甘蔗抗坏血酸过氧化物酶基因(ScAPX)的克隆及表达分析

抗坏血酸过氧化物酶(ascorbate peroxidase,APX)是清除活性氧(reactive oxygen species,ROS)的重要酶类.本研究对针刺接种黑粉菌(Sporisorium scitamineum)后的抗、感基因型甘蔗(Saccharum officinarum)进行APX活性测定.结果表明,甘蔗接种黑穗病菌48h内,抗病品种(崖城05-179) APX活性显著高于感病品种(柳城03-182) (P＜0.05).借助电子克隆技术,结合RT-PCR方法,从甘蔗中分离到一个APX基因,并命名为ScAPX(GenBank登录号:KJ7565501).生物信息学分析显示,ScAPX基因全长l 171bp,包含一个1 038 bp的完整开放阅读框,编码345个氨基酸.ScAPX编码的蛋白不含信号肽,推测其为非分泌蛋白,定位于线粒体基质和叶绿体基质中的概率分别为91.1％和88.7％.实时荧光定量PCR检测结果显示,ScAPX在甘蔗根、芽、叶、蔗皮和蔗髓中均有表达,为组成型表达基因,表达量在蔗皮中最高,叶中最低,前者是后者的19.7倍;在外源因子处理后0～48 h,ScAPX基因的表达受水杨酸(salicylic acid,SA)、茉莉酸甲酯(methyl jasmonate,MeJA)、过氧化氢(hydrogen peroxide,H2O2)、脱落酸(abscisic acid,ABA)、氯化钠(sodium chloride,NaCl)和聚乙二醇(polyethylene glycol,PEG)诱导,SA、MeJA和H2O2处理后的ScAPX 转录本积累量峰值较外源激素(ABA)和环境(NaCl和PEG)胁迫早.前者的ScAPX转录本变化呈现出胁迫初期积累量增加,达到峰值后又逐步下降的特点;在同样的测试时间(处理后24 h)内,ABA、NaCl和PEG处理后ScAPX转录本在达到峰值后未见下降.虽然在外源胁迫下,ScAPX表达量变化存在明显差异,但其表达均表现出正响应上述外源因子对甘蔗的胁迫.本研究为后续深入鉴定该基因的功能与进一步应用提供基础资料.     

植物MYB转录因子的同义密码子使用偏好性分析

前人研究显示MYB（V-myb avian myeloblastosis viral oncogene homolog）转录因子对应答植物逆境胁迫等有重要作用,甘蔗MYB转录因子的研究报道并不太多。通过对与甘蔗亲缘关系近的禾本科作物玉米、高粱和水稻MYB转录因子基因家族的密码子使用偏好性进行比较分析。结果表明,3个物种的MYB转录因子的蛋白质序列上存在高度的保守性,仅有部分密码子使用不太一样,此外,分析还显示基因碱基组成有可能在一定程度上影响了密码子的偏好使用。     

Foot and Mouth Disease and Livestock Husbandry Practices in the Adamawa Province of Cameroon

Foot and mouth disease (FMD) is a highly contagious viral disease of even-toed ungulates and is endemic in most of the tropics. A cross-sectional study using a stratified, two-stage random sample design was undertaken in the Adamawa Province of Cameroon. The objectives were to measure the reported herd-level prevalence of FMD and a range of husbandry practices important for its transmission. The owner-reported prevalence for the previous 12 months was 57.9% (50.4–65.4%), although there was a significant variation across the Province. During the previous dry season, 46.5% (38.6–54.4%) of herds had gone on transhumance. Herds had high numbers of contacts with other herds while on transhumance (98.6%), at pasture (95.8%) and at night (74.4%), with medians of 7–10, 4–6 and 1–3 daily contacts, respectively. The high level of endemic FMD and potential for disease spread presents a significant challenge for control and eradication. Locally sustainable methods need to be developed upon which larger regional control programmes could be built in the future.     

Determination of the clonal structure of avian Escherichia coli strains by isoenzyme and ribotyping analysis

Forty-nine avian Escherichia coli strains isolated from different outbreak cases of septicemia (24), swollen head syndrome (14) and omphalitis (11), and 20 strains isolated from poultry with no signs of the mentioned illnesses, for a total of 69 strains, were typed by isoenzyme profile and ribotyping analysis by restriction fragment length polymorphism (RFLP). Isoenzyme analysis discriminated better among strains (0-0.07 degree of genetic dissimilarity) than ribotyping analysis (0- 0.02 degree of genetic dissimilarity). The enzyme profiles of the E. coli isolates allowed the identification of 33 clones that were organized into six main clusters (A-F). Cluster A comprised 87% of the pathogenic strains and had no commensal strains, while commensal strains were assigned to clusters B-F. The ribotyping analysis resulted in a more heterogenous distribution of strains but most of those that cause the same type of infection were kept close together. Taken as a whole, these results demonstrate that pathogenic clones are more similar to one another when compared with commensal strains and suggest a correlation between the genetic background and the pathogenic characteristics of avian pathogenic E. coli strains.     

Evidence of an association between use of anti-microbial agents in food animals and anti-microbial resistance among bacteria isolated from humans and the human health consequences of such resistance

Several lines of evidence indicate that the use of anti-microbial agents in food animals is associated with anti-microbial resistance among bacteria isolated from humans. The use of anti-microbial agents in food animals is most clearly associated with anti-microbial resistance among Salmonella and Campylobacter isolated from humans, but also appears likely among enterococci, Escherichia coli and other bacteria. Evidence is also accumulating that the anti-microbial resistance among bacteria isolated from humans could be the result of using anti-microbial agents in food animals and is leading to human health consequences. These human health consequences include: (i) infections that would not have otherwise occurred and (ii) increased frequency of treatment failures and increased severity of infection. Increased severity of infection includes longer duration of illness, increased frequency of bloodstream infections, increased hospitalization and increased mortality. Continued work and research efforts will provide more evidence to explain the connection between the use of anti-microbial agents in food animals and anti-microbial-resistant infections in humans. One particular focus, which would solidify this connection, is to understand the factors that dictate spread of resistance determinants, especially resistant genes. With continued efforts on the part of the medical, veterinary and public health community, such research may contribute to more precise guidelines on the use of anti-microbials in food animals.     

甘蔗抗感黑穗病池的构建和抗病基因分子标记

 由甘蔗黑穗病菌(Ustilago scitaminea Syd)引起的甘蔗黑穗病是一种世界性的主要病害。在世界各植蔗区普遍发生,造成感病品种蔗茎产龟和蔗糖分的严重损失,在旱地甘蔗上危害更为严重。公认控制该病经济有效的途径是种植抗病品种。     

Between prepartum luteolysis and onset of expulsion

In the cow the foetal endocrine signals that initiate the calving process result in prepartum luteolysis. Withdrawal of progesterone (P4) action is a prerequisite for a normal calving. The rather abrupt declining influence of P4 is followed by a cascade of physiological processes in the myometrium and cervix. This contribution will focus on some of these events. Like in many other species, the myometrium in cows is not completely inactivated during pregnancy. So-called contractures have been registered during the final weeks of gestation and their EMG-characteristics in cows show a low frequency (on average: 13.6 per day) and long duration (on average 12.1 min). They are not evenly spread over the day because they occur less frequently when the cows are disturbed for feeding or cleaning their stables. Contractures affect several foetal functions. In the cow these contractures disappear during a period of about 8-9h when maternal plasma P4 levels are rapidly declining before calving. There is experimental evidence that this temporary inhibition is associated with prepartal luteal regression. The cause of this inhibition is still unknown. Because nitrous oxide inhibits smooth muscle cells and evidence in laboratory animals indicates that expression of the inducible form of nitrous oxide (iNOS) is downregulated in myometrium, but upregulated in the cervix around the onset of parturition, we started to investigate the role of this enzyme in bovine tissues around calving. By means of a RT-PCR technique, we obtained a first indication that iNOS is hardly expressed in the myometrium during calving, while expression was clearly detected at day 4 after calving. Analysis of prepartum en periparturient biopsies from myometrium and cervix with quantitative PCR is still underway. In six pregnant cows, provided with uterine EMG-electrodes and with ultrasonic crystals implanted on the caudal cervical rim to measure cervical dilatation, calving was induced with an injection of prostaglandin (PG) F2alpha. While maternal plasma P4 levels had significantly declined within 8h after PG treatment, the myometrium escaped from temporary inhibition with the development of a parturient contractility pattern on average at 13.5h after injection. However, it was only at 28 h after PG treatment that the first sustained increase of the opening of the vaginal ostium of the cervix was measured.     

Induction of protective immunity in chickens immunised with plasmid DNA encoding infectious bursal disease virus antigens

Direct DNA inoculations were used to determine the efficacy of gene immunisation of chickens to elicit protective immune responses against infectious bursal disease virus (IBDV). The vp2 gene of IBDV strains GP40 and D78, and the vp2-vp4-vp3 encoding segment of strain D78 were cloned in an expression vector which consisted of human cytomegalovirus (HCMV) immediate early enhancer and promoter, adenovirus tripartite leader sequences and SV40 polyadenylation signal. For purification of vaccine-quality plasmid DNA from E. coli, an effective method was developed. Chickens were vaccinated by inoculation of DNA by two routes (intramuscular and intraperitoneal). Two weeks later, chickens were boosted with DNA, and at 2 weeks post-boost, they were challenged with virulent IBDV strain. Low to undetectable levels of IBDV-specific antibodies and no protection were observed with DNA encoding VP2. However, plasmids encoding VP2-VP4-VP3 induced IBDV-specific antibodies and protection in the chickens. DNA immunisation opens a new approach to the development of gene vaccines for chickens against infectious diseases.