadales, Bacillales, and Vibrionales. Molecular fingerprinting of bacterial communities from broth experiments showed little change over the first four days. No C9 base or TTX was detected in isolates or broth experiments (past day 0), suggesting a culturable microbial source of TTX in P. maculata and Stylochoplana sp. is unlikely.
authors: Department of Biological Sciences, University of Waikato, Private Bag 3105, Hamilton 3240, New Zealand. ls161@students.waikato.ac.nz.
authors: Lauren R
authors: LR
authors: Salvitti
authors: Department of Biological Sciences, University of Waikato, Private Bag 3105, Hamilton 3240, New Zealand. paul.mcnabb@cawthron.org.nz.
authors: Susanna A
authors: SA
authors: Wood
authors: Cawthron Institute, Nelson 7042, New Zealand. paul.mcnabb@cawthron.org.nz.
authors: Paul
authors: P
authors: McNabb
authors: Department of Biological Sciences, University of Waikato, Private Bag 3105, Hamilton 3240, New Zealand. caryc@waikato.ac.nz.
authors: Stephen Craig
authors: SC
authors: Cary
mesh_terms: D000818:Animals; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D002853:Chromatography, Liquid; D013058:Mass Spectrometry; D059013:Microbial Consortia / Q000235:genetics / Q000502:physiology*; D010802:Phylogeny; D010986:Platyhelminths / Q000382:microbiology*; D049853:Pleurobranchaea / Q000382:microbiology*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013779:Tetrodotoxin / Q000032:analysis / Q000096:biosynthesis*
chemical_list: D012336:RNA, Ribosomal, 16S; D013779:Tetrodotoxin
references: Toxicol Mech Methods. 2002;12(2):135-53
references: 20021198
references: Mar Drugs. 2012 Jan;10(1):163-76
references: 22363228
references: Appl Environ Microbiol. 2006 Jul;72(7):5069-72
references: 16820507
references: Environ Toxicol Pharmacol. 2013 Nov;36(3):1046-54
references: 24121556
references: Mol Biol Evol. 1987 Jul;4(4):406-25
references: 3447015
references: Food Chem Toxicol. 1995 Nov;33(11):929-34
references: 7590540
references: J Bacteriol. 1998 Sep;180(18):4765-74
references: 9733676
references: Toxicon. 2008 Jul;52(1):55-61
references: 18573274
references: Appl Microbiol Biotechnol. 2002 Jul;59(2-3):125-34
references: 12111137
references: Toxicon. 2004 Sep 1;44(3):243-9
references: 15302530
references: Adv Food Nutr Res. 2007;52:141-236
references: 17425946
references: Appl Environ Microbiol. 1993 Nov;59(11):3934-7
references: 16349096
references: Toxicon. 2015 Mar;95:23-9
references: 25557071
references: Appl Environ Microbiol. 1995 Sep;61(9):3468-70
references: 7574655
references: Ann N Y Acad Sci. 1986;479:32-43
references: 3468846
references: Appl Environ Microbiol. 2004 Oct;70(10):6147-56
references: 15466561
references: Toxicon. 2010 Sep 1;56(3):324-9
references: 20362605
references: J Appl Microbiol. 2012 Aug;113(2):459-65
references: 22583619
references: Nature. 2000 Mar 23;404(6776):354
references: 10746714
references: Toxicon. 2004 Nov;44(6):641-7
references: 15501290
references: Mikrobiologiia. 2014 May-Jun;83(3):312-9
references: 25844441
references: Toxicon. 2007 Nov;50(6):779-90
references: 17698158
references: Nucleic Acids Res. 2004;32(4):1363-71
references: 14985472
references: Nat Prod Res. 2009;23(17):1630-40
references: 19851930
references: Toxicon. 2004 Dec 15;44(8):933-8
references: 15530976
references: Aquat Toxicol. 2011 Jul;104(1-2):61-72
references: 21543051
references: Angew Chem Int Ed Engl. 2014 Dec 22;53(52):14546-9
references: 25382791
references: Mar Drugs. 2013 Aug;11(8):2695-712
references: 23917066
references: Toxicon. 2005 Jun 1;45(7):851-9
references: 15904680
references: Environ Microbiol. 2006 May;8(5):929-38
references: 16623749
references: Mar Drugs. 2010;8(3):741-62
references: 20411124
references: Mar Drugs. 2011;9(11):2384-96
references: 22163191
references: J AOAC Int. 2014 Mar-Apr;97(2):325-33
references: 24830143
references: Mar Drugs. 2014 Jan;12(1):1-16
references: 24368566
references: Toxicon. 2005 Sep 15;46(4):471-6
references: 16051296
references: Appl Environ Microbiol. 1990 Apr;56(4):1162-3
references: 16348162
references: Toxicon. 1987;25(2):225-8
references: 3576639
references: Appl Environ Microbiol. 1985 Oct;50(4):781-90
references: 16346911
references: Appl Environ Microbiol. 2000 Apr;66(4):1698-701
references: 10742263
references: Toxicon. 2010 Sep 15;56(4):640-3
references: 20576513
references: J Biochem. 1986 Jan;99(1):311-4
references: 3754255
references: Toxicon. 2002 Mar;40(3):313-6
references: 11711129
references: Toxicon. 2010 Sep 1;56(3):466-73
references: 20466016