Analysis of a Secreted Aspartic Peptidase Disruption Mutant of Glomerella cingulata

Peptidases have been implicated in the pathogenicity of fungi that cause disease in plants. Expression of the secreted aspartic peptidase gene (gcsap), of a Glomerella cingulata isolate pathogenic on apples, is induced during appressorium formation. To determine whether the secreted aspartic peptidase (GcSAP) is essential to pathogenicity, gcsap was disrupted using a vector containing a 637 bp fragment of genomic DNA that encodes the sequence spanning the two active site aspartic acid (Asp) residues. To ensure that the truncated gcsap gene products could not have residual peptidase activity the codons for the active site residues Asp¹¹² and Asp²⁹⁷ were both mutated to histidine residues. Both PCR and Southern analysis confirmed disruption of gcsap. Neither gcsap mRNA nor GcSAP activity was detected in the disruption mutant. Pathogenicity tests on fruit from three apple cultivars showed that GcSAP was not required for pathogenicity. The disruption mutant grew on medium containing protein as the sole source of nitrogen because G. cingulata secretes a previously undetected peptidase(s). A serine peptidase that had a pH optimum between pH 7.0 and 8.0 and a K _m of 0.25 mM for the synthetic substrate succinyl-Ala–Ala–Pro–Phe-p-nitroanilide was identified.     

遗传标记及其在家禽育种中的应用

1 SNP SNP是染色体中遗传变异的最普通形式,如在人体已经检测基因组DNA中每1,000到2,000个单位有1个SNP,在某些区域估计每300个单位有一个SNP.基于SNP的高分辨率,遗传图谱在一些人的染色体上通常没有得到发展.正如其名称所指,SNP是发生在基因(启动子、外显子、或内含子)或基因之间(基因间隔区)的单个碱基改变或核苷酸变异.编码序列中的SNP以同义(不会导致氨基酸改变)或非同义(导致氨基酸改变)来分类.非同义SNP由于对蛋白质表达及最终对表现型的潜在影响而受到重视.相反,同义SNP可能对基因表达有微小的影响(除那些在启动子和其它基因组区域中,对DNA-蛋白质作重要的核苷酸或与RNA稳定性有关的核苷酸).同义和非同义SNP在基因图谱研究中均是很好的遗传标记.     

Pharmacokinetic profiles of the two major active metabolites of metamizole (dipyrone) in cats following three different routes of administration

This study was performed to determine pharmacokinetic profiles of the two active metabolites of the analgesic drug metamizole (dipyrone , MET), 4‐methylaminoantipyrine (MAA), and 4‐aminoantipyrine (AA), after intravenous (i.v., intramuscular (i.m.), and oral (p.o.) administration in cats. Six healthy mixed‐breed cats were administered MET (25 mg/kg) by i.v., i.m., or p.o. routes in a crossover design. Adverse clinical signs, namely salivation and vomiting, were detected in all groups (i.v. 67%, i.m. 34%, and p.o. 15%). The mean maximal plasma concentration of MAA for i.v., i.m., and p.o. administrations was 148.63 ± 106.64, 18.74 ± 4.97, and 20.59 ± 15.29 μg/ml, respectively, with about 7 hr of half‐life in all routes. Among the administration routes, the area under the plasma concentration curve (AUC) value was the lowest after i.m. administration and the AUC_EV/i.v. ratio was higher in p.o. than the i.m. administration without statistical significance. The plasma concentration of AA was detectable up to 24 hr, and the mean plasma concentrations were smaller than MAA. The present results suggest that MET is converted into the active metabolites in cats as in humans. Further pharmacodynamics and safety studies should be performed before any clinical use.     

Effects of oral administration of different dosages of carvacrol essential oils on intestinal barrier function in broilers

Essential oils are widely used in the pharmaceutical, food and cosmetic industries, and many plant essential oils have shown that they have positive effects on broilers nutrition. This experiment was conducted to study the effects of orally administered different dosages of carvacrol essential oils on intestinal barrier function in broiler chickens. A total of eighty 28‐day‐old (1.28 ± 0.15 kg) ROSS 308 broilers were randomly allocated to four groups of 20 replicates each, with one chicken per replicate per cage, and all were fed with the same diet. Four experimental groups were orally administered 0, 200, 300 or 400 μl carvacrol essential oils at 18:00 hr every day during the 2‐week experimental period. As a result of which, the gene expression of the occludin, claudin‐1, claudin‐5, ZO‐1 and ZO‐2 in intestinal mucosa of small intestine (p < 0.05) and the goblet cell content in small intestine epithelium (p < 0.05) were significantly increased; test subjects with 300 or 400 μl carvacrol essential oils reduced the microbial counts of Salmonella spp. and Escherichia coli in the intestines (p < 0.05); Essential oils administration also significantly increased activity of the sucrase (p < 0.05) and lactase (p < 0.05) in intestinal mucosa. In conclusion, the carvacrol essential oils have positive effects on growth performance and intestinal barriers function of broilers; those effects may be related to the dosage, as administration of 300 or 400 μl was more effective than that of 200 μl.     

Using the indicator amino acid oxidation technique to study threonine requirements in horses receiving a predominantly forage diet

Threonine has been reported to be the second limiting amino acid in typical equine diets, but its actual requirement has not been determined in horses. To evaluate amino acid metabolism and requirements, the indicator amino acid oxidation (IAAO) method has been successfully used in other species. The objective of this research was to estimate threonine requirements in mature horses fed timothy hay and concentrate in 4:1 ratio using the IAAO method. Six Thoroughbred mares (579.9 ± 46.7 kg) received each of 6 levels of threonine intake, 41, 51, 61, 70, 80 and 89 mg/kg BW/day, in a randomly determined order. Each study period was 7‐day long, and on day 6, blood samples were collected before and 90 min after feeding to measure amino acid concentrations using HPLC. On day 7, horses underwent IAAO procedures, which included a 2‐hr primed, constant intravenous infusion of [¹³C]sodium bicarbonate to measure total CO₂ production and a 4‐hr primed, constant oral administration of [1‐¹³C]phenylalanine to estimate phenylalanine oxidation to CO₂. Blood and breath samples were collected to measure blood [¹³C]phenylalanine, using GC‐MS analysis and breath ¹³CO₂ enrichment, using an infrared isotope analyser. Increasing threonine intake levels did not affect plasma phenylalanine oxidation by the ANOVA test (p > 0.05) but resulted in a linear decrease in phenylalanine oxidation (p = 0.04) without a breakpoint by the orthogonal linear contrast. This study is the first attempt to evaluate threonine requirements in horses by the IAAO method; however, threonine requirements are still unknown in mature horses at this time.     

Equine foetal gender determination in mid‐ to late gestational mares: A practical inquiry

In recent years, the interest in equine foetal gender determination (FGD) during gestation increased remarkably. Ultrasonographic FGD can be performed in two different periods during gestation. The earliest examination can take place at a gestational age of 60–70 days, whereby the genital tubercle is used to differentiate between male and female foeti. The time window of the second approach is wider (120–210 days), and there are more characteristics to take into consideration. In this article, the feasibility and accuracy of ultrasonographic FGD in mid‐ to late gestation are evaluated. One hundred twenty‐one mares from different breeds with a pregnancy stage between 120 and 270 days were examined once, using B‐scale ultrasonography (Esaote MyLab™ClassC). None of the mares were sedated nor shaved, and the procedure was completed within 15 min. Diagnosis was firstly based on the gonads. The final judgement was made based on all visible foetal reproductive organs. In three cases with a pregnancy stage beyond 257 days, FGD was not possible. All of the examined mares in which a FGD could be performed gave birth to a healthy foal. In 98% of the examinations (116/118), the diagnosis was correctly made. In both cases of misdiagnosis, only one characteristic was seen during the procedure and wrongly interpreted. Beyond 210 days of pregnancy, the extremities can preclude a good visualization of the inguinal region. In conclusion, equine FGD in mid‐ to late gestation is an accessible and accurate technique, although a good ultrasound device is a prerequisite and experience and expertise is necessary.     

山东省青岛市畜禽屠宰管理现状

为了解山东省青岛市畜禽屠宰管理发展现状,优化管理政策措施,开展了畜禽屠宰管理现状调查。调查发现:青岛市肉食消费以猪肉为主,牛、羊、禽、兔肉为副;全市现有生猪定点屠宰企业20家,年屠宰量约220万头;家禽屠宰企业4家,年屠宰量2亿只;牛羊屠宰企业6家,年屠宰量2万头。通过对青岛市畜禽屠宰企业监管体系和追溯体系建设,以及无害化处理等内容进行调研分析,指出当前存在代宰模式隐患多、监管责任难厘清、规范管理待完善等问题,并由此提出严格代宰管理,探索"两检合一"措施,改善产品追溯体系,完善行业管理规定,引导健康消费及加大资金投入等发展建议。     

美国动物卫生保护制度框架浅析

基于对美国《动物卫生保护法》的研究,本文概述了其立法宗旨,动物疫病预防、突发疫情应急处置、国内流行病控制消灭等制度框架,以及生产经营者主体责任、调运监管制度、动物卫生实验室体系、兽医认证计划、监督执法制度、经费预算方式等保障措施,以期为完善我国立法体系提供借鉴。     

农药市场常见违法行为分析与建议

本文对四川省近5年立案查处的农药市场违法案件进行整理,对案例反映出的农药市场常见违法行为进行归类分析,并提出了相应的监管建议,试图给农药市场监管工作提供参考。     

荞麦抗旱性研究进展

荞麦是一种珍贵的药食同源作物,随着人们对健康饮食的逐渐重视,荞麦及其加工产品在人们生活中的地位越发重要。中国拥有丰富的荞麦资源,多种植在干旱或半干旱的冷凉高原山区及边远山区。干旱严重影响荞麦的产量和品质,提高荞麦抗旱性在荞麦生产中起着重要的作用。为此,从干旱对荞麦形态特征、抗旱性的生理生化基础、干旱对荞麦产量及品质的影响等方面阐述了荞麦抗旱性的研究进展,并对荞麦抗旱分子机理方面的研究进行了展望。