Detection of HSP mRNA Transcription in Transport Stressed Pigs by Fluorescence Quantitative RT-PCR

The RNA transcripted in vitro was used as the standard quantitative template to make the standard curve and establish the fluorescence quantitative RT-PCR （FQ-PCR） method. By means of FQ-PCR, the transcription changes of HSP70 and HSP90 mRNA in the livers and hearts of transport stressed pigs were studied. The level of HSP70 mRNA transcription increased continuously from the beginning of transportation. The inductions of HSP70 mRNA transcription in the livers and hearts of 10 h transport stressed pigs were 2.5 and 4.1 times higher than that of the un-transport stressed pigs （P〈0.01）. However, the transcription levels of HSP90 mRNA in the livers and hearts decreased with the transport stress.

Effects of transport stress on immune response, physiological state, and WSSV concentration in the red swamp crayfish Procambarus clarkii

Transport is an essential part of the aquaculture and research of the main freshwater aquaculture crayfish Procambarus clarkii in China.However,transport is often accompanied by a low survival rate.Assessing the physiological state of P.clarkii before and after transport may discover the cause of this high mortality rate.In this study,ice-cold and exposed-to-air transport methods were compared using an array of parameters,including relative expression level of heat shock protein 70(HSP70),content of serum glucose and cortisol,immune parameters(enzyme and immune-related genes),and white spot syndrome virus(WSSV)concentration were investigated to understand the physiological state of P.clarkii before and after transport,as well as the cause of dying crayfish on days 5 and 7 after transport stress.Histological sections of hepatopancreas,gills,and intestines reflected pathological changes.The survival rate of crayfish with ice-cold transport was significantly higher than that with exposed-to-air transport,and mortality peaked at 3-9 days after transport stress.A prolonged response to oxidative stress and short-term immunosuppression was present after transport,and the trend of the WSSV concentration in the hepatopancreas was similar to the mortality rate of P.clarkii.The contents of serum glucose and cortisol,antioxidant enzymes and immune-related indexes,and the concentration of WSSV in hepatopancreas of dying crayfish were significantly higher than those of vibrant crayfish on the 5th and 7th days after transport.The hepatopancreas,intestines,and gills of dying crayfish had varying degrees of damage,and the hepatopancreas and intestines were severely damaged.The results suggested that the death of P.clarkii after transport stress is caused by oxidative stress,the imbalance of reactive oxygen species regulation,and decreased WSSV resistance,which eventually led to irreversible tissue damage.The increase of WSSV in the body of crayfish might be the direct cause of crayfish death.

EFFECTS OF TRANSPORT STRESS ON THE INTESTINES INVOLVING NEURONAL NITRIC OXIDE SYNTHASE

Transport stress is commonly suffered by animals with gastrointestinal dysfunction a common symptom.Currently,the mechanisms of transport stress-induced intestine impairment are largely unknown.The aim of this study was to investigate the effects of transport stress on the expression of neuronal nitric oxide synthase(nNOS)and the distribution of nNOS-positive neurons of the intestines in rats and to explore the neuroendocrine mechanism of transport stress.In this study,Sprague Dawley rats(n=6)were subjected on a constant temperature shaker for 1(S1d)or 3 d(S3d).Rats exhibited increased serum glucose and diminished total number of leukocytes,in which lymphocytes level was also decreased in the S1d group(P<0.05).Also,normal intestinal morphology was disrupted in the S1d rats,and the thickness of muscle layers was decreased in duodenum,jejunum and colon of S3d rats.In addition,it was found that nNOS expression,as well as the number of nNOSpositive neurons in the myenteric plexus were downregulated in duodenum,jejunum and colon of S3d rats compared with that of unstressed rats(P<0.05).These data reveals that transport stress induced intestinal damage and uncovers potential action mechanisms that nNOS-positive neurons and nNOS expression might be involved in modulating this process.

Thermoelastic stresses in SiC single crystals grown by the physical yapor transport method

A finite element-based thermoelastic anisotropic stress model for hexagonal silicon carbide polytype is developed for the calculation of thermal stresses in SiC crystals grown by the physical vapor transport method. The composite structure of the growing SiC crystal and graphite lid is considered in the model. The thermal expansion match between the crucible lid and SiC crystal is studied for the first time. The influence of thermal stress on the dislocation density and crystal quality is discussed.

Glutamate Transporter 1-mediated Antidepressant-like Effect in a Rat Model of Chronic Unpredictable Stress

In recent years, more attention has been paid to the role of the glutamate transporter 1 （GLT-1, EAAT2） in major depressive disorder （MDD）. However, experimental data on brain GLT-1 levels are, to some extent, inconsistent in human postmortem and animal studies, These discrepancies imply that the role of GLT-1 in the pathophysiology of MDD and the action of antidepressants remain obscure. This work was designed to study the impact of chronic unpredictable stress （CUS） for 2 ses- sions per day for 35 days and four weeks of fluoxetine （FLX） on depressive-like behaviors in rats, as well as the concomitant expression of the GLT-1 protein in the hippocampus. Behavioral changes were assessed by the sucrose preference and open field tests. GLT-1 levels were detected by immunohisto- chemistry and Western blot analysis. Our study demonstrated that the animals exposed to CUS showed depressive-like behaviors and exhibited a significant decrease in GLT-1 expression in the hippocampus. Chronic FLX treatment reversed the behavioral deficits and the CUS-induced decrease in GLT-1 levels. Taken together, our results support the reduction of GLT-1 in human postmortem studies in MDD and suggest that GLT-1 may be involved in the antidepressant activity of FLX. Our studies further support the notion that GLT-1 is an attractive candidate molecule associated with the fundamental processes of MDD and may be a potential, and novel pharmacological target for the treatment of MDD.

Effects of Transportation Stress during the Hot-Dry Season on Some Haematological and Physiological Parameters in Moroccan Dromedary Camels （Camelus dromedarius）

The purpose of this study was to determine the effects of road transportation under heat conditions on some haematological [Ht （haematocrit）, blood cells count and EOF （erythrocytes osmotic fragility）] and physiological [Tr （rectal temperature）, HR （heart） and RR （respiratory rates）, and circulating levels of Cor （cortisol）, Glu （glucose） and minerals] parameters in Moroccan dromedary camels. The animals were subjected to road transportation stressor for 2 h by truck during the hot-dry season. Blood samples were collected before loading and transport, and at the end of transport. Transportation induced a significant increase （P 〈 0.05） of erythrocytes count, Ht, EOF, Tr, HR and RR by comparison to values observed before transportation. The same stress conditions induced a significant increase （P 〈 0.05） of plasma Cor （ng/mL） and blood Glu （mM） （220 ± 30 vs. 137 ± 20, 9.7 ± 1.2 vs. 6.4 ± 1. 1 respectively） and a significant decrease （P 〈 0.05） of plasma magnesium （mM） （0.5 ± 0.1 vs. 0.9 ± 0.1） comparatively to pre-transportation values. These results indicate that road transportation associated to heat may be considered as a potent stressor which is able to induce several cellular alterations in camels. Further studies of an eventual protective role of vitamin C against haemolysis induced by transportation stress in camel are needed.

Evaluation of Stress Corrosion Cracking Damage to an API 5L X52 Pipeline Transporting Ammonia: A Case Study

The high number of leak events that took place in recent years at a 25.4 cm (10”)&Oslash;pipeline transporting anhydrous liquid ammonia, located in the Southeast of Mexico, was the main reason to carry out a number of field studies and laboratory tests that helped establish not only the failure causes but also mitigation and control solutions. The performed activities included direct evaluation at failure sites, total repair programs, metallographic studies and pipeline flexibility analyses. The obtained results were useful to conclude that the failures obeyed a cracking mechanism by Stress Corrosion Cracking (SCC) which was caused by the combined effect of different factors: high stress resistance, high hardness of the base metal with a microstructure prone to brittleness and residual strains originated during the pipeline construction. From the operative, logistic and financial standpoints, it is not feasible to release the stress of approximately 22 km of pipeline. Therefore, the only viable solution is to install a new pipeline with suitable fabrication, construction and installation specifications aimed at preventing the SCC phenomenon.

维生素C对禁食黄颡鱼抗氧化能力和肌肉品质的影响

【目的】研究添加不同浓度维生素C对禁食暂养黄颡鱼(Pelteobagrus fulvidraco)抗氧化能力和肌肉品质的影响,为提升鲜活黄颡鱼耐运性和肌肉品质提供理论依据。【方法】随机将黄颡鱼分为4组:T0-0(0 mg/L V_(C),暂养0 d)、T0-5(0 mg/L V_(C),暂养5 d)、T10-5(10 mg/L V_(C),暂养5 d)、T30-5(30 mg/L V_(C),暂养5 d),每组40尾鱼,检测血清、肌肉的抗氧化物质以及肌肉品质指标。【结果】T5-0组黄颡鱼血清中GSH-PX、CAT、GSH活性以及MDA浓度,肌肉中CAT、GSH活性和MDA浓度显著低于T1-0组(P<0.05);其纤维结构变得紧密,pH下降,白度增加,持水力增加,肌肉品质指标显著升高(P<0.05)。与T0-5及T30-5组相比,T10-5组血清中SOD、CAT、GSH、GSH-PX活性显著较高(P<0.05),MDA浓度较低;肌肉中SOD、GSH活性与血清中结果变化相同;持水力进一步增加。在质构方面,T10-5组剪切力、硬度、弹性、咀嚼性、黏性均高于T0-5组及T30-5组。【结论】添加10 mg/L的V_(C)能进一步显著提高黄颡鱼的抗氧化能力和肌肉品质(P<0.05),但30 mg/L V_(C)添加不利于黄颡鱼肌肉品质的提升,甚至对其产生负面影响。

西瓜蔗糖转运蛋白SUT家族的鉴定及其在果实发育和逆境响应中的表达分析

蔗糖转运蛋白(SUTs)作为蔗糖主动转运的主要载体,在分配同化物从源到库组织的转运中起着关键作用。尽管SUTs的特性及其生物学功能已在高等植物中得到了深入研究,但该基因家族尚未在西瓜中进行鉴定。本研究在西瓜基因组中共鉴定了4个ClSUT基因,并对其基因和蛋白结构、保守基序、亚细胞定位、染色体分布、进化关系、启动子元件等进行了全面分析。基因结构分析表明,亚族Ⅰ的ClSUT3和ClSUT4基因均只含有一个内含子。所有ClSUT均被预测定位在质膜中,它们均含有一个保守的MFS-2结构域,并且含有与之对应的5个保守基序。系统发育分析发现拟南芥、水稻和西瓜中的SUT分为5个亚族,西瓜ClSUT与拟南芥SUT的亲缘关系更近,被聚类在亚族Ⅰ、亚族Ⅱ和亚族Ⅴ。还在西瓜ClSUT基因的启动子中鉴定到一些与激素和逆境响应相关的作用元件。此外,转录组数据表明,西瓜ClSUT基因的表达对渗透和盐胁迫呈现多样性,很可能参与西瓜对这些逆境的响应。

樟树对铜胁迫的生长和生理响应及铜富集转运特性分析

以前期在铜污染矿区修复中筛选出的耐铜樟树(Cinnamomum camphora(L.)Presl)为材料,通过盆栽模拟实验,以不添加铜为对照(CK),设置50、150、300、600、900、1200 mg/kg 6个处理浓度,胁迫处理60 d后,测定樟树的生物量积累、叶绿素含量、生理生化指标及樟树对铜离子的富集和转移量,分析不同浓度铜处理对耐铜樟树生长、生理生化的影响,以及铜在不同组织的富集转运规律。结果显示,樟树对铜胁迫表现出低浓度(150 mg/kg)促进生长,高浓度抑制生长的趋势。超氧化物歧化酶(Superoxide dismutase,SOD)、过氧化物酶(Peroxidase,POD)、过氧化氢酶(Catalase,CAT)、过氧化氢(Hydrogen peroxide,H_(2)O_(2))、游离脯氨酸和可溶性糖含量等生理指标在900 mg/kg处理时没有显著升高;谷胱甘肽还原酶(Glutathione reductase,GR)随着处理浓度的升高逐渐降低;丙二醛(Malondialdehyde,MDA)、谷胱甘肽(Glutathione,GSH)含量则随着处理浓度的升高逐渐升高。各器官铜含量的大小顺序为根>叶>茎,樟树主要将铜富集在根系,往地上茎和叶的转移率很低,极大地降低了对茎、叶的铜毒害作用,且随着处理浓度的增加,铜由根系向叶片的转移率IF(Leaf/Root)由对照处理的0.078逐降至1200 mg/kg处理时的0.007;铜由根系向茎的转移率IF(Stem/Root)由对照处理的0.06逐降至1200 mg/kg处理时的0.005。综合其生长、生理及铜的富集转运特性,樟树对铜具有较好的耐受性,耐受浓度可达900 mg/kg,是可用于铜污染土壤种植的理想材料。

小麦钼营养研究进展

钼是小麦生长所必需的一种微量元素,对小麦的生长发育有重要作用,影响小麦的生理代谢、产量和品质等。本文综述了目前我国土壤中钼含量及其有效性,施钼对小麦生理代谢、钼的吸收分配、吸收效率及其基因表达的影响,以及施钼对小麦生长发育、籽粒产量和品质及其抗逆性影响等的研究现状,并对今后开展小麦钼营养研究的重点方向进行了展望。

道路运输对羔羊体重、血液生化及临床指标的影响

试验旨在研究道路运输对羔羊体重、血液生化指标和临床指标的影响,为生产中有效缓解羔羊运输应激提供理论依据。选取平均体重(21.52±1.22)kg健康4月龄羔羊10只进行8 h的长途运输,分别在运输前后进行称重、静脉采血,并测定体温、呼吸频率、脉搏等临床指标。结果表明:①道路运输使羔羊体温、呼吸和脉搏频率极显著升高(P<0.01),体重降低了15.31%(P<0.05)。②道路运输极显著升高了羔羊血清中促肾上腺皮质激素(ACTH)和乳酸脱氢酶(LDH)水平(P<0.01),显著升高了皮质醇(COR)和肌酸激酶(CK)水平(P<0.05),血清中谷丙转氨酶(ALT)水平无显著变化(P>0.05)。③道路运输显著降低了羔羊血清中免疫球蛋白A(IgA)、免疫球蛋白G(IgG)水平(P<0.05),显著升高了白介素-6(IL-6)水平(P<0.05),血清中免疫球蛋白M(IgM)、干扰素-γ(IFN-γ)、肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)水平与运输前差异不显著(P>0.05)。④道路运输显著降低了羔羊血清中葡萄糖(GLU)、总蛋白(TP)、白蛋白(ALB)水平(P<0.05)。⑤道路运输显著降低了羔羊血清中红细胞总数(RBC)、红细胞压积(HCT)、血红蛋白含量(MCH)(P<0.05),红细胞平均体积(MCV)极显著降低(P<0.01),白细胞总数(WBC)显著升高(P<0.05)。综上所述,羔羊经过8 h道路运输后发生了明显的应激反应,导致羔羊体重下降,机体代谢增强,炎症反应增强,免疫系统受到了一定程度损伤。

鱼类运输应激反应及其对存活率影响研究进展

【目的】探讨促进活鱼类运输产业中关注的提高存活率问题的解决方案。【方法】在理解鱼类应激生理反应的基础上,对适用于运输中的应激指标进行分类,分析其应用局限性和发展趋势;保活运输过程中的应激源可分为温度、拥挤、溶氧、氨氮、盐离子、振动和噪声等,综述不同应激源对应激指标和存活率的影响。【结果与结论】在保活运输过程中,应当系统、全面地分析多项应激指标,总结其趋势和相互关联,以正确判定应激反应的强度;同时考虑鱼种、季节、鱼体大小、运输时间等因素的影响,进而制定适宜的保活方式,提高鱼的存活率。

保活运输应激对水产动物肌肉品质影响的研究进展

【目的】揭示应激对水产动物肌肉品质的影响,以期为水产动物保活运输提供参考。【方法】将保活运输过程中的应激源分为温度、低氧、拥挤、振动、氨氮和空气暴露等,综述各种应激源对不同水产动物肌肉品质的影响;在此基础上,概括加盐、暂养、加抗氧化剂、休眠和冷驯化等减缓应激措施对水产动物肌肉品质的影响,总结组学技术在揭示应激影响水产动物肌肉品质机制的应用。【结果与结论】应激状态下,水产动物的生命活动中,如基因表达、代谢和免疫等都会进行适应性调控,使蛋白质、代谢物组成和细胞形态等发生改变,并最终影响肌肉品质。消除应激可提升水产动物肌肉品质,关于其对提升肌肉品质的机理方面研究是未来的研究方向。

水稻对恶苗病菌侵染响应的转录组分析

水稻是主要的粮食作物,而恶苗病的发生对水稻生产构成了巨大威胁,造成一些品种严重减产。转录组测序技术已逐渐应用到水稻恶苗病的品种抗性研究中,使用高抗和高感水稻品种,经接种、取样和测序分析,明确水稻恶苗病在转录水平上的抗性机制。利用Illumina NovaSeQ6000平台构建了12个真核普通转录组文库并测序,以|log 2(FoldChange)|≥1及p≤0.05为标准,筛选出抗病品种比较组合(KDI vSKDCK)1482个差异基因,感病品种比较组合(LJI vSLJCK)2293个差异基因。通过共表达韦恩图展示,抗病比较组合(KDI vSKDCK)差异基因和感病比较组合(LJI vSLJCK)差异基因共表达328个,其中抗病比较组合(KDI vSKDCK)差异基因中唯一表达1154个,感病比较组合(LJI vSLJCK)差异基因中唯一表达1965个。进一步通过共表达韦恩图展示,抗病比较组合(KDI vSKDCK)上调表达差异基因和感病比较组合(LJI vSLJCK)上调表达差异基因共表达31个,抗病比较组合(KDI vSKDCK)上调表达差异基因中唯一表达272个,感病比较组合(LJI vSLJCK)上调表达差异基因中唯一表达825个。将抗病比较组合(KDI vSKDCK)上调表达差异基因中唯一表达的272个基因进行GO功能富集分析后,得到264个GO Term,其中,显著富集GO Term有1个生物过程(BP)和6个分子功能(MF),均与病原菌感染响应的氧化应激反应(GO:0006979)、过氧化物酶活性(GO:0004601)、氧化还原酶活性和作用于作为受体的过氧化物(GO:0016684)、抗氧化活性(GO:0016209)有关,显示抗病品种水稻芽在恶苗病菌侵染时参与氧化应激反应生物过程基因显著上调表达,ATP酶活性显著增加,促进物质跨膜转运,有效提高了抗病性。

非饱和土的有效应力探讨

由于非饱和土强度季节性变化,缺失统一的非饱和土有效应力公式而无法准确预警预防,导致非饱和土堆土、边坡出现滑坡、塌陷等诸多问题,基于Terzaghi的有效应力理论和土力学三相介质原理,推导得到非饱和土土颗粒之间的相互作用力,建立了非饱和土的有效应力公式.在此基础上,分别考虑应变功与能量守恒原理、非饱和土气水运移和受力平衡等不同情形,进一步论证了该非饱和土有效应力公式的科学性,同时利用不同的非饱和土剪切强度试验数据对这一有效应力公式进行计算验证,证明了其正确性和有效性.研究结果表明,所建立的非饱和土有效应力公式可适用于饱和土和非饱和土,并便于相关工程的设计和灾变防治应用.

肉鸡运输应激发生机制与防控策略

运输是肉鸡生产闭环中不可避免的过程,但由于运输过程中各种环境因素和外界刺激等的综合作用,可能会对肉鸡造成多种应激反应,导致鸡只自身免疫、繁殖机能和生产性能等出现不同程度下降,进而严重影响肉鸡生产企业的经济效益。文章列举了常见运输应激源的产生及其对肉鸡的影响,包括不合理的抓取装箱方式、禁食、急性冷热刺激、运输密度等;总结了运输应激的作用机理,包括内分泌轴应激应答系统以及氧化应激作用通路,并从提高抗应激能力和减少应激源两个角度提出内外结合预防运输应激的潜在有效手段,提出将运输应激的研究同精准畜牧技术、人工智能、医学等方向结合的展望,旨在为肉鸡运输相关的研究和生产提供参考。

辣椒CaSULTR基因家族的鉴定、表达与克隆

【目的】作为植物所需的四大营养元素之一,硫在植物生长发育、抵御生物和非生物胁迫中扮演着关键角色。硫酸盐转运蛋白(Sulfate transporter,SULTR)作为植物对硫酸盐吸收和运输的主要调节者,在这一过程中发挥着不可替代的作用。了解辣椒CaSULTR的基本特性及表达模式,为推进辣椒SULTR的研究提供理论支持。【方法】以辣椒基因组为基础,在全基因组范围内鉴定辣椒CaSULTR基因家族的成员,并对其理化性质、保守基序、基因结构等进行分析。基于已公布的转录组数据及荧光定量PCR技术,分析不同激素胁迫条件下辣椒CaSULTR基因的表达模式,并对差异表达基因进行克隆。【结果】共鉴定到13个辣椒CaSULTR基因,分别命名为CaSULTR1;1~1;3、CaSULTR2;1~2;2、CaSULTR3;1~3;7、CaSULTR4;1。这些基因分布在辣椒的7条染色体上,氨基酸长度为641~947 aa,分子量69.65~104.36 kD,等电点在8.42~9.62,均为疏水性蛋白。CaSULTR被分为4个亚组(Ⅰ~Ⅳ),其中Ⅲ亚组包含最多成员,同一亚组不同成员间的理化性质差异较小且Motif和基因结构相对保守,除CaSULTR2;1和CaSULTR2;2外的成员均具有Motif 5。组织表达分析显示,CaSULTR可能参与叶片、果实、花等器官的发育过程,其中CaSULTR4;1在叶、根、茎中均有表达。激素胁迫下的辣椒表达谱与荧光定量PCR结果显示,CaSULTR1;1和CaSULTR3;4受激素诱导后显著上调表达。【结论】明确了CaSULTR在激素胁迫下的表达模式,推测CaSULTR1;1和CaSULTR3;4可能参与外源激素调节硫代谢的过程。

玉米转录因子ZmNAC59调控植物抗盐性

【目的】NAC转录因子家族是植物中研究较多的一类转录因子,在调控植物生长发育和响应非生物胁迫过程中起重要作用。玉米是三大粮食作物之一,其生长过程常会面临各种逆境胁迫,盐害被认为是限制作物生长和生产的主要环境因素之一。因此鉴定玉米抗盐基因,解析其抗盐机制对培育玉米抗逆品种具有重要意义。【方法】研究克隆了玉米转录因子ZmNAC59,并用生物信息学手段分析其保守结构域和系统进化关系,用实时荧光定量PCR方法分析该基因在NaCl和MeJA处理下的表达模式,并用稳定转基因体系将该基因异源表达拟南芥观察表型,同时还用病毒诱导沉默技术将该基因在玉米中沉默后进行盐处理表型观察,并进行酶活性检测。【结果】ZmNAC59能够被NaCl和MeJA诱导上调。病毒诱导沉默ZmNAC59后进行盐胁迫,沉默株系对盐胁迫更敏感,ROS积累更多;而ZmNAC59过表达拟南芥后,过表达株系在盐胁迫处理下存活率更高,活性氧积累更少,Na^(+)/K^(+)比率低,表明ZmNAC59作为盐胁迫中的正调控因子可以通过调节离子流动提高植物抗盐性。实时荧光定量PCR结果表明拟南芥过表达株系中Na^(+)、K^(+)转运相关的基因显著上调;在玉米原生质体中瞬时过表达ZmNAC59后进行盐处理发现ZmSOS1、ZmNHX1和ZmNHX7的表达显著上调,Dual-LUC实验表明ZmSOS1是Zm-NAC59的靶基因。【结论】研究表明ZmNAC59通过激活ZmSOS1启动子的活性调控其表达而促进盐胁迫下的离子转运,从而提高植物抗盐性,为玉米抗逆基因资源的筛选和品种培育提供了科学理论依据。

畜禽运输应激及其缓解措施研究进展

运输应激是由动物经历长时间或长距离的异地运送所引起的机体非特异性反应,会严重扰乱动物生理状态、激素分泌及免疫抗氧化功能,严重时会危及动物生命。因此,缓解运输应激对机体造成的不利影响,对促进养殖业发展具有重要意义。文章综述运输应激对畜禽生理指标、内分泌激素、热休克蛋白、免疫、抗氧化及器官造成的不利影响,介绍营养代谢调控、改善肠道菌群、饲喂抗应激添加剂等缓解措施,从而为改善畜禽运输过程中健康状况及提高福利水平提供参考。