Concentrations Of Acids Research Articles

Metabolites and Free Fatty Acids in Japanese Black Beef During Wet Aging

Abstract: Background: Japanese Black beef is known for its high intramuscular fat content, an important factor in its distinctive Wagyu aroma. Wet aging, which involves vacuum-packing meat and storing it at low temperatures, enhances flavor, texture, and tenderness and is essential for maintaining and improving meat quality. In this study, changes in metabolites and lipid profiles were investigated during the wet aging of Japanese Black and Holstein beef. Methods/Results: Gas chromatography–mass spectrometry identified 113 metabolites in Japanese Black beef and 94 in Holstein beef, with significant increases in metabolites like aspartic acid and maleic acid over the aging period. Regarding lipid composition, total free fatty acids significantly increased with wet aging, with Japanese Black beef showing significantly higher concentrations of oleic and linoleic acids than Holstein beef. Additionally, lipid analysis by liquid chromatography–mass spectrometry revealed a reduction in specific phospholipids, particularly lysophosphatidylcholine (LPC) and lysophosphatidylethanolamine (LPE), with notable decreases in LPC (18:1), LPC (18:2), LPE (18:1), and LPE (18:2). Conclusions: These results suggest that wet aging influences the stability of membrane lipids, facilitating the degradation of phospholipids into free fatty acids, and improving the flavor of Japanese Black beef.

Oxalic Acid Supplementation in Different Hemicellulose Diets Affects In Vitro Rumen Fermentation by Regulating Nutritional Digestibility, Microbial Diversity and Metabolic Pathways

Determining hemicellulose (HM) degradation is crucial for evaluating the nutritional value of ruminant diets. Our previous study showed that oxalic acid (OA) regulates rumen fermentation. Building on this research, the present study examined the effects of OA supplementation in different hemicellulose diets on sheep rumen fermentation, microbial diversity, and metabolite production in vitro. Diets with low and high HM levels (10.3% and 17%, respectively) and supplemented with seven concentrations of OA (0, 2.5, 5, 10, 20, 40, 80 mg/kg DM) were evaluated. Tests were conducted under both low (HM10.3%) and high (HM17%) hemicellulose conditions; however, the addition of 10 mg/kg DM oxalic acid could have better effects under low hemicellulose (HM10.3%), with higher concentrations of acetic, propionic, and butyric acids, as well as total acids. A 2 × 2 factorial design was used to collect rumen fluid after 12 h of fermentation to analyze microbial populations and metabolites. OA supplementation at 10 mg/kg DM significantly increased the relative abundances of several bacterial genera, including Prevotella, Butyrivibrio, Ruminococcus, Sharpea, RFN20, Bulleidia, Olsenella, and Bifidobacterium (p < 0.05). A positive correlation was observed between Butyrivibrio and Sharpea and the production of isobutyric and isovaleric acids (p < 0.01), indicating that these bacteria play a role in volatile fatty acid (VFA) production. Furthermore, rumen metabolites involved in mineral absorption and lipid metabolism, including α-tocopherol, L-glutamic acid, and ginkgolide B, were upregulated. In summary, supplementation with oxalic acid in HM diets alters rumen fermentation, enhances nutrient digestibility, promotes microbial diversity, and influences metabolic pathways. Thus, OA supplementation should be tailored to specific dietary conditions for optimal effects.

Antialgal Effects of Nonanoic and Palmitic Acids on Microcystis aeruginosa and the Underlying Mechanisms

Algal blooms caused by Microcystis aeruginosa are a common occurrence and pose significant threats to freshwater ecosystems. This study investigates the antialgal effects and underlying mechanisms of two plant-derived fatty acids, nonanoic acid and palmitic acid, on Microcystis aeruginosa. The results show that the inhibitory effects of both fatty acids on M. aeruginosa increase with higher concentrations. Algal recovery occurs when nonanoic acid concentrations are below 0.5 mg/L and palmitic acid concentrations are below 50 mg/L. Acute toxicity tests indicate that the safe concentrations of nonanoic acid and palmitic acid are below 1.87 mg/L and 263.3 mg/L, respectively. The inhibitory effect of nonanoic acid is more pronounced under conditions of pH 5.5, 15 °C temperature, 0.75 mg/L nitrogen, and 2 mg/L phosphorus, with inhibition efficiency remaining unaffected by increased light intensity. Both fatty acids exert their strongest inhibitory effects in the early stages of addition (0–8 days), causing cell death and the release of extracellular organic matter primarily consisting of aromatic compounds and proteins. Oxidative stress analysis reveals that high concentrations of fatty acids can cause irreversible damage to the algae’s antioxidant defense system. These findings provide valuable insights for the prevention and control of cyanobacterial blooms, which can help promote the sustainable development of freshwater ecosystems.

Taste adaptations in blood-feeding arthropods: mechanisms and ecological implications.

Hematophagous arthropods rely on taste mechanisms to navigate host selection, feeding, mating, and oviposition. These behaviors are driven by environmental taste cues, which shape acceptance or aversion depending on their valence. Positive stimuli, like low concentrations of salts, sugars, amino acids, and nucleotides, promote feeding and oviposition, while negative stimuli, including high salt, bitter compounds, and nociceptive chemicals, trigger avoidance to prevent hazards. Species-specific adaptations enable blood feeders to overcome ecological challenges. Understanding their behavioral, neuronal, and molecular taste mechanisms aids in developing targeted vector control strategies, such as repellents, toxic baits, and oviposition deterrents, to disrupt disease transmission.

Cytokinin B-Mo-based Product Influences the Source-to-sink Dynamics and Nonstructural Carbohydrate Contents in Hydroponic Lettuce Plants

An understanding of how amendments influence the sink-to-source relationship in leafy crops can be used to optimize plant resource allocation for enhanced growth and quality. Variations in growth rates and carbon pools across individual leaves or groups of leaves at similar developmental stages allow us to comprehend plant strategies of carbon allocation and partitioning. We hypothesized that products enhancing the carbon source-to-sink relationship during leaf development can increase growth and dry matter accumulation. This project aimed to determine whether exogenous applications of a cytokinin-B-Mo-based product during the leaf development of lettuce plants impact the carbon source-to-sink relationship, thus influencing plant growth and quality. The experiment was a complete randomized design with two treatments: a negative control and the application of the product twice during the growing cycle. Each experimental unit consisted of a deep-water culture reservoir with three lettuce plants. Destructive sampling was conducted five times throughout the cycle. At each sampling time (n = 4 per experimental run), the phenological stage was determined, and measurements of root and shoot length, root and shoot dry matter, leaf length, leaf width, leaf area, chlorophyll contents, and nonstructural carbon contents were performed. These data were used to estimate growth indices. The results indicated that the cytokinin-B-Mo-based product increased the number of true unfolded leaves by 1 ± 0.4 and the overall size of the lettuce head by 9%. The treated lettuce reached marketable size 4 days earlier than that of the control treatment. Statistically significant differences were observed in shoot and root dry matter accumulation and foliar length and width at some sampling points. Some growth indices indicate an increase in leaf surface area investment and enhanced conversion efficiency of assimilates into biomass in plants treated with the product. Plants exhibiting these alterations had higher sucrose and total soluble sugar contents. There was a noticeable pattern of higher concentrations of nonstructural carbohydrates, proteins, and amino acids in the leaves compared with those in the roots across all plants and treatments. Overall, the cytokinin-B-Mo-based product appears to strengthen the source-to-sink relationship during lettuce development, resulting in a high-quality plant within a shorter timeframe.

Lactobacillus salivarius metabolite succinate enhances chicken intestinal stem cell activities via the SUCNR1-mitochondria axis.

The activity of intestinal stem cells (ISCs) can be modulated by Lactobacillus, which subsequently affects the mucosal absorptive capacity. However, the underlying mechanisms remain unclear. In this study, a total of 189 Hy-Line Brown chickens (Gallus) were randomly assigned to one of seven experimental groups (n = 27 per group). These groups included a control group, a vehicle group (MRS group), a Lactobacillus salivarius group, a L. salivarius supernatant group, and three succinate treatment groups with various dosages. Each group was further subdivided into three replicates, with 9 chickens per replicate. The results indicate that the administration of Lactobacillus salivarius supernatant to laying hens notably increased the mRNA abundance of the amino acid transporters oligopeptide transporter 1 (PepT1) and sodium-dependent neutral amino acid transporter (B0AT). Metabolomic analyses indicated that the supernatant contains a high concentration of organic acids. Among them, succinate could enhance mRNA abundance of PepT1, B0AT and excitatory amino acid transporters 3 (EAAT3) both in vivo and in vitro. Accordingly, succinate could accelerate intestinal epithelial turnover, as indicated by the increased levels of cyclin-dependent kinase 2 (Cdk2) mRNA and proliferating cell nuclear antigen protein (PCNA), as well as ISC differentiation-related protein leucine-rich repeat containing G protein-coupled receptor 5 (LGR5). Furthermore, succinate treatment was shown to elevate the levels of mitochondrial fusion proteins optic atrophy 1 (OPA1) and translocase of outer mitochondrial membrane 20 (TOMM20), resulting in increased local ATP levels. However, pretreatment with NF-56-EJ40, a succinate receptor antagonist, attenuated the effects of succinate on OPA1, TOMM20, and ATP levels, alone with the reducing LGR5 and PCNA levels. Collectively, succinate, a metabolite of L. salivarius, activates the SUCNR1-mitochondria axis in ISCs, facilitating mitochondrial ATP synthesis, promoting ISC activity, and ultimately enhancing mucosal absorptive capacity.

Biomass production and silage quality of ensiled BRS Capiaçu elephant grass at different regrowth ages and residue heights

The aim of this study was to determine the optimal combination of regrowth age and residue height of BRS Capiaçu elephant grass that yields the best balance between the dry mass production (DMP) of the harvested forage and silage quality. Four regrowth ages (75, 90, 105, and 120 days) and two residue heights (10 and 50 cm) were evaluated. The experimental design used a randomized complete block design with a 4 × 2 factorial scheme and 3 replicates. The regrowth age led to a linear increase in DMP. As regrowth age advanced, there were linear increases in the dry matter (DM) concentration, cell wall constituents, and fraction C (acid detergent insoluble nitrogen) of crude protein and reductions in the in vitro digestibility of DM and neutral detergent fiber (IVNDFD) in both the plant and its silage. Lower pH values were observed in silage made from plants harvested at 10 cm. No significant effects of the studied factors were found on the concentrations of organic acids in the silage, except for propionic acid. Management using a 105-day regrowth period and a 10-cm residue height resulted in a better balance between the dry mass production, fermentative profile and nutritional value of silage.

Plasma amino acid profiles of dogs with the hepatocutaneous syndrome and dogs with other chronic liver diseases.

Dogs with hepatocutaneous syndrome (HCS) have marked plasma hypoaminoacidemia, but its occurrence in dogs with chronic liver diseases not associated with HCS (non-HCS CLD) is unknown. To determine if plasma hypoaminoacidemia occurs in dogs with non-HCS CLD, compare plasma amino acid (PAA) profiles between dogs with non-HCS CLD and HCS, and define a sensitive and specific PAA pattern for diagnosing HCS. Data were collected from client-owned dogs, a prospective cohort of 32 with CLD and 1 with HCS, and a retrospective cohort of 7 with HCS. Prospective study. Dogs with chronic serum liver enzyme increases were recruited after hepatic biopsy. Plasma amino acid profiles were measured using high-performance liquid chromatography. Plasma amino acid concentrations were compared between dogs with non-HCS CLD and HCS. Regression analysis was performed to identify a unique PAA pattern for HCS diagnosis. Twelve dogs each with vacuolar hepatopathy or chronic hepatitis and 8 dogs with congenital disorders (primary hypoplasia of the portal vein or ductal plate malformations) were enrolled. Compared to non-HCS CLD dogs, HCS dogs had significantly lower plasma concentrations of several amino acids. Regression analysis revealed that glutamine, glycine, citrulline, arginine, and proline concentrations less than 30% of the mean reference value had 100% sensitivity, specificity for diagnosing HCS. Generalized plasma hypoaminoacidemia does not accompany non-HCS CLD. Concentrations of 5 specific amino acids less than 30% of the mean reference value can serve as a noninvasive biomarker for diagnosing HCS.

Integration of tissue distribution, PK-PD modeling and metabolomics reveals inflammatory-immune response alterations in Gaultheria leucocarpa var. yunnanensis alleviating rheumatoid arthritis.

Gaultheria leucocarpa var. yunnanensis, a distinguished member of the Gaultheria Kalm ex L. in the Ericaceae family, has been traditionally employed in the southwestern regions of China for the efficacious treatment of rheumatoid arthritis (RA). The anti-RA fraction (ARF) derived from Gaultheria leucocarpa var. yunnanensis has been previously demonstrated to effectively alleviate RA in vivo and in vitro. This research endeavor is dedicated to surveying the pharmacokinetic (PK) processes of ARF within plasma and tissues, profiling its metabolites in vivo, discerning the material foundation of its therapeutic efficacy, and delineating its anti-RA mechanisms. The prototype components and metabolites of ARF in plasma and seven tissues of RA rats were analyzed by LC-MSn. Advanced LC-MS/MS and HPLC-DAD methodologies were developed to investigate the plasma PK profiles and tissue distribution characteristics of MSTG-A, MSTG-B, and Gaultherin in both RA model rats and healthy controls. A panel of four cytokines (TNF-α, IL-1, IL-6, and IL-2) was selected as pharmacodynamic (PD) biomarkers and quantified using ELISA. The PK, PD, and PK-PD modeling of ARF were skillfully constructed by combining WinNonlin with Matlab software, enabling a comprehensive analysis of the interrelationships between components and effect markers. A non-targeted plasma metabolomics approach employing LC-QE-MS was utilized to insight into the underlying mechanisms of ARF alleviating RA. The quantity and diversity of identified prototypical components and metabolites of ARF in model rat plasma increased over time. The spleen exhibited the highest number of metabolites and prototypical compounds of ARF. The UPLC-QQQ-MS/MS and HPLC-DAD method were developed and validated for the quantification of three chemical markers in rat plasma and tissues, respectively. Three effective components (MSTG-B, MSTG-A, and Gautherin) demonstrated linear dynamics in plasma and tissues at an oral dosage of 3 g/kg ARF. The PK-PD models involving three components and four inflammatory cytokines aligned with the one company model, demonstrating a linear correlation through compartmental modeling and curve fitting analysis. Significant variations were identified in the concentrations of various amino acids and lipid metabolites among the CON, ARF, and MTX groups in comparison to the MOD group, which are intricately linked to the inflammation-immunity response. The three components displayed favorable bioavailability and were rapidly eliminated in RA rats, collectively exerting an anti-RA effect. The mechanism by which ARF mitigates RA is associated with the modulation of inflammation-immunity related metabolic pathways. The spleen may serve as the target tissue for ARF attenuating RA. These findings provide a robust foundation for rationalizing intervention strategies, elucidating biological mechanisms, and advancing the clinical application of ARF in the amelioration of RA.

Nickel-induced multimetal uptake in two microalgal species (Chlorella sorokiniana and Chlamydomonas reinhardtii) and its effect on growth and lipid unsaturation.

As the concern for Ni contamination in the aquatic environment escalates, efforts for microalgal use in environmental monitoring and bioremediation are increasing. This study aims to evaluate the potential of Chlorella sorokiniana and Chlamydomonas reinhardtii for Ni bioremediation by investigating their physiological stress responses in Ni-contaminated environments. The analysis focuses on how Ni(II) uptake affects cell growth, nutrient metal homeostasis, and lipid unsaturation levels, as these parameters are critical indicators of metabolic stability and resilience essential for effective bioremediation. The microalgae were grown under mixotrophic conditions in a tris-acetate-phosphate (TAP) medium enriched with Ni(II), at concentrations (1-6 mg∙L-1) exceeding those typically found in wastewater, providing insights into metal stress under severe contamination conditions. Even though increased uptake of Ni(II) was observed for both algal species, accompanied by growth suppression at high Ni(II) concentrations, multi-elemental trace analysis revealed a significant, Ni concentration-dependent, uptake of growth media essential metals as well. Specifically, for both algal species, Zn uptake concentrations increased by approximately 20 times when going from control cultures, with no Ni(II) added, to cultures incubated with increasing Ni(II) concentrations. Overall, Zn uptake was determined to be approximately 3 orders of magnitude higher than Ni(II) uptake when high concentrations of Ni(II) were present, making Zn the metal with the most significant uptake. Similar uptake trends were observed for Cu and Co for both algal species, with Cu uptake being approximately 2 orders of magnitude higher, while Co remained below the Ni(II) concentrations at high added Ni(II) concentrations. For Chlorella sorokiniana, increased Fe uptake relative to Ni(II) uptake was observed (2 orders of magnitude higher), as was the case for Mn (1 order of magnitude higher). This induced increase in uptake of some of the growth media metals was attributed to their liberation from ethylenediaminetetraacetic acid (EDTA) in tris-acetate-phosphate (TAP) medium, following the addition of Ni(II), which has a higher stability constant (Kf) with EDTA and was added at concentrations comparable or higher than those of the other metals. Calculated levels of free Ni(II) and free metals in the medium matched the observed metal uptake trends as determined using multielemental inductively coupled plasma mass spectrometry. Negative ion electrospray mass spectrometry also revealed that EDTA-metal complexes in the TAP media decreased as Ni(II) concentrations increased. The lipid unsaturation level and relative ω-3 fatty acids concentration of both microalgal species, based on 1H Nuclear Magnetic Resonance analysis, decreased with increasing Ni(II) concentration, with the decrease being more pronounced at Ni(II) incubation concentrations of 4 and 6 ppm. Unsaturation levels for individual lipid classes [monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), and sulfoquinovosyldiacylglycerol (SQDG)] in Chlamydomonas reinhardtii cells were also studied using positive ion mode electrospray mass spectrometry. At the highest Ni(II) concentrations, an overall reduction in unsaturation levels was observed for all 3 lipid classes, indicating a significant impact of elevated metal ion concentrations on membrane fluidity and therefore on cellular physiology and metabolism. Comparison of the two microalgal species under Ni-enriched conditions shows that Chlorella sorokiniana exhibits greater tolerance to the metal-induced stress under study than Chlamydomonas reinhardtii, suggesting its higher efficiency for the bioremediation in Ni-contaminated environments.

A Review on Fish Oil Extraction from Fish by-Product as Sustainable Practices and Resource Utilization in the Fish Processing Industry

The fish processing industry generates significant by-products, such as viscera, skin, bones, and heads, which are valuable for producing food, medicinal products, energy, and industrial feedstock. Fish oil, rich in omega-3 polyunsaturated fatty acids like eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), is widely used in nutritional supplements and other applications. Among these by-products, fish viscera contain the highest concentration of oil, making them an ideal target for extraction due to their cost-effectiveness and environmental benefits. Extracting oil from fish by-product helps reduce environmental pollution and promotes sustainable practices by fully utilizing fish resources. This holistic approach contributes to waste reduction and resource efficiency in the fish processing industry. By incorporating sustainable principles into extraction processes - such as using environmentally friendly solvents, implementing efficient solvent recovery systems, and ensuring compliance with environmental regulations - companies can enhance the sustainability of their operations while extracting valuable components. As demand for fish-based food products rises, effective extraction of fish oil and fishmeal from by-products becomes increasingly important. Various extraction methods, including physical, chemical, and biological approaches, are essential for separating solids, oil, and water to recover valuable components like EPA and DHA. Optimizing these processes and combining different methods can achieve high concentrations of polyunsaturated fatty acids (PUFAs) in fish oil, ranging from 65% to 80%. Emphasizing maximum PUFA content highlights the potential to enhance the quality and nutritional value of fish oil extracted from by-products while advancing sustainability in the fish processing industry.

Development of an enzymatic method for efficient production of DHA-enriched phospholipids through immobilized phospholipase A1 in AOT-water reverse micelles.

The demand for omega-3 polyunsaturated fatty acids (PUFAs), particularly docosahexaenoic acid (DHA), has been steadily increasing due to their significant health benefits. Traditional methods for producing DHA-enriched phospholipids often suffer from low efficiency and high costs. In this study, we developed an efficient enzymatic process to prepare phospholipid-DHA, which used immobilized phospholipase A1 to catalyze transesterification in AOT-water reverse micelle systems. Initially, high concentrations of free fatty acids were produced via acid hydrolysis of algae oil followed by crystallization. Among six evaluated reverse micelle systems, one was selected for further optimization. The substrate/enzyme ratio, temperature, reaction time, and water content were optimized using single-factor experiments and response surface methodology. To enhance cost-efficiency and eco-friendly practices, substrate recycling was implemented to maximize substrate utilization. This study established a comprehensive process chain for the preparation of phospholipid-DHA, promoting its industrial production and providing a reference for the production of other phospholipid products.

Effect of medium-chain fatty acids supplementation on feed intake, rumen fermentation, blood profile, and milk production of dairy cows in the transition period.

Medium-chain fatty acids (MCFAs) are rapidly utilized by the liver as an energy source. However, the effects of feeding MCFAs to dairy cows during the transition period and the subsequent effect on postpartum performance is not well clear. Therefore, this study was to evaluate the effects of MCFAs supplementation on feed intake, rumen fermentation, blood profile, and milk production in dairy cows during their transition period. Twenty-one multiparity Holstein cows were randomly assigned to 2 dietary groups: one group received a diet supplemented with MCFAs calcium salts (MCFA-Ca) from 3 weeks relative to expected parturition to 5 weeks postpartum (MCFA; n = 10), and the other group received the same diet without supplements (control; n = 11). MCFA-Ca containing 80% caprylic acid and 20% capric acid were added to the mixed diet at 1.5% dry matter. During the close-up period (from 3 weeks relative to expected parturition until parturition), cows were fed a TMR with restricted intake to provide 100% of the metabolic energy requirement. Subsequently, the cows were fed a TMR ad libitum from parturition to 5 weeks postpartum. The dry matter intake and body weight did not differ throughout the experimental period between groups. Rumen fermentation and protozoa counts remained unaffected during the experimental period. Supplementation of MCFA had no effect on the plasma concentrations of nonesterified fatty acids and total ketone bodies in cows throughout the experimental period. On the other hand, postpartum plasma glucagon concentration was higher in the MCFA group than in the control group, and milk yield tended to increase in the MCFA group. In conclusion, our results suggested that MCFA-Ca supplementation enhances the catabolic state by affecting pancreatic hormone secretion, resulting in an increase in milk yield during the fresh period without an excessive negative energy balance in dairy cows.

Breed and trace mineral source influence the performance of beef heifers during periods of nutrient restriction and grazing forage at early vegetative stage.

We evaluated the effects of breed and mineral source on heifer performance during periods of nutrient restriction and grazing. On day -7, ½ Angus × ½ Nelore (ANE) and Nelore (NE) heifers (12 heifers per breed; body weight, BW = 264 ± 35kg; age = 15 ± 1 mo) were assigned to individual drylot pens to receive ad libitum Tifton 85 (Cynodon sp.) hay and white salt for 7 days. On day 0, within each breed, heifers were randomly assigned (2 × 2 factorial arrangement) to receive protein supplementation (0.20% of BW; dry matter, DM) added with sulfate (SUL) or hydroxychloride (HYD) sources of Cu, Mn, and Zn from day 0 to 49. From day 0 to 34, hay DM intake was restricted to 50% of the ad libitum intake recorded from day -7 to -1. On day 35, heifers were transferred to individual pastures to graze Tifton 85 forage at early vegetative stage until day 49. No effects of breed × mineral source × day and breed × mineral source were detected (P ≥ 0.11). Nelore heifers had greater (P ≤ 0.02) average daily gain (ADG) from day 0 to 35 and day 0 to 49 compared to ANE heifers. Cumulative diarrhea incidence, fecal pH and total days of diarrhea symptoms did not differ (P ≥ 0.19) between breeds. Nelore heifers had greater (P ≤ 0.05) serum concentrations of insulin-like growth factor 1 (IGF-1) from day 35 to 45 but had less (P ≤ 0.05) serum concentrations of cortisol and haptoglobin on day 42 and 45, respectively. Serum concentrations of urea N were greater (P ≤ 0.05) for NE vs. ANE heifers on days 0 and 42 and were less (P ≤ 0.05) for NE vs. ANE on days 38, 45 and 49. Heifers supplemented with HYD had (P ≤ 0.05) greater ADG from day 0 to 35, lower cumulative diarrhea incidence and percentage of heifers exhibiting ≥ 2 days of diarrhea from day 36 to 49, less serum concentrations of non-esterified fatty acids (NEFA) on day 35, and less serum concentrations of NEFA, cortisol and urea N on day 38 compared to SUL heifers. In summary, breed influenced serum concentrations of haptoglobin, cortisol, urea N, insulin and IGF-1, and the growth of beef heifers during nutrient restriction, but did not impact growth and incidence of diarrhea during periods of grazing forage at early vegetative stage. Regardless of breed, replacing sulfate with hydroxychloride sources of Cu, Mn, and Zn led to minimal reductions on serum concentrations of NEFA, cortisol and urea N, enhanced growth during nutrient restriction, and reduced diarrhea incidence during grazing of early vegetative forage.

Hypoxia-inducible factor 2α mediates nonesterified fatty acids and hypoxia-induced lipid accumulation in bovine hepatocytes.

Ketosis is a metabolic disorder frequently occurring in the perinatal period, characterized by elevated circulating concentrations of nonesterified fatty acids (NEFA) due to negative energy balance, resulting in fatty liver in dairy cows. However, the mechanism of hepatic steatosis induced by high concentrations of NEFA in ketosis remains unclear. Hypoxia-inducible factor 2α (HIF-2α), which mediates adaptation to hypoxic stress, plays a critical role in regulating lipid metabolism. In this study, we investigate whether HIF-2α is involved in NEFA-driven hepatic lipid accumulation in dairy cows with ketosis. Liver and blood samples were collected from 10 healthy cows (blood β-hydroxybutyrate (BHB) concentration <1.2 mM) and 10 ketotic cows (blood BHB concentration >3.0 mM with clinical symptoms) with similar lactation numbers (median = 3, range = 2 to 4) at 3 to 9 d in milk (median = 6). In cows with ketosis, serum concentrations of NEFA and β-hydroxybutyrate (BHB) were greater but serum concentrations of glucose were lower. Moreover, hepatic triglyceride content increased significantly. In the liver of ketotic cows, which was accompanied by upregulated HIF-2α expression. To determine the potential association among hypoxia, HIF-2α, and the formation of hepatocellular steatosis in vitro, we isolated hepatocytes from healthy calves for the following experiments. First, hepatocytes were treated with 0, 0.6, 1.2, or 2.4 mM NEFA (52.7 mM stock NEFA solution was diluted in RPMI-1640 basic medium supplemented with 2% fatty acid-free BSA to achieve the specified concentrations) for 18 h, showing that HIF-2α expression and cellular hypoxia occurred in a dose-dependent manner. Next, hepatocytes were infected with HIF-2α (encoded by EPAS1) small interfering RNA (Si-HIF-2α) for 48 h and then treated with 1.2 mM NEFA for 18 h. Results indicated that silencing HIF-2α decreased NEFA-induced lipid accumulation in bovine hepatocytes. Subsequently, hepatocytes treated with or without NEFA were placed in an AnaeroPack System, mimicking a hypoxic condition, for 0, 12, 18, or 24 h. Results showed that hypoxia could induce and further exacerbate lipid accumulation in bovine hepatocytes. Meanwhile, normal or NEFA-treated hepatocytes were cocultured with or without PT2385, a specific HIF-2α inhibitor, showing that hypoxia promoted steatosis through HIF-2α. Activating transcription factor 4 (ATF4) is an endoplasmic reticulum (ER) stress and hypoxia-inducible transcription factor. Here, bovine hepatocytes were treated with NEFA or hypoxia following transfecting ATF4 small interfering RNA (Si-ATF4), which demonstrated that ATF4 knockdown alleviated the extent of lipid accumulation in bovine hepatocytes. Besides, we found that ATF4 expression was correlated with HIF-2α levels in both liver tissue and cultured hepatocyte models. Moreover, overexpression of ATF4 weakened the beneficial effects of HIF-2α inhibition. Overall, these data suggest that NEFA-induced hepatic hypoxia significantly contributes to the progression of hepatic steatosis which in turn, intensifies hypoxia and leads to a self-perpetuating cycle of reciprocal causation, further exacerbating hepatic lipid deposition. Additionally, accumulated HIF-2α plays a critical role in this complex-origin steatosis, potentially through ATF4.

СОСТАВ КОМПОНЕНТОВ, ВЛИЯЮЩИХ НА ВКУС ВИНА ИЗ СОРТА КРАСНОСТОП ЗОЛОТОВСКИЙ В ЗАВИСИМОСТИ ОТ ПРОВОДИМЫХ АГРОТЕХНИЧЕСКИХ ПРИЕМОВ

The aim of the study is to identify the features of the composition of biochemical indicators affecting the taste of dry red wines from the autochthonous grape variety Krasnostop Zolotovsky depending on the load of bushes with shoots. Objectives: to conduct a physicochemical and sensory analysis of the experimental wines. Objectives of the study: dry red wines from the technical grape variety Krasnostop Zolotovsky, grown in the vineyards of the All-Russian Research Institute of Viticulture and Winemaking. The studies were conducted at the laboratory for quality control of viticulture and winemaking products of the All-Russian Research Institute for Viticulture and Winemaking, a branch of the Federal State Budgetary Scientific Institution FRANTS. As part of the study, wines were prepared using the microwinemaking method in accordance with the following experimental options: B-1 – with a load of 20 shoots per bush (s/c); B-2 – 25 shoots per bush; B-3 – 30 shoots per bush; B-4 – 35 shoots per bush; B-5 – with a load of 40 shoots per bush. Analysis of the obtained data on the study of the composition of biochemical components affecting the taste of dry red wines from the autochthonous grape variety Krasnostop Zolotovsky depending on the load of the bushes with shoots showed that the sum of organic acids in the studied samples was within 6.7–8.1 g/dm3 with their highest content in Krasnostop Zolotovsky B-4 (35 p/k). The amount of tartaric acid was about 50 % of the total amount of acids in the wine. Variants with a bush load of 30 (B-3) and 35 (B-4) shoots per bush were distinguished by the accumulation of phenolic and coloring substances. The highest content of malic acid was noted in the experimental sample B-5 (load 40 p/k). The ratio of mass concentrations of tartaric and malic acids in all experimental variants was &gt; 2, which contributes to obtaining a more harmonious taste. The highest content of lactic acid was observed in B-3 (1.01 g/dm3) and B-4 (0.85 g/dm3), these same variants were distinguished by the softest and roundest taste. The highest content of reduced and residual extract was noted in B-3 and B-4, and the least amount was in B-2. As a result of sensory analysis, it was revealed that the most complete, meaningful taste and complex aroma were distinguished by the experimental samples Krasnostop Zolotovsky B-3 and B-4 (8.9 points each). In these same variants, the highest content of phenolic, coloring and extractive substances was observed.

GRAZING AND ITS RELATIONSHIP WITH NITROGEN COMPOUNS IN Prosopis laevigata IN NORTHERN MEXICO FORESTS

Mesquite forests Prosopis laevigata functions as a nourishment source mainly for beef cattle. This study was carried out in order to find if mesquite forests are affected by the grazing, particularly on nitrogen components due to its importance in the vegetal growth and tree development. The objective of this study was to determine if the extensive grazing has influence over the concentration of the nitrogen components amino acids and proteins in mesquite forest. Root and stem samples in adult trees were taken monthly during twelve months in two pure mesquite stands, one of them with grazing activity. The total soluble proteins (TSP) and total amino acids (TA) concentrations were determined by visible UV spectrophotometry. The results shown that grazing has assimilation effects in nitrogen components in trees of the mesquite wild populations. The root and stem where there is no grazing activities showed greater concentrations of nitrogen components in comparison with the site where grazing is done as an anthropogenic activity.

Isorhamnetin Ameliorates Non-Esterified Fatty Acid-Induced Apoptosis, Lipid Accumulation, and Oxidative Stress in Bovine Endometrial Epithelial Cells via Inhibiting the MAPK Signaling Pathway

High concentrations of non-esterified fatty acids (NEFAs) in the blood contribute to various metabolic disorders and are linked to endometritis in dairy cows. Isorhamnetin (ISO), a flavonoid found in many plants, is known for its antioxidant, anti-inflammatory, and anti-obesity properties. This study systematically assessed NEFA-induced damage in bovine endometrial epithelial cells (bEECs) and investigated whether ISO alleviates NEFA-induced cell damage and its underlying molecular mechanisms. Our observations revealed that excessive NEFA inhibited proliferation and induced apoptosis in bEECs, accompanied by an increase in the expression of BAX and cleaved caspase-3. We further observed that NEFA could induce lipid accumulation, reactive oxygen species (ROS) generation, and the release of pro-inflammatory factors IL-1β, IL-6, and TNF-α in bEECs. RNA sequencing and Western blot analysis revealed that NEFA induced damage in bEECs by activating MAPK signaling pathway. Notably, ISO treatment ameliorated these effects induced by NEFA, as evidenced by decreased protein levels of BAX, cleaved caspase-3, and PPAR-γ, along with reductions in triglyceride content, ROS generation, and levels of IL-1β, IL-6, and TNF-α. Mechanistically, our experimental results demonstrated that ISO inhibited NEFA-induced activation of MAPK signaling. Overall, ISO shows promise for therapeutic development to address NEFA-related endometritis in dairy cows.

Effects of dietary Acremonium terricola culture on production performance, serum biochemical parameters, egg quality and yolk amino acid contents of Beijing You-chicken

The effects of Acremonium terricola culture (ATC) on production performance, serum biochemical parameters, egg quality and amino acid contents in the yolk of eggs of Beijing You-chicken were conducted in the current study. A total of 216 Beijing You-chickens (330 days old) were randomly divided into 2 groups. The control group (CON) was fed a corn-soybean-based diet, and the experimental group was fed a basal diet supplemented with 0.20% ATC. The pretest period was 7 d, and the experiment period was 8 weeks. The production performance, serum biochemical parameters, egg quality, and the concentrations of amino acids in the yolk of eggs were measured at the 4 weeks (FW, the first stage) and the 8 weeks (EW, the second stage) of the experiment, respectively. Compared with the CON group, there were no significant differences (P > 0.05) in the production performance of the experimental group at the end of four- and eight-week periods of study. The concentration of serum LH, FSH and E2 increased significantly for the ATC group, at both time periods when compared to CON group, while the triglyceride (TG) content was only increased significantly (P < 0.05) in the first stage. The average egg weight, albumen height, and Haugh unit representing egg quality of Beijing You-chickens in the experimental group were increased significantly (P < 0.05) compared with the CON group at both time periods, while the egg shape index and yolk weight were only increased significantly (P < 0.05) in the second stage. The protein content in the yolk was increased significantly at both time periods (P < 0.05). Levels of lecithin and Vitamin A in yolks of the ATC supplemented group increased significantly (P < 0.05) compared to the CON group, at both FW and EW, respectively. The contents of aspartic acid, threonine, methionine, leucine, and arginine were increased significantly in the first stage. In addition, the contents of threonine, glutamine, and valine were increased significantly in the second stage (P < 0.05). Our results suggest that dietary supplementation with 0.20% ATC improves serum biochemical parameters and egg quality in Beijing You-chickens. Future studies should focus on optimizing ATC dosage and exploring its underlying mechanisms for enhanced poultry production.

Daily feeding frequency impacts muscle characteristics and fat deposition in finishing pigs associated with alterations in microbiota composition and bile acid profile

IntroductionFeeding frequency has been shown to affect growth and body composition of the host associated with gut microbiota. It remains unknown whether adjusting feeding frequency could effectively regulate both skeletal muscle development and whole-body lipid metabolism and thus affect carcass composition and feed conversion efficiency. Therefore, this study aimed to explore the effects of feeding frequency on muscle growth, fat deposition, cecal microbiota composition, and bile acid composition in finishing pigs.MethodsSixteen Sichuan-Tibetan black pigs, with an initial weight of 121.50 ± 1.60 kg, were divided into two groups and fed either two meals (M2) or four meals (M4) per day. The trial lasted 30 days. The muscle fiber characteristics, lipid metabolism in adipose tissue, and cecal microbiota and bile acid composition were determined.ResultsThe present study revealed that pigs fed four meals exhibited a lower feed-to-gain ratio, abdominal fat weight, and average backfat thickness (p &amp;lt; 0.05), as well as a higher loin eye area (p = 0.09) and myofiber diameter in the longissimus muscle than their counterparts. The mRNA expression of slow-twitch fiber and myogenesis-associated genes in the longissimus muscle was upregulated, while lipid metabolism-related genes in the backfat were downregulated in the M4 group compared to the M2 group (p &amp;lt; 0.05). The M4 pigs exhibited higher abundances of Firmicutes, Actinobacteriota, Bacillus, Clostridium_sensu_1, and Romboutsia, and lower abundances of Spirochaetota, Verrucomicrobiota, Treponema, and Muribaculaceae in the cecal content than the M2 pigs (p &amp;lt; 0.05). A higher feeding frequency increased the levels of primary bile acids and decreased the concentrations of taurine-conjugated bile acids in the cecal content of pigs (p &amp;lt; 0.05).ConclusionOur research suggested that the M4 feeding pattern, compared to the M2 pattern, promoted muscle growth and reduced fat deposition by enhancing fast- to slow-twitch fiber conversion and myogenesis in the muscle and repressing lipid metabolism in adipose tissue, associated with altered microbiota composition and bile acid profiles.