Evaluation of embryonic age and the effects of different proteases on the isolation and primary culture of chicken intestinal epithelial cells in vitro

The present study evaluates the effects of embryonic age and proteolytic enzymes on the isolation and primary culture of chicken enterocyte and to establish an effective technique for chicken intestinal epithelial cell (IEC) cultivation. Fourteen‐day‐old, 16‐day‐old and 18‐day‐old embryos (average weight: 52.23 ± 0.76 g, 50.86 ± 0.99 g, 48.98 ± 1.03 g) were the source for preparation of enterocyte culture, and trypsin‐ethylene diamine tetraacetic acid, collagenase, thermolysin and combination of collagenase and thermolysin were used for digestion medium. Optimal culture protocols were determined by qualitative assays of proliferation. Cells isolated by using 14‐day‐old embryo and collagenase obtain the best attachment and growth in culture, and the production of continuously growing IEC cultures. Thus, we conclude that the use of collagenase as a dissociating enzyme and 14‐day‐old embryo as a source can be advantageously applied to the isolation of chicken IEC and this method may be useful for various applications and basic studies of the intestinal tract concerning such objects as physiology, immunology and toxicology.     

Effects of Compound Chinese Medicine on Growth Performance,Nutrients Apparent Digestibility,Immune Function and Slaughter Performance of Weaning Rex Rabbits

This experiment was conducted to study the effects of different levels of compound Chinese medicine on growth performance,nutrients apparent digestibility,immune function and slaughter performance of weaning Rex rabbits.70 healthy Rex rabbits at age of 60 days were divided into 7 groups with 5 replicates per group and 2 rabbits per replicate.Group Ⅰ was fed with basic diet as control group,group Ⅱ was fed the basal diet supplemented with antibiotic (contains 50 mg/kg milligrams bacitracin zinc,10 mg/kg colistin sulfate).The other five groups (Ⅲ,Ⅳ,Ⅴ,Ⅵ and Ⅶ) were fed the basal diet supplemented with 1%,2%,3%,4%,5% compound Chinese medicine,respectively.The pre-test period lasted for 7 days and the trial period lasted for 45 days.The results showed as follows:①The ADG in group Ⅳ was significantly higher than that in groups Ⅰ,Ⅱ and Ⅲ (P< 0.05),the F/G in group Ⅳ was significantly lower than that in other groups (P< 0.05).②The apparent digestibility of DM in group Ⅳ was significantly higher than that in groups Ⅰand Ⅱ (P< 0.05),the apparent digestibility of DM in group Ⅶ was significantly higher than that in group Ⅰ (P< 0.05).The apparent digestibility of CF in group Ⅳ was extremely significantly higher than that in group Ⅰ (P< 0.01),the apparent digestibility of CF in group Ⅶ was significantly higher than that in group Ⅰ (P< 0.05).The apparent digestibility of NDF in group Ⅳ was extremely significantly higher than that in group Ⅰ (P< 0.01),and significantly higher than that in groups Ⅱ,Ⅲ,Ⅴ and Ⅶ (P< 0.05).The apparent digestibility of ADF in group Ⅳ was significantly higher than that in groups Ⅰ,Ⅱ,Ⅴ and Ⅶ (P< 0.05).The apparent digestibility of CP in group Ⅳ was significantly higher than that in groups Ⅰ and Ⅱ (P< 0.05).③ There were no significant differences in the contents of IgG,IgA and IgM of serum among all groups (P> 0.05),the spleen index in group Ⅳ was significantly higher than that in groups Ⅰ,Ⅱ,Ⅲ and Ⅴ (P< 0.05).④There were no significant differences in the slaughter rate,half eviscerated slaughter rate and all eviscerated slaughter rate among all groups (P> 0.05).In summary,adding compound Chinese medicine and antibiotics in the feed both could improve the growth performance,nutrients apparent digestibility,immune function and slaughter performance of weaning Rex rabbits,and the appropriate compound Chinese medicine level in the diet of weaning Rex rabbits was 2%.     

林龄、叶龄对亚热带杉木人工林碳氮稳定同位素组成的影响

[目的]研究5个不同林龄(3,8,14,21,46年)、不同叶龄(当年生、1年生、2年生、3年生)杉木人工林叶片的碳、氮稳定同位素组成,并根据它们对氮循环等过程的指示作用来探索不同林龄、叶龄杉木人工林氮循环过程及氮饱和程度的差异,从而为不同生长阶段杉木人工林制定施肥措施提供科学依据.[方法]以福建南平峡阳林场5块相互毗邻的不同林龄杉木人工林为研究对象,在每个林龄的林分内分别设置4个20 m ×20 m的试验小区.分别采集不同林龄杉木活叶并根据“主干法”将采集的杉叶分为不同叶龄,然后在每个小区内采集0 ～ 10 cm深度土层的土样,用同位素质谱仪测定它们的碳、氮稳定同位素组成(δ13C,δ15N),用碳氮元素分析仪测定叶片氮含量,叶片15N富集指数由叶片δ15N值减去相应的土壤δ15N得到.[结果]叶片δ15N值的变化范围为-2.52‰～2.81‰,叶片氮含量的变化范围为7.72％～13.5％,二者在不同林龄间均具有极显著差异,并均呈现出幼林和老林较高、处于速生期的林分较低的趋势,且叶片δ15N值与叶片氮含量之间存在显著的相关性,但不同叶龄叶片δ15N值间则不具有显著差异;不同林龄叶片的15N富集指数存在显著差异,呈现出幼林与老林叶片15N富集指数较接近于0的趋势;叶片δ13C的变化范围为-29.93‰～-27.88‰,不同林龄间差异不显著,但同一林龄不同叶龄叶片的δ13C则有显著差异,且有随着叶龄增大而减小的趋势.[结论]不同林龄叶片δ13C差异不显著但呈现幼年较低的趋势,可能是不同树高导致不同林龄杉木水分利用效率间的差异所致,而同一林龄不同叶龄杉木δ13C间的显著差异则可能是光合作用效率不同造成的.不同林龄在叶片δ15N、叶片氮含量、叶片15N富集指数间的显著差异均指示出处于速生期的林分氮饱和程度显著低于幼林和老林,这说明虽然我国亚热带地区氮沉降现象严重,但氮素仍是限制处于速生期杉木人工林生长的因素.     

荒漠植物白刺对模拟增雨的光合响应机制

【目的】研究不同增雨条件下白刺叶片的光合作用和叶绿素荧光特性,从光合生理角度探讨白刺对增雨的响应机制。【方法】以乌兰布和沙漠东缘地区典型荒漠植物白刺为研究对象,设置不同的增雨梯度(增加年均降水量的0,25%,50%,75%和100%),对自然生长的白刺沙包进行人工模拟增雨,利用 Li－6400xt便携式光合测定系统分析仪(USA,LI-COR)测定不同增雨条件下白刺叶片净光合速率(Pn)日动态变化、光响应曲线、CO2响应曲线以及叶绿素荧光参数,并根据 Pn 日变化曲线和响应曲线计算叶片日光合总量(∑Pn )以及各光合生理参数。【结果】1)75%和100%增雨对日均 Pn 和∑Pn 影响显著,日均 Pn 比 CK分别高32.74%和37.64%,∑Pn 比 CK 分别高32.01%和38.43%,说明增雨使白刺光合能力增强,日光合产物的积累增加。2)增雨使白刺表观量子效率( AQY)和光饱和点( LSP)升高,25%,50%,75%和100%增雨的 AQY 比 CK 分别高17.24%,31.03%,37.93%和24.14%,LSP比 CK分别高14.6%,6.0%,3.0%,26.1%,说明白刺叶片利用弱光的能力增强,对强光的利用范围增加,光能转化效率提高,有利于光合作用的高效进行。3)随着增雨量的增加,羧化效率(CE)呈现出逐渐增加的趋势,其中,100%增雨的 CE比 CK高5.73%;增雨使 CO2饱和点( CSP)升高,50%,75%和100%增雨的 CSP 显著高于 CK,分别比 CK高23.67%,28.35%和29.13%,说明白刺叶片 RuBP羧化酶和光合碳循环酶的活性增强,对高CO2浓度的利用范围增加。4)增雨使白刺叶片的原初光化学量子效率( Fv/Fm )、实际光化学量子效率(ФPSⅡ)、电子传递速率( ETR)和光化学猝灭系数( qP )提高,说明增雨有利于白刺叶片PSⅡ反应中心活性增强、开放比例提高,有利于叶片把所捕获的光能转化为生物化学能,并将更多的光能用于推动光合电子传递。【结论】白刺能够调节光合机构的功能、改变自身生理特性、增强对环境资源的利用能力来适应增雨的变化。     

三种野生观赏蕨类植物的耐旱性研究

本文研究狗脊、盾蕨、凤丫蕨的耐旱能力,为其园林应用提供参考依据。对三种蕨类植物材料在温室中采用自然干旱的方式进行干旱胁迫,每5天测定1次胁迫过程中生理生化指标,并测定其叶片保水力。结果表明:干旱胁迫下,狗脊的叶片保水力变化幅度最大,凤丫蕨其次,盾蕨最小;各植物电导率与对照组相比,均持续上升;叶绿素的含量均下降,凤丫蕨叶绿素含量变化幅度最大;叶片中的SOD活性、POD活性、MDA含量、可溶性蛋白含量变化趋势均为先升后降,凤丫蕨SOD活性、POD活性、MDA含量在前期迅速增长,狗脊、盾蕨缓慢上升。综合分析表明三种蕨类耐旱能力依次为狗脊盾蕨凤丫蕨。     

火烧对不同植被恢复类型物种多样性的影响

以山西省三交林场火烧迹地为研究对象,采用群落调查方法,分析了在自然更新下的不同植被恢复类型(油松人工林、灌木林、草本丛)下物种的组成及生物多样性。结果表明:(1)与对照相比,重度火烧后,草本丛的优势种重要值变化不明显,但喜阴耐旱的植物重要值显著升高;而灌木林主要以阳生耐旱草本植物占主要优势;油松人工林林分群落结构变化只是在灌木层和草本层中一些物种的更替。(2)重度火烧后所有植物的Simpson多样性指数均大于对照样地,油松人工林在重度火烧后多样性明显升高,油松人工林草本丛灌木林地。说明重度火烧2a后,不同类型植被恢复仍有一定的变化。     

Effect of reforestation on annual water yield in a large watershed in northeast China

A simplified water balance model in conjunc-tion with an evapotranspiration (ET) model and cumulative forest cover data were used to quantify the changes in annual water yield in response to reforestat...     

A new method to detect Lonsdalea quercina in infected plant tissues by real‐time PCR

Presymptomatic and accurate diagnoses of pathogens are essential for disease prediction and the timely application of bactericide. The bacterium Lonsdalea quercina (=Brenneria quercina) has been reported as the causal agent of drippy nut and bark canker disease on oak in California (US) and Europe. In recent years, it is also found on Populus × euramericana trees in Henan province of China. This bacterium causes longitudinal cankers of a few centimetres in size on the bark surface of the upper trunk. In this study, we developed two species‐specific PCR assays using primer pairs LqfF/LqfR and LqgF/LqgR for the rapid and accurate detection of the pathogenic bacteria in diseased plant tissues. The results show that the LqfF/LqfR primers amplified only a single PCR band of approximately 382 bp and the LqgF/LqgR primers yielded a PCR product of approximately 286 bp. The two primers were successfully adapted to real‐time PCR based on SYBR Green I used with the ABI 7500 system. The detection limit of the reaction was 0.1 pg genomic DNA per 20 μl PCR reaction volumes. The pathogen was mainly detected in the phloem of cankers as well as in the exudates of diseased trees, but was not found in the xylem or leaves. The size of pathogen in distribution was larger than the lesion. The results demonstrate that real‐time PCR assays can be used to detect the pathogen by extracting DNA directly from infected plant tissues. This method is a rapid, reliable method for the presymptomatic and accurate detection of L. quercina, providing a useful insight into epidemiological studies.     

膜下滴灌结合暗管排水技术改良新疆盐碱地效果综述

膜下滴灌技术的大面积推广和应用,为新疆乃至西北干旱地区作物高产提供了强大的技术支撑。应用在盐碱地改良中,膜下滴灌“有灌无排”模式仅调节耕层盐分分布,并不能将盐分排出土体,长年灌溉容易引起爆发式积盐的风险,而暗管排水是目前公认的改良盐碱地最直接、最有效的方法。目前,将膜下滴灌与暗管排水有机结合形成的新型灌溉技术,充分利用膜下滴灌良好的抑蒸增温、节水抑盐等优势,同时利用暗管将淋洗盐水及时排出土体,使盐碱化土层产生新的水盐平衡,已在新疆盐碱地区开发利用中得到长足发展。本文重点分析并总结了新疆灌区膜下滴灌结合暗管排水系统的设计参数及土壤水盐运移规律,并综述了土壤理化性质及作物产量对该灌排技术的响应研究,以期为新疆盐碱地农业可持续发展提供理论依据和技术指导。     

Dietary dihydroartemisinin supplementation alleviates intestinal inflammatory injury through TLR4/NOD/NF-κB signaling pathway in weaned piglets with intrauterine growth retardation

The aim of present study was to evaluate whether diets supplemented with dihydroartemisinin (DHA) could alleviate intestinal inflammatory injury in weaned piglets with intrauterine growth retardation (IUGR). Twelve normal birth weight (NBW) piglets and 12 piglets with IUGR were fed a basal diet (NBW-CON and IUCR-CON groups), and another 12 piglets with IUGR were fed the basal diet supplemented with DHA at 80 mg/kg (IUGR-DHA group) from 21 to 49 d of age. At 49 d of age, 8 piglets with similar body weight in each group were sacrificed. The jejunal and ileal samples were collected for further analysis. The results showed that IUGR impaired intestinal morphology, increased intestinal inflammatory response, raised enterocyte apoptosis and reduced enterocyte proliferation and activated transmembrane toll-like receptor 4 (TLR4)/nucleotide-binding and oligomerization domain (NOD)/nuclear factor-κB (NF-κB) signaling pathway. Dihydroartemisinin inclusion ameliorated intestinal morphology, indicated by increased villus height, villus height-to-crypt depth ratio, villus surface area and decreased villus width of piglets with IUGR (P < 0.05). Compared with NBW piglets, IUGR piglets supplemented with DHA exhibited higher apoptosis index and caspase-3 expression, and lower proliferation index and proliferating cell nuclear antigen expression in the intestine (P < 0.05). Dihydroartemisinin supplementation attenuated the intestinal inflammation of piglets with IUGR, indicated by increased concentrations of intestinal inflammatory cytokines and lipopolysaccharides (P < 0.05). In addition, DHA supplementation down-regulated the related mRNA expressions of TLR4/NOD/NF-κB signaling pathway and upregulated mRNA expressions of negative regulators of TLR4 and NOD signaling pathway in the intestine of piglets with IUGR (P < 0.05). Piglets in the IUGR-DHA group showed lower protein expressions of TLR4, phosphorylated NF-κB (pNF-κB) inhibitor α, nuclear pNF-κB, and higher protein expression of cytoplasmic pNF-κB in the intestine than those in the IUGR-CON group (P < 0.05). In conclusion, DHA supplementation could improve intestinal morphology, regulate enterocyte proliferation and apoptosis, and alleviate intestinal inflammation through TLR4/NOD/NF-κB signaling pathway in weaned piglets with IUGR.