Objective: In the present study, an liquid chromatography/mass spectrometry (LC/MS) metabolomics approach was performed to investigate potential biomarkers of milk production in high- and low-milk-yield dairy cows and to establish correlations among rumen fluid metabolites. Methods: Sixteen lactating dairy cows with similar parity and days in milk were divided into high-yield (HY) and low-yield (LY) groups based on milk yield. On day 21, rumen fluid metabolites were quantified applying LC/MS. Results: The principal component analysis and orthogonal correction partial least squares discriminant analysis showed significantly separated clusters of the ruminal metabolite profiles of HY and LY groups. Compared with HY group, a total of 24 ruminal metabolites were significantly greater in LY group, such as 3-hydroxyanthranilic acid, carboxylic acids, carboxylic acid derivatives (L-isoleucine, L-valine, L-tyrosine, etc.), diazines (uracil, thymine, cytosine), and palmitic acid, while the concentrations of 30 metabolites were dramatically decreased in LY group compared to HY group, included gentisic acid, caprylic acid, and myristic acid. The metabolite enrichment analysis indicated that protein digestion and absorption, ABC transporters and unsaturated fatty acid biosynthesis were significantly different between the two groups. Correlation analysis between the ruminal microbiome and metabolites revealed that certain typical metabolites were exceedingly associated with definite ruminal bacteria; Firmicutes, Actinobacteria, and Synergistetes phyla were highly correlated with most metabolites. Conclusion: These findings revealed that the ruminal metabolite profiles were significantly different between HY and LY groups, and these results may provide novel insights to evaluate biomarkers for a better feed digestion and may reveal the potential mechanism underlying the difference in milk yield in dairy cows.
BACKGROUND: This study was carried out to assess a biochemical methane potential of giant miscanthus (Miscanthus sacchariflorus) which was a promising candidate energy crop due to a high biomass productivity, in order to utilize as a feedstock for the biogas production. METHODSANDRESULTS: Giant miscanthus was sampled the elapsing drying time of 6 months after harvesting. TS (Total Solid) and VS (Volatile Solid) contents were 94.7 and 90.8%. And CP (Crude Protein), EE (Ether Extracts), and CF (Crude Fiber) contents of giant miscanthus were 1.4, 0.46, and 46.12%, respectively. In the organic composition of giant miscanthus, the NDF (Neutral Detergent Fiber) representing cellulose, lignin, and hemicellulose contents showed 86.88%, and the ADF (Acid Detergent Fiber) representing cellulose and lignin contents was 62.91%. Elemental composition of giant miscanthus showed 47.75%, 6.44%, 41.00%, and 0.28% for C, H, O, and N, respectively, and then, theoretical methane potential was obtained to $0.502Nm^3kg^{-1}-VS_{added}$. Biochemical methane potential was assessed as the range of $0.154{\sim}0.241Nm^3kg^{-1}-VS_{added}$ resulting the lower organic biodegradability of 30.7~48.0%. CONCLUSION: Therefore the development of pretreatment technology of the giant miscanthus was needed for the improvement of anaerobic digestability.
Kim, Mi-Nyeu;Kim, Young Il;Cho, Chunghee;Mayo, Kelly E.;Cho, Byung-Nam
Molecules and Cells
/
v.38
no.12
/
pp.1079-1085
/
2015
Originally, activins were identified as stimulators of FSH release in reproduction. Other activities, including secondary axis formation in development, have since been revealed. Here, we investigated the influence of activin ${\beta}_A$ on the body, including the gastro-intestinal (GI) tract. Initially, the activin ${\beta}_A$ protein was detected in the serum proportional to the amount of pCMV-rAct plasmid injected. The induced level of activin ${\beta}_A$ in muscle was higher in female than male mice. Subsequent results revealed that stomach and intestine were severely damaged in pCMV-rAct-injected mice. At the cellular level, loss of parietal cells was observed, resulting in increased pH within the stomach. This phenomenon was more severe in male than female mice. Consistent with damage of the stomach and intestine, activin ${\beta}_A$ often led to necrosis in the tip of the tail or foot, and loss of body weight was observed in pCMV-rAct-injected male but not female mice. Finally, in pCMV-rAct-injected mice, circulating activin ${\beta}_A$ led to death at supraphysiological doses, and this was dependent on the strain of mice used. Taken together, these results indicate that activin ${\beta}_A$ has an important role outside of reproduction and development, specifically in digestion. These data also indicate that activin ${\beta}_A$ must be controlled within a narrow range because of latent lethal activity. In addition, our approach can be used effectively for functional analysis of secreted proteins.
An enzymatic flour milling process for barley into three major fractions (barley flour, bran-crease-germ and water solubles) was studied. Carbohydrate and protein of barley endosperm could be efficiently solubilized by the digestion process of partially pearled barley with enzymes. Bran, crease and germ were removed from hydrolyzate by filtering through 30-40mesh sieves. And then filtered product was separated into fractions by sedimentation or centrifugation. The most effective digestion of the barley was obtained by the enzyme with higher activities of glucanase and protease under such conditions as barley-water ratio, 1:1.5(W/V) and temperature at $45^{\circ}C$. Total flour yield recovered was approximately 73-76% of the barley, and the portions recovered as bran-crease-germ and water solubles were about 3.6 and 15.8%, respectively.
Jongkeon Kim;Kwanho Park;Sang Yun Ji;Beob Gyun Kim
Journal of Animal Science and Technology
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v.65
no.5
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pp.1002-1013
/
2023
The objectives of the present study were to determine the nutrient digestibility of fish meal, defatted black soldier fly larvae (BSFL), and adult flies and to develop equations for estimating in vitro nutrient digestibility of BSFL for pigs. In vitro digestion procedures were employed to mimic the digestion and absorption of nutrients in the pig intestine. Correlation coefficients between chemical composition and in vitro nutrient digestibility of BSFL were calculated. In Exp. 1, in vitro ileal digestibility (IVID) of dry matter (DM) and crude protein (CP) and in vitro total tract digestibility (IVTTD) of DM and organic matter in defatted BSFL meal were less (p < 0.05) than those in fish meal but were greater (p < 0.05) than those in adult flies. In Exp. 2, CP concentrations in BSFL were negatively correlated with ether extract (r = -0.91) concentration but positively correlated with acid detergent fiber (ADF; r = 0.98) and chitin (r = 0.95) concentrations. ADF and chitin concentrations in BSFL were negatively correlated with IVID of DM (r = -0.98 and -0.88) and IVTTD of DM (r = -1.00 and -0.94) and organic matter (r = -0.99 and -0.98). Prediction equations for in vitro nutrient digestibility of BSFL were developed: IVID of CP (%) = -0.95 × ADF (% DM) + 95 (r2 = 0.75 and p = 0.058) and IVTTD of DM (%) = -2.09 × ADF + 113 (r2 = 0.99 and p < 0.001). The present in vitro experiments suggest that defatted BSFL meal was less digestible than fish meal but was more digestible than adult flies, and nutrient digestibility of BSFL can be predicted using ADF as an independent variable.
Chanjula, P.;Wanapat, M.;Wachirapakorn, C.;Rowlinson, P.
Asian-Australasian Journal of Animal Sciences
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v.17
no.10
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pp.1400-1410
/
2004
Eight crossbred (75% Holstein Friesian) cows in mid-lactation were randomly assigned to a switchback design with a 2x2 factorial arrangement to evaluate two nonstructural carbohydrate (NSC) sources (corn meal and cassava chips) with different rumen degradability and used at two levels of NSC (55 vs. 75%) with protein source (supplied by urea in the concentrate mix). The treatments were 1) Low degradable low level of corn (55%) 2) Low degradable high level of corn (75%) 3) High degradable low level of cassava (55%) and 4) High degradable high level of cassava (75%). The cows were offered the treatment concentrate at a ratio to milk yield at 1:2. Urea-treated rice straw was offered ad libitum as the roughage and supplement with 1 kg/hd/d cassava hay. The results revealed that total DM intake, BW and digestion coefficients of DM were not affected by either level or source of energy. Rumen fermentation parameters; NH3-N, blood urea nitrogen and milk urea nitrogen were unaffected by source of energy, but were dramatically increased by level of NSC. Rumen microorganism populations were not affected (p>0.05) by source of energy, but fungal zoospores were greater for cassava-based concentrate than corn-based concentrate. Milk production and milk composition were not affected significantly by diets containing either source or level of NSC, however concentrate than corn-based concentrate averaging (4.4 and 4.2, respectively). Likewise, income over feed, as estimated from 3.5% FCM, was higher on cassava-based concentrate than corn-based concentrate averaging (54.0 and 51.4 US$/mo, respectively). These results indicate that feeding diets containing either cassava-based diets and/or a higher of oncentrates up to 75% of DM with NPN (supplied by urea up to 4.5% of DM) can be used in dairy rations without altering rumen ecology or animal performance compared with corn-based concentrate.
Microbial growth efficiency in the rumen was studied in sheep given hourly, 31.25 g oaten chaff with either 0.31 and 0.88 g urea or 1.88 and 5.63 g casein (exp. 1) and 33.33 g oaten chaff with 1.04 casein or 0.3, 0.6 and 0.9 g urea or the mixture of the casein and urea (exp. 2). Concentrations of ruminal fluid ammonia increased with increasing nitrogenous supplements. Organic matter digestibility in sacco in the rumen was not different irrespective of N sources. Isoacids and valeric acid increased with increasing ingested casein but decreased with increasing urea intake. Peptide and amino acid pools in ruminal fluid increased with increasing ammonia concentrations (exp. 2) suggesting that proteolytic activity and transportation of peptides and amino acids across microbial membrane of rumen microbes may be regulated by the metabolite mechanism (intracellular amino acids and $NH_4{^+}$, respectively). Densities of total viable and cellulolytic bacteria in ruminal fluid increased with increasing ammonia levels but that of small Entodinia decreased. The density of fungal sporangia growth on oat leaf blades decreased with increasing ammonia concentrations but appeared to remain constant in the presence of casein. Efficiency of net microbial cell synthesis was 15-28% higher when ammonia concentrations increased from 100 to above 200 mg N/l regardless of N sources. In conclusion, supplementation of preformed protein had no effect on rumen digestion and microbial growth efficiency. This could not be accounted for its effect on ruminal fluid ammonia. Increased microbial growth efficiency with increasing ammonia levels may be due to a reduction in the turnover of microbial cells within the rumen.
Cromakalim (BRL 34915), known as an airway smooth muscle relaxant, inhibited the releases of mediators in the antigen-induced mast cell activation. It has been suggested that cromakalim, in part, inhibited mediator releases by inhibiting the initial increase of 1,2-diacylglycerol (DAG) produced by the activation of the other phospholipase system which is different from phosphatidylcholine-phospholipase D pathway. The aim of this study is to further examine the inhibitory mechanism of cromakalim on the mediator release in the mast cell activation. Guinea pig lung mast cells were purified by using enzyme digestion and percoll density gradient. In purified mast cells prelabeled with $[^3H]PIP_2$, phospholipase C (PLC) activity was assessed by the production of $[^3H]$insitol phosphates. Protein kinase C (PKC) activity was assessed by measuring the protein phosphorylated from mast cells prelabeled with $[{\gamma}-32P]ATP$, and Phospholipase $A_2\;(PLA_2)$ activity by measuring the lyso-phosphatidylcholine produced from mast cell prelabeled with 1-palmitoyl-2-arachidonyl $phosphatidyl-[^{14}C]choline$. Histamine was assayed by fluorometric analyzer, and leukotrienes by radioimmunoassay. The PLC activity was increased by activation of the passively sensitized mast cells. This increased PLC activity was decreased by cromakalim pretreatment. The PKC activity increased by the activation of the passively sensitized mast cells was decreased by calphostin C, staurosporine and cromakalim, respectively. The $PLA_2$ activity was increased in the activated mast cells. The pretreatment of cromakalim did not significantly decrease $PLA_2$ activity. These data show that cromakalim inhibits histamine release by continuously inhibiting signal transduction processes which is mediated via PLC pathway during mast cell activation, but that cromakalim does not affect $PLA_2$ activity related to leukotriene release.
The mash of Takjoo, Korean flour wine, is fermented through two brewing processes ; the primary brewing process to saccharify and the main one to produce ethyl alcohol. The activities of acid proteinase (pH3), weak acid proteinase (pH 6), and alkaline proteinase (Ph 80 on the processes are determined with time by the Folin phenol method as a strength of casein digestion. Hydrogen ion concentration, the content of total organic acids, protein, free amino acids and oligopeptides, which effect the activities of proteinase, are also measured. The results are briefly summarized as follows : 1. In general, the activities of acid proteinase and weak acid proteinase in the mesh of primary brewing process are stronger than those in main brewing process. 2. The activities of acid proteinase are remarkably stronger than those of weak acid proteinase in both processes. It reveals that they decrease slowly through the fermentation. Activities of alkaline proteinase are weaker than others. 3. As the raw materials are mixtured, the total amount of organic acids is equivalent to 0.150 mg/ml acetic acid in the mesh of primary brewing process and 0.02 mg/ml acetic acid in the main one. They increase gradually with time. 4. Hydrogen ion concnetration shows 3.9 in the mesh of main brewing process and 3.28 in the primary one. They increase to the maximum in 60-72 hrs., and decrease since 108 hrs. 5. The content of crude protein shows 66.90mg/ml in the mesh of main brewing process, while shows 64.29mg/ml in the mesh of primary one. they decrease slowly with time. it seems that a small content of crude protein, as a substrate, converts into amino acids and soluble nitrogen compounds by proteinase. 6. The content of free amino acids and oligopeptides shows 0.36 mg/ml in the mesh of primary brewing process and 0.24mg/ml in the main brewing process. It is evident that the reason they increase continuously through the fermentation is the effect of proteinase. 7. According to the results, the strong activities of proteinase in primary brewing process has been derived from the decrease of hydrogen ion concentration due to the production of organic acids.
Cyt2Aa2 is a mosquito-larvicidal protein produced as a 29 kDa crystalline protoxin from Bacillus thuringiensis subsp. darmstadiensis. To become an active toxin, proteolytic processing is required to remove amino acids from its N- and C-termini. This study aims to investigate the functional role of amino acid residues on the N-terminal ${\beta}1$ and C-terminal ${\alpha}F$ of Cyt2Aa2 protoxin. Mutant protoxins were constructed, characterized and compared to the wild type Cyt2Aa2. Protein expression data and SDS-PAGE analysis revealed that substitution at leucine-33 (L33) of ${\beta}1$ has a critical effect on dimer formation and structural stability against proteases. In addition, amino acids N230 and I233-F237 around the C-terminus ${\alpha}F$ demonstrated a crucial role in protecting the protoxin from proteolytic digestion. These results suggested that ${\beta}1$ and ${\alpha}F$ on the Nand C-terminal ends of Cyt2Aa2 protoxin play an important role in the molecular interaction and in maintaining the structural stability of the protoxin.
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