scholarly journals Influence of hydrolysis rate of urea on ruminal bacterial diversity level and cellulolytic bacteria abundance in vitro

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5475 ◽  
Author(s):  
Pengpeng Wang ◽  
Shengguo Zhao ◽  
Xuemei Nan ◽  
Di Jin ◽  
Jiaqi Wang
Keyword(s):  
Bacterial Diversity ◽  
Early Period ◽  
Urea Hydrolysis ◽  
Hydrolysis Rate ◽  
Acetohydroxamic Acid ◽  
Urease Inhibitor ◽  
Cellulolytic Bacteria ◽  
Total Bacteria ◽  
Ammonia Formation

The objective of this experiment was to evaluate the effects of urea hydrolysis rate on ruminal bacterial diversity level and cellulolytic bacteria abundance in vitro. To control urea hydrolysis rate, urea and urease inhibitor (acetohydroxamic acid, AHA) were supplemented to a 2 × 2 factorial design, with urea supplemented at 0 or 20 g/kg dry matter (DM) of substrate, and AHA equivalent to 0 or 450 mg/kg DM of substrate. Ruminal fluid was collected from three Chinese Holstein dairy cows, fed a TMR, and incubated at 39 °C for 12 h after the addition of urea and AHA. Rumen fermentation parameters, which indicated the rate of ammonia formation (including ammonia-nitrogen (NH3-N) and urea-nitrogen concentrations, urease activity, and microbial crude protein) were measured by chemical analysis. Bacterial diversity was analyzed by denaturing gradient gel electrophoresis (DGGE). Total bacteria and cellulolytic bacteria abundance was detected by quantitative PCR. Results showed that AHA addition significantly decreased the rate of ammonia formation when urea was supplemented. Urea and AHA supplementation significantly increased the bacterial community diversity level according to the Shannon–Weiner index of 16S DGGE images. Furthermore, ruminal bacterial profiles were separated by ammonia release rate when urea was supplemented, according to the DGGE and hierarchical cluster analysis. Urea supplementation reduced the abundance of cellulolytic bacteria, such asRuminococcus albus,R. flavefaciens,Fibrobacter succinogenes, andButyrivibrio fibrosolvens, but inhibition of urea hydrolysis by AHA addition alleviated the reductions during the early period of incubation. In conclusion, slow release of ammonia induced by urease inhibitor influenced the ruminal bacterial diversity level and lessened the inhibition of total bacteria growth at the incubation of 12 h andF. succinogenesduring the early period of incubation.

Kinetics and thermodynamics of urea hydrolysis in the presence of urease and nitrification inhibitors

2020 ◽  
pp. 1-11
Author(s):  
Ahmed A. Lasisi ◽  
Olalekan O. Akinremi
Keyword(s):  
Soil Temperature ◽  
Half Life ◽  
Nitrification Inhibitor ◽  
Inhibitory Effect ◽  
Urea Hydrolysis ◽  
Hydrolysis Rate ◽  
Effect Of Temperature ◽  
Nitrification Inhibitors ◽  
Urease Inhibitor ◽  
N Losses

Urease inhibitor [N-(n-butyl) thiophosphoric triamide (NBPT)] and nitrification inhibitor (NI) (3,4-dimethylpyrazole phosphate) have been used to reduce nitrogen (N) losses from urea-based fertilizers. This study evaluated the effect of temperature, NBPT, and NI on kinetic and thermodynamic properties of urea hydrolysis in six soils. Soils were amended (250 kg N·ha−1) with urea (UR), NBPT treated urea (URNBPT), or NBPT + NI treated urea (URDI), incubated at 5, 15, or 25 °C, and destructively sampled eight times during an 18 d incubation. We measured urea hydrolysis rate by the disappearance of urea with time and determined the rate constant (k; d−1) assuming first-order kinetics. Our results showed that k increased with temperature in the order of 0.07 (5 °C), 0.12 (15 °C), and 0.20 (25 °C) across soils and inhibitor treatments. In addition, k declined in the order of UR (0.19) > URDI (0.11) > URNBPT (0.08) across soils and temperatures. Although urease inhibitor, NBPT, increased the half-life of urea from 3.8 to 8.3 d across soil–temperature, the addition of a NI significantly reduced the half-life of NBPT treated urea by approximately 2 d across soil–temperature. Thermodynamics parameters showed that urea hydrolysis was nonspontaneous, and enthalpy and entropy changes were not significantly different among inhibitor treatments in five of the six soils. We conclude that the often-reported greater ammonia volatilization from URDI than URNBPT may not only be due to the persistence of ammonium in the presence of NI but also because NI reduced the inhibitory effect of NBPT on urea hydrolysis.

Intestinal Ammonia in Uraemia: The Effect of a Urease Inhibitor, Acetohydroxamic Acid

1972 ◽  
Vol 42 (1) ◽  
pp. 105-112 ◽  
Author(s):  
J. D. Swales ◽  
J. D. Tange ◽  
D. J. Evans
Keyword(s):  
Treated Group ◽  
Ammonia Concentration ◽  
Total Serum Protein ◽  
Total Serum ◽  
Acetohydroxamic Acid ◽  
Urease Inhibitor ◽  
Ammonia Production ◽  
Ammonia Content

1. The effects of intestinal ammonia in uraemia have been studied by administering a urease inhibitor, acetohydroxamic acid, to bilaterally nephrectomized rats. 2. Ammonia concentration was significantly decreased in the colon although it remained slightly higher than in normal animals. Caecal ammonia concentration was not altered. Total ammonia content was decreased in both colon and caecum. 3. A study in vitro indicated that ammonia production could be totally inhibited by acetohydroxamic acid. This suggests that the failure to suppress ammonia production in vivo is due to failure of the inhibitor to reach the site of urease production in sufficient concentration. 4. The animals treated with high doses of acetohydroxamic acid survived for a shorter time. Blood urea was higher and total serum protein lower in the treated group. The incidence of colitis was not affected.

In-vitro Kinetic Hydrolysis Study of Metronidazole Derivatives with Carvacrol and Eugenol Using Validated RP-HPLC Method

2020 ◽  
Vol 16 ◽  
Author(s):  
M. Alarjah
Keyword(s):  
Half Life ◽  
Hplc Method ◽  
Hydrolysis Rate ◽  
First Order ◽  
First Order Kinetics ◽  
Rp Hplc ◽  
Pharmacokinetic Properties ◽  
Pseudo First Order ◽  
Kinetic Hydrolysis

Background: Prodrugs principle is widely used to improve the pharmacological and pharmacokinetic properties of some active drugs. Much effort was made to develop metronidazole prodrugs to enhance antibacterial activity and or to improve pharmacokinetic properties of the molecule or to lower the adverse effects of metronidazole. Objective: In this work, the pharmacokinetic properties of some of monoterpenes and eugenol pro metronidazole molecules that were developed earlier were evaluated in-vitro. The kinetic hydrolysis rate constants and half-life time estimation of the new metronidazole derivatives were calculated using the validated RP-HPLC method. Method: Chromatographic analysis was done using Zorbbax Eclipse eXtra Dense Bonding (XDB)-C18 column of dimensions (250 mm, 4.6 mm, 5 μm), at ambient column temperature. The mobile phase was a mixture of sodium dihydrogen phosphate buffer of pH 4.5 and methanol in gradient elution, at 1ml/min flow rate. The method was fully validated according to the International Council for Harmonization (ICH) guidelines. The hydrolysis process carried out in an acidic buffer pH 1.2 and in an alkaline buffer pH 7.4 in a thermostatic bath at 37ºC. Results: The results followed pseudo-first-order kinetics. All metronidazole prodrugs were stable in the acidic pH, while they were hydrolysed in the alkaline buffer within a few hours (6-8 hr). The rate constant and half-life values were calculated, and their values were found to be 0.082- 0.117 hr-1 and 5.9- 8.5 hr., respectively. Conclusion: The developed method was accurate, sensitive, and selective for the prodrugs. For most of the prodrugs, the hydrolysis followed pseudo-first-order kinetics; the method might be utilised to conduct an in-vivo study for the metronidazole derivatives with monoterpenes and eugenol.

scholarly journals Nociceptive Sensitization by Activation of Protease-Activated Receptor 2 in a Rat Model of Incisional Pain

2021 ◽  
Vol 11 (2) ◽  
pp. 144
Author(s):  
Kanta Kido ◽  
Norika Katagiri ◽  
Hiromasa Kawana ◽  
Shigekazu Sugino ◽  
Masanori Yamauchi ◽  
...  
Keyword(s):  
Postoperative Pain ◽  
Inflammatory Responses ◽  
Early Period ◽  
Intraplantar Injection ◽  
C Fibers ◽  
Nociceptive Responses ◽  
Incisional Pain ◽  
Protease Activated Receptor 2

Postoperative pain and consequent inflammatory responses after tissue incision adversely affects many surgical patients due to complicated mechanisms. In this study, we examined whether activation of protease-activated receptor 2 (PAR-2), which is stimulated by tryptase from mast cells, elicits nociception and whether the PAR-2 antagonist could reduce incisional nociceptive responses in vivo and in vitro. The effects of a selective PAR-2 antagonist, N3-methylbutyryl-N-6-aminohexanoyl-piperazine (ENMD-1068), pretreatment on pain behaviors were assessed after plantar incision in rats. The effects of a PAR-2 agonist, SLIGRL-NH2, on nociception was assessed after the injection into the hind paw. Furthermore, the responses of C-mechanosensitive nociceptors to the PAR-2 agonist were observed using an in vitro skin–nerve preparation as well. Intraplantar injection of SLIGRL-NH2 elicited spontaneous nociceptive behavior and hyperalgesia. Local administration of ENMD-1068 suppressed guarding behaviors, mechanical and heat hyperalgesia only within the first few hours after incision. SLIGRL-NH2 caused ongoing activity in 47% of C-mechanonociceptors in vitro. This study suggests that PAR-2 may support early nociception after incision by direct or indirect sensitization of C-fibers in rats. Moreover, PAR-2 may play a regulatory role in the early period of postoperative pain together with other co-factors to that contribute to postoperative pain.

scholarly journals Effects of Methanogenic Inhibitors on Methane Production and Abundances of Methanogens and Cellulolytic Bacteria inIn VitroRuminal Cultures

2011 ◽  
Vol 77 (8) ◽  
pp. 2634-2639 ◽  
Author(s):  
Zhenming Zhou ◽  
Qingxiang Meng ◽  
Zhongtang Yu
Keyword(s):  
Methane Production ◽  
Volatile Fatty Acids ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Cellulolytic Bacteria ◽  
Bacterial Populations ◽  
Content Type ◽  
Specific Pcr ◽  
Methanogen Population ◽  
Propynoic Acid

ABSTRACTThe objective of this study was to systematically evaluate and compare the effects of select antimethanogen compounds on methane production, feed digestion and fermentation, and populations of ruminal bacteria and methanogens usingin vitrocultures. Seven compounds, including 2-bromoethanesulphonate (BES), propynoic acid (PA), nitroethane (NE), ethyltrans-2-butenoate (ETB), 2-nitroethanol (2NEOH), sodium nitrate (SN), and ethyl-2-butynote (EB), were tested at a final concentration of 12 mM. Ground alfalfa hay was included as the only substrate to simulate daily forage intake. Compared to no-inhibitor controls, PA, 2NEOH, and SN greatly reduced the production of methane (70 to 99%), volatile fatty acids (VFAs; 46 to 66%), acetate (30 to 60%), and propionate (79 to 82%), with 2NEOH reducing the most. EB reduced methane production by 23% without a significant effect on total VFAs, acetate, or propionate. BES significantly reduced the propionate concentration but not the production of methane, total VFAs, or acetate. ETB or NE had no significant effect on any of the above-mentioned measurements. Specific quantitative-PCR (qPCR) assays showed that none of the inhibitors significantly affected total bacterial populations but that they did reduce theFibrobacter succinogenespopulation. SN reduced theRuminococcus albuspopulation, while PA and 2NEOH increased the populations of bothR. albusandRuminococcus flavefaciens. Archaeon-specific PCR-denaturing gradient gel electrophoresis (DGGE) showed that all the inhibitors affected the methanogen population structure, while archaeon-specific qPCR revealed a significant decrease in methanogen population in all treatments. These results showed that EB, ETB, NE, and BES can effectively reduce the total population of methanogens but that they reduce methane production to a lesser extent. The results may guide futureinvivostudies to develop effective mitigation of methane emission from ruminants.

Skate-Skin Mucin, Rich in Sulfated Sugars and Threonine, Promotes Proliferation of Akkermansia muciniphila in Feeding Tests in Rats and In Vitro Fermentation Using Human Feces

2022 ◽  
Author(s):  
Takaaki Miyata ◽  
Takayasu Mizushima ◽  
Nobuyuki Miyamoto ◽  
Takahiro Yamada ◽  
Koji Hase ◽  
...  
Keyword(s):  
Metabolic Disorders ◽  
Control Diet ◽  
Inverse Correlation ◽  
Dietary Factors ◽  
Amino Sugars ◽  
Human Feces ◽  
In Vitro Fermentation ◽  
Akkermansia Muciniphila ◽  
Total Bacteria

Abstract Dietary factors, affect Akkermansia muciniphila (AM) abundance in the colon, have attracted attention, driven by the inverse correlation between AM abundance and metabolic disorders. We prepared skate-skin mucin (SM), porcine stomach mucin (PM), and rat gastrointestinal mucin (RM). SM contained more sulfated sugars and threonine than PM or RM. Rats were fed a control diet or diets including SM, PM, or RM (15 g/kg), or SM (12 g/kg) from five different threonine contents for 14 d. Cecal total bacteria and AM were less and more numerous, respectively, in SM-fed rats than the others, but SM did not affect microbial species-richness. Low-threonine SM did not induce AM proliferation. The in vitro fermentation with human feces showed that the rate of AM increase was greater with SM than PM. Collectively, heavy SM sulfation facilitates a priority supply of SM-derived amino sugars and threonine that promotes AM proliferation in rats and human feces.

scholarly journals Clumps of Mesenchymal Stem Cells/Extracellular Matrix Complexes Generated with Xeno-Free Chondro-Inductive Medium induce Bone Regeneration via Endochondral Ossification

Biomedicines ◽  
2021 ◽  
Vol 9 (10) ◽  
pp. 1408
Author(s):  
Susumu Horikoshi ◽  
Mikihito Kajiya ◽  
Souta Motoike ◽  
Mai Yoshino ◽  
Shin Morimoto ◽  
...  
Keyword(s):  
Stem Cells ◽  
Extracellular Matrix ◽  
Mesenchymal Stem Cells ◽  
Bone Formation ◽  
Bone Regeneration ◽  
Endochondral Ossification ◽  
Healing Process ◽  
Early Period ◽  
Cartilage Matrix

Three-dimensional clumps of mesenchymal stem cells (MSCs)/extracellular matrix (ECM) complexes (C-MSCs) can be transplanted into tissue defect site with no artificial scaffold. Importantly, most bone formation in the developing process or fracture healing proceeds via endochondral ossification. Accordingly, this present study investigated whether C-MSCs generated with chondro-inductive medium (CIM) can induce successful bone regeneration and assessed its healing process. Human bone marrow-derived MSCs were cultured with xeno-free/serum-free (XF) growth medium. To obtain C-MSCs, confluent cells that had formed on the cellular sheet were scratched using a micropipette tip and then torn off. The sheet was rolled to make a round clump of cells. The cell clumps, i.e., C-MSCs, were maintained in XF-CIM. C-MSCs generated with XF-CIM showed enlarged round cells, cartilage matrix, and hypertrophic chondrocytes genes elevation in vitro. Transplantation of C-MSCs generated with XF-CIM induced successful bone regeneration in the SCID mouse calvaria defect model. Immunofluorescence staining for human-specific vimentin demonstrated that donor human and host mouse cells cooperatively contributed the bone formation. Besides, the replacement of the cartilage matrix into bone was observed in the early period. These findings suggested that cartilaginous C-MSCs generated with XF-CIM can induce bone regeneration via endochondral ossification.

scholarly journals Profiles of Odd- and Branched-Chain Fatty Acids and Their Correlations With Rumen Fermentation Parameters, Microbial Protein Synthesis and Bacterial Populations Based on Pure Carbohydrate Incubation in Vitro

2020 ◽  
Author(s):  
Hangshu Xin ◽  
Xin Liu ◽  
Xin Jiang ◽  
Chunlong Liu ◽  
Shuzhi Zhang ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Volatile Fatty Acids ◽  
Cellulolytic Bacteria ◽  
Bacterial Populations ◽  
Branched Chain Fatty Acids ◽  
Degrading Bacteria ◽  
Branched Chain ◽  
Fermentation Parameters ◽  
Chain Fatty Acids

Abstract Background: The objectives of this study were to evaluate the profiles of odd- and branched-chain fatty acids (OBCFA; including C15:0, iso-C15:0, anteiso-C15:0, iso-C16:0, C17:0, iso-C17:0 and anteiso-C17:0) during pure carbohydrates incubation in vitro and whether they correlated with ruminal fermentation parameters, microbial crude protein (MCP) synthesis, and bacterial populations. The pure substrates containing five different ratios of fiber and starch (F:S; 0:100, 25:75, 50:50, 75:25 and 100:0) were incubated for 6 h, 12 h, 18 h and 24 h. Results: Except iso-C17:0, OBCFA concentrations were interacted by F:S and incubation time. The highest concentration of total OBCFA was found in the fermented mixture after 24 h of incubation when the F:S = 0:100; while the lowest level was 1.65 mg/g DM produced after 6 h of incubation with F:S = 50:50. The concentrations of total volatile fatty acids (TVFA) and MCP remarkably decreased linearly as the inclusion of fiber in the substrates increased, as expected. The proportions of investigated cellulolytic bacteria in our study were increased linearly (or linearly and quadratically) while those of R. amylophilus and S. bovis were decreased as fiber inclusion increased. The correlation analysis indicated that iso-C16:0 concentration might have potential as a marker of productions of TVFA and MCP with ρ being 0.78 and 0.82 respectively. Compared to starch degrading bacteria, cellulolytic bacteria had more correlations with OBCFA profiles, and the strongest association was found on the population of R. flavefaciens with C15:0 concentration (ρ = 0.70). Conclusions: Our study shows there might be scope for iso-C16:0 to predict rumen productions of VFA and MCP. Notedly, this is the first paper reporting linkage of OBCFA with rumen function based on pure carbohydrate in vitro incubation, which would avoid confounding interference from dietary protein and fat presence. However, more in-depth experiments are needed to substantiate the current findings.

scholarly journals Microbial proportion and heterotroph CO2 flux from drainage peatland under oil palm plantation

2021 ◽  
Vol 9 (1) ◽  
pp. 3055-3061
Author(s):  
Eka Lupitasari ◽  
Rahayu Widyastuti ◽  
Heru Bagus Pulunggono
Keyword(s):  
Oil Palm ◽  
Water Movement ◽  
Soil Layer ◽  
Co2 Flux ◽  
Cellulolytic Bacteria ◽  
Microbial Decomposition ◽  
Oil Palm Plantation ◽  
Ligninolytic Fungi ◽  
The Difference ◽  
Total Bacteria

The difference in soil layer can affect heterotroph respiration that means CO2 fluxes from microbial decomposition in peatlands. Oil palm plants release root exudates transported to other places, i.e., shrub, by water movement, which can stimulate microbial activity. This study was conducted to learn the effects of differences of the soil layer and distance from the trunk in drainage peatland under oil palm plantation on total bacteria, fungi, cellulolytic bacteria, ligninolytic fungi, and heterotroph fluxes CO2, then compared to a shrub. Heterotroph respiration decreased with soil layer depth, where at the layer 0-20 cm released amount of CO2 as much 6.07 + 1.76, at 20-40 cm was 5.18 + 0.50, and at 40-60 cm 5.27 + 1.20 mg CO2 100 g-1 day-1, and tended higher than in shrub where a layer of 0-20 cm released 5.51 + 1.69, then decrease at 20-40 cm to 4.83 + 1.38, and at 40-60 cm 4.30 + 1.08 mg CO2 100 g-1 day-1. Total bacteria (107 CFU g-1) and fungi (105 CFU g-1) were higher than total cellulolytic bacteria (103 CFU g-1) and ligninolytic fungi (102 CFU g-1) in both under oil palm plantation and shrub. Organic acids affected the abundance of total bacteria and fungi but did not affect cellulolytic bacteria and ligninolytic fungi on both sites, as shown by a lower population and low cellulose and laccase enzymes. These findings showed that heterotroph CO2 flux tended higher in oil palm plantations and lignocellulolytic microbes are not the only source of heterotroph respiration.

Structural and Functional Properties of Slowly Digestible Starch from Chinese Chestnut

2018 ◽  
Vol 14 (3) ◽  
Author(s):  
Xiaoyu Wen ◽  
Yuan Zhao ◽  
Joe M. Regenstein ◽  
Fengjun Wang
Keyword(s):  
Functional Properties ◽  
Profile Analysis ◽  
Hydrolysis Rate ◽  
Modified Starch ◽  
Scanning Calorimetry ◽  
Structural And Functional Properties ◽  
Modified Starches ◽  
Chestnut Starch ◽  
Slowly Digestible Starch

Abstract The structural and functional properties of slowly digestible starch (SDS) modified using pullulanase and prepared from chestnut starch were studied. The modified chestnut starches had 41.9 % SDS, which was higher than native chestnut starches (6.51 %) and cooked chestnut starches (18.6 %). The hydrolysis rate of the modified starches was 74.1 %. Scanning electron microscopy showed that the modified starch granules had a large surface area with signs of cracks and dents, and the cross-sections showed hollow internal structures. X-ray diffraction indicated that the crystallisation of the starch changed from the Cb-type to the V-type, although it retained a few C-type characteristics. Compared with native chestnut starch, the modified starches have a higher gelatinisation temperature using differential scanning calorimetry; and the texture profile analysis hardness, chewiness, cohesiveness, and gumminess of modified starch gels decreased significantly, while adhesiveness increased. When debranched using pullulanase there was a decreased solubility, swelling power, and freeze-thaw stability of the modified starches. These findings suggest that pullulanase modification changed the in vitro digestibility and crystalline structure of the modified starches.

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