Maternal rumen-protected methionine supplementation and its effect on blood and liver biomarkers of energy metabolism, inflammation, and oxidative stress in neonatal Holstein calves

In nonruminants, nutrition during pregnancy can program offspring development, metabolism, and health in later life. Rumen-protected Met (RPM) supplementation during the prepartum period improves liver function and immune response in dairy cows. Our aim was to investigate the effects of RPM during late pregnancy on blood biomarkers (23 targets) and the liver transcriptome (24 genes) in neonatal calves from cows fed RPM at 0.08% of diet dry matter/d (MET) for the last 21 d before calving or controls (CON). Blood (n=12 calves per diet) was collected at birth before receiving colostrum (baseline), 24 h after receiving colostrum, 14, 28, and 50 d (post-weaning) of age. Liver was sampled (n=8 calves per diet) via biopsy on d 4, 14, 28, and 50 of age. Growth and health were not affected by maternal diet. The MET calves had greater overall plasma insulin concentration and lower glucose and ratios of glucose-to-insulin and fatty acids-to-insulin, indicating greater systemic insulin sensitivity. Lower concentration of reactive oxygen metabolites at 14 d of age along with a tendency for lower overall concentration of ceruloplasmin in MET calves indicated a lesser degree of stress. Greater expression on d 4 of fructose-bisphosphatase 1 (FBP1), phosphoenolpyruvate carboxykinase 1 (PCK1), and the facilitated bidirectional glucose transporter SLC2A2 in MET calves indicated alterations in gluconeogenesis and glucose uptake and release. The data agree with the greater expression of the glucocorticoid receptor (GR). Greater expression on d 4 of the insulin receptor (INSR) and insulin-responsive serine/threonine-protein kinase (AKT2) in MET calves indicated alterations in insulin signaling. In that context, the similar expression of sterol regulatory element-binding transcription factor 1 (SREBF1) in CON and MET during the preweaning period followed by the marked upregulation regardless of diet after weaning (d 50) support the idea of changes in hepatic insulin sensitivity during early postnatal life. Expression of carnitine palmitoyltransferase 1A (CPT1A) was overall greater and acyl-CoA oxidase 1 (ACOX1) was lower in MET calves, indicating alterations in fatty acid oxidation. Except forkhead box O1 (FOXO1), all genes changed in expression over time. Transcriptome results indicated that calves from MET-supplemented cows underwent a faster maturation of gluconeogenesis and fatty acid oxidation in the liver, which would be advantageous for adapting to the metabolic demands of extrauterine life

VARIACIONES DIARIAS DE TEMPERATURA Y SU IMPACTO EN LA PRODUCCIÓN DE LECHE EN ALTURA

Extensive dairy productions are exposed to weather variations, and when the temperature and humidity are extreme, animals can experience thermal stress. The aim of this work was to evaluate the impact of a daily variation on temperature-humidity index (THI) on circulating cortisol levels, milk yield and composition of dairy cows. Ten Holstein cows in the first third of lactation were evaluated, during morning and afternoon milking, for milk yield, milk composition and blood cortisol. Climatic variables were recorded daily. During the experimental period, the daily temperature variations were around 15?C (range between maximum and minimum), however THI indicated animals were in a thermoneutral zone. Milk composition and blood cortisol concentrations were not different between morning and afternoon milking and blood cortisol concentration had a weak positive correlation with milk yield and negative with milk protein, fat and non-fat solids content.  In conclusion, Holstein cows located in regions of high altitude do not show a significant level of relationship between blood cortisol and production and milk components, which may indicate the adaptation of these animals to this environmental condition.Los sistemas ganaderos extensivos están expuestos a las variaciones climáticas, y cuando la temperatura y humedad son extremas, los animales pueden enfrentar estrés térmico. El objetivo de este estudio fue evaluar el impacto de la variación diaria de la temperatura y el índice temperatura y humedad (ITH) sobre las concentraciones sanguíneas de cortisol, producción y composición de leche en vacas Holstein. Diez vacas fueron seleccionadas, evaluadas en el primer tercio de lactancia, durante los ordeños de la mañana y tarde, para analizar concentraciones sanguíneas de cortisol, producción y composición de leche. Las variables climáticas fueron reportadas diariamente. Durante el periodo experimental, la variación diaria de temperatura estuvo al redor de 15?C (rango entre máxima y mínima), pero el ITH indicó que los animales estaban en una zona termoneutral. La composición de la leche y la concentración sanguínea de cortisol no fue diferente entre los ordeños de la mañana y tarde y el cortisol sanguíneo tuvo una correlación baja, positiva con producción de leche y negativa con la concentración de proteína, grasa y sólidos de la leche. En conclusión, vacas Holstein ubicadas en regiones de alta altitud no evidencian un nivel significativo de relación entre cortisol sanguíneo con la producción y componentes de la leche, lo que puede indicar adaptación de estos animales a esta condición ambiental

Metabolic and Ruminal Fluid Markers of Dairy Cows Supplemented with a Combination of Yeast Culture and Hydrolyzed Yeast

Background: In order to reduce the effects of a negative energy balance, some measures have been taken into account in nutritional management during the transition period. The use of yeast, has been a good alternative used to improve the rumen metabolism and helping the adjustment of the microbiotato the new diet. The aim of the study was to evaluate the effects of supplementing a combination of yeast culture and hydrolyzed yeast on the metabolism of dairy cows during the transition period.Materials, Methods & Results: The experiment was conducted in a semi-extensive system, using 20 Holstein cows, divided equally into a control group (CG) and a supplemented group (SG). The SG received 28 g/animal/day of a combination of yeast culture and hydrolyzed yeast from 20 ± 2 days pre-calving until early lactation (18 ± 3 days). Serum concentrations of non-esterified fatty acids (NEFA), albumin and urea were determined at calving, and for three time points during the early postpartum period and three time points during the early lactation period. Regarding energy metabolism, prepartum concentrations of NEFA were higher than the physiological standard in both groups. However, NEFA, albumin and urea decreased during the early postpartum period in the supplemented animals and could be attributed to the yeast in enhancing ruminal microorganisms’ cellulolytic capacity, increasing fibre digestibility and starch utilization.Discussion: The increased concentration of non-esterified fatty acids (NEFA) due to the mobilization of fat deposits that happens in the transition period, especially in the postpartum period reflects the cow’s adaptation to the negative energy balance (NEB). The lower concentrations of NEFA observed in the present study could be attributed to the effect of the yeast in enhancing the ruminal microorganisms’ cellulolytic capacity. The control cows had a BCS within the recommended range while the supplemented group had it close to the minimal limit proposed for this period. Thus, supplemented cows lost less BCS during the early postpartum period, had a lower BCS loss during the experimental period and had lower NEFA concentration that the CG. It was possible to observe a difference in serum albumin and urea between treatments only in the postpartum period. Besides showing no significant effect in BCS on prepartum period, control cows had a BCS within the recommended range while the supplemented group had it close to the minimal limit proposed for this period. Cows with high BCS prepartum had higher plasma NEFA before and after calving. It can be observed in the present study in both groups. However, a positive effect in prevent subclinical disorders might be attributed to YC, since the SG showed low NEFA plasma levels compared to the CG.  Thus, supplemented cows lost less BCS during the early postpartum period, had a lower BCS loss during the experimental period and had lower NEFA concentration that the CG. There is a negative correlation between BCS and NEFA in the early postpartum period and this information explains the results observed in the present study where BCS declines in the SG are followed by a NEFA increase. This is not so marked in the CG, indicating that SG supplementation can act by improving digestibility. Yeast supplementation promotes higher output energy, enhancing postpartum performance in dairy cows. Yeast supplementation showed benefits in early lactation compared to the prepartum and early postpartum periods, suggesting that supplementation has to have an adaptation period to be effective in protein synthesis. In conclusion, supplementation with a combination of yeast culture and hydrolyzed yeast to cows during the transition period can positively influence the energy and protein metabolism, reducing the collateral effects of negative energy balance

Variabilidade genética em tambaquis (Teleostei: Characidae) de diferentes regiões do Brasil

The objective of this study was to evaluate the genetic diversity of four broodstocks of tambaqui (Colossoma macropomum) from different regions of Brazil using RAPD markers. Ten primers were used to analyze 116 individuals, collected from fish cultures of three municipalities in Brazil: Urupá, RO; Teixeirópolis, RO; Neópolis, SE; and Sorriso, MT. Differences in the frequencies of 67 fragments were found, with a unique fragment in Sorriso and two in Neópolis. High values of polymorphism (72.92 to 83.33%), Nei's genetic diversity (from 0.27 to 0.30) and Shannon index (from 0.39 to 0.45) were observed. Analysis of molecular variance showed that most of the variation is within each broodstock and not among them. The identity and genetic distance among the groups ranged from 0.93 to 0.98 and from 0.02 to 0.07, respectively, with less distance between clusters Urupá x Sorriso and Teixerópolis x Neópolis. Genetic differentiation ranged from low to moderate (Fst= 0.03 to 0.15) and the number of migrants per generation was high (Nm = 5.96 to 24.3) among the groups. Stocks have high variability and low genetic differentiation and distance among them.O objetivo deste trabalho foi avaliar a diversidade genética em quatro estoques de tambaquis (Colossoma macropomum) de diferentes regiões do Brasil, por meio de marcador RAPD. Foram utilizados 10 iniciadores para analisar 116 indivíduos, coletados de pisciculturas nos municípios de Urupá, RO, Teixeirópolis, RO, Neópolis, SE e Sorriso, MT. Foram encontradas diferenças nas frequências de 67 fragmentos, com um fragmento exclusivo em Sorriso e dois em Neópolis. Observaram-se altos valores de polimorfismo (72,92 a 83,33%), diversidade genética de Nei (0,27 a 0,30) e índice de Shannon (0,39 a 0,45). A análise da variância molecular, demonstrou que a maior parte da variação está dentro de cada estoque e não entre os estoques. A identidade e a distância genética entre os agrupamentos variou de 0,93 a 0,98 e 0,02 a 0,07, respectivamente, com menos distância entre os agrupamentos Urupá x Sorriso e entre Teixerópolis x Neópolis. A diferenciação genética variou de baixa a moderada (Fst = 0,03 a 0,15) e o número de migrantes por geração foi alto (Nm = 5,96 a 24,3), entre os agrupamentos. Os estoques apresentam alta variabilidade e baixa diferenciação e distância genética entre si

Nutrimetabolomics: effects of maternal supplementation with organic minerals or methionine on metabolism and immunometabolic programming of neonatal calves

Understanding how markers of hepatic and immune metabolism are regulated during the neonatal period and how they could be affected by pre-partum maternal nutrition contributes to the development of nutritional strategies that could ameliorate the initial stress of birth. Transcriptomics is a tool that has been recently applied aiming to better understand the molecular mechanisms during stressful periods. Although the neonatal calf physiology has been well-studied, the mechanisms which pre-partum nutrition can affect the hepatic and immune metabolism of the neonatal calf still warrant further investigations. The aim of this thesis was to evaluate the effect of pre-partum maternal supplementation with organic trace minerals or methionine on neonatal calf metabolism and immune function. Maternal supplementation with organic trace minerals during the last 30 days of pregnancy enhanced neonatal immune function, reducing the pro-inflammatory state and regulating the oxidative stress, during the first 3 weeks of life. However, maternal supplementation with methionine during the last 24 days of pregnancy did not affect calves immune metabolism, from birth through 50 days of age. Regarding hepatic metabolism, maternal supplementation with methionine during the last 24 days of pregnancy improved calves hepatic metabolism, having greater maturation of the metabolic pathways and also higher efficiency in glucose uptake and endogenous production, from birth through 50 days of age. According to the currently available literature, these are the first studies demonstrating how maternal supplementation with organic trace minerals or methionine acts regulating the immune system and hepatic metabolism of neonatal calves, focusing on gene expression and blood biomarkers. In conclusion, the results indicate that materal prepartum supplementation with organic trace minerals or methionine elicts some beneficial adaptations in the immune and hepatic metabolism of neonatal calves, contributing to a better adaptation to the extra-uterine environment.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESO conhecimento de como marcadores do metabolismo imune e hepático são regulados durante o período neonatal e como podem ser influenciados pela dieta materna pré-parto contribui para o desenvolvimento de estratégias nutricionais, buscando reduzir o estresse do nascimento. A transcriptomica é uma ferramenta que tem sido utilizada recentemente com intuito de contribuir para o melhor entendimento dos mecanismos moleculares que regulam o metabolismo durante períodos de estresse. Embora a fisiologia neonatal bovina já seja bem compreendida, os mecanismos pelos quais a nutrição materna no pré-parto pode influenciar o metabolismo hepático e imune dos neonatos bovinos ainda necessitam de maiores investigações. O objetivo desta tese foi avaliar o efeito da suplementação materna no pré-parto com microminerais orgânicos ou metionina no metabolismo e capacidade imune de neonatos bovinos. A suplementação materna com microminerais orgânicos durante os últimos 30 dias de gestação melhorou a função imune dos neonatos, reduzindo o estado pró-inflamatório e controlando o estresse oxidativo, durante as primeiras três semanas de vida. Entretado, a suplementação materna com metionina durante os últimos 24 dias pré-parto não teve efeito sobre o metabolismo imune dos bezerros, do nascimento aos 50 dias de idade. Em relação ao metabolismo hepático, a suplementação materna com metionina durante os últimos 24 dias pré-parto apresentou um efeito benéfico sobre o metabolismo hepático dos bezerros, proporcionando uma maturação mais evidente das rotas metabólicas e também maior eficiência na captação e produção endógena de glicose, do nascimento aos 50 dias de idade. De acordo com a bibliografia atualmente disponível, estes são os primeiros trabalhos a demonstrarem como a suplementação materna com microminerais orgânicos ou metionina atua na regulação do sistema imune e metabolismo hepático de neonatos bovinos, focando em expressão gênica e biomarcadores sanguíneos. Em conclusão, os resultados obtidos evidenciam que a suplementação materna pré-parto com microminerais orgânicos ou metionina pode beneficiar o metabolismo imune e hepático dos neonatos bovinos, contribuindo para uma melhor adaptação ao ambiente extra-uterino

Nutrigenomics: Effect of linolenic acid containing diets on gene expression of nuclear receptors related to lipid metabolism

Dietary polyunsaturated fatty acids (PUFAs) are negative regulators of hepatic lipogenesis and their effects are at transcriptional levels. The essential fatty acids omega-3 and omega-6, consumed during pregnancy can benefit maternal and offspring health. The nuclear receptor peroxisome proliferator-activated receptor α (PPARα), when activated by PUFAs enhances lipolytic genes. Liver X receptor α (LXRα) and sterol regulatory element binding protein 1c (SREBP-1c) are nuclear receptors that enhance lipogenic genes, and are repressed by PUFAs. The present study aimed to investigate the effects of diets, rich in omega-3 or omega-6, consumed throughout three generations, on biochemical parameters and the expression level of some genes related to lipid metabolism. For this study we used adult Wistar/UFPel rats, composing the founding generation, females that received diet with flaxseed oil (OM group, n=18) or soybean oil (CTL group, n=18) during pregnancy. At F1 weaning, 48 females offspring were selected and allocated in three groups, females from OM group, that continued receiving diet with flaxseed oil (OM/OM group, n=16), females from OM group, that started to receive diet with soybean oil (OM/CTL group, n=16) and females from CTL group, that continued receiving diet with soybean oil (CTL/CTL group, n=16). At F2 weaning, 16 females offspring of each group were selected and continued receiving the same diets (OM/OM/OM group, OM/CTL/CTL group and CTL/CTL/CTL group). The females were evaluated in the pre-partum (19±1 days of pregnancy) and at postpartum period (21 days after parturition). The dietary fatty acid profile did not affect the diet daily intake and the pregnancy rate (P>0.05), however, dams that received the diet rich in omega-3 had heaviest offspring (P=0.01). Diets with higher omega-3 level decreased triglycerides serum levels, and maintained constant the non-esterified fatty acids (NEFA) levels (P=0.04), while the animals fed with high proportion of omega-6 had fluctuant NEFA levels, and an outstanding increase in the postpartum period of F2 generation (P=0.02). PPARα, RXRα, LXRα and SREBP-1c had been regulated by the diets throughout generations. In the pre-partum moment of F2 generation, the animals fed with high omega-3 had a lower expression of LXRα (P=0.01) indicating its effectiveness on inhibiting lipogenic action. Our results indicate that the PUFAs effect on the control of lipolysis and lipogenesis is cumulative throughout generations and the omega-3 exerts a better control.Dietas contendo ácidos graxos poli-insaturados (AGPIs) exercem controle negativo sobre a lipogênese hepática e seus efeitos são observados ao nível transcricional. Os ácidos graxos essenciais ômega-3 e ômega-6, quando consumidos durante a gestação podem beneficiar a saúde materna e da prole. O receptor nuclear peroxisome proliferator-activated receptor α (PPARα), quando ativado pelos AGPIs favorecem a atividade de genes lipolíticos. O liver X receptor α (LXRα) e sterol regulatory element binding protein 1c (SREBP-1c) são receptors nucleares que ativam genes lipogênicos, e são reprimidos pelos AGPIs. O presente estudo teve como objetivo investigar os efeitos de dietas, ricas em ômega-3 ou ômega-6, consumidas ao longo de três gerações, sobre parâmetros bioquímicos e o nível de expressão de alguns genes relacionados ao metabolismo lipídico. Para este estudo foram utilizados ratas adultas, da linhagem Wistar/UFPel, compondo a geração fundadora, fêmeas que receberam ração com óleo de linhaça (Grupo OM, n=18) ou óleo de soja (Grupo CTL, n=18) durante toda a gestação. No desmame da F1 foram selecionadas 48 progênies fêmeas, então divididas em três grupos, fêmeas oriundas do Grupo OM, que continuaram a receber dieta com óleo de linhaça (Grupo OM/OM, n=16), fêmeas oriundas do Grupo OM, que passaram a receber dieta com óleo de soja (Grupo OM/CTL, n=16) e fêmeas oriundas do Grupo CTL, que continuaram a receber dieta com óleo de soja (Grupo CTL/CTL, n=16). No desmame da F2 foram selecionadas 16 progênies fêmeas de cada grupo que seguiram recebendo o mesmo tratamento (Grupo OM/OM/OM; Grupo OM/CTL/CTL e Grupo CTL/CTL/CTL). As fêmeas foram avaliadas no período pré parto (19±1 dias de gestação) e no pós-parto (21 dias após o parto). O perfil de ácidos graxo da dieta não afetou o consumo de ração e nem a taxa de prenhez (P>0,05), porém as fêmeas alimentadas com dieta rica em ômega-3 tiveram filhotes mais pesados (P=0,01). A dieta com alto nível de ômega-3 diminuiu os níveis séricos de triglicerídeos (P=0,04), e mantiveram constante os níveis de ácidos graxos não-esterificados (AGNEs), enquanto animais alimentados com uma proporção maior de ômega-6 tiveram níveis flutuantes de AGNEs, e um notável aumento no perído pós-parto da geração F2 (P=0,02). PPARα, RXRα, LXRα and SREBP-1c foram regulados pelas dietas ao longo das gerações. No período pré-parto da geração F2, os animais alimentação com alto ômega-3 apresentaram uma menor expressão de LXRα (P<0,001), indicando sua efetividade em inibir a ação lipogênica. Nossos resultados indicam que os efeitos dos AGPIs no controle da lipólise e lipogênese é cumulativo ao longo das gerações e o ômega-3 exerce um controle mais efetivo