Genomic and Pathogenic Studies on a Glycoprotein E Variant Field Isolate of Bovine Herpesvirus 1

Glycoprotein E-negative (gE–) laboratory strains of bovine herpesvirus 1 (BHV-1) were recently introduced as novel marker vaccines, allowing serological discrimination between vaccinated and naturally infected animals on the basis of lack or presence of antibodies against gE epitopes. The applicability of this approach is based on the genetic stability of the gE. However, mutant field variants of BHV-1 with a variable response in anti-gE ELISA have been isolated. The molecular characterization of a gE variant field isolate (Salwa strain) is presented here. By comparing the gE nucleotide and amino acid sequences of the Salwa strain with those of the wild strain Jura, ten mutated bases were found in the gE strain of Salwa, six of which alter the amino acid sequence, leading to changes in five amino acids. Both strains caused respiratory disease in experimentally infected calves, but Salwa generated slightly milder signs. Both viruses were excreted in nasal and ocular discharges, and were reactivated by dexamethasone treatment. In conclusion, the rather close similarities observed in the gE gene structure and pathogenicity features of the gE mutant and of the wild strain of BHV-1 confirm the genetic stability of gE. The findings indicate that the Salwa isolate is virulent, but less virulent than wild strains. Our data support the use of gE-negative marker vaccines in eradication programmes.     

不同饲料蛋白质水平对吉富品系尼罗罗非鱼幼鱼生长和鱼体组成的影响

用3种不同蛋白含量(23.1%、37.6%和47.9%)的配合饲料投喂吉富品系尼罗罗非鱼幼鱼78d,结果显示,①随着饲料蛋白质含量从23.1%上升到47.9%,平均日增重从0.30g/d上升到0.74g/d,3组之间的日增重和体重有显著差异(P<0.05);饲料系数从1.53下降为0.98,3组之间的饲料系数有显著差异(P<0.05)。②不同的蛋白质水平对鱼体蛋白质和脂肪的含量有影响,各组之间差异显著(P<0.05),对灰分的影响差异不显著(P>0.05)。③蛋白含量为23.1%、37.6%和47.9%的配合饲料的成本分别为2.77元/kg、3.67元/kg、4.60元/kg。④初步认为蛋白含量为37.6%的配合饲料更适合吉富品系尼罗罗非鱼苗种培育的生产需要。     

产单核细胞李氏杆菌突变株的感染动力学研究

以BALB/c小鼠为实验动物模型,对actA基因和plcB基因双缺失的产单核细胞李氏杆菌（LM）yzuLM1-2菌株进行感染动力学研究。分别用减毒突变株yzuLM1-2和野生型菌株yzuLM4给BALB/c小鼠静脉注射或灌服,并对脏器中LM的分布动态进行测定。结果显示,在显著高于野生型菌株感染剂量的条件下注射或灌服接种后,减毒突变株组小鼠肝和脾中的LM数量迅速下降,被清除的速度显著快于野生型菌株组。溶血素（LLO）抗体的测定结果显示,减毒突变株菌能和野生型菌同样激发较高水平的抗体。小鼠免疫保护试验结果显示,减毒突变株免疫组对同种血清型LM的攻击具有较好的免疫保护力,可达100%。结果表明,yzuLM1-2突变株侵袭力下降,致病力降低,能激发较高水平的抗体产生,可以作为预防动物李氏杆菌病的候选疫苗菌株,并为用作运送外源保护性抗原的载体提供了材料。     

亚硝酸诱变选育黄霉素高产菌的研究

采用亚硝酸诱变方法对黄霉素产生菌进行诱变处理,利用菌株抗自身代谢物的特征,在添加高浓度黄霉素的分离培养基上选育获得优良高产突变株sg2-N2-2,比发酵菌株的发酵单位提高了7.2倍,而且变株的菌落形态与亲本菌株差异较大。     

Reclassification of an isolate of Guignardia citricarpa from New Zealand as Guignardia mangiferae by sequence analysis

Citrus black spot was recorded as present in New Zealand in international databases on the basis of one isolate (ICMP 8336) identified by morphological features as Guignardia citricarpa . This isolate was from a soft rot, not a typical symptom of citrus black spot, on a Seminole tangerine fruit grown in the northern region of New Zealand. Sequence analysis of the internal transcribed spacer (ITS) region (ITS1, 5·8S and ITS2) showed that this isolate was 99% identical to the ITS region of G. mangiferae , a closely related saprotroph. Despite climatic conditions being suitable for this disease, citrus black spot symptoms have never been seen on citrus fruits grown in New Zealand. Thus the absence of symptoms on citrus in New Zealand is probably because G. citricarpa is not present. On the basis of these results, the record of citrus black spot occurring in New Zealand should be re-examined.     

Study on Synergistic Effect of VA5 Immunopotentiator on Pigeon NDV Inactivated Vaccine

This study was designed to evaluate the synergistic effect of VA5 immunopotentiator on pigeon ND4416 inactivated vaccine.160 healthy pigeons were randomly divided into four groups,including ND4416 strain inactivated vaccine group (NDV group),ND4416 strain inactivated vaccine and immunopotentiator (VA5) mixed group (NDV+VA5 group),La Sota inactivated vaccine group (La Sota group) and normal saline as the control group (C group),to assess their vaccine efficacy against virulent pigeon NDV by serological analysis and animal testing.Pigeons sera were collected at different time points after immunization and measured the HI antibody titer of each group.The results showed that VA5 immunopotentiator significantly improved the serum antibody level of pigeons immunized with pigeon Newcastle disease inactivated vaccine (P<0.05).In addition,comparative test of spleen lymphocyte transformation was conducted at various time points after immunization.The results indicated that VA5 effectively stimulated the lymphocyte transformation of immune pigeon.Pigeons in each groups were challenged with ND4416 strain at the 30th,90th and 180th d after immunization.The results presented that the NDV+VA5 group had 100% protection rate and higher than La Sota group.The duration of immunization test showed that the antibody titer of NDV+VA5 group reached peak 11.20log₂ at the 21st d,remained 7.50log₂ at 180th d,and the protection rate remained 100% at 180th d.It indicated that VA5 immunopotentiator sustained the immune duration of pigeon NDV vaccine up to 180 d.Moreover,the in vitro detoxification test results suggested that VA5 immunopotentiator reduced the in vitro detoxification cycle of pigeons after challenge.Overall,this study suggested VA5 immunopotentiator could significantly improve the immune efficacy of pigeon Newcastle disease inactivated vaccine,which provided a basis for further enhancing the efficacy of Newcastle disease vaccine,as well increased experimental data for the application of immunopotentiators.     

猪伪狂犬病病毒双基因缺失突变株(HB-98株)安全性、稳定性和免疫原性测定

对以伪狂犬病病毒鄂A株为亲本毒株构建的TK和gG双基因缺失突变株（PrV HB-98株）的增殖能力、安全性、毒力稳定性和免疫原性进行了测定。结果表明，PrV HB-98株在BHK-21细胞上的增殖滴度为10^7.0 TCID50／0．1mL以上，与亲本毒株相当，但高于Bartha株；与PrV鄂A株相比，病毒量为10^7.0TCID50的PrV HB-98株不引起BALB／c小鼠的死亡，毒力也低于Bartha株；将PrV HB-98株在PK-15细胞连续培养25代和在猪体内上连续继代5次，各代次突变株TK基因和LacZ基因能被稳定扩增，未出现毒力回复现象．表明该毒株具有良好的遗传稳定性；以10^5.0、10^6.0、10^7.0TCID50等3个不同剂量的PrV HB-98株接种于妊娠50～60d母猪和1日龄仔猪，母猪均能正常产仔．仔猪也未出现任何临床症状，证明该毒株有较好的安全性。另外，以10^5.0TCID50的PrV HB-98株接种于妊娠50～60d母猪和1日龄仔猪，分别于接种后28d和20d，用10^7.0TCID50 PrV鄂A强毒进行攻击．结果免疫猪都能抵抗强毒的攻击．获得保护，表明该毒株具有很强的免疫原性。综合上述结果表明，PrV HB-98株可以作为候选毒株．用于伪狂犬病基因工程疫苗的研制。     

白灵菇15号杂交菌株选育研究

以白灵菇4号菌株和10号菌株为亲本，通过单孢杂交育种技术选育出白灵菇15号杂交新菌株。白灵菇15号杂交新菌株的子实体呈手掌形，柄较短，产量较高，平均生物学效率达48．26％。     

茶树新品系福选9号的观察研究

为观察鉴定茶树新品系福选9号在重庆市巴南区的适应性及表现,以福鼎大白茶为对照品种,于2002-2004年进行了品比试验。结果表明,福选9号表现出如下特性:(1)春芽萌动特早,较对照提早11～13d,春茶产量较高。(2)育芽能力强,生长势旺盛,秋梢休止期较对照晚5～7d,全年生育期较对照长16～20d,平均单产高20.5%。(3)抗逆性强,耐寒性和抗旱性与对照相当,抗病虫性稍强。(4)发芽整齐度高,适宜机械化作业。(5)制绿茶品质优良,适制性强。     

H9N2亚型禽流感病毒传播途径特性的比较及其表面基因序列分析

选择中国大陆最早分离的H9N2亚型禽流感病毒(avian influenza virus,AIV)A/Chicken/Guangdong/SS/94(H9N2)(缩写为SS株)和1998年大流行时期分离的H9N2亚型AIVA/Chicken/Shanghai/F/98(H9N2)(缩写为F株)为研究对象,对其在SPF鸡体内的复制能力和传播途径特性比较后发现,F株在4周龄SPF鸡气管中的复制能力高于SS株,F株可以经气溶胶传播途径传播,SS株不能经气溶胶传播途径传播;利用反转录-聚合酶链反应(RT-PCR)方法获取F株和SS株的HA和NA基因的cDNA,序列分析得知,F株和SS株的HA和NA基因的同源性分别是96.6%和98.1%;HA基因的裂解位点氨基酸序列都是PARSSR↓GL,但有5个氨基酸的差异,即166位N(F)→D(SS)、198位A(F)→V(SS)、217位V(F)→I(SS)、335位G(F)→R(SS)、504位L(F)→S(SS);2株病毒的NA基因在63~65位都存在氨基酸缺失,但在NA基因红细胞吸附位点的氨基酸序列不同,分别是IKKDSRSG(F)和IKEDLRSG(SS)。F株和SS株的传播特性差异是否与其表面基因序列有关,有待进一步研究。