Sake CPA-1 Research Articles

Shelf life improvement of the biocontrol agent Candida sake CPA-1 by suitable package and storage conditions

Agrochemical companies are increasing their interest in the production of biopesticides based on biocontrol agents (BCAs), mainly due to the need to reduce the synthetic fungicide application and the consequent benefits for the environment and for human health. The efficacy of the BCA Candida sake CPA-1 has been demonstrated against the main postharvest diseases on pome fruit, and B. cinerea and sour rot in grapes control on field. Recently, two effective fluidised-bed spray-dried formulations based on C. sake CPA-1 and biodegradable compounds were described as an effective biocontrol product. However, the maintenance of the effectiveness and survival of these formulations during a long shelf life is a crucial aspect to the success of its marketing. For this reason, the aim of this study was to select the best packaging and storage conditions to maintain a long shelf life of two fluidised-bed spray-dried C. sake CPA-1 formulations. Viability and aw (water activity) of CPA-1 formulations stored in different packages (bottles and bags), temperatures (− 20 °C and 4 °C) and atmospheric conditions (vacuum and air) were tested for 22 months. Both formulations maintained their initial viability (around 4 × 109 CFU ml−1) when they were stored at − 20 °C regardless of the packages or atmospheric conditions. In contrast, the viability of CPA-1 formulations stored at 4 °C differed between bags and bottles. Both formulations could be stored for at least 21 months at − 20 °C, and only one formulation could achieve a one-year shelf life stored in bottles at 4 °C. C. sake CPA-1 formulations stored for 12 months at the most suitable conditions significantly reduced B. cinerea on grapes by 25 to 56%.

Verifying the biocontrol activity of novel film-forming formulations of Candida sake CPA-1: resilience in relation to environmental factors, rainfall episodes, and control of Botrytis cinerea on different hosts.

The efficacy of Candida sake CPA-1 as a biocontrol agent against several diseases has been studied since it was isolated 20 years ago. However, it was only recently that two suitable and effective film-forming formulations based on potato starch and maltodextrins were developed using the fluidized-bed spray-drying system. The present work aimed to confirm the capability of both novel formulations by testing their resilience on grapes at different temperatures (0, 22, and 30 °C), relative humidities (40% and 85%), and simulated rainfall levels. Another objective was to examine the control of Botrytis cinerea in different hosts. The CPA-1 cells from both dried formulations survived better than the liquid formulation on grapes stored at 0 and 22 °C regardless of the relative humidity. After simulated rainfall, potato starch formulation achieved significantly higher populations than maltodextrin formulation, although the highest reduction was -1.6 log N N0 -1 . A positive effect of cell establishment prior to the simulated rainfall was shown, and recovered cells from the potato starch formulation were significantly higher after 72 h of cell establishment. Finally, both formulations reduced the incidence and severity of B. cinerea on pears, apples, and tomatoes. The potential of these novel film-forming formulations of C. sake CPA-1 was verified. The resilience of formulated C. sake was better than the commercialized liquid formulation, the adherence of the formulations to the grapes improved after an establishment period prior to rain exposure, and the control of B. cinerea was verified in a wider range of hosts. © 2019 Society of Chemical Industry.

Predicted ecological niches and environmental resilience of different formulations of the biocontrol yeast Candida sake CPA-1 using the Bioscreen C

Environmental resilience of biocontrol microorganisms has been a major bottleneck in the development of effective formulations. Candida sake is an effective biocontrol agent (BCA) against Penicillium expansum, Botrytis cinerea or Rhizopus stolonifer, and different formulations of the BCA have been optimised recently. The objective of this study was to compare the relative tolerance of different dry and liquid formulations of the biocontrol yeast C. sake CPA-1 to interacting environmental conditions using the Bioscreen C. Initially, the use of this automated turbidimetric method was optimised for use with different formulations of the biocontrol yeast. The best growth curves were obtained for the C. sake CPA-1 strain when grown in a synthetic grape juice medium under continuous shaking and with an initial concentration of 105 CFUs ml−1. All the formulations showed a direct relationship between optical density values and yeast concentrations. Temperature (15–30 °C) and water activity (aw; 0.94–0.99) influenced the yeast resilience most profoundly, whereas the effect of pH (3–7) was minimal. In general, the liquid formulation grew faster in more interacting environmental conditions but only the yeast cells in the dry potato starch formulation could grow in some stress conditions. This rapid screening method can be used for effective identification of the resilience of different biocontrol formulations under interacting ecological abiotic conditions.

Novel film-forming formulations of the biocontrol agent Candida sake CPA-1: biocontrol efficacy and performance at field conditions in organic wine grapes.

The biocontrol agent (BCA) Candida sake CPA-1 has previously effectively reduced Botrytis bunch rot (BBR) and it was also suggested as a promising strategy to control sour rot in grapes under field conditions. However, biocontrol efficacy of solid formulations of CPA-1 has never been tested in field trials. The present study aims to confirm the efficacy against BBR and sour rot in grapes under field conditions of two novel formulations recently developed by the addition of biodegradable coatings using a fluidized-bed spray-drying system. Novel film-forming formulations of the BCA C. sake CPA-1 controlled B. cinerea as well as liquid formulation. Sour rot control resulted better in the second season and severity reductions were more satisfactory than incidence control. Visual and cryoSEM observations revealed that film-forming treatments were uniformly distributed on plant surfaces. CPA-1 coating could be observed on grapes at harvest time. The results of this work suggest that solid formulations would be a competitive alternative to conventional fungicides because they were easy to package and transport, and cell viability could be maintained for a long period of time. © 2018 Society of Chemical Industry.

Fluidised-bed spray-drying formulations of Candida sake CPA-1 by adding biodegradable coatings to enhance their survival under stress conditions.

The biocontrol agent Candida sake CPA-1 has demonstrated to be effective against several diseases on fruit. However, for application of CPA-1 under field conditions, it was necessary to mix it with a food coating to improve survival under stress conditions, as well as adherence and distribution on fruit surfaces. The objective of this study was to obtain a more competitive formulation under field conditions to be applied independently of any product. To achieve this purpose, the drying process of CPA-1 by a fluidised-bed spray-drying system together with biodegradable coatings was optimised. This approach is novel for the drying system used and the formulation obtained which was able to form a film or coating on fruit surfaces. Several substances were tested as carriers and binders, and drying temperature was optimised. The addition of protective compounds was also tested to improve survival of CPA-1 during the dehydration process. Product shelf life, biocontrol efficacy on grapes against Botrytis cinerea, and the improvement of C. sake behaviour under stress conditions were tested. The optimal temperature of drying was 55°C and two formulations that were able to develop a coating on fruit surfaces were obtained. One of the formulations was created by using a combination of native and pregelatinised potato starch; the other formulation was obtained using maltodextrin and by adding skimmed milk and sucrose as protectant compounds. The formulated products reduced the incidence and severity of B. cinerea, and CPA-1 survival rate was increased under stress conditions of temperature and humidity.

Dry formulations of the biocontrol agent Candida sakeCPA‐1 using fluidised bed drying to control the main postharvest diseases on fruits

The biocontrol agent Candida sake CPA-1 is effective against several diseases. Consequently, the optimisation of a dry formulation of C. sake to improve its shelf life and manipulability is essential for increasing its potential with respect to future commercial applications. The present study aimed to optimise the conditions for making a dry formulation of C. sake using a fluidised bed drying system and then to determine the shelf life of the optimised formulation and its efficacy against Penicillium expansum on apples. The optimal conditions for the drying process were found to be 40 °C for 45 min and the use of potato starch as the carrier significantly enhanced the viability. However, none of the protective compounds tested increased the viability of the dried cells. A temperature of 25 °C for 10 min in phosphate buffer was considered as the optimum condition to recover the dried formulations. The dried formulations should be stored at 4 °C and air-packaged; moreover, shelf life assays indicated good results after 12 months of storage. The formulated products maintained their biocontrol efficacy. A fluidised bed drying system is a suitable process for dehydrating C. sake cells; moreover, the C. sake formulation is easy to pack, store and transport, and is a cost-effective process. © 2017 Society of Chemical Industry.

Effective control of Botrytis bunch rot in commercial vineyards by large-scale application of Candida sake CPA-1

Biological control with microorganisms is regarded as a promising control strategy against Botrytis bunch rot (BBR, grey mould) on grapes. Candida sake CPA-1 is a yeast that has previously shown a high efficacy against BBR in small-scale field trials. The present work aims to confirm this efficacy in commercial conditions and to evaluate the compatibility of C. sake with phytosanitary products that are commonly used in viticulture. The large-scale spray programmes (two to three applications in the growing season) were carried out in three field sites in Catalonia, North-East Spain, with different vineyard management styles. The overall reductions were in the ranges of 35 ± 5.7 to 64 ± 3.7% incidence, and 39 ± 17.5 to 85 ± 3.9% severity, compared to the control. Only four out of 39 phytosanitary products, tested in liquid suspension with C. sake, were incompatible with the yeast. The results demonstrated the potential of C. sake to become a reliable solution as a biocontrol product to be integrated in the control of BBR within the future.

Stability of biocontrol products carrying Candida sake CPA-1 in starch derivatives as a function of water activity

ABSTRACTThe preservation and shelf-life of formulations of the biocontrol agent Candida sake CPA-1 and starch derivatives as a function of water activity (aW) were studied in terms of the physical stability of the products and cell viability. Formulations of biocontrol products (BCPs), based on combinations of potato starch and pre-gelatinised potato starch (F1 and F2) or maltodextrines (MD) (F3) containing cell protectants, were obtained by fluidised-bed drying. The carriers and the formulated products were stored at 20°C under different aW conditions. The water sorption and water plasticization behaviour of the different products were analysed through the water sorption isotherms and glass transition temperatures (Tg). Likewise, the viability of C. sake over time was determined as a function of the aW. The solubility of the products was also assessed. Although formulations stored at 20°C and low aW (≤ 0.33) exhibited a better shelf-life, a significant decrease in cell survival ratio after 180 storage days was observed. Cold storage (5°C) was required to better maintain the cell viability, thus prolonging the shelf-life of BCPs. Formulations containing MD were the most effective at preserving cell viability and also exhibited the highest water solubility. All the formulations were physically stable at ambient temperature; therefore, the cell stability is the critical point at which to establish both the aW levels and temperature during storage. Packaging the product using high water vapour barrier material and under cold storage would be necessary to ensure a high number of viable cells and an effective and competitive BCP.

Properties of biopolymer dispersions and films used as carriers of the biocontrol agent Candida sake CPA-1

The use of biocontrol agents (BCA) for controlling plant diseases is an alternative to reduce the use of pesticides. Their performance can be improved when applied in combination with coatings. Films and coatings formulated from different biopolymers were characterized as to their barrier and optical properties to analyse their impact on fruit when applied as carriers of the BCA Candida sake CPA-1. The properties of the film-forming dispersions were more affected by the type of polymer than by the incorporation of surfactants. Sodium caseinate formed the thickest coatings, but these were very thin in every case, which led to there being no predicted relevant effect on the gas exchanges of the fruit. The cell viability in the films was good during film drying, especially in the case of protein films; however, it decreased after storage.

Effect of different coating-forming agents on the efficacy of the biocontrol agent Candida sake CPA-1 for control of Botrytis cinerea on grapes

Multiple formulations of known biocontrol agent (BCA) Candida sake, containing different coating-forming polymers and surfactants were tested at different polymer:BCA ratios, in order to improve control of Botrytis cinerea on grapes. The BCA cell viability on the grape surface was analyzed and reduction in disease incidence and severity was determined. Coating-forming solids improved the survival and efficacy of C. sake as a BCA against B. cinerea, depending on the polymer type and ratio. The incorporation of surfactants did not improve survival or disease control, although they promoted a better cell dispersion on the grape surface. Cell growth of the antagonist during incubation led to the formation of aggregates, even when surfactants were present. Sodium caseinate and starch were the most suitable polymers to formulate C. sake preparations to obtain coating-forming systems with this BCA and to increase its survival and efficacy at the minimum economic cost of the ingredients.

Survival of the biological control agent Candida sake CPA-1 on grapes under the influence of abiotic factors.

As reliability of preharvest applications of biological control agents (BCAs) to control fruit pathogens is highly dependent on the survival of the selected organism, this study aimed to describe the population dynamics of the yeast-BCA Candida sake (Saito & Ota) CPA-1 on grape berries under the effect of abiotic factors such as temperature, relative humidity, sunlight and rainfall. Candida sake (5 × 10(7) CFU ml(-1)), combined with different concentrations of the food additive Fungicover(®), was applied on grape berry clusters. Treated clusters were then exposed to abiotic factors in field or laboratory conditions, recovering populations to describe C. sake population dynamics. The addition of Fungicover significantly increased C. sake multiplication under optimal growth conditions and improved survival under fluctuating abiotic factors. After field applications, significant differences in populations on grape bunches exposed or covered by fine foliage were detected. Simulated rainfall washed off C. sake populations by 0·6-0·9 log units after 20 mm of rain volume. Allowing populations to establish for 24 h or more, prior to a rain event, persistence on grape berries significantly increased and the effect of rain intensity was not observable. Candida sake demonstrated high survival ability under unfavourable environmental conditions and persistence under intense rain. The study evidenced the importance of the first period just after application for C. sake survival on grape tissues and also the protective effect of the additive Fungicover. This research provides new information on the survival of C. sake under field conditions and its practical implications for recommending timing of spray with this antagonist. Our results could be useful for other yeast antagonists applied before harvest. This work, for the first time, defines population dynamics of a yeast BCA using simulated rainfall.

Biological control of Botrytis bunch rot in Atlantic climate vineyards with Candida sake CPA-1 and its survival under limiting conditions of temperature and humidity

Candida sake CPA-1 is an antagonistic yeast that has previously been shown to effectively control Botrytis bunch rot in grapes. The efficacy of biological control agents is dependent on their survival, which may also depend on climatic conditions. However, few studies have evaluated the effect of abiotic factors affecting the survival of biological control agents, such as temperature (T) or relative humidity (RH). In this study, efficacy of C. sake (5×107CFUmL−1), which was applied with the additive Fungicover (FC; 50gL−1), was tested against BBR in the laboratory and in field trials under the Atlantic climate conditions of the Bordeaux region (France). The study also evaluated the survival of C. sake under T and RH regimes simulated in climatic chambers. Two or five applications of C. sake plus FC during the growing season significantly reduced BBR severity at harvest by 48% and 82%, respectively, when compared to the control. Similar reductions were achieved after inoculation with selected virulent Botrytis cinerea strains (75% compared to control) in laboratory experiments. C. sake populations showed minimal decreases between field applications and were favored by simulated Atlantic climate conditions. The survival pattern of C. sake exposed to 40 and 45°C combined with 30% and 100% of RH was described, demonstrating a sharp decrease during the first 24h. Allowing 48h for C. sake to incubate and become established on fruits prior to the exposure to 40°C and 30% RH increased survival (P<0.05). These results confirm the efficacy of treatment with C. sake plus FC under favorable climatic conditions for BBR development, while survival studies may help to improve the survival and efficacy of yeast BCAs, such as C. sake CPA-1.

Fungicide resistance of Botrytis cinerea in tomato greenhouses in the Canary Islands and effectiveness of non-chemical treatments against gray mold.

Tomato greenhouses in the Canary Islands, Spain, were surveyed to estimate frequencies of resistance to benzimidazoles, dicarboximides, anilinopyrimidines and N-phenylcarbamates in Botrytis cinerea. Resistance to carbendazim, iprodione, pyrimethanil and diethofencarb was found in 74.2, 86.4, 28.8 and 31.8% of isolates, respectively. Benzimidazole- and anilinopyrimide-resistant isolates were highly resistant, showing EC50 values above 500 µg/ml carbendazim and a mean EC50 value of 28.42 µg/ml pyrimethanil, respectively. By contrast, a low level of resistance was observed among dicarboximide-resistant isolates (mean EC50 value of 1.81 µg/ml iprodione). Phenotypes with double resistance to carbendazim and iprodione, and triple resistance to carbendazim, iprodione and pyrimethanil were the most common, occurring in 36.4 and 28.8% of isolates. The surveyed greenhouses had never been treated with fenhexamid and Signum™ (pre-packed mixture of boscalid and pyraclostrobin), and baseline sensitivities of B. cinerea isolates to these fungicides were determined. The EC50 values were within the range of 0.009-0.795 µg/ml fenhexamid and of 0.014-0.48 µg/ml Signum. In addition, available formulations based on elicitors of plant defense response and biocontrol agents were evaluated against B. cinerea in tomato plants under semi-controlled greenhouse conditions, the yeast Candida sake CPA-1 being able to reduce gray mold significantly when it was applied on petiole wounds and the plants were inoculated 24 h later. Likewise, C. sake was effective against B. cinerea in harvested tomato fruits, yeast-treated tomatoes showed a 70.66 and 30.31% reduction in the diameters of decay lesions compared with controls after 10 days of storage at 20 and 9 °C, respectively.

Candida sake CPA-1 and other biologically based products as potential control strategies to reduce sour rot of grapes

Sour rot of grapes is becoming increasingly important disease in many wine-growing regions, while consistent chemical or biological control has not been reported. Authors evaluated relative incidence and severity of sour rot in untreated grapevines and the effect of different biologically based treatments on sour rot at harvest. Applications of Candida sake CPA-1 plus Fungicover® , Ulocladium oudemansii and chitosan were carried out in an organic vineyard in Lleida area, Spain, during the 2009 and 2010 growing seasons. At harvest, incidence and severity of sour rot were assessed. Significantly higher incidence and severity of sour rot were observed in untreated plots in 2009, when meteorological conditions after veraison were warmer. All treatments including C. sake CPA-1 significantly reduced (P < 0·05) severity of sour rot in both seasons, ranging from 40 to 67% compared with the untreated control. Incidence of sour rot was not significantly reduced by any treatment. This study helps to characterize development of sour rot in the dry Mediterranean climate conditions of the experiment, whereas also represents the first report of biological control of sour rot. Treatments with the tested biologically based products are a promising strategy to control sour rot. Studies on sour rot of grapes are scarce in literature, and this is the first work specifically evaluating sour rot in Spanish vineyards. Sour rot control in field conditions through applications of antagonistic micro-organisms is reported for first time in this study, showing elevated severity reductions (40-67% compared with control). As there are no options available for sour rot control in vineyards, results point Candida sake CPA-1 as an interesting control strategy against grape bunch rots.

Field applications of improved formulations of Candida sake CPA-1 for control of Botrytis cinerea in grapes

There is increasing interest in the use of biological control agents (BCAs) to replace synthetic fungicides. The aim of this study was to evaluate the effectiveness of combining different formulation strategies on the establishment, survival, persistence and efficacy of Candida sake CPA-1 applied in the field in order to control Botrytis cinerea in grapes. Different formulations of C. sake cells were applied at flowering, pea sized berries, veraison and before harvest. In 2005, heat adapted or non-adapted fresh cells of C. sake were tested. In 2006, these same treatments were tested but, in this case, the C. sake cells were also formulated in an isotonic solution. The compound Fungicover® (FC) an edible coating was evaluated as a potential additive for C. sake treatments. Spray applications of different formulations of C. sake resulted in colonisation of bunches under field conditions and when combined with FC, C. sake had significantly higher survival rates (up to 50% higher) compared with C. sake without additive FC. Formulation of C. sake cells in an isotonic solution combined with FC resulted in Botrytis spp. control similarly than fungicide treatment. The potential of C. sake for biocontrol of Botrytis bunch rot of grapevine has been demonstrated. FC has a beneficial effect on the BCA, improving the persistence of C. sake cells on the host and its efficacy to levels comparable to fungicide treatment. It is possible to broaden the spectrum of use of BCAs using different formulation strategies and to thereby develop practical uses under field conditions.

Impact of mild heat treatments on induction of thermotolerance in the biocontrol yeast Candida sake CPA-1 and viability after spray-drying

The objective of this study was to examine the induction of thermotolerance in the biocontrol agent Candida sake CPA-1 cells by mild heat treatments to enhanced survival of formulations using spray-drying. The possible role of heat-shock proteins (HSPs) biosynthesis in induced thermotolerance and the role of sugars and sugar alcohols were also determined. Studies were conducted on C. sake cells grown in molasses medium and exposed to mild temperatures of 30 and 33 degrees C during mid- (16 h), late-exponential (24 h), early- (30 h) and mid-stationary (36 h) growth phases. The effect on viability was determined both before and after spray-drying. Cycloheximide and chloramphenicol were used to examine the role of HSPs and HPLC was used to analyse the accumulation of sugar and sugar alcohols. The results indicate that both temperatures induced thermotolerance in cells of C. sake. Mild heat-adapted cells at 33 degrees C in the early- or mid-stationary phases had survival values after spray-drying significantly higher (P <or= 0.05) than nonadapted cells. However, viabilities were not high enough to be considered for commercial use with values up to 17%. HSPs were not implicated in thermotolerance acquired by mild heat-adapted cells as similar viabilities were obtained in the presence of protein inhibitors. Little change was observed in sugar and sugar alcohols with an increase in glucose and arabitol in some treatments. This study suggests that it is possible to induce thermotolerance in biocontrol yeasts such as C. sake. However, this does not improve survival of cells exposed to spray-drying sufficiently to consider this a suitable formulation method for this biocontrol agent. HSPs, sugars and sugar polyols were not directly responsible for induced thermotolerance in yeast cells. This type of information can be effectively applied to improve the viability of cells in the process of formulation.

Efficacy of Candida sake CPA-1 Formulation for Controlling Penicillium expansum Decay on Pome Fruit from Different Mediterranean Regions

The effectiveness of a formulated product of the yeast Candida sake CPA-1 for controlling postharvest diseases on pome fruits was demonstrated in laboratory, semicommercial, and commercial trials carried out in the major pome fruit producing region of the European Union. First, one wettable powder and seven liquid formulations were tested in laboratory trials that involved two varieties of apples and two varieties of pears. In all cases, an efficacy similar to that of fresh cells was demonstrated in the control of artificial Penicillium expansum infection. After these trials, the formulated product chosen for semicommercial and commercial trials was LF1, a liquid formulation that is particularly suitable for commercial applications. In semicommercial trials, LF1 showed a performance similar to fresh cells in most trials, and the population dynamics of both fresh and formulated cells were quite stable throughout the storage period. This indicates the high viability of C. sake CPA-1 in this formulation and the absence of adverse effects during the formulation of the product, which may significantly affect both its ability to grow on fruit and its antagonistic activity. We evaluated the control of natural infection after applying the formulated product in a commercial drencher in different packinghouses. A significant reduction in the incidence of diseases was observed with a recommended dose of around 107 CFU/ml when natural infections were greater than 1%. In general, large quantities of yeast were observed on the surface of unwounded fruits of different pome fruit cultivars. Moreover, populations of this biocontrol agent increased rapidly on fruit surfaces and remained quite stable for a long time under commercial storage conditions. Commercial practices used in packinghouses were therefore successfully applied for this formulated product.

Survival of the postharvest biocontrol yeast Candida sake CPA-1 after dehydration by spray-drying

Spray drying was evaluated as a dehydrating method to preserve the postharvest biocontrol agent Candida sake CPA-1. The effect of drying temperature, carrier, growth and rehydrating medium on the survival of the yeast was studied. Outlet temperature had more influence on the death of the cells than inlet temperature, and survival decreased with increasing temperature. Spray drying at an inlet temperature of 150°C was optimum in terms of viability, powder recovery and moisture content of the product. Use of 10% (v/v) skimmed milk as a carrier gave the highest survival and percentage of powder recovery (34–47%). Rich rehydration media were found to be better than water or phosphate buffer, with slight differences on survival. Spray-dried cells were less effective than fresh ones in controlling Penicillium expansum rot on apples. Spray drying of C. sake was not a good dehydration method as it gave low cell survival, poor recovery of product, and low efficacy.

BIOCONTROL OF POSTHARVEST BLUE MOULD ON PEAR AND APPLE FRUITS WITH THE COMBINATION OF CANDIDA SAKE (CPA-1) AND PSEUDOMONAS SYRINGAE (CPA-5)

The effectiveness of Candida sake (CPA-1) in combination with Pseudomonas syringae (CPA-5) for controlling P. expansum on pears and apples was determined. The concentrations tested were 5×10 6 cfu/ml for C. sake and 2×10 7 cfu/ml for P. syringae. At room temperature the two antagonists were combined in proportions of 0 to 100% in 25% increments. The best combinations were chosen for the assays under cold temperature and different atmosphere conditions. Population dynamics of C. sake (CPA-1) and P. syringae (CPA-5) in mixture or individual application in wounded fruits were determined and showed the same growth pattern when applied alone or in combination. From the experimental results it can be concluded that the combination of C. sake CPA-1 and P. syringae CPA-5 improves the control of P. expansum in apple and pears fruits and allows the requeired concentration to achieve control to be reduced, with the consequent reduction in the cost of the treatment.

Optimization of growth conditions of the postharvest biocontrol agent Candida sake CPA-1 in a lab-scale fermenter.

To maximize the growth (expressed as number of viable cells per millilitre) of the postharvest biocontrol agent Candida sake CPA(-1) at laboratory scale conditions. Growth conditons (aeration, agitation speed and inoculum size) were studied in batch conditions in a 5 l fermenter using molasses and urea as growth medium. Consumption of sugars and urea were analysed. Fed-batch studies were also carried out. Glucose and fructose were consumed during the exponential growth phase and were depleted after 18 h of growth. On the contrary, C. sake cells assimilated sucrose during the stationary phase. There was not growth improvement when fed-batch technology was used. Addition of an extra amount of glucose or molasses after 18 h of growth did not contribute to increase final population. Maximum growth (about 8 x 10(8 )CFU ml(-1)) was obtained at batch fermentation after 30 h growth at 400 rev min(-1), 150 l h(-1) of air and initial concentration of 106 CFU ml(-1). The results obtained in this study are an approach for further upscaling of C. sake production.