Purification of lectin from perch (Persa fluviatilis L.) roe specific to cellobiose and study of its characteristics

Two lectins with different carbohydrate specificity were purified from perch (Persa fluviatilis L.) roe (coastal ecological form) by affinity chromatography on ovariomucine H-sepharose from a human ovary cyst. One lectin was eluted by cellobiose and another lectin was eluted by L-fucose. The L-fucose-specific lectin interacted only with L-fucose and its derivatives, but did not interact with cellobiose and salicin. The cellobiose-specific lectin interacted with all the examined carbohydrates, but cellobiose was the best inhibitor. This lectin can be also purified on cellulose as an affinity sorbent. Unlike the L-fucose-specific lectin from perch roe, the cellobiose-specific lectin is less soluble in water-saline solutions. Lectin solubility increases greatly in presence of specific inhibitors, cellobiose, in particular. L-fucose, alpha-methyl-L-fucopyranoside and 4-nitrophenyl-alpha-L-fucopyranoside are equivalent inhibitors for both lectins. According to SDS-PAGE data, the lectins contain two components with molecular weight 12-13 kDa. In solutions, these components form molecules with 50 or 100 kDa (depending on pH). Data obtained from electrophoresis in PAAG in alkaline (pH 8.9) and acidic system (pH 4.3), and SDS-PAGE did not display essential distinctions between these both lectins.     

Temperature dependent characteristics of intestinal glycyl-L-leucine dipeptidase in boreal zone fish

Three kinds of boreal zone fish were investigated for gastrointestinal glycyl-L-leucine (GL) dipeptide cleaving activity as a function of feeding stage and seasonal changes. The enzyme activity tested in the perch (Perca fluviatilis L.) intestine increased steadily during digestion and rapidly disappeared after completion. The temperature characteristics and the seasonal changes in dipeptide cleaving activity in pike perch (Stizostedion lucioperca L.) and bream (Abramis brama L.) were studied. In summer, the maximal activities in the pike perch and the bream were found at temperatures of 40 and 30 degrees C, respectively. In winter, the temperature of maximal activity in pike perch fell to only 30 degrees C, whereas no changes were observed in bream. The activation energies in bream and pike perch were several times lower in winter than in summer. Seasonal changes in the dipeptide cleaving activity at low temperature relative to that at the temperature of maximal activity were found. At high temperatures, the stability of the enzyme decreases in winter and increases in summer, but in the presence of a substrate the thermal stability of the enzyme increases both in winter and in summer. In our experiments, we found that in these fish, GL dipeptidase was unstable at 0 and -10 degrees C.     

Oxygen consumption of Limnomysis benedeni Czerniavsky, 1882 (Crustacea: Mysidacea), a Pontocaspian species in Lake Balaton,Hungary

Hydrobiologia - Limnomysis benedeni Czern. was artificially introduced into Lake Balaton in 1950 in order to increase the food source of young (1+, 2+) pike perch (Stizostedion lucioperca L.). L....     

Study of the haemoglobin content of a single erythrocyte (M index) in various cultured fish species

Investigations of the erythrocyte count, haemoglobin content and M index of 7 fish species were performed under equivalent environmental conditions in the same period of the year. The species were: bighead Hypophthalmichthys nobilis (Richardson), carp Cyprinus carpio L., silver carp Hypophthalmichthys molitrix (Val.), grass carp Ctenopharyngodon idella (Val.), sheat-fish Silurus glanis L. , pike Esox Indus L., pike-perch Lucioperca lucioperca (L.). Bighead and carp are carnivorous and omnivorous, silver carp and grass carp are herbivorous and sheat-fish, pike and pike-perch are predatory. Significant differences were demonstrable between M values of fish living on different type of food, where the M index is: It is concluded that M index values are useful as one parameter differentiating between fish with distinctly different food habits.     

Proteolytic activity and electrophoretic profiles of proteases from seminal plasma of teleosts

Using gelatin-SDS-PAGE, proteolytic activity was found in the seminal plasma of 10 teleosts: common carp Cyprinus carpio , bream Abramis brama , ide Leuciscus idus , chub Leuciscus cephalus , rainbow trout Oncorhynchus mykiss , grayling Thymallus thymallus , perch Perca fluviatilis , pike Esox lucius , goldfish Carassius carassius and pikeperch Stizostendion lucioperca . This activity was also measured, using azoalbumin as a substrate, in the seminal plasma of these species, with exception of pikeperch and goldfish. When azoalbumin-hydrolysing activity was expressed per volume, it was highest in common carp. Otherwise, as expressed per g of protein, the activity was highest in pike. The lowest proteolytic activity (expressed per g and volume) was observed in perch seminal plasma. Using gelatin containing polyacrylamide gels for detecting gelatinolytic activity, species-specific electrophoretic profiles were found. For all cyprinids two similar bands with a molecular mass of 68 and 74 kDa were found. The seminal plasma of grayling and rainbow trout showed similarities in the 41 kDa band. Perch and pikeperch had one similar main band with a molecular mass of 61 kDa. Proteolytic enzymes of seminal plasma from pike showed high individual variability. These results suggest that multiple forms of proteolytic enzymes exist in seminal plasma of teleosts and they differ among fish families and species.     

Induction of apoptosis in populations of mammalian cells in vitro under the influence of lectins

A study of the cytogenetic action of lectins that differ in terms of origin and hydrocarbon specification is presented. In a culture of Chinese hamster cells it is shown that at high concentrations (20 μg/ml), all lectins are capable of inducing an increase in the level of apoptosis two days after treatment with a high degree of reliability. In the case of perch roe lectin, such an increase occurs both immediately after treatment as well as two days later. It is found that, unlike lentil and perch roe lectin, in a culture of human cells black elder lectin does not have any effect on the incidence of apoptotic cells. A trend towards stimulation of proliferation of human cells under the influence of perch roe lectin at low concentrations (0.2 μg/ml) is discovered.     

Environmental correlates of population variables of perch (<Emphasis Type="Italic">Perca fluviatilis</Emphasis>) in boreal lakes

We examined relationships among perch population variable parameters in two types of lakes, lakes with perch (P-lakes, n?=?15) and lakes with perch and roach (PR-lakes, n?=?10) using redundancy analysis (RDA) to relate population variables to environmental factors. Effects from environmental factors were tested for significance by means of permutation tests (Monte Carlo). Three factors, pH, altitude and fraction of roach (by number) in the gill net catches, explaining 47.9% of the variation, had significant effects on perch population variables. The significance of pH was improved by partialing out the effect of conductivity and roach. Similarly, the significance of altitude was improved by partialing out the effect of pH and roach, and the significance of roach was improved by partialing out the effect of pH and altitude. When the fraction of pike in the catch was included in the analysis, the effect of roach was not significant and vice-versa, as roach and pike fractions were correlated with each other. The effect of pike was significant when roach was not included, but the effect was not as strong as the effect of roach. A biplot was constructed by plotting population variables on the first and second RDA axis, with arrows showing five selected environmental factors. Growth of 3+ to 5+ perch was positively related to pH and altitude, perch catch per unit effort was negatively related to pH and altitude, and age specific perch weight was negatively related to fraction of roach. The relationship between growth of 2+ perch and pH was not as strong as the relationship between pH and the growth of older perch. Moreover, the growth of 2+ perch was negatively related to the fraction of roach, probably due to competition between young zooplankton feeding perch and roach.     

Foraging behaviour and capture success in perch, pikeperch and pike and the effects of prey density

The effect of school size on capture success in three different piscivores, perch Perca fluviatilis , pikeperch Stizostedion lucioperca and pike Esox lucius , was investigated. Roach Rutilus rutilus were used as prey in a pool experiment where individual predators were presented prey at densities of one, two, four, eight and 16 prey, respectively. Treatments were replicated seven times for each predator species. Perch was at first virtually unable to capture a prey from a school and suffered a significant confusion effect with increasing prey density. The effect, however, was limited in the long run, as the perch was a very effective predator in its hunting strategy where it singled out and repeatedly attacked single prey irrespective of prey density or school size. Pikeperch and pike were able to attack and capture prey at any prey density equally successfully and thus did not suffer from a confusion effect. Neither did these predators receive any apparent advantages from increasing prey density.     

Reproduction effort versus the environment; case histories of Windermere perch, Perca fluviatilis L., and pike, Esox lucius L.

The theoretical limits of net reproduction rates for perch, Perca fluviatilis L. and pike, Esox lucius L, have been estimated from the upper and lower limits of fecundity, growth, mortality, age of maturity and biomass of parental stock, observed in Windermere over 40 years. Differences in egg numbers due to changes in fecundity were for perch × 1.5, for pike × 2. Assuming the same mortality, a fast growing cohort would produce more eggs during its lifespan than a slow growing, for perch × 2, for pike × 9. Changes in mortality from high to low resulted in more eggs, for perch × 7, and for pike (where high mortality includes natural and fishing mortality) × 10. Change in age at first maturity from 3 to 2 years was unimportant. Biomass of parental stock varied by × 6 for perch and × 3 for pike, and number of eggs laid varied by x2 for perch and × 5 for pike. The number of recruits at age 2 years varied by × 300 for perch and × 7 for pike. It was concluded that temperature in the first summer of life and predation were of major importance in regulating the number of recruits. Ricker recruitment curves and a simple model showed that perch, but not pike, could produce maximum recruitment from minimum parental stock. For perch, but not pike, compensatory density dependent mortality increased with increased parental stock. It was concluded that responses by the perch and pike populations to a changing environment can not be of sufficient magnitude to prevent wide fluctuations in recruitment.     

Studies of oligosaccharide specificity of perch roe fucolectin,using gold labeled neoglycoproteins

The specificity of perch (Perca fluviatilis) roe fucolectin was studied using the protein dot blot technique, followed by detection with colloidal gold-labeled neoglycoproteins bearing human milk polysaccharides. The strongest binding was noted with the H type 1 pentasaccharide lacto-N-fucopentaose (LNFP I, Fuc alpha 1-2 Gal beta 1-3 GlcNAc beta 1-3 Gal beta 1-4Glc); the interaction with the H type 6 trisaccharide 2'-fucosyllactose (2-FL, Fuc alpha 1-2 Gal beta 1-4 Glc) was weaker. Binding of the perch lectin to the Lewis antigens (associated with tumors and embryonic tissues) was also studied. It was found that the lectin weakly interacted with the hexasaccharide lacto-N-difucohexaose I, Fuc alpha 1-2 Gal beta 1-3[Fuc alpha 1-4]GlcNac beta 1-3 Gal beta 1-4 Glc), but not with Lea, Lec, Lex antigens. Thus, perch roe lectin exhibited pronounced differences in carbohydrate specificity from other fucolectins--a feature that may be used in structural studies and isolation of fucose-containing glycoconjugates.     

Effect of temperature on proteinase activities of enteral microbiota and intestinal mucosa of fish of different ecological group

Effect of temperature on proteinases activities of enteral microbiota and of intestinal mucosa was studied in five fish species (roach Rutilus rutilus, crucian carp Carassius carassius, common perch Perca fluviatilis, pike-perch Zander lucioperca, and pike Esox lucius) belonging by the nutrition type to different ecological groups. Essential differences of temperature characteristics of proteinases of intestinal mucosa and of enteral microbiota are revealed in fish belonging by the nutrition type to different ecologic groups. The character of the t0-function of proteinases of intestinal mucosa and enteral microbiota by casein and hemoglobin as a rule is different. The highest values of relative proteinases activities for casein in the zone of low temperatures (38 and 45.3 % of the maximal activity) are found at study of proteinases of enteral microbiota in common perch and crucian carp. The latter indicates a significant adaptability of the enteral microbiota proteinases of common perch and crucial carp to functioning at low temperatures.     

The population dynamics of pike,Esox lucius,and perch,Perca fluviatilis,in a simple predator-prey system

Synopsis The population dynamics and predator-prey relationship of pike, Esox lucius, and perch, Perca fluviatilis, were examined in simple fish communities in two adjacent shallow lakes, Lochs Kinord and Davan, Deeside, Scotland. Few perch survive to age 3 but Z is low for fish > 3 years and perch live up to 17 years. Population fecundity remained relatively high and constant in perch because of the multi-age spawning stock and the presence of older more fecund perch. Growth rates of perch in both lochs are relatively high as a consequence of low stock abundance. The N, B, and P of adult perch were unusually low. The age range of pike, and N, B, P, and growth were in the range of values reported elsewhere. There was little variation in the strength of pike year classes and the importance of cannibalism and low occurrence of alternative prey in the lochs suggest that the populations were self-regulating. Cannibalism by adults was responsible for most of mortality in perch larvae, and predation by pike and adult perch was responsible for the majority of juvenile losses. This conclusion is supported by the high biomass ratios of pike:juvenile perch of 1.0–30.8. While the number of adult fish was almost equal, the biomass of adult pike was 2–3 × that of perch in Kinord and 6 × in Davan. In L. Kinord, where year class strength was stable, high predation pressure from perch and pike reduced perch abundance rather than eliminated year classes. Perch year classes fluctuated in abundance in L. Davan and were eliminated in the first summer in two sampling years. The pike, and particularly the perch populations, have features characteristic of fish communities in unperturbed ecosystems: namely, a wide range of age classes, stability in biomass with variation dampened by longevity, and low production.     

Consumption of crucian carp (Carassius carassius L., 1758) by restocked pike (Esox lucius L., 1758) in a lake with frequent winter hypoxia

To assess the effects of stocked pike (Esox lucius L.) on crucian carp (Carassius carassius L.) biomasses, the annual consumption of pike was estimated and compared with removal fishery catches. The studied lake, Lake Savijärvi, is a small (40 ha), shallow and eutrophicated lake in southern Finland with frequent algal blooms during summers and fish kills during winters. Until the 1980s, the fish fauna consisted of pike, perch (Perca fluviatilis L.), roach [Rutilus rutilus (L.)], crucian carp, and tench (Tinca tinca L.). Since 2003, when crucian carp were abundant in the extreme and with only a few roach and tench individuals, the lake has been biomanipulated by removal seining. To enhance the effects of seining, piscivorous pike were restocked in the spring of 2008. During the 3‐year study, the catch of crucian carp decreased from 243 to 136 kg while the catch of pike increased from 0.1 to 5.7 kg per seining hectare due to their spawning in the lake as of 2009. The total crucian carp consumption by captured pike was 588 kg during the 2008–2010 study period. When applying the number of pike estimated with the mark‐recapture method, consumption estimate increased to 917 kg, or 22.9 kg per lake hectare in 2010. Thus, pike consumption of crucian carp was about 17% of the removal catch in 2010.     

Purification of lectin from fruiting bodies of Lactarius rufus (Scop.: Fr.)Fr. and its carbohydrate specificity

A lectin from fruiting bodies of Lactarius rufus (Scop.: Fr.)Fr. has been purified by affinity chromatography on copolymer of polyvinyl alcohol with a blood group B specific substance. The lectin gives a single band at disk-electrophoresis in acidic (pH 4.3) and alkaline (pH 8.6) buffer systems. Under electrophoresis in 10-20% SDS-PAGE, the lectin consists of identical subunits with molecular weight 17 +/- 1 kDa. Molecular weight of the lectin is 98 kDa according to gel-chromatography on Tojopearl HW-55. It is supposed that the lectin contains six subunits. The lectin is quite enough stable in pH 4.0-10.0, its activity does not depend upon bivalent metal ions. When heating the lectin solution to 65 degrees C it lost more than 85% of its activity. The lectin agglutinates human etrythrocytes without any marked group specificity, it agglutinates 2-4 times worse rabbit erythrocytes, very weakly crucian erythrocytes and does not agglutinate sheep erythrocytes. Mono- and disaccharides are not inhibitors of the lectin activity, while alpha-phenyl-N-acethyl-D-glucosaminopyranosid (0.08 mM) and 4-nitrophenyl-beta-D-glucosamin are the best inhibitors of its activity. Among glycoproteins the best inhibitors of the lectin activity are: group-specific substances from human blood erythrocytes, asialosubmaxillary bovine mucin, human and bovine thyroglobulin and more weak inhibitors are fetuin, transferrin and human Ig G.     

Purification and characterization of a new lectin from the hard roe of skipjack tuna,Katsuwonus pelamis

Fish eggs are known as a rich source of lectins. In this study we purified and characterized a lectin from unfertilized Katsuwonus pelamis hard roe. K. pelamis lectin (KPL) was purified by separation into two fractions above and below the molecular weight of 10kDa using ultramembrane, gel filtration on a Sephadex G-100, and affinity chromatography on an asialofetuin-Sepharose 4B. KPL is a glycoprotein of 140kDa, composed mainly of aspartic acid, glycine, phenylalanine, glutamic acid, threonine and serine residues. Analysis of the carbohydrate composition by gas-liquid chromatography indicated that carbohydrates constituted 14% of the total weight and this 14% is comprised of mannose, galactose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, fucose, arabinose and sialic acid. The lectin is comprised of four subunits. These subunits have a molecular mass corresponding to 35kDa. KPL specifically agglutinated human blood type A erythrocytes and, in a hemagglutination inhibitory test, the potent inhibitors were D-galactose, lactose, lactosamine, asialofetuin, N-acetyl-D-galactosamine, O-serinyl-2-acetamido-2-deoxy-alpha-D-galactopyranoside and O-serinyl-2-acetamido-2-deoxy-beta-D-galactopyranoside (O-serinyl-beta-D-GalNAc). The first 10 residues of the N-terminal region were determined as PVELCDAKCT. Furthermore it was determined that the hemagglutinating activity of KPL was dependent on divalent metal cations and that the optimum activity of KPL was exhibited at 40 degrees C and pH 6.0-8.5 in the presence of Ca2+.     

Mark-recapture experiments on fish in Windermere, 1943–1982

Marking experiments on perch, Perca fluviatilis L., pike, Esox lucius L., and charr, Salvelinus alpinus L., have been carried out in Windermere. Numbered tags were used for individual identification, except in some short term experiments on charr. A total of 13 182 perch and 4696 pike were tagged; 2066 charr were tagged subcutaneously and marked with a fin clip. In the short-term experiments 2015 charr were marked with a fin clip or punched hole. Perch were recaptured up to 8 years after tagging, pike 12 years and charr 5 years. Estimates of numbers and mortality were not satisfactory, except for limited sections of the populations of pike and charr. The main reasons for the unsatisfactory results were: for perch, unknown mortality at time of tagging; for pike, selectivity of the gillnets and in some years low numbers tagged; for charr, restriction to few spawning sites and in some places low numbers recaptured. The experiments provided useful information on movements and growth, which could not have been obtained in any other way.     

Stunted growth of pikeperch Sander lucioperca in Lake Sahajärvi, Finland

Growth of pikeperch Sander lucioperca in the eutrophic and clay-turbid Lake Sahajärvi, Southern Finland, was extremely slow in comparison with other lakes at similar latitudes. The most important food item in July was phantom midge larvae Chaoborus flavicans for all sizes of S. lucioperca (239–423 mm total length L _T), while later, in August and September, the diet of S. lucioperca (149–407 mm L _T) consisted of small (30–100 mm L _T) perch Perca fluviatilis , ruffe Gymnocephalus cernuus and roach Rutilus rutilus .     

Studies on lectins. LVII. Immunofluorescence localization of lectins present in fish ovaries

The occurrence of endogeneous lectins in the ovaries of four fish species has been studied by indirect immunofluorescence staining with antibodies against individual lectins. Paraffin sections of the ovary of perch (Perca fluviatilis L.) were treated with an antibody against perch lectin. In cryostat sections of the tench (Tinca tinca L.) ovary, the L-rhamnose-specific lectin "I" was detected with a specific antibody. In cryostat sections of both roach (Rutilus rutilus L.) and rudd (Scardinius erythrophthalmus L.) ovaries, lectins were localized using a single antibody against roach lectin. The isolation of tench lectins is briefly described. In the fish species employed for this study, lectins are associated exclusively with the content and surrounding membrane of cortical vesicles situated within the cytoplasm of maturing oocytes. The positive reaction with lectin antibody was observed almost immediately after the formation of the first cortical vesicles in the peripheral cytoplasm of early previtellogenic oocytes. Their lectin content increases during the later stages when cortical granules fill the whole cytoplasm before moving towards the cell periphery, as the oocyte starts to accumulate yolk. The presence of lectins within cortical vesicles is significant also in view of the polysaccharide content of these structures. In the vitellogenic oocytes lectins seem to move towards the cell periphery and accumulate beneath the plasma membrane. Our observations are discussed in view of the present ideas on the intracellular function of lectins, and with respect to the role of cortical vesicles in fertilization and in post-fertilization modifications of the egg envelopes.     

A new lectin with highly potent antihepatoma and antisarcoma activities from the oyster mushroom Pleurotus ostreatus

A dimeric lectin, composed of subunits with a molecular weight of 40 and 41 kDa, respectively, and demonstrating similarity in N-terminal sequence to each other and to Aleuria aurantia lectin, was isolated from fresh fruiting bodies of the edible mushroom Pleurotus ostreatus. The lectin was unadsorbed on DEAE-cellulose in 10 mmol/L phosphate buffer (pH 7.5), subsequently adsorbed on CM-cellulose in 10 mmol/L NH(4)OAc (pH 5.4), and came off in the first peak from a Superose 12 column during fast protein liquid chromatography. The lectin was acid-labile, alkali-labile, and heat-labile. Its hemagglutinating activity was sensitive to inhibition by CaCl(2), MgCl(2), MnCL(2) and FeCl(3) and potentiation by AlCl(3). Melibiose, lactose, d-galactose, alpha-methyl-d-galactopyranoside, N-acetylneuraminic acid, raffinose, and inulin were capable of inhibiting its hemagglutinating activity, with melibiose being the most potent. The lectin exerted potent antitumor activity in mice bearing sarcoma S-180 and hepatoma H-22. Survival in these mice was prolonged and body weight increase reduced after lectin treatment.     

Purification and study of carbohydrate specificity of lectin from Sarcoscypha coccinea (Fr.) Lambette

A lectin that revealed affinity for lactose, N-acethylactosamine, 4-nitrophenyl-beta-D-gluco- and galactopyranosides from fruiting bodies of Sarcoscypha coccinea (Fr.) Lambette was purified by affinity chromatography on immobilized ovomucoid. According to electrophoresis data in 15% SDS-PAGE the lectin contains two very low-differing components with molecular weight 32 kDa. Molecular weight of the lectin is 128 kDa according to gel-chromatography on sephadex G-200. The lectin agglutinates rabbit erythrocytes and slightly weaker agglutinates human erythrocytes. After dialysis against 1% EDTA sodium salt solution the lectin loses hemaglutinating activity, but after the next dialysis against CaCl2 solution it is restored.