Multielement isotope analysis of bovine muscle for determination of international geographical origin of meat

Multielemental (C, N, H, S) stable isotope ratio analysis was used as an analytical tool to verify the geographical origin of beef from several European and non-European countries. Beef samples were collected from nine different countries, and the (13)C/(12)C, (15)N/(14)N, (2)H/(1)H, and (34)S/(32)S ratios of defatted beef were measured using isotope ratio mass spectrometry (IRMS). There were highly significant differences in the mean isotopic values of the beef from different countries. The results of discriminant analysis showed that the four isotope ratios were significant for the discrimination of geographical origin and that 84.9% of the samples were correctly assigned to the country of origin (82.2% when cross-validated). Beef was also classified according to geographical origin when additional information on different feeding regimens used in Ireland was included, with 85.0% of the samples correctly allocated and 82.9% cross-validated using the isotopic signatures. All of the Irish beef samples verifiable as pasture-fed beef were correctly classified and then cross-validated.     

稳定性同位素碳、氮在牛不同组织中的变化规律

稳定同位素分析是追溯食品来源的一种有效技术。本文主要探讨了在相同的饲养条件下,牛不同组织中碳、氮同位素组成的变化情况以及品种对其的影响。利用同位素比率质谱仪测定了不同部位脱脂牛肉、不同部位粗脂肪、牛尾毛、血液、肝脏及饲料中的δ13C值和δ15N值,并进行了统计分析。结果表明,品种因素不影响组织中的碳、氮同位素组成;肌肉不同部位、血液、肝脏、牛尾毛之间的δ13C值无显著差异,而肝脏中的δ15N值极显著地高于其它组织;不同部位粗脂肪间的δ13C值也无显著差异。结果说明稳定性同位素碳、氮在牛不同组织中的分馏效应是不一致的。     

Effect of diet on the deposition of n-3 fatty acids, conjugated linoleic and C18:1trans fatty acid isomers in muscle lipids of German Holstein bulls

This study examined the effects of feeding diets rich in either n-3 or n-6 polyunsaturated fatty acids (PUFA) on the fatty acid composition of longissimus muscle in beef bulls. Thirty-three German Holstein bulls were randomly allocated to either an indoor concentrate system or periods of pasture feeding (160 days) followed by a finishing period on a concentrate containing linseed to enhance the contents of n-3 PUFA and conjugated linoleic acids (CLA) in beef muscle. The relative proportion and concentration (mg/100 g fresh muscle) of n-3 fatty acids in the phospholipid and triglyceride fractions were significantly increased (p < or = 0.05) in muscle lipids of pasture-fed bulls. The pasture feeding affected the distribution of individual CLA isomers in the muscle lipids. The proportion of the most prominent isomer, CLA cis-9,trans-11, was decreased from 73.5 to 65.0% of total CLA in bulls fed on concentrate as compared to pasture. The second most abundant CLA isomers were CLA trans-7,cis-9 and CLA trans-11,cis-13 in bulls fed on concentrate and pasture, respectively. Diet had no effect on the concentration of C18:1 trans-11. In contrast, the concentration of the C18:1 trans-13/14, trans-15, and trans-16 isomers in the muscle lipids was up to two times higher in pasture-fed as compared to concentrate-fed bulls. Pasture feeding enhanced the concentration of n-3 fatty acids, but the diet had no effect on the concentration of CLA cis-9,trans-11.     

陕西关中地区肉牛产地同位素溯源技术初探

为应用牛组织中C、N同位素组成的区域分布和同位素指标溯源肉牛产地,本文利用同位素比率质谱仪(IRMS)对陕西关中不同区县来源的牛尾毛样品的1δ3C和1δ5N值进行检测,通过聚类分析,研究陕西关中地区肉牛组织中同位素组成的区域分布情况。结果表明,杨凌区、眉县牛尾毛样品聚为一类;乾县、永寿县样品聚为一类;麟游县样品聚为一类;扶风县、岐山县和凤翔县分类不十分明显,分散在上述3类中。说明陕西关中不同地区肉牛组织中同位素组成存在差异,利用它们可进行肉牛地域的小范围溯源。     

肉牛甲烷排放影响因素的试验研究

为考察不同粗饲料类型、日粮精粗饲料比、能量摄入水平等因素对肉牛甲烷排放量的影响，从而为控制甲烷排放提供理论依据，本研究选用4头18月龄的健康肉用公牛(鲁西黄牛)，针对不同粗饲料类型、饲料精粗比和日粮能量摄入水平进行了试验，采用SF₆示踪法对甲烷排放量进行了测定。试验结果表明：SF₆示踪法可用来快速测定反刍动物的甲烷排放量；影响肉牛甲烷排放量因素的主次顺序依次为：饲料粗料类型＞饲料精粗比＞能量摄入水平；对肉牛饲喂紫花苜蓿和青贮玉米秸比饲喂稻草和干玉米秸时甲烷排放量要小；日粮中粗饲料越多，则肉牛的甲烷排放量越大；随着能量摄入水平的加大，肉牛的甲烷排放量也呈增加的趋势。     

Analysis of fatty aldehyde composition, including 12-methyltridecanal, in plasmalogens from longissimus muscle of concentrate- and pasture-fed bulls

In a large study, 64 German Holstein and German Simmental bulls were randomly allocated to either an indoor concentrate system or periods of pasture feeding followed by a finishing period on a concentrate containing linseed to enhance the contents of beneficial fatty acids in beef. This paper reports the diet effects on the concentration of 12-methyltridecanal (12-MT) and further fatty aldehydes released from plasmalogens in the phospholipids of longissimus muscle of the bulls. Because of the trace level of the important odorant 12-MT in beef, the determination of fatty aldehydes in phospholipids was done by acidic hydrolysis and the reaction of the aldehydes with 2,4-DNPH followed by high-pressure liquid chromatography (HPLC) analysis. The diet affected the 12-MT concentrations in the muscle phospholipids of both breeds. Pasture feeding significantly increased the 12-MT concentrations up to 350 microg/100 g fresh muscle in the muscle phospholipids of German Holstein and German Simmental bulls as compared with the concentrate-fed bulls. Furthermore, pasture feeding resulted in a significant increase of n-octadecanal in the muscle phospholipids of both breeds up to 39.5 mg/100 g fresh muscle. The concentration of n-hexadecanal was not affected by the diet. Pasture feeding as compared to concentrate feeding significantly decreased the concentration of n-octadec-9-enal in the muscle phospholipids. Summarizing, pasture feeding increased the 12-MT concentration, which can be associated with meat of more intensive aroma and better taste.     

不同地域牛尾毛中稳定同位素指纹差异分析

利用同位素比率质谱仪测定来自黑龙江省安达市青肯泡乡和火石山乡、内蒙古太仆寺旗、河北省张家口市和三河市、山东省德州市和聊城市7个地域牛尾毛中的δ13C、δ15N和δ2H值,比较分析了不同地域来源牛尾毛中稳定同位素指纹特征差异,探讨不同地域来源牛尾毛中稳定同位素组成差异、影响因素及地域差异的范围,为牛肉及其产品同位素指纹溯源技术在实际中的应用提供指导。各区域牛尾毛中δ13C值表现为:黑龙江省安达市青肯泡乡(-14.83‰±2.12)、河北省三河市(-14.20‰±1.19)、山东省德州市(-14.16‰±2.51)和山东省聊城市(-15.78‰±1.08)>黑龙江省安达市火石山乡(-18.87‰±1.69)、内蒙古太仆寺旗(-19.29‰±2.43)和河北省张家口市(-19.83‰±1.92);δ15 N值为:黑龙江省安达市火石山乡(8.07‰±0.95)>内蒙古太仆寺旗(5.73‰±0.85)、河北省张家口市(6.65‰±1.45)>黑龙江省安达市青肯泡乡(4.60‰±0.41)、河北省三河市(4.24‰±0.75)、山东省德州市(4.18‰±0.96)和山东省聊城市(4.90‰±0.46);δ2H值为:河北省三河市(-87.78‰±1.99)、山东省德州市(-87.91‰±3.19)和山东省聊城市(-89.24‰±2.90)>河北省张家口市(-94.07‰±3.38)>黑龙江省安达市火石山乡(-97.05‰±3.41)、内蒙古太仆寺旗(-97.55‰±2.79)>黑龙江省安达市青肯泡乡(-102.25‰±2.99)。不同地域来源牛尾毛中稳定碳、氮、氢同位素指纹差异及地域差异范围取决于不同地域喂养牛的饲料组成、地域海拔和纬度的差异程度。     

2H/(1)H and (13)C/(12)C isotope ratios of trans-anethole using gas chromatography-isotope ratio mass spectrometry

Authenticity assessment of trans-anethole is deduced from (2)H/(1)H and (13)C/(12)C isotope ratios, determined by gas chromatography-isotope ratio mass spectrometry (GC-IRMS). For that purpose, self-prepared anise and fennel oils, and synthetic and "natural" samples of trans-anethole, as well as commercially available anise and fennel oils have been investigated. Authenticity ranges of (2)H/(1)H and (13)C/(12)C isotope ratios of trans-anethole were defined. Scope and limitations of the applied online GC-IRMS techniques are discussed.     

Characterization of the geographical origin of Pecorino Sardo cheese by casein stable isotope ((13)c/(12)c and (15)n/(14)n) ratios and free amino acid ratios

The stable isotope ratios ((13)C/(12)C and (15)N/(14)N) of casein measured by isotope ratio mass spectrometry (IRMS) and some free amino acid ratios (His/Pro, Ile/Pro, Met/Pro, and Thr/Pro) determined by HPLC in samples of ewes' milk cheese from Sardinia, Sicily, and Apulia were found to be parameters independent of ripening time. Multivariate data treatments performed by applying both unsupervised (principal component analysis and cluster analysis) and supervised [linear discriminant analysis (LDA)] methods revealed good discrimination possibilities for the cheeses according to place of origin. In this respect, particularly significant were the variables Ile/Pro, Thr/Pro, (13)C/(12)C, and (15) N/(14)N ratios on which basis 100% discrimination and classification of the samples by LDA was obtained.     

Application of stable carbon isotope analysis to the detection of testosterone administration to cattle

The use of anabolic substances is prohibited in food-producing animals throughout the European Union. No method is available to reliably detect the misuse of natural hormones in cattle. A method was developed to detect the abuse of testosterone in cattle fattening. Synthesized testosterone is rather depleted in the (13)C/(12)C ratio. Hence, the method is based on gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) analysis of urine. To select testosterone metabolites and endogenous reference compounds (ERC), the concentration of urinary steroids of cattle was investigated. Dehydroepiandrosterone and androst-5ene-3beta,17alpha-diol were chosen as ERCs to show endogenous (13)C/(12)C ratios. Etiocholanolone and 5alpha-androstane-3beta,17alpha-diol were chosen as the most important testosterone metabolites. Other metabolites known from literature like epitestosterone were less promising. In principle, GC/C/IRMS is a nonspecific method because finally carbon dioxide is analyzed. Therefore, a dedicated cleanup procedure for the selected steroids was developed. By means of proposed confidence intervals in the isotopic composition of ERCs and metabolites, the administration of testosterone to cattle could be detected reliably. Differences of up to 11 per thousand on the delta-scale between ERC and testosterone metabolites were found after testosterone administration, whereas endogenous differences did not exceed 2 per thousand.     

Evaluating N/N and C/C isotope ratio analysis to investigate trophic relationships of elaterid larvae (Coleoptera: Elateridae)

Carbon and nitrogen isotope ratios in consumer tissues can be used to analyse the diet and trophic level of soil animals. However, life history traits may significantly influence stable isotope patterns. We evaluated in a series of experiments how stable isotope ratios of carbon (¹³C/¹²C) and nitrogen (¹⁵N/¹⁴N) at natural abundance can be used to study the diet and trophic position of long-lived macro-invertebrates, elaterid larvae, which are major below-ground herbivores. Small, but significant differences in δ¹³C signatures were found between the larvaes’ anterior and posterior body segments, whereas exuvia reflected the body's overall isotopic composition. The species-specific trophic shift (±SE) in δ¹⁵N for Agriotes obscurus and Agriotes sputator (1.62±0.24‰ and 1.08±0.27‰, respectively) was significantly lower than “mean enrichment estimates” reported in the literature, showing the limited applicability of such generalised estimates in studies of invertebrate trophic ecology. To avoid false-positive assignments to two trophic levels due to variation in δ¹⁵N values, a minimum sample size of three and five individuals for A. obscurus and A. sputator, respectively, was needed to reduce this risk to below α=5%. Keeping elaterid larvae for up to 128 days without food did not affect their isotopic signatures, in contrast to previous studies on starving animals. Switching wireworms to isotopically different diets induced changes in their isotopic signatures within 2 weeks. Changes, however, were significant only when the isotopic difference between diets was large. We conclude that experimental studies evaluating how specific life history traits affect stable isotope signatures in consumers have to precede any interpretation of stable isotope data gathered in the field.     

Influence of feeding soybean oil on conjugated linoleic acid content in beef

Forty-eight steers were used to study the influence of feeding soybean oil (SO) on the conjugated linoleic acid (CLA) content of beef. Steers were fed either a control diet containing 954 g/kg of dry matter (DM) corn-based concentrate (CTL) or a control diet supplemented with SO at 20 (SO2) or 40 g/kg (SO4) of diet DM for 105 days. Adipose tissue samples were collected from the M. longissimus dorsi (LD) and from the M. semitendinosus (ST) on days 0 and 63 of the experiment. Adipose and muscle tissue samples were collected from the LD and ST immediately after slaughter. Feeding 40 g/kg of DM as SO increased the proportions of trans-C(18:1) in beef lipid as compared to CTL and SO2 treatments. The C(18:2) cis-9, trans-11 isomer of CLA as a proportion of total fat was not different in adipose and muscle across treatments. Supplementing SO increased C(18:2) trans-10, cis-12 CLA in adipose tissue of the LD. Supplementing high-grain finishing diets with SO is not an effective strategy to enhance the C(18:2) cis-9, trans-11 isomer of CLA in beef.     

Application of multielement stable isotope ratio analysis to the characterization of French, italian, and spanish cheeses

The stable isotope ratios (delta13C, delta15N, and delta34S of casein and delta13C and delta18O of glycerol) measured by IRMS of French, Italian, and Spanish cheeses are presented and discussed. Variability factors such as animal-feeding regimen, geographical origin, and climatic and seasonal conditions were studied to check the possibilities of cheese characterization offered by each isotopic parameter. Delta13C values of both casein and glycerol appeared to be strongly correlated to the amount of maize in the animal diet. Delta15N and delta34S of casein proved to be mostly influenced by the geoclimatic conditions of the area (aridity, closeness to the sea, altitude). Delta18O of glycerol was more dependent on the geographical origin of the cheeses and on climatic/seasonal parameters. By applying a multivariate stepwise canonical discriminant analysis, good discrimination possibilities for the different European cheeses were obtained, confirmed by the classification analysis, when >90% of the samples were correctly reclassified.     

Selenium accumulation in beef: effect of dietary selenium and geographical area of animal origin

Selenium (Se) is an essential nutrient with multiple human health benefits; the single most important dietary source of Se is beef. The Se content of beef varies, and cattle fed a high selenium diet may have Se concentrations in beef that are well above average. Such beef is potentially a unique supplemental source of dietary Se. To examine factors affecting Se accumulation in beef, 16 steers (initial wt 374.4 +/- 33.7 kg) were taken from seleniferous or nonseleniferous areas and fed in a 2 x 2 factorial design with diets high or moderate in Se (11.9 or 0.62 mg Se/kg diet). Diets contained 50% alfalfa, 25% wheat, and 25% corn on a dry matter basis. All dietary Se was from agricultural products, and Se in the high Se diet was primarily from high Se wheat and alfalfa hay. A loin muscle biopsy was taken at the start of the trial to determine initial Se content of beef. Steers were slaughtered after 14 weeks of the trial, and edible carcass (round, sirloin, shoulder clod, and ribeye) and organ samples were collected. Diets did not affect growth or feed intake (P > 0.05), and Se toxicity signs were not observed. Different cuts of meat had similar Se concentrations, and the Se content of all cuts was increased by both high dietary Se and high Se background. Except for liver and kidney, Se in tissues was increased by seleniferous background (P < 0.02) and high dietary Se (P < 0.001). Kidney Se concentrations of animals fed the high Se diet were lowest in animals from seleniferous areas (P = 0.04), suggesting a possible adaptation to the high Se diet. These results demonstrate that cattle fed diets high in Se from agricultural products will accumulate substantial amounts of Se in the beef without developing signs of Se toxicity and that prior Se status regulates Se accumulation in some organs. They further demonstrate that management practices may be altered so as to make beef a significant source of dietary Se.     

Utilizing Composted Beef Cattle Manure and Slaughterhouse Waste as Nitrogen and Phosphorus Fertilizers for Calcareous Soil

Effectively utilizing composts requires that their nitrogen (N) and phosphorus (P) contents be used as fertilizer, but how this is best accomplished is not fully understood. The authors' objective was to quantify N and P availability of a calcareous clay loam soil receiving composts derived from four contrasting beef cattle feedlot feedstocks applied at 50, 150, and 450 mg total P kg^?1 and supplemented twice with fertilizer N for a 42-week greenhouse plant bioassay. Three composted manures from beef cattle fed distinct diets and a composted mix of slaughterhouse and construction waste were applied. Inorganically fertilized and non-amended soils were included as controls. Canola (Brassica napus L.) and pea (Pisum sativum L.) were grown in pots containing 1.5 kg air-dried soil for six alternating 7-week cycles. Soils amended with composted manure from beef cattle fed typical finishing diets had the lowest apparent N recovery (31%) and the greatest soil nitrate after 42 weeks (25 mg N kg^?1). Phosphorus availability was greater with composted manure from beef cattle fed distillers' dried grains than composted manure from beef cattle fed typical finishing diets and a composted mixture of slaughterhouse and construction waste. Apparent P recovery (66%) was greatest from composted manure of beef cattle fed corn (Zea mays L.) distillers' dried grains applied at 50 mg total P kg^?1. Composted manure from beef cattle fed distillers' dried grains had greater P availability than conventional composted beef cattle feedlot manure. Overall, performance of the composted mixture of slaughterhouse and construction waste was similar to the composted beef cattle manures.     

Stable isotopes revisited: Their use and limits for oribatid mite trophic ecology

In this review we summarize our knowledge of using stable isotopes (¹⁵N/¹⁴N, ¹³C/¹²C) to better understand the trophic ecology of oribatid mites. Our aims are (a) to recapitulate the history of stable isotope research in soil animals with a focus on oribatid mites, (b) to present new stable isotope data for oribatid mites and overview the current state of knowledge of oribatid mite trophic niche differentiation, (c) to compile problems and limitations of stable isotope based analyses of trophic relationships and (d) to suggest future challenges, questions and problems that may be solved using stable isotope analyses and other novel techniques for improving our understanding on the trophic ecology of soil invertebrates. We conclude that (1) in addition to ¹⁵N/¹⁴N ratios, ¹³C/¹²C ratios contribute to our understanding of the trophic ecology of oribatid mites, allowing, e.g. separation of lichen- and moss-feeding species, (2) there likely are many lichen but few moss feeding oribatid mite species, (3) oribatid mite species that are endophagous as juveniles are separated by their stable isotope signatures from all other oribatid mite species, (4) fungivorous oribatid mite species cannot be separated further, e.g. the fungal taxa they feed on cannot be delineated. A particular problem in using stable isotope data is the difficulty in determining signatures for basal food resources, since decomposing material, fungi and lichens comprise various components differing in stable isotope signatures; ¹³C/¹²C ratios and potentially other isotopes may help in identifying the role of these resources for decomposer animal nutrition.     

Effect of wheat-based dried distillers' grains with solubles inclusion on barley-based feed chemical profile, energy values, rumen degradation kinetics, and protein supply

The objectives of this study were to determine the effect of replacing the barley grain portion of the diet by wheat-based dried distillers' grains with solubles (wDDGS) at graded levels on feeding value for beef cattle. Two cultivars of barley were mixed with two sources of wDDGS in ratios of 100:0, 75:25, 50:50, and 25:75% (weight DM basis; denoted B0, B25, B50, and B75, respectively). This study revealed that increasing wDDGS inclusion level increased most of the nutritional composition linearly except for starch, which linearly decreased (from 609 to 320 g/kg of DM). Soluble, slowly degradable, and undegradable Cornel Net Carbohydrate and Protein System (CNCPS) protein and carbohydrate fractions linearly increased with increasing wDDGS inclusion level, whereas their rapidly and intermediately degradable fractions decreased. With increasing wDDGS inclusion, the rumen degradation rate of all measured parameters decreased linearly, the extent of degradability of organic matter was not affected, and the extent of CP degradability (g/kg DM) as well as the predicted protein supply in the small intestine and degraded protein balance in the rumen was increased. The inclusion of wDDGS in barley-based diets up to 50% did not alter energy values of the diet. Furthermore, optimum N to energy balance of the feed mixture for microbial growth in the rumen was reached by replacing 25% of barley by wDDGS. Thus, the nutritive value of the barley-based diets is manipulated by including wDDGS, which can be used to overcome the shortcomings of barley-dominated diets for beef cattle.     

How much do soil and water contribute to the composition of meat? A case study: meat from three areas of Argentina

The main goal of this study was to propose a reliable method to verify the geographical origin of meat, establishing the influence of soil and water on its isotopic and elemental composition. Thus, beef meat, soil, and water samples were collected from three major cattle-producing regions of Argentina (Buenos Aires, Córdoba, and Entre Ríos). Multielemental composition was determined on these three matrices by inductively coupled plasma mass spectrometry (ICP-MS), δ(13)C and δ(15)N by isotope-ratio mass spectrometry (IRMS), and the (87)Sr/(86)Sr ratio by thermal ionization mass spectrometry (TIMS). Soil and drinking water samples could be characterized and clearly differentiated by combining the isotopic ratios and elements, demonstrating differences in geology and climatic conditions of three regions. Similarly, meat originating at each sampling area was characterized and differentiated using only five key variables (Rb, Ca/Sr, δ(13)C, δ(15)N, and (87)Sr/(86)Sr). Generalized procrustes analysis (GPA), using the three studied matrices (soil, water, and meat) shows consensus between them and clear differences between studied areas. Furthermore, canonical correlation analysis (CCA) demonstrates significant correlation between the chemical-isotopic profile of meat with those corresponding to both soil and water (r(2) = 0.93, p < 0.001; and r(2) = 0.83, p < 0.001, respectively). So far, there are clear coincidences between the meat fingerprint and those from soil/water where cattle grew, presenting a good method to establish beef provenance. To the authors' knowledge this is the first report linking the influence of soil and water all together on the composition of beef, presenting the basis for the authentication of Argentinean beef, which could be extended to meat from different provenances.     

Use of a rat model to evaluate tall fescue seed infected with introduced strains of Neotyphodium coenophialum

Experimental cultivars of the pasture grass tall fescue are infected with unique strains of the fungal endophyte Neotyphodium coenophialum, which produce low concentrations of ergot alkaloids. A rat model was evaluated as a tool for rapid, initial screening of experimental cultivars considered to be nontoxic. Rats were fed diets that included seed from experimental cultivars of tall fescue with introduced strains of N. coenophialum and a toxic control diet containing seed of the cultivar Kentucky 31 (KY31), with its endemic strain of N. coenophialum. Rats were preconditioned to a nontoxic diet and then fed treatment diets for 13 days with 5 days at thermoneutrality (21 degrees C) followed by 8 days under heat stress (31 degrees C). For most of the 13-day treatment period, rats fed KY31 exhibited depressed daily intake compared to those fed diets of cultivars with introduced endophytes (P < 0.05). In addition, rats fed KY31 exhibited significantly less weight than rats on other diets after heat treatment was imposed. For all initial trials and repeated trials, total intake and total gain calculated at the end of each trial were the most consistent indicators of toxicity.     

Contrasting effects of nitrogen limitation and amino acid imbalance on carbon and nitrogen turnover in three species of Collembola

Soil animal detritivores play an important role in facilitating decomposition processes but little information is available on how the quality of dietary resources affects their stoichiometry of carbon (C) nitrogen (N) and phosphorus (P), and turnover of C and N. This study investigated how a fungal diet, Fusarium culmorum, with a low N content and imbalanced amino acid (AA) composition affected the physiology of three soil-dwelling collembolans (Folsomia candida, Protaphorura fimata and Proisotoma minuta) in comparison to a control diet, Saccharomyces cerevisiae, with a high N content and balanced AA composition. We compared the elemental composition of animals, their growth rates and tissue replacement of C and N. We also measured the individual AA δ¹³C to investigate the extent that Collembola may rely on endogenous sources to compensate for scarcity of essential AAs. The results showed that animal's N content tracked closely the composition of their diets, decreasing from around 10 to 7% N from the high to low N diet. They also had a significant increase of C and a decrease of P. P. fimata was less affected than F. candida and P. minuta. The total incorporation of C and N in the animals due to growth and tissue replacement decreased from 11-17 to 6-12% DM d⁻¹ on the high and low N diet respectively with P. fimata experiencing the smallest change. Essential AAs δ¹³C did not always match perfectly between Collembola species and their diets; particularly on the low N diet. Isotope patterns of AAs indicate that bacteria may have been the alternative source of essential AAs. While the results of this study cannot be extrapolated directly to the dynamics of Collembola populations in the field, they serve to demonstrate their flexibility in adapting physiologically to the temporal and spatial patchiness of the soil environment.