Use of a genetically attenuated strain of Aeromonas salmonicida to vaccinate salmonid fish

A genetically attenuated strain of Aeromonas salmonicida has been developed that has a complete deletion of the aroA gene (Brivax II), making it suitable for development as a commercial vaccine. Brivax II was effectively cleared from Atlantic salmon, Salmo salar, a species highly susceptible to furunculosis, confirming that it is attenuated. Clearance rate was dependent on the vaccine dose administered, being longer with higher doses. Immunological studies using Brivax II injected in rainbow trout, Oncorhynchus mykiss, confirmed that live vaccines stimulate a greater response, in terms of generating leucocytes able to proliferate to a subsequent encounter with antigen, relative to killed vaccines. Development of strains of Brivax II as carriers of heterologous antigens was also investigated. Escherichia coli -galactosidase was chosen as the model antigen, and three strains containing plasmids with the LacZ gene were constructed (Brivax 12, Brivax 61 and Brivax 107). All three strains were shown to express -galactosidase in vivo in rainbow trout and to be cleared effectively. Interestingly, Brivax 107 was cleared faster than the other two Lac⁺ strains and had the highest level of -galactosidase activity. The two strains expressing lower levels of activity also behaved differently in vivo, in that Brivax 12 accumulated derivatives expressing lower levels of -galactosidase activity, suggesting that mutants are being selected in vivo. The potential advantages of live vaccines over killed vaccines are discussed.     

Age‐related histomorphometric and immunohistochemical changes of the moderator band in Egyptian Baladi cattle

The moderator band (MB) is a common fibromuscular anatomical structure for the right ventricle of most animals. The histomorphometric and immunohistochemical analysis of the MBs of Egyptian Baladi cattle in relation to age was the aim of this study. Eighteen clinically healthy animals of both sexes were used for this study. The animals were divided into three groups depending on age, group I (N = 4, <1 year), group II (N = 8, 1–2 years) and group III (N = 6, 4–8 years). Cross sections of the MBs from all groups were stained with H&E, Masson's trichrome and anti‐connexin43 (Cx43) antibody for histological and immunohistochemical examinations. Also, measurements for the thickness of the endocardium of the MB as well as, the wall of its muscular artery were conducted. Bundles of Purkinje fibres (PFs) were identified peripherally in the endocardial layer and among the myocardial fibres in the core of each MB. The infiltration of endocardial adipocytes was the characteristic for MBs of old animals. All morphometric data showed a significant increase with the advancement of age. Immunohistochemical findings revealed the localization and distribution of Cx43 in the PFs and intercalated discs of all examined MBs. However, variation of Cx43 immunoreactivity was found among the groups depending on the age. On the basis of this study, this conclusion of different histomorphometry and Cx43 expression of the MBs in relation to age was drawn. These interesting findings provide further insight into age‐related physiological and pathological heart conditions.     

阿根廷利索公司大豆根瘤菌剂对大豆结瘤及产量的影响

[目的]验证阿根廷利索公司大豆根瘤菌剂与北方大豆主栽品种的结瘤能力。[方法]以黑河43号为供试品种,采用完全随机区域排列方式,设5个处理,每个处理用不同的大豆根瘤菌剂(Rizoliq Top、Rizoliq Top+Premax、Signum+Premax-Signum、LLI+Premax-LLI)进行拌种,调查大豆植株的生长性状和小区产量。[结果]Rizoliq Top菌剂数量为国家标准最低标准的5.2倍;包衣后种子带菌量为5.48×104CFU/m L;菌剂Rizoliq Top+Premax和LLI+Premax-LLI在荚数、粒数、百粒重方面与对照相比差异不显著,但均高于对照,产量显著高于对照,分别增产14.32%和22.96%。[结论]大豆根瘤菌Rizoliq Top+Premax和LLI+Premax-LLI与黑河43号相互适配性好,增产效果显著。Signum+Premax-Signum和Rizoliq Top与黑河主栽品种黑河43号大豆适配性差或不适应黑河环境,导致增产效果不理想。     

TiC/ZA43复合材料耐磨性能的试验研究

采用原位接触反应法制备了ＴｉＣ／ＺＡ４３复合材料，并对其在干摩擦和预先滴油润滑条件下的摩擦摩损性能进行了试验研究，同时还用扫描电子显微镜对试样磨损表面形貌进行了观察，进而对材料的磨损机理作为分析与讨论，结果表明，随着ＴｉＣ质量分数的增大，ＴｉＣ／ＺＡ４３复合材料的耐磨性能提高，ＺＡ４３合金的磨损机理是以严重的犁削和磨损为主，而ＴｉＣ／ＺＡ４３复各材料的磨损机理则以轻微的犁削和氧化磨损为主。     

生长相关蛋白免疫反应神经纤维在20胚龄鸡端脑中的分布

为了给家禽发育神经解剖学积累资料,应用免疫组化 Ａ Ｂ Ｃ 法,对 ２０ 胚龄鸡端脑进行了生长相关蛋白（ Ｇ Ａ Ｐ４３）免疫反应神经纤维分布的研究。结果表明,在端脑新纹状体中含有大量较细的阳性纤维,阳性纤维多集中在新纹状体靠近背髓板区域,纤维有分支且交织成网。部分阳性纤维穿过背髓板向腹下方移行,伸向旧纹状体扩大部。在旧纹状体扩大部,有呈背腹方向排列的短阳性纤维束,数量较多,向尾侧伸向初级旧纹状体。在初级旧纹状体中,阳性纤维束以腹内侧方向排列为主,并有少量细的纤维分支互相交织,向腹内侧移行,逐渐聚集成纹体丘脑束。纹体丘脑束参与形成额丘脑束,伸向丘脑。     

维生素E通过间隙连接蛋白Cx43对牛颗粒细胞凋亡及增殖的影响

本研究旨在研究维生素E对间隙连接蛋白Cx43的影响和其对牛颗粒细胞增殖与凋亡的作用及其机制。将从牛卵巢中分离获得颗粒细胞进行体外培养,用不同浓度的维生素E (0、25、50、100、200、500 μmol/L)处理细胞24 h。采用流式细胞术检测颗粒细胞的凋亡率、实时荧光定量PCR法检测凋亡标志基因BCL2/BAX、P53及Cx43 mRNA的表达变化、CCK8法检测颗粒细胞的增殖变化。结果表明:流式细胞术检测显示,与对照组相比,体外培养过程中添加100 μmol/L维生素E可显著抑制颗粒细胞凋亡(P<0.05),实时荧光定量PCR检测显示维生素E能引起BCL2/BAX、P53及Cx43基因表达显著变化(P<0.05),CCK8检测显示维生素E处理细胞24和36 h时可以显著促进颗粒细胞增殖(P<0.05)。此研究结果为深入解析维生素E通过影响颗粒细胞增殖、凋亡进而影响卵母细胞发育、成熟的机制及提高雌性动物繁殖能力提供了理论依据。     

猪伪狂犬病病毒四川株的分离鉴定及UL43基因序列分析

从四川某猪场发病仔猪体内分离到1株病毒,该毒株能在Vero、MDBK、PK15、ST、MDCK、BHK21、Marc145、鸡胚成纤维细胞(CEF)上增殖并产生细胞病变。通过蚀斑克隆对其进行纯化,克隆毒株在Vero细胞上为3.0×107TC ID50/0.1mL。该病毒在Vero细胞上连传15代,其TCID50变化很小。病毒对5-溴脱氧尿核苷、氯仿敏感。用该病毒接种家兔和仔猪均出现典型的伪狂犬病症状,gE-ELISA检测接种分离毒的仔猪血清,伪狂犬病毒(PRV)抗体阳性。电镜观察可见直径110～140 nm的典型疱疹病毒粒子。上述结果表明分离株为PRV,并命名为SE株。根据已发表的UL43序列设计1对引物,以分离毒株基因组DNA为模板进行PCR扩增,对目的产物进行克隆及测序分析,结果与GenBank收录的其他PRV毒株(Becker、Ea)的UL43核苷酸序列同源性为98.8%,氨基酸同源性分别为95.2%、97.3%。     

牛坏死杆菌的43 ku外膜蛋白在其黏附细胞中的作用初步分析

旨在明确牛坏死杆菌(Fusobacterium necrophorum)的43 ku外膜蛋白(43K OMP)在其黏附细胞中的作用,本研究将重组43K OMP蛋白和天然43K OMP蛋白与鼠乳腺上皮细胞共孵育,通过免疫荧光方法观察牛坏死杆菌43K OMP对鼠乳腺上皮细胞的黏附作用;同时利用天然蛋白竞争试验和抗体抑制试验明确43K OMP是否介导牛坏死杆菌与细胞的黏附,进一步利用牛坏死杆菌43K OMP基因缺失菌,评价43K OMP基因缺失对牛坏死杆菌黏附力的影响,阐明43K OMP在牛坏死杆菌黏附细胞中的作用。免疫荧光试验结果显示：天然43K OMP和重组43K OMP均能黏附于鼠乳腺上皮细胞表面,天然43K OMP与鼠乳腺上皮细胞或鼠肝细胞预孵育后,牛坏死杆菌黏附数量明显下降(P＜0.05);牛坏死杆菌与43K OMP多抗或单抗预孵育后,黏附于鼠乳腺上皮细胞或鼠肝细胞的牛坏死杆菌数量显著降低(P＜0.05);与牛坏死杆菌A25菌株相比,基因缺失菌A25Δ43K OMP黏附宿主细胞能力极显著下降(P＜0.01),黏附率分别降低了94.4%和90.4%。因此,43K OMP在牛坏死杆菌黏附细胞中发挥关键性作用,深入研究其黏附机理将为揭示牛坏死杆菌致病机制提供理论基础。     

Prenatal and neonatal exposure to the antiandrogen flutamide alters connexin 43 gene expression in adult porcine ovary

Connexin 43 (Cx43) is the predominant gap junction protein within porcine ovary and is required for proper follicle and corpus luteum (CL) development. Recent research suggests maternally or neonatally mediated effects of antiandrogens on reproductive function during adulthood, notably those dependent on gap junctional communication. The current study was conducted to determine whether late gestational or neonatal exposure to the antiandrogen flutamide influences Cx43 gene expression in the adult porcine ovary. Flutamide was injected into pregnant gilts between days 80 and 88 of gestation and into female piglets between days 2 and 10 posnatally. After animals reached sexual maturity, the ovaries were collected from treated and nontreated (control) pigs. Expression of Cx43 mRNA and protein was determined for preantral and antral follicles and for CLs. In addition, 3β-hydroxysteroid dehydrogenase (3β-HSD) expression and progesterone concentration were determined for luteal tissues. In preantral follicles, Cx43 mRNA was down-regulated (P < 0.01) following maternal and neonatal flutamide exposure. In large antral follicles, Cx43 mRNA was up-regulated (P < 0.01) after neonatal flutamide administration. Immunofluorescence showed that Cx43 expression decreased (P < 0.001) in preantral follicles and increased (P < 0.001) in large antral follicles following flutamide exposure. In luteal tissues, Cx43 and 3β-HSD expression and progesterone concentration decreased (P < 0.01) after postnatal flutamide treatment. Overall, these results suggest the involvement of androgens in the regulation of Cx43 expression in pig ovary. Moreover, alteration of Cx43 expression by the administration of flutamide during particular prenatal and neonatal time periods may affect porcine follicle development, as well as CL formation and function.     

Production of specific antibodies against SARS-coronavirus nucleocapsid protein without cross reactivity with human coronaviruses 229E and OC43

Severe acute respiratory syndrome (SARS) is a life-threatening disease for which accurate diagnosis is essential. Although many tools have been developed for the diagnosis of SARS, false-positive reactions in negative sera may occur because of cross-reactivity with other coronaviruses. We have raised polyclonal and monoclonal antibodies (Abs) using a recombinant form of the SARS virus nucleocapsid protein. Cross-reactivity of these anti-SARS Abs against human coronavirus (HCoV) 229E and HCoV OC43 were determined by Western blotting. The Abs produced reacted with recombinant SARS virus nucleocapsid protein, but not with HCoV 229E or HCoV OC43.