The influence of different doses of α-tocopherol and ascorbic acid on selected oxidative stress parameters in in vitro culture of leukocytes isolated from transported calves

The objective of the study was to assess in vitro the influence of various doses of two different antioxidants, α-tocopherol and ascorbic acid, on protective mechanisms against ROS in white blood cells obtained from calves exposed to transport, and to compare these results with those obtained from non-transported animals.The concentrations of lipid peroxidation products in leukocytes and in the retained medium were assessed by determining the level of ThioBarbituric Acid Reactive Substances (TBARS). Total antioxidant status in the leukocytes and the medium were estimated using a ferric-reducing ability of plasma (FRAP) assay. Leukocyte viability was determined using the trypan blue reduction test.The study demonstrated that after bovine leukocytes (WBC) were incubated in vitro with α-tocopherol and ascorbic acid, peroxidation intensity decreased and total antioxidant capacity increased. The results of the study reveal that these antioxidants in concentrations over 0.1 mg/ml have a major impact on free radical activity on bovine white blood cells and on cell viability during transport of animals.Based on this study, we suggest that incubation of the leukocytes with antioxidants decreases the oxidative stress development, which can be helpful in protection of the immunological cells during bovine respiratory disease.     

玉米γ-生育酚甲基转移酶基因的克隆与分析

[目的]对玉米γ-生育酚甲基转移酶（γ-TMT）基因进行克隆和分析研究。[方法]通过RT-PCR扩增得到玉米γ-TMT基因的cDNA序列,同时对序列进行同源性分析、氨基酸序列比对和系统进化分析。[结果]玉米γ-TMT基因cDNA全长1 310 bp,包含一个长为1 059bp的完整开放读码框,推导的氨基酸序列与小麦和水稻等高等植物的γ-TMT基因同源性较高,序列一致率为69%～87%;与低等生物褐藻的序列同源性较低,一致率只有56%。系统进化分析表明不同来源的γ-TMT基因可聚为3类。[结论]为进一步研究γ-TMT基因的功能奠定了基础。     

Conceptus-related measurements during the first trimester of bovine pregnancy

In order to determine the variability inherent in conceptus-related measurements in first trimester bovine pregnancies, conceptus and fetometric parameters from beef cattle pregnancies (n=103) estimated to be between Days 36 and 103 of gestation were examined. During this period, the protein concentration of amniotic fluid ranged between 0.181 and 0.501mg/mL. The amniotic fluid volume gradually increased from <1mL at Day 36 to 950mL at Day 103 (R(2)=0.9275) and amniotic compartment dimensions (length, R(2)=0.9713; width, R(2)=0.9802) increased predictably with fetal growth. Conversely, allantoic fluid protein concentration and volume correlated weakly with fetal age. A significant linear correlation existed between fetal crown rump length (CRL) and crown nose length (R(2)=0.9899) confirming that either measurement can be employed in the ultrasonographic estimation of fetal age. The amniotic compartment and fetometric data presented here have both research and clinical value, particularly in relation to fetal development evaluation and pregnancy viability diagnosis.     

Effect of Freezing Rates and Supplementation of α-Tocopherol in the Freezing Extender in Equine Sperm Cryosurvival

This study was conducted to determine the impact of antioxidant (α-tocopherol) supplementation in the cryopreservation extender and three different freezing rates (FRs) on quality of post-thaw semen to elaborate a new protocol for stallion semen cryopreservation. Six ejaculates from each of four stallions were subjected to cryopreservation with a commercial extender (Gent, Minitub Iberia, Spain), without any supplementation (control) or supplemented with 2 mM α-tocopherol. The semen was exposed to three different freezing rates between 5°C and −15°C: slow (5°C/min), moderate (10°C/min), and fast (20°C/min). After thawing, the sperm viability (Sybr-14 and propidium iodide [PI]), mitochondrial membrane potential (MMP) (5,5′,6,6′-tetrachloro-1,1′,3,3′tetraethylbenzimidazolyl carbocyanine iodine), lipid membrane peroxidation (LPO; C₁₁-BODIPY^581/591), and apoptosis status (fluorescein isothiocyanate–conjugated annexin V and PI) of the plasmatic membrane of each sample were determined by flow cytometry. For both extenders, the percentage of viable cells was higher for spermatozoa cooled at 5°C/min than at 10°C/min and 20°C/min (P ≤ .05). The FR of 20°C/min demonstrated a lower value of MMP than the FR of 5°C/min and 10°C/min (P ≤ .05). The α-tocopherol extender improved (P ≤ .05) post-thaw membrane LPO; however, it did not improved viability and the apoptosis status of the sperm plasmatic membrane after thawing. In conclusion, results clearly indicate that the cryosurvival of stallion spermatozoa is improved when FR of 5°C/min is used from 5°C to −15°C.     

Effect of three irrigation regimes on Arbequina olive oil produced under Tunisian growing conditions

This study investigated the effect of irrigation amount on the concentration of phenolic compounds in olive (Olea europaea L., cv. Arbequina) oil obtained from an intensively-managed orchard in a semi-arid area with a Mediterranean climate in Tunisia. Different irrigation treatments 50% Etc, 75% Etc and 100% Etc were applied to the olive orchard. Oil quality, evaluated using the parameters established to determine the quality level of virgin olive oils (acidity, K232, K270 and peroxide index) was slightly affected by irrigation. However, results showed that irrigation positively affected both fruit and oil quality. In fact, the least irrigation regime (T1), showed a significantly higher content of oleic acid (70.08%), whereas olive oils from more irrigated trees (T2 and T3) had higher contents of palmitic acid (11.64% and 13.14%, respectively) and lower of linoleic acid (approximately 12.7%). However, content of phenolic compounds (hydrophilic and lypophilic), in the oils extracted, strongly differed. In fact, different irrigation regimes applied not only affected the total amount of phenols which were proportional to irrigation (193.2 and 271.87 mg kg⁻¹ for T1 and T3, respectively) except for T2 but also their HPLC profiles. Contrarily to phenols, insignificant differences were observed in the concentration of α-tocopherol between the irrigation treatments studied.     

The Effect of Coenzyme Q10 and α-Tocopherol in Skim Milk–Based Extender for Preservation of Caspian Stallion Semen in Cool Condition

The purpose of this study was to determine the effects of different concentrations of coenzyme Q₁₀ (CoQ₁₀) and α-tocopherol (T) along with their interaction effects on the quality of preserved stallion semen at 5°C for a period of 48 hours. Semen was collected and diluted with skim milk–based extender that was supplemented with different antioxidants: no antioxidant (negative control [NC]), 0.9% (vol/vol) dimethyl sulfoxide (positive control [PC]), α-tocopherol (5 [T5] or 10 [T10] mM), CoQ₁₀ (1 [C1] or 2 [C2] μM), 1 μM CoQ₁₀ + 5 mM α-tocopherol (C1T5), 1 μM CoQ₁₀ + 10 mM α-tocopherol (C1T10), 2 μM CoQ₁₀ + 5 mM α-tocopherol (C2T5), and 2 μM CoQ₁₀ + 10 mM α-tocopherol (C2T10), then kept at 5°C. The results showed that C1 extender resulted in higher total motility (62.44 ± 3.82) and plasma membrane integrity (65.16 ± 3.63%) compared with NC after 48 hours of storage (P < .05). Different concentrations of α-tocopherol had no significant effects on sperm quality, with the exception of plasma membrane integrity, compared with NC and PC extenders (P > .05). Also, C1T5 extender improved total and progressive motility, plasma membrane integrity and functionality, and decreased lipid peroxidation compared with NC and C2T10 extenders over 48 hours of storage at 5°C (P < .05). The C1T5 extender was similar to C1 and T5 extenders in all semen parameters evaluated during storage time. In conclusion, between previously mentioned extenders, C1T5 could improve stallion sperm quality during 48 hours of storage. In the present study, none of extenders had effect on sperm quality until 24-hour storage.     

Variability in permeability and integrity of cell membrane and depletion of food reserves in neem （Azadirachta indica） seeds from trees of different age classes

We quantified cell membrane permeability(electrical conductivityEC,water soluble sugar-WSS,and amino acids-AA)and integrity(phospholipids,α-tocopherol and lipid peroxidation)along with food reserve deterioration(total proteins,total sugar,and total starch)of neem seeds collected from various mother tree age classes and stored for 65days in airtight plastic containers at ambient room temperature(35±5°C).Results show that the activities were higher in fresh seeds(EC267.56-2950.01μS/g,WSS 19.96-19.48 mg/g and AA 5.40-5.35 mg/g)and declined with increasing duration of storage period(EC153.37-195.17μS/g,WSS 3.13-4.17 mg/g and AA 4.29-4.49 mg/g after35 days and EC 144.02-161.56μS/g,WSS 2.06-2.40 mg/g and AA3.98-4.27 mg/g after 65 days of storage).Phospholipids andα-tocopherol were higher in fresh seed(0.073-0.093 OD at 710 nm and0.080-0.105 OD,respectively)and declined as storage duration increased(0.033-0.042 OD at 710 nm and 00.0010-0.0020 OD,respectively).Dead seeds showed reduced amounts of phospholipids and minimum activity ofα-tocopherol(antioxidants).The level of MDA was lower in fresh seeds(0.0066-0.0087 OD at 600-535 nm)and increased as storage duration increased(0.0248-0.0268 OD after 65 days of storage).The higher amount of MDA indicated that seeds died due to rancidity of the oil inside the seed.Neem seed cake was assessed for deterioration of food reserves(total proteins,total sugar,and total starch),concentrations of which were higher in fresh seed and declined as storage duration increased.Germination was higher in fresh seeds and after 65days,no germination was received perhaps due to deterioration of biochemicals in seeds.Patterns of seed deterioration were similar across all seed lots.     

大豆γ-生育酚的混合遗传分析与QTL定位

【目的】通过对大豆γ-生育酚进行混合遗传和QTL定位分析,了解其遗传机制,定位其主效QTL,为高γ-生育酚含量大豆品种的选育奠定基础。【方法】以栽培大豆晋豆23为母本,以山西农家品种大豆灰布支黑豆为父本杂交衍生的重组自交系作为供试群体构建遗传图谱。图谱全长2 047.6 cM,平均图距8.8 cM,包括227个SSR标记,232个标记位点。重组自交系试验群体及亲本材料分别于2011年、2012年和2015年夏季在河南省农业科学院原阳试验基地种植,冬季在海南省三亚南繁试验基地种植。田间试验采取随机区组设计,2次重复。从6个环境中每个家系选取15.00 g籽粒饱满,大小一致的大豆种子,利用高效液相色谱法定性、定量测定样品中的γ-生育酚含量。采用主基因+多基因混合遗传分离分析法,对大豆γ-生育酚含量进行混合遗传分析;采用WinQTLCart 2.5复合区间作图法,对大豆γ-生育酚含量进行QTL定位分析。【结果】主基因+多基因混合遗传分析表明,γ-生育酚受2对重叠作用主基因×加性多基因控制。遗传基因分布在双亲中。三亚试验数据检测出2对主基因间上位性效应值为0.4010—0.5169和多基因的加性效应值为0.1797—0.2146,主基因遗传率为11.27%—13.05%,多基因遗传率为82.51%—86.55%,多基因效应大于主基因效应。原阳试验数据检测到2对主基因间上位性效应值为0.9646—1.8455,主基因遗传率为39.51%—58.96%,没有检测出多基因效应。采用WinQTLCart 2.5复合区间作图（CIM）共检测到9个影响γ-生育酚的QTL,分布于A1（Chr.5）、A2（Chr.8）、C1（Chr.4）、K（Chr.9）、M（Chr.7）和G（Chr.8）6条染色体中,单个QTL的贡献率为7.29%—29.55%。qγ-G-1同时在2011年原阳、2012年三亚、2015年三亚3个环境下检测到,且均定位在G（Chr.18）染色体Satt275—Satt038标记区间0.01 cM处,解释的表型变异分别为8.97%、8.12%和7.91%。qγ-A1-1同时在2011年原阳和2015年原阳2个环境下检测到,且均定位在A1（Chr.5）染色体Satt276—Satt364标记区间0.01 cM处,解释的表型变异分别为29.54%和28.23%。qγ-G-1和qγ-A1-1 2个QTL能够稳定遗传。【结论】γ-T最适遗传模型符合MX2-Duplicate-A,即2对重叠作用主基因×加性多基因模型。其遗传同时受到基因型、环境和上位性的影响。检测到γ-T的2个稳定主效QTL,Satt275—Satt038和Satt276—Satt364是共位标记区间。