Prenatal origins of productivity and quality of beef

The productive traits of beef cattle are orchestrated by their genetics, postnatal environmental conditions, and also by the intrauterine background. Both under- or overnutrition, as specific dietary components, are able to promote persistent effects on the offspring. This occurs because dietary factors act not only affecting the availability of substrates for fetal anabolism and oxidative metabolism, but also as signals that regulate several events toward fetal development. Therefore, this study aimed to summarize the gestational nutrition effects on the offspring performance and meat quality in a long term. Overall, studies have shown that many of these alterations are under the control of epigenetic mechanisms, as DNA methylation, histones modification, and non-coding RNA. The current knowledge has indicated that the fetal programming responses are dependent on the window of fetal development in which the dietary treatment is applied, the intensity of maternal nutritional stimuli, and the treatment application length. Collectively, studies demonstrated that muscle cell hyperplasia is impaired when maternal requirements were not achieved in the second third of gestation, which limits the formation of a greater number of muscle fibers and the offspring growth potential in a long term. Changes in muscle fibers metabolism and in collagen content were also reported as consequence of a dietary perturbation during pregnancy. In contrast, a maternal overnutrition during the late pregnancy has been associated with beneficial responses on meat quality. In summary, ensuring an adequate maternal environment during the fetal development is crucial to enhance the productive responses in beef cattle operations.(AU)

Mfsd2a Promotes the Proliferation, Migration, Differentiation and Adipogenesis of Chicken Intramuscular Preadipocytes

Intramuscular fat (IMF) content is a crucial parameter for estimating meat quality. Growing evidence indicates that gene regulation plays an important role in IMF deposition. This study aimed to determine the function of Mfsd2a in chicken intramuscular preadipocytes. In the present study, high Mfsd2a mRNA levels were observed in the liver and adipose tissues of broilers. Subsequently, we synthesized small interfering RNAs to silence the expression of Mfsd2a in chicken intramuscular preadipocytes. The following results suggested that CDK2, PCNA, CCND1, CCND2 and MKI67 were inhibited, with CCK-8 and EdU assays revealing that cell proliferation was inhibited. Scratch test showed that cell migration ratios were declined. We also found that Mfsd2a silencing decreased the mRNA levels of PPARγ, RXRG and their target genes. The similar results were found in some key genes that contribute to lipid synthesis, including C/EBPα, C/EBPß, FABP4, FASN, ACACA and ACSL1. Finally, Oil red O staining showed that IMF accumulation was blocked after Mfsd2a silencing. In conclusion, our results implied that Mfsd2a promotes the proliferation and migration of chicken intramuscular preadipocytes, as well as the differentiation and adipogenesis through PPARγ signaling pathway, which may provide a potential target to improve chicken meat quality.(AU)

Chicken recombinant adiponectin enhances fatty acid metabolism in oleic acid- and palmitic acid-treated LMH cells

The objective of this study was to investigate the effects of recombinant adiponectin on chicken liver cells. The full-length chicken adiponectin gene was amplified by PCR and cloned into the vector pET-32a, followed by the transformation of the vector into Escherichia coli BL21. SDS-PAGE was used to detect and analyze the purity of the expressed recombinant protein. Induction was performed with 1 mM IPTG at 30 °C for 3 h, and the recombinant thioredoxin­adiponectin fusion protein was purified using Ni-NTA affinity chromatography. Chicken adiponectin was successfully expressed and purified in a bacterial system. In addition, the chicken recombinant adiponectin demonstrated that it ameliorates palmitic acid- and oleic acid-induced adipogenesis, in which an increase in ß-oxidation and a decrease in lipogenesis-related genes may be involved. In summary, chicken recombinant adiponectin enhances fatty acid metabolism in LMH cells.(AU)

Potential for in vitro mesoderm differentiation of Wharton's jelly cells from ovine umbilical cord isolated in different culture media / Potencial de diferenciação in vitro de células provenientes da geleia de Wharton de cordões umbilicais ovinos isolados em diferentes meios de cultivo

The mammalian Wharton's jelly of umbilical cord (WJUC) is a promising source of multipotent cells, providing advantages due to ethical implications, ease of collection and the absence of teratomas in pre-clinical trials. Ovine multipotent cells have already been isolated from various tissues, however there are no reports using umbilical cords in this species. This study aimed to investigate the best medium to transport the umbilical cord, to isolate and maintain ovine WJUC cells and to compare in vitro growth and mesodermal differentiation potential. Eight ovine umbilical cords were obtained during parturition, sectioned and transported in six different media: MEM, low glucose DMEM, M199, RPMI 1640, PBS and saline. For each transportation medium, four culture media were used and the tissue was explanted in 24-well plates and cultured in MEM, low glucose DMEM, M199 and RPMI 1640, all with 10% FBS. Every experiment was conducted with low-passage (P2), investigating MTT viability during four days and adipogenic, chondrogenic and osteogenesis differentiation was induced in vitro. The most effective transport medium (p<0.1) was low glucose DMEM. There was no bacterial or fungal contamination from collection. Cells from Wharton's jelly of ovine umbilical cords collected at natural birth possess fibroblastic morphology and the capacity for in vitro differentiation into adipogenic, chondrogenic and osteogenic cell lines. MTT tests and in vitro differentiation experiments revealed that cell culture medium modulates the behavior of cells and is an important factor for proliferation and maintenance of multipotency. Low glucose DMEM was the most suitable medium for the isolation of cells from Wharton's jelly of ovine umbilical cord.(AU)

A geleia de Wharton do cordão umbilical (GWCU) de mamíferos é uma fonte promissora de células multipotentes, sendo vantajosa por aspectos éticos, facilidade de coleta e não causar teratomas em ensaios pré-clínicos. Em ovinos, células multipotentes já foram isoladas de vários tecidos, no entanto, não existem relatos do isolamento a partir do cordão umbilical nesta espécie. O objetivo do presente estudo foi investigar o melhor meio para o transporte do cordão umbilical, isolar e manter as células da GWCU ovino em diferentes meios e comparar a proliferação e o potencial de diferenciação mesodermal in vitro. Oito cordões umbilicais foram obtidos, por ocasião do parto natural, seccionados e transportados em seis diferentes meios que consistiram em MEM, DMEM baixa glicose, M199, RPMI 1640, PBS e soro fisiológico. Para cada meio de transporte foram utilizados quatro meios de cultivo, sendo o tecido explantado em placas de 24 poços e cultivados em MEM, DMEM baixa glicose, M199 e RPMI 1640, todos com 10% SFB. Todo o experimento foi realizado em baixa passagem (P2) investigando viabilidade pelo MTT por quatro dias além da indução à diferenciação adipogênica, condrogênica e osteogênica in vitro. O meio de transporte mais efetivo (P<0,10) foi o DMEM baixa glicose. Não houve contaminações bacterianas ou fúngicas decorrentes da coleta. Células oriundas da geleia de Wharton do cordão umbilical ovino colhido por ocasião do parto natural possuem morfologia fibroblastóide e capacidade de diferenciação in vitro nas linhagens adipogênica, condrogênica e osteogênica. Os ensaios de MTT e diferenciação in vitro, revelaram que o meio de cultura celular modula o comportamento destas células, sendo um fator importante tanto para a proliferação como para a manutenção da multipotência, destacando o DMEM baixa glicose como o meio mais adequado para o transporte e isolamento de células da geleia de Wharton do cordão umbilical ovino.(AU)

Potential for in vitro mesoderm differentiation of Wharton's jelly cells from ovine umbilical cord isolated in different culture media / Potencial de diferenciação in vitro de células provenientes da geleia de Wharton de cordões umbilicais ovinos isolados em diferentes meios de cultivo

The mammalian Wharton's jelly of umbilical cord (WJUC) is a promising source of multipotent cells, providing advantages due to ethical implications, ease of collection and the absence of teratomas in pre-clinical trials. Ovine multipotent cells have already been isolated from various tissues, however there are no reports using umbilical cords in this species. This study aimed to investigate the best medium to transport the umbilical cord, to isolate and maintain ovine WJUC cells and to compare in vitro growth and mesodermal differentiation potential. Eight ovine umbilical cords were obtained during parturition, sectioned and transported in six different media: MEM, low glucose DMEM, M199, RPMI 1640, PBS and saline. For each transportation medium, four culture media were used and the tissue was explanted in 24-well plates and cultured in MEM, low glucose DMEM, M199 and RPMI 1640, all with 10% FBS. Every experiment was conducted with low-passage (P2), investigating MTT viability during four days and adipogenic, chondrogenic and osteogenesis differentiation was induced in vitro. The most effective transport medium (p<0.1) was low glucose DMEM. There was no bacterial or fungal contamination from collection. Cells from Wharton's jelly of ovine umbilical cords collected at natural birth possess fibroblastic morphology and the capacity for in vitro differentiation into adipogenic, chondrogenic and osteogenic cell lines. MTT tests and in vitro differentiation experiments revealed that cell culture medium modulates the behavior of cells and is an important factor for proliferation and maintenance of multipotency. Low glucose DMEM was the most suitable medium for the isolation of cells from Wharton's jelly of ovine umbilical cord.(AU)

A geleia de Wharton do cordão umbilical (GWCU) de mamíferos é uma fonte promissora de células multipotentes, sendo vantajosa por aspectos éticos, facilidade de coleta e não causar teratomas em ensaios pré-clínicos. Em ovinos, células multipotentes já foram isoladas de vários tecidos, no entanto, não existem relatos do isolamento a partir do cordão umbilical nesta espécie. O objetivo do presente estudo foi investigar o melhor meio para o transporte do cordão umbilical, isolar e manter as células da GWCU ovino em diferentes meios e comparar a proliferação e o potencial de diferenciação mesodermal in vitro. Oito cordões umbilicais foram obtidos, por ocasião do parto natural, seccionados e transportados em seis diferentes meios que consistiram em MEM, DMEM baixa glicose, M199, RPMI 1640, PBS e soro fisiológico. Para cada meio de transporte foram utilizados quatro meios de cultivo, sendo o tecido explantado em placas de 24 poços e cultivados em MEM, DMEM baixa glicose, M199 e RPMI 1640, todos com 10% SFB. Todo o experimento foi realizado em baixa passagem (P2) investigando viabilidade pelo MTT por quatro dias além da indução à diferenciação adipogênica, condrogênica e osteogênica in vitro. O meio de transporte mais efetivo (P<0,10) foi o DMEM baixa glicose. Não houve contaminações bacterianas ou fúngicas decorrentes da coleta. Células oriundas da geleia de Wharton do cordão umbilical ovino colhido por ocasião do parto natural possuem morfologia fibroblastóide e capacidade de diferenciação in vitro nas linhagens adipogênica, condrogênica e osteogênica. Os ensaios de MTT e diferenciação in vitro, revelaram que o meio de cultura celular modula o comportamento destas células, sendo um fator importante tanto para a proliferação como para a manutenção da multipotência, destacando o DMEM baixa glicose como o meio mais adequado para o transporte e isolamento de células da geleia de Wharton do cordão umbilical ovino.(AU)

Substâncias obesogênicas alteram a expressão de RNAs longos não-codificadores de maneira transgeracional

A epidemia da obesidade é considerada uma crise mundial de Saúde Pública, tendo como fatores contribuintes o aumento da ingestão calórica, estilos de vida sedentários e/ou predisposições genéticas. Ainda que o balanço energético positivo seja uma das causas mais significativas da obesidade, pesquisas recentes relacionam a epidemia da doença à exposição precoce a produtos químicos desreguladores endócrinos (DEs) (do inglês, Endocrine-Disrupting Chemicals - EDCs). A Sociedade Endócrina define DEs como substâncias químicas exógenas, ou misturas de substâncias químicas, que interferem em qualquer aspecto da ação hormonal. Substâncias obesogênicas compõem um subconjunto de DEs que estimulam inadequadamente a adipogênese e o armazenamento de gordura direta ou indiretamente, aumentando o número e/ou o tamanho das células adiposas, ou interferindo na regulação hormonal do metabolismo, apetite e saciedade. A exposição a essas substâncias químicas durante o período intrauterino e/ou de lactação pode influenciar fortemente a predisposição tardia à obesidade. Os RNAs longos não-codificadores (lncRNAs) são caracterizados por desempenhar relevantes papéis regulatórios em diversos processos biológicos. Considerando a importância da nutrição materna para a regulação dos padrões epigenéticos da prole, o presente estudo visa elucidar os efeitos da exposição ancestral a substâncias obesogênicas na expressão de lncRNAs no tecido adiposo gonadal de camundongos. Os dados de RNA-Seq de 8 amostras de tecido adiposo gonadal de camundongos F4 adultos expostos ancestralmente a dimetilsulfóxido (DMSO) (0,1%) como grupo controle, ou ao DE TBT (50 nM) durante o desenvolvimento uterino e lactação, foram obtidos de estudo previamente realizado (NCBI - GEO Datasets - GSE105051). Após alinhamento e contagem das reads, foi realizada análise de expressão diferencial, e identificamos 74 lncRNAs diferencialmente expressos (p<0.05), visto que 22 apresentaram expressão aumentada no grupo TBT e 52 estavam mais expressos no grupo DMSO em relação ao TBT. As análises dos termos de ontologia gênica (do inglês Gene Ontology - GO) revelaram o enriquecimento dos processos relacionados ao metabolismo da glicose e11 à atividade do receptor ativado por Wnt. Sendo assim, nossos resultados sugerem que a regulação dos lncRNAs e de seus genes parceiros podem contribuir para o controle do processo de adipogênese.

The obesity epidemic is considered a global public health crisis. Increased caloric intake, sedentary lifestyles and/or genetic predisposition are all factors contributing to the epidemic. Although the positive energy balance is one of the most significant causes of obesity, recent research has linked early exposure to Endocrine-Disrupting Chemicals (EDCs) to the disease. The Endocrine Society defines EDCs as "exogenous chemicals, or mixtures of chemicals, that interfere with any aspect of hormonal action". Obesogenic substances make up a subset of EDCs that inadequately stimulate adipogenesis and fat storage directly or indirectly, increasing the number and/or size of fat cells, or interfering with the hormonal regulation of metabolism, appetite and satiety. Exposure to these chemicals during intrauterine and/or lactation periods can strongly influence the late predisposition to obesity. Long noncoding RNAs (lncRNAs) are characterized by playing important regulatory roles in several biological processes. Considering the importance of maternal nutrition to the regulation of epigenetic patterns in the offspring, the present study aimed at investigating the effects of ancestral exposure to obesogenic substances on the expression of lncRNAs in the gonadal adipose tissue of mice. RNA-seq data from 8 gonadal adipose tissue samples from adult F4 mice ancestrally exposed to dimethyl sulfoxide (DMSO) (0.1%) as a control group, or to EDC TBT (50 nM) during uterine development and lactation, were obtained from a previously conducted study (NCBI - GEO Datasets - GSE105051). After reading alignment and counting, differential expression analysis was performed, and we identified 74 differentially expressed lncRNAs were identified (p <0.05), as 22 showed increased expression in the TBT group and 52 were more expressed in the TBT group. The analysis of the GO terms revealed an enrichment for glucose metabolism and the activity of the receptor activated by Wnt. Therefore, our results suggest that the regulation of lncRNAs and their partner genes may contribute to the control of the adipogenesis process.

Development of organs and fat deposition in indigenous goat kids under feed restriction / Desenvolvimento dos órgãos e deposição de gorduras em cabritos Canindé sob restrição alimentar

The study aimed to evaluate the effect of feed restriction on the development of organs and fat deposition in castrated Caninde goats. Twenty one goats were distributed in a completely randomized design, with initial body weight of 15.9 ± 1.03kg. The goats were allocated into three levels of feed restriction (seven animals per level): ad libitum; moderate restriction (restriction of 20% on the amount of feed consumed by the animals fed ad libitum) and severe restriction (40% restriction on the amount of feed consumed by the animals fed ad libitum). The diet consisted of 55% forage (Tifton) and 45% concentrate. At 110 days of experiment, the goats were slaughtered with abody weight of 23.5kg ± 2.5kg. The components included the internal organs (spleen, heart, liver, pancreas, lungs, kidneys, blood) the emptied and cleaned digestive tract and fat deposits (in heart, mesenteric, omental and pelvic-renal) were weighed. Feed restriction in Canindé goats affected the weight of organs and fat deposition in regions more involved with the energy storage function (mesenteric, omental and pelvic-renal) (P 0.05), however, feed restriction does not affect the percentage of the organs in relation to EBW (P > 0.05), indicating that even under feed restriction organ development is proportional to the development of the body.(AU)

Objetivou-se avaliar o efeito da restrição alimentar sobre o desenvolvimento dos órgãos e deposição de gordura em caprinos Canindé castrados. Foram utilizados 21 cabritos confinados, em um delineamento inteiramente casualizado, com peso inicial de 15,9 ± 1,03kg. Os cabritos foram alocados em três níveis de restrição alimentar (sete animais por nível): ad libitum (alimentados à vontade); restrição moderada (restrição de 20% em relação à quantidade de matéria natural consumida pelos animais alimentados ad libitum) e restrição severa (restrição de 40% em relação à quantidade de ração consumida pelos animais alimentados ad libitum). A ração experimental apresentou uma proporção de 55% de volumoso (Tifton) e 45% de concentrado. Aos 110 dias de experimento os cabritos foram abatidos com peso médio de 23,5 kg ± 2,5 kg. Esvaziou-se o trato gastrointestinal (TGI), a bexiga e a vesícula biliar e foram mensurados os seus pesos para determinação do peso de corpo vazio (PCV). Foram separados e registrados os pesos dos órgãos (baço, coração, fígado, pâncreas, pulmões, rins, sangue, TGI) e dos depósitos de gordura (cardíaca, mesentérica, omental e pélvico-renal). A restrição alimentar em caprinos Canindé afetou o peso absoluto dos órgãos e das gorduras (P 0.05) que estão mais envolvidas com a função de reserva energética (mesentérica, omental e pélvico-renal), no entanto, não afetou o pe(AU)

Análise da expressão de genes associados à adipogênese e composição da gordura intramuscular em bovinos Angus X Nelore de diferentes condições sexuais

A condição sexual dos bovinos influencia o seu crescimento, assim como a formação do tecido adiposo muscular, a composição de ácidos graxos em diferentes músculos e vias de adipogênese, sendo estas influenciadas por uma variedade de sinais hormonais, o que afeta diretamente as características da carcaça e a qualidade da carne. Desta forma, o presente estudo analisou a influência da condição sexual (CS) dos bovinos sobre a expressão dos genes associados à adipogênese, assim como avaliou o perfil de ácidos graxos dos músculos longissimus (LO) e triceps brachii (TB) de bovinos Angus x Nelore. Foram confinados 150 bovinos, divididos em três CS (50 fêmeas, 50 machos não castrados e 50 castrados). O delineamento experimental considerado foi inteiramente casualizado, com 50 repetições por grupo e cada animal foi considerado como uma unidade experimental. Os animais foram mantidos nas mesmas condições de manejo, recebendo alimentação ad libitum utilizando-se a mesma dieta, composta de 86% de concentrado e 14% de volumoso na matéria seca. Após 150 dias de confinamento, os animais foram abatidos aos 16 meses de idade. Durante o abate, imediatamente após a esfola foram coletadas amostras dos músculos LO e TB para extração de RNA e proteína. Durante a desossa foi avaliada a espessura de gordura subcutânea e o escore de marmorização no músculo longissimus, entre a 12ª e 13ª costelas e coletadas amostras dos dois músculos para análise de quantificação de lipídeos totais e composição de ácidos graxos. Os dados estatísticos referentes à qualidade de carcaça e carne foram analisados pelo proc MIXED do programa SAS (versão 9.3) e os referentes aos resultados de expressão gênica pelo programa JMP14Ò (SAS Institute). A CS afetou as características de carcaça avaliadas (P 0,05). A espessura de gordura subcutânea e o escore de marmorização foram maiores na carcaça das fêmeas em comparação às dos machos não castrados e castrados. Houve diferença no conteúdo de lipídeos totais somente entre as CS (P 0,05), com maior conteúdo na carne das fêmeas, seguida da carne dos machos castrados e não castrados. O total de ácidos graxos saturados (AGS) foi influenciado pela CS e pelo tipo de músculo (P 0,05), sendo que a carne dos bovinos não castrados apresentou maior concentração de AGS do que a carne das fêmeas. O total de AGS no músculo LO foi maior que o TB, independente da CS. Já os ácidos graxos (AG) 14:0 e 16:0 foram afetados apenas pelo tipo de músculo, e concentrações mais elevadas foram detectadas no músculo LO (P 0,05). O fator CS afetou o total de AG monoinsaturados (AGMI) (P 0,05), sendo a maior concentração observada na carne de fêmeas e castrados do que na carne de não castrados. Já a concentração do 18:1 n-9c (oleico) foi influenciada pelo tipo de músculo, com maior concentração no músculo LO em comparação ao TB (P 0,05). Houve efeito do tipo de músculo para o total de AG poli-insaturados (AGPI), total de n-3, total de n-6, proporção AGPI:AGS, os AG individuais 18:2 n-6c, 20:3 n-3, 22:4 n-6 e o índice de saúde. As concentrações foram maiores no TB em comparação ao LO (P 0,05). Por outro lado, os índices de aterogenicidade e trombogenicidade foram maiores no músculo LO (P 0,05). Com relação aos níveis de transcritos, houve efeito do tipo de músculo na expressão dos genes CEBPa e LPL (P 0,05), ambos com maior expressão no músculo TB. Já a CS influenciou o nível de transcritos do ACC (P 0,05) e o gene foi mais expresso nas fêmeas em relação aos machos castrados. Foi observada interação entre CS e tipo de músculo para os genes FABP3, TPM2 e TPM3 (P 0,05). Houve diferença na abundância da proteína tropomiosina (TPM) entre os músculos, mas somente dentro da condição sexual machos não castrados (P £ 0,05), com maior abundância no músculo TB em comparação ao LO. Adicionalmente, a CSafetou a abundância da TPM avaliada nos músculos LO e TB (P £ 0,05). O músculo LO dos machos castrados apresentou maior abundância da TPM do que os machos não castrados e fêmeas, enquanto o TB dos machos não castrados apresentou maior abundância do que as demais CS. A CS modulou os níveis de transcritos do ACC, conteúdo de lipídeos totais, total de AGS, total de AGMI e as características de carcaça, enquanto o tipo de músculo afetou os transcritos dos genes CEBPa e LPL, total de AGS, total de AGPI, total de n-3 e n-6, além de importantes AG, demonstrando efeito especifico da CS e o tipo de músculo. Adicionalmente, foi observada uma modulação da tropomiosina na formação e acúmulo de lipídeos, relacionada a maior espessura de gordura subcutânea e escore de marmorização nas fêmeas e machos castrados, e uma modulação desta mesma proteína no desenvolvimento muscular, apontada pela menor deposição de gordura na carcaça de machos não castrados. Além disso, há um perfil lipídico diferente dentro de cada músculo, onde o TB é mais favorável à saúde humana do que o LO, e a carne bovina de fêmeas se destaca das demais condições sexuais por apresentar melhores características de carcaça e composição de AG mais favorável à saúde humana.

Cattle growth, muscle adipose tissue and fatty acid composition in different muscles, are significantly affected by gender and influenced by a variety of hormonal factors, affecting the quality of the carcass and the beef. As the adipogenesis is a complex biological process to regulate fat cell metabolism, it can be regulated by hormones and influenced by factors such as different fat deposits, the gender status and diet of cattle. Therefore, it is important to understand the molecular mechanisms involved in adipogenesis in different muscles and gender, since the deposition of fat in these animals is different, to employ new tools to add greater value to meat production. However, the present study evaluated the association of fatty acid (FA) composition and adipogenesis-related genes in the longissimus (LO) and triceps brachii (TB) muscles of feedlot Angus x Nelore cattle. A total of 150 cattle was confined for 150 days and assigned to three genders, namely, heifers, bulls, and steers, and fed the same diet. Immediately after slaughter, samples of LO and TB muscles were collected for RNA and protein extraction. Twenty-four hours after slaughter, were measured the fat thickness and marbling score between the 12th and 13th ribs and collected samples of the muscles for analyses of total lipids determination and fatty acid composition. The experimental design was completely randomized, with 50 repetitions per treatment, and each animal was considered an experimental unit. The statistical analyses were performed by the proc MIXED SAS® (version 9.3) for the meat quality traits and JMP14 Software (SAS Institute) for the gene expression analyses. There was difference in the carcass traits related to fat deposition (P £ 0.05). The fat thickness and marbling score were higher in heifer carcasses than in carcasses of bulls and steers. Similarly, only gender status affected the content of total lipids (P £ 0.05), where meat from heifers presented increased values compared to steers and bulls. There was a difference in the total saturated FAs (SFAs) by gender and muscle type (P £ 0.05). The meat from bulls had higher levels of SFAs compared to heifers and had no differences in relation to meat from steers. Among muscle types, total SFAs increased in LO muscle compared with TB, independent of gender. The FAs 14:0 and 16:0 were affected by muscle type only, and higher levels were detected in the LO muscle (P £ 0.05). There were differences in gender for total monounsaturated FAs (MUFAs; P £ 0.05). Meat from bulls had lower levels of MUFAs compared with the other genders. The levels of the major FA of total MUFAs, 18:1 n-9c, were higher in the LO muscle than in the TB muscle, independent of gender (P £ 0.05). The total polyunsaturated FAs (PUFAs), total n-3, total n-6, PUFA:SFA ratio, health index and FAs 18:2 n-6c, 20:3 n-3, and 22:4 n-6 were higher in TB than in LO muscle (P < 0.05). Otherwise, the atherogenicity and thrombogenicity indexes were higher in the LO muscle (P £ 0.05). Regarding the transcript levels, there was an effect of muscle type on the expression of the CEBPa and LPL (P £ 0.05) genes, both of which had higher expression in TB muscle. Gender affected the transcript level of ACC (P £ 0.05). This gene was increased in bulls, intermediate in steers, and decreased in heifers. Additionally, gender and muscle type interactions were observed for the transcript levels of FABP3, TPM2, and TPM3 (P £ 0.05). There was difference for tropomyosin (TPM) protein abundance between muscles only in bulls (P £ 0.05), where the greater TPM abundance was in the TB. Additionally, cattle gender status affected the TPM abundance evaluated in LO and TB muscles (P 0.05). The LO muscle of steers showed higher TPM abundance than bulls and heifers, while the TB muscle of bulls showed higher TPMabundance than the other genders. The cattle gender status modulated the transcripts levels of ACC gene, content of total lipids, total SFAs, total MUFAs, and carcass traits, while the muscle type affected the transcripts of CEBPa and LPL genes, total SFAs, total PUFAs, total n-3 and n-6, besides important FAs, showing a gender and muscle specific effect. In addition, a modulation of TPM in formation and accumulation of lipids was observed, related to the greater subcutaneous fat thickness and marbling score in heifers and steers, and a modulation of this same protein in muscle development, indicated by the lower fat deposition in the carcass of bulls. Furthermore, there is a different lipid profile within each muscle, where the TB muscle is more favorable to human health than the LO, and beef from heifers stands out than the other genders for presenting better carcass traits and FA composition more favorable to human health.

Development of organs and fat deposition in indigenous goat kids under feed restriction / Desenvolvimento dos órgãos e deposição de gorduras em cabritos Canindé sob restrição alimentar

The study aimed to evaluate the effect of feed restriction on the development of organs and fat deposition in castrated Caninde goats. Twenty one goats were distributed in a completely randomized design, with initial body weight of 15.9 ± 1.03kg. The goats were allocated into three levels of feed restriction (seven animals per level): ad libitum; moderate restriction (restriction of 20% on the amount of feed consumed by the animals fed ad libitum) and severe restriction (40% restriction on the amount of feed consumed by the animals fed ad libitum). The diet consisted of 55% forage (Tifton) and 45% concentrate. At 110 days of experiment, the goats were slaughtered with abody weight of 23.5kg ± 2.5kg. The components included the internal organs (spleen, heart, liver, pancreas, lungs, kidneys, blood) the emptied and cleaned digestive tract and fat deposits (in heart, mesenteric, omental and pelvic-renal) were weighed. Feed restriction in Canindé goats affected the weight of organs and fat deposition in regions more involved with the energy storage function (mesenteric, omental and pelvic-renal) (P 0.05), however, feed restriction does not affect the percentage of the organs in relation to EBW (P > 0.05), indicating that even under feed restriction organ development is proportional to the development of the body.

Objetivou-se avaliar o efeito da restrição alimentar sobre o desenvolvimento dos órgãos e deposição de gordura em caprinos Canindé castrados. Foram utilizados 21 cabritos confinados, em um delineamento inteiramente casualizado, com peso inicial de 15,9 ± 1,03kg. Os cabritos foram alocados em três níveis de restrição alimentar (sete animais por nível): ad libitum (alimentados à vontade); restrição moderada (restrição de 20% em relação à quantidade de matéria natural consumida pelos animais alimentados ad libitum) e restrição severa (restrição de 40% em relação à quantidade de ração consumida pelos animais alimentados ad libitum). A ração experimental apresentou uma proporção de 55% de volumoso (Tifton) e 45% de concentrado. Aos 110 dias de experimento os cabritos foram abatidos com peso médio de 23,5 kg ± 2,5 kg. Esvaziou-se o trato gastrointestinal (TGI), a bexiga e a vesícula biliar e foram mensurados os seus pesos para determinação do peso de corpo vazio (PCV). Foram separados e registrados os pesos dos órgãos (baço, coração, fígado, pâncreas, pulmões, rins, sangue, TGI) e dos depósitos de gordura (cardíaca, mesentérica, omental e pélvico-renal). A restrição alimentar em caprinos Canindé afetou o peso absoluto dos órgãos e das gorduras (P 0.05) que estão mais envolvidas com a função de reserva energética (mesentérica, omental e pélvico-renal), no entanto, não afetou o pe

Desenvolvimento dos órgãos e deposição de gorduras em cabritos Canindé sob restrição alimentar

SUMMARY The study aimed to evaluate the effect of feed restriction on the development of organs and fat deposition in castrated Caninde goats. Twenty one goats were distributed in a completely randomized design, with initial body weight of 15.9 ± 1.03kg. The goats were allocated into three levels of feed restriction (seven animals per level): ad libitum; moderate restriction (restriction of 20% on the amount of feed consumed by the animals fed ad libitum) and severe restriction (40% restriction on the amount of feed consumed by the animals fed ad libitum). The diet consisted of 55% forage (Tifton) and 45% concentrate. At 110 days of experiment, the goats were slaughtered with abody weight of 23.5kg ± 2.5kg. The components included the internal organs (spleen, heart, liver, pancreas, lungs, kidneys, blood) the emptied and cleaned digestive tract and fat deposits (in heart, mesenteric, omental and pelvic-renal) were weighed. Feed restriction in Canindé goats affected the weight of organs and fat deposition in regions more involved with the energy storage function (mesenteric, omental and pelvic-renal) (P 0.05), however, feed restriction does not affect the percentage of the organs in relation to EBW (P > 0.05), indicating that even under feed restriction organ development is proportional to the development of the body.

RESUMO Objetivou-se avaliar o efeito da restrição alimentar sobre o desenvolvimento dos órgãos e deposição de gordura em caprinos Canindé castrados. Foram utilizados 21 cabritos confinados, em um delineamento inteiramente casualizado, com peso inicial de 15,9 ± 1,03kg. Os cabritos foram alocados em três níveis de restrição alimentar (sete animais por nível): ad libitum (alimentados à vontade); restrição moderada (restrição de 20% em relação à quantidade de matéria natural consumida pelos animais alimentados ad libitum) e restrição severa (restrição de 40% em relação à quantidade de ração consumida pelos animais alimentados ad libitum). A ração experimental apresentou uma proporção de 55% de volumoso (Tifton) e 45% de concentrado. Aos 110 dias de experimento os cabritos foram abatidos com peso médio de 23,5 kg ± 2,5 kg. Esvaziou-se o trato gastrointestinal (TGI), a bexiga e a vesícula biliar e foram mensurados os seus pesos para determinação do peso de corpo vazio (PCV). Foram separados e registrados os pesos dos órgãos (baço, coração, fígado, pâncreas, pulmões, rins, sangue, TGI) e dos depósitos de gordura (cardíaca, mesentérica, omental e pélvico-renal). A restrição alimentar em caprinos Canindé afetou o peso absoluto dos órgãos e das gorduras (P 0.05) que estão mais envolvidas com a função de reserva energética (mesentérica, omental e pélvico-renal), no entanto, não afetou o percentual dos órgãos em relação ao PCV (P > 0.05), indicando que mesmo sob restrição o desenvolvimento dos órgãos é proporcional ao desenvolvimento do corpo.

Desenvolvimento dos órgãos e deposição de gorduras em cabritos Canindé sob restrição alimentar

SUMMARY The study aimed to evaluate the effect of feed restriction on the development of organs and fat deposition in castrated Caninde goats. Twenty one goats were distributed in a completely randomized design, with initial body weight of 15.9 ± 1.03kg. The goats were allocated into three levels of feed restriction (seven animals per level): ad libitum; moderate restriction (restriction of 20% on the amount of feed consumed by the animals fed ad libitum) and severe restriction (40% restriction on the amount of feed consumed by the animals fed ad libitum). The diet consisted of 55% forage (Tifton) and 45% concentrate. At 110 days of experiment, the goats were slaughtered with abody weight of 23.5kg ± 2.5kg. The components included the internal organs (spleen, heart, liver, pancreas, lungs, kidneys, blood) the emptied and cleaned digestive tract and fat deposits (in heart, mesenteric, omental and pelvic-renal) were weighed. Feed restriction in Canindé goats affected the weight of organs and fat deposition in regions more involved with the energy storage function (mesenteric, omental and pelvic-renal) (P 0.05), however, feed restriction does not affect the percentage of the organs in relation to EBW (P > 0.05), indicating that even under feed restriction organ development is proportional to the development of the body.

RESUMO Objetivou-se avaliar o efeito da restrição alimentar sobre o desenvolvimento dos órgãos e deposição de gordura em caprinos Canindé castrados. Foram utilizados 21 cabritos confinados, em um delineamento inteiramente casualizado, com peso inicial de 15,9 ± 1,03kg. Os cabritos foram alocados em três níveis de restrição alimentar (sete animais por nível): ad libitum (alimentados à vontade); restrição moderada (restrição de 20% em relação à quantidade de matéria natural consumida pelos animais alimentados ad libitum) e restrição severa (restrição de 40% em relação à quantidade de ração consumida pelos animais alimentados ad libitum). A ração experimental apresentou uma proporção de 55% de volumoso (Tifton) e 45% de concentrado. Aos 110 dias de experimento os cabritos foram abatidos com peso médio de 23,5 kg ± 2,5 kg. Esvaziou-se o trato gastrointestinal (TGI), a bexiga e a vesícula biliar e foram mensurados os seus pesos para determinação do peso de corpo vazio (PCV). Foram separados e registrados os pesos dos órgãos (baço, coração, fígado, pâncreas, pulmões, rins, sangue, TGI) e dos depósitos de gordura (cardíaca, mesentérica, omental e pélvico-renal). A restrição alimentar em caprinos Canindé afetou o peso absoluto dos órgãos e das gorduras (P 0.05) que estão mais envolvidas com a função de reserva energética (mesentérica, omental e pélvico-renal), no entanto, não afetou o percentual dos órgãos em relação ao PCV (P > 0.05), indicando que mesmo sob restrição o desenvolvimento dos órgãos é proporcional ao desenvolvimento do corpo.

Isolation, expansion, differentiation and growth kinetics essay in mesenchymal stem cells culture from the bone marrow of collared peccaries (Tayassu tajacu)

Background: There are few studies on stem cell isolation in wild animals that provide isolation and culture protocols of these cells in vitro. Among the wild species studied, we present the collared peccary (Tayassu tajacu) as a model with potential to obtain and use MSC in preclinical studies. These animals are phylogenetically close to the domestic pig, popularly known as peccaries and found naturally in South America, Central America and the South of the United States. The aim of the present study was to establish a protocol for the isolation, in vitro cell expansion, differentiation and assessment of the stromal MSC growth curve before and after thawing. Materials, Methods & Results: Mesenchymal stem cells (MSC) from collared peccary bone marrow (Tayassu tajacu) were isolated and expanded by centrifuge in Ficoll® solution and cultured in DMEM® High Glucose medium. The culture was assessed by assays of colony forming units CFU-F and growth curve by saturation (GCS). Cultures in the third passage, with 70% confluence, were replicated at 105 cells/mL concentration in the culture media to induce osteogenic cell differentiation and adipogenic cell differentiation, respectively. The MSC were frozen in nitrogen for 40 days, thawed and re-assessed for cell viability and GCS. Discussion: The bone marrow collected presented high mononuclear cellularity, with a mean variability of 94.5% and 60.83 ± 4.27 UFC were identified in the samples and cells with fibroblast-like-cell morphology were observed. When they were expanded, the mean cell viability was 95%, the mean cell concentration obtained was 233.31 ± 20.04 cells per 25cm2 bottle and the culture reached the growth plateau in GCS between the 13th and 16th day. The osteoblastic cell differentiation assay showed after 18 days, morphology similar to osteoblasts, with irregular cytoplasm limits, cell prolongation formation and flattened appearance. [...](AU)

Isolation, expansion, differentiation and growth kinetics essay in mesenchymal stem cells culture from the bone marrow of collared peccaries (Tayassu tajacu)

Background: There are few studies on stem cell isolation in wild animals that provide isolation and culture protocols of these cells in vitro. Among the wild species studied, we present the collared peccary (Tayassu tajacu) as a model with potential to obtain and use MSC in preclinical studies. These animals are phylogenetically close to the domestic pig, popularly known as peccaries and found naturally in South America, Central America and the South of the United States. The aim of the present study was to establish a protocol for the isolation, in vitro cell expansion, differentiation and assessment of the stromal MSC growth curve before and after thawing. Materials, Methods & Results: Mesenchymal stem cells (MSC) from collared peccary bone marrow (Tayassu tajacu) were isolated and expanded by centrifuge in Ficoll® solution and cultured in DMEM® High Glucose medium. The culture was assessed by assays of colony forming units CFU-F and growth curve by saturation (GCS). Cultures in the third passage, with 70% confluence, were replicated at 105 cells/mL concentration in the culture media to induce osteogenic cell differentiation and adipogenic cell differentiation, respectively. The MSC were frozen in nitrogen for 40 days, thawed and re-assessed for cell viability and GCS. Discussion: The bone marrow collected presented high mononuclear cellularity, with a mean variability of 94.5% and 60.83 ± 4.27 UFC were identified in the samples and cells with fibroblast-like-cell morphology were observed. When they were expanded, the mean cell viability was 95%, the mean cell concentration obtained was 233.31 ± 20.04 cells per 25cm2 bottle and the culture reached the growth plateau in GCS between the 13th and 16th day. The osteoblastic cell differentiation assay showed after 18 days, morphology similar to osteoblasts, with irregular cytoplasm limits, cell prolongation formation and flattened appearance. [...]

Betaine and ractopamine for pigs from 90 to 140 kg

Foram conduzidos dois experimentos com fêmeas suínas dos 90 aos 140 kg. No primeiro estudo (I), o objetivo foi avaliar a suplementação de ractopamina e betaína, isolada ou em associação, no desempenho, características de carcaça, qualidade de carne e na expressão de genes relacionados ao metabolismo lipídico no tecido muscular glicolítico e oxidativo e no tecido adiposo subcutâneo. No segundo experimento (II), objetivou-se avaliar o desempenho, características de carcaça, qualidade de carne e a expressão de genes relacionados ao metabolismo lipídico no tecido muscular glicolítico e oxidativo de duas linhagens genéticas distintas consumindo ractopamina. No primeiro experimento, foram utilizadas 72 fêmeas híbridas (Agroceres PIC), com peso médio inicial de 88,96 ± 3,44 kg, distribuídos em delineamento inteiramente casualizado, com quatro tratamentos, nove repetições e dois animais por unidade experimental. Os tratamentos foram constituídos por uma ração controle (CONT), CONT + 2,5 g/kg de betaína (BET), ração com 20 ppm de ractopamina (RAC), RAC + 2,5 g/kg de betaína (RAC + BET). No segundo experimento, foram utilizadas 18 fêmeas Agroceres PIC 337 (Large White x Landrace x Duroc x Pietrain) x Camborough (Large White x Landrace) (HIB), com peso médio inicial de 88,96± 3,44 kg e 12 fêmeas DB LQ 1250 (Duroc) x DB 90 (Large White x Landrace) (DUR), com peso médio inicial de 85,63± 1,55 kg, distribuídos em delineamento inteiramente casualizado com dois tratamentos (Híbrido) ou (Duroc), com 9 e 6 repetições, respectivamente, e dois animais por unidade experimental. No experimento I, suínos consumindo RAC e RAC+BET apresentaram maior ganho de peso diário, menor conversão alimentar comparado às demais dietas e maior peso de carcaça quente e rendimento de carcaça em relação à dieta CONT. Suínos que consumiram as dietas BET, RAC e RAC+BET apresentaram maior área de olho de lombo, enquanto a espessura de toucinho foi menor nos animais que receberam a dieta RAC+BET em comparação ao CONT. A carne de animais que receberam a dieta BET apresentou menor força de cisalhamento e maior percentual de gordura intramuscular comparado ao CONT. A BET aumentou a expressão de fatores lipogênicos no Longissimus dorsi e no músculo Sóleo em relação ao CONT. Em relação ao tecido adiposo, RAC e BET aumentaram a expressão de genes relacionados a lipólise e -oxidação. Os resultados indicam que o desempenho e as características de carcaça podem ser aumentados com o uso da ractopamina, enquanto a betaína promove aumento na área de olho de lombo e na qualidade da carne suína. No experimento II, o desempenho dos suínos de ambos os grupos genéticos foi semelhante. No entanto, os suínos DUR apresentaram menor espessura de toucinho. A carne dos suínos DUR apresentou maior L* (luminosidade), menor a* (intensidade de vermelho) e maior percentual de gordura intramuscular. Suínos DUR apresentaram maior expressão do receptor ativado por proliferadores de peroxissoma gama (PPAR) e da proteína ácido graxo translocase/cluster de diferenciação (FAT/CD36), além de maior abundância do mRNA da lipase hormônio sensível (HSL) e Adiponectina (ADIP) no Longissimus Dorsi. Maior abundância do mRNA do PPAR e FAT/CD36 também foi observado no músculo Sóleo dos animais DUR, mas sem alteração na expressão de enzimas relacionadas à degradação lipídica comparado a suínos HIB. Suínos DUR apresentam carcaça com menor espessura de toucinho e melhor qualidade de carne, evidenciado pelo maior percentual de gordura intramuscular em relação aos suínos HIB. Possivelmente a maior concentração de gordura intramuscular seja devido principalmente a maior expressão de fatores de transcrição e enzimas relacionados a adipogêneses e lipogêneses no tecido muscular esquelético glicolítico e oxidativo nos suínos DUR.

Two experiments were carried out with female pigs from 90 to 140 kg. In experiment I, the objective was to evaluate the supplementation of ractopamine and betaine, alone or in association, on performance, carcass traits, meat quality and expression of genes related to lipid metabolism in skeletal glycolytic and oxidative muscle tissue and in the subcutaneous adipose tissue. In experiment II, to evaluate performance, carcass traits, pork quality and expression of genes related to lipid metabolism in glycolytic and oxidative muscle tissues of two divergent genetic lines fed ractopamine. In experiment I, a total of 72 female pigs (Agroceres PIC) with initial body weight of 88.96 ± 3.44 kg were assigned to a completely randomized design in four diets with nine replicates and two pigs per experimental unit, represented by the pen. Diets consisted of a control (CTRL) diet (without ractopamine and betaine); CTRL + 2.5 g/kg betaine (BET); CTRL+ 20 ppm ractopamine (RAC); and RAC + 2.5 g/kg de betaine (RAC + BET). In experiment II, eighteen female pigs Agroceres PIC 337 (Large White x Landrace x Duroc x Pietrain) x Camborough (Large White x Landrace) (HYB), with initial body weight of 88.96 ± 3.44 kg and 12 female pigs DB LQ 1250 (Duroc) x DB 90 (Large White x Landrace) (DUR), with initial body weight of 85.63 ± 1.55 kg were used. Pigs were allotted in a completely randomized design in two groups: HYB and DUR with nine and six replicates, respectively, and two pigs per experimental unit, represented by the pen. In experiment I, pigs fed RAC and RAC+BET diets had higher daily weight gain, lower feed conversion compared to pigs fed the other diets and higher hot carcass weight and carcass yield, compared to CTRL diet. Pigs fed BET, RAC and RAC+BET diets had greater loin muscle area. The backfat thickness was lower in pigs fed RAC+BET diet, compared to the CTRL. The meat from pigs fed BET diet had lower Warner-Bratzler shear force and higher percentage of intramuscular fat compared to CTRL. BET diet increased the expression of lipogenic factors in Longissimus dorsi and Soleus muscles compared to CTRL. In adipose tissue, RAC and BET diets increased the expression of genes related to lipolysis and -oxidation. The results indicate that performance and carcass traits can be improved with ractopamine and betaine promotes increase in loin muscle area and pork quality. In experiment II, the performance of pigs from both genetic groups was similar. However, DUR pigs had lower backfat thickness. The meat from DUR pigs had higher L* (brightness), lower a* (redness) and higher percentage of intramuscular fat. DUR pigs had higher expression of peroxisome proliferator-activated receptor gamma (PPAR) and fatty acid protein translocase/cluster differentiation (FAT/CD36), in addition to higher abundance of hormone-sensitive lipase (HSL) and Adiponectin (ADIP) mRNA in the Longissimus dorsi. Greater abundance of PPAR and FAT/CD36 mRNA was also observed in the Soleus muscle of DUR pigs with no change in the expression of enzymes related to lipid degradation compared to HYB pigs. DUR pigs had carcasses with lower backfat thickness and better meat quality, evidenced by the higher percentage of intramuscular fat compared to HYB pigs. Possibly the higher concentration of intramuscular fat is mainly due to the higher expression of transcription factors and enzymes related to adipogenesis and lipogenesis in glycolytic and oxidative skeletal muscle tissue in DUR pigs

Isolation, expansion and differentiation of cellular progenitors obtained from dental pulp of agouti (Dasyprocta prymnolopha Wagler, 1831) / Isolamento, expansão e diferenciação de progenitors celulares obtidos da polpa dentária de cutia (Dasyprocta prymnolopha Wagler, 1831)

The study aimed to isolate, expand, differentiate and characterize progenitor cells existent in the dental pulp of agouti. The material was washed with PBS solution and dissociated mechanically with the aid of a scalpel blade on plates containing culture medium D-MEM/F-12, and incubated at 5% CO2-37⁰C. The growth curve, CFU assay, osteogenic/adipogenic differentiation and characterization were obtained from the isolation. The cells began to be released from the explant tissue around the 7th day of culture. By day 22 of culture, cells reached 80% confluence. At the UFC test, 81 colonies were counted with 12 days of cultivation. The growth curves before and after freezing showed a regular growth with intense proliferation and clonogenic potential. The cell differentiation showed formation of osteoblasts and fat in culture, starting at 15 days of culture in a specific medium. Flow cytometry (FACs) was as follows: CD34 (positive), CD14 (negative), CD45 (negative), CD73 (positive), CD79 (negative), CD90 (positive), CD105 (positive), demonstrating high specificity and commitment of isolated cells with mesenchymal stem cells strains. These results suggest the existence of a cell population of stem cells with mesenchymal features from the isolated tissue in the explants of agouti dental pulp, a potential model for study of stem cell strains obtained from the pulp tissue.(AU)

Isolation, expansion and differentiation of cellular progenitors obtained from dental pulp of agouti (Dasyprocta prymnolopha Wagler, 1831). Este estudo teve como objetivo isolar, expandir, diferenciar e caracterizar células progenitoras existentes na polpa dentária de cutia. O material foi lavado em solução de PBS e dissociado mecanicamente, com o auxílio de uma lâmina de bisturi, em placas contendo meio de cultura D-MEM/F-12, e incubadas em 5% de CO2-37⁰C. A curva de crescimento, o ensaio de CFU, a diferenciação osteogênica/adipogênica e a caracterização foram obtidas a partir do isolamento. As células começaram a ser liberadas, a partir do explante, em torno do sétimo dia de cultura. A partir do 22o dia, as células atingiram 80% de confluência. No teste para UFC, 81 colônias foram contadas aos 12 dias de cultivo. As curvas de crescimento pré- e pós-congelamento apresentaram crescimento regular, com intensa proliferação e potencial clonogênico. A diferenciação das células mostrou a formação de osteoblastos e de células de gordura, a partir de 15 dias de cultura em meio específico. A citometria de fluxo (FACS) apresentou-se como segue: CD34 (positivo), CD14 (negativo), CD45 (negativo), CD73 (positivo), CD79 (negativo), CD90 (positivo), CD105 (positivo), demonstrando a grande especificidade e comprometimento das células isoladas com linhagens de células-tronco mesenquimais. Estes resultados sugerem a existência de uma população de células-tronco mesenquimais isolada a partir de explantes da polpa dentária cutia, um modelo potencial para o estudo de linhagens de células-tronco obtidas a partir do tecido pulpar.(AU)

The Expression of Can and Camk is Associated with Lipogenesis in the Muscle of Chicken

Intramuscular fat (IMF) content in chickens significantly contributes to meat quality. The main objective of this study was to assess the expression of calcineurin (CaN) and Ca2+/calmodulin-dependent protein kinase (CaMK) in lipogenesis in chicken muscle. Chickens were slaughtered and sampled at 4, 8, and 16 weeks of age. IMF content and the expression of CaN subunits and CaMK isoforms were measured in the thigh muscle tissue. The results showed that the IMF contents were greater at 16 weeks compared with those at 4 and 8 weeks (p 0.05). Transcription of fatty acid synthase (FAS) and fatty acid translocase CD36 (FAT/CD36) mRNA significantly increased with age, from four to 16 weeks (p 0.05). The mRNA levels of CaNB and CaMK IV were significantly lower at 16 weeks than at four weeks (p 0.05), but CaMK II mRNA levels were significantly higher than at four weeks (p 0.05). In order to evaluate the role of CaMK and CaN in adipogenesis, SV cells were incubated in standard adipogenic medium for 24 h and treated with specific inhibitor of CaMK and CaN. The expressions of CCAAT/enhancer binding protein b (C/EBPb, sterol regulatory element-binding protein 1 (SREBP1),and peroxisome proliferation-activated receptor g (PPAR)were dramatically enhanced by the CsA, CaN inhibitor (p 0.05). KN93, CaMK II inhibitor, dramatically repressed the expression of those lipogenic gene (p 0.05). These results indicated that CaN and CaMK had different effects on adipogenesis in the muscle of chickens.(AU)

The Expression of Can and Camk is Associated with Lipogenesis in the Muscle of Chicken

Intramuscular fat (IMF) content in chickens significantly contributes to meat quality. The main objective of this study was to assess the expression of calcineurin (CaN) and Ca2+/calmodulin-dependent protein kinase (CaMK) in lipogenesis in chicken muscle. Chickens were slaughtered and sampled at 4, 8, and 16 weeks of age. IMF content and the expression of CaN subunits and CaMK isoforms were measured in the thigh muscle tissue. The results showed that the IMF contents were greater at 16 weeks compared with those at 4 and 8 weeks (p 0.05). Transcription of fatty acid synthase (FAS) and fatty acid translocase CD36 (FAT/CD36) mRNA significantly increased with age, from four to 16 weeks (p 0.05). The mRNA levels of CaNB and CaMK IV were significantly lower at 16 weeks than at four weeks (p 0.05), but CaMK II mRNA levels were significantly higher than at four weeks (p 0.05). In order to evaluate the role of CaMK and CaN in adipogenesis, SV cells were incubated in standard adipogenic medium for 24 h and treated with specific inhibitor of CaMK and CaN. The expressions of CCAAT/enhancer binding protein b (C/EBPb, sterol regulatory element-binding protein 1 (SREBP1),and peroxisome proliferation-activated receptor g (PPAR)were dramatically enhanced by the CsA, CaN inhibitor (p 0.05). KN93, CaMK II inhibitor, dramatically repressed the expression of those lipogenic gene (p 0.05). These results indicated that CaN and CaMK had different effects on adipogenesis in the muscle of chickens.

EFEITO DO MÉRITO GENÉTICO PARA ACABAMENTO DE CARCAÇA E DA RAÇA PATERNA SOBRE O DESEMPENHO, EXPRESSÃO GÊNICA E QUALIDADE DA CARNE DE NOVILHOS DE CORTE

A deposição de gordura subcutânea em novilhos de corte é de grande relevância e o valor genético dos reprodutores pode contribuir para o entendimento e controle eficiente desta característica nos sistemas produtivos de carne bovina. Objetivou-se avaliar os efeitos do mérito genético para acabamento de carcaça (MGA) sobre o desempenho animal, produção e qualidade da carne de novilhos Nelore ou cruzados (½ Aberdeen Angus + ½ Nelore). Os animais foram obtidos pelo acasalamento entre matrizes Nelore com touros Nelore (N) ou Aberdeen Angus (AN). Ambos os grupos genéticos paternos foram selecionados para possuírem diferenças esperadas na progênie (DEP) positivas (+, alto MGA) ou negativas (-, baixo MGA) para acabamento de carcaça (DEPacb). Utilizou-se delineamento em blocos casualizados, em arranjo fatorial 2 (alto x baixo MGA) x 2 (N x AN). Três estudos foram conduzidos. No primeiro foram utilizados 76 animais (14 N+, 20 N-, 20 AN+ e 22 AN-) em sistema de recria a pasto e terminação de novilhos não castrados em confinamento. Neste estudo avaliou-se o desempenho, comportamento alimentar em confinamento, expressão gênica no tecido adiposo subcutâneo (TAS) e características de carcaça. Os animais foram abatidos e imediatamente foram coletadas amostras de tecido adiposo subcutâneo (TAS) para estudo da expressão relativa dos genes ACC, ADIPOQ, FASN, PLIN, PPAR e SREBP1. As carcaças foram avaliadas quali-quantitativamente. Para o segundo estudo, foram utilizados 172 animais (34 N+, 33 N-, 46 AN+ e 59 AN-) em sistemas de recria e terminação a pasto. Neste estudo, foram avaliados ao final da recria o desempenho e características de carcaça por ultrassonografia e ao abate. No terceiro experimento utilizou-se 214 novilhos (48 N+, 53 N-, 51 AN+ e 62 AN-) e avaliou-se características físico-químicas da carne e suas correlações lineares com a espessura de gordura subcutânea (EGS). Foram analisados o pH, cor, perdas por exsudação e cocção (PCo) e força de cisalhamento (FC) em amostras de carne não maturadas e maturada por 14 dias. Foram determinadas as concentrações de umidade e gordura (GO) da carne. O MGA não influenciou (P > 0,05) o desempenho, o comportamento alimentar em confinamento e a expressão relativa dos genes estudados. Animais de alto MGA apresentaram maiores (P < 0,05) EGS, escores de acabamento, de conformação e de distribuição de gordura na carcaça em relação aos animais de baixo MGA. A EGS e expressão relativa dos genes ACC, PLIN, PPAR foram maiores (P < 0,05) no TAS dos animais AN em relação aos N. O MGA não influenciou (P > 0,05) as características físico-químicas da carne. As associações entre EGS e pH, a*, PCo e FC foram significativas (P < 0,05), no entanto, de baixa magnitude. O MGA permite a obtenção de carcaças com maior grau de acabamento sem influenciar o desempenho, comportamento alimentar em confinamento, atributos físico-químicos da carne e expressão relativa dos genes estudados.

Subcutaneous fat deposition in beef cattle is great relevance and the genetic value of breeding breeds can contribute to the understanding and efficient control of this trait in beef production systems. The objective was to evaluate the effects of genetic merit for finishing (GMF) on animal performance, production and meat quality of Nellore or crossbred (½ Aberdeen Angus + ½ Nellore) steers. Animals were obtained by mating Nellore sires with Nellore (N) or Aberdeen Angus (AN) bulls. Both sire genetic groups were selected to have positive (+, high MGA) or negative (-, low GMF) expected progeny differences for backfat thickness (EPDbf). A randomized block design with a factorial arrangement of treatments 2 (high x low GMF) x 2 (N x AN) was employed. Three studies were conducted. In the first, 76 animals (14 N +, 20 N- , 20 AN + and 22 AN-) were used in a pasture growing system and finishing of non-castrated steers in feedlot. In this study, performance, feeding behavior in feedlot, gene expression in subcutaneous adipose tissue (SAT) and carcass characteristics were evaluated. The animals were slaughtered, and SAT samples were collected to study the relative expression of ACC, ADIPOQ, FASN, PLIN, PPAR and SREBP1 genes. Carcasses were evaluated qualitatively and quantitatively. The second study, 172 animals (34 N +, 33 N-, 46 AN + and 59 AN-) were used in growing and finishing pasture-based systems. In this study, performance and carcass characteristics were evaluated at the end of growing phase (by ultrasound) and at slaughter. The third experiment, 214 steers (48 N +, 53 N, 51 AN + and 62 AN-) were used and the physicochemical characteristics of meat and its linear correlations with the subcutaneous fat thickness (SFT) were evaluated. In meat samples prior maturation and matured for 14 days the pH, color (L*, a*, b*) losses by exudation and cooking (CL) and shear force (SF) were analyzed. Meat moisture and intramuscular fat (IMF) concentrations were determined. GMF did not influence (P < 0.05) performance, feedlot behavior and the relative expression of the studied genes. High GMF animals showed greater (P < 0.05) SFT, finishing scores, conformation scores and fat distribution scores in carcass compared to low MGA steers. The SFT and relative expression of ACC, PLIN, PPAR genes were higher (P < 0.05) in SAT of AN steers in relation to N. MGA did not influence (P > 0.05) the physicochemical and meat composition traits. Associations between SFT and attributes of pH, a *, CL and SF were significant (P < 0.05), however, with low magnitude. The GMF makes possible to obtain steers with finishing degree without influencing performance, feeding behavior and physicochemical meat attributes. The difference in fat deposition on carcass obtained by MGA is not related to the relative expression of studied genes.

CONSUMO ALIMENTAR RESIDUAL, NUTRIÇÃO PRÉ E PÓSNATAL E CRESCIMENTO DE BEZERROS NELORE

Objetivou-se com este estudo avaliar o efeito entre o consumo alimentar residual (CAR) de matrizes Nelore e o potencial de crescimento e expressão dos genes de suas progênies, com ambientes nutricionais distintos durante gestações subsequentes, em características de crescimento dos bezerros do nascimento ao desmame. Foram utilizadas 52 matrizes contemporâneas, classificadas como CAR negativo e positivo, submetidas à protocolos de inseminação artificial em tempo fixo aos dois e três anos de idade, com sêmen de mesmo touro da raça Nelore, de CAR negativo. Durante o primeiro experimento as matrizes permaneceram em piquete de Brachiaria brizantha, cultivar Marandu e durante o segundo experimento as matrizes permaneceram em confinamento, com dieta composta por silagem de milho e farelo de soja. As matrizes foram pesadas nos três meses do segundo trimestre gestacional e as progênies foram pesadas ao nascimento e periodicamente cada 28 dias, para o cálculo do peso aos 120 e 210 dias, momento do desmame. Medidas de ultrassonografia de carcaça, nas regiões da garupa, como medida longitudinal para determinação da espessura de gordura subcutânea na garupa (EGG), e medida transversal entre a 12ª e a 13ª costelas para determinação da espessura de gordura subcutânea (EGS) e a área de olho de lombo (AOL), foram tomadas nas matrizes (final do segundo trimestre gestacional) e nos bezerros (momento do desmame), também foi realizada expressão gênica dos bezerros no nascimento e no desmame. O delineamento experimental utilizado foi o inteiramente casualizado e as análises de variância foram realizadas pelo procedimento MIXED do programa SAS. No primeiro experimento, a EGS das matrizes foi significativo (6,32 vs 4,97 mm, P = 0,058 CAR negativo e CAR positivo, respectivamente), para os bezerros as variáveis significativas foram peso ao nascimento (35,8 vs 32,6 kg, P = 0,035 CAR negativo e CAR positivo, respectivamente), peso aos 120 dias (163 vs 149 kg, P = 0,031 CAR negativo e CAR positivo, respectivamente), peso aos 210 dias (262 vs 239 kg, P = 0,033 CAR negativo e CAR positivo, respectivamente), ganho médio diário (1,08 vs 0,99 kg/dia, P = 0,058 CAR negativo e CAR positivo, respectivamente), EGS (2,57 vs 1,91 mm, P = 0,066 CAR negativo e CAR positivo, respectivamente), EGG (6,27 vs 5,04 mm, P = 0,012 CAR negativo e CAR positivo, respectivamente) e produção de bezerros desmamados (272 vs 244 kg/vaca, P = 0,046 CAR negativo e CAR positivo, respectivamente), e a expressão dos genes no nascimento foi significativo PPARG (P = 0,006, bezerros de matrizes CAR positivo), LPL (P = 0,087, bezerros de matrizes CAR positivo) e FABP4 (P = 0,095, bezerros de matrizes CAR positivo), no desmame apenas o gene PPARG foi significativo (P = 0,064, bezerros de matrizes CAR negativo). No segundo experimento as variáveis significativas das matrizes foram peso no 3º mês do segundo trimestre gestacional (639 vs 502 kg, P = 0,089 CAR negativo e CAR positivo, respectivamente) e ganho médio diário (0,84 vs 0,08 mm, P = 0,095 CAR negativo e CAR positivo, respectivamente), a variável significativa nos bezerros foi EGS (2,11 vs 2,83 mm, P = 0,003 CAR negativo e CAR positivo, respectivamente), e a expressão dos genes no nascimento foi significativo SCD1 (P = 0,076, bezerros de matrizes CAR positivo) e ACOX1 (P = 0,094, bezerros de matrizes CAR negativo), no desmame apenas o gene FASN foi significativo (P = 0,097, bezerros de matrizes CAR positivo). A situação nutricional (pré e pós-natal) exerce influência na deposição dos tecidos corporais no início da vida de bovinos da raça Nelore. As matrizes Nelore classificadas como CAR negativo são benéficas para os sistemas de produção brasileiros, pois consomem menos alimentos e produzem bezerros com desempenhos melhores ou semelhantes ao das matrizes classificadas como CAR positivo.

The objective of this study was to evaluate the effect between the residual food consumption (CAR) of Nellore breeds and the growth potential and expression of the genes of their progenies, with different nutritional environments during subsequent pregnancies, on growth characteristics of calves from birth to weaning. Fifty two contemporary breeders, classified as negative and positive RFI, were submitted to fixedtime artificial insemination protocols at two and three years of age, with the same negative Nelore bull semen. During the first experiment the dams remained in Brachiaria brizantha picket, cultivar Marandu and during the second experiment the dams remained in confinement with a diet composed of corn silage and soybean meal. The dams were weighed in the three months of the second gestational trimester and the progenies were weighed at birth and periodically every 28 days to calculate the weight at 120 and 210 days, weaning time. Carcass ultrasound measurements in the croup regions as a longitudinal measure for determination of subcutaneous fat thickness in the croup (FTC) and cross-sectional measurement between the 12th and 13th ribs for determination of subcutaneous fat thickness (SFT) and area loin eye (AOL), were taken in the calves (end of the second trimester of pregnancy) and calves (weaning time), was also performed gene expression of calves at birth and weaning. The experimental design was completely randomized and the analysis of variance was performed by the MIXED procedure of the SAS program. In the first experiment, the SFT of the dams was significant (6.32 vs 4.97 mm, P = 0.058 RFI negative and RFI positive, respectively), for calves the significant variables were birth weight (35.8 vs 32.6 kg, P = 0.035 RFI negative and RFI positive, respectively), weight at 120 days (163 vs 149 kg, P = 0.031 RFI negative and RFI positive, respectively), weight at 210 days (262 vs 239 kg, P = 0.033 RFI negative and positive RFI, respectively), average daily gain (1.08 vs 0.99 kg / day, P = 0.058 negative RFI and positive RFI, respectively), SFT (2.57 vs 1.91 mm, P = 0.066 RFI negative and RFI positive, respectively), FTC (6.27 vs 5.04 mm, P = 0.012 RFI negative and RFI positive, respectively) and production of weaned calves (272 vs 244 kg / cow, P = 0.046 RFI negative and RFI positive, respectively), and gene expression at birth was significant PPARG (P = 0.006, RFI positive calves), LPL (P = 0.087, RFI positive calves) and FABP4 (P = 0.095, RFI positive calf), at weaning only the PPARG gene was significant (P = 0.064, RFI negative calf). In the second experiment the significant variables of the dams were weight in the 3rd month of the second gestational trimester (639 vs 502 kg, P = 0.089 RFI negative and RFI positive, respectively) and average daily gain (0.84 vs 0.08 mm, P = 0.095 RFI negative and RFI positive, respectively), the significant variable in calves was SFT (2.11 vs 2.83 mm, P = 0.003 RFI negative and RFI positive, respectively), and gene expression at birth was significant SCD1 ( P = 0.076, RFI positive calf) and ACOX1 (P = 0.094, RFI negative calf), at weaning only the FASN gene was significant (P = 0.097, RFI positive calf). The nutritional situation (pre and postnatal) influences the deposition of body tissues in the early life of Nellore cattle. Nellore breeders classified as negative CAR are beneficial for Brazilian production systems, as they consume less food and produce calves with better or similar performances to the breeders classified as positive CAR.

Adipose-derived stem cells (ADSC) in the viability of a random pattern dorsal skin flap in rats

To evaluate the viability of random pattern dorsal skin flaps in rats after injection of adipose-derived stem cells (ADSC). Thirty five adult male Wistar EPM rats (weight 250-300 g) were distributed, at random, in two groups. I- Control (flap elevation with injection of saline solution) with fifteen animals and II- Experimental (flap elevation with injection of ADSC ) with fifteen animal. The ADSC were isolated from others five adult male rats. A dorsal skin flap measuring 10x4 cm was raised and a plastic barrier was placed between the flap and its bed in both groups and the injection (cells or saline solution) were perfomed immediately after the surgery. The percentage of flap necrosis was measured on the seventh postoperative day. The ADSC were able to replicate in our culture conditions. We also induced their adipogenic, osteogenic and chondrogenic differentiation to verify their mesenchymal stem cells potentiality in vitro. The results were statistically significant showing that the ADSC decreased the area of necrosis (p<0.05). CONCLUSIONS: The cells demonstrated adipogenic, osteogenic and chondrogenic differentiation potential in vitro. The administration of adipose-derived stem cells was effective to increase the viability of the random random pattern dorsal skin flaps in rats.(AU)