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INTRODUCTION: Relative oxidation of different metabolic substrates in the heart varies both physiologically and pathologically, in order to meet metabolic demands under different circumstances. 13C labelled substrates have become a key tool for studying substrate use-yet an accurate model is required to analyse the complex data produced as these substrates become incorporated into the Krebs cycle. OBJECTIVES: We aimed to generate a network model for the quantitative analysis of Krebs cycle intermediate isotopologue distributions measured by mass spectrometry, to determine the 13C labelled proportion of acetyl-CoA entering the Krebs cycle. METHODS: A model was generated, and validated ex vivo using isotopic distributions measured from isolated hearts perfused with buffer containing 11 mM glucose in total, with varying fractions of universally labelled with 13C. The model was then employed to determine the relative oxidation of glucose and triacylglycerol by hearts perfused with 11 mM glucose and 0.4 mM equivalent Intralipid (a triacylglycerol mixture). RESULTS: The contribution of glucose to Krebs cycle oxidation was measured to be 79.1 ± 0.9%, independent of the fraction of buffer glucose which was U-13C labelled, or of which Krebs cycle intermediate was assessed. In the presence of Intralipid, glucose and triglyceride were determined to contribute 58 ± 3.6% and 35.6 ± 0.8% of acetyl-CoA entering the Krebs cycle, respectively. CONCLUSION: These results demonstrate the accuracy of a functional model of Krebs cycle metabolism, which can allow quantitative determination of the effects of therapeutics and pathology on cardiac substrate metabolism.
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Mitocôndrias/metabolismo , Miocárdio/metabolismo , Acetilcoenzima A/análise , Animais , Isótopos de Carbono , Ciclo do Ácido Cítrico/fisiologia , Glucose/metabolismo , Coração/fisiologia , Masculino , Espectrometria de Massas/métodos , Modelos Biológicos , Oxirredução , Ratos , Ratos WistarRESUMO
Active glycolysis and glutaminolysis provide bioenergetic stability of cancer cells in physiological conditions. Under hypoxia, metabolic and mitochondrial disorders, or pharmacological treatment, a deficit of key metabolic substrates may become life-threatening to cancer cells. We analysed the effects of mitochondrial uncoupling by FCCP on the respiration of cells fed by different combinations of Glc, Gal, Gln and Pyr. In cancer PC12 and HCT116 cells, a large increase in O2 consumption rate (OCR) upon uncoupling was only seen when Gln was combined with either Glc or Pyr. Inhibition of glutaminolysis with BPTES abolished this effect. Despite the key role of Gln, addition of FCCP inhibited respiration and induced apoptosis in cells supplied with Gln alone or Gal/Gln. For all substrate combinations, amplitude of respiratory responses to FCCP did not correlate with Akt, Erk and AMPK phosphorylation, cellular ATP, and resting OCR, mitochondrial Ca(2+) or membrane potential. However, we propose that proton motive force could modulate respiratory response to FCCP by regulating mitochondrial transport of Gln and Pyr, which decreases upon mitochondrial depolarisation. As a result, an increase in respiration upon uncoupling is abolished in cells, deprived of Gln or Pyr (Glc). Unlike PC12 or HCT116 cells, mouse embryonic fibroblasts were capable of generating pronounced response to FCCP when deprived of Gln, thus exhibiting lower dependence on glutaminolysis. Overall, the differential regulation of the respiratory response to FCCP by metabolic environment suggests that mitochondrial uncoupling has a potential for substrate-specific inhibition of cell function, and can be explored for selective cancer treatment.
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Carbonil Cianeto p-Trifluormetoxifenil Hidrazona/metabolismo , Metabolismo Energético , Mitocôndrias/metabolismo , Neoplasias/metabolismo , Consumo de Oxigênio/fisiologia , Animais , Apoptose/genética , Carbonil Cianeto p-Trifluormetoxifenil Hidrazona/química , Respiração Celular/fisiologia , Galactose/metabolismo , Glucose/metabolismo , Glutamina/metabolismo , Glicólise/genética , Células HCT116 , Humanos , Camundongos , Neoplasias/patologia , Fosforilação Oxidativa , Células PC12 , Ácido Pirúvico/metabolismo , Ratos , Especificidade por SubstratoRESUMO
BACKGROUND: Mammalian spermatozoa need to undergo a process named capacitation to be able to fertilize an oocyte. During their journey in the female tract, spermatozoa obtain energy while exposed to a changing environment containing a variety of metabolic substrates. The energy requirements for sperm capacitation are species-specific. In addition, the available energy source can hinder the process of sperm capacitation and eventually the acrosome reaction. OBJECTIVES: To evaluate whether the metabolic substrates available in the in vitro sperm capacitation medium allow or interfere with the pig sperm capacitation process. MATERIAL AND METHODS: The effect of different metabolic substrates on sperm capacitation process was evaluated by analyzing phosphorylation in the p32 protein; the acrosome reaction and the ATP intracellular content. RESULTS: The presence of glucose in the in vitro capacitating medium diminishes, in a concentration-dependent manner, parameters associated with the capacitated status: induced acrosome exocytosis, plasma membrane destabilization, and protein tyrosine phosphorylation. Conversely, sperm incubation with pyruvate or lactate, either individually or in combination, allows the attainment of the capacitated status. Unexpectedly, pig spermatozoa incubated without any extracellular energy substrates or with a non-metabolizable substrate (l-glucose) for 4 h displayed similar sperm viability to the control and exhibited a capacitated phenotype. The capacitation-like phenotype observed in starved pig spermatozoa (absence of glucose, lactate, and pyruvate) was dependent on extracellular bicarbonate and calcium levels, and these spermatozoa exhibited lower intracellular ATP content compared to those not capacitated. Nevertheless, the intracellular content of calcium was not modified in comparison to the control. DISCUSSION AND CONCLUSIONS: Our findings suggest that the metabolic substrates used to fuel pig sperm metabolism are important in achieving the capacitated status. The results of this work could be used to refine the capacitating medium employed in pig in vitro fertilization.
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Background: The early attack of coronary heart disease (CHD) is very hidden, and clinical symptoms generally do not appear until cardiovascular events occur. Therefore, an innovative method is needed to judge the risk of cardiovascular events and guide clinical decision conveniently and sensitively. The purpose of this study is to find out the risk factors related to MACE during hospitalization. In order to develop and verify the prediction model of energy metabolism substrates, and establish a nomogram to predict the incidence of MACE during hospitalization and evaluate their performance. Methods: The data were collected from the medical record data of Guang'anmen Hospital. This review study was collected the comprehensive clinical data of 5,935 adult patients hospitalized in the cardiovascular department from 2016 to 2021. The outcome index was the MACE during hospitalization. According to the occurrence of MACE during hospitalization, these data were divided into MACE group (n = 2,603) and non-MACE group (n = 425). Logistic regression was used to screen risk factors, and establish the nomogram to predict the risk of MACE during hospitalization. Calibration curve, C index and decision curve were used to evaluate the prediction model, and drawn ROC curve to find the best boundary value of risk factors. Results: The logistic regression model was used to establish a risk model. Univariate logistic regression model was mainly used to screen the factors significantly related to MACE during hospitalization in the training set (each variable is put into the model in turn). According to the factors with statistical significance in univariate logistic regression, five cardiac energy metabolism risk factors, including age, albumin(ALB), free fatty acid(FFA), glucose(GLU) and apolipoprotein A1(ApoA1), were finally input into the multivariate logistic regression model as the risk model, and their nomogram were drawn. The sample size of the training set was 2,120, the sample size of the validation set was 908. The C index of the training set is 0.655 [0.621,0.689], and the C index of the validation set was 0.674 [0.623,0.724]. The calibration curve and clinical decision curve show that the model performs well. The ROC curve was used to establish the best boundary value of the five risk factors, which could quantitatively present the changes of cardiac energy metabolism substrate, and finally achieved prediction of MACE during hospitalization conveniently and sensitively. Conclusion: Age, albumin, free fatty acid, glucose and apolipoprotein A1 are independent factors of CHD in MACE during hospitalization. The nomogram based on the above factors of myocardial energy metabolism substrate provides prognosis prediction accurately.
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Although the preferred cardiac metabolic fuels are fatty acids, glucose metabolism also plays an important role. However, irrespective of substrate type, energy generation results in mitochondrial reactive oxygen species (ROS) formation. To determine if the preference of fat over carbohydrates predisposes cardiomyocytes to oxidant production, we measured total and site-specific H2O2 emission in heart mitochondria oxidizing palmitoylcarnitine or pyruvate during copper (Cu) exposure. H2O2 emission was higher during oxidation of palmitoylcarnitine compared with pyruvate. Moreover, the bulk of the H2O2 emitted during palmitoylcarnitine oxidation originated from the outer ubiquinone binding site of complex III (site IIIQo) and the flavin site of electron transfer flavoprotein (site EF). We found no evidence of ROS production from complex I ubiquinone-binding site (site IQ) by reverse electron transport during oxidation of palmitoylcarnitine. Pyruvate oxidation also drove H2O2 emission primarily from sites IIIQo; however, the flavin sites of pyruvate dehydrogenase (site PF) and complex II (site IIF) contributed substantially. The effect of Cu depended on substrate and redox site, with effects at sites OF and IIIQo being more pronounced in mitochondria oxidizing pyruvate compared with palmitoylcarnitine. Cu imposed a concentration-saturable effect at site PF but concentration-dependently stimulated H2O2 emission at site EF. The substrate-dependent differences in H2O2 emission and effects of Cu suggest that fuel type and points of entry of electrons into the mitochondrial electron transport system determine the mitochondrial ROS production rate. Importantly, knowledge of sites of mitochondrial ROS production is crucial to the understanding of cardiac dysfunction associated with impaired substrate metabolism.
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Cobre , Peróxido de Hidrogênio , Cobre/metabolismo , Ácidos Graxos/metabolismo , Peróxido de Hidrogênio/metabolismo , Mitocôndrias Cardíacas , Oxirredução , Piruvatos/farmacologia , Espécies Reativas de Oxigênio/metabolismoRESUMO
Human cytochrome P450 enzyme 1A2 (CYP1A2) is one of the most important cytochrome P450 (CYP) enzymes in the liver, accounting for 13% to 15% of hepatic CYP enzymes. CYP1A2 metabolises many clinical drugs, such as phenacetin, caffeine, clozapine, tacrine, propranolol, and mexiletine. CYP1A2 also metabolises certain precarcinogens such as aflatoxins, mycotoxins, nitrosamines, and endogenous substances such as steroids. The regulation of CYP1A2 is influenced by many factors. The transcription of CYP1A2 involves not only the aromatic hydrocarbon receptor pathway but also many additional transcription factors, and CYP1A2 expression may be affected by transcription coactivators and compression factors. Degradation of CYP1A2 mRNA and protein, alternative splicing, RNA stability, regulatory microRNAs, and DNA methylation are also known to affect the regulation of CYP1A2. Many factors can lead to changes in the activity of CYP1A2. Smoking, polycyclic aromatic hydrocarbon ingestion, and certain drugs (e.g., omeprazole) increase its activity, while many clinical drugs such as theophylline, fluvoxamine, quinolone antibiotics, verapamil, cimetidine, and oral contraceptives can inhibit CYP1A2 activity. Here, we review the drugs metabolised by CYP1A2, the metabolic mechanism of CYP1A2, and various factors that influence CYP1A2 metabolism. The metabolic mechanism of CYP1A2 is of great significance in the development of personalised medicine and CYP1A2 target-based drugs.
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Inibidores do Citocromo P-450 CYP1A2/farmacologia , Citocromo P-450 CYP1A2/metabolismo , Animais , Inibidores Enzimáticos/farmacologia , HumanosRESUMO
Di-(2-ethylhexyl) phthalate (DEHP) is a chemical plasticizer that has been commonly used in the manufacture of polyvinyl chloride. DEHP is one of the environmental pollution sources. In this study, a gram-negative strain RX bacterium utilizing DEHP as sole carbon source was isolated from activated sludge through screening test. This strain RX was identified as Achromobacter sp. RX based on its morphology, physiological properties and 16S rRNA gene sequence analysis. The results showed that the optimal conditions for the DEHP degradation were 30.0 °C and pH 7.0. The DEHP degradation induced by strain RX demonstrated nitrogen source dependent, while followed a decreasing degradation rate under the source of: NO3- > NH4+ > NO2-. The biodegradability of Achromobacter sp. RX was enhanced with Masson pine seed powder as a co-metabolic substrate and Tween-80 as a solubilizing agent. Meanwhile, the degrading kinetics analysis was performed in the condition of DEHP as sole carbon source. The DEHP degradation curves fitted well with the first-order kinetic model at 50-300 mg/L of DEHP, with the half-life ranging from 13.0 to 16.4 h. During the biodegradation of DEHP, mono-(2-ehtylhexyl) phthalate (MEHP) was firstly generated through de-esterification, followed by the formation of phthalic acid and benzoic acid after further de-esterification of MEHP. Benzoic acid was finally mineralized to CO2 and H2O. The decontamination of DEHP-contaminated soil by Achromobacter sp. RX was investigated using a rotating-drum bioreactor. Evolution of total organic carbon from the contaminated soil showed that 86.4%-91.7% of DEHP was mineralized at pH 7.0 and 30.0 °C within 96 h. Reusability of Achromobacter sp. RX and its lifetime were observed over six consecutive cycles. Thus, Achromobacter sp. RX possessed high DEHP biodegradability, which provided a good potential in dealing with DEHP-contaminated soil.
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Achromobacter , Dietilexilftalato , Ácidos Ftálicos , Biodegradação Ambiental , RNA Ribossômico 16S/genéticaRESUMO
Background This study was performed to characterize the metabolic, functional, and structural cardiac changes in a canine model of radiation-induced heart disease by serial in vivo imaging and ex vivo analyses. Methods and Results Thirty-six dogs were randomly assigned to control or irradiated groups at 3 time points (months 3, 6, and 12 after radiation; each group comprised 6 dogs). The left anterior myocardium of dogs in irradiated groups was irradiated locally with a single dose of 20-Gy X-ray. The irradiated myocardial regions showed increased myocardial uptake of 18F-FDG (18F-fludeoxyglucose) in the irradiated beagles, but the increased uptake area decreased at months 6 and 12 compared with month 3 after radiation. Abnormality of myocardial perfusion and cardiac function were detected at month 6 after radiation. Compared with the control groups, the protein expression of GLUT4 (glucose transporter 4) was upregulated in the irradiated groups, correlating with significantly decreased CPT1 (carnitine acyltransferase 1) expression. Mitochondria degeneration, swelling, and count reduction in the irradiated groups were observed. The difference in CD68 of macrophage markers and the inflammatory cytokines (IL-6 [interleukin 6], TNF-α [tumor necrosis factor α]) between the irradiation and control groups was not significant. Furthermore, the progressive aggravation of apoptosis and fibrosis was displayed. Conclusions Elevated 18F-FDG uptake occurred after irradiation and subsequently led to ventricular perfusion defects and dysfunction. The process was associated with myocardial metabolic substrate remodeling, cardiac muscle cell apoptosis, and myocardial fibrosis rather than inflammation.
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Doenças do Cão/metabolismo , Coração/efeitos da radiação , Miocárdio/metabolismo , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Remodelação Ventricular/efeitos da radiação , Amônia , Animais , Western Blotting , Doenças do Cão/diagnóstico por imagem , Cães , Fluordesoxiglucose F18 , Glucose/metabolismo , Masculino , Radioisótopos de Nitrogênio , Reação em Cadeia da Polimerase em Tempo RealRESUMO
3-Iodothyronamine (T1AM) is a structural analog of thyroid hormone that has been demonstrated to have potent affects on numerous physiological systems. Most studies on T1AM have explored its effects in healthy functional systems; while its potential therapeutic uses and safety, and efficacy in pathological conditions are largely unknown. We sought to evaluate the effects of T1AM and its structural analog SG-2 on cancer cell growth and viability. We analyzed the cytotoxicity of these analogs on MCF7 breast cancer cells, HepG2 hepatocellular cancer cells as well as normal control cells using primary human foreskin fibroblasts and mouse preadipocytes control cells. The cytotoxicity of T1AM and SG-2 was determined by cell growth curves, and validated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell viability assays. Cellular uptake analysis was conducted using confocal microscopy. Real-time (RT)-PCR was conducted to identify gene pathways affected by SG-2 in cancer cells. The IC 50 of T1AM was approximately double the concentration of its analog SG-2 in cancer cells. Cytotoxicity studies on normal cells revealed that IC 50 concentrations of SG-2 in cancer cells had no significant impact on cell viability in these cell types. Cell-imaging experiments demonstrated rapid uptake and localization to the mitochondrial membrane. T1AM and SG-2 are able to reduce cancer cell growth and viability. These findings support the potential for use of these compounds and related analogs for their antiproliferation properties in cancer cells.
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Aldehyde dehydrogenases (ALDHs), as essential regulators of aldehyde metabolism in the human body, protect organisms from damage induced by active aldehydes. Given their roles in different cancer types, ALDHs have been evaluated as potential prognostic markers of cancer. ALDHs exhibit high activity in cancer stem cells (CSCs) and may serve as markers of CSCs. Moreover, studies indicated that ALDHs and their regulated retinoic acid, reactive oxygen species and reactive aldehydes metabolism were strongly related with various properties of CSCs. Besides, recent research evidences have demonstrated the transcriptional and post-translational regulation of ALDH expression and activation in CSCs. Thus, this review focuses on the function and regulation of ALDHs in CSCs, particularly ALDH1A1 and ALDH1A3.
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Aldeído Desidrogenase/fisiologia , Neoplasias/enzimologia , Células-Tronco Neoplásicas/enzimologia , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Resistencia a Medicamentos Antineoplásicos , Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , Transdução de SinaisRESUMO
BACKGROUND: Caffeine (CAF) has been shown to improve performance during early phase of repeated sprint exercise; however some studies show that CAF also increases the magnitude of physical stress represented by augmented blood lactate, glucose, and cortisol concentrations during latter phase of repeated sprint exercise. No studies have investigated the efficacy of combined carbohydrate (CHO) and CAF consumption during repeated sprint exercise (RSE) in female athletes. Thus, the purpose of this study was to investigate the effects of CAF with CHO supplementation on RSE and agility. METHODS: Eleven female athletes completed four experimental trials performed 7 d apart in a double-blind, randomized, and counter-balanced crossover design. Treatments included CAF + PLA (placebo), CAF + CHO, PLA + CHO, and PLA + PLA. Participants ingested capsules containing 6 mg · kg(-1) of CAF or PLA 60-min prior to RSE, and 0.8 g · kg(-1) of CHO solution or PLA immediately before the RSE, which consisted of ten sets of 5 × 4-s sprints on the cycle ergometer with 20-s active recovery. The agility T-test (AT-test) was performed before and after the RSE. Blood samples were acquired to assess glucose, lactate, testosterone, and cortisol. RESULTS: During Set 6 of RSE, peak power and mean power were significantly higher in PLA + CHO than those in CAF + PLA and PLA + PLA, respectively (p < .05). Total work was significantly increased by 4.8% and 5.9% with PLA + CHO than those of CAF + CHO and CAF + PLA during Set 3. PLA + CHO also increased total work more than CAF + PLA and PLA + PLA did during Set 6 (p < .05). No significant differences in AT-test performance either before or after the RSE were occurred among treatments (p > .05). Blood lactate and glucose concentrations were significantly higher under CAF + CHO, CAF + PLA, and PLA + CHO versus PLA + PLA (p < .05), but no differences in testosterone or cortisol levels were found (p > .05). CONCLUSIONS: Findings indicate that CAF + PLA or CAF + CHO ingestion did not improve repeated sprint performance with short rest intervals or agility. However, CHO ingested immediately prior to exercise provided a small but significant benefit on RSE performance in female athletes.
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This study aimed at analyzing the energetic substrate (ES) in the main storage tissues of Steindachneridion parahybae, throughout the reproductive cycle in captivity. Differently from wild, in captivity, feeding is not interrupted during the reproductive period, the females do not spawn spontaneously, and they are sedentary. Adult females were sampled monthly and based on their histology and gonadosomatic index (GSI), ovaries were classified into: previtellogenic (PRV), vitellogenic (VTG), and regression (REG) stages. Ovaries at the VTG stage showed higher protein and lipids levels than at the PRV stage with a positive correlation between these substrates and the GSI. Muscle was the main source of proteins transferred to the ovaries, according to the negative correlation between these organs. Lipids remained unchanged in the liver, which is an important supplier in vitellogenesis, a pattern that probably occurs due to the continuous feeding. Muscular glycogen levels were higher at the VTG and REG than at the PRV stages. Plasma triglycerides were also higher during REG, while glucose levels were more elevated during the VTG stage. These results suggest that with constant food supply, the pattern of deposition of ES in S. parahybae is different from that described for other wild potamodromous species.(AU)
O objetivo deste estudo foi analisar a composição do substrato energético (SE) nos principais tecidos de armazenamento de Steindachneridion parahybae, durante todo o ciclo reprodutivo em cativeiro. Diferentemente do ambiente natural, em cativeiro, a alimentação desses animais não é interrompida durante o período reprodutivo, as fêmeas não desovam espontaneamente, e são sedentárias. Fêmeas adultas foram amostradas mensalmente e baseada na histologia e no índice gonadossomático (IGS), os ovários foram classificados: estádios pré-vitelogênico (PRV), vitelogênico (VTG) e regressão (REG). Os ovários no estádio VTG apresentaram uma maior concentração de lipídeos e proteínas em relação ao estágio PRV. Esses substratos correlacionaram-se positivamente com o IGS. O músculo foi a principal fonte de proteína transferida aos ovários, como foi confirmado pela análise de correlação negativa entre esses órgãos. Os lipídeos mantiveram-se inalterados no fígado, considerado um importante órgão fornecedor de lipídeos para a vitelogênese, padrão que possivelmente ocorreu devido à contínua alimentação. A concentração do glicogênio muscular foi mais elevada durante os estágios VTG e REG em relação ao PRV. A concentração de triglicerídeos plasmática apresentou maiores valores no estádio REG enquanto a concentração de glicose no plasma foi maior durante os estádios VTG. Esses resultados sugerem que com alimentação constante, as fêmeas de S. parahybae apresentam um distinto padrão de mobilização dos substratos energéticos em relação ao que já foi descrito para outras espécies potamódromas de ambiente natural.(AU)
This study aimed at analyzing the energetic substrate (ES) in the main storage tissues of Steindachneridion parahybae, throughout the reproductive cycle in captivity. Differently from wild, in captivity, feeding is not interrupted during the reproductive period, the females do not spawn spontaneously, and they are sedentary. Adult females were sampled monthly and based on their histology and gonadosomatic index (GSI), ovaries were classified into: previtellogenic (PRV), vitellogenic (VTG), and regression (REG) stages. Ovaries at the VTG stage showed higher protein and lipids levels than at the PRV stage with a positive correlation between these substrates and the GSI. Muscle was the main source of proteins transferred to the ovaries, according to the negative correlation between these organs. Lipids remained unchanged in the liver, which is an important supplier in vitellogenesis, a pattern that probably occurs due to the continuous feeding. Muscular glycogen levels were higher at the VTG and REG than at the PRV stages. Plasma triglycerides were also higher during REG, while glucose levels were more elevated during the VTG stage. These results suggest that with constant food supply, the pattern of deposition of ES in S. parahybae is different from that described for other wild potamodromous species.(AU)
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Animais , Peixes-Gato/metabolismo , Substratos para Tratamento Biológico/análise , Ração Animal/análise , GlicogênioRESUMO
Fluctuating environmental conditions can promote diversity and control dominance in community composition. In addition to seasonal temperature and moisture changes, seasonal supply of metabolic substrates selects populations temporally. Here we demonstrate cascading effects in the supply of metabolic substrates on methanogenesis and community composition of anaerobic methanogenic archaea in three contrasting peatlands in upstate New York. Fresh samples of peat soils, collected about every 3 months for 20 months and incubated at 22 ± 2°C regardless of the in situ temperature, exhibited potential rates of methane (CH(4)) production of 0.02-0.2 mmol L(-1) day(-1) [380-3800 nmol g(-1) (dry) day(-1)). The addition of acetate stimulated rates of CH(4) production in a fen peatland soil, whereas addition of hydrogen (H(2)), and simultaneous inhibition of H(2)-consuming acetogenic bacteria with rifampicin, stimulated CH(4) production in two acidic bog soils, especially, in autumn and winter. The methanogenic community structure was characterized using T-RFLP analyses of SSU rRNA genes. The E2 group of methanogens (Methanoregulaceae) dominated in the two acidic bog peatlands with relatively greater abundance in winter. In the fen peatland, the E1 group (Methanoregulaceae) and members of the Methanosaetaceae were co-dominant, with E1 having a high relative abundance in spring. Change in relative abundance profiles among methanogenic groups in response to added metabolic substrates was as predicted. The acetate-amendment increased abundance of Methanosarcinaceae, and H(2)-amendment enhanced abundance of E2 group in all peat soils studied, respectively. Additionally, addition of acetate increased abundance of Methanosaetaceae only in the bog soils. Variation in the supply of metabolic substrates helps explain the moderate diversity of methanogens in peatlands.