不同蚯蚓采样方法对比研究

蚯蚓种类组成和丰度变化是反映农业管理措施对土壤质量影响的重要指标。为研究利用芥末悬液等方法进行蚯蚓样品采集的有效性和准确性,寻求合理有效的蚯蚓种群特征调查方法,在我国东北玉米连作黑土上分别运用手拣法、福尔马林溶液驱赶法、芥末+水悬液及芥末+乙酸悬液驱赶法,以及驱虫剂与手拣法相结合的方法进行蚯蚓样品采集,将上述7种方法测得的蚯蚓丰度、生物量及年龄组成信息进行了系统分析。结果表明,福尔马林溶液驱赶法不结合手拣法使用时测得的蚯蚓丰度及生物量远远低于其他方法,尤其幼年蚯蚓比例偏低,并不能反映真实的蚯蚓种群特征。使用芥末悬液做驱虫剂有效性明显提高,尤其芥末+乙酸悬液,测得的蚯蚓丰度比手拣法高22.3%。福尔马林溶液结合手拣法后有效性大大提高,测得的蚯蚓丰度和生物量分别是其单独使用时的19.1和9.3倍,但相对其他驱虫剂方法仍高估了成年蚯蚓的比例;芥末+水悬液和芥末+乙酸悬液方法结合手拣法后收集到蚯蚓的数量分别提高了67.8%和89.1%,平均个体重量分别是原来的1.8和1.3倍,说明芥末悬液不会杀死小个体的幼年蚯蚓,但少部分大个体蚯蚓及洞穴不与地表相接的蚯蚓可能并不能被其驱赶至地表,结合手拣法后可以弥补这个缺陷。芥末+乙酸悬液结合手拣法收集到的成年及幼年蚯蚓数量和生物量均显著高于其他方法(P<0.05),既能有效地驱赶小个体和幼年蚯蚓,又可以收集到洞穴不与地表相接的土栖型蚯蚓,因此可以作为真实有效地反映蚯蚓种群特征的采样方法之一。     

长白山地区不同采集土样方式筛菌效率比较

目的对随机采样和田字格定点采样两种取样方式筛菌效率进行比较分析。方法利用随机采样和定点采样两种不同方式,采集了长白山地区的土样,取来自不同植物根际的土样,制成菌悬液后煮沸,涂布无氮平板以分离固氮芽胞杆菌。对分离的菌株进行菌落16S rDNA PCR,从中筛选鉴别巨大芽胞杆菌。结果定点采集土样分离出无晶体芽胞菌和有晶体芽胞菌分别是261株和76株,随机采集土样分离出无晶体芽胞菌和有晶体芽胞菌分别是279株和94株。经16S rDNA法鉴定分离巨大芽胞杆菌共19株。结论随机采样的取土方式与定点采样的取土方式差异度小,两种采集土样的方式在筛菌效率上并未表现出明显差异。     

肺螨标本的采集保存和制作

本文报告了当前用碱性消化液消化、分离痰螨的方法,并且对现有的痰液消化法,和痰液收集法进行了比较和分析。结论为24小时痰液收集法和应用改良消化液(与前人分离痰螨的方法相比较)能提高检出率(P<0.01)。     

东湖原生动物群落结构变化与水质差异的相关研究

东湖自 60年代以来 ,富营养化程度不断加剧 ,由于各湖区接纳污水的程度不同 ,使不同水域的水质状况呈现明显的差别。本研究对东湖污染程度不同的水域设立 6个采样站 ,进行原生动物的采集及鉴定 ,目的是研究水污染造成的原生动物群落结构的变化 ,同时也是对各种结构和功能参数在水生态系统污染的监测和评价中的作用进行比较。1　方法1 .1　采样及镜检在东湖共设 6个采样站 (图 1 ) ,采用ＰＦＵ法[1]收集原生动物。于 1 995年 1月1 5日在各站挂上空白ＰＦＵ ,2月 1 5日取回。用ＰＦＵ挤出液进行活体定性观察 ,在高 ( 4 0× )、中 ( 2 0× )、…     

收集挥发性样品的吸附剂法和固相微萃取法

本文介绍挥发性信息化学物质的两种收集方法——吸附剂收集法和固相微萃取收集法,以植物挥发性化学物质收集为例为例,详述了两种方法的原理、步骤、注意事项和实例,最后比较了两种收集方法的优缺点,供相关工作者使用时参考。     

植物挥发性气体(VOCs)研究进展

植物挥发性气体(VOCs)在植物一植食性昆虫-天敌三级营养关系、植物间信息传递及适应性改变上都发挥着重要作用.植物释放VOCs具特异性、系统性、时序性与节律性等特点,VOCs主要在寄主选择行为、产卵行为、求偶行为、引来昆虫夭敌干涉等方面影响植食性昆虫.VOCs-介导的植物间信息传递作用包括4个过程:"释放者"植物合成及释放气体、气体在空气中的运输、气体在植物表面的吸附及"接收者"植株对气体信号的感知.收集VOCs的方法主要有吸附-溶剂洗脱法和吸附-热脱附法.     

基于硅胶管吸附萃取的热脱附-气相色谱-质谱联用法测定真菌的挥发性有机化合物

【背景】真菌代谢产生的挥发性有机化合物(Volatile organic compounds,VOCs)具有重要的生物学功能。高通量的VOCs测定方法对于VOCs产生菌的筛选非常重要。【目的】建立一种简单可靠、基于硅胶管吸附萃取(Siliconetubesorptiveextraction,STSE)的热脱附-气相色谱-质谱(Thermal desorption-gas chromatography-mass spectrometry,TD-GC-MS)联用法测定真菌培养物VOCs的高通量分析技术。【方法】选择代表性的包括酯、酮、醇、醛、烯等21种VOCs作为对照品建立标准曲线;将对照品滴加于PDA培养基,基于STSE进行TD-GC-MS分析,测定其专属性;将木霉菌T552接种于PDA培养基,基于STSE进行TD-GC-MS分析,测定方法的重复性;在以上基础上,用上述试验条件对6株木霉菌产生的VOCs进行筛选。【结果】在全扫描和提取离子条件下,21种对照品分别在1.0-10.0μg/mL、0.1-10.0μg/mL范围内线性关系良好。基于STSE的TD-GC-MS分析发现,滴加在PDA培养基上的对照品分离良好,且PDA培养基平板中的其它物质不干扰VOCs的测定,表明该方法专属性良好;对PDA培养的木霉菌T552进行的VOCs重复测定鉴定的5种VOCs相对标准偏差均小于10%,表明该方法重复性良好。用建立的基于STSE的TD-GC-MS方法从6株供试木霉菌中鉴定出包括酸、醇、酯、烯等共37种VOCs。【结论】建立了一种简单可靠、基于STSE的TD-GC-MS分析测定真菌培养物VOCs的高通量分析技术,可用于真菌VOCs的分析测定。     

葡萄果实香气成分的厌氧萃取研究

采用厌氧液-液萃取对葡萄果实中的香气成分进行了萃取,用气相色谱-质谱联用技术对其进行成分分析,并与葡萄果实常规萃取方法得到的香气成分进行比较。结果显示,采用厌氧萃取方法得到的萃取物含有的香气成分较多,一次性萃取可分析得到39种香气成分,且以内标物2-辛醇表示的香气成分含量明显高于常规萃取方法;香气成分归属的化学类别更多,比分液漏斗萃取方法多出2类香气成分。结果表明,厌氧萃取所得果实香气成分更真实,更典型,为葡萄果实香气成分的分析提供了一种有效的提取方法。     

超声微波协同萃取(UMAE)番茄中番茄红素的研究

研究了超声微波协同萃取番茄中的番茄红素。以乙酸乙酯为萃取溶剂并以不同的提取时间、微波功率和液固比作为参数进行实验,并进行了工艺优化。在提取时间367s、微波功率98W和液固比10．6：1的条件下实际提取率可达到97．4％。实验结果表明：UMAE法是一种能够快速有效的提取番茄红素的方法。     

提高丁醇萃取发酵耦联生产改良型生物柴油过程中萃余液回用效率的研究

利用少量廉价的萃取剂一正辛醇（正辛醇／萃余液体积比0．2：1）对萃余液中的丁醇进行萃取,萃余液中56％的残余丁醇可被回收,总丁醇得率提高了38％。萃余液经活性炭（3％,w／v）处理后,在100％回用拌料条件下一批次发酵能够正常进行。萃取发酵条件下,反复全回用萃余液5次,萃取发酵性能可以保持稳定。为丁醇萃取发酵耦联生产改良型生物柴油过程中萃余液的全回用提供了一种新型、低成本和绿色环保的方法。     

The Approach to Sample Acquisition and Its Impact on the Derived Human Fecal Microbiome and VOC Metabolome

Recent studies have illustrated the importance of the microbiota in maintaining a healthy state, as well as promoting disease states. The intestinal microbiota exerts its effects primarily through its metabolites, and metabolomics investigations have begun to evaluate the diagnostic and health implications of volatile organic compounds (VOCs) isolated from human feces, enabled by specialized sampling methods such as headspace solid-phase microextraction (hSPME). The approach to stool sample collection is an important consideration that could potentially introduce bias and affect the outcome of a fecal metagenomic and metabolomic investigation. To address this concern, a comparison of endoscopically collected (in vivo) and home collected (ex vivo) fecal samples was performed, revealing slight variability in the derived microbiomes. In contrast, the VOC metabolomes differ widely between the home collected and endoscopy collected samples. Additionally, as the VOC extraction profile is hyperbolic, with short extraction durations more vulnerable to variation than extractions continued to equilibrium, a second goal of our investigation was to ascertain if hSPME-based fecal metabolomics studies might be biased by the extraction duration employed. As anticipated, prolonged extraction (18 hours) results in the identification of considerably more metabolites than short (20 minute) extractions. A comparison of the metabolomes reveals several analytes deemed unique to a cohort with the 20 minute extraction, but found common to both cohorts when the VOC extraction was performed for 18 hours. Moreover, numerous analytes perceived to have significant fold change with a 20 minute extraction were found insignificant in fold change with the prolonged extraction, underscoring the potential for bias associated with a 20 minute hSPME.     

The Amazonas‐trap: a new method for sampling plant‐inhabiting arthropod communities in tropical forest understory

Methods to quantify plant‐insect interactions in tropical forests may miss many important arthropods and can be time consuming and uneven in capture efficiency. We describe the Amazonas‐trap, a new method that rapidly envelops the target plant for sampling arthropods. We evaluated the efficiency of the Amazonas‐trap by comparing it with two commonly used sampling methods to collect arthropods from plants: the beating tray and manual collection. Samples were collected in 10 permanent plots, in the Ducke forest reserve, Manaus (Amazonas, Brazil). In each plot we sampled 18 plant individuals of Protium sp. (Burseraceae): six by a beating tray, six by manual collection, and six using the Amazonas‐trap. All insects were identified to the family level and those belonging to the order Hymenoptera were identified to the species and morphospecies level. The new method sampled more insect families and more Hymenoptera species than tree beating and manual collection. Of the 75 total families collected, 20 were sampled exclusively by the Amazonas‐trap, seven were only collected with a beating tray, and seven were sampled exclusively with manual collecting. A similar pattern was found for abundance: Amazonas‐trap sampled more individuals, followed by the beating tray and manual collection. Small and winged arthropods were more abundant in Amazonas‐trap, explaining the highest richness of Hymenoptera and insect families sampled with this method. The new method sampled more spiders, wood‐fungi feeders, sap suckers, omnivorous, parasitoids, and insect predators than the other methods, but was equally effective in sampling leaf‐feeders and ants. Amazonas‐trap was more time consuming in the field, but for all diversity parameters evaluated, the new method showed better performance for collecting invertebrates on plants.     

北京市珍珠泉乡不同昆虫采集方法采集效果对比分析

为了探究科研调查中不同昆虫采集方法的采集效率差异,在北京珍珠泉乡不同的生境类型中,采用样线法、马氏网法、灯诱法、陷阱法(糖醋液)4种采集方法进行昆虫采集,按生境类型对不同采集方法采集的昆虫进行统计分析.在实验中共采集昆虫3996头,隶属12目87科,其中在昆虫种类数量上,鳞翅目最多,其次是半翅目和鞘翅目;在数量上,半翅目昆虫数量最多,依次是双翅目和鳞翅目.样线法共采集到昆虫61科72种、灯诱法共采集到45科95种、马氏网法共采集到39科41种、陷阱法共采集到14科25种.在调查中,不同方法在不同生境采集昆虫种类数多存在显著差异(P<0.05).不同的采集方式间除马氏网和样线法采集昆虫的种类相似度为中等不相似(q=0.436),其他方法间均为极不相似.采用不同的采集方式可以采集到更多的昆虫种类.马氏网法是最优选的采集方式,马氏网采集昆虫最方便,节省人力;样线法采集的效果最稳定;灯诱法的采集效果好,但是应用较为复杂;糖醋液陷阱法采集昆虫效率对比其他三种方法,既不能采集到大量昆虫也没有采集到特定种昆虫,如无特别需求可以考虑减少或者放弃使用.     

Impaction onto a Glass Slide or Agar versus Impingement into a Liquid for the Collection and Recovery of Airborne Microorganisms

To study impaction versus impingement for the collection and recovery of viable airborne microorganisms, three new bioaerosol samplers have been designed and built. They differ from each other by the medium onto which the bioaerosol particles are collected (glass, agar, and liquid) but have the same inlet and collection geometries and the same sampling flow rate. The bioaerosol concentrations recorded by three different collection techniques have been compared with each other: impaction onto a glass slide, impaction onto an agar medium, and impingement into a liquid. It was found that the particle collection efficiency of agar slide impaction depends on the concentration of agar in the collection medium and on the sampling time, when samples are collected on a nonmoving agar slide. Impingement into a liquid showed anomalous behavior with respect to the sampling flow rate. Optimal sampling conditions in which all three new samplers exhibit the same overall sampling efficiency for nonbiological particles have been established. Inlet and collection efficiencies of about 100% have been achieved for all three devices at a sampling flow rate of 10 liters/min. The new agar slide impactor and the new impinger were then used to study the biological factors affecting the overall sampling efficiency. Laboratory experiments on the total recovery of a typical environmental microorganism, Pseudomonas fluorescens ATCC 13525, showed that both sampling methods, impaction and impingement, provided essentially the same total recovery when relatively nonstressed microorganisms were sampled under optimal sampling conditions. Comparison tests of the newly developed bioaerosol samplers with those commercially available showed that the incorporation of our research findings into the design of the new samplers yields better performance data than data from currently available samplers.     

Establishing arthropod community composition using metabarcoding: Surprising inconsistencies between soil samples and preservative ethanol and homogenate from Malaise trap catches

DNA metabarcoding allows the analysis of insect communities faster and more efficiently than ever before. However, metabarcoding can be conducted through several approaches, and the consistency of results across methods has rarely been studied. We compare the results obtained by DNA metabarcoding of the same communities using two different markers – COI and 16S – and three different sampling methods: (a) homogenized Malaise trap samples (homogenate), (b) preservative ethanol from the same samples, and (c) soil samples. Our results indicate that COI and 16S offer partly complementary information on Malaise trap samples, with each marker detecting a significant number of species not detected by the other. Different sampling methods offer highly divergent estimates of community composition. The community recovered from preservative ethanol of Malaise trap samples is significantly different from that recovered from homogenate. Small and weakly sclerotized insects tend to be overrepresented in ethanol while strong and large taxa are overrepresented in homogenate. For soil samples, highly degenerate COI primers pick up large amounts of nontarget DNA and only 16S provides adequate analyses of insect diversity. However, even with 16S, very little overlap in molecular operational taxonomic unit (MOTU) content was found between the trap and the soil samples. Our results demonstrate that none of the tested sampling approaches is satisfactory on its own. For instance, DNA extraction from preservative ethanol is not a valid replacement for destructive bulk extraction but a complement. In future metabarcoding studies, both should ideally be used together to achieve comprehensive representation of the target community.     

A rapid method for tissue collection and high-throughput isolation of genomic DNA from mature trees

Collection of tissue and subsequent isolation of genomic DNA from mature tree species often proves difficult. DNA extraction from needles, leaves, or buds is recommended in many protocols. Collecting these tissues from mature trees generally requires the use of firearms or climbing if sampling is to be nondestructive. As a result, sample collection is a major expense of many tree-based projects. Tree (and plant) tissues generally contain large amounts of polysaccharides and phenolic compounds that are difficult to separate from DNA. Many methods aim to overcom these problems, with most involving extraction in buffers containing the nonionic detergent cetyltrimethyl-ammonium bromide (CTAB), followed by numerous steps to clean contaminants from the DNA, using organic solvents and differential salt precipitation. These steps are time-consuming, such that isolation of DNA becomes the bottleneck in many molecular studies. This paper presents a new, efficient, cambium collection method for tree species and a DNA extraction protocol based on that of Doyle and Doyle (1987), with follow-up purification using the Wizard nuclei lysis and protein precipitation solutions (Promega). Results show a significant improvement in yield and DNA purity compared with other published methods, with consistently high yields of pure genomic DNA and high sample throughput. The relatively low cost per extraction, no requirement for use of liquid nitrogen, no requirement for freezer storage, and long-term sample stability after collection are important additional benefits.     

Comparison of pitfall traps and litter bags for sampling ground-dwelling arthropods

Abstract:  For their simplicity and effectiveness, pitfall traps have become a standard technique to measure the activity and relative abundance of ground-dwelling arthropods. Permeable screen or mesh bags filled with plant material, referred to as litter bags, have also recently been employed as a complementary sampling technique for epigeal taxa. The anticipated need for increased field research on arthropod populations, particularly in transgenic crops with insecticidal properties, suggests that a relative assessment of both sampling methods could contribute to the design of future studies. Comparisons among pitfall traps, and litter bags placed above- or below-ground indicated that above-ground litter bags most frequently succeeded in collecting certain groups of arthropods associated with moisture and sheltered areas, including centipedes (Chilopoda) and beetle larvae (Carabidae, Staphylinidae). Conversely, pitfall traps most often captured taxa considered active at ground level, such as adult carabids or harvestmen (Opiliones). For taxa collected in >40% of all three trap types, bootstrap confidence intervals for the coefficient of variation (CV; used to assess precision or sampling efficiency) showed that above-ground litter bags were significantly more precise than pitfall traps for sampling elongate springtails (Collembola) and adult rove beetles (Staphylinidae), but only during the first year of sampling. While below-ground litter bags often appeared similar to one or both of the other trap types, in no case were below-ground litter bags best based on frequency of collection or CV. Though differences were not always consistent between years, results suggest that the additional effort required to sample using litter bags may be justified for the collection of some ground-dwelling taxa.     

Propylene glycol: a promising preservative for insects,comparable to ethanol,from trapping to DNA analysis

Recent advances in DNA analysis allow us to identify an unprecedented number of insect samples collected by mass sampling techniques such as insect traps. In these circumstances, a preservative that can be applied from trap to storage is necessary to prevent degradation of DNA before analysis and save on the cost of labor for collecting insects from traps. Propylene glycol has a prominent feature as a trap solution. We aimed to examine the DNA preservability of 98% propylene glycol at 2 weeks and more than 6 months after initial collection in comparison with 99.5% ethanol, which is commonly used for storage of specimens for genetic analysis. We compared amplification performance of PCR targeting a specific region of the mitochondrial cytochrome c oxidase subunit I (COI) gene in the orders Hymenoptera, Diptera, and Coleoptera using two extraction methods varying in extraction efficiency. Even after 6 months, more than 75% of samples were recognized to have succeeded in PCR amplification irrespective of preservatives by the extraction method with higher extraction efficiency. It suggested that mitochondrial DNA was preserved in both solutions. However, dim bands in the electrophoreses of PCR products increased with time in extracts by another method with lower extraction efficiency. In Diptera and Coleoptera, the rate of dim bands increased more rapidly for ethanol-preserved than for propylene glycol-preserved specimens, indicating higher DNA preservability of propylene glycol over time for these taxa. On the other hand, in Hymenoptera, the preservatives did not affect PCR amplification performance. Considering its safer characteristics and high DNA preservability in a wide range of taxa, propylene glycol can be a promising solution from trapping of insects to storage for genetic analysis.     

Larval densities and a life-table for the gypsy moth, Lymantria dispar, estimated using the head-capsule collection method

ABSTRACT. 1. In order to estimate the absolute larval density in each stage of a larval population of the gypsy moth, Lymantria dispar L., in a deciduous forest in northern Japan, the head-capsule collection method was used. An estimate by this method was compared with estimates based on two kinds of frass collection methods.
2. Twenty-one traps made of cloth were put in a study plot. Each trap was of 24.5 cm diameter. Larval head-capsules falling into the traps were collected and sorted by hand. On the first sampling occasion, the population was also estimated using the frass-collection method.
3. Larval numbers estimated by the head-capsule collection method were almost identical to estimates by the two frass methods. Larval numbers entering the four larval instar were successfully estimated by the head-capsule collection method, and an age-specific life table was established using the resultant estimates.     

A factorial design experiment as a pilot study for noninvasive genetic sampling

Noninvasive genetic sampling has increasingly been used in ecological and conservation studies during the last decade. A major part of the noninvasive genetic literature is dedicated to the search for optimal protocols, by comparing different methods of collection, preservation and extraction of DNA from noninvasive materials. However, the lack of quantitative comparisons among these studies and the possibility that different methods are optimal for different systems make it difficult to decide which protocol to use. Moreover, most studies that have compared different methods focused on a single factor – collection, preservation or extraction – while there could be interactions between these factors. We designed a factorial experiment, as a pilot study, aimed at exploring the effect of several collection, preservation and extraction methods, and the interactions between them, on the quality and amplification success of DNA obtained from Asiatic wild ass (Equus hemionus) faeces in Israel. The amplification success rates of one mitochondrial DNA and four microsatellite markers differed substantially as a function of collection, preservation and extraction methods and their interactions. The most efficient combination for our system integrated the use of swabs as a collection method with preservation at ?20 °C and with the Qiagen DNA Stool Kit with modifications as the DNA extraction method. The significant interaction found between the collection, preservation methods and the extraction methods reinforces the importance of conducting a factorial design experiment, rather than examining each factor separately, as a pilot study before initiating a full‐scale noninvasive research project.