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

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)

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.

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)

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.

Determinação de baixa inclusão de FCOB em dietas de poedeiras por isótopos estáveis / Assessment of low amounts of meat and bone meal in the diet of laying hens by using stable isotopes

The aim of this study was to assess whether the inclusion of low amounts of ingredients such as wheat bran, corn gluten and yeast in the diet of laying hens can interfere with the traceability of meat and bone meal (MBM) in eggs and its fractions (albumen and yolk). We used 256 laying hens distributed across eight treatment groups, which consisted of a diet based on corn-soybean-wheat (CSW) bran meal and other diets that had additions comprising gluten and/or (MBM) and/or yeast. To analyse the isotopic ratios (13C:12C and 15N:14N), on the 28th and 56th days, 16 eggs were randomly taken for each treatment (four for each repetition), where eight eggs (two for each repetition) were used to harvest yolk and albumen samples and the remaining eight (two for each repetition) were used to analyse the whole egg. We concluded that detection of 2.0% MBM in the egg and its fractions is possible; however, the results for treatments containing gluten and yeast were not different from those containing MBM. Therefore, the inclusion of 3.0% gluten and/or yeast does not result in different ?13C and ?15N isotopic values in the egg and its fractions compared to the values obtained with the addition of 2.0% MBM in the diet.(AU)

O objetivo foi verificar se a baixa inclusão de outros ingredientes, como farelo de trigo, glúten de milho e levedura na dieta de poedeiras, pode interferir na rastreabilidade da farinha de carne e ossos bovinos (FCOB) em ovos e suas frações (albúmen e gema). Foram utilizadas 256 galinhas poedeiras distribuídas em oito tratamentos, no qual, foram compostos de uma dieta MST a base de milho, farelo de soja e farelo de trigo e as demais dietas foram acrescidos de glúten e/ou (FCOB) e/ ou levedura. Para análise da razão isotópica (13C/12C e 15N/14N), nos 28º e 56º dias foram tomados aleatoriamente 16 ovos por tratamento (quatro por repetição), onde oito ovos (dois por repetição) serviram para colheitas de amostras de gema e albúmen e os outros oito (dois por repetição) para ovo total. Pode-se concluir que, a detecção de 2,0% de FCOB no ovo e suas frações é possível, porém, os tratamentos com inclusão de glúten e levedura não foram diferentes dos tratamentos com inclusão da FCOB. Portanto, a inclusão de 3,0% de glúten e ou levedura na ração, não permite a diferenciação dos valores isotópicos de ?13C e ?15N do ovo e suas frações destes grupos dos que possuem 2,0% FCOB.(AU)

The use of carbon and nitrogen stable isotopes for the detection of poultry offal meal in meat-type quail feeds

The objective of the present study was to trace the inclusion of poultry offal meal (POM) in the diet of meat-type quails reared for a long period using the technique of stable isotopes. A number of 320 quails were randomly distributed into eight treatments: vegetable diet (T1), and a diet containing 8% POM were fed until the end of the experimental period (T2) or replaced by the vegetable diet on day 42 (T3), 56 (T4), 70 (T5), 84 (T6), 98 (T7), and 112 (T8). Breast muscle samples were collected from four birds randomly selected per treatment every 14 days. The obtained isotope results were submitted to multivariate analysis of variance (MANOVA) with the aid of the GLM procedure of statistical SAS program. Treatments were different from T1 when birds were sacrificed at least two weeks after the diet was changed. T2 results were different from T1 in all evaluated periods. It was concluded that it is possible to trace poultry offal meal inclusion in a strictly vegetable diet after the diet was changed for at least 14 days.

The use of carbon and nitrogen stable isotopes for the detection of poultry offal meal in meat-type quail feeds

The objective of the present study was to trace the inclusion of poultry offal meal (POM) in the diet of meat-type quails reared for a long period using the technique of stable isotopes. A number of 320 quails were randomly distributed into eight treatments: vegetable diet (T1), and a diet containing 8% POM were fed until the end of the experimental period (T2) or replaced by the vegetable diet on day 42 (T3), 56 (T4), 70 (T5), 84 (T6), 98 (T7), and 112 (T8). Breast muscle samples were collected from four birds randomly selected per treatment every 14 days. The obtained isotope results were submitted to multivariate analysis of variance (MANOVA) with the aid of the GLM procedure of statistical SAS program. Treatments were different from T1 when birds were sacrificed at least two weeks after the diet was changed. T2 results were different from T1 in all evaluated periods. It was concluded that it is possible to trace poultry offal meal inclusion in a strictly vegetable diet after the diet was changed for at least 14 days.(AU)

The use of carbon and nitrogen stable isotopes for the detection of poultry offal meal in meat-type quail feeds

The objective of the present study was to trace the inclusion of poultry offal meal (POM) in the diet of meat-type quails reared for a long period using the technique of stable isotopes. A number of 320 quails were randomly distributed into eight treatments: vegetable diet (T1), and a diet containing 8% POM were fed until the end of the experimental period (T2) or replaced by the vegetable diet on day 42 (T3), 56 (T4), 70 (T5), 84 (T6), 98 (T7), and 112 (T8). Breast muscle samples were collected from four birds randomly selected per treatment every 14 days. The obtained isotope results were submitted to multivariate analysis of variance (MANOVA) with the aid of the GLM procedure of statistical SAS program. Treatments were different from T1 when birds were sacrificed at least two weeks after the diet was changed. T2 results were different from T1 in all evaluated periods. It was concluded that it is possible to trace poultry offal meal inclusion in a strictly vegetable diet after the diet was changed for at least 14 days.

Traceability of animal meals in Japanese quail eggs using the technique of 13C e 15N* stable isotopes

The objective of this study was to trace the inclusion of bovine meat and bone meal (BMBM) in the diet of Japanese quails by analyzing eggs and egg fractions (yolk and albumen) by the technique of carbon-13 (13C) and nitrogen-15 (15N) stable isotopes. In the trial, 120 Japanese quails were distributed in six treatments with four replicates of five birds each. The following treatments were applied: feed based on corn and soybean meal, containing graded BMBM inclusions (0, 1, 2, 3, 4 or 5%). After 42 days, 20 eggs per treatment were randomly collected for three consecutive days. Ten eggs were used for yolk and albumen sample collection, and ten for total egg sample collection. It was possible to detect the dietary inclusion of 1% BMBM in the egg and its fractions. Therefore, the technique of isotopes 13C and 15N is able of tracing since 1% inclusion level of BMBM in the diet of Japanese quails in eggs and their fractions.

Traceability of animal meals in Japanese quail eggs using the technique of 13C e 15N* stable isotopes

The objective of this study was to trace the inclusion of bovine meat and bone meal (BMBM) in the diet of Japanese quails by analyzing eggs and egg fractions (yolk and albumen) by the technique of carbon-13 (13C) and nitrogen-15 (15N) stable isotopes. In the trial, 120 Japanese quails were distributed in six treatments with four replicates of five birds each. The following treatments were applied: feed based on corn and soybean meal, containing graded BMBM inclusions (0, 1, 2, 3, 4 or 5%). After 42 days, 20 eggs per treatment were randomly collected for three consecutive days. Ten eggs were used for yolk and albumen sample collection, and ten for total egg sample collection. It was possible to detect the dietary inclusion of 1% BMBM in the egg and its fractions. Therefore, the technique of isotopes 13C and 15N is able of tracing since 1% inclusion level of BMBM in the diet of Japanese quails in eggs and their fractions.(AU)

Traceability of animal meals in Japanese quail eggs using the technique of 13C e 15N* stable isotopes

The objective of this study was to trace the inclusion of bovine meat and bone meal (BMBM) in the diet of Japanese quails by analyzing eggs and egg fractions (yolk and albumen) by the technique of carbon-13 (13C) and nitrogen-15 (15N) stable isotopes. In the trial, 120 Japanese quails were distributed in six treatments with four replicates of five birds each. The following treatments were applied: feed based on corn and soybean meal, containing graded BMBM inclusions (0, 1, 2, 3, 4 or 5%). After 42 days, 20 eggs per treatment were randomly collected for three consecutive days. Ten eggs were used for yolk and albumen sample collection, and ten for total egg sample collection. It was possible to detect the dietary inclusion of 1% BMBM in the egg and its fractions. Therefore, the technique of isotopes 13C and 15N is able of tracing since 1% inclusion level of BMBM in the diet of Japanese quails in eggs and their fractions.

TURNOVER ISOTÓPICO EM LEITE, SANGUE E FEZES DE VACAS LEITEIRAS ALIMENTADAS COM CAMA DE AVIÁRIO

Resumo O objetivo este estudo foi avaliar o turnover isotópico de 13C e 15N no leite, sangue e fezes, ocasionados pela inclusão da cama de aviário na dieta de vacas leiteiras, a fim de determinar o tempo de incorporação do valor isotópico da dieta. Foram utilizadas 12 vacas mestiças, distribuídas em delineamento inteiramente casualizado com seis repetições e dois tratamentos. Os tratamentos foram: T1 dieta estritamente vegetal durante todo o período experimental; T2 50% cama de aviário e 50% vegetal durante o primeiro período experimental e depois dieta vegetal. Foram colhidas amostras diariamente e analisadas com espectrômetro de massas de razão isotópica. Para mensurar o turnover de 13C e 15N em determinado intervalo de tempo, foi utilizada a função exponencial de primeira ordem. Os dados isotópicos foram submetidos ao teste t-Student. Os valores isotópicos das fezes e do leite atingiram o patamar de equilíbrio. O sangue não conseguiu atingir o patamar de equilíbrio nos períodos avaliados indicando ser um tecido para detecção de dietas de períodos mais longos. O resultado do teste de t Student (p<0,05) foi semelhante ao encontrado pela análise dos isótopos estáveis mostrando que no primeiro período os valores isotópicos das fezes do T1 diferiram do T2, e para o leite não houve diferença entre os tratamentos. No segundo período, os valores isotópicos das fezes e do leite do T1 não diferiram do T2. Os valores isotópicos de 15N no leite para o 1°período diferiram entre o T1 e o T2. De acordo com os resultados obtidos pode-se concluir que o turnover isotópico do 13C nas fezes e do 15N no leite indicam valores de meia vida de 1,4 dias quando houve a troca da dieta contendo cama de aviário para a somente vegetal.

The objective of this study was to evaluate the isotopic turnover of 13C and 15N in milk, blood and feces, caused by the inclusion of poultry litter in the diet of dairy cows, in order to determine the incorporation time of the isotopic value of the diet. Were used twelve crossbred cows, distributed in a completely randomized design with six replicates and two treatments. The treatments were T1 - strictly vegetable diet throughout the experimental period; T2 50% poultry litter an 50% vegetable during the first experimental period and after vegetable diet. Samples were collected daily and analyzed with isotope ratio mass spectrometers. The first order exponential function was used to measure the turnover of 13C and 15N in a certain period of time. Were submitted the isotopic data to Student's t-test. The isotopic values of feces and milk reached the plateau isotope. The blood was not able to reach the plateau isotope in the evaluated periods indicating to be a tissue for detection of diets of longer periods. The results of the Students t-test (p <0.05) were similar to those found by the stable isotope analysis, showing that in the first period, the isotopic values of the feces of the T1 differed from the T2, and for the milk there was no difference between the treatments. In the second period, the isotopic values of the feces and the milk of the T1 did not differ from the T2. The isotopic values of 15N in milk for the 1st period differed between T1 and T2. According to the results obtained, it can be concluded that the isotopic turnover of 13C in feces and 15N in milk indicate half-life values of 1.4 days when there was a change from diet containing poultry litter to vegetable only.

Multivariate analysis of stable isotope data in the traceability process for birds - doi: 10.4025/actascianimsci.v34i4.13434

Isotope analysis has proved to be an extremely important tool in the traceability process; however, statistical analyses of the results show discrepancies, as the data depend on and originate from several chemical elements such as carbon, hydrogen, oxygen, nitrogen and sulfur (CHONS). In order to establish the proper analysis of traceability data for birds using the stable isotope technique and evaluate the need for a combined analysis of the variables, data for carbon-13 and nitrogen-15 were used from eggs (albumen + yolk) of laying hens and the pectoral muscle of broilers, which were subjected to univariate statistical analysis (ANOVA and complemented with Tukeys test) and multivariate statistical analysis (MANOVA and Discriminant Analysis). The data were analyzed using Minitab 16 software, and the results, corroborated in the theory, confirm the need for multivariate analysis, showing also that discriminant analysis clarifies questions from the results of the other analysis methods compared in this study.

Multivariate analysis of stable isotope data in the traceability process for birds / Análise multivariada em dados de isótopos estáveis no processo de rastreabilidade em aves

Isotope analysis has proved to be an extremely important tool in the traceability process; however, statistical analyses of the results show discrepancies, as the data depend on and originate from several chemical elements such as carbon, hydrogen, oxygen, nitrogen and sulfur (CHONS). In order to establish the proper analysis of traceability data for birds using the stable isotope technique and evaluate the need for a combined analysis of the variables, data for carbon-13 and nitrogen-15 were used from eggs (albumen + yolk) of laying hens and the pectoral muscle of broilers, which were subjected to univariate statistical analysis (ANOVA and complemented with Tukey's test) and multivariate statistical analysis (MANOVA and Discriminant Analysis). The data were analyzed using Minitab 16 software, and the results, corroborated in the theory, confirm the need for multivariate analysis, showing also that discriminant analysis clarifies questions from the results of the other analysis methods compared in this study.

A análise isotópica tem se mostrado uma ferramenta de suma importância ao processo de rastreabilidade, no entanto, existem divergências nas análises estatísticas dos resultados, uma vez que os dados são dependentes e advindos de vários elementos químicos tais como Carbono, Hidrogênio, Oxigênio, Nitrogênio e Enxofre (CHON'S). Com o intuito de estabelecer a análise propícia para os dados de rastreabilidade em aves pela técnica de isótopos estáveis e avaliar a necessidade da análise conjunta das variáveis, foram usados dados de carbono-13 e de nitrogênio-15 de ovos (albúmen + gema) de poedeiras e músculo peitoral de frangos de corte, os quais foram submetidos à análise estatística univariada (Anova e complementada pelo teste de Tukey) e multivariada (Manova e Discriminante). Os dados foram analisados no software Minitab 16, e os resultados, consolidados na teoria, confirmam a necessidade de análise multivariada, mostrando também que a análise discriminante esclarece as dúvidas apresentadas nos resultados de outros métodos de análise comparados nesta pesquisa.

Isotopic traceability in body tissues and in different breeding periods of broilers / Rastreabilidade isotópica em tecidos corporais e em diferentes períodos de criação de frangos de corte

The inclusion of poultry viscera meal (VM) in broiler diets to possibly replace it with strictly vegetable diet (VE) and vice-versa, using the technique of carbon and nitrogen stable isotopes in different tissues was traced. One hundred ninety-two one-day-old chicks that had been distributed randomly were used in twelve experimental treatments with four replications of four chicks each. The treatments consisted of vegetable diet (VG) for diets containing FV going after a certain age or the reverse, in which the birds began feeding VM on diet and then switched to VE diet. At 42 days of age, samples of pectoral muscle (Pectoralis major), keel and tibia were collected, with later determination of isotopic ratios (13C/12C and 15N/14N). The stable isotopes technique couldn't trace the use of VM in feeding broilers when this ingredient was part of broiler diets only in the first seven, 14 or 21 days of age. However, this technique can be applied to younger poultry that have been sampled before a possible change in diet, since they can have the isotopic signature of feeding stabilized around two weeks of age.(AU)

Rastrearam-se a inclusão de farinha de vísceras de aves (FV) em dietas de frangos por ocasião de eventual substituição de dieta contendo FV por dieta estritamente vegetal, e vice-versa, por isótopos estáveis de carbono e de nitrogênio. Foram distribuídos aleatoriamente 192 pintos de um dia de idade, em 12 tratamentos com quatro repetições de quatro aves. Os tratamentos constituíam-se de dieta de vegetais (VG) passando para dietas contendo FV, após certa idade, ou o inverso, em que as aves começaram se alimentando de dieta FV e depois passaram para dieta VG. Aos 42 dias de idade, foram coletadas amostras de músculo peitoral (Pectoralis major), quilha e tíbia, para determinação das razões isotópicas (13C/12C e 15N/14N). A técnica dos isótopos estáveis somente não foi capaz de rastrear a utilização de FV na alimentação de frangos de corte, quando esse ingrediente fez parte da dieta das aves apenas nos primeiros sete, 14 ou 21 dias de idade. Entretanto, há a possibilidade da aplicação dessa técnica em aves mais jovens, amostradas antes de eventual mudança de dieta, pois elas podem ter a assinatura isotópica da alimentação estabilizada em torno de duas semanas de idade.(AU)

FRANGOS DE CORTE DE PRODUTORES DO ESTADO DE SÃO PAULO: RASTREABILIDADE DA ALIMENTAÇÃO POR DELTA13C e DELTA15N

RASTREABILIDADE DE SUBPRODUTOS DE ORIGEM ANIMAL NO MÚSCULO PEITORAL E MUCOSA INTESTINAL DE FRANGOS DE CORTE DE GRANJAS COMERCIAIS POR 13Ce 15N Objetivou-se rastrear a presença de subprodutos de origem animal na alimentação de frangos de corte de granjas comerciais por meio da metodologia dos isótopos estáveis de carbono e nitrogênio (13Ce 15N), no músculo peitoral e mucosa intestinal, assim como iniciar um banco de dados de valores bromatológicos e isotópicos dos ingredientes da ração de maior interesse na avicultura de corte. Foram amostradas aleatoriamente oito aves (machos) semanalmente de sete granjas de diferentes empresas avícolas dentro do Estado de São Paulo para obtenção das amostras de músculo peitoral e mucosa intestinal. Nos mesmos dias de coletas foram retiradas três amostras aleatórias de ração, adquiridas dos comedouros de cada granja. Para o banco de dados foram amostrados os ingredientes milho, soja, farelo de soja e farinhas de origem animal dos departamentos de recepção e controle de qualidade dos ingredientes das fabricas de ração visitadas. Todas as amostras foram submetidas à análise de espectrometria de massa de razão isotópica, sendo que a ração também foi avaliada quanto à presença de DNA de origem animal por PCR. Os dados isotópicos das amostras foram submetidos à análise de variância em cada idade de coleta, sendo complementados pelo teste de Tukey, e o conjunto de todos os dados foram avaliados pelas análises multivariadas de componentes principais e discriminante linear. Foi possível verificar pela análise de PCR na ração que somente as empresas D e F utilizaram ração estritamente vegetal ao longo do período de criação das aves. Houve diferença entre os valores isotópicos da ração, músculo peitoral e mucosa intestinal das aves das empresas D e F em relação às demais, sendo esta mais definida para o 15N dos sete aos 42 dias de idade. Esse mesmo resultado foi obtido pelas análises multivariadas. Portanto, foi possível diferenciar as empresas que utilizaram alimentação estritamente vegetal em relação às demais, demonstrando a possibilidade de rastrear a presença de subprodutos de origem animal na alimentação dos frangos de corte de granjas comerciais pela metodologia dos isótopos estáveis. DETECÇÃO DE SUBPRODUTOS DE ORIGEM ANIMAL POR 13C e 15N NO SANGUE E PENAS DE FRANGOS DE CORTE DE GRANJAS COMERCIAIS A metodologia dos isótopos estáveis tem-se mostrado uma ferramenta analítica promissora na autentificação de alimentos e poderia auxiliar a testar a veracidade das informações constantes no histórico alimentar de lotes de produtos diferenciados, como os frangos com alimentação estritamente vegetal. Entretanto, há necessidade de estudos de campo com animais de granjas comerciais assim como amostragem de tecidos/frações corporais sem necessidade de sacrifícios dos animais. Portanto, objetivou-se rastrear a presença de subprodutos de origem animal na alimentação de frangos de corte de granjas comerciais, por meio da metodologia dos isótopos estáveis de carbono e nitrogênio (13C e 15N) no sangue e penas. Foram amostradas aleatoriamente oito aves (machos) semanalmente de sete granjas de diferentes empresas avícolas dentro do Estado de São Paulo para obtenção das amostras de sangue e penas. Nos mesmos dias de coletas foram retiradas três amostras aleatórias de ração, adquiridas dos comedouros de cada granja. Todas as amostras foram submetidas à análise de espectrometria de massa de razão isotópica, sendo que a ração também foi avaliada quanto à presença de DNA de origem animal por PCR. Os dados isotópicos das amostras foram submetidos à análise de variância em cada idade de coleta, e o conjunto de todos os dados foram avaliados pelas análises multivariadas de componentes principais e discriminante linear. Foi possível verificar pela análise de PCR na ração que somente as empresas D e F utilizaram ração estritamente vegetal ao longo do período de criação das aves e que houve diferença significativa entre os valores isotópicos da ração, sangue e penas das aves das empresas D e F em relação às demais pela análise de variância para o 15N e pelas análises multivariadas testadas dos sete aos 42 dias de idade. Portanto, foi possível diferenciar as empresas que utilizaram alimentação estritamente vegetal em relação às demais, demonstrando a possibilidade de rastrear a presença de subprodutos de origem animal na alimentação dos frangos de corte de granjas comerciais pela metodologia dos isótopos estáveis.

TRACEABILITY OF ANIMAL BY-PRODUCTS IN PECTORAL MUSCLE AND INTESTINAL MUCOSA OF COMMERCIAL BROILER FARMS BY 13CAND 15N The aim of this study was to trace the presence of animal by-products in commercial broiler farms feed using the methodology of stable isotope ratios of carbon and nitrogen (13Cand 15N) in the pectoral muscle and intestinal mucosa, and start a database of bromatological and isotopic values of feed ingredients with greatest interest in poultry production. Eight male birds were sampled randomly week-by-week in seven broiler farms of different poultry companies in the São Paulo State to obtain samples of pectoral muscle and intestinal mucosa. Were taken on the same day of sampling three random samples of feed, acquired from feeders of each commercial broiler unit. For the database were sampled corn, soybean, soybean meal and animal by-products obtained from reception and quality control departments of visited feed factories. All samples were analyzed using isotope ratio mass spectrometry, and feed was also evaluated for the presence of animal DNA by PCR analysis. The isotopic data of the samples were submitted to analysis of variance in each age collection, complemented by Tukey test, and the set of all data were analyzed by multivariate analysis of major and linear discriminant components. It was verified by PCR analysis in feed that only D and F companies used strictly vegetable diet throughout the period of raising birds. There was difference between isotopic values of feed, pectoral muscle, and intestinal mucosa of birds from D and F companies compared with others, which is most definitely for the 15N from seven to 42 days old. The same result was obtained by multivariate analysis. Therefore, it was possible to differentiate the companies that used strictly vegetable diet from the others, demonstrating the ability to trace the presence of animal by-products in commercial broiler farms feed using the methodology of stable isotopes. Keywords: multivariate analysis, PCR, stable isotope DETECTION OF ANIMAL BY-PRODUCTS BY 13CAND 15N IN BLOOD AND FEATHERS OF COMMERCIAL BROILER FARMS The methodology of stable isotopes has been shown to be a promising analytical tool in authentication of food and could help test the veracity of the information contained in the food history of lots of different products such as chickens strictly vegetable diet. However, there is need for animal studies field of commercial farms as well as sampling tissue/body fractions without sacrifice of the animals. Therefore, the goal of this study was to investigate the presence of animal by-products in commercial broiler farms feed, using the methodology of stable isotope ratios of carbon and nitrogen (13Cand 15N) in blood and feathers. Eight male birds were sampled randomly week-by-week in seven broiler farms of different poultry companies in the São Paulo State to obtain samples of blood and feathers. Were taken on the same day of sampling three random samples of feed, acquired from feeders of each commercial broiler unit. All samples were analyzed using isotope ratio mass spectrometry, and feed was also evaluated for the presence of animal DNA by PCR analysis. The isotopic data of the samples were submitted to analysis of variance in each age collection, and the set of all data were analyzed by multivariate analysis of major and linear discriminant components. It was verified by PCR analysis in feed that only D and F companies used strictly vegetable diet throughout the period of raising birds and that was a significant difference between isotopic values of feed, blood, and feathers of birds from D and F companies compared with others by 15N analysis of variance and multivariate analysis tested from seven to 42 days old. Therefore, it was possible to differentiate the companies that used strictly vegetable diet from the others, demonstrating the ability to trace the presence of animal by-products in commercial broiler farms feed using the methodology of stable isotopes. Keywords: analysis of main components, carbon-13, nitrogen-15

Traceability of poultry offal meal in broiler feeding using isotopic analysis (13C and 15N) of different tissues

Our goal was to trace the inclusion of poultry offal meal (OM) in diets by using carbon (13C/12C) and nitrogen (15N/14N) isotopic ratios of different tissues in order to contribute for the development of an independent technology for the certification of the feeding of broilers reared on diets with no addition of animal ingredients. Eighty one-day-old chicks were randomly distributed into five experimental treatments, that is, diets containing increasing levels of OM inclusion (0, 2, 4, 8 and 16% OM), with four replicates of four birds each. At 42 days of age, four birds per treatment (n=4) were randomly selected, weighed, and sacrificed to collect breast muscle (Pectoralis major), keel and tibia samples to determine their isotopic ratios (13C/12C e 15N/14N). It was observed that 13C and 15N enrichment increased as a function of increasing OM inclusion in all diets. The analyses of the Pectoralis major showed that that only treatments with 8 and 16% OM dietary inclusion were different form those in the control group (0% OM). On the other hand, when the keel and tibia were analyzed, in addition to 8 and 16% OM), the treatment with 4% OM inclusion was also different from the control group. The use of isotopic ratios of stable carbon and nitrogen isotopes is an alternative to trace OM inclusion in broiler diets as it is capable of tracing OM levels below those usually practiced by the poultry industry in Brazil.

Traceability of poultry offal meal in broiler feeding using isotopic analysis (13C and 15N) of different tissues

Our goal was to trace the inclusion of poultry offal meal (OM) in diets by using carbon (13C/12C) and nitrogen (15N/14N) isotopic ratios of different tissues in order to contribute for the development of an independent technology for the certification of the feeding of broilers reared on diets with no addition of animal ingredients. Eighty one-day-old chicks were randomly distributed into five experimental treatments, that is, diets containing increasing levels of OM inclusion (0, 2, 4, 8 and 16% OM), with four replicates of four birds each. At 42 days of age, four birds per treatment (n=4) were randomly selected, weighed, and sacrificed to collect breast muscle (Pectoralis major), keel and tibia samples to determine their isotopic ratios (13C/12C e 15N/14N). It was observed that 13C and 15N enrichment increased as a function of increasing OM inclusion in all diets. The analyses of the Pectoralis major showed that that only treatments with 8 and 16% OM dietary inclusion were different form those in the control group (0% OM). On the other hand, when the keel and tibia were analyzed, in addition to 8 and 16% OM), the treatment with 4% OM inclusion was also different from the control group. The use of isotopic ratios of stable carbon and nitrogen isotopes is an alternative to trace OM inclusion in broiler diets as it is capable of tracing OM levels below those usually practiced by the poultry industry in Brazil.

Traceability of bovine meat and bone meal in poultry by stable isotope analysis

Bovine meat and bone meal (MBM) was widely used in animal diets until outbreaks of Bovine Spongiform Encefalopathy (BSE) occurred in some countries. It has not been confirmed yet whether or not BSE may be transmitted to man through chicken meat originated from poultry that had been fed diets containing MBM. Therefore, consumers nowadays express preference for meat originated from birds fed exclusively vegetable diets. This study analyzed samples of major breast muscle (Pectoralis major) using mass spectrometry of stable isotopes (carbon and nitrogen) as a means to assess the presence of MBM in broiler diets, a technique that might be used in the certification of poultry quality. A total of 150 day-old chicks were reared in five randomized treatments with increasing MBM dietary inclusion levels (0, 1, 2, 4 and 8%). On day 42, breast muscle samples were collected from three birds per treatment and used in the determination of 13C/12C and 15N/14N isotope ratios. The breast muscle isotope values were expressed as delta in parts per thousand (delta‰). The following carbon isotope values (13C) were found: 18.74‰±0.11, 18.51‰±0.19, 18.24‰±0.10, -17.79‰ ±0.12, and -17.15‰±0.15 for 0, 1, 2, 4 and 8% MBM dietary levels, respectively. Nitrogen isotope values (15N) were 1.65‰±0.14, 1.65‰±0.28, 1.72‰±0.08, 1.95‰±0.16, and 2.52‰ ± 0.09 for 0, 1, 2, 4 and 8% MBM dietary levels, respectively. This study showed important differences in delta13C and delta15N values in breast meat, evidencing a simultaneous enrichment of this isotopic pair, which allowed tracing MBM in bird diets. Analysis of carbon and nitrogen stable isotopes may be used to ensure feeding with exclusively vegetable diets, and might also be used as a reliable evaluation tool in broiler meat certification. The diet with 1% inclusion level of MBM and the exclusively vegetable diet showed similar results.