Progress 03/10/08 to 03/09/13
Outputs
Progress Report Objectives (from AD-416): 1. To explore improvements to formulation of biocontrol yeast for field application by growers. Types of formulations are wet formulations with additives (preservatives, nutritional supplements, surfactants for spreading) and dry formulations for delivery of viable yeast as a dry powder for tank mixing. 2. Evaluation of yeast longevity under different storage and application conditions: time and temperature. 3. Application for controlling l of other microbial pathogens: plant pathogenic fungi, microbial contaminants of crops (Salmonella, E. Coli) Approach (from AD-416): Objective 1. Protective additives will be added to wet and dry yeast formulations and evaluated for improved shelf-life under lab conditions. Spray trials in the field may be used to evaluate effects on durability on plant leaves and fruits. Surfactants will be in the tank mix for even spreading. Objective 2. Yeast durability in a fully diluted mixture will be evaluated through time and temperatures series in lab so the upper threshold that might be expected under grower conditions. Objective 3. Ability to control fungi on agricultural crops. evaluations will be done using greenhouse tests on plants and pathogens of significance in tree fruit-nut and vegetable crops. Salmonella is a particular problem on almond. Initial experiments may involve field sprays on almond followed by collection of nuts and inoculation with Salmonella in lab-greenhouse and monitor Salmonella population Non-pathogenic E. Coli strains applied to lettuce and spinach in the greenhouse followed by yeast spray applications to determine whether potential control is possible. The effectiveness of the yeast against additional fungi of agricultural importance will initially be evaluated on tomato and potato. Documents SCA with UC-Davis. Saprophytic yeast, Pichia anomala has been demonstrated to inhibit the growth of Aspergillus flavus as well as aflatoxin production by this fungus. It was also shown to inhibit Alternaria spp, Penicillium spp, Botrytis spp in lab assays. A stable liquid formulation of the biocontrol yeast product has been developed and used by spraying onto horticultural crops. Yeast cells can be stored at 40C for up to six month with very little change in viability. Field trials to control Alternaria disease in pistachio orchards have been conducted over the past four years. Significant control of A. alternata were observed in 2009. Nut cluster damage was also evaluated and treatment effects for this measure of A. alternata infection was highly significant (P=0.001). Cluster damage was also moderately correlated (r=0.47, P=0.002) with yield differences. Pichia anomala was tested as a biocontrol agent to reduce the survival and growth of attenuated E. Coli 0157:H7 strain on spinach. The experiments were carried out in a quarantined green house. A water suspension of P. anomala was applied to spinach 24 hr before inoculating E. coli. Spinach plants were harvested at day 4 and day 7 after the treatment. The population of E. coli on spinach was reduced by five fold in comparison to plants sprayed with the bacteria without yeasts. For the harvest mold experiment, in-hull almonds were used. The results indicated the nut samples contain high incidence of harvest molds and the yeast does have an inhibitory effect on the growth of harvest molds for in hull almond samples. It was also observed that the nut control samples contain fungal growth with sporulation while the fungal growth in the yeast control and the yeast treated samples has no visual evidence of sporulation. In the experiment with E. coli, we tested five replicates of almonds in dried hulls. The results indicates the dried in shell almonds had a lower infection rate of bacteria by the yeast treatment. The goal of the specific cooperative agreement is to develop practical applications of P. anomala to control pathogenic fungi and bacteria, which contributes directly to Objective 4 of the in-house project.
Impacts
(N/A)
Publications
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Progress 10/01/11 to 09/30/12
Outputs
Progress Report Objectives (from AD-416): 1. To explore improvements to formulation of biocontrol yeast for field application by growers. Types of formulations are wet formulations with additives (preservatives, nutritional supplements, surfactants for spreading) and dry formulations for delivery of viable yeast as a dry powder for tank mixing. 2. Evaluation of yeast longevity under different storage and application conditions: time and temperature. 3. Application for controlling l of other microbial pathogens: plant pathogenic fungi, microbial contaminants of crops (Salmonella, E. Coli) Approach (from AD-416): Objective 1. Protective additives will be added to wet and dry yeast formulations and evaluated for improved shelf-life under lab conditions. Spray trials in the field may be used to evaluate effects on durability on plant leaves and fruits. Surfactants will be in the tank mix for even spreading. Objective 2. Yeast durability in a fully diluted mixture will be evaluated through time and temperatures series in lab so the upper threshold that might be expected under grower conditions. Objective 3. Ability to control fungi on agricultural crops. evaluations will be done using greenhouse tests on plants and pathogens of significance in tree fruit-nut and vegetable crops. Salmonella is a particular problem on almond. Initial experiments may involve field sprays on almond followed by collection of nuts and inoculation with Salmonella in lab-greenhouse and monitor Salmonella population Non-pathogenic E. Coli strains applied to lettuce and spinach in the greenhouse followed by yeast spray applications to determine whether potential control is possible. The effectiveness of the yeast against additional fungi of agricultural importance will initially be evaluated on tomato and potato. Documents SCA with UC-Davis. Aspergillus flavus as well as aflatoxin production. A wet formulation has been developed by adding certain compounds in the formulation of yeast cells for storage at 40C for up to one year with very little change in viability. Chitosan was evaluated for its activity to enhance the biocontrol efficacy of the yeast, Pichia anomala against target fungal pathogens such as Aspergillus flavus. Yeast cells grew well at low concentrations of chitosan. The research is on going. The ADODR monitored this project through site visits, emails and phone calls. The goal of the specific cooperative agreement is to develop practical applications of Pichia anomala to control pathogenic fungi and bacteria, which contributes directly to Objective 4 of the in-house project. The SCA has been extended to March 2013.
Impacts
(N/A)
Publications
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Progress 10/01/10 to 09/30/11
Outputs
Progress Report Objectives (from AD-416) 1. To explore improvements to formulation of biocontrol yeast for field application by growers. Types of formulations are wet formulations with additives (preservatives, nutritional supplements, surfactants for spreading) and dry formulations for delivery of viable yeast as a dry powder for tank mixing. 2. Evaluation of yeast longevity under different storage and application conditions: time and temperature. 3. Application for controlling l of other microbial pathogens: plant pathogenic fungi, microbial contaminants of crops (Salmonella, E. Coli) Approach (from AD-416) Objective 1. Protective additives will be added to wet and dry yeast formulations and evaluated for improved shelf-life under lab conditions. Spray trials in the field may be used to evaluate effects on durability on plant leaves and fruits. Surfactants will be in the tank mix for even spreading. Objective 2. Yeast durability in a fully diluted mixture will be evaluated through time and temperatures series in lab so the upper threshold that might be expected under grower conditions. Objective 3. Ability to control fungi on agricultural crops. evaluations will be done using greenhouse tests on plants and pathogens of significance in tree fruit-nut and vegetable crops. Salmonella is a particular problem on almond. Initial experiments may involve field sprays on almond followed by collection of nuts and inoculation with Salmonella in lab-greenhouse and monitor Salmonella population Non-pathogenic E. Coli strains applied to lettuce and spinach in the greenhouse followed by yeast spray applications to determine whether potential control is possible. The effectiveness of the yeast against additional fungi of agricultural importance will initially be evaluated on tomato and potato. Saprophytic yeast, Pichia anomala has been demonstrated to inhibit the growth of Aspergillus flavus as well as aflatoxin production by this fungus. A stable liquid formulation is highly desirable because most of the biocontrol yeast products are dispersed in water and delivered by spraying or dipping to the agricultural and horticultural crops. A wet formulation has been developed. By adding certain compounds, the formulation of yeast cells can be stored at 40C for up to fourteen months with very little change in viability. The results were validated by flow cytometry analysis and fluorescent microscopy using viability stains. Pichia anomala was tested as a biocontrol agent to reduce the growth of A. flavus on almond fruits. A bioassay is being developed to screen compounds which can enhance the efficacy of yeast cells. The ADODR monitored this project through site visits, emails and phone calls. The goal of the specific cooperative agreement is to develop practical applications of P. anomla to control pathogenic fungi and bacteria, which contributes directly to Objective 4 of the in-house project.
Impacts
(N/A)
Publications
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Progress 10/01/09 to 09/30/10
Outputs
Progress Report Objectives (from AD-416) 1. To explore improvements to formulation of biocontrol yeast for field application by growers. Types of formulations are wet formulations with additives (preservatives, nutritional supplements, surfactants for spreading) and dry formulations for delivery of viable yeast as a dry powder for tank mixing. 2. Evaluation of yeast longevity under different storage and application conditions: time and temperature. 3. Application for controlling l of other microbial pathogens: plant pathogenic fungi, microbial contaminants of crops (Salmonella, E. Coli) Approach (from AD-416) Objective 1. Protective additives will be added to wet and dry yeast formulations and evaluated for improved shelf-life under lab conditions. Spray trials in the field will be used to evaluate effects on durability on plant leaves and fruits. Surfactants will be in the tank mix for even spreading. Objective 2. Yeast durability in a fully diluted mixture will be evaluated through time and temperatures series in lab so the upper threshold that might be expected under grower conditions. Objective 3. Ability to control fungi on agricultural crops. evaluations will be done using greenhouse tests on plants and pathogens of significance in tree fruit-nut and vegetable crops. Salmonella is a particular problem on almond. Initial experiments will involve field sprays on almond followed by collection of nuts and inoculation with Salmonella in lab-greenhouse and monitor Salmonella population. Non-pathogenic E. Coli strains applied to lettuce and spinach in the greenhouse followed by yeast spray applications to determine whether potential control is possible. The effectiveness of the yeast against additional fungi of agricultural importance will initially be evaluated on tomato and potato. Documents SCA with UC-Davis. Saprophytic yeast, Pichia anomala has been demonstrated to inhibit the growth of Aspergillus flavus as well as aflatoxin production by this fungus. It was also shown to inhibit Alternaria spp, Penicillium spp, Botrytis spp in lab assays. A stable liquid formulation is highly desirable because most of the bicontrol yeast products are dispersed in water and delivered by spraying or dipping to the agricultural and horticultural crops. A wet formulation has been developed. Yeast cells can be stored at 40C for up to six month with very little change in viability. We�ll monitor the stored yeast cells up to one year. Field trials to control Alternaria disease in pistachio orchards were conducted in the past four years. Significant control of A. alternata was observed in 2006 and 2009, but environmental conditions prevented A. alternata development in 2007 and 2008. Nut cluster damage was also evaluated and treatment effects for this measure of A. alternata infection was highly significant (P=0.001), due to the large number of error df for this study. Cluster damage was also moderately correlated (r=0.47, P=0.002) with yield differences. Pichia anomala was tested as a biocontrol agent to reduce the survival and growth of attenuated E. Coli 015:H7 strain on spinach. The experiments were carried out in a quarantined greenhouse. A water suspension of P. anomala was applied to spinach 24 hr before inoculating E. coli. Spinach plants were harvested at day 4 and day 7 after the treatment. The population of E. coli on spinach was reduced by five fold in comparison to plants sprayed with the bacteria without yeasts. The ADODR monitored this project through site visits, emails and phone calls. The goal of the specific cooperative agreement is to develop practical applications of P. anomala to control pathogenic fungi and bacteria, which contributes directly to Objective 2 of the in-house project.
Impacts
(N/A)
Publications
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Progress 10/01/08 to 09/30/09
Outputs
Progress Report Objectives (from AD-416) 1. To explore improvements to formulation of biocontrol yeast for field application by growers. Types of formulations are wet formulations with additives (preservatives, nutritional supplements, surfactants for spreading) and dry formulations for delivery of viable yeast as a dry powder for tank mixing. 2. Evaluation of yeast longevity under different storage and application conditions: time and temperature. 3. Application for controlling l of other microbial pathogens: plant pathogenic fungi, microbial contaminants of crops (Salmonella, E. Coli) Approach (from AD-416) Objective 1. Protective additives will be added to wet and dry yeast formulations and evaluated for improved shelf-life under lab conditions. Spray trials in the field will be used to evaluate effects on durability on plant leaves and fruits. Surfactants will be in the tank mix for even spreading. Objective 2. Yeast durability in a fully diluted mixture will be evaluated through time and temperatures series in lab so the upper threshold that might be expected under grower conditions. Objective 3. Ability to control fungi on agricultural crops. evaluations will be done using greenhouse tests on plants and pathogens of significance in tree fruit-nut and vegetable crops. Salmonella is a particular problem on almond. Initial experiments will involve field sprays on almond followed by collection of nuts and inoculation with Salmonella in lab-greenhouse and monitor Salmonella population. Non-pathogenic E. Coli strains applied to lettuce and spinach in the greenhouse followed by yeast spray applications to determine whether potential control is possible. The effectiveness of the yeast against additional fungi of agricultural importance will initially be evaluated on tomato and potato. Documents SCA with UC-Davis. Significant Activities that Support Special Target Populations Saprophytic yeast, Pichia anomala has been demonstrated to inhibit the growth of Aspergillus flavus as well as aflatoxin production by this fungus. It was also shown to inhibit Alternaria spp, Penicillium spp, Botrytis spp in lab assays. A stable liquid formulation is highly desirable because most of the bicontrol yeast products are dispersed in water and delivered by spraying or dipping to the agricultural and horticultural crops. A wet formulation to store the yeast cell in refrigerator for prolonged shelf life is in progress. The growth medium can produce 5 to 10 billions of cells per ml in a matter of three day. This amount of yeast cells can be diluted 100 to 1,000 fold for field sprays. Field trials are ongoing to control Alternaria disease in pistachio orchard. Because disease potential of Alternaria was low in 2008, the effect of Pichia anomala on Alternaria disease control could not be evaluated. A new strategy to flood the orchard floor will be adapted to create a humid environment in favor of Alternaria disease development. Two sprays of yeast cells were applied on June 11 and June 25, 2009, and two more sprays were planned for August, 2009. Pistachio nuts will be harvested in early September to evaluate nut yield. Alternaria disease symptom on pistachio trees will be examined. Pichia anomala was tested as a biocontrol agent to reduce the survival and growth of E. coli. Attenuated E. Coli 015:H7 strain was applied to spinach plants kept in a quarantined greenhouse. Preliminary results indicate that Pichia anomala reduced the number of E. coli on the spinach. Additional experiments are planned. The ADODR monitored this project through site visits, emails and phone calls.
Impacts
(N/A)
Publications
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Progress 10/01/07 to 09/30/08
Outputs
Progress Report Objectives (from AD-416) 1. To explore improvements to formulation of biocontrol yeast for field application by growers. Types of formulations are wet formulations with additives (preservatives, nutritional supplements, surfactants for spreading) and dry formulations for delivery of viable yeast as a dry powder for tank mixing. 2. Evaluation of yeast longevity under different storage and application conditions: time and temperature. 3. Application for controlling l of other microbial pathogens: plant pathogenic fungi, microbial contaminants of crops (Salmonella, E. Coli) Approach (from AD-416) Objective 1. Protective additives will be added to wet and dry yeast formulations and evaluated for improved shelf-life under lab conditions. Spray trials in the field will be used to evaluate effects on durability on plant leaves and fruits. Surfactants will be in the tank mix for even spreading. Objective 2. Yeast durability in a fully diluted mixture will be evaluated through time and temperatures series in lab so the upper threshold that might be expected under grower conditions. Objective 3. Ability to control fungi on agricultural crops. Evaluations will be done using greenhouse tests on plants and pathogens of significance in tree fruit-nut and vegetable crops. Salmonella is a particular problem on almond. Initial experiments will involve field sprays on almond followed by collection of nuts and inoculation with Salmonella in lab-greenhouse. and monitor Salmonella population. Non-pathogenic E. Coli strains applied to lettuce and spinach in the greenhouse followed by yeast spray applications to determine whether potential control is possible. The effectiveness of the yeast against additional fungi of agricultural importance will initially be evaluated on tomato and potato. Documents SCA with UC-Davis. Significant Activities that Support Special Target Populations Saprophytic yeast, Pichia anomala has been demonstrated to inhibit the growth of Aspergillus flavus as well as aflatoxin production by this fungus. It was also shown to inhibit Alternaria spp, Penicilum spp, Botrytis spp in lab assays. Formulation of biocontrol yeast is very important for prolonging the shelf-life of the yeast product and enhancing biocontrol activity in field application. A wet formulation to store the yeast cell pastes at 40C in saline and glycerol solution has been used for field experiments. Additives such as lactose, glycerol, and trehalose were demonstrated to prolong the shelf life of yeast cells in wet paste. Encapsulation of the yeast cells in bio-polymer is in progress. Yeast cells suspended in tank water were monitored for viability by determine the CFU (colony forming unit) on nutrient agar. The viability of yeast cells was not affected at 300C for several hours in water suspension. There was slight decline in viability when the tank water temperature was 350C in 2 hours. Field trials are ongoing to control Alternaria disease in pistachio orchard. Air samples were taken after yeast spray in orchard. Preliminary results showed the sprayed yeast cells were not detected in the surrounding air. Green-house experiments are being set up at UC Davis to control Alternaria spp infecting pistachio, potato and tomato by Pichia anomala. The ADODR monitored this project through site visits, emails and phone calls.
Impacts
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Publications
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