First-trimester prenatal diagnosis of spinal muscular atrophy using microsatellite markers

Twenty-five pregnancies at risk for spinal muscular atrophy I (SMA I) have been monitored by first-trimester prenatal diagnosis. Microsatellite markers were used in all cases to amplify polymorphic regions at the D5S125, D5S435, D5S39, D5S127, and D5S112 loci. All families, including 12 SMA I pedigrees with a deceased index child, were fully informative for DNA analysis. Three fetuses were predicted to be affected and 22 fetuses were predicted to be unaffected. Twenty-two newborns were unaffected by clinical examination at birth. These results support the accuracy of SMA I prenatal diagnosis based on linkage analysis.     

不同TOC/NH4+-N对厌氧氨氧化脱氮效能的影响

采用SBR厌氧氨氧化反应器,研究了不同TOC与NH_4~+-N比值对厌氧氨氧化反应器的脱氮效能的长短期影响.结果表明,在有机物短期影响时,反应器所能承受的最大TOC/NH_4~+-N为1.4,总氮去除速率可达0.26 kg·(m~3·d)~(-1).长期影响下,在TOC/NH_4~+-N小于0.4时,反应器可获得最高脱氮效能,总氮去除率为0.34 kg·(m~3·d)~(-1),TOC/NH_4~+-N大于0.4后,反应器脱氮效能持续降低,并且短期内厌氧氨氧化菌难以迅速恢复活性.利用q PCR(定量PCR)技术对长期影响前后反应器内菌种群落变化做定量分析,结果表明随着有机物的增加,反应器中的ANAMMOX菌数量从2.9×10~(11)copies·mL~(-1)减少至3.15×10~(10)copies·mL~(-1),在TOC/NH_4~+-N大于1.6的环境中,NH_4~+-N未能由厌氧氨氧化菌去除,厌氧氨氧化菌不能表现出生物活性.此时测得反硝化菌数量为3.0×10~9copies·mL~(-1),反应器中的NO_2~--N绝大部分由反硝化去除,虽然反硝化菌数量远少于ANAMMOX菌,但能表现出远超ANAMMOX菌的活性.     

生物除磷系统中聚磷菌检测常用技术

作为生物除磷系统中的主要功能菌,对聚磷菌菌群进行定性、定量分析是深入理解生物除磷系统、提高除磷效率的必然趋势。目前常用于检测聚磷菌的方法主要有生物化学法和分子生物学法,文章主要阐述了荧光原位杂交技术、聚合酶链反应技术、变性梯度凝胶电泳技术以及多技术结合使用的特点及应用情况,并在此基础上提出了改进措施以及聚磷菌检测技术的发展方向。     

The Fate of the Recombinant DNA in Corn During Composting

Abstract

In order to make regulations that safeguard food and the environment, an understanding of the fate of transgenes from genetically modified (GM) plants is of crucial importance. A compost experiment including mature transgenic corn plants and seeds of event Bt 176 (Zea mays L.) was conducted to trace the fate of the transgene cryIA(b) during the period of composting. In bin 1, shredded corn plants including seeds were composted above a layer of cow manure and samples from the corn layer were collected at intervals during a 12-month period. The samples were tested for the transgene persistence and microbial counts and also the compost was monitored for temperature. In bin 2, piles of corn seeds, surrounded by sheep manure and straw, were composted for 12 months. A method combining nested polymerase chain reaction (PCR) and southern hybridization was developed for detection of the transgene in compost. The detection sensitivity was 200 copies of the transgene per gram of dry composted corn material. Composting commenced on day 0, and the transgene was detected in specimens from bin 1 on days 0 and 7 but not on day 14 or thereafter. The transgene in corn seeds was not detectable after 12 months of composting in bin 2. Temperatures in both bins rose to about 50°C within 2 weeks and remained above that temperature for about 3 months, even when the ambient temperature dropped below ?20°C. Extracts from compost were inoculated onto culture plates and then were incubated at 23 to 55°C. Within the first 2 weeks of composting in bin 1, the counts of bacteria incubated at 55°C increased from 3.5 to 7.5 log ₁₀, whereas those incubated at 23°C remained at about 7.5 log ₁₀. The counts of fungi incubated at 45°C increased slightly from 2.5 to 3.1 log₁₀, but those incubated at 23°C decreased from 6.3 to 3.0 log ₁₀. The rapid degradation of the transgene during composting of Bt corn plants suggested that the composting process could be used for safe disposal of transgenic plant wastes.     

Determining heavy metals in spent compact fluorescent lamps (CFLs) and their waste management challenges: Some strategies for improving current conditions

From environmental viewpoint, the most important advantage of compact fluorescent lamps (CFLs) is reduction of green house gas emissions. But their significant disadvantage is disposal of spent lamps because of containing a few milligrams of toxic metals, especially mercury and lead. For a successful implementation of any waste management plan, availability of sufficient and accurate information on quantities and compositions of the generated waste and current management conditions is a fundamental prerequisite. In this study, CFLs were selected among 20 different brands in Iran. Content of heavy metals including mercury, lead, nickel, arsenic and chromium was determined by inductive coupled plasma (ICP). Two cities, Tehran and Tabriz, were selected for assessing the current waste management condition of CFLs. The study found that waste generation amount of CFLs in the country was about 159.80, 183.82 and 153.75 million per year in 2010, 2011 and 2012, respectively. Waste generation rate of CFLs in Iran was determined to be 2.05 per person in 2012. The average amount of mercury, lead, nickel, arsenic and chromium was 0.417, 2.33, 0.064, 0.056 and 0.012 mg per lamp, respectively. Currently, waste of CFLs is disposed by municipal waste stream in waste landfills. For improving the current conditions, we propose by considering the successful experience of extended producer responsibility (EPR) in other electronic waste management. The EPR program with advanced recycling fee (ARF) is implemented for collecting and then recycling CFLs. For encouraging consumers to take the spent CFLs back at the end of the products’ useful life, a proportion of ARF (for example, 50%) can be refunded. On the other hand, the government and Environmental Protection Agency should support and encourage recycling companies of CFLs both technically and financially in the first place.     

污泥臭氧氧化处理过程中活菌抗药基因丰度的消减

臭氧氧化技术是一种广泛应用的污泥减量技术,然而臭氧处理能否对污泥中的抗生素抗药基因进行有效消减还不清楚。采用单叠氮溴化丙锭（propidium monoazide,PMA）预处理结合定量PCR（qPCR）方法对污泥臭氧减量过程中不同臭氧消耗量下活性污泥活菌中的四环素、氨基糖苷和大环内酯3大类共21种抗药基因的变化进行了研究。结果表明：臭氧氧化可以有效消减活性污泥活菌中21种抗药基因的绝对丰度（每毫升污泥的抗药基因拷贝数）,在臭氧消耗量0.31 g·g-1（TSS）情况下抗药基因总量降低了75.44%;尽管绝对丰度下降,污泥臭氧氧化处理过程中大量抗药基因的相对丰度（抗药基因拷贝数与细菌16S rRNA基因拷贝数的比例）逐渐增加,表明臭氧处理后污泥中耐药菌占总活菌比例可能增加,具有一定的抗性传播风险;Ⅰ型整合子是抗药基因水平转移的重要遗传元件,其绝对丰度同样随着臭氧消耗量增加而降低,而相对丰度逐渐上升。     

南京地区污水厂、自来水厂及长江中抗性基因MCR-1和NDM-1的污染特征

近年来,新型抗性基因以其易传播和耐药广等特性,展现出比传统抗性基因更严峻的健康风险,在临床卫生领域受到广泛关注,但目前对其在环境中的行为和风险研究很少.为此,考察了2种有代表性的新型抗性基因MCR-1和NDM-1的污染特征,并借助荧光定量PCR探索了长江下游（南京段）及附近污水厂和自来水厂中MCR-1和NDM-1的分布特征,进而采用RDA（冗余性分析）评价了分布特征受水质指标的影响效果.结果表明:①污水厂进水中MCR-1和NDM-1绝对丰度较高,且随处理流程呈下降趋势,总去除率分别为92.5%和92.7%,但出水中MCR-1和NDM-1绝对丰度仍分别达2.5×108和7.0×106 copies/L.②长江下游（南京段）各采样点MCR-1和NDM-1绝对丰度的范围分别为8.5×107~3.5×109和4.3×105~2.1×107 copies/L,随水流方向呈降低趋势,但在个别采样点出现异常升高的情况,主要受该区域人为污染的影响.③自来水厂处理工艺对MCR-1和NDM-1去除率分别为75.0%和70.6%,但出水中存留的MCR-1和NDM-1绝对丰度分别达1.4×107和6.3×104 copies/L,且MCR-1和NDM-1在排泥水中大量富集.④MCR-1绝对丰度与ρ（COD_Cr）、ρ（NH₃-N）、电导率呈正相关,而NDM-1绝对丰度仅和浊度存在弱相关关系,与其他水质指标无明显相关性.研究显示,污水处理工艺无法有效去除MCR-1和NDM-1,大量抗性基因通过污水厂出水排入长江,同时自来水厂以含有较高绝对丰度抗性基因的长江水作为水源水,最终自来水厂出水中残存的抗性基因可能进入人体,生态健康风险较大.      

聚合酶链式反应(PCR)用于检测环境水体指示菌的研究

对聚合酶链式反应（ｐｏｌｙｍｅｒａｓｅ ｃｈａｉｎ ｒｅａｃｔｉｏｎ，ＰＣＲ）快速检测水体指示菌－大肠菌群（ｔｏｔａｌ ｃｏｌｉｆｏｒｍ）和大肠杆菌（Ｅｓｃｈｅｒｉｃｈｉａ ｃｏｌｉ）进行了研究。采用的两对引物分别扩增了ＬａｃＺ基因的约２６０ｂｐ和ＵｄｉＡ基因的约１５０ｂｐＤＮＡ片段。引物Ⅰ和引物Ⅱ最低能分别扩增１０＾－６μｇ或１０＾－８μｇ基因组ＤＮＡ。整个检测工作在采集水样后５－６ｈ内完成。与     

Effects of elevated CO2 on dynamics of microcystin-producing and non-microcystin-producing strains during Microcystis blooms

In an attempt to elucidate the effects of different CO₂ concentrations (270, 380, and 750 μL/L) on the competition of microcystin-producing (MC-producing) and non-MC-producing Microcystis strains during dense cyanobacteria blooms, an in situ simulation experimentwas conducted in the Meiliang Bay of Lake Taihu in the summer of 2012. The abundance of total Microcystis and MC-producing Microcystis genotypes was quantified based on the 16S rDNA and mcyD gene using real-time PCR. The results showed that atmospheric CO₂ elevation would significantly decrease the pH value and increase the dissolved inorganic carbon (DIC) concentration. Changes in CO₂ concentration did not show significant influence on the abundance of total Microcystis population. However, CO₂ concentrations may be an important factor in determining the subpopulation structure of Microcystis. The enhancement of CO₂ concentrations could largely increase the competitive ability of non-MC-producing over MC-producing Microcystis, resulting in a higher proportion of non-MC-producing subpopulation in treatments using high CO₂ concentrations. Concurrently, MC concentration in water declined when CO₂ concentrations were elevated. Therefore, we concluded that the increase of CO₂ concentrations might decrease potential health risks of MC for human and animals in the future.     

Maternal contamination of amniotic fluid demonstrated by DNA analysis

DNA from 16 sets of samples comprising DNA from uncultured amniotic fluid cells, cultured amniotic fluid cells, fetal tissue, and maternal blood was analysed by the polymerase chain reaction (PCR) with AC-repeat primers. The analysis was performed to investigate the presence of contaminating maternal cells in amniotic fluid which would affect the reliability of DNA studies for prenatal diagnosis. In three sets, maternal contamination of uncultured amniotic fluid cells was detected. In one of the three sets, maternal contamination was present in both uncultured and cultured amniotic fluid cells. The use of amniotic fluid cells as a source of DNA for prenatal diagnosis should be limited to cases where the purity of the DNA can be demonstrated prior to the diagnostic test being performed. This limitation in the use of amniotic fluid DNA also extends to other forms of diagnosis relying on the purity of amniotic fluid samples, particularly the new in situ hybridization methods currently being developed.