广西沼泽型水牛IL-18和TNF-α基因的克隆与序列分析

从健康沼泽型水牛静脉无菌采血,分离外周血单核细胞(PBMCs),用刀豆素A(ConA)诱导培养3,8和12 h后,提取细胞总RNA,以Oligo(dT)18为引物合成第一链cDNA,根据牛的白细胞介素18(IL-18)和绵羊的肿瘤坏死因子α(TNF-α)基因序列设计2对特异性引物,通过PCR方法扩增出水牛IL-18和TNF-α基因,将其克隆到pMD18-T载体,测序后进行序列分析。结果试验中所克隆到的IL-18和TNF-α基因序列的开放阅读框(ORF)分别为602 bp和715 bp,与GenBank所登录的水牛IL-18和TNF-α核苷酸序列同源性为99.1%和99.7%,其推导的氨基酸序列同源性分别是99.0%和98.7%。表明成功从水牛外周血中克隆到了水牛IL-18和TNF-α基因。     

Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

We previously reported that the release of O₂ ⁻ from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. The nucleotide sequence data of PsCu/Zn-SOD1 reported are available in the DDBJ/EMBL/GenBank databases under accession number AB189165.     

白耳黄鸡生产性能和肉质性状的研究

对白耳黄鸡生长性能、屠宰性能、常规肉质、胸腿肌化学成分、肌肉组织学特性、胸肌氨基酸和脂肪酸含量等指标进行了研究。白耳黄鸡公母鸡的屠宰率、胸肌率、腿肌率、蛋白质含量、氨基酸总量、腹脂率、人体必需氨基酸含量、必需脂肪酸含量等分别为89.23/90.18%,16.65/18.09%,22.37/21.91%,23.64/25.30%,206.29/218.8mg/g,1.64/2.39%,92.78/96.75mg/g,30.69/29.52%。结果表明,白耳黄鸡的各指标都表现优良,其具有屠宰性能好,肉质优良,必需氨基酸和脂肪酸含量高,腹脂率低等特点,是一个值得推广的地方优良品种。     

NO、TNF和IL-2与新型鸭肝炎病毒感染雏鸭肝脑组织损伤的关系

用新型鸭肝炎病毒人工感染9日龄健康樱桃谷雏鸭，对感染后12、24、48、96、168h和14d雏鸭血液、肝和脑组织中一氧化氮（NO）含量，血液中肿瘤坏死因子（TNF）和白细胞介素2（IL-2）含量进行了测定，同时对感染雏鸭的组织病理学变化进行了观察。结果表明．血清中NO含量在接种后48h开始升高，一直持续到接种后96h，接种后7d恢复正常；肝脏组织中NO含量仅在接种后24h与对照组比较显著升高，在其他时间未表现有差异；脑组织中NO含量在整个试验期间没有变化。血清中的TNF和IL-2含量在接种后24h均表现升高，接种后96h降低，其他时间无改变。感染雏鸭的肝组织在接种后24h表现出血性坏死性肝炎变化，接种后48～96h呈增生性病变，而接种后各时期脑组织均呈非化脓性脑炎变化。由此表明，新型鸭肝炎病毒感染可导致雏鸭体内NO、TNF和IL-2发生变化，并且与肝组织损伤、疾病的发生发展有关。     

牛GDF9和BMP15基因遗传变异与双胎性状的关系研究

以生长分化因子9(Growth differentiation factor 9,GDF9)基因和骨形态发生蛋白15(Bone morphogenet-ic protein 15,BMP15)基因作为牛双胎性状的候选基因,研究了它们在鲁西牛、秦川牛、南阳牛和中国荷斯坦牛4个品种中的遗传变异,并在鲁西牛群体中研究了其多态位点与双胎性状的关系。结果表明:在鲁西牛中GDF9基因的3′UTR发现缺失突变,而其它3个品种中没有发现该突变。对鲁西牛群体中该多态位点与单、双胎性状之间进行卡方显著性检验表明,单胎牛群体与双胎牛群体基因型分布有极显著的差异(P=0.006),双胎牛群体的B等位基因频率明显大于单胎牛群体。通过生物信息学分析表明,突变体mRNA的二级结构与野生型相比总自由能值差异不大,但突变体mRNA翻译起始位点的二级结构稳定性明显大于野生型。在鲁西牛、南阳牛和秦川牛的BMP15基因中发现编码区第759~762位有GAAA 4个碱基存在缺失突变,但没有检测到突变纯合个体,中国荷斯坦牛中没有检测到该突变。卡方显著性检验表明单胎牛群体和双胎牛群体在该位点基因型组成差异不显著(P=0.947)。     

猪IL-2真核表达质粒构建及对PRRSV-ORF5基因疫苗免疫佐剂作用的研究

本研究应用真核表达载体pcDNA3.1（＋）和猪白细胞介素2（IL-2）基因成功构建了IL-2的真核表达质粒（pcDNA-IL-2）,探讨了pcDNA-IL-2作为分子免疫佐剂对猪繁殖与呼吸综合征（PRRS）ORF5基因疫苗（pcDNA-PRRSV-SC2-ORF5）免疫猪的增强作用。经间接ELISA法、MTT比色法及流式细胞仪分别对pcDNA-IL-2与pcDNA-PRRSV-SC2-ORF5共同免疫猪、pcDNA-PRRSV-SC2-ORF5单独免疫猪、pcDNA3.1（＋）空载体和灭菌水免疫对照猪的血清抗PRRSV抗体IgG、外周血T淋巴细胞的增殖活性、CD4^＋和CD8^＋细胞比例进行检测,结果表明pcDNA-IL-2与pcDNA-PRRSV-SC2-ORF5共同免疫猪的IgG含量、外周血T淋巴细胞的增殖活性、CD4^＋和CD8^＋细胞比例与pcDNA-PRRSV-SC2-ORF5单独免疫猪相比有显著差异（P〈0.05）。说明pcDNA-IL-2能显著增强pcDNA-PRRSV-SC2-ORF5基因疫苗免疫猪的体液和细胞免疫应答,可作为PRRSV基因疫苗的良好佐剂。     

草莓对不同形态氮素的吸收反应

以鬼怒甘草莓为试材,研究草莓对铵态氮与硝态氮的吸收反应.结果表明,水培务件下草莓对铵态氮的吸收利用率比硝态氮高2倍以上,同时供应铵态氮和硝态氮,比单独使用铵态氮或硝态氮的利用率高.草莓对不同形态氮素的反应与栽培方式有关,露地草莓根、茎、叶的生物量随着铵态氮比例增加而降低,而温室草莓的结果与此相反;露地栽培条件下以硝态氮/铵态氮为5:2时产量最高,冬季温室栽培条件下以硝态氮/铵态氮为2:5时产量最高.     

Using a tobit regression model to analyse risk factors for foot-pad dermatitis in commercially grown broilers

A model was developed to explain the influence of different local and regional factors on the prevalence of foot-pad dermatitis in broilers at the time of slaughter. Data from a recent two-year study of broiler foot-health in Sweden were used to construct the model. The model was based on regression analysis, using a Tobit model for censored data. This type of Tobit regression model has been used for several decades in econometrics and can also be found in the medical-scientific literature. It has, however, reportedly not been used in veterinary medicine although there appears to be several situations where it should be considered useful. Week of slaughter was significantly (p<0.001) associated with total foot-pad score, which decreased over time. The birds' age at slaughter was also significantly (p<0.01) associated with total foot-pad score. Five out of the 11 slaughterhouses included in the study were included as significant (p<0.01) variables in the final model. Three feed suppliers (p<0.05) and four regions (p<0.01) were also included as significant variables in the model. There were also significant (p<0.001) interactions between several of these factors (mainly, between the time variable and specific slaughterhouses, feed suppliers or regions, respectively).

The information gathered in this study regarding the influence of a number of risk factors for the development of foot-pad dermatitis and their reciprocal interactions will enable us to modify the Swedish broiler foot-health programme.     

Can hydrochar and pyrochar affect nitrogen uptake and biomass allocation in poplars?

Positive effects of pyrochar on soil nutrient availability and plant growth are widely reported in literature. However, few studies have reported effects of hydrochars on plant nutrition. A pot trial was conducted over a period of 2 years to investigate the effect of a pyrochar (AGT) and a hydrochar (HTC) on poplar (Populus × generosa , clone AF8) growth, biomass allocation and nitrogen (N) uptake with special emphasis on the quantification (using an isotopic mass balance approach) of char‐derived nitrogen (CDN) absorbed by plants. We found that both pyrochar and hydrochar positively affected above‐ground biomass productivity in the first year, and biomass and nitrogen (N) allocation over the 2 years by reducing the allocation of resources to fine roots. By the end of the experiment, even though the total N uptake was not affected by char, the CDN was more than 24% of the total N absorbed by HTC‐treated plants compared to a negligible amount absorbed by AGT‐treated ones. Finally, char did not affect nitrogen use efficiency (NUE) in the first year of growth, but by the end of the experiment, NUE was higher in the above‐ground biomass of HTC‐treated than in AGT‐ and control poplars.     

培养液中铜水平对PK15细胞内铜离子浓度与CuZn-SOD活性的影响

在 PK15细胞无血清液体培养基中添加 0、7.8、15 .6、31.2、6 2 .5 μmol/L Cu,观察了培养液中不同铜水平对PK15细胞内铜离子浓度和 Cu Zn- SOD活性的影响及二者的相关性。结果表明 ,PK15细胞内铜离子浓度及 Cu Zn-SOD活性均随培养液中添加铜离子浓度升高和培养时间的延长而有不同程度的升高 ,其中以添加 Cu31.2 μmol/L组最高 (P<0 .0 5 )。PK15细胞内 Cu Zn- SOD活性与细胞内铜离子浓度和培养时间呈强正相关 (培养时间 :r=0 .873～0 .980 ,P<0 .0 5 ;铜离子浓度 :r=0 .90 0～ 0 .972 ,P<0 .0 5 )