Development and evaluation of a quantitative PCR assay for detection of Hepatozoon sp

With the aim to improve current molecular diagnostic techniques of Hepatozoon sp. in carnivore mammals, we developed a quantitative PCR (qPCR) assay with SYBR Green I^®. The method, consisting of amplification of a 235 bp fragment of the 18S rRNA gene, is able to detect at least 0.1 fg of parasite DNA. Reproducible quantitative results were obtained over a range of 0.1 ng–0.1 fg of Hepatozoon sp. DNA. To assess the performance of the qPCR assay, DNA samples from dogs (140) and cats (50) were tested with either standard PCR or qPCR. Positive samples were always confirmed by partial sequencing of the 18S rRNA gene. Quantitative PCR was 15.8% more sensitive than standard PCR to detect H. canis in dogs. In cats, no infections were detected by standard PCR, compared to two positives by qPCR (which were infected by H. canis as shown by sequencing).     

Biologic and genetic comparison of Toxoplasma gondii isolates in free-range chickens from the northern Pará state and the southern state Rio Grande do Sul, Brazil revealed highly diverse and distinct parasite populations

The prevalence of Toxoplasma gondii in 84 free-range chickens (34 from the northern Pará state, and 50 from Rio Grande do Sul, the southern state) from Brazil, South America was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT), and found in 39 (46.4%) of 84 chickens with titers of 1:10 in one, 1:20 in two, 1:40 in four, 1:80 in seven, 1:160 in five, 1:320 in six, 1:640 in eight and > or =1:1280 in six. Hearts and brains of 45 chickens with titers of 1:20 or less were pooled and fed to two T. gondii-free cats. Hearts and brains of 39 chickens with titers of 1:10 or higher were bioassayed in mice. Feces of cats were examined for oocysts. One cat fed tissues from 31 chickens with titers of less than 1:10 from Rio Grande do Sul shed T. gondii oocysts. T. gondii was isolated by bioassay in mice from 33 chickens with MAT titers of 1:20 or higher. All infected mice from 10 isolates died of toxoplasmosis. All 34 isolates (15 from Pará, 19 from Rio Grande do Sul) were genotyped using 11 genetic markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, a new SAG2 and Apico. Eleven genotypes were revealed for Pará isolates and seven genotypes for Rio Grande do Sul. No genotype was shared between the two geographical locations. These data suggest that T. gondii isolates are highly diverse and genetically distinct between the two different regions in Brazil that are 3500 km apart.     

纤维素分解微生物复合菌剂降解固态物料特性研究

为提高纤维素类碳源物质降解能力,通过试验驯化得到可降解玉米秸秆微生物复合菌剂。将复合菌剂添加到以纤维素类物质（滤纸、打印纸、秸秆、报纸）作为培养基碳源物质的液体培养基中,分析菌剂反应前后培养基中碳源物质减重率。结果表明,滤纸、打印纸、秸秆、报纸减重率分别达到93.7%、83.2%、75.4%、61.3%。玉米秸秆横截面扫描电镜观察表明,复合菌剂处理前后,细胞结构变得疏松,产生裂缝和空洞,有效证实驯化微生物复合菌剂有助于降解纤维素碳源物质能力。为分离复合菌剂中的单菌株,利用功能菌培养机上产生的透明圈及颜色变化,分离出降解蛋白质、脂肪、纤维素及产酸菌株,利用16S rRNA序列分析,鉴定出4个菌属8种菌株,分别为Chryseobacterium indologenes Dan121、Chryseobacterium indologenes Dan191、Acinetobacter bereziniae Zhi71、Acinetobacter bereziniae Zhi77、Acinetobacter bereziniae Zhi111、Lactococcus garviae S36、Bacillus cereus M7、Bacillus cereus MP310。     

玉米对有机氯农药污染土壤的修复潜力(英文)

[目的]确定玉米对土壤中老化滴滴涕和六六六的累积、迁移和转化能力。[方法]温室盆栽试验。取均匀掺入梯度浓度滴滴涕和六六六农药的科研土壤装盆,收集持续盆栽过程中不同时段的土壤和植物样品进行残留测试。[结果]所选择的玉米植株在所设计的土壤污染浓度范围内具有宽域的滴滴涕和六氯环己烷清除能力,其茎秆、根系的生物浓缩系数分别介于0.004至0.027和0.036到0.097之间,但去除率均低于0.1,滴滴涕和六六六梯度处理间无显著差异。滴滴涕可通过被动吸附和主动吸收被玉米植体积累,且由还原性脱卤化介导,p,p’-DDT向p,p’-DDE转化。[结论]玉米对土壤有机氯农药具有清除修复能力,且有机氯农药的对玉米脂肪的亲和力是影响其在植物体内的吸收和转运的主要因素之一。     

Food safety in raw milk production: risk factors associated to bacterial DNA contamination

While human illness from milkborne pathogens may be linked to contamination of the product after pasteurization or improper pasteurization, such diseases are usually associated with consumption of raw milk or its by-products. Molecular biology tools were applied to investigate contamination by Listeria monocytogenes, Salmonella spp., some pathogenic strains of Escherichia coli, and Campylobacter jejuni in 548 raw milk samples from 125 dairy farms established in two regions from southern Brazil. Moreover, 15 variables were evaluated for their association with raw milk contamination levels, and the risk factors were determined by multiple regression analysis. Salmonella spp. were more frequently detected, followed by pathogenic E. coli. There was difference in contamination index between the regions, in which risk factors such as temporary cattle confinement, low milk production, low milking machine cleaning frequency, and milk storage area without tile walls were identified. The risk factors were specific to each region studied. Nevertheless, the data can be used to improve milk quality of dairy farms/herds with similar management practices.     

Genetic analysis of a possible case of canine X-linked ectodermal dysplasia

In the present report, we describe targeted next-generation sequencing of the EDA gene of a male poodle with a clinical and histopathological diagnosis of X-linked hypohidrotic ectodermal dysplasia. The result was compared with the reference sequence and with the result of the sequencing of a normal dog's EDA gene. No point variant, small deletion or insertion were found in the exons and splice sites, but a transition and a transversion were found in the intron 6 and 3′ UTR, respectively. The cause of the dysplasia of the affected dog in this study is neither a point variant nor a small deletion or insertion in the exons and splice sites of the EDA gene. Therefore, patients with phenotype of XLHED may have other types of variants in the EDA gene or variants in other genes of the EDA signalling pathway.     

紫花苜蓿铝激活苹果酸转运蛋白基因的克隆与亚细胞定位

为揭示紫花苜蓿耐铝机理,根据已知的紫花苜蓿(Medicago sativa L)与铝胁迫相关的EST序列,运用RACE技术获得了紫花苜蓿铝激活苹果酸转运蛋白基因的全长cDNA,命名为MsALMT1,这一全长cDNA为1 638 bp,包含一个1 344 bp的最大开放阅读框,编码448个氨基酸,BLAST比对发现,Ms...     

不同干扰和尺度下荒漠啮齿动物群落格局

2002~2004年,在内蒙古阿拉善荒漠区选择4种不同干扰生境,在3个空间尺度上对啮齿动物群落格局进行了研究。结果表明,在不同干扰和尺度下,荒漠啮齿动物群落格局具有明显差异。随着尺度的增大,不同干扰生境种类组成明显增加,优势种组成格局明显变化,开垦干扰下子午沙鼠在3个空间尺度上都为优势种;轮牧干扰下子午沙鼠只是在小尺度上占有优势,而小毛足鼠在2个较大空间尺度上始终为优势种;过牧干扰下同样子午沙鼠只是在小尺度上占有优势,而三趾跳鼠在2个较大空间尺度上始终为优势种;禁牧干扰下子午沙鼠和黑线仓鼠的优势地位在1hm2和10hm22个空间尺度上交替出现,但在40hm2空间尺度则同时成为优势种。在荒漠栖息地破碎化过程中,啮齿动物群落格局在不同空间尺度上受到了明显的影响。三趾跳鼠在群落中数量升高是荒漠生境开始破碎化的表现之一。     

Effects of Monomer Structure on Their Organization and Polymerization in a Smectic Liquid Crystal

Photopolymerizable diacrylate monomers dissolved in fluid-layer smectic A and smectic C liquid crystal (LC) hosts exhibited significant spatial segregation and orientation that depend strongly on monomer structure. Small, flexible monomers such as 1,6-hexanediol diacrylate (HDDA) oriented parallel to the smectic layers and intercalated, whereas rod-shaped mesogen-like monomers such as 1,4-di-(4-(6-acryloyloxyhexyloxy)benzoyloxy)-2-methylbenzene (C6M) oriented normal to the smectic layers and collected within them. Such spatial segregation caused by the smectic layering dramatically enhanced photopolymerization rates; for HDDA, termination rates were reduced, whereas for C6M, both the termination and propagation rates were increased. These polymerization precursor structures suggest novel materials-design paradigms for gel LCs and nanophase-separated polymer systems.     

山楂PCR-SSCP反应体系与反应条件的优化

[目的]优化山楂PCR-SSCP反应体系与反应条件.[方法]以山楂为研究材料,采用改良的CTAB法分步提取叶绿体DNA和核DNA.从8对候选引物中分别筛选适合对叶绿体DNA和核DNA进行PCR扩增的引物,同时进行PCR-SSCP反应体系和反应条件的优化.琼脂糖凝胶电泳检测PCR扩增产物,非变性聚丙烯酰胺凝胶电泳检测变性后的PCR扩增产物.[结果]筛选出ropB、ropL、rpoC1、ITS2和psbA-tmH五对引物适用于山楂的PCR-SSCP分析.电泳时,上样缓冲液(不含甘油)与PCR产物等量混合,98℃碱(1％NaOH)变性15min,采用6％的聚丙烯酰胺凝胶(交联度29∶1),电泳缓冲液0.5×TBE,在4℃恒温和200 V恒压的条件下,电泳3～4 h,可得到较好的山楂PCR-SSCP电泳图谱.[结论]建立了山楂PCR-SSCP最优反应体系与反应条件,为山楂SSCP分析奠定基础.