Progress 08/01/17 to 07/31/22
Outputs
Target Audience:Due to support obtained through this project, the PI has been able to train a graduate student in basic microbiology, controlled environment agriculture and produce safety. In addition, results of the study have been shared with the larger community including, researchers, academics, regulators and industry partners. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Through this project, a graduate and undergraduate student were trained in conducting research in microbiology and produce safety. Specifically, the students were engaged in conducting the experiments evaluating the antimicrobial potential of probiotics and biocides when applied on seeds and sprouts. In addition to the wet lab experiments, the graduate student also performed statistical analysis on the data and is being trained in scientific writing. How have the results been disseminated to communities of interest?Results of the study were shared with the scientific community, food safety professionals, industry experts and regulators through presentations at regional, national and international meetings. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Obj. 1: Since contaminated seeds is the primary reason for contaminated sprouts, controlling pathogens on seeds is the first step to ensure sprouts safety. Hence in objective 1, experiments were set up to evaluate the antimicrobial efficacy of lactic cultures in combination with peroxyacetic acid (PAA) wash in reducing Salmonella (SE) and E. coli O157 (EC) populations on alfalfa seeds over a 12-month period. Briefly, alfalfa seeds were surface sterilized and inoculated with Salmonella or EHEC cocktail (~5 log CFU/g). Following inoculation and drying, the seed were rinsed in 80 ppm PAA, dried and sprayed with different probiotics (Lactococcus lactis B-23802, L. lactis B-23804, Lactobacillus acidophilus LA5 and Hafnia alvei B16; ~9 log CFU/g). The seeds were then dried, transferred to sterile stomacher bags and stored under ambient conditions until sampling. Samples were analyzed at regular intervals through one year of storage. Multi-hurdle approach using probiotics and organic acids significantly reduced EC populations on seeds and sprouts (p≤0.05). By one month of storage, treatment with PAA and probiotics reduced EC and SE population on seeds to below detection limits. However, ~2-3 log CFU of the pathogen was recovered from the control seeds. Further, PAA and probiotic- treated samples were found to be enrichment negative for the pathogen (SE/EC) by 6-9 months of storage while control samples were enrichment positive until the end of the 12-month storage period. Also, ~4-6 log CFU/g of the lactic cultures was recovered from the seeds at the end of the study. These results demonstrate that probiotic strains in combination with PAA could serve as a viable seed treatment to reduce pathogen contamination on sprout seeds. Obj. 2: Besides contaminated incoming material (seeds), risk-based analysis has identified several stages during sprout production as being critical to control foodborne pathogens. These stages include sprouting and sprout growth. The warm and humid sprouting process provides an ideal environment for pathogens to grow, thus a low level of contamination by Salmonella and E. coli O157 on the seeds could eventually get to high pathogen populations. Therefore, as means to control pathogen growth during sprouting and reduce overall contamination, we employed a multi hurdle approach incorporating a PAA rinse, probiotic pre-germination soak and a lactic acid post-harvest rinse. Pre-harvest: Overall, pathogen populations in the control increased over the germination period (2.5 log to 9 log at the end of 7 days). Results of these experiments revealed that the combinatorial effect of pre-rinse with PAA, pre-germination soak with probiotic treatments and a post-harvest rinse with lactic acid significantly (p≤0.05) reduced pathogen populations on the sprouts when compared to the untreated control. Following the PAA rinse and probiotic soak, on day 1 of germination, approximately 4 and 6 log CFU of the pathogen was recovered from the treatment and control seeds, respectively. By day seven of germination, approximately 9 and 7.2 log CFU of the pathogen was recovered from the treated and control samples. Post-harvest: To further reduce pathogen populations on the germinated sprouts, 2% lactic acid was evaluated as a potential post-harvest wash. While PAA and probiotic treatment led to a 2-3 log reduction in pathogen populations, incorporation of a lactic acid post-harvest rinse led to ~ 4 log reduction in pathogen populations when compared to the control (~8-9 log CFU/g). Additionally, approximately 5-6 log CFU of LAB were recovered from the seeds and sprouts by the end of the study. Hence use of probiotics and biocides in a multi-hurdle approach was found to significantly control pathogen populations on sprouts. Spent water: Since spent irrigation water can serve as a source of pathogen contamination for sprouts, triplicate water samples were collected at each sampling time during germination for microbiological analysis. In addition to reducing pathogen populations on the sprouts, PAA and LAB treatments significantly reduced pathogen counts in spent water. For instance, with the control samples, approximately 8 log CFU/ ml of EC was recovered from the spent water throughout the 7-day study period. Application of a PAA seed rinse and pre-germination soak containing LAB reduced pathogen load in the irrigation water by greater than 2 logs by the end of the study. Seed germination and sprout growth: With regards to sprout germination and quality, overall, the germination percentage ranged from 87-96%. Overall, PAA pre-rinse followed by probiotic application did not result in a significant reduction in germination when compared to the control (96%). Further, these treatments did not affect sprout growth. On day 5, germinated sprouts were assessed for growth by measuring sprout length. Based on established standards, the typical sprout length for alfalfa sprouts is expected to be 26-38 mm. In our study, all treatments had a sprout length similar to the control with an average sprout length of 41.22 mm. Summary and project outcomes: Overall, Application of biocides in combination with probiotics significantly reduce pathogen contamination on seeds without impacting seed germination and sprout growth. Further, use of probiotics in combination with PAA and LA in a multi-hurdle approach reduced pathogen contamination on sprouts. Therefore, this multi-hurdle approach could be used as an effective, organic-friendly approach to improve sprout safety.
Publications
- Type:
Other
Status:
Other
Year Published:
2022
Citation:
Research presentation on " A natural, multi-hurdle approach to control�Salmonella�and E. coli O157:H7 on alfalfa sprouts" at the IAFP 2022 3 MT competition
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Progress 08/01/20 to 07/31/21
Outputs
Target Audience:The target audience includes the scientific community, food microbiologists, produce industry and graduate and undergraduate students at UConn. Changes/Problems:The impact of COVID and research ramp down led to a significant delay in the study. Specifically, inability of research personnel to come to campus delayed research progress. We were unable to set up experiments and lost several samples from previous trails that were started before the shutdown. Particularly with the yearlong seed experiments, samples are collected and analyzed regularly through one year of storage. For experiments set up in Jan/Feb 2020, we missed sampling times in March, April, May, June, July, Aug. We have missing data for 6 months. Also, these experiments no longer correspond to the previous trials that were conducted in 2018 and 2019. So, we had to discard that experiment and set up new trials. One set of trials were set up in October 2020 and will run till Oct 2021. The next set of trials were set up in Feb 2021 and will run until Feb 2022. What opportunities for training and professional development has the project provided?During this reporting period, a graduate and undergraduate student were trained in conducting research in microbiology. Specifically, the students were engaged in conducting the experiments evaluating the antimicrobial potential of probiotics when applied on seeds and sprouts. In addition to the wet lab experiments, the graduate student also performed statistical analysis on the data. How have the results been disseminated to communities of interest?Results of the study will be presented at the upcoming IFT and IAFP 2021 annual meeting this summer. What do you plan to do during the next reporting period to accomplish the goals?The impact of COVID and research ramp down led to a significant delay in the study. Specifically, inability of research personnel to come to campus delayed research progress. We were unable to set up experiments and lost several samples from previous trails that were started before the shutdown. Particularly with the yearlong seed experiments, samples are collected and analyzed regularly through one year of storage. For experiments set up in Jan/Feb 2020, we missed sampling times in March, April, May, June, July, Aug. We have missing data for 6 months. Also, these experiments no longer correspond to the previous trials that were conducted in 2018 and 2019. So, we had to discard that experiment and set up new trials. One set of trials were set up in October 2020 and will run till Oct 2021. The next set of trials were set up in Feb 2021 and will run until Feb 2022. Further, additional trials will be set up to replicate and validate results obtained with pathogen control on sprouts and wash water and sprout germination and quality assays.
Impacts
What was accomplished under these goals?
During the past reporting period, experiments were conducted to evaluate the antimicrobial effect of different probiotic strains and biocides in controlling Salmonella and E. coli O157 on alfalfa seeds and sprouts. Under objective 1, experiments were set up to evaluate the antimicrobial efficacy of lactic cultures in reducing Salmonella (SE) and E. coli O157 (EC) populations on alfalfa seeds over a 12-month period. Briefly, alfalfa seeds were surface sterilized and inoculated with Salmonella or EHEC cocktail (~5 log CFU/g). Following inoculation and drying, the seed were rinsed in 80 ppm PAA, dried and sprayed with different probiotics (Lactococcus lactis B-23802, L. lactis B-23804, Lactobacillus acidophilus LA5 and Hafnia alvei B16; ~9 log CFU/g). The seeds were then dried, transferred to sterile stomacher bags and stored under ambient conditions until sampling. Samples were analyzed at regular intervals through one year of storage. Multi-hurdle approach using probiotics and organic acids significantly reduced EC populations on seeds and sprouts (p≤0.05). By one month of storage, treatment with PAA and probiotics reduced EC and SE population on seeds to below detection limits. However, ~2-3 log CFU of the pathogen was recovered from the control seeds. Also, ~4-6 log CFU/g of the lactic cultures was recovered from the seeds at the end of six months. Under objective 2, experiments were conducted to evaluate the antimicrobial efficacy of probiotic cultures in combination with PAA and lactic acid to control pathogens on growing sprouts. Alfalfa seeds were surface sterilized and inoculated with Salmonella or EHEC cocktail (~3 log CFU/g) and treated with 80 ppm PAA for 10 min as a pre-germination seed rinse. The seeds were then treated with different probiotic strains (~ 9 log CFU/g) as a pre-germination soak for 6 h. The seeds were then set to sprout in the growth chamber. At each sampling time, day 0 (prior to germination), 1, 3, and 5 of germination, triplicate 10 g samples were processed, and surviving Salmonella/EHEC and LAB populations were enumerated. Additionally, on day 5, a sub-set of the samples were rinsed in 2% lactic acid as a post-harvest wash. Sprouts were sampled immediately after the post-harvest wash. Control samples were rinsed in sterile water and sampled to assay surviving pathogen population. Overall, pathogen populations in the control increased over the germination period (6.5 log to 9 log at the end of 5 days). Results of these experiments revealed that the combinatorial effect of pre-rinse with PAA, pre-germination soak with probiotic treatments and a post-harvest rinse with lactic acid significantly (p≤0.05) reduced pathogen populations on the sprouts when compared to the untreated control. Following the PAA rinse and probiotic soak, on day 1 of germination, approximately 3 and 6 log CFU of the pathogen was recovered from the treatment and control seeds, respectively. By day five of germination, approximately 9 and 6 log CFU of the pathogen was recovered from the treated and control samples. To further reduce pathogen populations on the germinated sprouts, 2% lactic acid was evaluated as a potential post-harvest wash. While PAA and probiotic treatment led to a 2-3 log reduction in pathogen populations, incorporation of a lactic acid post-harvest rinse led to ~ 4 log reduction in pathogen populations when compared to the control (~8-9 log CFU/g). Additionally, approximately 5-6 log CFU of LAB were recovered from the seeds and sprouts by the end of the study. Hence use of probiotics and biocides in a multi-hurdle approach was found to significantly control pathogen populations on sprouts. With regards to sprout germination and quality, overall, the germination percentage ranged from 87-96%. PAA pre-rinse followed by probiotic application led a reduction in germination when compared to the control (96%). However, these treatments did not affect sprout growth. On day 5, germinated sprouts were assessed for growth by measuring sprout length. Based on established standards, the typical sprout length for alfalfa sprouts is 26-38 mm. In our study, all treatments had a sprout length similar to the control with an average sprout length of 41.22 mm.
Publications
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Progress 08/01/19 to 07/31/20
Outputs
Target Audience:The target audience this reporting period included the scientific community interested in the study of foodborne illness and microbiological safety of produce, and students at UConn. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?During this reporting period, a graduate student was trained in conducting research in microbiology. Specifically, the student was engaged in conducting the experiments evaluating the antimicrobial potential of probiotics when applied on seeds and sprouts. In addition to the wet lab experiments, the graduate student also performed statistical analysis on the data. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?In continuation of the proposed research activities, additional experiments will be set up to evaluate the efficacy of probiotic application on pathogen viability/survival on sprouts seeds and germinating sprouts as described under Objective 1 and 2. Additionally, storage experiments will be performed to investigate the potential application of probiotics and biocides as a pre-germination wash and its efficacy as a post-harvest antimicrobial hurdle on sprouts during storage.
Impacts
What was accomplished under these goals?
During the past reporting period, experiments were conducted to evaluate the antimicrobial effect of different probiotic strains and biocides in controlling Salmonella and E. coli O157 on alfalfa seeds and sprouts. Under objective 1, experiments were set up to evaluate the antimicrobial efficacy of lactic cultures in reducing Salmonella and E. coli O157 populations on alfalfa seeds over a 12-month period. Briefly, alfalfa seeds were surface sterilized and inoculated with Salmonella or EHEC cocktail (~6.5 log CFU/g) and treated with different probiotic strains (LA/ LL/SLD/LR/LP; ~8 log CFU/g). The seeds were then transferred to sterile stomacher bags and stored at room temperature until sampling. The results indicate that Salmonella was significantly more adept at surviving on the seeds than EHEC. For example, 4 log CFU of Salmonella was recovered from the seeds following nine months of storage. On the other hand, EHEC populations were reduced to less than 3 log by six months of storage and undetectable by 9 months. In the probiotic-treated samples, all probiotics reduced Salmonella populations to below detection limits by end of the study. As with the control samples, the probiotics were significantly more effective in controlling EHEC on seeds when compared to Salmonella. For instance, probiotics (LA/LL/LP/LR) reduced EHEC to below detection limits by 6 months of storage while this reduction was observed only by 12 months with Salmonella. Also, ~4 log CFU/g of the lactic cultures was recovered from the seeds at the end of twelve months. Under objective 2, experiments were conducted to evaluate the antimicrobial efficacy of probiotic cultures in combination with PAA to control pathogens on growing sprouts. Alfalfa seeds were surface sterilized and inoculated with Salmonella or EHEC cocktail (~3 log CFU/g) and treated with 80 ppm PAA for 10 min as a pre-germination seed rinse. The seeds were then treated with different probiotic strains (HA/LL/LA/BB; ~ 7.5 log CFU/g) as a pre-germination soak for 6 h. The seeds were then set to sprout in the growth chamber. At each sampling time, day 0 (prior to germination), 1, 3, and 5 of germination, triplicate 10 g samples were processed, and surviving Salmonella/EHEC and LAB populations were enumerated. Additionally, on day 5, a sub-set of the samples were rinsed in either 80 ppm PAA or 2% lactic acid as a post-harvest wash and subjected to refrigerated storage for 5 days. Sprouts were samples immediately after the post-harvest wash and following refrigerated storage. Overall, pathogen populations in the control increased over the germination period (6.5 log to 9 log at the end of 5 days). Results of these experiments revealed that the combinatorial effect of pre-rinse with PAA followed by probiotic treatments significantly reduced initial pathogen populations on the seeds when compared to the untreated control. For instance, on day 1 of germination, approximately 3 and 6 log CFU of the pathogen was recovered from the treatment and control seeds, respectively. By day five of germination, approximately 9 and 6 log CFU of the pathogen was recovered from the treated and control samples. To further reduce pathogen populations on the germinated sprouts, 80 ppm PAA and 2% lactic acid were evaluated as a potential post-harvest wash. Initial trials indicate that lactic acid is more effective in reducing pathogen populations on sprouts when compared to PAA. Washing sprouts in 2% lactic acid reduced Salmonella populations to less than 3.5 log CFU/g when compared to sprouts washed in water or PAA. Further, no significant increase in pathogen population was observed in sprouts following refrigerated storage. Similar results were observed in the EHEC trials. Hence use of probiotics and biocides in a multi-hurdle approach was found to significantly control pathogen populations on sprouts. Since spent irrigation water can serve as a source of pathogen contamination for sprouts, triplicate water samples (10 ml each) were collected at each sampling time during germination for microbiological analysis. PAA and LAB treatments significantly reduced pathogen counts in spent water. With the control samples, approximately 7.5 log CFU/ ml of Salmonella/EHEC was recovered from the spent water throughout the 5-day study period. Application of a PAA seed rinse and pre-germination soak containing LAB reduced pathogen load in the irrigation water by greater than 5 logs by the end of the study. In addition to enumerating pathogen populations, sprouts and seeds were sampled to evaluate LAB survival. Consistently throughout the study, approximately 7 log CFU of LAB cultures were recovered from growing sprouts and spent irrigation water.
Publications
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Progress 08/01/18 to 07/31/19
Outputs
Target Audience:Results of this study were presented to the scientific community and students at UConn and New England region through presentations at meetings Changes/Problems:Use of dip inoculation to test the antimicrobial efficacy of probiotics on seeds was associated with fungal contamination during seed storage. Therefore, spray application ofpathogen and probiotics will be evaluated as a means to minimize/eliminate fungal contamination in seed samples. Further, the antimicrobial efficacy of the LAB strains used against a pathogen cocktail was found to be reduced when compared to our preliminary study using a single strain of the pathogen. Hence, additional lactic cultures have been obtained from USDA ARS to evaluate their antimicrobial efficacy. We will test these strains against the pathogen cocktail to identify effective LAB treatments. What opportunities for training and professional development has the project provided?During this reporting period, a graduate and undergraduate student were trained in conducting research in microbiology. Specifically, the students were engaged in conducting the experiments evaluating the antimicrobial potential of probiotics when applied on seeds and sprouts. In addition to the wet lab experiments, the graduate student also performed statistical analysis on the data. How have the results been disseminated to communities of interest?Results of the study have been disseminated to the scientific community through poster presentations. What do you plan to do during the next reporting period to accomplish the goals?In continuation of the proposed research activities, additional experiments are currently underway to evaluate the efficacy of probiotic application on pathogen viability/survival on sprouts seeds and germinating sprouts as described under Objective 1 and 2.
Impacts
What was accomplished under these goals?
During the past reporting period, experiments were conducted in the lab to standardize the inoculation procedure to artificially contaminate sprout seeds and for probiotic inoculation. Further, an MTA was executed between UConn and USDA-ARS Beltsville to obtain produce outbreak associated O157 STEC cultures for use in the study. Further, additional lactic cultures were obtained from USDA ARS culture collection to evaluate their antimicrobial efficacy. Under objective 1, Year long experiments were set up to evaluate the antimicrobial efficacy of lactic cultures in reducing Salmonella and E. coli O157 populations on alfalfa seeds. Briefly, alfalfa seeds were surface sterilized and inoculated with Salmonella or EHEC cocktail (~6.5 log CFU/g) and treated with different probiotic strains (LA/ SLD/LR; ~6.5 log CFU/g). The seeds were then transferred to sterile stomacher bags and stored at room temperature until sampling. These initial trials have demonstrated that Salmonella and EHEC are adept at surviving on seeds in significant numbers. At the end of nine months, approximately 5 log CFU/g of the pathogen was recovered from the stored seeds. On the contrary, treatment with lactic cultures (LP, LL, LA and SLD) resulted in a 0.5- 2 log reduction in pathogen populations. Also, ~7 log CFU/g of the lactic cultures was recovered from the seeds at the end of nine months. Under objective 2, experiments were conducted to evaluate the antimicrobial efficacy of lactic cultures to control pathogens on growing sprouts. Alfalfa seeds were surface sterilized and inoculated with Salmonella cocktail (~6.5 log CFU/g) and treated with different probiotic strains (LA/ LL/SLD/LR; ~6.5 log CFU/g). The seed were then set to sprout in the growth chamber. At each sampling time day 0 (prior to germination), 1, 3, 5, 7 and 9 of germination, triplicate 10 g samples were processed and surviving Salmonella and LAB populations were enumerated. Overall, Salmonella populations in the control increased over the germination period (6.5 log to 8.6 log at the end of 9 days). Treatment with different LAB strains reduced Salmonella populations throughout the growing period by 1-4 logs by the end of the study. Since spent irrigation water can serve as source of pathogen contamination for sprouts, triplicate water samples (10 ml each) were collected at each sampling time for microbiological analysis. Similar to Salmonella reduction on sprouts, LAB treatments significantly reduced pathogen counts in spent water. With the control samples, approximately 7.5 log CFU/ ml of Salmonella was recovered from the spent water throughout the nine-day study period. Application of a pre-germination wash containing LAB, reduced pathogen load in the irrigation water by 2-4 logs by the end of the study. In addition to enumerating pathogen populations, sprouts and seeds were sampled to evaluate LAB survival. Consistently throughout the study, approximately 7 log CFU of LAB cultures were recovered from growing sprouts and spent irrigation water.
Publications
- Type:
Other
Status:
Published
Year Published:
2019
Citation:
Poster presentation at the UConn College of Agriculture, Health and Natural Resources Graduate Student Forum, "Controlling Salmonella on sprouts using lactic cultures"
- Type:
Other
Status:
Published
Year Published:
2019
Citation:
Poster Presentation at Pioneer Valley Microbiology Symposium
"Controlling Salmonella on alfalfa sprouts using lactic cultures"
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Progress 08/01/17 to 07/31/18
Outputs
Target Audience:
Nothing Reported
Changes/Problems:With objective 1, use of dip inoculation to test the antimicrobial efficacy of probiotics on seeds was associated with fungal contamination during seed storage. Therefore, spray application ofpathogen and probiotics will be evaluated as a means to minimize/eliminate fungal contamination in seed samples. What opportunities for training and professional development has the project provided?During this reporting period, a graduate and undergraduate student were trained in conducting research in microbiology. Specifically, the students were engaged in conducting the experiments evaluating the antimicrobial potential of probiotics when applied on seeds and sprouts. In addition to the wet lab experiments, the graduate student also performed statistical analysis on the data. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?In continuation of the proposed research activities, additional experiments will be set up to evaluate the efficacy of probiotic application on pathogen viability/survival on sprouts seeds and germinating sprouts as described under Objective 1 and 2. Additionally, storage experiments will be performed to investigate the potential application of probiotics as a pre-germination wash and its efficacy as a post-harvest antimicrobial hurdle on sprouts during storage.
Impacts
What was accomplished under these goals?
During the past reporting period, experiments were conducted in the lab to standardize the inoculation procedure to artificially contaminate sprout seeds and for probiotic inoculation. Further, an MTA was executed between UConn and USDA-ARS Beltsville to obtain produce outbreak associated O157 STEC cultures for use in the study. Efficacy of probiotics in reducing Salmonella/non-O157 STEC populations on germinating sprouts: Alfalfa seeds were surface sterilized and inoculated with Salmonella /non-O157 STEC cocktail (~6.5 log CFU/g) and treated with different probiotic strains (LA/ SLD/LR; ~6.5 log CFU/g) as described under objective 1. The seed were then set to sprout in the growth chamber. At each sampling time day 0 (prior to germination), 1, 3, 5, 7 and 9 of germination, triplicate 25 g samples were processed and surviving Salmonella/non-O157 STEC and probiotic population were enumerated. Application of different probiotics significantly reduced pathogen populations on germinating sprouts. Treatment with LA, LR and SLD significantly reduced Salmonella/non-O157 STEC populations. Specifically, treatment with LR and SLD reduced pathogen populations to below undetectable levels by day 3 of sprouting. Application of LA reduced pathogen populations on sprouts by ~ 2 and 3.5 log by day 1 and 3 of sprouting. However ~8 log CFU/g of Salmonella/non-O157 STEC was still recovered from the control sprouts at the end of the experiment. Efficacy of probiotics application on Salmonella/non-O157 STEC populations in spent irrigation water: Water samples (three-25 ml samples) from the bottom chamber was collected at regular intervals (day 1, 3, 5, 7, 9) for microbiological analysis. As seen with the germinating sprouts, probiotic application significantly reduced pathogen population in the spent water when compared to the control. Approximately 4 - 6 log CFU of Salmonella/non-O157 STEC was recovered from the water samples throughout the study. However, treatment with LR and SLD reduced pathogen populations to below undetectable levels by day 1of sprouting. With LA, water samples were negative for Salmonella/non-O157 STEC by day 5 of the experiment. Effect of probiotic supplementation on seed viability and germination: Seeds treated with the different probiotic strains and control seeds (untreated) were set up for germination and sprouting in the germination chamber. These seeds were observed for visibly protruding radical and the number of germinated seeds was counted after 24 and 72 h. It was observed that probiotics did not have any detrimental effect on seed viability and germination. Moreover application of probiotics significantly improved germination. Specifically, 24 h after experimental set up, germination ranged from 56-58% for probiotic treated seed when compared to 42% in the control. Similarly, following 72 h, germination was around 87% and 69% in the probiotic and control groups, respectively.
Publications
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