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1.
Anim Sci J ; 91(1): e13473, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33078508

RESUMO

Dairy cows often suffer metabolic disorders due to the challenges of the transition period. The aim of study was to determine the effects of feeding pomegranate by-product silage (PBS) on feed intake and some blood parameters, such as non-esterified fatty acids (NEFA) and beta hydroxy butyric acid (BHBA), interleukin-1 (IL-1), serum amyloid-A (SAA), prostaglandin F2α metabolites (PGFM), and progesterone (P4) in fresh Holstein cows. The experimental groups were as follows: CON (0 g PBS of DM) and PBS (120 g PBS of DM). Results showed that replacing corn silage with PBS had no effect on dry matter intake (DMI) throughout the study. The CON group had highest (p < .05) NEFA at 14 d postpartum and serum BHBA was decreased by feeding PBS, especially on days 21 (p < .05) and 28 (p < .05) and the entire study (p < .01). Cows fed PBS had lower (p < .05) serum Malondialdehyde (MDA) than CON cows at different times. Feeding PBS decreased (p < .01) serum IL-1 compared to CON group (216.7 versus. 515.5 Pg/ml) at 28 d postpartum, and also cows fed 120 g PBS had higher (p < .01) serum P4. It was concluded that feeding PBS had no adverse effect on feed intake, milk production, and some reproductive parameters. Moreover, PBS positively affected serum metabolites in fresh dairy cows.


Assuntos
Fenômenos Fisiológicos da Nutrição Animal/fisiologia , Bovinos/metabolismo , Bovinos/fisiologia , Indústria de Laticínios , Dieta/veterinária , Dinoprosta/metabolismo , Ingestão de Alimentos , Romã (Fruta) , Silagem , Ácido 3-Hidroxibutírico/sangue , Animais , Ácidos Graxos não Esterificados/sangue , Feminino , Inflamação , Interleucina-1/sangue , Lactação , Malondialdeído/sangue , Reprodução
2.
Medicine (Baltimore) ; 99(40): e22465, 2020 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-33019438

RESUMO

The present study aimed to investigate the predictive value of free fatty acid (FFA) in embolic stroke of undetermined source (ESUS) according to the presence of potential embolic sources (PES) after extensive etiologic evaluation.This was a retrospective observational study based on a single-center registry from January 2011 to July 2017. Stroke subtypes were determined through laboratory findings, brain, and angiographic imaging, carotid ultrasonography, transthoracic echocardiography, and 24-hour Holter monitoring. If ESUS was suspected, transesophageal echocardiography was additionally performed. Patients were classified into ESUS with PES and ESUS without PES. PES included mitral annular calcification, mitral valve prolapse, patent foramen ovale, atrial septal aneurysm, spontaneous echo contrast, ventricular aneurysm, and high-risk plaques of aortic arch, or carotid bulb. We compared clinical and laboratory findings between the two groups.Of a total of 110 ESUS patients, 61 patients (55.5%) had no PES. Patients with ESUS without PES had higher levels of serum FFA, systolic blood pressure, diastolic blood pressure (DBP), and left atrial (LA) enlargement compared with those of ESUS with PES. Multivariable analysis demonstrated that the FFA level, DBP, and LA volume index were associated with ESUS without PES [odds ratio (OR) 1.038, 95% confidence interval (CI) 1.019-1.058 for FFA/10 µEq/L, OR 1.414, 95% CI 1.037-1.928 for DBP/10 mm Hg, and OR 1.073, 95% CI 1.009-1.141 for LA volume index].Higher levels of FFA, DBP, and LA volume index are associated with ESUS without PES, highlighting the need to identify the role of these markers in ESUS through further large-scale, multi-center and prospective studies.


Assuntos
Ácidos Graxos não Esterificados/sangue , Embolia Intracraniana/sangue , Acidente Vascular Cerebral/sangue , Idoso , Biomarcadores/sangue , Feminino , Humanos , Embolia Intracraniana/complicações , Embolia Intracraniana/diagnóstico , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Sistema de Registros , Estudos Retrospectivos , Acidente Vascular Cerebral/diagnóstico , Acidente Vascular Cerebral/etiologia
3.
PLoS One ; 15(9): e0239506, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32976523

RESUMO

BACKGROUND: Low carnitine status may underlie the development of insulin resistance and metabolic inflexibility. Intravenous lipid infusion elevates plasma free fatty acid (FFA) concentration and is a model for simulating insulin resistance and metabolic inflexibility in healthy, insulin sensitive volunteers. Here, we hypothesized that co-infusion of L-carnitine may alleviate lipid-induced insulin resistance and metabolic inflexibility. METHODS: In a randomized crossover trial, eight young healthy volunteers underwent hyperinsulinemic-euglycemic clamps (40mU/m2/min) with simultaneous infusion of saline (CON), Intralipid (20%, 90mL/h) (LIPID), or Intralipid (20%, 90mL/h) combined with L-carnitine infusion (28mg/kg) (LIPID+CAR). Ten volunteers were randomized for the intervention arms (CON, LIPID and LIPID+CAR), but two dropped-out during the study. Therefore, eight volunteers participated in all three intervention arms and were included for analysis. RESULTS: L-carnitine infusion elevated plasma free carnitine availability and resulted in a more pronounced increase in plasma acetylcarnitine, short-, medium-, and long-chain acylcarnitines compared to lipid infusion, however no differences in skeletal muscle free carnitine or acetylcarnitine were found. Peripheral insulin sensitivity and metabolic flexibility were blunted upon lipid infusion compared to CON but L-carnitine infusion did not alleviate this. CONCLUSION: Acute L-carnitine infusion could not alleviated lipid-induced insulin resistance and metabolic inflexibility and did not alter skeletal muscle carnitine availability. Possibly, lipid-induced insulin resistance may also have affected carnitine uptake and may have blunted the insulin-induced carnitine storage in muscle. Future studies are needed to investigate this.


Assuntos
Carnitina/administração & dosagem , Ácidos Graxos não Esterificados/sangue , Ácidos Graxos não Esterificados/metabolismo , Resistência à Insulina/fisiologia , Lipídeos/administração & dosagem , Adulto , Carnitina/análogos & derivados , Carnitina/sangue , Estudos Cross-Over , Emulsões/administração & dosagem , Humanos , Bombas de Infusão , Insulina/sangue , Insulina/metabolismo , Masculino , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Fosfolipídeos/administração & dosagem , Óleo de Soja/administração & dosagem , Adulto Jovem
5.
J Dairy Sci ; 103(8): 7431-7450, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32475659

RESUMO

Rations including high amounts of corn silage are currently very common in dairy production. Diets with corn silage as forage source result in a low supply of essential fatty acids, such as α-linolenic acid, and may lead to low conjugated linoleic acid (CLA) production. The present study investigated the effects of abomasal infusion of essential fatty acids, especially α-linolenic acid, and CLA in dairy cows fed a corn silage-based diet on performance, milk composition, including fatty acid (FA) pattern, and lipid metabolism from late to early lactation. Rumen-cannulated Holstein cows (n = 40) were studied from wk 9 antepartum to wk 9 postpartum and dried off 6 wk before calving. The cows were assigned to 1 of 4 treatment groups. Cows were abomasally supplemented with coconut oil (CTRL, 76 g/d), linseed and safflower oil (EFA, 78 and 4 g/d; linseed/safflower oil ratio = 19.5:1; n-6/n-3 FA ratio = 1:3), Lutalin (CLA, 38 g/d; BASF SE, Ludwigshafen, Germany; isomers cis-9,trans-11 and trans-10,cis-12 each 10 g/d) or EFA+CLA. Milk composition was analyzed weekly, and blood samples were taken several times before and after parturition to determine plasma concentrations of metabolites related to lipid metabolism. Liver samples were obtained by biopsy on d 63 and 21 antepartum and on d 1, 28, and 63 postpartum to measure triglyceride concentration. Body composition was determined after slaughter. Supplementation of CLA reduced milk fat concentration, increased body fat mass, and improved energy balance (EB) in late and early lactation, but EB was lowest during late lactation in the EFA group. Cows with CLA treatment alone showed an elevated milk citrate concentration in early lactation, whereas EFA+CLA did not reveal higher milk citrate but did have increased acetone. Milk protein was increased in late lactation but was decreased in wk 1 postpartum in CLA and EFA+CLA. Milk urea was reduced by CLA treatment during the whole period. After calving, the increase of nonesterified fatty acids in plasma was less in CLA groups; liver triglycerides were raised lowest at d 28 in CLA groups. Our data confirm an improved metabolic status with CLA but not with exclusive EFA supplementation during early lactation. Increased milk citrate concentration in CLA cows points to reduced de novo FA synthesis in the mammary gland, but milk citrate was less affected in EFA+CLA cows, indicating that EFA supplementation may influence changes in mammary gland FA metabolism achieved by CLA.


Assuntos
Abomaso , Bovinos/fisiologia , Suplementos Nutricionais , Ácidos Graxos Essenciais/farmacologia , Animais , Composição Corporal/efeitos dos fármacos , Bovinos/sangue , Dieta/veterinária , Ácidos Graxos Essenciais/administração & dosagem , Ácidos Graxos não Esterificados/sangue , Feminino , Lactação/efeitos dos fármacos , Ácidos Linoleicos Conjugados/farmacologia , Leite , Proteínas do Leite/metabolismo , Período Pós-Parto , Gravidez , Rúmen/metabolismo
6.
J Dairy Sci ; 103(8): 7377-7391, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32505402

RESUMO

The main objective of this study was to evaluate the efficacy of intrauterine administration of chitosan microparticles (CM) in curing metritis in dairy cows. A secondary objective was to evaluate the effects of metritis treatments on milk yield, survival, and reproductive performance. Cows with a fetid, watery, red-brownish vaginal discharge were diagnosed with metritis. Holstein cows (n = 826) with metritis from 3 dairies located in northern Florida were blocked by parity (primiparous or multiparous) and, within each block, randomly assigned to one of 3 treatments: CM (n = 276) = intrauterine infusion of 24 g of CM dissolved in 40 mL of sterile distilled water at the time of metritis diagnosis (d 0), 2 (d 2), and 4 (d 4) d later; ceftiofur (CEF; n = 275) = subcutaneous injection of 6.6 mg/kg ceftiofur crystalline-free acid in the base of the ear at d 0 and d 3; Control (CON; n = 275) = no treatment applied at metritis diagnosis. All groups could receive escape therapy if condition worsened. Cure was considered when vaginal discharge became mucoid and not fetid. A group of nonmetritic (NMET; n = 2,436) cows was used for comparison. Data were analyzed by generalized linear mixed and Cox's proportional hazard models. Cows in CM and CON had lesser risk of metritis cure on d 12 than cows in CEF (58.6 ± 5.0 vs. 61.9 ± 4.9% vs. 77.9 ± 3.9, respectively). The proportion of cows culled within 60 days in milk (DIM) was greater for cows in CM than for cows in CEF and CON (21.5 ± 2.7 vs. 9.7 ± 1.9 vs. 11.3 ± 2.0%, respectively). Treatment did not affect rectal temperature or plasma nonesterified fatty acids, ß-hydroxybutyrate, and haptoglobin concentrations. Milk yield in the first 60 DIM differed for all treatments, and it was lowest for CM (35.8 ± 0.3 kg/d), followed by CON (36.8 ± 0.3 kg/d) and CEF (37.9 ± 0.3 kg/d). The hazard of pregnancy up to 300 DIM was lesser for CM than CEF (hazard ratio = 0.62; 95% CI: 0.50-0.76), for CM than CON (hazard ratio = 0.77; 95% CI: 0.62-0.95) and for CON than CEF (hazard ratio = 0.80; 95% CI: 0.65-0.99). Culling was greater, and milk yield and fertility were lesser for CEF than NMET. In summary, CM did not improve the cure of metritis, and was detrimental to milk yield, survival, and fertility compared with CON. In contrast, CEF increased the cure of metritis, milk yield, and fertility compared with CM and CON. Finally, the negative effects of metritis on milk yield culling and fertility could not be completely reversed by CEF.


Assuntos
Doenças dos Bovinos/tratamento farmacológico , Quitosana/uso terapêutico , Endometrite/veterinária , Ácido 3-Hidroxibutírico/sangue , Animais , Antibacterianos/uso terapêutico , Bovinos , Cefalosporinas/uso terapêutico , Quitosana/química , Endometrite/tratamento farmacológico , Ácidos Graxos não Esterificados/sangue , Feminino , Fertilidade/efeitos dos fármacos , Florida , Lactação , Leite , Paridade , Tamanho da Partícula , Gravidez , Reprodução
7.
J Dairy Sci ; 103(8): 7288-7301, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32475669

RESUMO

The objective of this study was to determine how feeding behavior of dairy cows is altered in response to diet-induced negative energy balance, and if this response varies depending on dietary particle size distribution. Multiparous Holstein cows (n = 30; days in milk = 59 ± 5; parity = 2.8 ± 0.19), producing 44.6 ± 1.2 kg/d of milk, were fed (on a dry matter basis) a lactating diet [net energy of lactation (NEL) = 1.66 Mcal/kg; 68% forage, including 1.8% wheat straw] during a 2-wk baseline period. To induce negative energy balance, cows were then exposed for 3 wk to 1 of 2 diets formulated for a 20% reduction in energy available for milk (NEL = 1.58 Mcal/kg; 73% forage, including 17.2% wheat straw). These diets were identical, only varying in straw chop length: (1) long straw diet (LS): straw chopped with a 10.2-cm screen, or (2) short straw diet (SS): straw chopped with a 2.54-cm screen. Cows consumed 25.6 ± 0.26 kg/d during the baseline period. Dry matter intake decreased on the experimental diets; dry matter intake was greater for the SS diet as compared with the LS diet (23.1 vs. 22.5 kg/d; standard error = 0.47). During the baseline period, cow serum nonesterified fatty acids (NEFA) and blood ß-hydroxybutyrate averaged 0.27 ± 0.02 and 0.71 ± 0.05 mmol/L, respectively. During the experimental period, NEFA and ß-hydroxybutyrate averaged 0.34 and 1.04 mmol/L, respectively, with a peak of NEFA (0.63 ± 0.06 mmol/L) occurring 4 d after dietary change. During baseline, cows produced 42.3 ± 0.33 kg/d of milk; milk yield was decreased for both SS cows and LS cows during the experimental period (SS = 39.0, LS = 37.8 kg/d; standard error = 0.67). On the experimental diets, cows spent more time eating (266.8 vs. 221.8 min/d), had longer meal lengths (46.9 vs. 37.5 min/meal), and consumed fewer meals (7.1 vs. 7.7 meals/d) compared with the baseline period. Within the experimental period, LS cows spent more time eating per day than SS cows (LS = 281.3, SS = 252.2 min/d). During the baseline period cows sorted against long particles (>19 mm), did not sort medium particles (8 to 19 mm), and sorted for short (4 to 8 mm) and fine (<4 mm) particles. Cows did not change sorting of long particles on the SS diet, but increased sorting against these on the LS diet. On the SS diet cows did not change their sorting of short and fine particles. On the LS diet cows increased sorting for short and fine particles. In the baseline period, no association was detected between feed sorting and serum NEFA concentration. During the experimental period, greater NEFA concentration was associated with greater sorting in favor of short particles for both the LS and SS diets. Furthermore, greater NEFA concentration was associated with greater sorting against the longest particles for both the LS and SS diets. No associations of blood and meal variables were detected during the experimental period. Overall, cows altered their feed sorting behavior in response to experiencing a diet-induced period of negative energy balance and the severity of negative energy balance was associated with the extent of that change in feed sorting.


Assuntos
Ração Animal/análise , Bovinos/fisiologia , Metabolismo Energético , Comportamento Alimentar , Leite/metabolismo , Ácido 3-Hidroxibutírico/sangue , Animais , Dieta/veterinária , Ácidos Graxos não Esterificados/sangue , Feminino , Lactação , Triticum
8.
Clin Sci (Lond) ; 134(12): 1537-1553, 2020 06 26.
Artigo em Inglês | MEDLINE | ID: mdl-32556103

RESUMO

Hyperuricaemia (HUA) significantly increases the risk of metabolic syndrome and is strongly associated with the increased prevalence of high serum free fatty acids (FFAs) and insulin resistance. However, the underlying mechanisms are not well established, especially the effect of uric acid (UA) on adipose tissue, a vital organ in regulating whole-body energy and FFA homeostasis. In the present study, we noticed that adipocytes from the white adipose tissue of patients with HUA were hypertrophied and had decreased UCP1 expression. To test the effects of UA on adipose tissue, we built both in vitro and in vivo HUA models and elucidated that a high level of UA could induce hypertrophy of adipocytes, inhibit their hyperplasia and reduce their beige-like characteristics. According to mRNA-sequencing analysis, UA significantly decreased the expression of leptin in adipocytes, which was closely related to fatty acid metabolism and the AMPK signalling pathway, as indicated by KEGG pathway analysis. Moreover, lowering UA using benzbromarone (a uricosuric agent) or metformin-induced activation of AMPK expression significantly attenuated UA-induced FFA metabolism impairment and adipose beiging suppression, which subsequently alleviated serum FFA elevation and insulin resistance in HUA mice. Taken together, these observations confirm that UA is involved in the aetiology of metabolic abnormalities in adipose tissue by regulating leptin-AMPK pathway, and metformin could lessen HUA-induced serum FFA elevation and insulin resistance by improving adipose tissue function via AMPK activation. Therefore, metformin could represent a novel treatment strategy for HUA-related metabolic disorders.


Assuntos
Adipócitos/patologia , Tecido Adiposo Bege/patologia , Tecido Adiposo Branco/patologia , Ácidos Graxos não Esterificados/sangue , Hiperuricemia/sangue , Hiperuricemia/tratamento farmacológico , Resistência à Insulina , Metformina/uso terapêutico , Células 3T3-L1 , Adenilato Quinase/metabolismo , Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Tecido Adiposo Bege/efeitos dos fármacos , Tecido Adiposo Branco/efeitos dos fármacos , Adulto , Animais , Ativação Enzimática , Feminino , Humanos , Hipertrofia , Leptina/metabolismo , Lipogênese , Lipólise , Masculino , Metformina/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Transdução de Sinais , Triglicerídeos/metabolismo , Ácido Úrico/sangue
9.
J Chromatogr A ; 1624: 461259, 2020 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-32540084

RESUMO

An analytical procedure for the rapid and selective derivatization of free fatty acids into methyl esters directly in plasma without transmethylation of lipid-bound fatty acids was developed for their analysis by gas chromatography-mass spectrometry. The methyl esters of free fatty acids were obtained by reaction with methyl iodide in the solution of dipolar aprotic solvents and in the presence of solid bases. The mechanism of the methylation reaction with these reagents was investigated. Optimal conditions for the selective methylation of free fatty acids were established using different dipolar aprotic solvents and different solid bases. The possible transmethylation of covalently bonded fatty acids from plasma lipids has been investigated under different experimental conditions in order to be avoided. Total methylation of free fatty acids was achieved in 1 min at room temperature using methyl iodide and anhydrous potassium carbonate or sodium carbonate in dimethyl sulfoxide. Under these conditions, transmethylation of lipid-bound fatty acids was avoided. The methyl esters can be injected directly from the reaction solvents. A plasma volume of 50 µL was used without special purification. The detection limits were around 0.1 ng/µL. The proposed method avoids the drawbacks of the previous methods used for the one-step analysis of individual free fatty acids in human plasma.


Assuntos
Ácidos Graxos não Esterificados/sangue , Cromatografia Gasosa-Espectrometria de Massas/métodos , Humanos , Limite de Detecção , Metilação , Reprodutibilidade dos Testes , Solventes/química , Ácidos Esteáricos/química
10.
J Dairy Sci ; 103(8): 7451-7461, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32448574

RESUMO

Objectives were to evaluate the effects of rumen-protected glucose (RPG) supplementation on milk production, post-absorptive metabolism, and inflammatory biomarkers in transition dairy cows. Fifty-two multiparous cows were blocked by previous 305-d mature-equivalent milk (305ME) yield and randomly assigned to 1 of 2 iso-energetic and iso-nitrogenous treatments: (1) control diet (CON; n = 26) or (2) a diet containing RPG (pre-fresh 5.3% of dry matter and 6.0% of dry matter postpartum; n = 26). Cows received their respective dietary treatments from d -21 to 28 relative to calving, and dry matter intake was calculated daily during the same period. Weekly body weight, milk composition, and fecal pH were recorded until 28 d in milk (DIM), and milk yield was recorded through 105 DIM. Blood samples were collected on d -7, 3, 7, 14, and 28 relative to calving. Data were analyzed using repeated measures in the MIXED procedure (SAS Institute Inc., Cary, NC) with previous 305ME as a covariate. Fecal pH was similar between treatments and decreased (0.6 units) postpartum. Dry matter intake pre- and postpartum were unaffected by treatment, as was milk yield during the first 28 or 105 DIM. Milk fat, protein, and lactose concentration were similar for both treatments. Blood urea nitrogen and plasma glucose concentrations were unaffected by treatment; however, results showed increased concentration of circulating insulin (27%), lower nonesterified fatty acids (28%), and lower postpartum ß-hydroxybutyrate (24%) in RPG-fed cows. Overall, circulating lipopolysaccharide-binding protein and haptoglobin did not differ by treatment, but at 7 DIM, RPG-fed cows had decreased lipopolysaccharide-binding protein and haptoglobin concentrations (31 and 27%, respectively) compared with controls. Supplemental RPG improved some biomarkers of post-absorptive energetics and inflammation during the periparturient period, changes primarily characterized by increased insulin and decreased nonesterified fatty acids concentrations, with a concomitant reduction in acute phase proteins without changing milk production and composition.


Assuntos
Ração Animal , Doenças dos Bovinos/sangue , Glucose/farmacologia , Inflamação/veterinária , Ácido 3-Hidroxibutírico/sangue , Animais , Peso Corporal , Bovinos , Doenças dos Bovinos/tratamento farmacológico , Dieta/veterinária , Suplementos Nutricionais , Ácidos Graxos não Esterificados/sangue , Feminino , Glucose/metabolismo , Inflamação/sangue , Inflamação/tratamento farmacológico , Insulina/sangue , Lactação/efeitos dos fármacos , Lactose/análise , Lactose/metabolismo , Leite , Rúmen/metabolismo
11.
J Dairy Sci ; 103(7): 6354-6363, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32359995

RESUMO

At the beginning of lactation, high-producing cows commonly experience an unbalanced energy status that is often responsible for the onset of metabolic disorders and impaired health and performance. Blood ß-hydroxybutyrate (BHB) and nonesterified fatty acids (NEFA) are indicators of excessive fat mobilization and circulating ketone bodies. Recently, prediction models based on mid-infrared (MIR) spectroscopy have been developed to assess blood BHB and NEFA from routinely collected individual milk samples. This study aimed to estimate genetic parameters of blood BHB and NEFA predicted from milk MIR spectra and to assess their phenotypic and genetic correlations with milk production and composition traits in early-lactation Holstein cows. The data set comprised the first test-day record within lactation and spectra of individual milk samples (n = 22,718) of 13,106 Holstein cows collected from 5 to 35 d in milk (DIM). Blood BHB and NEFA were predicted from milk MIR spectra using previously developed prediction models. Genetic parameters of blood metabolites and milk traits were estimated for the whole observational period (5-35 DIM) and within 6 classes of DIM. Blood BHB and NEFA showed similar genetic variation across DIM, with the highest heritability in the first 10 d after calving (0.31 ± 0.06 and 0.19 ± 0.05 for BHB and NEFA, respectively). The genetic correlation between BHB and NEFA was moderate (0.51 ± 0.05). Genetic correlations of BHB with milk yield, SCS, protein percentage, lactose percentage, and urea nitrogen content were similar to, or at least in the same direction as, the correlations of NEFA with the same traits, whereas opposite correlations were observed with fat percentage and fat-to-protein ratio. Results of the current study suggest that blood BHB and NEFA predicted from milk MIR spectra have genetic variation that is potentially exploitable for breeding purposes. Therefore, they could be used as indicator traits of hyperketonemia in a selection index aimed to reduce the susceptibility of dairy cows to metabolic disorders in early lactation.


Assuntos
Ácido 3-Hidroxibutírico/sangue , Bovinos/sangue , Ácidos Graxos não Esterificados/sangue , Leite/química , Seleção Artificial , Animais , Doenças dos Bovinos/genética , Feminino , Padrões de Herança , Cetose/genética , Cetose/veterinária , Lactação , Lactose/análise , Fenótipo
12.
J Dairy Sci ; 103(7): 6422-6438, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32389474

RESUMO

In high-yielding dairy cattle, severe postpartum negative energy balance is often associated with metabolic and infectious disorders that negatively affect production, fertility, and welfare. Mobilization of adipose tissue associated with negative energy balance is reflected through an increased level of nonesterified fatty acids (NEFA) in the blood plasma. Earlier, identification of negative energy balance through detection of increased blood plasma NEFA concentration required laborious and stressful blood sampling. More recently, attempts have been made to predict blood NEFA concentration from milk samples. In this study, we aimed to develop and validate a model to predict blood plasma NEFA concentration using the milk mid-infrared (MIR) spectra that are routinely measured in the context of milk recording. To this end, blood plasma and milk samples were collected in wk 2, 3, and 20 postpartum for 192 lactations in 3 herds. The blood plasma samples were taken in the morning, and representative milk samples were collected during the morning and evening milk sessions on the same day. To predict plasma NEFA concentration from the milk MIR spectra, partial least squares regression models were trained on part of the observations from the first herd. The models were then thoroughly validated on all other observations of the first herd and on the observations of the 2 independent herds to explore their robustness and wide applicability. The final model could accurately predict blood plasma NEFA concentrations <0.6 mmol/L with a root mean square error of prediction of <0.143 mmol/L. However, for blood plasma with >1.2 mmol/L NEFA, the model clearly underestimated the true level. Additionally, we found that morning blood plasma NEFA levels were predicted with significantly higher accuracy using MIR spectra of evening milk samples compared with MIR spectra of morning samples, with root mean square error of prediction values of, respectively, 0.182 and 0.197 mmol/L, and R2 values of 0.613 and 0.502. These results suggest a time delay between variations in blood plasma NEFA and related milk biomarkers. Based on the MIR spectra of evening milk samples, cows at risk for negative energy status, indicated by detrimental morning blood plasma NEFA levels (>0.6 mmol/L), could be identified with a sensitivity and specificity of, respectively, 0.831 and 0.800. As this model can be applied to millions of historical and future milk MIR spectra, it opens an opportunity for regular metabolic screening and improved resilience phenotyping.


Assuntos
Ácidos Graxos não Esterificados/sangue , Leite/química , Espectrofotometria Infravermelho/veterinária , Ácido 3-Hidroxibutírico/sangue , Animais , Bovinos , Testes Diagnósticos de Rotina , Metabolismo Energético , Ácidos Graxos não Esterificados/química , Feminino , Fertilidade , Humanos , Lactação , Período Pós-Parto , Valor Preditivo dos Testes , Sensibilidade e Especificidade
13.
Nutr Metab Cardiovasc Dis ; 30(7): 1188-1195, 2020 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-32448719

RESUMO

BACKGROUND AND AIMS: Glomerular hyperfiltration (GH) is proposed as one of the earliest events in obesity (OB)-associated renal disease. Children with GH and type-1 diabetes showed increased chemokine levels. Chemokine associations with glomerular filtration rate (GFR) and metabolic features in prepubertal children with overweight (OW)/OB are unknown. METHODS AND RESULTS: Cross-sectional study. 75 prepubertal children (aged: 9.0 ± 1.7 years) with OW/OB were studied. Clinical and metabolic characteristics (including non-esterified fatty acids, NEFA) and GFR (combined Zappitelli equation) were assessed. GH was defined as GFR >135 ml/min.1.73 m2. Serum levels of regulated on activation, normal T cell expressed and secreted (RANTES)/CCL5, interleukin-8 (IL-8)/CXCL8 and monokine-induced by interferon-γ (MIG)/CXCL9 were measured by ELISA. Age- and sex-adjusted correlations and differences were tested. 48% of the cohort was female and 13% were OW, 54% OB and 33% severe OB. Prepubertal children with GH showed lower z-BMI (-12%), NEFA (-26%) and uric acid (-22%) than those without GH (all p < 0.05). Similarly to high sensitivity C-reactive protein (hsCRP), there were no differences in serum chemokines between children with GH or not (all p > 0.05). Adjusted correlations were significant for RANTES and z-BMI (r = 0.26; p < 0.05) and for MIG with z-BMI (r = -0.26; p < 0.05) and with NEFA (r = 0.27; p < 0.05). CONCLUSION: GH was not associated with higher chemokine levels in prepubertal children with OW/OB. Decreased rather than elevated GFR values were correlated with obesity and worse metabolic profiles. Chemokines levels in children with severe OB suggest a regulation of the immune response. Follow-up studies are needed to address the clinical implications of these findings.


Assuntos
Quimiocinas/sangue , Desenvolvimento Infantil , Metabolismo Energético , Taxa de Filtração Glomerular , Nefropatias/fisiopatologia , Rim/fisiopatologia , Obesidade Pediátrica/sangue , Fatores Etários , Biomarcadores/sangue , Criança , Estudos Transversais , Ácidos Graxos não Esterificados/sangue , Feminino , Humanos , Nefropatias/diagnóstico , Nefropatias/epidemiologia , Masculino , Obesidade Pediátrica/complicações , Obesidade Pediátrica/diagnóstico , Obesidade Pediátrica/imunologia , Estudos Prospectivos , Fatores de Risco
14.
J Dairy Sci ; 103(7): 6672-6678, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32331887

RESUMO

The objective of the present study was to use longitudinal data to examine the relationships between blood concentrations of nonesterified fatty acids (NEFA), ß-hydroxybutyrate (BHB), and glucose during the transition period in dairy goats. Weekly blood samples were collected from Saanen goats from a commercial herd in Australia [1-7 yr; body weight 70 ± 16.0 kg; body condition score 2.5 ± 0.3; and daily milk yield 2.4 ± 0.73 L/d; all mean ± standard deviation (SD)]. The weekly prevalence of goats above hyperketonemic levels (BHB ≥0.8 mmol/L) was approximately 6 times greater postpartum than antepartum. As well, of the 935 goats sampled antepartum, 50 (5%) had at least 1 hyperketonemic event, and 823 (88%) had at least 1 event of NEFA above the threshold (≥0.3 mmol/L). Of 847 goats tested postpartum, 258 (30%) had at least 1 hyperketonemic event, and 690 goats (81%) had at least 1 event of NEFA above the threshold (≥ 0.7 mmol/L). Substantial variation was found when analyzing the mean days of maximum NEFA and maximum BHB concentrations antepartum (-11 ± 6.6 and -14 ± 7.2 d, respectively, mean ± SD) and postpartum (14 ± 6.6 and 9 ± 6.8 d, respectively, mean ± SD). We observed moderate to strong relationships between NEFA and BHB concentrations (r = 0.66) and between NEFA and glucose concentrations (r = -0.46) throughout the transition period. Our results suggested that 3 to 16 d in milk is the best sampling window for monitoring hyperketonemia in dairy goats, and that results from simultaneous BHB and glucose tests provide an improved indication of the fat mobilization and energy status of the herd when measured close to this timeframe.


Assuntos
Ácido 3-Hidroxibutírico/sangue , Glicemia/metabolismo , Ácidos Graxos não Esterificados/sangue , Cabras/sangue , Animais , Austrália , Feminino , Doenças das Cabras/sangue , Doenças das Cabras/diagnóstico , Doenças das Cabras/epidemiologia , Cetose/epidemiologia , Cetose/veterinária , Lactação , Estudos Longitudinais , Leite/química , Período Pós-Parto/sangue , Prevalência
15.
J Dairy Sci ; 103(7): 6392-6406, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32331880

RESUMO

The aim of this study was to detect the genomic region or regions associated with metabolic clusters in early-lactation Holstein cows. This study was carried out in 2 experiments. In experiment I, which was carried out on 105 multiparous Holstein cows, animals were classified through k-means clustering on log-transformed and standardized concentrations of blood glucose, insulin-like growth factor I, free fatty acids, and ß-hydroxybutyrate at 14 and 35 d in milk (DIM), into metabolic clusters, either balanced (BAL) or other (OTR). Forty percent of the animals were categorized in the BAL group, and the remainder were categorized as OTR. The cows were genotyped for a total of 777,962 SNP. A genome-wide association study was performed, using a case-control approach through the GEMMA software, accounting for population structure. We found 8 SNP (BTA11, BTA23, and BTAX) associated with the predicted metabolic clusters. In experiment II, carried out on 4,267 second-parity Holstein cows, milk samples collected starting from the first week until 50 DIM were used to determine Fourier-transform mid-infrared (FT-MIR) spectra and subsequently to classify the animals into the same metabolic clusters (BAL vs. OTR). Twenty-eight percent of the animals were categorized in the BAL group, and the remainder were classified in the OTR category. Although daily milk yield was lower in BAL cows, we found no difference in daily fat- and protein-corrected milk yield in cows from the BAL metabolic cluster compared with those in the OTR metabolic cluster. In the next step, a single-step genomic BLUP was used to identify the genomic region(s) associated with the predicted metabolic clusters. The results revealed that prediction of metabolic clusters is a highly polygenic trait regulated by many small-sized effects. The region of 36,258 to 36,295 kb on BTA27 was the highly associated region for the predicted metabolic clusters, with the closest genes to this region (ANK1 and miR-486) being related to hematopoiesis, erythropoiesis, and mammary gland development. The heritability for metabolic clustering was 0.17 (SD 0.03), indicating that the use of FT-MIR spectra in milk to predict metabolic clusters in early-lactation across a large number of cows has satisfactory potential to be included in genetic selection programs for modern dairy cows.


Assuntos
Bovinos/metabolismo , Regulação da Expressão Gênica/fisiologia , Estudo de Associação Genômica Ampla , Lactação/fisiologia , Ácido 3-Hidroxibutírico/sangue , Animais , Glicemia/metabolismo , Estudos de Casos e Controles , Análise por Conglomerados , Ácidos Graxos não Esterificados/sangue , Feminino , Leite/química , Proteínas do Leite/análise , Gravidez
16.
Am J Physiol Renal Physiol ; 318(6): F1418-F1429, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32308019

RESUMO

The use of high dialysate bicarbonate for hemodialysis in end-stage renal disease is associated with increased mortality, but potential physiological mediators are poorly understood. Alkalinization due to high dialysate bicarbonate may stimulate organic acid generation, which could lead to poor outcomes. Using measurements of ß-hydroxybutyrate (BHB) and lactate, we quantified organic anion (OA) balance in two single-arm studies comparing high and low bicarbonate prescriptions. In study 1 (n = 10), patients became alkalemic using 37 meq/L dialysate bicarbonate; in contrast, with the use of 27 meq/L dialysate, net bicarbonate loss occurred and blood bicarbonate decreased. Total OA losses were not higher with 37 meq/L dialysate bicarbonate (50.9 vs. 49.1 meq using 27 meq/L, P = 0.66); serum BHB increased in both treatments similarly (P = 0.27); and blood lactate was only slightly higher with the use of 37 meq/L dialysate (P = 0.048), differing by 0.2 meq/L at the end of hemodialysis. In study 2 (n = 7), patients achieved steady state on two bicarbonate prescriptions: they were significantly more acidemic when dialyzed against a 30 meq/L bicarbonate dialysate compared with 35 meq/L and, as in study 1, became alkalemic when dialyzed against the higher bicarbonate dialysate. OA losses were similar to those in study 1 and again did not differ between treatments (38.9 vs. 43.5 meq, P = 0.42). Finally, free fatty acid levels increased throughout hemodialysis and correlated with the change in serum BHB (r = 0.81, P < 0.001), implicating upregulation of lipolysis as the mechanism for increased ketone production. In conclusion, lowering dialysate bicarbonate does not meaningfully reduce organic acid generation during hemodialysis or modify organic anion losses into dialysate.


Assuntos
Ácido 3-Hidroxibutírico/sangue , Equilíbrio Ácido-Base , Alcalose/sangue , Bicarbonatos/administração & dosagem , Soluções para Hemodiálise/administração & dosagem , Falência Renal Crônica/terapia , Ácido Láctico/sangue , Diálise Renal , Adulto , Idoso , Idoso de 80 Anos ou mais , Alcalose/diagnóstico , Alcalose/etiologia , Alcalose/fisiopatologia , Bicarbonatos/efeitos adversos , Bicarbonatos/metabolismo , Biomarcadores/sangue , Ácidos Graxos não Esterificados/sangue , Feminino , Soluções para Hemodiálise/efeitos adversos , Soluções para Hemodiálise/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Falência Renal Crônica/sangue , Falência Renal Crônica/diagnóstico , Falência Renal Crônica/fisiopatologia , Lipólise , Masculino , Pessoa de Meia-Idade , Diálise Renal/efeitos adversos , Fatores de Tempo , Resultado do Tratamento
17.
J Dairy Sci ; 103(7): 6557-6568, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32331890

RESUMO

Sirtuin 3 (SIRT3), a mitochondrial deacetylase, is a key regulator of energy metabolism in the liver. In nonruminants, the hepatic abundance of SIRT3 is decreased in individuals with nonalcoholic fatty liver diseases, and recovery of SIRT3 alleviates hepatic triacylglycerol (TG) deposition. However, the level of SIRT3 expression and its effects on lipid metabolism in dairy cows have not been characterized. Here we studied the hepatic expression of SIRT3 in cows with fatty liver and the role of SIRT3 in fatty acid metabolism in bovine hepatocytes. This in vivo study involved 10 healthy cows and 10 cows with fatty liver, from which we collected samples of liver tissue and blood. Primary hepatocytes were isolated from Holstein calves and treated with 0, 0.5, or 1.0 mM nonesterified fatty acids (NEFA) for 24 h or transinfected with SIRT3 overexpression adenovirus (Ad-SIRT3)/SIRT3-short interfering (si)RNA for 48 h. Cows with fatty liver displayed lower serum glucose concentrations but higher serum NEFA and ß-hydroxybutyrate concentrations relative to healthy cows. Cows with fatty liver also had significant lower mRNA and protein abundance of hepatic SIRT3. Incubation of primary hepatocytes with NEFA reduced SIRT3 abundance in primary hepatocytes in a dose-dependent manner. Fatty acid (1 mM) treatment also markedly increased the abundance of acetyl-CoA carboxylase 1 (ACC1) and fatty acid synthase (FAS) but significantly decreased the abundance of carnitine palmitoyltransferase I (CPT1A), carnitine palmitoyltransferase II (CPT2), and acyl-CoA oxidase (ACO). Knockdown of SIRT3 by SIRT3-siRNA downregulated the mRNA abundance of CPT1A, CPT2, and ACO. In contrast, overexpression of SIRT3 by Ad-SIRT3 upregulated the mRNA abundance of CPT1A, CPT2, and ACO; downregulated the mRNA abundance of ACC1 and FAS; and consequently, decreased intracellular TG concentrations. Overexpression of SIRT3 ameliorated exogenous NEFA-induced TG accumulation by downregulating the abundance of ACC1 and FAS and upregulating the abundance of CPT1A, CPT2, and ACO in calf hepatocytes. Our data demonstrated that cows with fatty liver had lower hepatic SIRT3 contents, and an increase in SIRT3 abundance by overexpression mitigated TG deposition by modulating the expression of lipid metabolism genes in bovine hepatocytes. These data suggest a possible role of SIRT3 as a therapeutic target for fatty liver disease prevention in periparturient dairy cattle.


Assuntos
Doenças dos Bovinos/metabolismo , Ácidos Graxos não Esterificados/administração & dosagem , Fígado Gorduroso/veterinária , Metabolismo dos Lipídeos/efeitos dos fármacos , Sirtuína 3/metabolismo , Ácido 3-Hidroxibutírico/sangue , Acetil-CoA Carboxilase/efeitos dos fármacos , Acil-CoA Oxidase/efeitos dos fármacos , Animais , Carnitina O-Palmitoiltransferase/efeitos dos fármacos , Bovinos , Doenças dos Bovinos/prevenção & controle , Ácidos Graxos/metabolismo , Ácidos Graxos não Esterificados/sangue , Fígado Gorduroso/metabolismo , Fígado Gorduroso/prevenção & controle , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Mitocôndrias/enzimologia , Sirtuína 3/genética , Triglicerídeos/metabolismo
18.
Metabolism ; 107: 154219, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32240726

RESUMO

BACKGROUND: An adequate metabolic and hormonal response to the switch from rest to exercise is critical for the health benefits of exercise interventions. Previous work suggests that this response is impaired with overweight/obesity but the specific differences between overweight/obese and lean individuals remain unclear. METHODS: We compared glucose and non-esterified fatty acid (NEFA) regulation and the changes of key homeostatic hormones during 45 min of moderate exercise between 17 overweight/obese and 28 lean premenopausal women. For this comparison, we implemented an exercise protocol at 60% of individual peak oxygen uptake, with frequent blood sampling and under fasting conditions. RESULTS: We found that at the same exercise intensity in the overweight/obese and the lean group of women, the metabolic and hormonal response differed. In contrast to the lean group, the overweight/obese group portrayed an activation in the stress axis (adrenocorticotropic hormone (ACTH)/cortisol) and a lower growth hormone (hGH) response and exercise-increase of plasma NEFA. Both groups, however, displayed increased insulin sensitivity during exercise that was accompanied by a normalization of the elevated fasting glucose in the overweight/obese group after 15-20 min. CONCLUSION: We conclude that the response to exercise in overweight/obese subjects indeed differs from that in lean individuals. Additionally, we demonstrate that exercise can elicit beneficial (improved glucose regulation) and unwanted effects (stress axis activation) in overweight/obese subjects at the same time. This second finding suggests that exercise interventions for overweight/obese subjects need careful consideration of intensity and dose in order to achieve the intended results and avoid acute, undesired reactions.


Assuntos
Terapia por Exercício , Exercício Físico , Hormônios/sangue , Obesidade/metabolismo , Obesidade/terapia , Sobrepeso/metabolismo , Sobrepeso/terapia , Hormônio Adrenocorticotrópico/sangue , Adulto , Glicemia/análise , Índice de Massa Corporal , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/terapia , Teste de Esforço , Ácidos Graxos não Esterificados/sangue , Feminino , Hormônio do Crescimento Humano/sangue , Humanos , Insulina/metabolismo , Pré-Menopausa
19.
J Dairy Sci ; 103(6): 5561-5574, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32278565

RESUMO

Inflammation is critical in the progression from benign hepatic lipidosis to pathological hepatic steatosis. The objective of this study was to examine the potential role of the outer mitochondrial membrane protein mitofusin 2 (MFN2) in the etiology of hepatic steatosis in dairy cows during early lactation. Using a nested case-control design, we compared blood and liver samples from 10 healthy cows and 10 age-matched cows with moderate fatty liver. Cows with moderate fatty liver had high liver triacylglycerols, elevated plasma concentrations of free fatty acids (FFA) and ß-hydroxybutyrate, and low concentrations of glucose. Cows with moderate fatty liver had overactivated inflammatory pathways in the liver, as indicated by increased abundance of phosphorylated nuclear factor κB (NF-κB) p65, NLR family pyrin domain containing 3 (NLRP3) and caspase-1 inflammasome protein, and elevated plasma concentrations and hepatic mRNA abundance of their molecular targets IL-1ß, IL-6, and tumor necrosis factor α (TNF-α). In the cell culture model, we were able to replicate our findings in cows with moderate fatty liver: 1.2 mM exogenous FFA decreased the abundance of MFN2 and upregulated phosphorylation levels of the inhibitor of NF-κB (IκB) α and NF-κB p65, the IκB kinase ß activity, and the abundance of NLRP3, caspase-1, IL-1ß, IL-6, and TNF-α. Whereas MFN2 knockdown potentiated the FFA-induced activation of these inflammatory pathways, overexpression of MFN2 attenuated the detrimental effect of excess exogenous FFA by improving mitochondrial function and decreasing the release of reactive oxygen species, suggesting that MFN2 may be a potential therapeutic target for FFA-induced hepatic inflammation in dairy cows during early lactation.


Assuntos
Doenças dos Bovinos/prevenção & controle , Ácidos Graxos não Esterificados/efeitos adversos , Fígado Gorduroso/veterinária , GTP Fosfo-Hidrolases/antagonistas & inibidores , Inflamação/veterinária , Membranas Mitocondriais/metabolismo , Proteínas Mitocondriais/antagonistas & inibidores , Animais , Estudos de Casos e Controles , Bovinos , Ácidos Graxos não Esterificados/sangue , Fígado Gorduroso/induzido quimicamente , Fígado Gorduroso/prevenção & controle , Feminino , GTP Fosfo-Hidrolases/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Inflamação/induzido quimicamente , Inflamação/metabolismo , Lactação/efeitos dos fármacos , Proteínas Mitocondriais/metabolismo , Espécies Reativas de Oxigênio/metabolismo
20.
J Dairy Sci ; 103(6): 5550-5560, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32229128

RESUMO

The efficacy of an electric heat blanket (EHB) has previously been confirmed as an alternative method to evaluate heat stress (HS). However, a pair-feeding design has not been used with the EHB model. Therefore, study objectives were to determine the contribution of the nutritional plane to altered metabolism and productivity during EHB-induced HS. Multiparous Holstein cows (n = 18; 140 ± 10 d in milk) were subjected to 2 experimental periods (P); during P1 (4 d), cows were in thermoneutral conditions with ad libitum feed intake. During P2 (4 d), cows were assigned to 1 of 2 treatments: (1) thermoneutral conditions and pair-fed (PF; n = 8) or (2) EHB-induced HS with ad libitum feed intake (n = 10). Overall, the EHB increased rectal temperature, vaginal temperature, skin temperature, and respiration rate (1.4°C, 1.3°C, 0.8°C, and 42 breaths/min, respectively) relative to PF cows. The EHB reduced dry matter intake (DMI; 47%) and, by design, PF cows had a similar pattern and extent of decreased DMI. Milk yield decreased in EHB and PF cows by 27.3% (12.1 kg) and 13.4% (5.4 kg), respectively, indicating that reduced DMI accounted for only ∼50% of decreased milk synthesis. Milk fat content tended to increase (19%) in the EHB group, whereas in the PF cows it remained similar relative to P1. During P2, milk protein and lactose contents tended to decrease or decreased (1.3 and 2.2%, respectively) in both EHB and PF groups. Milk urea nitrogen remained unchanged in PF controls but increased (34.2%) in EHB cows relative to P1. The EHB decreased blood partial pressure of CO2, total CO2, HCO3, and base excess levels (17, 16, 17, and 81%, respectively) compared with those in PF cows. During P2, the EHB and PF cows had similar decreases (4%) in plasma glucose content, but no differences in circulating insulin were detected. However, a group by day interaction was detected for plasma nonesterified fatty acids; levels progressively increased in PF controls but remained unaltered in the EHB cows. Blood urea nitrogen increased in the EHB cows (61%) compared with the PF controls. In summary, utilizing the EHB model indicated that reduced nutrient intake explains only about 50% of the decrease in milk yield during HS, and the postabsorptive changes in nutrient partitioning are similar to those obtained in climate-controlled chamber studies. Consequently, the EHB is a reasonable and economically feasible model to study environmental physiology of dairy cows.


Assuntos
Bovinos/fisiologia , Ingestão de Alimentos , Ácidos Graxos não Esterificados/sangue , Resposta ao Choque Térmico , Proteínas do Leite/metabolismo , Leite/metabolismo , Animais , Roupas de Cama, Mesa e Banho/veterinária , Nitrogênio da Ureia Sanguínea , Dieta/veterinária , Feminino , Temperatura Alta , Insulina/sangue , Lactação , Leite/química , Proteínas do Leite/química , Taxa Respiratória
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