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Ivermectin (IVM) is one of the most widely used antiparasitic drugs worldwide and has become the drug of choice for anthelmintic and tick treatment in beef cattle production. Drugs used in production animals requires a withdrawal period after treatment to avoid residual concentrations above the defined maximun residue level (MRL). The aims of this study were to quantify the residue level of IVM in different muscles of cattle at several different time periods following 1% or 3.15% IVM administration and also determined whether the residue concentration was affected by industrial thermal processing. Distinct pattern of IVM residue distribution was observed in each tissue evaluated. After completed the IVM withdrawal periods, nearly all tissues presented IVM residues concentration below the established Codex Alimentarius MRLs, except for the injection site, which means that the calculation to determine the withdrawal period for the injection site should not be the same as that used for standard edible tissues. Also, the thermal processing to which canned products are exposed causes changes in the levels of quantified residues in comparison to raw products, which must be taken into consideration in surveillance program assessments and generates concerns regarding the possibility of residue limit violation in importing markets., Competing Interests: Conflict of interestThis research article has no conflicts of interest., (© Association of Food Scientists & Technologists (India) 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

This study aimed to explore the differences in the lipidome and mitochondrial fraction metabolome of Nellore cattle meat in different ranges of ultimate pH (pHu) normal (≤5.79), intermediate (5.80 to 6.19) and high (≥ 6.20) after 3- and 21-d postmortem. Instrumental color, myoglobin redox state, oxygen consumption, and metmyoglobin-reducing activity were measured during storage. A total of 472 lipids and 22 mitochondrial fraction metabolites were identified. Beef with high pHu showed positive regulation of ceramides involved in apoptosis and negative regulation of lipid classes related to membrane permeability and stability. In addition, lower carnitine content was noted in high-pHu beef than in normal-pHu beef. Acylcarnitines, phosphatidylinositol, and IMP showed upregulation in beef with intermediate pHu, indicating changes mainly related to energy, purine and pyruvate metabolism. Aging time impacted on the lipid content and metabolites involved in different metabolic pathways. These results provided new insights into beef's mitochondrial fraction lipid and metabolic profile with different pHu. In addition, beef with intermediate pHu differs from beef with high pHu due to changes in energy metabolism., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Gait visual scores are widely applied to horse breeding because they are a fast and easy phenotyping strategy, allowing the numeric interpretation of a complex biological process such as gait quality. However, they may suffer from subjectivity or high environmental influence. We aimed to investigate potential causal relationships among six visual gait scores in Campolina horses. The data included 5475 horses with records for at least one of the following traits: Dissociation (Di), Comfort (C), Style (S), Regularity (R), Development (De), and Gait total Scores (GtS). The pedigree comprised three generations with 14,079 horses in the additive relationship matrix. Under a Bayesian framework, (co)variance components were estimated through a multitrait animal model (MTM). Then, the inductive causation algorithm (IC) was applied to the residual (co)variance matrix samples. The resulting undirected graph from IC was directed in 6 possible causal structures, each fitted by a structural equation model. The final causal structure was chosen based on deviance information criteria (DIC). It was found that S significantly impacts the causal network of gait, directly and indirectly affecting C. The indirect causal effect of S on C was through the direct effect of S on De, then the direct effect of De on R, and finally, the direct effect of R on C. Di was caused by S, which is the reason for the genetic correlation between Di and GtS, due to causal effects being added to the model, they absorb the genetic correlation between Di and GtS. Those paths have biological meaning to horse movements and can help breeders and researchers better understand horses' complex causal network of gait., (© 2024 The Author(s). Journal of Animal Breeding and Genetics published by John Wiley & Sons Ltd.)

The objective of this work was to evaluate the effect of using naturally shaded pastures on scrotal thermoregulatory capacity, testicular echotexture, and sperm morphology of Nelore (Bos indicus) and Canchim (5/8 Bos taurus x 3/8 Bos indicus) bulls in a tropical climate region. Sixty-four adult Nelore and Canchim bulls were used, equally allocated in Full Sun (FS, n = 32) or Crop-Livestock-Forestry (CLF, n = 32) pasture systems. During five consecutive climate seasons, the bulls underwent monthly breeding soundness evaluations and the biometeorological variables in the systems were continuously monitored. Microclimate was significantly different between systems. CLF system had lower BGHI than FS throughout the experimental period. No triple interaction (Season x Breed x Treatment, P > 0.05) was observed for any of the variables. Animals in CLF showed lower body temperature in Summer (FS:39.41 ± 0.05 vs. CLF:39.30 ± 0.05 °C; P = 0.005) and in Autumn (FS:39.54 ± 0.05 vs. CLF:39.35 ± 0.05 °C; P = 0.005). Access to shading did not determine differences in the evolution of scrotal biometry, temperatures, and scrotal thermal gradients (P > 0.05). Regardless of breed, animals in CLF showed greater right testicular volume (FS:247.5 ± 5.7 vs. CLF:259.0 ± 5.7 cm³; P < 0.05), more suitable parenchyma echotexture, and fewer microlithiasis spots in the Spring and Summer. Testosterone concentration was higher in FS (FS:2.6 ± 0.2 vs. CLF:2.1 ± 0.2 ng/mL; P = 0.035). Canchim bulls presented higher total sperm defects during the Autumn and Winter (P = 0.010), but the total defects levels for Canchim and Nelore bulls were in normal range for adult bulls. Thus, the natural shade in CLF system was effective in improving the microclimate of pastures and minimizing adverse environmental effects on some reproductive features of interest in beef cattle., (© 2024. The Author(s) under exclusive licence to International Society of Biometeorology.)

The objective of this exploratory study was to assess the changes on lipidome and metabolome profiling of Longissimus lumborum bull muscle with different ultimate pH (pHu) and aging periods. The bull muscles classified as normal, intermediate, or high pHu were collected from a Brazilian commercial slaughterhouse, cut into steaks, individually vacuum-packaged, and aged for 3 days (3-d) or 21 days (21-d) at 2 °C. Muscle extracts were analyzed for the profiles of both lipids, by mass spectrometry (via direct flow-injection), and metabolites, by nuclear magnetic resonance, with downstream multivariate data analysis. As major results, pairwise comparisons identified C12:0 and C14:0 acylcarnitines as potential biomarkers of the intermediate pHu-muscle, which are related to lipid catabolism for alternative energy metabolism and indicate less protein breakage postmortem. Interestingly, the concentration of arginine at early postmortem aging (3-d) may influence the previously reported improved tenderness in normal and high pHu-muscles. Moreover, upregulation of fumarate, formate, and acetate with increased pHu muscle at 21-d aging indicate more intense tricarboxylic acid cycle, amino acid degradation, and pyruvate oxidation by reactive oxygen species, respectively. These three compounds (fumarate, formate, and acetate) discriminated statistically the muscle with high pHu at 21-d aging. The normal pHu-muscle showed higher concentrations of glycogenolysis and glycolysis metabolites, including glucose, mannose, and pyruvate. Hence, our results enhance knowledge of postmortem biochemical changes of beef within different pHu groups aged up to 21 days, which is essential to understand the mechanisms underpinning bull meat quality changes., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Carmen Josefina Contreras Castillo reports financial support was provided by State of Sao Paulo Research Foundation. Alan Giovanini de Oliveira Sartori reports financial support was provided by State of Sao Paulo Research Foundation. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Bos taurus indicus bulls are very susceptible to pre-slaughter stress, which directly impacts the decline in muscle pH, leading to darker meat. The aim was to investigate the effect of succinate and atmosphere on the color stability of Nellore (Bos taurus indicus) Longissimus lumborum steaks classified by ultimate pH (pHu): normal pHu (5.40 ≤ pHu ≤ 5.79) and high pHu (pHu ≥ 5.80). The experimental treatment systems were: (i) vacuum packaging without succinate injection, (ii) HiOx-MAP (80 % O 2  + 20 % CO 2 ), and (iii) HiOx-MAP (80 % O 2  + 20 % CO 2 ) enhanced with sodium succinate injection (pH 5.4). Steaks from all treatment systems were stored at 4 °C for 14 days and tested for instrumental color, myoglobin content, oxygen consumption (OC), metmyoglobin-reducing activity (MRA), lipid oxidation, and microbiological analysis. High and normal pHu vacuum-packaged steaks exhibited greater color stability due to higher MRA. High and normal pHu steaks packaged with HiOx-MAP or HiOx-MAP enhanced with succinate showed improved color due to lower deoxymyoglobin content (%DMb) and OC up to the eighth day of storage. Still, succinate injection promoted increased (P < 0.05) lipid oxidation in normal pHu steaks and reduced MRA after 14 days. These findings emphasize the intricate interplay between pHu and packaging systems on Bos taurus indicus meat quality. Further research in this area could contribute to a better understanding of meat color abnormalities and provide insights into potential meat preservation and enhancement strategies., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Pork is of great importance in world trade and represents the largest source of fatty acids in the human diet. Lipid sources such as soybean oil (SOY), canola (CO), and fish oil (FO) are used in pig diets and influence blood parameters and the ratio of deposited fatty acids. In this study, the main objective was to evaluate changes in gene expression in porcine skeletal muscle tissue resulting from the dietary oil sources and to identify metabolic pathways and biological process networks through RNA-Seq. The addition of FO in the diet of pigs led to intramuscular lipid with a higher FA profile composition of C20:5 n-3, C22:6 n-3, and SFA (C16:0 and C18:0). Blood parameters for the FO group showed lower cholesterol and HDL content compared with CO and SOY groups. Skeletal muscle transcriptome analyses revealed 65 differentially expressed genes (DEG, FDR 10%) between CO vs SOY, and 32 DEG for CO vs FO, and 531 DEG for SOY vs FO comparison. Several genes, including AZGP1, PDE3B, APOE, PLIN1, and LIPS, were found to be down-regulated in the diet of the SOY group compared to the FO group. The enrichment analysis revealed DEG involved in lipid metabolism, metabolic diseases, and inflammation between the oil groups, with specific gene functions in each group and altered blood parameters. The results provide mechanisms to help us understand the behavior of genes according to fatty acids., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: The high similarity in anatomical and neurophysiological processes between pigs and humans make pigs an excellent model for metabolic diseases and neurological disorders. Lipids are essential for brain structure and function, and the polyunsaturated fatty acids (PUFA) have anti-inflammatory and positive effects against cognitive dysfunction in neurodegenerative diseases. Nutrigenomics studies involving pigs and fatty acids (FA) may help us in better understanding important biological processes. In this study, the main goal was to evaluate the effect of different levels of dietary soybean oil on the lipid profile and transcriptome in pigs' brain tissue., Results: Thirty-six male Large White pigs were used in a 98-day study using two experimental diets corn-soybean meal diet containing 1.5% soybean oil (SOY1.5) and corn-soybean meal diet containing 3.0% soybean oil (SOY3.0). No differences were found for the brain total lipid content and FA profile between the different levels of soybean oil. For differential expression analysis, using the DESeq2 statistical package, a total of 34 differentially expressed genes (DEG, FDR-corrected p-value < 0.05) were identified. Of these 34 DEG, 25 are known-genes, of which 11 were up-regulated (log2 fold change ranging from + 0.25 to + 2.93) and 14 were down-regulated (log2 fold change ranging from - 3.43 to -0.36) for the SOY1.5 group compared to SOY3.0. For the functional enrichment analysis performed using MetaCore with the 34 DEG, four pathway maps were identified (p-value < 0.05), related to the ALOX15B (log2 fold change - 1.489), CALB1 (log2 fold change - 3.431) and CAST (log2 fold change + 0.421) genes. A "calcium transport" network (p-value = 2.303e-2), related to the CAST and CALB1 genes, was also identified., Conclusion: The results found in this study contribute to understanding the pathways and networks associated with processes involved in intracellular calcium, lipid metabolism, and oxidative processes in the brain tissue. Moreover, these results may help a better comprehension of the modulating effects of soybean oil and its FA composition on processes and diseases affecting the brain tissue., (© 2023. The Author(s).)

The aim of this study was to identify the differentially expressed genes (DEG) from the skeletal muscle and liver samples of animal models for metabolic diseases in humans. To perform the study, the fatty acid (FA) profile and RNA sequencing (RNA-Seq) data of 35 samples of liver tissue (SOY1.5, n = 17 and SOY3.0, n = 18) and 36 samples of skeletal muscle (SOY1.5, n = 18 and SOY3.0, n = 18) of Large White pigs were analyzed. The FA profile of the tissues was modified by the diet, mainly those related to monounsaturated (MUFA) and polyunsaturated (PUFA) FA. The skeletal muscle transcriptome analysis revealed 45 DEG (FDR 10%), and the functional enrichment analysis identified network maps related to inflammation, immune processes, and pathways associated with oxidative stress, type 2 diabetes, and metabolic dysfunction. For the liver tissue, the transcriptome profile analysis revealed 281 DEG, which participate in network maps related to neurodegenerative diseases. With this nutrigenomics study, we verified that different levels of soybean oil in the pig diet, an animal model for metabolic diseases in humans, affected the transcriptome profile of skeletal muscle and liver tissue. These findings may help to better understand the biological mechanisms that can be modulated by the diet.

Background: Among the undesirable changes associated with obesity, one possibility recently raised is dysbiosis of the intestinal microbiota. Studies have shown changes in microbiota in obese rats and humans, but there are still few studies that characterize and compare the fecal microbiota of lean, obese and dogs after weight loss. Thus, this study aimed to evaluate the effects of a weight loss program (WLP) in fecal microbiota of dogs in addition to comparing them with those of lean dogs. Twenty female dogs of different breeds, aged between 1 and 9 years were selected. They were equally divided into two groups: Obese group (OG), with body condition score (BCS) 8 or 9/9, and body fat percentage greater than 30%, determined by the deuterium isotope dilution method, and lean group (LG) with BCS 5/9, and maximum body fat of 15%. Weight loss group (WLG) was composed by OG after loss of 20% of their current body weight. Fecal samples were collected from the three experimental groups. Total DNA was extracted from the feces and these were sequenced by the Illumina methodology. The observed abundances were evaluated using a generalized linear model, considering binomial distribution and using the logit link function in SAS (p < 0.05)., Results: The WLP modulated the microorganisms of the gastrointestinal tract, so that, WLG and LG had microbial composition with greater biodiversity than OG, and intestinal uniformity of the microbiota (Pielou's evenness index) was higher in OG than WLG dogs (P = 0.0493) and LG (P = 0.0101). In addition, WLG had values of relative frequency more similar to LG than to OG., Conclusion: The fecal microbiota of the studied groups differs from each other. The weight loss program can help to reverse the changes observed in obese dogs., (© 2022. The Author(s).)

This observational study aimed to evaluate serum and urinary amino acid (AA) concentrations in healthy dogs and dogs with chronic kidney disease (CKD) fed a commercial therapeutic renal diet with reduced protein and phosphorus levels. Ten dogs with CKD stages 3 or 4 composed the study group and received the renal diet for 180 days (RG T180). A control group (CG T30) composed of seven healthy dogs was fed a renal diet for 30 days. When comparing serum AA between RG T180 and CG T30, histidine, isoleucine, leucine, lysine, phenylalanine, tryptophan, cysteine, citrulline, ornithine, taurine, branched-chain amino acids (BCAA), and total essential amino acids (EAA) were higher in RG T180. Meanwhile, arginine, asparagine, aspartate, glutamine, serine, and tyrosine were higher in CG T30. Serum phenylalanine, tryptophan, and hydroxyproline were higher in RG T0 (dogs with CKD before consuming a renal diet) when compared to RG T180. In addition, the serum ratios of arginine/citrulline, tyrosine/phenylalanine, and serine/glycine were higher in CG T30 than in RG T180. Concerning urinary AA concentrations in CKD dogs, isoleucine, phenylalanine, tryptophan, aspartate, cysteine, and BCAA were higher in RG T180. In urine, the total EAA/total non-essential AA ratio in RG T180 was higher than in CG T30 as well as tyrosine/phenylalanine ratio higher in CG T30. In conclusion, the combination of renal diet and conservative treatment over 6 months in dogs with CKD stages 3 or 4 affected the AAs metabolism when compared to healthy adult dogs.

Background: Homemade diets are a worldwide trend, and many recipes are currently available on websites but may not be considered balanced. This study aimed to evaluate if the number of ingredients, supplement, or vegetarian/vegan-only ingredients included in a recipe influence the nutrient content of homemade diets for dogs and cats. Chemical analyses were performed on 75 diets for dogs and 25 for cats prepared according to websites' recipes, and minerals were analyzed by inductively coupled plasma optical emission spectrometry (ICP-OES)., Results: Canine diets that met fat requirements had more ingredients than those below recommendations. None of the cat diets met iron requirements, and feline diets that met requirements of manganese had fewer ingredients and those that supplied requirements of protein and sodium had more ingredients than the diets below recommendations (p < 0.05). Non-supplemented canine and feline diets had calcium and calcium:phosphorus ratio below recommendations (p < 0.05). Non-supplemented feline diets had lower sodium and iron, and zinc levels were below recommendation in diets for both species. Diets with animal products for dogs had higher levels of protein and zinc, although zinc was deficient in both groups, and there were higher amounts of crude fiber, magnesium, and manganese in vegetarian/vegan diets (p < 0.05). Diets with animal products for cats had higher levels of protein (p = 0.003), but there was a higher amount of crude fiber (p = 0.014) in vegetarian/vegan diets., Conclusion: The number of ingredients and vegetarian/vegan preparation did not guarantee nutritional adequacy of diets, and the presence of supplement did not ensure a balanced diet., (© 2021. The Author(s).)

Nowadays, there is a growing concern about contamination of toxic metals (TM) in pet food due to the great potential for health risks of these elements. TM concentrations in commercial pet foods (n = 100) as well as in ingredients used in their composition (n = 100) were analyzed and compared to the Food and Drug Administration (FDA) maximum tolerable level (MTL), and the TM concentrations found in the different sources of carbohydrate, protein, and fat were compared. The TM concentrations were determined by inductively coupled plasma with optical emission spectrometry (ICP-OES). Concentrations above the MTL for aluminum, mercury, lead, uranium, and vanadium were observed in both dog and cat foods, and the percentage of dog foods that exceeded the MTL of these TM were: 31.9%; 100%; 80.55%; 95.83%; and 75%, respectively, and in cat foods: 10.71%; 100%; 32.14%; 85.71%; 28.57%, respectively. The MTL values of these TMs and the mean values in dog foods (mg/kg dry matter basis) (MTL [mean ± standard deviation]) were: aluminum: 200 (269.17 ± 393.74); mercury: 0.27 (2.51 ± 1.31); lead: 10 (12.55 ± 4.30); uranium: 10 (76.82 ± 28.09); vanadium: 1 (1.35 ± 0.69), while in cat foods were: aluminum: 200 (135.51 ± 143.95); mercury: 0.27 (3.47 ± 4.31); lead: 10 (9.13 ± 5.42); uranium: 10 (49.83 ± 29.18); vanadium: 1 (0.81 ± 0.77). Dry foods presented higher concentrations of most TM (P < 0.05) than wet foods (P < 0.05). Among the carbohydrate sources, there were the highest levels of all TM except cobalt, mercury, and nickel in wheat bran (P < 0.05), while among the protein sources, in general, animal by-products had higher TM concentrations than plant-based ingredients. Pork fat had higher concentrations of arsenic, mercury, and antimony than fish oil and poultry fat. It was concluded that the pet foods evaluated in this study presented high concentrations of the following TM: aluminum, mercury, lead, uranium, and vanadium., (© 2021. The Author(s).)

Obesity is characterized by a low degree of chronic inflammation state that, along with metabolic modifications, promotes important changes in the animal's organism. Adipose tissue actively participates in inflammation and immunity, and several defense cells of the organism may, therefore, be involved in the diversity found between obese and ideal weight individuals. Studies regarding this subject have shown immune cell changes in humans and rats, however, the literature is scarce in relation to dogs. Thus, the present study aimed to evaluate the gene expression profile of immunoinflammatory response and the lymphoproliferation of obese dogs before and after weight loss. Eight female dogs, neutered, of different breeds, aged between 1 and 8 years (4.74±3.19), obese, with body condition score (BCS) of 9 out of a 9-point scale and body composition determined by the deuterium isotope dilution method were included. The obese dogs were enrolled in a weight loss program and after losing 20% of their initial weight became a second experimental group. A third experimental group consisted of eight female dogs, neutered, aged between 1 and 8 years (3.11±0.78) and with ideal BCS (5 out of a 9-point scale). Gene expression of immunoinflammatory cytokines (resistin, leptin, adiponectin, TNF-α, IL-6, IL-8, and IL-10) was assessed by qRT-PCR and immunity was assessed by lymphoproliferative response using the flow cytometry technique. The data that presented normal distribution was evaluated by analysis of variance by the PROC MIXED of the SAS and when differences were detected, these were compared by the Tukey test. Regarding the gene expression data, the procedure PROC GLIMMIX was adopted and the methodology of generalized linear model was used, in which the Gama distribution proved to be adequate. Values of p<0.05 were considered significant. The mean weight loss period of the animals included in the study was 194.25 ± 28.31 days and the mean weekly weight loss rate was 1.02 ± 0.82%. The average fat mass, both in percentage (P<0.001) and in kilograms (P = 0.012), was higher in the obese group (40.88%; 8.91kg), returning to normal and without difference between the control group (19.16%; 3.01kg) and after weight loss (22.10%; 4.11kg). The weight loss program resulted in an increase in percentage of lean body mass (P = 0.001), 55.50% in obese animals vs 77.90% in obese dogs after weight loss, the latter with no difference when compared to the control group (80.84%). The obese group presented increased gene expression of resistin and IL-8 in relation to the weight loss group (P = 0.002). In adiponectin, the obese group presented increased mRNA gene expression when compared to the weight loss group (P = 0.003). The evaluation of lymphocyte proliferation showed differences between the group of obese animals before and after weight loss (P = 0.004). Weight loss resulted in an increase in the lymphoproliferation rate (18.48%) compared to obese dogs at the beginning of the study (10.71%). These results indicate that weight loss modulates the immunoinflammatory response of obese dogs and may present important benefits to health and longevity of dogs., Competing Interests: The authors have read the journal’s policy and have the following potential competing interests: R.P. and C.R.F.P. are paid employees of Grandfood Indústria e Comércio. This does not alter our adherence to PLOS ONE policies on sharing data and materials. There are no patents, products in development or marketed products associated with this research to declare.

The potential of dams as oocyte donors can be a selection criterion for animal breeding programs, but also an involuntary driver of the process. In both cases, it is important to determine genetic components influencing the outcome of in vitro embryo production (IVEP). The objective of the present study was to perform a detailed genetic analysis for in vitro embryo production traits in Dairy Gir cows. A dataset containing 11,450 records of ovum pick-up (OPU) and in vitro fertilization (IVF) procedures from 2684 Dairy Gir donors was evaluated. Analyzed traits were number (N OV ) and percentage (P OV ) of viable oocytes; number (N GI ) and percentage (P GI ) of grade I oocytes; number (N EMB ) and percentage (P EMB ) of viable embryos. All analyzes were performed using animal models by a Bayesian framework. Heritability estimates varied from 0.16 to 0.32 for count traits and from 0.01 to 0.06 for percentage traits. The proportion of the total variation represented by the additive genetic effect of sire (semen used in IVF) for N EMB and P EMB was 7% and 5% respectively. Associations between estimated breeding values from progeny tested bulls for IVEP traits, milk production, age at first calving and conformation traits were mainly low or close to zero. Results indicate that selection for IVEP traits is possible in Dairy Gir cattle and would not impair genetic progress for traits already considered as selection criteria. The N OV seems to be a promising target trait. However, a selection index could help to avoid the use of sires with negative genetic merit for percentage traits, minimizing possible deterioration in the long term., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest to this publication., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Pet owners often don't acknowledge the need for home-prepared diet formulation by a trained professional and may use recipes from sources such as the internet. Macronutrient and mineral composition of home-prepared diets were analyzed and compared to NRC and FEDIAF recommendations, and heavy metal concentrations were analyzed and compared to FDA maximum tolerable levels (MTL) for dogs and cats. Recipes of home-prepared diets for adult dogs (n = 75) and cats (n = 25) were evaluated. Analyses of protein, fat, and fiber were performed according to AOAC, and mineral and heavy metal analyses were performed using inductively coupled plasma optical emission spectrometry (ICP-OES). None of the diets supplied recommended levels of all nutrients evaluated, and more than 84.0% of diets presented three or more nutrients below recommendations. Nutrients with most levels below recommendations were calcium and potassium in recipes for dogs and iron and zinc in recipes for cats. As for heavy metals, levels of lead, cobalt, mercury, uranium, and vanadium were above MTLs. Results suggest that home-prepared diets may be a health risk to dogs and cats if not properly formulated. Furthermore, the chronic heavy metal intake must be better elucidated in order to understand the full impact of results.

The study was conducted with 40 lambs Merino × Texel in order to determine the effects of 10 or 30 mg of Cu/kg DM of copper sulphate and copper-methionine on the lipid and cholesterol metabolism. The lambs were randomly distributed into 5 treatments, with 8 animals each. The treatments were: control, without cooper additions; 10 or 30 mg of Cu/kg DM in the form of copper sulphate; 10 or 30 mg of Cu/kg DM in the form of copper-methionine. The hepatic concentration of copper was higher in lambs supplemented with copper. Total cholesterol, HDL cholesterol, LDL cholesterol and triglycerides in the serum, as well as the SFTU did not differ between treatments. The supplementation of copper regardless of source or level reduced the concentration of cholesterol in the LT muscle, and reduced the concentration of GSH and increased the concentration of GSSG in the liver. Additionally, there was a minimal effect on the fatty acid profile of lipid in lambs., (Copyright © 2019. Published by Elsevier Ltd.)

Our aim was to evaluate the effects of three thermal environments over time on kinetics, functionality and in vitro fertility of cryopreserved bovine spermatozoa. Four ejaculates from five bulls (n = 20) were cryopreserved. After thawing, semen was evaluated (0 hr), incubated for 4 hr in T36.0 (36.0°C), T38.0 (38.0°C) and T39.5 (39.5°C), and analysed every hour (1 hr, 2 hr, 3 hr, 4 hr). In vitro production of embryos was performed at 0 hr and 4 hr. Sperm motility and cell kinetics (Computer-Assisted Sperm Analysis) were impaired after 2 hr at T38.0 and T39.5 (p < 0.05). Flow cytometry revealed an increase in the cells with injured plasma membrane to 39.5°C and a general reduction in the mitochondrial potential over time (p < 0.05). In vitro fertility was impaired in all temperatures after 4 hr, but there was no difference between 36.0°C and 38.0°C. Our results suggest that the ex situ resilience of semen at 36.0°C after thawing with no major damage to the quality is limited to 3 hr. In normothermia or in thermal stress, sperm cells present a gradual reduction of movement and functionality, which were more significant after 1 hr of incubation. The in vitro production of embryos is impaired when the semen is kept in a thermal environment ≥36.0°C for 4 hr., (© 2019 Blackwell Verlag GmbH.)

The purpose of this research was to investigate the causes and consequences of pHu variations in beef cattle. A group of 176 Nellore beef cattle was evaluated and classified into two different pHu groups: High (≥6.0, N = 17) and Normal (<5.8, N = 159). Plasma concentrations of cortisol and adrenocorticotropic hormone, lactate and glycogen muscular content, meat color, shear force and Longissimus thoracis muscle proteomic profile were evaluated and compared between pHu groups. Muscle glycogen content, meat color and shear force statistically differed between pHu groups. Label-free quantitative proteomic analysis revealed ten differentially abundant proteins between pHu groups, involved in metabolic processes and muscle contraction, which also were significantly correlated with pHu. Thirty-six and 31 proteins were exclusively present in Normal and High pHu group, respectively, which were related to TCA cycle, cortisol production, calcium regulation, and antioxidant function. The MYH7, UGP2, H2AFJ and VDAC3 were identified as potential indicators of pHu variations. CALM and NNT appeared to be interesting proteins to understand the metabolic pathways behind pHu. Data are available via ProteomeXchange with identifier PXD009320., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

In cattle, the oviduct plays a major role in the reproductive process; however, molecular control of oviduct receptivity to the embryo is poorly understood. A model for receptivity based on size of the pre-ovulatory follicle (POF) was used to compare oviductal morphology, cellular proliferation, and candidate transcript abundance. Growth of the POF of Nelore (Bos indicus) cows was manipulated to produce two groups: a large POF-large corpus luteum (CL) group (LF-LCL; greater receptivity) and a small POF-small CL group (SF-SCL). Samples of the ampulla and isthmus ipsilateral and contralateral to CL were collected 4 days after GnRH-induced ovulation. Tissues were either embedded in paraffin for Harris-Hematoxylin and Eosin and periodic acid-Schiff staining and KI67 immunostaining, followed by morphological analyses, or stored at -80 °C for RNA extraction, cDNA synthesis, and qPCR analyses. The effects of group (LF-LCL and SF-SCL), region (ampulla and isthmus), and side (ipsilateral and contralateral) were analyzed using three-way nested ANOVA. The ipsilateral ampulla of the LF-LCL group presented more primary mucosal folds, a greater mucosal-folding grade and luminal perimeter, and more secretory cells and proliferating cells when compared with the ampulla of the SF-SCL group and with the contralateral ampulla of both groups. There were no morphological differences in the isthmus between groups and sides. Changes in transcript abundance are suggestive of LF-LCL-stimulated secretory activity. In summary, ovulation of a larger POF generates a periovulatory endocrine milieu that modulates morphological and functional features of the bovine oviduct which may support embryo survival and development.

The objectives of this study were to characterize the allelic and genotypic frequencies of polymorphisms in the µ-calpain and calpastatin genes, and to assess their association with meat tenderness and animal growth in Nellore cattle. We evaluated 605 Nellore animals at 24 months of age, on average, at slaughter. The polymorphisms were determined for the molecular markers CAPN316, CAPN530, CAPN4751, CAPN4753, and UOGACAST1. Analyses of meat tenderness at 7, 14, and 21 days of maturation were performed in samples of longissimus thoracis obtained between the 12th and 13th rib and sheared using a Warner Bratzler Shear Force. Significant effects were observed for meat tenderness at days 7, 14, and 21 of maturation for the marker CAPN4751, at day 21 for the marker CAPN4753, and at days 14 and 21 for the marker UOGCAST1. For genotypic combinations of markers, the results were significant for the combination CAPN4751/UOGCAST1 in the three maturation periods and CAPN4753/UOGCAST1 at days 14 and 21 of maturation.

Melatonin may have beneficial effects when used in oocyte maturation and embryo development culture. The effect of melatonin during IVM on meiosis resumption and progression in bovine oocytes and on expression of antioxidant enzymes, nuclear fragmentation and free radicals, as well as on embryo development were assessed. Cumulus-oocyte complexes were matured in vitro with melatonin (10(-9) and 10(-6) M), FSH (positive control), or without hormones (negative control) in defined medium. Maturation rates were evaluated at 6, 12, 18, and 24 hours. Transcripts for antioxidant enzymes (CuZnSOD, MnSOD, and glutathione peroxidase 4 (GPX4)) in oocytes and cumulus cells, nuclear fragmentation in cumulus cells (TUNEL) and reactive oxygen species levels in oocytes (carboxy-H2 difluorofluorescein diacetate) were determined at 24 hours IVM. Effect of treatments on embryo development was determined after in vitro fertilization and culture. At 12 hours, meiosis resumption rates in FSH and melatonin-treated groups were similar (69.6%-81.8%, P > 0.05). At 24 hours, most oocytes were in metaphase II, with FSH showing highest rates (90.0%, P < 0.05) compared with the other groups (51.6%-69.1%, P > 0.05). In cumulus cells, MnSOD expression was higher in FSH group (P < 0.05) whereas Cu,ZnSOD transcripts were more abundant in melatonin group (10(-6)M; P < 0.05). Nuclear fragmentation in cumulus cells was highest in controls (37.4%/10,000 cells; P < 0.05) and lower in FSH and 10(-6)M melatonin (29.4% and 25.6%/10,000 cells, respectively). Reactive oxygen species levels were lower in oocytes matured with 10(-6)M melatonin than in control and FSH groups (P < 0.05). Embryo development from oocytes matured only with melatonin was similar to those matured in complete medium (P > 0.05). In conclusion, although melatonin during IVM in a defined medium does not stimulate nuclear maturation progression it does stimulate meiosis resumption and such treated oocytes support subsequent embryo development. Melatonin also shows cytoprotective effects on cumulus-oocyte complexes., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Purpose: Leber hereditary optic neuropathy (LHON) is a mitochondrial disease characterized by bilateral vision loss. More than 95% of LHON cases are associated with one of the three main mtDNA mutations: G11778A, T14484C, and G3460A. The other 5% of cases are due to other rare mutations related to the disease. The aim of this study was to identify the prevalence and spectrum of LHON mtDNA mutations, including the haplogroup, in a cohort of Brazilian patients with optic neuropathy and to evaluate the usefulness of iPLEX Gold/matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) technology in detecting LHON mutations., Methods: We analyzed a total of 101 patients; 67 had a clinical diagnosis of LHON and 34 had optic neuropathy of unknown etiology. Direct sequencing and iPLEX Gold/MALDI-TOF MS were used to screen for the most common pathogenic point mutations in LHON, together with the rare mutations G3733A, C4171A, T10663C, G14459A, C14482G, A14495G, C14568T, and C14482A., Results: We identified mutations in 36 patients, of whom 83.3% carried the G11778A mutation and 16.7% carried the T14484C mutation. In individuals with mutations, the haplogroups found were L1/L2, L3, C, R, U, D, and H. Rare mutations were not detected in any of the patients analyzed., Conclusions: The frequencies of the main LHON mutations were similar to those previously reported for Latin America. A different frequency was found only for the A3460G mutation. The most frequent haplogroups identified were of African origin. The iPLEX Gold/MALDI-TOF MS technology proved to be highly accurate and efficient for screening mutations and identifying the haplogroups related to LHON. The MassArray platform, combined with other techniques, enabled definitive diagnosis of LHON in 36% (36/101) of the cases studied.

Productivity rates directly depend on the fertility of a herd, which in turn can be influenced by many factors. Semen deposited in the female reproductive tract is foreign to the body and, in response to this invasion, produces an inflammatory reaction, which is characterized by rapid infusion of polymorphonuclear (PMN) cells. Techniques to obtain an endometrial sample are usually invasive and can mask the true inflammatory response. Ultrasound is a noninvasive technique and can contribute to the diagnosis of postartificial insemination (AI) inflammatory response in cattle. The present study was divided into two experiments. The aim of experiment 1 was to compare two methods of endometrial cytology collection, uterine cytobrush (UC) and uterine lavage (UL), and their effects on uterine hemodynamics that provide information about blood flow. The two methods were evaluated by Doppler ultrasound using the spectral and color modes. For that purpose, 19 Nellore cows were synchronized for timed AI and subjected to UC (n = 9) or UL (n = 10). The techniques were performed 4 hours after AI. The results showed that both techniques allow collection of a good quality sample and with enough PMN cells to perform counting. More PMN cells were obtained by UL than UC. There was no difference in uterine blood flow between the UC and UL groups in any of the periods evaluated (34 hours before and 4, 24, and 48 hours after collection of uterine sample). On the basis of results of experiment 1, the effect of UL on fertility was studied in experiment 2. A total of 128 Nellore cows were synchronized for TAI; 35 cows were subjected to endometrial cytology by UL 4 hours after AI, and 93 were not submitted to any procedure (control). Pregnancy diagnosis was performed by transrectal ultrasound 30 days after AI. Pregnancy rates did not differ between UL (54.29%) and control (56.99%) groups. The results of this study showed that UL allows the collection of more representative cells of the surface of the uterus than UC technique and causes no damage to the reproductive tract. Moreover, UL did not affect pregnancy rate when performed 4 hours after AI., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The objective of this study was to estimate variance components for oocyte and embryo production traits in Guzerá breed female donors, and investigate their associations with age at first calving (AFC). The traits analyzed were the number of viable oocytes (NOV), the number of grade I oocytes (NGI), the number of cleaved embryos (NCLV), and viable embryos produced (NEMB), and the percentages of viable oocytes (POV), grade I oocytes (PGI), cleaved embryos (PCLV), and viable embryos (PEMB). Data were obtained from 5173 ovary puncture and in vitro fertilization (IVF) sessions using 1080 Guzerá female donors of different ages, occurred from March 2005 to July 2013. Variables were log-transformed (logeX+1) prior to analysis. (Co)variance components were estimated by restricted maximum likelihood (REML), using one- and two-trait animal models. Permanent environment and IVF sire (father of the embryos) random effects were included. Estimated heritabilities for NOV, NGI, NCLV, NEMB, POV, PGI, PCLV, and PEMB were 0.19, 0.08, 0.16, 0.14, 0.04, 0.03, 0.01, and 0.07, respectively. Repeatabilities for count traits (NOV, NGI, NCLV, and NEMB) varied from 0.14 and 0.32, higher than estimated for percentage traits (POV, PGI, PCLV, and PEMB), which varied from 0.01 to 0.08. Selection for NOV may be more appropriate in breeding programs than selection for NEMB, because of its strong genetic correlation (0.68) with NEMB and its greater time- and cost-effectiveness. AFC was only weakly associated with the oocyte and embryo production traits, which indicates that there would be no effect on AFC when selecting for these traits.

The glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) are key components in the regulation of the hypothalamic-pituitary-adrenal neuroendocrine axis and coordinate the physiological response to stress agents to reestablish homeostasis. Genetic variations of GR (NR3C1) and MR (NR3C2) genes could explain the alterations in animals to adapt to challenges, and therefore, their influence on production traits. The present study aimed to identify single-nucleotide polymorphisms (SNPs) in the bovine NR3C1 and NR3C2 genes and explore their associations to relevant traits of beef cattle production. Genotypes and phenotypes were collected from 241 male Nellore cattle (119 noncastrated and 122 castrated surgically) with an average of 24 ± 1.2 mo of age and live weight of 508 ± 39 kg. The traits evaluated were concentrations of plasma adrenocorticotropic hormone (ACTH) and cortisol, muscle glycogen and lactate content, and pH, color, cooking loss, and shear force of longissimus thoracis measured on the 1st, 7th, and 14th days postmortem. Five SNPs were identified, 2 in the NR3C1 gene and 3 in the NR3C2 gene. There was an associative relationship between the SNP NR3C1&#95;1 g.3293A>G and postmortem plasma concentration of cortisol (P = 0.0008). The SNPs NR3C2&#95;1 g.115T>C and NR3C2&#95;2 g.570T>C were associated with muscle glycogen content (P = 0.0306 and P = 0.0158), postmortem plasma concentration of ACTH (P = 0.0118 and P = 0.0095), and cooking loss of the steak aged 1 d (P = 0.0398 and P = 0.0423). Haplotype analysis showed associations of GR haplotypes with postmortem plasma concentrations of cortisol and MR haplotypes with meat color, cooking losses, muscle glycogen content, and plasma concentrations of ACTH. The associations observed in the present study show that SNPs in GR and MR genes are related with changes of hypothalamic-pituitary-adrenal axis activity and metabolic profile in cattle, leading to individual variation in meat quality traits., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The objectives of this study were to characterize and define homogenous production environments of composite beef cattle in Brazil in terms of climatic and geographic variables by using multivariate exploratory techniques; to evaluate the presence of genotype by environment interaction (GxE) for post-weaning weight gain (PWG), yearling scrotal circumference (SC), and yearling muscling (MUS). Hierarchical and nonhierarchical cluster analysis was used to group farms located in regions with similar environmental variables into clusters. Six clusters of farms were formed. The effect of sire-cluster interaction was tested by single-trait analysis. Genetic parameters were estimated by multi-trait analysis considering the same trait to be different in each cluster. The effect of sire-cluster interaction was significant (P <0.01) for PWG and MUS. Estimates of genetic correlations among clusters ranged from 0.31 to 0.93 for PWG, 0.64 to 0.89 for SC, and 0.18 to 0.80 for MUS. These results indicate the need for a genetic analysis on a regional basis or inclusion of the GxE effect in the statistical model to permit appropriate evaluation of the animals.

Acute renal failure (ARF) is a common renal disease that can lead to high mortality. Recovery from ARF occurs with the replacement of necrotic tubular cells by functional tubular epithelial cells and the normalization of microvascular endothelial cell function in the peritubular capillaries. Conventional therapeutic techniques are often ineffective against ARF. Hence, stem cell therapies, which act through multiple trophic and regenerative mechanisms, are encouraging. We investigated the homing of human immature dental pulp stem cells (IDPSCs) after endovenous (EV) or intraperitoneal (IP) injection, in immunocompetent Wistar rats with ARF induced by intramuscular injection of glycerol, without the use of immunosuppression. The cells, which had been cryopreserved for 6 years, were CD105(+), CD73(+), CD44(+), and partly, STRO-1(+) and CD146(+), and presented unaltered mesoderm differentiation potential. The presence of these cells in the tubular region of the kidney and in the peritubular capillaries was demonstrated. These cells accelerate tubular epithelial cell regeneration through significant increase of Ki-67-immunoreactive cells in damaged kidney. Flow cytometry analysis confirmed that IDPSCs home to the kidneys (EV 34.10% and IP 33.25%); a lower percentage of cells was found in the liver (EV 19.05% and IP 9.10%), in the muscles (EV 6.30% and IP 1.35%), and in the lungs (EV 2.0% and IP 1.85%). After infusion into rat, these cells express pericyte markers, such as CD146(+), STRO-1(+), and vascular endothelial growth factor (VEGF(+)). We found that IDPSCs demonstrate renotropic and pericyte-like properties and contributed to restore renal tubule structure in an experimental rat ARF model.

Nellore beef cattle, a Bos indicus (Zebu) breed, is well adapted to tropical conditions and has allowed Brazil to become one of the largest producers of red meat. Nevertheless, B. indicus breeds are reported to have less tender meat than Bos taurus. This study was designed to identify genes associated with meat tenderness and thus provides important information for breeding programs. A group of 138 animals was evaluated for longissimus thoracis muscle shear force (SF). Animals with the highest and lowest SF values (six animals each) were then selected for protein abundance studies. Samples were subjected to two-dimensional gel electrophoresis (2-DE) followed by peptide sequencing through mass spectrometry (MS) to identify differentially expressed proteins associated with SF values. Seventeen differentially expressed spots were observed (p<0.05) between the two groups. The 13 proteins identified included structural proteins (alpha actin-1, MLC1, MLC3, MLC2F and tropomyosin), related to cell organization (HSPB1 and HSP70), metabolism (beta-LG, ACBD6 and Complex III subunit I) and some uncharacterized proteins. Results confirm the existence of differentially expressed proteins associated with SF, which can lead to a better understanding of mechanisms involved in meat tenderness., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The aim of the present study was to evaluate the effect of different modified atmosphere packaging (MAP) systems (vacuum, 75% O2+25% CO2 and 100% CO2) on the stability of lamb loins stored at 1±1°C for 28 days. Microbiological (counts of aerobic and anaerobic psychrotrophic microorganisms, coliform at 45°C, coagulase-positive staphylococci and lactic acid bacteria and presence of Salmonella), physical and chemical (thiobarbituric acid reactive substances [TBARS], objective color, pH, water loss from cooking [WLC] and shear force), sensory (acceptance testing using a 9-point hedonic scale) and gas composition analyses were performed. Lamb meat remained stable with respect to the majority of the evaluated physical and chemical indexes and within the standards established by Brazilian legislation for pathogenic microorganisms throughout the storage period in all three packaging systems. However, with respect to psychrotrophic microorganisms, 100% CO2 packaging system provided increased stability despite presenting lower appearance preference., (© 2013.)

Meat quality is an important feature for the poultry industry and is associated with consumer satisfaction. The calpain 1 (CAPN1) gene is related to the tenderness process of meat post- mortem, and the calpain 3 (CAPN3) gene plays an important role in myofibrillar organization and growth. The objective of the present study was to identify polymorphisms in these genes and to determine the association between these polymorphisms and traits of economic interest in poultry. Eleven animals (F₁) from an experimental poultry population at Embrapa Swine and Poultry were used to identify the polymorphisms. Four single nucleotide polymorphisms (SNPs) were found in the CAPN1 gene, and one SNP was found in the CAPN3 gene. A polymorphism from each gene was selected for genotyping in 152 chickens from the Embrapa F₂ experimental population and 311 chickens from a commercial population. Polymorphism g.2554T>C (CAPN1) was associated with body weight at 35 to 42 days, thigh weight, breast weight, carcass weight, and meat lightness content. SNP g.15486C>T (CAPN3) was associated with thigh yield, thawing-cooking loss, and shear force. Results suggest the possibility of using molecular markers in CAPN1 and CAPN3 genes as a tool for performance and meat quality traits in poultry breeding programs.

Ractopamine hydrochloride (RAC) improves swine production efficiency by redirecting nutrients to favor muscle accretion rather than fat deposition. In the present study, the time-dependent effect of RAC feeding on performance, plasma urea N (PUN) concentrations, and carcass traits of finishing pigs were evaluated. In a 28-d growth study, 80 barrows (average initial BW = 69.4 ± 7.9 kg) were assigned to 1 of 5 treatments in a randomized complete block design with 8 replicate pens per treatment and 2 pigs per pen. The pigs were fed a corn-soybean meal-based diet with no added RAC (control) or 10 mg of RAC/kg fed for 7, 14, 21, or 28 d before slaughter. All diets were formulated to contain 0.88% standardized ileal digestible Lys (1.0% total Lys) and 3.23 Mcal of ME/kg. Individual pig BW and pen feed disappearance were recorded weekly to determine BW changes, ADG, ADFI, and G:F. Anterior vena cava blood samples were taken on d 28 for determination of PUN concentrations. After 28 d on trial, the pigs were slaughtered and carcass measurements made at 24 h postmortem. Overall, providing pigs with different RAC feeding durations did not affect the final BW and ADFI but resulted in a tendency (P = 0.09) for a linear increase in ADG and a linear improvement (P = 0.003) in G:F. No effect of RAC feeding was found for weekly ADFI. Weekly improvements (P < 0.05) in ADG and G:F were observed over the first 21 d of RAC feeding. However, the growth response declined (P < 0.05) in wk 4 of RAC treatment. The concentrations of PUN exhibited a quadratic decrease (P = 0.004) as the RAC feeding duration increased. Although RAC feeding did not affect any backfat measurements and carcass length, increasing the RAC feeding duration linearly increased HCW (P = 0.01), dressing percentage (P = 0.03), LM depth (P = 0.001), LM area (P < 0.001), muscle-to-fat ratio (P = 0.004), and predicted carcass lean percentage (P = 0.02). These results indicate that a greater growth rate was achieved within the first 21 d of RAC feeding whereas the magnitude of carcass response was directly dependent on the duration of RAC feeding.

In the past, the focus of broiler breeding programs on yield and carcass traits improvement led to problems related to meat quality. Awareness of public concern for quality resulted in inclusion of meat quality traits in the evaluation process. Nevertheless, few genes associated with meat quality attributes are known. Previous studies mapped quantitative trait loci for weight at 35 and 42 days in a region of GGA4 flanked by the microsatellite markers, MCW0240 and LEI0063. In this region, there are 2 fibroblast growth factor binding protein (FGFBP) genes that play an important role in embryogenesis, cellular differentiation, and proliferation in chickens. The objective of this study was to identify and associate single nucleotide polymorphisms (SNPs) in FGFBP1 and FGFBP2 with performance, carcass, and meat quality in experimental and commercial chicken populations. In the commercial population, SNP g.2014G>A in FGFBP1 was associated with decreased carcass weight (P < 0.05), and SNP g.651G>A in FGFBP2 was associated with thawing loss and meat redness content (P < 0.05). Four haplotypes were constructed based on 2 SNPs and were associated with breast weight, thawing loss, and meat redness content. The diplotypes were associated with thawing loss, lightness, and redness content. The SNPs evaluated in the present study may be used as markers in poultry breeding programs to aid in improving growth and meat quality traits.

Given the important role of leptin in metabolism, we looked for a possible association of leptin and leptin receptor polymorphisms with carcass and growth traits in Nellore cattle. We examined associations of leptin and leptin receptor SNPs with ultrasound carcass (longissimus dorsi muscle area (ribeye area), backfat thickness and rump fat thickness and growth traits (weaning weight adjusted to 210 days of age, yearling weight adjusted to 550 days of age, weight gain of weaning to yearling and scrotal circumference adjusted to 550 days of age) of 2162 Bos primigenius indicus (Nellore) animals. Allele and genotypic frequencies were calculated for each marker. Allele substitution, additive and dominance effects of the polymorphisms were also evaluated. Some alleles of the molecular markers had low frequencies, lower than 1%, in the sample analyzed, although the same polymorphisms described for B. p. taurus cattle were found. Due to very low allelic frequencies, the E2JW, A59V and UASMS2 markers were not included in the analysis, because they were almost fixed. E2FB was found to be significantly associated with weight gain, ribeye area and backfat thickness. The promoter region markers, C963T and UASMS1, were also found to be significantly associated with ribeye area. T945M was significantly associated with weight gain. We conclude that the leptin and receptor gene markers would be useful for marker-assisted selection.

The embryonic developmental block occurs at the 8-cell stage in cattle and is characterized by a lengthening of the cell cycle and an increased number of embryos that stop development. The maternal-embryonic transition arises at the same stage resulting in the transcription of many genes. Gene expression studies during this stage may contribute to the understanding of the physiological mechanisms involved in the maternal-embryonic transition. Herein we identified genes differentially expressed between embryos with high or low developmental competence to reach the blastocyst stage using differential display PCR. Embryos were analysed according to developmental kinetics: fast cleavage embryos showing 8 cells at 48 h post insemination (hpi) with high potential of development (F8), and embryos with slow cleavage presenting 4 cells at 48 hpi (S4) and 8 cells at 90 hpi (S8), both with reduced rates of development to blastocyst. The fluorescence DDPCR method was applied and allowed the recovery of 176 differentially expressed bands with similar proportion between high and low development potential groups (52% to F8 and 48% in S4 and S8 groups). A total of 27 isolated fragments were cloned and sequenced, confirming the expected primer sequences and allowing the identification of 27 gene transcripts. PI3KCA and ITM2B were chosen for relative quantification of mRNA using real-time PCR and showed a kinetic and a time-related pattern of expression respectively. The observed results suggest the existence of two different embryonic genome activation mechanisms: fast-developing embryos activate genes related to embryonic development, and slow-developing embryos activate genes related to cellular survival and/or death.

Our objective was to estimate Bos primigenius taurus introgression in American Zebu cattle. One hundred and four American Zebu (Nellore) cattle were submitted to mtDNA, microsatellite and satellite analysis. Twenty-three alleles were detected in microsatellite analysis, averaging 4.6 ± 1.82/locus. Variance component comparisons of microsatellite allele sizes allowed the construction of two clusters separating taurus and indicus. No significant variation was observed when indicus and taurus mtDNA were compared. Three possible genotypes of 1711b satellite DNA were identified. All European animals showed the same restriction pattern, suggesting a Zebu-specific restriction pattern. The frequencies of B. primigenius indicus-specific microsatellite alleles and 1711b satellite DNA restriction patterns lead to an estimate of 14% taurine contribution in purebred Nellore.

The effect of melatonin during in vitro maturation (IVM) on DNA damage of cumulus cells (CCs) from bovine cumulus-oocyte complexes (COCs) and embryo development was evaluated. COCs from abattoir ovaries were cultured in maturation medium (MM) with 0.5μg/ml FSH and 5.0μg/ml LH (FSH-LH); 10(-9)M melatonin (MEL) or FSH-LH+MEL (FSH-LH-MEL). After 24h of in vitro maturation, the CCs surrounding the oocyte were subjected to DNA analysis by Comet assay. After in vitro fertilization and in vitro embryo culture, the embryo development rates were evaluated on day 2 post insemination (cleavage) and days 7-8 (blastocyst). The percentage of CCs with no DNA damage was significantly superior in MEL group (37.6±2.4) than in FSH-LH-MEL (28.0±2.4) and FSH-LH (17.8±2.41) groups. Cleavage and blastocysts rates were similar among groups. Melatonin during IVM protects the CCs from DNA damage but this effect did not influence embryo development in vitro., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Recently, superior cervical ganglionectomy has been performed to investigate a variety of scientific topics from regulation of intraocular pressure to suppression of lingual tumour growth. Despite these recent advances in our understanding of the functional mechanisms underlying superior cervical ganglion (SCG) growth and development after surgical ablation, there still exists a need for information concerning the quantitative nature of the relationships between the removed SCG and its remaining contralateral ganglion and between the remaining SCG and its modified innervation territory. To this end, using design-based stereological methods, we have investigated the structural changes induced by unilateral ganglionectomy in sheep at three distinct timepoints (2, 7 and 12 weeks) after surgery. The effects of time, and lateral (left-right) differences, were examined by two-way analyses of variance and paired t-tests. Following removal of the left SCG, the main findings were: (i) the remaining right SCG was bigger at shorter survival times, i.e. 74% at 2 weeks, 55% at 7 weeks and no increase by 12 weeks, (ii) by 7 weeks after surgery, the right SCG contained fewer neurons (no decrease at 2 weeks, 6% fewer by 7 weeks and 17% fewer by 12 weeks) and (iii) by 7 weeks, right SCG neurons were also larger and the magnitude of this increase grew substantially with time (no rise at 2 weeks, 77% by 7 weeks and 215% by 12 weeks). Interaction effects between time and ganglionectomy-induced changes were significant for SCG volume and mean perikaryal volume. These findings show that unilateral superior cervical ganglionectomy has profound effects on the contralateral ganglion. For future investigations, it would be interesting to examine the interaction between SCGs and their innervation targets after ganglionectomy. Is the ganglionectomy-induced imbalance between the sizes of innervation territories the milieu in which morphoquantitative changes, particularly changes in perikaryal volume and neuron number, occur? Mechanistically, how would those changes arise? Are there any grounds for believing in a ganglionectomy-triggered SCG cross-innervation and neuroplasticity?, (Copyright © 2011 ISDN. Published by Elsevier Ltd. All rights reserved.)

Ooplasm transfer has been used successfully to treat infertility in women with ooplasmic insufficiency and has culminated in the birth of healthy babies. To investigate whether mitochondrial dysfunction is a factor in ooplasmic insufficiency, bovine oocytes were exposed to ethidium bromide, an inhibitor of mitochondrial DNA replication and transcription, during in-vitro maturation (IVM). Exposure of immature oocytes to ethidium bromide for 24h during IVM hampered meiotic resumption and the migration of cortical granules. However, a briefer treatment with ethidium bromide during the last 4h of IVM led to partial arrest of preimplantation development without affecting oocyte maturation. Ooplasm transfer was then performed to rescue the oocytes with impaired development. In spite of this developmental hindrance, transfer of normal ooplasm into ethidium bromide-treated oocytes resulted in a complete rescue of embryonic development and the birth of heteroplasmic calves. Although this study unable to determine whether developmental rescue occurred exclusively through introduction of unaffected mitochondria into ethidium bromide-damaged oocytes, e.g. ethidium bromide may also affect other ooplasm components, these results clearly demonstrate that ooplasm transfer can completely rescue developmentally compromised oocytes, supporting the potential use of ooplasm transfer in therapeutic applications., (Copyright © 2010 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.)

This study was conducted to determine the relationship among temperatures measured at different anatomical sites of the animal body and their daily pattern as indicative of the thermal stress in lactating dairy cows under tropical conditions. Environmental dry bulb (DBT) and black globe (BGT) temperatures and relative humidity (RH) were recorded. Rectal temperature (RT), respiratory frequency (RF), body surface (BST), internal base of tail (TT), vulva (VT) and auricular temperatures (AT) were collected, from 37 Black and White Holstein cows at 0700, 1300 and 1800 hours. RT showed a moderately and positive correlations with all body temperatures, ranging from 0.59 with TT to 0.64 with BST. Correlations among AT, VT and TT with RF were very similar (from 0.63 to 0.64) and were greater than those observed for RF with RT (0.55) or with BST (0.54). RF and RT were positively correlated to TT (0.63 and 0.59, respectively), AT (r = 0.63 for both) and VT (r = 0.64 and 0.63, respectively). Positive and very high correlations were observed among AT, VT and TT (from 0.94 to 0.97) indicating good association of temperatures measured in these anatomical sites. Correlations of BST with AT and VT were positive and very similar (0.71 and 0.72, respectively) and lower with TT (0.66). The AT, TT, VT and BST presented similar patterns and follow the variations of DBT through the day. Temperatures measured at different anatomical sites of the animal body have the potential to be used as indicative of the thermal stress in lactating dairy cows.

The effect of genetic and non-genetic factors for carcass, breast meat and leg weights, and yields of a commercial broiler line were investigated using the restricted maximum likelihood method, considering four different animal models, including or excluding maternal genetic effect with covariance between direct and maternal genetic effects, and maternal permanent environmental effect. The likelihood ratio test was used to determine the most adequate model for each trait. For carcass, breast, and leg weight, and for carcass and breast yield, maternal genetic and permanent environmental effects as well as the covariance between direct and maternal genetic effects were significant. The estimates of direct and maternal heritability were 0.17 and 0.04 for carcass weight, 0.26 and 0.06 for breast weight, 0.22 and 0.02 for leg weight, 0.32 and 0.02 for carcass yield, and 0.52 and 0.04 for breast yield, respectively. For leg yield, maternal permanent environmental effect was important, in addition to direct genetic effects. For that trait, direct heritability and maternal permanent environmental variance as a proportion of the phenotypic variance were 0.43 and 0.02, respectively. The results indicate that ignoring maternal effects in the models, even though they were of small magnitude (0.02 to 0.06), tended to overestimate direct genetic variance and heritability for all traits.

Genetic parameters for traits related to postweaning growth in Braunvieh cattle, reared under tropical and sub-tropical conditions in Brazil, were studied. Weight traits were weight at 365 days of age (W365, N = 4055), at 450 days (W450, N = 3453), and at 550 days (W550, N = 1946), while weight gains were gain from weaning to 365 days of age (WGW365, N = 3060), from weaning to 450 days (WGW450, N = 2764), from weaning to 550 days (WGW550, N = 1531), from 365 to 550 days of age (WG365550, N = 1528), from 365 to 450 days (WG365450, N = 2401), and from 450 to 550 days (WG450550, N = 1563). A full animal model was used for estimating the variance components, using the MTDFREML software. The dataset contained 18,688 animals with phenotypic measures and 35,188 animals in the relationship matrix. Heritability estimates for postweaning weights decreased with age. For W365, W450 and W550, respectively, the direct heritability estimates were 0.29 +/- 0.061, 0.25 +/- 0.057, 0.16 +/- 0.060, maternal heritability was 0.20 +/- 0.035, 0.18 +/- 0.035, 0.13 +/- 0.052, and total heritability was 0.30, 0.35, 0.26. In this breed, maternal influence was found to be important up to 550 days of age. The greater genetic correlations between weights were observed for weights measured at shorter intervals. A large environmental effect was observed for weight gain between weaning and 550 days; this effect was greater for the gains between 365 and 550 days.

This study was aimed at assessing the changes in sperm motion patterns and the percentage of acrosome reaction (AR) in domestic cat semen after treatment with either ionomycin or progesterone (P(4)). Ten ejaculates were collected from five tomcats using an artificial vagina, and were diluted, centrifuged and resuspended in a capacitation medium. Samples were evaluated and divided into seven equal aliquots and, after 2 h at 25 degrees C, were incubated for 30 min at 38 degrees C in 5% CO(2) and then analyzed. Computer-assisted sperm analysis and a combination of three fluorescent probes were used to assess sperm plasma, acrosomal membrane integrity and mitochondrial transmembrane potential. Thirty minutes after the start of incubation, P(4) was added (10 microg/ml) to the P1 group. Groups P2 and P3 were supplemented with P(4) (10 and 20 microg/ml, respectively) only after 2 h of incubation, and groups I1 and I2 were supplemented with ionomycin (4 and 8 mum, respectively) 2 h after incubation. Group E was supplemented with ethanol (0.6%) at 2 h after incubation and group C received no supplementation. Ionomycin and P(4) treatments led to a hyperactivation-like sperm motion and an increase (p < 0.05) in the percentage of AR. Although a higher (p < 0.05) percentage of AR was obtained in group I2 when compared with all P(4) groups, a decrease (p < 0.05) in total and progressive motility was observed in I2 group. As I1 group was similar to I2 to induce AR without diminishing sperm motility, we can conclude that ionomycin at 4 microm seems to be more suitable to trigger AR in domestic cat sperm.

Data from the slaughter of 24,001 chickens that were part of a selection program for the production of commercial broilers were used to estimate genetic trend for absolute carcass (CW), breast meat (BRW), and leg (LW) weights, and relative carcass (CY), breast meat (BRY), and leg (LY) weights. The components of (co)variance and breeding values of individuals were obtained by the restricted maximum likelihood method applied to animal models. The relationship matrix was composed of 132,442 birds. The models included as random effects, maternal additive genetic and permanent environmental for CW, BRW, LW, CY, and BRY, and only maternal permanent environmental for LY, besides the direct additive genetic and residual effects, and as fixed effects, hatch week, parents' mating group and sex. The estimates of genetic trend were obtained by average regression of breeding value on generation, and the average genetic trend was estimated by regression coefficients. The genetic trends for CW (+6.0336 g/generation), BRW (+3.6723 g/generation), LW (+1.5846 g/generation), CY (+0.1195%/generation), and BRY (+0.1388%/generation) were positive, and they were in accordance with the objectives of the selection program for these traits. The genetic trend for LY (-0.0019%/generation) was negative, possibly due to the strong emphasis on selection for BRY and the negative correlations between these two traits.

The objective of this study was to estimate genetic parameters for pre-weaning traits of Braunvieh cattle raised under tropical conditions in Brazil. The weight and weight gain parameters were birth weight (BW, N = 9955), weight at 120 days of age (W120, N = 5901), weaning weight at 205 days (WW, N = 6970), weight gain from birth to 205 days (GAIN205, N = 6013), weight gain from birth to 120 days (GAIN120, N = 5135), and weight gain from 120 to 205 days (GAIN85, N = 4482). Variance components were estimated using the animal model with the MTDFREML software. The relationship matrix included 35,188 animals; phenotypic measures were available for 18,688. Direct and maternal heritability increased from birth to weaning, with estimates of 0.23 +/- 0.037, 0.25+/- 0.050, 0.41+/- 0.059 for direct heritability for BW, W120 and WW, respectively, 0.08 +/- 0.012, 0.15 +/- 0.032, 0.22 +/- 0.036 for maternal genetic effects, and 0.18, 0.14 and 0.16 for total heritability estimates. For pre-weaning gains, estimates of heritability were 0.36 +/- 0.059, 0.30+/- 0.059, 0.12 +/- 0.035 for direct genetic effects of the traits GAIN205, GAIN120 and GAIN85, respectively, 0.23 +/- 0.038, 0.17 +/- 0.037, 0.03 +/- 0.029 for estimates of maternal heritability, and 0.12, 0.13, 0.16 for total heritability, respectively. Genetic correlations between weights were greater between measures taken at shorter intervals. This information can be used to optimize the design of programs for genetic improvement of Braunvieh cattle raised under tropical conditions.

The aim of the present study was to evaluate the heterosis effects on weaning weight at 205 days (WW, n = 146,464), yearling weight at 390 days (YW, n = 69,315) and weight gain from weaning to yearling (WG, n = 59,307) in composite beef cattle. The fixed models were: RM, which included contemporary groups, class of age of dam, outcrossing percentages for direct and maternal effects, and additive direct and maternal (AM) breed effects; R, RM model, minus AM breed effects, and H, RM model, minus additive breed effects. The estimates for W205 were in general positive (P < 0.01). The R and H models resulted in similar estimates, but they were very different from the ones estimated by the RM model. For W390, the R and H models resulted in general positive estimates (P < 0.05). For WG, the RM model resulted in general significant heterosis effects (P < 0.05). It can be concluded that the RM model seems to supply estimates of better quality (P < 0.01).

Diabetes mellitus is the most common endocrine disturbance of domestic carnivores and can cause autonomic neurological disorders, although these are still poorly understood in veterinary medicine. There is little information available on the quantitative adaptation mechanisms of the sympathetic ganglia during diabetes mellitus in domestic mammals. By combining morphometric methods and NADPH-diaphorase staining (as a possible marker for nitric oxide producing neurons), type I diabetes mellitus-related morphoquantitative changes were investigated in the celiac ganglion neurons in dogs. Twelve left celiac ganglia from adult female German shepherd dogs were examined: six ganglia were from non-diabetic and six from diabetic subjects. Consistent hypertrophy of the ganglia was noted in diabetic animals with increase of 55% in length, 53% in width, and 61.5% in thickness. The ordinary microstructure of the ganglia was modified leading to an uneven distribution of the ganglionic units and a more evident distribution of axon fascicles. In contrast to non-diabetic dogs, there was a lack of NADPH-diaphorase perikarial labelling in the celiac ganglion neurons of diabetic animals. The morphometric study showed that both the neuronal and nuclear sizes were significantly larger in diabetic dogs (1.3 and 1.39 times, respectively). The profile density and area fraction of NADPH-diaphorase-reactive celiac ganglion neurons were significantly larger (1.35 and 1.48 times, respectively) in non-diabetic dogs compared to NADPH-diaphorase-non-reactive celiac ganglion neurons in diabetic dogs. Although this study suggests that diabetic neuropathy is associated with neuronal hypertrophy, controversy remains over the possibility of ongoing neuronal loss and the functional interrelationship between them. It is unclear whether neuronal hypertrophy could be a compensation mechanism for a putative neuronal loss during the diabetes mellitus.

Somatic nuclear transfer allows scientists to produce identical copies of individuals (clones) and offers the potential for research in various fields. However, the production of cloned bovines is inefficient, partly because of high rates of embryonic mortality associated with abnormal placental development, although the exact mechanisms are still unclear. In the present study, we have applied design-based stereological methods to quantify placental morphology in cloned and non-cloned Nelore bovines (280-297 days of gestation). We have found no differences for any of the following parameters: number and volume of placentomes, villous volume and surface area, volume and number of star volume units and placental efficiency. Therefore, placental deficiency is unlikely to be attributable to its villous component. Nevertheless, new studies should focus on other placental constituents, e.g. capillaries, trophoblast cells and feto-maternal exchange surface area.

The observation of bovine mitochondrial DNA (mtDNA) polymorphisms allows the separation of American zebu cattle, according to its maternal lineage ancestry, into two groups: one with Bos indicus mtDNA and other with Bos taurus mtDNA. The aim of the present study was to determine the productive and reproductive differences between these two groups, in a Guzerat dairy herd. The genotyping of a sample of 56 animals allowed the categorization of most of the 3835 animals in the pedigree file. The production file included 3528 calving and 3198 lactation records from 729 cows, born during the years 1947 to 2007. The traits considered were: lactation milk yield (LMY); days in milk (DIM); age at first calving (AFC), and calving interval (CI). Heritabilities and breeding values were estimated using an animal model. The regression of the average breeding values per year of birth indicated the genetic trends of the herd. The heritability coefficients estimated for LMY, DIM, AFC, and CI were 0.42, 0.43, 0.20, and 0.10, respectively. The genetic trends were similar for both groups, pointing to an improvement in the productive and a worsening in the reproductive traits. The two groups differed significantly regarding the average estimated breeding values for LMY, DIM and AFC, in the starting period, until 1970, but no differences were observed in the more recent years, after 1970. The segregation between the groups existed in the starting period, probably because the Bos taurus contributions to the herd had occurred more recently at that moment. The conclusion is that mtDNA has no significant effect on these traits.

The purpose of the present research was to investigate the effects of polymorphisms of luteinizing hormone receptor (LHR) and follicle-stimulating hormone receptor (FSHR) genes, evaluated by polymerase chain reaction-restriction fragment length polymorphism in European-Zebu composite beef heifers from six different breed compositions. The polymorphism site analysis from digestion with HhaI and AluI restriction endonucleases allowed the genotype identification for LHR (TT, CT and CC) and FSHR (GG, CG and CC) genes. A high frequency of heterozygous animals was recorded in all breed compositions for both genes, except in two compositions for LHR. The probability of pregnancy (PP) at first breeding was used to evaluate the polymorphism effect on sexual precocity. The PP was analyzed as a binary trait, with a value of 1 (success) assigned to heifers that were diagnosed pregnant by rectal palpation and a value of 0 (failure) assigned to those that were not pregnant at that time. Heterozygous heifers showed a higher pregnancy rate (67 and 66% for LHR and FSHR genes, respectively), but no significant effects were observed for the genes studied (P = 0.9188 and 0.8831 for LHR and FSHR, respectively) on the PP. These results do not justify the inclusion of LHR and FSHR restriction fragment length polymorphism markers in selection programs for sexual precocity in beef heifers. Nevertheless, these markers make possible the genotype characterization and may be used in additional studies to evaluate the genetic structure in other bovine populations.