Hydrolysable Tannins Research Articles

«Алкогольная» ферментация табака в дубовых бочках. Часть 1. Массообмен танина и его косвенное количественное определение методом ВЭЖХ

The quantitative transition of oak tannin into raw tobacco during its fermentation in oak barrels in the gas-liquid medium of cognac distillate was studied. The object of research was Cuban cigar tobacco of the HP variety. The fermentation process was carried out for six months with periodic (1–2 weeks) treatment of the inner surface of the barrel with cognac distillate with an alcohol content of 70 vol.% in an amount of 1,5 cm3/dm2. To obtain data on tannin mass transfer, we developed a method for the quantitative indirect determination of tannin in oak and tobacco raw materials using reverse-phase high-performance liquid chromatography (RP HPLC) through preliminary hydrolysis of tannin to gallic acid. The determination of gallic acid/tannin was carried out on a liquid chromatograph with a spectrophotometric detector (UV) at a wavelength of 280 nm using the external standard method. The analysis of unfermented tobacco raw materials did not reveal the presence of tannin. It has been shown that under the specified conditions, during 6 months of fermentation, diffusion of tannin from oak staves into tobacco is observed in an amount of 0,11% by weight of tobacco. The mass fraction of tannin in oak staves was 0,81%. At the same time, the most intense saturation occurs in the 1st, 4th, and 5th months of fermentation.

Rapid deterioration in buried leather: archaeological implications.

Understanding archaeological leather degradation helps inform economies, crafts, and technologies of historic communities. However, archaeological leather is at high risk of degradation due to deterioration and changes within the burial conditions. This research applied non-destructive FTIR-ATR to experimentally buried vegetable-tanned leather and archaeological leather excavated at the Roman site of Vindolanda, UK to explore survival, destruction, and preservation processes of tanned leather. Analyses focused on observing and monitoring changes in chemical functional groups related to leather tannins, collagen and lipid components following burial. FTIR-ATR results highlighted rapid changes following experimental burial in wet soil, tentatively associated with early onset microbial activity, which targeted readily available lipids but not tightly bound collagen. Prior to burial, differences in structural composition were present in leather spectra based on manufacture; however, following burial in wet soil, FTIR-ATR spectra indicated de-tanning occurs rapidly, especially in waterlogged conditions, with archaeological leather becoming more uniform and similar to untanned leather. Therefore, the comparison of FTIR-ATR results from archaeological leather to experimentally buried leather samples was informative for showing the destructive de-tanning in waterlogged environments. The comparison of FTIR-ATR data from modern unburied leather cannot be compared against archaeological samples. Importantly, despite de-tanning occurring soon after burial, the vegetable-tanning method promoted long-term preservation of leather in wet soil. The observed changes could not be directly associated with the proportion of condensed to hydrolysable tannin, suggesting alternate variables impacted the preservation. Furthermore, mineral components introduced into the leather through the animal skin, tannin material and/or tannin liquid are suggested to contribute to these changes. Crucially a high degree of heterogeneity in error results within the experimentally buried sample material underlined that any changes in collagen ratios cannot be overinterpreted and must be considered within the context of larger datasets.

Exploring the bioactive compounds from berry biowaste: Eugenia involucrata DC. as a study case

Aiming at valorising the Eugenia involucrata residue, the present work aimed in investigating the chemical profile and in vitro bioactivities of the residue (EIR) and its individual parts, i.e., seeds (EIS) and peels (EIP). Chromatographic analyses showed that EIP had the highest content of organic acids, tocopherols, and anthocyanins, whereas EIS was the richest in hydrolysable tannins. bis-HHDP-glucoside isomers and cyanidin-3-O-galactoside were the main detected phenolic compounds. By the cellular antioxidant activity (CAA) assay, the samples showed similar. antioxidant potential with 71%, 77%, and 80% inhibition for EIP, EIR, and EIS, respectively, while EIS demonstrated the best antioxidant activity by thiobarbituric acid reactive substances (TBARS, IC50 = 4.96 μg/mL) and oxidative inhibition assay (OxHLIA, IC50 = 40 μg/mL). The samples were able to inhibit the growth of seven bacteria and two fungi. Between the microorganisms tested, all samples showed a strong antibacterial activity (MIC = 0.156 mg/mL) against Yersinia enterocolitica and Pseudomonas aeruginosa (MIC = 0.156–0.625 mg/mL). Moreover, they also display a high antifungal activity against Arpergillus brasiliensis (MIC = 0.625 mg/mL). Furthermore, only EIR and EIS had anti-inflammatory effect via NO production inhibition (IC50 = 168–259 μg/mL, respectively) and anti-proliferative activity towards four tumour and one non-tumour cell lines. These findings suggest that the biowaste generated during the processing of E. involucrata fruit has potential as a source of bioactive compounds.

Study on the mechanism of natural polysaccharides on the deastringent effect of Triphala extract

This present study investigated the masking effect of high methoxyl pectin, xanthan gum, and gum Arabic on the astringency of the traditional herbal formula Triphala and further examined the mechanism of polysaccharide reducing astringency. Results of sensory evaluation and electronic tongue illustrated that 0.6 % pectin, 0.3 % xanthan gum, and 2 % gum Arabic had a substantial deastringent effect. The polyphenols in Triphala are basically hydrolysable tannins, which with high degree of gallic acylation may be the main astringent component of Triphala. Moreover, the three polysaccharides can combine with β-casein through CO and NH groups to form soluble binary complexes and decrease the secondary structure of β-casein. When polysaccharides were added to the Triphala-protein system, polyphenol-protein precipitation was also diminished, and they were capable of forming soluble ternary complexes. Consequently, the competition between polysaccharides and polyphenols for binding salivary proteins and the formation of ternary complexes help decrease the astringency of Triphala.

Possible Beneficial Effects of Hydrolyzable Tannins Deriving from Castanea sativa L. in Internal Medicine.

Hydrolyzable tannins (HTs) deriving from chestnuts have demonstrated, through numerous studies, the ability to exert multiple beneficial effects, including antioxidant and antimicrobial effects, on the lipid metabolism and cancer cells. The latter effect is very fascinating, since different polyphenols deriving from chestnuts were able to synergistically induce the inhibition of cancerous cells through multiple pathways. Moreover, the main mechanisms by which tannins induce antioxidant functions include: the reduction in oxidative stress, the ability to scavenge free radicals, and the modulation of specific enzymes, such as superoxide dismutase. HTs have also been shown to exert significant antimicrobial activity by suppressing microbial growth. The actions on the lipid metabolism are several, among which is the inhibition of lipid accumulation. Thus, tannins seem to induce a cardioprotective effect. In fact, through various mechanisms, such as the relaxation of the vascular smooth muscle, HTs were proven to be efficient against arterial hypertension. Therefore, the great number of studies in this field prove the growing interest on the utilization of natural bioactive compounds, such as HTs deriving from natural sources or obtained by circular economy models, as potential nutraceuticals or adjuvants therapies.

Pharmacological Property and Cytotoxic Effect Showing Antiproliferative Potency in Human Melanoma Cell Lines (A375) of Combretum racemosum P. Beauv. Leaf and Root Extracts Used in Benin.

Combretum racemosum, a plant from the Combretaceae family, is traditionally used in Benin for various health problems. However, scientific research on Beninese samples of this plant is limited. The aim of this study was to identify and assess the bioactive compounds in the plant's leaves and roots. Initial screening involved analyzing powders derived from these parts for total polyphenols, flavonoids, and both condensed and hydrolyzable tannins. The polyphenolic compounds were analyzed using HPLC-DAD-ESI-MS. To evaluate the plant's antimicrobial properties, the agar diffusion method was employed, while FRAP and DPPH assays were used to determine its antioxidant capacity. For anti-inflammatory activity, the study utilized tests for in vitro protein denaturation inhibition and in vivo acute edema induced by carrageenan. Additionally, an antiproliferative assay was conducted using the human melanoma cell line A375. The analysis revealed the presence of significant polyphenolic compounds in both the leaf and root extracts of C. racemosum. Notably, compounds like Pedunculagin, Vescalagin, Casuarictin, and Digalloyl-glucoside were abundant in the leaves, with Vescalagin being especially predominant in the roots. The study also found that the dichloromethane extracts from the leaves and roots exhibited bactericidal effects on a substantial percentage of meat-isolated strains. Moreover, the antioxidant activities of these extracts were confirmed through FRAP and DPPH methods. Interestingly, the dichloromethane root extract showed strong activity in inhibiting thermal albumin denaturation, while the water-ethanol leaf extract demonstrated significant edema inhibition. Finally, the study observed that C. racemosum extracts reduced cell viability in a dose-dependent manner, with leaf extracts showing more pronounced antiproliferative effects than root extracts. These findings highlight the potential of C. racemosum leaves and roots as sources of compounds with diverse and significant biological activities. In conclusion, C. racemosum's leaves and roots exhibit promising biological activities, highlighting their potential medicinal value.

Exploring In Vitro the Combination of Cistus × incanus L. and Castanea sativa Mill. Extracts as Food Supplement Ingredients against H. pylori Infection.

Cistus spp. have been traditionally used for inflammatory and infectious disorders, including gastrointestinal ailments, in the Mediterranean area. Among them, Cistus × incanus L. is one of the most frequently cited species in the literature for a variety of biological activities which include inflammatory diseases. Cistus spp. aerial parts are rich in polyphenols such as condensed and hydrolysable tannins, procyanidins, and flavonoids, which show gastroprotective activities. The purpose of the present study is to investigate the biological activities of a hydroalcoholic extract from Cistus × incanus L. aerial parts in gastric epithelial cells (GES-1) infected with H. pylori. The extracts inhibited IL-8 and NF-κB induced by H. pylori and showed antibacterial activity after simulated digestion. Since our previous paper reported interesting results on the ability of Castanea sativa Mill. leaf extract to decrease inflammatory conditions in H. pylori-infected gastric cells, the combination of Castanea sativa and Cistus × incanus extracts was also investigated, showing strong anti-inflammatory activity and inhibition of bacterial adhesion. This association of botanicals is proposed herein as a novel food supplement capable of counteracting gastric inflammatory conditions.

The Influence of Tannins, Saponins, and Direct Fed Microbials on In Vitro Rumen Fermentation Characteristics and Microbiota

In-vitro experiments were conducted to determine the impact of tannins, saponins, and DirectFed-Microbials (DFM) on rumen fermentation characteristics. Rumen fluid was collected 2h post feeding from three cannulated crossbred (Angus x Hereford) feedlot steers (450±12.2 kg), adjusted to finishing diet. Rumen fluid from all steers were filtered, combined, and mixed with buffer. Vaccine bottles containing ground finishing diet and appropriate treatments were incubated with rumen fluid mixture (39 ˚C). Treatments consisted of 1) Control; 2) Commercial Tannin (TCH); 3) Hydrolysable Tannin (HT); 4) saponin; 5) TCH+DFM; 6) HT+DFM; and 7) Saponin+DFM. At 12 h post incubation, HT+DFM and Saponin+DFM had lesser (P<0.05) molar proportions of acetic acid when compared to all other treatments. Saponin+DFM had greater (P<0.05) molar proportions of propionic acid and a lesser (P<0.05) acetate: propionate ratio when compared to treatments not containing DFM. Dry matter digestibility and pH were greater (P<0.05) for saponin, TCH+DFM, HT+DFM and saponin+DFM, whereas pH was greater in treatments containing DFM compared to the controls. Percent CO2 was lesser (P<0.05) in saponin and THC+DFM in vitro vessels compared to controls. At 18 h post fermentation total VFA concentrations, molar proportions of butyric and valeric acid, and gas cap N2 percentage were greater (P<0.05) in in vitro vessels containing HT+DFM compared to controls. The percentage of CO2 was lesser in in vitro vessels containing HT compared to control and in vitro vessels containing saponin, HT+DFM and Saponin+DFM. At 12 h, alpha and beta diversity of the microbial community between treatments differed (P<0.05). Across all samplesat 18 h of fermentation the genera Sharpea and Dialister were present, however Saponin+DFM and TCH+DFM treatments demonstrated the greatest relative abundance. While Sharpea contribute to rumen biohydrogenation, the presence of genus Dialister indicates association with energy harvesting bacteria and suggests that the inclusion of several of these feed additives allowed shifts of certain bacteria to occur.

Determination of tannin acyl hydrolase activity of Streptomyces mirabilis TBGS10 by measuring gallic acid using high-pressure liquid chromatography

Tannase is a hydrolytic enzyme known as tannin acyl hydrolase that acts on the ester bonds of hydrolyzable tannins and produces gallic acid. Tannase enzymes are obtained from various sources. Tannase from microbial sources, especially fungi like Aspergillus spp., has been used in the food, brewing, and pharmaceutical industries. Tannin acyl hydrolase activities of Actinomycetes are seldom reported, though they are known to produce a wide range of industrial enzymes. This communication deals with the tannase activity of a Streptomyces isolate from the shola forests of Munnar in the Western Ghats of Kerala. The isolate is identified as S. mirabilis TBGS10 based on morphological characters and 16s rDNA homology. The isolate showed promising tannase activity in plate assays, submerged, and solid-state fermentation conditions. High-pressure liquid chromatography determines the industrially important intermediate gallic acid produced through solid-state fermentation using cashew apple bagasse as substrate. The extract observed a gallic acid (GA) content of 142.624 g/mL with a retention time of 2.506 min. The tannase gene of the strain TBGS10 was PCR amplified using specific primers designed based on similar Streptomyces sequences available in the NCBI-GenBank

Solid-Phase Extraction at High pH as a Promising Tool for Targeted Isolation of Biologically Active Fractions of Humic Acids.

A search for novel sources of biologically active compounds is at the top of the agenda for biomedical technologies. Natural humic substances (HSs) contain a large variety of different chemotypes, such as condensed tannins, hydrolyzable tannins, terpenoids, lignins, etc. The goal of this work was to develop an efficient separation technique based on solid-phase extraction (SPE) for the isolation of narrow fractions of HS with higher biological activity compared to the initial material. We used lignite humic acid as the parent humic material, which showed moderate inhibition activity toward beta-lactamase TEM 1 and antioxidant activity. We applied two different SPE techniques: the first one was based on a gradient elution with water/methanol mixtures of the humic material sorbed at pH 2, and the second one implied separation by a difference in the pKa value by the use of sequential sorption of HS at pH from 8 to 3. SPE cartridges Bond Elute PPL (Agilent) were used in the fractionation experiments. The first and second techniques yielded 9 and 7 fractions, respectively. All fractions were characterized using high-resolution mass spectrometry and biological assays, including the determination of beta-lactamase (TEM 1) inhibition activity and antioxidant activity. The acidity-based separation technique demonstrated substantial advantages: it enabled the isolation of components, outcompeting the initial material at the first step of separation (sorption at pH 8). It showed moderate orthogonality in separation with regard to the polarity-based technique. Good perspectives are shown for developing a 2D separation scheme using a combination of polarity and acidity-based approaches to reduce structural heterogeneity of the narrow fractions of HS.

Phytochemical Investigation of Sumac (Rhus coriaria L.) Fruits from Different Sicilian Accessions.

Sumac, Rhus coriaria L., is employed as a natural preservative in the food sector, due to its rich content of antioxidant compounds, including hydrolysable tannins, phenolic acids, anthocyanins, and flavonoids. In this work, the phytochemical characterization of sumac fruits from five Sicilian accessions was performed to evaluate their potential as a food preservative for nutraceutical exploitation. Spectrophotometric tests and HPLC-MS/MS analyses were conducted to assess and compare the antioxidant power of the water extracts produced with the five sumac accessions. Principal component analysis was also carried out to better visualize the obtained results. Flavonoids and phenolic acids, namely isoquercitrin (20,342.82 mg/kg dry extract) and gallic acid (197,489.19 mg/kg dry extract), were more abundant in fruits from the population of San Biagio Platani, while the one from Giarratana was characterized by a higher content of anthocyanins such as cyanidin-3-glucoside (20,889.81 mg/kg dry extract). These two populations can be recognized as the most suitable settings for the implementation of sumac cultivation and the development of sumac-based products, especially for food and nutraceutical purposes.

Comparative studies of phytonutrients content and therapeutic potential of fruits parts of Dialium guineense Willd (Fabaceae) and Ziziphus mauritiana Lam. (Rhamnaceae) used in traditional pediatric in Burkina Faso

ABSTRACT Objective: The objective of the present study was to determine the phytonutrients content and selected biological activities of fruit parts of Dialium guineense commonly known as velvet tamarind and Ziziphus mauritiana called jujube tree used in the management of childhood diseases. Methodology and Results: Phytochemical components were determined by specific reagents and biological activities by antiradical and antibacterial test. The fruit pulp of Ziziphus mauritiana had the best antioxidant activity (IC50 = 0.27 mg/mL) due to its high contents ( p < 0.05 ) of total phenolic compounds (31.18 ± 0.06 mg GAE/g), flavonoids (0.73 ± 0.50 mg RE /g), hydrolysable tannins (11.37 ± 0.05 mg TAE/g) and condensed tannins (9.86 ± 0.30 mg CE/g). It also exhibited the highest contents of -carotene (72 ± 0.01 mg/100g) and macroelements Na (12 ± 0.00 mg/100g), K (605 ± 0.20 mg/100g). On the other hand, the fruit pulp of Dialium guineense had the best contents of vitamin C (36 ± 0.07 mg/100g), vitamin E (0.109 ± 0.07 mg/100g) and microelements (Fe, Zn) with 8.00 ± 0.01 mg/100g and 1.10 ± 0.02 mg/100g respectively. The fruits pulp of these two species showed antibacterial activity against E. coli with the best minimum inhibitory concentration (MIC) of 3.12 mg/mL Conclusions and application of results: Recommendations of enhancement and promotion of these two fruit species could constitute important sources for the development of new nutraceuticals and phytomedicines in the prevention and treatment of childhood diseases. Keywords: Childhood diseases, Dialium guineense, Ziziphus mauritiana, Nutraceuticals.

Anti-Culex pipiens activity of different pomegranate cultivars and determination of their bioactive compounds using LC-MS profiling.

Pomegranate (Punica granatum L.) peels are rich in various bioactive compounds. Characterization of these compounds is crucial for the utilization of peel waste in industrial processing. The study aimed (1) to establish and compare the metabolic profiles of the peel of seven pomegranate cultivars and (2) to identify bioactive compounds contributing to the larvicidal activity against the third instar larvae of Culex pipiens. UPLC-ESI-MS/MS was utilized to analyze peel methanol extracts of different pomegranate cultivars. The larvicidal activity was determined by calculating the larval mortality among the third instar larvae of C. pipiens. Multivariate data analysis was conducted to identify the metabolites that exhibited a larvicidal effect. A total of 24 metabolites, including hydrolyzable tannins, flavonoids, and alkaloids, were tentatively identified in both negative and positive ionization modes. The extract of cultivar 'Black' exhibited the most potent larvicidal effect with LC50 values of 185.15, 156.84, and 138.12ppm/mL after 24, 48, and 72 h of treatment, respectively. By applying chemometric techniques, the larvicidal activity could be directly correlated to the bioactive compounds punicalagin, quercetin-O-rhamnoside, quercetin-O-pentoside, and galloyl-HHDP-glucose. The present study implemented UPLC-ESI-MS/MS and chemometric techniques as potential tools for metabolomics analysis and differentiation between peels of different pomegranate cultivars. In addition, cultivar 'Black' extract could be a promising natural insecticide against mosquitoes since it is rich in bioactive compounds with larvicidal activity.

Phytochemical analysis, antioxidant, and antimicrobial activities of Jordanian Pomegranate peels.

Pomegranate (Punica granatum) peels have shown numerous health benefits such as antioxidant, anti-inflammatory, and antimicrobial activities. These health activities are owed to the unique phytochemical components present in pomegranate peels. Variations in the pomegranate cultivar, geographical region, and extraction methods significantly affect the phytochemical composition and concentrations of pomegranate fruits and their peels, hence their health outcomes. Therefore, this study aimed to examine the phytochemical contents of pomegranate peels of Jordanian origin and their antioxidant and antimicrobial activities. Among the 6 extracts of pomegranate peels tested, the ethanol extract exhibited the highest total phenolic content (TPC = 297.70 ± 1.73 mg GAE/g DW), highest total flavonoids content (TFC = 116.08 ± 3.46 mg RE/g DW), highest hydrolyzable tannins (HT) contents (688.50 ± 3.54 mg TE/g DW). Whereas the highest condensed tannins (CT) content was found in both the ethanol (13.87 ± 0.58 mg CE/g DW) and methanol (13.84 ± 0.55 mg CE/g DW) extracts. For the antioxidant activities, the water extract of pomegranate peels displayed the highest inhibitory effect on DPPH radicals (9.43 ± 0.06 μmole TE/g DW), while for the ABTS+ assay the methanol and ethanol extracts exhibited the highest activities of 11.09 ± 0.02 and 11.09 ± 0.06 μmole TE/g DW, respectively. For the FRAP assay, the aqueous methanol extract exhibited the highest reducing activity (1.60 ± 0.09 mmole Fe (II)/g DW). As for the antimicrobial activities of various extracts of pomegranate peels, the highest antimicrobial activity against Micrococcus luteus was achieved by the ethanol extract (MIC = 6.25 mg/mL), whereas the lowest antimicrobial activity was observed against Candida krusei using the methanol extract (MIC = 100 mg/mL). These results indicate that pomegranate peels of Jordanian origin are rich in phytochemical content and exhibited strong antioxidant and antimicrobial activities making these agroindustrial by-products potential candidates for various medical applications and possible safe sources for important bioactive components.

Influence of eco-friendly dispersants on the properties of a lateritic soil-based mortar

Tropical regions like French Guiana need local building materials to cope with high population growth and the high cost of imported cementitious materials. Poured earthen construction could represent a local, cost-effective, and ecologically friendly alternative. To ensure good workability and facilitate pouring, the use of dispersants to deflocculate clay particles is an effective strategy that reduces water demand and increases the material's density and strength. Natural organic dispersants can replace industrial ones while reducing costs and carbon footprint. It is currently unknown how organic dispersants could improve the workability, physical and mechanical properties of the iron-rich lateritic soils present in French Guiana. Here, different potential dispersants were evaluated at constant water content on a lateritic soil-based mortar: citric acid, sodium carbonate, tannins, tannins+sodium hydroxide (tannins+NaOH), tannins+sodium carbonate (tannins+Na2CO3). These dispersants were compared to industrial sodium hexametaphosphate (NaHMP). Three types of tannins were tested: hydrolyzable tannins from oak and chestnut, and condensed tannins from acacia. This study shows that all formulations improved workability and mechanical strength but only tannins+NaOH or Na2CO3 had a strong dispersant effect comparable to NaHMP. Furthermore, tannins+NaOH or Na2CO3 decreased the mortar's density without impacting strength, which may result from reactions between the soil's iron oxides and tannins, as observed by infrared spectroscopy (FTIR). Altogether, these results show that the addition of organic dispersant is an appropriate strategy to improve the fresh and hardened properties of lateritic soils. Particularly, tannins combined with sodium carbonate may represent an eco-friendly dispersant for poured earth in regions with iron-rich lateritic soils.

Tannase production using green biotechnology and its applications: A review

An enormous increase in the use of enzymes in a wide range of industrial biological conversion technologies has been made possible by the explosion of green biotechnology. Industrial enzymes' huge biocatalytic potential gives them an advantage over chemical technology in terms of security, reusability, and greater process regulation. One such enzyme with enormous potential for converting hydrolyzable tannins into gallic acid is tannase. Tannins mostly accumulate in plant components including leaves, fruits, roots as well as bark. Additionally, harmful tannery effluents from a variety of tanneries contain high concentrations of tannins in the chemical form of tannic acid. Tannase is primarily used to degrade the tannins that mostly exist in harmful tannery effluents, offering a comparatively much more affordable method for their biodegradation. This review discusses the mechanism of action of tannases, with an emphasis on substrates and production, biochemical characterization, and its applications.

Valorization of Prickly Pear Peel Residues (Opuntia ficus-indica) Using Solid-State Fermentation.

Prickly pear peel (Opuntia ficus-indica) residues can be used as a substrate in solid-state fermentation to obtain bioactive compounds. The kinetic growth of some Aspergillus strains was evaluated. A Box-Hunter and Hunter design to evaluate the independent factors was used. These factors were temperature (°C), inoculum (spores/g), humidity (%), pH, NaNO3 (g/L), MgSO4 (g/L), KCl (g/L), and KH2PO4 (g/L). The response factors were the amount of hydrolyzable and condensed tannins. The antioxidant and antimicrobial activity of fermentation extracts was evaluated. Aspergillus niger strains GH1 and HT3 were the best for accumulating tannins. The humidity, inoculum, and temperature affect the release of hydrolyzable and condensed tannins. Treatment 13 (low values for temperature, inoculum, NaNO3, MgSO4; and high values for humidity, pH, KCl, KH2PO4) resulted in 32.9 mg/g of condensed tannins being obtained; while treatment 16 (high values for all the factors evaluated) resulted in 3.5 mg/g of hydrolyzable tannins being obtained. In addition, the fermented extracts showed higher antioxidant activity compared to the unfermented extracts. Treatments 13 and 16 showed low inhibition of E. coli, Alternaria sp., and Botrytis spp. The solid-state fermentation process involving prickly pear peel residues favors the accumulation of condensed and hydrolyzable tannins, with antioxidant and antifungal activity.

Application of response surface methodology in the characterization and sensorial analysis of Anacardium humile liqueur

Anacardium humile, known as “cajuzinho-do-cerrado,” is a native Brazilian Cerrado fruit similar to the cashew but less studied, particularly its phenolic compound content and profile. This study used a factorial design to examine how A. humile concentration and maturation time affect the liqueur's physicochemical attributes, phenolic content, antioxidant capacity, and sensory appeal. The liqueurs exhibited low antioxidant capacities as determined by the 2,2-diphenyl-1-picrylhydrazyl radical scavenging capacity assay (DPPH), ferric ion reducing antioxidant power (FRAP), and 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid). This is likely attributable to a reduction in phenolic compounds within the liqueurs compared to A. humile. The phenolic compound profile of A. humile showed 11 compounds, comprising 5 flavonols, 3 hydroxycinnamic acids, and 2 hydrolyzable tannins. Treatment 8 (500 g of A. humile and 17 days of maturation) showed the highest value for phenolic compound content. However, with regard to sensory analysis, this was the darkest liqueur and the least accepted test, while Treatment 5 (147.5 g of A. humile and 10 days of maturation) was one of the most accepted and chosen as the best liqueur due to the highest antioxidant capacity by FRAP and DPPH methods. The fusion of technology and factorial design proved efficacious in formulating functional liqueurs.

Development and Acceptability of Cookie Product Made from Marang (Artocarpus Odoratissimus) Seed Flour

Marang, known for its distinctive flavor and texture, is a tropical fruit hailing from Southeast Asia, featuring sweet, custard-like flesh and a fragrant scent. This study aimed to create Marang seed (Artocarpus odoratissimus) flour and cookies, establishing the optimal ingredient ratios to deliver a delightful flavor. Employing both quantitative and qualitative approaches, this research enlisted consumer respondents and food professionals to assess sensory qualities using the DOST’s 9-point hedonic scale. Statistical analysis, including mean and standard deviation, was employed to evaluate cookie acceptability based on qualitative criteria encompassing appearance, aroma, taste, and texture. Phyto-chemical analysis revealed the presence of alkaloids, naturally occurring substances with potential pharmacological effects, in Marang extract. Quaternary bases and amin e oxides, organic compounds featuring positively charged nitrogen atoms, were identified, alongside steroids with diverse biological functions. The grading system denoted the relative concentration of alkaloids. While 2-deoxysugars were detected, unsaturated steroids were absent. Flavonoids, known for their antioxidant properties, and saponins, foaming agents with potential health benefits, were also present. Tannins, recognized for their astringency, appeared with different colors indicating condensed and hydrolyzable tannins. Marang seed flour exhibited a chemical composition with 18.2% oil content, 16% moisture content, 10.7% crude protein content, and notable levels of manganese and copper at 31.8 ppm and 7.5 ppm, respectively, along with 161 ppm of phosphorus. The overall acceptability of the developed cookie was moderately liked, with a grand mean rating of 7.76. Taste held the greatest influence on overall acceptability, followed by texture, appearance, and aroma. This study recommends further investigation into the pharmacological effects and potential health benefits of Marang extract components, potentially leading to pharmaceutical or functional product development. Utilizing Marang seed flour in various food products can leverage its nutritional value and meet potential commercial demand. Improving and standardizing the Marang cookie recipe for uniform quality and flavor is advisable. The development of packaging and branding to highlight the unique qualities and health benefits of Marang cookies can cater to health-conscious consumers.

340 Ruminal Protozoa Viability Under Diverse Additives Using Trypan Blue Exclusion Method as an Alternative Counting Methodology

Abstract Beef cattle are a major contributor to methane (CH4) emissions. Enteric CH4 is produced by archaea, mostly hosted by ruminal protozoa, thus, changes in protozoa counts have been associated with CH4 production. Therefore, reliable ruminal protozoa count (PROT) is essential to evaluate CH4-mitigation strategies. Typically, PROT are done by fixation with methyl-green-formalin-saline solution (MFS), which reveals ciliate numbers but not ruminal protozoa viability (VIAB). The Trypan Blue Exclusion Method (TBEM) involves an intracellular probe that dyes the cytoplasm of apoptotic eukaryotic cells with membrane damage, but few studies report its use for PROT. Therefore, our objectives were to establish culture conditions to use the TBEM for PROT and VIAB under different diets (Exp. 1) and feed additives. In Exp. 1 ruminal fluid was collected from 4 Angus-crossbred ruminally cannulated steers receiving a high-grain diet (NOYE) alone or supplemented with 28 g /d of a product of yeast fermentation (YEAST). Filtered and diluted ruminal fluid was incubated (39ºC, 95% CO2) with (SUB) or without (NOSUB) the substrate diet in three types of containers; 125 mL bottles (BOT), Erlenmeyer (ERL), or 16 × 125 mm Hungate tubes (TUB). After 8, 12, 24, and 48 h PROT and VIAB were determined using the TBEM, gas production, and CH4 concentration were measured. Exp. 2 involved in vitro batch culture incubations with a silage-based substrate (CON), plus a 50:50% mix of condensed and hydrolysable tannins (TAN), cashew nutshell extract (CNSE), or citral (CIT) at 0.3, 0.02, and 0.2% of substrate DM, respectively. At 0 and 48 h PROT, VIAB, volatile fatty acids (VFA), CH4, in vitro organic matter digestibility (IVOMD), and gas were determined. In Exp-1, PROT and VIAB were greater in TUB (P = 0.01) at 24 h and in BOT at 48 h (P < 0.01), particularly with SUB (P < 0.01). Supplementation of YEAST reduced VIAB in BOT and TUB compared with ERL (P < 0.01). In Exp. 2, TAN (P < 0.01) and CIT (p 0.05) reduced PROT and VIAB at 24 and 48 h (P < 0.01). Additionally, TAN reduced gas production (P < 0.01), and IVOMD (P < 0.01) coinciding with previous studies reporting protozoa reduction. Protozoa/mL were similar to previous reports with MFS in cows (1.5 × 104) and steers (3.7 × 103). In both experiments, PROT and VIAB reductions by YEAST, TAN and CIT coincided with a reduction of CH4, gas, and IVOMD. Differences in PROT with some studies using MFS might be associated with losses of small Entodinium during initial filtrations. When cultured in BOT with substrate, TBEM is a viable, simple, rapid, and cost-effective methodology to evaluate PROT and VIAB after a nutritional challenge. The ability to assess membrane disruptions associated with protozoa apoptosis in the TBEM is one its greatest advantages.