Efeitos da defaunaçäo em ovinos alimentados com cana-de-açúcar (Saccharum officinarum, L. ) adicionada de uréia / Effects of defaunation in sheep fed sugar cane (Saccharum officinarum, L. ) plus urea

Em delineamento inteiramente ao acaso, estudou-se o efeito da defaunaçäo em ovinos que receberam cana-de-açúcar e uréia sobre o consumo e digestibilidade aparente de nutrientes, dinâmica da fase sólida, parâmetros de fermentaçäo ruminal e de degradaçäo in situ. As taxas de passagem no rúmen foram, respectivamente, de 3,05 e 1,97 por cento/h, para ovinos faunados e defaunados. Valores mais elevados para taxa (5,4 por cento/h) e extensäo (46,7 por cento) da digestäo ruminal e de degradabilidade efetiva da FDN da cana (31,5 por cento) foram obtidos nos ovinos faunados. Seus consumos diários (57,7 e 32,3 g/kg0,75, respectivamente para MS e FDN) foram superiores (P<0,06) aos dos ovinos defaunados. Em funçäo do tempo de amostragem observaram-se maiores (P<0,05) concentrações de AGV total, acetato e propionato para os cordeiros faunados. Nos defaunados ocorreu queda pós-prandial mais acentuada (P<0,04) no pH. Para digestibilidade aparente diferença (P<0,05) entre tratamentos foi verificada apenas para matéria seca

Bradykinin, but not muscarinic, inhibition of M-current in rat sympathetic ganglion neurons involves phospholipase C-beta 4.

Rat superior cervical ganglion (SCG) neurons express low-threshold noninactivating M-type potassium channels (I(K(M))), which can be inhibited by activation of M(1) muscarinic receptors (M(1) mAChR) and bradykinin (BK) B(2) receptors. Inhibition by the M(1) mAChR agonist oxotremorine methiodide (Oxo-M) is mediated, at least in part, by the pertussis toxin-insensitive G-protein Galpha(q) (Caulfield et al., 1994; Haley et al., 1998a), whereas BK inhibition involves Galpha(q) and/or Galpha(11) (Jones et al., 1995). Galpha(q) and Galpha(11) can stimulate phospholipase C-beta (PLC-beta), raising the possibility that PLC is involved in I(K(M)) inhibition by Oxo-M and BK. RT-PCR and antibody staining confirmed the presence of PLC-beta1, -beta2, -beta3, and -beta4 in rat SCG. We have tested the role of two PLC isoforms (PLC-beta1 and PLC-beta4) using antisense-expression constructs. Antisense constructs, consisting of the cytomegalovirus promoter driving antisense cRNA corresponding to the 3'-untranslated regions of PLC-beta1 and PLC-beta4, were injected into the nucleus of dissociated SCG neurons. Injected cells showed reduced antibody staining for the relevant PLC-beta isoform when compared to uninjected cells 48 hr later. BK inhibition of I(K(M)) was significantly reduced 48 hr after injection of the PLC-beta4, but not the PLC-beta1, antisense-encoding plasmid. Neither PLC-beta antisense altered M(1) mAChR inhibition by Oxo-M. These data support the conclusion of Cruzblanca et al. (1998) that BK, but not M(1) mAChR, inhibition of I(K(M)) involves PLC and extends this finding by indicating that PLC-beta4 is involved.

G-proteins and G-protein subunits mediating cholinergic inhibition of N-type calcium currents in sympathetic neurons.

One postsynaptic action of the transmitter acetylcholine in sympathetic ganglia is to inhibit somatic N-type Ca2+ currents: this reduces Ca2+-activated K+ currents and facilitates high-frequency spiking. Previous experiments on rat superior cervical ganglion neurons have revealed two distinct pathways for this inhibitory action: a rapid, voltage-dependent inhibition through activation of M4 muscarinic acetylcholine receptors (mAChRs), and a slower, voltage-independent inhibition via M1 mAChRs [Hille (1994) Trends in Neurosci., 17, 531-536]. We have analysed the mechanistic basis for this divergence at the level of the individual G-proteins and their alpha and betagamma subunits, using a combination of site-directed antibody injection, plasmid-driven antisense RNA expression, overexpression of selected constitutively active subunits, and antagonism of endogenously liberated betagamma subunits by over-expression of Dy-binding P-adrenergic receptor kinase 1 (PARK1) peptide. The results indicate that: (i) M4 mAChR-induced inhibition is mediated by GoA; (ii) a and Py subunits released from the activated GoA heterotrimer produce separate voltage-insensitive and voltage-sensitive components of inhibition, respectively; and (iii) voltage-insensitive M1 mAChR-induced inhibition is likely to be mediated by the alpha subunit of Gq. Hence, Ca2+ current inhibition results from the concerted, but independent actions of three different G-protein subunits.

The alpha subunit of Gq contributes to muscarinic inhibition of the M-type potassium current in sympathetic neurons.

Rat superior cervical ganglion (SCG) neurons express low-threshold noninactivating M-type potassium channels (IK(M)), which can be inhibited by activation of M1 muscarinic receptors. This inhibition occurs via pertussis toxin-insensitive G-proteins belonging to the Galphaq family (Caulfield et al., 1994 ). We have used DNA plasmids encoding antisense sequences against the 3' untranslated regions of Galpha subunits (antisense plasmids) to investigate the specific G-protein subunits involved in muscarinic inhibition of IK(M). These antisense plasmids specifically reduced levels of the target G-protein 48 hr after intranuclear injection. In cells depleted of Galphaq, muscarinic inhibition of IK(M) was attenuated compared both with uninjected neurons and with neurons injected with an inappropriate GalphaoA antisense plasmid. In contrast, depletion of Galpha11 protein did not alter IK(M) inhibition. To determine whether the alpha or beta gamma subunits of the G-protein mediated this inhibition, we have overexpressed the C terminus of beta adrenergic receptor kinase 1 (betaARK1), which binds free beta gamma subunits. betaARK1 did not reduce muscarinic inhibition of IK(M) at a concentration of plasmid that can reduce beta gamma-mediated inhibition of calcium current (). Also, expression of beta1gamma2 dimers did not alter the IK(M) density in SCG neurons. In contrast, IK(M) was virtually abolished in cells expressing GTPase-deficient, constitutively active forms of Galphaq and Galpha11. These data suggest that Galphaq is the principal mediator of muscarinic IK(M) inhibition in rat SCG neurons and that this more likely results from an effect of the alpha subunit than the beta gamma subunits of the Gq heterotrimer.

On the role of endogenous G-protein beta gamma subunits in N-type Ca2+ current inhibition by neurotransmitters in rat sympathetic neurones.

1. Using whole-cell and perforated-patch recordings, we have examined the part played by endogenous G-protein beta gamma subunits in neurotransmitter-mediated inhibition of N-type Ca2+ channel current (ICa) in dissociated rat superior cervical sympathetic neurones. 2. Expression of the C-terminus domain of beta-adrenergic receptor kinase 1 (beta ARK1), which contains the consensus motif (QXXER) for binding G beta gamma, reduced the fast (pertussis toxin (PTX)-sensitive) and voltage-dependent inhibition of ICa by noradrenaline and somatostatin, but not the slow (PTX-insensitive) and voltage-independent inhibition induced by angiotensin II. beta ARK1 peptide reduced GTP-gamma-S-induced voltage-dependent and PTX-sensitive inhibition of ICa but not GTP-gamma-S-mediated voltage-independent inhibition. 3. Overexpression of G beta 1 gamma 2, which mimicked the voltage-dependent inhibition by reducing ICa density and enhancing basal facilitation, occluded the voltage-dependent noradrenaline- and somatostatin-mediated inhibitions but not the inhibition mediated by angiotensin II. 4. Co-expression of the C-terminus of beta ARK1 with beta 1 and gamma 2 subunits prevented the effects of G beta gamma dimers on basal Ca2+ channel behaviour in a manner consistent with the sequestering of G beta gamma. 5. The expression of the C-terminus of beta ARK1 slowed down reinhibition kinetics of ICa following conditioning depolarizations and induced long-lasting facilitation by cumulatively sequestering beta gamma subunits. 6. Our findings identify endogenous G beta gamma as the mediator of the voltage-dependent, PTX-sensitive inhibition of ICa induced by both noradrenaline and somatostatin but not the voltage-independent. PTX-insensitive inhibition by angiotensin II. They also support the view that voltage-dependent inhibition results from a direct G beta gamma-Ca2+ channel interaction.

Duodenal flow of microbial and feed nitrogen in sheep fed normal soybean meal or soybean meal treated with modified zein.

The effect of protecting soybean meal from microbial degradation in the rumen on duodenal flow of microbial N and feed N was studied with sheep. The soybean meal was protected with chemically modified zein. Two groups of four wethers, each equipped with a ruminal cannula and a duodenal reentrant cannula, were fed a diet based on corn silage (1 kg of DM/d) that contained either a normal or protected soybean meal supplement. The results showed no appreciable differences between the two supplements in ruminal fluid pH, total N, and NAN concentrations; however, the concentration of total VFA was lower for protected soybean meal than for normal soybean meal. Although the flow of NAN into the duodenum was not affected, bacterial N flow was 18% lower, and feed N flow was 195% higher, for protected soybean meal than for normal soybean meal. The digestibilities of OM, ADF, and N in the digestive tract were not affected by the type of supplement. The treatment of soybean meal decreased the degradability of total feed N in the stomach by 22 percentage points, but the increased supply of feed N into the small intestine because of the treatment was at the expense of decreased bacterial synthesis of protein in the rumen, probably because of a shortage of RDP in the diet.

Toxidez de cobre em carneiros sem protozoários no rúmen / Copper toxicity in defaunated sheep

Quatro carneiros, sem protozoários no rúmen, receberam dieta com 17 ppm de cobre. Após quatro meses nesta alimentaçäo, um animal morreu e outro foi sacrificado. Aos cinco meses, os outros dois animais morreram. Foram feitas necropsias em dois animais e amostras de fígado foram obtidas para análise de cobre. Os achados mais importantes à necropsia foram: mucosa e pele ictérica; rim com coloraçäo metálica e aumentado de volume, e vesícula com suco biliar de coloraçäo verde. O nível médio de cobre encontrado no fígado foi de 3451 ppm. Tanto os achados de necropsia quanto o nível médio de cobre no fígado säo indicativos de animais apresentando intoxicaçäo por cobre

Effects of bentonite on wool growth and nitrogen metabolism in fauna-free and faunated sheep.

Two experiments were carried out with sheep that originated from a fauna-free flock and were fed a soybean meal-corn silage diet with or without a bentonite supplement. One-half of the sheep fed each diet in each experiment were faunated with a mixed population of ruminal protozoa, whereas the other half of the sheep remained fauna-free until the end of both experiments. Wool growth and daily gain were measured in Exp. 1. (eight rams per treatment), which lasted 110 d, and the metabolic effects in the rumen and intestinal tract of protozoa and dietary bentonite supplement were tested with cannulated wethers (four wethers per treatment) in Exp. 2. The results of Exp. 1 showed decreased wool growth (P less than .05) due to the presence of protozoa in the rumen. Dietary supplementation with bentonite partly offset the decreased wool growth in sheep with protozoa, but there were no effects of dietary bentonite and no protozoa x bentonite interaction (P greater than .05). Daily gain was decreased by the dietary bentonite (P less than .05) supplement but was not affected (P greater than .05) by the ruminal presence of protozoa. In Exp. 2, protozoa increased (P less than .01) the ruminal concentrations of ammonia and decreased (P less than .05) the acetic:propionic acid molar ratio. Fractionation of N in the duodenal digesta flowing from the stomach to the small intestine showed that protozoa decreased (P less than .05) the flow of nonammonia N and bacterial N, and there was a protozoa x bentonite interaction for these effects (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of bentonite and monensin on selected elements in the stomach and liver of fauna-free and faunated sheep.

Forty-eight rams, originating from a fauna-free flock, were divided into three groups of 16 and fed a corn silage-based diet that was unsupplemented (control) or included bentonite or monensin supplements. Eight rams in each group were inoculated with a mixed population of ruminal protozoa; the other rams remained free of protozoa throughout the 110-d experiment. The rams had free access to drinking water and assigned diet. All rams were killed at the end of the experiment, and ruminal and abomasal contents and livers were removed and sampled. Protozoal numbers in ruminal fluid of faunated rams were lower for groups fed bentonite or monensin supplements than for the control group. Bentonite decreased and monensin increased ruminal pH. The ruminal solubilities of Cu, Zn, and Mg were decreased by the presence of ruminal protozoa, but those of Fe, Mn, and Ca were not affected. Bentonite supplement decreased, and monensin supplement increased, the ruminal solubilities of Cu, Zn, and Mg. Protozoa increased the abomasal solubilities of Fe, but the other elements were not affected. Liver concentrations of Cu were decreased by bentonite and increased by monensin, but protozoa decreased the liver concentrations of both Cu and Mg. Liver concentration of Zn was affected by a monensin x protozoa interaction and that of Mg by a bentonite x protozoa interaction. It was concluded that chronic Cu poisoning could be accelerated by dietary supplements of monensin in sheep without ruminal microfauna, and the dietary Cu bioavailability could be decreased by dietary supplements of bentonite in sheep with a normal population of protozoa in the rumen.