Comparative study of antibacterial activity of pancreatic juice in six mammalian species.

A comparative study of antibacterial activity of pancreatic juice was conducted on six mammalian species. Pancreatic juice collections were conducted as acute (rabbit, guinea pig, rat) and chronic (pig, sheep, cattle) experiments, in the former before and after stimulation [cholecystokinin (CCK) and secretin] and in the latter under basal conditions alone. Antibacterial activity was tested on Micrococcus pyogenes and compared with that of neomycin. The samples were tested under normal conditions and after heating and dilution. The pancreatic juice of rat showed no activity against Micrococcus pyogenes. The antibacterial activity of rabbit and guinea pig pancreatic juice under basal conditions was similar within the group but significantly higher than that of pig, sheep and cattle which also did not differ significantly within the group. On stimulation with CCK and secretin, no significant change could be observed in the potency of antimicrobial activity of pancreatic juice in the rabbit and guinea pig. The antibacterial activity remained unchanged after heating to 65 degrees C and upon dilution to 1:10.

The influence of nitrite, nitrate and nitric oxide on ammonia use and urea production in primary cultures of sheep hepatocytes.

Inorganic nitrite introduced into the living organism is rapidly converted into nitrate and nitric oxide (NO). It is known that nitrite decreases ammonia use and urea formation in vitro and it seems that nitrite also influences these processes in vivo. However, the mechanism underlying this effect is not known. Therefore, we decided to compare the influence of sodium nitrite (NaNO(2)), sodium nitrate (NaNO(3)) and NO on ammonia use and ureagenesis in monolayer cultures of sheep hepatocytes during 18 hr of cultivation. It was found that 0.5 and 2.5 mmNaNO(2) significantly reduced ammonia use in a dose-dependent manner for the first 6 hr of incubation; at higher concentrations (2.5 mm), it also decreased urea formation. Also, the presence of nitrite did not affect the lactate dehydrogenase (LDH) activity in the medium which indicates that the nitrite effect did not result from its cytotoxic action. NaNO(3) (0.5 and 2.5 mm) did not induce any changes in ammonia use and urea synthesis in hepatocytes. With sodium nitroprusside (SNP) (0.001, 0.01, 0.1, 0.5 and 1.0 mm) a decrease of ammonia use and urea production was observed corresponding to the nitrite effect, but contrary to nitrite exposure these changes in metabolism were persistent during the whole cultivation period. On the other hand, potassium cyanide (KCN) (0.1 and 0.5 mm) did not influence either urea formation or ammonia use. Thus, it can be concluded that in isolated hepatocytes nitrate and/or NO are not the mediators of nitrite effects on nitrogen metabolism. Furthermore, there is no evidence that nitrite effects are mediated by impaired mitochondrial respiration and energy production.

The influence of phentolamine on the hyperglycaemic and lipolytic effects of ammonia in sheep.

Intravenous infusion over 30 min of ammonium chloride [10-1 g/kg b.w.] caused a significant increase of blood glucose and free fatty acids levels, and a similar effect was observed with adrenaline. Phentolamine [1 mg/kg b.w.] abolished the hyperglycaemic action of ammonium chloride but only slightly diminished the lipolytic effect. It is concluded that the ammonium ion influences carbohydrate metabolism chiefly by way of catecholamines and stimulation of alpha-adrenergic receptors, while tissue lipids are mobilized by ammonia by another route, possibley directly.

Comparison of metabolic effect of ammonia and adrenaline infusions in sheep.

Intravenous infusions of ammonium chloride (62.3 mumol.kg-1.min-1) for 30 min caused a significant increase in blood glucose, lactate, pyruvate and free fatty acid (FFA) levels. A similar effect was also observed during infusion of adrenaline. Propanolol--a beta-receptor blocking agent--completely prevented the rise of blood pyruvate and lactate after adrenaline when 8.3 microgram.kg-1.min-1 of propranolol were infused, but not after NH4Cl administration. Lipolytic actions of adrenaline were completely prevented but that of NH4Cl was only significantly diminished by blockade of beta-receptors with propranolol. It was concluded that the influence of ammonium ions on blood lactate and pyruvate and FFA was not entirely mediated by adrenaline.

[Effects of carbachol and atropine on bile secretion in sheep]. / Wplyw karbacholu i atropiny na produkcje zólci u owiec.

Experiments were performed on rams prepared surgically to make possible the control of bile flow and the maintenance of enterohepatic circulation of bile components. Carbachol and atropine were infused into the jugular vein for 2 hours in the amounts: 0.3 and 0.7 microgram/kg b.w./min. respectively. Carbachol administration did not affect significantly the bile production. Atropine led to the significant decrease in the bile flow (p less than 0.01) by diminished concentration of bile acids in the bile. On the basis of the obtained results the role of n. vagus in the regulation of biliary secretion is discussed. It is suggested, that in physiological conditions the biliary secretion is, to some extent, maintained as a result of the muscarinic receptor stimulation. This stimulation attains the maximum level and it is not possible to evoke rise in bile production after either n. vagus stimulation or administration of parasympathomimetic agents.

Utilization of nitrogen from 15NH4Cl and [15N]alanine for urea synthesis in hepatocytes from fed and starved rats.

Utilization of N from 15NH4Cl and [15N]alanine for urea synthesis in hepatocytes isolated from fed and 24 hr starved rats was investigated. In hepatocytes isolated from fed rats, 54 and 65% of the added [15N]ammonia was utilized for urea synthesis in the presence of 0.5 and 2.0 mM NH4Cl, respectively. This utilization of [15N]ammonia in hepatocytes from starved rats was 2-fold lower. The amount of urea synthetized from endogenous sources was, in the presence of 0.5 and 2.0 mM NH4Cl, about 44 and 60% higher than in the control conditions (without NH4Cl). The considerable amount of added ammonia (30-44%) was utilized in processes other than urea synthesis. Alanine markedly diminished the utilization of 15N from NH4Cl in hepatocytes from both fed and starved rats. In these conditions (NH4Cl present), alanine significantly increased the urea formation in hepatocytes from starved rats and failed to affect the urea production in hepatocytes from fed rats. On the basis of 15N determination, it was concluded that both NH4Cl and alanine caused an increase in the utilization of nitrogen from endogenous sources in rat hepatocytes. This conclusion is in contrast with the results based only on the changes in ammonia and urea concentrations.

The changes in bile flow and its composition in sheep following liver damage by carbon tetrachloride.

Sheep were prepared surgically in a manner permitting the measurement of bile flow by preserving its enterohepatic circulation. Carbon tetrachloride (CCl4) was administered intraruminally in a dose 0.5 ml/kg b.w. After CCl4 administration a decrease in bile flow and bile acid excretion was observed. The decrease in bile flow observed after CCl4 administration was lower than the decrease in bile acid output. Thus it can be suggested that in this condition the effects of bile acid output on the bile secretion can be partially abolished and maintaining of relatively high bile flow is connected with an increase in the bile acid-independent bile flow. The increase of the bilirubin concentration in bile was observed after acute liver damage by carbon tetrachloride.

Differential behavioural and hormonal responses to two different stressors (footshocking and immobilization) in sheep.

Two different stressors footshocking and immobilization applied for 3 days induced in sheep differential behavioural and hormonal responses in cortisol and prolactin secretion. Immobilization drastically disturbed rumen motoric activity (significantly attenuated its contractions), caused loss of appetite and severe general depression. Footshocking did not induce any of these symptoms. Immobilization induced 2-3 fold higher rise of plasma cortisol mesors on the days of stressing as compared to footshocking. On the poststressing days after footshocking as well as after immobilization plasma cortisol mesors fell to 50% of the prestimulatory values. The rise of plasma prolactin mesors in footshocked and in immobilized animals on the days of stressing was nearly of the same magnitude. However, a significant difference in the response of prolactin secretion between footshocked and immobilized animals occurred in the circadian rhythmicity of the hormone secretion. Footshocking induced circadian rhythmicity with characteristic acrophases, while immobilization did not induce the circadian pattern of prolactin secretion. An attempt of interpretation of the described phenomena has been undertaken.

The fate of nitrogen from 15N-labeled nitrate after single intravenous administration of Na15NO3 in sheep.

The metabolic fate of nitrogen from 15N-labeled sodium nitrate has been investigated in four healthy Polish Merino ewes. 15N-labeled sodium nitrate was administered intravenously at the dosage of 400 micromol.kg(-1) body weight. Blood plasma and urine concentrations of nitrate, ammonia, and urea and 15N enrichment of ammonia and urea were estimated over a 50-h period following 15N-nitrate administration. Nitrate (NO3-) was slowly eliminated from the blood plasma, and the presence of NO3(-) in the blood plasma above the nitrate "background" was observed for 50 h. 15N enrichment of blood plasma urea already appeared at 15 min and reached the maximum 6 h after 15N-nitrate administration. The urinary excretion of nitrate occured during 50 h after 15N-nitrate injection; the total urine excretion of NO3(-) was 23.63+/-2.39% of the administered dose. The mean urinary recoveries of nitrogen as 15N-urea and 15N-ammonia were 14.76+/-1.32% and 0.096+/-0.015% of the administered 15N-nitrate dose, respectively. It should be pointed out that in total only 38.49% of the administered nitrate-N was excreted in urine (as nitrate, ammonia and urea nitrogen) during 50 h. The results obtained indicate that sheep are able to store nitrate nitrogen in their body. The fate of the remaining approximately 60% of the 15NO3(-) administered dose is unknown. The results obtained do not allow one to conclude what fraction of the unrecovered approximately 60% of the 15NO3(-) dose was utilized by gastrointestinal microorganisms, and (or) metabolized, or stored in sheep tissues.

Effect of nitrite on ureagenesis and carbohydrate metabolism in isolated rat hepatocytes.

The effect of three NaNO2 concentrations (0.5, 2.0, and 5.0 mM) on 15N-ammonia utilization, ureagenesis, glucose, pyruvate and lactate formation and glycogen breakdown were studied in isolated rat hepatocytes. Nitrite failed to affect the rate of glycogenolysis as well as the lactate and pyruvate formation, but at the same time it markedly increased the glucose formation. It is concluded that the increase in the glucose formation results from the nitrite stimulation of the rate of gluconeogenesis. An increased sodium nitrite concentration caused a significant decrease in the ammonia utilization and urea synthesis; there are strong linear correlations between the nitrite concentration and the amount of utilized ammonia (r = -0.93) and the formed urea (r = -0.96). The observed lower rate of ureagenesis in the presence of nitrite resulted from the diminished incorporation of the added 15N-ammonia into urea, as well as from the diminished urea formation from endogenous nitrogen. It is concluded that the disturbances in carbohydrate and nitrogen metabolism observed in the nitrite-poisoned animals are attributed to the direct effect of nitrite on metabolism.

Influence of feeding regimen and postnatal developmental stages on antibacterial activity of pancreatic juice.

Antibacterial activity of pancreatic juice in the pig (n = 8) was investigated during early postnatal development and in cattle (n = 6) receiving a different feeding regimen. For pancreatic juice collection, a catheter was surgically implanted in the pancreatic duct. Reintroduction of pancreatic juice was achieved through a T-shaped cannula in the duodenum. Pancreatic juice was collected for 30 min in all cases. In piglets, collections were carried out at 2, 5-6, and 7-10 wk of age, and in cattle, after a standard meal, 48 h starvation, and following 24 h intraduodenal glucose infusion. Antibacterial activity was tested on Micrococcus Pyogenes strain ATTC 6538P by disc agar diffusion technique using nonactivated pancreatic juice, before and after heat treatment for 15 min at 65 and 100 degrees C, respectively. Piglets showed a significant rise in antibacterial activity of pancreatic juice after weaning. In cattle, 48 h of starvation resulted in a marked suppression of antibacterial activity. This activity was found to be normal after a standard meal and comparable to that after 24-h intraduodenal glucose infusion. Heating of pancreatic juice to 65 degrees C caused a 35% increase in the antibacterial potency, whereas heating to 100 degrees C completely abolished it. Additionally, dilution of pancreatic juice to 1:10 did not affect antibacterial potency.

In vitro ammonia utilization and urea synthesis by rat liver after portacaval shunt.

BACKGROUND: The objective of this study was to study in vitro ammonia utilization and urea synthesis by the rat liver after portacaval shunt (PCS) in comparison to controls. This study is part of a series of experiments to investigate metabolic derangements in the liver following PCS. METHODS: Sixteen male Wistar rats weighing 150-230 g were used. PCS was performed on 8 and the rest served as sham-operated controls. The experiments were performed 4 weeks postoperatively. The usually monitored postoperative parameters after PCS like body weight, rise in ammonia levels in the peripheral blood, atrophy of mesenteric fat, atrophy of the liver, etc., were comparable to other PCS series. Liver slices (1 x 2 x 1 mm) were prepared from each rat liver and incubated in Krebs-Henseleit buffer. RESULTS: After incubation, ammonia utilization in the controls was 162.88+/-20.12 micromol/g dry weight/h while in PCS rats it was 81.35+/-2.64 micromol/g dry weight/h (p < 0.0001). The rate of urea synthesis in controls was 122.54+/-12.93 micromol/g dry weight/h whereas in the PCS group it was 14.99+/-2.21 micromol/g dry weight/h (p < 0.0001). CONCLUSION: These results indicate a significant decline in the capacity of the liver for ammonia utilization as well as urea synthesis after PCS.

A study of the pharmacokinetics and tissue residues of an oral trimethoprim/sulphadiazine formulation in healthy pigs.

Twenty-six healthy female pigs weighing 19.5-33 kg were used in three separate experiments. The animals were fed individually twice a day. Trimethoprim/sulphadiazine (TMP/SDZ) formulation was added to feed in the amount of 6 mg/kg bw (TMP) and 30 mg/kg bw (SDZ). TMP and SDZ concentrations in blood plasma, muscles, liver and kidneys were measured. Pharmacokinetic parameters show that the absorption of TMP from the alimentary tract in pigs is faster than the absorption of SDZ, and the elimination of TMP is slower than that of SDZ. The absorption half-lives were 0.96 (TMP) and 2.24 h (SDZ), whereas elimination half-lives were 5.49 (TMP) and 4.19 h (SDZ). The observed TMP:SDZ ratios in blood plasma after multiple dose administration ranged from 1:11.4 to 1:23.2. One day after administration of the last dose of TMP/SDZ the plasma concentration ratio was 1:15.5, but in muscles, liver and kidneys it was much lower: 1:0.79, 1:0.14 and 1:1.53 respectively. The absolute TMP and SDZ tissue concentrations 1 day after the last multiple dose administration were very low (maximum TMP: 0.29 micrograms/g in liver; maximum SDZ: 0.23 micrograms/g in kidneys). Neither drug was detected in any tissue 8 days after the last administration of TMP/SDZ. Based on our results, it was concluded that there is no support for the TMP:SDZ pharmaceutical ratio 1:5 in oral formulations of these compounds for pigs. The administration oral TMP/SDZ formulations once a day may result in the absolute tissue concentrations of these drugs being too low for antibacterial activity. The withdrawal period for such an oral TMP/SDZ formulation for pigs (according to accepted guidelines in Europe for MRL of TMP < 0.05 mg/kg of tissue) should not be less than 5 days.

Effect of propionate on the utilization of nitrogen from 15NH4Cl for urea synthesis in hepatocytes isolated from sheep liver.

1. The effect of ornithine (2.0 mM) and propionate (5.0 mM) on the utilization of N from 15NH4Cl (5.0 mM) for urea synthesis in hepatocytes isolated from sheep liver was investigated. 2. The capacity of sheep hepatocytes to utilize [15N]ammonia in the absence of the other exogenous substrates was very low and amounted 132 +/- 37.3 mumol/hr per 1 g dry wt. 3. Ornithine failed to affect the total [15N]ammonia uptake and total urea synthesis, but at the same time it markedly increased the utilization of [15N]ammonia for ureagenesis and diminished the rate of urea synthesis from endogenous sources. 4. Propionate markedly increased total [15N]ammonia utilization and total urea formation; this increase resulted from the rise of ammonia utilization for urea synthesis and it was similar in the presence or absence of ornithine. 5. The capacity of sheep liver cells to utilize ammonia in the presence of propionate (in the presence or absence of ornithine) amounted to 256 mumol/hr per 1 g dry wt, thus being similar to the values in vivo. 6. It is concluded that in sheep hepatocytes both ornithine and propionate stimulate the utilization of ammonia for urea synthesis and these effects take place independently and occur by different mechanisms.