The objective of this thesis was to investigate BiOWiSHTM-Aqua, a commercial dry solid formulation containing a consortium of bacteria and yeast, as a biopesticide for treatment of Botrytis cinerea, a gray mold that affects strawberries. BiOWiSHTM-Aqua was compared with another commercial product specifically used as a fungicide and bacteriocide, Serenade® Garden Disease Control Spray (concentrated Bacillus subtilis strain QST 713). Both laboratory tests as well as in vivo lab tests were conducted. BiOWiSHTM-Aqua results varied widely from plate to plate, regardless of experimental conditions. In some of these plates, inhibition zones were observed around colonies from BiOWiSHTM-Aqua, indicating efficacy. The organism responsible for the inhibition zones of B. cinerea growth was isolated from BiOWiSHTM-Aqua, and 16s rRNA analysis identified this culture as a strain of B. subtilis. This strain was designated as B. subtilis ssp. KLB. The B. subtilis KLB concentration required to completely inhibit B. cinerea was 9.1x104 CFU/mL when B. subtilis KLB was inoculated 48 hours before B. cinerea, 1.3x105 CFU/mL at 24 hours, and 3.2x106 CFU/mL when both were inoculated at the same time. Various preliminary experiments using B. subtilis KLB were also conducted to investigate its economic feasibility, to characterize the organism, and to test its post-harvest in vivo viability. B. subtilis KLB cell concentration was 1.6x109 CFU/mL in a bioreactor with LB at the end of the log growth phase. B. subtilis KLB achieved cell concentrations as high as 5x109 CFU/mL in shake flasks with food-grade tapioca as a carbon source. Inoculation of B. subtilis KLB on post-harvest strawberries did not have an effect on Botrytis infection rates compared to the negative control. These various experiments were the first step in research to potentially produce B. subtilis KLB on a commercial scale.
","abstract_html":"<p>The objective of this thesis was to investigate BiOWiSH<sup>TM</sup>-Aqua, a commercial dry solid formulation containing a consortium of bacteria and yeast, as a biopesticide for treatment of <em>Botrytis cinerea</em>, a gray mold that affects strawberries. BiOWiSH<sup>TM</sup>-Aqua was compared with another commercial product specifically used as a fungicide and bacteriocide, Serenade® Garden Disease Control Spray (concentrated <em>Bacillus subtilis</em> strain QST 713). Both laboratory tests as well as <em>in vivo</em> lab tests were conducted. BiOWiSH<sup>TM</sup>-Aqua results varied widely from plate to plate, regardless of experimental conditions. In some of these plates, inhibition zones were observed around colonies from BiOWiSH<sup>TM</sup>-Aqua, indicating efficacy. The organism responsible for the inhibition zones of <em>B. cinerea</em> growth was isolated from BiOWiSH<sup>TM</sup>-Aqua, and 16s rRNA analysis identified this culture as a strain of <em>B. subtilis</em>. This strain was designated as <em>B. subtilis</em> ssp. KLB. The <em>B. subtilis</em> KLB concentration required to completely inhibit <em>B. cinerea</em> was 9.1x10<sup>4</sup> CFU/mL when <em>B. subtilis</em> KLB was inoculated 48 hours before <em>B. cinerea</em>, 1.3x10<sup>5</sup> CFU/mL at 24 hours, and 3.2x10<sup>6</sup> CFU/mL when both were inoculated at the same time. Various preliminary experiments using <em>B. subtilis</em> KLB were also conducted to investigate its economic feasibility, to characterize the organism, and to test its post-harvest <em>in vivo</em> viability. <em>B. subtilis</em> KLB cell concentration was 1.6x10<sup>9</sup> CFU/mL in a bioreactor with LB at the end of the log growth phase. <em>B. subtilis</em> KLB achieved cell concentrations as high as 5x10<sup>9</sup> CFU/mL in shake flasks with food-grade tapioca as a carbon source. Inoculation of <em>B. subtilis</em> KLB on post-harvest strawberries did not have an effect on <em>Botrytis</em> infection rates compared to the negative control. These various experiments were the first step in research to potentially produce <em>B. subtilis</em> KLB on a commercial scale.</p>","abstract_has_math":false,"creators":["Ng, Kenneth K"],"institution":null,"degree_name":"MS in Engineering - Biochemical Engineering","degree_level":null,"degree_discipline":"Biomedical and General Engineering","degree_department":null,"school":null,"contributors":["Nirupam Pal"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2012,"date_issued":"2012-04-01T07:00:00Z","date_published":"2012-04-01T07:00:00Z","updated_at":"2026-07-24T01:31:11Z","subjects":["biopesticides","botrytis","bacillus subtilis","Agriculture","Biochemical and Biomolecular Engineering","Biotechnology","Environmental Microbiology and Microbial Ecology"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["10.15368/theses.2012.35"],"render_values":[{"text":"10.15368/theses.2012.35","href":"https://doi.org/10.15368/theses.2012.35","code":true}]}]},"links":{"outbound_url":"https://digitalcommons.calpoly.edu/theses/717","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Nirupam Pal"]},{"key":"dc:creator","label":"Author","values":["Ng, Kenneth K"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2014-04-24T07:00:00Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biomedical and General Engineering"]},{"key":"thesis:degree_name","label":"Degree Name","values":["MS in Engineering - Biochemical Engineering"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["biopesticides","botrytis","bacillus subtilis","Agriculture","Biochemical and Biomolecular Engineering","Biotechnology","Environmental Microbiology and Microbial Ecology"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://digitalcommons.calpoly.edu/theses/717","10.15368/theses.2012.35"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["The objective of this thesis was to investigate BiOWiSHTM-Aqua, a commercial dry solid formulation containing a consortium of bacteria and yeast, as a biopesticide for treatment of Botrytis cinerea, a gray mold that affects strawberries. BiOWiSHTM-Aqua was compared with another commercial product specifically used as a fungicide and bacteriocide, Serenade® Garden Disease Control Spray (concentrated Bacillus subtilis strain QST 713). Both laboratory tests as well as in vivo lab tests were conducted. BiOWiSHTM-Aqua results varied widely from plate to plate, regardless of experimental conditions. In some of these plates, inhibition zones were observed around colonies from BiOWiSHTM-Aqua, indicating efficacy. The organism responsible for the inhibition zones of B. cinerea growth was isolated from BiOWiSHTM-Aqua, and 16s rRNA analysis identified this culture as a strain of B. subtilis. This strain was designated as B. subtilis ssp. KLB. The B. subtilis KLB concentration required to completely inhibit B. cinerea was 9.1x104 CFU/mL when B. subtilis KLB was inoculated 48 hours before B. cinerea, 1.3x105 CFU/mL at 24 hours, and 3.2x106 CFU/mL when both were inoculated at the same time. Various preliminary experiments using B. subtilis KLB were also conducted to investigate its economic feasibility, to characterize the organism, and to test its post-harvest in vivo viability. B. subtilis KLB cell concentration was 1.6x109 CFU/mL in a bioreactor with LB at the end of the log growth phase. B. subtilis KLB achieved cell concentrations as high as 5x109 CFU/mL in shake flasks with food-grade tapioca as a carbon source. Inoculation of B. subtilis KLB on post-harvest strawberries did not have an effect on Botrytis infection rates compared to the negative control. These various experiments were the first step in research to potentially produce B. subtilis KLB on a commercial scale.
"]},{"key":"dc:title","label":"Title","values":["Investigation of Bacillus subtilis as a Biopesticide Against Botrytis cinerea"]}]}],"canonical_facts":{"dc:contributor":["Nirupam Pal"],"dc:creator":["Ng, Kenneth K"],"dc:date.available":["2014-04-24T07:00:00Z"],"dc:description.abstract":["The objective of this thesis was to investigate BiOWiSHTM-Aqua, a commercial dry solid formulation containing a consortium of bacteria and yeast, as a biopesticide for treatment of Botrytis cinerea, a gray mold that affects strawberries. BiOWiSHTM-Aqua was compared with another commercial product specifically used as a fungicide and bacteriocide, Serenade® Garden Disease Control Spray (concentrated Bacillus subtilis strain QST 713). Both laboratory tests as well as in vivo lab tests were conducted. BiOWiSHTM-Aqua results varied widely from plate to plate, regardless of experimental conditions. In some of these plates, inhibition zones were observed around colonies from BiOWiSHTM-Aqua, indicating efficacy. The organism responsible for the inhibition zones of B. cinerea growth was isolated from BiOWiSHTM-Aqua, and 16s rRNA analysis identified this culture as a strain of B. subtilis. This strain was designated as B. subtilis ssp. KLB. The B. subtilis KLB concentration required to completely inhibit B. cinerea was 9.1x104 CFU/mL when B. subtilis KLB was inoculated 48 hours before B. cinerea, 1.3x105 CFU/mL at 24 hours, and 3.2x106 CFU/mL when both were inoculated at the same time. Various preliminary experiments using B. subtilis KLB were also conducted to investigate its economic feasibility, to characterize the organism, and to test its post-harvest in vivo viability. B. subtilis KLB cell concentration was 1.6x109 CFU/mL in a bioreactor with LB at the end of the log growth phase. B. subtilis KLB achieved cell concentrations as high as 5x109 CFU/mL in shake flasks with food-grade tapioca as a carbon source. Inoculation of B. subtilis KLB on post-harvest strawberries did not have an effect on Botrytis infection rates compared to the negative control. These various experiments were the first step in research to potentially produce B. subtilis KLB on a commercial scale.
"],"dc:identifier":["https://digitalcommons.calpoly.edu/theses/717","10.15368/theses.2012.35"],"dc:subject":["biopesticides","botrytis","bacillus subtilis","Agriculture","Biochemical and Biomolecular Engineering","Biotechnology","Environmental Microbiology and Microbial Ecology"],"dc:title":["Investigation of Bacillus subtilis as a Biopesticide Against Botrytis cinerea"],"thesis:degree_discipline":["Biomedical and General Engineering"],"thesis:degree_name":["MS in Engineering - Biochemical Engineering"]},"updated_at":"2026-07-24T01:31:11Z"}