Search Results (32)

Heparin, the naturally occurring, highly sulfated glycosaminoglycan drug, acts as a regulator of diverse biological processes by interacting with diverse specific proteins through a variety of sulfated structural motifs with varying degrees of affinity. Despite its rarity, 3-O-sulfation of glucosamine plays a key role in several biological activities, especially anticoagulant activity mediated by interaction with antithrombin (AT), a mechanism that has been extensively studied. The present work primarily focuses on sequences containing 3-O-sulfated glucosamine that are not involved in anticoagulant activity, analysing their relative abundance and structural environments in heparins derived from distinct animal origins. Three heparin samples derived from bovine, ovine and porcine intestinal mucosa (BMH, OMH and PMH) were fractionated by affinity chromatography on AT-Sepharose into no affinity (NA) and high affinity (HA) fractions. Parent heparins and derived NA and HA fractions were enzymatically depolymerised using either a cocktail of heparinases I, II and III or heparinase II alone, and the resulting mixtures of di- and oligosaccharides were analysed by liquid chromatography coupled with mass spectrometry. Digestion with heparinase II, which preserves heparin sequences containing 3-O-sulfated glucosamine, produced a series of 3-O-sulfated trisaccharides containing two glucosamine residues either side of a uronic acid, located at the non-reducing end (NRE) of heparin chains. Treatment with a heparinase cocktail, which cleaves these trisaccharides, released NRE glucosamine monosaccharides, including 3-O-sulfated species. Notably, both the number of NRE trisaccharide species and the overall proportion of 3-O-sulfated monosaccharides were markedly higher in OMH compared to BMH or PMH. Using ¹H/¹³C bi-dimensional nuclear magnetic resonance spectroscopy, the higher proportion of NRE 3-O-sulfated glucosamine in ovine heparin relative to bovine and porcine heparins was found to be preferentially located in chains lacking affinity for AT. These results offer new insights into the localisation of most 3-O-sulfated heparin sequences that are not associated with anticoagulant activity and, importantly, also reveal a possible structural marker that indicates the ovine origin of heparin. Additionally, a tetrasaccharide species was detected that may be indicative of a highly sulfated AT-binding site in BMH. Full article

Veterinary metabolomics has redefined bile acids (BAs) from simple digestive surfactants to systemic endocrine signals within a microbial–host metabolic axis. This review aims to evaluate how BA dysmetabolism acts as a central pathogenic factor in canine and feline disease. We analyze the BA pool’s integrity, which depends on a specialized functional guild, primarily Peptacetobacter hiranonis, responsible for 7α-dehydroxylation. We delineate two principal pathological profiles: (1) microbial collapse, characterized by secondary bile acid (SBA) depletion and compromised farnesoid X receptor (FXR) and Takeda G protein-coupled receptor 5 (TGR5) signaling, which exacerbates inflammation in chronic enteropathy (CE), protein-losing enteropathy (PLE), and exocrine pancreatic insufficiency (EPI); and (2) hepato-biliary spillover, wherein host-induced dysfunction results in primary bile acid (PBA) excess. Recent data have linked these disruptions to skeletal health, feline renal fibrosis, cardiac remodeling in myxomatous mitral valve disease (MMVD), and neuroinflammation in epilepsy and hepatic encephalopathy. The discovery of microbially conjugated bile acids (MCBAs) and microbial extracellular vesicles (MEVs) reveals highly specific, vesicle-mediated communication pathways impacting systemic health. Diagnostic protocols should prioritize functional profiling, including the dysbiosis index (DI), serum conjugated BA analysis, and SBA/PBA ratios. Clinical management is moving beyond empirical fecal microbiota transplantation (FMT), towards precision synthetic microbial consortia (SynComs), neuroprotective BAs like tauroursodeoxycholic acid (TUDCA), and molecular postbiotics to restore the collaborative metabolome. Full article

The study comprises an in-depth characterization of compositional groups of the liquid by-products obtained from the pyrolysis of swine manure at 500 °C, with the aim of providing an alternative and efficient approach for the valorisation of this waste stream, alongside with the production of biogas and char, the latter of which can be further converted into activated carbon. Two samples were considered: de-watered cake and solid product from anaerobic digestion of swine manure. Biocrude oils were fractionated into weak acidic, strong acidic, alkaline and neutral oil fractions. Subsequently, the neutral oil fraction was separated into paraffinic–naphthenic, slightly polar and polar fractions. All fractions were analyzed by GC–MS. The major identified compositional groups were: (i) for de-watered cake: steroids (40.7%), fatty acids, FAs (23.7%) and n-alkenes/n-alkanes (23.3%); (ii) for solid product from anaerobic digestion: FAs (31.0%), phenols/methoxy phenols (26.6%), n-alkenes/n-alkanes (10.8%) and steroids (10.6%). A variety of short-chain FAs (i.e., linear saturated, mono- and di-unsaturated, cis (i-), trans (ai-), isoprenoid, phenyl alkanoic, amongst others) and methyl esters (FAMEs) were identified as well. FA distribution, nC₁₂–nC₂₀, was similar for both manures studied with nC₁₆ and nC₁₈ as major compounds. FAMEs (nC₁₄–nC₂₈, with even carbon number dominance) in the slightly polar fraction of both samples were accompanied by considerable amounts of oleic (nC_18:1) and linoleic (nC_18:2) acids, and corresponding methyl esters. Hydrocarbons, i.e., n-alkenes/n-alkanes, were in the range of nC₁₅–nC₃₄, with nC₁₈ maximizing. Anaerobically digested manure has resulted in (i) an increase in the portion of longer homologues of hydrocarbons and FAMEs and (ii) the appearance of new FAs series of long chain members nC_22:1–nC_26:1, ω-9. The comprehensive analysis of the biocrude oils obtained from the slow pyrolysis of swine manure indicates their potential for use as biodiesel additives or as feedstock to produce value-added materials. Full article

β-alanine has been shown to significantly improve nitrogen utilization efficiency in beef cattle, but its impact on growth performance remains unclear. This study involved 36 healthy 18-month-old Simmental crossbred bulls with similar weights (627 ± 41 kg). The cattle were divided into two groups, with each group comprising six replicates of three animals. While the control group received the basal diet, the treatment group was administered an additional 96 g/d/cattle rumen-protected β-alanine (RP-β-Ala). The study was conducted over a 35-day period, which included an initial 7 days for adaptation. At the end of the trial, body weight was recorded, and samples were collected. Results show that RP-β-Ala enhanced average daily gain (p = 0.065) and crude protein (CP) digestibility (p = 0.065) and reduced gain-to-feed ratio (p = 0.078). Analysis of rumen microbiota revealed that RP-β-Ala positively modulated the rumen microbiota by enriching beneficial genera such as Prevotella, Treponema, and Selenomonas. This enrichment increased volatile fatty acid production and nitrogen utilization efficiency, as evidenced by elevated ruminal ammonia-N and microbial CP levels, along with decreased serum urea nitrogen. Metabolomics identified key alterations in arachidonic acid metabolism, specifically the upregulation of metabolites 14,15-DiHETrE and prostaglandin D2, and enhanced antioxidative capability indicated by increased serum total antioxidant capacity (T-AOC). Concurrently, RP-β-Ala reduced serum TNF-α levels. This reduction was achieved by suppressing harmful bacteria like Thermoactinomyces and Saccharopolyspora, along with inhibiting their polyamine synthesis, specifically spermine and spermidine. Collectively, these effects alleviated oxidative stress and inflammation. These findings demonstrate that RP-β-Ala enhances beef cattle growth through improved energy supply and antioxidant capacity. Full article

The replacement of fishmeal with plant-based proteins in aquafeeds is key for sustainable aquaculture but may compromise filet pigmentation in rainbow trout (Oncorhynchus mykiss), a quality trait dependent on astaxanthin (Ax) deposition and metabolism. This study aimed to assess the effects of graded fishmeal replacement with a plant protein blend on Ax retention, pigmentation, lipid metabolism, and gut microbiota composition. Rainbow trout were fed three isoenergetic diets containing 60%, 36%, or 12% fishmeal, each supplemented with equal amounts of natural Ax from Haematococcus pluvialis, for 12 weeks. Ax retention, pigmentation, plasma metabolites, lipid digestibility, and distal intestinal microbiota were evaluated. The high plant protein diet (12% fishmeal) significantly reduced Ax concentrations in filet and plasma and decreased dorsal and belly pigmentation scores (p < 0.05). It also lowered plasma cholesterol and bile acid levels by 18–30%, reduced di-esterified Ax digestibility by 15%, and lipid absorption efficiency by 12%. The gut microbiota shifted significantly, with a marked reduction in Bacillaceae, positively correlated with Ax retention and pigmentation. Fish fed High Plant Meal diets exhibited impaired performance parameters, along with reduced lipid accumulation in the liver. High plant meal compromises Ax bioavailability by altering cholesterol–bile acid metabolism and gut microbiota, impairing Ax absorption. However, moderate inclusion of plant meal preserved pigmentation, underscoring the need for dietary cholesterol management and microbiota modulation in plant-based aquafeeds. Full article

Irritable Bowel Syndrome (IBS) is a functional disorder of the digestive system. Its global prevalence varies widely, estimated at up to 10%, due to differences in diagnostic criteria, cultural factors, and dietary patterns. Dietary interventions have emerged as first-line strategies for symptom management, with the low-FODMAP (fermentable oligo-, di-, and monosaccharide and polyol) diet demonstrating robust efficacy in reducing gastrointestinal symptoms by minimizing poorly absorbed, fermentable carbohydrates. However, concerns regarding the long-term nutritional adequacy, potential alterations in gut microbiota composition, and sustainability of the low-FODMAP diet have prompted the need for more integrative and nutritionally balanced dietary models. In contrast, the Mediterranean Diet (MD), rich in fruits, vegetables, legumes, whole grains, nuts, olive oil, and omega-3 fatty acids from fish, is widely recognized for its anti-inflammatory, cardiometabolic, and potential anticarcinogenic effects. Notably, adherence to the MD has been associated with favorable gut microbial profiles and reduced risk of colorectal and other gastrointestinal cancers. However, the high FODMAP content of many MD components limits its immediate compatibility with IBS dietary protocols. This review explores the evidence supporting the integration of MD principles into a low-FODMAP framework for the dietary management of IBS. Rather than proposing a new model, we synthesize existing literature, including recent clinical findings, and offer practical, evidence-informed guidance for tailoring a Mediterranean-style low-FODMAP diet that supports both symptom management and long-term nutritional health. Integrating MD principles into the low-FODMAP approach offers a promising strategy to enhance the nutritional quality, sustainability, and long-term efficacy of dietary management in IBS. Full article

This on-farm study explored the effects of diverse pasture systems and seaweed bio-stimulants (AgriSea NZ Seaweed Products, Paeroa, New Zealand) on sheep performance, metabolic health, milk composition, and carcass characteristics. A 3 × 2 factorial design was used to compare three pasture systems; ryegrass-white clover (RW), a 23-species diverse mix (DI), and functionally diverse strip swards (ST), with (SW) or without (CO) a seaweed bio-stimulant. Ninety pregnant ewes were stratified by live weight and allocated across six treatment groups (15 ewes per treatment). Lambing occurred on treatment paddocks. At weaning, 90 lambs (15 per treatment) were selected based on body weight and sex balance to continue through to finishing. Pasture chemical composition differed among treatments: ST had lower fibre (neutral detergent fibre, NDF; acid detergent fibre, ADF) than RW and DI, while SW increased dry matter digestibility (DMD) and metabolisable energy (ME), and reduced NDF and ADF (p < 0.05). Strip pastures improved lamb average daily gain (ADG) by 17% from lambing to weaning compared to DI, and by 14% from weaning to finishing compared to RW (p < 0.05). Seaweed bio-stimulant treatment enhanced lamb ADG by up to 12% and improved carcass traits, including loin and shoulder yields (p < 0.05). Ewes and lambs on seaweed-treated pastures exhibited lower serum non-esterified fatty acid (NEFA) concentrations (p < 0.05), indicating better energy balance. Milk from ST and/or SW treated ewes had elevated omega-6 fatty acids and essential amino acids, suggesting enhanced nutritional value. These findings demonstrate that combining botanical diversity with natural bio-stimulants can improve animal growth, metabolic health, and product quality, offering a promising strategy for sustainable and welfare-oriented sheep production systems. Full article

The increasing interest in fermented foods stems from their health benefits, mediated by foodborne microorganisms. This study aimed to characterize the fermentative microbiota of Pecorino di Picinisco, a traditional Italian cheese made from ovine raw milk, and to evaluate the probiotic and technological potential of selected lactic acid bacteria strains. Three strains representative of the different species found (Lactococcus lactis, Lactiplantibacillus plantarum and Latilactobacillus curvatus) were chosen and analyzed. All three strains were able to adhere to human intestinal Caco-2 cells, were resistant to simulated in vitro digestion and significantly prolonged the lifespan of Caenorhabditis elegans, used as a simplified in vivo model, with respect to the commercial probiotic strain Lacticaseibacillus rhamnosus GG. The L. plantarum Pic37.4 strain was particularly promising; therefore, its cell-free supernatant was employed to evaluate the antimicrobial activity against indicator strains of foodborne and intestinal pathogens or spoilage bacteria. The results demonstrated the effectiveness of the supernatant against all strains tested, with the strongest effect on the intestinal pathogen enterotoxigenic Escherichia coli K88. In addition, the inhibitory effect on pathogen adhesion to intestinal mucosa was investigated on Caco-2 cells, resulting in a significant reduction in adhesion mediated by the L. plantarum Pic37.4 supernatant. The antimicrobial properties of the L. plantarum strain were confirmed in vivo in C. elegans. These promising results lay the ground for further investigations aimed at substantiating the probiotic and technological potential of the L. plantarum Pic37.4 investigated in this work. Full article

Carob syrup is a brown, thick syrup produced from carob pulp that can be directly consumed or used as a sweetener, which also finds applications in folk medicinal practices. In this work, the quali–quantitative phenolic profile of five different carob syrups was elucidated before and after in vitro gastro–intestinal digestion. Moreover, the anti-oxidant properties of undigested and digested carob syrups were investigated. A total of 75 phenolic compounds were identified in undigested carob syrups. The most important phenolic compound in all the samples was gallic acid, the concentration of which ranged between 54.28 and 117.73 mg/100 g. Additional compounds belonging to the classes of hydroxybenzoic acids (in particular glycosylated gallic acid derivatives), hydroxycinnamic acids, and flavonoids (especially flavonols) were also identified. During in vitro gastric digestion, gallic acid mono- and di-hexosides were diglycosylated, releasing gallic acid, which was further degraded in ellagic acid through oxidative polymerization in the intestinal phase of the digestion. Ellagic acid was the major compound detected after in vitro gastro–intestinal digestion of carob syrups. With few exceptions, the anti-oxidant properties of carob syrup were preserved even after digestion. Carob syrup can be considered an important source of phenolic compounds with demonstrated positive effects on human health. Full article

A predigested product from arachidonic acid oil (ARA) and docosahexaenoic acid (DHA) oil in a 2:1 (w/w) ratio has been developed and evaluated in an in vitro digestion model. To produce this predigested lipid mixture, first, the two oils were enzymatically hydrolyzed up to 90% of free fatty acids (FFAs) were achieved. Then, these two fatty acid (FA) mixtures were mixed in a 2:1 ARA-to-DHA ratio (w/w) and enzymatically esterified with glycerol to produce a mixture of FFAs, mono-, di-, and triacylglycerides. Different glycerol ratios and temperatures were evaluated. The best results were attained at 10 °C and a glycerol-to-FA molar ratio of 3:1. The bio-accessibility of this predigested mixture was studied in an in vitro digestion model. A total of 90% of the digestion product was found in the micellar phase, which contained 30% monoacylglycerides, more than 50% FFAs, and a very small amount of triacylglycerols (3% w/w). All these data indicate an excellent bio-accessibility of this predigested mixture. Full article

Guayusa tea is derived from the leaves of the Ilex guayusa Loes. plant, which is native to the Amazon rainforest. Beyond its pleasant sensory properties, Guayusa tea is rich in antioxidants, phenolics, and minerals. In this study, the effects of infusion time, temperature, and solvent conditions on the color, antioxidant capacity, total phenolic content, phenolic profile, and antimicrobial activity of Guayusa (Ilex guayusa Loes.) tea were investigated. Guayusa tea samples were prepared using two different solvents, ethanol and water, with 4, 6, and 8-h infusions at 60 and 70 °C. Liquid chromatography, diode array detection, and electrospray ionization tandem mass spectrometry (LC-DAD-ESI-MS/MS) were used to determine a comprehensive profile of phenolic compounds and to detect differences due to infusion conditions. Moreover, after the Guayusa tea infusion with the highest bioactive properties was determined, the effects of in vitro gastrointestinal digestion on the total phenolic content, antioxidant capacity, and phenolic compounds of the Guayusa tea infusion were measured. Phenolic profile analysis identified 29 compounds, among which chlorogenic acid and its derivatives were predominant. The increase in infusion time was correlated with an elevation in total phenolic content. Significant differences were observed between water and ethanol infusions of Guayusa in terms of phenolics and antioxidants. The total amount of phenolic compounds in the samples prepared with both solvents was found to increase after oral intake, depending on the digestion stage; meanwhile, the amounts of flavonoid compounds and di-O-caffeoylquinic acid derivatives decreased during digestion. Full article

With rapid industrialization in Gazipur areas of Bangladesh, untreated industrial effluents have been polluting rice soils which could exert potential ecological risk. Therefore, four different types of industries including chemical (SL), textile and paints (MIX), dyeing (CK), and sweater and dyeing (RD) were selected to monitor the intensity of heavy metal pollution in rice soils and ecological risk assessment. The di-acid digestion method was used for the determination of Pd, Cd, and Ni, and the DTPA extraction method was used for Fe, Zn, and Cu. ArcGIS was used to visualize the spatial patterns of heavy metal pollution, and different pollution indices were calculated to assess the ecological risk. The highest concentration (mg kg⁻¹) of Cd (0.72), Pb (104.20), and Ni (5.02) was found in soils of the MIX industrial area. The highest concentration (mg kg⁻¹) of Fe (147.65) and Zn (11.27) was found in the SL industry, while the highest Cu (7.67) was found in the CK industry. It was evident from the spatial distribution that the soils of paddy fields adjacent to the different industries are more contaminated than background soil. Although the potential ecological risk of heavy metal was low, different pollution indices indicated low to high pollution. Thus, the adjacent rice field soil of different industries is being contaminated by different heavy metals which may raise ecological risk. Full article

White mugwort (Artemisia lactiflora Wall.), a traditional Chinese medicine, has been widely consumed in different forms for health care purposes. In this study, the in vitro digestion model of INFOGEST was used to investigate the bioaccessibility, stability, and antioxidant activity of polyphenols from two different forms of white mugwort, including dried powder (P 50, 100, and 150 mg/mL) and fresh extract (FE 5, 15, and 30 mg/mL). During digestion, the bioaccessibility of TPC and antioxidant activity were influenced by the form and ingested concentration of white mugwort. The highest bioaccessibility of the total phenolic content (TPC) and relative antioxidant activity were found at the lowest P and FE concentrations, as calculated relative to the TPC and antioxidant activity of P-MetOH and FE-MetOH based on the dry weight of the sample. Post-digestion, in comparison to P, FE had higher bioaccessibility (FE = 287.7% and P = 130.7%), relative DPPH radical scavenging activity (FE = 104.2% and P = 47.3%), and relative FRAP (FE = 673.5% and P = 66.5%). Nine compounds, 3-caffeoylquinic acid, 5-caffeoylquinic acid, 3,5-di-caffeoylquinic acid, sinapolymalate, isovitexin, kaempferol, morin, rutin, and quercetin, identified in both samples were modified during digestion, yet still provided strong antioxidant activity. These findings suggest that white mugwort extract possesses a higher polyphenol bioaccessibility, showing great potential as a functional ingredient. Full article

Plectranthus ornatus Codd, the genus Plectranthus of the Lamiaceae family, has been used as traditional medicine in Africa, India and Australia. Pharmacological studies show the use of this plant to treat digestive problems. In turn, leaves were used for their antibiotic properties in some regions of Brazil to treat skin infections. The present study examines the anti-inflammatory, antioxidant and cytotoxic effects of the halimane and labdane diterpenes (11R*,13E)-11-acetoxyhalima-5,13-dien-15-oic acid (HAL) and 1α,6β-diacetoxy-8α,13R*-epoxy-14-labden-11-one (PLEC) and the forskolin-like 1:1 mixture of 1,6-di-O-acetylforskolin and 1,6-di-O-acetyl-9-deoxyforskolin (MRC) isolated from P. ornatus on lung (A549) and leukemia (CCRF-CEM) cancer cell lines, and on normal human retinal pigment epithelial (ARPE-19) cell line in vitro. Additionally, molecular docking and computational approaches were used. ADMET properties were analysed through SwissADME and proTox-II—Prediction. The results indicate that all tested compounds significantly reduced the viability of the cancer cells and demonstrated no cytotoxic effects against the non-neoplastic cell line. The apoptosis indicators showed increased ROS levels for both the tested A549 and CCRF-CEM cancer cell lines after treatment. Furthermore, computational studies found HAL to exhibit moderate antioxidant activity. In addition, selected compounds changed mitochondrial membrane potential (MMP), and increased DNA damage and mitochondrial copy number for the CCRF-CEM cancer cell line; they also demonstrated anti-inflammatory effects on the ARPE-19 normal cell line upon lipopolysaccharide (LPS) treatment, which was associated with the modulation of IL-6, IL-8, TNF-α and GM-CSF genes expression. Docking studies gave indication about the lowest binding energy for 1,6-di-O-acetylforskolin docked into IL-6, TNF-α and GM-CSF, and 1,6-di-O-acetyl-9-deoxyforskolin docked into IL-8. The ADMET studies showed drug-likeness properties for the studied compounds. Thus, halimane and labdane diterpenes isolated from P. ornatus appear to offer biological potential; however, further research is necessary to understand their interactions and beneficial properties. Full article

Several studies have shown the ability of yeast to consume peptides as a nitrogen source in single-peptide containing media. However, a suitable and cost-effective methodology to study the utilization of peptides by yeast and other microorganisms in a complex peptide mixture has yet to be put forward. This article addresses this issue by presenting a screening methodology for tracking the consumption of peptides by yeast during alcoholic fermentation. As a peptide source, the methodology makes use of an in-house prepared peptide-mapped bovine serum albumin (BSA) proteolytic digest, which was applied to a synthetic grape must. The peptide uptake was analyzed using high-throughput ultra-high-pressure liquid chromatography coupled to data-independent acquisition-based ion mobility separation-enabled high-resolution mass spectrometry (UPLC-DIA-IMS-HRMS) analysis. The relative changes of abundance of 123 di- to hexapeptides were monitored and reported during fermentations with three commercial wine strains, demonstrating different uptake kinetics for individual peptides. Using the same peptide-mapped BSA hydrolysate, the applicability of an untargeted workflow was additionally assessed for peptide profiling in unelucidated matrixes. The comparison of the results from peptide mapping and untargeted analysis experiments highlighted the ability of untargeted analysis to consistently identify small molecular weight peptides on the length and amino acid composition. The proposed method, in combination with other analytical techniques, such as gene or protein expression analysis, can be a useful tool for different metabolic studies related to the consumption of complex nitrogen sources by yeast or other microorganisms. Full article