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Secreção de hormônio de crescimento de camundongo por queratinócitos humanos primários: perspectivas para um modelo animal de terapia gênica cutânea / Secretion of mouse growth hormone by transduced primary human keratinocytes: prospects for an animal model of cutaneous gene therapyCecchi, Claudia Regina 05 September 2008 (has links)
Queratinócitos são um veículo bastante atrativo para a transferência gênica ex vivo e liberação sistêmica uma vez que as proteínas secretadas por estas células podem atingir a circulação via um mecanismo similar ao processo natural. Um eficiente vetor retroviral (LXSN) contendo o gene do hormônio de crescimento de camundongo (mGH) foi utilizado para transduzir queratinócitos humanos primários. Os queratinócitos transduzidos apresentaram um nível de secreção in vitro alto e estável atingindo até 11 g mGH/106 células/dia. Os epitélios formados por estes queratinócitos geneticamente modificados apresentaram, porém, uma queda na taxa de secreção > 80 % quando foram retirados da placa de cultura utilizando um procedimento clássico. A substituição desta metodologia clássica por uma cultura organotípica resolveu completamente este problema. Camundongos anões imunodeficientes (lit/scid) implantados com estes enxertos organotípicos foram acompanhados durante 4 meses, e apresentaram um aumento de peso significativo (P<0,05) nos primeiros 40 dias. Níveis circulatórios de mGH atingiram um pico de 21 ng/mL 1 h após o implante, mas estes níveis rapidamente atingiram níveis basais (~2 ng/mL). Os queratinócitos humanos primários apresentaram portanto altos níveis de expressão in vitro e os maiores níveis circulatórios, porém por um breve período de tempo, reportados até o momento para GH neste tipo de células. Em conjunto com resultados que mostraram uma recuperação considerável da eficiência de secreção de mGH em cultura por enxertos organotípicos retirados dos animais, foram discutidos os fatores que ainda impedem a utilização clínica deste modelo promissor de terapia gênica cutânea. / Keratinocytes are a very attractive vehicle for ex vivo gene transfer and systemic delivery, since proteins secreted by these cells may reach the circulation via a mechanism which mimics the natural process. An efficient retroviral vector (LXSN) encoding the mouse growth hormone gene (mGH) was used to transduce primary human keratinocytes. Organotypic raft cultures were prepared with these genetically modified keratinocytes and were grafted onto immunodeficient dwarf mice (lit/scid). Transduced keratinocytes presented a high and stable in vitro secretion level of up to 11 g mGH/106cells/day. Conventional epidermal sheets made with these genetically modified keratinocytes, however, showed a drop in secretion rates of > 80% simply due to detachment of the epithelium from its substratum. Substitution of conventional grafting methodologies with organotypic raft cultures completely overcame this problem. The stable long-term grafting of such cultures onto immunodeficient dwarf (lit/scid) mice could be followed for more than 4 months, and a significant weight increase (P<0.05) over the control group was observed in the first 40 days. Circulating mGH levels revealed a peak of 21 ng/mL just 1h after grafting, but unfortunately these levels rapidly fell to baseline values (~ 2 ng/mL). mGH-secreting primary human keratinocytes presented the highest in vitro expression and peak circulatory levels reported to date for a form of GH with this type of cells. Together with data showing that excised implants can recover in culture a remarkable fraction of their original in vitro mGH secretion efficiency, the factors that might still hamper the success of this promising model of cutaneous gene therapy are discussed. SUMÁRIO
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Associação entre polimorfismos genéticos e parâmetros da curva de crescimento em bovinos de corte. / Association between genetic polymorphisms and growth curve parameters in beef cattle.Claudia Cristina Paro de Paz 13 December 2002 (has links)
Foram analisadas informações provenientes de um experimento de avaliação de sistemas cruzamento entre raças bovinas de corte, executado na Embrapa Pecuária Sudeste, com o objetivo de avaliar o efeito dos polimorfismos genéticos da kappa-caseína-HinfI (CSN3): AA e AB, do hormônio do crescimento-AluI (GH): LL e LV e da b-lactoglobulina-HaeIII (LGB): AA, AB e BB sobre a curva de crescimento de bovinos de três grupos genéticos, ½Nelore+½Canchim (NC), ½Nelore+½Aberdeen-Angus (NA) e ½Nelore+½Simental (NS), nascidos nos anos de 1998 e 1999. Os pesos foram medidos ao nascimento, ao desmame e mensalmente dos 8 aos 19 meses de idade. As análises foram realizadas por meio de duas abordagens. Na primeira, as estimativas dos parâmetros A (valor assintótico), k (taxa de maturação) e m (ponto de inflexão) obtidas do modelo Logístico, ajustado para descrever o crescimento de cada animal, foram analisados usando um modelo linear, que incluiu além do efeito do genótipo, o qual foi formado pela concatenação dos polimorfismos genéticos de CSN3, GH e LGB (G1=AALLAA, G2=AALLAB, G3=AALLBB, G4=AALVAB, G5=AALVBB, G6=ABLLAA, G7=ABLLAB, G8=ABLLBB, G9=ABLVAA, G10=ABLVAB e G11=ABLVBB), o ano de nascimento, o sexo e o manejo alimentar. Para os animais do grupo genético NC, os genótipos influenciaram significativamente as estimativas dos parâmetros A e k da curva de crescimento. O genótipo G3 apresentou valor inferior de A e superior de k em relação aos genótipos G7 e G8. Na segunda abordagem, os dados foram analisados por modelos não lineares, incluindo no modelo, os efeitos fixos de grupo contemporâneo e de genótipo dos genes CSN3 GH e LGB, com o objetivo de verificar a influência destes genes sobre a curva de crescimento destes animais e estimar os parâmetros da função Logística, simultaneamente. Os resultados deste estudo sugerem que a função Logística utilizada para descrever o crescimento dos grupos genéticos NC, NA e NS, foi influenciada pelos genótipos dos genes CSN3, GH e LGB. As maiores diferenças entre os genótipos dos genes CSN3, GH e LGB foram encontradas a partir dos 12-13 meses de idade. O genótipo AA para CSN3 apresentou maior taxa de maturação (k) que o genótipo AB nos grupos genéticos NC, NA e NS. Quanto ao valor assintótico (A), diferença entre AA e AB, foi pequena nos grupos genéticos NC e NS. Para o polimorfismo do GH no grupo genético NA, a curva que descreve o crescimento dos animais do genótipo LL mostrou-se mais desejável, do ponto de vista de produção de bovinos de corte. Entretanto, ocorreu uma inversão desta tendência no grupo genético NS. O mesmo ocorreu para o LGB, em que os genótipos AA e AB apresentaram estimativas do parâmetro k superiores em relação ao genótipo BB no grupo genético NA, enquanto o genótipo AB apresentou estimativa de k inferior em relação ao genótipo BB, no grupo genético NS. Evidências de que os polimorfismos dos genes CSN3, GH e LGB estejam ligado a QTL influenciando a curva de crescimento de bovinos, indicam que no futuro os genótipos destes genes podem ser usados em programas de seleção assistida por marcadores. / Data from a crossbreeding experiment, carried out at Embrapa Southeast Cattle Research Center, State of São Paulo, Brazil, were analyzed in order to evaluate the effects of kappa-casein-HinfI (CSN3): AA and AB, growth hormone-AluI (GH): LL and LV, and b-lactoglobulin-HaeIII (LGB): AA, AB and BB, polymorphism on the growth curve of three beef cattle crosses, ½Nellore + ½Canchim (NC), ½Nellore + ½Aberdeen Angus (NA) and ½Nellore + ½Simmental (NS). Animals were born in the years 1998 and 1999. Weight measurements were collected at birth, weaning (7 months of age) and monthly from 8 to 19 months of age. The effect these genetic polymorphisms were analyzed by two approaches. In the first one, the parameters A (asymptotic value), k (maturing rate) and m (inflection point) estimated by Logistic model for each animal, were analyzed by least squares, fitting a linear model which included the fixed effects of the genotype, which was identified by combination of polymorphic RFLPs of the genes CSN3, GH e LGB (G1=AALLAA, G2=AALLAB, G3=AALLBB, G4=AALVAB, G5=AALVBB, G6=ABLLAA, G7=ABLLAB, G8=ABLLBB, G9=ABLVAA, G10=ABLVAB e G11=ABLVBB), year of birth, sex and feed management. For the NC genetic groups, was detected significant effect of genotypes groups for A and k parameters estimates. The genotypes G3 presented inferior value of A and superior of k in relation to G7 and G8. In the second approach, it was fitted a Logistic non linear model which included the fixed effects of the contemporary group and genotype of the genes CSN3, GH and LGB, to examine the effect of these markers on growth curve parameters and to obtain the parameters estimates of the Logistic function, simultaneously. The growth curve used to explain the growth of the NC, NA and NS genetic groups, was influenced by genotypes of the CSN3, GH e LGB markers The major differences started at 12-13 months of age. The value of the maturing rate (k) of the AA genotype for CSN3 was superior in relation to AB genotype in the NC, NA and NS genetic groups. However, there was observed small difference in estimate of the asymptotic value (A) for the AA and AB genotypes in NC and NS genetic groups. For the GH polymorphism in NA genetic group the growth curve of the LL genotype showed more desirable to production of the beef cattle. The same was observed for the LGB, there were superior values of the parameter k of the AA and AB genotypes in relation to the BB for the NA genetic group, however there was inferior value of the parameter k of the AB genotype in relation to the BB, for the NS genetic group. Evidences that CSN3, GH and LGB polymorphism are linked to QTL influencing growth curve in beef cattle, indicates these genotypes may be a useful marker in future maker-assisted selection programs.
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Estudo do desempenho animal e produção do capim Tanzânia (Panicum maximum, Jacq.) em um sistema rotacionado de pastejo sob irrigação em três níveis de resíduo pós-pastejo. / Study of the animal performance and herbage production of irrigated tanzania grass (Panicum maximum, Jacq.) in a rotational grazing system under three post-grazing residues levels.Marco Antonio Penati 02 April 2002 (has links)
O objetivo dessa pesquisa foi o de avaliar o efeito de massa de forragem no resíduo sobre as características morfológicas, produção das pastagens, desempenho e produtividade animal. O experimento foi desenvolvido no período de 30 de outubro de 1999 até 03 de janeiro de 2001 em um sistema rotacionado de pastagens irrigadas de Panicum maximum Jacq. cv. Tanzânia estabelecidas na Escola Superior de Agricultura "Luiz de Queiroz" em Piracicaba/SP. O delineamento experimental foi o de blocos (4) completos ao acaso com três disponibilidades de forragem no resíduo (1000, 2500 e 4000 kg/ha de matéria seca verde (MSV)). A semeadura foi realizada em 17 de março de 1999 e a adubação de correção do solo tinha o objetivo de atingir os seguintes valores: 80% de saturação por bases; 30 mg/cm3 de fósforo e 5% de potássio na CTC. Os piquetes foram adubados com 334 kg/ha/pastejo de 24-04-24. Os animais utilizados foram machos castrados da raça nelore com cerca de 10 meses de idade. Foram conduzidos quatro pré experimentos sendo três deles para deteminar a melhor relação dimensão: formato da unidade amostral e a quantidade de amostras para avaliar a massa de forragem. O quarto pré experimento teve como objetivo avaliar a relação entre profundidade de pastejo e altura das planta antes do pastejo. A avaliação da massa de forragem em piquetes estabelecidos com plantas que formam touceiras deve ser realizada através de quatro amostras de 1 m2 (1m x 1m). A profundidade de pastejo foi relacionada linearmente com a altura das plantas independente da intensidade de pastejo e do dia de ocupação do pasto. A intensidade de pastejo não teve efeito sobre o angulo de inclinação dos perfilhos mais externos da touceira, sobre o formato das touceiras e sobre a distribuição da densidade de forragem no perfil da pastagem indicando que o capim Tanzânia tem pouca plasticidade em relação a essas características. Entretanto a proporção de haste, de folhas, de material morto, da altura, da relação folha/haste foram características morfológicas afetadas pelos tratamentos e épocas do ano. A taxa de cobertura do solo diminuiu durante período experimental independente do tratamento enquanto que o tamanho das touceiras esteve associado a mecanismos rápidos de adaptação da planta a mudanças no meio ambiente causadas pelas diferentes disponibilidades de forragem. O desempenho animal esteve mais relacionado com a oferta e disponibilidade de forragem do que com a relação folha/haste e a densidade de forragem. Apesar da produtividade animal não ser alterada pelos tratamentos, o manejo mais adequado seria o de manter a disponibilidade de forragem no resíduo superior a 2915 kg MSV/ha assim como as ofertas de forragem maiores do que 14,9 kg MSV/100 kg PV ou 6,1 kg de folhas/100 kg PV afim de melhorar o desempenho dos animais e reduzir a idade de abate em 150 dias quando comparado com o tratamento que teve como massa de forragem no resíduo 1266 kg MSV/ha. / This research was to evaluate effect of the post-grazing pasture mass on morphological characteristics, herbage production and animal performance and productivity from irrigated and rotationally grazed system with Panicum maximum Jacq. cv. Tanzânia. The experimental period was October 30/1999 to January 03/2001 and established at Escola Superior de Agricultura "Luiz de Queiroz", Piracicaba/SP. A complete randomized block design with three postgrazing residues (1.000, 2.500 e 4.000 kg/ha of green dry matter - GDM) and four replicates was used. The grass seeding was performed on March 17/1999 and soil was limed and fertilized to 80% base saturation, phosphorus (resin) level of 30 mg/cm3 and 5% potassium in the cation exchange capacity. In each grazing cycle paddocks were fertilized with 24-04-24 (334 kg/ha). Nellore steers with approximately 10 months of age were used. Four pre-experiments were conducted, the first three to determine the best dimesion:format ratio of the sampling unit and the most appropriate number of samplings in evaluating herbage mass. The fourth pre-experiment aimed to evaluate the relationship between grazing depth (progressive sward defoliation during the grazing period) and plant height before grazing. Herbage mass evaluation in tussock-forming pastures should be performed taking four 1 m2 samples (1m x 1m). Grazing depth was linearly correlated to plant height, irrespective of grazing intensity and days on the grazing period. Grazing intensity had no effect on the slope of the angle between soil and tillers on the border of the tussock, neither on tussock shape nor on forage bulk density across the sward profile, indicating Tanzania grass has limited plasticity for these characteristics. However, stem, leaf and dead material proportion in herbage samples, as well as leaf/stem ratio and plant height were affected by treatments (post-grazing residues) and season of the year. The ground area covered by grass tussocks decreased during the experimental period, irrespective of the treatment. The tussock size was associated with fast adaptation mechanisms of the plants in response to changes in environment resulting from different post-grazing herbage masses. Animal performance was better related to herbage allowance and post-grazing residues than with leaf/stem ratio or forage bulk density. Treatments did not altered animal productivity. In spite of this, it would be more adequate to allow post-grazing pasture mass to be higher than 2.915 kg GDM/ha and herbage allowance to be higher than 14,9 kg GDM/100 kg live weight (LW) or 6,1 kg of leaves/100 kg LW. This grazing management would improve animal performance and anticipate slaughter age in 150 days when compared to the treatment with post-grazing pasture mass of 1.266 kg GDM/ha.
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Análise de um experimento de seleção para crescimento em bovinos nelore: respostas direta no peso ao sobreano e correlacionadas no tamanho e reprodução das matrizes. / Analysis of a selection experiment for growth in nelore cattle: direct response of yearling weight and correlated responses of size and reproduction of cows.Maria Eugênia Zerlotti Mercadante 12 December 2001 (has links)
Foram estudados dados provenientes do experimento de seleção estabelecido na Estação Experimental de Zootecnia de Sertãozinho (IZ - SP), com o objetivo de avaliar a resposta direta no peso ao sobreano de machos e fêmeas e as respostas correlacionadas no tamanho e caracteres reprodutivos das matrizes. O experimento iniciou em 1978, com três rebanhos da raça Nelore, seleção (NeS), tradicional (NeT) e controle (NeC), sendo os dois primeiros selecionados para maior peso ao sobreano e o terceiro para a média deste peso. O NeT diferencia-se de NeS por ter recebido, eventualmente, touros de outros rebanhos, inclusive comerciais. Registros de dias ao parto foram analisados separadamente nas novilhas, vacas e vacas e novilhas sem incluir e incluindo as fêmeas não paridas. Nenhuma diferença significativa (P<0,05) foi detectada entre os desempenhos das fêmeas dos rebanhos selecionados (NeS e NeT) e as do controle (NeC). As herdabilidades variaram de 0,02 a 0,16, sendo as mais altas obtidas em arquivos nos quais foram incluídos os registros das fêmeas não paridas. Os registros repetidos de dias ao parto de vacas e novilhas incluindo as fêmeas não paridas foram também analisados por modelo de regressão aleatória com polinômios ortogonais (Legendre) da idade na monta (em anos) como a covariável independente, para avaliar este caracter longitudinalmente durante a vida produtiva das fêmeas. As herdabilidades estimadas aumentaram de 0,08 a 0,28 da 1 a à 6 a monta. A seleção para peso não alterou o valor genético médio das vacas de NeS e NeT, entretanto os valores genéticos das vacas do NeC mostraram tendência de queda no decorrer dos anos, além de longitudinalmente, com o avanço da idade. Registros de peso ao sobreano dos animais nascidos de 1978 a 1998, e da altura na garupa ao sobreano das fêmeas nascidas a partir de 1985 foram analisados por modelo animal para obter a tendência genética de 1981 a 1996. Médias de peso, altura e escore de condição corporal na entrada da monta, dias ao parto e sucesso ao parto das matrizes nascidas nos últimos 4 anos foram contrastadas entre os selecionados e o NeC, em análise de quadrados mínimos. As tendências genéticas médias obtidas em 16 anos foram 1,7±0,2; 2,3±0,2 e 0,1±0,1 kg/ano nos machos e 1,9±0,2; 2,4±0,2 e 0,1±0,1 kg/ano nas fêmeas para peso, e 0,25±0,03; 0,24±0,04 e 0,04±0,03 cm nas fêmeas para altura ao sobreano, respectivamente em NeS, NeT e NeC. Novilhas e vacas de NeS e NeT foram 19% e 15% mais pesadas e 4% mais altas na entrada da monta, em relação às do NeC. Não foram detectadas diferenças significativas (P<0,05) no desempenho reprodutivo das matrizes selecionadas em relação ao NeC. Os resultados indicaram que a seleção para peso corporal promoveu respostas altas e consistentes nos pesos e alturas ao sobreano e em idades mais tardias, sem comprometer o desempenho reprodutivo das matrizes. / Data from a selection experiment, carried out AT Experimental Station of Sertãozinho, State of São Paulo, Brazil, were analyzed in order to evaluate the direct response of yearling weight and the correlated responses of the size and reproduction traits of cows. The experiment started in 1978, with three lines of Nelore cattle, selection (NeS), traditional (NeT), both selected for greater yearling weight, and control (NeC), selected for mean yearling weight. The NeT line is different from the NeS one because sires from another herds were eventually introduced into the NeT line. Records of days to calving were analyzed separately for heifers, cows and cows and heifers with and without non-calver females. There were no significant differences (P<0.05) between the performance of females from the selection lines (NeS and NeT) and those from the control line (NeC). The heritabilities were between 0.02 and 0.06, and the higher estimates were from data sets including non-calver females. The repeated records of days to calving of cows and heifers with non-calvers, were too analyzed using a random regression model, with orthogonal polynomials (Legendre) of age in the breeding season (in years) as the independent covariable, to order to evaluate this trait longitudinally during the herd life of the females. The estimates of heritability increased from 0.08 to 0.28, from first up to 6 th breeding season. The selection for weight did not alter the average breeding value of cows from NeS and NeT, although the average genetic breeding values of cows declined throughout the years, and too longitudinally, with advancing age. Yearling weight of the animals born from 1978 to 1998, and the yearling hip height of females born from 1985 to 1998, were evaluated using an animal model, in order to obtain the genetic trends for the years 1981 to 1996. The averages for weight, height and body condition score at the start of the breeding season, days to calving and calving success of cows born in the last 4 years, were contrasted between the selected (NeS and NeT) and control lines. The genetic trends obtained after 16 years were 1.7±0.2; 2.3±0.2 and 0.1±0.1 kg/year for males and 1.9±0.2; 2.4±0.2 e 0.1±0.1 kg/year for females with respect to weight, and 0.25±0.03; 0.24±0.04 and 0.04±0.03 cm for females with respect to yearling height for the NeS, NeT and NeC lines, respectively. Heifers and cows from NeS and NeT were 19% and 15% heavier and 4% taller at the start of the breeding season than those from NeC. Significant differences (P<0.05) were not detected with respect to the reproductive performance between selected (NeS and NeT) and control females. The results indicated that the selection according to body weight gave high and consistent responses for weight and height both at the yearling and later ages, without compromised the reproductive performance of the cows.
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Efeitos de diferentes taxas de crescimento na recria sobre o desempenho, idade a puberdade e produção leiteira em fêmeas da raça Santa Inês / Effects of growing rates in prepubertal period on performance, age at puberty and milk yield of Santa Ines sheepMarlon Richard Hilário da Silva 30 November 2009 (has links)
Experimento I: Foram utilizadas 60 borregas da raça Santa Inês com o objetivo de avaliar os efeitos de três taxas de crescimento até os 7 meses de idade. Os tratamentos foram delineados visando obter alta taxa de crescimento (ATC), moderada taxa de crescimento (MTC) e baixa taxa de crescimento (BTC). Foram avaliados o consumo de matéria seca, o ganho de peso corporal e as concentrações séricas de progesterona para a detecção da idade à puberdade. Foram verificados efeitos lineares (p<0,01) para o peso corporal e metabólico, sendo os maiores valores encontrados para os animais do tratamento ATC. Efeito linear (p<0,01) foi verificado para o ganho médio diário (GMD) sendo de 0,182; 0,156 e 0,136 kg/dia para os tratamentos ATC, MTC e BTC respectivamente. Borregas pertencentes ao tratamento ATC apresentaram maior conversão alimentar sendo os valores para o tratamento ATC de 6,1; MTC de 7,1 e BTC de 9,2 kg MS/ kg de ganho de peso. Verificou-se diferença (p<0,01) quanto a porcentagem de animais que atingiram a puberdade sendo o tratamento MTC com o maior número de borregas púberes até os 7 meses de idade (60%), seguida dos tratamentos ATC (45%) e BTC (15%)com peso médio de 39,3 kg de peso corporal, sendo 70% do peso adulto. Experimento II: As borregas utilizadas no experimento 1 com 37 Kg de PC entraram em estação de monta e após o parto receberam uma única dieta, sendo blocadas de acordo com o peso e dias de lactação. Observou-se efeito quadrático (p<0,01) na produção de leite em 3h (137,9; 150,1 e 104,4 g), efeito quadrático (p<0,05) para LCG (167,2; 178,9 e 129,4) e efeito quadrático (p<0,03) LCGP (216,5; 140,5 e 162,7). Os teores de gordura (9,8; 8,9 e 8,3%) e de proteína (4,2; 4,2 e 4,6 apresentaram efeitos lineares sendo (p=0,01) para gordura e (p<0,01) para proteína. Experimento III: Os objetivos deste experimento foram avaliar o efeito da inclusão de bagaço de cana-de-açúcar in natura (BIN) sobre o consumo e a digestibilidade aparente dos nutrientes, balanço de nitrogênio e os parâmetros ruminais de cordeiros Santa Inês. Quatro cordeiros providos de cânula ruminal com peso corporal (PC) médio inicial de 30kg foram alojados individualmente em gaiolas para ensaios de metabolismo. O tratamento controle positivo (CONT) consistia de uma ração de alto concentrado contendo de 10% de feno de coastcross (Cynodon sp.) na MS, sendo os demais tratamentos níveis crescentes de BIN em 10, 20 e 30% da MS, constituindo os tratamentos 10BC, 20BC e 30BC. As rações experimentais foram isonitrogenadas (13,75±0,25% PB). Não verificou-se efeito de tratamento (p>0,05) sobre o consumo de MS, MO, e PB em kg/dia e g/kg de PC 0,75; apenas sobre consumo de FDN com efeito linear (p< 0,01). O balanço de nitrogênio, assim como a concentração de acetato, propionato, butirato, concentração total de AGCC, amônia no fluido ruminal e a de digestibilidade aparente da MS, MO, PB e FDN não foram afetados pelos tratamentos. / Trial I: Sixty Santa Ines ewe lambs were used to evaluate the effects of three growing rates until 7 months of age. Treatments were defined to achieve high (HGR), moderate (MGR) and low (LGR) growing rates. The dry matter intake and average daily gain (ADG) were evaluated, well as serum levels of progesterone in order to detect the age of puberty. Linear effects (P<0.01) to body weight (BW) with values being 59.5, 61.4, and 58.0 to HGR, MGR and LGR, respectively. Linear effects (P<0.01) was also observed for average daily gain with values being 0.182, 0.156, and 0.136 kg/day, for HGR, MGR and LGR, respectively. The feed:gain ratio showed a linear effects (p<0.02) according to raw sugarcane bagasse inclusion in diet with values being 6.1, 7.1 (M) and 9.2 kgDM/kg BW for HGR, MGR and LGR, respectively. Linear effects (p<0.01) was observed for the percentage of animals of ewe lambs the reached puberty at 7 months of age, showing highest values for MGR (60%), followed by HGR (45%) and LGR (15%) treatment. Trial II: The same ewe lambs of Experiment 1 were bred at 37 kg of BW and after lambing they received a unique diet. The ewe lambs were blocked according to body weight and days in milking. According to the energy density increase in the diet, quadratic effects (p<0.01) was observed on milk production recorded in a 3-hour interval (137.9; 150.1 and 104.4 g), fat corrected milk (167.2, 178.9 and 129.4 g) and fat and protein corrected milk (216.5, 140.5, 162.7g). Fat (9.77, 8.92 and 8.35%) and protein (4.2, 4.2, and 4.6%) showed linear effects (P=0.01) for fat and (P<0.01) for protein. Trial III: The aim of this study was to evaluate the inclusion of raw sugarcane bagasse (RSB) on dry matter intake and apparent digestibility of nutrients, nitrogen balance and ruminal parameters of Santa Ines ram lambs. Four ruminally fistulated ram lambs averaging 30 kg of BW were placed in metabolism crates. The control treatment (CONT) consisted of a high concentrate diet with 10% of coastcross (Cynodon sp.) hay in the dietary DM and the others treatments consisted of 10, 20 and, 30% of RBS in the dietary DM. The diets were formulated to provide similar concentration of crude protein (13.75±0.25%). No effects were observed for DMI, OM, and CP in kg/day and g/kg of BW0.75. Linear effects (p<0.02) was observed for NDF intake according RBS inclusion. Variables, as well as total SCFA, acetate, propionate, and butyrate concentrations. Ruminal ammonia and apparent nutrient digestibility also did not differ among treatments.
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Unlocking the role of small heat shock proteins and apoptosis in postmortem proteolysis and meat quality characteristics of skeletal muscles under different conditionsDanyi Ma (8202711) 28 April 2020 (has links)
<p>Postmortem aging has been
extensively practiced as value-adding process due to the beneficial impacts on
meat palatability. Meat tenderization occurred through proteolytic
fragmentation of myofibrillar structural proteins via endogenous protease
systems, which is considered as the primary drive to enhance major palatability
attributes including tenderness, juiciness, and flavor. Recent theoretical
framework proposes apoptosis, or programmed cell death, as the preceding step
that initiates postmortem proteolysis. Whereas small heat shock proteins have
been consistently recognized as meat quality biomarkers, probably due to their
protective activities against proteolysis through anti-stress, anti-apoptotic,
and chaperoning functionalities. To shed light on detailed mechanisms
controlling postmortem proteolysis and consequential impacts on the development
of fresh meat quality characteristics, postmortem proteolytic changes of small
heat shock proteins, apoptotic factors, and myofibrillar structural proteins
were profiled in postmortem skeletal muscles under different metabolic
backgrounds and across species. </p>
<p>In beef, three muscles, <i>longissimus
lumborum</i> (LL), <i>semimembranosus</i> (SM), and <i>psoas major</i> (PM),
have been selected to represent glycolytic, intermediate, and oxidative muscle
types. Tenderness and water - holding capacity were determined, and
proteolysis, apoptotic features, and small heat shock proteins were measured in
8 beef carcasses at 1, 2, 9, 16, and 23 days of aging. PM exhibited limited
aging potential in quality developments shown by lower extents of shear force,
water-holding capacity, and proteolytic changes, including calpain 1 autolysis,
troponin T, and HSP27 compared to LL and SM. Conversely, LL had an increase in
tenderization and water-holding capacity, which was accompanied with more
extended calpain 1 autolysis, proteolysis and HSP27 degradation, compared with
other muscles. The results of this study suggest that postmortem proteolytic
changes of myofibrillar proteins, small HSPs and apoptotic factors occur in a
muscle-specific manner, which is likely attributed to different rate and extent
of meat quality developments of each muscle during aging. </p>
<p>Callipyge lambs are a unique
genetic background showing calpastatin over-expression, muscle hypertrophy in
loin and hindquarter area, substantially compromised meat tenderization
potential, and a shift of muscle fiber composition towards fast-glycolytic
directions. Proteome and metabolome changes in muscles from callipyge mutation
(+/C) and non-callipyge phenotype (+/+, C/+, and C/C) lambs were profiled to
provide insight into the biochemical changes affecting meat quality attributes.
M. longissimus thoracis from lambs with all four possible callipyge genotype (n
= 4, C/+, C/C, +/C, and +/+) were collected after 3d aging and analyzed using
mass-spectrometry based platforms. Among identified proteomes, cytochrome c (pro-apoptotic
protein) was detected with significantly lower abundances in +/C.
Anti-apoptotic HSP70, BAG3, and PARK7 were over-abundant in +/C, which could
result in delayed apoptosis and possibly attributed to tougher meat in
callipyge lambs. Eight glycolysis enzymes were overabundant in +/C lambs,
whereas 3 enzymes involved in TCA cycle were overabundant in non-callipyge ones
(C/C and/or C/+). Twenty-five metabolites were affected by genotypes (P <
0.05), including metabolic co-factors, polyphenols, and AA/short peptides.</p>
<p>Pig production is facing increased
public pressure regarding antibiotic usage restriction. Recently, dietary
L-glutamine at cost effective level (0.2%) was identified as an effective antibiotic
alternative in post-transport nursery pig diets. To evaluate carcass and meat
quality characteristics in market-ready pigs when 0.2% dietary L-glutamine was
applied as for early-life post-weaning and transport recovery, pigs (N=480)
were weaned and transported in two replication trials in SPRING (April of 2017)
vs. SUMMER (July of 2016), fed 3 different diets (Non: no antibiotic, Anti: 441
ppm chlortetracycline and + 38.6 ppm tiamulin, Gln: 0.20% L-glutamine) for 14
days after transport, and fed basal diet until reaching market weight. Pairs of
<i>longissimus dorsi</i> (LD) and <i>psoas major</i> (PM) muscles from each
carcass (n=10/diet/trial) were separated at 1 d and 7 d postmortem,
respectively. Carcass yield and meat physical and quality attributes were
evaluated. Overall impacts of Gln on physical attributes of carcasses and
porcine muscles were minimal. No dietary effects were found in carcass,
proximate composition, water-holding capacity, or shear force. Significant
difference between trials were found in terms of productivity and pork/carcass
qualities, where SPRING replicates showed increased body weight, faster pH
decline, paler surface color, higher intra-muscular fat deposition, and
improved tenderness and water-holding capacity as indicated by lower shear
force values, thaw-purge loss, and cooking loss (P < 0.05).</p>
<p>The pork and carcass quality
results give rise to a postulation that different metabolism and animal growth
might have been occured between the two production trials, consequentially
differentiated meat quality development. In this regard, myofibrillar
proteolysis, small heat shock proteins, and apoptotic factors were characterized
during 7 d postmortem aging in porcine LD and PM muscles from both seasonal
trials, combined with metabolomics profiles of 1d samples using the
GC-TOF-MS/MS platform. Compared to SUMMER counterparts, SPRING muscles showed
concurrence of more extended apoptosis, further calpain 1 autolysis, and
increased structural protein degradation (P<0.05). SPRING muscles showed
more ATP catabolism compounds and increase in carbohydrates, branched-chain
amino acids, and 16-18 carbon fatty acids, which could be chemistry
fingerprints of increased cellular oxidative stress, consequentially favoring
onset of apoptosis and proteolysis. Meanwhile, SUMMER pigs showed increased
stress-defending metabolites, such as ascorbic acid, antioxidant amino acids,
and decreased inhibitory neuro-transmitter GABA, which may indicate elevated
stress-defending activity in SUMMER pigs that possibly inhibited apoptosis and
proteolysis. </p>
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December 2022 Final Thesis. G. Ceja..pdfGuadalupe Ceja (14216219) 07 December 2022 (has links)
<p>(From abstract) </p>
<p>In the first study, the urine collection method was effectively applied for evaluation of intestinal permeability using Cr-EDTA, an indigestible oral marker, demonstrating the applicability of the procedure in 1-week-old and 6-week-old neonatal heifer calves (n=15 calves). Calf health observations were recorded during the entire urinary catheterization process and collection period to evaluate any negative health reactions to the procedure, or localized reactions. Proportion of localized reactions were analyzed, and the proportions did not exceed 20% for the calves catheterized at either 1 week or 6 weeks of age. </p>
<p>In the second study, the developed catheterization procedure and urine collection method was applied using Cr-EDTA as an oral marker to investigate if L-GLN supplementation would offer improvement to intestinal permeability. In this larger study, 30 Holstein heifer calves [1.5 ± 0.5 days old; 37.1 ± 0.86 kg body weight (<strong>BW</strong>)] were blocked by serum total protein, BW, and age, and randomly assigned to 1 of 2 treatments: <strong>GLN</strong> [24% crude protein (<strong>CP</strong>)], 17% fat milk replacer (<strong>MR</strong>) +10 g L-GLN/kg MR powder) or <strong>NS</strong> (24% CP, 17% fat MR). MR was reconstituted to 12.5% solids with warm water and fed 3.8 L/calf/d until weaning. Calves were weaned at 56.4 ± 0.5 days of age, and had <em>ad libitum</em> grain (17% CP, 2% fat) and water access throughout the experimental period.</p>
<p>During the preweaning period, calves were individually housed in hutches and health observations, which included respiratory and fecal scores, were assessed daily. Body weight was measured weekly, and grain and MR intake was assessed daily to calculate average daily gain (<strong>ADG</strong>), average daily feed intake [<strong>ADFI</strong>; grain intake (dry matter (<strong>DM)</strong> basis) + MR intake (DM basis)], and feed efficiency (<strong>G:F</strong>; ADG:ADFI). At weaning, calves were weighed, moved to pens (n = 3 pens/treatment, 4-5 calves/pen), provided free access to grain and grass hay, and then weighed 2 weeks post-weaning. Additionally, urinary catheters were placed at 1 and 6 weeks of age, and calves were orally dosed with 1 L Cr-EDTA in their MR. Urine samples were then collected over a 24-hr period for Cr output analysis as an <em>in vivo</em>biomarker of intestinal permeability. </p>
<p>Blood was collected on study days 1, 2, 5, 7, 14, 21, 42, 56, and 70 to measure haptoglobin, serum amyloid A, leukocyte data, neutrophil: lymphocyte (<strong>N:L</strong>), glucose, non-esterified fatty acids, insulin, and cortisol. Two study periods were identified for data analysis representing greater (<strong>P1</strong>; weeks 1-3) and reduced (<strong>P2</strong>; weeks 4-8) enteric disease susceptibility. Data were analyzed using PROC GLIMMIX or PROC MIXED in SAS 9.4 with calf as the experimental unit. There was a decrease in total preweaning Cr output (<em>P</em> < 0.05) for GLN calves, and Cr output in 1 week old calves was decreased (<em>P</em> = 0.04) in GLN versus NS calves. The N:L was decreased overall (<em>P</em> = 0.03) and during P2 (<em>P</em> = 0.01) and P2 neutrophil count tended to be reduced (<em>P</em> = 0.07) in GLN versus NS calves. There were no MR treatment differences for ADFI, ADG, body measurements, post-absorptive metabolic biomarkers, disease scores, and therapeutic treatments (<em>P</em> > 0.10). In summary, L-GLN supplementation improved intestinal integrity and biomarkers of physiological stress in pre-weaned Holstein heifer calves managed under production-relevant conditions. </p>
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Great Eggs-pectations: Understanding Markers of Stress and Welfare in Pullets Using Feeder Space and Stocking Density StressorsMeagan E Abraham (14206337) 09 December 2022 (has links)
<p> </p>
<p>Little is known about pullet stress and welfare and even less is known about pullets in cage-free housing. The studies included in this dissertation evaluated bird responses to stocking density, feeder space, and ACTH stressors in pullets. The project and procedures were reviewed and approved by the Purdue University Animal Care and Use Committee (Protocol#:2001002002 and # 1908001938). </p>
<p>Two studies of external stressors and one study of an internal stressor were applied to Lohmann LB-Lite (Brown) and Lohmann LSL-Lite (White) pullets. For both studies of an external stressor, bird-based welfare parameters included evaluation of bird appearance through feather coverage, foot condition, and keel condition; blood parameters, i.e., cholesterol, corticosterone, cortisol, NDV/IBV IgG/IgY titer, and differential WBC (%) and heterophil to lymphocyte ratio; measurements of the spleen, bursa, thymus, and liver; and production values measured through body weight, body weight uniformity, shank length, feed consumption, and feed conversion ratio (FCR). Birds were followed from 2-16 weeks of age (WOA) for the external stressors and were housed on the floor.</p>
<p>The first external stressor was applied through a 2 x 2 factorial randomized complete block design (RCBD) using Brown and White strains and two stocking densities- high (HSD) and low (LSD). Stocking densities were adjusted over time, with the ending density of 670 cm2/bird for HSD and 1352 cm2/bird for LSD at 16 WOA. Density alone affected relative bursal weight (%), with LSD generating heavier bursas. LSD also generated lower, better FCR for both white and brown strains though this difference was not statistically significant. Within a density, opposing strain effects were seen for body weight uniformity and feather coverage. White was improved at HSD while Brown was improved for LSD. </p>
<p>The second external stressor study was a 2 x 2 x 3 factorial RCBD with Brown and White, 2 feeder space allocations (3.5 or 7 cm/bird), and 3 stocking densities. The ending stocking densities were 154.45, 518.76, and 1,247.38 cm2 for the high (HSD), medium (MSD), and low (LSD) stocking density treatments, respectively, at 16 WOA. In this study, less space per bird was associated with poorer feather condition but improved FCR. Reduced feeder space was associated with elevated or worse FCR.</p>
<p>In a study of internal stressors, ten each of Lohmann LSL-Lite hens received either a saline-mannitol or synthetic human ACTH injection at 6, 17, and 26 weeks of age. Blood was collected prior to injection and 1- and 2-hours post-injection. H:L ratio, WBC differential counts, corticosterone, and cortisol were measured in blood. Albumen samples were collected to measure corticosterone and cortisol at 26 WOA only but were unaffected by treatment. Serum corticosterone and H:L ratio were also unaffected by treatment. Serum cortisol was increased 1 hour after ACTH injection at 17 and 26 WOA and percent heterophils and lymphocytes were affected by ACTH at 26 WOA indicating potential utility as a measure of stress for adult birds only. </p>
<p>The results of these studies show that pullets are resilient and resistant to several stressors. Both acute and chronic as well as internal and external stressors were tested in these studies with limited effects on the birds’ physiology, immunity, condition, or production. Feather coverage and bursal size were improved at lower stocking densities and FCR was improved at greater feeder space but the implications of these differences are unclear. Further research is needed to identify why there are reduced stress responses in pullets; it is possible that domestication or preserved evolutionary pathways explain some of the differences between pullet and adult stress responses. And while the majority of parameters were unaffected by the stressors used, the results of this study presented baseline values for Lohmann LB-Lite and Lohmann LSL-Lite pullets that will help guide future research. Additionally, the results of these studies left perhaps more questions than answers but illustrate perfectly that more research will be critical to an understanding of pullet stress and welfare.</p>
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<b>Investigating epigenetic mechanisms in early porcine development</b>Sarah M Innis (18239221) 22 March 2024 (has links)
<p dir="ltr">Epigenetics involves the study of mechanisms that influence gene expression. These mechanisms are heritable and dynamic, and despite altering gene transcriptional activity, they do not change the underlying DNA sequence. While epigenetic mechanisms govern gene expression throughout the lifetime of an organism, the dynamic nature and precision of the transcriptional control afforded by processes such as histone modifications and chromatin architecture remodeling are exemplified in early mammalian development. Perhaps unsurprisingly, perturbations to the epigenetic status of a cell can alter its function, and widespread epigenome disruptions due to changes in the developmental environment can compromise the growth, and even viability, of an embryo or fetus. By studying epigenetic mechanisms and the patterns they impart, we can better understand not only how developmental progression is regulated during embryonic development and beyond, but also what the consequences of aberrant epigenetic disturbances may be to developing organisms.</p><p><br></p><p dir="ltr">Many gaps remain in our knowledge concerning epigenetic mechanisms in domestic livestock species, particularly regarding early development. Pigs represent a compelling model organism for study in this area, as they are increasingly being used as a biomedical model for human-oriented research due to their physiological similarities to humans, and they remain a staple meat species in many parts of the world. Chapter 2 investigates the presence and transcriptional dynamics of the SWI/SNF chromatin remodeling complex GBAF in porcine trophectoderm (PTr2) and fetal fibroblast (PFF) cells. These cell lines represent two discrete developmental stages during early swine development, with the PTr2 cells originally obtained from the trophectoderm of a gestation day 12 elongating porcine conceptus, and the fetal fibroblast cells were collected from a fetus on day 40 of gestation. Using immunocytochemistry and Western blotting techniques, GBAF was identified in both cell lines. Further, co-immunoprecipitation of GBAF constituent subunits and other BAF family subcomplex subunits revealed a previously undescribed interaction between the GBAF subunit GLTSCR1 and the BAF subunit BAF170, the latter of which has not been shown to be present in human and mouse GBAF. This may suggest a species-specific GBAF composition in swine. Analysis of RNA-seq data from porcine embryos, PTr2 cells, and PFF cells showed that while transcription of GBAF-specific subunits BRD9 and GLTSCR1 was detectable, expression levels were lower compared to other BAF family subunits. Taken together these results suggest that, while GBAF is detectable in swine early development contexts, it may have a comparatively minor contribution as an epigenetic mechanism during the represented developmental timepoints.</p><p><br></p><p dir="ltr">Details in the literature about the epigenetic landscape and the resulting chromatin state during porcine early development are also limited at present. Chapter 3 involves the global epigenetic profiling of the histone marks H3K4me3, H3K27ac, and H3K27me3 and the SWI/SNF central ATPase BRG1 in PTr2 and PFF cells using CUT&RUN. The enrichment patterns observed for these features were consistent with known patterns described in the literature. H3K4me3 was primarily enriched in gene promoter regions, and H3K27ac showed enrichment in both promoter regions and distal intergenic regions, some of which are likely active enhancers. H3K27me3 showed broad genomic localization and was detected at genes known to be transcriptionally inactive in these cell types, as well as in distal intergenic regions. BRG1 showed some co-enrichment with H3K4me3 and H3K27ac in promoter regions, as well as several instances of H3K27ac co-enrichment at intergenic sites. The sequencing files were used to build a chromatin state prediction model for 10 states in each cell line, ranging from TSS to repressed genomic regions. Additionally, the transcriptomes of PTr2 and PFF cells were compared to those of human cells taken from comparable gestational time points to determine if these swine cell lines could potentially serve as translational <i>in vitro</i> models. PTr2 cells and human trophectoderm (TE) cells were relatively dissimilar in their cell-type specific gene identities (~24% overlap) and corresponding transcriptional levels, but the porcine and human fibroblast cells shared around 50% of the same cell type-specific genes, and expression levels were broadly similar among them. Altogether, these findings provide foundational epigenetic landscape information for PTr2 and PFF cells and potential insights regarding similarities and differences in cell identity between human and pig trophectoderm and fetal fibroblast cells.</p><p><br></p><p dir="ltr">The placenta is a transient organ that provides essential support to the developing fetus in the form of nutrient and gas exchange. Despite its significance in facilitating fetal development, our understanding of how the placenta is affected by its environment is greatly limited, and only a handful of studies exploring the placental epigenome in swine exist to date. To address these gaps, and building upon the epigenetic profiling methods developed in Chapter 3, Chapter 4 investigated whether, and to what extent, the placental epigenome changes in response to fetal endocrine perturbations. Placental tissue was collected at day 86 of gestation from untreated pregnant gilts and pregnant gilts treated for 21 days with methimazole (MMI) to induce fetal hypothyroidism. CUT&RUN was used to evaluate the enrichment of H3K4me3, H3K27ac, and BRG1 in placental tissue derived from n=6 male and female fetuses in each treatment group (n=12 samples per group). Differential enrichment of all three epigenetic features was seen in placental tissues obtained from MMI-treated fetuses, and, notably, existing sex-specific differences in placental epigenetic features were exacerbated by MMI-induced fetal hypothyroidism. This may suggest that the porcine placenta may be impacted by fetal endocrine status during late gestation. Together, these findings show that sex-specific differences in placental chromatin state exist and that a fetal hypothyroid state is sufficient to perturb the placental epigenome, ultimately providing novel insights into the intricate interplay between fetal endocrine status and regulation of the placental epigenome.</p>
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Desempenho, características de carcaça e qualidade de carne de bovinos de diferentes grupos genéticos suplementados com ácidos graxos poli-insaturados / Performance, carcass traits and meat quality of cattle from different genetic groups supplemented with polyunsaturated fatty acidsAntonelo, Daniel Silva 20 June 2018 (has links)
Este trabalho foi desenvolvido para avaliar o efeito da suplementação com ácidos graxos poli-insaturados (AGPI) sobre o desempenho, características de carcaça e qualidade de carne de bovinos de diferentes predisposições genéticas para deposição de gordura intramuscular (GIM). Foram utilizados 60 machos não-castrados, dos quais 30 Nelore (NE) e 30 cruzados Nelore x Aberdeen Angus (NA), com uma média de 363 ± 28 kg de peso vivo inicial e 24 meses de idade. Os animais foram designados a um delineamento em blocos casualizados (peso vivo inicial) com um arranjo fatorial 2 x 2 (dieta e grupo genético), usando 15 repetições por tratamentos. Os animais foram alimentados durante 133 dias com uma das seguintes dietas: CO - dieta controle, sem adição de óleo de soja; OS - dieta controle contendo 3,5% de óleo de soja em substituição ao milho grão moído. Foi analisado o ganho médio diário (GMD), a eficiência alimentar (EA), a ingestão de matéria seca (IMS) e o peso vivo animal. No abate, foi avaliado o peso (PCQ) e o rendimento (RCQ) de carcaça quente, bem como a área de olho de lombo (AOL), a espessura de gordura subcutânea (EGS) e o escore de marmorização da carne. Amostras do músculo Longissimus foram utilizadas para avaliação da força de cisalhamento (CS), comprimento de sarcômero, análise sensorial e metaboloma da carne, bem como a vida útil e oxidação lipídica e o perfil de ácidos graxos e sua implicação na saúde humana. Todos os dados foram analisados utilizando o procedimento MIXED do software SAS. Não houve interação entre dieta e grupo genético para a maioria das características avaliadas, de modo que seus efeitos podem ser considerados separadamente. Animais alimentados com a dieta OS apresentou maior GMD (P = 0,0392) e EA (P = 0,0020) sem modificar a IMS e as características de carcaça quando comparado aos alimentados com a dieta CO. Não houve efeito da dieta em relação à FC e a cor da carne, porém a carne dos animais alimentados com OS apresentaram menores notas de aceitação geral (P < 0,0001), maciez (P < 0,0001) e suculência (P = 0,0009), além de uma tendência de menores notas de sabor (P = 0,0533) quando comparados aos animais CO. Não houve efeito da dieta sobre a quantidade de GIM, porém a carne de animais alimentados com a dieta OS tiveram menor proporção dos ácidos graxos da família n3 (P = 0,0287), reduzindo também a relação n-6:n-3 (P = 0,0256) e o índice de qualidade (P = 0,0133) da carne para a saúde humana. Em relação ao grupo genético, os animais NA tiveram maior IMS (P = 0,0479) e tenderam a ter maior GMD (P = 0,0907) e peso vivo ao abate (P = 0,0596) do que animais NE, sem alterar a EA e qualquer característica de carcaça. Não houve efeito do grupo genético em relação à FC, porém a carne dos animais NE apresentaram maiores notas de aceitação geral (P = 0,0440) e sabor (P = 0,0444) do que as dos animais NA. A carne de animais NE apresentaram maiores valores de L* (P = 0,0003) e b* (P = 0,0023) e, consequentemente, maiores valores de croma (P = 0,0358) e ângulo de inclinação (P = 0,0073) comparados à carne de animais NA. Não houve efeito do grupo genético sobre a quantidade de GIM, além de nenhuma modificação na proporção de ácidos graxos. Além disso, o metaboloma da carne foi modificado (P < 0,10) tanto pela dieta quanto pelo grupo genético em 8 e 15 metabólitos, dos 31 identificados, respectivamente. Isoleucina, glicerol, carnitina e betaína tiveram maior concentração na carne de animais alimentados com OS, enquanto que glutamina, glutamato, carnosina e fumarato foram mais presentes no grupo CO. Por outro lado, acetato, succinato, glutamato, colina, glicose, carnosina, acetil-carnosina, glicerato, anserina e inosina monofosfato tiveram maior concentração na carne de animais NE, enquanto que isoleucina, alanina, metionina, creatinina e glicina foram mais presentes na carne de animais NA. Em conclusão, a suplementação com AGPI melhora os índices de desempenho, mas piora as características sensoriais e o índice de qualidade da carne para a saúde humana, independentemente do grupo genético alimentado. Do mesmo modo, a utilização de animais NA melhora os índices de desempenho, mas piora as características sensoriais da carne, enquanto que o grupo genético tem efeitos mínimos sobre o perfil de ácidos graxos da carne e consequentemente não altera a saudabilidade da carne, independentemente da dieta ofertada. Além disso, a partir dos resultados do metaboloma da carne pode-se sugerir que a quantidade de alguns metabólitos, como a carnosina e o glutamato, pode estar relacionada com mudanças na avaliação sensorial do consumidor, principalmente em relação ao sabor da carne. / This work was carried out to evaluate the effect of supplementation with polyunsaturated fatty acids (PUFA) on performance, carcass traits, meat quality attributes and meat metabolome of cattle from different genetic predispositions for intramuscular fat (IMF) deposition. Sixty non-castrated animals, 30 Nellore (NE) and 30 crossbred Nellore x Angus (NA), with an average of 363 ± 28 kg initial body weight and 24 months old, were used in this study. The animals were assigned to a randomized complete block design (initial body weight), with a 2 x 2 factorial arrangement (diet and genetic group), using 15 replicates per treatment. They were evaluated in a 133-d feeding study with one of the following diets: CO - control diet without soybean oil inclusion; SO - control diet containing 3.5% soybean oil inclusion replacing the ground corn grain. Average daily gain (ADG), feed efficiency (G:F), dry matter intake (DMI), and body weight were evaluated. At slaughter, hot carcass weight (HCW), dressing percentage (DP), Longissimus muscle area (LMA), backfat thickness (BFT), and meat marbling score were also evaluated. Samples from Longissimus muscle were used to evaluate the shear force (SC), length of sarcomere, sensorial analysis and meat metabolome were evaluated, as well as shelf life, lipid oxidation, and fatty acid profile and their implication in human health. All data were analyzed using the MIXED procedure of SAS software. There was no diet x genetic group interaction for most traits evaluated, so their effects can be considered separately. In relation to the diet, SO treatment increased ADG (P = 0.0392) and G:F (P = 0.0020) without modifying the DMI and any carcass traits when compared to the CO treatment. There was no effect of the diet in relation to the meat SF and color, but the meat from animals fed SO had lower grades for overall liking (P < 0.0001), tenderness (P < 0.0001) and juiciness (P = 0.0009), besides a trend for lower flavor scores (P = 0.0533) when compared to animals fed CO. No effect of the diet on the amount of IMF was observed, but the meat of animals fed SO diet had a reduction in the proportion of n-3 fatty acids (P = 0.0287), also reducing the n-6: n-3 ratio (P = 0.0256) and the meat quality index (P = 0.0133) for human health. In relation to the genetic group, NA animals had higher IMS (P = 0.0479) and tended to have higher ADG (P = 0.0907) and body weight at slaughter (P = 0.0596) than NE animals, without modifying G:F and any characteristic evaluated in the carcass. No effect of the genetic group on the SF was observed, but the meat from NE animals had higher ratings for overall liking (P = 0.0440) and flavor (P = 0.0444) when compared to the NA animals. In addition, the meat from NE animals had higher values of L* (P = 0.0003) and b* (P = 0.0023) and, consequently, higher values of chroma (P = 0.0358) and hue angle (P = 0.0073) compared to meat from NA animals. There was no effect of the genetic group on the amount of IMF, without any modification in the proportion of fatty acids. Moreover, the meat metabolome was modified (P < 0.10) by both the diet and the genetic group in 8 and 15 metabolites of the 31 identified, respectively. Isoleucine, glycerol, carnitine and betaine had higher concentrations in the meat from animals fed SO, whereas glutamine, glutamate, carnosine and fumarate were in greater concentration in the CO meat. On the other hand, acetate, succinate, glutamate, choline, glucose, carnosine, acetyl-carnosine, glycerate, anserine and inosine monophosphate had higher concentrations in NE meat, whereas isoleucine, alanine, methionine, creatinine and glycine were in greater concentration in the NA meat. In conclusion, PUFA supplementation improves performance rates, but it worsens the sensory characteristics and meat quality index for human health, regardless of the fed genetic group. Similarly, using NA animals improves the performance rates, but it worsens the sensorial characteristics of the meat, whereas the genetic group has minimal effects on the fatty acid profile of the meat and consequently does not alter the healthiness of the meat, regardless of the diet offered. Moreover, from the meat metabolome data, it can be suggested that the amount of some metabolites, such as carnosine and glutamate, may be related to changes in the sensorial evaluation of the consumer, especially in relation to the meat flavor.
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