波尔山羊FSHR5’端序列的SNP多态性及超排效果分析

为探索波尔山羊超排个体排卵数差异与个体遗传特性的相关性,对波尔山羊的FSHR基因(950 bp),分成四段分别设计引物进行PCR-SSCP多态性分析,结果发现一个SNP位点,导致5-’1出现两种基因型。序列分析显示该突变位点位于转录起始点上游-739bp处,属于远端转录启动调控区。统计分析表明AA型个体的平均排卵数与AB型个体的平均排卵数差异极显著(P<0.01),平均胚胎可利用率差异显著(P<0.05),这说明个体的超排潜力可能与其本身的FSHR基因的遗传特性有关。     

Postsurgical intra-incisional 5-fluorouracil in dogs with incompletely resected, extremity malignant spindle cell tumours: a pilot study

The safety and efficacy of intra‐incisional 5‐fluorouracil (5‐FU) in the management of incompletely resected malignant spindle cell tumours of extremities was evaluated in six dogs. After marginal surgery, the dogs underwent weekly intra‐incisional 5‐FU for a minimum of six cycles. Treatment was well tolerated by all dogs, with no systemic adverse effects and only one episode of local cutaneous hyperpigmentation, which completely and spontaneously resolved. Median follow‐up for all the dogs was 546 days (mean 619; range 297–1207). At the date of analysis, four dogs were still alive with no evidence of local recurrence, and two dogs had died as a result of their disease. The cause of death was development of distant metastases in one dog and tumour regrowth in the other. Despite the small sample size, this study documents that intra‐incisional 5‐FU chemotherapy is a safe and efficacious adjuvant treatment in the case of incompletely resected malignant spindle cell tumours in dogs and that long disease control can be achieved.     

猪泛素基因与PRRSV GP5基因融合表达质粒的构建及其免疫效果

构建了泛素（ubiquitin，Ub）基因与密码子优化的PRRSVGP5蛋白（optiGP5）基因融合表达质粒pIR—Ub—optiGP5。间接免疫荧光和Western—blot分析表明，重组质粒转染293T细胞后能够瞬时表达。将pIR—optiGP5和pIR—Ub—optiGP5两种质粒DNA肌肉注射免疫BALB／c小鼠后，分别检测小鼠血清中抗GP5抗体、T淋巴细胞的增殖能力以及CTL活性。结果显示，pIR—optiGP5质粒免疫组在二免后可以检测到荧光抗体，而pIR—Ub—optiGP5质粒免疫组一直未检测到抗GP5抗体，但该免疫组的T淋巴细胞增殖指数及针对GP5的特异性CTL反应数值明显高于pIR—optiGP5质粒免疫组。这表明泛素与GP5的融合表达可增强细胞免疫反应。     

肉鹅H5亚型禽流感疫苗免疫效果的研究

选用H 5亚型禽流感油乳剂灭活疫苗(H 5N 1,R e1株)对马岗鹅作首免日龄、免疫次数与免疫剂量的免疫试验。结果表明,雏鹅在7、14或21日龄作1次免疫均产生免疫应答,其中14与21日龄免疫组免疫后第5周抗体水平均达4 log2以上,抗体动态变化均值与峰值均高于7日龄免疫组2~3个滴度。用0.5、1和2 m l/只3个剂量对14日龄雏鹅的免疫试验中,0.5m l组抗体水平上升过程缓慢,免疫后4周抗体水平才达4 log2以上,1与2 m l组在免疫1周后上升速度较快,免疫后2周即达4 log2以上,抗体水平相近,但2 m l免疫组免疫后5~7周抗体水平均低于1 m l免疫组。说明接种剂量为1或2 m lH 5N 1油苗比接种0.5 m l的免疫效果好,尤以1 m l免疫剂量效果最好。雏鹅在7、14日龄或14、21日龄作2次免疫和7、14、21日龄作3次免疫,首免后抗体上升速度快,各检测点抗体水平均值相近,均在首免后3周达到4 log2以上,4~5周达6 log2以上,4~7周的抗体水平均高于14日龄1次免疫组,且提前2周达到4 log2以上,说明2次、3次免疫组的抗体水平高于1次免疫组,而2次与3次免疫组的抗体水平无明显差异。以上结果说明,在肉鹅生产的禽流感免疫中,选择以14日龄首免0.5 m l/只,21日龄2免接种1 m l/只可取得较为理想的免疫效果。     

A target-site mutation is present in a glyphosate-resistant Lolium rigidum population

Annual ryegrass (Lolium rigidum) is the only weed species to have evolved resistance to the broad‐spectrum herbicide glyphosate in Australia. A population that had failed to be controlled by glyphosate was collected from a vineyard in the Adelaide Hills region of South Australia. Dose–response experiments on this population (SLR 77) showed that it was glyphosate resistant, with an LD₅₀ that was 1.9–3.4 times higher than that of a susceptible population (VLR 1). The movement of radiolabelled glyphosate within SLR 77 plants showed that this population did not have the differential glyphosate translocation mechanism of resistance common to several other Australian glyphosate‐resistant populations. Subsequent analysis of shikimic acid accumulation within the plant after glyphosate treatment showed that this population accumulated significantly less shikimic acid than a susceptible population, but more than a glyphosate‐resistant population with the translocation mechanism, indicating the possible involvement of another mechanism of resistance. Sequencing of a portion of the SLR 77 5‐enolpyruvylshikimate‐3‐phosphate synthase gene was carried out and a mutation causing an amino acid change at position 106 from proline to threonine was identified. This mutation is likely to be responsible for glyphosate resistance in this population, as mutations in this position have been found to be responsible for glyphosate resistance in goosegrass (Eleusine indica) from Malaysia. This paper represents the first report of target‐site glyphosate resistance in L. rigidum and provides evidence that this species has at least two mechanisms of glyphosate resistance present in Australia.     

Studies of clomazone mode of action

Intact spinach chloroplasts were used to determine if clomazone, 5-OH clomazone, and/or 5-keto clomazone inhibited the chloroplastic isoprenoid pathway. When isopentenyl pyrophosphate was used as a precursor, neither clomazone nor the clomazone metabolites (5-OH clomazone and 5-keto clomazone) inhibited the formation of products separated by HPLC in the organic phase. However, when pyruvate, a substrate for the first committed step of the pathway, was used as a precursor, both 5-keto clomazone and fosmidomycin reduced the formation of a non-polar product and increased the formation of a polar product in the organic phase. Only 5-keto clomazone, not 5-OH clomazone or clomazone, inhibited the formation of an additional product other than fosmidomycin in the aqueous phase from pyruvate incorporation. In an in vitro assay, 5-keto clomazone inhibited DXP synthase, the enzyme catalyzing the first committed step of the chloroplastic isoprenoid pathway. Therefore, our studies show that neither clomazone nor 5-OH clomazone inhibits the chloroplastic isoprenoid pathway, only 5-keto clomazone does.     

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.     

用麦芽根调节水分对啤酒糟混贮品质的影响

将啤酒糟和麦芽根按不同比例袋装混贮,分别在30、60、90、120 d开袋取样进行感官评定,pH值、氨态氮、干物质、粗蛋白、中性洗涤纤维及有机酸含量分析。结果表明:混贮时间和水分含量均对混贮效果有显著影响;混贮饲料在混贮前60 d各指标变化范围较大,60 d后变化范围较小,并逐渐趋于稳定状态;混贮饲料以含水量50%的袋装混贮效果最好,其次是60%、40%,70%最差。     

表达禽流感病毒H5HA、H7HA基因DNA疫苗的免疫原性研究

为研究双顺反子DNA疫苗对禽流感病毒（AIV）的保护作用，将H5和H7亚型AIV的HA基因克隆到同一表达栽体上，构建了双顺反子HA基因表达质粒pCI—H5HA—H7HA。以此质粒肌注免疫4周龄SPF鸡，首次免疫后3周加强免疫，同时设pCI—H5HA和pCI—H7HA联合免疫组及空白对照组，每周采血用微量血凝抑制法检测HI抗体。加强免疫后3周分别以100LD50的高致病力禽流感病毒（HPAIV）A／Goose／Guangdong／1／96（H5N1）和A／FPv／Rostock／34〈（H7N1）进行致死性攻击。结果显示各免疫组均可刺激鸡体产生H5、H7特异性抗体，pCI—H5HA—H7HA诱导产生的抗体对H5N1和H7N1的攻毒保护分别为50%和10％，而pCI—H5HA和pCI—H7HA联合免疫的攻毒保护均为70％。表明双顺反子质粒可诱导鸡产生较好或一定的保护，但免疫效果不够理想，推测与DNA的摄取及其体内表达有关，可望通过调整DNA疫苗的多种因素提高免疫效果。     

鹅高致病性禽流感病理组织学观察

本文对发生H5N1高致病性禽流感禽场的鹅进行了病理学观察，证实此场鹅禽流感在剖检上以眼结膜潮红出血；心肌条纹状坏死，条带样出血；胰腺有白垩状或透明坏死点；胃肠出血等为特征。组织学观察以非化脓性脑炎，胰腺坏死，心肌坏死，坏死性脾炎为主要病变，揭示了鹅禽流感的病理学变化特征。