Lipid extraction has little effect on the delta N-15 of aquatic consumers

Proper application of stable isotopes (e. g., delta N-15 and delta C-13) to food web analysis requires an understanding of all nondietary factors that contribute to isotopic variability. Lipid extraction is often used during stable isotope analysis (SIA), because synthesized lipids have a low delta C-13 and can mask the delta C-13 of a consumer's diet. Recent studies indicate that lipid extraction intended to adjust delta C-13 may also cause shifts in delta N-15, but the magnitude of and reasons for the shift are highly uncertain. We examined a large data set (n = 854) for effects of lipid extraction (using Bligh and dyer's [ 1959] chloroform-methanol solvent mixtures) on the delta N-15 of aquatic consumers. We found no effect of chemically extracting lipids on the delta N-15 of whole zooplankton, unionid mussels, and fish liver samples, and found a small increase in fish muscle delta N-15 of similar to 0.4%. We also detected a negative relationship between the shift in delta N-15 following extraction and the C:N ratio in muscle tissue, suggesting that effects of extraction were greater for tissue with lower lipid content. As long as appropriate techniques such as those from Bligh and dyer (1959) are used, effects of lipid extraction on delta N-15 of aquatic consumers need not be a major consideration in the SIA of food webs.

Phenotype-environment correlations in a putative whitefish adaptive radiation

1. The adaptive radiation of fishes into benthic (littoral) and pelagic (lentic) morphs in post-glaciallakes has become an important model system for speciation. Although these systems are well stud-ied, there is little evidence of the existence of morphs that have diverged to utilize resources in theremaining principal lake habitat, the profundal zone.
2. Here, we tested phenotype-environment correlations of three whitefish (Coregonus lavaretus)morphs that have radiated into littoral, pelagic and profundal niches in northern Scandinavianlakes. We hypothesized that morphs in such trimorphic systems would have a morphology adaptedto one of the principal lake habitats (littoral, pelagic or profundal niches). Most whitefish popula-tions in the study area are formed by a single (monomorphic) whitefish morph, and we furtherhypothesized that these populations should display intermediate morphotypes and niche utiliza-tion. We used a combination of traditional (stomach content, habitat use, gill raker counts) andmore recently developed (stable isotopes, geometric morphometrics) techniques to evaluate pheno-type-environment correlations in two lakes with trimorphic and two lakes with monomorphicwhitefish.
3. Distinct phenotype-environment correlations were evident for each principal niche in whitefishmorphs inhabiting trimorphic lakes. Monomorphic whitefish exploited multiple habitats, hadintermediate morphology, displayed increased variance in gillraker-counts, and relied significantlyon zooplankton, most likely due to relaxed resource competition.
4. We suggest that the ecological processes acting in the trimorphic lakes are similar to each other,and are driving the adaptive evolution of whitefish morphs, possibly leading to the formation ofnew species.

Rapid sympatric ecological differentiation of crater lake cichlid fishes within historic times

Background: After a volcano erupts, a lake may form in the cooled crater and become an isolated aquatic ecosystem. This makes fishes in crater lakes informative for understanding sympatric evolution and ecological diversification in barren environments. From a geological and limnological perspective, such research offers insight about the process of crater lake ecosystem establishment and speciation. In the present study we use genetic and coalescence approaches to infer the colonization history of Midas cichlid fishes (Amphilophus cf. citrinellus) that inhabit a very young crater lake in Nicaragua-the ca. 1800 year-old Lake Apoyeque. This lake holds two sympatric, endemic morphs of Midas cichlid: one with large, hypertrophied lips (~20% of the total population) and another with thin lips. Here we test the associated ecological, morphological and genetic diversification of these two morphs and their potential to represent incipient speciation.
Results: Gene coalescence analyses [11 microsatellite loci and mitochondrial DNA (mtDNA) sequences] suggest that crater lake Apoyeque was colonized in a single event from the large neighbouring great lake Managua only about 100 years ago. This founding in historic times is also reflected in the extremely low nuclear and mitochondrial genetic diversity in Apoyeque. We found that sympatric adult thin- and thick-lipped fishes occupy distinct ecological trophic niches. Diet, body shape, head width, pharyngeal jaw size and shape and stable isotope values all differ significantly between the two lip-morphs. The eco-morphological features pharyngeal jaw shape, body shape, stomach contents and stable isotopes (d15N) all show a bimodal distribution of traits, which is compatible with the expectations of an initial stage of ecological speciation under disruptive selection. Genetic differentiation between the thin- and thick-lipped population is weak at mtDNA sequence (FST = 0.018) and absent at nuclear microsatellite loci (FST < 0.001).
Conclusions: This study provides empirical evidence of eco-morphological differentiation occurring very quickly after the colonization of a new and vacant habitat. Exceptionally low levels of neutral genetic diversity and inference from coalescence indicates that the Midas cichlid population in Apoyeque is much younger (ca. 100 years or generations old) than the crater itself (ca. 1 800 years old). This suggests either that the crater remained empty for many hundreds of years after its formation or that remnant volcanic activity prevented the establishment of a stable fish population during the early life of the crater lake. Based on our findings of eco-morphological variation in the Apoyeque Midas cichlids, and known patterns of adaptation in Midas cichlids in general, we suggest that this population may be in a very early stage of speciation (incipient species), promoted by disruptive selection and ecological diversification.

delta C-13 and delta N-15 reveal significant differences in the coastal foodwebs of the seas surrounding Trinidad and Tobago

This study assessed nearshore, marine ecosystem function around Trinidad and Tobago (TT). The coastline of TT is highly complex, bordered by the Atlantic Ocean, the Caribbean Sea, the Gulf of Paria and the Columbus Channel, and subject to local terrestrial runoff and regional riverine inputs (e.g. the Orinoco and Amazon rivers). Coastal organisms can assimilate energy from allochthonous and autochthonous Sources, We assessed whether stable isotopes delta C-13 and delta N-15 Could be used to provide a rapid assessment of trophic interactions in primary consumers around the islands. Filter-feeding (bivalves and barnacles) and grazing organisms (gastropods and chitons) were collected from 40 marine sites during the wet season. The flesh of organisms was analysed for delta C-13 and delta N-15. Results indicate significant variation in primary consumers (by feeding guild and sampling zone). This variation was linked to different energy Sources being assimilated by consumers. Results suggest that offshore production is fuelling intertidal foodwebs; for example, a depleted delta C-13 signature in grazers from the Gulf of Paria, Columbus Channel and the Caribbean and Atlantic coastline of 9 Tobago indicates that carbon with an offshore origin (e.g. phytoplankton and dissolved organic matter) is more important than benthic or littoral algae (luring the wet season. Results also confirm findings from other studies indicating that much of the coastline is subject to Cultural eutrophication. This Study revealed that ecosystem function is spatially variable around the coastline of TT, This has clear implications for marine resource management, as a single management approach is unlikely to be successful at a national level.

Rock hyrax middens: A palaeoenvironmental archive for southern African drylands

Like many of the world's subtropical regions, southern Africa is highly sensitive to changes in the earth's climate system, but a dearth of reliable palaeoenvironmental records means that relatively little is known about how regional environments have been affected over centennial to multi-millennial timescales. To a large extent this sensitivity is a function of the position of these regions at the interface between temperate and tropical circulation systems. The resulting seasonality and irregularity of rainfall have limited the development of suitable archives, such as lakes and wetlands, for the preservation of palaeoenvironmental proxies.

This paper reviews and evaluates the value of rock hyrax middens as novel palaeoenvironmental archives in southern Africa. Considered are (1) the contemporary taxonomy, distribution and ecology of hyraxes, (2) the mechanisms of hyrax midden development, their physical and chemical structure, rates of accumulation and age; and (3) the palaeoenvironmental proxies preserved within hyrax middens, including fossil pollen, stable isotopes and biomarkers. The interpretive constraints and opportunities offered by these various midden characteristics are assessed with a view to demonstrating the potential of these deposits, widespread as they are through arid and semi-arid southern Africa, in providing a more detailed and chronologically resolved view of late Quaternary palaeoenvironments across the subcontinent. (C) 2012 Elsevier Ltd. All rights reserved.

Parallel and nonparallel ecological, morphological and genetic divergence in lake-stream stickleback from a single catchment

Parallel phenotypic evolution in similar environments has been well studied in evolutionary biology; however, comparatively little is known about the influence of determinism and historical contingency on the nature, extent and generality of this divergence. Taking advantage of a novel system containing multiple lake-stream stickleback populations, we examined the extent of ecological, morphological and genetic divergence between three-spined stickleback present in parapatric environments. Consistent with other lake-stream studies, we found a shift towards a deeper body and shorter gill rakers in stream fish. Morphological shifts were concurrent with changes in diet, indicated by both stable isotope and stomach contents analysis. Performing a multivariate test for shared and unique components of evolutionary response to the distance gradient from the lake, we found a strong signature of parallel adaptation. Nonparallel divergence was also present, attributable mainly to differences between river locations. We additionally found evidence of genetic substructuring across five lake-stream transitions, indicating that some level of reproductive isolation occurs between populations in these habitats. Strong correlations between pairwise measures of morphological, ecological and genetic distance between lake and stream populations supports the hypothesis that divergent natural selection between habitats drives adaptive divergence and reproductive isolation. Lake-stream stickleback divergence in Lough Neagh provides evidence for the deterministic role of selection and supports the hypothesis that parallel selection in similar environments may initiate parallel speciation.

Parsing parallel evolution:Ecological divergence and differential gene expression in the adaptive radiations of thick-lipped Midas cichlid fishes from Nicaragua

The study of parallel evolution facilitates the discovery of common rules of diversification. Here, we examine the repeated evolution of thick lips in Midas cichlid fishes (the Amphilophus citrinellus species complex) - from two Great Lakes and two crater lakes in Nicaragua - to assess whether similar changes in ecology, phenotypic trophic traits and gene expression accompany parallel trait evolution. Using next-generation sequencing technology, we characterize transcriptome-wide differential gene expression in the lips of wild-caught sympatric thick- and thin-lipped cichlids from all four instances of repeated thick-lip evolution. Six genes (apolipoprotein D, myelin-associated glycoprotein precursor, four-and-a-half LIM domain protein 2, calpain-9, GTPase IMAP family member 8-like and one hypothetical protein) are significantly underexpressed in the thick-lipped morph across all four lakes. However, other aspects of lips' gene expression in sympatric morphs differ in a lake-specific pattern, including the magnitude of differentially expressed genes (97-510). Generally, fewer genes are differentially expressed among morphs in the younger crater lakes than in those from the older Great Lakes. Body shape, lower pharyngeal jaw size and shape, and stable isotopes (dC and dN) differ between all sympatric morphs, with the greatest differentiation in the Great Lake Nicaragua. Some ecological traits evolve in parallel (those related to foraging ecology; e.g. lip size, body and head shape) but others, somewhat surprisingly, do not (those related to diet and food processing; e.g. jaw size and shape, stable isotopes). Taken together, this case of parallelism among thick- and thin-lipped cichlids shows a mosaic pattern of parallel and nonparallel evolution. © 2012 Blackwell Publishing Ltd.

Dietary switching of collembola in grassland soil food webs

Soil food webs are characterised by complex direct and indirect effects among the organisms. Consumption of microorganisms by soil animals is considered as an important factor that contributes to the stability of communities, though cascading effects within the food web can be difficult to detect. In a greenhouse experiment, an addition of a high number the fungal feeding collembola Folsomia quadrioculata was applied to grassland soil food webs in monocultures of three plant species: Plantago lanceolato (forb), Lotus corniculatus (legume) and Holcus lanatus (grass). The abundance of microorganisms, determined as the abundances of phospholipid fatty acids (PLFAs) and the abundances of resident invertebrates, nematodes and collembolans, did not change due to the addition of E quadrioculata. Trophic positions of collembolans were determined by analyses of natural abundances of N-15 stable isotopes. The use of food resources by microorganisms and collembolans was determined by C-13 analysis of microbial PLFAs and solid samples of collembolans. delta C-13 values of the resident collembola Folsomia fimetaria were lower in the presence of E quadrioculata than in the control food webs indicating a use of more depleted C-13 food resources by E fimetaria. The delta N-15 values of E fimetaria did not change at the addition of E quadrioculata thus no change of trophic levels was detected. The switch of E fimetaria to a different food resource could be due to indirect interactions in the food web as the two collembolan species were positioned on different trophic positions, according to different delta N-15 values. (c) 2008 Elsevier Ltd. All rights reserved.

A composite pollen-based stratotype for inter-regional evaluation of climatic events in New Zealand over the past 30,000 years (NZ-INTIMATE project)

Our review of paleoclimate information for New Zealand pertaining to the past 30,000 years has identified a general sequence of climatic events, spanning the onset of cold conditions marking the final phase of the Last Glaciation, through to the emergence to full interglacial conditions in the early Holocene. In order to facilitate more detailed assessments of climate variability and any leads or lags in the timing of climate changes across the region, a composite stratotype is proposed for New Zealand. The stratotype is based on terrestrial stratigraphic records and is intended to provide a standard reference for the intercomparison and evaluation of climate proxy records. We nominate a specific stratigraphic type record for each climatic event, using either natural exposure or drill core stratigraphic sections. Type records were selected on thebasis of having very good numerical age control and a clear proxy record. In all cases the main proxy of the type record is subfossil pollen. The type record for the period from ca 30 to ca 18 calendar kiloyears BP (cal. ka BP) is designated in lake-bed sediments from a small morainic kettle lake (Galway tarn) in western South Island. The Galway tarn type record spans a period of full glacial conditions (Last Glacial Coldest Period, LGCP) within the Otira Glaciation, and includes three cold stadials separated by two cool interstadials. The type record for the emergence from glacial conditions following the termination of the Last Glaciation (post-Termination amelioration) is in a core of lake sediments from a maar (Pukaki volcanic crater) in Auckland, northern North Island, and spans from ca 18 to 15.64±0.41 cal. ka BP. The type record for the Lateglacial period is an exposure of interbedded peat and mud at montane Kaipo bog, eastern North Island. In this high-resolution type record, an initial mild period was succeeded at 13.74±0.13 cal. ka BP by a cooler period, which after 12.55±0.14 cal. ka BP gave way to a progressive ascent to full interglacial conditions that were achieved by 11.88±0.18 cal. ka BP. Although a type section is not formally designated for the Holocene Interglacial (11.88±0.18 cal. ka BP to the present day), the sedimentary record of Lake Maratoto on the Waikato lowlands, northwestern North Island, is identified as a prospective type section pending the integration and updating of existing stratigraphic and proxy datasets, and age models. The type records are interconnected by one or more dated tephra layers, the ages of which are derived from Bayesian depositional modelling and OxCal-based calibrations using the IntCal09 dataset. Along with the type sections and the Lake Maratoto record, important, well-dated terrestrial reference records are provided for each climate event. Climate proxies from these reference records include pollen flora, stable isotopes from speleothems, beetle and chironomid fauna, and glacier moraines. The regional composite stratotype provides a benchmark against which to compare other records and proxies. Based on the composite stratotype, we provide an updated climate event stratigraphic classification for the New Zealand region. © 2013 Elsevier Ltd.

Determination of geographical origin of distillers dried grains and solubles using isotope ratio mass spectrometry

In recent years distillers dried grains and solubles (DDGS), co-products of the bio-ethanol and beverages industries, have become globally traded commodity for the animal feed sector. As such it is becoming increasingly important to be able to trace the geographical origin of commodities in case of a contamination incident or authenticity issue arise. In this study, 137 DDGS samples from a range of different geographical origins (China, USA, Canada and European Union) were collected and analyzed. Isotope ratio mass spectrometry (IRMS) was used to analyze the DDGS for ²H/¹H, ¹³C/¹²C, ¹⁵N/¹⁴N, ¹⁸O/¹⁶O and ³⁴S/³²S isotope ratios which can vary depending on geographical origin and processing. Univariate and multivariate statistical techniques were employed to investigate the feasibility of using the IRMS data to determine botanical and geographical origin of the DDGS. The results indicated that this commodity could be differentiated according to their place of origin by the analysis of stable isotopes of hydrogen, carbon, nitrogen and oxygen but not with sulfur. By adding data to the models produced in this study, potentially an isotope databank could be set up for traceability procedures for DDGS, similar to the one established already for wine which will help in feed and food security issues arising worldwide.

Evaluation of methodologies to determine vegetable oil species present in oil mixtures: Proposition of an approach to meet the EU legislation demands for correct vegetable oils labelling

Refined vegetable oils are widely used in the food industry as ingredients or components in many processed food products in the form of oil blends. To date, the generic term 'vegetable oil' has been used in the labelling of food containing oil blends. With the introduction of new EU Regulation for Food Information (1169/2011) due to take effect in 2014, the oil species used must be clearly identified on the package and there is a need for development of fit for purpose methodology for industry and regulators alike to verify the oil species present in a product. The available methodologies that may be employed to authenticate the botanical origin of a vegetable oil admixture were reviewed and evaluated. The majority of the sources however, described techniques applied to crude vegetable oils such as olive oil due to the lack of refined vegetable oil focused studies. Nevertheless, DNA based typing methods and stable isotopes procedures were found not suitable for this particular purpose due to several issues. Only a small number of specific chromatographic and spectroscopic fingerprinting methods in either targeted or untargeted mode were found to be applicable in potentially providing a solution to this complex authenticity problem. Applied as a single method in isolation, these techniques would be able to give limited information on the oils identity as signals obtained for various oil types may well be overlapping. Therefore, more complex and combined approaches are likely to be needed to identify the oil species present in oil blends employing a stepwise approach in combination with advanced chemometrics. Options to provide such a methodology are outlined in the current study.

Air pollutant contamination and acidification of surface waters in the North York Moors, UK: Multi-proxy evidence from the sediments of a moorland pool

Despite the extensive geographical range of palaeolimnological studies designed to assess the extent of surface water acidification in the United Kingdom during the 1980s, little attention was paid to the status of surface waters in the North York Moors (NYM). In this paper, we present sediment core data from a moorland pool in the NYM that provide a record of air pollution contamination and surface water acidification. The 41-cm-long core was divided into three lithostratigraphic units. The lower two comprise peaty soils and peats, respectively, that date to between approximately 8080 and 6740 cal. BP. The uppermost unit comprises peaty lake muds dating from between approximately ad 1790 and the present day (ad 2006). The lower two units contain pollen dominated by forest taxa, whereas the uppermost unit contains pollen indicative of open landscape conditions similar to those of the present. Heavy metal, spheroidal carbonaceous particle, mineral magnetics and stable isotope analysis of the upper sediments show clear evidence of contamination by air pollutants derived from fossil-fuel combustion over the last c. 150years, and diatom analysis indicates that the naturally acidic pool became more acidic during the 20th century. We conclude that the exceptionally acidic surface waters of the pool at present (pH=c. 4.1) are the result of a long history of air pollution and not because of naturally acidic local conditions. We argue that the highly acidic surface waters elsewhere in the NYM are similarly acidified and that the lack of evidence of significant recovery from acidification, despite major reductions in the emissions of acidic gases that have taken place over the last c. 30years, indicates the continuing influence of pollutant sulphur stored in catchment peats, a legacy of over 150years of acid deposition.

Environmental filtering vs. resource-based niche partitioning in diverse soil animal assemblages

Terrestrial invertebrates constitute most of described animal biodiversity and soil is a major reservoir of this diversity. In the classical attempt to understand the processes supporting biodiversity, ecologists are currently seeking to unravel the differential roles of environmental filtering and competition for resources in niche partitioning processes: these processes are in principle distinct although they may act simultaneously, interact at multiple spatial and temporal scales, and are often confounded in studies of soil communities. We used a novel combination of methods based on stable isotopes and trait analysis to resolve these processes in diverse oribatid mite assemblages at spatial
scales at which competition for resources could in principle be a major driver. We also used a null model approach based on a general neutral model of beta diversity. A large and significant fraction of community variation was explainable in terms of linear and periodic spatial structures in the distribution of organic C, N and soil structure: species were clearly arranged along an environmental, spatially structured gradient. However, competition related trait differences did not map onto the distances separating species along the environmental gradient and neutral models provided a satisfying approximation of beta diversity patterns. The results represent the first robust evidence
that in very diverse soil arthropod assemblages resource-based niche partitioning plays a minor role while environmental filtering remains a fundamental driver of species distribution.