Aspects of Pitcher Morphology and Spectral Characteristics of Six BorneanNepenthesPitcher Plant Species: Implications for Prey Capture

Pitcher plants of the genusNepenthesattract and trap invertebrateprey using nectar-secreting pitchers. Pitcher morphology andspectral reflectance characteristics were investigated for sixNepenthesspeciesfrom northwest Borneo (N. albomarginata, N. ampullaria, N. bicalcarata,N. gracilis, N. mirabilisvar.echinostomaandN. rafflesiana).Morphological measurements focused on the size of the pitcherrim (or peristome, the site of the major nectaries) in relationto pitcher length. The results show considerable interspecificvariation in morphology. Spectral reflectance measurements quantifiedthe degree of colour contrast between the peristome and pitcherbody, from ultraviolet (UV) to red wavelengths. The contrastmaxima for each species were compared with insect visual sensitivitymaxima. The six species showed a wide range of reflectance patterns,with pitchers ofN. rafflesianapossessing the greatest degreeof ‘fit’ between contrast maxima and insect sensitivitymaxima, in the UV, blue and green regions of the spectrum. Basedon the morphological and reflectance analyses, we hypothesizedthat pitchers ofN. rafflesianawould be more attractive to anthophilous(flower-visiting) invertebrates than the sympatricN. gracilis.Analysis of prey contents generally supported the hypothesis,suggesting possible interspecific resource partitioning. Morphologicaland spectral characteristics of the other species are discussedin relation to published studies on prey capture by those species.Copyright1999 Annals of Botany Company Borneo, carnivory, colour,Nepenthesspp., pitcher plants, prey attraction.     

Pitcher geometry facilitates extrinsically powered ‘springboard trapping' in carnivorous Nepenthes gracilis pitcher plants

Carnivorous pitcher plants capture insects in cup-shaped leaves that function as motionless pitfall traps. Nepenthes gracilis evolved a unique ‘springboard'' trapping mechanism that exploits the impact energy of falling raindrops to actuate a fast pivoting motion of the canopy-like pitcher lid. We superimposed multiple computed micro-tomography images of the same pitcher to reveal distinct deformation patterns in lid-trapping N. gracilis and closely related pitfall-trapping N. rafflesiana. We found prominent differences between downward and upward lid displacement in N. gracilis only. Downward displacement was characterized by bending in two distinct deformation zones whist upward displacement was accomplished by evenly distributed straightening of the entire upper rear section of the pitcher. This suggests an anisotropic impact response, which may help to maximize initial jerk forces for prey capture, as well as the subsequent damping of the oscillation. Our results point to a key role of pitcher geometry for effective ‘springboard'' trapping in N. gracilis.     

Stable Isotope Ecology of Extant Tapirs from the Americas

Understanding the ecology of modern and ancient forests can help clarify the evolution of forest dwelling mammals. It is first necessary, however, to elucidate the source and extent of stable isotope variation in forest taxa. Tapirs are of particular interest because they are model organisms for identifying forest environments due to their highly conservative diet and habitat preferences. Here, stable carbon and oxygen isotopic compositions of extant tapirs are quantified to test hypotheses regarding ontogenetic diet shifts, stable isotope variation at the population level, and relationships between stable isotopes and climatic variables. A population of extant tapirs (Tapirus bairdii) demonstrates low δ¹³C variation (～2–3‰) and increased δ¹³C values in late erupting teeth, indicating that juveniles consume ¹³C deplete milk and/or browse in denser forests. Disparate δ¹⁸O values of late erupting teeth are instead reflecting seasonal variation. Extant tapir (T. bairdii, Tapirus pinchaque, Tapirus terrestris) δ¹⁸O values are constrained by climatic and geographic variables. Most notably, δ¹⁸O values of T. bairdii decrease with decreasing precipitation frequency. Tapirus terrestris is typically present in areas with greater precipitation than T. bairdii and δ¹⁸O values instead positively correlate with δ¹³C values. These data indicate that tapirs in wetter areas are getting a larger proportion of their water from leaves experiencing less evaporation in denser canopies, while T. bairdii is interpreted to increase its consumption of water via drinking when present in drier areas. An understanding of extant tapir stable isotope ecology improves ecological interpretations of these elusive mammals both today and in the past. Abstract in Spanish is available at http://www.blackwell‐synergy.com/loi/btp .     

Stable Isotope Analysis in Primatology: A Critical Review

Stable isotope analysis has become an important tool in ecology over the last 25 years. A wealth of ecological information is stored in animal tissues in the relative abundances of the stable isotopes of several elements, particularly carbon and nitrogen, because these isotopes navigate through ecological processes in predictable ways. Stable carbon and nitrogen isotopes have been measured in most primate taxonomic groups and have yielded information about dietary content, dietary variability, and habitat use. Stable isotopes have recently proven useful for addressing more fine‐grained questions about niche dynamics and anthropogenic effects on feeding ecology. Here, we discuss stable carbon and nitrogen isotope systematics and critically review the published stable carbon and nitrogen isotope data for modern primates with a focus on the problems and prospects for future stable isotope applications in primatology. Am. J. Primatol. 74:969‐989, 2012. © 2012 Wiley Periodicals, Inc.     

Stable Isotope Ratios and Forensic Analysis of Microorganisms

In the aftermath of the anthrax letters of 2001, researchers have been exploring various analytical signatures for the purpose of characterizing the production environment of microorganisms. One such signature is stable isotope ratios, which in heterotrophs, are a function of nutrient and water sources. Here we discuss the use of stable isotope ratios in microbial forensics, using as a database the carbon, nitrogen, oxygen, and hydrogen stable isotope ratios of 247 separate cultures of Bacillus subtilis 6051 spores produced on a total of 32 different culture media. In the context of using stable isotope ratios as a signature for sample matching, we present an analysis of variations between individual samples, between cultures produced in tandem, and between cultures produced in the same medium but at different times. Additionally, we correlate the stable isotope ratios of carbon, nitrogen, oxygen, and hydrogen for growth medium nutrients or water with those of spores and show examples of how these relationships can be used to exclude nutrient or water samples as possible growth substrates for specific cultures.     

Aberrant Water Homeostasis Detected by Stable Isotope Analysis

While isotopes are frequently used as tracers in investigations of disease physiology (i.e., ¹⁴C labeled glucose), few studies have examined the impact that disease, and disease-related alterations in metabolism, may have on stable isotope ratios at natural abundance levels. The isotopic composition of body water is heavily influenced by water metabolism and dietary patterns and may provide a platform for disease detection. By utilizing a model of streptozotocin (STZ)-induced diabetes as an index case of aberrant water homeostasis, we demonstrate that untreated diabetes mellitus results in distinct combinations, or signatures, of the hydrogen (δ²H) and oxygen (δ¹⁸O) isotope ratios in body water. Additionally, we show that the δ²H and δ¹⁸O values of body water are correlated with increased water flux, suggesting altered blood osmolality, due to hyperglycemia, as the mechanism behind this correlation. Further, we present a mathematical model describing the impact of water flux on the isotopic composition of body water and compare model predicted values with actual values. These data highlight the importance of factors such as water flux and energy expenditure on predictive models of body water and additionally provide a framework for using naturally occurring stable isotope ratios to monitor diseases that impact water homeostasis.     

Purple Pitcher Plant (Sarracenia rosea) Dieback and Partial Community Disassembly following Experimental Storm Surge in a Coastal Pitcher Plant Bog

Sea-level rise and frequent intense hurricanes associated with climate change will result in recurrent flooding of inland systems such as Gulf Coastal pitcher plant bogs by storm surges. These surges can transport salt water and sediment to freshwater bogs, greatly affecting their biological integrity. Purple pitcher plants (Sarracenia rosea) are Gulf Coast pitcher plant bog inhabitants that could be at a disadvantage under this scenario because their pitcher morphology may leave them prone to collection of saline water and sediment after a surge. We investigated the effects of storm surge water salinity and sediment type on S. rosea vitality, plant community structure, and bog soil-water conductivity. Plots (containing ≥1 ramet of S. rosea) were experimentally flooded with fresh or saline water crossed with one of three sediment types (local, foreign, or no sediment). There were no treatment effects on soil-water conductivity; nevertheless, direct exposure to saline water resulted in significantly lower S. rosea cover until the following season when a prescribed fire and regional drought contributed to the decline of all the S. rosea to near zero percent cover. There were also significant differences in plant community structure between treatments over time, reflecting how numerous species increased in abundance and a few species decreased in abundance. However, in contrast to S. rosea, most of the other species in the community appeared resilient to the effects of storm surge. Thus, although the community may be somewhat affected by storm surge, those few species that are particularly sensitive to the storm surge disturbance will likely drop out of the community and be replaced by more resilient species. Depending on the longevity of these biological legacies, Gulf Coastal pitcher plant bogs may be incapable of fully recovering if they become exposed to storm surge more frequently due to climate change.     

Isolation of Viable Multicellular Glands from Tissue of the Carnivorous Plant,Nepenthes

Many plants possess specialized structures that are involved in the production and secretion of specific low molecular weight compounds and proteins. These structures are almost always localized on plant surfaces. Among them are nectaries or glandular trichomes. The secreted compounds are often employed in interactions with the biotic environment, for example as attractants for pollinators or deterrents against herbivores.Glands that are unique in several aspects can be found in carnivorous plants. In so-called pitcher plants of the genus Nepenthes, bifunctional glands inside the pitfall-trap on the one hand secrete the digestive fluid, including all enzymes necessary for prey digestion, and on the other hand take-up the released nutrients. Thus, these glands represent an ideal, specialized tissue predestinated to study the underlying molecular, biochemical, and physiological mechanisms of protein secretion and nutrient uptake in plants. Moreover, generally the biosynthesis of secondary compounds produced by many plants equipped with glandular structures could be investigated directly in glands.In order to work on such specialized structures, they need to be isolated efficiently, fast, metabolically active, and without contamination with other tissues. Therefore, a mechanical micropreparation technique was developed and applied for studies on Nepenthes digestion fluid. Here, a protocol is presented that was used to successfully prepare single bifunctional glands from Nepenthes traps, based on a mechanized microsampling platform. The glands could be isolated and directly used further for gene expression analysis by PCR techniques after preparation of RNA.     

Stable Isotope Probing Analysis of Interactions between Ammonia Oxidizers

The response of natural microbial communities to environmental change can be assessed by determining DNA- or RNA-targeted changes in relative abundance of 16S rRNA gene sequences by using fingerprinting techniques such as denaturing gradient gel electrophoresis (DNA-DGGE and RNA-DGGE, respectively) or by stable isotope probing (SIP) of 16S rRNA genes following incubation with a ¹³C-labeled substrate (DNA-SIP-DGGE). The sensitivities of these three approaches were compared during batch growth of communities containing two or three Nitrosospira pure or enriched cultures with different tolerances to a high ammonia concentration. Cultures were supplied with low, intermediate, or high initial ammonia concentrations and with ¹³C-labeled carbon dioxide. DNA-SIP-DGGE provided the most direct evidence for growth and was the most sensitive, with changes in DGGE profiles evident before changes in DNA- and RNA-DGGE profiles and before detectable increases in nitrite and nitrate production. RNA-DGGE provided intermediate sensitivity. In addition, the three molecular methods were used to follow growth of individual strains within communities. In general, changes in relative activities of individual strains within communities could be predicted from monoculture growth characteristics. Ammonia-tolerant Nitrosospira cluster 3b strains dominated mixed communities at all ammonia concentrations, and ammonia-sensitive strains were outcompeted at an intermediate ammonia concentration. However, coexistence of ammonia-tolerant and ammonia-sensitive strains occurred at the lowest ammonia concentration, and, under some conditions, strains inhibited at high ammonia in monoculture were active at high ammonia in mixed cultures, where they coexisted with ammonia-tolerant strains. The results therefore demonstrate the sensitivity of SIP for detection of activity of organisms with relatively low yield and low activity and its ability to follow changes in the structure of interacting microbial communities.Molecular characterization of natural microbial communities has demonstrated the existence of novel high-level taxonomic groups with no cultured representatives and with significant diversity within phylogenetic and functional groups already established through analysis of organisms in laboratory culture. Autotrophic ammonia-oxidizing bacteria (AOB) exemplify the latter situation. Their low growth rates and the limited number of readily measured phenotypic characteristics available for identification of these organisms necessitate the use of molecular techniques for characterization of their diversity in natural environments. Phylogenetic analysis of 16S rRNA gene sequences places the majority of cultivated autotrophic bacterial ammonia oxidizers in a monophyletic group within the Betaproteobacteria (8, 26). Amplification and phylogenetic analysis of 16S rRNA gene sequences from enrichment cultures of ammonia oxidizers and sequences of environmental clones (31) suggest the existence of novel groups with no cultivated representative and considerable diversity within those represented by pure cultures.Increased awareness of microbial diversity has raised questions regarding links between species diversity and functional diversity, functional redundancy, and the influence of environmental conditions on the activities of representatives of different phylotypes. For ammonia-oxidizing bacteria, relationships exist between broad phylogenetic groups and the environments from which laboratory isolates were obtained, which are linked, in some cases, to differences in physiological characteristics (11). There is also evidence of links between the relative abundance of different ammonia oxidizer groups and environmental conditions (1, 13, 14, 18, 21, 23, 34), suggesting selection for organisms with particular physiological characteristics. In one study (36), a combination of molecular and physiological studies has demonstrated links between species diversity, functional diversity, and soil nitrification kinetics. However, for ammonia oxidizers and other groups, there is little direct evidence about which strains within diverse communities are active under particular conditions or the extent of competition for substrates.Stable isotope probing (SIP) (24, 27) of nucleic acids provides direct evidence of which members of mixed communities are active. This involves addition of substrates labeled with a stable isotope (most commonly ¹³C), extraction of nucleic acids, separation of ¹²C- and ¹³C-labeled nucleic acids by density gradient centrifugation, and subsequent molecular analysis. Sequences amplified from ¹³C-labeled DNA or RNA are derived from organisms actively assimilating the substrate. This approach has been used to identify organisms that utilize methane or methanol (4, 19), organic compounds (15, 20), or CO₂ (6, 9) in microcosms and those that assimilate plant root exudates in the field (28). SIP therefore links phylogeny to ecosystem function and has identified established and novel groups by utilizing labeled compounds in complex soil communities. The technique also enables in situ physiological studies and investigation of interactions between organisms in mixed cultures belonging to the same functional group. For autotrophic betaproteobacterial ammonia oxidizers, amplification of 16S rRNA genes from ¹³C-labeled DNA during incubation with [¹³C]CO₂ has the potential for discriminating which strains are active under specific conditions. Assessment of the discriminatory ability of this approach in complex natural environments requires studies under controlled and well-characterized conditions. The first aim of this study was, therefore, to assess the ability of SIP to discriminate activities of different members of simple mixed communities in comparison with direct measurement of product concentration and DNA- and RNA-denaturing gradient gel electrophoresis (DGGE). The second was to determine whether the activities of members of mixed communities of ammonia-oxidizing bacteria, in particular, their ability to grow at high ammonia concentrations, could be predicted from their physiological characteristics in monoculture. Of particular interest was whether strains with low ammonia tolerance are competitive at low ammonia concentrations. Mixed cultures were assembled from pure culture representatives of Nitrosospira clusters 0, 3a, and 3b (26, 36), which are frequently found in soil environments, and from enrichment cultures containing representatives of these clusters with heterotrophic contaminants. Other criteria for choice of community members were similarities in specific growth rate and cultivation conditions to enable meaningful competition experiments.     

Prey Specialization by Cougars on Feral Horses in a Desert Environment

Natural controls on the distribution, abundance, or growth rates of exotic species are a desirable mode of intervention because of lower costs compared to anthropogenic controls and greater social acceptance. In the Great Basin, cougars (Puma concolor) are the most widely distributed carnivore capable of killing large ungulate prey. Populations of feral horses (Equus ferus) are widely distributed throughout the Great Basin and can grow at rates up to 20%/year. Although cougars exhibit distributional overlap with horses, it has been assumed that predation is minimal because of differences in habitat use and body-size limitations. To evaluate this hypothesis, we monitored the diets of 21 global positioning system (GPS)-collared cougars in the western Great Basin (5 males, 8 females) and eastern Sierra Nevada (2 males, 6 females) from 2009–2012. We investigated 1,310 potential kill sites and located prey remains of 820 predation events. We compared prey composition and kill rates of cougars inhabiting the Sierra Nevada and Great Basin, and among male and female cougars across seasons. We used generalized linear mixed models (GLMMs) to examine the effects of prey availability and habitat characteristics on the probability of predation on horses by cougars. Mule deer (Odocoileus hemionus) comprised 91% of prey items killed on the Sierra Nevada reference site but only comprised 29% of prey items in the Great Basin study area. Average annual kill rates for deer differed between the Sierra Nevada ( = 0.85 deer/week, range = 0.44–1.3) and Great Basin ( = 0.21 deer/week, range = 0.00–0.43). Diets of cougars in the Great Basin were composed predominantly of horses (59.6%, n = 460 prey items; 13 individuals). Ten cougars regularly consumed horses, and horses were the most abundant prey in the diet of 8 additional individuals in the Great Basin. Cougars on average killed 0.38 horses/week in the Great Basin (range=0.00–0.94 horses/week). Differences in predation on horses between the sexes of cougars were striking; Great Basin females incorporated more horses across all age classes year-round, whereas male cougars tended to exploit neonatal young during spring and summer before switching to deer during winter. Within GLMM models, the probability of predation on horses compared to other prey species increased with elevation, horse density, and decreasing density of mule deer on the landscape, and was more likely to occur in sagebrush (Artemesia spp.) than in pinyon (Pinus monophylla)–juniper (Juniperus osteosperma) forests. Behavior of individual cougars accounted for more than a third of the variation explained by our top models predicting predation on horses in the Great Basin. At landscape scales, cougar predation is unlikely to limit the growth of feral horse populations. In the Great Basin ecosystem, however, cougars of both sexes successfully preyed on horses of all age classes. Moreover, some reproductive, female cougars were almost entirely dependent on feral horses year-round. Taken together, our data suggest that cougars may be an effective predator of feral horses, and that some of our previous assumptions about this relationship should be reevaluated and integrated into management and planning. © 2021 The Wildlife Society.     

鸟类组织稳定同位素分析样品的预处理方法

稳定同位素分析样品在进行同位素质谱分析之前需要适当的处理,然而如何处理尚未形成统一的标准。本文结合鸟类组织稳定同位素分析样品的测试流程,介绍了鸟类学研究领域稳定同位素技术应用中涉及的分析样品解冻清洗、干燥、分离纯化、研磨和储存等前处理方法,并对不同组织常用的处理方法及分析样品测定中有待解决的问题展开了讨论。旨在抛砖引玉,为利用稳定同位素技术开展鸟类学方面的研究提供科学参考和技术支持。     

Quantifying Inter-Laboratory Variability in Stable Isotope Analysis of Ancient Skeletal Remains

Over the past forty years, stable isotope analysis of bone (and tooth) collagen and hydroxyapatite has become a mainstay of archaeological and paleoanthropological reconstructions of paleodiet and paleoenvironment. Despite this method''s frequent use across anthropological subdisciplines (and beyond), the present work represents the first attempt at gauging the effects of inter-laboratory variability engendered by differences in a) sample preparation, and b) analysis (instrumentation, working standards, and data calibration). Replicate analyses of a ¹⁴C-dated ancient human bone by twenty-one archaeological and paleoecological stable isotope laboratories revealed significant inter-laboratory isotopic variation for both collagen and carbonate. For bone collagen, we found a sizeable range of 1.8‰ for δ¹³C_col and 1.9‰ for δ¹⁵N_col among laboratories, but an interpretatively insignificant average pairwise difference of 0.2‰ and 0.4‰ for δ¹³C_col and δ¹⁵N_col respectively. For bone hydroxyapatite the observed range increased to a troublingly large 3.5‰ for δ¹³C_ap and 6.7‰ for δ¹⁸O_ap, with average pairwise differences of 0.6‰ for δ¹³C_ap and a disquieting 2.0‰ for δ¹⁸O_ap. In order to assess the effects of preparation versus analysis on isotopic variability among laboratories, a subset of the samples prepared by the participating laboratories were analyzed a second time on the same instrument. Based on this duplicate analysis, it was determined that roughly half of the isotopic variability among laboratories could be attributed to differences in sample preparation, with the other half resulting from differences in analysis (instrumentation, working standards, and data calibration). These findings have serious implications for choices made in the preparation and extraction of target biomolecules, the comparison of results obtained from different laboratories, and the interpretation of small differences in bone collagen and hydroxyapatite isotope values. To address the issues arising from inter-laboratory comparisons, we devise a novel measure we term the Minimum Meaningful Difference (MMD), and demonstrate its application.     

内蒙古巴彦淖尔市纳林套海汉墓出土人骨的稳定同位素分析

稳定同位素分析技术近年来发展为复原古代民族食物结构、社会经济模式的有效手段。本文应用该技术对内蒙古自治区纳林套海汉代墓葬出土人骨中的C、N同位素比值进行了测定。结果表明, 纳林套海汉代居民日常饮食结构中保持着非常高的动物性食物摄入, 植物类食物的摄入中以C4类植物为主。结合其他相关研究结果, 我们认为西汉王朝通过实施移民屯垦和属国等政策, 大力推动了河套地区的农业和畜牧业生产, 改变了河套地区原有的经济模式和人们的饮食结构。本文的研究结果可以为复原汉代北部边疆的经济模式研究提供有益的线索和证据。     

Thermal Sensitivity as a Specialization for Prey Capture and Feeding in Snakes

Behavior can be regarded as a result of various processes ofdecision based on the information provided by the sensory organs.In this review the role of the so-called additional heat sense,next to vision, smell and mechanoreception is discussed withrespect to the feeding behavior of snakes. The hierarchy ofthe sensory information in various phases of the feeding behaviordiffers between snakes possessing heat receptors (e.g., speciesof the Crotalinae and Pythoninae) and those without (e.g., speciesof the Viperinae and Colubrinae). Probably depending on theinfluence of ecological demands, visual or chemical cues arethe main information in the behavioral phases before the strikeHowever, in situations with little visual input, e.g., in darkness,rodents' burrows, etc., hunting behavior is guided in the firstplace by radiation of warm objects in Crotalus, Python and Trimeresurusflavoviridis, and by substrate vibrations in Vipera aspis, Pituophismelanoleucus and Boa constrictor. I suggest that in the sensoryhierarchy, heat information functionally replaces the mechanicalinformation which is utilized by snakes without pit organs.Poststrike behavior on the other hand is mainly guided by chemicalcues in all snakes.     

猪笼草消化液中蛋白酶的活性初探

为了进一步分析猪笼草瓶状体内消化液中蛋白质的性质,本研究在不同的温度和pH条件下测定消化液中蛋白酶的活性,并且比较了3种不同沉淀方法对消化液蛋白进行的浓缩,通过SDS-聚丙烯酰胺电泳对消化液蛋白作了初步分离.结果表明,猪笼草消化液中蛋白酶的最适酶活温度为50℃,且稳定性最高;当pH为5时,该蛋白酶活性出现峰值,采用氯仿-正丁醇法(5:1)沉淀浓缩猪笼草消化液蛋白样品效果最佳.聚丙烯酰胺电泳结果表明,消化液至少包括三种蛋白组分,分子量分别为24.3kD、35.1 kD和61.4kD,且35.1 kD条带具有抗胰蛋白酶消化活性.本研究为综合开发利用猪笼草野生资源提供理论依据.     

高寒草甸生态系统食物链结构分析--来自稳定性碳同位素的证据

稳定性同位素技术应用于生态系统物质流动和食物链营养关系的研究方法 ,是基于生物体内天然存在的同位素比值与它们食物密切相关这一原理建立起来的。即将生物体内的稳定性同位素比值 (如δ1 3C)作为一自然标记 ,根据物种间该值的相对差异 ,追踪生态系统中的主要物质 (如碳源 )的来源和物质流动。于 2 0 0 2年 4～8月测定了高寒草甸生态系统主要生物群落中物种的稳定性碳同位素比值 (δ1 3C) ,依据得到的系统的富集因子( 1 0 5± 0 4 5 )‰ ,分析并确定了所测物种间的取食与被取食关系。结果发现 :高寒草甸生态系统由 5条主要的食物链构成 ,其中 1条为“植物→小型哺乳类→食肉兽 /猛禽”的三节点食物链 ,2条为“植物→牲畜和植物→植食性鸟类”的二节点食物链 ,2条分别为“植物→昆虫→雀形目鸟类→猛禽 /食肉兽”和“植物→昆虫→两栖类→猛禽 /食肉兽”的四节点食物链 ;系统食物链的最大长度为 3 5 3 ,与系统的最大节点数相近。表明稳定性碳同位素可以作为分析高寒草甸生态系统食物网和食物链结构以及食物链长度的有效代理 (proxy)。     

Stable Isotope Probing Analysis of the Diversity and Activity of Methanotrophic Bacteria in Soils from the Canadian High Arctic

The melting of permafrost and its potential impact on CH₄ emissions are major concerns in the context of global warming. Methanotrophic bacteria have the capacity to mitigate CH₄ emissions from melting permafrost. Here, we used quantitative PCR (qPCR), stable isotope probing (SIP) of DNA, denaturing gradient gel electrophoresis (DGGE) fingerprinting, and sequencing of the 16S rRNA and pmoA genes to study the activity and diversity of methanotrophic bacteria in active-layer soils from Ellesmere Island in the Canadian high Arctic. Results showed that most of the soils had the capacity to oxidize CH₄ at 4°C and at room temperature (RT), but the oxidation rates were greater at RT than at 4°C and were significantly enhanced by nutrient amendment. The DGGE banding patterns associated with active methanotrophic bacterial populations were also different depending on the temperature of incubation and the addition of nutrients. Sequencing of the 16S rRNA and pmoA genes indicated a low diversity of the active methanotrophic bacteria, with all methanotroph 16S rRNA and pmoA gene sequences being related to type I methanotrophs from Methylobacter and Methylosarcina. The dominance of type I methanotrophs over type II methanotrophs in the native soil samples was confirmed by qPCR of the 16S rRNA gene with primers specific for these two groups of bacteria. The 16S rRNA and pmoA gene sequences related to those of Methylobacter tundripaludum were found in all soils, regardless of the incubation conditions, and they might therefore play a role in CH₄ degradation in situ. This work is providing new information supporting the potential importance of Methylobacter spp. in Arctic soils found in previous studies and contributes to the limited body of knowledge on methanotrophic activity and diversity in this extreme environment.Permafrost regions occupy approximately 22% of the exposed land area of the Northern Hemisphere (63). In the past 100 years, the average temperatures in the arctic regions have increased at almost twice the average global rate (25). The melting of permafrost is one of the most important impacts of global warming on these high-latitude environments, and theoretical modeling suggests that as much as 90% of the permafrost could thaw by the end of the 21st century (29). While it has been generally reported that 15% of the total soil organic carbon is stocked in permafrost (42), a recent estimate indicates that it contains as much as 50% of the global belowground organic carbon pool (53). Carbon stocked in permafrost is now regarded as one of the most important carbon-climate feedbacks because of the size of the carbon pool and the intensity of climate change at high latitudes (46, 47). The presence of these large amounts of carbon in permafrost is raising serious concerns whether melting permafrost, and the resulting increase in microbial activity, might be a source of extensive emissions of the greenhouse gases carbon dioxide and methane (CH₄) to the atmosphere. The actual emissions of CH₄ from soils of high latitudes have been estimated to represent about 25% of the emissions from natural sources (19). Methane, which is 25 times more potent than carbon dioxide as a greenhouse gas (25), is produced by methanogenic archaea under anaerobic conditions. These microorganisms are known to inhabit permafrost environments (44, 49), and their capacity to produce methane at cold temperatures has been reported (20, 35, 44, 56). Their methanogenic activity is expected to increase as permafrost soil temperature increases (20). Moreover, large amounts of methane are stocked as methane hydrates in permafrost at an average depth of several hundred meters (33). Methane is also found in permafrost layers near the surface and could potentially be liberated to the atmosphere as permafrost melts (44).Methane can be oxidized in aerobic zones by aerobic methanotrophic bacteria or in anaerobic zones by anaerobic methanotrophic archaea (for a recent review, see reference 27). Anaerobic methane oxidizers were not covered in the context of this study, which focused exclusively on aerobic methanotrophs. These bacteria utilize methane as the sole carbon and energy source through the activity of the enzyme methane monooxygenase (MMO). Most known aerobic methanotrophs are divided into two major groups (type I and type II) based on phylogeny and carbon assimilation pathways (5). Type I methanotrophs, also known as Gammaproteobacteria methanotrophs (6) belong to the family Methylococcaceae within the Gammaproteobacteria subdivision, while type II methanotrophs (Alphaproteobacteria methanotrophs) belong to the family Methylocystaceae in the Alphaproteobacteria subdivision (5). Because of their capacity to oxidize methane, aerobic methanotrophs can significantly reduce methane emissions to the atmosphere and play an important role in the global methane cycle (12, 22). Methanotrophic activity has been observed in cold environments, and methanotrophs might contribute to the reduction of methane emissions from melting permafrost. Aerobic methanotrophic bacteria from cold environments have been reviewed in detail elsewhere (54).Most studies addressing methanotrophs from cold environments were conducted on soils from very few sites located in Northern Europe and Siberia (14, 30, 31, 40, 56-58), while methanotrophic bacterial populations in soils from the Canadian high Arctic remain mostly unexplored (41). In addition, most of these studies were conducted at low latitudes, and the pool of knowledge concerning the activity and diversity of methanotrophic bacterial populations in high Arctic soils is limited. The question being addressed in this study is whether there are active methanotrophs in the active-layer soil in the high Arctic. Therefore, the present work had two objectives: (i) to evaluate the methane oxidation capacity of three active-layer soils from the Canadian high Arctic under various incubation conditions and (ii) to identify and characterize the diversity of the active methanotrophs in these soils using stable isotope probing (SIP) of DNA (DNA-SIP) and sequencing of the 16S rRNA and pmoA genes. With this work, we identify for the first time active methanotrophs in high Arctic soils through the use of DNA-SIP.     

早期鲜卑人和动物骨骼的稳定同位素分析

本文对呼伦贝尔地区扎赉诺尔、团结和东乌珠尔早期鲜卑墓地出土的人和动物骨骼进行了稳定同位素分析,以期揭示早期鲜卑的食物结构和生计方式。C、N稳定同位素分析结果表明,早期鲜卑人的食物来源兼具C_3类和C_4类,其中C_4类食物很可能来自其从事的少量粟作农业。人具有较高的δ~(15)N值,且与动物的δ~(15)N值差异较大,表明人的营养级较高,肉食资源在食物结构中占据较大比重,这可能来自于早期鲜卑人所从事的游牧及狩猎经济的贡献。C、N、S稳定同位素分析结果表明,动物和人的食物来源来自于当地的陆生系统,而个体马(DW M3(1))具有较高的δ~(13)C和δ~(15)N值以及明显不同于其他样品的异常高的δ~(34)S值,说明此个体可能属于来自于其他地质环境的"外来者"。     

Compound-Specific Stable Isotope Analysis: Implications in Hexachlorocyclohexane in-vitro and Field Assessment

Assessment of biotic and abiotic degradation reactions by studying the variation in stable isotopic compositions of organic contaminants in contaminated soil and aquifers is being increasingly considered during the last two decades with development of Compound specific stable isotope analysis (CSIA) technique. CSIA has been recognized as a potential tool for evaluating both qualitative and quantitative degradation with measurement of shifts in isotope ratios of contaminants and their degradation products as its basis. Amongst a wide variety of environmental pollutants including monoaromatics, chlorinated ethenes and benzenes etc., it is only recently that its efficacy is being tested for assessing biodegradation of a noxious pollutant namely hexachlorocyclohexane (HCH), by pure microbial cultures as well as directly at the field site. Anticipating the increase in demand of this technique for monitoring the microbial degradation along with natural attenuation, this review highlights the basic problems associated with HCH contamination emphasizing the applicability of emerging CSIA technique to absolve the major bottlenecks in assessment of HCH. To this end, the review also provides a brief overview of this technique with summarizing the recent revelations put forward by both in vitro and in situ studies by CSIA in monitoring HCH biodegradation.