Acetic Research Articles

Impacts of ohmic-heating on the surface characteristics, membrane integrity, morphology, and chemical compounds of Bifidobacterium animalis paraprobiotic

The aim of this paper was to obtain paraprobiotic from Bifidobacterium animalis subsp. lactis Bb-12 (BB) with a promising ohmic-heating (OH) condition through response surface modeling, based on the flow cytometry and spectrophotometry analyses. Some properties of the obtained BB paraprobiotic (BBP) were studied, including the cell surface hydrophobicity (CSH), cell auto- and co-aggregation (CAA and CCA), and membrane integrity (MI). Among fitted models, the linear model was more accurate for co-aggregation (R2-pred 85.77%), and the quadratic model for CSH (R2-pred: 84.17%), auto-aggregation (R2-pred=87.25%), MI (R2-pred: 83.65%). The electric field (EF) had the most destructive effect on the MI of BBP. However, OH temperature had the most enhancing effect on the surface characteristics. The surface characteristics of BBP (CSH: 44.82, CAA: 38.05, and CCA: 40.79) obtained by the OH process at optimum condition was remarkably more than BB without OH treatment (CSH: 34.21, CAA: 27.18, and CCA: 28.5). The confirming analysis regarding the MI changes was performed successfully by scanning electron microscopy analysis. Lactic and acetic acids were the most amounts of characterized metabolites in BBP and BB. The OH condition, including EF of 8 V/cm, OH temperature of 88˚C, BB population of 8 (log CFU/mL), and OH time of 3 min, were considered optimum condition. At this optimum OH condition, the responses for CSH, CAA, CCA, and MI were observed 44.82, 38.05, 40.79, and 94.82 %, respectively.

An efficient synthesis, structural analysis, and computational studies of benzothiazole derivatives activated by formic acid under solvent-free conditions

In the present work, formic acid was used as a low-cost and eco-friendly catalyst to synthesize 2-(2-hydroxyphenyl)benzothiazole (HB) through a cycloaddition process in a solvent-free reaction medium. Imine molecule was synthesized with the help of acetic acid (B). The ring closure and formation of the thiazole moiety give acetic acid distinctiveness as a catalyst. The cyclization of fused HB failed without an acid catalyst. The chemical structures were characterized using microanalysis techniques such as IR, 1H-NMR, and 13C-NMR. The obtained FT-IR spectra were compared to theoretical data using the B3LYP/6-31G* level of theory. The electronic characteristics of molecules can be predicted utilizing wave function-dependent features, such as Average localized ionization energy (ALIE), Localized orbital locator (LOL), Electron localized function (ELF), Reduced density gradient (RDG), and Noncovalent interactions NCI studies, which have provided a wealth of knowledge. The molecular docking study was investigated to explore binding energy and interaction type. Docking analysis displayed binding energies (kcal/mol) of HB with values of -8.29, -6.80, -6.68, -7.43, and -6.69 of acetylcholinesterase (AChE) and butylcholinesterase (BuChE)), SARS-CoV-2 main protease (Mpro), human immunodeficiency virus reverse transcription (HIV RT)) and cytochrome (P450), respectively, which was the most active compound against all receptors. The dramatic improvement of the proposed approach over the existing methods can be used to derive an efficient and successful route for the synthesis of HB. This process provides a fresh and high-yield method for the synthesis of benzimidazoles.

Drought priming improves tolerance of Alhagi sparsifolia to subsequent drought: A coordinated interplay of phytohormones, osmolytes, and antioxidant potential

Perennial trees are often stressed by drought more than once during their life cycle. Our study exposed three-month-old Alhagisparsifolia, with (drought-primed) or without (nonprimed) prior drought stress to subsequent drought for two months, aiming to reveal whether pre-exposure to drought could enhance seedling resistance to subsequent drought and investigated possible underlying mechanisms. Root biomass, leaf relative water content, chlorophyll a, and carotenoids were significantly higher in drought-primed than nonprimed seedlings. They also had reduced concentrations of malondialdehyde, hydrogen peroxide (H2O2), and superoxide anions (O2•−), indicating relief from oxidative stress. This relief was associated with a coordinated upregulation of enzymes scavenging O2•− and H2O2, particularly superoxide dismutase (SOD) and catalase (CAT), and the maintenance of the ascorbate-glutathione (AsA-GSH) redox pool and enzymatic activities (ascorbate peroxidase, mono- and dehydroascorbate reductase, and glutathione reductase), leading to the better regulation of reactive oxygen species. The failure of nonprimed seedlings to upregulate the SOD, CAT, and AsA-GSH cycles nevertheless made the seedlings susceptible to oxidative stress. The increased levels of strigolactones, jasmonic acid, and abscisic acid in drought-primed seedlings reveal their roles in subsequent stress. They also displayed higher gibberellic acid and indole acetic acid. A principal component analysis showed that the seedlings responded differently to drought if they had previously suffered a drought, mainly due to a higher capacity for pigment protection, oxidative scavenging, osmolytes, and anti-stress hormones. Our study provides insights into the benefits of stress memory induced in seedlings by early drought priming as a strategy for overcoming subsequent stress.

Integrated multiple-omics reveals the regulatory mechanism underlying the effects of homologous Bacillus tequilensis (GCB-3) on growth performance of grass carp (Ctenopharyngodon idellus)

The purpose of this study was to investigate the regulatory mechanism of Bacillus tequilensis (GCB-3) in improving the growth performance of grass carp by microbiome and targeted metabolomics analysis. Fish were fed with different amounts of GCB-3 including B0 (0 CFU/g), B6 (1×106 CFU/g), B7 (1×107 CFU/g), B8 (1×108 CFU/g), and B9 (1×109 CFU/g) for 8 weeks. The results indicate that with the increase of dietary GCB-3 concentration, grass carp showed a significant linear trend in growth performance, and appetite factors showed a significant linear and quadratic trend, the growth performance and appetite-promoting factors in the B8 group reach their maximum value in the B8 group. Microbiome analysis found that Proteobacteria and Actinobacteria were the most abundant phylum in the gut microbiota of grass carp. The functional potential predicted that GCB-3 affected the synthesis and degradation of carbohydrates, amino acids, fatty acids and lipids. It found that GCB-3 also affected the tricarboxylic acid (TCA) cycle VII (acetate-producers) pathway. The results of targeted metabolomics showed a significant linear and quadratic increase in intestinal acetate with GCB-3 concentration and a maximum in the B8 group. The results of the correlation analysis showed a significant positive correlation between Actinobacteria abundance and acetic acid content, and also between TCA cycle VII (acetate-producers) pathway abundance and Actinobacteria abundance and acetic acid content. In B8 group, grass carp showed a faster regulation speed of blood glucose levels after glucose injection, and the GLP-1 content remained stable. The activities of glycolytic enzymes, lipolytic enzymes, protein metabolism enzymes, and their related mRNA expression levels in grass carp showed a linear or quadratic increasing trend. However, the expression levels of gluconeogenic enzymes and fatty acid synthesis enzyme related mRNA showed an opposite trend. In summary, the addition of 1×108 CFU/g GCB-3 in grass carp feed can improve the abundance of dominant bacterial phyla in the intestine, especially Actinobacteria, increase the concentration of intestinal acetate, provide substrate for the TCA cycle VII (acetate-producers) pathway, thereby regulating the nutrient metabolism of grass carp and ultimately promoting growth.

Revealing the microbial diversity and volatile flavor formation in finger citron kombucha by metagenomic and GC-MS analysis

Kombucha is a kind of health drink formed by the metabolism of Symbiotic Culture of Bacteria and Yeast (SCOBY) composed of yeast, acetic acid bacteria and lactic acid bacteria, which is popular among researchers and fond of consumers because of its antioxidant and hypoglycemic effects. In this study, the finger citron kombucha with different fermentation time (2, 4, 6, 8 day) were analyzed by headspace solid phase micro-extraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) and metagenomic sequencing. The results showed that a total of 76 volatiles were identified, including 15 key volatiles, such as linalool, geraniol, α-terpinene, and limonene, which had a great influence on the flavor of kombucha. Metagenomic analysis showed that unclassified-g-Gluconobacter, Komagataeibacter saccharivorans, Gluconobacter-sp.-R71656 and other microorganisms were the dominant species in finger citron kombucha. Through the correlation heat map analysis of volatiles and microbial composition, it was found that Gluconobacter had a positive effect on the formation of most volatiles of kombucha. In addition, Novacetimonas hansenii, Brettanomyces bruxellensis and Wickerhamiella sorbophila were also closely related to the flavor formation of kombucha. This study provided a theoretical basis for finding the key aroma-producing microorganisms in the fermentation process of finger citron kombucha and the subsequent construction of SCOBY.

Associations of per- and polyfluoroalkyl substances with maternal early second trimester sex-steroid hormones

Background/AimsPregnant women are exposed to persistent environmental contaminants, including per- and polyfluoroalkyl substances (PFAS) that disrupt thyroid function. However, it is unclear if PFAS alter maternal sex-steroid hormone levels, which support pregnancy health and fetal development. MethodsIn Illinois women with relatively high socioeconomic status (n = 460), we quantified perfluorononanoic (PFNA), perfluorooctane sulfonic (PFOS), perfluorooctanoic (PFOA), methyl-perfluorooctane sulfonamide acetic acid, perfluorohexanesulphonic (PFHxS), perfluorodecanoic (PFDeA), and perfluoroundecanoic (PFUdA) acid concentrations in fasting serum samples at median 17 weeks gestation, along with plasma progesterone, testosterone, and estradiol. We evaluated covariate-adjusted associations of ln-transformed hormones with each ln-transformed PFAS individually using linear regression and with the PFAS mixture using quantile-based g-computation (QGComp). ResultsInterquartile range (IQR) increases in PFOS were associated with higher progesterone (%Δ 3.0; 95%CI: −0.6, 6.6) and estradiol (%Δ: 8.1; 95%CI: 2.2, 14.4) levels. Additionally, PFHxS was positively associated with testosterone (%Δ: 10.2; 95%CI: 4.0, 16.7), whereas both PFDeA and PFUdA were inversely associated with testosterone (%Δ: −5.7; 95%CI: −10.3, −0.8, and %Δ: −4.1; 95%CI: −7.6, −0.4, respectively). The IQR-standardized PFAS mixture was not associated with progesterone (%Δ: 1.6; 95%CI: −5.8, 9.2), due equal partial positive (%Δ: 9.2; driven by PFOA) and negative (%Δ: −7.4; driven by PFOS) mixture associations. Similarly, the mixture was not associated with testosterone (%Δ: 5.3; 95%CI: −9.0, 20.1), due to similar partial positive (%Δ: 23.6; driven by PFHxS) and negative (%Δ: −17.4; driven by PFDeA) mixture associations. However, we observed a slightly stronger partial positive (%Δ: 25.6; driven by PFOS and PFUdA) than negative (%Δ: −16.3; driven by PFOA) association resulting in an overall non-significant positive trend between the mixture and estradiol (%Δ: 8.5; 95%CI: −3.7, 20.9). ConclusionPFAS mixture modeled using QGComp was not associated with maternal sex-steroid hormones due to potential opposing effects of certain PFAS. Additional prospective studies could corroborate these findings.

Preparation and evaluation of Diclofenac & Aceclofenac pain relieving gel

It is phenyl acetic acid derivative developed as anti-inflammatory agent. It has analgesic anti-inflammatory antipyretic like actions like other NSAIDS. . It is recommended in long term treatment of rheumatoid arthritis, osteoarthritis and enclosing spondylitis. It is also useful acute miscue skeletal disorder post operative pain and dysmenorrhoea Diclofenac sodium gel were developed in seven different formulations) by employing different grades of polymers such as Hpmck4m and Crbopol940. There the various Diclofenac gels are available in market. But the propose gel is formulated with two key ingredients oleoresin, and l linseed oil contribute to anti-inflammatory effect. The formulations were evaluated for various physical parameters, pH spredability, drug release excludability studies drug released mechanisms. Formulation showed maximum drug release of 8 hours and maximum drug. Finally the gel formulations found to be economical and may overcome the draw backs associated with the drug during its absorption. The purpose of the study is to prepare the combination of diclofenac and aceclofenac pain relieving gel. This gel combined form of gel is multifunctional and getting rapid onset of action. And that is to get instant relieve from pain as comparison of plain aceclofenac tablet and plain Diclofenac gel. This formulation is also beneficial for diabetic patient to get cure fast of skin injury and buildup of skin irritation and growth or new epidermis layer of skin in the body.

Evaluation of anti- inflammatory and analgesic activity of Bhasmas in experimental animal models

Acute toxicity studies of both the bhasmas showed no mortality at 2000 mg/kg.The Scanning electron microscopy of both Tamra and Lauha bhasma indicated the nanomaterial characteristics which justifies claim of Ayurvedic bhasmas as ancient nanomaterials. Tamra and Lauha bhasma were found to be effective as anti-inflammatory agent in acute models of inflammation. It inhibits all phases of inflammation induced by carrageenan. Tamra and Lauha bhasma were found to be effective as anti-inflammatory agent in chronic model of inflammation. It significantly inhibits wet as well as dry weight of cotton pellet granuloma at highest doses only. Analgesic activity of both Tamra and Lauha bhasma in hot plate and tail immersion models were found to be significant at 120-240 mins. Analgesic activity of both Tamra and Lauha bhasma in acetic acid models were found to be non-significant in nature, while found to be significant at late phase in formalin induced model. Tamra and Lauha bhasma also were found to have significant anti-arthritic activity in animal study.The whole pharmacological investigations were indicated that activity was only significant at therapeutic and more than therapeutic dose but nonsignificant at dose lower than therapeutic doses. The obtained results indicates the anti-inflammatory and analgesic potential of both these bhasmas probably by inhibiting inflammatory mediators.

Aspergillus awamori: potential antioxidant, anti-inflammatory, and anti-apoptotic activities in acetic acid-induced ulcerative colitis in rats.

Ulcerative colitis (UC) is a chronic colonic inflammation with a significant health hazard. Aspergillus awamori (A. awamori) is a microorganism with various bioactive compounds with natural antioxidant and anti-inflammatory properties. The present work aimed to elucidate the protective and therapeutic effects of varying concentrations of A. awamori against acetic acid (AA)-induced ulcerative colitis (UC) in rats. Nine groups of albino male rats were established: a control negative group (G1), a control positive group (G2,AA), and preventive protocol groups (including G3A, G4A, and G5A) that received 100mg, 50mg, and 25mg/kg b.w, respectively, of A. awamori orally and daily from the 1st day of the experiment and for 7 consecutive days. Then, they were subjected to one dose of AA intrarectally on day 8th. G3B, G4B, and G5B were termed as curative protocol groups that received one dose of AA on day 8th and then administered 100mg, 50mg, and 25mg/kg b.w. of A. awamori, respectively, on day 9th and continued receiving these doses daily until day 16th. Rats in the AA group exhibited marked histopathological alterations of the distal colon, with an exaggeration of the DAI. In addition, a remarkable increase in oxidative stress was represented by the elevation of MDA and NO levels with a decline in SOD and GPx activities. In addition, upregulation of TNF-α, IL-6, and IL-1β mRNA expressions and downregulation of Muc2 and Nrf2 levels were detected. Unambiguously, a remarkable anti-inflammatory effect was noticed either in A. awamori prevented or treated groups expounded by reducing and regulating TNF-α, IL-6, and IL-1β with improved pathological lesion scoring. The Muc2, Nrf2, and bcl-2 gene levels were upregulated and restored also. In summary, the findings in this work reveal that A. awamori supplementation successfully alleviated the UC induced by AA, which had a better effect when administered before colitis induction.

The use of thioxopyrimidine derivatives for the regulation of vegetative growth of wheat

The work is devoted to the screening of new effective wheat plant growth regulators based on synthetic compounds, thioxopyrimidine derivatives. The wheat (Triticum aestivum L.) variety Oksamit Mironivskyi was used as the object of the study. Morphometric parameters (average length of shoots and roots (mm), and average biomass of 10 plants (g)) of 4-week-old wheat plants were measured. Treatment of wheat plants with thioxopyrimidine derivatives at a concentration of 10-6 M contributed to the growth and development of shoots and roots compared to control wheat plants treated with distilled water. The regulatory effect of thioxopyrimidine derivatives on the vegetative growth of wheat plants was similar to the regulatory effect of auxin IAA (1H-indol-3-yl)acetic acid) or synthetic compounds, derivatives of sodium and potassium salts of 6-methyl-2-mercapto-4-hydroxypyrimidine (Methyur, Kamethur) and N-oxide-2,6-dimethylpyridine (Ivin) used in the same concentration of 10-6 M. The correlation between the growth regulatory effect and the chemical structure of synthetic compounds, thioxopyrimidine derivatives was established. The selected most active synthetic compounds, thioxopyrimidine derivatives, are proposed to be used to improve the vegetative growth of wheat (T. aestivum L.) variety Oksamit Mironivskyi.

Bifidobacterium longum S3 alleviates loperamide-induced constipation by modulating intestinal acetic acid and stearic acid levels in mice.

Constipation is a major gastrointestinal (GI) symptom worldwide, with diverse causes of formation, and requires effective and safe therapeutic measures. In the present study, we used loperamide hydrochloride to establish a constipation model and assessed the effect of Bifidobacterium on constipation and its possible mechanism of relief. The results showed that B. longum S3 exerted a constipation-relieving effect primarily by improving the gut microbiota, enriching genera including Lactobacillus, Alistipes, and Ruminococcaceae UCG-007, and decreasing the bacteria Lachnospiraceae NK4B4 group. These changes may thereby increase acetic acid and stearic acid (C18:0) levels, which significantly increase the expression levels of ZO-1 and MUC-2, repair intestinal barrier damage and reduce inflammation (IL-6). Furthermore, it also inhibited oxidative stress levels (SOD and CAT), decreased the expression of water channel proteins (AQP4 and AQP8), significantly elevated the Gas, 5-HT, PGE2, and Ach levels, and reduced nNOS and VIP levels to improve the intestinal luminal transit time and fecal water content. Collectively, these changes resulted in the alleviation of constipation.

Identification of volatile and non-volatile migrants released during sous vide cooking by UPLC-IMS-QTOF and DI-SPME-GC-MS using a design of experiments approach

The rise of new culinary advances combined with science has made it possible to find new optimum methods of cooking, such as vacuum cooking (Sous Vide), that allow achieving pleasant textures, preserving the genuine nature of the food and also its original flavour. Sous Vide involves the direct contact of plastic materials with food during cooking, causing the possible release of chemical compounds that may alter the properties of the food and pose a risk to human health. In this work, the migration kinetics that takes place during this cooking process have been estimated thanks to a design of experiments (DoE). DoE has been applied to 4 samples of vacuum cooking bags containing 3 food simulants (ethanol 10%, acetic acid 3% and isooctane) under typical cooking conditions of the culinary technique. The study also allowed the identification of the main volatile and non-volatile migrants that can be transferred from this particular packaging. Twenty-nine non-volatile species were identified by ultra-high performance liquid chromatography coupled to a quadrupole-time-of-flight ion-mobility separation mass spectrometer (UPLC-IMS-QTOF) and sixty-four volatile species were identified by solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). Furthermore, a comprehensive literature analysis was conducted to explore the applications, origins, and potential toxicity of these compounds.

Oral Administration Properties Evaluation of Three Milk-derived Extracellular Vesicles Based on Ultracentrifugation Extraction Methods.

Milk-derived extracellular vesicles (M-EVs) are low-cost, can be prepared in large quantities, and can cross the gastrointestinal barrier for oral administration. However, the composition of milk is complex, and M-EVs obtained by different extraction methods may affect their oral delivery. Based on this, we propose a new method for extracting M-EVs based on cryogenic freezing treatment (Cryo-M-EVs) and compare this method with the previously reported acetic acid treatment (Acid-M-EVs) method and the conventional ultracentrifugation method (Ulltr-M-EVs). The new method simplifies the pretreatment step and achieves 25-fold and 2-fold higher yields than Acid-M-EVs and Ulltr-M-EVs. And it was interesting to note that Cryo-M-EVs and Acid-M-EVs had higher cellular uptake efficiency, and Cryo-M-EVs presented the best transepithelial transport effect. After oral administration of the three M-EVs extracted by three methods in mice, Cryo-M-EVs effectively successfully crossed the gastrointestinal barrier and achieved hepatic accumulation, whereas Acid-M-EVs and Ultr-M-EVs mostly resided in the intestine. The M-EVs obtained by the three extraction methods showed a favorable safety profile at the cellular as well as animal level. Therefore, when M-EVs obtained by different extraction methods are used for oral drug delivery, we can utilize their accumulation properties at different sites to better deal with different diseases. This article is protected by copyright. All rights reserved.

In Vivo and In Silico Molecular Modeling Studies of Newly Synthesized Pyrazoles for Their Anti-Inflammatory and Analgesic Properties

Background: Pyrazole derivatives have gained attention as potential leads for developing anti-inflammatory and analgesic drugs. Chemical modifications of pyrazole structures offer a strategy to enhance their therapeutic properties, while minimizing undesirable effects. Objective: This study aimed to synthesize novel pyrazole derivatives through chemical modifications and assess their anti-inflammatory and analgesic activities. Methods: A series of pyrazole variants were synthesized by reacting various acetophenone derivatives and diethyl oxalate in basic conditions, followed by treatment with glacial acetic acid and hydrazine hydrate. The resulting compounds were further methylated and acetylated. The synthesized compounds were characterized using elemental analysis, 1H NMR, IR, and MASS spectroscopy. The anti-inflammatory activity was evaluated using the rat paw edema technique induced by carrageenan, while the analgesic activity was assessed using the writhing reflux method. Results: Among the newly synthesized pyrazole derivatives, compounds 3b, 3e and 3g′ demonstrated significant anti-inflammatory activity with reduced ulcerogenic potential. Compounds 3d, 3b′ and 3d′ exhibited notable analgesic activity compared to the standard drug. The chemical modifications introduced in these derivatives were confirmed through spectroscopic analyses. Conclusion: The findings of this study highlight the potential of pyrazole derivatives as promising candidates for anti-inflammatory and analgesic drug development. The structural modifications performed in this research improved the therapeutic activities of the compounds, while addressing safety concerns. Further investigations are warranted to elucidate the underlying mechanisms of action and optimize the pharmacological profiles of these novel pyrazole derivatives.

Effect of Recycled Concrete Aggregate Addition on the Asphalt Mixtures Performance: ITZ Area, Microstructure, and Chemical Analysis Perspectives

The importance of environmental consciousness and sustainability is increasing among transportation governing bodies worldwide. Many government bodies are concerned with maximizing the usage of recycled substances in road construction. Therefore, assessing the effect of recycled materials consumption is essential, mainly when designing new ‘green’ pavement types. The primary objective of this study is to investigate the impact of different treatments on improving the interfacial transition zone (ITZ) of coarse recycled concrete aggregate (CRCA) and its application in asphalt mixes. Such an aim is accomplished by enhancing its physical and mechanical characteristics, as well as its microstructure. The surface morphology, chemical composition, and intermix phases of the ITZ area and calcium silicate hydrate (CSH) compounds for CRCA were evaluated using scanning electron microscopy (SEM), an energy-dispersive X-ray analyzer (EDAX), and X-ray diffraction analysis (XRD). The performance of asphalt mixtures that included treated and untreated CRCA was also examined using different tests. It was found that heat treatment is an effective technique for enhancing the ITZ. However, cracks were seen in the mortar of CRCA when exposed to high temperatures (500 °C), which adversely affects the characteristics of the mortar. Acid treatment appeared to be an effective approach for improving the ITZ area. Nevertheless, the treatment that used acetic acid, a weak acid, was more effective than HCl acid, a strong acid. The outcomes revealed that the ITZ microstructure is significantly enhanced under different treatment types; however, microstructure improvements mainly included increased surface homogeneity and CSH compounds and a reduced Ca/Si ratio. It was also found that the asphalt mixtures with different proportions of untreated CRCA exhibited enhanced resistance to rutting. Furthermore, their tensile strength ratio (TSR) values were above the minimal level requirements. Moreover, the asphalt mixture with 30% CRCA, which was treated with various treatment methods, demonstrated a significant improvement in the mixtures’ mechanical properties; therefore, its application is highly successful and an environmentally friendly solution.

Effect of organic acids on fermentation quality and microbiota of horseshoe residue and corn protein powder

This experiment aimed to investigate the impact of malic acid (MA) and citric acid (CA) on the nutritional composition, fermentation quality, rumen degradation rate, and microbial diversity of a mixture of apple pomace and corn protein powder during ensiling. The experiment used apple pomace and corn protein powder as raw materials, with four groups: control group (CON), malic acid treatment group (MA, 10 g/kg), citric acid treatment group (CA, 10 g/kg), and citric acid + malic acid treatment group (MA, 10 g/kg + CA, 10 g/kg). Each group has 3 replicates, with 2 repetitions in parallel, subjected to mixed ensiling for 60 days. The results indicated: (1) Compared to the CON group, the crude protein content significantly increased in the MA, CA, and MA + CA groups (p < 0.05), with the highest content observed in the MA + CA group. The addition of MA and CA effectively reduced the water-soluble carbohydrate (WSC) content (p < 0.05). Simultaneously, the CA group showed a decreasing trend in NDFom and hemicellulose content (p = 0.08; p = 0.09). (2) Compared to the CON group, the pH significantly decreased in the MA, CA, and MA + CA groups (p < 0.01), and the three treatment groups exhibited a significant increase in lactic acid and acetic acid content (p < 0.01). The quantity of lactic acid bacteria increased significantly (p < 0.01), with the MA + CA group showing a more significant increase than the MA and CA groups (p < 0.05). (3) Compared to the CON group, the in situ dry matter disappearance (ISDMD) significantly increased in the MA, CA, and MA + CA groups (p < 0.05). All three treatment groups showed highly significant differences in in situ crude protein disappearance (ISCPD) compared to the CON group (p < 0.01). (4) Good’s Coverage for all experimental groups was greater than 0.99, meeting the conditions for subsequent sequencing. Compared to the CON group, the Shannon index significantly increased in the CA group (p < 0.01), and the Simpson index increased significantly in the MA group (p < 0.05). However, there was no significant difference in the Chao index among the three treatment groups and the CON group (p > 0.05). At the genus level, the abundance of Lentilactobacillus in the MA, CA, and MA + CA groups was significantly higher than in the control group (p < 0.05). PICRUSt prediction results indicated that the metabolic functional microbial groups in the CA and MA treatment groups were significantly higher than in the CON group (p < 0.05), suggesting that the addition of MA or CA could reduce the loss of nutritional components such as protein and carbohydrates in mixed ensilage. In conclusion, the addition of malic acid and citric acid to a mixture of apple pomace and corn protein powder during ensiling reduces nutritional losses, improves fermentation quality and rumen degradation rate, enhances the diversity of the microbial community in ensiled feed, and improves microbial structure. The combined addition of malic acid and citric acid demonstrates a superior effect.

Effects of Plant Growth Regulators on Micropropagation of Black Pepper (Piper nigrum) in Commercial-Scales

Aim: Black pepper (Piper nigrum) is renowned as a popular spice used to flavor dishes around the world and is also an agricultural export commodity that brings high economic value to many countries. However, traditional propagation methods by seed, cuttings, layering, and grafting reveal many limitations such as the low multiplication coefficient, the low survival rate of seedlings, and especially when there is an outbreak of disease, the ability to control the disease source is exceedingly difficult, leading to the spread of disease. Therefore, an alternative method through apical meristem micropropagation has been applied to contribute to sustainable and disease free pepper cultivars. Propagation by apical shoot meristem gives high propagation efficiency, good and uniform seedling quality, and can be propagated in large numbers in a brief period. Methodology: This study is conducted to contribute to optimizing the process of micropropagation of pepper plants to meet the needs of producing excellent quality seedlings, serving the Vietnamese pepper industry. The two steps in the micropropagation sequence studied in this project are shoot formation and bud cluster multiplication. In this study, we investigate the effects of BA (6-Benzylaminopurine) and NAA (α-Naphthalene acetic acid) in shoot formation. Regenerated shoots were then cultured in MS media supplemented with different combinations of Kinetin (6-Furfurylaminopurine) and NAA in bud cluster multiplication experiment. Results: The results showed that the apical shoot meristem cultured on MS medium (Murashige and Skoog, 1962) supplemented with 0.5 mg L-1 NAA and 2.0 mg L-1 BA was the optimal medium for shoot formation rate (100%). Besides, MS culture medium supplemented with Kinetin 1.5 mg L-1 + NAA 0.5 mg L-1 produced superior results in bud cluster multiplication compared with other treatments (8, 20 buds) after 8 weeks (about 2 months). Conclusion: BA, Kinetin and NAA give important rules on micropropagation of black pepper in commercial-scale.

Characterization of a Food‐Safe Colorimetric Indicator Based on Black Rice Anthocyanin/PET Films for Visual Analysis of Fish Spoilage

ABSTRACTThe safety of food products is of prime importance for consumers and manufacturers. Many means can be used to validate a food product's safety before it is consumed. This study is about the preparation, characterization and evaluation of a generally recognized as safe (GRAS) sensitive colorimetric sensor that detects volatile gases (TVB‐N) resulting from fish spoilage, thus indicating the pH variation of packaged fish products. This is performed by coating a thin layer of ink sensors on the surface of the supporting matrix (corona‐treated PET). Various visual pH indicators were prepared based on black rice anthocyanin as an FDA‐approved dye. Black rice, which contains more than 80% cyanidin‐3‐glucoside, is the most prevalent anthocyanin component. Because of its low toxicity and high concentration, it can be utilized as a natural food colourant. pH indicators based on black rice can show distinct colours in various pH: from red (low pH) to violet (4–6) and deep purple/blue (6–7), blue (7–9) to yellowish/light brown (9–13) throughout the acid–base reaction by the analyte. The ink formulation was prepared by incorporating a binder system (PVOH‐PEG) for higher surface wettability, a crosslinking agent (citric acid) for higher adhesion, an antifoaming agent (natural vanillin) and acetic acid as a pH fixing agent. Corona treatments affected substrate surface chemistry in this study. The samples with thermal treatment passed the ASTM D3330 tape test, the 8000 passages for dry sponge and the 25 passages for wet sponge through the abrasion method. Anthocyanin concentration in formulated ink based on calculation by UV–vis spectra is 0.240 mg/100 g. Sensitivity tests towards TVB‐N gases were carried out at a temperature of 4°C to evaluate the performance of colorimetric films with formulated ink along with thermal treatment (temperature: 165°C, time: 5 min) using the volatile gases emitted by the fish sample inside the package.

Mechanical and Water Barrier Properties of Inhomogeneous Clay Nano-Particles Reinforced Thermoplastic Starch

This research investigated the development of biodegradable bioplastic as a possible replacement for petroleum-based plastics, which constitute a serious environmental hazard. These hazards include but are not limited to flooding resulting from blocked sewage and danger to aquatic life in marine environments. The solution casting method was used to blend inhomogeneous kaolinite clay nano-particles with distilled water, starch, dilute acetic and nitric acids to produce different compositions of thermoplastic starch (TPS)/Clay composites with clay reinforcements ranging from 2.5 to 10 wt.%. The composites were characterized using an X-ray diffraction (XRD), and the mechanical and water absorption properties were determined. The result revealed a 9-fold improvement in the tensile strength (0.72 MPa), flexural strength increased 5-fold (3.34 MPa), and hardness increased 2-fold (23.56 HVN) as well as a reduction in water absorption by 3-fold (6.63%) when compared to the control. Furthermore, the 10 wt.% clay content composite showed the highest mechanical properties. The significant improvement in the listed properties was attributed to a reduction in crystallinity and the formation of new chemical bonds between the thermoplastic starch and the nano-clay. It was observed that the properties of the composites can be further enhanced if a synchronized machine blender (such as an extruder) is employed.

Enhancing probiotic impact: engineering Saccharomyces boulardii for optimal acetic acid production and gastric passage tolerance.

Saccharomyces boulardii has been a subject of growing interest due to its potential as a probiotic microorganism with applications in gastrointestinal health, but the molecular cause for its probiotic potency has remained elusive. The recent discovery that S. boulardii contains unique mutations causing high acetic acid accumulation and inhibition of bacterial growth provides a possible clue. The natural S. boulardii isolates Sb.P and Sb.A are homozygous for the recessive mutation whi2S270* and accumulate unusually high amounts of acetic acid, which strongly inhibit bacterial growth. However, the homozygous whi2S270* mutation also leads to acetic acid sensitivity and acid sensitivity in general. In the present study, we have constructed a new S. boulardii strain, derived from the widely therapeutically used CMCN I-745 strain (isolated from the pharmaceutical product Enterol), producing even higher levels of acetic acid while keeping the same tolerance toward low pH as the parent Enterol (ENT) strain. This newly engineered strain, named ENT3, has a homozygous deletion of ACH1 and strong overexpression of ALD4. It is also able to accumulate much higher acetic acid concentrations when growing on low glucose levels, in contrast to the ENT wild-type and Sb.P strains. Moreover, we show the antimicrobial capacity of ENT3 against gut pathogens in vitro and observed that higher acetic acid production might correlate with better persistence in the gut in healthy mice. These findings underscore the possible role of the unique acetic acid production and its potential for improvement of the probiotic action of S. boulardii.IMPORTANCESuperior variants of the probiotic yeast Saccharomyces boulardii produce high levels of acetic acid, which inhibit the growth of bacterial pathogens. However, these strains also show increased acid sensitivity, which can compromise the viability of the cells during their passage through the stomach. In this work, we have developed by genetic engineering a variant of Saccharomyces boulardii that produces even higher levels of acetic acid and does not show enhanced acid sensitivity. We also show that the S. boulardii yeasts with higher acetic acid production persist longer in the gut, in agreement with a previous work indicating competition between probiotic yeast and bacteria for residence in the gut.