Discovery and directed evolution of a glyphosate tolerance gene

The herbicide glyphosate is effectively detoxified by N-acetylation. We screened a collection of microbial isolates and discovered enzymes exhibiting glyphosate N-acetyltransferase (GAT) activity. Kinetic properties of the discovered enzymes were insufficient to confer glyphosate tolerance to transgenic organisms. Eleven iterations of DNA shuffling improved enzyme efficiency by nearly four orders of magnitude from 0.87 mM-1 min-1 to 8320 mM-1 min-1. From the fifth iteration and beyond, GAT enzymes conferred increasing glyphosate tolerance to Escherichia coli, Arabidopsis, tobacco, and maize. Glyphosate acetylation provides an alternative strategy for supporting glyphosate use on crops.     

Recovery of Salmonella enterica from seropositive finishing pig herds

The aim of this study was to assess the probability of detecting Salmonella from pen faecal samples in seropositive classified finishing pig herds. The study involved 77 herds from Denmark (20), The Netherlands (20), Greece (17) and Germany (20). The serological herd status was determined by the blood-sampling of 50 finishing pigs. Bacteriological sampling was performed by 20 pen faecal samples per herd. Over-all, 47% of the blood samples had an OD% larger than 10 and 23% larger than 40. Salmonella was isolated from 135 (9.3%) pen faecal samples in 32 herds (42%). Twenty-eight of these herds (87.5%) had a within-herd seroprevalence larger than 50% at sample cut-off OD% > 10. In our study, there was an increasing probability of recovering Salmonella with increasing within-herd seroprevalence. However, this was only a moderate correlation. A correlation coefficient of 0.62 was found between the proportion of culture positive- and seropositive samples in a herd at cut-off OD%> 10 and of 0.58 at cut-off OD% > 40. Serology is a measure of historical exposure, which may or may not correlate closely to the microbiological burden at the time of sampling. Due to the low sensitivity of culture methods, apparent 'false-positive' serological results may well represent real infections not detected by bacteriological testing. For screening purposes, serological testing provides an indication of exposure to Salmonella, which forms the basis for targeted sampling, intervention and logistic slaughter procedures.     

香港土壤研究 Ⅲ.土壤中多环芳烃的含量及其来源初探

通过对香港地区53个土壤样品多环芳烃(PAHs)含量的分析,并利用GIS空间数据管理功能,揭示了香港土壤中PAHs的含量和空间分布特征,同时,运用比值法定性地判断了土壤中PAHs的来源。研究结果表明:香港土壤中可以检测出16种美国环保署规定的优控PAHs中的15种(二苯并(a,h)蒽未被检测到),郊野土壤中PAHs的平均含量为34.2±16.0μg kg-1,而城区土壤中PAHs的平均含量为169±123μg kg-1,港岛动植物公园土壤中的苯并(a)芘的含量最高达到了47.2μg kg-1。在PAHs的来源上,前者的PAHs可能主要来自山火焚烧,而后者可能主要与城区的汽车尾气排放有密切的联系。     

Experimental studies of the role of the little raven (Corvus mellori) in surveillance for West Nile virus in Australia

Objective To study the potential role of an Australian corvid, the little raven (Corvus mellori), in the surveillance for exotic West Nile virus (WNV) in Australia. Method In a series of trials, little ravens were infected with WNV (strain 4132 New York 1999) and Kunjin virus (strain K42886) by the intramuscular route. They were observed for 20 days during which blood and swab samples were taken for virus isolation. Tissue samples were taken from ravens humanely killed during the acute infection period, and at the termination of the trials, for virus isolation, histopathology and immunohistochemistry. Results Ravens infected with WNV became mildly ill, but all recovered and seroconverted. Blood virus titres peaked around 3 to 4 days after inoculation at levels between 10^3.0 to 10^7.5 plaque forming units/mL. Virus or viral antigen was detected in spleen, liver, lung, kidney, intestine, testis and ovary by virus isolation and/or immunohistochemistry. WNV was detected in oral and cloacal swabs from 2 to 7 days post inoculation. The molecular and pathogenic characteristics of the inocula were consistent with them being of high virulence, as expected for this isolate. Ravens infected with Kunjin virus developed viraemia and seroconverted, although they did not develop disease. Conclusions Little ravens do not develop severe disease in response to virulent WNV infection and for this reason may not be important sentinel hosts in the event of an outbreak of WNV, as in North America. However, as they have relatively high viraemias, they may be able to support virus cycles.     

Concomitant turkey herpesvirus-infectious bursal disease vector vaccine and oil-adjuvanted inactivated Newcastle disease vaccine administration: consequences for vaccine intake and protection

Hatchery vaccination protocols in day-old chicks are designed to provide early priming and protection against several poultry diseases including, but not limited to, Marek's disease (MD), infectious bursal disease (IBD), and Newcastle disease (ND). The constraint of concomitant administration of live MD and IBD vaccines plus ND inactivated oil-adjuvanted vaccines (IOAVs) requires improvements in vaccine technology. Single-needle concomitant subcutaneous (SC) application of IBD/MDV and killed NDV vaccine and the use of viral vectors for expression of immunogenic proteins are a current trend in the industry. The objective of this work was to assess the compatibility of a turkey herpesvirus (HVT)-infectious bursal disease (vHVT-IBD) vector vaccine applied simultaneously with IOAV and to evaluate the consequences for vaccine intake, the need for additional immunizations with the respective vaccines, and protection. Five separate trials were performed using double- and/or single-needle injectors. The levels and persistence of vaccine intake, serologic response, vHVT-IBD virus combination with the MD Rispens strain, and/or live NDV vaccination were also assessed. Histopathology and PCR at injection sites showed adequate vaccine intake detected up to 44 days postvaccination. Serologic evidence of vaccine priming was observed, and all vaccinated groups differed (P < 0.05) from the control at different time points. MD, NDV, and IBD protection results after concomitant double-shot single-needle vaccination were near 85%, 95%, and 100%, respectively. Taken together the results indicate no deleterious effects on the efficacy of the vHVT-IBD vaccine monitored by vaccine intake, serologic and challenge results, and combinations after concomitant live/killed vaccination, suggesting the suitability of its use in hatchery vaccination. All types of injectors used as well as injection techniques, vaccines injected separately or together, gave the same results.     

Electron microscopical studies of the colonization of conidia of Cochliobolus sativus by soil microorganisms

Conidia of Cochlioholus sativus were placed onto natural soil, incubated for periods up to 100 days, recovered and examined by scanning and transmission electron microscopy. Lysis of conidia and their colonization by soil microorganisms were studied.     

Eperythrozoon wenyonii infection in dairy cattle

Approximately 10 of 100 young heifers that had recently delivered their first calf--members of a large Colorado dairy herd--had a syndrome of swollen teats and distal portions of the hind limbs, prefemoral lymphadenopathy, transient fever, rough coat, decreased milk production, and subsequent weight loss and reproductive inefficiency. Acute clinical signs of disease were associated with large numbers of Eperythrozoon wenyonii seen on blood smears, and resolution of signs correlated with reduction or disappearance of the parasite. Other known causes of peripheral edema could not be documented. The parasite was transmitted to 4 of 7 nonlactating dairy cows destined to be culled and a splenectomized calf via IV inoculation of blood from parasitemic heifers, but clinical signs of infection were not induced.     

一个农牧结合生态系统物流模型的建立及持续性分析

建立了人口、玉米连作与奶牛养殖相结合的农牧生态系统物流模型。并运用Ｃ语文编制程序进行预测，经多年实验数据检验效果较好，说明模型有效、可行并具有一定通用性。最后将模型用于哈尔滨玉米带，探索几种不同的农业类型的结构与功能，并进行持续性分析。