多重聚合酶链反应筛选方法的研究

本文提出了筛选多重聚合酶链反应的常用方法，根据该法将56个牛微卫星组成的21个多重聚合酶链反应组合，其中14个三微卫星组合，7个二微卫星组合，这些多重聚合酶链反应组合可用于大批量测定微卫星的试验，如基因定位及亲子鉴定等。     

Lectin histochemical investigations of the distal gut of chicks with special emphasis on the follicle-associated epithelium

Carbohydrates on epithelial cell surfaces play an important role as attachment sites for different microorganisms like bacteria, viruses and protozoa. To obtain more information about the distribution of carbohydrates on the luminal surface along the intestine, lectin histochemical studies on different gut segments of chicks of different age groups were carried out using a panel of 13 lectins with specificities for Man, Glc, Gal, GalNAc, GlcNAc or GlcNAc oligosaccharides and Sia. Furthermore, we tried to find out whether previously reported specificities of certain lectins for M cells (membranous or multifold cells) in the bursa of Fabricius (BF) can be observed also on M cells of the intestine. As a result we were able to demonstrate binding of all lectins employed in these studies in all investigated gut segments. In some cases, the application of the same lectin led to varying staining intensities of the same histological structures in different age-groups (e.g. staining of the brush border with WGA, LEA, MAA or Conarva) or different gut segments (e.g. staining of goblet cells with CMA II, LEA and MPA). Hence, terminal carbohydrate residues of glycoconjugates on the intestinal epithelium vary depending on age and organ site. As glycoconjugates can act as attachment sites for microorganisms, these differences in the distribution of sugar residues may be one explanation for the site-specificity of certain pathogens. Furthermore, the binding of lectins to the follicle-associated epithelium (FAE) of the BF differs from that to the FAE of the intestine again stressing the site specificity of lectin binding. Thus, up to now no universal M-cell marker along the chicken intestine exists.     

Prevalence of four enteropathogens in the faeces of young diarrhoeic dairy calves in Switzerland

The prevalences of Cryptosporidium parvum, rotavirus, bovine coronavirus (bcv), and enterotoxigenic Escherichia coli (E coli k99) were determined in diarrhoeic dairy calves aged one to 21 days on 71 dairy farms in western Switzerland during the winter of 2005 to 2006. Faecal samples from 147 untreated calves suffering from acute diarrhoea were analysed by standardised diagnostic methods, and the immunoglobulin status of each calf was evaluated. The prevalences of C parvum, rotavirus, bcv and E coli k99 were 55.0 per cent, 58.7 per cent, 7.8 per cent and 5.5 per cent, respectively. The proportions of herds positive for the respective pathogens among the herds with diarrhoeic calves were 41.7 per cent, 52.1 per cent, 2.1 per cent and 2.1 per cent. The immunoglobulin concentration in the serum of 90.5 per cent of the diarrhoeic calves was below 8 g/l.     

Synthesis of Nitrogen Pools and Fluxes from European Forest Ecosystems

To investigate which ecosystem parameters determine the risk and magnitude of nitrate leaching we compiled data from published and unpublished sources on dissolved inorganic nitrogen (DIN: NO₃ ^-) in throughfall, DIN leaching loss in runoff or seepage water, and other ecosystem characteristics from 139 European forests. Not all data were available for all sites: 126 sites had at least one year's data on DIN inputs and DIN leaching loss; 40-50 sites had some data on soil chemistry and/or vegetation pools of N. DIN inputs in throughfall range between <1 and about 70 kg N ha-1 yr-1 and the losses with seepage or runoff range between <1 and 50 kg N ha-1 yr-1. Retention of N within the ecosystem increases with increasing DIN deposition and increasing proportion of NH₄ ⁺ in deposition. The amount of N in needles and litterfall shows a significant linear relationship with throughfall deposition of DIN, whereas the C:N ratio of the organic (OH) horizon is uncorrelated to the level of throughfall-DIN flux. About 50% of the variability in DIN leaching loss can be explained by the flux of DIN in throughfall. Alternatively, about 60% of the variability in DIN leaching loss can be explained in a two-variable multiple regression combining the C:N ratio of the organic soil and the pH of the mineral soil. The survey data suggest that leaching of DIN from forest ecosystems in Europe is related in part to current DIN deposition and in part to the longer-term internal ecosystem N status as reflected in the chemistry of the humus and acidification status of the soil.     

Influence of pH and soil copper on adsorption of metalaxyl and penconazole by the surface layer of vineyard soils

The upper horizons of old vineyard soils have substantial copper contents due to the traditional use of copper-based fungicides. Total copper levels in eight vineyard soils in the Rías Baixas area of Galicia (northwestern Spain) ranged from 60 to 560 mg kg(-1) (mean +/- SD = 206 +/- 170 mg kg(-1)). The adsorption of the fungicides metalaxyl (pK(a) = 1.41) and penconazole (pK(a) = 2.83) by these soils was determined using fungicide solutions of pH 2.5 and 5.5, and desorption of fungicide adsorbed at pH 5.5 was also determined. In all cases, Freundlich equations were fitted to the data with R (2) > 0.96. Penconazole was adsorbed and retained more strongly than metalaxyl, with K(F) values more than an order of magnitude greater. In the desorption experiments, both fungicides exhibited hysteresis. Soil copper content hardly affected the adsorption of metalaxyl, but K(F) values for adsorption of penconazole increased at a rate of about 0.1 mL(n) (microg of penconazole)(1-n) (microg of Cu)(-1), which is attributed to the formation of Cu(2+)-penconazole complexes with greater affinity for soil colloids than penconazole itself. Because the dependence of K(F) for penconazole adsorption on copper content was the same at both pH values, complex formation appears not to have been affected by the solubilization of 6-17% of soil copper at pH 2.5. A similar copper dependence, or lack of dependence, was observed when 100-1000 mg kg(-1) of copper was added as Cu(NO(3))(2).2H(2)O to the solutions from which the fungicides were adsorbed.     

Changes in red wine soluble polysaccharide composition induced by malolactic fermentation

The polysaccharide content of wine is generally assumed to originate from grapes and yeasts, independent of bacterial metabolism, except for the action of certain spoilage species. This study shows that malolactic fermentation (MLF) significantly modifies the soluble polysaccharide (SP) concentration of various red Bordeaux wines. Wines with the highest initial SP concentration go on to present decreased SP concentration, whereas those with the lowest initial SP concentration rather go on to have a higher SP concentration after MLF. These tendencies were observed whatever the Oenococcus oeni strain (indigenous or starter) used for MLF. Neutral and charged SPs were affected, but to a degree that depended on the microorganisms driving the MLF. The SP modifications were directly linked to bacterial development, because non MLF controls did not present any significant change of SP concentration.     

Survival times of and prognostic indicators for dogs with heart base masses: 25 cases (1986-1999)

OBJECTIVE: To determine survival times of and prognostic indicators for dogs with heart base masses. DESIGN: Retrospective study. ANIMALS: 25 dogs. PROCEDURE: Medical records were reviewed to obtain information regarding signalment; initial complaint; results of physical examination, radiography, electrocardiography, and echocardiography; surgical findings; medical and surgical treatment; outcome; necropsy findings; survival time; and cause of death. RESULTS: Survival time ranged from 0 to 1,096 days (mean, 213 days; median, 57 days). None of the variables examined, including initial complaints and results of physical examination, radiography, electrocardiography, and echocardiography, were associated with survival time. However, dogs that underwent pericardectomy had a significantly longer mean survival time (mean +/- SD, 661 +/- 170 days) than did dogs that were treated medically (129 +/- 51 days). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that dogs with heart base masses that undergo pericardectomy may survive longer than those that receive medical treatment alone.     

Changes in free amino acids and sugars in potatoes due to sulfate fertilization and the effect on acrylamide formation

To examine how sulfur deprivation may affect acrylamide formation in cooked potatoes, three varieties of potato were grown under conditions of either severe sulfur deprivation or an adequate supply of sulfur. In all three varieties sulfur deprivation led to a decrease in acrylamide formation, even though the levels of sugars, which are acrylamide precursors, were higher in tubers of the sulfur-deprived plants. In one variety the concentration of free asparagine, the other precursor for acrylamide, was also higher. There was a very close correlation between the concentration of asparagine in the tubers expressed as a proportion of the total free amino acid pool and the formation of acrylamide upon cooking, whereas sugars were poorly correlated with acrylamide. In potatoes, where concentrations of sugars are usually limiting, competition between asparagine and other amino acids participating in the Maillard reaction may be a key determinant of the amount of acrylamide that is formed during processing.     

日本血吸虫谷胱甘肽S－转移酶小鼠免疫试验

用谷胱甘肽琼脂糖亲和层析柱纯化中国大陆株日本血吸虫成虫谷胱甘肽Ｓ－转移酶（ＧＳＴ）。把纯化的ＧＳＴ结合福氏性剂免疫了二批达ＢＡＬＢ／ｃ小鼠，获得了２９．５８～３２．７１％的减虫率和５２．９４～６８．１３％的粪便减卵率。ＥＬＩＳＡ试验表明免疫小鼠产生了体液免疫应答，免疫印迹试验（ＷｅｓｔｅｒｎＢｌｏｔｔｉｎｇ）显示日本血吸虫水溶性成虫抗原和ＧＳＴ抗原的２６、２８ｋＤ蛋白被免疫小鼠血清所识别。     

Pharmacokinetic indices for cefovecin after single‐dose administration to adult sea otters (Enhydra lutris)

Seven sea otters received a single subcutaneous dose of cefovecin at 8 mg/kg body weight. Plasma samples were collected at predetermined time points and assayed for total cefovecin concentrations using ultra‐performance liquid chromatography and tandem mass spectrometry. The mean (±SD) noncompartmental pharmacokinetic indices were as follows: C_Max (obs) 70.6 ± 14.6 μg/mL, T_{Max (obs)} 2.9 ± 1.5 h, elimination rate constant (k_el) 0.017 ± 0.002/h, elimination half‐life (t_1/2kel) 41.6 ± 4.7 h, area under the plasma concentration‐vs.‐time curve to last sample (AUC_last) 3438.7 ± 437.7 h·μg/mL and AUC extrapolated to infinity (AUC_0→∞) 3447.8 ± 439.0 h·μg/mL. The minimum inhibitory concentrations (MIC) for select isolates were determined and used to suggest possible dosing intervals of 10 days, 5 days, and 2.5 days for gram‐positive, gram‐negative, and Vibrio parahaemolyticus bacterial species, respectively. This study found a single subcutaneous dose of cefovecin sodium in sea otters to be clinically safe and a viable option for long‐acting antimicrobial therapy.