液相芯片技术同时检测六种猪繁殖障碍病毒的方法研究

在GenBank中收集猪细小病毒(PPV)结构蛋白VP2基因、猪圆环病毒2型(PCV2)的ORF2序列、猪伪狂犬病毒(PRV)的gB基因、猪繁殖与呼吸综合征病毒(PRRSV)的NSP2基因、猪瘟病毒(CSFV)的E2基因和乙型脑炎病毒(JEV)的M基因,利用Primer5.0等分子生物学软件对收集的序列分析比较,筛选出上述基因的特异保守序列并设计引物和探针,将设计好的探针与相应的微球偶联,优化条件,建立检测方法。结果表明:建立的检测PPV、PCV2、PRV、PRRSV、CSFV、JEV6种病毒核酸的多重液相芯片技术具有较好的特异性、敏感性和稳定性,对PPV、PCV2、PRV、PRRSV、CSFV、JEV6种病毒核酸的最低检出量分别为8.50×10~2copies/μL、2.87×10~2copies/μL、2.07×10~2copies/μL、2.61×10~2copies/μL、2.34×10~2copies/μL、2.05×10~2copies/μL,与相应病毒PCR/RT-PCR检测方法相比,敏感性提高了100~1000倍;对临床388份样品同步进行荧光PCR与液相芯片检测,结果99.87%相符。本方法为液相芯片技术在动物多病毒快速高通量检测、鉴别诊断等应用方面奠定了基础。     

Polymorphism of <Emphasis Type="Italic">Got2</Emphasis> DNA sequences sheds light on <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss. intraspecies divergence and origin of <Emphasis Type="Italic">Triticum aestivum</Emphasis> L.

DNA sequences of nuclear gene Got2 was studied in 60 accessions of Aegilops tauschii, 29 of subsp. tauschii and 31 of subsp. strangulata. It was found that Got2 allozyme polymorphism in Ae. tauschii is due to a single, unique, mutation which led to replacement of glutamic acid by isoleucine in residue 256 of the enzyme molecule, encoded by Got2. As revealed by Got2 DNA sequences variation, initially in its history Ae. tauschii was presented by subsp. strangulata, and among phylogenetic lineages of subsp. strangulata, the lineage “t-9¹s” (TauL3) is the most ancient, a relict one. Subspecies tauschii is relatively “young”. Initially it was presented by the lineage marked by combination of allozyme alleles Got2 ¹⁰⁵ and Acph1 ¹⁰⁰. In the past it inhabited the Continental area from Caucasia to Pakistan, but later on it was forced out by newly originated, now—a major lineage of subsp. tauschii, marked by Got2 ¹⁰⁰. This lineage extended the Continental area of the species up to Kirgizstan, but actually failed to penetrate into pre-Caspian area, occupied by subsp. strangulata. These results essentially differ from those obtained previously, using chloroplast DNA (cpDNA) sequences polymorphism. As revealed by cpDNA, the major, “usual”, subsp. strangulata (TauL2) is “younger” than subsp. tauschii, which resided on phylogenetic tree between relict lineage “t-9¹s”of subsp. strangulata—and major subsp. strangulata. But both cpDNA and Got2 DNA sequences indicate that the level of genetic variation in subsp. tauschii is much lower than in subsp. strangulata. According to Got2 DNA sequences variation, it was Ae. tauschii subsp. strangulata lineage “k-109″ which donated genome D to Triticum aestivum L. This lineage includes accessions: k-109 from South-Eastern Precaspian Azerbaijan; KU-2105, KU-2159 from Western Precaspian Iran; KU-2080 from Eastern Precaspian Iran.     

高山杜鹃成都平原越夏生长研究

为加强对高山杜鹃的引种栽培,筛选出适合成都平原栽植的高山杜鹃品种,对引进的11个品种高山杜鹃的整株生长指标（冠幅、株高、地径、分蘖能力）、枝条生长指标（分枝数、最长分枝长度、单枝粗度、最长枝叶片数）、叶片生长指标（叶长、叶宽、叶片厚度）、夏稍长度以及热害指数进行评估。结果表明,引进的11个品种高山杜鹃均能在一定程度上适应成都平原越夏生长,其中‘王妃’的综合表现最好,尤其是其冠幅、株高、地径、最长分枝长度、单枝粗度、叶长、叶宽,在引种2个月后的增幅分别达到了131.43%、155.07%、53.79%、103.65%、26.66%、21.51%、27.34%;‘喜悦’的热害指数最低,几乎不受害。此外,‘粉蜜’、‘红溢’也表现不俗。因此,引进高山杜鹃品种‘王妃’、‘粉蜜’、‘红溢’、‘喜悦’适应成都平原的越夏生长,可进一步推广栽植。     

环境温度、饲料、性别对肉鸡氨基酸表观消化率和氮、能代谢的影响

利用72只32日龄京白904肉仔鸡公母各半研究环境温度简称温度、饲料种类和鸡的性别对氨基酸表观消化率AAAA、氮能代谢的影响。试验采用3×2×2析因设计。试验鸡在8℃、24℃、30～32℃下处理14天,43日龄时,采用改进的真代谢能TME法测定3种温度下肉用公母鸡对玉米和豆粕的AAAA、代谢能及与氮代谢有关的指标。试验结果如下:饲料种类极显著地影响AAAAP<0.01。温度和性别对AAAA测值无显著影响P>0.05。内源干物质EDM排量不受温度的影响P>0.05。高温组表观代谢能AME显著高于低温组P<0.05,而常温组与高温组及低温组间差异不显著P>0.05。性别间AME无显著差异P>0.05。温度及性别对氮校正表观代谢能AMEn无显著影响P>0.05。尿酸FUA受温度影响极显著P<0.01。氮沉积率NR不受温度的影响P>0.05。氮平衡值NB受温度影响极显著P<0.01。     

刚地弓形虫亲环蛋白基因TgCyP真核表达质粒的构建及其在Hela细胞内的表达

亲环蛋白（cyclophilin,CyP）是一类广泛存在于原核和真核生物体内的胞溶性蛋白,是刚地弓形虫（Toxoplasmngondii）速殖子的主要成分,能够诱导产生IL-12和IFN-γ,在控制弓形虫急性感染过程中起重要作用。本研究根据GenBank发表的TgCyP基因序列,设计并合成一对包含BamHI和EcoRI酶切位点的引物,以cDNA为模板,应用PCR技术扩增TgCyP基因。PCR产物连接到pMD18-T克隆载体。用限制性内切酶BamHI和EcoRI双酶切克隆载体,将TgCyP目的基因克隆到载体pVAXI,构建真核表达质粒pVAXl-TgCyP。将真核表达质粒转染Hela细胞,利用问接免疫荧光检测其在Hela细胞内的表达情况。结果表明扩增的TgCyP基因与GenBank上相应基因序列（U04633．1）的一致性达100％,构建的真核表达质粒pVAX1-TgCyP能在转染的Hela细胞中表达,其表达产物与刚地弓形虫阳性血清具有免疫反应性。本研究表明Tg-CyP有望作为弓形虫疫苗的候选抗原,将为进一步研究该质粒的动物免疫试验奠定基础。     

Management of pine wilt disease in Korea through preventative silvicultural control

Pine wilt disease (PWD) represents a major threat to forest ecosystems worldwide. Although PWD is now better understood, effective control measures for this disease have still not been devised. Here, we report several years of field studies on preventative silvicultural control of PWD. Silvicultural control through preventative clear-cutting and the manual removal of logs was implemented between 2005 and 2009 in 16 Korean districts that had newly PWD-infected stands. Preventative clear-cutting of neighboring asymptomatic pine trees (within a 10-50-m radius of wilt trees) and the removal of felled logs or branches suppressed spread of PWD. Occurrences of PWD wilt pines in districts (city or county) subjected to this silvicultural control method were significantly reduced compared with those in districts using conventional controls (physical or chemical treatment of wilt pine trees). Through silvicultural control, PWD was successfully suppressed in 11 of 16 districts investigated. In contrast, successful control was achieved in only 1 among 18 districts subjected to conventional control. Our results will be of considerable interest to those engaged in the very difficult battle against the global spread of PWD.     

Influence of green tea polyphenol in rats with arginine-induced renal failure

To determine whether green tea polyphenol ameliorates the pathological conditions induced by excessive dietary arginine, green tea polyphenol was administered to rats at a daily dose of 50 or 100 mg/kg body weight for 30 days with a 2% w/w arginine diet. In arginine-fed control rats, urinary and/or serum levels of guanidino compounds, nitric oxide (NO), urea, protein, and glucose increased significantly, while the renal activities of the oxygen species-scavenging enzymes superoxide dismutase (SOD) and catalase decreased, compared with casein-fed rats. However, rats given green tea polyphenol showed significant and dose-dependent decreases in serum levels of creatinine (Cr) and urea nitrogen and urinary excretion of Cr, and they exerted a slight reduction of nitrite plus nitrate, indicating that green tea polyphenol reduced the production of uremic toxins and NO. In addition, in arginine-fed rats the urinary urea, protein, and glucose level increases were reversed by the administration of green tea polyphenol. Moreover, in rats given green tea polyphenol the SOD and catalase activities suppressed by excessive arginine administration increased dose-dependently, implying the biological defense system was augmented as a result of free radical scavenging. These results suggest that green tea polyphenol would ameliorate renal failure induced by excessive dietary arginine by decreasing uremic toxin, and NO production and increasing radical-scavenging enzyme activity.     

水牛MFSD2A基因多态性及其与产奶性状的关联分析

【目的】研究水牛主要促进因子超家族成员2a(major facilitator superfamily domain containing 2a, MFSD2A)基因多态性,并筛选与水牛产奶性状相关联的SNP。【方法】采集383头健康水牛血液DNA,利用PCR扩增和Sequenom MassARRAY飞行时间质谱技术对SNP位点进行筛选及基因型检测,并进行遗传多样性、连锁不平衡(LD)和单倍型分析,以及对不同基因型和单倍型与水牛产奶性状进行关联分析。【结果】在水牛MFSD2A基因中检出7个SNPs位点,多态信息含量(PIC)均在0.25～0.40之间,属于中度多态,除g.8290 T>C位点外,其余位点均处于Hardy-Weinberg平衡状态(P>0.05);关联分析结果表明,这7个SNPs位点与305 d产奶量均显著关联(P<0.05),且突变杂合子基因型的305 d产奶量是3种基因型中最低的,g.568 C>G和g.701 A>G位点与乳脂率显著关联(P<0.05),乳脂率从大到小依次为突变纯合子>野生纯合子>突变杂合子,g.568...     

柘林湾网箱养殖海域溶解氧分布及其影响因素

根据１９９８年７月在南海东北部柘林湾的调查数据，探讨网箱养殖海域溶解氧的分布及其与其他环境因子间的关系，结果显示夏季海水溶解氧浓度为３.03-7.86mg/L( 平均５．３０）,退潮时（平均５.71)高于涨潮时（平均４.65)涨潮时网箱区内贫氧（ ＜０．mg/L).多元逐步回归分析表明，对溶解氧浓度有重要影响的水质因子为pH和水温，其次为盐度，ＣＯＤ和营养盐，它们的变化与ＤＯ的变化有较好的相关性，区域比较表明，海水溶解氧浓度与观测僧置到网箱区的距离呈正相关（Ｐ＜０．０２），与底质硫物，ＰＯ４－Ｐ含量呈负相关（Ｐ＜０．０５）,溶解氧浓度偏低与养殖环境的污染有直接关系。     

Pharmacokinetics after administration of an injectable experimental long-acting parenteral formulation of doxycycline hyclate in goats

OBJECTIVE: To determine the pharmacokinetics after SC administration of an experimental, long-acting parenteral formulation of doxycycline hyclate in a poloxamer-based matrix and after IV and IM administration of an aqueous formulation of doxycycline hyclate in goats. ANIMALS: 30 clinically normal adult goats. PROCEDURES: Goats were allocated to 3 groups (10 goats/group). One group of goats received doxycycline hyclate (10 mg/kg) IM, a second group received the same dosage of doxycycline hyclate IV, and the third group received the long-acting parenteral formulation of doxycycline hyclate SC. Serum concentrations of doxycycline were determined before and at various intervals after administration. RESULTS: The long-acting parenteral formulation of doxycycline hyclate had the greatest bioavailability (545%); mean +/- SD maximum serum concentration was 2.4 +/- 0.95 microg/mL, peak time to maximum concentration was 19.23 +/- 2.03 hours, and elimination half-life was 40.92 +/- 4.25 hours. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the long-acting parenteral formulation of doxycycline hyclate distributed quickly and widely throughout the body after a single dose administered SC, and there was a prolonged half-life. Bioavailability of the longacting parenteral formulation of doxycycline hyclate after SC administration was excellent, compared with bioavailability after IV and IM administration of an aqueous formulation of doxycycline hyclate. Although no local tissue irritation and adverse effects were detected, clinical assessment of drug-residues and toxicologic evaluations are warranted before this long-acting parenteral formulation of doxycycline hyclate can be considered for use in goats with bacterial infections.