Detection and characterization of leptospiral antigens using a biotin/avidin double-antibody sandwich enzyme-linked immunosorbent assay and immunoblot.

A biotin/avidin double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of antigens of Leptospira interrogans serovars in experimentally inoculated bovine urine samples was evaluated. Immunoglobulin G (IgG) from rabbits immunized with L. interrogans serovar hardjo type hardjobovis sonicated, whole cell, and formalinized-heated antigen preparations were purified by a protein A-superose column coupled to fast protein liquid chromatography, and evaluated for species specificity in the ELISA. The ELISA using each specific IgG detected as few as 10(4) leptospires of the homologous serovar hardjo diluted in phosphate-buffered saline solution with Tween 20 (PBSS-Tween 20). On immunoblot analysis of proteinase-K-digested whole cell leptospiral preparations, each IgG revealed the presence of bands specific to serovar hardjo, suggesting the presence of serovar-specific epitopes on the lipopolysaccharide molecules. The minimum number of cells of heterologous serovars pomona, grippotyphosa, bratislava, icterohaemorrhagiae and copenhageni detected by each ELISA was greater, ranging from 10(6) to 10(7). The common antigenic determinants observed on immunoblot analysis were different for each specific IgG, except for a major cross-reacting, possibly flagellar, protein doublet at approximately 36-36.5 kDa. Leptospires were equally well detected by the ELISA in both bovine urine and PBSS-Tween 20.     

Characterization of and radioimmunoassay for canine thyroglobulin

Canine thyroglobulin (cTg) has been isolated and purified. It has similar electrophoretic patterns as Tg from other mammalian species. The main fraction had a MW of 660,000, whereas also fractions of a MW of approximately 1,300,000 (dimer) and 330,000 (subunit) were present. The iodine content was 0.8 to 1.0 % (w/w). cTg did not cross-react with antibodies against human Tg to a degree that would allow the use of a radioimmunoassay for human Tg for the determination of cTg in serum or plasma. Therefore a polyclonal antiserum was raised against cTg and a homologous radioimmunoassay was developed, which was sensitive (0.4 μg/l) and specific (cross-reactivity in cTg assay of human Tg, goat Tg, T₄, T₃, and DIT < 0.01 %).

Plasma Tg levels in normal dogs of both sexes and aged 3–15 years amounted to 192 ± 73 μg/l (mean ± SD, n=30). There was no relation between plasma Tg and T₄ levels.     

Construction and Development of the Dagangshan Forest Ecosystem Research Station

Established by the former Ministry of Forestry in 1986, Dagangshan Forest Ecosystem Research Station is one of the 14 national key sites in the field of ecosystem research. In this paper, the basic situation of Dagangshan Forest Ecosystem Station is described, including geographic location, natural conditions, biological resources, research conditions, instruments, achievement, prospects etc.     

山洪对房屋的破坏及防患

山洪对房屋的破坏随着生态环境的恶化而加剧,其损失又随人民生活水平的提高而增加。防患房屋遭受山洪破坏刻不容缓。本文客观地总结了山洪破坏房屋具有的区域性、多样性、毁灭性特征,通过分析房屋遭受山洪破坏的沉痼陋习,初步揭示了山洪惩罚人为性大失误的沉痛教训,进而提出防患山洪破坏房屋要非工程措施与工程措施相结合,运用社会、经济、法律的手段,加以减灾引导,加强建房管理,采取开撇水沟、挖水平槽、刨鱼鳞坑、砌护坡墙、打挡水桩等直接有效的工程措施,加速隐患补救,实现减灾目的。     

氨基酸铁在大鼠小肠中的吸收及组织中沉积研究

本试验用50日龄Wistar纯系雄性大鼠,采用体内原位结扎肠段灌注技术和放射性同位素示踪技术,研究了氯基酸螯合铁(以赖氨酸螯合铁和甘氨酸螯合铁为代表)的吸收特点。试验中观察了向结扎肠段灌注含不同形态铁的吸收液后,不同时间点(5、15、30、60、90、120分钟)血液中59Fe比放射性的动态变化;120分钟(试验结束)时不同组织器官中59Fe的放射性大小和总铁含量以及主要血液学指标的变化。试验结果表明:①赖氨酸和甘氨酸都能促进大鼠对铁的吸收。②与相应摩尔浓度的氨基酸与氯化亚铁的混合物相比,赖氨酸螯合铁和甘氨酸螯合铁能更有效地被大鼠吸收。综合分析本试验结果认为,氨基酸螯合铁都是一种优秀的铁添加剂。从本试验结果看,放射性同位素示踪技术结合结扎十二指肠段灌注技术不失为一种研究动物对微量元素吸收情况的理想试验手段。     

茄子早熟育种研究Ⅱ选择指数与早熟性选择

研究结果表明,由性状果实最大周长,果实体积,门茄显蕾期,单株产量等性状构成的选择指数较好;进一步进行遗传相关分析结果认为,应用选择指数时应注意果实体积这个性状,单株产量是一个重要的辅助性状,门茄显蕾期是一个限制性因子。应用选择指数对品种评价结果认为,早熟性较好的品种有:天津早茄、西安绿茄、二民、七叶茄、白线茄和汉中荷包。     

Comparison of dot blot hybridization, polymerase chain reaction, and virus isolation for detection of bovine herpesvirus-1 (BHV-1) in artificially infected bovine semen.

Bovine semen samples spiked with bovine herpesvirus 1 (BHV-1) were used to compare dot blot hybridization, polymerase chain reaction (PCR), and virus isolation for detection of BHV-1 in bovine semen. The PCR amplification used primers targeting the BHV-1 thymidine kinase gene and a nucleic acid releasing cocktail (GeneReleaser); the PCR product was used as the DNA probe in dot blot hybridization; virus isolation was done in primary bovine fetal testis (BFT) cell cultures. Semen diluted 1:20 in tissue culture medium had the least cytotoxicity and inhibition of viral cytopathic effects in BFT cells, allowing detection of 1 TCID50/100 microL of BHV-1 suspension by virus isolation. The presence of foreign DNA such as bovine sperm DNA or salmon sperm DNA increased the sensitivity of dot blot hybridization in detecting BHV-1, allowing detection of 20,000 TCID50/100 microL of neat semen. The inhibition of PCR amplification of BHV-1 DNA in bovine semen was eliminated by diluting the samples 1:20 in tissue culture medium. The best PCR amplification was obtained when semen was diluted 1:20 and when a reaction buffer of pH 9.0, with 1.0 mM MgCl2 was used. Under these conditions, the PCR followed by ethidium bromide staining of agarose gels could detect 1 TCID20/100 microL of sample, whereas PCR followed by Southern blot hybridization could detect 0.01 TCID50/100 microL of sample.(ABSTRACT TRUNCATED AT 250 WORDS)     

Destruction of Trichinella spiralis spiralis during the preparation of the "dry cured" pork products proscuitto, proscuittini and Genoa salami

Genoa salami, proscuittini and proscuitto were prepared from pork carcasses that were heavily infected experimentally with Trichinella spiralis spiralis. Genoa salami was prepared with salt concentrations of 2.0%, 2.75% and 3.3%. Proscuitto was prepared by two procedures approved by Agriculture Canada. At various times postpreparation, samples of the various cured products were taken and examined by pepsin digestion and rat bioassay for the presence of viable trichinae. Water activity and pH of the cured meat were also determined. Curing of the various products was shown to destroy the Trichinella larvae. Pepsin digestion revealed that larvae progressively became loosely coiled, uncoiled and more subject to digestion (ghost larvae) during the curing process. Rat bioassay revealed the presence of viable trichinae in the proscuitto prepared using a sodium chloride salt mixture at day 34 but not at day 48 postpreparation. All other bioassays carried out on Genoa salami between 13 and 42 days postpreparation, on proscuittini between days 27 and 69 and on proscuitto between days 34 and 69 were negative for viable trichinae. Under the conditions of this study, preparing Genoa salami with salt concentrations as low as 2% did not appear to affect the destruction of Trichinella larvae.