Name: Autor em destaque: Melhorando a nutrição e a produção de bovinos de corte com foco na eficiência alimentar e nas características de qualidade da carne por meio de ensaios bioquímicos e moleculares
Uploaded: 2024-07-12T13:50:00
Description: This study investigated muscle tissue in Bos indicus and crossbred bulls to explain differences in meat quality traits. Carcass traits, meat quality ...

Esta pesquisa foi financiada pela FAPESP (processos 2023/05002-3; 2023/02662-2 e 2024/09871-9), CAPES (código de financiamento 001), CNPq (304158/2022-4) e pelo PROPE (IEPe-RC processo número 149) da Faculdade de Medicina Veterinária e Zootecnia da Universidade Estadual Paulista.

Todos os procedimentos com animais obedeceram às normas éticas de pesquisa estabelecidas pelo Comitê de Ética no Uso de Animais (CEUA) da Universidade Estadual Paulista Júlio de Mesquita Filho - UNESP Campus de Botucatu, sob o protocolo 0171/2018.
1. Animais experimentais
<ol><li>Terminar 30 touros Nelore (Bos indicus) e 30 touros F1 Angus-Nelore (Bos taurus × Bos indicus), com idade entre 20 e 24 meses, em confinamento. Alojar os dois grupos...

Avaliação de características de qualidade da carne em touros Bos indicus e mestiços 

Durante a avaliação da carcaça, é crucial medir com precisão as características de crescimento e qualidade após um período de resfriamento de 48 h para obter dados consistentes e comparáveis. Os dois modelos biológicos exibiram características de carcaça divergentes, particularmente HCW, REA e BFT, que são consistentes com os achados relatados em outros estudos. A média de PCS de touros Nelore está alinhada com as preferências do mercado brasileiro, que prioriza maior produção de carne por unidade anima...

O Brasil possui o maior rebanho bovino comercial do mundo, com aproximadamente 220 milhões de animais e sendo o segundo maior produtor de carne, produzindo mais de 9 milhões de toneladas métricas de carcaça equivalente anualmente1. O setor de produção de bovinos de corte contribui significativamente para o sistema agrícola nacional, com faturamento anual total superior a R$ 55 bilhões. Desde 2004, o Brasil tem sido um ator chave no comércio global de carne, exportando para mais de 180 países, o que representa ~ 50% do comércio mundial de carne2.
A maciez da carne destaca-se como o atributo primordial da qualidade que influencia a satisfação do consumidor e o consumo de carne3. Ao empregar métodos bioquímicos e objetivos para medir a maciez da carne, os pesquisadores pretendem fornecer informações valiosas sobre fatores como genética animal, técnicas de processamento e condições de armazenamento, melhorando a qualidade e a consistência dos produtos cárneos para os consumidores. Essas informações são úteis porque a maciez da carne ganhou cada vez mais importância na tomada de decisão do consumidor durante as compras. Além disso, a avaliação da maciez da carne fornece informações valiosas para o controle de qualidade nas indústrias de produção e processamento de carne. Ao monitorar consistentemente a maciez, os produtores podem garantir que os produtos à base de carne atendam aos padrões e especificações desejados. Nesse contexto, os produtores brasileiros de gado de corte estão adotando progressivamente sistemas intensivos de confinamento com animais mestiços para aumentar o giro de capital. Esse sistema é responsável por cerca de 10% das toneladas de carcaça produzidas anualmente no Brasil 4,5.
A crescente demanda por melhor qualidade da carne pelos consumidores levou os produtores de gado de corte a cruzar com raças europeias, principalmente Aberdeen Angus6. Essa estratégia visa produzir híbridos F1 Angus-Nelore, conhecidos por desempenho superior, características de carcaça desejáveis e melhor qualidade da carne em comparação com animais zebuínos puros 7,8. Nas regiões tropicais do Brasil, é prática comum a utilização de animais não castrados (touros) de maturidade avançada em fazendas de terminação, potencialmente comprometendo atributos de qualidade da carne, como cor, marmoreio e maciez. Destaca-se que 95% dos animais terminados em confinamento brasileiros são machos, sendo 73% Nelore, seguidos por 22% de animais mestiços e 5% de outros genótipos 9,10.
Compreender os mecanismos bioquímicos subjacentes à maciez da carne é crucial para melhorar a qualidade da carne. Um aspecto fundamental é a proteólise post-mortem, que afeta a integridade estrutural das fibras musculares11. O índice de fragmentação miofibrilar (MFI) é um ensaio bioquímico amplamente utilizado que quantifica a extensão da degradação miofibrilar, fornecendo informações sobre a maciez da carne12. O método MFI envolve a medição da fragmentação das proteínas miofibrilares, que se correlaciona diretamente com a maciez da carne. Este ensaio complementa as avaliações tradicionais da qualidade da carne e oferece uma compreensão mais profunda dos processos bioquímicos que contribuem para as variações na maciez da carne.
Nesse contexto, o presente estudo investigou o músculo esquelético de touros Bos indicus versus touros mestiços (Bos taurus × Bos indicus) terminados em confinamento, com o objetivo de explicar diferenças nas características de qualidade da carne.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Acetone</td>
			<td>Merk, Darmstadt, Germany</td>
			<td>CAS 67-64-1 | 100014</td>
			<td>solutions used for the electrophoretic separations</td>
		</tr>
		<tr>
			<td>Anti-MYH-1 Antibody</td>
			<td>Merk, Darmstadt, Germany</td>
			<td>MABT846</td>
			<td>Rat soleus</td>
		</tr>
		<tr>
			<td>Anti-Myosin antibody</td>
			<td>Abcam, Massachusetts, United States</td>
			<td>ab37484</td>
			<td>Myosin heavy chain</td>
		</tr>
		<tr>
			<td>Anti-Myosin-2 (MYH2) Antibody</td>
			<td>Merk, Darmstadt, Germany</td>
			<td>MABT840</td>
			<td>Extensor digitorum longus (EDL)</td>
		</tr>
		<tr>
			<td>Biological oxygen demand (BOD) incubator</td>
			<td>TECNAL, S&#227;o Paulo, Brazil</td>
			<td>TE-371/240L</td>
			<td>Meat aging</td>
		</tr>
		<tr>
			<td>Chloroform; absolute analytical reagent</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>67-66-3</td>
			<td>Intramuscular fat</td>
		</tr>
		<tr>
			<td>CIELab system</td>
			<td>Konica Minolta Sensing, Tokyo, Japan</td>
			<td>CR-400 colorimeter</td>
			<td>Meat color</td>
		</tr>
		<tr>
			<td>Coomassie Blue</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>C.I. 42655)</td>
			<td>Myosin heavy chain</td>
		</tr>
		<tr>
			<td>Electric oven</td>
			<td>Ven&#226;ncio Aires, Rio Grande do Sul, Brazil</td>
			<td></td>
			<td>Meat tenderness</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Merk, Darmstadt, Germany</td>
			<td>64-17-5</td>
			<td>solutions used for the electrophoretic separations</td>
		</tr>
		<tr>
			<td>Ethylenediaminetetraacetic acid</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>60-00-4</td>
			<td>Post-mortem proteolysis</td>
		</tr>
		<tr>
			<td>Glass flasks</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td></td>
			<td>solutions used for the electrophoretic separations</td>
		</tr>
		<tr>
			<td>Glycine</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>G6761</td>
			<td>Myosin heavy chain</td>
		</tr>
		<tr>
			<td>Infrared spectroscopy - FoodScan</td>
			<td>Foss NIRSystems, Madson, United States</td>
			<td>FoodScan&#8482; 2</td>
			<td>Intramuscular fat</td>
		</tr>
		<tr>
			<td>Magnesium chloride</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>&#160;7786-30-3</td>
			<td>Post-mortem proteolysis</td>
		</tr>
		<tr>
			<td>Mercaptoetanol</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>M6250</td>
			<td>Myosin heavy chain</td>
		</tr>
		<tr>
			<td>Methanol, absolute analytical reagent</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>67-56-1</td>
			<td>Intramuscular fat</td>
		</tr>
		<tr>
			<td>pH meter</td>
			<td>LineLab, S&#227;o Paulo, Brazil</td>
			<td>AKLA 71980</td>
			<td>Meat pH</td>
		</tr>
		<tr>
			<td>PlusOne 2-D Quant Kit</td>
			<td>GE Healthcare Product</td>
			<td>Code 80-6483-56</td>
			<td>Post-mortem proteolysis</td>
		</tr>
		<tr>
			<td>Polypropylene</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td></td>
			<td>solutions used for the electrophoretic separations</td>
		</tr>
		<tr>
			<td>Potassium chloride</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>7447-40-7</td>
			<td>Post-mortem proteolysis</td>
		</tr>
		<tr>
			<td>Potassium phosphate</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>P0662</td>
			<td>Post-mortem proteolysis</td>
		</tr>
		<tr>
			<td>R software</td>
			<td>Vienna, Austria</td>
			<td>version 3.6.2</td>
			<td>Data analysis</td>
		</tr>
		<tr>
			<td>Sodium azide</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>26628-22-8</td>
			<td>Post-mortem proteolysis</td>
		</tr>
		<tr>
			<td>Sodium dodecyl sulfate&#160;(SDS)</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>822050</td>
			<td>Myosin heavy chain</td>
		</tr>
		<tr>
			<td>Spectrophotometer</td>
			<td>Perkin Elmer, Shelton, United States</td>
			<td>Perkin Elmer 
			Lambda 25 UV/Vis</td>
			<td>Post-mortem proteolysis</td>
		</tr>
		<tr>
			<td>Statistical Analysis System</td>
			<td>SAS, Cary, North Carolina, United States</td>
			<td>version 9.1,</td>
			<td>Data analysis</td>
		</tr>
		<tr>
			<td>Texture Analyzer</td>
			<td>AMETEK Brookfield, Massachusetts, United 
			States</td>
			<td>CTX</td>
			<td>Meat tenderness</td>
		</tr>
		<tr>
			<td>Tris(hydroxymethyl)aminomethane</td>
			<td>Sigma-Aldrich, Missouri, United States</td>
			<td>77-86-1</td>
			<td>Myosin heavy chain</td>
		</tr>
		<tr>
			<td>Ultrafreezer</td>
			<td>Indrel Scientific, Londrina, Paran&#225;, Brazil.</td>
			<td>INDREL IULT 335 D - LCD</td>
			<td>Sample storage</td>
		</tr>
		<tr>
			<td>Ultrapure water</td>
			<td colspan="2">Elga PURELAB Ultra Ionic system</td>
			<td>solutions used for the electrophoretic separations</td>
		</tr>
		<tr>
			<td>Ultra-Turrax high shear mixer</td>
			<td>Marconi &#8211; MA102/E, Piracicaba, S&#227;o Paulo, Brazil</td>
			<td></td>
			<td>Post-mortem proteolysis</td>
		</tr>
	</tbody>
</table>

exploring the longissimus muscle: unraveling its correlation with meat quality in bos indicus and crossbred bulls

Este estudo investigou o tecido muscular em touros Bos indicus e mestiços para explicar as diferenças nas características de qualidade da carne. Características de carcaça, parâmetros de qualidade da carne e investigações bioquímicas e moleculares de proteínas miofibrilares são descritas. Métodos para avaliar pH, gordura intramuscular (IMF), cor da carne (L*, a*, b*), perdas de água, maciez e ensaios de biologia molecular foram delineados. São descritos procedimentos específicos detalhando a calibração, a preparação da amostra e a análise de dados para cada método. Isso inclui técnicas como espectroscopia de infravermelho para conteúdo de IMF, avaliação objetiva de maciez e separação eletroforética de isoformas de MyHC.
Os parâmetros de cor foram destacados como ferramentas potenciais para prever a maciez da carne bovina, uma característica de qualidade crucial que influencia as decisões do consumidor. O estudo empregou o método da força de cisalhamento de Warner-Bratzler (WBSF), revelando valores de 4,68 e 4,23 kg para Nelore e Angus-Nelore (P < 0,01), respectivamente. As perdas totais por cozimento e as análises bioquímicas, incluindo o índice de fragmentação miofibrilar (MFI), forneceram informações sobre as variações de maciez. Tipos de fibras musculares, particularmente isoformas de cadeia pesada de miosina (MyHC), foram investigados, com notável ausência da isoforma MyHC-IIb nos animais zebuínos estudados. A relação entre MyHC-I e maciez da carne revelou achados divergentes na literatura, destacando a complexidade dessa associação. No geral, o estudo fornece informações abrangentes sobre os fatores que influenciam a qualidade da carne em touros Bos indicus e mestiços (Bos taurus × Bos indicus), oferecendo informações valiosas para a indústria de carne bovina.

Brazil has the largest commercial cattle herd globally, numbering approximately 220 million animals and ranking as the second-largest meat producer, yielding over 9 million metric tons of carcass equivalent annually1. The beef cattle production sector significantly contributes to the national agricultural system, with total annual sales surpassing R$ 55 billion. Since 2004, Brazil has been a key player in the global meat trade, exporting to over 180 countries, which represents ~50% of the world meat trade2.
Meat tenderness stands out as the paramount quality attribute influencing consumer satisfaction and meat consumption3.&#160;By employing biochemical and objective methods to measure meat tenderness, researchers aim to provide valuable insights into factors such as animal genetics, processing techniques, and storage conditions, ultimately enhancing the quality and consistency of meat products for consumers. Such information is useful because meat tenderness has gained increased importance in consumer decision-making during purchases. Moreover, meat tenderness assessment provides valuable information for quality control in meat production and processing industries. By consistently monitoring tenderness, producers can ensure that meat products meet desired standards and specifications. In this context, Brazilian beef cattle producers are progressively embracing intensive feedlot systems with crossbred animals to enhance capital turnover. This system accounts for about 10% tons of carcass produced annually in Brazil4,5.
The escalating demand for improved meat quality by consumers has prompted beef cattle producers to crossbreed with European breeds, primarily Aberdeen Angus6. This strategy aims to produce F1 Angus-Nellore hybrids, known for superior performance, desirable carcass traits, and enhanced meat quality compared to pure zebu animals7,8. In the tropical regions of Brazil, it is common practice to utilize non-castrated animals (bulls) of advanced maturity in finishing farms, potentially compromising meat quality attributes such as color, marbling, and tenderness. Notably, a survey reveals that 95% of animals finished in Brazilian feedlots are males, with 73% being Nellore, followed by 22% crossbred animals and 5% other genotypes9,10.
Understanding the biochemical mechanisms underlying meat tenderness is crucial for improving meat quality. One key aspect is postmortem proteolysis, which affects the structural integrity of muscle fibers11. The myofibril fragmentation index (MFI) is a widely used biochemical assay that quantifies the extent of myofibril degradation, providing insights into the tenderness of meat12. The MFI method involves measuring the fragmentation of myofibrillar proteins, which directly correlates with meat tenderness. This assay complements traditional meat quality assessments and offers a deeper understanding of the biochemical processes that contribute to variations in meat tenderness.
In this context, the present study investigated the skeletal muscle of Bos indicus versus crossbred (Bos taurus &#215; Bos indicus) bulls finished in feedlot, aiming to explain differences in meat quality traits.

Autor em destaque: Melhorando a nutrição e a produção de bovinos de corte com foco na eficiência alimentar e nas características de qualidade da carne por meio de ensaios bioquímicos e moleculares 

Explorando o músculo longuíssimo: desvendando sua correlação com a qualidade da carne em touros Bos indicus e mestiços

Our research involves improving beef cattle nutrition and production, with a significant focus on feed efficiency and meat quality traits. Recently, we have investigated several feed efficiency index of cattle. For example, the residual feed intake, to understand their impact on carcass and meat quality traits.We applied biochemical and molecular assays, including proteomics, lipidomics, and metabolomic approach, to investigate beef cattle nutrition strategy, particularly in feedlot diets. Besides, we also aim to explain the difference in meat quality traits between cattle breeds and sex class. The last years, our findings explored the muscle tissue proteome of beef cattle, which allow us to a better understand of our nutritional strategy used in the livestock production systems.Our lab will focus on omic related analytic technologies and bioinformatics to identify proteins related to growth and meat quality traits of livestock species, since they are the main constituents of muscle tissue and also they're responsible for regulation of the main metabolic pathways.

A Tabela 1 apresenta as características de carcaça dos dois grupos genéticos investigados neste estudo. Notavelmente, foram identificadas diferenças (P < 0,01) em PCS, REA e BFT, com animais mestiços exibindo maiores valores, sugerindo um efeito de heterose.
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td>Variável¹</td>
			<td>Nelore</td>
			<td>F1 Angus x Nelore</td>
			<td>SEM</td>
			<td>Valor de p</td>
		</tr><tr><t...

Investigamos o tecido muscular esquelético em Bos indicus e touros mestiços para explicar as diferenças nas características de qualidade da carne. A força de cisalhamento Warner-Bratzler (WBSF) variou de 4,7 kg a 4,2 kg. As isoformas de cadeia pesada da miosina revelaram diferenças entre os animais, e o índice de fragmentação da miofibrila forneceu mais informações sobre as variações da maciez (WBSF).

This study investigated muscle tissue in Bos indicus and crossbred bulls to explain differences in meat quality traits. Carcass traits, meat quality ...

exploring-longissimus-muscle-unraveling-its-correlation-with-meat

Research

JoVE Journal

Biology

Exploring the Longissimus Muscle: Unraveling its Correlation with Meat Quality in Bos indicus and Crossbred Bulls

This study investigated muscle tissue in Bos indicus and crossbred bulls to explain differences in meat quality traits. Carcass traits, meat quality parameters, and biochemical and molecular investigations of myofibrillar proteins are described. Methods for evaluating pH, intramuscular fat (IMF), meat color (L*, a*, b*), water losses, tenderness, and molecular biology assays have been outlined. Specific procedures detailing calibration, sample preparation, and data analysis for each method are described. These include techniques such as infrared spectroscopy for IMF content, objective tenderness assessment, and electrophoretic separation of MyHC isoforms.
Color parameters were highlighted as potential tools for predicting beef tenderness, a crucial quality trait influencing consumer decisions. The study employed the Warner-Bratzler shear force (WBSF) method, revealing values of 4.68 and 4.23 kg for Nellore and Angus-Nellore (P &#60; 0.01), respectively. Total cooking losses and biochemical analyses, including myofibril fragmentation index (MFI), provided insights into tenderness variations. Muscle fiber types, particularly myosin heavy chain (MyHC) isoforms, were investigated, with a notable absence of MyHC-IIb isoform in the studied Zebu animals. The relationship between MyHC-I and meat tenderness revealed divergent findings in the literature, highlighting the complexity of this association. Overall, the study provides comprehensive insights into the factors influencing meat quality in Bos indicus and crossbred (Bos taurus &#215; Bos indicus) bulls, offering valuable information for the beef industry.

We investigated skeletal muscle tissue in Bos indicus and crossbred bulls to explain differences in meat quality traits. Warner-Bratzler shear force (WBSF) was found to range from 4.7 kg to 4.2 kg. Myosin heavy chain isoforms revealed differences between animals, and myofibril fragmentation index provided further insights into tenderness (WBSF) variations.

Biologia

Comprehensive Evaluation of Carcass Traits and Meat Quality Parameters in Beef Cattle

To begin place the longissimus thoracis, or LT muscle, on the working platform. Measure the rib eye area in the 12th and 13th rib interface using a small grid with a dot in the middle. Sum all squares within the rib eye, tracing perimeter, and those along the contour, passing through the middle dot.To measure the back fat thickness, place the caliper at a right angle to the line of the subcutaneous fat from the interface point between the 12th and 13th ribs. Calibrate the pH meter probe with pH-4 and 7 buffers at room temperature. Measure the meat pH at three locations of the longissimus thoracis, or LT muscle.Manually record the data readings, export the data sheet, and calculate the average of the three readings. Homogenize the LT muscle in a methanol chloroform solution for two minutes. Centrifuge the muscle suspension at 700 G for 10 minutes at 20 degrees Celsius to segregate the hydrophilic, solid, and hydrophobic phases.Aspirate the alcoholic layer. Filter the homogenate through filter paper on a funnel with a slight suction. Apply pressure with the bottom of a beaker when the residue becomes dry.Transfer the filtrate to a 500 milliliter graduated cylinder and let it stand for a few minutes. Record the volume of the chloroform layer. Place the sample in an oven at 110 degrees Celsius until complete solvent evaporation.Then, cool the sample in a desiccator overnight. The next day, reweigh the beaker, and determine the intramuscular fat content by calculating the difference between the initial and final weights of the beaker. To calibrate the colorimeter, place the white calibration plate near the middle of the plate.Confirm the calibration once the lamp flashes three times. After 30 minutes of blooming at four degrees Celsius, obtain color readings from three different locations on the LT muscle. Compute the average from three measurements.To determine the purge loss of the beef loin section, measure the variance between the initial weight before freezing, the weight after freezing and thawing, and the final weight. To gauge the water-holding capacity, first, weigh the meat sample before subjecting it to a pressure of 10 kilograms for five minutes. Then, weigh the meat sample again and assess the water holding capacity by calculating the weight differences.Position LT muscles on a grid attached to a glass refractory, place the digital thermometer in the sample, and cook the muscles in an industrial electric oven until reaching a final temperature of 71 degrees Celsius. After cooling, weigh the sample, and refrigerate at four degrees Celsius for 24 hours. Use the given formula to calculate the cooking losses.To determine drip loss, weigh the refractory before cooking the sample. Place the samples on a grid over a glass refractory to allow drainage of meat juices and fat during cooking. After cooking, again, weigh the refractory.Weigh only the sample before and after cooking to determine evaporation loss. After recording raw and cooked weights, calculate the percentage of drip loss as the weight of drip after cooking, divided by the weight of the thawed meat sample. Calculate the evaporation loss percentage using the given formula.For the determination of Warner-Bratzler shear force, section eight cores using a texture analyzer equipped with a Warner-Bratzler blade and V-shaped cutting edge. After excluding the low and high extremes, report the results as the average of six values per sample. Homogenize three grams of LT muscles for 30 seconds in a potassium buffer at two degrees Celsius.Then, centrifuge the sample at 1000 G for 15 minutes at four degrees Celsius, and resuspend the pellet in 10 volumes of isolating medium using a stir rod. Again, centrifuge and resuspend the pellet in 2.5 volumes of isolating medium. Pass the suspension through a polyethylene strainer to separate the connective tissue and debris.Use an additional 2.5 volumes of isolating medium to allow the myofibrils to pass through the strainer. Next, dilute an aliquot of myofibril suspension with an isolating medium. Determine the protein concentration of the suspension of myofibrils using the biuret method.Immediately, measure the absorbance at 540 nanometers. Multiply the absorbance value by 200 to obtain the myofibrillar fragmentation index for each value. Significant differences in hot carcass weight, rib eye area, and back fat thickness were observed with crossbred animals showing higher values, indicating a heterosis effect.Meat quality traits comparisons showed no significant differences in meat pH, purge loss, extractable volume percentage, and cooking loss between Nellore and F1 Angus Nellore bulls. However, crossbred bulls had higher intramuscular fat, yellowness, water holding capacity, and myofibrillar fragmentation index. The meat of Nellore bulls showed reduced tenderness with higher Warner-Bratzular shear force and drip loss, indicating it is tougher than the meat of F1 Angus Nellore bulls.