血清中不同甘露(聚)糖结合凝集素浓度的绵羊感染绵羊肺炎支原体后免疫因子表达的变化

为研究中国美利奴羊不同甘露(聚)糖结合凝集素(MBL)浓度感染绵羊肺炎支原体(MO)的免疫因子水平变化,本研究选择血清中MBL高、低浓度的绵羊各6只(感染组和对照组各3只),感染组人工感染MO,分别在人工感染前和感染后不同时间,采用荧光定量PCR法检测血液中血清因子及补体表达水平。结果显示,不同MBL浓度的绵羊人工感染后MBL m RNA水平呈下降趋势,MBL高浓度促炎因子IL-2和IFN-γ的m RNA表达水平较高,抗炎因子IL-4的m RNA表达水平在感染后1 d升高,此后开始下降,而IL-4的m RNA水平在14 d后有所升高;MBL低浓度羊感染后其TNF-α的m RNA水平显著升高,随炎症的缓解,逐渐降低;补体C1和C3的m RNA在感染后表现出不同的变化,MO感染可以激活补体途径。本研究结果表明,低血清MBL浓度与绵羊支原体肺炎具有一定的相关性,MBL不同浓度组之间其IL-2、IL-4、TNF-α、IFN-γ、补体C1、C3水平存在差异,低浓度MBL的绵羊更易发生比较严重的炎症反应。     

Effect of different levels of dietary sweet orange (Citrus sinensis) peel extract on humoral immune system responses in broiler chickens

This experiment was conducted to evaluate the effects of different levels of sweet orange (Citrus sinensis) peel extract (SOPE) on humoral immune system responses in broiler chickens. Three hundred 1‐day broilers (Ross‐308) were randomly allocated to treatments varying in supplemental SOPE added in the drinking water. The experimental groups consisted of three treatments fed for 42 days as follows: a control treatment without feed extract, a treatment containing 1000 ppm of SOPE and a treatment containing 1250 ppm of SOPE. All treatments were isocaloric and isonitrogenous. Broilers were vaccinated with Newcastle disease virus (NDV), avian influenza (AI), infectious bursal disease (IBD) and infectious bronchitis virus (IBV) vaccines. Antibody titer response to sheep red blood cells (SRBC) was higher in the group fed 1250 ppm of SOPE (P < 0.05) as well as for immunoglobulin G (IgG) and IgM. Similarly, antibody titer responses to all vaccines were constantly elevated (P < 0.05) by SOPE enrichment in a dose‐dependent manner. Relative weights of spleen and bursa of Fabricius were unaffected by treatments. Dietary SOPE supplementation may improve the immune response and diseases resistance, indicating that it can constitute a useful additive in broiler feeding. Thus, supplying SOPE in rations may help to improve relative immune response in broiler chickens.     

SCINTIGRAPHIC TRACKING OF MESENCHYMAL STEM CELLS AFTER PORTAL,SYSTEMIC INTRAVENOUS AND SPLENIC ADMINISTRATION IN HEALTHY BEAGLE DOGS

Mesenchymal stem cells have been proposed to treat liver disease in the dog. The objective of this study was to compare portal, systemic intravenous and splenic injections for administration of mesenchymal stem cells to target the liver in healthy beagle dogs. Four healthy beagle dogs were included in the study. Each dog received mesenchymal stem cells via all three delivery methods in randomized order, 1 week apart. Ten million fat‐derived allogeneic mesenchymal stem cells labeled with Technetium‐99m (99mTc)‐hexamethyl‐propylene amine oxime(HMPAO) were used for each injection. Right lateral, left lateral, ventral, and dorsal scintigraphic images were obtained with a gamma camera equipped with a low‐energy all‐purpose collimator immediately after injection and 1, 6, and 24 h later. Mesenchymal stem cells distribution was assessed subjectively using all four views. Pulmonary, hepatic, and splenic uptake was quantified from the right lateral view, at each time point. Portal injection resulted in diffuse homogeneous high uptake through the liver, whereas the systemic intravenous injection led to mesenchymal stem cell trapping in the lungs. After splenic injection, mild splenic retention and high homogeneous diffuse hepatic uptake were observed. Systemic injection of mesenchymal stem cells may not be a desirable technique for liver therapy due to pulmonary trapping. Splenic injection represents a good alternative to portal injection. Scintigraphic tracking with 99mTc‐HMPAO is a valuable technique for assessing mesenchymal stem cells distribution and quantification shortly after administration. Data obtained at 24 h should be interpreted cautiously due to suboptimal labeling persistence.     

枸杞多糖对雏鸡新城疫免疫效果的影响

为了研究枸杞多糖对雏鸡的免疫调节作用,将400羽1日龄雏鸡随机分为A、B、C、D共4组。7日龄时,分别给予A、B组雏鸡灌服10 mg/kg和20 mg/kg的枸杞多糖各5 m L,C、D组灌服同等剂量的生理盐水。给药的同时,A、B、C组滴鼻和点眼免疫新城疫Ⅳ系疫苗,D组不作处理。分别于7、14、21、28和35日龄时,采用微量法HI试验测定各组雏鸡新城疫抗体效价。分别于免疫后的第7、14、21、28和35天,计算各组雏鸡的免疫器官指数。试验结果显示,A、B组的新城疫抗体效价和各免疫器官指数明显高于C、D组,提示枸杞多糖对鸡新城疫疫苗的免疫效果具有明显的增强作用,且能够促进雏鸡中枢及外周免疫器官的发育。     

抗菌肽对发酵床饲养仔猪抗氧化指标、免疫指标及肠道主要菌群数量的影响

为研究发酵床饲养模式下饲粮中添加抗菌肽对仔猪生长性能、抗氧化指标、免疫指标及盲肠主要菌群数量的影响,选用108头35日龄,体重13kg左右的健康苏钟猪,随机分为3组,每组3重复,每重复12头猪,分别饲喂基础饲粮(对照组)、基础饲粮+40mg/kg杆菌肽锌+20mg/kg硫酸抗敌素(抗生素组)和基础饲粮+300 mg/kg抗菌肽(抗菌肽组),试验期49d。结果表明:(1)与对照组相比,饲粮中添加抗菌肽能够显著提高仔猪血清和肝脏中谷胱甘肽过氧化物酶活性(P0.05),极显著降低肝脏丙二醛含量(P0.01);抗菌肽组肝脏谷胱甘肽过氧化物酶活性比抗生素组极显著提高(P0.01)。(2)与对照组相比,饲粮中添加抗菌肽能够极显著提高仔猪十二指肠上皮内淋巴细胞数量(P0.01);抗菌肽组仔猪血清IgA含量分别比对照组和抗生素组提高4.03%、2.27%,抗菌肽组空肠上皮内淋巴细胞数量比对照组提高10.70%,但差异均不显著(P0.05)。(3)与对照组相比,饲粮中添加抗菌肽能够显著降低仔猪盲肠中大肠杆菌数量(P0.05);细菌总数降低5.98%,但差异不显著(P0.05)。综上所述,本试验条件下,饲粮中添加300mg/kg的抗菌肽能够提高仔猪抗氧化和免疫功能;减少仔猪盲肠中大肠杆菌数量,而对盲肠有益菌群无不良影响。     

凝结芽孢杆菌对感染产气荚膜梭菌肉鸡生长性能、肠道病变及免疫器官指数的影响

本试验旨在研究日粮中添加凝结芽孢杆菌对感染产气荚膜梭菌肉鸡生长性能、肠道病变及免疫器官指数的影响。选用312只1日龄科宝500肉鸡公雏,随机分成4个处理,每个处理6个重复,每个重复13只鸡。采用2×2双因子完全随机设计,产气荚膜梭菌分为无感染产气荚膜梭菌组和感染产气荚膜梭菌组,日粮中凝结芽孢杆菌水平分别为0和400mg/kg。试验期35d。结果表明,感染产气荚膜梭菌后15~35日龄肉鸡平均日增重显著降低(P0.05),28日龄肠道病变严重(P0.01),35日龄法氏囊指数极显著降低(P0.01);日粮中添加凝结芽孢杆菌可显著提高15~35日龄肉鸡平均日增重(P0.05),显著降低料重比(P0.05),能缓解感染导致的肠道损伤(P0.05),28日龄肉鸡胸腺指数极显著提高(P0.01),35日龄法氏囊指数极显著提高(P0.01)。肠道病变结果分析表明,日粮中添加凝结芽孢杆菌与感染产气荚膜梭菌有显著交互作用(P0.05)。综上所述,日粮中添加凝结芽孢杆菌能够提高感染产气荚膜梭菌肉鸡生长性能及免疫器官指数,增强动物免疫能力,并能缓解由于产气荚膜梭菌感染而导致的不利影响。     

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%.     

Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells

Background

There is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. Mesenchymal stem cells (MSCs) have been isolated and characterized from various species, but are poorly characterized in goats.

Results

Goat MSCs isolated from bone marrow (BM-MSCs) and adipose tissue (ASCs) have the ability to undergo osteogenic, adipogenic and chondrogenic differentiation. Cytochemical staining and gene expression analysis show that ASCs have a greater capacity for adipogenic differentiation compared to BM-MSCs and fibroblasts. Different methods of inducing adipogenesis also affect the extent and profile of adipogenic differentiation in MSCs. Goat fibroblasts were not capable of osteogenesis, hence distinguishing them from the MSCs. Goat MSCs and fibroblasts express CD90, CD105, CD73 but not CD45, and exhibit cytoplasmic localization of OCT4 protein. Goat MSCs can be stably transfected by Nucleofection, but, as evidenced by colony-forming efficiency (CFE), yield significantly different levels of progenitor cells that are robust enough to proliferate into colonies of integrants following G418 selection. BM-MSCs expanded over increasing passages in vitro maintained karyotypic stability up to 20 passages in culture, exhibited an increase in adipogenic differentiation and CFE, but showed altered morphology and amenability to genetic modification by selection.

Conclusions

Our findings provide characterization information on goat MSCs, and show that there can be significant differences between MSCs isolated from different tissues and from within the same tissue. Fibroblasts do not exhibit trilineage differentiation potential at the same capacity as MSCs, making it a more reliable method for distinguishing MSCs from fibroblasts, compared to cell surface marker expression.

Electronic supplementary material

The online version of this article (doi:10.1186/2049-1891-6-1) contains supplementary material, which is available to authorized users.     

Ultrastructural and Extracellular Protein Changes in Cell Suspension Cultures of Populus euphratica Associated with Low Temperature-induced Cold Acclimation

Populus euphratica Olive is the only tree species that can grow in the saline land and also survive cold winters in northwest China, and it plays a very important role in stabilizing the vulnerable ecosystem there. A cell suspension culture was initiated from callus derived from plantlets of Populus euphratica. Cold acclimation was induced (LT50 of-17.5 ℃) in cell suspension at 4-5 ℃ in the dark for 30 days and the freezing tolerance increased from LT50 of-12.5 ℃ in nonacclimated cells to LT50 of-17.5 ℃ in cold-acclimated cells. Microvacuolation, cytoplasmic augmentation and accumulation of starch granules were observed in cells that were cold-acclimated by exposure to low temperatures. Several qualitative and quantitative changes in proteins were noted during cold acclimation. Antibodies to carrot extracellular (apoplastic) 36 kD antifreeze protein did not cross react on immunoelectroblots with extracellular proteins in cell suspension culture medium of Populus euphratica, indicating no common epitopes in the carrot 36 kD antifreeze protein and P euphratica extracellular proteins. The relationship of these changes to cold acclimation in Populus euphratica cell cultures was discussed.     

油樟油细胞和粘液细胞发育的超微结构

利用超薄切片法和透射电镜研究油樟油细胞和粘液细胞发育过程。依据油细胞3层细胞壁的发育将其分为4个阶段。阶段1：仅有初生纤维素壁层，又可分为原始细胞和细胞液泡化二时期。质体内具白色小泡和黑色嗜饿滴，细胞质中有黑色或灰色的嗜饿物质，以及嗜饿物质与液泡的融合。阶段2：栓质化壁层的形成。片层状的栓质叠加在初生纤维素壁内侧。阶段3：内纤维素壁层的形成。较厚而结构松散的内纤维素壁层逐步形成，并叠加在栓质化壁层的内侧，大液泡成为充满嗜饿油脂的油囊。阶段4：油细胞成熟及细胞质解体。杯形构造由内纤维素壁层向细胞腔内突起形成，油囊由液泡包被连接到杯形构造中。解体的细胞质变得电子不透明或呈杂乱状态。粘液细胞发育方式有两种：一种是由内纤维素形成以后的油细胞发育而来，其细胞质中不断产生以同心圆或螺旋线方式排列的多膜结构，并充满整个细胞腔，最后多膜结构解体而成为丝状或颗粒状的粘液；另一种是由已完全成熟的油细胞发育而来，其油囊中的油呈不均匀的状态，并产生局部降解点，逐步扩大，最后油完全降解成颗粒成或丝状的粘液。