Nitrogen Stable Isotope Technique Research Articles

Trophic transfer of heavy metals across four trophic levels based on muscle tissue residuals: a case study of Dachen Fishing Grounds, the East China Sea.

In this study, we collected 56 species of fishery organisms (including fish, crustaceans, cephalopods, gastropods, and bivalves) from four seasonal survey cruises at the Dachen fishery grounds. We measured the concentrations of seven heavy metals (Cd, Zn, Cu, Pb, Cr, As, and Hg) in these fisheries organisms. We determined their trophic levels using carbon and nitrogen stable isotope techniques. We analyzed the characteristics of heavy metal transfer in the food chain. The results showed significant differences in heavy metal concentrations among different species. Among all biological groups, bivalves and gastropods exhibited higher levels of heavy metal enrichment than other biological groups, while fish had the lowest levels of heavy metal enrichment. Heavy metals exhibited different patterns of nutritional transfer in the food chain. While Hg showed a biomagnification phenomenon in the food chain, it was not significant. Cd, Zn, Cu, Pb, Cr, and As exhibited a trend of biodilution with increasing nutritional levels, except for As, which showed no significant correlation with δ15N.

Seasonal variations of macrozoobenthic community trophic structure in the waters adjacent to the seaweed beds of Dalian Island in the North Yellow Sea.

To investigate the effects of algal detritus export on the trophic structure of macrozoobenthic community in the adjacent benthic habitat during the bloom and decline of macroalgae, we collected macrozoobenthos from the adjacent sea area of Dalian Island in the North Yellow Sea in May (the algal bloom period) and August (the algal decay period) of 2020. We quantifyied the seasonal changes in the trophic structure of macrozoobenthic community by using carbon and nitrogen stable isotope techniques. Results showed that δ13C and δ15N values of macrozoo-benthos in May ranged from -23.14‰ to -14.24‰, 6.21‰ to 12.90‰, respectively, and -22.36‰ to -14.13‰, 5.33‰ to 12.00‰, respectively in August. Results of PERMANOVA analysis showed that δ13C values of macrozoobenthos differed significantly between the two months, while δ15N values were not significantly different. Based on the Euclidean distance, the macrozoobenthic communities in both months could be classified into five trophic functional groups. The trophic levels of macrozoobenthos ranged from 2.00 (Nitidotellina minuta) to 3.97 (Glycera onomichiensis) in May and from 2.00 (N. minuta) to 3.96 (G. onomichiensis) in August. The δ13C range, δ15N range, mean centroid distance, total area and corrected standard ellipse areas which represented community trophic structure indices in August were higher than those in May. Our results indicated that the trophic diversity level and trophic niche width of the macrozoobenthic community in the adjacent sea area of the seaweed bed were higher in the algal decline season.

Trophic niche and potential carbon source of three reef-associated fishes of Zhongjieshan Islands.

To clarify the trophic relationship of important rock fishes, we analyzed trophic niche of three typical rockfish species (Oplegnathus fasciatus, Sebastiscus marmoratus and Conger myriaster) in the Zhongjieshan Islands in summer 2020, based on the carbon and nitrogen stable isotope techniques. We calculated the contributions of major carbon sources [macroalgae, phytoplankton, suspended particulate organic matter (POM) and substrate organic matter (SOM)]. The results showed that: 1) the δ13C values of the three species ranged from -21.44‰ to -15.21‰, with an average value of (-16.85±1.12)‰, while the δ15N values ranged from 8.32‰ to 10.96‰, with an average value of (9.69±0.66)‰. There were significant differences in carbon and nitrogen stable isotopes among the three species. 2) There was small niche overlap between O. fasciatus and S. marmoratus, indicating that the interspecific competition was not intense. There was no overlap between C. myriaster and the first two, indicating feeding differentiation. 3) The total ecotone area, corrected core ecotone area, and food source diversity of C. myriaster were the highest, indicating that it had a more generalized diet and richer food sources. 4) With Mytilus coruscus as a baseline organism, the trophic level of C. myriaster was the highest (3.38), followed by S. marmoratus (3.09), and the trophic level of O. fasciatus was the smallest (3.00). 5) Results of the stable isotope mixture model (SIAR) showed that POM was the main carbon source of the three species, contributing 57.4%, 57.9%, and 92.0% of the total, respectively. In addition, the contribution rate of SOM was also high for O. fasciatus and S. marmoratus, which was 21.5% and 33.9%, respectively. This study could provide basic information and reference for understanding trophic structure and marine food web in Zhongjiashan Islands.

Stable Isotope Analysis of Food Web Structure and the Contribution of Carbon Sources in the Sea Adjacent to the Miaodao Archipelago (China)

The littoral zones around archipelagos are highly productive coastal habitats that serve as biodiversity hotspots and provide valuable ecosystem services that are different from those of the pelagic and profundal zones. The littoral zone has complex basal carbon sources from different primary producers and is an important ocean–land transition area. Macroalgae are the main primary producers of the littoral zone, but their carbon contribution to consumers is rarely studied. Basal carbon sources determine the structure of the food web. In order to determine the contribution of basal carbon sources and the food web structure of the littoral zone, we used carbon and nitrogen stable isotope techniques and a Bayesian mixing model to study the autumn benthic food web in the sea adjacent to the Miaodao Archipelago. The potential carbon sources of the benthic food web biota in the sea adjacent to the Miaodao Archipelago in autumn are mainly algae (including phytoplankton and macroalgae) and SOM, but the contribution of POM is low. Macroalgae may play a more important role in the littoral zone benthic food webs. Although there are certain uncertainties in the model results, invertebrates and fish have obvious differences in their use of carbon sources. The trophic importance of different primary producers varies with functional feeding groups, and the resource utilization of different functional feeding groups may have certain spatial characteristics.

Food Web Trophic Structure at Marine Ranch Sites off the East Coast of Korea

Understanding the trophic ecology of the giant Pacific octopus Enteroctopus dofleini is challenging in developing marine ranches and in reestablishing its regional stocks against the severe stress of fishing. We adopted carbon and nitrogen stable isotope techniques (termed δ13C and δ15N, respectively) to identify the trophic niche (i.e., pathways and positions) of this species systematically in the entire food webs of two marine ranches off the east coast of the Korean peninsula. While a slight spatial shift in the isotopic nestedness of faunal communities was observed, the δ13C and δ15N values of consumers were distinct and separate among functional groups at both ranches. The consumer δ13C values spanned a broad range between pelagic and benthic sources of organic matter, and their δ15N values recorded a stepwise trophic-level enrichment, indicating that suspension feeders and herbivore-deposit feeders served as baselines of pelagic- and benthic-based trophic pathways, respectively. The δ13C values of predators, including E. dofleini, were arrayed between the two primary consumer groups. Neither δ13C nor δ15N values showed any remarkable variations with increasing octopus weight. Dietary mixing-model calculations indicated that E. dofleini is a generalist predator relying on both benthic- and pelagic-affinity prey, similar to some teleost species that consume a diverse spectrum of prey. In contrast, other teleost groups showed prevalent trophic links with either pelagic- or benthic-based pathways. The trophic-level estimations revealed that E. dofleini occupies an intermediate position slightly below the teleosts. A lack of discrete trophic positions between E. dofleini and teleosts seemed to be indicative of the released teleost predation but instead reflects the imposed food competition. Overall, the results demonstrated that despite compositional changes in the taxa constituting individual trophic groups, E. dofleini occupied a very similar trophic niche in both ranching systems. Finally, as extracted from information based on octopus marine ranches launched on natural rocky bottoms, our isotopic evidence provides a greater understanding of the trophic ecology of this octopus species in nearshore natural habitats along the southwestern margin of its distribution range.

The Response of Estuarine Ammonia-Oxidizing Communities to Constant and Fluctuating Salinity Regimes.

Aerobic nitrification is a fundamental nitrogen biogeochemical process that links the oxidation of ammonia to the removal of fixed nitrogen in eutrophicated water bodies. However, in estuarine environments there is an enormous variability of water physicochemical parameters that can affect the ammonia oxidation biological process. For instance, it is known that salinity can affect nitrification performance, yet there is still a lack of information on the ammonia-oxidizing communities behavior facing daily salinity fluctuations. In this work, laboratory experiments using upstream and downstream estuarine sediments were performed to address this missing gap by comparing the effect of daily salinity fluctuations with constant salinity on the activity and diversity of ammonia-oxidizing microorganisms (AOM). Activity and composition of AOM were assessed, respectively by using nitrogen stable isotope technique and 16S rRNA gene metabarcoding analysis. Nitrification activity was negatively affected by daily salinity fluctuations in upstream sediments while no effect was observed in downstream sediments. Constant salinity regime showed clearly higher rates of nitrification in upstream sediments while a similar nitrification performance between the two salinity regimes was registered in the downstream sediments. Results also indicated that daily salinity fluctuation regime had a negative effect on both ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) community’s diversity. Phylogenetically, the estuarine downstream AOM were dominated by AOA (0.92–2.09%) followed by NOB (0.99–2%), and then AOB (0.2–0.32%); whereas NOB dominated estuarine upstream sediment samples (1.4–9.5%), followed by AOA (0.27–0.51%) and AOB (0.01–0.23%). Analysis of variance identified the spatial difference between samples (downstream and upstream) as the main drivers of AOA and AOB diversity. Our study indicates that benthic AOM inhabiting different estuarine sites presented distinct plasticity toward the salinity regimes tested. These findings help to improve our understanding in the dynamics of the nitrogen cycle of estuarine systems by showing the resilience and consequently the impact of different salinity regimes on the diversity and activity of ammonia oxidizer communities.

Diet Survey and Trophic Position of Macrobrachium nipponense in the Food Web of Anzali Wetland

The oriental river shrimp (Macrobrachium nipponense), an exotic species, has been well adapted and dispersed in Iranian freshwater ecosystems. The feeding ecology and trophic level of this species in the Anzali Wetland, southwestern Caspian Sea, was investigated by both a traditional method and the carbon and nitrogen stable isotope technique. The highest feeding index (FI) was seen in mollusca at 35.9%, followed by detritus at 31.3%. While no significant differences were observed in the preference of food items among different seasons. A Bayesian mixing model was used to estimate the contributions of the stomach contents to the isotopic signature of M. nipponense. According to mixing model results, M. nipponense obtain food from gastropod, fish, shrimp (as cannibalism), insecta, other crustacea, aquatic plants, worms, zooplankton, and unrecognizable content (as detritus) with 30.6%, 16.8%, 12.7%, 5.2%, 4.3%, 3.5%, 0.6%, 0.3%, and 25.9%, respectively. M. nipponense showed a high δ15N and its trophic level (TL = 3.38) was at the top of the Anzali Wetland food web, close to commercially important fish species such as pike, Esox Lucius. Being at this trophic level, this shrimp is expected to possess high nutritional quality for human consumption.

The influences of eutrophication for the carbon and nitrogen sources for aquatic consumer communities—A case study in Lake Baiyangdian

富营养化对湖泊生态系统能流和物流的影响具有不确定性，多数研究仅关注富营养化对浅水湖泊食物网结构和功能的影响，而富营养化对消费者群落碳源和氮源的影响及其时空分异特征较少关注.鉴于此，本研究选取华北平原最大的浅水富营养化湖泊——白洋淀为研究区，依据生境理化参数将白洋淀划分为3类生境（生境1（Ⅰ和Ⅱ区）主要遭受上游府河废水排放影响；生境2（Ⅴ、Ⅶ和Ⅷ区）主要遭受水产养殖和生活污水的影响；生境3（Ⅲ、Ⅳ和Ⅵ区）遭受人为干扰较小）.在2018年4月和8月分别收集了浮游生物、底栖生物和鱼类样品，运用碳、氮稳定同位素技术定量估算3类生境中底栖和浮游生物对消费者群落碳源和氮源贡献百分比的时空分异特征；同时收集水体和沉积物样品进行常规理化参数分析，明晰富营养化对白洋淀消费者群落碳源和氮源贡献百分比的影响.结果表明：1）就水体和沉积物理化参数季节变化而言，除pH相对稳定外，化学需氧量（COD）、沉积物氨氮（NH₃-Ns）、总氮（TN）和沉积物总磷（TPs）4月高于8月，而其他理化参数则8月高于4月；就空间分布而言，温度（T）、水深（WD）、溶解氧（DO）和沉积物总碳（TCs）值在生境3中最高，而其他理化参数的值则在生境1中最高；2）就δ¹³C和δ¹⁵N空间分布而言，对于同一群落，δ¹³C和δ¹⁵N在3类生境中富集程度呈现显著差异；就时间分布而言，不同季节消费者群落的δ¹³C值存在显著差异，而δ¹⁵N值未呈现显著差异，且消费者群落的δ¹³C和δ¹⁵N呈显著负相关；3）就消费者营养级的空间分布而言，3类生境存在显著差异，同一消费者营养级在生境1中最高，在生境3中最低；就时间分布而言，消费者营养级未呈现显著差异；4）浮游生物对消费者群落碳源和氮源贡献百分比4月高于8月，生境1高于其他生境；而底栖生物对消费者群落碳源和氮源贡献百分比则8月高于4月，生境3高于其他生境；5）通过相关分析，结果表明白洋淀消费者群落的δ¹³C与总磷（TP）、TCs、沉积物总氮（TNs）、总有机碳（TOCs）、TPs呈现负相关关系；δ¹⁵N值与TP、TCs、TNs、TOCs、TPs呈正相关关系.因此，湖泊富营养化会影响消费者群落的碳源和氮源，进而改变湖泊生态系统的能流和物流.;The effects of eutrophication on the energy flow and material flow for lake ecosystems need to be studied. Most of previous studies showed the impacts of eutrophication on the structure and function of lake food web, while the influences of eutrophication on the nitrogen and carbon sources for consumer communities were limited. In view of this, the largest shallow eutrophic lake in North China Plain—Lake Baiyangdian was selected as the study area. According to the physical-chemical parameters in water and sediment, the Lake Baiyangdian was subdivided into three habitats(Habitat 1 strongly influenced by wastewater discharge(Sites Ⅰ and Ⅱ), Habitat 2 influenced by aquaculture and densely populated villages(Sites Ⅴ, Ⅶ and Ⅷ), and Habitat 3 influenced the least by human disturbances (Sites Ⅲ, Ⅳ and Ⅵ)). In April and August 2018, the samples for pelagic organisms, benthic organisms and fish were collected, respectively. Carbon and nitrogen stable isotope techniques were used to quantitatively calculate the contribution percentage of benthic and pelagic organism for the carbon and nitrogen sources of consumer communities. The results showed that: 1) according to the seasonal variation of water and sediment parameters, except pH, the values of chemical oxygen demand (COD), Ammonia nitrogen in sediments (NH₃-Ns), total nitrogen (TN), and total phosphorus in sediment (TPs) in April were higher than that in August, while the other parameters were existed higher values in August; in terms of spatial distribution, temperature (T), water depth (WD), dissolved oxygen (DO), total carbon in sediments (TCs) were showed the highest values in Habitat 3, while the other parameters were showed the highest values in Habitat 1; 2) According to the spatial distribution of δ¹³C and δ¹⁵N, for the same populations, the degree of enrichment were appeared significantly different at three habitats; while for the seasonal variation, the values of δ¹³C for the consumer communities showed significant difference, while the values of δ¹⁵N without significant difference; furthermore, the δ¹³C and δ¹⁵N of the consumer communities were existed the negatively correlated; 3) According to the spatial distribution of consumer trophic level (TL), the TL was appeared significantly different at three habitats; for the same consumer community, the highest values were appeared in Habitat 1, while the lowest values were appeared in Habitat 3; in terms of the temporal variation, the TL was not existed the significantly difference; 4) For the pelagic organisms, the contribution percentage for the carbon and nitrogen sources of consumer communities were appeared higher values in April, and the highest values were existed in Habitat 1; while for the benthic organisms, the contribution percentage for the carbon and nitrogen sources of consumer communities were appeared higher values in August, and the highest values were existed in Habitat 3; 5) Through correlation analysis, the results showed that the δ¹³C of consumer communities were appeared the negatively correlated with TP, TCs, TN in sediment (TNs), total organic carbon in sediment (TOCs), TPs; while the δ¹⁵N were existed the positive correlation with TP, TCs, TNs, TOCs, TPs. Therefore, the eutrophication can influence the carbon and nitrogen sources for consumer communities, and change the energy flow and material flow for the lake ecosystem.

Applications of nitrogen stable isotope techniques in the study of nitrogen cycling in terrestrial ecosystems

Applications of nitrogen stable isotope techniques in the study of nitrogen cycling in terrestrial ecosystems

Unraveling the feeding dynamics of Chinese mitten crab-based ecosystems using carbon and nitrogen stable isotope techniques

The Chinese mitten crab (CMC) is an economically important species that consumers prefer mainly because of its delightful taste and aroma. The taste of CMC varies depending on its environmental conditions. Consumers prefer lake-sourced crabs to pond-cultured ones, but the production of crabs in lakes is greatly discouraged because of its adverse impacts on the local ecosystem. This study investigates the dynamics of the food web structure and trophic levels (TL) of both lake and pond ecosystems to apply the knowledge of the lake-cultured system to pond-cultured production using the stable isotope ratios of C and N. Furthermore, the TL was estimated and the IsoSource model was used for diet contribution estimates. The results show that the δ13C of the crab in the pond ecosystem (− 22.6 ± 0.3‰) was more enriched than that in the lake ecosystem (− 25.7 ± 0.4‰) indicating a clear distinction (P 0.05), suggesting a similar TL. The crabs also occupied higher TL in both ecosystems under consideration, indicating a reliance of crab on the animal matter for food. This study provides more insights for all concerned stakeholders to make an informed decision with respect to the production and consumption of CMC.

Mercury bioaccumulation in arthropods from typical community habitats in a zinc-smelting area.

This study assessed the enrichment of mercury in the food web from the different community habitats in a zinc-smelting area of China. We used a nitrogen stable isotope technique to analyze trophic level relationships among arthropods and found that the first trophic level consisted of plants in the different community habitats, the second trophic level consisted of herbivores such as locusts and grasshoppers (primary consumers), and the third trophic level included spiders and mantes (secondary consumers). Mercury enrichment in the primary consumers was not evident, but enrichment in arthropods of the third trophic level was significant. The average of enrichment coefficients in spiders and mantes was greater than 1. The δ15N values indicated that mercury concentrations accumulated from primary producers to top carnivorous arthropods increased. In this zinc-smelting area, the biological amplification of mercury in the food web is significant. It is reasonable to assume that humans, located at the top of the food chain, are exposed to biomagnified levels of mercury.

Determination of the in situ growth rate of Microcystis based on carbon and nitrogen stable isotope fractionation

Abstract Stable isotope fractionation of carbon and nitrogen in algal cells can be affected by photosynthesis, temperature, nutrient and CO2 concentrations, and cell size. As a consequence, carbon and nitrogen stable isotope techniques are not popular for determining algal growth rates. To counter these issues, this study used BG11 medium to cultivate Microcystis in the laboratory. First, the carbon and nitrogen stable isotope values of the culture medium and the algae are determined. Then, based on changes in isotope fractionation before and after cell division, a function μ = 1.32(1 + x)−0.52 relating growth rate and stable isotope fractionation is established. By substituting stable isotope values from Taihu Lake water and Microcystis into this function, the growth rate of the Microcystis in Taihu Lake is calculated to be 0.64 d−1 in May and 0.12 d−1 in September, with an average growth rate of 0.42 d−1. By incorporating most of the above-mentioned factors influencing isotope fractionation, this method can determine the growth rate of algae based directly on the stable isotope fractionation relationship, enabling simple and practical determination of algae growth rates.

Isotopic invisibility of protozoan trophic steps in marine food webs

According to modern oceanographic perspectives that emphasize microbial pathways, phagotrophic protists comprise one to several levels of intermediate consumers between phytoplankton and larger metazooplankton (copepods and krill). However, recent attempts to quantify pelagic trophic structure in the open ocean using nitrogen stable isotope techniques have brought into question whether such measurements adequately account for protistan trophic steps. Here, we use a two‐stage chemostat system, with Dunaliella tertiolecta and Oxyrrhis marina as a predator—prey model, to address this question experimentally. To investigate 15N trophic discrimination under different conditions of nitrogen availability and recycling, Oxyrrhis was fed in the light and in the dark on phytoplankton provided with high and low nutrient ratios of N : P. We used both bulk and amino acids—compound specific isotopic analysis (AA—CSIA) to distinguish trophic fractionation from changes in the δ15N values of phytoplankton (isotopic baseline). Results demonstrate that protistan consumers are not, in fact, significantly enriched in 15N relative to their prey, a marked departure from the general findings for metazoan consumers. In addition, we show that changes in the isotopic baseline propagate rapidly through the protistan food chain, highlighting the need to account for this variability at ecologically relevant time scales. If protistan trophic steps are largely invisible or significantly underestimated using nitrogen isotope measurements, research that utilize such measurements in ecological, fisheries, and climate change studies may miss a large part of the ocean's variability in food‐web structure and ecosystem function.

The use of nitrogen-stable isotope technique to identify L-leucine extracted from human hair

The production of L-leucine from pure botanical sources is expensive and time consuming. Hence, most fraudulent manufacturers in China use cheap and sometimes unwholesome sources of raw materials such as human hair and duck feathers in its production. Consequently, major concerns on health, economic, religious, legal and allergic reaction cases have been raised about this economically motivated adulteration. Notably, consumers of dietary supplements across the globe are beginning to request for non-animal and non-human sources. Hence, the prediction of the respective sources is essential for consumer protection. Therefore, this study provides discrimination between leucine derived from human hair and pure botanical sources as well as other sources such as duck feathers and pig fur by using the nitrogen stable isotope technique. An elemental analyzer which was connected to an isotope ratio mass spectrometer operated in the continuous flow mode was utilized. Statistical analysis was performed using descriptive statistics and one-way ANOVA. The results obtained showed δ15N values between 8.24 and 8.46 ‰ for the L-leucine extracted from human hair whereas ranges of 0.82–2.23, 5.41–5.47 and 5.88–6.01 ‰ were recorded for that of maize, pig fur and duck feather samples, respectively. It was shown that there is a clear difference between the δ15N composition of L-leucine extracted from human hair and the other sources. The difference confirms the predictable trophic enrichment of δ15N composition of organisms in a food web. This study adds to the efforts already in place to help curb food safety menace.

Authentication of Pure L‐Leucine Products Manufactured in China by Discriminating between Plant and Animal Sources Using Nitrogen Stable Isotope Technique

L-leucine products among other branched chain amino acid supplements are highly susceptible to economically motivated adulteration. Curbing this menace is critical and timely. Hence, the δ(15) N composition of the L-leucine derived from plants and animals sources was estimated. The trophic enrichment phenomenon of δ(15) N composition was utilized to elucidate the sources. We finally established the distinction between the respective sources. Samples of plant sources (maize and soybean) and that of animal sources (pig fur and duck feather) were analyzed for δ(15) N isotopic signatures. An elemental analyzer which was connected to an isotope ratio mass spectrometer operated in the continuous flow mode was utilized. The raw materials were obtained from China. Statistical analysis was performed using descriptive statistics and one-way analysis of variance. The results indicated lower δ(15) N values of range -0.7344‰ to 2.384‰ and 1.032‰ to 2.064‰ for maize and soybean samples, respectively. Whereas, a range of 3.860‰ to 6.011‰ and 5.875‰ to 6.011‰ was, respectively, detected in pig fur and duck feather samples. The δ(15) N difference in plants and animals samples was significant (F = 165.0; P = 1.675 E-10 for maize and pig fur samples; F = 212.8; P = 0.0001284 for soybean and duck feather samples). It was observed that δ(15) N trophic enrichment is helpful in elucidating the respective sources. The authors can emphatically assert that the range of δ(15) N composition of L-leucine derived from plants sources within the study area is -1.000‰ to 3.000‰ whereas the range in animal sources is 4.000‰ to 9.000‰. Practical Application This study provides a reliable approach in verifying the authenticity of not only L-leucine products but also other branched chain amino acid supplements and thereby would help in fraud detection of any economically motivated adulteration and mislabeling of these products. When coupled with H and O stable isotope techniques, the region-of-origin of the detected adulteration can also be traced successfully. It therefore serves as a guide to food regulatory bodies, food scientists, retailers of these products, consumers, and the general public at large.

Rastreabilidade da farinha de carne e ossos bovinos em ovos de poedeiras alimentadas com ingredientes alternativos

O objetivo deste trabalho foi detectar traços de farinha de carne e ossos bovinos, em ovos de poedeiras alimentadas com dietas comerciais com inclusão de ingredientes vegetais alternativos e leveduras. A detecção foi feita pela técnica dos isótopos estáveis do carbono e do nitrogênio. Foram utilizadas 384 poedeiras, distribuídas aleatoriamente em oito tratamentos. Os tratamentos consistiram de uma dieta-controle - à base de milho e farelo de soja - e sete dietas com inclusão de farinha de carne e ossos bovinos, acrescidas ou não de outros ingredientes (farelo de trigo, quirera de arroz, farelo de algodão, glúten de milho, levedura de cana e levedura de cerveja). No 35º dia, foram tomados aleatoriamente 24 ovos por tratamento: 12 para análise de ovo e 12 para análise de gema e albúmen, em separado. Após análise isotópica de carbono e nitrogênio, os resultados foram submetidos à análise multivariada de variância. As médias dos pares isotópicos dos ovos, gema e albúmen, em todos os tratamentos, diferiram daquelas do tratamento-controle. A técnica dos isótopos estáveis permite detectar, nos ovos, gema e albúmen, a farinha de carne e ossos bovinos utilizada na dieta de poedeiras, mesmo com a inclusão de outros ingredientes vegetais e leveduras.