Impact of pH and High-Pressure Pasteurization on the Germination and Development of Clostridium perfringens Spores under Hyperbaric Storage versus Refrigeration.

This study aimed to evaluate hyperbaric storage at room temperature (75-200 MPa, 30 days, 18-23 °C, HS/RT) on Clostridium perfringens spores in brain-heart infusion broth (BHI-broth) at pH 4.50, 6.00, and 7.50 and coconut water (pH 5.40). Both matrices were also pasteurized by high pressure processing (600 MPa, 3 min, 17 °C, HPP) to simulate commercial pasteurization followed by HS, in comparison with refrigeration (5 °C, RF). The results showed that, at AP/RT, spores' development occurred, except at pH 4.50 in BHI-broth, while for RF, no changes occurred along storage. Under HS, at pH 4.50, neither spore development nor inactivation occurred, while at pH 6.00/7.50, inactivation occurred (≈2.0 and 1.0 logs at 200 MPa, respectively). Coconut water at AP/RT faced an increase of 1.6 logs of C. perfringens spores after 15 days, while for RF, no spore development occurred, while the inactivation of spores under HS happened (≈3 logs at 200 MPa). HPP prior to HS seems to promote a subsequent inactivation of C. perfringens spores in BHI-broth at pH 4.50, which is less evident for other pHs. For HPP coconut water, the inactivation levels under HS were lower (≈2.0 logs at 200 MPa). The Weibull model well described the inactivation pattern observed. These results suggest that HS/RT can be simultaneously used as a tool to avoid C. perfringens spores' development, as well as for its inactivation, without the application of high temperatures that are required to inactivate these spores.

Effect of pulsed electric field processing on the quality characteristics and enzyme activity of tender coconut water.

Tender coconut water (TCW) is a natural drink rich in natural electrolytes, minerals, salts and sugars; it has good health benefits. But, its shelf-life is very limited because of the active nature of enzymes present in it when exposed to air. Therefore, the processing of TCW is necessary to inactivate the enzymes. So, this study aims to observe the effect of various process parameters of pulsed electric field (PEF) on the quality parameters of TCW. For the treatment of TCW with PEF, a full-factorial design of experiments was followed with process parameters such as three levels of electric field intensity (8, 12, and 16 kV/cm), two levels of pulse width (PW) (50 and 70 µs), and six levels of the number of pulses (2000 to 12,000 pulses) were considered at a constant pulse OFF time of 75 ms. PEF treatment did not significantly change pH, total soluble solids, and viscosity. However, it significantly affected vitamin C, colour, and total and reducing sugars. PEF treatment significantly enhanced the total phenolic content and antioxidant activity by 23.17% and 42.49%, respectively. At the same time, significant inactivation of polyphenol oxidase (100%) and peroxidase (60.2%) was observed at PEF treatment conditions of 16 kV/cm, 70 µs PW, and 12,000 pulses. Moreover, no significant change in the sensory acceptability of PEF-treated TCW (16 kV/cm, 70 µs PW, 12,000 pulses) when compared to the untreated/fresh TCW, which is a promising sign.

Enhancing bacterial cellulose production of Komagataeibacter nataicola through fermented coconut water by Saccharomyces cerevisiae: A metabonomics approach.

Nata de coco, an edible bacterial cellulose (BC) product, is a traditional dessert fermented in coconut water. Production of Nata de coco by Komagataeibacter nataicola is enhanced by pre-fermented coconut water, but its instability is a challenge. Here, BC production by K. nataicola Y19 was significantly improved by Saccharomyces cerevisiae 84-3 through shaping the metabolite profile of the coconut water. Different fermentation time with S. cerevisiae 84-3 resulted in distinct metabolite profiles and different promoting effect on BC yield. Compared to unfermented coconut water, coconut water fermented by S. cerevisiae 84-3 for 1d and 7d enhanced BC yield by 14.1-fold and 5.63-fold, respectively. Analysis between unfermented coconut water and 1d-fermented coconut water showed 129 significantly different metabolites, including organic acids, amino acids, nucleotides, and their derivatives. Prolonged fermentation for 7d changed levels of 155 metabolites belongs to organic acids, amino acids, nucleotides and their derivatives. Spearman correlation analysis further revealed that 17 metabolites were positively correlated with BC yield and 21 metabolites were negatively correlated with BC yield. These metabolites may affect energy metabolism, cell signaling, membrane integrity, and BC production by K. nataicola Y19. The further verification experiment gave the view that BC yield was not only closely related to the types of metabolites but also the concentration of metabolites. This study provides a novel theoretical framework for a highly efficient BC fermentation system utilizing stable fermented coconut water mediums.

Identification of the Microbiota in Coconut Water, Kefir, Coconut Water Kefir and Coconut Water Kefir-Fermented Sourdough Using Culture-Dependent Techniques and Illumina-MiSeq Sequencing.

The principal objective of this study was to isolate and identify the microorganisms present in commercial kefir grains, a novel kefir-fermented coconut water (CWK) and a novel coconut water kefir-fermented sourdough using phenotypic identification and Sanger sequencing and examine the microbial diversity of CWK and CWK-fermented sourdough throughout the fermentation process using the MiSeq Illumina sequencing method. The phenotypic characterisation based on morphology identified ten isolates of LAB, five AAB and seven yeasts from kefir (K), CWK and CWK-fermented sourdough (CWKS). The results confirm the presence of the LAB species Limosilactobacillus fermentum, Lactobacillus. plantarum, L. fusant, L. reuteri and L. kunkeei; the AAB species Acetobacter aceti, A. lovaniensis and A. pasteurianus; and the yeast species Candida kefyr, Rhodotorula mucilaginosa, Saccharomyces cerevisiae, C. guilliermondii and C. colliculosa. To the best of our knowledge, the identification of Rhodotorula from kefir is being reported for the first time. This study provides important insights into the relative abundances of the microorganisms in CWKS. A decrease in pH and an increase in the titratable acidity for CWK- and CWK-fermented sourdough corresponded to the increase in D- and L-lactic acid production after 96 h of fermentation. Significant reductions in the pHs of CWK and CWKS were observed between 48 and 96 h of fermentation, indicating that the kefir microorganisms were able to sustain highly acidic environments. There was also increased production of L-lactic acid with fermentation, which was almost twice that of D-lactic acid in CWK.

Understanding the maturity of coconut water through 1H NMR profiling and MPAES analyses.

This study investigated the relationship between coconut maturity stages and the sugar, amino acid, and mineral profiles of coconut water (CW). Metabolite profiles were analysed using 1H NMR, covering glucose (G), fructose (F), sucrose (S), reducing sugars (RS), total sugars (TS), amino acids, and organic acids. Mineral composition was measured using Microwave Plasma Atomic Emission Spectroscopy (MPAES). The results revealed distinct metabolite and mineral profiles across different maturity stages. Immature CW had high G/F and RS/TS ratios but low S/G ratios. Conversely, mature CW showed decreased G/F and RS/TS ratios but an increase in S/G. Mineral analysis revealed potassium as the predominant mineral in CW, peaking in the youngest stage and declining with maturity. Sodium, magnesium, and calcium showed a similar pattern, with higher concentrations in early than in later stages. The study identifies the age of 9-10 months as optimal stages for selecting tender coconut water.

Organic Supplementation of Vaccinium corymbosum Micropropagation Media.

The best Vaccinium corymbosum plant growth under in vitro conditions can be achieved by using the right composition and pH of the medium. For the initial phase of in vitro culture, a combination of cytokinins-mostly zeatin-can usually be used. Organic supplementation of the medium enables the use of a replacement for the expensive natural cytokinin used in micropropagation of highbush blueberry. This chapter describes the experiments with silicon Hydroplus™ Actisil (Si), coconut water (CW), and different pH (5.0; 5.5, and 6.0) as a stress factor. The addition of 200 mg dm-3 silicon solution and 15% coconut water strongly stimulated highbush blueberry plant growth in vitro. Moreover, silicon solution benefits the negative effects of higher pH of the medium used for micropropagation of V. corymbosum. Maximum vegetative development of blueberry explants was obtained at pH 5.

Fabrication of freshness indicators based on methylcellulose-containing color indicator solutions for monitoring the quality of coconut water.

This is the first study to apply intelligent packaging to coconut water. The purpose of this study was to determine the best color indicator solution for making freshness indicator labels based on methylcellulose along with the color change profile of coconut water during storage at room temperature. Three color indicator solutions were used, namely phenol red, bromothymol blue, and methyl red, which were then continued with the fabrication of freshness indicator labels based on methylcellulose from each of these color indicator solutions and applied to coconut water at 25 °C room temperature storage for 24 h with observations every 4 h in the form of pH, total dissolved solids, total acid, turbidity, total microbes, CO2 gas, O2 gas, and freshness indicator label color changes. The values of pH, total soluble solids, and O2 gas decreased with storage time, whereas the values of total acid, turbidity, total microbes, and CO2 gas continued to increase. The methylcellulose-based phenol red freshness indicator label provides the best color change profile that matches the freshness condition of coconut water, namely purplish red (fresh), orange (immediately consumed), and yellow (damaged) so that it can be used as intelligent packaging to monitor the quality of coconut water.

Coconut (Cocos nucifera (L.)) Water Improves Glucose Uptake with Concomitant Modulation of Antioxidant and Purinergic Activities in Isolated Rat Psoas Muscles.

The present study investigated the effect of coconut water on glucose uptake and utilization, and metabolic activities linked to hyperglycemia in isolated rat psoas muscles. Coconut water was subjected to in vitro antioxidant and antidiabetic assays, which cover 2,2'-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity, ferric reducing antioxidant power (FRAP), and inhibition of α-glucosidase and α-amylase activities. Psoas muscles were isolated from male Sprague Dawley rats and incubated with coconut water in the presence of glucose. Control consisted of muscles incubated with glucose only, while normal control consisted of muscles not incubated in coconut water and/or glucose. The standard antidiabetic drug was metformin. Incubation with coconut water led to a significant increase in muscle glucose uptake, with concomitant exacerbation of glutathione level, and SOD and catalase activities, while suppressing malondialdehyde level, and ATPase and E-NTDase activities. Coconut water showed significant scavenging activity against DPPH, and significantly inhibited α-glucosidase and α-amylase activities. LC-MS analysis of coconut water revealed the presence of ellagic acid, butin, quercetin, protocatechuic acid, baicalin, and silibinin. Molecular docking analysis revealed potent molecular interactions between the LC-MS-identified compounds, and AKT-2 serine and PI-3 kinase. These results indicate the potential of coconut water to enhance glucose uptake, while concomitantly improving antioxidative and purinergic activities. They also indicate the potential of coconut water to suppress postprandial hyperglycemia. These activities may be attributed to the synergistic effects of the LC-MS-identified compounds.

Chemical Components, Nutritional Value, Volatile Organic Compounds and Biological Activities In Vitro of Coconut (Cocos nucifera L.) Water with Different Maturities.

The differences in chemical components, nutritional value, volatile organic compounds, antioxidant activity and α-glucosidase inhibiting capacity in vitro in coconut waters with different maturities (8, 10, and 12 months after pollination and germination height below 10 cm were named CW-8, CW-10, CW-2, and MCW, respectively) from the tall coconut variety were compared and analyzed. Results showed that as the maturity increased, the ash and reducing sugar in coconut water gradually decreased, while the protein content and fatty acids continued to increase. Potassium, phosphorus, and sodium in four coconut waters showed a trend of first increasing and then decreasing, and CW-12 had the highest content of 2133.85 mg/kg, 239.74 mg/kg, and 310.75 mg/kg, respectively. The volatile organic compounds (VOCs) present in higher amounts are alcohols and esters in coconut waters, among which 2-methylbutyl acetate, ethyl acetate monomer, and 2-methyl-1-propanol dimer were the characteristic volatile substances that distinguish MCW from the other three coconut waters. MCW has the best DPPH-scavenging and ferrous-ion-chelating ability (87.39% and 7.65%), while CW-8 had the highest hydroxyl and ABTS radicals scavenging rate (97.31% and 83.48%) and α-glucosidase inhibitory rate (81.36%). These results can provide support for the differential and high-value utilization of coconut water with different maturities.

Comparative key aroma compounds and sensory correlations of aromatic coconut water varieties: Insights from GC × GC-O-TOF-MS, E-nose, and sensory analysis.

Aroma is a key criterion in evaluating aromatic coconut water. A comparison regarding key aroma compounds and sensory correlations was made between Thailand Aromatic Green Dwarf (THD) and Cocos nucifera L. cv. Wenye No. 4 coconut water using E-nose and GC × GC-O-TOF-MS combined with chemometrics. Twenty-one volatile components of coconut water were identified by GC × GC-O-TOF-MS, and 5 key aroma compounds were analyzed by relative odor activity value and aroma extract dilution analysis. Moreover, the combination of the E-nose with orthogonal partial least squares was highly effective in discriminating between the two coconut water samples and screened the key sensors responsible for this differentiation. Additionally, the correlation between volatile compounds and sensory properties was established using partial least squares. The key aroma compounds of coconut water exhibited positive correlations with the corresponding sensory properties.

Exploring the biotic and abiotic drivers influencing nata de coco production by Komagataeibacter nataicola in pre-fermented coconut water.

In China and Southeast Asia, pre-fermented coconut water is commonly used for the production of nata de coco, a jelly-like fermented food that consists of bacterial cellulose (BC). The inherent natural fermentation process of coconut water introduces uncontrollable variables, which can lead to unstable yields during BC production. This study involved the collection of spontaneously pre-fermented coconut water over a five-month production cycle. The aim was to evaluate the microbiota and metabolite profile, as well as determine its impact on BC synthesis by Komagataeibacter nataicola. Significant variations in the microbial community structure and metabolite profile of pre-fermented coconut water were observed across different production months, these variations had significant effects on BC synthesis by K. nataicola. A total of 52 different bacterial genera and 32 different fungal genera were identified as potential biotic factors that can influence BC production. Additionally, several abiotic factors, including lactate (VIP = 4.92), mannitol (VIP = 4.22), ethanol (VIP = 2.67), and ascorbate (VIP = 1.61), were found to be potential driving forces affecting BC synthesis by K. nataicola. Upon further analysis, the correlation network indicated that 14 biotic factors had a significant contribution to BC production in three strains of K. nataicola. These factors included 8 bacterial genera, such as Limosilactobacillus and Lactiplantibacillus, and 6 fungal genera, such as Meyerozyma and Ogataea. The abiotic factors lactate, mannitol, and ethanol showed a positive correlation with the BC yield. This study provides significant insights into controlling the fermentation processes of pre-fermented coconut water in industrial settings.

Lactiplantibacillus plantarum and Saccharomyces cerevisiae-Fermented Coconut Water Alleviates Dextran Sodium Sulfate-Induced Enteritis in Wenchang Chicken: A Gut Microbiota and Metabolomic Approach.

In order to investigate the potential mechanisms of probiotic-fermented coconut water in treating enteritis, this study conducted a comprehensive analysis of the effects of probiotic intervention on the recovery from Dextran Sodium Sulfate-induced acute enteritis in Wenchang chicks. The analysis encompassed the assessment of growth performance, serum indicators, intestinal tissue structure, and metagenomic and metabolomic profiles of cecal contents in 60 Wenchang chicks subjected to intervention. This approach aimed to elucidate the impact of probiotic intervention on the recovery process from acute enteritis at both the genetic and metabolic levels in the avian model. The results revealed that intervention with Saccharomyces cerevisiae Y301 improved the growth rate of chicks. and intervention with Lactiplantibacillus plantarum MS2c regulated the glycerophospholipid metabolism pathway and reshaped the gut microbiota structure in modeling chicks with acute enteritis, reducing the abundance of potentially pathogenic bacteria from the Alistipes and increasing the abundance of potentially beneficial species from the Christensenellaceae. This intervention resulted in the production of specific gut metabolites, including Gentamicin C and polymyxin B2, recognized for their therapeutic effects on acute enteritis. The combined intervention of S. cerevisiae Y301 and L. plantarum MS2c not only enhanced growth performance but also mitigated intestinal wall damage and increased the abundance of gut metabolites such as gentamicin C and polymyxin B2, thereby mitigating symptoms of enteritis. Furthermore, this combined intervention reduced the levels of serum immune markers, including IL-10, IL-6, TNF-α, IFN-γ, and D-lactic acid, thus mitigating intestinal epithelial cell damage and promoting acute enteritis recovery. This study provides crucial insights into the mechanisms of action of probiotics and probiotic-fermented coconut water in acute enteritis recovery, offering new perspectives for sustainable farming practices for Wenchang chicken.

Green biorefinery of shrimp shell waste for α-chitin and high-value co-products through successive fermentation by co-lactic acid bacteria and proteolytic fungus.

Green biorefinery process was conducted to extract α-chitin and high-value co-products from shrimp shell waste through microbial fermentation using mature coconut water (MCW) as a sole nutrient source. Symbiotic co-lactic acid fermentation (Co-LAF) by Lactobacillus plantarum and Streptococcus thermophilus produced higher levels of lactic acid (LA) and protease activity than their mono-cultures, which led to greater demineralization (DM) and deproteinization (DP) of shrimp shell powder (SSP). After optimizing Co-LAF through Response Surface Methodology and successive fermentation by an acid-active proteolytic fungus Rhizopus oligosporus, the highest DM of 94.0 ± 0.91 % and DP of 86.7 ± 0.1 % were achieved. Based on FT-IR, XRD, and SEM analysis, the bio-extracted chitin had similar structural characteristics to commercial α-chitin but with better quality. These strategies not only contribute to environmentally-friendly and cost-effective extraction of α-chitin (303 ± 18 mg/g-SSP), but also co-produce LA (57.18 ± 0.89 g/L), acid protease (4.33 ± 0.5 U/mL), bio-calcium (277 ± 12 mg-CaSO4/g-SSP), protein hydrolysate (268 ± 5 mg/g-SSP), and pigments (28.78 ± 1.56 µg/g-SSP).

A systematic review on oral rehydration beverages for treating exerciseassociated dehydrationPart II. The effectiveness of alternatives to carbohydrate-electrolyte drinks.

OBJECTIVE: Dehydration associated with exertion is a commonly encountered condition in the first aid setting, particularly at outdoor sporting events. Part I of this back-to-back review demonstrated that commercial sports drinks can be suggested for effective restoration of fluid balance in dehydrated persons. This systematic review was undertaken to compare alternative liquids, such as milk, beer, and coconut water, with water for effective oral rehydration after prolonged exercise. DATA SOURCES: Cochrane Library, PubMed, and Embase were searched for relevant literature in June 2022. STUDY SELECTION: Controlled experimental and observational studies involving adults and children were included when dehydration was induced by physical exercise and oral rehydration fluids were administered and compared with regular water. No additional food intake accompanied the rehydration drinks or water. Articles in all languages were included if an English abstract was available. DATA EXTRACTION: The study design, study population, intervention, outcome measures, and study limitations were extracted from each included article. DATA SYNTHESIS: Out of 3485 records, 11 studies were included comparing skim or low-fat milk, coconut water, and beer (0-5% alcohol) with water. Four studies showed that drinking skim or low-fat milk, without additional food intake, led to a statistically significant improved volume/hydration status when compared with drinking water. In three trials, no significant differences were shown at multiple timepoints for outcomes related to volume and hydration status following rehydration with fresh coconut water compared with water. Lastly, there is insufficient evidence to recommend beer for rehydration (0-5% alcohol). CONCLUSIONS: Consuming skim or low-fat cow's milk without additional food as compared with water appears to improve volume/hydration status in persons with exercise-induced dehydration. However, evidence is of very low certainty and should be interpreted with caution.

Effect of feed enriched by products formulated from coconut water, palm sap sugar, and mushroom on the chemical composition of feed and carcass, growth performance, body indices, and gut micromorphology of giant gourami, Osphronemus goramy (Lacepède, 1801), juveniles.

Background: Giant gourami, Osphronemus goramy is the most important freshwater fish species produced by aquaculture in Indonesia. This study aimed to the effects of various newly formulated products on the amino acid composition of the diet and whole-body carcass, and to analyse the growth coefficient, body indices, and gut micromorphology. Methods: One hundred gram of palm sap sugar was cooked in 1.1 litre of fresh water for fifteen minutes, to create 1 litre of 11% palm sap sugar solution (after some of it had been boiled off). Two litres of coconut water were then mixed with the litre of palm sugar solution. One litre of this product was added in turn to 2 g of Aspergillus niger (CP2), 2 g of Rhizopus oligosporus (CP3), and 2 g of Saccharomyces cerevisiae (CP4), while freshwater was used as a control (labeled CP1). Aquafeed was added to CP1, CP2, CP3, and CP4, to make diets labeled KP1, KP2, KP3, and KP4. The dosage was 150 ml/kg of feed. Juvenile giant gourami (initial weight 50±0.25 g and length 13.2±0.07 cm) were reared in triplicate net frames (2×1×1 m; water volume 1.5 m 3) in a freshwater concrete pond with a stocking density of 30 juveniles/net. Results: The results supported our hypothesis that different product formulations have a significant effect (P < 0.05) on aquafeed nutrition and the whole-body carcass, growth coefficient, feed utilization, body indices, and gut micromorphology of giant gourami juveniles. The thermal growth coefficient strongly correlated with the daily growth coefficient (r 2 = 92%), condition factor (r 2 = 77%), protein efficiency ratio (r 2 = 75%), while a moderate relationship with the feed intake ( r 2 = 69%). Conclusions: Diet KP3 contains higher total amino acids in diets and carcasses and leads to feed efficiency and better growth for giant gourami.

Green SPIONs as a novel highly selective treatment for leishmaniasis: an in vitro study against Leishmania amazonensis intracellular amastigotes.

The main goal of this work was to evaluate the therapeutic potential of green superparamagnetic iron oxide nanoparticles (SPIONs) produced with coconut water for treating cutaneous leishmaniasis caused by Leishmania amazonensis. Optical and electron microscopy techniques were used to evaluate the effects on cell proliferation, infectivity percentage, and ultrastructure. SPIONs were internalized by both parasite stages, randomly distributed in the cytosol and located mainly in membrane-bound compartments. The selectivity index for intracellular amastigotes was more than 240 times higher compared to current drugs used to treat the disease. The synthesized SPIONs showed promising activity against Leishmania and can be considered a strong candidate for a new therapeutic approach for treating leishmaniases.

Evaluation of Viability of Periodontal Ligament Cells using Propolis, Coconut Water, Aleo Vera, and Soy Milk: An In vitro Comparative Study.

Objectives: This research was done to assess the periodontal ligament cells viability using propolis, coconut water, aloe vera, and soy milk storage media. Materials and Methods: Periodontal ligament cells were taken from freshly extracted teeth and stored into four groups of storage media: Group A-propolis, Group B-coconut water, Group C-aloe vera, and Group D-soy milk. Later, the cell viability was assessed at 1, 12, and 24 hours. Results: The periodontal ligament cell viability was significantly greater in propolis, followed by coconut water, aloe vera, and soy milk. Conclusion: We found that propolis, coconut water, aloe vera, and soy milk can be used as an effective storage media. These are naturally occurring and easily available storage medium.

Single-Blind Randomized Controlled Trial: Comparative Efficacy of Dark Chocolate, Coconut Water, and Ibuprofen in Managing Primary Dysmenorrhea.

Dysmenorrhea, the pain experienced by women during menstruation, affects a significant proportion of women worldwide and often leads to decreased productivity. Various pharmacological and non-pharmacological treatments are available for pain relief, but information on their effectiveness, particularly regarding green coconut water, dark chocolate, and Ibuprofen, remains limited. This study aimed to compare the effectiveness of green coconut water, dark chocolate bars, and Ibuprofen in reducing the intensity of primary dysmenorrhea. In this research, a randomized controlled trial with a quantitative design was conducted, involving 45 participants randomly assigned to receive 330 mL of green coconut water, 35 g of 70% dark chocolate, or 400 mg Ibuprofen. The interventions were administered on the first day of menstruation when dysmenorrhea symptoms typically occur in subjects. This study used a single-dose approach to evaluate the immediate impact of each treatment. The subjects were instructed to consume the given interventional product within 15 min. The pain intensity was measured using a Numeric Rating Scale before the intervention and 2 h after the subjects finished consuming the interventional product. The multivariate Kruskal-Wallis test revealed a significant difference in effectiveness among the three interventions (p < 0.05). The study found that Ibuprofen was the most effective intervention compared to the other interventions. These findings contribute to understanding the treatment options for primary dysmenorrhea and emphasize the efficacy of Ibuprofen (trial registration: ClinicalTrials.gov: NCT05971186).

Discrimination and characterization of different coconut water (CW) by their phenolic composition and volatile organic compounds (VOCs) using LC-MS/MS, HS-SPME-GC-MS, and HS-GC-IMS.

Three varieties of coconut (Cocos nucifera L.) water (CW) at two maturity stages were investigated for physicochemical and nutritional properties. The profile of phenolic compounds and volatile organic compounds (VOCs) was determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS), headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS), and headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS). Most of the properties of CW changed significantly with maturity rather than variety. The five most relevant phenolic compounds in CW were chlorogenic acid, 4-hydroxy-3,5-dimethoxycinnamic acid, L-epicatechin, and procyanidins B2 and B1. Variety played a more important role in phenolic composition than maturity, and Wenye No. 4 can be distinguished from other two varieties. Alcohols and esters were the main VOCs in CW identified by HS-GC-IMS and HS-SPME-GC-MS, respectively. Five and four compounds (VIP scores > 1) were characteristic compounds for CW by HS-GC-IMS and HS-SPME-GC-MS, respectively. The VOCs of Wenye Nos. 2 and 3 were more similar than those of Wenye No. 4. These findings could provide useful information for the selection of raw materials of CW used for different industrial purposes.

Canine sperm vitrification with nonpermeable cryoprotectants and coconut water extender.

This study was aimed to assess the efficiency of coconut water extender with addition of soy lecithin and sucrose as nonpermeable cryoprotectants for canine semen vitrification, using a simple method that yields a high survival rate of spermatozoa for clinical use. Twelve ejaculates from 12 adult normozoospermic dogs were collected separately by digital manipulation and only the second semen fraction was used in this study. After evaluation of volume, concentration, viability, total and progressive motility, velocity parameters and morphology, semen was diluted with a coconut water extender (50% (v/v(volume per volume)) coconut water, 25% (v/v) distilled water and 25% (v/v) 5% anhydrous monosodium citrate solution) with addition of soy lecithin and fructose at 1% and 0.25M sucrose until final concentration of 100x106 spermatozoa/ml. After equilibration at 5ºC for 60 minutes, semen was vitrified by "direct dropping method" into liquid nitrogen in spheres with a volume of 30 µl. After a week of storage the spheres were devitrified as three of them were dropped into 0.5 mL of CaniPlus AI medium (Minitüb, Germany), which was previously warmed in a water bath at 42ºC for 2 minutes and evaluated about the above mentioned parameters. It was found that vitrification resulted in a lower percentage of viable sperms, normal morphology, total and progressive motilities (p<0.05), but most of velocity parameters (VCL, VSL, VAP, LIN, ALH and BCF) did not differ (p>0.05) compared to fresh semen samples. In conclusion, our results demonstrate that vitrification with coconut water extender with addition of 1% soy lecithin and 0.25M sucrose as cryoprotectants, has an excellent potential for routine canine sperm cryopreservation.