PCR-Based Molecular Characterization of Toxocara spp. Using Feces of Stray Cats: A Study from Southwest Iran

Feces of stray cat are potential sources of gastrointestinal parasites and play a crucial role in spreading and transmitting parasite eggs, larvae, and oocysts through contamination of soil, food, or water. In this study, we investigated the prevalence of Toxocara spp. infection in stray cats in Ahvaz city, southwest Iran. Eggs of Toxocara spp. in feces of stray cats were detected by the sucrose flotation method, and identification was conducted by polymerase chain reaction (PCR) and DNA sequencing. Of the 140 fecal samples that were randomly collected from public environments during the months of January to May 2012, 45% were found to harbour Toxocara spp. eggs. The highest prevalence of Toxocara spp. eggs was found in the central area of Ahvaz city (28.6%). T. canis eggs were found in 4 (6.34%) of the 63 positive samples. Stray cats are found in parks, playgrounds, and other public places and may be a potential contamination risk. Identification of Toxocara spp. using molecular methods is sufficiently sensitive to detect low levels of parasites and identify the different Toxocara spp. in feces. The relatively high prevalence of Toxocara spp. infection may continue to increase due to lack of effective environmental hygiene control in Iran. Consequently, there is a need to plan adequate programs to detect, identify, and control this infection as well as stray cats in the region.     

Isolation of Three Xylanase-Producing Strains of Actinomycetes and Their Identification Using Molecular Methods

Eighty-eight actinomycetes were isolated from 20 samples collected from different locations in and around Delhi, India. Among these, 69 isolates were found positive for xylanase production. Of 69 isolates, 3 (SN32, SN77, and SN83) produced >125 IU/ml xylanase. Modern genetic tools were used for revealing the identities of these potent xylanase producers. The selected isolates were categorized under the genus Streptomyces based on their cultural and morphologic characteristics. Genetic diversity among these species of Streptomyces was established based on restriction length fragment polymorphism and random amplified polymorphic DNA analysis. The closest phylogenetic neighbours according to the 16S rRNA gene-sequence data for the three isolates SN32, SN77, and SN83 were Streptomyces cyaneus, S. tendae, and S. caelestis, respectively.     

Identification of Cryptosporidium felis in a Cow by Morphologic and Molecular Methods

Apicomplexan Cryptosporidium parasites infect a wide range of vertebrate hosts. While some species are limited to a single host group, such as Cryptosporidium baileyi, which infects chickens, other species of this genus, such as C. parvum, infect a wide range of mammalian species from mice to humans. During an investigation of Cryptosporidium infection in cattle on a farm in northern Poland, we identified an infection caused by C. felis, in addition to known infections with C. muris and C. parvum. This new infection was identified based on the size of the oocysts (mean size, 4.3 ± 0.4 μm; range, 3.5 to 5.0 μm), as well as by analysis of the molecular sequence of the variable region of the small-subunit rRNA. This finding demonstrates the complex host specificity and circulation in the environment of Cryptosporidium species.     

应用果蝇yellow基因的分子模式分析基因转应作用(英文)

基因转应作用（ｔｒａｎｓｖｅｃｔｉｏｎ）是基因表达的一种方式,这种方式是由等位基因配对及其相互作用所介导的。基因转应作用的现象已在果蝇的多种基因中发现。这种作用可产生正负两种效应。而且,在其它物种中,也逐渐发现了类似的现象。例如,在植物中的基因沉默现象（ｇｅｎｅｓｉｌｅｎｃｉｎｇ）以及在小鼠中的基因转激活作用（ｔｒａｎｓａｃｔｉｖａｔｉｏｎ）等。因此,阐明基因转应作用的机理,将有助于了解基因表达调节及增强子调控活动的分子基础。本文应用果蝇ｙｅｌｏｗ基因为模式来探讨基因转应作用的分子机制。前期研究表明,ｙｅｌｏｗ基因转应作用发生于ｇｙｐｓｙ诱导的ｙ２突变种和ｙｅｌｏｗ亚等位基因（ｙｈ）之间。为了证实是否ｇｙｐｓｙ是基因转应作用所必需的ＤＮＡ元件,我们鉴定了一种新的ｙｅｌｏｗ突变种,称为ｙ２３７４。ｙ２３７４突变是一种基因表达的组织特异性改变,这一改变使ｙ２３７４果蝇在翅和身体部位表皮着色呈突变型。通过遗传分析表明,ｙ２３７４也可与ｙｈ（如ｙ１＃８）产生基因转应作用。ｙ１＃８是一种无效的ｙｅｌｏｗ等位基因,它包含一个启动子和部分编码区序列的缺失。然而,当ｙ２３７４与ｙ１＃８杂交后,其杂交后代的表现型可由ｙ２３７     

Identification and Registration of Maize Genotypes with the Use of Molecular Markers

In plant genetics and breeding, second-generation molecular markers allow detailed characterization of plant genotypes. As shown by PCR analysis of ten SSR loci, 40 genotypes were unique. For every locus, SSR analysis revealed heterozygotes among simple hybrids, which made it possible to identify the parental forms with a high probability of exclusion of nonparental forms. A system was proposed for registration of maize genotypes in the form of genetic formulas reflecting the allelic state of microsatellite loci, in order to catalog, preserve, and effectively employ the existing maize gene pool in breeding.     

Detection of Trichomonas Vaginalis Infection in Chronic Prostatitis/Chronic Pelvic Pain Syndrome Patients by Rapid Immunochromatographic Test

This study aims to evaluate associations between the immunochromatographic rapid test technique and Trichomonas vaginalis (TV) infection in patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) in Taiwan. All patients received post-prostate massage urine (VB3) Trichomonas rapid tests. The demographic characteristics and urogenital symptoms of CP/CPPS were recorded. Routine urinalysis of VB3 was also performed, and laboratory examination results of semen were recorded if available. A total of 29 patients with TV infection and 109 without TV infection were enrolled, which reflected that the prevalence in patients with TV infection was approximately 21%. Patients with TV infection displayed a significantly higher frequency of suprapubic/lower abdominal pain (p = 0.034), semen leukocyte > 5/high-power field (HPF) (p = 0.020), and an inflammatory type (category IIIA) (p = 0.005) than patients without TV infection. A higher prevalence of TV infection was found in patients with category IIIA (47.37%). No significant difference was found in the symptom duration and other clinical symptoms. In conclusion, the high prevalence of TV infection was revealed in CP/CPPS patients using the VB3 rapid Trichomonas test, especially in CP/CPPS patients with category IIIA. Thus, rapid TV testing might be vital for CP/CPPS patients in the hospital.     

中老年阴道炎患者阴道毛滴虫感染状况调查分析

目的:分析中老年阴道炎患者阴道毛滴虫感染情况,为中老年阴道炎的防治提供依据。方法:对2016年9月～2019年9月来我院就诊的中老年阴道炎患者的阴道分泌物进行阴道毛滴虫检查,同时通过问卷调查方法收集患者的病历资料,回收有效问卷200份,分析不同年龄、季节、职业、生活习惯、性生活情况患者的阴道毛滴虫感染状况。结果:中老年阴道炎患者阴道毛滴虫感染率为10.50%,其中45～50岁年龄段阴道炎患者阴道毛滴虫感染率最高,占19.15%,71～80岁年龄段阴道炎患者阴道毛滴虫感染率最低,占2.78%,不同年龄段阴道炎患者阴道毛滴虫感染率比较有统计学差异(P0.05)。春季、夏季阴道毛滴虫感染率显著高于秋季、冬季(P0.05)。牧民、农民阴道炎患者阴道毛滴虫感染率显著高于工人和公职人员的感染率(P0.05),牧民、农民阴道炎患者阴道毛滴虫感染率无明显差异(P0.05)。经常性清洁外阴的阴道炎患者阴道毛滴虫感染率显著低于非经常性清洁外阴的患者(P0.05)。有性生活阴道炎患者阴道毛滴虫感染率显著高于无性生活患者(P0.05),有性生活应用避孕套的阴道炎患者阴道毛滴虫感染率显著低于不应用避孕套患者(P0.05)。结论:中老年阴道炎患者阴道毛滴虫感染率较高,春季、夏季是阴道毛滴虫感染的高峰期,中年患者感染率高于老年患者,农民、牧民、服务人员和非经常性清洁外阴的患者阴道毛滴虫感染率较高,且有性生活没有应用避孕套者感染率更高,应做好以上高危人群的防治工作,以降低阴道毛滴虫的感染率。     

分子标记在鉴别拟船叶藓属和猫尾藓属中的应用

船叶藓科的拟船叶藓属(Dolichom itriopsis)和猫尾藓属植物(Isothecium)非常相似,不易从外部表型上进行属间物种的区分.以尖叶拟船叶藓6个居群、猫尾藓1个居群及疑似尖叶拟船叶藓种1个居群为材料,进行了随机扩增多态性(RAPD)标记分析,结果表明:50个随机引物中有15个引物扩增的产物具有多态性,15个引物共扩增出77条带,其中多态性条带为71条,多态带比率高达92.21%.聚类分析结果表明,疑似种与猫尾藓有较近的亲缘关系,与尖叶拟船叶藓亲缘关系较远.同时测定了疑似尖叶拟船叶藓种的rDNA的ITS全序列,与Genbank已知序列比对结果表明,疑似种与猫尾藓的同源性高达94.74%,而与尖叶拟船叶藓的同源性只有80.64%.进一步支持了RAPD的聚类结果,说明疑似种是猫尾藓而非尖叶拟船叶藓.     

Detection and Enumeration of Vibrio vulnificus in Oysters from Two Estuaries along the Southwest Coast of India, Using Molecular Methods

This study was conducted to understand the seasonal distribution of Vibrio vulnificus in oysters from two estuaries and the effect of environmental factors on the abundance of V. vulnificus in tropical waters. V. vulnificus was detected in 56.6% of the samples tested by colony hybridization with an alkaline phosphatase-labeled oligonucleotide probe (VV-AP), and the counts ranged from <10/g during the summer months to 10³/g in the monsoon season at both sites. The density of V. vulnificus appeared to be controlled more by salinity than by temperature. A nested PCR used in this study detected V. vulnificus in 85% of the samples following 18 h of enrichment in alkaline peptone water.     

利用简并引物克隆黑莓、树莓肌动蛋白基因片段及表达分析

采用同源克隆法,利用设计的一对肌动蛋白(actin)基因简并引物,从黑莓、树莓和悬钩子杂种3个悬钩子植物品种均得到一条743 bp的actin cDNA片段,推测编码247个氨基酸.序列比对发现,克隆的actin基因推导的氨基酸序列和其他植物的肌动蛋白氨基酸序列具有高度保守性,经数据库搜索后将来自黑莓和树莓品种的actin cDNA片段在GenBank中登录,登录号分别为HQ439558和HQ439559.不同来源的植物actin蛋白系统进化树的聚类结果表明,树莓和悬钩子杂种亲缘关系较为密切.actin基因在3个悬钩子植物品种不同组织中的RT-PCR分析结果表明,其在检测的不同组织中均有一定程度的表达量,可能都属于组成型表达的肌动蛋白基因.研究结果为利用actin基因作为内参进一步研究黑莓和树莓其他基因的丰度奠定基础.     

Identification of antigenic proteins in Trichomonas vaginalis

Trichomoniasis is a sexually transmitted disease due to infection with Trichomonas vaginalis, and it can cause serious consequences for women's health. To study the virulence factors of this pathogen, T. vaginalis surface proteins were investigated using polyclonal antibodies specific to the membrane fractions of T. vaginalis. The T. vaginalis expression library was constructed by cloning the cDNA derived from mRNA of T. vaginalis into a phage λ Uni-ZAP XR vector, and then used for immunoscreening with the anti-membrane proteins of T. vaginalis antibodies. The immunoreactive proteins identified included adhesion protein AP65-1, α-actinin, kinesin-associated protein, teneurin, and 2 independent hypothetical proteins. Immunofluorescence assays showed that AP65-1, one of the identified immunogenic clones, is prevalent in the whole body of T. vaginalis. This study led us to identify T. vaginalis proteins which may stimulate immune responses by human cells.     

基于线粒体COI基因序列的壮真蝎与普洱真蝎的分子鉴定

对蝎类物种的传统分类主要依靠形态和行为特征,但由于该类群种间形态特征极为相似,物种的划分和鉴定困难。为弥补传统分类方法的不足,本研究以线粒体细胞色素氧化酶亚基Ⅰ(COI)基因作为分子标记,对形态相似的壮真蝎Euscorpiops validus和普洱真蝎E.puerensis进行分子水平的物种鉴定。采用PCR扩增测序获得壮真蝎与普洱真蝎共24个样本的COI基因部分片段序列(660 bp),进行了遗传距离、系统发育及单倍型网络图分析。结果显示:壮真蝎15个样本中共检测到4个单倍型,单倍型之间的相似度为99.3%~99.8%;普洱真蝎9个样本中共检测到4个单倍型,单倍型之间的相似度为99.6%~99.8%;2种蝎的种间序列相似度为90.1%~90.6%,单倍型间的稳定差异核苷酸位点数为61个。壮真蝎与普洱真蝎种内平均遗传距离分别为0.004 0、0.002 3,种间平均遗传距离为0.103 9,且种间遗传距离为种内的34.6倍。此外,分子系统发育树显示壮真蝎与普洱真蝎的单倍型序列各自聚为2个单系枝,且具有很高的分枝自举值(100%)。单倍型网络图结果也显示壮真蝎与普洱真蝎8个单倍型明显分为2大类群,且壮真蝎的单倍型HAP2与普洱真蝎的单倍型HAP7之间的突变步数高达62步。上述结果不仅进一步确认壮真蝎与普洱真蝎为2个不同的物种,且表明线粒体COI基因可用于开展真蝎属Euscorpiops物种的分子鉴定。     

蚕豆蛋白质亚基分析与特异种质鉴定

采用SDS-PAGE方法,对112份不同基因型蚕豆的清蛋白和球蛋白亚基的差异性进行分析。结果显示:(1)蚕豆清蛋白和球蛋白亚基的有效等位变异分别为1.750 0±0.452 3、1.545 5±0.522 2,多态性比率分别为75.00%、54.55%,清蛋白的亚基遗传多样性指数较球蛋白亚基高。(2)蚕豆清蛋白和球蛋白分别含有在不同基因型蚕豆中构成不同的12和10个亚基,其中清蛋白含有9个基本亚基,116kD、96kD、45kD为清蛋白的3个特异亚基;球蛋白含有8个基本亚基,58kD、35kD为球蛋白的2个特异亚基;研究共鉴定筛选出42个含有清蛋白特异亚基和21个含有球蛋白亚基的种质资源。(3)清蛋白的97kD、63kD基本亚基和球蛋白的97kD、56kD、47kD基本亚基存在一定的缺失现象,共鉴定出19个清蛋白亚基和21个球蛋白亚基缺失的优异种质。研究表明,蚕豆清蛋白和球蛋白亚基构成在不同种质之间具有差异性,除基本亚基外,部分种质还含有特异亚基或缺失亚基。