球虫(菌)清缓释剂防治家兔球虫病和大肠杆菌病效果分析

目的寻求一种高效、优质药物防治家兔球虫病和大肠杆菌病。方法(1)60只健康兔分4组,第1、2组分别注射球虫清缓释剂、球菌清缓释剂,第3、4组分别用伯氨喹啉、地克珠利 氟哌酸拌料日常饲喂,于d 2、5、7、15、30口服球虫孢子化卵囊,隔10 d取粪查卵囊,观察保护率。(2)球虫病兔20只分为第1、3组,球虫病及大肠杆菌病兔20只分为第2、4组,分别用上述方法给药比较治愈率。(3)日常饲养兔150只分3组,分别为注射球虫清缓释剂、伯氨喹啉拌料日常饲喂、对照组,1、2、3个月后查卵囊,观察转阴率、OPG值、死亡率等。(4)幼兔100只分5组,前4组用方法(1)给药,第5组为对照组,比较增重率、饲料转化率、节省防治费率。结果(1)兔注射球虫清缓释剂、球菌清缓释剂后,在d 2~20时,攻击球虫,其保护率均为100%(3/3)。在d 30时,攻击球虫,其保护率均为66.7%(2/3);而口服伯氨喹啉,地克珠利 氟哌酸防治的兔,在d 2~5时攻击球虫,保护率为0,在d 10时,其保护率均为33.3%(1/3),在d 20~30时,其保护率均为66.7%(2/3)。(2)球虫病和大肠杆菌病兔(1、2组)注射球虫清缓释剂、球菌清缓释剂后d 5~60,球虫病、大肠杆菌病的治愈率均为100%;口服伯氨喹啉、地克珠利 氟哌酸(3、4组)后d 5,治愈率均为0,d 20~60时,治愈率为70.0%~90.0%。(3)球虫清缓释剂组在1~2个月时,球虫卵囊减少率及转阴率均为100%,在3个月时,球虫卵囊减少率为98.9%,转阴率为94.0%,存活率为100%。伯氨喹啉组1~3个月,球虫卵囊减少率及转阴率分别为64.4%~89.8%及33.0%~91.2%,死亡率为22.0%~28.0%,存活率为72.0%~78.0%。对照组1~3个月,球虫卵囊增加78.0%~126.0%,死亡率为46.0%~62.0%,存活率为38.0%~54.0%。(4)注射球虫(菌)清缓释剂比口服伯氨喹啉、地克珠利 氟哌酸及对照组,兔的增重率分别提高71.7%、76.1%、374.6%和69.8%、74.1%、369.3%,节省防治费75%。应用球虫(菌)清缓释剂防治球虫病、大肠杆菌病兔10.5万只,获得经济效益108.67万元(包括增重效益、挽回死亡兔损失、减少饲料费和节省防治费四项)。结论球虫(菌)清缓释剂能安全、高效、经济、方便地控制家兔球虫病和大肠杆菌病的发生,保护幼兔安全度过球虫病和大肠杆菌病高危期。     

家禽病毒性免疫抑制病的分析与对策

免疫抑制引起免疫失败及对病原的易感性增加，是目前乃至今后威胁家禽健康的最危险的潜在因素之一。作者阐述与讨论了几种目前常见的家禽病毒性免疫抑制病的概念、免疫抑制机理及防制措施，以此引起同行对该病的进一步重视。     

鸡新城疫病毒地方株的分离鉴定及其遗传变异分析

从河北省保定地区发病鸡群中分离到2个具有血凝性的病毒株分别命名为HBA和HBB,其血凝作用可被NDV阳性血清抑制,而不能被AIV(H9亚型)、EDS76阳性血清抑制,表明2个分离株均为新城疫病毒。通过测定MDT、ICPI和IVPI等方法鉴定分离株的毒力符合强毒标准。利用RT-PCR技术成功扩增了2株分离株的F基因片段(约540bp),通过测序及遗传变异分析表明两毒株之间的核苷酸同源性为87.7%,氨基酸同源性为91.6%,结合系统发育进化树分析表明HBA为基因Ⅵ型,HBB为基因Ⅶ型。     

新城疫病毒F、NP、M和HN基因在昆虫细胞内的共表达

为构建杆状病毒转移载体，通过PCR的方法将F48E9株新城疫病毒F基因上的StuⅠ位点和NP基因上的XbaⅠ位点进行突变，扩增出全长的F、NP以及F48E9株新城疫病毒M和HN基因片段，将其克隆到pMD18-T载体，再将F、NP、M和HN基因依次亚克隆到杆状病毒转移载体pAeAB4上，F基因和M基因均在p10启动子的操控之下，NP基因和HN基因构建到两个polyhedrin启动子下游，构建重组杆状病毒转移载体pAeAB4-F-NP-M-HN。pAeAB4-F-NP-M-HN与线性化的杆状病毒DNA共转染昆虫细胞Sf9，获得重组杆状病毒rBae-F-NP-M-HN。重组杆状病毒感染Sf9细胞，72h后收集细胞和培养上清。Western blot分析显示：M、NP、F和HN蛋白在培养上清中得到了共表达，大小与预期结果一致；感染细胞内只检测到了HN蛋白的表达。这表明M、NP、F和HN蛋白在昆虫细胞内共表达可以自我装配成病毒样颗粒，并且以出芽的方式释放到培养基中。该重组杆状病毒的获得为研究新城疫病毒各结构蛋白之间的相互作用和确定病毒粒子出芽的驱动力等方面奠定了基础。     

表达鸡马立克氏病病毒gB基因重组鸡痘病毒的遗传稳定性及生物安全性评价

为了评价表达鸡马立克氏病病毒gB基因重组鸡痘病毒(rFPV-gB/R)的遗传稳定性,我们将纯化后的重组病毒在CEF单层上连续传30代,引起细胞病变的速度和形态均未发生明显变化;覆盖含X-Gal的琼脂引起的空斑均为蓝色;间接免疫荧光实验证明rFPV-gB/R中的gB基因始终能稳定表达;序列测定结果表明,重组病毒在细胞上连续传30代、在SPF鸡上连续传5代后,gB基因序列没有发生任何变化;以0、10、20、30代重组病毒制成冻干疫苗进行的实验室免疫效力实验表明,细胞连续传代后rFPV-gB/R仍然保持了原有的免疫原性。可见重组鸡痘病毒gB基因的结构和免疫原性都是高度稳定的。为了评价rFPV-gB/R的生物安全性,我们将rFPV-gB/R通过SPF鸡连续传5代,检测病毒在鸡体的存在部位及其消长、生长繁殖性能和毒力变化;将rFPV-gB/R免疫鸡与未免疫鸡同笼饲养,攻击FPV-102E6强毒,以检测rFPV-gB/R感染鸡的接触传染性。结果显示,rFPV-gB/R在鸡体的存在时间大约为7d,在体内仅存在于接种部位;鸡体传代后痘病毒毒力有一定程度下降,gB基因核苷酸序列未发生任何变化;同居未免疫SPF鸡在痘病毒强毒攻击后全部发痘,可见重组病毒免疫鸡没有接触传染性,能在鸡体内稳定地传代,rFPV-gB/R具有高度的生物安全性。     

头孢噻呋对实验性感染鸡白痢沙门氏菌病的药效学研究

研究了头孢噻呋对实验性感染鸡白痢沙门氏菌的防治效果。结果显示，头孢噻呋体内疗效良好，治疗组与预防组的治愈率、有效率、相对增重率均不同程度高于感染对照组（P〈0.05或P〈0.01）；头孢噻呋的治疗效果高剂量组与中剂量组无显著差异（P〉0．05），高剂量组与低剂量组差异显著（P〈0．05）；头孢噻呋的中剂量组与头孢曲松、氨苄西林组的疗效相比均差异显著（P〈0．05或P〈0．01）。实验结果表明，头孢噻呋按0．1mg／只的剂量肌肉注射，每日一次，可对鸡白痢沙门氏菌病起到有效的防治效果。     

Influences of coculture with bone marrow mesenchymal stem cells on dopaminergic neurons in brain slice treated with 1-methyl-4-phenylpytidinium

AIM: To study the protective effects of bone marrow-derived mesenchymal stem cells on damaged dopaminergic neurons induced by 1-methyl-4-phenylpytidinium(MPP+).METHODS: The parkinson disease(PD) models were established in newborn rats. Bone marrow mesenchymal stem cells(MSCs) obtained from adult bone marrow were cultured, isolated and purified. MSCs were co-cultured with brain slice and the immunohistochemical technique, electron microscopy, propidium iodide staining were used to observe the changes of neurons. RESULTS: In the MPP+ treatment group, the neurites grew slowly and sparsely, dead cells were found in all regions. In the co-culture group, the neuritis grew densely, only a few cells were dead, the number of tyrosine hydroxylase(TH)-stained neurons increased and the structure of organellae was normal. CONCLUSION: MSCs may protect dopaminergic neurons against damage induced by MPP+. These results provide some data for cell transplantation therapy to Parkinsons disease.     

Aloe ring spot,a new disease of aloe caused by <Emphasis Type="Italic">Haematonectria haematococca</Emphasis> (Berk. &; Broome) Samuels &; Nirenberg (anamorph: <Emphasis Type="Italic">Fusarium</Emphasis> sp.)

A ring spot disease of Aloe vera was found on leaves of potted seedlings of Aloe vera in Hachijojima and Chichijima Islands, Tokyo. From tissue of ring spot lesions, a fungus producing Fusarium-type conidia was consistently isolated. After 1 month, reddish perithecia of nectriaceous fungus had formed on the colonies of this isolate on PDA. These nectriaceous and Fusarium fungi were identified as Haematonectria haematococca and Fusarium sp., respectively. From a single ascospore isolation, the former was confirmed to be the teleomorph of the Fusarium sp. Typical ring spot lesions were reproduced by artificial inoculations using single ascospore and single conidium isolates. Inoculations of five species of genus Aloe revealed that they were highly susceptible except for A. arborescens. This is the first report of a disease on Aloe caused by H. haematococca (anamorph: Fusarium sp.) in Japan, and it was named aloe ring spot.     

Lesion-based analysis of leaf blast suppression in mixture of rice cultivar and a resistant near-isogenic line

Leaf blast suppression in multilines was evaluated based on the number of susceptible lesions observed in a pure stand of susceptible rice cultivar Sasanishiki, and in 1 : 1 and 1 : 3 mixtures of Sasanishiki and a resistant near-isogenic line, Sasanishiki BL4 or BL7, from 1998 to 2001. The number of lesions first observed in fields in the 1 : 1 and 1 : 3 mixtures were close to theoretical numbers calculated using the number of lesions observed in the pure stands and the ratios of the susceptible Sasanishiki in the mixtures. The ratio of the number of lesions in the 1 : 1 and 1 : 3 mixtures to the number in the pure stand was 0.29 and 0.09, respectively. The relationship between these ratios and the ratios of susceptible Sasanishiki in mixtures was defined in an equation to estimate the degree of leaf blast suppression. Validation studies for the ratios of the number of lesions in the 1 : 1 and 1 : 3 mixtures to the number in the pure stand were conducted in two different locations and showed that the ratios are almost acceptable. The calculated autoinfection to alloinfection ratio was 1.3 and 1.4 in the 1 : 1 and 1 : 3 mixtures, respectively, suggesting that the calculated ratio will affect the degree of leaf blast suppression. Thus, predictors were obtained to estimate leaf blast suppression for effective blast control in multilines.     

Occurrence of <Emphasis Type="Italic">Phaeoacremonium</Emphasis> spp. and <Emphasis Type="Italic">Phaeomoniella chlamydospora</Emphasis> in grape propagation materials and young grapevines

During a 6-year study, grapevine propagation materials and young grapevines were analysed to evaluate the presence of internal wood discolouration and the occurrence of fungal species involved in Petri disease. The intensity of wood discolouration increased with the ageing of the plants. The maximum incidence of dark streaks was observed in the rootstock while necrosis originating from buds or nodes were notably present in the trunk and cordon of older vines. In contrast, the highest levels of brown-red halo symptoms, defined as discoloured areas around the pith, were recorded in the early growth stages. Phaeoacremonium spp. and Phaeomoniella chlamydospora were usually isolated from the rooted-grafts and the 3-year old plants, respectively. The number of infected grapevines increased with age. Most of the P. chlamydospora strains were isolated from dark streaks or dots, while Phaeoacremonium spp. were detected in brown-red halo symptoms and other symptomatic or asymptomatic wood. The greatest incidence of the two fungal taxa was recorded in the lower parts of the grapevine, including the roots and rootstock.