Cloning of the acetylcholinesterase 1 gene and identification of point mutations putatively associated with carbofuran resistance in Nilaparvata lugens

Molecular mechanisms of carbofuran resistance in the brown planthopper, Nilaparvata lugens Stål, were investigated. A carbofuran-resistant strain (CAS) showed approximately 45.5- and 15.1-fold resistance compared with a susceptible strain (SUS) and a non-selected field strain (FM), respectively. Activities of the esterase and mixed-function oxidase were approximately 2.8- and 1.6-fold higher, respectively, in the CAS strain than in the SUS strain, suggesting that these enzymes play a minor role in carbofuran resistance. Interestingly, the insensitivity of acetylcholinesterase (AChE) to carbofuran was approximately 5.5- and 3.7-fold higher in the CAS strain compared to the SUS and FM strains, respectively, indicating that AChE insensitivity is associated with carbofuran resistance. Western blot analysis identified two kinds of AChEs, of which the type-1 AChE (encoded from Nlace1, which is paralogous to the Drosophila AChE gene) was determined to be the major catalytic AChE in N. lugens. The open reading frame of Nlace1 is composed of 1989 bp (approximately 74 kD) and revealed 52.5% and 24.3% amino acid sequence identities to those of Nephotettix cincticeps and Drosophila melanogaster, respectively. Screening of point mutations identified four amino acid substitutions (G119A, F/Y330S, F331H and H332L) in the CAS strain that likely contribute to AChE insensitivity. The frequencies of these mutations were well correlated with resistance levels, confirming that they are associated with reduced sensitivity to carbofuran in N. lugens. These point mutations can be useful as genetic markers for monitoring resistance levels in field populations of N. lugens.     

Alteration of helper T-cell related cytokine production in splenocytes during Trichinella spiralis infection

Infection by Trichinella spiralis takes place in two distinct phases: one is the intestinal phase and the other is the muscle phase. To evaluate alterations in cytokine production during a T. spiralis infection, we periodically assessed the cytokine production of splenocytes in mice after infection (AI). The levels of Th2-related cytokines immediately increased after the initiation of T. spiralis larval intestinal invasion (1 week AI). These early elevations in the Th2 response might be associated with the innate immune responses of intestine epithelial cells against T. spiralis larval invasion. IL-4 and IL-13 levels reached a peak prior to the initiation of nurse cell formation (2 weeks AI). Additionally, all Th17-related cytokines, except for IL-17, increased slightly until 2 weeks AI. However, expression levels for all of the Th2 and Th17-related cytokines began to decrease after the initiation of nurse cell formation and reached basal levels at 4 weeks AI, except for IL-5. At the same time, the CD4(+)CD25(+)Foxp3(+) T (regulatory T, T(reg)) cell population increased significantly in the spleen. Additionally, the number of cells in the peripheral lymph nodes increased. In conclusion, T. spiralis larva intestinal invasion induced the production of Th2 and Th17 cell-related cytokines, and the cytokines decreased with T(reg) cell-related cytokine.     

种植海拔高度和采制时期对黄山“松谷”毛峰茶功能成分含量的影响

分析不同种植海拔高度及采制时期对黄山松谷毛峰茶中茶多酚、游离氨基酸、咖啡碱、茶多糖和硒含量的影响;采用相关国家标准对黄山松谷毛峰茶中相应功能成分进行含量测定;从种植海拔高度看,茶样中茶多酚、游离氨基酸含量、酚氨比值及硒含量随种植海拔高度的增加而增加,对于咖啡碱,高海拔茶样中的含量显著高于平地地区茶样,而不同海拔高度的茶样中茶多糖含量差异不显著;从采制时期看,茶样中茶多酚含量为5月最高,4月次之,6月最低;游离氨基酸含量4月最高,5月次之,6月最低;茶样酚氨比值6月最高,5月次之,4月最低;咖啡碱含量4月和5月基本相同,均高于6月;氨碱比值4月最高,5月次之,6月最低;茶多糖含量6月最高,5月次之,4月最低;硒含量4月最高,5月和6月基本一致。此研究结果表明种植海拔高度及采制时期对黄山毛峰茶中茶多酚、游离氨基酸、咖啡碱、茶多糖和硒含量有交互性影响。     

Transient postoperative glycosuria after isoflurane exposure in two dogs

The present report describes two surgical cases involving the development of sudden glycosuria after isoflurane anaesthesia, despite the dogs having normal blood glucose levels and renal glucose reabsorption. The glycosuria manifested 1 day after surgery and resolved spontaneously within 2 days in both cases. Considering that the surgeries (subcutaneous mandibular mass removal and fracture repair) were unrelated to the kidneys, and there were no remarkable events during anaesthesia, the glycosuria may have been associated with the isoflurane anaesthesia. There have been several previous reports of glycosuria in human patients following transient proximal tubule dysfunction due to volatile anaesthetics. This case report suggests the possibility of transient renal dysfunction following isoflurane anaesthesia in these two clinically healthy dogs. However, considering the observational nature of this report, it can not be excluded that any other procedure performed in these animals was responsible of the observed glycosuria.     

遵义朝天椒原种、良种繁育技术规程

本文对朝天椒原种、良种繁育技术措施作了详细介绍和规定,以指导遵义朝天椒育种。     

Pathogenic and immunogenic responses in turkeys following in ovo exposure to avian metapneumovirus subtype C

Commercial turkey eggs, free of antibodies to avian metapneumovirus subtype C (aMPV/C), were inoculated with aMPV/C at embryonation day (ED) 24. There was no detectable effect of virus inoculation on the hatchability of eggs. At 4 days post inoculation (DPI) (the day of hatch (ED 28)) and 9 DPI (5 days after hatch), virus replication was detected by quantitative RT-PCR in the turbinate, trachea and lung but not in the thymus or spleen. Mild histological lesions characterized by lymphoid cell infiltration were evident in the turbinate mucosa. Virus exposure inhibited the mitogenic response of splenocytes and thymocytes and upregulated gene expression of IFN-γ and IL-10 in the turbinate tissue. Turkeys hatching from virus-exposed eggs had aMPV/C-specific IgG in the serum and the lachrymal fluid. At 3 week of age, in ovo immunized turkeys were protected against a challenge with pathogenic aMPV/C.     

Enhancing effects of serum-rich and cytokine-supplemented culture conditions on developing blastocysts and deriving porcine parthenogenetic embryonic stem cells

The present study was conducted to develop an effective method for establishment of porcine parthenogenetic embryonic stem cells (ppESCs) from parthenogenetically activated oocyte-derived blastocysts. The addition of 10% fetal bovine serum (FBS) to the medium on the 3rd day of oocyte culturing improved the development of blastocysts, attachment of inner cell masses (ICMs) onto feeder cells, and formation of primitive ppESC colonies. ICM attachment was further enhanced by basic fibroblast growth factor, stem cell factor, and leukemia inhibitory factor. From these attached ICMs, seven ppESC lines were established. ppESC pluripotency was verified by strong enzymatic alkaline phosphatase activity and the expression of pluripotent markers OCT3/4, Nanog, and SSEA4. Moreover, the ppESCs were induced to form an embryoid body and teratoma. Differentiation into three germ layers (ectoderm, mesoderm, and endoderm) was confirmed by the expression of specific markers for the layers and histological analysis. In conclusion, data from the present study suggested that our modified culture conditions using FBS and cytokines are highly useful for improving the generation of pluripotent ppESCs.     

Development of a novel enzyme-linked immunosorbent assay to detect anti-IgG against swine hepatitis E virus

Swine hepatitis E virus (HEV) is widespread throughout pigs in both developing and industrialized countries. This virus is an important zoonotic agent and a public concern worldwide. Infected pigs are asymptomatic, so diagnosing swine HEV relies on detection of the virus or antibodies against the virus. However, several obstacles need to be overcome for effective and practical serological diagnosis. In this study, we developed an enzyme-linked immunosorbent assay (ELISA) that used a purified recombinant capsid protein of swine HEV. The potential clinical use of this assay was evaluated by comparing it with a commercial kit (Genelabs Technologies, Diagnostics, Singapore). Results of the ELISA were highly correlated with those of the commercial kit with a sensitivity of 97% and specificity of 95%. ROC (receiving operator characteristic) analysis of the ELISA data produced a value of 0.987 (95% CI, 0.977~0.998, p < 0.01). The cut-off value for the ELISA was also determined using negative pig sera. In summary, the HEV-specific ELISA developed in the present study appears to be both practical and economical.     

碳氮比对热带地区鸡粪和蔗渣堆肥腐熟进程的影响

为了探究热带地区鸡粪和蔗渣不同配比对堆肥腐熟进程的影响,寻求最佳物料配比,本研究采用好氧堆肥的方法,研究堆肥过程中不同C/N对堆肥温度、NH₃排放、养分含量及腐熟度等理化指标的影响,探究不同C/N下畜禽粪便的堆肥效果。结果表明,C/N为20 (T1)、30 (T2)和40 (T3)处理的高温持续时间(>50℃)以上的持续时间分别为38天、32天和20天;T1处理的NH₃累积排放量显著高于其他2个处理,T2处理的NH₃挥发速率第1周内最快,说明有机质降解速度最快。随着C/N提高,其有机质含量越高,但NPK总养分含量越低。T2和T3的处理在OM、pH、NPK总养分、GI和NH₄⁺含量指标均达到堆肥标准,而T1处理堆肥结束时OM和NH₄⁺含量不符合标准。综合各个指标,C/N为30的处理为该研究中的最佳堆肥比例。     

基于VAR模型的农业贷款、财政支农对农民收入增长的动态性分析——以河北为例

利用河北1978-2008年农业贷款、财政支农投入和农民收入的实际数据,通过平稳性检验、协整检验和格兰杰非因果检验,进行脉冲分析和方差分解,结果显示:财政支农投入和农业货款与农民收入增长之间存在长期稳定的关系,但其增长作用不大.1978-2008年河北财政支农支出和农业贷款对农民收入的提高的没有起到应有的作用.财政支农...