Cyprinid assemblages,and the physical and chemical characteristics of small northern Ontario lakes

Synopsis The cyprinid species, and physical and chemical characteristics were recorded from 58 small lakes in the Algoma district of northern Ontario. A group of typical stream-dwelling lithophilous species, which included common shiner, Notropis cornutus, creek chub, Semotilus atromaculatus, and blacknose dace, Rhinichthys atratulus, tended to occupy lakes of larger than average drainage areas. Phoxinus spp., pearl dace, Semotilus margarita, and fathead minnow, Pimephales promelas, occurred commonly in lakes with small drainage areas, and the average drainage area of lakes they occupied was near or below the overall average. Lakes without stream cyprinids were discriminated from lakes with them on a combination of chemical variables, including pH, alkalinity and calcium, which indicated that they were more susceptible to anthropogenic acidification.     

An intergeneric hybrid,Campostoma anomalum × Rhinichthys atratulus,from the Youghiogheny River drainage,West Virginia

The first record of a natural intergeneric hybrid of Campostoma anomalum × Rhinichthys atratulus is described from the Youghiogheny River drainage. Morphometric and meristic characters of the parents and hybrid were used to calculate a mean hybrid index value of 49.8. Dentition of the hybrid was intermediate as was intestinal coiling. In addition to the hybrid index, C. anomalum, R. atratulus and the hybrid were examined electrophoretically. Of the 32 enzymatic loci resolved, six (ADH-A, EST-B, LDH-A, PGM-A, SDH-B and SOD-C) showed hybrid inheritance patterns identical to the enzyme pattern obtained by mixing tissue extracts composed of the two parental species.     

Isozyme expression in F1 hybrids between carp and goldfish

Interspecific genetic differences in malate dehydrogenase (MDH), lactate dehydrogenase (LDH), superoxide dismutase (SOD), and esterase (EST) isozymes in carp (Cyprinus carpio) and goldfish (Carassius auratus) were used to examine the allelic expressions in the hybrid between these species. A unique liver SOD and muscle LDH phenotype unambiguously identifies all presumed hybrid individuals. There was no evidence of F₂ or backcross phenotypes in hybrid individuals. Liver MDH and EST phenotypes in hybrids show a preferential expression of goldfish isozymes. Variation in the levels of carp liver MDH isozymes may result from the polymorphism of a regulatory mutation affecting isozyme expression, leading to gene silencing after duplication.This work was supported through NSERC (Canada) grants to James P. Bogart and John F. Leatherland.     

Malate/lactate dehydrogenase in mollicutes: evidence for a multienzyme protein

The malate (MDH) and lactate (LDH) dehydrogenases belong to the homologous class of 2-ketoacid dehydrogenases. The specificity for their respective substrates depends on residues differing at two or three regions within each molecule. Theoretical peptide-mass fingerprinting and PROSITE analysis of nine MDH and six LDH molecules were used to describe conserved sites related to function. A unique LDH is described which probably also confers MDH activity within the 580 kbp genome of Mycoplasma genitalium (class: Mollicutes). A single hydrophilic arginine residue was found in the active site of the M. genitalium LDH enzyme, differing from an hydrophobic residue normally present in these molecules. The effect of this residue may be to alter active site substrate specificity, allowing the enzyme to perform two closely related tasks. Evidence for a single gene affording dual enzymatic function is discussed in terms of genome size reduction in the simplest of free-living organisms. Since Mollicutes are thought to lack enzymes of the tricarboxylic acid cycle that would otherwise bind and interact with MDH in bacterial species possessing this pathway, active site modification of M. genitalium LDH is the sole requirement for MDH activity of this molecule. The closely related helical Mollicute, Spiroplasma melliferum, was shown to possess two distinct gene products for MDH/LDH activity.     

Lactate and malate dehydrogenase in the fan-shell associated shrimp, Pontonia pinnophylax (Otto): Effects of temperature and urea

In Pontonia pinnophylax (Otto), a crustacean decapod inhabiting the mantle cavity of Pinna nobilis L. (Bivalvia: Pteriomorpha), the lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) activity, and their electrophoretic patterns, were compared in relation to heat and urea inactivation. Activity was higher in LDH than in MDH, and the electrophoretic patterns showed a predominance of LDH-A4 and the presence of both mitochondrial and cytosolic MDH. Heat incubation reduced both enzymatic activities, but more MDH. Also all isozymes showed different heat sensitivity, with anodic forms more heat-resistant than cathodic ones, either in LDH as in MDH. Urea treatment caused also a higher inactivation of the most cathodic isozymes, but MDH appeared more resistant than LDH at 2 M urea. The high polymorphism of these enzymes suggests an adaptation of Pontonia metabolism to hypoxic conditions; moreover, the different isozyme stability grade should be functional to contrast environmental variability.     

The development of lactate and malate dehydrogenases in the C57BL-6 mouse kidney

The development of lactate dehydrogenase (LDH; EC 1.1.1.27) and malate dehydrogenase (MDH; EC 1.1.1.37) was measured in the kidney of male and female $\text{C57BL}\text{6}$ mice from ages prenatal 16 days to 80 days. Maximum reactions rates of the enzymes were measured in vitro by following the reduction of the nicotinamide-adenine dinucleotide spectrophotometrically.Analysis of variance showed no significant sex difference for LDH and MDH. There was a significant sex difference for the ratio LDH:MDH and a significant age difference for LDH, MDH, and the ratio LDH:MDH. In the male and female, LDH activity increased from prenatal 16 days to 30 days. Malate dehydrogenase activity reached adult values at 22 days in the male and at 30 days in the female. The ratio LDH:MDH in the male decreased from prenatal 16 days to 3 days, after which the ratio continued to decline to 20 days at a less rapid rate. This general pattern was also found in the female followed by a further decline in the ratio at 50 days.The development of LDH and MDH in the $\text{C57BL}\text{6}$ mouse is tissue specific and probably parallels the development of the tissue's function. In the case of the kidney, LDH and MDH development may reflect maturation of mitochondrial function and the kidney's ability to concentrate urine.     

Enzyme patterns in two species of Xenopus and their hybrids

Differences in the isozyme patterns of Xenopus laevis and Xenopus mulleri have been utilized to examine the expression of alleles of both species in hybrid animals. Mitochondrial MDH and tetrazolium oxidase phenotypes were examined during the development of non-hybrid embryos of each species and of reciprocal hybrids. Early stages of the hybrids resemble the enzyme phenotype of the maternal parent. Appearance of paternal enzyme takes place just prior to the active feeding tadpole stage for both mitochondrial MDH and oxidase. The maternal effect disappears shortly thereafter in early feeding tadpoles, at which point reciprocal hybrids have identical isozyme patterns. There is no evidence for a predominance of one species over the other. Examination of feeding tadpoles and adult toads indicates that both laevis and mulleri expression is stable. The appearance of paternal mitochondrial MDH does not correspond to the time when other mitochondrial components begin to increase in Xenopus. Multiple bands of MDH in both species and of oxidase in laevis are probably not due to the aggregation of subunits produced by different alleles at the same locus. There is no evidence for the formation of “hybrid” molecules consisting of subunits of both species.     

Multilocus isozyme systems in African lungfish,Protopterus annectens: distribution,differential expression and variation in dipnoans

Distribution of ADH, ALP, FBALD, GAPDH, G3PDH, G6PDH, GPI, LDH, MDH, PGM, and SOD was identified in retina, heart, muscle, liver, kidney, gills, brain, gut, lung and ovary of the African lungfish. Data are compared with patterns previously described in dipnoans and other vertebrates. The number of loci expressed for all enzymes was found to be similar to those of diploid Actinopterygii. Differences in the number of loci expressed in Amphibia were found for ALP, sG3PDH, GPI, LDH, MDH and SOD. Differences in tissue distribution were noted in ALP due to the absence of an intestinal-specific form typical of teleostean fish, amphibians, reptiles and birds, and in GPI and MDH, due to the tissue expression, as in primitive fish. There were also differences in LDH, where a third locus (LDH-C*) was expressed in the gills of Protopterus annectens and not in the retina or liver tissues, as in teleosts. LDH-A₄ was most common in all the tissues. Major differences were noted in the tissue patterns of protein expression in the three dipnoans compared. As expected, the least divergence was found between the two species belonging to the same family (Lepidosirenidae). The highest index of divergence was observed between Neoceratodus forsteri and Lepidosiren paradoxa, belonging to the families Ceratontidae and Lepidosirenidae, respectively. The divergence is revealed by changes at the enzyme and morphological levels. These results suggest that P. annectens occupies an interesting systematic position, its biochemical characteristics distinguishing it from N. forsteri, L. paradoxa, the advanced fish and amphibians.     

Diel activity patterns of the fish community in a temperate stream

The diel activity patterns of fishes in a temperate New Brunswick stream were studied during the summer over 5 years. Young‐of‐the year Atlantic salmon Salmo salar and blacknose dace Rhinichthys atratulus were more active during the day than at night, whereas lake chub Couesius plumbeus, brook trout Salvelinus fontinalis and adult white suckers Catostomus commersonii were more active at night than during the day. Because fishes were as likely to be nocturnal as diurnal, the data suggest that more night‐time sampling is needed to provide an unbiased view of fish community structure in temperate streams.     

Comparison of glycolytic and associated enzyme activities in the claw and tail muscles of the lobster with those of the mantle of Mytilus edulis

In the mussel Mytilus edulis, the activities of phosphoenolpyruvate car?ykinase (PEP-CK) and of malate dehydrogenase (MDH) are greater than those of pyruvate kinase (PK) and lactate dehydrogenase (LDH). The activities of PEP-CK are very low in the lobster (H. vulgaris.) and the ratio of MDH/LDH activities are 0·041 and 1·83 in the tail and the claw muscles respectively. Intracellular L-lactate and L-alanine concentrations suggest a different carbohydrate utilization in the tail and the claw muscle of the lobster. Consistent with this finding is the fact that L-lactate concentration is higher in the tail muscle than in the claw muscle; the opposite is true for L-alanine concentrations.     

Feeding ecology of three predacious aquatic insects and two fish in a riffle of the Speed River,Ontario

The food habits of two fish (Etheostoma flabellare and Rhinichthys atratulus) and three predacious aquatic insects (Nigronia serricornis, Paragnetina media and Agnetina capitata) were examined for one year in a riffle of the Speed River, Ontario. Adult E. flabellare fed primarily on net-spinning caddisflies whereas juvenile E. flabellare relied heavily on chironomids. Rhinichthys atratulus had a more varied diet composed of chironomids, Antocha larvae (Tipulidae) and net-spinning caddisflies. Two of the aquatic insect predators consumed large numbers of net-spinning caddisflies; however, P. media contained large percentages of Hydropsyche and Cheumatopsyche whereas N. serricornis fed primarily on non-hydropsychid net-spinners. The diet of A. capitata was composed largely of chironomids and mayflies. Schoener's diet overlap index () suggested that large E. flabellare and P. media had similar diets; however, E. flabellare ingested much larger prey than P. media. Adult E. flabellare contained a significantly greater proportion of larger prey items in comparison with benthic samples whereas similar comparisons with other predators showed no selection for larger prey. These data demonstrate that E. flabellare was size selective given the range of prey sizes available whereas the other predators were not. Also, there was little dietary overlap among the fish and invertebrate predators suggesting a partitioning of resources.     

Low-salinity stress in the American lobster,Homarus americanus,after chronic sublethal exposure to cadmium: Biochemical effects

Lobsters(Homarus americanus) were exposed to cadmium (6μg l⁻¹, 30 days) in flowing seawater, then held for 7 days in aerated “clean ” seawater at either ambient (27 ‰) or low (17 ‰) salinity. Cadmium exposure alone (ambient salinity) induced a general elevation of enzyme activity (heart, antennal gland, and muscle MDH; heart LDH and GPI), despite the probability of some clearance of cadmium from body tissues during the “clean ” seawater holding period. Low-salinity alone (non-exposed lobsters) caused a decrease of enzyme activity (AAT, LDH, GPI, PK) in most tissues examined, except for tail muscle IDH, the activity of which was increased, and MDH, which was significantly elevated above ambient controls in all tissues except heart. Most low-salinity effects were observed in tail muscle, and most cadmium effects, in heart; low-salinity effects outnumbered cadmium stress by nine to four. In heart and tail muscle of cadmium-exposed lobsters held at low salinity, each of the two stresses apparently operated to nullify the other's effects. The most prominent single biochemical response to these sublethal stresses was the elevation of MDH activity. The ratio MDH: LDH gave the clearest indication of overall relative stress.     

High-Altitude Adaptation of Yak Based on Genetic Variants and Activity of Lactate Dehydrogenase-1

This study investigates the molecular mechanism by which yaks (Bos grunniens) adapt to hypoxia based on lactate dehydrogenase (LDH). Three LDH1 variants of the yak were revealed in tissue extracts by electrophoresis, including LDH1-F, LDH1-M, and LDH1-S. Kinetic analysis using purified LDH1 variants showed that the yak LDH1-M variant exhibited a similar K _m (NADH) and the same mobility on a gel as bovine LDH1, and the LDH1-F variant showed significant differences in K _m values for NADH or pyruvate from the other two variants of yak LDH1 and bovine LDH1. Among the three muscles assayed, yak longissimus dorsi showed the highest LDH activity and the lowest malate dehydrogenase (MDH) activity; heart muscle was exactly the opposite. Our results suggest that the three LDH1 variants might play an important role in the adaptation to hypoxia.     

Expression of lactate dehydrogenase phenotypes in intraspecific and interspecific matings of two species of Xenopus

Starch gel electrophoresis has been used to examine lactate dehydrogenase phenotypes in two species of Xenopus and their hybrids obtained from reciprocal crosses. The patterns are complex, consisting of as many as 18 bands in some material. Differences between laevis and mulleri isozymes allow an evaluation of the contribution of both parents to the phenotypes of their hybrid offspring, and the determination of approximate times of paternal allele expression. The phenotype of early embryos resembles that of the maternal parent until hatching, when evidence of paternal influence is first apparent. Regardless of the early appearance of paternal enzyme, reciprocal hybrids bear a stronger resemblance to the maternal parent until well after tadpole growth begins. Once this maternal effect disappears, both laevis and mulleri appear to contribute to the LDH phenotype without predominance of the isozymes of either species. Evidence for the possible formation of “hybrid” enzymes consisting of subunits of both species in one active enzyme molecule is presented. Expression of LDH phenotype is variable in the unfertilized eggs of fertile hybrid females.     

Carbohydrate energy metabolism in Fasciola gigantica (trematoda)

Umezurike G. M. and Anya A. O. 1980. Carbohydrate energy metabolism in Fasciola gigantica (Trematoda). International Journal for Parasitology10: 175–180. Adult Fasciola gigantica contained 4.49 ± 0.06 % (mean ± S.D.) wet weight glycogen. Tissue homogenates contained high levels of malate dehydrogenase (MDH), NAD-linked malic enzyme (ME), Phosphoenolpyruvate carboxykinase (PEPCK) and lactate dehydrogenase (LDH). MDH, PEPCK and ME activities appeared to be localized in both cytosolic and mitoehondrial fractions, fumarase activity appeared to be predominantly mitochondrial whereas LDH and pyruvate kinase activities were cytosolic in distribution. Polyacrylamide gel electrophoresis revealed the predominance of LDH-1, LDH-2 and LDH-3 but only traces of LDH-4 and LDH-5 isoenzymes in the crude cytosolic fraction. LDH activity in the crude sample was inhibited by excess substrate (pyruvate). The mitoehondrial system showed NADH -cytochrome c oxidoreductase, succinate-cytochrome c oxidoreductase, NADH oxidase and some cytochrome c-oxygen oxidoreductase activities. Under anaerobic conditions, NADH-fumarate oxidoreductase and succinate-NAD + oxidoreductase activities of mitoehondrial preparations were stimulated in the presence of ADP and ATP respectively. Isolated mitochondria contained rhodoquinone and no ubiquinone, and isolated rhodoquinone was readily reduced by succinate in the presence of submitochondrial particles. Hydrogen peroxide was produced by submitochondrial particles in the presence or absence of KCN or in the presence of fumarate.     

Electrophoretic patterns of esterases and lactate-and l-malate-dehydrogenases from Flavobacterium species

Esterases and lactate-(LDH) and l-malate-(MDH) dehydrogenases of 64 strains of 13 species of Flavobacterium (particularly 46 strains of F. meningosepticum belonging to 15 different serotypes) were analyzed by horizontal polyacrylamide-agarose gel electrophoresis. Twenty-five esterase types were detected; these enzymes were distinguished by their spectra of hydrolytic activity towards five synthetic substrates (hydrolytic type) and their electrolytic mobilities (electrophoretic type). No variant of LDH was detected in all the strains of Flavobacterium. No variant of MDH was detected in three species: F. aquatile, F. branchiophilum, and F. breve and in serotype I, J and L. of F. meningosepticum. These results within the genus Flavobacterium permit precise identification, which may be useful for characterization of strains kept in collections as well as for epidemiological studies.     

Winter biology of the blacknose dace,Rhinichthys atratulus,in a southern Ontario stream

Synopsis The winter habitat of stream-dwelling blacknose dace Rhinichthys atratulus is identified and described from underwater observations in a southern Ontario stream. In late November as water temperatures declined below 5.0°C, dace moved into crevices beneath rubble with a mean maximum diameter of 15.9 cm. Focal point water depth was significantly greater in winter and mid-depth water velocity was significantly faster than measured in the summer. Dace were not observed in open water again until late March when water temperatures increased to 4.0°C. Winter diet analyses which showed minimal feeding were consistent with the seasonal changes in condition factor and percentage body water suggesting a general depletion of body reserves over winter. This physiological change was more pronounced in juveniles.     

Malate dehydrogenase isozymes in five species of sarcophagid flies (Sarcophagidae: Diptera)

Malate dehydrogenase banding patterns were analyzed by polyacrylamide gel electrophoresis in Sarcophaga ruficornis, S. argyrostoma, S. dux, S. invaria and S. peregrina. Two distinct zones of MDH activity, attributed to be the product of two gene loci, namely, MDH-1 and MDH-2 were observed. The MDH-1 locus revealed polymorphism for three electrophoretic phenotypes which are governed by two electromorphs MDH-1^a and MDH-1^b. The MDH banding pattern does not reveal any notable interspecific difference.