RESUMO
BACKGROUND: The methodological procedures for studying the fibrolytic activity of rumen bacteria are not clearly established. In this study the efficiency of sonication treatment and buffer composition (i.e. buffer varying in tonicity or pH) on the level of protein extraction from the residue of forage samples incubated in the rumen of a grazing steer and the effect of buffer composition or CaCl2 concentration on the carboxymethylcellulase (CMCase) activity of the released protein were evaluated. RESULTS: The amount of protein released from the residue of incubation was higher (P < 0.05) for the sonicated material and increased linearly with increasing buffer pH (P < 0.05). The CMCase activity of the released protein was not improved by sonication treatment, whereas it was higher (P < 0.05) for hypotonic than for hypertonic buffer. Both linear and quadratic effects (P < 0.05) of buffer pH on CMCase activity were significant, with CMCase activity being maximal at pH 5.4-6.1. CMCase activity was higher (P < 0.05) at a CaCl2 concentration of 1 mmol L(-1) compared with lower values. CONCLUSION: Although sonication treatment increases the amount of protein extracted from rumen bacteria adhered to the residue of incubation, the CMCase activity of the released protein might be measured without sonication treatment and should be carried out with a hypotonic buffer solution that includes a calcium source. When pH is not a treatment factor, the buffer pH should be between 5.5 and 6.
Assuntos
Bactérias/metabolismo , Proteínas de Bactérias/metabolismo , Cloreto de Cálcio/farmacologia , Celulase/metabolismo , Digestão , Rúmen/microbiologia , Sonicação/métodos , Animais , Bactérias/enzimologia , Soluções Tampão , Cálcio da Dieta/farmacologia , Bovinos , Digestão/efeitos dos fármacos , Fermentação , Glicosídeo Hidrolases/metabolismo , Concentração de Íons de Hidrogênio , Masculino , Rúmen/metabolismoRESUMO
Sepsis is a potentially deadly complication that can be caused by different factors. Actually, it is known that oxidative stress is involved in the pathogenesis of sepsis. Thus, the aim of this study was to evaluate the effect of diphenyl diselenide (PhSe)(2), an emergent compound, on oxidative stress parameters induced by sepsis in rats. Animals were pre-injected with (PhSe)(2) or vehicle. Twenty-four hours later, sepsis was induced by cecal ligation puncture (CLP). After 12 h, liver was taken for thiobarbituric acid reactive species (TBARS) measurement, δ-aminolevunic acid dehydratase (δ-ALA-D), Cu/Zn superoxide dismutase (Cu/Zn SOD) and catalase (CAT) activities assay. The sepsis increased TBARS, inhibited δ-ALA-D, activated Cu/Zn SOD and had a tendency to decrease CAT activity. However, (PhSe)(2) prevented the TBARS formation, but did not prevent the inhibition of δ-ALA-D activity in the animals with damage. Thus, this study showed that (PhSe)(2) partially prevents the oxidative stress induced by sepsis, indicating the potential of this compound as a treatment for this pathology. Nevertheless, more tests should be performed to confirm the hypothesis suggested here.