Metalloproteomic analysis of liver proteins isolated from broilers fed with different sources and levels of copper and manganese.

Supplementing minerals beyond dietary requirements can increase the risk of toxicity and mineral excretion, making the selection of more bioavailable sources crucial. Thus, this work aimed to use metalloproteomics tools to investigate possible alterations in the hepatic proteome of broilers fed with diets containing two sources (sulfate and hydroxychloride) and two levels of copper (15 and 150 ppm) and manganese (80 and 120 ppm), totaling four treatments: low Cu/Mn SO4, high Cu/Mn SO4, low Cu/Mn (OH)Cl and high Cu/Mn (OH)Cl. The difference in abundance of protein spots and copper and manganese concentrations in liver and protein pellets were analyzed by analysis of variance with significance level of 5%. The Cu and Mn concentrations determined in liver and protein pellets suggested greater bioavailability of hydroxychloride sources. We identified 19 Cu-associated proteins spots, 10 Mn-associated protein spots, and 5 Cu and/or Mn-associated protein spots simultaneously. The analysis also indicated the induction of heat shock proteins and detoxification proteins in broilers fed with high levels of copper and manganese, suggesting the involvement of these proteins in metal tolerance and stress.

Proteomic Study of Broiler Plasma Supplemented with Different Levels of Copper and Manganese from Different Sources.

The aim of the present study was to evaluate the differential expression of plasma proteins in broiler chickens supplemented with different sources (sulfates and hydroxychlorides) and levels of copper (15 and 150 mg kg-1) and manganese (80 and 120 mg kg-1). For this, plasma samples from 40 broiler chickens were used, divided into four experimental groups: S15-80 (15 ppm CuSO4 and 80 ppm MnSO4), S150-120 (150 ppm CuSO4 and 120 ppm MnSO4), H15-80 (15 ppm Cu(OH)Cl and 80 ppm Mn(OH)Cl), and H150-120 (150 ppm Cu(OH)Cl and 120 ppm Mn(OH)Cl). From plasma samples obtained from each bird from the same treatment, four pools were made considering 10 birds per group. Plasma proteome fractionation was performed by 2D-PAGE. Concentrations of the studied minerals were also evaluated in both plasma and protein pellet samples. A higher concentration of Cu and Mn was observed in the plasma and protein pellets of groups that received higher mineral supplementation levels compared to those receiving lower levels. Mn concentrations were higher in plasma and protein pellets of the hydroxychloride-supplemented groups than the sulfate-supplemented groups. Analysis of the gels revealed a total of 40 differentially expressed spots among the four treatments. Supplementation with different sources of minerals, particularly at higher levels, resulted in changes in protein regulation, suggesting a potential imbalance in homeostasis.

Feedlot diets containing different starch levels and additives change the cecal proteome involved in cattle's energy metabolism and inflammatory response.

Diets for feedlot cattle must be a higher energy density, entailing high fermentable carbohydrate content. Feed additives are needed to reduce possible metabolic disorders. This study aimed to analyze the post-rumen effects of different levels of starch (25%, 35%, and 45%) and additives (monensin or a blend of essential oils and exogenous α-amylase) in diets for Nellore feedlot cattle. The cecum tissue proteome was analyzed via two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and then differentially expressed protein spots were identified with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The use of blends of essential oils associated with α-amylase as a feed additive promoted the upregulation of enzymes such as triosephosphate isomerase, phosphoglycerate mutase, alpha-enolase, beta-enolase, fructose-bisphosphate aldolase, pyruvate kinase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), L-lactate dehydrogenase B, L-lactate dehydrogenase A chain, L-lactate dehydrogenase, and ATP synthase subunit beta, which promote the degradation of carbohydrates in the glycolysis and gluconeogenesis pathways and oxidative phosphorylation, support pyruvate metabolism through the synthesis of lactate from pyruvate, and participate in the electron transport chain, producing ATP from ADP in the presence of a proton gradient across the membrane. The absence of proteins related to inflammation processes (leukocyte elastase inhibitors) in the cecum tissues of animals fed essential oils and amylase may be because feed enzymes can remain active in the intestine and aid in the digestion of nutrients that escape rumen fermentation; conversely, the effect of monensin is more evident in the rumen and less than 10% results in post-ruminal action, corroborating the hypothesis that ionophore antibiotics have a limited effect on the microbiota and intestinal fermentation of ruminants. However, the increase in starch in these diets promoted a downregulation of enzymes linked to carbohydrate degradation, probably caused by damage to the cecum epithelium due to increased responses linked to inflammatory injuries.

Effect of energy supplementation on intake, digestibility of diets and performance of grazing lambs during the rainy season.

This study aimed to evaluate different energy sources in multiple supplements on performance, intake, and digestibility of Santa Ines sheep grazing urochloa grass (Urochloa mosambicensis) during the rainy season. The experimental area was divided into seven paddocks of 4 ha each, with an average of dry matter (DM) availability of 3.21 tn/ha. A completely randomized design was carried out, in which there were four treatments, and each treatment was repeated six times. Twenty-four intact lambs (average: 32.0 kg of body weight) were supplemented with a mineral mixture, the control group (MM), mesquite pod meal (MPM), wheat bran (WB), or sorghum grain (SG) as energy sources. The digestibility of DM and crude protein (CP) in MPM and WB is higher than that in MM and SG groups. Neutral detergent fiber (NDF) digestibility was similar between supplemented lambs, and it was higher than the MM. The supplementation promoted higher weight gain than in the control group (0.126 vs. 0.061 g/day, respectively; P < 0.001). The supplementation increased the DM, and CP intake. The NDF intake only increased in the WB group. The CP digestibility was higher for the MPM and WB groups than that for MM and SG ones (P < 0.001). Sheep supplementation in the rainy season increased the average daily gain (ADG). Any supplement tested in the present study can be used during the rainy season. The choice for the supplement will depend on the availability and costs of the mesquite pod meal, sorghum grain, or wheat bran.

Meat Quality and Muscle Tissue Proteome of Crossbred Bulls Finished under Feedlot Using Wet Distiller Grains By-Product.

Wet distiller grains (WDG) are a corn by-product rich in protein and fiber that can be used in feedlot diets. This study evaluated F1 Angus-Nellore bulls fed on a control diet vs. WDG (n = 25/treatment). After a period of 129 days on these feeds, the animals were slaughtered and Longissimusthoracis samples were collected for both a meat quality evaluation and gel-based proteomic analyses. A greater ribeye area (99.47 cm²) and higher carcass weight (333.6 kg) (p < 0.05) were observed in the WDG-finished cattle compared to the control (80.7 cm²; 306.3 kg). Furthermore, there were differences (p < 0.05) in the intramuscular fat between the WDG and control animals (IMF = 2.77 vs. 4.19%), which led to a significant decrease (p < 0.05) in saturated fatty acids (FA). However, no differences (p > 0.10) were observed in terms of tenderness, evaluated using Warner-Bratzler shear force (WBSF). The proteomic and bioinformatic analyses revealed substantial changes in the biological processes, molecular functions, and cellular components of the WDG-finished cattle compared to the control. Proteins related to a myriad of interconnected pathways, such as contractile and structural pathways, energy metabolism, oxidative stress and cell redox homeostasis, and transport and signaling. In this experiment, the use of WDG supplementation influenced the protein expression of several proteins, some of which are known biomarkers of beef quality (tenderness and color), as well as the protein-protein interactions that can act as the origins of increases in muscle growth and reductions in IMF deposition. However, despite the effects on the proteome, the tenderness, evaluated by WBSF, and fatty acid profile were not compromised by WDG supplementation.

Comparison of Dental Carcass Maturity in Non-Castrated Male F1 Angus-Nellore Cattle Finished in Feedlot.

Dental classification of carcasses is used as a parameter of cattle maturity at slaughter, and it can influence carcass and meat quality traits. Brazilian beef-packing companies use the number of permanent incisor (PI) teeth as a parameter for bonus and certification of carcasses with superior quality. However, when non-castrated male such as F1 Angus-Nellore (Bos taurus×Bos indicus) are slaughtered, only animals without PI teeth are subsidized by the breed association. We evaluated these animals finished in feedlot for 180 days with zero versus two PI teeth on the carcass and meat quality traits. At the time of slaughter, 88 carcasses were selected, forming two treatments according to dental carcass maturity (0 versus 2 PI teeth; 44 animals per category). It was demonstrated that the number of PI teeth (0 versus 2 PI) did not influence (p>0.05) carcass (weights, yield, cooling loss, ribeye area and the backfat thickness) and meat quality traits (Longissimus thoracis chemical composition, color, cooking losses, shear force and pH). Thus, dental carcass maturity (zero versus two PI teeth) does not influence non-castrated male F1 Angus-Nellore finished in feedlot for 180 days. This is the first study to demonstrate that carcasses of non-castrated male F1 Angus-Nellore with two PI teeth should be subsidized in a similar way to those with zero PI teeth. Moreover, Brazilian beef-packing companies could produce heavier and leaner carcasses of acceptable quality though the use of crossbred cattle such as non-castrated F1 Angus Nellore.

Intake, digestibility, milk yield and composition, and ingestive behavior of cows supplemented with byproducts from biodiesel industry.

This trial aimed to analyze the effects of including three byproducts from the biodiesel industry on the intake, digestibility, milk yield and composition, and feeding behavior of lactating cows. Eight crossbred Holstein-Zebu lactating cows with average body weight 525 ± 18.5 kg and average milk yield of 8 ± 1.45 kg day-1 were assigned to four treatments (diets) in a double-Latin square design, as follows: a diet based on corn- and soybean meal-based concentrate and three diets with 20% inclusion of byproducts from the biodiesel industry (cottonseed cake, sunflower meal, and castor bean cake) on a total dry matter basis. The cows were housed in individual covered stalls with concrete floor equipped with individual concrete troughs for feeding and automatic drinkers, and fed diets containing 60% sugarcane and 40% concentrate. The inclusion of the byproducts in the diet changed the intake, digestibility of some nutritional components, milk yield and composition, and feeding behavior of lactating cows. The use of cottonseed cake and sunflower meal in the diet increased milk yield, and fat-corrected milk yield; while the use of castor bean cake reduced the intake, digestibility of dry matter and total digestible nutrients, milk yield, and fat-corrected milk yield. The inclusion of byproducts from the biodiesel industry in the diets did not change the fat, lactose, total solids, and solids-not-fat of milk. Therefore, the cottonseed cake and sunflower meal can be included at up to 20% of the total diet.

Metalloproteomic approach of mercury-binding proteins in liver and kidney tissues of Plagioscion squamosissimus (corvina) and Colossoma macropomum (tambaqui) from Amazon region: Possible identification of mercury contamination biomarkers.

Fish is an important source of protein, vitamins, and minerals. However, this food is also a major source of human exposure to toxic contaminants such as mercury. Thus, this paper aimed to evaluate mercury-binding proteins for possible application as biomarkers of mercury contamination in hepatic and renal tissues of Plagioscion squamosissimus (carnivorous fish) and Colossoma macropomum (omnivorous fish) from the Amazon region using metalloproteomic approach. The proteome of hepatic and renal tissues of fish species was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and the mercury concentrations in protein spots were determined by graphite furnace atomic absorption spectrometry (GFAAS). Finally, the protein spots associated to mercury were characterized by electrospray ionization mass spectrometry (ESI-MS/MS). The activity of antioxidant enzymes (SOD, CAT, GPx, and GST) and lipid peroxidation (LPO) were also determined. The results showed that the highest concentrations of mercury were found in the carnivorous species (P. squamosissimus) and that the accumulation pattern of this metal was higher in hepatic tissues than in renal tissues for both species. A tendency was observed for greater enzymatic activity in the hepatic and renal tissues of P. squamosissimus, the species with the highest concentration of mercury. Only GPx activity in the kidney and GST in the liver were lower for the P. squamosissimus species, and this finding can be explained by the interaction of mercury with these enzymes. The data obtained by ESI-MS/MS allowed for the characterization of the protein spots associated to mercury, revealing proteins involved in energy metabolism, biomolecules transport, protein synthesis and degradation, cell differentiation, gene regulation, and the antioxidant system. The results obtained in the present study can contribute to understanding the physiological processes underlying mercury toxicity and have provided new perspectives on possible candidates for mercury contamination biomarkers in fish.

Inclusion of licuri meal in the diet of pasture dairy cows.

The objective of this study was to evaluate the inclusion of increasing levels of licuri meal as a substitute for soybean bran in the diet of pasture-based dairy cows and its effects on the intake, nutrient digestibility, and milk production and composition. Eight crossbred Holstein/Zebu cows were used in two 4 × 4 Latin square plots. The diets contained increasing percentage of licuri meal (0.0, 4.16, 7.09, and 9.45%) in the total diet, replacing soybean bran. The inclusion of licuri meal in the diet affected total dry matter intake and dry matter intake in relation to body weight, which presented a quadratic effect. The same behavior was observed for the consumption of EE, CP, TDN, and NDFap. The NFC consumption showed a linear decrease with the addition of the by-product in the diet. There was no influence of the inclusion of the licuri meal on the digestibility of the DM, CP, NFC, and TDN, while the digestibility of the NDFap and EE presented increasing linear effect. There was a reduction of milk production and food efficiency with the inclusion of increasing levels of the meal. The fat content in milk showed a linear increasing effect. Ditto for cholesterol. It is recommended to include not more than 7.00% of licuri meal for lactating cows with an average production of 20 kg day-1.

Fermentation characteristics, chemical composition and fractionation of carbohydrates and crude protein of silage of elephant grass wilted or with addition of castor bean meal / Características fermentativas, composição química e fracionamento de carboidratos e proteínas de silagem de capim-elefante emurchecido ou com adição de torta de mamona

The experiment was conducted to evaluate the concentrations of ammonia nitrogen, hydrogen potential, the losses of deriving the fermentative process, nutritional value, the fractioning of carbohydrates and protein the elephant grass silage wilted or not containing castor bean meal. The experimental design was completely randomized, with five treatments and with four replications: elephant grass wilted; elephant not wilted; elephant grass more castor bean meal (6%); elephant grass more castor bean meal (12%) and elephant grass more castor bean meal (18%), the coproduct was added with base on natural matter. We adopted a specific mass of 600 kg/m3. The silage containing 18% castor bean meal showed higher (P 0.05) pH (4.8). High level of ammoniacal nitrogen was observed in the silage grass not wilted (12.8% N-total). The castor bean meal was effective in reducing (P 0.05) losses to by effluent and total, providing a higher rate of recovery of dry matter. Also, was efficient in increasing (P 0.05) the levels of DM, ash, crude protein e lignin. Already for organic matter, acid detergent insoluble nitrogen, total digestible nutrients, neutral detergent fiber, acid detergent fiber, cellulose and total carbohydrate decreased (P 0.05) with increasing doses of the castor bean meal. For the carbohydrate fractioning, there was no difference (P>0.05) among the silages with additives for fractions A+B1, B2 and C. Forthe protein fractioning, the fractions A and C decreased (P<0.05) with increase of the inclusion of castorbean meal, differently, of the fraction B1+B2 which increased. The castor bean stands out as a goodadditive in silage of elephant grass to reduce moisture and improve the fermentation characteristics ofsilages also was effective in increasing the protein value of silages, especially when using the dose 18%.(AU)

O experimento foi conduzido para avaliar os teores de nitrogênio amoniacal, potencial hidrogeniônico, perdas decorrentes do processo fermentativo, composição química, fracionamento de carboidratos e de proteínas de silagens de capim-elefante emurchecido, ou não, contendo torta de mamona. O delineamento experimental adotado foi o inteiramente casualizado, com cinco tratamentos e quatro repetições: capim-elefante emurchecido; capim-elefante não emurchecido; capim-elefante mais torta de mamona (6%); capim-elefante mais torta de mamona (12%) e capim-elefante mais torta de mamona (18%). o coproduto foi adicionado com base na matéria natural. Adotou-se uma massa específica de 600 kg/m3. A silagem contendo 18% de torta de mamona apresentou maior (P 0,05) valor de Ph (4,8). Teor elevado de nitrogênio amoniacal foi verificado para a silagem de capim não emurchecido (12,8% N-total). A torta de mamona foi eficiente em reduzir (P 0,05) as perdas por efluente e total, proporcionando maior índice de recuperação da matéria seca. Além disso, mostrou-se eficiente em aumentar (P 0,05) os teores de matéria seca, matéria mineral, proteína bruta e lignina. Já a matéria orgânica, nitrogênio insolúvel em detergente ácido, nutrientes digestíveis totais, fibra em detergente neutro, fibra em detergente ácido, celulose e carboidratos totais diminuíram (P 0,05) com a inclusão da torta de mamona. Para o fracionamento de carboidratos, não se verificou diferença (P>0,05) entre assilagens aditivadas para as frações A+B1, B2 e C. Para o fracionamento de proteína, as frações A e Cdiminuíram (P<0,05) com o aumento de inclusão da torta de mamona, diferentemente, da fração B1+B2que se elevou. A torta de mamona destaca-se como um bom aditivo na ensilagem de capim-elefante, porreduzir a umidade e melhorar o perfil fermentativo das silagens, além disso, foi eficiente em incrementaro teor de proteína bruta das silagens, principalmente quando se utiliza a dose de 18%.(AU)

Fermentation characteristics, chemical composition and fractionation of carbohydrates and crude protein of silage of elephant grass wilted or with addition of castor bean meal / Características fermentativas, composição química e fracionamento de carboidratos e proteínas de silagem de capim-elefante emurchecido ou com adição de torta de mamona

The experiment was conducted to evaluate the concentrations of ammonia nitrogen, hydrogen potential, the losses of deriving the fermentative process, nutritional value, the fractioning of carbohydrates and protein the elephant grass silage wilted or not containing castor bean meal. The experimental design was completely randomized, with five treatments and with four replications: elephant grass wilted; elephant not wilted; elephant grass more castor bean meal (6%); elephant grass more castor bean meal (12%) and elephant grass more castor bean meal (18%), the coproduct was added with base on natural matter. We adopted a specific mass of 600 kg/m3. The silage containing 18% castor bean meal showed higher (P 0.05) pH (4.8). High level of ammoniacal nitrogen was observed in the silage grass not wilted (12.8% N-total). The castor bean meal was effective in reducing (P 0.05) losses to by effluent and total, providing a higher rate of recovery of dry matter. Also, was efficient in increasing (P 0.05) the levels of DM, ash, crude protein e lignin. Already for organic matter, acid detergent insoluble nitrogen, total digestible nutrients, neutral detergent fiber, acid detergent fiber, cellulose and total carbohydrate decreased (P 0.05) with increasing doses of the castor bean meal. For the carbohydrate fractioning, there was no difference (P>0.05) among the silages with additives for fractions A+B1, B2 and C. Forthe protein fractioning, the fractions A and C decreased (P<0.05) with increase of the inclusion of castorbean meal, differently, of the fraction B1+B2 which increased. The castor bean stands out as a goodadditive in silage of elephant grass to reduce moisture and improve the fermentation characteristics ofsilages also was effective in increasing the protein value of silages, especially when using the dose 18%.

O experimento foi conduzido para avaliar os teores de nitrogênio amoniacal, potencial hidrogeniônico, perdas decorrentes do processo fermentativo, composição química, fracionamento de carboidratos e de proteínas de silagens de capim-elefante emurchecido, ou não, contendo torta de mamona. O delineamento experimental adotado foi o inteiramente casualizado, com cinco tratamentos e quatro repetições: capim-elefante emurchecido; capim-elefante não emurchecido; capim-elefante mais torta de mamona (6%); capim-elefante mais torta de mamona (12%) e capim-elefante mais torta de mamona (18%). o coproduto foi adicionado com base na matéria natural. Adotou-se uma massa específica de 600 kg/m3. A silagem contendo 18% de torta de mamona apresentou maior (P 0,05) valor de Ph (4,8). Teor elevado de nitrogênio amoniacal foi verificado para a silagem de capim não emurchecido (12,8% N-total). A torta de mamona foi eficiente em reduzir (P 0,05) as perdas por efluente e total, proporcionando maior índice de recuperação da matéria seca. Além disso, mostrou-se eficiente em aumentar (P 0,05) os teores de matéria seca, matéria mineral, proteína bruta e lignina. Já a matéria orgânica, nitrogênio insolúvel em detergente ácido, nutrientes digestíveis totais, fibra em detergente neutro, fibra em detergente ácido, celulose e carboidratos totais diminuíram (P 0,05) com a inclusão da torta de mamona. Para o fracionamento de carboidratos, não se verificou diferença (P>0,05) entre assilagens aditivadas para as frações A+B1, B2 e C. Para o fracionamento de proteína, as frações A e Cdiminuíram (P<0,05) com o aumento de inclusão da torta de mamona, diferentemente, da fração B1+B2que se elevou. A torta de mamona destaca-se como um bom aditivo na ensilagem de capim-elefante, porreduzir a umidade e melhorar o perfil fermentativo das silagens, além disso, foi eficiente em incrementaro teor de proteína bruta das silagens, principalmente quando se utiliza a dose de 18%.