The adaptation of digestive enzymes to temperature, season and diet in roach, Rutilus rutilus and rudd Scardinius erythrophthalmus; Proteases

The seasonal changes of proteolytic activity in the gut content of roach R. rutilus and rudd S. erythrophthalmus in four Tyrolean lakes and the adaptation of the proteases to constant temperatures and different natural diets were studied. In rudd proteolytic activity remains nearly constant throughout the year. In roach proteolytic activity increases and then decreases during the first three months after the thaw in spring. This period appears to be endogenously controlled and is followed by a second stage in which proteolytic activity is influenced by environmental factors. Under natural conditions both species have a higher proteolytic activity when feeding on animals than on detritus. Under laboratory conditions there is a slight maximum at an environmental temperature of 16° C when the fish had been fed on meal worms. With plant food a significant dependence of proteolytic activity on environmental temperature was only found in roach.     

The adaptation of digestive enzymes to temperature, season and diet in roach Rutilus rutilus L. and rudd Scardinius erythrophthalmus L. Cellulase

The cellulolytic activity of the contents of the intestinal tract of populations of roach and rudd in four Tyrolean lakes was studied. Much of the activity was taken up with the diet. In fish feeding on plants and detritus a lower cellulolytic activity was found than in fish feeding on zooplankton or on arthropods. A seasonal pattern of cellulolytic activity was demonstrated with a peak in the autumn. It is assumed that the cellulolytic activity found in the intestinal contents of cyprinids is insufficient for the digestion of fibre, but that is suffices—particularly in omnivorous species—to support the breakdown of cell walls.     

Electrophoretic identification of roach (Rutilus rutilus L.), rudd (Scardinim erythrophthalmus L.), bream (Abramis brama L.) and their natural hybrids

Roach, rudd, bream and their natural hybrids of 2 cm standard length or larger can be definitively identified by their enzyme electrophoretic patterns. Zymograms of lactate dehydrogenase and esterases as produced by vertical starch gel electrophoretic analysis of whole fry or adult eye extracts are the most useful in this respect. The lactate dehydrogenase isozymes, containing B sub-units, migrate more anodally in rudd and bream than in roach. Due to the tetrameric structure of lactate dehydrogenase, in hybrid rudd x roach and roach x bream, eleven isozymes can be observed as compared with six in the parental patterns. Esterases show unique patterns for all species and hybrids. With the exception of one fraction in rudd x bream, the esterase patterns of hybrids show summations of the parental phenotypes.     

Amylolytic activity in fish intestinal mucosa: temperature effects

The activity, temperature characteristics and energy of activation of amylolytic enzymes in the intestinal mucosa were studied in six species of fish living in a boreal zone [burbot (Lota lota L.), northern pike (Exos lucius L.), perch (Perca fluviatilis L.), bream (Abramis brama L.), roach (Rutilis rutilis L.), and carp (Cyprinus carpio L.)] and in three species from tropical and subtropical areas [pilchard (Sardina pilchardus W.), jack mackerel (Trachurus trecae C.) and round sardinella (Sardinella aurita V.)]. The amylolytic activity correlated with the feeding habits: it was essentially lower in predators. The enzyme activity at low temperature, relative to the maximal activity, was correlated with the natural environmental temperature where the species lived. At low temperature the relative activity was higher in boreal fish than in tropical and subtropical fish. We found a breakpoint in the Arrhenius plots in all fish species, except for jack mackerel. The energy of activation in predators decreased below the breakpoint in the low-temperature region. The energy of activation in benthophages of the Aral-Ponto-Caspian area was lower at higher temperatures above the breakpoint. A reduction in activation energy in the range of physiological temperatures might indicate adaptation to the environmental temperature.     

The influence of simulated aquatic macrophytes on the zooplankton consumption rate of juvenile roach, Rutilus rutilus, rudd, Scardinius erythrophthalmus, and perch, Perca fluviatilis

Laboratory experiments showed that, of zooplanktivorous roach, rudd and perch, in the absence of any environmental structure roach were the most efficient feeders, but high densities of simulated submerged marcrophytes elevated perch to this position; rudd matched the performances of perch and roach only at intermediate structure densities. These changes in efficiency rankings with increased structure were the result of overall decreases in the performances of roach and rudd. Simulated water lilies and emergent reeds had their own influences on consumption rate. These effects were largely mediated through relationships between structure density and swimming speed; roach and rudd, not perch, showed reduced swimming speeds in the more structured environments. Observations of the routine behaviour of the fish suggest that their assessment of predation risk may be an important factor in determining their activity level and hence foraging strategy.     

Influence of diurnal rhythms of feeding during intestine total amylolytic activity and activity of alkaline phosphatase in juvenile fish

The total amylolytic activity and activity of alkaline phosphatase in the intestine mucosa of larvae and fries of roach, blue bream, and perch change both during the process of individual development and during the day. Maximal intensities of juvenile feeding was observed primarily in the morning and evening hours. The pattern of diurnal alkaline phosphatase activity correlates to a greater extent to intensity of juvenile feeding, in comparison to the pattern of total amylolytic activity. In planktivorous blue bream, such regularity is more pronounced than in benthivorous roach. The total amylolytic activity in fries of roach, blue bream and perch correlates to fish type of feeding.     

Interspecific dominance amongst three species of coarse fish in Slapton Ley, Devon

Marked changes in the relative numbers of roach Rutilus rulilus (L.), rudd Scardinius erylhroplhalmus (L.) and perch Perca fIuviatilis L. , in Slapton Ley, Devon, have been observed over the period 1967–1978. Historically, the lake had been dominated by rudd and perch; significant numbers of roach were not thought to be present until 1967. From then onwards the roach population exhibited a considerable expansion, apparently replacing the rudd, which had virtually disappeared by 1974. In 1975 extensive mortalities of roach were caused by the pseudophyllidean cestode Ligula intestinalis L. During 1976 and 1977 larger number of rudd, belonging to the 1975 and 1976 year classes, were present, suggesting that the roach mortalities might be giving the rudd a chance to recover. However, very few rudd were present during 1978, possibly because lower incidences of ligulosis during 1976–1978 had allowed the roach to recover from the main outbreak of the disease in 1975, and to replace the rudd for the second time. Evidence from trap catches indicated that the perch population had shown a consistent decline between 1970 and 1977. The major phase of this decrease occurred between 1971 and 1973, when the roach population was increasing the most rapidly. Comparisons were made with changes in fish populations in other localities. The major reason for the changes observed was thought to be a competitive relationship between the plankton feeding younger stages, with roach being the dominant species, although other factors may also have been involved.     

Polymorphism of A, B and C loci of lactate dehydrogenase in European fish species of the Cyprinidae family

In 24 fish species of the Cyprinidae family, belonging to 21 genera, isoenzyme patterns of lactate dehydrogenase (LDH) were determined, which could be classified in the majority of cases into 3 main groups. Isoenzyme patterns in natural hybrids of roach and rudd, roach and bream, roach and bleak were also analysed. In bitterling, polymorphism was observed in B locus of LDH. In white bream polymorphism exists in the A locus. In bream, rudd, silver carp and barbel polymorphism was found in C loci. Isoenzyme patterns indicate that in each case the polymorphism is genetically controlled by two alleles at a single locus. The populations investigated were in Hardy-Weinberg equilibrium. No significant differences were found in the activity of liver LDH in various polymorphic types of C loci of bream and rudd.     

Dung exploitation by the dung beetle Typhaeus typhoeus (Col., Geotrupidae)

Summary The competitive interactions between roach (Rutilus rutilus) and rudd (Scardinius erythrophthalmus) were investigated in two habitats, the open water and the waterlily zone. The growth rates of both species in enclosures were lower when confined together than when alone, demonstrating interspecific competition. Allopatric roach had the highest growth rate in both habitats although the open water was preferred. The diets of both species were dominated by zooplankton in both the open water and waterlily zones. In laboratory experiments, roach had significantly higher feeding rates than rudd when fed D. magna and Cyclops sp. The impact of roach on the mean sizes and densities of zooplankton in the enclosures, together with the results from the laboratory study, indicate that roach were competitively superior in the open water. Because of high mortality in the sympatric waterlily enclosure, no conclusions about interspecific competition in this habitat could be drawn. The observed habitat segregation between roach and rudd was at least partly interactive.     

A search for introgressive hybridization in the rudd, Scardinius erythrophthalmus (L.), and the roach, Rutilus rutilus (L.)

Rudd, roach and their hybrids from an interbreeding population at Norwich were examined electrophoretically for seven enzymes; an examination of pharyngeal teeth supplemented the data. The rudd were compared to rudd from Lake Trawsfynydd, Wales, a lake free from roach. No differences were observed between the rudd from the two localities and no evidence was found of introgression within the fish from Norwich.     

Production, purification, and characterization of alpha-amylase from Thermomonospora curvata.

Thermomonospora curvata produces an extracellular alpha-amylase. Maximal amylase production by cultures in a starch-mineral salts medium occurred at pH 7.5 and 53 degrees C. The crude enzyme was unstable to heating (65 degrees C) at pH 4 to 6, and was activated when heated at pH 8. The enzyme was purified 66-fold with a 9% yield and appeared homogeneous on discontinuous gel electrophoresis. The pH and temperature optima for activity of the purified enzyme were 5.5 to 6.0 and 65 degrees C. The molecular weight was calculated to be 62,000. The Km for starch was 0.39 mg/ml. The amylolytic pattern consisted of a mixture of maltotetraose and maltopentaose.     

Production, purification, and characterization of alpha-amylase from Thermomonospora curvata.

Thermomonospora curvata produces an extracellular alpha-amylase. Maximal amylase production by cultures in a starch-mineral salts medium occurred at pH 7.5 and 53 degrees C. The crude enzyme was unstable to heating (65 degrees C) at pH 4 to 6, and was activated when heated at pH 8. The enzyme was purified 66-fold with a 9% yield and appeared homogeneous on discontinuous gel electrophoresis. The pH and temperature optima for activity of the purified enzyme were 5.5 to 6.0 and 65 degrees C. The molecular weight was calculated to be 62,000. The Km for starch was 0.39 mg/ml. The amylolytic pattern consisted of a mixture of maltotetraose and maltopentaose.     

Ontogenetic changes in haemoglobins in roach, Rutilus rutilus (L.) and rudd, Scardinius erythrophthalmus (L.)

The haemoglobins and globins of the roach, Rutilus rutilus (L.) and the rudd, Scardinius erythrophthalmus (L.) have been studied by starch gel electrophoresis. At all stages of development studied, the roach and rudd haemoglobins give a similar electrophoretic pattern, the young fish of about 2 cm standard length possessing three separate haemoglobins, of which two bands only persist in the adult. The evolutionary significance of multiple haemoglobins in fishes is briefly discussed.     

The Utilization of Cotyledonary Reserves in Phaseolus vulgaris L. cv. Carioca: II. THE EFFECTS OF 6-BENZY LADENINE AND GIBBERELLIC ACID UPON EMBRYONATED AND DETACHED COTYLEDONS

The presence of the embryo axis promotes starch hydrolysis incotyledons of Phaseolus vulgaris and, although reduced ratesof enzyme activity proceed in its absence, weight loss fromthe cotyledon is only significant in its presence. Applicationof gibberellic acid at various concentrations had no effectupon amylase or protease activity in either detached or embryonatedcotyledons: this was not the case for 6-benzyladenine whichpromoted both. In detached cotyledons amylolytic activity wasdirectly proportional to the concentration of applied 6-BA;concentrations below 10^–6 M were unable to completelysubstitute for the presence of the axis, while, above 10^–6M, hydrolytic activity higher than that of treated embryonatedcotyledons was observed. Increasing the concentration of 6-BAhad no effect, however, upon the amylolytic activity of embryonatedcotyledons. Detached cotyledons showed an increased chlorophyllcontent and 6-BA treatment further increased this; the proportionsof chlorophyll a and chlorophyll b remained unaltered. An inhibitor of amylase activity, destroyed by heating, is presentin embryonated cotyledons and can depress amylolysis in detachedcotyledons which appear to be inhibitor-free. The results intotal suggest that the developing axis may regulate reservehydrolysis in three distinct manners: firstly, by stimulatingthe synthesis of amylase via hormonal control, most probablya cytokinin; secondly, by checking the rate of amylolysis bymeans of an amylase inhibitor; and, thirdly, by being a sinkfor the products of reserve degradation.     

Species-specific responses of planktivorous fish to the introduction of a new piscivore: implications for prey fitness

1. Antipredator behaviour by the facultative planktivorous fish species roach (Rutilus rutilus), perch (Perca fluviatilis) and rudd (Scardinius erythrophthalmus) was studied in a multi‐year whole‐lake experiment to evaluate species‐specific behavioural and numerical responses to the stocking of pikeperch (Sander lucioperca), a predator with different foraging behaviour than the resident predators large perch (P. fluviatilis) and pike (Esox lucius). 2. Behavioural responses to pikeperch varied greatly during the night, ranging from reduced activity (roach and small perch) and a shift in habitat (roach), to no change in the habitat use and activity of rudd. The differing responses of the different planktivorous prey species highlight the potential variation in behavioural response to predation risk from species of similar vulnerability. 3. These differences had profound effects on fitness; the density of species that exhibited an antipredator response declined only slightly (roach) or even increased (small perch), whereas the density of the species that did not exhibit an antipredator response (rudd) decreased dramatically (by more than 80%). 4. The maladaptive behaviour of rudd can be explained by a ‘behavioural syndrome’, i.e. the interdependence of behaviours expressed in different contexts (feeding activity, antipredator) across different situations (different densities of predators). 5. Our study extends previous studies, that have typically been limited to more controlled situations, by illustrating the variability in intensity of phenotypic responses to predators, and the consequences for population density, in a large whole‐lake setting.     

Host-constrained epidemiology of the fish tapeworm Ligula intestinalis (L.)

Investigations into the biology of the roach, Rutilus rutilus (L.), and Ligula intestinalis (L.) populations at Slapton Ley, Devon were carried out between October 1982 and December 1984, and additional data collected from the lake since 1977 have been re-analysed. The Ligula population exhibited some unusual features: a limited specificity, a persistently low abundance and a scarcity of multiple infections. The population dynamics of the roach were also unusual in that the individual growth rate was one of the fastest in Britain, the survival was very poor and the year classes tended to alternate in strength. Whether the unusual epidemiology of Ligula could be explained by the unusual population dynamics of the roach is investigated. The transmission period of Ligula to the roach was limited to their first few months of life, probably due to a limited period of feeding on copepods by the fry. This narrow transmission window was almost certainly the major factor that has prevented the abundance of Ligula plerocercoids attaining high levels in the fry or of increasing in subsequent years, and has resulted in their abundance being determined entirely by transmission events to the fry. The limited transmission period has also caused the life-cycles of the roach and Ligula populations to become synchronized. At other localities, the abundance of Ligula is typically high, and can have a significant effect on the mortality of the host population, but, with the low abundance at Slapton, Ligula-induced roach mortality was insignificant. The rapid growth of the roach fry was the most likely explanation for the limited period of feeding on copepods, and the large size, short life-span and low abundance of the roach probably also constrained the build-up of the Ligula population in the lake. The random frequency distribution and scarcity of multiple infections was not considered unusual in view of the very low plerocercoid abundance. The very low abundance of Ligula, the separation of the spawning of roach and rudd in time and space, and the scarcity of rudd may account for the absence of infection in this species. It was concluded that the Ligula population in Slapton Ley was being constrained by the atypical population dynamics of the roach, and not vice versa as theory predicts.     

Impact of polychlorinated biphenyls on the activity of intestinal proteinases and carbohydrases in juvenile roach Rutilus rutilus (L.)

The chronic effects that polychlorinated biphenyls (PCBs) have on the activities of proteinases and carbohydrates in intestinal mucosa and chyme in juvenile roach Rutilus rutilus (L.) have been studied for the first time. Upon consuming food with PCB 50.8 ng/g wet weight for 218 days, the exposition of fingerlings in aquariums with dirt bottoms (contents PCB 425.6 ng/g dry weight) reduces the proteolytic activity of mucosa to a greater degree than amylolytic activity; the activity of sucrase changes in different directions. In 2-year-old fish receiving PCB only with food (50.8 ng/g wet weight bottom), proteolytic and amylolytic activities of the mucosa are reduced 18 and 35% in comparison with the control (52 days); the sucrose activity is reduced 13% (52 days) and 22% (169 days). In most cases the relation of amylolytic activity to proteolytic activity in fish of the experimental groups is less than in the control. The enzyme activities in chyme change in different directions, depending on the terms and conditions of the experiment.     

In vitro analysis of the digestive enzymes amylase and alpha-glucosidase in the midguts of Locusta migratoria L. in response to the myosuppressin, SchistoFLRFamide

We have investigated the effect of the locust myosuppressin, SchistoFLRFamide, on the activity of amylase and alpha-glucosidase in the midgut of 2-week old male locusts. Total enzyme activity in the lumen contents and tissue extracts of midguts responds to SchistoFLRFamide in a dose-dependent manner that appears to vary with the feeding state of the locust and duration of exposure to the peptide. Starvation for 24h prior to assessment alters the distribution of enzyme activity between the midgut lumen contents and tissue extracts in response to SchistoFLRFamide when compared with fed locusts. Duration of exposure to SchistoFLRFamide also alters the distribution of total amylase and alpha-glucosidase activity; as duration of exposure increases, lower concentrations of SchistoFLRFamide increase total enzyme activity in the lumen contents while decreasing total enzyme activity in the tissue extracts. We suggest that the minimum amino acid sequence in SchistoFLRFamide necessary to increase both amylase and alpha-glucosidase activity is DHVFLRFamide. We have determined that two other peptides endogenous to the locust, AFIRFamide and GQERNFLRFamide, increase amylase and alpha-glucosidase activity in midgut lumen contents.     

Cloning and nucleotide sequence of the maltopentaose-forming amylase gene from Pseudomonas sp. KO-8940.

The gene coding for the maltopentaose-(G5)-forming amylase of Pseudomonas sp. KO-8940 was cloned into Escherichia coli and its nucleotides were sequenced. It was expected that a long open reading frame composed of 1,842-bp that encoded 614 amino acid residues for secretory precursor polypeptide including the typical signal sequence with an NH2-terminal was the gene. An extract of Escherichia coli carrying the cloned G5-forming amylase gene had amylolytic activity with which produced only G5 from starch, the same as that of the donor strain enzyme. In the deduced primary structure of this enzyme, the four conserved regions of many alpha-amylases were found, and the COOH-terminal portion of this enzyme showed high homology with other raw starch digesting amylases.     

On the populations of roach (Rutilus rutilus), rudd (Scardinius erythrophthalmus), and their hybriin Esthwaite Water, with notes on the distinctions between them

Samples of cyprinid fishes from Esthwaite Water, Westmorland, have been examined and identified as roach ( Rutilus rutilus ), rudd ( Scardinius erythrophthalmus ), with a high degree (37%) of hybridization between these species. The distinguishing features of roach and rudd are discussed in the light of the difficulty of distinguishing the roach x rudd hybrid from the roach. It is concluded that the only reliable characters are the number and external morphology of the pharyngeal teeth.