Breaking the spores of Ganoderma lucidum by fermentation with Lactobacillus plantarum

Ganoderma lucidum has been utilized in medical field from ancient time. In recent studies, it is found that spores of G. lucidum consists of several bioactive substances which are much abundant than in the fruiting body. A hard sporoderm limits the absorption of these active compounds. Therefore, it is necessary to break down the sporoderm to allow complete absorption. The spore breaking methods which have been developed are physical smashing, ultrasonic, high pressure and enzymes application. A few  limitations have come forward; they are low yield, high cost, high technology tools requirement and loss valuable substances during production. In this paper, fermentation of G. lucidum with Lactobacillus plantarum was applied to break down the sporoderm. Scanning electron microscope (SEM) was used to characterize the spores. The broken spores were found on the 3rd day and complete breaking on the 5th day of fermentation. Lactic acid, acetic acid and combination of these acids did not cause the spore to break down. This technique is simple, less cost. It can be used to produce fermented juices with the benefit from G. lucidum spores and lactic acid bacteria. Mechanism of spore break downby lactic acid bacteria is further investigated.Key words: Ganoderma lucidum, break spore, Lactobacillus plantarum, lactic acid bacteria, fermentation

Factors affecting methanol content of fermented plant beverage containing Morinda citrifolia

Pectin extraction of fresh Morinda citrifolia Linn or Noni using HCl at various concentrations was studied. The highest pectin yield (9.89% w/w) was achieved by using 20 mM HCl. Pectinmethylesterase (PME) activity and specific activity of raw M. citrifolia were 0.276 (μmol/ml. min) and 1.48 (Units/mg), respectively whereas, in the ripen M. citrifolia, they were 1.006 (μmol/ml. min) and 2.96 (Units/mg), respectively. Four formulas (F1, F2, F3 and F4) of the fermented plant beverage (FPBs) were prepared by varying material size (blended and diced) and sterilization process [72°C and using potassium metabisulfite (KMS)] using factorial design. Soluble pectin in FPBs using blended material (F3 and F4) was higher than in FPBs using diced material (F1 and F2). Soluble pectin in FPBs using pasteurization (F1 and F3) was also higher than FPBs using KMS (F2 and F4). At 24 h fermented period, methanol concentration in FPBs using blended material (F3 and F4) was higher than FPBs using diced material (F1 and F2). Methanol concentration in FPBs using KMS (F2 and F4) was also higher than FPBs using pasteurization (F1 and F3). After 24 h fermentation time, methanol concentration in FPBs using blended material (F3 and F4) could not be detected until the end of the fermentation whereas, in FPBs using diced material, methanol was found throughout the fermentation (F1 and F2). The study indicates that the most influencial factor on methanol production in FPBs was raw material size. Other factors were sterilization, PME and pectin, respectively. Thus, the recommended procedure of the fermentation of plant beverage was the use of blended raw M. citrifolia and pasteurization at 72°C for 15 s. These conditions prevent methanol production in FPBs. Therefore, quality and safety of the FPBs will be accepted by consumers.Keywords: Morinda citrifolia, methanol, pectin, pectinmethylesterase (PME), fermented plant beverage (FPB)African Journal of Biotechnology Vol. 12(27), pp. 4356-436

LACTOBACILLUS-FERMENTED PLANT JUICE AS A POTENTIAL INGREDIENT IN COSMETICS: FORMULATION AND ASSESSMENT OF NATURAL MOUTHWASH

ABSTRACTObjective: Oral care cosmetics are essential for all populations and are systematically used to treat oral problems. The chemicals free natural cosmeticsare the choice of many people. Thus, this study was aimed to formulate and to assess the natural mouthwash (MW) solution from LactobacillusfermentedThai medicinal plants juice.Methods: The selected (betel, green tea, clove, black galingale, mangosteen, and noni) plant juices were subjected to Lactobacillus plantarum mediatedfermentation. The fermented plant juices (FPJ) were formulated into MW solution with different concentrations of peppermint oil. MW formulationswere assessed for physical appearance, stability, and anti-microbial activities.Results: About 2% of peppermint oil in FPJ was found as organoleptically optimum. The pH and refractive indexes of the MWs were not affectedduring storage and stability assessments. All the FPJ-MWs formulations showed antimicrobial activity against Group A Staphylococcus, and other oralpathogens - Escherichia coli, Streptococcus aureus, and Pseudomonas aeruginosa. Moreover, black galingale, mangosteen, and noni based MW formulasalso exhibited anti-candida activity. The MW made from fermented black galingale (Kaempferia parviflora) juice was the most potent antimicrobialformulation with excellent physical stability.Conclusion: The study concluded that fermented plant-herbal juices can be used as natural MW recipe with 2% of peppermint oil to improve theflavor and aroma. The formulations were stable, free of microbial contamination, and also exhibited antimicrobial activity. Further extended stabilitystudy and clinical trials are necessary to develop a commercial FPJ-based natural MW recipe.Keywords: Fermentation, Formulation, Lactobacillus, Mouthwash

PHENETHYL ALCOHOL IS AN EFFECTIVE NON-TRADITIONAL PRESERVATIVE AGENT FOR COSMETIC PREPARATIONS

Objective: Preservatives are used in the cosmetic products to protect the potential growth of microbes, therefore to prolong the shelf-life of products and to protect the consumer from infections. However several preservatives can cause various health problems, and the safety profiles of those preservatives are still unclear. Many natural substances are used in the cosmetic products to substitute the traditional preservatives. The present study deals with the evaluation of conservative nature of phenethyl alcohol (PEA) in three cosmetic formulations (emulsion, cleansing, and conditioner).Methods: Three different concentration of PEA (0.3, 1, and 2.5 %) were used in cosmetic formulations. The physical appearance of the formulas was assessed manually, and the antimicrobial nature of PEA and PEA-containing cosmetic formulations were evaluated by agar well plate assay. Results: The use of PEA has not affected the physical appearance, and quality of the formulations, except the high concentration of PEA in the cleansing solution, which reduced the foam formation. The minimal required concentration of PEA in emulsions and cleansings were 1.0% and 2.5% in the conditioners. All cosmetic preparations were subjected to antibacterial and antifungal evaluation. The alkaline pH (> 8) affected the antimicrobial activity of PEA in a cosmetic product.Conclusion: The results suggested that PEA is a potent non-traditional preservative for the cosmetic formulations especially in the emulsion, cleansing solutions, and conditioners

EVALUATION OF PHARMACOLOGICAL STABILITY OF PERILLA OIL AND PERILLA OIL CAPSULE

Objective: The present study explained the effect of storage conditions on the stability (total phenolic content (TPC), acid and peroxide values, and antioxidant capacity) of free perilla oil (PO) and capsulated PO (POC). Methods: PO from Mae Hong Son cultivar was used for making POC. The capsules were prepared by Nature Nutri Co., Ltd., Pathum Thani, Thailand. PO and POC samples were stored in different containers (plastic, glass, clear, and amber bottles) at various temperatures (4, 30, and 40°C) for 3 months. The physical changes (color, odor, sedimentation, and separations), TPC, and antioxidant capacity of the samples were assessed by organoleptic, Folin–Ciocalteu colorimetric, and 1,1-diphenyl-2-picrylhydrazyl and 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid assay methods, respectively. The acid and peroxide values of the samples were also studied by the titration method. Results: The color and odor of PO were not changed during storage, and also, there was no sedimentation and layer separation. The acid values were not significantly changed during the storage period, but the peroxide values were significantly increased in PO samples stored at 40°C while no changes were observed in POC. The TPC and antioxidant capacity of the samples were not influenced by any of the storage conditions. Conclusion: The results suggested that POC was relatively more stable than free oil in terms of peroxidation

INFLUENCE OF EXTRACTION PROCESS ON YIELD, TOTAL PHENOLIC CONTENT, AND ANTIOXIDANT PROPERTIES OF AVOCADO (PERSEA AMERICANA MILL.) OIL AND STABILITY ASSESSMENT

Objective: The present study evaluated the impact of drying and storage conditions on the quality of the avocado oil. Methods: The fresh avocado was obtained from local farmers of Chiang Mai province, Thailand. The avocado pulp was collected and blended. The blended samples were dried at different temperatures (60, 80, and 100°C) and the oil was extracted by manual or mechanical pressing. The avocado oil samples were stored in clear and amber glass bottles at various temperatures (4, 30, and 40°C) for 3 months. The organoleptic, Folin–Ciocalteu colorimetric, 1,1-diphenyl-2-picrylhydrazyl and 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid assay, and titration methods were used to measure the physical changes (color, odor, and precipitation), total phenolic content (TPC), antioxidant activity, and acid value (AV) and peroxide values (PVs) of the samples, respectively. Results: The manual squeezing and mechanical pressing methods yielded about 1.17 and 24.91% of avocado oil from fresh, undried avocado flesh, and the pH of both samples was 5. The AV of manually squeezed and mechanically pressed oil samples was 2.91 and 2.88 mg KOH/g of oil, respectively. The PV of manually squeezed and mechanically pressed oil samples was 7.96 and 8.03 milliequivalent of oxygen per Kg of oil, respectively. The quality of the avocado oil dried at 60°C was high compared to other samples regarding TPC and antioxidant capacity. The storage of oil samples for 3 months at various temperatures and in different containers has not affected the quality significantly, except at 40°C. Conclusion: The study proposed that the drying process at more than 80°C interferes with the quality of the avocado oil, and the storage conditions have a greater impact on the stability of the avocado oil. The storage at a low temperature in the absence of light may extend the shelf life of the avocado oil

IMPACT OF PHYSICAL TREATMENTS ON STABILITY AND RADICAL SCAVENGING CAPACITY OF ANTHOCYANIDINS

Objective: The aim of the study was to evaluate the chemical stability and antioxidant ability of selected anthocyanidin (ACN) upon various commonly used physical treatments.Methods: Pure compounds of representative ACNs (cyanidin, peonidin) were subjected to microwave, heat, and sonication treatments followed by analyzing the rate of degradation by LC-MS. The changes in the antioxidant ability of ACNs were also assessed by DPPH and ABTS assay.Results: All the tested treatment strategies accelerated the degradation and diminished the antioxidant capacity of pure ACN, more specifically heat exposure cause ~ 90% of degradation and ~3 fold reduction in antioxidant capacity. About 91.34% and 87.73% of cyanidin and peonidin degradation were documented after heat treatment, respectively. Relatively sonication has not accelerated the ACN debasement, but significant level of degradation (p<0.05) was observed.Conclusion: The study results suggested that the maximum concern is required for the selection of the method of the degerming process during the production of precious formulations. This study revealed that microwave and sonication processes are better than dry heat based aseptic methods for pure ACNs based product, especially in pharmaceuticals with respect to the stability and bioactivity of ACN.Â

ASSESSMENT OF BIOLOGICAL SAFETY OF FERMENTED PHYLLANTHUS EMBLICA FRUIT JUICE

Objective: The present study evaluated the subchronic toxicity of Lactobacillus mediated fermented Phyllanthus emblica fruit juice (FPJ) using a rat as a model system.Methods: FPJ was prepared, and estimated the changes in pH by pH meter, and microbial load by a plating method. Rats were fed with different dose of FPJ for 60 days. The changes in the body mass were noted. The blood and organs of the experimental rats were collected, after 60 days of intervention. Then, they were analyzed for the selected hematological and biochemical parameters by following standard hospital protocols.Results: The pH of FPJ after 30 days of fermentation was 3.16. FPJ was rich in probiotic Lactobacillus spp. (7.23 Log CFU per mL) without contamination. The supplementation of FPJ was not significantly affected the body weight of the experimental animals, except the female rats in posteffective dose (PED) group showed significant changes (20.83±8.49 g) compared to control (40±17.22 g). The internal organs of the rat were not affected by the FPJ supplementation. The changes observed in blood urea nitrogen, creatinine, cholesterol, triglyceride, aspartate aminotransferase; alanine aminotransferase, and alkaline phosphatase level of experimental rats, both male and female, were not significantly differed from the respective controls. The average of lymphocytes level was significantly increased in continuous dosing group of males and females. Interestingly, the increase in red blood cell and hemoglobin (HGB) were statistically significant for ED group and PED in both sexes, except for females with no effect on HGB content.Conclusion: The prepared FPJ was enriched with probiotic Lactobacillus spp. The supplementation of FPJ (up to 9 mL/kg/day) for 60 days was not significantly influenced the body weight, internal organs, biochemical and hematological parameters of experimental rats (both male and female). The results revealed that FHJ is suitable for the human consumption

ANTHOCYANIN PROFILE AND ITS ANTIOXIDANT ACTIVITY OF WIDELY USED FRUITS, VEGETABLES, AND FLOWERS IN THAILAND

Objective: Anthocyanins are the water-soluble pigments most commonly present in flowers, fruits, and vegetables. Anthocyanins have been reported for is antioxidant, anti-inflammatory, antibacterial properties and other health benefits in human. This study was performed to extract and determine the anthocyanin and to access the antioxidant properties of the most commonly used fruits, vegetables and flowers in Thailand.Methods: Extraction of anthocyanin and its aglycones from the selected samples and determination of anthocyanin and anthocyanidins using reversed-phase HPLC analysis. Determination of total antioxidant ability using ABTS, DPPH and FRAP assay.Results: Maximum distribution of tested anthocyanin and anthocyanidin contents was observed in the extracts of Jambolan plum, Ceylon Spinach, Purple Sweet Potato, Purple Lettuce followed by common plum and Red dragon fruit. The extracts of Ma-Kiang, Purple Eggplant, Red Grape, and Purple Lettuce showed the maximum amount of cyanidin 3-glucoside, delphinidin 3-glucoside, peonidin 3-glucoside, and malvidin 3-glucoside, respectively. Likewise, the maximum concentration of cyanidin, delphinidin, peonidin, and malvidin were found in Common plum, Red Khae, Red Cabbage, and Purple Lettuce, respectively. Ma-Kiang extract exhibited the maximum antioxidant activity followed by Red Cabbage, Common Plum, and Purple Eggplant extract compared to the other extracts.Conclusion: The present study, primarily profiled the anthocyanin content of selected fruits, vegetables, and flowers. Among the tested samples, Ma-Kiang extract showed the high cyanidin 3-glucoside content and antioxidant activity. Further, detailed study on the content of anthocyanins at different climate and geographical conditions and other factors are necessary to develop nutraceutical or cosmetic product with a functional ingredient.Keywords: Anthocyanin, Anthocyanidin, Antioxidant activity

ASSESSMENT OF SUBCHRONIC TOXICITY OF FERMENTED HOUTTUYNIA CORDATA THUNB. USING RODENT MODEL SYSTEM

Objective: The present study was intended to evaluate the subchronic toxicity of Lactobacillus-mediated fermented Houttuynia cordata juice (FHJ) using rodent model system.Methods: FHJ was prepared and the microbial load, lactic acid content, and pH was estimated. Rats were fed with different doses of FHJ for 60 days. The body mass changes were measured during FHJ supplementation. After the treatment period, blood and organs of the experimental rats were collected. The samples were subjected to hematological and biochemical analysis by following standard hospital protocols.Results: The pH of FHJ after 30 days of fermentation was 3.63. The lactic acid content of FHJ was gradually increased and reached 19.70 mg per mL after 30 days of the fermentation process. Lactobacillus load was high in FHJ after 30 days and no Bacillus spp. and yeast were detected in FHJ at any point of fermentation. There were no significant changes in body weight of male and female experimental rats supplemented with FHJ, irrespective of dose. There were no significant treatment-related pathological changes found in any organ of the experimental rats at all tested dose levels when compared with organs in control animals. There were no significant changes observed in red blood cells (RBCs), white blood cells (WBCs), hemoglobin, hematocrits, lymphocyte, and platelets level of male rats of all groups. Whereas, significant (p<0.01) changes were observed in the RBC (1.02±0.26 106/mm3) of female rats in effective dose (ED) group compared to control. Similarly, significant (p<0.01) changes were detected in the WBC level of female rats in high dose (−7.53±0.03 103/mm3), and post-ED group (−8.86±0.75 103/mm3) compared to control. There were no alterations in tested biochemical parameters of experimental rats.Conclusion: The FHJ was rich in probiotic Lactobacillus. The supplementation of FHJ (9 mL/kg/day) for 60 days did not significantly affect the body mass, internal organs, hematological, and biochemical parameters of rats. The results suggested that FHJ is qualified for the human consumption