Rapidly quantitative detection of Nosema ceranae in honeybees using ultra-rapid real-time quantitative PCR

BackgroundThe microsporidian parasite Nosema ceranae is a global problem in honeybee populations and is known to cause winter mortality. A sensitive and rapid tool for stable quantitative detection is necessary to establish further research related to the diagnosis, prevention, and treatment of this pathogen.ObjectivesThe present study aimed to develop a quantitative method that incorporates ultra-rapid real-time quantitative polymerase chain reaction (UR-qPCR) for the rapid enumeration of N. ceranae in infected bees.MethodsA procedure for UR-qPCR detection of N. ceranae was developed, and the advantages of molecular detection were evaluated in comparison with microscopic enumeration.ResultsUR-qPCR was more sensitive than microscopic enumeration for detecting two copies of N. ceranae DNA and 24 spores per bee. Meanwhile, the limit of detection by microscopy was 2.40 × 10⁴ spores/bee, and the stable detection level was ≥ 2.40 × 10⁵ spores/bee. The results of N. ceranae calculations from the infected honeybees and purified spores by UR-qPCR showed that the DNA copy number was approximately 8-fold higher than the spore count. Additionally, honeybees infected with N. ceranae with 2.74 × 10⁴ copies of N. ceranae DNA were incapable of detection by microscopy. The results of quantitative analysis using UR-qPCR were accomplished within 20 min.ConclusionsUR-qPCR is expected to be the most rapid molecular method for Nosema detection and has been developed for diagnosing nosemosis at low levels of infection.     

Immunogenic properties of Landrace pigs selected for resistance to mycoplasma pneumonia of swine

Mycoplasma pneumonia of swine (MPS) lung lesions and immunogenic properties were compared between a Landrace line that was genetically selected for reduced incidence of pulmonary MPS lesions, and a non‐selected Landrace line. The MPS‐selected Landrace line showed significantly lower degrees of pulmonary MPS lesions compared with the non‐selected Landrace line. When changes in immunity before and after vaccination were compared, the percentage of B cells in the peripheral blood of the MPS‐selected Landrace line was significantly lower than that of the non‐selected line. Furthermore, the concentration of growth hormone and the mitogen activity of peripheral blood mononuclear cells in the MPS‐selected Landrace line showed significantly (P < 0.05) lower increases after vaccination than the non‐selected line. Conversely, the concentration of peripheral blood interferon (IFN)‐γ and salivary immunoglobulin A (IgA) after Mycoplasma hyopneumoniae vaccination was significantly higher in the MPS‐selected Landrace line than in the non‐selected line. Gene expression of toll‐like receptor (TLR)2 and TLR4 was significantly higher in the MPS‐selected Landrace line in immune tissues, with the exception of the hilar lymph nodes. The present results suggest that peripheral blood IFN‐γ, salivary IgA TLR2, and TLR4 are important immunological factors influencing the development of MPS lesions.     

Utilization of digital differential display to identify differentially expressed genes related to rumen development

This study aimed to identify the genes associated with the development of the rumen epithelium by screening for candidate genes by digital differential display (DDD) in silico. Using DDD in NCBI's UniGene database, expressed sequence tag (EST)‐based gene expression profiles were analyzed in rumen, reticulum, omasum, abomasum and other tissues in cattle. One hundred and ten candidate genes with high expression in the rumen were derived from a library of all tissues. The expression levels of 11 genes in all candidate genes were analyzed in the rumen, reticulum, omasum and abomasum of nine Japanese Black male calves (5‐week‐old pre‐weaning: n = 3; 15‐week‐old weaned calves: n = 6). Among the 11 genes, only 3‐hydroxy‐3‐methylglutaryl‐CoA synthase 2 (HMGCS2), aldo‐keto reductase family 1, member C1‐like (AKR1C1), and fatty acid binding protein 3 (FABP3) showed significant changes in the levels of gene expression in the rumen between the pre‐ and post‐weaning of calves. These results indicate that DDD analysis in silico can be useful for screening candidate genes related to rumen development, and that the changes in expression levels of three genes in the rumen may have been caused by weaning, aging or both. © 2015 Japanese Society of Animal Science     

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions in Large White pigs selected for higher peripheral blood immune capacity

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions were compared between the immunity‐selected Large White line and the non‐selected Large White line. The selected Large White line showed a higher level of pulmonary MPS lesions compared with the non‐selected Large White line. Subsequent to vaccination, the percentage of natural killer cells and T cells (CD3⁺CD4⁺CD8^‐ and CD3⁺CD4^?CD8⁺ T cells) were significantly increased in the non‐selected line but remained unchanged in the immunity‐selected Large White line. Secretion of Mycoplasma hyopneumoniae vaccine‐specific immunoblogulin G and phagocyte activity in peripheral blood were significantly higher in the immunity‐selected Large White line than in the non‐selected line. Expression of interleukin (IL)‐4 and IL‐6 messenger RNA in hilar lymph nodes was significantly lower in the immunity‐selected Large White line than in the non‐selected line. However, expression of IL‐10 in all immune tissues was significantly higher in the immunity‐selected Large White line. These results suggest that the selection for high immunity was not effective in increasing resistance to MPS lung lesions.     

Apparent ileal digestibility of nutrients and amino acids in soybean meal,fish meal,spray‐dried plasma protein and fermented soybean meal to weaned pigs

This study sought to determine whether fermentation could increase apparent ileal digestibility (AID) of dry matter (DM), nitrogen (N), energy (E) and amino acids (AA) in fermented soybean meal (FSBM) greater than that of soybean meal (SBM) in weaned pigs. Four weaned pigs (10.00 ± 0.30 kg) were surgically equipped with T‐cannulas and randomly followed a 4 × 4 Latin square design of treatments (SBM, FSBM, fish meal and spray‐dried plasma protein). Overall, the fermentation process was able to reduce the amount of anti‐nutritional factors (ANF), including trypsin inhibitors, raffinose and stachyose, in the FSBM diet, which were significantly reduced by 39.4, 92.2, and 92.9%, respectively, as compared to the SBM diet. As a consequence of ANF reduction in FSBM, the AID of DM, N and E as well as AA was significantly greater with FSBM than SBM. Taken all together, the fermentation process improved the nutritional quality of SBM, due to ANF reduction, leading to improvement of digestibility of AA. As such, FSBM can be potentially used as a specialized feed ingredient, especially for young animal diets in an attempt to reduce diet costs.     

Mycoplasma pneumonia of swine (MPS) resistance and immune characteristics of pig lines generated by crossing an MPS pulmonary lesion selected Landrace line and a highly immune capacity selected Large White line

To understand the influence of crossbreeding on Mycoplasma pneumonia of swine (MPS) resistance and immune characteristics, two crossbred lines were characterized. One crossbred line, LaWa, was generated by crossing the MPS pulmonary lesion selected Landrace line (La) and the highly immune‐selected Large White line (Wa). The second crossbred line, LaWb, was generated by crossing the La line and the nonselected Large White line (Wb). The crossbred LbWb line (nonselected Landrace line × nonselected Large White line) and the La line were used as controls. The LaWa and LaWb lines had an intermediate level of MPS lung lesions between La and LbWb lines, although the difference was not statistically significant. After stimulation with sheep red blood cells (SRBCs), the LaWb and LaWa lines showed immune characteristics similar to that of the La line; the number of monocytes in peripheral blood increased, while B cells, T cells, secretion of SRBC‐specific immunoglobulin G, and interleukin (IL)‐13 decreased. Additionally, the number of natural killer (NK) cells and the expression of IL‐4 and IL‐17 were significantly higher in the LaWb and LaWa lines, respectively. These data suggested that crossbreeding of La and Wa lines resulted in the inheritance of some of the selected immune responses.     

高寒地区蛹虫草大米培养基的优化

以引进西藏的6株蛹虫草为试材,对比研究了其菌丝生长速率、出草产出及长势,筛选优良的蛹虫草菌株,并通过正交实验筛选对蛹虫草大米培养基营养成分的最优水平组合。结果表明:蛹虫草"TAAAS1"的菌丝生长速率虽不是最快的,但出草整齐均匀,出草产出高于其它菌株,表明"TAAAS1"菌株是适宜在高寒地区栽培生产的好品种;蛹虫草大米培养基的最优水平组合为蛹粉4g/L,葡萄糖20g/L,硫酸镁2g/L,磷酸二氢钾1.5g/L。     

BY-150型种子包衣机检测控制系统设计

包衣机在批量加工流程中需要针对供种量和进液量等重要参数进行检测,否则无法保证良好的包衣效果。为此,设计了一种包衣机检测控制系统,可实现对供种量和进液量参数信息的检测和控制。同时,需在供种量检测模块的硬件选型和电路设计中采用尽可能多的抗干扰措施保证种子质量的检测精度,在进液量检测模块的设计中通过采用主板信号控制动作启动、传感器检测电路直接控制动作结束的方式保证进液控制的稳定性,并通过液晶触摸屏为用户提供一个简单直观的人机交互方式。整个系统稳定可靠、抗干扰性好,满足预期的精度要求,包衣效果良好。     

联合收割机脱粒装置三维参数化系统设计

以4ZL－3．0型联合收割机脱粒装置为基础，并以Autodesk公司三维造型软件MDT3．0为设计平台，利用AutoLisp及ObjectARX等开发工具，建立了以数据库为中心的脱粒装置三维参数化设计系统。整个系统按照自上而下的并行方式进行设计，并按照装配等级模型对滚筒的零部件进行了装配。此系统保证了整体的协调性和一致性，并可实现脱粒装置及其变形产品零件图及装置图的快速生成。     

Cellular Localization of Rice SUMO/SUMO Conjugates and in vitro Sumoylation Using Rice Components

正Many proteins are regulated by post-translational modifications,such as the reversible covalent attachment of ubiquitin and ubiquitin-like proteins in eukaryotes(Kerscher et al, 2006). Post-translational modification of proteins by the SUMO protein family is involved in diverse cellular processes, including development, hormonal responses, and biotic and abiotic stress signaling(Park et al, 2011). SUMO modification can modulate protein-protein interactions, intracellular localization or the activities of the protein(Gareau and Lima, 2010).