Measurement of Faecal Cortisol Metabolites in Cats and Dogs: A Non-invasive Method for Evaluating Adrenocortical Function

The aim of this comparative study was to gain more information about the metabolism and excretion of glucocorticoids in cats and dogs in order to establish non-invasive methods for evaluating stressful conditions. Therefore, in a first experiment, [¹⁴C]cortisol was administered intravenously to 8 animals (two of each sex and species). Over a period of 6 days, faeces and urine were collected immediately after spontaneous defecation and urination. Marked species differences were found, as cats mainly excreted cortisol in the faeces (82%±4% of the total recovered radioactivity), whereas in dogs only a small portion was found there (23%±4%). The highest urinary radioactivity was observed after 9±3 h in cats and 3±1 h in dogs. Peak concentrations in the faeces occurred after 22±6 h in cats and after 24±4 h in dogs. Most of the radioactivity was not extractable with diethyl ether, indicating that the metabolites excreted in urine and faeces were mainly of the conjugated or polar unconjugated types. This was confirmed by RP-HPLC, which also revealed marked differences between cats and dogs concerning the metabolites formed. In addition, the immunoreactivity of the metabolites was tested in cortisol, corticosterone and 11-oxoaetiocholanolone EIAs. The latter, measuring 11,17-dioxoandrostanes (11,17-DOA) detected the highest quantities of immunoreactive metabolites in cats, but not in dogs. In a second experiment, the adrenal cortex of both species was stimulated by ACTH and, three weeks later, suppressed by dexamethasone. In this study, only faeces were collected over a period of 7 days. In both species, inter-animal variability in the basal and maximal/minimal faecal cortisol metabolite concentrations and the time course was observed. The 11-oxoaetiocholanolone EIA in cats and the cortisol EIA in dogs proved best suited for monitoring changes in adrenocortical activity. ACTH injections resulted in an increase above baseline values of 355% (median) in 11,17-DOA concentrations in cats and of 702% in the concentrations of cortisol equivalents in dogs by about 25 h and 22 h (median) after injection, respectively. Minimal concentrations after dexamethasone administration were about 17% in cats and 31% in dogs (in relation to baseline values) and were reached in 66 h and 72 h, respectively. It was concluded that measuring cortisol metabolites in faeces should be a useful non-invasive tool for monitoring stress in carnivores.     

盐碱胁迫下黑麦草生长及离子微区分布特征

盐碱生境通过离子毒害和渗透胁迫等抑制植物的生长发育。离子微区分布调控主要通过调整盐分在不同部位的分布来减轻Na⁺等高浓度离子对重要组织器官的毒害作用,是盐碱胁迫下植物生存的重要策略之一。为探究黑麦草在盐碱胁迫下的离子微区分布特征,本研究通过水培法,采用不同盐碱浓度(0、50、100、200 mmol·L^-1)培养液处理黑麦草植株,考察其生长、离子吸收、运输选择性和微区分布变化,揭示黑麦草对盐碱胁迫的适应机制。结果表明:1) 盐碱胁迫下,植株的相对含水率、叶绿素含量逐渐降低,电解质外渗率显著升高;当盐碱浓度大于100 mmol·L^-1时,根长、株高和生物量均显著降低,即黑麦草的耐盐阈值是100~200 mmol·L^-1;2) 盐碱浓度为50 mmol·L^-1时,植株根中K⁺/Na⁺较空白升高了48.3%,Ca²⁺/Na⁺升高了54.1%,说明黑麦草根部可通过增强K⁺、Ca²⁺的吸收来维持细胞渗透压并缓解高浓度Na⁺的毒性;3) 在50~200 mmol·L^-1的盐碱浓度下,黑麦草叶片中可溶性组分及细胞壁中Na⁺的相对含量较对照显著增加,但细胞器内Na⁺的相对含量降低,说明叶部将Na⁺富集在液泡和细胞壁中,以减少Na⁺对细胞器的毒害作用;4) 叶片微观结构的SEM电镜图片显示,盐碱胁迫下叶片表皮增厚、导管数量减少和孔径缩小。综上所述,盐碱胁迫下黑麦草不同器官的适应机理不同,根部主要通过强化K⁺、Ca²⁺的吸收,降低对Na⁺的吸收,并将Na⁺区隔化在液泡中以降低离子毒害和维持渗透调节,而叶片主要通过将Na⁺区隔在液泡及细胞壁中,以及微观结构的变化来保护细胞器免受离子毒害。本研究可为黑麦草的耐盐碱适应机制及其在盐碱土上的种植提供理论依据。     

盐渍地区草坪草品种及其混播组合筛选的研究

对 11种草坪草室内发芽试验表明 :盐浓度 0 %～ 1.6 %NaCl胁迫下相对发芽率均随盐浓度升高而下降 ,呈线性回归关系。在北方盐渍地上 0～ 2 0cm土层全盐含量不超过 1.0 %时 ,混播优于单播。表现较好的混播是多年生黑麦草 +碱茅 +草地早熟禾或匍匐翦股颖。其中多年生黑麦草发芽快、幼苗生长迅速 ,很快覆盖地面 ,有利于其他种子的萌发 ,称为保护种。碱茅幼苗期及成株后耐盐性均较强 ,但苗期生长慢 ,适宜与多年生黑麦草伴生。草地早熟禾和翦股颖品种中耐盐性较强的可作为主要草种     

盐胁迫下匍匐翦股颖抗氧化酶活性及基因表达机制研究

为研究匍匐翦股颖在盐胁迫下抗氧化酶活性与基因表达的影响机制,模拟盐胁迫条件,用0 mol/L(CK),0.2 mol/L的NaCl溶液对两种不同耐盐性的匍匐翦股颖Penncross和SeasideⅡ进行处理,测定了超氧化物歧化酶(SOD),过氧化氢酶(CAT),抗坏血酸过氧化物酶(APX)和过氧化物酶(POD)活性;用同源克隆的方法克隆了匍匐翦股颖CytSOD,FeSOD,CAT,APX,POD的部分片段,并设计了5个基因的qRT-PC引物,对5个抗氧化酶基因表达量进行了荧光实时定量测定,分析了两种不同耐盐性的匍匐翦股颖在盐胁迫条件下,抗氧化酶活性与基因表达的变化规律。结果表明,盐胁迫对两种不同耐盐性的匍匐翦股颖抗氧化酶活性和基因表达的影响不同,在盐胁迫初期,盐敏感的Penncross SOD活性比耐盐的SeasideⅡ高,盐处理中、后期Penncross SOD活性快速下降,SeasideⅡSOD活性下降慢而且能保持相对稳定,在这一阶段SeasideⅡSOD活性显著高于Penncross;CAT,APX和POD活性随着盐处理的延长,酶活性增加,Penncross CAT和APX活性低于SeasideⅡ,POD活性两者差异不大。盐胁迫下,Penncross CytCu/ZnSOD,CAT、APX 和POD表达量上调,而FeSOD下调;SeasideⅡ5种抗氧化酶基因表达量上调,CytCu/ZnSOD,FeSOD,CAT和APX的表达量显著高于Penncross,而POD表达量差异不大。两种匍匐翦股颖CytCu/ZnSOD、CAT、APX和POD表达量变化与酶的活性变化较一致,而盐敏感的FeSOD的表达量变化与SOD活性变化不一致。     

淹水对紫花苜蓿南北方品种抗氧化酶和无氧呼吸酶的影响

以紫花苜蓿南方育成品种渝苜1号和北方地方品种新疆紫泥泉为材料,在幼苗长出4片真叶时进行淹水处理,在淹水的第10天测定植株的生物量,并分别在淹水的第0,2,4,6,8,10天测定叶片中的丙二醛(MDA)含量以及叶片抗氧化酶和根系无氧呼吸酶的活性,以探明两个品种对淹水胁迫的耐受性差异及其生理响应机制。结果表明,淹水胁迫使紫花苜蓿的生物量减少,但与新疆紫泥泉相比,渝苜1号生物量的降幅小。淹水时渝苜1号叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)等抗氧化酶活性比新疆紫泥泉的高,而叶片MDA含量比新疆紫泥泉的低;根系无氧呼吸酶活性在淹水胁迫中均增强,但渝苜1号的乙醇脱氢酶(ADH)和丙酮酸脱羧酶(PDC)活性增加明显,乙醇发酵更强;而新疆紫泥泉的乳酸脱氢酶(LDH)的活性增加更为突出,乳酸发酵强。淹水影响紫花苜蓿正常生长,但南方品种渝苜1号比北方品种新疆紫泥泉对淹水胁迫更具耐受性,因为前者具有比较高的抗氧化酶活性和比较强的乙醇发酵途径,有利于增强植株抗淹水胁迫能力。     

模拟干旱和盐胁迫下沙冬青种子硬实特性和抗逆性的研究

将1~5 d内每天吸胀的沙冬青(Ammopiptanthus mongolicus)种子视为不同硬实程度的种子T₁,T₂,T₃,T₄和T₅,与第5 d仍未吸胀的硬实种子(记作T_r)作为试材对其硬实特性和抗逆性进行研究,以期为沙冬青种子栽培驯化提供理论依据。结果表明:随着硬实程度的提高,种子的发芽势、发芽指数、活力指数呈上升趋势;电导率和丙二醛(MDA)含量逐渐下降;可溶性蛋白含量逐渐上升。不同处理对种子的硬实破除效果表明,80℃热水浸泡种子可达到最佳破除效果。通过测定模拟干旱胁迫和盐胁迫下不同硬实程度种子的超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性和脯氨酸(Pro)含量,表明在相同逆境条件下,随硬实程度的加深,除极个别外,各项指标值均有不同程度的升高,且一定胁迫强度下硬实种子萌发幼苗的各酶活性和脯氨酸含量均极显著高于其他萌发幼苗(P<0.01),因此,硬实种子比非硬实种子有较强的保护酶调节系统。硬实程度越高,耐旱和耐盐能力也随之升高。     

发酵豆粕对仔猪早期断奶应激及增重效果的影响

仔猪早期断奶是提高母猪繁殖利用率的重要措施,但早期断奶应激容易导致仔猪断奶后食欲下降,消化机能紊乱,腹泻等“仔猪早期断奶综合症”.发酵豆粕因富含植物小肽,易于动物的消化吸收等多种优点,适用于幼龄、病弱、应激等特殊生理状况下的仔猪.本试验选用市场上常用的一种同类产品与一种新开发的发酵豆粕在断奶仔猪上进行饲喂对比.结果表明:这种新开发的产品对缓解仔猪早期断奶应激引起的胃肠机能紊乱明显优于对照产品,其稀便发生率显著低于对照组(P<0.01);平均日增重、料肉比高于对照组,但差异不显著(P>0.05).     

Physiological but not morphological adjustments along latitudinal gradients in a human commensal species,the Eurasian tree sparrow

Human commensal species take advantage of anthropogenic conditions that are less likely to be challenged by the selective pressures of natural environments. Their morphological and physiological phenotypes can therefore dissociate from habitat characteristics. Understanding how these species adjust their morphological and physiological traits across latitudinal gradients is fundamental to uncovering the eco-physiological strategies underlying coping mechanisms. Here, we studied morphological traits in breeding Eurasian tree sparrows (ETSs; Passer montanus) among low-latitude (Yunnan and Hunan) and middle-latitude (Hebei) localities in China. We then compared body mass; lengths of bill, tarsometatarsus, wing, total body, and tail feather; and baseline and capture stress-induced levels of plasma corticosterone (CORT) and the metabolites including glucose (Glu), total triglyceride (TG), free fatty acid (FFA), total protein, and uric acid (UA). None of the measured morphological parameters varied with latitude except in the Hunan population, which demonstrated longer bills than those in other populations. Stress-induced CORT levels significantly exceeded baseline levels and decreased with increasing latitude, but total integrated CORT levels did not vary with latitude. Capture stress-induced significantly increased Glu levels and decreased TG levels, independent of site. However, the Hunan population had significantly higher baseline CORT, baseline and stress-induced FFA levels, but lower UA levels, which differed from other populations. Our results suggest that rather than morphological adjustments, physiological adjustments are mainly involved in coping mechanisms for middle-latitude adaptation in ETSs. It is worth investigating whether other avian species also exhibit such dissociation from external morphological designs while depending on physiological adjustments.     

磷素营养对星星草幼苗抗碱性的影响

通过以不同水平磷素营养培养星星草幼苗，研究磷素营养对星星草幼苗抗碱性的影响，试验结果表明：在２．０％Ｎａ２ＣＯ２胁迫下星星草苗磷素营养的最适浓度为２．８ｍｍｏｌ·Ｌ＾－１ＫＨ２ＰＯ４，此条件下仅对株高，干重和鲜重的增长有促进作用，而且受害程度最小，对Ｎａ２ＣＯ３胁迫的缓解作用最强，在一定程度上可提高星星草幼苗对Ｎａ２ＣＯ３胁迫的抗性。     

雏鸡冷应激前后血清中差异蛋白质组学研究

本试验旨在比较分析雏鸡冷应激前后血清中蛋白质的表达差异,并对重点差异蛋白质进行鉴定。将30只雏鸡随机分为3组,分别为冷应激组、冷适应组和常温对照组,收集各组雏鸡的血液制备血清后进行双向凝胶电泳(2-DE),以获得血清蛋白质表达的2-DE图谱,对2-DE图谱进行差异分析(利用PDQuest 8.0软件),随后采用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术对差异表达蛋白质进行鉴定,并利用蛋白质印迹(Western blot)方法进行验证。结果显示:通过2-DE对常温对照组、冷应激组和冷适应组雏鸡血清进行分析,得到了比较完整的差异蛋白质数据,共找到差异蛋白质点23个。采用MALDI-TOF-MS技术分析其中几个重复性好且较为明显的蛋白质点,成功鉴定出4个差异蛋白质,其中2个为果糖二磷酸醛缩酶C(ALDOC),是参与葡萄糖、能量代谢通路供能的相关蛋白;随后,对差异蛋白质ALDOC进行Western blot验证,所得结果与2-DE的结果相一致。结果表明,雏鸡冷应激前后血清中蛋白质的表达具有明显的差异,这些蛋白质的表达差异可能与冷应激有关。