title: The persistence and ecological impacts of a cyanobacterium genetically engineered to express mosquitocidal Bacillus thuringiensis toxins.
abstract: The cyanobacterium Anabaena PCC 7120#11 has been genetically engineered to act as a delivery vehicle for Bacillus thuringiensis subspecies israelensis mosquitocidal toxins. To address ecological concerns about releasing this genetically engineered microorganism into the environment for mosquito larva control, the persistence and ecological impacts of PCC 7120#11 was evaluated using multi-species, standardized aquatic microcosms.
  
  
  
  The microcosms were set up as described in ASTM E1366-02 (Standard Practice for Standardized Aquatic Microcosms: Fresh Water), with a few modifications. The treatment group microcosms were inoculated with PCC 7120#11 and key water quality parameters and non-target effects were compared between the treatment and control groups over a period of 35 days.
  
  
  
  PCC 7120#11 decreased from a concentration of 4.50 × 10(6) cells/ml (at inoculation) to 1.32 × 10(3) cells/ml after 4 weeks and larvicidal activity against third instar larvae of Anopheles arabiensis was only evident for two weeks after treatment. Both treatment and the interaction of treatment and time had a significant effect on nitrate, phosphate and photosynthetic microorganism concentrations. Treatment with PCC 7120#11 caused a temporary spike in ammonia in the microcosms a week after treatment, but the concentrations were well below acute and chronic criteria values for ammonia in freshwater ecosystems. Cyprinotus vidua concentrations were not significantly different between PCC 7120#11 and control microcosms. In PCC 7120#11 microcosms, Daphnia pulex concentrations were significantly lower than control concentrations between days 18 and 25. By the end of the experiment, none of the measured variables were significantly different between the treatment groups.
  
  
  
  The standard aquatic microcosm experiments provided more data on the ecological impacts of PCC 7120#11 than single-organism assessments would have. On the basis of the relatively minor, short-term effects that PCC 7120#11 had on water quality parameters and non-target invertebrates, further evaluation of PCC 7120#11 for use in integrated vector management is warranted.
authors.affiliation: School of Molecular and Cell Biology, University of the Witwatersrand, , Private Bag 3, Wits, 2050, Johannesburg, South Africa.
authors.forename: Irene
authors.initials: I
authors.lastname: Ketseoglou
authors.affiliation: School of Molecular and Cell Biology, University of the Witwatersrand, , Private Bag 3, Wits, 2050, Johannesburg, South Africa. gustav.bouwer@wits.ac.za.
authors.forename: Gustav
authors.initials: G
authors.lastname: Bouwer
mesh_terms: D017033:Anabaena / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D000852:Anopheles / Q000502:physiology*; D001413:Bacillus thuringiensis / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D017753:Ecosystem; D004731:Endotoxins / Q000235:genetics* / Q000378:metabolism; D007448:Invertebrates / Q000254:growth & development; D007814:Larva; D009032:Mosquito Control; D009584:Nitrogen / Q000032:analysis; D030781:Organisms, Genetically Modified; D010710:Phosphates / Q000032:analysis; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014867:Water / Q000592:standards*; D014871:Water Microbiology
chemical_list: C029103:Bacillus thuringiensis protoxin; D001427:Bacterial Toxins; D004731:Endotoxins; D010710:Phosphates; D011498:Protein Precursors; D014867:Water; D009584:Nitrogen
keywords: Anabaena PCC 7120; Anopheles arabiensis; Bacillus thuringiensis israelensis; Cry proteins; Genetically modified organism; Microcosms; Non-target organisms