Scats are often used to study ecological parameters of carnivore species. However, field identification of carnivore scats, based on their morphological characteristics, becomes difficult if many carnivore species are distributed in the same area. We assessed error rates in morphological identification of five sympatric carnivores’ scats in north-eastern Himalayan region of Pakistan during 2013–2017. A sample of 149 scats were subjected to molecular identification using fecal DNA. We used a confusion matrix to assess different types of errors associated with carnivore scat identification. We were able to amplify DNA from 96.6% (n = 144) of scats. Based on field identification of carnivore scats, we had predicted that out of 144 scats: 11 (7.6%) scats were from common leopard, 38 (26.4%) from red fox, 29 (20.1%) from Asiatic jackal, 37 (25.7%) from yellow throated martin, 14 (9.7%) from Asian palm civet and 15 (10.4%) from small Indian civet. However, molecular identification revealed and confirmed nine were scats (6.24%) from common leopard, 40 (27.8 %) from red fox, 21 (14.6%) from Asiatic jackal, 45 (31.25%) from Asian palm civet, 12 (8.3%) scats from small Indian civet, while 11 scats (7.6%) were found from Canis lupus Spp., three (2%) from dog, one (0.7 %) scat sample from porcupine, and two (1.4%) from rhesus monkey. Misidentification rate was highest for Asian palm civet (25.7%), followed by red fox (11.1%) and Asiatic jackal (9.7%) but least for common leopard scats (4.2%). The results specific to our study area concur with previous studies that have recommended that carnivore monitoring programs utilize molecular identification of predator scats. Using only morphological identification of scats can be misleading and may result in wrong management decisions.

The analysis of apex predator diet has the ability to deliver valuable insights into ecosystem health, and the potential impacts a predator might have on commercially relevant species. The Australian sea lion () is an endemic apex predator and one of the world's most endangered pinnipeds. Given that prey availability is vital to the survival of top predators, this study set out to understand what dietary information DNA metabarcoding could yield from 36 sea lion scats collected across 1,500 km of its distribution in southwest Western Australia. A combination of PCR assays were designed to target a variety of potential sea lion prey, including mammals, fish, crustaceans, cephalopods, and birds. Over 1.2 million metabarcodes identified six classes from three phyla, together representing over 80 taxa. The results confirm that the Australian sea lion is a wide-ranging opportunistic predator that consumes an array of mainly demersal fauna. Further, the important commercial species (southern calamari squid) and (western rock lobster) were detected, but were present in <25% of samples. Some of the taxa identified, such as fish, sharks and rays, clarify previous knowledge of sea lion prey, and some, such as eel taxa and two gastropod species, represent new dietary insights. Even with modest sample sizes, a spatial analysis of taxa and operational taxonomic units found within the scat shows significant differences in diet between many of the sample locations and identifies the primary taxa that are driving this variance. This study provides new insights into the diet of this endangered predator and confirms the efficacy of DNA metabarcoding of scat as a noninvasive tool to more broadly define regional biodiversity.

Adaptive radiation can be strongly influenced by interspecific competition for resources, which can lead to diverse outcomes ranging from competitive exclusion to character displacement. In each case, sympatric species are expected to evolve into distinct ecological niches, such as different food types, yet this expectation is not always met when such species are examined in nature. The most common hypotheses to account for the coexistence of species with substantial diet overlap rest on temporal variation in niches (often diets). Yet spatial variation in niche overlap might also be important, pointing to the need for spatiotemporal analyses of diet and diet overlap between closely related species persisting in sympatry. We here perform such an analysis by characterizing the diets of, and diet overlap among, four sympatric Darwin's ground finch species at three sites and over 5 years on a single Galápagos island (Santa Cruz). We find that the different species have broadly similar and overlapping diets - they are to some extent generalists and opportunists - yet we also find that each species retains some 'private' resources for which their morphologies are best suited. Importantly, use of these private resources increased considerably, and diet overlap decreased accordingly, when the availability of preferred shared foods, such as arthropods, was reduced during drought conditions. Spatial variation in food resources was also important. These results together suggest that the ground finches are 'imperfect generalists' that use overlapping resources under benign conditions (in space or time), but then retreat to resources for which they are best adapted during periods of food limitation. These conditions likely promote local and regional coexistence.© 2014 The Authors. Journal of Evolutionary Biology © 2014 European Society For Evolutionary Biology.

Advances in DNA sequencing technology have revolutionized the field of molecular analysis of trophic interactions, and it is now possible to recover counts of food DNA sequences from a wide range of dietary samples. But what do these counts mean? To obtain an accurate estimate of a consumer's diet should we work strictly with data sets summarizing frequency of occurrence of different food taxa, or is it possible to use relative number of sequences? Both approaches are applied to obtain semi-quantitative diet summaries, but occurrence data are often promoted as a more conservative and reliable option due to taxa-specific biases in recovery of sequences. We explore representative dietary metabarcoding data sets and point out that diet summaries based on occurrence data often overestimate the importance of food consumed in small quantities (potentially including low-level contaminants) and are sensitive to the count threshold used to define an occurrence. Our simulations indicate that using relative read abundance (RRA) information often provides a more accurate view of population-level diet even with moderate recovery biases incorporated; however, RRA summaries are sensitive to recovery biases impacting common diet taxa. Both approaches are more accurate when the mean number of food taxa in samples is small. The ideas presented here highlight the need to consider all sources of bias and to justify the methods used to interpret count data in dietary metabarcoding studies. We encourage researchers to continue addressing methodological challenges and acknowledge unanswered questions to help spur future investigations in this rapidly developing area of research.© 2018 John Wiley & Sons Ltd.

Amplified marker-gene sequences can be used to understand microbial community structure, but they suffer from a high level of sequencing and amplification artifacts. The UPARSE pipeline reports operational taxonomic unit (OTU) sequences with ≤1% incorrect bases in artificial microbial community tests, compared with >3% incorrect bases commonly reported by other methods. The improved accuracy results in far fewer OTUs, consistently closer to the expected number of species in a community.

四川山鹧鸪（Arborophila rufipectus）是我国西南山地特有的珍稀濒危鸟类，迄今对其越冬栖息地的了解甚少。2013年1月和2014年1&mdash;2月，在四川老君山国家级自然保护区对其冬季栖息地特征进行了研究。野外共测定了48个利用样方和30个对照样方的特征参数。结果表明：四川山鹧鸪偏好选择海拔较低、坡度平缓、乔木盖度较小、竹子较矮、草本低矮稀疏、1.0 m层盖度较大、4.0~5.0 m层盖度较小、距小路和林缘较近的栖息地。此外，四川山鹧鸪还倾向于选择在距水源较近及地表落叶丰富的栖息地活动。安全与食物条件可能是影响四川山鹧鸪冬季栖息地选择的关键因素。

随着测序技术的快速发展,整合DNA条形码和高通量测序的DNA宏条形码技术已经成为当前研究热点之一,在食草动物的食性鉴定中有很大潜力。放牧动物食性研究是动物营养学和草地生态学领域的重要研究内容。而与传统食性研究方法相比,宏条形码技术可通过对植物DNA条形码的高通量测序,获得样本中的物种组成进而分析动物食性。介绍了传统食性分析手段的局限,重点综述了DNA宏条形码技术的产生、操作原理以及在食草类动物食性鉴定领域中的应用,同时还简述了可能存在的挑战,并对该技术今后的发展方向进行了展望。

Nectogaline shrews are a major component of the small mammalian fauna of Europe and Asia, and are notable for their diverse ecology, including utilization of aquatic habitats. So far, molecular phylogenetic analyses including nectogaline species have been unable to infer a well-resolved, well-supported phylogeny, thus limiting the power of comparative evolutionary and ecological analyses of the group. Here, we employ Bayesian phylogenetic analyses of eight mitochondrial and three nuclear genes to infer the phylogenetic relationships of nectogaline shrews. We subsequently use this phylogeny to assess the genetic diversity within the genus Episoriculus, and determine whether adaptation to aquatic habitats evolved independently multiple times. Moreover, we both analyze the fossil record and employ Bayesian relaxed clock divergence dating analyses of DNA to assess the impact of historical global climate change on the biogeography of Nectogalini. We infer strong support for the polyphyly of the genus Episoriculus. We also find strong evidence that the ability to heavily utilize aquatic habitats evolved independently in both Neomys and Chimarrogale+Nectogale lineages. Our Bayesian molecular divergence analysis suggests that the early history of Nectogalini is characterized by a rapid radiation at the Miocene/Pliocene boundary, thus potentially explaining the lack of resolution at the base of the tree. Finally, we find evidence that nectogalines once inhabited northern latitudes, but the global cooling and desiccating events at the Miocene/Pliocene and Pliocene/Pleistocene boundaries and Pleistocene glaciation resulted in the migration of most Nectogalini lineages to their present day southern distribution.Copyright 2010 Elsevier Inc. All rights reserved.

DNA was extracted from five coprolites, excavated in Gypsum Cave, Nevada and radiocarbon dated to approximately 11 000, 20 000 and 28 500 years BP. All coprolites contained mitochondrial DNA sequences identical to a DNA sequence determined from a bone of the extinct ground sloth Nothrotheriops shastensis. A 157-bp fragment of the chloroplast gene for the large subunit of the ribulosebisphosphate carboxylase (rbcL) was amplified from the boluses and several hundred clones were sequenced. In addition, the same DNA fragment was sequenced from 99 plant species that occur in the vicinity of Gypsum Cave today. When these were compared to the DNA sequences in GenBank, 69 were correctly (two incorrectly) assigned to taxonomic orders. The plant sequences from the five coprolites as well as from one previously studied coprolite were compared to rbcL sequences in GenBank and the contemporary plant species. Thirteen families or orders of plants that formed part of the diet of the Shasta ground sloth could be identified, showing that the ground sloth was feeding on trees as well as herbs and grasses. The plants in the boluses further indicate that the climate 11 000 years BP was dryer than 20 000 and 28 500 years BP. However, the sloths seem to have visited water sources more frequently at 11 000 BP than at earlier times.

Condition indices (CIs) are used in ecological studies as a way of measuring an individual animal’s health and fitness. Noninvasive CIs are estimations of a relative score of fat content or rely on a ratio of body mass compared to some measure of size, usually a linear dimension such as tarsus or snout-vent length. CIs are generally validated invasively by lethal fat extraction as in a seasonal sample of individuals in a population. Many alternatives to lethal fat extraction are costly or time consuming. As an alternative, dual-energy X-ray absorptiometry (DXA) allows for non-destructive analysis of body composition and enables multiple measurements during an animal’s life time. DXA has never been used for ecological studies in a small, free-ranging lizard before, therefore we calibrated this method against a chemical extraction of fat from a sample of 6 geckos (Israeli fan toed gecko Ptyodactylus guttatus) ranging in body mass between 4.2–11.5 g. We then  used this calibrated  DXA measurements to determine the best linear measurement calculated CI for this species.

DNA analysis of predator faeces using high-throughput amplicon sequencing (HTS) enhances our understanding of predator-prey interactions. However, conclusions drawn from this technique are constrained by biases that occur in multiple steps of the HTS workflow. To better characterize insectivorous animal diets, we used DNA from a diverse set of arthropods to assess PCR biases of commonly used and novel primer pairs for the mitochondrial gene, cytochrome oxidase C subunit 1 (COI). We compared diversity recovered from HTS of bat guano samples using a commonly used primer pair "ZBJ" to results using the novel primer pair "ANML." To parameterize our bioinformatics pipeline, we created an arthropod mock community consisting of single-copy (cloned) COI sequences. To examine biases associated with both PCR and HTS, mock community members were combined in equimolar amounts both pre- and post-PCR. We validated our system using guano from bats fed known diets and using composite samples of morphologically identified insects collected in pitfall traps. In PCR tests, the ANML primer pair amplified 58 of 59 arthropod taxa (98%), whereas ZBJ amplified 24-40 of 59 taxa (41%-68%). Furthermore, in an HTS comparison of field-collected samples, the ANML primers detected nearly fourfold more arthropod taxa than the ZBJ primers. The additional arthropods detected include medically and economically relevant insect groups such as mosquitoes. Results revealed biases at both the PCR and sequencing levels, demonstrating the pitfalls associated with using HTS read numbers as proxies for abundance. The use of an arthropod mock community allowed for improved bioinformatics pipeline parameterization.© 2018 John Wiley & Sons Ltd.

Bats are well known for species richness and ecological diversity, and thus, they provide a good opportunity to study relationships and interaction between species. To assess interactions, we consider distinct traits that are probably to be triggered by niche shape and evolutionary processes. We present data on the trophic niche differentiation between two sympatric European trawling bat species, Myotis dasycneme and Myotis daubentonii, incorporating a wide spectrum of methodological approaches. We measure morphological traits involved in foraging and prey handling performance including bite force, weightlifting capacity and wing morphology. We then measure resulting prey consumption using both morphological and molecular diet analyses. These species closely resemble each other in morphological traits, however, subtle but significant differences were apparent in bite force and lift capacity, which are related to differences in basic body and head size. Both morphological and molecular diet analyses show strong niche overlap. We detected subtle differences in less frequent prey items, as well as differences in the exploitation of terrestrial and aquatic-based prey groups. Myotis dasycneme feeds more on aquatic prey, like Chironomidae and their pupal stages, or on the aquatic moth Acentria ephemerella. Myotis daubentonii feeds more on terrestrial prey, like Brachycera, or Coleoptera. This suggests that these bats use different microhabitats within the habitat where they co-occur.© 2013 John Wiley & Sons Ltd.

As the apex predator of plateau ecosystems in Central Asia and the Qinghai-Tibet Plateau, the snow leopard (Panthera uncia) plays an essential role in maintaining food-web structure and ecosystem stability. Learning the diet composition and dynamics of the snow leopard is important for understanding its role in ecosystem functioning and interspecific interactions. Previous diet analyses of the snow leopard have been based mainly on morphological identification of food debris in the feces, though the accuracy of this practice has been broadly debated. The Qionglai Mountains are located at the southeast edge of the snow leopard range, harboring a small and relatively isolated population of snow leopards that are barely studied. Using non-invasive sampling, we collected 38 putative snow leopard fecal samples in the Wolong National Nature Reserve in the Qionglai Mountains. To identify the fecal origin, we extracted the fecal DNA and amplified the mitochondrial DNA 16S rRNA gene fragment. Twenty-two fecal samples were identified as originating from snow leopards. Subsequently, vertebrate universal primers and a snow leopard-specific blocking oligo were used to amplify the food components in the fecal DNA, and then high-throughput sequencing was performed to analyze the diet composition of snow leopards. The blue sheep (Pseudois nayaur) was detected in 67% of the samples and was found to be the main staple food of snow leopards’ diet. The domestic yak (Bos grunniens) appeared in 33% of the fecal samples, also accounting for a high proportion of the snow leopard diet. In addition, pikas (Ochotona spp.) and birds were found in a small number of fecal samples. Therefore, wild prey was found to be the main food source for snow leopards in Wolong. However, livestock (yak) also accounted for a relatively large proportion of their diet.

作为中亚和青藏高原山地生态系统中的顶级捕食者, 雪豹(Panthera uncia)对于维持食物网结构和生态系统稳定性有重要作用。了解雪豹的食性组成和变化对于理解其生态系统功能和物种间相互作用有重要意义。以往的雪豹食性分析多基于对其粪便中食物残渣的形态学鉴定, 但准确度受人员经验和主观因素影响较大。邛崃山脉位于雪豹分布区东南缘, 该区域的雪豹种群规模小且相对孤立, 研究匮乏。本研究基于非损伤性取样, 在邛崃山脉的卧龙国家级保护区采集疑似雪豹粪便样品38份, 首先提取粪便DNA, 并扩增线粒体DNA 16S rRNA基因片段进行分子物种鉴定, 确定其中22份为雪豹粪便样品。随后, 利用脊椎动物通用引物和雪豹特异性阻抑引物扩增粪便DNA中的食物成分, 并进行高通量测序, 分析雪豹食性构成。食性分析结果显示岩羊(Pseudois nayaur)是卧龙地区雪豹最主要的食物, 在67%的样品中均有检出。家牦牛(Bos grunniens)在33%的样品中出现, 也在雪豹食性中占较高比例。此外, 鼠兔(Ochotona spp.)和鸟类也在少量样品中发现。可见, 野生猎物是卧龙地区雪豹的主要食物资源; 与大世界大多数其他地区的雪豹食性相同, 野生大型有蹄类是雪豹最重要的食物。然而家畜(牦牛)在卧龙雪豹食谱中有相当高的占比, 显示该区域内可能存在较为严重的由雪豹捕食散养家畜引起的人兽冲突问题。

The analysis of food webs and their dynamics facilitates understanding of the mechanistic processes behind community ecology and ecosystem functions. Having accurate techniques for determining dietary ranges and components is critical for this endeavour. While visual analyses and early molecular approaches are highly labour intensive and often lack resolution, recent DNA-based approaches potentially provide more accurate methods for dietary studies. A suite of approaches have been used based on the identification of consumed species by characterization of DNA present in gut or faecal samples. In one approach, a standardized DNA region (DNA barcode) is PCR amplified, amplicons are sequenced and then compared to a reference database for identification. Initially, this involved sequencing clones from PCR products, and studies were limited in scale because of the costs and effort required. The recent development of next generation sequencing (NGS) has made this approach much more powerful, by allowing the direct characterization of dozens of samples with several thousand sequences per PCR product, and has the potential to reveal many consumed species simultaneously (DNA metabarcoding). Continual improvement of NGS technologies, on-going decreases in costs and current massive expansion of reference databases make this approach promising. Here we review the power and pitfalls of NGS diet methods. We present the critical factors to take into account when choosing or designing a suitable barcode. Then, we consider both technical and analytical aspects of NGS diet studies. Finally, we discuss the validation of data accuracy including the viability of producing quantitative data.© 2011 Blackwell Publishing Ltd.

Sympatric cryptic species, characterized by low morphological differentiation, pose a challenge to understanding the role of interspecific competition in structuring ecological communities. We used traditional (morphological) and novel molecular methods of diet analysis to study the diet of two cryptic bat species that are sympatric in southern England (Plecotus austriacus and P. auritus) (Fig. 1). Using Roche FLX 454 (Roche, Basel, CH) high-throughput sequencing (HTS) and uniquely tagged generic arthropod primers, we identified 142 prey Molecular Operational Taxonomic Units (MOTUs) in the diet of the cryptic bats, 60% of which were assigned to a likely species or genus. The findings from the molecular study supported the results of microscopic analyses in showing that the diets of both species were dominated by lepidopterans. However, HTS provided a sufficiently high resolution of prey identification to determine fine-scale differences in resource use. Although both bat species appeared to have a generalist diet, eared-moths from the family Noctuidae were the main prey consumed. Interspecific niche overlap was greater than expected by chance (O(jk) = 0.72, P < 0.001) due to overlap in the consumption of the more common prey species. Yet, habitat associations of nongeneralist prey species found in the diets corresponded to those of their respective bat predator (grasslands for P. austriacus, and woodland for P. auritus). Overlap in common dietary resource use combined with differential specialist prey habitat associations suggests that habitat partitioning is the primary mechanism of coexistence. The performance of HTS is discussed in relation to previous methods of molecular and morphological diet analysis. By enabling species-level identification of dietary components, the application of DNA sequencing to diet analysis allows a more comprehensive comparison of the diet of sympatric cryptic species, and therefore can be an important tool for determining fine-scale mechanisms of coexistence.

VSEARCH is an open source and free of charge multithreaded 64-bit tool for processing and preparing metagenomics, genomics and population genomics nucleotide sequence data. It is designed as an alternative to the widely used USEARCH tool (Edgar, 2010) for which the source code is not publicly available, algorithm details are only rudimentarily described, and only a memory-confined 32-bit version is freely available for academic use.

Defining historical baselines is critical for species conservation. Under the niche reduction hypothesis, species in decline may be restricted disproportionately from parts of their environmental niche. This bias likely has important implications for modeling species' distributions if only contemporary occurrences (i.e. post-range reduction) are used, because suitable habitat will be classified as unsuitable. Unfortunately, robust historical occurrence data is rarely available for sensitive species. In this study, we documented historical locations of the endangered, keystone giant kangaroo rat Dipodomys ingens by examining aerial imagery for burrow mounds. These burrow mounds are readily identifiable and distinguishable from other soil disturbances. We found giant kangaroo rat burrows well outside the currently accepted estimate of their historical distribution. Following the niche reduction hypothesis, we found that giant kangaroo rats have been extirpated from the flattest, hottest, driest parts of their range due to agricultural conversion. This reduction in their realized niche led to significant changes between historical and contemporary models of their distribution. We found that giant kangaroo rats may have occupied up to 56% more habitat historically than currently believed. Our results provide new guidance for managers working on restoration and habitat protection for this ecosystem engineer. This study highlights the critical importance of modeling historical distributions using the entire environmental niche once occupied by species of conservation need.

Large carnivores play an important role in the regulation of food-web structure and ecosystem functioning. However, large carnivores face serious threats that have caused declines in their populations and geographic ranges due to habitat loss and degradation, hunting, human disturbance and pathogen transmission. Conservation of large carnivore species richness and population size has become a pressing issue and an important research focus of conservation biology. The western Sichuan Plateau, located at the intersection of the mountains of southwest China and the eastern margin of the Tibetan Plateau, is a global biodiversity hotspot and has high carnivore species richness. However, increasing human activities may exacerbate the destruction of local flora and fauna, thereby threatening the survival of wild carnivores. Information on species composition and dietary habits can improve our understanding of the structure and function of the ecosystem and food-web relationships in the study area. In addition, species composition and dietary habits are of great significance for understanding multi-species coexistence mechanisms and preserving biodiversity. This study collected carnivore fecal samples from Xinlong and Shiqu counties in the Ganzi Tibetan Autonomous Prefecture, Sichuan Province. DNA was then extracted from the samples and the species was identified based on DNA sequences and DNA barcoding techniques. Seven carnivores were identified, including five large carnivores (Canis lupus, Ursus arctos, Panthera pardus, P. uncia and Canis lupus familiaris) and two medium and small-sized carnivores (Prionailurus bengalensis and Vulpes vulpes). Using fecal DNA, high-throughput sequencing and metabarcoding, we conducted diet analysis for the seven carnivores and found 28 different food molecular operational taxonomic units (MOTUs), including 19 mammals, eight birds and one fish species. The predominant prey categories of wolves, dogs and brown bears were ungulates. The domestic yak (Bos grunniens) was the most frequently identified prey species. Small mammals such as rodents and lagomorphs accounted for a significant proportion in the diets of leopard cats and red foxes, The most frequent prey of this category of carnivore were the Chinese scrub vole (Neodon irene) and plateau pika (Ochotona curzoniae). In addition, leopards and snow leopards mainly fed on the Chinese goral (Naemorhedus griseus) and blue sheep (Pseudois nayaur), respectively. Our study highlights the utility of fecal DNA and metabarcoding technique in fast carnivore surveys and high-throughput diet analysis, and provides a technical reference and guidance for future biodiversity surveys and food-web studies.

在陆地生态系统中, 大型食肉动物对于稳定食物网结构和生态系统功能有重要作用。在世界范围内, 由于栖息地丧失和破碎化、猎杀、人类活动干扰以及病原体的传播, 大型食肉动物生存正面临严重威胁, 多种食肉动物地理分布范围及种群数量大幅度缩减。如何有效保护大型食肉动物物种多样性及种群已经成为世界关注的焦点问题和保护生物学的重要研究方向。川西高原地处我国西南山地与青藏高原东缘交界地带, 属于世界生物多样性热点地区, 是世界大型食肉动物物种最丰富的地区之一, 而日益增强的人类活动可能会加剧对当地动植物资源的破坏, 进而威胁野生食肉动物的生存。获得准确的物种多样性信息及食肉动物食性数据有助于深入了解该地区生态系统结构及食物网关系, 对研究物种共存机制及生物多样性保护有重要意义。本研究通过从四川甘孜藏族自治州新龙县和石渠县野外采集的食肉动物粪便样品中提取DNA, 利用DNA条形码进行物种鉴定, 快速获得该地区食肉动物物种构成信息。38份粪便样品经鉴定来自于7种食肉动物, 分别为5种大型食肉动物(狼Canis lupus、棕熊Ursus arctos、豹Panthera pardus、雪豹P. unica、狗Canis lupus familiaris)和2种中小型食肉动物(豹猫Prionailurus bengalensis、赤狐Vulpes vulpes)。进一步利用高通量测序和宏条形码技术对7种食肉动物粪便中的食物DNA进行精准食性分析, 得到包含19种哺乳类、8种鸟类和1种鱼类共计28个不同的食物分子可操作分类单元(molecular operational taxonomic unit, MOTU)。结果显示, 狼、狗、棕熊最主要的食物来源为偶蹄目动物, 其中取食频率最高的物种为家牦牛(Bos grunniens); 而豹猫和赤狐食物中小型哺乳动物如啮齿目和兔形目占重要比例, 其中高原松田鼠(Neodon irene)和高原鼠兔(Ochotona curzoniae)被取食频率最高。豹和雪豹的食物分别为偶蹄目的中华斑羚(Naemorhedus griseus)和岩羊(Pseudois nayaur)。本研究显示了粪便DNA及宏条形码技术在食肉动物多样性快速调查及高通量精确食性分析中的应用前景, 并为此类研究提供了技术路线的有力借鉴。

Rather little is known about the dietary richness and variation of generalist insectivorous species, including birds, due primarily to difficulties in prey identification. Using faecal metabarcoding, we provide the most comprehensive analysis of a passerine's diet to date, identifying the relative magnitudes of biogeographic, habitat and temporal trends in the richness and turnover in diet of Cyanistes caeruleus (blue tit) along a 39 site and 2° latitudinal transect in Scotland. Faecal samples were collected in 2014-2015 from adult birds roosting in nestboxes prior to nest building. DNA was extracted from 793 samples and we amplified COI and 16S minibarcodes. We identified 432 molecular operational taxonomic units that correspond to putative dietary items. Most dietary items were rare, with Lepidoptera being the most abundant and taxon-rich prey order. Here, we present a statistical approach for estimation of gradients and intersample variation in taxonomic richness and turnover using a generalised linear mixed model. We discuss the merits of this approach over existing tools and present methods for model-based estimation of repeatability, taxon richness and Jaccard indices. We found that dietary richness increases significantly as spring advances, but changes little with elevation, latitude or local tree composition. In comparison, dietary composition exhibits significant turnover along temporal and spatial gradients and among sites. Our study shows the promise of faecal metabarcoding for inferring the macroecology of food webs, but we also highlight the challenge posed by contamination and make recommendations of laboratory and statistical practices to minimise its impact on inference.© 2020 John Wiley & Sons Ltd.

Virtually all empirical ecological studies require species identification during data collection. DNA metabarcoding refers to the automated identification of multiple species from a single bulk sample containing entire organisms or from a single environmental sample containing degraded DNA (soil, water, faeces, etc.). It can be implemented for both modern and ancient environmental samples. The availability of next-generation sequencing platforms and the ecologists' need for high-throughput taxon identification have facilitated the emergence of DNA metabarcoding. The potential power of DNA metabarcoding as it is implemented today is limited mainly by its dependency on PCR and by the considerable investment needed to build comprehensive taxonomic reference libraries. Further developments associated with the impressive progress in DNA sequencing will eliminate the currently required DNA amplification step, and comprehensive taxonomic reference libraries composed of whole organellar genomes and repetitive ribosomal nuclear DNA can be built based on the well-curated DNA extract collections maintained by standardized barcoding initiatives. The near-term future of DNA metabarcoding has an enormous potential to boost data acquisition in biodiversity research.© 2012 Blackwell Publishing Ltd.

Food availability and diet selection are important factors influencing the abundance and distribution of wild waterbirds. In order to better understand changes in waterbird population, it is essential to figure out what they feed on. However, analyzing their diet could be difficult and inefficient using traditional methods such as microhistologic observation. Here, we addressed this gap of knowledge by investigating the diet of greater white-fronted gooseAnser albifronsand bean gooseAnser fabalis, which are obligate herbivores wintering in China, mostly in the Middle and Lower Yangtze River floodplain. First, we selected a suitable and high-resolution marker gene for wetland plants that these geese would consume during the wintering period. Eight candidate genes were included:rbcL,rpoC1,rpoB,matK,trnH-psbA,trnL (UAA),atpF-atpH, andpsbK-psbI. The selection was performed via analysis of representative sequences from NCBI and comparison of amplification efficiency and resolution power of plant samples collected from the wintering area. ThetrnL gene was chosen at last with c/h primers, and a local plant reference library was constructed with this gene. Then, utilizing DNA metabarcoding, we discovered 15 food items in total from the feces of these birds. Of the 15 unique dietary sequences, 10 could be identified at specie level. As for greater white-fronted goose, 73% of sequences belonged toPoaceaespp., and 26% belonged toCarexspp. In contrast, almost all sequences of bean goose belonged toCarexspp. (99%). Using the same samples, microhistology provided consistent food composition with metabarcoding results for greater white-fronted goose, while 13% ofPoaceaewas recovered for bean goose. In addition, two other taxa were discovered only through microhistologic analysis. Although most of the identified taxa matched relatively well between the two methods, DNA metabarcoding gave taxonomically more detailed information. Discrepancies were likely due to biased PCR amplification in metabarcoding, low discriminating power of current marker genes for monocots, and biases in microhistologic analysis. The diet differences between two geese species might indicate deeper ecological significance beyond the scope of this study. We concluded that DNA metabarcoding provides new perspectives for studies of herbivorous waterbird diets and inter-specific interactions, as well as new possibilities to investigate interactions between herbivores and plants. In addition, microhistologic analysis should be used together with metabarcoding methods to integrate this information.

研究啮齿动物的食性及其营养生态位特征不仅可以了解高寒草甸环境变化下物种的食性适应特征，还可探讨多种啮齿动物的种间关系。本研究采用粪便显微组织观察法，分析了甘南草原高原鼠兔（Ochotona curzoniae）、高原鼢鼠（Eospalax baileyi）和喜马拉雅旱獭（Marmota himalayana）3种啮齿动物的食性及营养生态位特征。结果发现：3种啮齿动物所采食的植物种类基本相同，但其比例各异；高原鼢鼠所采食的植物主要是珠芽蓼（Polygonum viviparum）和鹅绒委陵菜（Potentilla anserina）等杂类草，且对莎草科（Cyperaceae）植物的采食量显著高于其它两种动物，高原鼠兔主要采食禾本科（Poaceae）植物，喜马拉雅旱獭则主要采食禾本科和菊科（Asteraceae）植物；营养生态位宽度依次为喜马拉雅旱獭>高原鼢鼠>高原鼠兔，其中高原鼠兔与喜马拉雅旱獭的营养生态位重叠最大（0.52）。3种啮齿动物在食物上存在分化，营养生态位分离，是其共存的重要原因。