Viral respiratory disease of the horse.

The diagnosis of any viral respiratory disease relies on laboratory procedures to isolate the virus and demonstrate a significant rise in serum antibody titers. To isolate viruses from the upper respiratory tract, it is imperative that nasopharyngeal swabs are obtained from animals in the early acute stage of illness, i.e., during the pyrexic phase when the virus is replicating. Nasopharyngeal swabs must be placed in a virus transport medium and forwarded immediately to the laboratory at refrigerated temperature. Equine influenza, rhinopneumonitis, and equine viral arteritis are the three viral infections causing outbreaks of respiratory disease in North America. African horse sickness, although foreign to North America, could be introduced despite stringent horse importation regulations. Specific antiviral therapy is not available to treat viral respiratory disease in the horse. A variety of inactivated and modified live vaccines, however, are available to prevent clinical disease and the spread of infection caused by the common viral respiratory pathogens. A considerable amount of research is underway to enhance the potency and duration of immunity of the present vaccines against influenza and rhinopneumonitis. This research is directed at defining and characterizing the importance of specific glycoprotein antigens on the surface of the virus, which trigger the various host immune responses, and determining whether they are stimulatory or suppressive.     

High efficient expression of human endostatin in E.coli and its antiangiogensis activity

AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors.     

五种鬼伞过氧化物酶和酯酶的同工酶研究

应用垂直板聚丙烯酰胺凝胶电泳对五种野生鬼伞 (Coprinus)真菌进行了过氧化物酶 (POD)和酯酶(EST)的同工酶分析 ,结果表明 :五种鬼伞的POD和EST同工酶酶谱比较稳定清晰 ,且分别有一条共同的酶带 ,可能是鬼伞属的POD和EST同工酶特征酶带 ;POD和EST同工酶酶谱均表明 ,家园鬼伞 (C .domesticus)和瓦鳞鬼伞 (C .clavatus)间有较近的亲缘关系 ;不同种鬼伞的POD和EST同工酶之间既有共同的特征 ,又各自有本物种的特有特征 ,POD和EST同工酶酶谱可以作为鬼伞属种类鉴定、亲缘关系比较的重要依据。     

Roles of the Kv1.2, Kv1.5, Kv2.1 potassium channels in hypoxia pulmonary vasoconstriction

AIM: To determine the role of Kv1.2, Kv1.5, Kv2.1 in the hypoxia pulmonary vasoconstriction (HPV). METHODS: Male Wistar rats were divided into two groups: normoxic group and hypoxic group. The single smooth muscle cell was obtained from pulmonary artery of Wistar rats with acute enzymatic digestion method. The conventional whole-cell patch clamp technique was used to record the resting membrane potential (Em) and the potassium currents of voltage-gated potassium channel (IKv) in rat pulmonary arterial smooth muscle cells (PASMC). Intracellular application of Kv1.2/Kv1.5/Kv2.1 antibodies (1∶125) was conducted through the whole-cell patch clamp system. RESULTS: ① Em of PASMC was depolarized after 24 h hypoxia compared with that of control cells . IKv of PASMC was decreased after 24 h hypoxia, . ② The mixture of Kv1.2/Kv1.5/Kv2.1 antibodies depolarized Em and inhibited IKv in PASMC from normoxic rat, whereas the mixture of Kir2.1/Kir2.3/Kir4.1 antibodies had no effects on them. ③ The mixture of Kv1.2/Kv1.5/Kv2.1 antibodies and the mixture of Kir2.1/Kir2.3/Kir4.1 antibodies had no effects on IKv and Em from rats hypoxic for 24 h. CONCLUSION: Kv1.2, Kv1.5, Kv2.1 might be oxygen sensitive potassium channels which mediated HPV.     

In vitro studies of Norwegian Red bovine semen immobilized and cryopreserved in alginate solid gel network

Development of new semen cryopreservation techniques improving sperm survival and ensuring availability of viable spermatozoa for a prolonged time‐period after AI is promising tools to reduce sensitivity of timing of AI and enhance overall fertility. The SpermVital^® technology utilizes immobilization of bull spermatozoa in a solid network of alginate gel prior to freezing, which will provide a gradual release of spermatozoa after AI. The objective of this study was to compare post‐thaw sperm quality and in vitro sperm survival over time of Norwegian Red bull semen processed by the SpermVital^® (SV) technology, the first commercialized production line of SpermVital^® (C) and by conventional procedure applying Biladyl^® extender (B). Post‐thaw sperm motility was not significantly different between SV, C and B semen (p > .05). However, sperm viability and acrosome intactness were higher for SV than C and B semen (p < .05). Small differences in DNA quality were observed (p < .05). Sperm viability after storage in uterus ex vivo was higher for SV than for C semen (p < .05). Furthermore, sperm survival in vitro over time at physiological temperature was significantly higher for SV semen than C semen as well as B semen during the incubation period of 48 hr (p < .05). In conclusion, the SpermVital^® technology is improved and is more efficient in conserving post‐thaw sperm quality and results in higher sperm viability over time in vitro for SV than for C and B semen.     

菱传粉生物学研究

以6 个菱品种为试材，对花器形态特征、群体花期及单花开花动态、开花时间、柱头可授性、花粉活力、杂交指数 及传粉形式等进行观察测定。结果表明：菱花单生，两性，单花花期15 h，群体花期在5～10 月，盛花期在7～8 月；不同 品种在一天中的开花高峰期存在差异；柱头可授期主要在萼片开裂后2 h 内，结实率在50% 以上；花粉在自然授粉条件下活 力可保持7.5 h；以自花授粉为主，同时存在一定程度的无融合生殖和少量异花授粉。     

一种高质量天门冬基因组DNA提取方法

针对天门冬[Asparagus cochinchinensis(Lour.)Merr.]成熟叶片富含多糖、多酚、蛋白质及其他次生代谢物质的特点,采用改良CTAB法对其基因组总DNA进行提取,并对DNA进行质量检测。结果表明,改良CTAB法可从天门冬成熟叶片中获得纯度高、完整性好的总DNA,其OD260 nm/OD280 nm为1.86,OD260 nm/OD230 nm为2.36,浓度为390μg/mL,产率达11.7μg/g,多糖、多酚和RNA杂质被去除干净,无降解现象,说明该方法提取的天门冬基因组总DNA质量较高。     

子午岭黑山羊与辽宁绒山羊产肉性能、肉品质、肌肉营养成分和脂肪酸含量比较

1990年,甘肃省庆阳市开始引入辽宁绒山羊对子午岭黑山羊进行杂交改良,但目前尚不清楚2个品种在脂肪酸含量、肌肉营养成分等方面的差异,影响了杂交改良效果。试验旨在分析两个绒山羊品种的产肉性能、肉品质、肌肉营养成分和脂肪酸含量差异,为绒山羊的杂交改良提供理论依据。本研究选取相同饲养管理条件下、9月龄的子午岭黑山羊和辽宁绒山羊公羊各5只,测定其屠宰性能以及背最长肌、前腿肌和后腿肌处的肉品质、脂肪酸含量和肌肉营养成分。结果表明：子午岭黑山羊的胴体重、屠宰率、净肉重、净肉率、眼肌面积、GR值、剪切力和滴水损失低于辽宁绒山羊（P<0.05）,但其肌肉的平均亮度值、色度值、pH₁和pH₂₄高于辽宁绒山羊（P<0.01）。营养成分测定结果表明,子午岭黑山羊肌肉的水分和粗灰分含量高于辽宁绒山羊（P<0.05）,但肌内脂肪和粗蛋白含量低于辽宁绒山羊。在2个山羊品种的肌肉中均检测到11 种饱和脂肪酸（SFA,以棕榈酸和硬脂酸为主）、10种多不饱和脂肪酸（PUFA,以亚油酸和顺-11,14-二十碳二烯酸为主）和6种单不饱和脂肪酸（MUFA,以油酸为主）,子午岭黑山羊肉中的SFA、PUFA、n-3 PUFA、n-6 PUFA含量和PUFA/SFA值均高于辽宁绒山羊（P<0.01）,但MUFA含量低于辽宁绒山羊（P<0.01）。结果表明,辽宁绒山羊有更高的产肉力,但子午岭黑山羊肌肉品质和营养成分更佳,脂肪酸组成和含量更符合人类健康膳食标准。     

Prevalence of feline herpesvirus 1, Chlamydophila felis, and Mycoplasma spp DNA in conjunctival cells collected from cats with and without conjunctivitis

OBJECTIVE: To use PCR assays to determine the prevalence of feline herpesvirus 1 (FHV-1), Chlamydophila felis, and Mycoplasma spp DNA in conjunctival cells collected from cats with and without conjunctivitis; to compare results of conventional and real-time fluorogenic PCR assays for amplification of FHV-1 DNA; and to determine whether copy numbers of FHV-1 DNA are correlated with conjunctivitis. ANIMALS: 55 cats with active conjunctivitis, 39 healthy cats that never had conjunctivitis, and 32 cats with a history of conjunctivitis that had been resolved for at least 3 months. PROCEDURES: Samples were obtained by rolling cotton-tipped applicators on the ventral conjunctiva of awake cats treated topically with proparacaine. The DNA was extracted from the swab specimens and assessed in PCR assays to detect DNA of FHV-1 (fluorogenic PCR assay and conventional PCR assay), Mycoplasma spp (conventional PCR assay), and C felis (conventional PCR assay). RESULTS: Overall prevalence rates of FHV-1, C felis, and Mycoplasma spp as assessed by the conventional PCR assays were 6.7%, 3.2%, and 9.6%, respectively. Percentage concordance between conventional PCR and fluorogenic PCR assays for FHV-1 was 92.5%. There were no significant differences among the 3 groups of cats for the mean copy number of FHV-1 divided by the copy number of glyceraldehyde-3-phosphate dehydrogenase. CONCLUSIONS AND CLINICAL RELEVANCE: Mycoplasma spp were the most prevalent organism detected and was associated with conjunctivitis. This study could not confirm that there are increased copy numbers of FHV-1 DNA in cats with conjunctivitis, compared with the copy numbers for cats without conjunctivitis.