Lactate dehydrogenase isoenzyme distribution and patterns in chicken organs.

Unlike most mammals, chicken lactate dehydrogenase isoenzymes cannot be separated using the 'Titan-Gel' electrophoresis. However, using isoelectric focusing at a pH range of 3.0 to 9.0, a good and clear separation of all five isoenzymes was achieved. Generally, three characteristic groups were seen: (a) those having a cathodic domination (breast muscle and serum) with mainly lactate dehydrogenase-5 (b) those having an anodic domination (heart, muscle, liver, pancreas, kidney, erythrocytes) of mainly lactate dehydrogenase - 1 and 2 and (c) those with a more uniform distribution (spleen, lung, and brain). The total lactate dehydrogenase activity was the highest in the breast muscle, followed by the heart muscle, liver and serum with the lowest activities in the lung and pancreas.

Antioxidant and detoxifying enzymes in the liver and kidney of pheasants after intoxication by herbicides MCPA and ANITEN I.

The activity of antioxidant and detoxifying enzymes, such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSHPx), glutathione reductase, glutathione-S-transferase (GST), the contents of thiobarbituric acid reactive substances, and the superoxide dismutase and glutathione-S-transferase isoenzyme patterns, were determined in the liver and kidney of pheasants after acute intoxication by herbicides MCPA and ANITEN I. In the liver, the activity of antioxidant enzymes was significantly decreased in the group given ANITEN I. New superoxide dismutase isoforms (pI 6.30, 6.85, 7.00) and higher intensity of isoform with pI 6.60 were observed after isoelectrofocusing in all experimental groups. In the kidney, the activity of superoxide dismutase was significantly decreased, and a higher intensity of superoxide dismutase isoforms (pI 6.00 and 6.60) was observed in all experimental groups. The contents of thiobarbituric acid reactive substances were significantly increased in the group with ANITEN I. The glutathione-S-transferase isoenzyme pattern was studied by using subunit-specific substrates and by Western blotting. The activity of glutathione-S-transferase with ethacrynic acid and cross-reactivity with rat subunit 7 was lower in all experimental groups in the kidney and liver, except in the liver of the group given a higher dose of ANITEN I. In this group, we have found a 2.10-fold higher activity to ethacrynic acid and a strong induction of subunit 7.

Antioxidant and detoxifying fish enzymes as biomarkers of river pollution.

The activity of several antioxidant and detoxifying enzymes, superoxide dismutase SOD, GSH peroxidase GSHPx, GSSG reductase GSR and GSH S transferase GST, the contents of thiobarbituric acid reactive substances TBARS, and the SOD and GST isoenzyme patterns were studied in the livers of chubs Leuciscus cephalus from reference river areas and polluted urban sites. Livers of polluted fish contained higher concentrations of transition metals, especially copper and iron. Total GSHPx activity was 1.8 fold higher in the polluted fish than in reference animals, while the SOD and GSR activities and the TBARS content were not significantly changed. Three new SOD isoforms pI 4.45, 5.1, 5.2 and a higher intensity of the band pI 4.2 were observed after isoelectrofocusing of polluted fish extracts. Total GST activity was higher in fish from polluted areas. The GST isoenzyme pattern was studied using subunit specific substrates DCNB, EPNP, EA, NPB, NBC and by Western blot using antibodies specific to rat GST subunits 1, 8 Alpha class, 3 Mu class and 7 Pi class. Reference and polluted fish lacked cross reactivity towards Alpha class GSTs. Reference fish displayed weaker cross reactivity towards CST 7 and 2.3 fold lower activity with EA, while higher cross reaction with GST 3 was observed in polluted fish.

Changes in GST-isoenzyme pattern of some organs of sheep exposed to different levels of pollution.

GST isonzyme patterns were studied in the cytosolic fraction of liver, kidney and lung of sheep exposed to industrial metal pollutants and compared with those of control animals. The methodology included the determination of enzymatic activities with several subunit-specific substrates (DCNB, NPB, EPNP and EA) and Western blotting using antibodies to specific rat GST subunits 1, 8 (alpha class), 3 (mu class) and 7 (pi class). In liver and lung, crossed reactivities with subunits 1 and 3 were absent in the controls but were present in exposed animals. Just the opposite result was obtained for subunit 8 crossed reactivity that was only in the control animals. In the kidney, crossed reactivities towards subunits 3 and 8 were absent and crossed reactivity equivalent to subunit 7 was present in all animals, and equivalent to subunit 1 was weakly induced in exposed animals. A 3.3-fold increase in the activity with NPB detected in the kidneys of exposed animals points to the induction of a theta class isoenzymes. Clear increases were found in the livers of exposed animals in the activities with CDNB (1.8-fold), DCNB (2.6-fold) and EPNP (2.1-fold), but no differences were found in the lungs with any of the substrates. The GST isoenzyme pattern of liver and lung could be, in principle, a useful biomarker of exposure to environmental pollution in sheep.

Effects of iron and vitamin E administration on some immunological parameters in pigs.

The effects of intramuscular administration of iron and vitamin E on phagocytic functions of leucocytes and on T-lymphocyte percentages were studied. Twelve pigs with an average body weight (BW) of 40 kg were divided into four treatment groups: +E (20 mg tocopheryl acetate per kg BW); +Fe (10 mg iron dextran per kg BW); +E/+Fe (20 mg tocopheryl acetate + 10 mg iron dextran per kg BW), C (control). Blood samples were collected from the sinus ophthalmicus prior to administration the preparations, and then at 12, 24, 48, 72 hours, and 7 days after administration. The iron injection led to a significant increase (p < 0,05) in plasma iron concentrations in both groups given Ferridextran. An evaluation of the immunological parameters showed the greatest rise in T-lymphocyte percentages as well as in metabolic activity of phagocytes (tetrazolium reduction) in vitamin E treated groups (+E, +E/+Fe) within 24 hours after injection. No interactive effects between vitamin E and iron on the immunological parameters studied were found.

Indirect examination of exocrine pancreatic function in vivo by determination of chymotrypsin activity in animal intestine using a synthetic substrate.

Using the substrate N-acetyl-L-tyrosyl-p-aminobenzoic acid, we determined chymotrypsin activity in the small intestine of calf, pig, and poultry. Orally administered N-acetyl-L-tyrosyl-p-aminobenzoic acid is enzymatically cleaved in vivo, and the released p-aminobenzoic acid is determined by HPLC. We found that the p-aminobenzoic acid concentration in plasma and urine was significantly influenced by the feeding of soya flour. After soybean flour feeding, the p-aminobenzoic acid concentration significantly increased in the plasma of calves and hens, in contrast to pigs, where the p-aminobenzoic acid concentration significantly decreased. This shows that the oral administration of N-acetyl-L-tyrosyl-p-aminobenzoic acid with subsequent determination of p-aminobenzoic acid is suitable for the estimation of exocrine pancreatic function and for determination of changes in intestinal proteolytic activity caused by antinutritive substances.

Lactate dehydrogenase isoenzyme patterns in bird, carp and mammalian sera.

Isoenzyme patterns of lactate dehydrogenase (EC 1.1.1.27) in avian sera are described and compared with those of carp and some mammals. The predominant portion of lactate dehydrogenase in avian sera was concentrated in the muscular form of the enzyme (lactate dehydrogenase 5). Most mammalian sera (with the exception of rat serum) showed a different pattern, in which the main portion of lactate dehydrogenase activity migrated in the first three anodic fractions. Fish serum lactate dehydrogenase isoenzymes were distributed similarly to those of mammals. The electrophoretic mobility of bird lactate dehydrogenase isoenzymes in a pH gradient of 3 to 9 was different from that of carp, cattle and rabbit. Bird lactate dehydrogenase isoenzymes were localized in the pH region of 5.8 to 8.1. In contrast, the mammalian isoenzymes were more acidic, with pI values in the range of 4.5 to 7.3. Lactate dehydrogenase isoenzymes of carp migrated in a narrow pH range of 5.2 to 6.5.

Plasma transport and tissue distribution of [14C] beta-carotene and [3H]retinol administered orally to pigs.

The absorption, plasma transport, and tissue distribution of beta-carotene and vitamin A were studied in pigs after the oral administration of radiolabeled beta-carotene and retinol. Plasma radioactivity as well as retinol and beta-carotene were measured over a 24-h period, after which the pigs were killed. Plasma radioactivity increased within 4 h and 5 h for beta-carotene and retinol, respectively. The administration of radiolabeled retinol resulted in highest levels in the liver (83016 +/- 17614 dpm/g of tissue), followed by the kidney (5598 +/- 1309 dpm/g), the duodenum (2783 +/- 300 dpm/g), the colon (835 +/- 117 dpm/g), and the lung (756 +/- 68 dpm/g). Twenty-four hours after beta-carotene administration, tissue radioactivity distribution showed highest accumulation in the lung (22772 +/- 11997 dpm/g) and the liver (2328 +/- 694 dpm/g). The identification of the radioactive component 14C in the lung and colon indicated that the radioactivity was associated with beta-carotene only. This indicates that pigs may absorb intact beta-carotene. In other tissues such as liver, kidney, and intestine, only retinol-associated radioactivity was found after saponification of retinyl esters. The physiological importance of the unique accumulation of beta-carotene in lung tissue remains speculative.

Simultaneous determination of p-aminobenzoic acid and its metabolites in urine by high performance liquid chromatography.

We developed a new HPLC method for the determination of p-aminobenzoic acid (PABA) and its metabolites (p-aminohippuric acid, N-acetyl-p-aminohippuric acid, N-acetyl-p-aminobenzoic acid) in urine. As the internal standard m-hydroxybenzoic acid was used. In the isocratic elution the mobile phase consisted of methanol and 0.02 M ammonium acetate (20:80 v/v, pH 4.0). The separation was carried out on the C18, reversed-phase column, particle size 5 microns. The separated components were detected at 280 nm. The method can be used in the assessment of the response of pancrease (secretion of digestive enzymes) to soya feeding as well as in the diagnosis of the exocrine pancreatic diseases of animals.

[Dynamics of specific antibody formation in sheep after administration of industrial substrates from an aluminum plant]. / Dynamika tvorby specifických protilátok u oviec pocas príjmu priemyselného substrátu z hlinikárne.

Our observations aimed at determining the effects of supplementation with aluminium of plant emissions on specific ovalbumin antibody production in sheep by means of an ELISA method. Eleven Merino ewes aged 2.5 years were included in the experiment. The experimental group consisted of 6 animals. The daily intake of 0.75 g substrate per animal was administered after the morning feeding via a laryngeal tube. The amounts of essential and risk elements included in the substrate are given in Tab. I. All animals were subcutaneously immunized with ovalbumin (OVA, SIGMA A 5503) in 10% alhydrogel (Superhpos Ltd., Denmark) at a dose of 0.2 mg per 10 kg of live weight. The first immunization took place prior to the first gavage of emissions, the second one on day 21 of the experiment. Blood samples from the v. jugularis were collected from all animals, prior to the first immunization, in 6 weekly intervals and then in the 8th and 10th week of the experiment. A modified ELISA method (Strobel, 1983) was used to determine specific OVA antibodies in the sera. Throughout the observation period the increase of OVA antibody production appeared to be more significant in the experimental sheep. In the latter, increased specific antibody production could be detected as early as in the 1st week with maximum immunoglobulinaemia occurring in weeks 3 and 6 after OVA administration. As to specific antibody concentrations, significant differences between the experimental and the control ewes were recorded in weeks 2, 3, 4, 5 and 6 of the experiment (p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

[Levels of thyroxine, beta carotene and vitamin A during successful and unsuccessful insemination of heifers]. / Koncentracie tyroxínu, beta karoténu a vitamínu A pri úspesnej a neúspesnej inseminácii jaloviíc.

We studied the concentrations of thyroxine (T4), beta-carotene and vitamin A (vit. A) on the day (zero-th day) of the first successful and unsuccessful insemination of 63 heifers that were administered feed rations balanced according to the Czechoslovak standard CSN 46,7070, housed all the year round, inseminated at uniform time (8:00-9:00 a. m.) and with blood samplings at uniform time (10:00-12:00 a. m.). In the repeat-breeder heifers during the first insemination the above parameters were determined in the same conditions on the day of their second, or third successful and also unsuccessful insemination; the changes relevant to the first insemination were evaluated. In relation to the day of the first successful insemination, we recorded a significant increase in T4 concentration (P less than 0.001; P less than 0.001) on the day of the second, or third successful inseminations. During the second insemination we recorded significantly lower T4 concentrations in the cows which had failed to be inseminated in comparison with successfully inseminated cows; these concentrations did not differ significantly from T4 concentrations in the heifers that failed to become pregnant in the first insemination. Parallelly with the rise in T4 concentration, beta-carotene concentrations also increased during the second and third inseminations (P greater than 0.05). No difference was recorded in beta-carotene concentrations after the first and second unsuccessful inseminations.(ABSTRACT TRUNCATED AT 250 WORDS)

[Concentrations of thyroxine, beta carotene, vitamin A and pregnancy in heifers after administration of beta carotene]. / Koncentrácie tyroxínu, beta karoténu, vitamínu A a zabrezávanie jalovíc po prívode beta karoténu.

Serum concentrations of thyroxine (T4), beta-carotene (beta K), vitamin A and conception after the first insemination on the zero, sixth and 21st day were studied in 63 heifers fed synthetic beta K supplement in groups I to IV at stanchion housing and low-carotene diet. The following amount of beta K was supplemented per head/day: I--0 mg (control, n = 17), II--100 mg (n = 16), III--200 mg (n = 14) and IV--400 mg (n = 16). The parameters were evaluated separately according to groups and conception. By comparing the T4 concentration between experimental groups and control, separately in non-gravid and gravid animals, a significant increase (P less than 0.05) was determined only on the zero day in the fourth group. At the same time, in the fourth group, in comparison with the others, insignificantly lower T4 values were recorded in non-gravid animals. beta K values on the zero day in gravid animals increased in a direct proportion to the higher supplement (P less than 0.05; P less than 0.01; P less than 0.001); in non-gravid animals the increase was slower (P less than 0.05; P less than 0.05; P less than 0.01), but differences in relation to gravidity were not significant. An increase of beta K had no significant effect on serum concentrations of vitamin A. Differences between groups as far as conception was concerned were insignificant. It follows from the results that increasing rates of beta K, with the exception of serum concentrations, had no significant effect on other parameters. Higher concentrations of T4 and a slower increase of beta K in non-gravid animals, however, point at mutual co-operation relation and its positive effect on conception in animals.

[Stimulatory effect of vitamin A on T and B lymphocytes in experimental ascariasis in pigs]. / Stimulacný vplyv vitamínu A na T a B lymfocyty pri experimentálnej askaridóze prasiat.

The nonspecific, stimulative effect of vitamin A, applied per os, was studied on the T and B cell population in the course of post-invasive, migratory phase of experimental ascariasis of swine. The vitamin under study was found to exert an activating influence on the level of both immunocompetent cells already in the early phase of invasive process, with the maximum on the 9th, and/or 12th day after invasion. The protective effect of vitamin A, i. e. the effect on the reduction of migrating larvae of ascarids in the lungs, was 50%. This phenomenon is in accord with the present knowledge on the role of this vitamin in immunity.

Glutamate dehydrogenase and glutamine synthetase activity in some organs of ruminants and monogastric animals.

A comparative study of glutamate dehydrogenase (GLDH 1.4.1.2) and glutamine synthetase (GS 6.3.1.2.) activity in liver, kidney and spleen homogenates from cattle, sheep, pigs and chickens showed that chicken liver contained on an average 3.5%, pig liver 8.3% and bovine liver 45.6% of the glutamate dehydrogenase activity present in sheep liver. Relatively low trace activity was found in the spleen and kidneys, except for the renal cortex of cattle (32% of activity in the liver). GS activity was the highest in chicken liver; in pigs it amounted to 33.40%, in cattle to 24.2% and in sheep to 19.7% of this activity. No marked interspecies differences were found in the values in the kidneys and spleen. It can be concluded from the results that the relatively high GLDH activity in the liver of ruminants compared with pigs and chicken is associated with the greater ability of ruminants to utilize ammonia. The higher GS activity and lower GLDH activity in chicken liver can be attributed to higher uric acid synthesis from ammonia via glutamine and purine bases and the lower ability of birds to utilize ammonia for protein synthesis. The presence of alanine dehydrogenase was not demonstrated in chicken liver, where the maximum oxidation of NADH after the addition to pyruvate and ammonia substrate was found.

[Effect of a urea diet on glutamate dehydrogenase and glutamine synthetase activity in various organs of chickens]. / Vplyv mocovinovej diety na aktivitu glutamátdehydrogenázy a glutaminsyntetazy v niektorých orgánoch brojlerov

The total values were determined for the activity of glutamate dehydrogenase, glutamine synthetase, and dehydrogenase with pyruvate in broilers fed a diet with a 0, 2 and 4% content of urea for three weeks. A statistically significant increase of glutamate dehydrogenase activity was ascertained in the liver and kidney of broilers. The increase of the activity of glutamine synthetase in liver was close to the threshold of statistical significance. Dehydrogenase activity with pyruvate increased in liver.

[Year-round buffering of cattle feed mixture and its effect on metabolism and productivity]. / Celorocná pufrácia krmnej dávky výkrmového hovädzieho dobytka bentonitom a jej vplyv na metabolizmus a úzitkovost

Commercial feed mixture was buffered with a 2% and 3% admixture of bentonite buffer in two beef cattle herds in the course of one year. The mixtures were fed on a continuous basis. The two-per-cent buffer concentration was tested in 110 test animals with 104 control animals and the three-per-cent concentration in 50 test animals with 50 controls. Throughout the trial the over-all health condition remained unchanged, the hematocrit and hemoglobin values were balanced in both groups. The biochemical indices were better in the test groups: hypocalcemia improved (in the controls it grew worse), magnesiemia was slightly increased, the inorganic serum factor did not go beyond physiological limits, and acidosis did not occur (as distinct from the control animals). The levels of transaminases (GOT, GPT), glutamic acid dehydrogenase, total serum protein, alkaline phosphatase as well as ammonia and urea in blood serum were at physiological values with po-differences within groups. In the case of the three-per-cent buffer concentration the daily gains were higher by 0.073 kg, and in the two-per-cent concentration by 0.058 kg, in the test animals. The average annual gain was higher by 25.5 kg, and by 18.3 kg, respectively. With respect to the price of buffer and to the efficiency of the animals tested, the economic indices of feed mixture buffering are highly effective.