Influence of various concentrations of phenol and its derivatives on the activity of fish intestinal peptidases

The effects of phenol and its derivatives (4-chlorophenol, 4-nirophenol and 2,4-dinitrophenol) on the activity of peptidases in five species of fish differing in feeding modes have been studied. The power of the effects depends on the fish species and localization of the enzyme (mucosa or chyme). As a rule, in the in vitro conditions the phenols in concentrations of 0.06?0.5 μM/L considerably decreases activities of the intestinal peptidases in bream; silver bream; and, especially, in pike. In some cases, phenol and its derivatives induce an inconsiderable increase in peptidase activities. In zander and perch, in fact, no changes in the enzymatic activity under impact of phenols are observed. It is suggested that the effect of phenol and its derivatives depends to a high extent on the species specificity of peptidases: in the fish of fam. Percidae, the enzymes are relatively tolerant; in fam. Cyprinidae and Esocidae, they are sensitive to the studied toxicants.     

Metallothionein levels in sperm of various fish species

In the present paper we determined low molecular heavy metal binding protein called metallothionein (MT) in fish spermatozoa sampled from four fish species ( Acipenser baerii, Acipenser ruthenus, Perca fluviatilis and Sander lucioperca ). The spermatozoa samples were analyzed with differential pulse voltammetry DPV (Brdicka reaction). The average content of MT in the fish sperm was 0.64 ± 0.55 μ m . The highest content of MT was determined in spermatozoa of S. lucioperca (0.96 ± 0.86 μ m ), followed by P. fluviatilis (0.88 ± 0.11 μ m ), A. ruthenus (0.57 ± 0.60 μ m ) and A. baerii (0.43 ± 0.25 μ m ). The differences in the content of MT was probably related to the differences in age and perhaps to the health status based on the environmental conditions in which the tested specimens developed while also other still unknown factors may have been involved.     

Localization and activity of various peptidases in germinating barley

Summary Germinating barley grains contain at least eight different peptidases: three carboxypeptidase (pH optima 4.8, 5.2, and 5.7), three aminopeptidases which act on aminoacyl--naphthylamides (pH opitima in the hydrolysis of di- and tripeptides at pH 5.8–6.5), and two peptidases which hydrolyse Ala-Gly and Leu-Tyr optimally at pH 7.8 and 8.6 respectively. We have determined the activities of these enzymes in the different tissues of non-germinated grains and followed the changes in the activities during a 5-day germination at 16°C.The aleurone layers contain high activities of all three groups of peptidases; there are no changes in the activities of the five aminopeptidases on germination, while the carboxypeptidases exhibit a small increase of activity. The starchy endosperms contain high carboxypeptidase activities, which increase during germination, but are totally devoid of the five aminopeptidases.All the peptidases exhibit high activities in the scutella; the carboxypeptidases and the enzymes acting on Ala-Gly and Leu-Tyr increase in activity during germination, while the naphthylamidase activities remain constant.The three peptidase groups occur in the seedling as well, but compared to the other tissues the carboxypeptidase activities are very small and the naphthylamidase activities are very high. The last-named enzymes seem to be characteristic for growing tissues.The starchy endosperm contains about two thirds of the total reserve proteins of the grain. Its internal pH during germination is 5.0–5.2, a value at which all the carboxypeptidases are highly active. As these enzymes are present in high concentrations in this tissue, it is probable that they have a central role in the mobilization of the reserve proteins during germination. The high peptidase activities of the scutellum, on the other hand, suggest that some of the hydrolysis products are absorbed as peptides and these are further hydrolysed to amino acids in this tissue.Abbreviations used DTT dithiothreitol - GA₃ gibberellic acid - -NA -naphthylamide - TNBS 2,4,6-trinitrobenzene sulphonic acid - Z- N-carbobenzoxy     

A comparative study on innate immune parameters in the epidermal mucus of various fish species

Fish epidermal mucus and its components provide the first line of defense against pathogens. Little is known about the role of epidermal mucus enzymes in the innate immune system of fish species such as Arctic char (Salvelinus alpinus), brook trout (S. fontinalis), koi carp(Cyprinus carpio), striped bass (Morone saxatilis), haddock, (Melanogrammus aeglefinus), Atlantic cod (Gadus morhua) and hagfish (Myxine glutinosa). The epidermal mucus samples from these fish were analysed for the specific activities of various hydrolytic enzymes including lysozyme, alkaline phosphatase, cathepsin B and proteases and the enzyme levels were compared among the fish species. Of all the species hagfish mucus showed a high activity for lysozyme and proteases and koi carp mucus had the highest levels of alkaline phosphatase and cathepsin B. A wide variation in enzyme activities was observed among the seven species and also between species of same family such as Arctic char and brook trout (salmonidae), haddock and cod (gadidae). Only lysozyme levels showed a marked variation with salinity where seawater fish showed approximately two times higher lysozyme activity than freshwater-reared fish species. Characterization of proteases with specific inhibitors showed Arctic char, brook trout, haddock and cod having higher levels of serine over metalloproteases whereas koi carp and striped bass had higher levels of metalloproteases over serine proteases. In contrast, hagfish had almost equal proportion of both serine and metalloproteases. This study demonstrates variation in the level of hydrolytic enzymes in the epidermal mucus of fish. These results provide preliminary information for a better understanding of the role of epidermal mucus and its components in the fish innate immune system.     

Effect of temperature on proteinase activities in the intestinal chyme and the mucosa of various ecological fish groups

Effect of temperature on activities of proteinases in intestinal chyme and mucosa was studied in three fish species (pike-perch, zope, roach) belonging to different ecological groups by their type of feeding. There was revealed a significant difference of dependence of enzyme activities in chyme on temperature in the benthophage roach (a higher level of relative activity in the zone of lower temperatures and a larger zone of temperature optimum) as well as of values of apparent energy of activation of the protein hydrolysis process as compared with that in plankto- and ichthyophages--zope and pike-perch, which indicates a significant effect of the enteral microbiota proteinases and of nutrition objects on characteristics of hydrolases functioning in fish intestine.     

Seasonal variations in the activities of peroxide metabolism enzymes in erythrocytes of freshwater fish species

Superoxide dismutase, catalase and peroxidase activities were estimated in erythrocytes of the carp, the tench and the crucian carp. The enzymes showed higher activities in spring (1982) than in autumn (1981). In both seasons the highest levels of these antioxidative defense enzymes were found in erythrocytes of the crucian carp.     

Liver gangliosides of various animals ranging from fish to mammalian species

Liver gangliosides of different animal species were analyzed. Bony fish liver contained a major ganglioside that migrated faster than GM3 on thin-layer chromatography (TLC). This ganglioside was identified to be GM4 (NeuAc) by methods including product analysis after sialidase treatment and negative-ion electrospray ionization (ESI)-mass spectrometry (MS). The presence of GM4 (NeuGc) in fish liver was also demonstrated. The main ganglioside band of bovine liver consisted of two different molecular species, i.e. GD1a (NeuAc/NeuAc) and GD1a (NeuAc/NeuGc). Major gangliosides of liver tissue exhibited a distinct phylogenetic profile; GM4 was expressed mainly in lower animals such as bony fish and frog liver, whereas mammalian liver showed ganglioside patterns with smaller proportions of monosialo ganglioside species. While c-series gangliosides were consistently expressed in lower animals, they were found only in mammalian liver of particular species. No apparent trend was observed between the concentration of liver gangliosides and the phylogenetic stage of animals. The present study demonstrates the species-specific expression of liver gangliosides.     

Production of reactive oxygen species (ROS) by various marine fish species during the larval stage

Previous studies have indicated that the devil stinger produces reactive oxygen species (ROS) during early development from fertilized egg to larva. To determine whether ROS generation is a common feature in marine fish species, we conducted chemiluminescence analysis using ROS specific probe (L012) on larvae of six marine fish species. Marbled rockfish, black rockfish, and devil stinger showed higher levels of chemiluminescence response (CR), whereas the levels of CR of sevenband grouper, tiger puffer, and red seabream were fairly lower. These CRs were inhibited by the addition of superoxide dismutase. Hypersensitive photon-counting microscopic observation of black rockfish suggested that ROS production was concentrated in the head area. Our results suggest that the larvae of these six marine fishes produce ROS to considerably different extents depending on species, and that rockfish species, belonging to ovoviviparous fish, tend to produce much higher levels of ROS especially at the later larval stage.     

Performance of a species distribution model based on presence/absence for various population sizes in the amphidromous fish species Sicyopterus japonicus

We formulated a species distribution model (SDM) for the amphidromous fish Sicyopterus japonicus on the basis of presence/absence data and confirmed its performance at various population densities. The best-fit SDM selected several environmental factors, including water depth and velocity, and performed validated prediction of presence/absence of various densities. The predicted probability of occurrence was positively correlated with the observed density. The density was positively related to habitat occupancy, suggesting that this species occupied a wide range of habitats under high densities, but only optimal habitat under low densities. Therefore, the threshold value for predicting presence/absence increased with decreasing densities.     

Relationship between the spike activities of the small and large intestines

Experiments were performed using chronically implanted electrodes on the dog smooth muscle wall of the stomach and of the small and large intestines. Electrical activity of the muscle wall was recorded before and after feeding. When reaching the terminal ileum the active part of the migrating myoelectrical complex (MMC) continuously induced bursts of spike potentials superimposed on the slow waves. This electrical activity spread to the ascending colon. We also showed the existence of a spike activity on the terminal ileum independent of the MMC (appearing during the phase 1) and propagating to the colon. A relationship between the spike activities of the small and large intestines was also present after feeding. Beside the well-known gastro-colic reflex, we observed an increase in the spike activity of the terminal ileum and ascending colon between the 4th-5th hours after feeding. This probably corresponds to the arrival of the first portions of contents, evacuated from the arrival of the first portions of contents, evacuated from the stomach, and of the last portions of small intestinal contents. In conclusion, there is a relationship between the spike activities of the small and large intestines in starving animals and after feeding, and the terminal ileum plays a substantial role in this relationship.     

The effects of environmental temperature on the properties of myofibrillar adenosine triphosphatase from various species of fish

1. Myofibrillar adenosine triphosphatase (ATPase) activities were measured for white myotomal muscle of 19 species of fish. 2. The activity was measured at different temperatures and after periods of preincubation at 37 degrees C. 3. The inactivation half-life at 37 degrees C depended on environmental temperature, increasing as the temperature increased. 4. Cold-water fish had higher myofibrillar adenosine triphosphatase activity at low temperatures than had warm-water fish. 5. The significance of these results is discussed.     

Paraquat and roundup effects on nucleases activities of alfalfa seedlings and alfalfa nucleases activities on paraquat-treated and roundup-treated nucleic acids

The specific activities of acid (pH 5.5) and neutral (pH 7) DNases and RNases were determined in alfalfa (Medicago sativa L.) seedlings grown in the dark in the presence of 3.7 mM paraquat (PQ) or 1 mM roundup (RD). Seedlings were taken at 0, 1, 3, and 5 days. Plant growth parameters (plant height and fresh weight) were dramatically reduced under these conditions of growth comparing to the control (grown in water). The DNase and RNase specific activities of herbicide-treated seedlings were reduced. The reduction of activities ranged by about 50–90 and 15–70% in PQ- and RD-treated seedlings, respectively. In vitro, PQ- and RD-treated nucleic acids [single-stranded DNA (ssDNA), RNA, and plasmid DNA (pl-DNA)] were incubated with acid and neutral nucleases. Both enzymes were isolated and purified from alfalfa seedlings. Electrophoretic analysis on agarose gel of the above incubated mixtures revealed the following: (a) neutral nuclease (pH 7) was capable of hydrolyzing PQ-treated ssDNA while acid nuclease (pH 5.5) was incapable. This could be due to the fact that acid and neutral nucleases displayed different base linkage specificity toward ssDNA; (b) RD formed strong complexes with ssDNA that were unable to be hydrolyzed by both nucleases; (c) in contrast, both enzymes were capable of hydrolyzing PQ- or RD-treated RNA; (d) neutral nuclease was capable of nicking and linearizing both PQ- and RD-treated pl-DNA while acid nuclease had the same activity only toward the PQ-treated pl-DNA; (e) the enzyme activities were not inhibited in the presence of both herbicides. The data suggest that the complexes of PQ or RD with DNA should not be functional substrates of nucleases, and consequently cell processes (e.g., metabolism of nucleic acids, gene expression, replication), in which DNA and nucleases are involved, could be disturbed.     

Calcium transport activities of plasma membranes isolated from the livers of various animal species

1. Plasma membranes of comparable yield and purity were isolated from the livers of various animal species belonging to phylogenetic groups from Amphibia to Mammalia. 2. Calcium transport activity was observed in all liver plasma membranes examined. 3. No phylogenetic pattern of expression of the liver plasma membrane calcium transport system was observed, with the order of activity being: guinea pig greater than rabbit greater than frog greater than chicken = hamster greater than rat = budgerigar = turtle greater than beef cattle greater than mouse = duck. 4. Calcium transport activity was only 9.7 and 8.7% of adult frog levels in plasma membranes isolated from the livers of tadpoles without and with limbs, respectively. 5. Liver plasma membrane calcium transport activity was 25% higher in adult chickens than in day-old chicks. 6. A possible role for thyroid hormone in the development of the liver plasma membrane calcium transport system is discussed.     

除草剂草甘膦异丙胺盐水剂对中华大蟾蜍心电活动的影响

采用生物信号采集处理系统对草甘膦异丙胺盐胁迫条件下中华大蟾蜍(BufogargarizansCantor)的心率和心电活动的相关指标进行了测定和分析。不同浓度的草甘膦异丙胺盐溶液被喷施到中华大蟾蜍的体表,由皮肤进入体内而作用于心脏。试验结果表明:随着草甘膦异丙胺盐处理浓度的增大,中华大蟾蜍的心率减慢,心电图中P波、R波和T波的电压峰值降低,而P-R间期、QRS期和Q-T间期的时值延长。草甘膦异丙胺盐胁迫条件下,蟾蜍心率与心电图的各项指标有显著相关性,可用多元线性回归模型分别对蟾蜍心率与心电图中P、R、T波和P-R、QRS、Q-T间期的相关关系进行拟合。多元回归分析结果表明,蟾蜍心电图中Q-T间期值对心率的影响最大,可以推断草甘膦异丙胺盐主要是通过延长蟾蜍心电活动周期中Q-T间期时值即心室收缩期而延长心动周期,导致蟾蜍心率减慢。由此可见草甘膦异丙胺盐水剂的喷施对中华大蟾蜍心脏的电活动周期和机械活动周期均造成了一定影响。