title: Toxicity of Bacillus thuringiensis to parasitic and free-living life-stages of nematode parasites of livestock.
abstract: A collection of Bacillus thuringiensis (Bt) strains (Bts) were screened for activity against the free-living larval stages of nematode parasites of livestock. Two strains were identified with significant activity in inhibiting larval development of Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia circumcincta. These strains were also toxic to the adult parasitic stages of these nematode species in vitro. Adult H. contortus and O. circumcincta showed complete cessation of movement within 2 and 4 days, respectively. Trichostrongylus colubriformis adults were less affected, however, movement was still significantly reduced compared with controls. The in vitro activity against the larval stages was of a magnitude similar to or greater than that seen with the anthelmintic drugs thiabendazole and levamisole. N-terminal amino acid sequencing indicated that the two Bts contained either Cry5A and Cry5B proteins, or a Cry13 protein, and the presence of the corresponding cry5A, cry5B and cry13 genes was confirmed by PCR and sequencing. Bacillus thuringiensis spore-crystal suspensions exposed to acidic pH conditions (pH<or=3) showed greatly reduced toxicity in subsequent bioassays with nematode larvae, highlighting the need to protect the toxin from the acidic conditions of the sheep abomasum if it were to be administered per os as an anthelmintic. This study indicates that both the parasitic adult stages and the free-living larval stages of economically significant nematode parasites are susceptible to the effects of Bt, thus identifying this group of toxins as potential biocontrol agents.
authors.affiliation: CSIRO Livestock Industries, Queensland Bioscience Precinct, 306 Carmody Road, St Lucia, Qld 4067, Australia. andrew.kotze@csiro.au
authors.forename: A C
authors.initials: AC
authors.lastname: Kotze
authors.forename: J
authors.initials: J
authors.lastname: O'Grady
authors.forename: J M
authors.initials: JM
authors.lastname: Gough
authors.forename: R
authors.initials: R
authors.lastname: Pearson
authors.forename: N H
authors.initials: NH
authors.lastname: Bagnall
authors.forename: D H
authors.initials: DH
authors.lastname: Kemp
authors.forename: R J
authors.initials: RJ
authors.lastname: Akhurst
mesh_terms: D000818:Animals; D000871:Anthelmintics / Q000494:pharmacology; D001413:Bacillus thuringiensis / Q000737:chemistry / Q000472:pathogenicity*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D001427:Bacterial Toxins / Q000032:analysis / Q000235:genetics; D001681:Biological Assay / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004731:Endotoxins / Q000032:analysis / Q000235:genetics; D005243:Feces / Q000469:parasitology; D006189:Haemonchus / Q000187:drug effects / Q000382:microbiology / Q000502:physiology; D006460:Hemolysin Proteins; D006863:Hydrogen-Ion Concentration; D007814:Larva / Q000382:microbiology / Q000502:physiology; D009068:Movement; D009348:Nematoda / Q000187:drug effects / Q000382:microbiology* / Q000502:physiology; D009349:Nematode Infections / Q000469:parasitology / Q000662:veterinary; D010028:Ostertagia / Q000187:drug effects / Q000382:microbiology / Q000502:physiology; D010572:Pest Control, Biological / Q000379:methods; D012756:Sheep / Q000469:parasitology; D012757:Sheep Diseases / Q000469:parasitology; D014254:Trichostrongylus / Q000187:drug effects / Q000382:microbiology / Q000502:physiology
chemical_list: D000871:Anthelmintics; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis