Stallion Semen Incubated with Hydrogen Peroxide Decreased DNA Fragmentation as Measured by the TUNEL Assay

Spermatozoa are highly specialized cells that are sensitive to environmental factors such as temperature and redox state. Hydrogen peroxide (H₂O₂) is a reactive oxygen species, and its presence is often associated with a decline in sperm function. The aim of this study was to evaluate the effect of H₂O₂ and heat stress on DNA damage, membrane integrity, and motility of stallion spermatozoa. Spermatozoa from three light-horse stallions were subjected to thermal stress, treatment with H₂O₂, or a combination of both. Treatments were organized using a 2 × 2 factorial design consisting of heat stress (control, 41°C) and potential oxidative stress (control, 50 μM H₂O₂) for 1 hour. The experiments were repeated independently on all stallions. Primary motility parameters were measured using a computer-assisted sperm analysis, whereas DNA damage was assessed using the TUNEL assay. To evaluate membrane integrity, an amine reactive dye was utilized. DNA and membrane integrity were simultaneously assessed in the same flow cytometry assay. Sperm incubated at 41°C were observed to have decreased motility (76.2% vs. 66.6%; P < .05) but not progressive motility (39.2% vs. 25.6%), membrane damaged (30.8% vs. 37.4%), or DNA damage (19.7% vs. 17.3%). Interestingly, when compared to control, treatments with H₂O₂ had decreased DNA damage (19.7% vs. 7.1%; P < .05), but did not affect any other parameter. Although our experiment demonstrated a favorable effect of 50 μM H₂O₂ on DNA damage, further studies need to be conducted to confirm these findings and to clarify any possible signaling mechanisms.     

Influence of varying dietary electrolyte balance on broiler performance under tropical summer conditions

A 6‐week feeding trial was conducted to determine the effects of varying dietary electrolyte balance (DEB) on growth performance and physiological responses in broiler chicks reared during hot summer months (26.1–37.5 °C). The average minimum and maximum room temperatures recorded from 14 to 42 days of age were 26.1 and 37.5 °C, respectively, with relative humidity ranging from 51% to 55%. Five DEB treatments supplying 0, 50, 150, 250 and 350 (Na⁺+K⁺–Cl^?) mEq/kg of diet were prepared by adding NaHCO₃ and/or NH₄Cl to basal diet that contained 185 mEq/kg from 1 to 28 days and 172 mEq/kg from 29 to 42 days. These DEB diets were randomly offered to six experimental units of 20 chicks each. Improved 42‐day body weight gain (BWG), feed:gain (F:G) and reduced mortality was noted with DEB 50, 150 and 250 mEq/kg compared to DEB 0 and 350 mEq/kg. A quadratic effect of increasing DEB level was observed on BWG and F:G, whereas the effect was linear for feed intake, water intake and litter moisture. The DEB 50, 150 and 250 maintained better blood pH, pCO₂ and HCO₃ concentrations during acute heat stress periods than that of 0 and 350. The blood heterophil and lymphocyte percent as well as heterophil:lymphocyte were remained unaffected. The lowest DEB (0 mEq/kg) diet reduced the feed intake and promoted metabolic acidosis. At 350 mEq/kg DEB, water intake and metabolic alkalosis were promoted probably due to high levels of dietary NaHCO₃. The study concluded that overall better performance was recorded with DEB 50, 150 and 250 mEq/kg. These results indicated that single optimal DEB value could not be recommended to combat heat stress in broilers.     

一氧化氮信号途径参与草地早熟禾耐镉机制的研究

为探究信号分子一氧化氮（Nitric Oxide,NO）对镉（Cadmium,Cd）胁迫下草地早熟禾（Poa pratensis）的调控机理,揭示草地早熟禾耐镉机理,本试验采取根施1000 μmol·L^-1氯化镉（Cadmium chloride hemidihydrate,CdCl₂·2.5H₂O）及叶面喷施NO合成酶抑制剂（L-硝基精氨酸甲酯（L-NAME））、硝酸还原酶抑制剂（钨酸钠（Tungstate））、NO清除剂（4-羧基苯-4,4,5,5-四甲基咪唑-1-氧-3-氧化物（cPTIO））处理草地早熟禾叶片,研究不同处理对草地早熟禾幼苗叶片生长及抗氧化酶活性、光合色素含量、丙二醛（Malondialdehyde,MDA）、游离脯氨酸、内源NO含量的影响。研究发现,镉胁迫下草地早熟禾内源NO含量增加,添加一氧化氮清除剂、一氧化氮合酶抑制剂、硝酸还原酶抑制剂可增加超氧化物歧化酶（Superoxide Dismutase,SOD）、过氧化氢酶（Catalase,CAT）、抗坏血酸过氧化物酶（Ascorbate peroxidase,APX）活性,降低过氧化物酶（Peroxidase,POD）活性,增加叶绿素a、叶绿素b、类胡萝卜素、丙二醛含量,降低脯氨酸（Proline,Pro）含量、叶片相对含水量（Relative Water Content,RWC）和干物质量（Dry matter content）。结果表明,一氧化氮合酶途径可能是早熟禾合成NO的主要途径,内源NO可通过调节抗氧化酶活性、光合色素含量、游离脯氨酸含量等途径缓解镉胁迫。     

奶牛氧化应激研究进展

高产奶牛更容易发生氧化应激,氧化应激可以导致生物学大分子和生物膜系统的破坏,并可以增加炎症反应时的组织损伤：奶牛氧化应激的产生除了与机体的生理状态、健康状况和饲料成分有关外,环境因素也会引起氧化应激。目前主要通过饲喂抗氧化添加剂来预防奶牛氧化应激。     

盐胁迫对5份无芒雀麦苗期生长和生理生化的影响及综合性评价

为了明确不同无芒雀麦苗期耐盐能力的强弱,本研究选择5份不同株型的无芒雀麦作为供试材料。根据新疆土壤盐碱成分设置中性盐(M盐)：NaCl∶Na₂SO₄∶Na₂CO₃=1∶4∶0和碱性盐(A盐)：NaCl∶Na₂SO₄∶Na₂CO₃=1∶1∶8两种处理,于三叶期每盆一次性浇灌2 L电导率(EC)为20 ms·cm^-1的盐处理液,CK浇灌等量自来水。处理30 d后测定生长指标、叶绿素荧光特性参数、丙二醛(MDA)和渗透调节物质含量,并通过相关性分析、主成分分析、隶属函数分析对13个指标进行分析评价。结果表明：除F_v/F_m和F_v/F_o外,盐胁迫显著影响其余11个指标,且5份材料间差异显著(P<0.05);5份无芒雀麦的株高、茎粗、分蘖数、地上生物量、根冠比、MDA和脯氨酸含量在不同盐胁迫下表现出不同的适应变...     

干旱对草坪草的影响

综述了干旱条件下草坪草的生态适应性,细胞膜、细胞壁、叶绿体超微结构的变化,草坪草对干旱胁迫的生理生化反应,以及提高草坪抗旱性的途径。     

油菜素内酯对干旱胁迫下羊草渗透调节及抗氧化酶的影响研究

受全球气候变化影响,我国牧区干旱现象日益严重,极大限制了草原生产力的提高。本文采用盆栽人工控水的方法,研究不同浓度油菜素内酯(BR)处理对羊草生长和抗旱特性的影响。结果表明干旱显著抑制植株的生长,而BR能有效减缓干旱胁迫对羊草造成的伤害。经过不同浓度的BR处理后,与干旱胁迫相比,株高、叶面积、干重、含水量、叶绿素a、叶绿素b、类胡萝卜素、根系活力、脯氨酸、可溶性蛋白、可溶性糖和5种抗氧化酶活性均随浓度增加而升高,但高浓度下又有降低的趋势,而丙二醛(MDA)和叶片电导率则相反,其中以BR浓度为0.1 mg/L时的抗旱效果最好。研究发现适当浓度的BR提高羊草株高、叶面积和光合色素含量,促进干物质积累和根系活力,与其降低膜脂过氧化产物MDA与质膜透性,提高脯氨酸、可溶性蛋白、可溶性糖等渗透调节物质含量,增强超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)等抗氧化酶活性密切相关。     

Relationship of climatic conditions to fecal corticosterone levels of captive cheetahs reared in Japan

To assess the stress level of cheetahs reared in Japan and to identify the prime components of the climatic conditions that affect their thermal stress, fecal corticosterone was monitored for 8 months from May to the following January. A total of 203 fecal samples were gathered in the morning from seven adult cheetahs that were kept at a zoological garden in Wakayama, Japan. Cheetahs were on exhibit singly or together with a harmonious conspecific during the day, but housed singly at night. Although the monthly fluctuation in corticosterone concentrations was not significant, the concentrations were relatively low during the summer season. Individual differences among cheetahs and the interaction effect between individual and month on the corticosterone concentrations were significant. Whereas the corticosterone concentrations negatively correlated with air temperature, they were positively correlated with the amount of rainfall. The highest air temperature and the amount of rainfall were extracted as the prime factors affecting corticosterone concentrations. These results suggest that cheetahs reared in Japan are somewhat subjected to thermal stress, particularly on cooler and/or rainy days.     

Oxidative stress inducers potentiate 2,3,7,8-tetrachlorodibenzo-p-dioxin-mediated pre-cardiac edema in larval zebrafish

We reported the involvement of oxidative stress and prostaglandins including thromboxane and prostacyclin in pre-cardiac edema (early edema) caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). While the involvement of oxidative stress in TCDD-induced toxicity has been frequently reported, the mechanism of its action is still unclear. In the present study, oxidative stress inducers including paraquat, hydrogen peroxide (H₂O₂) and rotenone augmented early edema (edema) induced by a low concentration of TCDD (0.1 ppb) at 55 hr post fertilization (hpf), while each of them alone did not cause edema. Edema caused by TCDD plus oxidative stress inducers was almost abolished by antioxidants, an antagonist for thromboxane receptor (ICI-192,605) and an agonist for prostacyclin receptor (beraprost), suggesting that the site of action of these inducers was in the regular signaling pathway after activation of aryl hydrocarbon receptor type 2 (AHR2) by TCDD. Oxidative stress inducers also enhanced edema caused by an agonist for the thromboxane receptor (U46619), and the enhancement was also inhibited by antioxidants. Sulforaphane and auranofin, activators of Nrf2 that is a master regulator of anti-oxidative response, did not affect U46619-evoked edema but almost abolished TCDD-induced edema and potentiation by paraquat in both TCDD- and U46619-induced edema. Taken together, the results suggest that oxidative stress augments pre-cardiac edema caused by TCDD via activation of thromboxane receptor-mediated signaling in developing zebrafish. As paraquat and other oxidative stress inducers used also are environmental pollutants, interaction between dioxin-like compounds and exogenous source of oxidative stress should also be considered.     

松嫩草地羊草种子发育进程、休眠特性及与盐碱耐性关系的研究

羊草(Leymus chinensis)是禾本科赖草属根茎型优质禾草,不但营养价值高、适口性好,而且具有较强的耐旱、耐寒和耐盐碱性。羊草既是东北松嫩平原的优势种,也是干旱与半干旱地区建植人工草地的优良草种。近年来,随着畜牧业的发展及生态环境治理力度的加大,人工羊草草地不断建植,为此,人们对羊草种子的需求量与品质的要求也越来越高。本文通过研究羊草种子的发育进程、休眠特性及与盐碱耐性的关系,一方面明确羊草种子的最适宜收获时间,为农业生产上收获高品质羊草种子提供科学依据,另一方面挖掘羊草种子的发芽潜能、深入解析抗逆机理,为提高其利用率及抗逆新品种的选育提供理论基础。主要研究结果与结论如下:(1)通过对羊草种子发育动态的研究结果表明,羊草种子在发育过程中,随着成熟度的不断提高,种子的颜色由绿色变为浅绿色,再变成黄色,最后变为棕黑色。种子千粒重不断增加,在盛花后33 d达到最大值,之后趋于恒定。含水量与种子浸出液电导率则呈下降趋势,含水量在盛花后36 d达到最小值,而2个试验年份电导率值有所差异,分别在盛花后27和30 d达到最小值。标准发芽试验结果显示,羊草种子在盛花后39 d发芽率最高,此时种子的开始发芽时间、50%种子发芽时间、发芽势等指标均为最优。尽管加速老化试验的发芽指标与标准发芽试验略有差异,但是盛花后39 d的种子同样具有最强的抗老化能力。上述试验结果表明,盛花后39 d羊草种子活力最高,品质最佳,是种子最适宜的收获时间。 (2)不同成熟度的羊草种子对土壤埋深与盐碱胁迫具有不同的响应方式。羊草种子出苗与其后的幼苗生长能力随着种埋深度的增加而降低,1 cm是最适宜的播种深度,此时的出苗率最高、出苗时间最短,并且叶片与根系长度与生物量最大;另外不同成熟度的羊草种子表现出不同的出土成苗能力,盛花后39 d的羊草种子活力最大,其上述各项幼苗生长指标均为最优。种子的成熟度与盐、碱胁迫及其交互作用显著影响羊草种子的发芽率与发芽势,盛花后39 d的羊草种子在胁迫下具有最高的发芽率与发芽势,特别是在高浓度(400 mM)盐胁迫下,尤为明显。复萌试验结果显示,盛花后39 d的羊草种子在盐碱胁迫(特别是高盐环境)解除后同样具有最高的发芽率。上述结果表明,尽管不同成熟度的羊草种子均具有发芽能力,但是盛花后39 d的羊草种子出苗及抵御盐碱胁迫伤害的能力最强,这也进一步支撑了39 d是羊草种子适宜收获时间这一结论。另外羊草适宜浅播,1 cm是其最适宜的播种深度。 (3)通过人工手段处理可以明显打破羊草种子的休眠特性。研究结果表明,除了热水浸种处理外,其余方法如浓硫酸、冷层积、PEG,GA₃,KNO₃及清水浸种均能一定程度上提高羊草种子的发芽率,发芽速率、开始发芽时间及50%种子发芽时间。但是在生产实际中,既要考虑高效性也要考虑经济耗费,结合本试验的研究结果,我们推荐在生产中采用低温浸种20 d的方法来打破羊草种子的休眠,提高其发芽率。 (4)稃是抑制羊草种子萌发的重要因素,但同时也一定程度提高了种子的抗盐性。通过测定稃对羊草种子吸水、脱水、不同温度条件下的发芽响应以及不同持续时间盐胁迫对种子发芽的影响,结果发现稃可以显著提高羊草种子的吸水量,并同时减缓种子在干旱环境下的脱水速率,使种子不会过度脱水而死亡。稃、不同温度处理及两者交互作用显著降低羊草种子的发芽率与发芽速率,表明稃对羊草种子萌发具有一定的抑制作用。在不同持续时间的盐胁迫处理下,未萌发的带稃种子复萌率均高于去稃种子,特别是在长时间及高盐胁迫下尤为明显,表明稃对羊草种子耐盐性起着重要的调节作用,一旦雨水、融雪等条件降低了土壤盐浓度,带稃种子就可以继续萌发出土。 (5)20~30℃是羊草种子最适宜的发芽温度,高温、低温均显著降低种子的发芽率与发芽速率,并且此温度可一定程度上减缓盐胁迫与碱胁迫对种子发芽的抑制效应。随着盐、碱胁迫浓度的增加,羊草种子发芽率与发芽速率均呈下降趋势,且在碱胁迫下的下降幅度更大。在盐胁迫下,当盐浓度<200 mM时,低温是影响种子萌发的主要因素,随着盐浓度的不断增加,高温则更加剧了盐胁迫对种子萌发的抑制作用;而在碱胁迫下,即使碱浓度较低,高温与其交互作用也大大加剧了对种子发芽的抑制。盐胁迫下未萌发的羊草种复萌率随盐浓度增加而增加,而在碱胁迫下则随着碱浓度增加呈先上升后下降的趋势,高浓度碱胁迫使羊草种子失去活力而死亡,并且碱胁迫下种子的复萌率低于盐胁迫,25~35℃同样不利于种子的复萌。幼苗生长对温度与盐碱胁迫交互作用的响应方式与发芽阶段相似,20~30℃同样是最适宜温度;另外,盐碱胁迫均对羊草幼苗根生长的抑制作用更强。基于以上研究结果,我们建议在初夏(7月上旬),高降雨过后,温度与土壤条件适宜的情况下进行播种,以提高羊草种子的发芽率,更好的达到恢复退化草地的效果。 (6)在混合盐碱胁迫下,羊草种子的发芽率与发芽速率均随着盐浓度的增加不断下降,且碱性盐比例越大下降越明显。在250 mM盐浓度下,无碱性盐的A组处理发芽率为6.5%,而其余5组处理发芽率均为0。羊草幼苗生长阶段同样受盐浓度、pH及2者交互作用影响,并且根系对胁迫伤害更敏感,所受抑制作用更强。逐级回归分析结果表明,在种子萌发阶段,盐浓度是羊草种子在混合盐碱胁迫下能否萌发的决定性因素,而一旦胚根突破种皮进入幼苗生长阶段,pH就转变为主导因素。上述研究表明,混合盐碱胁迫对羊草种子萌发与早期幼苗生长阶段的抑制机理有所不同,其中高盐浓度与高pH的交互作用对种子萌发与幼苗生长的抑制效应最强。 (7)盐胁迫与碱胁迫均显著降低羊草幼苗的长度、鲜重与含水量,且碱胁迫抑制作用更强。2种胁迫均造成羊草幼苗Na⁺浓度与Na⁺/K⁺升高,并且K⁺浓度下降,但是在碱胁迫下,Na⁺ 浓度、Na⁺/K⁺上升幅度与K⁺下降幅度均大于盐胁迫。另外,在盐胁迫下,羊草幼苗大量积累Cl^-,有机酸含量变化不大;而在碱胁迫下,Cl^-,NO₃^-与H₂PO₄^-均呈下降趋势,而有机酸则大量积累,其中苹果酸、柠檬酸是主要的有机酸组分,可溶性糖是羊草幼苗在两种胁迫下共同的渗透调节物质。上述结果表明,碱胁迫由于具有高pH,对羊草早期幼苗生长的抑制作用更强,Cl^-与有机酸积累特征的差异表明羊草早期幼苗在盐胁迫与碱胁迫下具有不同的适应策略。