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1.
J Dairy Sci ; 106(12): 9532-9551, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37678778

RESUMEN

Study aims were to investigate associations of hyperketolactia (HYKL) status of Holstein dairy cows between 6 and 60 d in milk (DIM), defined by milk acetone (mACE) and ß-hydroxybutyrate (mBHB) content, with daily milk yield and composition. Milk samples (∼5.0 million) were collected over a 5-yr period (2014-2019) within the milk recording system in Poland. Concentrations of mACE and mBHB determined by Fourier-transform infrared spectroscopy were used to categorize samples into 4 ketolactia groups. Based on threshold values of ≥0.15 mmol/L mACE and ≥0.10 mmol/L mBHB, ketolactia groups were normoketolactia (NKL; mACE <0.15 mmol/L and mBHB <0.10 mmol/L), BHB hyperketolactia (HYKLBHB; mACE <0.15 mmol/L and mBHB ≥0.10 mmol/L), ACE hyperketolactia (HYKLACE; mACE ≥0.15 mmol/L and mBHB <0.10 mmol/L), and ACE and BHB hyperketolactia (HYKLACEBHB; mACE ≥0.15 mmol/L and mBHB ≥0.10 mmol/L). To investigate ketolactia association with production outcomes, a linear model was developed, including ketolactia group, DIM, parity, their interactions, year-season as fixed effects, and random effects of herd and cow. Among all milk samples, 31.2% were classified as HYKL, and of these, 52.6%, 39.6%, and 7.8% were HYKLACEBHB, HYKLBHB, and HYKLACE, respectively. Ketolactia groups differed for all traits studied in all parities and DIM. Among HYKL groups, lowest milk yield was found in HYKLACEBHB cows, except for 6 to 30 DIM in first- and second-lactation cows. Milk yield of HYKLBHB cows was higher than that of NKL cows until 20 to 30 DIM, and then it was lower than NKL cows. Milk yield of HYKLACE cows was mostly lower than NKL cows. Energy-corrected milk (ECM) yield of HYKLACEBHB cows was higher than that of NKL cows until 30 to 35 DIM for second lactation and third lactation or greater, and in the whole study period for first lactation. The yield of ECM for HYKLBHB cows was mostly higher than that of NKL cows, whereas HYKLACE cows had higher ECM than NKL cows until 15 to 25 DIM and then was lower for the HYKLACE group. Milk composition differed among HYKL groups. Highest milk fat (MF) and lowest milk lactose (ML) contents were observed in HYKLACEBHB cows. Cows in HYKLACEBHB and HYKLBHB groups had higher MF and lower milk protein (MP; except in 6-8 DIM in first lactation) and ML content than NKL cows. Milk fat content was higher in HYKLACE than NKL cows in first lactation and during the first 30 to 40 DIM in older cows. Lactose content was lower in HYKLACE than in NKL cows within 30 to 40 DIM; afterward it was higher in NKL cows. Lower MP content was found in HYKLACE than in NKL cows, except during 6 to 9 DIM for cows in first lactation and third lactation or greater. In conclusion, HYKL is associated with altered milk production in all parities, but a range of these negative relations depends on ketone status addressing both ACE and BHB contents. Further research is needed to ascertain underpinning biochemical defects of HYKL from elevated ACE, alone or in combination with BHB, during early lactation.


Asunto(s)
Lactancia , Lactosa , Embarazo , Femenino , Bovinos , Animales , Lactosa/análisis , Leche/química , Cetonas/análisis , Acetona/análisis , Ácido 3-Hidroxibutírico/metabolismo
2.
J Dairy Sci ; 104(12): 12800-12815, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-34538496

RESUMEN

Fourier transform infrared spectroscopy (FTIR) allows for the determination of milk acetone (mACE) and ß-hydroxybutyrate (mBHB) concentrations, providing a potential herd monitoring tool for hyperketolactia, defined as elevated milk ketone bodies. The study aim was to characterize mACE and mBHB concentration dynamics during early lactation in Polish Holstein-Friesian cows. Milk samples (n = 3,867,390) were collected within 6 to 60 days in milk (DIM) over a 4-yr period (April 1, 2013 to March 31, 2017) from approximately 21,300 dairy herds (average 38.7 cows/herd). Fixed effects of parity, DIM, and their interaction on mACE and mBHB concentrations were determined using a mixed model with a herd-year-season fixed effect and random cow effect. Published hyperketolactic mACE (≥0.15 mmol/L) and mBHB (≥0.10 mmol/L) threshold concentrations were used to classify study milk samples into ketolactia groups of normal (mACE <0.15 mmol/L and mBHB <0.10 mmol/L) and hyperketolactic (HYKL; either mACE ≥0.15 mmol/L or mBHB ≥0.10 mmol/L). Additionally, HYKL samples were categorized into subpopulations as having elevated mBHB and mACE (HYKLACEBHB, mACE ≥0.15 mmol/L and mBHB ≥0.10 mmol/L), only elevated mBHB (HYKLBHB; mACE <0.15 mmol/L and mBHB ≥0.10 mmol/L), or only elevated mACE (HYKLACE; mACE ≥0.15 mmol/L and mBHB <0.10 mmol/L). Effects of parity, DIM, ketolactia group or subpopulation, and their interactions on mACE and mBHB concentrations were also determined using the mixed model that included ketolactia group or subpopulation as an independent variable. Across all data, mACE and mBHB concentrations were influenced by effects of parity, DIM, and their interaction as well as parity, DIM, ketolactia group or subpopulation, and their interactions. For all samples, mACE and mBHB concentrations decreased with increasing DIM, with mACE concentration declining more rapidly compared with mBHB. In the data set, 68% and 32% of all samples were defined as normal or HYKL, respectively. Among HYKL samples, mACE was elevated soon after calving and declined over time. In contrast, mBHB started lower after calving and increased reaching peak concentrations around 30 DIM, and then decreased. Within HYKL samples, 50.8, 41.3, and 7.9% were categorized as HYKLACEBHB, HYKLBHB, and HYKLACE respectively. Between 6 and 21 DIM, 11.3% of HYKL were classified as HYKLACE. Primiparous cows had greater (14.8%) HYKLACE samples in this time period. In conclusion, this study has characterized mACE and mBHB concentrations during early lactation and determined effects of parity, DIM, and their interaction. Using published criteria interpreting mACE and mBHB concentrations, it was intriguing to identify a unique population of samples having elevated mACE without mBHB in early lactation, especially in primiparous cows. Further research is needed to determine if this sample population represents an unhealthy metabolic status that adversely affects cow health and performance.


Asunto(s)
Enfermedades de los Bovinos , Ácido 3-Hidroxibutírico , Animales , Bovinos , Femenino , Lactancia , Leche , Paridad , Embarazo
3.
J Dairy Sci ; 104(7): 7725-7737, 2021 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-33838892

RESUMEN

Nitrogen balance (NB) experiments allow calculation of N retention in the body by subtracting N excreted in feces (NF), urine (NU) and milk (NM) from N intake (NI). In a previous study, we found that NB data from experiments with lactating dairy cows were generally high and, in the current meta-analysis, we update our earlier study with experiments from the last 2 decades and investigate probable causes of error. A total of 83 publications, with 86 experiments and 307 dietary treatments, were selected from top-ranked scientific journals that reported all NB components. The NB and NB components were analyzed by linear regression with a model that used NI as an independent variable and experiment as a random effect. The NF, NU and NM each represented 27 to 34% of NI, and the remaining N accumulated in the body was equal to 38.5 g/d (overall SD = 43.2 g/d). Retained N (as g/d or % of NI) increased linearly with NI, and this led to unlikely high N retentions, especially at high NI. Both NF and NU (g/d) increased with increasing NI, and we assume that some N in feces and urine were unaccounted. Only ~22% of experiments measured N in wet feces samples and, when analysis used dry samples, no mention of corrections due to potential volatile N losses during drying were reported. No experimentalists preserved feces immediately to prevent volatilization during collection. Moreover, ~27% of experiments estimated urine volumes by concentration of creatinine in spot samples, and in these experiments, NU was ~12% lower than those where total urine was collected (168 vs. 191 g/d). Only 40 experiments reported the volume and concentration of acids used for urine preservation, 33 furnished incomplete information, and the remainder did not describe the urine preservation method. In conclusion, the results of NB experiments using lactating dairy cows overestimate N retention, and the losses of N from feces and urine are the most probable reason.


Asunto(s)
Lactancia , Nitrógeno , Animales , Bovinos , Dieta/veterinaria , Heces/química , Femenino , Leche/química , Nitrógeno/análisis , Rumen/química
4.
J Dairy Sci ; 104(6): 6663-6676, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33685670

RESUMEN

The aim of this study was to assess the effect of protein source, either soybean meal (SM) or canola meal (CM), and microencapsulated sodium butyrate (MSB) supplementation in a pelleted starter mixture on the development of the gastrointestinal tract (GIT) in dairy calves. Twenty-eight bull calves (8.7 ± 0.8 d of age and 43.0 ± 4.4 kg; mean ± SD) were assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement: CM as a main source of protein without or with MSB or SM without or with MSB. Calves were fed starters ad libitum and exposed to a gradual weaning program, with weaning taking place on 51.7 ± 0.8 d of age. Calves were observed for an additional 3 wk after weaning and slaughtered on d 72.1 ± 0.9 of age, after which the GIT was dissected. Morphometric measurements were recorded, and samples for determination of ruminal fermentation, histology, gene expression, and brush border enzyme activities were collected. Canola meal use in the starter mixture increased abomasal tissue weight, jejunal tissue weight and length, and mRNA expression of SLC16A4 (formerly known as MCT4) and FFAR2 (GPR43) in the ruminal epithelium, and decreased ruminal ammonia and mRNA expression of SLC15A2 (PEPT2) and SLC6A14 (ATB0+) in the proximal small intestine and ileum, respectively. However, MSB inclusion in the starter mixture decreased ruminal papillae length, ruminal epithelial surface, and ruminal epithelium dry weight, while increasing mRNA expression of SLC16A1 (MCT1) in ruminal epithelia. Reduced ruminal surface area associated with MSB supplementation was the most apparent when MSB was combined with CM in the starter mixture. Additionally, MSB supplementation decreased the thickness of omasal epithelium, omasal epithelium living strata, and stratum corneum, and increased duodenal and ileal aminopeptidase A enzymatic activity and ileal aminopeptidase N enzymatic activity. Overall, CM might increase growth of the GIT of calves, particularly of the small intestine, but may negatively affect intestinal epithelium function and peptide and AA absorption. Supplementation of MSB has a negative effect on the ruminal and omasal epithelium development, particularly when combined in a starter mixture with CM.


Asunto(s)
Alimentación Animal , Rumen , Alimentación Animal/análisis , Animales , Ácido Butírico , Bovinos , Dieta/veterinaria , Suplementos Dietéticos , Tracto Gastrointestinal , Masculino , Glycine max , Destete
5.
J Dairy Sci ; 104(6): 6646-6662, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33685691

RESUMEN

Two studies were conducted to assess the effect of protein source and microencapsulated sodium butyrate (MSB) inclusion in pelleted starter mixtures on growth performance, gain to feed (G:F) ratio, nutrient digestibility, and selected blood metabolites in calves. In study 1, 28 Holstein bull calves (8.7 ± 0.8 d of age and 43.0 ± 4.4 kg; mean ± SD) were allocated to 1 of 4 treatments in a 2 × 2 factorial arrangement and fed a pelleted starter mixture containing canola meal (CM, 35% as fed) or soybean meal (SM, 24% as fed) as the main source of protein, with or without supplemental MSB (0.3% as fed). Starter mixtures were formulated to be similar for crude protein, Lys, and Met, and were fed ad libitum. Calves were weaned after 42 d of milk replacer feeding (51.7 ± 0.8 d of age) and observed for another 21 d. Furthermore, selected blood metabolites were measured on d 21, 42, and 63 of the study, and nutrient digestibility was measured after weaning. In study 2, 60 Holstein heifer calves (9.1 ± 0.8 d of age and 43.2 ± 4.2 kg) were assigned to the same treatments as in study 1. The calves were weaned after 49 d of milk replacer feeding (59.1 ± 0.8 d of age) and observed for an additional 14 d. Milk replacer and starter mixture intake and fecal score were recorded daily, whereas body weight (BW) was recorded weekly. In study 1, calves fed starter mixtures containing CM had or tended to have lesser preweaning starter intake, weaning average daily gain (ADG), weaning and overall G:F ratio, and postweaning total-tract dry matter digestibility, as opposed to those fed starter mixtures with SM. However, these differences did not affect overall starter intake, overall ADG, or final BW. Supplementation with MSB only tended to increase the preweaning starter mixture intake. In study 2, heifer calves that were fed starter mixtures with CM had greater cumulative starter intake after weaning, but the protein source in the starter mixture had no effect on ADG, BW, or G:F ratio. Inclusion of MSB in starter mixtures for calves tended to decrease postweaning starter mixture intake. In conclusion, use of CM or SM as the main source of protein in starter mixture resulted in similar growth performance of bull and heifer calves; however, CM use in starter mixtures reduced starter intake, ADG, and G:F ratio at least at some points of rearing. Supplementation of MSB had minor effects on the growth performance of calves.


Asunto(s)
Alimentación Animal , Glycine max , Alimentación Animal/análisis , Animales , Peso Corporal , Ácido Butírico , Bovinos , Dieta/veterinaria , Suplementos Dietéticos , Femenino , Masculino , Destete
6.
J Dairy Sci ; 103(9): 7998-8019, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32622603

RESUMEN

The objectives of this study were to assess the effect of using heat-treated canola meal (CM) and glycerol inclusion in starter mixtures on starter intake, growth, and gastrointestinal tract development in Holstein bull calves. In the first study, a protocol for the heat treatment of CM was evaluated by comparing commercial CM that was exposed to 0, 100, 110, or 120°C of heat treatment for 10 min. Following heat treatment, in situ crude protein (CP) ruminal degradability and estimated intestinal CP digestibility were assessed. It was observed that the degradable fractions of dry matter and CP in CM decreased linearly with increasing temperature of heat treatment. The estimated intestinal CP digestibility was greatest when CM was heated to 110°C. In the second study, 28 bull calves were used in a randomized complete block design. Calves were fed pelleted starters containing CM or CM that was heat-treated to 110°C for 10 min. Diets also contained 0 or 5% glycerol on a dry matter basis. The study lasted 51 d, ending on the first day of weaning. Starter intake, average daily gain (ADG), ruminal short-chain fatty acid concentrations, morphology of the rumen and small intestine, gene expression (MCT1, GPR41, GPR43, UTB, AQP3, PEPT1, PEPT2, ATB0+, and EAAC1) in the ruminal, jejunal, and ileal epithelium, and brush border enzyme activities in the duodenum, jejunum, and ileum were investigated. Few interactions between heat-treated CM and glycerol inclusion were observed. Feeding heat-treated CM did not affect starter intake. However, feeding heat-treated CM to calves tended to reduce ADG and decreased the weight of ruminal and jejunal tissue. Heat treatment did not affect gene expression or brush border enzyme activities in the small intestine. Glycerol inclusion tended to increase cumulative starter intake and increased cumulative body weight gain. Use of glycerol reduced ruminal pH and increased the concentration of ruminal short-chain fatty acids. Additionally, glycerol inclusion increased abomasal, duodenal, jejunal, and cecal digesta weights and tended to increase the weight of the jejunal tissue. Glycerol supplementation tended to downregulate the expression of MCT1 in the ruminal epithelium, and upregulated the expression of MCT1 in the epithelium of proximal jejunum. In conclusion, heat treatment of CM may negatively affect calf growth and gastrointestinal tract development. Glycerol inclusion may increase starter intake, ADG, ruminal fermentation, and intestinal development in calves when CM is used as a main source of protein in pelleted starter mixture.


Asunto(s)
Alimentación Animal , Brassica napus/metabolismo , Bovinos/metabolismo , Tracto Gastrointestinal/crecimiento & desarrollo , Tracto Gastrointestinal/metabolismo , Glicerol/farmacología , Abomaso/metabolismo , Alimentación Animal/análisis , Animales , Peso Corporal , Bovinos/crecimiento & desarrollo , Dieta/veterinaria , Digestión , Ácidos Grasos Volátiles/metabolismo , Fermentación , Manipulación de Alimentos , Glicerol/metabolismo , Masculino , Rumen/metabolismo , Destete
7.
J Dairy Sci ; 102(10): 8862-8873, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31421880

RESUMEN

The aim of this study was to determine the effect of docosahexaenoic acid-rich algae (DHA-RA) supplementation in milk replacer (MR) on performance, selected cytokine expression in lymphocytes, and blood immunoglobulin concentration in newborn dairy calves. Forty female Holstein-Friesian calves (8.6 ± 0.8 d old and 41.1 ± 4.3 kg; mean ± standard deviation) were blocked by date of birth and allocated into 4 experimental groups (10 animals/group): (1) not supplemented with DHA-RA, (2) supplemented with 9 g of DHA-RA/d in MR, (3) supplemented with 18 g of DHA-RA/d in MR, and (4) supplemented with 27 g of DHA-RA/d in MR. Milk replacer was fed in an amount equal to 900 g of MR powder/d (as fed), 2 times a d, for 49 d. Starter mixture (SM) was fed ad libitum beginning on d 15 of the study. Each calf was in the study over a period of 49 d. The MR and SM intake and fecal score were recorded daily and body weight was recorded weekly. Blood samples were collected before the morning feeding, at the beginning of the study, every consecutive week, and at the end of the study for morphology and smear analysis, serum immunoglobulin level (IgG, IgA, and IgM), and lymphocyte isolation. The mRNA isolated from lymphocytes was checked for TNFα, IL-1ß, IL-6, and p65 expression. Average daily gain between d 1 to 14 of the study increased quadratically with increasing dose of DHA-RA. However, average daily gain between d 15 to 49 of the study tended to linearly decrease and over the whole study linearly decreased with increasing dose of DHA-RA. The MR intake decreased linearly between d 1 to 14 of the study and over the whole study, and mean SM intake decreased quadratically with increasing dose of DHA-RA. Feed efficiency increased quadratically and fecal score decreased quadratically during the first 14 d of the study. Increasing dose of DHA-RA led to cubic changes in feed efficiency and fecal score between d 15 and 49 of the study. Overall, over the whole study period a tendency was observed for lower fecal score for the DHA-RA supplemented groups. Interleukin-1ß mRNA expression decreased linearly, whereas the mRNA expression of p65 and TNFα as well as serum IgG concentration tended to decrease linearly with increasing dose of supplemental DHA-RA. No effect of group was found on IgA and IgM serum level and the majority of blood parameters. Altogether, treatment worsened production variables but seemed to have a beneficial effect on the immune system of calves.


Asunto(s)
Alimentación Animal , Bovinos , Suplementos Dietéticos , Ácidos Docosahexaenoicos/farmacología , Sustitutos de la Leche/farmacología , Estramenopilos , Alimentación Animal/análisis , Animales , Animales Recién Nacidos , Peso Corporal , Dieta/veterinaria , Femenino , Sistema Inmunológico/efectos de los fármacos , Linfocitos/efectos de los fármacos , Linfocitos/metabolismo , Leche , Sustitutos de la Leche/química , Destete
8.
Pol J Vet Sci ; 21(1): 213-216, 2018 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-29624008

RESUMEN

The aim of this study was to determine age-related changes in the mRNA expression of four clusters of differentiation (CD: e.g. CD5, CD21, CD22 and CD23) in lymphocytes of calves. Blood samples were collected from 10 Holstein heifers on day 2, 22 and 56 of life and used for lymphocyte isolation. Subsequently, the mRNA was isolated from lymphocytes and the relative expression of CD5, CD21, CD22 and CD23 was investigated using quantitative real-time PCR with GAPDH as a reference gene. CD5, CD21 and CD23 mRNA expression increased linearly (p ≤ 0.04) with calf age, whereas CD22 mRNA expression did not change in the investigated period (p > 0.05). Age related changes in CD5, CD21 and CD23 mRNA expression suggest their importance in the process of lymphocyte maturation in calves.


Asunto(s)
Envejecimiento/fisiología , Antígenos CD/metabolismo , Bovinos/fisiología , Regulación del Desarrollo de la Expresión Génica/fisiología , ARN Mensajero/metabolismo , Animales , Antígenos CD/genética , Femenino , ARN Mensajero/genética
9.
J Dairy Sci ; 101(6): 4785-4800, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29525310

RESUMEN

Promotion of microbial butyrate production in the reticulorumen is a widely used method for enhancing forestomach development in calves. Additional acceleration of gastrointestinal tract (GIT) development, both the forestomach and lower parts of the GIT (e.g., abomasum, intestine, and also pancreas), can be obtained by dietary butyrate supplementation. For this purpose, different sources (e.g., butyrate salts or butyrins), forms (e.g., protected or unprotected), methods (e.g., in liquid feed or solid feed), and periods (e.g., before or after weaning) of butyrate administration can be used. The aim of this paper was to summarize the knowledge in the field of butyrate supplementation in feeds for newborn calves in practical situations, and to suggest directions of future studies. It has been repeatedly shown that supplementation of unprotected salts of butyrate (primarily sodium salt) in milk replacer (MR) stimulates the rumen, small intestine, and pancreas development in calves, with a supplementation level equating to 0.3% of dry matter being sufficient to exert the desired effect on both GIT development and growth performance. On the other hand, the effect of unprotected butyrins and protected forms of butyrate supplementation in MR has not been extensively investigated, and few studies have documented the effect of butyrate addition into whole milk (WM), with those available focusing mainly on the growth performance of animals. Protected butyrate supplementation at a low level (0.3% of protected product in DM) in solid feed was shown to have a potential to enhance GIT development and performance of calves fed MR during the preweaning period. Justification of this form of butyrate supplementation in solid feed when calves are fed WM or after weaning needs to be documented. After weaning, inclusion of unprotected butyrate salts in solid feed was shown to increase solid feed intake, but the effect on GIT development and function has not been determined in detail, and optimal levels of supplementation are also difficult to recommend based on available reports. Future studies should focus on comparing different sources (e.g., salts vs. esters), forms (e.g., protected vs. unprotected), and doses of supplemental butyrate in liquid feeds and solid feeds and their effect not only on the development of rumen, abomasum, and small intestine but also the omasum and large intestine. Furthermore, the most effective source, form, and dose of supplemental butyrate in solid feed depending on the liquid feed program (e.g., MR or WM), stage of rearing (e.g., pre- or postweaning), and solid composition (e.g., lack or presence of forage in the diet) need to be determined.


Asunto(s)
Ácido Butírico/metabolismo , Bovinos/crecimiento & desarrollo , Tracto Gastrointestinal/crecimiento & desarrollo , Alimentación Animal/análisis , Animales , Bovinos/metabolismo , Dieta/veterinaria , Suplementos Dietéticos/análisis , Tracto Gastrointestinal/metabolismo
10.
J Anim Sci ; 95(5): 2144-2155, 2017 May.
Artículo en Inglés | MEDLINE | ID: mdl-28726987

RESUMEN

The aim of the study was to determine the effect of butyrate infusion into the rumen on butyrate flow to the duodenum, expression of short-chain fatty acid (SCFA) transporters (monocarboxylate transporter-1, -2, and -4) and receptors (G protein coupled receptor-41 and -43) in the duodenal epithelium and nutrient digestion in sheep. Eight wethers (39.0 ± 3.00 kg; mean ± SD) with ruminal and T-shape duodenal cannulas were allocated to 4 × 4 replicated Latin square design with each experimental period lasting for 21 d (12 d of adaptation and 9 d for data and sample collection). Experimental treatments were: 1) distilled water infusion into the rumen (CONT); 2) 15 g/d of butyric acid infusion into the rumen (BUT15); 3) 30 g/d of butyric acid infusion into the rumen (BUT30); and 4) 45 g/d of butyric acid infusion into the rumen (BUT45). The daily dose of butyrate was infused into the rumen via the rumen cannula, with 200 mL of solution of butyric acid and distilled water, at a constant rate (0.1389 mL/min) throughout the day using a peristaltic pump. Correspondingly, 200 mL/d of distilled water was infused into the rumen of CONT. The wethers were fed daily 900 g of chopped meadow hay and 200 g of concentrate in two equal meals at 0600 and 1800 h. Butyrate infusion into the rumen did not affect total SCFA concentration in the rumen fluid ( > 0.11). Molar proportion of butyrate in total SCFA linearly increased, and molar proportion of acetate and isovalerate linearly decreased ( ≤ 0.02) with an increasing amount of butyrate infused into the rumen. The molar proportion of butyrate in total SCFA in the duodenal digesta linearly increased ( < 0.01), and butyrate flow to duodenum tended to linearly increase ( = 0.06) with an increasing dose of exogenous butyrate delivered to the rumen. Butyrate infusion into the rumen did not affect ( ≥ 0.14) the mRNA expression of monocarboxylate transporter-2 and -4 and G protein coupled receptor-43 in the duodenal epithelium. The G protein coupled receptor-41 and monocarboxylate transporter-1 mRNA expression in the duodenal epithelium was very low in many of the analyzed samples. Digestibility of organic matter, neutral detergent fiber, and acid detergent fiber in the stomach (forestomach and abomasum) decreased for BUT15 and BUT30 and then increased for BUT45 (quadratic, ≤ 0.04); however, neither digestibility in the intestine nor total tract digestibility differed between treatments ( ≥ 0.10).


Asunto(s)
Ácido Butírico/farmacología , Ácidos Grasos Volátiles/metabolismo , Ovinos/fisiología , Animales , Fibras de la Dieta/metabolismo , Digestión/efectos de los fármacos , Duodeno/efectos de los fármacos , Duodeno/metabolismo , Epitelio/efectos de los fármacos , Fermentación/efectos de los fármacos , Masculino , Rumen/efectos de los fármacos , Rumen/metabolismo
11.
J Dairy Sci ; 100(8): 6312-6326, 2017 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-28571989

RESUMEN

Fourier transform mid-infrared (FT-MIR) spectra of milk are commonly used for phenotyping of traits of interest through links developed between the traits and milk FT-MIR spectra. Predicted traits are then used in genetic analysis for ultimate phenotypic prediction using a single-trait mixed model that account for cows' circumstances at a given test day. Here, this approach is referred to as indirect prediction (IP). Alternatively, FT-MIR spectral variable can be kept multivariate in the form of factor scores in REML and BLUP analyses. These BLUP predictions, including phenotype (predicted factor scores), were converted to single-trait through calibration outputs; this method is referred to as direct prediction (DP). The main aim of this study was to verify whether mixed modeling of milk spectra in the form of factors scores (DP) gives better prediction of blood ß-hydroxybutyrate (BHB) than the univariate approach (IP). Models to predict blood BHB from milk spectra were also developed. Two data sets that contained milk FT-MIR spectra and other information on Polish dairy cattle were used in this study. Data set 1 (n = 826) also contained BHB measured in blood samples, whereas data set 2 (n = 158,028) did not contain measured blood values. Part of data set 1 was used to calibrate a prediction model (n = 496) and the remaining part of data set 1 (n = 330) was used to validate the calibration models, as well as to evaluate the DP and IP approaches. Dimensions of FT-MIR spectra in data set 2 were reduced either into 5 or 10 factor scores (DP) or into a single trait (IP) with calibration outputs. The REML estimates for these factor scores were found using WOMBAT. The BLUP values and predicted BHB for observations in the validation set were computed using the REML estimates. Blood BHB predicted from milk FT-MIR spectra by both approaches were regressed on reference blood BHB that had not been used in the model development. Coefficients of determination in cross-validation for untransformed blood BHB were from 0.21 to 0.32, whereas that for the log-transformed BHB were from 0.31 to 0.38. The corresponding estimates in validation were from 0.29 to 0.37 and 0.21 to 0.43, respectively, for untransformed and logarithmic BHB. Contrary to expectation, slightly better predictions of BHB were found when univariate variance structure was used (IP) than when multivariate covariance structures were used (DP). Conclusive remarks on the importance of keeping spectral data in multivariate form for prediction of phenotypes may be found in data sets where the trait of interest has strong relationships with spectral variables.


Asunto(s)
Ácido 3-Hidroxibutírico/sangre , Bovinos , Leche/química , Espectroscopía Infrarroja por Transformada de Fourier/veterinaria , Animales , Femenino , Fenotipo , Polonia , Espectroscopía Infrarroja por Transformada de Fourier/métodos
12.
J Dairy Sci ; 100(8): 6298-6311, 2017 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-28571993

RESUMEN

The aim of this study was to estimate genetic parameters for blood ß-hydroxybutyrate (BHB) predicted from milk spectra and for clinical ketosis (KET), and to examine genetic association of blood BHB with KET and milk production traits (milk, fat, protein, and lactose yields, and milk fat, protein, and lactose contents). Data on milk traits, KET, and milk spectra were obtained from the Norwegian Dairy Herd Recording System with legal permission from TINE SA (Ås, Norway), the Norwegian Dairy Association that manages the central database. Data recorded up to 120 d after calving were considered. Blood BHB was predicted from milk spectra using a calibration model developed based on milk spectra and blood BHB measured in Polish dairy cows. The predicted blood BHB was grouped based on days in milk into 4 groups and each group was considered as a trait. The milk components for test-day milk samples were obtained by Fourier transform mid-infrared spectrometer with previously developed calibration equations from Foss (Hillerød, Denmark). Veterinarian-recorded KET data within 15 d before calving to 120 d after calving were used. Data were analyzed using univariate or bivariate linear animal models. Heritability estimates for predicted blood BHB at different stages of lactation were moderate, ranging from 0.250 to 0.365. Heritability estimate for KET from univariate analysis was 0.078, and the corresponding average estimate from bivariate analysis with BHB or milk production traits was 0.002. Genetic correlations between BHB traits were higher for adjacent lactation intervals and decreased as intervals were further apart. Predicted blood BHB at first test day was moderately genetically correlated with KET (0.469) and milk traits (ranged from -0.367 with protein content to 0.277 with milk yield), except for milk fat content from across lactation stages that had near zero genetic correlation with BHB (0.033). These genetic correlations indicate that a lower BHB is genetically associated with higher milk protein and lactose contents, but with lower yields of milk, fat, protein, and lactose, and with lower frequency of KET. Estimates of genetic correlation of KET with milk production traits were from -0.333 (with protein content) to 0.178 (with milk yield). Blood BHB can routinely be predicted from milk spectra analyzed from test-day milk samples, and thereby provides a practical alternative for selecting cows with lower susceptibility to ketosis, even though the correlations are moderate.


Asunto(s)
Ácido 3-Hidroxibutírico/sangre , Enfermedades de los Bovinos/genética , Cetosis/veterinaria , Leche/química , Animales , Bovinos , Enfermedades de los Bovinos/sangre , Dinamarca , Femenino , Cetosis/sangre , Cetosis/genética , Lactancia , Noruega
13.
J Dairy Sci ; 100(6): 4539-4551, 2017 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-28365118

RESUMEN

The aim of this study was to determine the time course for adaptation of the reticulo-rumen, omasum, abomasum, and small intestine in response to an abrupt increase in the proportion of grain in the diet. Adaptive responses include tissue and digesta mass, small intestinal length, and brush border enzyme activity in the duodenum, proximal jejunum, and ileum. Twenty-five Holstein steers (213 ± 23 kg; 5 to 7 mo of age) were blocked by body weight, and within block were randomly assigned to 1 of 5 treatments: the control diet (CTRL; 92% chopped grass hay and 8% mineral and vitamin supplement on a dry matter basis) or a moderate grain diet (MGD; 50% chopped grass hay, 42% rolled barley grain, and 8% mineral and vitamin supplement) that was fed for 3 (MGD3), 7 (MGD7), 14 (MGD14), or 21 d (MGD21). Dry matter intake was limited to 2.25% of body weight to ensure that changes in dry matter intake did not confound the results. On the last day of the dietary exposure, steers were slaughtered 2 h after feeding. Reticulo-rumen tissue mass and ruminal epithelium mass in the ventral sac of the rumen were not affected by the MGD. Wet reticulo-ruminal digesta mass decreased from CTRL to MGD7 and then increased, but reticulo-ruminal digesta dry matter mass did not differ between treatments. Omasal mass, omasal tissue mass, and omasum digesta mass decreased linearly with the number of days fed MGD, but abomasal tissue mass tended to increase linearly. Duodenal tissue mass tended to increase linearly, and ileal length increased linearly with the number of days fed MGD. Lactase activity in the proximal jejunum increased linearly and maltase activity in duodenum tended to increase linearly with days fed MGD. Aminopeptidase N activity in the proximal jejunum increased cubically with days fed MGD, and dipeptidylpeptidase IV activity in ileum tended to decrease from CTRL to MGD14 and then tended to increase. Adaptation to a diet with a greater proportion of concentrate involves changes in the mass and length of regions of the gastrointestinal tract and brush border enzyme activity. These changes take place gradually over at least 3 wk.


Asunto(s)
Adaptación Fisiológica , Grano Comestible/metabolismo , Tracto Gastrointestinal/anatomía & histología , Intestino Delgado/fisiología , Microvellosidades/enzimología , Estómago de Rumiantes/fisiología , Abomaso/anatomía & histología , Abomaso/fisiología , Alimentación Animal , Animales , Dieta , Suplementos Dietéticos , Digestión , Intestino Delgado/anatomía & histología , Intestino Delgado/enzimología , Masculino , Omaso/anatomía & histología , Omaso/fisiología , Poaceae , Rumen/anatomía & histología , Rumen/fisiología , Estómago de Rumiantes/anatomía & histología , Factores de Tiempo , Oligoelementos/administración & dosificación , Vitaminas/administración & dosificación
14.
Pol J Vet Sci ; 19(2): 393-9, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27487515

RESUMEN

The aim of this study was to evaluate the acute phase response of dairy cows to repeated liver biopsy in order to estimate the safety of this procedure during the transition period. Liver biopsies (up to 1000 mg of liver tissue) were conducted twice a day, 7 days before expected parturition and 3 days after calving. The number of needle insertions for each biopsy was recorded and was dependent on the amount of obtained tissue. Blood samples were taken on day 7 before expected parturition, then on days 3, 4, 7 and 14 after calving. Body temperature was measured daily in all 30 cows from day 3 until day 14 after calving. The concentrations of haptoglobin, serum amyloid A, fibrinogen and interleukin-6 were determined in serum and plasma. In 16.7% of cows, the rectal body temperature rose by ≥ 0.5°C on the day after liver biopsy. Although the concentrations of haptoglobin, serum amyloid A and fibrinogen increased significantly after calving (p<0.01), there was no influence of the number of biopsies on the acute phase reaction and repeated biopsy during the transition period had no effect on body temperature. Therefore, the procedure may be regarded as safe for cows during the transition period.


Asunto(s)
Reacción de Fase Aguda/metabolismo , Bovinos/metabolismo , Hígado/patología , Proteínas de Fase Aguda/metabolismo , Animales , Biopsia/efectos adversos , Biopsia/veterinaria , Femenino , Lactancia/fisiología , Embarazo
15.
J Dairy Sci ; 99(2): 1247-1252, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26686714

RESUMEN

The aim of this study was to determine the effect of canola meal use as a protein source in a starter mixture (SM) on feeding behavior and performance of calves during weaning transition. A total of 36 female Holstein calves of a mean age 14.9±1.6 d and body weight 40.1±4.2 kg (mean ± SD) were allocated to 1 of 3 treatments differing in the main source of protein for the SM (12 calves per treatment): (1) soybean meal (TSBM); (2) soybean meal and canola meal (TSBM/TCM); and (3) canola meal (TCM). The SM was offered for ad libitum consumption beginning on the first day of the study, whereas milk replacer (MR) was fed in amounts equal to 900 g (as fed) per day from d 1 to 35 and 450 g/d from d 36 to 42 of the study. Calves were completely weaned on d 43 of the study (57.9±1.6 d of age; mean ± SD), and their performance was monitored for an additional 2 wk. Calf body weight was recorded weekly, and MR and SM intake and fecal fluidity were recorded daily. Feeding behavior of calves during weaning transition, including frequency (no./d), time (min/d), and rate (g/min) of eating the SM as well as frequency and time of drinking water, was monitored on 6 calves per treatment for 2 consecutive days before MR step-down (d 34-35), at MR step-down (d 41-42), and after weaning (d 48-49 of study). Starter mixture intake tended to be higher for TSBM calves as compared with TSBM/TCM calves from d 1 to 35 of the study but was not different between TSBM and TCM calves and was not different between treatments in the whole study period. Calves from TCM treatment had reduced average daily gain (ADG) and feed efficiency (g of ADG/kg of dry matter intake) and a higher fecal score in the period from d 1 to 35 of the study and had lower feed efficiency and tended to have lower ADG in the whole study period as compared with TSBM calves. Average daily gain and feed efficiency did not differ between TSBM and TSBM/TCM calves. Frequency of eating the SM and drinking water as well as time and rate of eating the SM and time of drinking water did not differ between treatments. It is concluded that presence of canola meal in a SM does not affect feeding behavior and performance of calves during weaning transition but has a negative effect on ADG, feed efficiency, and number of days with diarrhea during the preweaning phase of rearing.


Asunto(s)
Alimentación Animal , Brassica napus , Bovinos/fisiología , Proteínas en la Dieta/administración & dosificación , Conducta Alimentaria , Destete , Animales , Peso Corporal , Bovinos/crecimiento & desarrollo , Dieta/veterinaria , Femenino , Leche , Aumento de Peso
16.
J Dairy Sci ; 98(5): 3284-91, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-25726104

RESUMEN

Two trials were conducted to determine the effect of sodium butyrate microencapsulated within triglyceride matrix (Na-butyrate) in the close-up period on performance of dairy cows and rumen papillae development. In trial 1, 26 Holstein-Friesian cows were randomly allocated to 2 groups (13 cows/group) and fed prepartum a total mixed ration (TMR) without or with 300g of Na-butyrate/d from 30 d before expecting calving to parturition. After calving, the same lactational TMR without Na-butyrate was offered to both treatments. Dry matter intake and milk yield were monitored daily to 60 d in milk, and body condition of cows was scored on d 30, 21, and 4 before parturition and d 14, 31, and 60 after parturition. On d 15, 10, and 5 before parturition blood samples were collected from 6 cows randomly chosen from each group and analyzed for plasma ß-hydroxybutyrate and nonesterified fatty acids concentrations. No differences in dry matter (DM) intake, milk yield, body condition score, or plasma ß-hydroxybutyrate and nonesterified fatty acids concentrations was observed between treatments; however, in the last 5 d before parturition the cows receiving Na-butyrate ate 1.7kg of DM/d more, on average, as compared with control cows. In trial 2, 12 Holstein-Friesian growing bulls (404±48; body weight ± SD) were used to determine the effect of Na-butyrate inclusion in the diet on rumen papillae development. Bulls were randomly allocated to 2 groups (6 bulls/group) and fed TMR without or with 2% (on a dry matter basis) of Na-butyrate for 21 d. At the end of the study, bulls were killed and rumen fluid and rumen tissue samples from dorsal and ventral sac of the rumen were collected. No effect of Na-butyrate supplementation on BW of bulls and DMI during the trial period was observed. Sodium butyrate supplementation increased total short-chain fatty acid concentration in the rumen but had no effect on rumen pH, molar proportions of short-chain fatty acids, and NH3-N concentration. In dorsal sac of the rumen, papillae length and papillae cross-section surface area were increased as a result of Na-butyrate supplementation, whereas in the ventral sac a reverse effect was observed (significant treatment × location in the rumen interaction). Both in the dorsal and ventral sac of the rumen, dietary Na-butyrate increased rumen muscle layer thickness. Altogether, results of this study suggest that Na-butyrate supplementation in the close-up diet may have a potential to enhance rumen papillae growth and rumen adaptation to postpartum diet but lactation performance was not affected under conditions of the current study.


Asunto(s)
Alimentación Animal/análisis , Ácido Butírico/administración & dosificación , Bovinos/fisiología , Dieta/veterinaria , Lactancia , Ácido 3-Hidroxibutírico/sangre , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Suplementos Dietéticos , Ácidos Grasos no Esterificados/sangre , Ácidos Grasos Volátiles/metabolismo , Femenino , Concentración de Iones de Hidrógeno , Masculino , Leche/metabolismo , Parto , Periodo Posparto/metabolismo , Rumen/metabolismo , Triglicéridos/metabolismo
17.
J Dairy Sci ; 98(4): 2682-6, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25648817

RESUMEN

The aim of this study was to determine the effect of different inclusion rates of microencapsulated sodium butyrate (M-SB) in the starter mixture (SM) on performance of dairy calves. Forty female Holstein calves with a mean (± SD) age of 12.8 (± 1.5) d were allocated to 1 of 4 treatments (10 calves/treatment) and fed SM without (M-SB-0) or with 0.3% (M-SB-0.3), 0.6% (M-SB-0.6), or 0.9% (M-SB-0.9) of M-SB (as fed) during a 49-d period of milk replacer feeding. The milk replacer was fed at 670 g/d divided into 2 equal meals. Starter mixture with or without M-SB was offered for ad libitum consumption beginning on the first day of the trial. Body weight of calves was recorded weekly, whereas intakes of milk replacer and SM and fecal fluidity were recorded daily. Intake of SM decreased linearly with increasing M-SB inclusion rate. Average daily gain decreased and body weight gain tended to decrease linearly with increasing amounts of M-SB in SM, but feed efficiency was not affected. Fecal score and number of days with diarrhea increased cubically with increasing M-SB inclusion rate in SM. Under the conditions of the current study, supplementation of SM with M-SB had a negative effect on performance of calves.


Asunto(s)
Ácido Butírico/metabolismo , Bovinos/crecimiento & desarrollo , Bovinos/metabolismo , Industria Lechera/métodos , Dieta/veterinaria , Suplementos Dietéticos , Alimentación Animal/análisis , Animales , Ácido Butírico/administración & dosificación , Suplementos Dietéticos/análisis , Metabolismo Energético/fisiología , Femenino , Distribución Aleatoria , Aumento de Peso/fisiología
18.
J Anim Physiol Anim Nutr (Berl) ; 99(3): 521-30, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24980113

RESUMEN

The aim of the present study was to investigate whether besides age and solid feed intake, monocarboxylic acid transporter type 1 (MCT1) expression in the rumen epithelium of calves is affected by liquid feed type [whole milk (WM) or milk replacer (MR)]. Thirty bull calves at the mean age of 5 days were randomly allocated to five experimental groups (six calves/group). Six calves were slaughtered immediately after allocation to the trial (5 days of life), eighteen calves were fed MR and slaughtered at week intervals (on 12, 19, 26 days of life respectively), and six calves were fed WM and slaughtered at the 26 days of life. MCT1 protein abundance and the MCT1 mRNA level were investigated in the dorsal and ventral sack of the rumen. Solid feed intake and short-chain fatty acids (SCFA) concentration in the rumen fluid increased linearly with calves' age. The amount of the MCT1 protein and mRNA in the dorsal sac of rumen as well as the amount of MCT1 protein in the cranial ventral sac of rumen also increased linearly with calves' age. Calves fed WM had greater solid feed intake in the last week of the study as compared to calves fed MR, but SCFA concentration in the rumen fluid was not different. MCT1 mRNA expression in the cranial dorsal sac of rumen and protein MCT1 expression in both dorsal and ventral cranial sack of the rumen were higher in calves fed WM as compared to calves fed MR. This study confirmed age-dependent changes of MCT1 expression in the rumen epithelium of newborn calves and showed that its expression might be affected by liquid feed type.


Asunto(s)
Envejecimiento/fisiología , Animales Recién Nacidos , Epitelio/metabolismo , Regulación del Desarrollo de la Expresión Génica/fisiología , Transportadores de Ácidos Monocarboxílicos/metabolismo , Rumen/metabolismo , Simportadores/metabolismo , Alimentación Animal/análisis , Animales , Bovinos , Masculino , Leche , Sustitutos de la Leche , Transportadores de Ácidos Monocarboxílicos/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Simportadores/genética
19.
J Dairy Sci ; 97(2): 1026-35, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-24342681

RESUMEN

The effect of sodium butyrate (SB) supplementation in milk replacer (MR), starter mixture (SM), or both on small intestine maturation in newborn calves was investigated. Twenty-eight male calves with a mean age of 5 (± 1) d were randomly allocated into 1 of 4 groups (7 animals per group) and fed (1) MR and SM, without SB (MR(-) and SM(-), respectively; MR(-)/SM(-)); (2) MR(-) and SM supplemented with SB encapsulated within triglyceride matrix (SM(+), 0.6% as fed; MR(-)/SM(+)); (3) MR supplemented with crystalline SB (MR(+), 0.3% as fed) and SM(-) (MR(+)/SM(-)); or (4) MR(+) and SM(+) (MR(+)/SM(+)). The MR was offered in amounts equal to 10% of initial body weight of the calf. The SM was blended with whole corn grain (50/50; wt/wt) and offered ad libitum as a starter diet. Calves were slaughtered at 26 d (± 1) of age and small intestine development was investigated. Treatment with MR(+) decreased villus height in the proximal jejunum and decreased villus height, crypt depth, and tunica mucosa thickness in the middle jejunum, whereas treatment with SM(+) tended to increase small intestine weight and crypt depth in the proximal jejunum, and increased villus height in the distal jejunum. In the duodenum, crypt depth and tunica mucosa thickness were greater for the MR(-)/SM(+) group compared with MR(-)/SM(-), MR(+)/SM(-), and MR(+)/SM(+) groups. In the ileum, crypt depth was less for MR(-)/SM(+) compared with MR(-)/SM(-). Supplementation with SB in both MR and SM enhanced cell proliferation and decreased apoptosis in the middle jejunum mucosa. Regarding brush border enzyme activities, addition of SB to MR increased lactase activity in the middle jejunum and maltase activity in the distal jejunum, and tended to increase lactase activity in the distal jejunum, aminopeptidase A activity in the middle jejunum and ileum, and aminopeptidase N activity in the ileum. In contrast, SM(+) increased dipeptidylpeptidase IV activity in the distal jejunum and tended to increase aminopeptidase N in the distal jejunum. In conclusion, both MR(+) and SM(+) affected small intestine development in newborn calves. This effect depended on the method of SB delivery but MR(+) generally had a more pronounced effect. No synergistic effect of SB supplementation into MR and SM was found.


Asunto(s)
Animales Recién Nacidos , Ácido Butírico/farmacología , Bovinos/crecimiento & desarrollo , Intestino Delgado/crecimiento & desarrollo , Animales , Ácido Butírico/administración & dosificación , Bovinos/fisiología , Dieta/veterinaria , Suplementos Dietéticos , Mucosa Intestinal/crecimiento & desarrollo , Intestino Delgado/efectos de los fármacos , Lactasa , Masculino , Leche , Sustitutos de la Leche/química
20.
J Dairy Sci ; 97(2): 911-6, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-24315321

RESUMEN

Measuring gene expression is a commonly used method to monitor the reaction of cells and tissues to changing nutritional or physiological conditions. Selection of appropriate reference genes is a crucial point in gene expression experiments using real-time PCR techniques. Expression of the "ideal" reference gene should not be affected by the experimental treatments or physiological state of the tissue, organ, or the whole organism. Many programs are available from which to choose the most stable reference gene. In this study, 4 algorithms--ΔCt, BestKeeper, NormFinder, and geNorm--were used to assess the expression stability of 5 candidate reference genes: ß-actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ribosomal protein S9 (RPS9), ribosomal protein L32 (RPL32), and TATA-box-binding protein (TBP), for use in an experiment aimed at measuring gene expression in the liver of cows fed glucogenic supplements in the transition from pregnancy to lactation. The results demonstrated that RPS9 and RPL32 were the most stably expressed in the liver under the conditions of the present experiment; the least stably expressed was ACTB.


Asunto(s)
Bovinos/fisiología , Regulación de la Expresión Génica/fisiología , Gluconeogénesis/efectos de los fármacos , Hígado/metabolismo , Algoritmos , Animales , Suplementos Dietéticos , Femenino , Lactancia/genética , Reacción en Cadena en Tiempo Real de la Polimerasa
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