Fat depot‐specific differences in leptin mRNA expression and its relation to adipocyte size in steers

ABSTRACT This experiment was conducted to investigate leptin mRNA expression, adipocyte size, and their relationship in several adipose tissues of fattening steers. Subcutaneous, perirenal, intermuscular and intramuscular adipose tissues were collected from three crossbred steers (Japanese Black cattle X Holstein) aged 21 months. The mRNA level and adipocyte diameter were determined in these adipose tissues. The intramuscular adipose tissue had a lower leptin mRNA level than the intermuscular and perirenal adipose tissues (P < 0.05). Leptin mRNA level was lower in the subcutaneous depot than in the intermuscular depot (P < 0.05). Adipocyte diameter was larger in the intermuscular adipose tissue than in the subcutaneous and intramuscular adipose tissues (P < 0.05). Leptin mRNA level was positively correlated with adipocyte diameter (r² = 0.81, P < 0.05). These results suggest that the cattle have fat depot‐specific differences in leptin gene expression, which are a result of a difference in adipocyte size.     

Development and characterization of mouse monoclonal antibodies reactive with chicken CD83

This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD83 (chCD83), a membrane-bound glycoprotein belonging to the immunoglobulin superfamily that is primarily expressed on mature dendritic cells (DCs). A recombinant chCD83/IgG4 fusion protein containing the extracellular region of chCD83 was expressed in Chinese Hamster Ovary (CHO) cells and isolated from the spent cell culture medium by protein G affinity chromatography. The extracellular region of the chCD83 protein was purified and used to immunize mice. A cell fusion was performed, from which 342 hybridomas were screened for mAbs to chCD83. Two mAbs, chCD83-159 and chCD83-227, stained the greatest percentage of chCD83-transfected CHO cells and were selected for further characterization. By flow cytometry, both mAbs reacted with a chicken macrophage cell line, HD11. Both mAbs also recognized a single 53 kDa protein on Western blots of lysates from lipopolysaccharide-stimulated spleen mononuclear cells or unstimulated HD11 cells. Immunostaining of chicken secondary lymphoid organs identified chCD83(+) cells with morphologic and subtissue localization properties comparable to mammalian DCs. In vitro stimulation of spleen mononuclear cells with concanavalin A (Con A) decreased the percentage of chCD83(+) cells compared with cells treated with medium alone. Interestingly, spleen cells treated with Con A in the presence of chCD83-227 mAb exhibited decreased percentage of MHCII(+) cells compared with cells treated with an isotype-matched negative control mAb. These chCD83 mAbs may be useful for future investigations of chicken immune cell maturation and mechanisms of action.     

The effects of replacement of antibiotics with by‐products of oriental medicinal plants on growth performance and meat qualities in fattening pigs

The effect of by‐products of oriental medicinal plants (OMP; T1) containing 0.03% herb extracts (T2) or 0.1% aminolevulinic acid (T3) on the production performance of swine during the finishing period and on its meat quality were investigated. No significant differences were found in the weight gain, feed intake and feed conversion rate among the tested groups (P > 0.05). But the treated group showed higher (P < 0.05) moisture and ash and lower protein than the control group. The T3 group showed a lower meat cholesterol content (38.42 mg/100 g) compared to the other groups (P < 0.05). The vitamin E content of the muscle in the treated groups was higher compared to the control group. No antibiotic content was detected in all treated and control samples. The values of the volatile basic nitrogen (VBN) and thiobarbituric acid reactive substance (TBARS) of the treated groups were significantly lower (P < 0.01) than the control group. The treated groups had significantly better (P < 0.05) sensory‐test scores for color, flavor, off‐flavor and total acceptability compared to the control group.     

狐狸源致病性维氏气单胞菌的分离鉴定及耐药性分析

本研究通过生物学特性、16S rRNA基因和4种等看家基因的序列测定与分析,对分离自北京动物园死亡狐狸病变组织的1株细菌进行鉴定,并对其进行小鼠致病力试验及药物敏感性试验。结果显示:该细菌为革兰氏阴性杆菌,培养特性、理化反应与维氏气单胞菌温和生物型基本相同;根据16S rRNA基因和dnaJ、rpoD、gyrB、cpn60等看家基因的系统进化及其同源性分析,确定所分离的细菌与维氏气单胞菌属于同一系统发育分支,各基因同源性最高分别为97.6%～99.9%。分离菌株对CD-1小鼠有较强的致病作用,其半数致死量为2.8×105cfu/只;对氧氟沙星、头孢他啶等14种抗菌药物敏感,对青霉素G、克林霉素等9种抗菌药物耐药。     

Molecular characterization of tetracycline- and quinolone-resistant Aeromonas salmonicida isolated in Korea

The antibiotic resistance of 16 Aeromonas (A.) salmonicida strains isolated from diseased fish and environmental samples in Korea from 2006 to 2009 were investigated in this study. Tetracycline or quinolone resistance was observed in eight and 16 of the isolates, respectively, based on the measured minimal inhibitory concentrations. Among the tetracycline-resistant strains, seven of the isolates harbored tetA gene and one isolate harbored tetE gene. Additionally, quinolone-resistance determining regions (QRDRs) consisting of the gyrA and parC genes were amplified and sequenced. Among the quinolone-resistant A. salmonicida strains, 15 harbored point mutations in the gyrA codon 83 which were responsible for the corresponding amino acid substitutions of Ser⁸³→Arg⁸³ or Ser⁸³→Asn⁸³. We detected no point mutations in other QRDRs, such as gyrA codons 87 and 92, and parC codons 80 and 84. Genetic similarity was assessed via pulsed-field gel electrophoresis, and the results indicated high clonality among the Korean antibiotic-resistant strains of A. salmonicida.     

高山草原条件下苜蓿种质抗霜霉性评价

对引自国内外的94个苜蓿品种在海拔3000m的甘肃桑科摹试验站进行了高山草原条件下的霜霉病抗性评价。结果表明,品种之间的抗病性差异颇大,发病率为0～100%,严重度为1～4。通过对各品种的发病率和严重两项指标进行聚类分析,按照抗病性从高到低将94个品种分为免疫、高抗、中抗、抗病、感病、中感、高感和极感8个抗性类群;其中：阿尔古奎斯、巴瑞尔,阿毕卡、安古斯、日本1品种、伊鲁瑰斯、布韦兹、托尔、81-89美国、CP4350萨蓝纳斯、贝维、润布勒、兰热来恩德和威斯康辛等14个品种属免疫类群;普劳勒、班纳、L2-1079匈牙利和兴平4个品种属高抗类群;肇东、陇中和陇东3个品种属高感类群;78-27捷克、阿波罗、准格尔、陕北和河西5个品种属极感类群。6～8月份病害随着生长期的推移而呈逐渐加重的趋势。绝大多数极感品种和所有高感品种均为国内品种,而所有免疫品种和绝大多数高抗品种均为国外引进品种,这种由于品种的来源不同而引起的抗病性差异很可能与苜蓿霜霉菌的高度寄生专化性有关。     

狗牙根果岭的交播技术

主要介绍狗牙根果岭的交播技术 ,包括草种选择和配比、种子直播、交播前后的管理措施 ,并讨论了狗牙根果岭的春季过渡技术等。     

Determination of Feeder Cell‐Based Cellular Niches Supporting the Colonization and Maintenance of Spermatogonial Stem Cells from Prepubertal Domestic Cat Testes

Recently, isolation and in vitro culture of putative spermatogonial stem cells (SSCs) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long‐term maintenance of feline SSCs (FSSCs) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSCs among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells (TSCs) derived from cats, dogs and mice, and mouse embryonic fibroblasts (MEFs). The formation and growth of colonies derived from SSCs cultured on each type of feeder cell were identified at passage 0, and the morphology, alkaline phosphatase (AP) activity and expression of SSC‐specific genes in surviving colonies were investigated at passage 4. Among these diverse feeder cells, TSCs from cat showed the greatest colony formation, growth and maintenance of FSSCs, and SSC colonies cultured by passage 4 showed a typical dome‐shaped morphology, AP activity and expression of SSC‐specific genes (NANOG, OCT4, SOX2 and CD9). Accordingly, these results demonstrate that feline TSCs could be used as feeder cells to support the establishment and maintenance of SSCs from domestic cats.     

苜蓿草粉对禾花鲤蛋白酶和淀粉酶活性影响的研究

选取来源一致、规格整齐的2龄禾花鲤1 200尾,采用单因子完全随机设计,分为5个处理,每处理3个重复,每重复80尾鱼,分置于15个试验池中。各处理苜蓿Medicago sativa草粉的添加量分别为0（对照组）、40（试验Ⅰ组）、80（试验Ⅱ组）、120（试验Ⅲ组）和160 g/kg（试验Ⅳ组）,正试期50 d,观测苜蓿草粉对禾花鲤前、中、后肠及肝胰脏蛋白酶和淀粉酶活性的影响。结果表明：1）添加苜蓿草粉对前肠蛋白酶活性有显著影响 (P<0.05),其中试验II组显著高于对照组和试验Ⅳ组,但和试验Ⅰ、Ⅲ组差异不显著 (P>0.05)。添加苜蓿草粉对中、后肠及肝胰脏蛋白酶活性均无显著影响 (P>0.05),但它们有相同的变化趋势,即试验Ⅰ、Ⅱ、Ⅲ组鱼的蛋白酶活性均高于对照组。2）鲤鱼的前肠、中肠、后肠蛋白酶活性依次减弱,而肝胰脏的蛋白酶活性远低于肠道。3）添加苜蓿草粉后前肠及中肠淀粉酶活性比较,试验II组显著高于对照组 (P<0.05),其余各组与对照组差异不显著 (P>0.05),后肠淀粉酶活性与对照组无显著差异 (P>0.05)。添加苜蓿草粉能提高鲤鱼肝胰脏淀粉酶活性,其中试验Ⅱ组显著高于对照组 (P<0.05),其余试验组与对照组无显著差异 (P>0.05)。4）禾花鲤前、中、后肠及肝胰脏4部分淀粉酶的活性有较大的不同,肝胰脏>后肠>中肠>前肠。     

牛消化道Ⅰ型肽载体克隆测序及编码蛋白结构分析

从牛瓣胃上皮提取RNA,根据已知的人、鼠、兔、羊PepT1序列设计引物,PCR扩增目的片段cDNA,扩增产物经纯化、克隆后测序.克隆测序片段为1566 bp(DQ309694),与其它动物已知PepT1序列比对,发现该片段位于第3～10跨膜结构域之间.牛测定序列与人、鼠、兔、羊、狗、鸡和猪PepT1相应片段核苷酸序列同源性分别为82.89%、80.14%、78.93%、96.04%、83.08%、63.90%和85.66%,相应片段氨基酸序列同源性分别为81.45%,79.62%、76.25%、94.06%、81.49%、61.41%和83.56%.对8种动物(牛、人、鼠、兔、羊、狗、鸡、猪)测序片段内氨基酸序列同源性分析表明,PepT1是高度保守的蛋白,其中跨膜结构域的氨基酸序列保守性高于膜外环.8种动物PepTl测序片段内各跨膜结构域的氨基酸序列同源性均在90%以上,第3、4、 5 6、7,8,9和10跨膜结构域的氨基酸序列同源性分别为90.97%、91.07%、97.62%、93.45%、94.05%、91.67%、91.45%和95.83%.8种动物测序片段内膜外环的氨基酸序列同源性在70%～99%之间,以第4-5跨膜结构域之间膜外环的同源性最高,9-10跨膜结构域之间膜外环的同源性最低,3-4、4-5、5-6,6-7、7-8、8-9、9-10跨膜结构域之间膜外环的氨基酸序列同源性分别为77.63%、98.86%、86.72%、90.13%、91.25%、88.33%和71.74%.9-10结构域之间是1个位于细胞外的大膜外环,可能对底物的识别有重要作用,牛在此区域与鸡相同区域的氨基酸序列同源性仅为29.3%,与大鼠,狗、人、兔、猪和羊相应序列的同源性依次为65.17%、71.64%、67.16%、60.20%、73.13%和88.56%.对测序BPepT1片段编码蛋白的糖基化和磷酸化位点分析表明,测序片段编码蛋白共有6个N-糖基化位点和3个蛋白激酶C依赖性(PKC)磷酸化位点.