Carbon and nitrogen isotopic similarity between the endangered Darwin's fox (Lycalopex fulvipes) and sympatric free-ranging dogs in Chiloé Island, Chile.

Darwin's fox is an opportunistic omnivorous predator native to Chile classified as endangered by the IUCN Red List. Habitat use by Darwin's foxes can be negatively affected by the presence of free-ranging dogs that range freely across native and non-native habitats and can be a source of fox mortality. The objective of this study was to analyze the isotopic similarity of Darwin's fox and sympatric free-ranging dogs in Chiloé Island to determine the impact of anthropogenic environmental alterations on wild predators. We use hair samples to characterise and compare their δ13C and δ15N values and to evaluate isotopic similarity and isotope niches overlap. A generalised linear model was used to associate the isotope value with landscape variables (forest cover and vegetation type) and distance to the nearest house. We found no significant differences in δ13C or δ15N values between foxes and dogs, and a marginally significant isotope niche overlap (59.4 %). None of the selected variables at landscape and site scale were related to isotope values. Although our study is not a probe of direct contact between foxes and free-ranging dogs, the high isotopic similarity highlights the risk of pathogen spillover from free-ranging dogs to Darwin's foxes.

Rastreability of bovine meat and bone meal using the stable isotope technique in egg production

The present study aimed to verify whether the inclusion of other ingredients in the diet of laying hens could interfere with the traceability of bovine meat and bone meal (BMBM) in eggs through stable carbon and nitrogen isotope analysis. Two hundred and fifty-six white laying hens were used and randomly distributed into eight treatments composed of: CONTROL: control diet; GLUTEN: control diet + corn gluten; YEAST: control diet + yeast; GLUTEN + YEAST: control diet + corn gluten + yeast; BMBM: control diet + 4.5 % BMBM; BMBM + GLUTEN: control diet + corn gluten + 4.5 % BMBM; BMBM + YEAST: control diet + yeast + 4.5 % BMBM; BMBM + GLUTEN + YEAST: control diet + corn gluten + yeast + 4.5 % BMBM. The isotopic results were subjected to multivariate analysis of variance (MANOVA) and LDA multivariate linear discriminant analysis. At 28 days, the eggs of the birds given diets without the addition of BMBM differed from the CONTROL group. The yolks showed that all treatments were significantly different from the CONTROL, and at 56 days, all eggs and egg yolks were different from the CONTROL. At 28 days, albumen was significantly different for all CONTROL experimental treatments; however, at 56 days, the YEAST treatment showed no difference from the control. Thus, it was concluded that even with the addition of other ingredients, the isotope technique is still able to detect the BMBM in eggs.

Rastreability of bovine meat and bone meal using the stable isotope technique in egg production

ABSTRACT The present study aimed to verify whether the inclusion of other ingredients in the diet of laying hens could interfere with the traceability of bovine meat and bone meal (BMBM) in eggs through stable carbon and nitrogen isotope analysis. Two hundred and fifty-six white laying hens were used and randomly distributed into eight treatments composed of: CONTROL: control diet; GLUTEN: control diet + corn gluten; YEAST: control diet + yeast; GLUTEN + YEAST: control diet + corn gluten + yeast; BMBM: control diet + 4.5 % BMBM; BMBM + GLUTEN: control diet + corn gluten + 4.5 % BMBM; BMBM + YEAST: control diet + yeast + 4.5 % BMBM; BMBM + GLUTEN + YEAST: control diet + corn gluten + yeast + 4.5 % BMBM. The isotopic results were subjected to multivariate analysis of variance (MANOVA) and LDA multivariate linear discriminant analysis. At 28 days, the eggs of the birds given diets without the addition of BMBM differed from the CONTROL group. The yolks showed that all treatments were significantly different from the CONTROL, and at 56 days, all eggs and egg yolks were different from the CONTROL. At 28 days, albumen was significantly different for all CONTROL experimental treatments; however, at 56 days, the YEAST treatment showed no difference from the control. Thus, it was concluded that even with the addition of other ingredients, the isotope technique is still able to detect the BMBM in eggs.

Rastreability of bovine meat and bone meal using the stable isotope technique in egg production

The present study aimed to verify whether the inclusion of other ingredients in the diet of laying hens could interfere with the traceability of bovine meat and bone meal (BMBM) in eggs through stable carbon and nitrogen isotope analysis. Two hundred and fifty-six white laying hens were used and randomly distributed into eight treatments composed of: CONTROL: control diet; GLUTEN: control diet + corn gluten; YEAST: control diet + yeast; GLUTEN + YEAST: control diet + corn gluten + yeast; BMBM: control diet + 4.5 % BMBM; BMBM + GLUTEN: control diet + corn gluten + 4.5 % BMBM; BMBM + YEAST: control diet + yeast + 4.5 % BMBM; BMBM + GLUTEN + YEAST: control diet + corn gluten + yeast + 4.5 % BMBM. The isotopic results were subjected to multivariate analysis of variance (MANOVA) and LDA multivariate linear discriminant analysis. At 28 days, the eggs of the birds given diets without the addition of BMBM differed from the CONTROL group. The yolks showed that all treatments were significantly different from the CONTROL, and at 56 days, all eggs and egg yolks were different from the CONTROL. At 28 days, albumen was significantly different for all CONTROL experimental treatments; however, at 56 days, the YEAST treatment showed no difference from the control. Thus, it was concluded that even with the addition of other ingredients, the isotope technique is still able to detect the BMBM in eggs.(AU)

Microencapsulated and uncoated butyric acid as alternative additives to the regeneration of intestinal mucosa in broilers challenged with Eimeria spp.

1. The effect of microencapsulated and uncoated butyric acid as an alternative to antibiotics on performance, intestinal morphology and regeneration of intestinal mucosa was studied in birds experimentally infected with Eimeria spp. 1 to 42 d-old.2. A total of 1,320 male Cobb® broiler chicks were allocated to one of five treatments in a completely randomised design, comprising a negative control, uncoated butyric acid (UA), microencapsulated butyric acid (MA), combined U + M butyric acid and a positive control (antibiotic+anticoccidial) in six replications. At 16 d-old, the birds were inoculated orally with 0.5 ml of a solution containing an Eimeria spp. pool.3. At 21 d of age, the birds receiving butyric acid alone had higher body weight gain (BWG) and feed intake (FI) compared to those supplemented with the blend of acids. For the total rearing period, in all variables, the positive control performed best (P < 0.001).4. At 14 d of age, birds that received diets containing UA had a deeper crypt depth in the jejunum than those fed diets containing microencapsulated acid (P = 0.0194). At 21 d of age, the birds fed the acids had higher villi (P = 0.0058) in the duodenum, compared to the negative control group.5. Supplementation with microencapsulated acid contributed to the intestinal health and recovery of post-challenge birds, but did not result in improvements in performance.

Isotopic carbon turnover in pig hoof and rib

The objective ofthisstudy was to evaluatethebehavior of carbon incorporation andturnoverin hoof and ribs of pigsat different periodsof development in the search fortissuesthat reflectlongerthe formerdiet.We used132commercial hybrids (barrows and females), weaned atan average age of21 days,distributed ina completely randomized designwithfour treatmentsondifferent daysof substitution of corn(C4cycleplantgrain) dietswithbrokenrice (C3 cycleplantgrain) at 21, 42, 63 and 110 daysof age tochangethe carbon-13 isotope signal.By means ofisotopicdilution curves, we observedthat animalswhose C4dietwasreplaced withC3dietat 21, 42, 63and 110days of age,for hoof and rib,reached anew levelof isotope equilibrium. Bone samplesare better choicesto reflect theformer diet, due to conservation of the isotopic signal for longer.(AU)

Isotopic carbon turnover in pig hoof and rib

The objective ofthisstudy was to evaluatethebehavior of carbon incorporation andturnoverin hoof and ribs of pigsat different periodsof development in the search fortissuesthat reflectlongerthe formerdiet.We used132commercial hybrids (barrows and females), weaned atan average age of21 days,distributed ina completely randomized designwithfour treatmentsondifferent daysof substitution of corn(C4cycleplantgrain) dietswithbrokenrice (C3 cycleplantgrain) at 21, 42, 63 and 110 daysof age tochangethe carbon-13 isotope signal.By means ofisotopicdilution curves, we observedthat animalswhose C4dietwasreplaced withC3dietat 21, 42, 63and 110days of age,for hoof and rib,reached anew levelof isotope equilibrium. Bone samplesare better choicesto reflect theformer diet, due to conservation of the isotopic signal for longer.

Strengths and challenges of δ15N to identify anthropogenic nutrient loading in coastal systems.

Coastal ecosystems are under increasing stress from anthropogenic nutrient loading; which is most often assessed through water quality measurements. Here, 136 published studies on the use of δ15N to identify nutrient loading in coastal systems were analyzed to identify key strengths and challenges when using this isotope technique. δ15N has been used successfully for this purpose around the globe for over 40 years. Studies have mainly used benthic macroalgae and sediment samples in estuaries and coral reefs of North America and Oceania. Strengths of this technique include timely identification of nutrient loading and its sources, even when inputs are pulsed or assimilated by biota, the benefits of varying isotope turnover rates in different types of samples, sporadic sampling efforts, simple collection and preparation of samples, and relatively low analysis costs. The shortcomings of this technique have led to a loss in popularity in recent times, mainly from isotopic overlap of potential sources and the effects of other confounding factors on isotopic compositions. These challenges can be compensated by simultaneous measurement of other key variables including additional isotopes (δ13C, δ34S), water column nutrient concentrations, and fecal coliforms, highlighting great potential to use this tool.

Carbon and nitrogen isotopes to distinguish sources of sedimentary organic matter in a Caribbean estuary.

Carbon and nitrogen stable isotope compositions (δ13C and δ15N) of organic matter (OM) and total organic carbon to total nitrogen ratio (Corg/TN) in a sediment core collected in Sagua estuary (Cuba), were investigated to elucidate the origin of the Sedimentary OM (SOM) and changes in its main sources, over the last 100 years. Results showed almost constant values in the elemental and isotope composition of SOM from 1908 to 1970 with an abrupt change after 1970. From 1970 to 2005, δ13C increased from -21.2 up to -19.3 ‰, while δ15N declined from 1.5 to values close to 0 ‰. The output of the mass-balance model for the identification of OM sources indicated that δ13C and Corg/TN values are generally influenced by marine Particulate OM (POM) sources. Between 1900 and 1970, the main OM source in sediments was marine POM (>85 %), with freshwater POM contributing ca. 15%. Since 1970, the establishment of the Alacranes Dam determined drastic environmental changes influencing the OM sources in the area. Mixing models pointed to seagrasses (79 %) as the main contributors to SOM in the first period, while since 1973 onward, the contribution of human-derived sources such as fertilizers and urban discharges became greater. This information can provide baseline data for the environmental management of the Sagua watershed.

Multi-isotope (δ 2H, δ 18O, δ 13C-TDIC, δ 18O-TDIC, 87Sr/86Sr) and hydrochemical study on fractured-karstic and detritic shallow aquifers in the Pampean region, Argentina.

Fluxes between fractured-karstified and detritic aquifers are commonly poorly understood in many environments. These two types of aquifers are in contact in the southeastern Pampean region in the Argentine Buenos Aires province, and the aim of this work is to analyze their relationship contributing to improve the hydrological model. A joint application of hydrochemical and multi-isotope (δ 2H, δ 18O, δ 13C-TDIC, δ 18O-TDIC, 87Sr/86Sr) tools was used. TDIC, δ 2H, δ 18O and δ 13C-TDIC allowed differentiating two main end members. Water in the Pampeano aquifer (PA) which is transferred from the fractured-karstic aquifer (F-KA) is characterised by high TDIC around 500-700 mg/L, isotopically depleted in 18O (about -5.5 ‰) and high δ 13C-TDIC (around -10.0 ‰). The other end member is direct recharge water infiltrated into the PA with TDIC ranging from 400 to 500 mg/L, slightly enriched in 18O (δ 18O = -4.8 ‰), and δ 13C-TDIC in the range of soil CO2 as a result of reactions with calcrete concretions (from -20.0 to -9.0 ‰). Dolomite dissolution is the main process controlling the chemistry of the low-mineralized (Mg-Ca-HCO3) waters, whereas high-mineralized (Na-HCO3) waters are strongly influenced by ion-exchange reactions with adsorbed Ca2+ and Mg2+ and by evaporation.

Isotope composition of carbon dioxide and methane in a tropical urban atmosphere.

This work presents a weekly carbon isotope composition analysis (June 2017-January 2018) of carbon dioxide (CO2) and methane (CH4) in a tropical urban atmosphere (Central Valley, Costa Rica). δ13C values of CO2 and CH4 ranged from -12.2 to -5.9 ‰, and from -51.6 to -46.3 ‰, respectively. Mixing ratios of CO2 and CH4 varied from 384.2 to 528.5 ppmv, and from 1.860 to 2.613 ppmv, respectively. δ13C spatial variation and mixing ratios of CO2 and CH4 were influenced by the atmospheric stability and air circulation patterns in the metropolitan area. Low δ13C values and large mixing ratios were observed in the southwestern area of the valley during the rainiest period (September-November). Preliminary linear relationships between reciprocal CO2 mixing ratios and δ13C values indicate that CO2 emissions in the Central Valley are probably related to respiration processes and fossil fuel combustion, although CO2 enriched in 13C from volcanic degassing was also detected. Under stable atmospheric conditions, CH4 data seems to reflect the influence of emissions near the sampling sites. These preliminary results based on the carbon isotope technique demonstrate potential for carrying out atmospheric studies at tropical urban locations with different terrain characteristics and atmospheric mixing conditions.

Geochemical evolution, residence times and recharge conditions of the multilayered Tubarão aquifer system (State of São Paulo - Brazil) as indicated by hydrochemical, stable isotope and 14C data.

The Tubarão aquifer system constitutes a very complex, multilayered aquifer enclosed in the Paraná basin (central-southern part of Brazil). Despite the relatively low productivity of wells, groundwater represents an important source of water for the very populated and industrialized zones of the State of São Paulo. An extensive water sampling campaign was carried out followed by hydrochemical and isotopic (δ 2H, δ 18O, δ 13C and 14C) studies, aiming at a better understanding of the aquifer's geochemical evolution, recharge processes, and its groundwater residence times. Two main hydrochemical facies were recognized and divide the aquifer in two portions. The shallow portion - the active hydrological zone of the aquifer - is characterized by the Ca-HCO3 water type, evolving as a system open to atmospheric CO2. Mean residence times are typically lower than 5000 years. The lower portion is mostly characterized by the stagnant, Na-HCO3 water type, evolving under closed system conditions. Residence times average up to 15,000 years, but can reach 44,000 years, which indicates the exploitation of (possibly non-renewable) fossil waters. This study contributes to the establishment of proper policies regarding the sustainable groundwater exploitation of the Tubarão aquifer system.

Identifying the composition of commercial Brazilian cat food with stable isotopes of carbon and nitrogen.

Considering the increasing pet owner's concern about the food their pets are consuming, in this study we investigated the origin of the main ingredients in wet and dry foods produced in Brazil using stable isotope ratios of carbon and nitrogen. We concluded that chicken and pork seem to be the dominant ingredients in most of the samples, with larger proportions in wet cat food. Even in pet foods showing 'beef' as the main ingredient on the label, we found a low proportion of bovine products in both wet and dry cat foods. Comparing the contribution of plant-derived products (C3 and C4 plants) and animal-derived products (chicken-pork, bovine and fish), approximately 21 % of cat foods had more than 30 % of ingredients with plant origin in their composition. The high amount of plant-derived products in cat foods found here raises the question whether this should be mentioned on package labels.

Soluble melanins of the Randia echinocarpa fruit - Structural characteristics and toxicity.

The water-soluble melanins (SM) of Randia echinocarpa fruit possess interesting biological activities and have been scarcely characterized. In this study, SM were obtained at boiling (SMBT) and room (SMRT) temperatures and characterized by UV-Vis, IR, thermogravimetric analysis, and GC-MS of the hydrolysis products of the SM; besides, the solid-state 13 C NMR, elemental analysis, and acute and sub-acute toxicity of the SMBT were determined. SMBT and SMRT contain organic acids and carbohydrates and their spectroscopic signals and thermograms were similar, but the SMBT yield was higher. The SMBT were characterized by their elemental composition (C 48.260 ± 0.011%, N 3.693 ± 0.009% and H 6.093 ± 0.076%) consistent with the presence of aromatic rings and eumelanins, degradation temperature at 300°C, 13 C NMR signals supporting melanin-bonding with carbohydrates and organic acids, and innocuity in Balb/C mice (acute assay, LD50  > 5 g/kg b.w.; sub-acute assay, no lethality at 500 mg/kg b.w. for 30 days). PRACTICAL APPLICATIONS: The consumption of melanins has been associated with health benefits because of their biological activities (e.g., antioxidant, immunostimulatory, UV- and radiation-protective). Randia echinocarpa is employed in Mexican traditional medicine against chronic degenerative diseases (e.g., cancer and diabetes) and ailments of organs (e.g., kidney and lung) and systems (e.g., circulatory and gastrointestinal). The R. echinocarpa fruit contains water-soluble melanins (SM) that inhibit carbohydrate-digestive enzymes and show high antioxidant activity; thus, SM could be useful for the prevention and treatment of diabetes. This study showed that the SM structure contains melanin-bonding organic acids and carbohydrates, which could be associated with the SM solubility and higher yield, and that SMBT were innocuous in the acute and sub-acute assays in mice. Thus, the R. echinocarpa SMBT could be used as safe potential ingredients to develop functional products.

Stable isotope analysis of white-tailed deer teeth as a paleoenvironmental proxy at the Maya site of La Joyanca, northwestern Petén, Guatemala.

Carbon and oxygen isotopes ratios from herbivore teeth have previously been used as paleo-environmental proxies in temperate zones. However, their utility in tropical zones remains uncertain. In this study, sequential sub-samples from white-tailed deer (Odocoileus virginianus) teeth (second and third molars) from the Maya archaeological site of La Joyanca, located in northwestern Petén, Guatemala, show that δ18O of enamel carbonate corresponds broadly to modern observed precipitation δ18O over the 10-month period of tooth formation, capturing rainfall seasonality. The analyses also detect significant diachronic differences in the δ18O between the periods 1100-1000 BP (850-950 A.D.) and 1000-900 BP (950-1050 A.D.) at La Joyanca. The δ13C in both periods are indicative of a C3-plant based diet, which suggests cultivation of maize did not differentially affect deer diet during this period.

Does trawl fisheries affect the diet of fishes? A stable isotope analysis approach.

Trawling is the main fishing practice worldwide and its ecosystem effects have been raising concern over the past decades. Long-term impacts can be monitored through changes in the trophic structure, and several studies evaluated trophic level (TL) shifts in fish populations between trawled and untrawled environments. However, published results are contrasting. We performed a metanalysis integrating all available studies that evaluated TL shifts in fishes between trawled and untrawled environments and conducted a local study comparing several features of the trophic ecology in two species of fishes. According to the metanalysis, TL does not change significantly with trawling. In contrast, the local study showed higher TLs and broader isotopic niches in the trawled environment. Diet reconstruction indicated a potential consumption of hake, the main discard component, at the trawled environment. All the studies used in the metanalysis were conducted in the Northern Hemisphere, whereas the local study represents the first data available from the Southern Hemisphere. As industrial commercial fisheries in Argentina are relatively recent, it is possible that our data are capturing the initial stage of ecological changes induced by trawling, compared with the historical fisheries located at the Northern Hemisphere.

Is the 'canine surrogacy approach' (CSA) still valid for dogs and humans in market-oriented and subsistence-oriented communities in Brazil?

Based on the assumptions that human food is available for dogs and isotope diet-tissue differences are similar in dogs and humans, the 'canine surrogacy approach' (CSA) has been used to infer patterns of ancient populations. The goal of this study was to test the CSA in urban (Brasília and Piracicaba) and in rural (Ubatuba and Maraã) areas. The hair C and N isotope ratios of modern dogs were compared with those of human fingernails from different regions of Brazil. Our CSA results showed a correlation between dog and human isotopes values: in rural areas δ15N of humans and dogs was not statistically different; contrarily, in urban centres, δ15N of humans was approximately 1 ‰ higher (p < 0.01) than δ15N of dogs; humans had lower δ13C values (p < 0.01) than dogs in Brasília, Piracicaba and Ubatuba. In Maraã, there was not any significant difference between dogs and humans. We concluded that CSA is still valid as a first approach in modern societies. However, isotopic differences found suggest that in modern societies processed dog food is increasingly disconnecting human and dog, jeopardising the use of CSA in the future if the trend of increasing processed dog food consumption continues to occur.

Carbon and nitrogen isotopic composition of commercial dog food in Brazil.

BACKGROUND: Brazil is a low- to medium-income country and has the second largest pet food market in the world with 8% of world pet food consumption. The lowest-income social class spends around 17% of their domestic budget on pet food and other items related to pets. Consumers are frequently misled by advertising as there is no precise information about the main sources of protein, carbohydrates and fat in the labels, and the Brazilian pet food industry can legally claim that their products contain certain items like salmon or beef even if they use just a flavoring compound. METHODS: The stable isotope methodology compares the stable isotope ratios of carbon (13C/12C) and nitrogen (15N/14N) between source and product. The 13C/12C ratio of a specific product (e.g., dog food) reveals the proportions of C4 (maize) and C3 (soybean, rice and wheat) plants in that product and the 15N/14N ratio reveals the proportion of the compounds derived from animals. With this isotopic data, we used MixSIAR, a Bayesian stable isotope-mixing model, to estimate the proportion of maize, grains, poultry and beef in dog food. RESULTS: The δ13C values of dry dog food ranged from -24.2‰ to -12.8‰, with an average (± standard-deviation) of -17.1‰ ± 2.8‰. The δ13C values of wet pet food ranged from -25.4‰ to -16.9‰, with an average (± standard-deviation) of -21.2‰ ± 2.4‰, which was significantly lower (p < 0.01). The δ15N values of the dry and wet food ranged from 1.7‰ to 4.2‰, and from 0.5‰ to 5.5‰, respectively. The average δ15N values of dry food (2.9‰ ± 0.5‰) was not higher than the wet food (2.6‰ ± 1.3‰) (p > 0.01). The output of the MixSIAR showed a low proportion of bovine products in dry dog food samples. On the other hand, poultry was obviously the dominant ingredient present in most of the samples. Maize was the second dominant ingredient. Wet and dry dog food showed similar isotopic analysis results. The only difference was a lower proportion of maize and higher proportion of grains in wet dog food. DISCUSSION: The main finding is that dog food in Brazil is mostly made of approximately 60% (ranging from 32% to 86%) animal-based and 40% (ranging from 14% to 67%) plant-based products. Poultry and maize are the main ingredients. Poultry is added as a by-product or meal, which avoids competition between dogs and humans for meat products, while they can compete for maize. On the other hand, a large proportion of plant-based products in dog food decreases the energy and environmental footprint, since plant-based food products tend to be less harmful compared to animal-based products. Labels can mislead consumers by showing pictures of items that are not necessarily part of the product composition and by not showing the detailed information on the proportion of each ingredient. This information would allow customers to make their own choices considering their pet's nutrition, the competition between animals and humans for resources and environmental sustainability.

Standardization of A Sample-Processing Methodology for Stable Isotope Studies in Poultry

The objective of this study was to determine if lipid extraction processes alter the isotopic value of 13C and 15N of tissues (pectoral muscle, thigh and liver) and eggs and if the use of anticoagulants interferes with blood and plasma 13C and 15N isotopic values. Samples were acquired from the same flock of birds. The 32 egg samples were randomly divided into four treatments (liquid, dehydrated, and fat-extracted with ether or chloroform + methanol) with eight replicates each. The 24 samples of pectoral muscle, thigh muscle and liver of broilers were randomly divided into three treatments (dehydrated, fat-extracted with ether and chloroform + methanol) with eight replicates each. Blood samples were divided into a 3x3 factorial arrangement with three physical forms (liquid, oven-dried or freeze-dried) and three collection methods (with no anticoagulant, with EDTA or heparin). Plasma samples were distributed in a 3x2 factorial arrangement, with three physical forms (liquid, oven-dried, or freeze-dried) and two anticoagulants (EDTA or heparin). The obtained isotopic results were submitted to the multivariate analysis of variance (MANOVA) and univariate (ANOVA, complemented by Tukey test), using the GLM procedure of the statistical program SAS (1996) or Minitab 16. The results show that it is possible to use the evaluated methods of fat extraction, drying and anticoagulants in the isotopic analyses of carbon-13 and nitrogen-15 in chicken tissues.

Standardization of A Sample-Processing Methodology for Stable Isotope Studies in Poultry

The objective of this study was to determine if lipid extraction processes alter the isotopic value of 13C and 15N of tissues (pectoral muscle, thigh and liver) and eggs and if the use of anticoagulants interferes with blood and plasma 13C and 15N isotopic values. Samples were acquired from the same flock of birds. The 32 egg samples were randomly divided into four treatments (liquid, dehydrated, and fat-extracted with ether or chloroform + methanol) with eight replicates each. The 24 samples of pectoral muscle, thigh muscle and liver of broilers were randomly divided into three treatments (dehydrated, fat-extracted with ether and chloroform + methanol) with eight replicates each. Blood samples were divided into a 3x3 factorial arrangement with three physical forms (liquid, oven-dried or freeze-dried) and three collection methods (with no anticoagulant, with EDTA or heparin). Plasma samples were distributed in a 3x2 factorial arrangement, with three physical forms (liquid, oven-dried, or freeze-dried) and two anticoagulants (EDTA or heparin). The obtained isotopic results were submitted to the multivariate analysis of variance (MANOVA) and univariate (ANOVA, complemented by Tukey test), using the GLM procedure of the statistical program SAS (1996) or Minitab 16. The results show that it is possible to use the evaluated methods of fat extraction, drying and anticoagulants in the isotopic analyses of carbon-13 and nitrogen-15 in chicken tissues.(AU)