In order to determine the appropriate sampling time for blood metabolites of periparturient cows, the changes of the blood composition in relation to time of day were evaluated in sixteen multiparous Holstein cows at 1 wk prepartum, 1 and 6 d postpartum. Blood samples were collected at 08:30, 10:00, 15:30 and 17:00 h in each sampling day, and the sampling times at 08:30 and 15:30 h were prior to feeding. The rectal temperature of cows increased gradually from 08:30, to 17:00 h, but blood Hct and Hb decreased constantly. Plasma non-esterified fatty acid (NEFA) concentration at 08:30 h was two-fold higher than those at 10:00, 15:30 and 17:00 h from 1 wk prepartum to 6 d postpartum, and the value was maximum at 1 d postpartum. The highest plasma urea-N was observed at 10:00 h from 1 wk prepartum to 6 d postpartum. Plasma glucose and total protein were not affected by sampling time. The data indicated that blood samples of periparturient cows should be collected before morning feeding for the diagnosis of energy status, because plasma NEFA was the highest before morning feeding.
In order to assay the efficacy of newly synthesized antiviral compounds, in vitro studies of their active intracellular phosphorylated metabolites were established as compared with Zidovudine (ZDV). Antiviral base analogs require intracellular phosphorylation prior to the inhibition of HIV replication. Therefore, antiviral drugs concentrations in plasma have not reflected any direct relationship with activity or toxicity. A method has been developed to measure the concentration of total phosphorylated metabolites inside peripheral blood mononuclear cells using modified commercial radioimmunoassay (RIA). ZDV 5'-monophosphate was synthesized and used as a procedural control for RIA modification. PBMCs were isolated from whole blood and incubated with ZDV for 20 h to allow metabolic phosphorylation. Viable cells were extracted overnight with 60% methanol. After evaporation, the extract was reconstituted in Tris buffer. Samples were split into two fractions, one of which was treated with alkaline phosphatase (AP) to liberate phosphate groups. Concentrations of phosphorylated metabolites were determined by subtracting thε concentration of non-AP-treated fraction from that of the treated fraction. Recovery of phosphorylated ZDV from cell extracts was approximately 90%, and reproducibility was acceptable (coefficients of variation <15% for concentrations${\geq}$0.25 ng/mL). Intracellular concentrations $(0.135{\sim}5.019\;nmole/10^6\;cells)$ followed a nonlinear dose-response relationship over the range $0.015{\sim}2.996mM$ extracellular ZDV, with concentration-dependant saturation.
An analytical method the determination of benzo(a)pyrene (BaP) and its hydroxylated metabolites, 1-hydroxybenzo(a)pyrene (1-OHBaP), 3-hydroxybenzo(a)pyrene (3-OHBaP), benzo(a)pyrene-4,5-dihydrodiol (4,5-diolBaP) and benzo(a)pyrene-7,8-dihydrodiol (7,8-diolBaP), in rat urine and plasma has been developed by HPLC/FLD and GC/MS. The derivatization with alkyl iodide was employed to improve the resolution and the detection of two mono hydroxylated metabolites, 1-OHBaP and 3-OHBaP, in LC and GC. BaP and its four metabolites in spiked urine were successfully separated by gradient elution on reverse phase ODS $C_{18}$ column (4.6 mm I.D., 100 mm length, particle size 5 ㎛) using a binary mixture of MeOH/H₂O (85/15, v/v) as mobile phase after ethylation at 90 ℃ for 10 min. The extraction recoveries of BaP and its metabolites in spiked samples with liquid-liquid extraction, which was better than solid phase extraction, were in the range of 90.3- 101.6% in n-hexane for urine and 95.7-106.3% in acetone for plasma, respectively. The calibration curves has shown good linearity with the correlation coefficients (R²) varying from 0.992 to 1.000 for urine and from 0.996 to 1.000 for plasma, respectively. The detection limits of all analytes were obtained in the range of 0.01-0.1 ng/mL for urine and 0.1-0.4 ng/mL for plasma, respectively. The metabolites of BaP were excreted as mono hydroxy and dihydrodiol forms after intraperitoneal injection of 20 mg/kg of BaP to rats. The total amounts of BaP and four metabolites excreted in dosed rat urine were 3.79 ng over the 0-96 hr period from adminstration and the excretional recovery was less than 0.065% of the injection amounts of BaP. The proposed method was successfully applied to the determination of BaP and its hydroxylated metabolites in rat urine and plasma for the pharmacokinetic studies.
This study was carried out to evaluate the effects of fractions of methanol(MeOH) extracts of Lycopus lucidic Turcz on hyperglycemia and energy metabolites in streptozotocin(STZ) diabetic rats. Diabetes mellitus was induced in male Sprague-Dawley rats weighing 200-220 g by an injection of STZ dissolved in a citrate buffer into the tail vein at a dose of 45 mg/kg of body weight, and the rats were divided into 7 groups, that is, one normal group and 6 diabetic groups: STZ-control, hexane, chloroform(CHCl$\sub$3/). ethylacetate(EtOAc), butanol(BuOH) and H$\sub$2/O fraction-fed groups. All groups were fed an AIN-93 diet and the fractions of Lycopus lucidic Turcz were administered orally with 2 % Tween 80 for 14 days after the STZ injection. Body weight, diet intake and organ weights were monitored. The plasma levels of blood glucose, insulin and protein were determined. The plasma concentrations of cholesterol, HDL-cholesterol, triglycerides and free fatty acid were assayed. The plasma activities of aspartate aminotransferase (AST) and alanine aminotransferase(ALT) were also measured. Body weight losses were observed by feeding the fractions of Lycopus lucidic Turcz in STZ experimental groups, and the kidney weight was increased. The extent of blood glucose decrement was significantly greater in the hexane and BuOH fraction-fed groups than STZ-control group. The plasma protein level was significantly lower in the H$\sub$2/O fraction-fed group. The plasma cholesterol level was decreased in BuOH and H$\sub$2/O fraction-fed groups compared with the STZ-control group. The levels of free fatty acids in the CHC1$\sub$3/ and H$\sub$2/O fraction-fed groups were significantly decreased(p<0.05). ALT activitiy of BuOH fraction-fed group was lower than control but it was not significantly different. These results suggest that the fractions of Lycopus lucidic Turcz are capable of lowering blood glucose and fat metabolites concentrations when administered to STZ-treated rats, and AST/ALT activity and insulin levels show the possibility of therapeutic use to diabetes mellitus.
Alipour, Fereshteh;Vakili, Alireza;Mesgaran, Mohsen Danesh;Ebrahimi, Hadi
Asian-Australasian Journal of Animal Sciences
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제32권11호
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pp.1695-1704
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2019
Objective: This study investigated the effects of the administration of ethanolic saffron petal extract (SPE) and vitamin E (Vit E) on growth performance, blood metabolites and anti-oxidant status in Baluchi lambs. Methods: Thirty-two Baluchi male lambs ($35.22{\pm}5.75kg$) were randomly divided into 4 groups. The 1st (control), 2nd (injectable saffron petal extract [ISPE]), and 3rd (Vit E) groups were respectively injected subcutaneously with either physiological saline (5 mL), SPE (25 mg/kg body weight [BW]) or DL-${\alpha}$-tocopheryl acetate (225 IU) once a week. An oral dose of SPE (500 mg/kg BW) was also administered to the 4th group (oral saffron petal extract [OSPE]). Feed intake and BW were measured for 42 days and blood samples were taken on days 1, 14, 28, and 42. The lambs were slaughtered, and tissue samples were taken. Results: Growth performance and many blood metabolites were not affected (p>0.05) by the treatments. Cholesterol of plasma in the ISPE and Vit E groups was similar and less (p<0.01) than both the OSPE and control groups. Although there was no significant difference between the control and other groups for plasma triglyceride, the ISPE group showed lower (p<0.05) triglyceride than the OSPE and Vit E groups. The highest (p<0.01) plasma glutathione peroxidase (GPx) was detected in the OSPE group, while the ISPE and Vit E groups showed higher (p<0.01) superoxide dismutase (SOD) of plasma than the control. Malondialdehyde of plasma in the ISPE group was lower (p<0.05) than the OSPE. No differences (p>0.05) were observed among the groups for antioxidant status of both longissimus dorsi muscle and liver. However, the activity of GPx in the kidney and heart, as well as SOD activity in the kidney, were influenced ($p{\leq}0.01$) by the treatments. Conclusion: Adding ethanolic SPE improved antioxidant status and lowered lipids oxidation in lambs. The SPE and Vit E demonstrated similar effects on antioxidant status in lambs.
The aim of this study was to identify useful secondary traits for estimating genetic ability of milk production traits. We investigated the value of using plasma metabolites concentrations. Two hundred and nineteen cattle out of 271 had only milk production traits records (G1), 33 had only metabolites records (G2), and 19 had both milk production traits and metabolites records (G3). Fifty two calves with metabolites records (G2 and G3) were born from 1992 to 1997. Forty three calves (29 females, 14 males) were used from 10 to 90 d of age and the others (3 females, 6 males) from 10 to 60 d of age. A total of 566 records of milk yield, fat yield and protein yield for 240 to 305 d on 238 heads (G1 and G2) were collected The collected blood samples were divided into three age groups: AG1, 10 to 30 d; AG2, 40 to 60 d; and AG3, 70 to 90 d. Heritabilities of milk yield, fat yield and protein yield were $0.45{\pm}0.04$, $0.50{\pm}0.04$ and $0.38{\pm}0.04$, respectively. Heritability of plasma glucose concentration at AG1 was $0.45{\pm}0.08$. Genetic correlations between plasma glucose concentration and milk yield, fat yield and protein yield were -$0.35{\pm}0.28$, $0.64{\pm}0.24$ and $0.36{\pm}0.35$, respectively. When the plasma glucose concentration at AG1 was used to estimate genetic ability of these milk production traits, reliability of milk yield of animals without milk record increased 8.2%, fat yield increased 24.2% and protein yield increased 9.5%. Heritability of plasma total cholesterol concentration at AG3 was $0.83{\pm}0.04$. Genetic correlation between plasma total cholesterol concentration and milk yield, fat yield and protein yield were $0.58{\pm}0.21$, $0.42{\pm}0.20$ and $0.45{\pm}0.22$, respectively. When the plasma total cholesterol concentration at AG3 was using to estimate genetic ability of these milk production traits, reliability of milk yield of animals without milk record increased 19.0%, fat yield increased 9.6%, and protein yield increased 13.5%. The annual genetic gain is in proportion to the reliability of selection. These results show that the plasma metabolite concentrations would be useful for improvement of genetic ability for milk production traits in the genetic improvement in herd of cows, where half of the animals selected are from a herd without its own milk record.
Vakili, A.R.;Khorrami, Behzad;Mesgaran, M. Danesh;Parand, E.
Asian-Australasian Journal of Animal Sciences
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제26권7호
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pp.935-944
/
2013
Essential oils have been shown to favorably effect in vitro ruminal fermentation, but there are few in vivo studies that have examined animal responses. The objective of this study was to evaluate the effects of thyme (THY) and cinnamon (CIN) essential oils on feed intake, growth performance, ruminal fermentation and blood metabolites in feedlot calves fed high-concentrate diets. Twelve growing Holstein calves ($213{\pm}17kg$ initial BW) were used in a completely randomized design and received their respective dietary treatments for 45 d. Treatments were: 1-control (no additive), 2-THY (5 g/d/calf) and 3-CIN (5 g/d/calf). Calves were fed ad libitum diets consisting of 15% forage and 85% concentrate, and adapted to the finishing diet by gradually increasing the concentrate ratio with feeding a series of transition diets 5 wk before the experiment started. Supplementation of THY or CIN did not affect DMI and ADG, and feed efficiency was similar between treatment groups. There were no effects of additives on ruminal pH and rumen concentrations of ammonia nitrogen and total VFA; whereas molar proportion of acetate and ratio of acetate to propionate decreased, and the molar proportion of propionate increased with THY and CIN supplementation. Rumen molar concentration of butyrate was significantly increased by adding CIN compared to control; but no change was observed with THY compared with control group. No effects of THY, or CIN were observed on valerate, isobutyrate or isovalerate proportions. Plasma concentrations of glucose, cholesterol, triglyceride, urea-N, ${\beta}$-hydroxybutyrate, alanine aminotransferase and aspartate aminotransferase were not changed by feeding THY or CIN. Results from this study suggest that supplementing a feedlot finishing diet with THY or CIN essential oil might be useful as ruminal fermentation modifiers in beef production systems, but has minor impacts on blood metabolites.
Thirty Holstein calves were used to determine effects of age, environment and sex on blood metabolite concentrations during 1 to 90 d of age. Calves were weaned at 75 d of age. Environmental effects are grouped by the difference in month at birth and site of feeding. Blood samples were obtained every 2 or 3 d. The mean metabolite concentration every 3 d was used for the statistical analysis. Dairy bodyweight gain was not affected by environmental group and sex effect. Concentrations of plasma glucose, nonesterified fatty acids (NEFA), triglyceride, total cholesterol and total ketone changed with growth. These developmental changes in metabolite levels would be caused by ruminal maturation with increment of grain intake. Levels of plasma urea nitrogen, glucose, NEFA, triglyceride and total cholesterol drastically changed during a few weeks after birth, indicating that the physiological state in calves greatly changed during that time. Effects of the environmental group and sex were significant in almost all metabolites. Temperature influenced plasma metabolite concentrations. The plasma metabolite concentrations were affected more intensely by heat stress in the infant period than in the neonatal period.
Six crossbred goats in their 2nd or 3rd lactation, were administered bromocryptine at 5 mg/day during early lactation of 15-20 days (period I) and thereafter again at an interval of 13 days, bromocryptine was given for 5 days (period II). Blood samples were collected before (-5, -4, -3, -2, -1) during (1, 2, 3, 4, 5) and after (+1, +2, +3, +4, +5) administration of bromocryptine in both the periods of study. In period I, administration of bromocryptine resulted in a decrease in milk yield to the extent of 16..8% in comparison to before treatment, and 28.5% after the cessation of treatment. The glucose content of blood increased (p<0.01) as the milk yield decreased without any change in NEFA concentration. During period II of bromocryptine treatment the milk yields did not change in spite of a decline in prolactin level, perhaps the effect of previous treatment was prolonged. A decline in protein and lactose content of milk after bromocryptine treatment in both the periods of study, when prolactin level also declined suggests a role of prolactin in protein synthesis and also a depressing effect on lactose synthesis.
Objective: The objective of the study was to determine the relationship between plasma and salivary cortisol concentrations in beef cattle that were subjected to handling prior to sampling. Methods: Twenty-one Nguni cows of three age categories; 5 to 7 yr (n = 7), 8 to 10 yr (n = 6), and 11 to 13 yr (n = 8) were handled for five consecutive weeks. In the pen, a human avoidance test was performed and cattle responses to restraint in the chute and crush were observed. In addition, rectal temperature readings were taken and, faecal samples were collected and analysed for glucocorticoid metabolites. Through the handling and restraint process, excretory and vocalisation behaviour, as a sign of stress were observed and recorded. Thereafter, six cows were randomly selected and subjected to an adrenocorticotropic hormone (ACTH) challenge. Blood and saliva samples were extracted to determine cortisol concentrations. Results: Repeated handling affected (p<0.05) faecal glucocorticoid metabolites, rectal temperatures, avoidance distance, crush scores as well as urination and defaecation behaviour. Acclimation to handling was variable based on each respective parameter. Saliva cortisol concentrations increased and decreased significantly (p<0.001). A peak value of $136.78{\pm}15.869nmol/L$ was observed 30min after administration of ACTH, from a baseline value of $8.75{\pm}15.869nmol/L$. Plasma cortisol concentrations did not differ (p>0.05) across the time of sampling. A low and insignificant correlation (r = 0.0131, p>0.05) between plasma and saliva cortisol was therefore observed. Conclusion: We conclude that if beef cows are subjected to handling prior to sampling, a weak relationship exists between plasma and salivary cortisol levels.
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