Progress 10/01/12 to 09/30/13
Outputs
Progress Report Objectives (from AD-416): 1. Quantify the compositional and microbial changes of noni juice during the fermentation period. 2. Develop a standard method for processing fermented noni juice. Approach (from AD-416): Physiologically mature, ripe noni fruit will be harvested from commercial orchards on the island of Hawaii. Fruit will be washed and placed into sterilized glass jars with sealed lids and allowed to age for 10-14 days at 25�C. A second batch will be aged for up to 30 days. Juice will be pasteurized at 85�C in a water bath, cooled, and frozen. Subsamples of juice will be collected throughout the fermentation process to conduct compositional and microbial analyses. Prominent yeast, bacteria, and fungi will be isolated and identified using standard microbiological techniques. Ascorbic acid, malic acid, lactic acid, and sugar concentrations will be analyzed using high performance liquid chromatography (HPLC). Total antioxidant activity will be determined using a photochemical luminescence method. Collaborators at the University of Hawaii, College of Pharmacy will test samples of noni juice for bioactivity and toxicity in various cell culture assays. Documents Trust with Patient First C Foundation. Log 39444. Formerly 5320-43000-015- 08T (06/11). The goal of this project is to quantify the compositional and microbial changes of noni juice during the fermentation period and to develop a standard method for processing fermented noni juice which relates to the overall objective of the parent project to develop and protect U.S. export markets for fresh tropical commodities with emphasis on expanding and diversifying agriculture and agricultural exports in Hawaii and other states by providing environmentally sound, economically viable systems, treatments, or processes that control quarantine pests, ensure product quality and food safety, and increase product value while safeguarding the agriculture of other states. Noni fruit (Morinda citrifolia) and juice extracts are reportedly therapeutic for diabetes, high blood pressure, and certain types of cancer. Research was completed to determine the physiology of noni fruit ripening, as well as the chemical and microbial changes that occur during noni juice fermentation. Ripening noni fruit had a nonclimacteric respiratory pattern and no detectable ethylene production. The fungus, Mucor circinelloides f. sp. circinelloides, was consistently isolated from fermented noni juice, with peak populations at 14 days. The bacteria Erwinia pyrifoliae and Gluconobacter frateurii, were isolated from fermented juice at 42 days. Total sugar concentration decreased by 37% after 7 days, whereas total organic acids were highest at 28 days. The major non-volatile acids identified in noni juice included acetic, ascorbic, dehydroascorbic, galacturonic, malonic, succinic, and tartaric acids. Ethanol and acetic acid were the main fermentation products detected in noni juice. The optimum fermentation time was 14 to 21 days, and the need for pasteurization was clearly demonstrated. These results will be useful in developing a standard method for producing consistent quality noni juice.
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Progress 10/01/11 to 09/30/12
Outputs
Progress Report Objectives (from AD-416): 1. Quantify the compositional and microbial changes of noni juice during the fermentation period. 2. Develop a standard method for processing fermented noni juice. Approach (from AD-416): Physiologically mature, ripe noni fruit will be harvested from commercial orchards on the island of Hawaii. Fruit will be washed and placed into sterilized glass jars with sealed lids and allowed to age for 10-14 days at 25�C. A second batch will be aged for up to 30 days. Juice will be pasteurized at 85�C in a water bath, cooled, and frozen. Subsamples of juice will be collected throughout the fermentation process to conduct compositional and microbial analyses. Prominent yeast, bacteria, and fungi will be isolated and identified using standard microbiological techniques. Ascorbic acid, malic acid, lactic acid, and sugar concentrations will be analyzed using high performance liquid chromatography (HPLC). Total antioxidant activity will be determined using a photochemical luminescence method. Collaborators at the University of Hawaii, College of Pharmacy will test samples of noni juice for bioactivity and toxicity in various cell culture assays. The goal of the Trust Agreement is to quantify the compositional and microbial changes of noni juice during the fermentation period and to develop a standard method for processing fermented noni juice. Noni fruit (Morinda citrifolia) are used to produce fermented juice that often varies in chemical, physical, microbial, and medicinal properties. We completed chemical analyses of noni juice sugars, organic acids, and ethanol concentrations over 28 days of fermentation using liquid chromatography. The results reveal fermentation end-points for standardizing the processing of noni juice. Also, the antifungal properties of pasteurized (85 �C, 3 min) noni juice exudate were determined in spot-plate bioassays against several fungal plant pathogens of tropical fruit crops. In bioassays of fresh or fermented noni juice, antifungal activity was absent in juice with 5% soluble solids (TSS). However, fermented noni juice with 7% TSS or higher completely inhibited spore germination of five plant fungal pathogens (Lasmenia sp., Colletotrichum acutatum, C. gloeosporioides, Gliocephalotrichum bulbilium, G. simplex). In related research, large volumes of noni juice were prepared for partitioning with n-butanol and ethyl acetate. Previously, our collaborators reported that fermented noni juice demonstrated antitumor activity, and that the n-butanol and ethyl acetate fractions contained active components. The collected partitions were further fractionated into five n-butanol and eight ethyl acetate subfractions to identify specific subfractions with antitumor activity in bioassays and mice models by collaborators. The work in the Trust Agreement will contribute directly to objective 2 of the parent project to develop new or improved postharvest treatments and enhance product value.
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Progress 10/01/10 to 09/30/11
Outputs
Progress Report Objectives (from AD-416) 1. Quantify the compositional and microbial changes of noni juice during the fermentation period. 2. Develop a standard method for processing fermented noni juice. Approach (from AD-416) Physiologically mature, ripe noni fruit will be harvested from commercial orchards on the island of Hawaii. Fruit will be washed and placed into sterilized glass jars with sealed lids and allowed to age for 10-14 days at 25�C. A second batch will be aged for up to 30 days. Juice will be pasteurized at 85�C in a water bath, cooled, and frozen. Subsamples of juice will be collected throughout the fermentation process to conduct compositional and microbial analyses. Prominent yeast, bacteria, and fungi will be isolated and identified using standard microbiological techniques. Ascorbic acid, malic acid, lactic acid, and sugar concentrations will be analyzed using high performance liquid chromatography (HPLC). Total antioxidant activity will be determined using a photochemical luminescence method. Collaborators at the University of Hawaii, College of Pharmacy will test samples of noni juice for bioactivity and toxicity in various cell culture assays. Research was conducted to determine the physiology of noni fruit ripening, as well as the chemical and microbial changes that occur during noni fermentation. Ripening noni fruit had a non-climacteric respiratory pattern with an average rate of 34 mg CO2/kg/hr and no detectable ethylene production. Chemical and microbial profiles differed for fresh and fermented juice. Bacterial populations remained low from 0 to 35 days of fermentation. However the fungus, Mucor circinelloides f. circinelloides was consistently isolated from noni juice produced from fruit collected in arid and high-rainfall districts of Hawaii island. M. circinelloides was confirmed as a wound pathogen of noni fruit for the first time. Mucor populations peaked after 14 days of fermentation, coincident with an increase in head space CO2 to 25% and a decline in pH and total soluble solids. These changes preceded the highest juice yield at 21 days. Large volumes of noni juice were partitioned into aqueous, n- butanol, and ethyl acetate fractions to assess antitumor activity in animal studies conducted by collaborators. The results confirmed an earlier report that fermented noni juice has antitumor activity, and also demonstrated that the n-butanol fraction contained the active components. The lead scientist monitors progress through regular meetings with cooperators, and through direct supervision of the research project and participation in research activities.
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Progress 10/01/09 to 09/30/10
Outputs
Progress Report Objectives (from AD-416) 1. Quantify the compositional and microbial changes of noni juice during the fermentation period. 2. Develop a standard method for processing fermented noni juice. Approach (from AD-416) Physiologically mature, ripe noni fruit will be harvested from commercial orchards on the island of Hawaii. Fruit will be washed and placed into sterilized glass jars with sealed lids and allowed to age for 10-14 days at 25�C. A second batch will be aged for up to 30 days. Juice will be pasteurized at 85�C in a water bath, cooled, and frozen. Subsamples of juice will be collected throughout the fermentation process to conduct compositional and microbial analyses. Prominent yeast, bacteria, and fungi will be isolated and identified using standard microbiological techniques. Ascorbic acid, malic acid, lactic acid, and sugar concentrations will be analyzed using high performance liquid chromatography (HPLC). Total antioxidant activity will be determined using a photochemical luminescence method. Collaborators at the University of Hawaii, College of Pharmacy will test samples of noni juice for bioactivity and toxicity in various cell culture assays. Documents Trust with Patient First C Foundation. Log 39444. This report documents research conducted under a Trust Agreement between ARS and the Patient-First Cancer Foundation. Research was conducted towards determining the quality, safety and bioactivity of processed noni juice. Noni fruit were harvested from Kawaihae, HI and fermented in sterile, covered glass jars for up to 4 weeks. Prominent micro-organisms present in fresh and fermented batchs of noni juice were isolated and characterized. Fresh (non-pasteurized) noni juice contained Enterobacteriaceae bacteria (Serratia marcescens, Enterobacter cloacae, and E. sakazakii). Fermented (non-pasteurized) noni juice harbored non- Enterobacteriaceae strains (Aeromonas sp. and Pasteurella sp.) and a fungal endophyte which were not present in fresh juice. No micro- organisms were recovered in pasteurized noni juice samples. Pasteurized fermented noni juice showed significant antitumor activity in mice by collaborators, and tests were begun to identify the active fraction(s) of partitioned noni juice. The ADODR monitors progress through regular meetings with cooperators, and through direct supervision of the research project and participation in research activities.
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