Integrating stomach content and stable isotope analyses to elucidate the feeding habits of non-native sharptooth catfish <Emphasis Type="Italic">Clarias gariepinus</Emphasis>

Sharptooth catfish Clarias gariepinus was introduced into the Eastern Cape Province, South Africa, in 1976 and there are concerns about its possible negative impacts on native biota. This study investigated its trophic impact by examining its feeding habits. Stomach content and stable isotope analyses were compared from three localities—the Great Fish River, Sundays River and Glen Melville Dam. Stomach content analysis indicated a catholic diet dominated by fish particularly in all localities. Spatially, however, the diets revealed differences based on the dominance of macrophytes that were only present in the rivers, and aquatic invertebrates that appeared more diverse within the Great Fish River compared to other localities. By contrast, stable isotopes revealed a more generalised feeding pattern with no clear dominance of particular prey. Stable isotopes further showed that the catfish was a complex predator, with large catfish being top predators whereas smaller size groups appeared to feed lower in the food chain. An ontogenetic shift in diet was evident, with small fish predominantly consuming aquatic invertebrates and shifting towards fish with increasing size. High dietary overlap suggests the potential risk associated catfish feeding, especially the potential of piscivory by small catfish that are more likely to persist in shallow and marginal where endangered indigenous minnows occur. The alteration of environmental conditions, especially flow by inter-basin water transfer (IBWT) schemes, was inferred to have had a probable influence its invasion success. Occurrence of other invaders, which was facilitated by the IBWT together with the catfish, posits the risk of invasion meltdown within the study systems.     

Blood fatty acids indicate inter- and intra-annual variation in the diet of Adélie penguins: Comparison with stomach content and stable isotope analysis

Adélie penguin (Pygoscelis adeliae) diet is an important indicator of prevailing environmental conditions and resource availability. In this study, dietary variation within and between years was studied with fatty acid signature analysis (FASA), stomach content analysis (SCA) and stable isotope analysis (SIA). We profiled the fatty acid (FA) composition of whole blood collected from adult penguins throughout the breeding season, and from chicks during the crèche period, in 2001 and 2002. Differences were detected in FA profiles between years, breeding stage and age (adults vs. chicks). These patterns broadly corresponded to those observed from SCA and SIA, with a mix of krill and fish consumed in the early part of the breeding season in both years, krill dominating the diet during the chick-rearing periods in 2001, and fish in 2002. Different metabolic and physiological demands between stages, and ages, may also influence FA profiles but warrants further investigation. In-situ calibrations of adult FA blood profiles were made using corresponding stomach samples to quantify diet composition. Using linear discriminate function analysis, we classified adult FA profiles into 3 meal-types: krill, fish or mixed. A higher proportion of adults had fish-like profiles during the arrival and guard periods. Krill-like profiles dominated during the incubation and crèche periods, although there were a relatively high proportion of fish-like and mixed profiles as well. These patterns corresponded to results from SCA and SIA. This study demonstrates that FASA has the potential to be integrated with other dietary tools to enhance diet monitoring studies, which are currently integral to ecosystem management and conservation measures. The in-situ calibration method used offers a simple and effective alternative to more rigorous calibration techniques developed elsewhere.     

Differential leucocyte cell counts from the pygoscelid penguins of Antarctica

Differential leucocyte counts were obtained for three cogeneric species of wild antarctic penguins, Pygoscelis adelie (adelie), Pygoscelis papua (gentoo), and Pygoscelis antarctica (chinstrap). Significant differences between the differential leucocyte counts of the three species were not observed.     

Tissue turnover and stable isotope clocks to quantify resource shifts in anadromous rainbow trout

Stable isotopes can illuminate resource usage by organisms, but effective interpretation is predicated on laboratory validation. Here we develop stable isotope clocks to track resource shifts in anadromous rainbow trout (Oncorhynchus mykiss). We used a diet-switch experiment and model fitting to quantify N stable isotope (δ¹⁵N) turnover rates and discrimination factors for seven tissues: plasma, liver, fin, mucus, red blood cells, muscle, and scales. Among tissues, diet-tissue δ¹⁵N discrimination factors ranged from 1.3 to 3.4 ‰. Model-supported tissue turnover half-lives ranged from 9.0 (fin) to 27.7 (scale) days. We evaluated six tissue turnover models using Akaike’s information criterion corrected for small sample sizes. The use of equilibrium tissue values was supported in all tissues and two-compartment models were supported in plasma, liver, and mucus. Using parameter estimates and their uncertainty we developed stable isotope clocks to estimate the time since resource shifts. Longer turnover tissues provided accurate estimates of time since resource switch for durations approximately twice their half-life. Faster turnover tissues provided even higher precision estimates, but only within their half-life post-switch. Averaging estimates of time since resource shift from multiple tissues provided the highest precision estimates of time since resource shift for the longest duration (up to 64 days). This study therefore provides insight into physiological processes that underpin stable isotope patterns, explicitly tests alternative models, and quantifies key parameters that are the foundation of field-based stable isotope analysis.     

Evaluating the utility of stable isotope analyses of archived freshwater sample materials

We evaluated the potential utility of stable isotope analysis of tissues commonly archived by aquatic biologists. Previous studies with chemically preserved samples have shown contradictory results, which present an obstacle for the use of archived sample materials. We tested the effects of ethanol and formalin preservation on zooplankton and of ethanol on benthic macroinvertebrate δ¹³C and δ¹⁵N values. We found that neither formalin nor ethanol had a significant effect on δ¹³C and δ¹⁵N values of preserved zooplankton. Nor did ethanol significantly affect δ¹³C or δ¹⁵N values of macroinvertebrates. However, ethanol preservation slightly, but significantly decreased C:N ratios of both zooplankton and macroinvertebrates, probably reflecting some extraction of lipids. Overall, the effects of preservatives on δ¹³C and δ¹⁵N values that we observed were minor. We also compared δ¹³C and δ¹⁵N values analysed from roach scales and perch operculum bones with those analysed from muscle tissue. Decalcification of scales and operculum bones only slightly improved our comparison to muscle tissue δ¹³C and δ¹⁵N values. Decalcified scales had slightly higher δ¹³C and lower δ¹⁵N values. Similarly, decalcified operculum bones showed slightly increased δ¹³C and decreased δ¹⁵N values to those for fish muscle. Our results confirm that scales and operculum bones can provide a suitable proxy for fish muscle in isotope studies with minor correction. We conclude that various archived sample materials can indeed be used with confidence for historical reconstructions of freshwater food webs by stable isotope analysis. Handling editor: K. Martens     

The use of stable isotope ratios in whitebait otolith carbonate to identify the source of prey for Western Australian penguins

A methodology has been developed and used to investigate a direct link between juvenile whitebait (Hyperlophus vittatus), the key prey species sampled from the stomachs of Little Penguins (Eudyptula minor), and the specific nursery area of that prey. A unique application of existing methodology initially involved the measurement of the stable isotopes of oxygen (δ¹⁸O:δ¹⁶O) and carbon (δ¹³C:δ¹²C) in sagittal otolith carbonate, using standard mass spectrometric techniques. The results indicated that the inshore distribution of juvenile (0+ year old) whitebait between Cockburn Sound and Koombana Bay, Western Australia consisted of a number of separate assemblages. Measured differences in stable oxygen isotope ratios were attributed to variations in freshwater input to the embayments that provided the whitebait habitats. In contrast, the measured stable carbon isotope ratios probably resulted from the different isotopic compositions of the food webs in the various habitats. Secondly, a comparison of the average value of carbon and oxygen isotope signatures of pooled otoliths from samples of whitebait from a number of different nearshore coastal sites (assemblages), with that of whitebait obtained from the stomachs of penguins at their main breeding site (Penguin Island) indicated that the values from the penguins resemble most closely those of the average otolith values obtained from whitebait from only one site (Becher Point). Assuming that the whitebait sampled were representative of the whitebait in the nearshore habitats and the diets of the penguins, then these results imply that at the time of sampling the penguins were feeding on whitebait from only one site.     

Food habits of the farmer damselfish <Emphasis Type=Italic>Stegastes nigricans</Emphasis> inferred by stomach content,stable isotope,and fatty acid composition analyses

The territorial damselfish, Stegastes nigricans, maintains algal farms by excluding invading herbivores and weeding unpalatable algae from its territories. In Okinawa, Japan, S. nigricans farms are exclusively dominated by Polysiphonia sp., a highly digestible filamentous rhodophyte. This study was aimed at determining the diet of S. nigricans in Okinawa and its dependency on these almost-monoculture algal farms based on stomach content and chemical analyses. Stomach content analyses revealed that all available food items in the algal farms (i.e., algae, benthic animal inhabitants, trapped detritus) were contained in fish stomachs, but amorphous organic matter accounted for 68% of the contents. Therefore, carbon and nitrogen stable isotope ratios and fatty acid (FA) compositions were analyzed to trace items actually assimilated in their bodies. Stable isotope analyses showed that benthic animals were an important food source even for this farmer fish. Two essential fatty acids (EFAs), 20:4n6 and 20:5n3, which are produced only by rhodophytes among available food items, were rich in the muscle tissue of S. nigricans as well as in algal mats and detritus, suggesting that algal mats contribute EFAs to S. nigricans directly and indirectly through the food web. In conclusion, S. nigricans ingested algal mats, detritus, and benthic animals maintained within its farm. Algae and detritus were original sources of EFAs, and benthic animals, which were much more abundant in the farms than in outside territories, provided a nitrogen-rich dietary source for the fish.     

Ferox Trout (Salmo trutta) as `Russian dolls': complementary gut content and stable isotope analyses of the Loch Ness foodweb

1. Conventional collection methods for pelagic fish species (netting, trawling) are impractical or prohibited in Loch Ness, U.K. To investigate trophic relationships at the top of the Loch Ness food web, an alternative strategy, angling, provided samples of the top predator, the purely piscivorous ferox trout ( Salmo trutta ).
2. The gut contents of these fish provided further samples of prey-fish, and subsequent examination of prey-fish guts revealed their dietary intake, analogous to the famous nested `Russian dolls'. Each trophic level separated by gut content analysis provided further complementary samples for stable isotope analysis and thus information on the longer term, assimilated diet.
3. Ferox trout exhibited considerable cannibalism to supplement a diet of Arctic charr ( Salvelinus alpinus ). However, conspecifics stemmed from a lower isotopic baseline in relation to charr, so ferox trout exhibited a lower trophic level than predicted (4.3) by using the δ¹⁵N values. Charr displayed dietary specialisation with increasing length, and isotopic values supported by the gut data placed the charr at a trophic level of 3.5. The isotope data also indicated that charr carbon was primarily autochthonous in origin.     

Conservation of biotrophy in Hygrophoraceae inferred from combined stable isotope and phylogenetic analyses

The nutritional modes of genera in Hygrophoraceae (Basidiomycota: Agaricales), apart from the ectomycorrhizal Hygrophorus and lichen-forming taxa, are uncertain. New δ(15)N and δ(13)C values were obtained from 15 taxa under Hygrophoraceae collected in central Massachusetts and combined with isotopic datasets from five prior studies including a further 12 species using a data standardization method to allow cross-site comparison. Based on these data, we inferred the probable nutritional modes for species of Hygrophorus, Hygrocybe, Humidicutis, Cuphophyllus and Gliophorus. A phylogeny of Hygrophoraceae was constructed by maximum likelihood analysis of nuclear ribosomal 28S and 5.8S sequences and standardized δ(15)N and δ(13)C values were used for parsimony optimization on this phylogeny. Our results supported a mode of biotrophy in Hygrocybe, Humidicutis, Cuphophyllus and Gliophorus quantitatively unlike that in more than 450 other fungal taxa sampled in the present and prior studies. Parsimony optimization of stable isotope data suggests moderate conservation of nutritional strategies in Hygrophoraceae and a single switch to a predominantly ectomycorrhizal life strategy in the lineage leading to Hygrophorus. We conclude that Hygrophoraceae of previously unknown nutritional status are unlikely to be saprotrophs and are probably in symbiosis with bryophytes or other understory plants.     

Linking aboveground and belowground food webs through carbon and nitrogen stable isotope analyses

Carbon and nitrogen stable isotope ratios (δ¹³C and δ¹⁵N) have been used for more than two decades in analyses of food web structure. The utility of isotope ratio measurements is based on the observation that consumer δ¹³C values are similar (<1‰ difference) to those of their diet, while consumer δ¹⁵N values are about 3‰ higher than those of their diet. The technique has been applied most often to aquatic and aboveground terrestrial food webs. However, few isotope studies have examined terrestrial food web structure that includes both above- and belowground (detrital) components. Here, we review factors that may influence isotopic signatures of terrestrial consumers in above- and belowground systems. In particular, we emphasize variations in δ¹³C and δ¹⁵N in belowground systems, e.g., enrichment of ¹³C and ¹⁵N in soil organic matter (likely related to soil microbial metabolism). These enrichments should be associated with the high ¹³C (~3‰) enrichment in belowground consumers relative to litter and soil organic matter and with the large variation in δ¹⁵N (~6‰) of the consumers. Because such enrichment and variation are much greater than the trophic enrichment generally used to estimate consumer trophic positions, and because many general predators are considered dependent on energy and material flows from belowground, the isotopic variation in belowground systems should be taken into account in δ¹³C and δ¹⁵N analyses of terrestrial food webs. Meanwhile, by measuring the δ¹³C of key predators, the linkage between above- and belowground systems could be estimated based on observed differences in δ¹³C of primary producers, detritivores and predators. Furthermore, radiocarbon (¹⁴C) measurements will allow the direct estimation of the dependence of predators on the belowground systems.     

Gut content and stable isotope analyses provide complementary understanding of ontogenetic dietary shifts and trophic relationships among fishes in a tropical river

1. Despite widespread recognition of the role of body size in fish trophic ecology, little attention has been focused on this issue in isotopic studies, particularly in tropical systems. 2. We used analyses of stomach contents and stable isotopes to examine size‐related shifts in diet in a terapontid fish assemblage in the Australian wet–dry tropics. Stomach content analysis identified substantial ontogenetic dietary shifts in all species, corresponding to changes in body size–isotope trajectories for two species. Shifts away from relatively specialised diets of heavily ¹³C‐depleted insect larvae to consumption of a range of items across multiple basal carbon sources appeared to be the proximate cause of observed isotopic changes. 3. Allochthonous organic matter in the form of C3 riparian vegetation was particularly important to smaller terapontids before larger fish shifted to a broad range of dietary items and similarly broad range of basal carbon sources. 4. While there was general agreement between δ¹³C and stomach content analysis, there was minimal concurrence between the latter and δ¹⁵N isotopic derivation of estimates of trophic position. Due to factors such as omnivory, isotopically overlapping basal sources and uncertainties about rates of isotopic fractionation in both predator and prey species, stomach content analysis provides an essential complement to isotopic methodologies in tropical systems. 5. Given that basal sources supporting any individual species can change markedly with ontogeny, consideration of intraspecific, size‐related variation is necessary in isotopic studies of food web structure.     

定量评价天敌昆虫控害功能的稳定同位素方法

天敌在区域性多作物农田景观系统中辗转捕食害虫,对调节和控制害虫种群数量发挥着非常重要的作用。明确天敌昆虫的食物来源和扩散规律是定量评价其控害功能的重要环节。其中,稳定同位素标记方法是追溯生物个体的食物来源和探究其运动规律的重要手段。本文首先介绍该标记方法的基本原理,接着以天敌昆虫龟纹瓢虫Propylea japonica(Thunberg)为例,应用稳定同位素碳标记方法追溯其在棉花和玉米农田景观系统中取食来源与食物比例;最后叙述了稳定同位素方法的应用前景和存在的问题。     

Diets of crustacean zooplankton,inferred from stable carbon and nitrogen isotope analyses,in lakes with varying allochthonous dissolved organic carbon content

We used stable carbon and nitrogen isotope analyses to estimate the relative proportions of three putative food sources (1) algae, (2) allochthonous organic matter (but including also heterotrophic bacteria and green-sulphur bacteria having similar isotopic values) and (3) methane-oxidizing bacteria (MOB) in the diets of crustacean zooplankton in five small boreal lakes representing a gradient of dissolved organic carbon (DOC) concentration from ca. 5 to 40 mg C l⁻¹. The lakes were sampled in May, after establishment of stratification, and again in October during autumnal mixing of the water column. IsoSource mixing model outputs indicated that the proportion of algae in the diets of zooplankton was generally higher in May than in October, and that bacteria contributed to the diets of both cladocerans and copepods. Our results indicate that bacteria, especially MOB, can make an appreciable contribution to zooplankton diets in these small lakes, even in those with relatively low DOC concentrations.     

Getting to the fat of the matter: models,methods and assumptions for dealing with lipids in stable isotope analyses

Within an organism, lipids are depleted in ¹³C relative to proteins and carbohydrates (more negative δ¹³C), and variation in lipid content among organisms or among tissue types has the potential to introduce considerable bias into stable isotope analyses that use δ¹³C. Despite the potential for introduced error, there is no consensus on the need to account for lipids in stable isotope analyses. Here we address two questions: (1) If and when is it important to account for the effects of variation in lipid content on δ¹³C? (2) If it is important, which method(s) are reliable and robust for dealing with lipid variation? We evaluated the reliability of direct chemical extraction, which physically removes lipids from samples, and mathematical normalization, which uses the carbon-to-nitrogen (C:N) ratio of a sample to normalize δ¹³C after analysis by measuring the lipid content, the C:N ratio, and the effect of lipid content on δ¹³C (Δδ¹³C) of plants and animals with a wide range of lipid contents. For animals, we found strong relationships between C:N and lipid content, between lipid content and Δδ¹³C, and between C:N and Δδ¹³C. For plants, C:N was not a good predictor of lipid content or Δδ¹³C, but we found a strong relationship between carbon content and lipid content, lipid content and Δδ¹³C, and between and carbon content and Δδ¹³C. Our results indicate that lipid extraction or normalization is most important when lipid content is variable among consumers of interest or between consumers and end members, and when differences in δ¹³C between end members is <10–12‰. The vast majority of studies using natural variation in δ¹³C fall within these criteria. Both direct lipid extraction and mathematical normalization reduce biases in δ¹³C, but mathematical normalization simplifies sample preparation and better preserves the integrity of samples for δ¹⁵N analysis.     

Seasonal changes in the diets of migrant and non-migrant nectarivorous bats as revealed by carbon stable isotope analysis

Three species of nectar-feeding bats migrate from tropical and subtropical Mexico into the Sonoran and Chihuahuan deserts during the spring and summer months. We examined geographic and seasonal changes in the diet of one migrant species, Leptonycteris curasoae, using carbon stable isotope techniques to determine the relative importance of C3 and CAM (Cactaceae, Agavaceae) plants in its diet. We also examined the diet of a non-migratory nectar-feeding bat, Glossophaga soricina, from southern Mexico using the same techniques. We found that L. curasoae feeds extensively or exclusively on CAM plants during migration and in the northern part of its range and feeds mostly on C3 plants in southern Mexico. This bat is a year-round resident on Baja California where it is a CAM specialist. The non-migrant G. soricina feeds mostly on C3 plants year-round. Phenological data suggest that certain species of columnar cacti and at least one group of paniculate Agaves on the Mexican mainland provide a spatio-temporally predictable nectar corridor along which nectarivorous bats may migrate in the spring and fall, respectively. Different flowering schedules of Agaves in Baja California appear to promote year-round dietary specialization and perhaps non-migratory behavior in nectar-feeding bats living there.