Growth performance,digestibility, and plasma metabolomic profiles of Saanen goats exposed to different doses of aflatoxin B1

Aflatoxin contamination of feed poses a great risk to the global dairy industry. Analyzing the aflatoxin B₁ (AFB₁)-induced metabonomic changes in ruminants and screening potential biomarkers for early diagnosis of AFB₁ exposure is urgently needed. Here, the effects of different doses (0, 50, and 500 μg/kg of the diet, dry matter basis) of AFB₁ exposure on digestibility and performance of Saanen goats were studied, and a comprehensive untargeted metabolomic analysis was performed to reveal plasma metabonomic changes caused by the AFB₁ exposure. In the current study, AFB₁ exposure decreased total-tract nutrient digestibilities, nitrogen retention, total weight gain, and average daily gain of Saanen goats in a dose-dependent manner. Untargeted metabolomics revealed alterations in the plasma metabolome. A total of 3,310 and 1,462 ion peaks were obtained in positive and negative ion modes, respectively. Based on the screening criteria, 1,338 differential metabolites were detected between control and low-dose AFB₁ (50 μg/kg) groups, 1,358 metabolites differed between control and high-dose AFB₁ (500 μg/kg) groups, and 58 metabolites differed among all groups. Pathway analyses showed that choline metabolism in cancer and glycerophospholipid metabolism were significantly affected by the AFB₁ treatments. Moreover, dysregulation of amino acid metabolism was also observed in AFB₁ treated goats. The findings provided novel insights into the toxicity of AFB₁ in ruminants. Exploring the underlying molecular causes of the changes may help the development of rapid diagnostic techniques and effective interventions for AFB₁ intoxication.     

Changes in serum metabolites in response to ingested colostrum and milk in neonatal calves,measured by nuclear magnetic resonance-based metabolomics analysis

Uptake of colostrum is of central importance for establishing a passive immunity transfer in neonatal calves. Studies of absorption and transmission of colostral immunoglobulins have been widely reported; however, changes in the serum in response to the absorption of colostral components in neonatal calves have not been completely characterized. Here, a nuclear magnetic resonance-based metabolomics approach was used to investigate the changes in metabolites in ingested colostrum, milk, and serum after neonatal calves were fed colostrum or milk. Twenty-seven neonatal male Holstein calves were assigned to 1 of the following groups: (1) calves not fed colostrum or milk and slaughtered approximately 2 h after birth (control group, n = 6), (2) calves fed colostrum at 1 to 2 h after birth and slaughtered 8 h after birth (n = 6), (3) calves fed 2 colostrum meals (at 1–2 and 10–12 h after birth) and slaughtered 24 h after birth (n = 6), (4) calves fed 3 colostrum meals (at 1–2, 10–12, and 22–24 h after birth) and slaughtered 36 h after birth (n = 6), or (5) calves fed 2 milk meals (1–2 and 10–12 h after birth) and slaughtered 24 h after birth (n = 3). Concentrations of valine, leucine, lactate, lysine, and isoleucine were higher and concentrations of lactose were lower in the groups fed colostrum and milk compared with groups not fed colostrum and milk, respectively. Metabolite changes between groups fed or not fed colostrum and milk were similar and may reflect the primary metabolic requirements of ingestion by the small intestine of neonatal calves. Concentrations of serum metabolites choline, valine, leucine, and glutamate were higher in the serum of calves that received colostrum compared with control calves. Furthermore, concentrations of serum phenylalanine, valine, and glutamate were significantly higher, whereas serum concentrations of citrate and very low density lipoproteins were lower in calves that received colostrum compared with calves fed milk. Our results indicate that concentrations of leucine, valine, and glutamate, which were higher in the calves that ingested colostrum, may transfer into the bloodstream, and that these metabolites are associated with health benefits in the neonatal calves that received colostrum. These findings provide novel information to help us understand the mechanism by which colostrum components are metabolized and absorbed in the small intestine and then transferred into bloodstream of neonatal calves.     

Discrepancies among healthy,subclinical mastitic,and clinical mastitic cows in fecal microbiome and metabolome and serum metabolome

Mastitis is generally considered a local inflammatory disease caused by the invasion of exogenous pathogens and resulting in the dysbiosis of microbiota and metabolites in milk. However, the entero-mammary pathway theory may establish a possible link between some endogenous gut bacteria and the occurrence and development of mastitis. In the current study, we attempted to investigate differences in the gut microbiota profile and metabolite composition in gut and serum from healthy cows and those with subclinical mastitis and clinical mastitis. Compared with those of healthy cows, the microbial community diversities in the feces of cows with subclinical mastitis (SM) and clinical mastitis (CM) were lower. Lower abundance of Bifidobacterium, Romboutsia, Lachnospiraceae_NK3A20_group, Coprococcus, Prevotellaceae_UCG-003, Ruminococcus, and Alistipes, and higher abundance of the phylum Proteobacteria and the genera Escherichia-Shigella and Streptococcus were observed in CM cows. Klebsiella and Paeniclostridium were significantly enriched in the feces of SM cows. Several similarities were observed in feces and serum metabolites in mastitic cows. Higher levels of proinflammatory lipid products (20-trihydroxy-leukotriene-B4, 13,14-dihydro-15-keto-PGE₂, and 9,10-dihydroxylinoleic acids) and lower levels of metabolites involved in secondary bile acids (deoxycholic acid, 12-ketolithocholic acid), energy (citric acid and 3-hydroxyisovalerylcarnitine), and purine metabolism (uric acid and inosine) were identified in both SM and CM cows. In addition, elevated concentrations of IL-1β, IL-6, tumor necrosis factor-α and decreased concentrations of glutathione peroxidase and superoxide dismutase were detected in the serum of SM and CM cows. Higher serum concentrations of triglyceride and total cholesterol and lower concentrations of high-density lipoproteins in mastitic cows might be related to changes in the gut microbiota and metabolites. These findings suggested a significant difference in the profile of feces microbiota and metabolites in cows with different udder health status, which might increase our understanding of bovine mastitis.     

Metabolomic profile of milk fermented with Streptococcus thermophilus cocultured with Bifidobacterium animalis ssp. lactis,Lactiplantibacillus plantarum,or both during storage

In this study, the microbial interactions among cocultures of Streptococcus thermophilus (St) with potential probiotics of Bifidobacterium animalis ssp. lactis (Ba) and Lactiplantibacillus plantarum (Lp) in fermented milk were investigated during a storage period of 21 d at 4°C, in terms of acidifying activity (pH and titratable acidity), viable counts, and metabolites. A nontargeted metabolomics approach based on ultra-high-performance liquid chromatography coupled with mass spectrometry was employed for mapping the global metabolite profiles of fermented milk. Probiotic strains cocultured with St accelerated milk acidification, and improved the microbial viability compared with the single culture of St. The St–Ba/Lp treatment manifested a higher bacteria viability and acidification ability in comparison with the St–Ba or the St–Lp treatment. Relative quantitation of 179 significant metabolites was identified, including nucleosides, AA, short peptides, organic acids, lipid derivatives, carbohydrates, carbonyl compounds, and compounds related to energy metabolism. The principal component analysis indicated that St treatment and coculture treatments displayed a complete distinction in metabolite profiles, and Lp had a larger effect than Ba on metabolic profiles of fermented milk produced by cofermentation with St during storage. The heat map in combination with hierarchical cluster analysis showed that the abundance of metabolites significantly varied with the starter cultures over the storage, and high abundance of metabolites was observed in either St or coculture samples. The St-Ba/Lp treatment showed relatively high abundance for the vast majority of metabolites. These findings suggest that the profile of the metabolites characterizing fermented milk samples may depend on the starter cultures, and incorporation of probiotics may considerably influence the metabolomic activities of fermented milks.     

Varying the ratio of Lys:Met while maintaining the ratios of Thr:Phe,Lys:Thr,Lys:His,and Lys:Val alters mammary cellular metabolites,mammalian target of rapamycin signaling,and gene transcription

Amino acids are not only precursors for but also signaling molecules regulating protein synthesis. Regulation of protein synthesis via AA occurs at least in part by alterations in the phosphorylation status of mammalian target of rapamycin (mTOR) pathway proteins. Although the ideal profile of Lys:Met to promote milk protein synthesis during established lactation in dairy cows has been proposed to be 3:1, aside from being the most-limiting AA for milk protein synthesis, the role of Met in other key biologic pathways such as methylation is not well characterized in the bovine. The objective of this study was to determine the influence of increasing supplemental Met, based on the ideal 3:1 ratio of Lys to Met, on intracellular metabolism related to protein synthesis and mTOR pathway phosphorylation status. MAC-T cells, an immortalized bovine mammary epithelial cell line, were incubated (n = 5 replicates/treatment) for 12 h with 3 incremental doses of Met while holding Lys concentration constant to achieve the following: Lys:Met 2.9:1 (ideal AA ratio; IPAA), Lys:Met 2.5:1 (LM2.5), and Lys:Met 2.0:1 (LM2.0). The ratios of Thr:Phe (1.05:1), Lys:Thr (1.8:1), Lys:His (2.38:1), and Lys:Val (1.23:1) were the same across the 3 treatments. Applying gas chromatography–mass spectrometry metabolomics revealed distinct clusters of differentially concentrated metabolites in response to Lys:Met. Lower Phe, branched-chain AA, and putrescine concentrations were observed with LM2.5 compared with IPAA. Apart from greater intracellular Met concentrations, further elevations in Met level (LM2.0) led to greater intracellular concentrations of nonessential AA (Pro, Glu, Gln, and Gly) compared with IPAA and greater essential AA (EAA; Met, Ile, and Leu) and nonessential AA (Pro, Gly, Ala, Gln, and Glu) compared with LM2.5. However, compared with IPAA, mRNA expression of β-casein and AA transporters (SLC7A5, SLC36A1, SLC38A2, SLC38A9, and SLC43A1) and mTOR phosphorylation were lower in response to LM2.5 and LM2.0. Overall, the results of this study provide evidence that increasing Met while Lys and the ratios of Phe, Thr, His, and Val relative to Lys were held constant could increase the concentration and utilization of intracellular EAA, in particular branched-chain AA, potentially through improving the activity of AA transporters partly controlled by mTOR signaling. Because EAA likely are metabolized by other tissues upon absorption, a question for future in vivo studies is whether formulating diets for optimal ratios of EAA in the metabolizable protein is sufficient to provide the desired levels of these AA to the mammary cells.     

Metabolomic analysis of significant changes in Lactobacillus casei Zhang during culturing to generation 4,000 under conditions of glucose restriction

Lactic acid bacteria are being consumed more frequently as awareness of their health benefits has increased. The industrial production of lactic acid bacteria requires a comprehensive understanding of their survival stress, especially regarding changes in metabolic substances in a glucose-limited environment. In the present study, a metabolomic approach was applied to investigate Lactobacillus casei Zhang using cultures from a common ancestor that were permitted to evolve under conditions with normal or glucose-restricted media for up to 4,000 generations. Metabolomic analyses of intracellular and extracellular differential metabolites under De Man, Rogosa and Sharpe broth (2% vol/vol glucose; Oxoid Ltd., Basingstoke, UK) and glucose-restricted (0.02% vol/vol glucose in De Man, Rogosa and Sharpe broth) conditions were performed at generations 0, 2,000, and 4,000 and revealed 23 different metabolites. Myristic acid, ergothioneine, Lys-Thr, and palmitamide contents exhibited significant reductions between 0 and 4,000 generations, whereas nicotinate, histidine, palmitic acid, l-lysine, urocanate, thymine, and other substances increased. The dynamics of the pathways involved in AA metabolism, including glycine, serine, and threonine metabolism, histidine metabolism, lysine degradation, and arginine and proline metabolism, were also a focus of the present study. There were also changes in several other metabolic pathways, including vitamin B₆, thiamine, nicotinate, and nicotinamide, according to generation time. Additionally, in the present study we screened for key metabolites involved in the glucose-restricted response and provided a theoretical basis for comprehensively revealing the regulatory mechanisms associated with L. casei Zhang glucose restriction at the metabolic level. These findings also provide novel ideas and methods for analyzing the glucose-restricted stress response at the metabolic level.     

Superchilled,chilled and frozen storage of Atlantic mackerel (Scomber scombrus) – effect on lipids and low molecular weight metabolites

Quality changes in Atlantic mackerel fillets during superchilled, chilled and frozen storage focusing on lipid quality, colour and changes in low molecular weight metabolites relevant for quality and safety were studied. Low formation of oxidation products was observed in chilled (up to 7 days), superchilled (up to 14 days) and frozen storage (up to one year). Only slight formation of TBARS was measured in the samples which correlated with the increasing fillet yellowness. The profile of low molecular weight compounds, analysed by high-resolution NMR, showed clear grouping of the different samples according to both treatment and storage time. The increase in K-value was highest in the chilled samples, reaching a K-value of 93 ± 3% on day 5, exceeding the proposed limit of human consumption (80%) while superchilling reached a K-value of ca 90 % after 14 days. Frozen samples showed acceptable quality in the whole storage period.     

Examination of Serum Metabolome Altered by Dietary Carbohydrate,Milk Protein,and Soy Protein Interventions Identified Novel Metabolites Associated with Blood Pressure: The ProBP Trial

Scope

This study aims to discover metabolites of dietary carbohydrate, soy and milk protein supplements and evaluate their roles in blood pressure (BP) regulation in the protein and blood pressure (ProBP), a cross-over trial.

Methods and results

Plasma metabolites are profiled at pre-trial baseline and after 8 weeks of supplementation with carbohydrate, soy protein, and milk protein, respectively, among 80 ProBP participants. After Bonferroni correction (α = 6.49 × 10⁻⁴), dietary interventions significantly changed 40 metabolites. Changes of erucate (22:1n9), an omega-9 fatty acid, are positively associated with systolic BP changes (Beta = 1.90, p = 6·27 × 10⁻⁴). This metabolite is also associated with higher odds of hypertension among 1261 participants of an independent cohort (odds ratio per unit increase = 1.34; 95% confidence interval: 1.07–1.68). High levels of acylcholines dihomo-linolenoyl-choline (p = 4.71E-04) and oleoylcholine (p = 3.48E-04) at baseline predicted larger BP lowering effects of soy protein. Increasing cheese intake during the trial, as reflected by isobutyrylglycine and isovalerylglycine, reduces the BP lowering effect of soy protein.

Conclusions

The study identifies molecular signatures of dietary interventions. Erucate (22:1n9) increases systolic BP. Acylcholine enhances and cheese intake reduces the BP lowering effect of soy protein supplement.     

基于代谢组学技术的玛咖产地鉴别

采用代谢组学的方法,基于超高效液相色谱-四极杆串联飞行时间质谱对不同产地的玛咖进行产地鉴别和各产地代谢物差异分析。通过正交偏最小二乘判别分析成功区分玛咖的5 个产地,并对各个产地玛咖的代谢物进行靶标和非靶标代谢组学分析,其中靶标代谢组学分析共鉴定出28 种代谢物,包括玛咖酰胺、玛咖烯、芥子油苷、生物碱、甾醇、多酚和黄酮,非靶标代谢组学分析鉴定出玛咖的产地标志物。结果表明代谢组学方法可以用于玛咖产地鉴别以及全组分的鉴定。     

基于代谢组学研究荡秋千运动下普洱茶茶褐素对大鼠代谢综合征改善作用

分析比较荡秋千运动对普洱茶茶褐素（theabrownin,TB）在高脂高糖高盐饮食大鼠血清内的代谢组学变化。采用1H-核磁共振（1H-nuclear magnetic resonance,1H-NMR）代谢组学技术对14 组大鼠的血清进行分析,归属、指认图谱中化合物,进行正交偏最小二乘-判别分析（orthogonal partial least squares-discrimination analysis,OPLS-DA）,发现其中的差异及规律。结果显示：常规代谢综合征（metabolic syndrome,MS）模型组大鼠血清中异亮氨酸、亮氨酸、缬氨酸、3-羟基丁酸相对含量显著低于常规正常对照组（P＜0.05）,丙氨酸、N-乙酰基-糖蛋白、谷氨酸、二甲基甘氨酸、甜菜碱、牛磺酸、精氨酸、糖原、甘油相对含量则显著高于常规正常对照组（P＜0.05）。荡秋千TB高剂量组大鼠的3-羟基丁酸相对含量显著高于荡秋千MS模型组,而低密度脂蛋白/极低密度脂蛋白（low density lipoprotein/very low density lipoprotein,LDL/VLDL）、乳酸、丙氨酸、精氨酸相对含量则显著低于荡秋千MS模型组。与常规正常对照组相比,荡秋千后的正常对照组大鼠血清中高密度脂蛋白（high density lipoprotein,HDL）、异亮氨酸、3-羟基丁酸相对含量显著提高,而二甲基甘氨酸、精氨酸、糖原、α-葡萄糖相对含量则显著降低（P＜0.05）。与MS模型组相比,荡秋千MS模型组大鼠血清中LDL/VLDL、异亮氨酸相对含量显著降低,而丙氨酸、乳酸相对含量则显著升高（P＜0.05）。研究结果表明,荡秋千可明显降低血清LDL/VLDL相对含量,升高HDL相对含量,促进血糖代谢;荡秋千运动与TB处理具有协同增效的作用,在预防代谢综合征和心脑血管疾病方面具有很大潜力。