无抗发酵饲料对生长育肥猪生长性能、肠道菌群和养分表观消化率的影响

本试验旨在研究无抗发酵饲料对生长育肥猪生长性能、肠道菌群和养分表观消化率的影响。选择18 kg左右断奶仔猪60头,随机分为2个组,每组3个重复,每个重复10头猪。对照组饲喂含抗生素基础饲粮(每千克含黄霉素5 mg),试验组饲喂80%基础饲粮和20%的无抗发酵饲料组成的试验饲粮。试验期154 d。结果表明:1)试验组的末重、平均日增重和平均日采食量都显著高于对照组(P0.05),而料重比则显著低于对照组(P0.05)。2)试验组中期和后期的乳酸菌数量的对数值显著高于对照组(P0.05),而试验组中期和后期的大肠杆菌和沙门氏菌数量的对数值显著低于对照组(P0.05)。3)在十二指肠肠段,试验组肠道食糜中的乳酸菌和大肠杆菌数量的对数值显著高于对照组(P0.05)。4)在十二指肠肠段,试验组肠道每克和每平方厘米黏膜的乳酸菌数量的对数值显著高于对照组(P0.05);试验组肠道每克黏膜的大肠杆菌数量的对数值显著高于对照组(P0.05)。在空肠肠段,试验组肠道每克和每平方厘米黏膜的乳酸菌数量的对数值显著高于对照组(P0.05)。5)试验组的粗蛋白质和粗纤维的表观消化率显著高于对照组(P0.05)。由此可见,无抗发酵饲料能提高生长育肥猪的生长性能,改善肠道微生物平衡,增强消化能力。     

肠道菌群对仔鸡肠道发育、黏膜形态和免疫器官发育的影响

本试验旨在以"无菌鸡"为模型,研究添加鸡源性乳酸菌和盲肠内容物后,对仔鸡肠道发育、黏膜形态及免疫器官发育的影响。试验选用无特定病原体(SPF)种蛋80枚,经无菌孵化后,选择健康仔鸡60羽,分3组(每组20羽),转入3台无菌隔离器内饲养,并分别饲喂:无菌饲粮(CT组)、无菌饲粮+乳酸菌液(LB组)、无菌饲粮+鸡盲肠内容物(CC组);饲养21 d后试验结束,于试验7、14、21日齡从各组取6只进行屠宰,测定鸡肠道各段长度和免疫器官重量,并对肠道黏膜形态显微观察。结果显示:饲喂乳酸菌或盲肠内容物的仔鸡各阶段中的空肠长度均高于CT组,盲肠长度和体积均低于CT组的无菌鸡;仔鸡的脾脏指数高于CT组(P=0.010);饲喂乳酸菌或盲肠内容物后仔鸡的十二指肠、空肠和回肠隐窝深度降低、绒毛高度/隐窝深度增大。本试验说明,饲喂肠道菌群能促进仔鸡肠道发育,盲肠体积变小;同时促进脾脏的发育,提高机体免疫力。     

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.     

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.     

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。     

Comparison Study on Effects of Three Chinese Herbal Medicine on Intestinal Morphology in Weaning Piglets

The objective of this study was to compare the effect of three Chinese herbal medicine (medicated leaven, hawthorn, fructus hordei germinates) on histomorphology of different parts of small intestine in weaning piglets.40 28-day old ternary pigs (Duroc×Landrace×Yorkshire) with the body weight (8.13±1.32)kg were divided into four groups, control group, medicated leaven group, hawthorn group and fructus hordei germinates group, and feeding with 1% Chinese herbal medicine, there were two repeats in each group and each repeat had 5 piglets, slaughtering and sampling at 7th and 14th day morning, respectively, sections were used for HE staining.The staining tissues were taken pictures under a microscope camera (200×) and measured VH, CD, IWT and V/C in different parts of small intestine with Image-Pro Plus.The result showed that the IWH of ileum in medicated leaven group was significantly or extremely significantly higher than control group (P<0.05;P<0.01).The VH and IWT of duodenum and jejunum at 7th day in hawthorn group were extremely significantly lower than control group (P<0.01);The IWT of ileum and the VH of ileum at 14th day were extremely significantly higher than control group (P<0.01).In fructus hordei germinates group, the VH of duodenum and jejunum at 7th day and IWT of jejunum were extremely significantly lower than control group (P<0.01);The CD of ileum at 7th day, the IWT of duodenum and ileum at 7th day were extremely significantly higher than control group (P<0.01).The effects of different simple Chinese herbal medicine on histomorphology of different parts of small intestine in piglets were different, because of the different digestive functions of small intestine, the most effects of Chinese herbal medicine an duodenum, jejunum and ileum were fructus hordei germinates, medicated leaven and hawthorn, respectively.     

Effect of Soybean Meal Replaced by Brewer's Grain or Fermented Brewer's Grain on Growth Performance,Immune Organ Index and Intestinal Flora of Meat Geese

This experiment was conducted to investigate the effect of soybean meal replaced by brewer's grain (BSG) or fermented brewer's grain (FBSG) on growth performance,immune organ index and intestinal flora of meat geese.A total of 250 geese at 28 days of age were randomly assigned to 5 groups with 5 replicates per group and 10 geese per replicate.The geese in control group were fed a basal diet (with soybean);The geese in groups 1 and 2 were fed the basal diet with the 10% and 20% BSG replacing the soybean,respectively;The geese in groups 3 and 4 were fed the basal diet with the 10% and 20% FBSG replacing the soybean,respectively.The whole experiment period was 42 days,including three phases:15 d (early stage),14 d (middle stage) and 14 d (later stage).The results showed as follows:①From the whole experiment period,compared with control group,the ADG of geese in group 1 was significantly increased (P<0.05),the ADG and ADFI of geese in group 4 were significantly decreased (P<0.05),there were no obvious change in growth performance of geese in groups 2 and 3 (P>0.05).② Compared with control group,the thymus index and spleen index of geese in groups 1,2 and 3 were extremely significantly increased at early stage (P<0.01);At middle and later stages,the spleen index of geese in group 2 were significantly and extremely significantly increased (P<0.05;P<0.01);There were no significant differences in bursa of fabricius index of geese among all groups (P>0.05).③ Compared with control group,the number of Escherichia coli and Salmonella in cecum in groups 1 and 2 were extremely significantly decreased at early and later stages (P<0.01),while the number of Bifidobacteria and Lactobacillus were increased in different degrees;the number of Bifidobacteria and Lactobacillus in cecum in groups 3 and 4 were extremely significantly increased at later stage (P<0.01).In conclusion,10% BSG replacing soybean in the basal diet could improve the growth performance and intestinal flora of meat geese;10% FBSG replacing soybean in the basal diet could promote the growth of beneficial bacteria of meat geese,but had no effect on growth performance;While 20% replacing soybean in the basal diet could decrease the feed intake and weight gain of meat geese,hinder its growth and development.     

Effect of Heat Shock Protein-90 (HSP-90) mRNA Expression in Intestinal Tissuse of Chickens Infected with E.necatrix

The 90 chickens were randomly divided into control group, E.necatrix one time infection group and E.necatrix two times infection group.We used duplex PCR method to test the expression changes of HSP-90 mRNA in intestinal tissue of chickens infected with E.necatrix and researched the effects of it comprehensively and systematically.The results were showed that after 0 to 14 days the expression of HSP-90 mRNA in intestinal tissues of 14-day-old chickens infected with E.necatrix presented a upward trend though it was lower than the control chickens.These chickens were infected with E.necatrix again at 28 days, HSP-90 mRNA expressions of duodenum, jejunum and ileum decreased and then increased.However, it always showed a downward trend in appendix.When 28 days chickens were infected with high-does E.necatrix once, HSP-90 mRNA expressions of duodenum, jejunum and appendix were in a declining curve.While it was increased originally and then decreased in ileum.This study indicated that HSP-90 mRNA expressions in intestinal tissue of chickens infected with E.necatrix were inhibited at first, and the expression quantity was closely related to the degrees of intestinal tissue damage.     

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.