禽腺病毒血清4型贵州株全基因序列分析

为了解禽腺病毒血清4型(FAdV-4)地方流行毒株的分子进化情况,基于实验室分离的2株FAdV-4贵州株GZ-BJ株和GZ-QL株,分别对2株FAdV-4毒株进行PCR分段扩增,扩增产物克隆至载体,提取质粒进行PCR和双酶切鉴定后筛选出重组质粒进行测序,将测序结果依次拼接得到病毒的全基因组,获得FAdV-4贵州株的全基因序列,并对其进行序列和遗传进化分析。结果显示,通过PCR分段扩增成功获得了2株FAdV-4贵州株(GZ-BJ株和GZ-QL株)的全基因序列,长度分别为43352、43723 bp,FAdV-4 GZ-BJ株全基因序列长度比FAdV-4 GZ-QL株短371 bp,少6个ORF(22K、putative 9.1 ku、u-exon、ORF17、ORF28、ORF42),二者的氨基酸同源性为57.1%。2株FAdV-4贵州株同国内外不同地区FAdV-4毒株核苷酸同源性在88.7%~100%,与FAdV-4经典毒株ON1比对,2株FAdV-4贵州株和国内FAdV-4分离株均缺失ORF19、ORF27、ORF30。系统进化树分析显示,2株FAdV-4贵州株GZ-BJ株和GZ-QL株仍属于Ⅰ群C种FAdV。研究结果表明,2株贵州株FAdV-4 GZ-BJ株和FAdV-4 GZ-QL株较国内外FAdV-4毒株均存在进化与突变,且FAdV-4 GZ-BJ株变化较大,但尚未改变其血清型,这为探索FAdV-4致病机理的分子机制研究提供依据。     

２，４-二叔丁基苯酚对啤酒花幼苗生长与光合特性的影响

前期对不同植龄啤酒花根系分泌物及根际土壤的化学物质进行了分离鉴定,结果发现有酚类物质的存在且２,４-二叔丁基苯酚含量较高,为了进一步证实啤酒花是否存在自毒作用,采用田间小区栽培试验,研究了不同用量［０（ＣＫ,Ａ０）,７．５（Ａ１）,１５．０（Ａ２）和２２．５ｍｍｏｌ／ｍ２（Ａ３）］２,４-二叔丁基苯酚处理对啤酒花幼苗生长及光合特性的影响。结果表明,２,４-二叔丁基苯酚用量Ａ１ 和Ａ２ 处理,啤酒花生长好,地上部、地下部、总生物量及日均光合速率均比对照高,且达到显著差异（犘＜０．０５）;２,４-二叔丁基苯酚用量Ａ３ 处理,啤酒花地上部、地下部和总生物量及光合特性均较对照降低,差异达到显著水平（犘＜０．０５）。啤酒花幼苗叶片光合速率日变化呈双峰型,最高峰出现在上午９：００左右,次高峰出现在中午１５：００左右,午间有明显的“午休“现象,与对照（Ａ０）相比,Ａ１、Ａ２、Ａ３ 三个处理的次高峰出现的时间均有推迟,且Ａ１ 和Ａ２ 处理的幼苗午休”现象不太明显,气孔因素是导致其“午休”的主要原因。     

猪粪中除臭微生物的筛选和鉴定

用感官法初筛出20株菌株,再通过定性复筛试验,最后从猪粪便中共分离出5株除臭微生物,都具有很强的除臭效果,其中假单孢菌属(ZY-1)氨气的去除率高达47.70%,巴氏芽孢杆菌(WJ-2)硫化氢的去除率高达62.5%,显示出这类细菌在猪场粪便的处理过程中具有良好的应用前景。     

黄河源区“黑土滩”退化草地CO2排放研究

采用静态箱-气相色谱法,对黄河源区“黑土滩”退化草地CO2释放特征的研究结果表明：“黑土滩”3种不同处理：“黑土滩”植被-土壤系统呼吸（BC）,土壤呼吸（BJ）和土壤微生物呼吸（BL）的CO2释放速率日变化具有相似性,均呈现出明显的单峰型特点,且白天大于夜间;CO2释放速率季节动态变化明显,且3者的变化趋势基本一致;“黑土滩”植被-土壤系统呼吸与地表温度及5cm地温呈极显著的正相关性（P〈0．01）;而土壤微生物呼吸与0～10cm土壤水分含量相关显著（P〈0．05）。     

猪圆环病毒2型核酸疫苗的小鼠免疫保护试验

为了研究猪圆环病毒2型(PCV2)核酸疫苗在小鼠攻毒试验中的免疫保护效果,以PCV-2 GXWZ-1株为模板,扩增出ORF2基因及其截短基因8个片段(A(ORF2)、B(51-100aa)、C(101-150aa)、D(181-235aa)、E(151-200aa)、F(51-150aa)、G(101-235aa)、H(51-235aa)),将其插入到pcDNA3.0载体中,构建出真核表达质粒,并将其转染至PK-15细胞,用间接免疫荧光试验检测其瞬时表达情况。将试验小鼠随机分成9组,其中免疫组7组,阴阳性对照各1组,将纯化的真核表达质粒对小鼠进行组合免疫;二免后,用经处理过的PCV-2 GXWZ-2株阳性病料悬液腹腔注射免疫组和非免疫对照组小鼠,阴性对照组用生理盐水腹腔注射;其后进行体重记录、病理切片制作及PCR检测。结果表明:共有6个真核表达质粒成功在PK-15细胞中表达。在攻毒后的3周内,阳性对照组小鼠PCR诊断均为阳性;免疫组中,部分组小鼠在攻毒后第1周或在第2周为阳性,到第3周时各免疫组小鼠全部为阴性;阴性对照组始终为阴性。免疫组在病理保护学方面明显优于非免疫对照组,非免疫对照组的体重增长速率略低于免疫组和阴性对照组。由此可见猪圆环病毒2型核酸疫苗在小鼠攻毒试验中有明显的保护作用。     

猪传染性胸膜肺炎放线杆菌和猪圆环病毒2型二重实时荧光定量PCR检测方法的建立

以胸膜肺炎放线杆菌(App)apx ⅣA基因和猪圆环病毒2型(PCV-2)ORF2基因为靶基因,分别设计特异性引物和探针,建立App和PCV-2的二重荧光PCR方法。App引物扩增的目的片段大小为132bp,PCV-2为169bp。建立的方法可在同一反应体系中同时对App和PCV-2进行定量检测鉴别,其敏感性分别为1×101拷贝和1×100拷贝,对其他猪病病原核酸的检测为阴性。利用该方法对10份人工感染临床样品进行检测试验,其结果符合率达100%,说明该方法可用于对临床样品中App和PCV-2的检测。     

Closed system anaesthesia in dogs using liquid sevoflurane injection; evaluation of the square-root-of-time model and the influence of CO2 absorbent

Objective To determine whether predictable alveolar concentrations of sevoflurane are reliably produced in dogs when liquid sevoflurane is injected into closed circuit breathing systems, as calculated by Lowe's square‐root‐of‐time anaesthetic uptake model, and to confirm the validity of the model using soda lime and calcium hydroxide lime. Study design Prospective clinical study. Animals Eleven healthy dogs with a mean body mass of 34 ± 9 kg scheduled for pelvic limb orthopaedic surgery. Materials and methods Following pre‐anaesthetic medication, anaesthesia was induced with propofol and maintained with sevoflurane in a closed circle system. Epidural anaesthesia was performed with morphine and bupivacaine. Liquid sevoflurane was injected into the circuit by syringe, using dosages and time intervals derived from Lowe's square‐root‐of‐time anaesthetic uptake model. The target alveolar concentration chosen was 1.1 × MAC (2.6% end‐tidal sevoflurane). Either soda lime (group S; n = 6) or calcium hydroxide lime (Amsorb; group A; n = 5) were used for CO₂ absorption. Sevoflurane concentration and the respiratory gas composition were measured with an infrared gas analyser. Results End‐tidal sevoflurane concentrations were close to the predicted value of 2.6% at 9 minutes (2.53 ± 0.1% group S; 2.60 ± 0.26% group A) and 16 minutes (2.55 ± 0.30 group S; 2.52 ± 0.28% group A) but declined thereafter to reach 50% (group S) and 64% (group A) of the predicted value at 121 minutes. There was a constant trend towards higher end‐tidal sevoflurane concentrations in group A but the difference was not statistically significant. Conclusions The square‐root‐of‐time model leads to significantly lower alveolar concentrations than expected, suggesting that the rate of sevoflurane uptake in dogs declines less rapidly than predicted. The use of Amsorb tends to reduce the deviation from predicted concentrations. Clinical relevance The model used in this study provided only an approximate guide to the volume of liquid sevoflurane required. Consequently, the definitive dose schedule must be based on measured anaesthetic concentrations and clinical monitoring.     

First report of porcine circovirus type 2 infections in Cuba

To obtain information about the porcine circovirus type 2 (PCV2) infection status of pigs in Cuba and the probable association of PCV2 with other porcine viruses, tissue samples collected from ill pigs were evaluated using polymerase chain reaction (PCR). The PCR analysis showed that 67.7% of the samples (23/34) from seven swine herds of six different geographic regions were detected to be positive for PCV2. Ten of the 23 PCV2 positive samples (43.5%) shown a concurrent infection with porcine parvovirus (PPV) and 17 of 23 PCV2 positive samples (73.9%) exhibited a concomitant infection with classical swine fever virus (CSFV). This study is the first report of PCV2 infecting pigs with different clinical conditions in Cuban swine herds and provides evidence of PCV2 co-infection with PPV and CSFV in the field.     

南阳牛IGF2基因第2外显子多态性及其与生长发育相关性研究

以126头纯种南阳黄牛为研究材料，利用PCR-RFLPs技术对胰岛素样生长因子2（IGF2）第2外显子的多态性及其与生长发育的相关性进行了分析。结果表明：南阳牛在该位点的遗传多态性丰富。检测到2种等位基因A和B，频率分别为0．6865和0．3135，A等位基因在群体中是优势等位基因；BB型个体的6、12、18、24月龄体斜长和胸围，12、18和24月龄的坐骨端宽，初生、6月龄和24月龄体重等体征性状显著地优于AA型；6月龄体斜长、12月龄的胸围、18月龄和24月龄坐骨端宽也显著优于杂合型。初步推断JGF2基因是非常有价值的辅助选择标记，可为南阳牛肉用型方向选择提供理论依据。