Some ecological relationships of larval ascaridoids from south-eastern Queensland marine fishes

Nine larval types within the genera Anisakis, Terranova, Thynnascaris and Contracaecum were recovered from south-eastern Queensland marine fishes. Data on (i) incidence, (ii) intensity of infection, (iii) host diet and (iv) habitat for each type suggest Anisakis is an open water type, Contracaecum an inshore, shallow water type and both Terranova and Thynnascaris have intermediate distributions. Host diets indicate Anisakis and Terranova are found in predators of nekton, Contracaecum and Thynnascaris in benthic feeders.     

In vitro activation of phosphoglucomutase by fructose 2,6-bisphosphate

The hexose bisphosphate activation of phosphoglucomutase was investigated with both plant (pea and mung bean) and animal (rabbit muscle) sources of the enzyme. Plant phosphoglucomutase was purified about 50-fold from seeds, and to a lesser extent, from seedlings of Pisum sativum L. cv Grenadier and seedlings of Phaseolus aureus. It was found that the plant enzyme was isolated in a mostly dephosphorylated form while commercial rabbit muscle phosphoglucomutase was predominantly in the phosphorylated form. Activation studies were done using the dephosphorylated enzymes. The range of activation constant (K_a) values were obtained for each bisphosphate were: for glucose 1-6-P₂, 0.5 to 1.8; fructose 2,6-P₂, 6 to 11.7; and fructose 1,6-P₂, 7 micromolar, respectively. Fructose 2,6-P₂ is known to occur in both plant and animal tissues at changing levels encompassing the K_a values found in this study; hence, these results implicate fructose 2,6-P₂ as a natural activator of phosphoglucomutase, particularly in plants. Also, glucose 1,6-P₂ has not been found in plants, and the method for measuring glucose 1,6-P₂ by monitoring the activation of phosphoglucomutase is not specific.     

The dissociation of aggregate forms of dextransucrase

Aggregate forms of dextransucrase obtained from Streptococcus sanguis dissociate in the presence of sodium dodecyl sulfate. However, the enzyme was unstable under these conditions. Nonionic detergents such as Triton X-100 stabilize the enzyme (A. W. Miller and J. R. Robyt, (1981) Fed. Proc. Fed. Amer. Soc. Exp. Biol.40, 1656), but do not cause disaggregation. The combination of both detergents, at concentrations below their critical micellar concentrations, is effective in dissociating and in stabilizing the enzyme. Polyacrylamide gel electrophoresis of the enzyme in the presence of the mixed detergents produced five major active enzyme forms, and two minor ones. The molecular weights of these forms were established by a modification of the procedure of Hedrick and Smith ((1968) Arch. Biochem. Biophys.113, 675–683).     

Genetic Variation in Some Populations of the Golden-Striped Salamander, Chioglossa lusitanica (Amphibia: Urodela), in Portugal

Genetic variation in the golden-striped salamander (Chioglossa lusitanica) was assessed in 231 individuals from four Portuguese populations by means of horizontal starch gel electrophoresis and isoelectric focusing. Three of 19 enzyme systems, representing 21 presumptive loci, were found to be polymorphic: phosphoglucomutase 1 (PGM1), peptidase B (PEPB), and peptidase D (PEPD). The observed average heterozygosity in Chioglossa lusitanica (0.027) is significantly lower than that observed for other amphibians, either urodeles or salamandrids. Differences in allele frequencies and the presence of private alleles are indicative of a high degree of population differentiation. PEPD, in particular, seems to be a diagnostic locus separating the southernmost population studied from the others.     

Differentiation of taeniid cestodes by enzyme electrophoresis

Le Riche P.D. and Sewell M.M.H. 1978. Differentiation of taeniid cestodes by enzyme electrophoresis. International Journal for Parasitology8: 479–483. Interspecific differences were shown by enzyme electrophoresis in thin starch gel between Echinococcus granulosus (from horses), Taenia hydatigena, T. multiceps, T. ovis, T. pisiformis, T. saginata, T. solium and T. taeniaeformis, Differences were seen with adenylate kinase, glucose phosphate isomerase, glutamate dehydrogenase, hexokinase and malate dehydrogenase. Multiple molecular forms of glucose phosphate isomerase were clear and most species could be differentiated by this enzyme alone.No differences attributable to anthelmintic effect were seen and zymogram patterns were identical for adult and cystic forms of each species. No differences were seen between specimens collected from various geographical locations.     

Allozyme variation in the fruit flies Dacus tryoni and Dacus neohumeralis (Tephritidae)

Electrophoretic variation in four specific proteins, an alcohol dehydrogenase, and an octanol dehydrogenase from adult body homogenates, an esterase from adult heads, and a tyrosinase from larval hemolymph, is described for laboratory populations of two sibling species, Dacus tryoni and Dacus neohumeralis. For each enzyme, a number of test crosses between different electrophoretic forms provided evidence that the observed variation was due to segregation of alleles at one structural gene locus.     

Isolation of a detergent-solubilized maltase/glucoamylase from rat intestine and its comparison with a maltase/glucoamylase solubilized by papain

Maltase/glucoamylase from the rat intestinal brush-border membrane was solubilized by homogenization of the intestinal mucosa in buffer containing 0.5% Triton X-100. After removal of the detergent with butan-1-ol, the enzyme was purified by chromatography on Sepharose 4B and DEAE-cellulose. The final specific activity was 70.3 units/mg of protein in six preparations, comparing favourably with the specific activity of 65.0 units/mg of protein of a pure papain-solubilized maltase/glucoamylase previously isolated and characterized by us [Flanagan & Forstner (1978) Biochem. J. 173, 553–563]. The two enzymes were compared. Both migrated as single bands with the same mobility on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, were eluted at the same volume from Sepharose 4B, and had the same sedimentation pattern in mannitol gradients. The amino acid composition was similar; content of total apolar residues differed by 1.0mol%. Antibodies prepared against either enzyme gave identical precipitin lines with each. Neither enzyme bound tritiated Triton X-100. The only difference noted was the tendency of the detergent-solubilized enzyme to aggregate on storage, whereas the papain-solubilized enzyme remained unchanged. Both enzymes had two N-termini, glycine and arginine. When the two enzymes were dissociated by boiling in sodium dodecyl sulphate, each exhibited the same five species on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Single N-termini were found in the two smaller species, 1 (glycine) and 2 (arginine), whereas larger species (3–5) had both N-terminal amino acids. Both the Triton- and papain-solubilized enzymes appear to be oligomers of species 1 and 2, indicating that the native enzyme contains two subunit types. Aggregation in aqueous solutions does not depend on a proteolytically susceptible peptide fragment at the N-terminus of either subunit.     

Genetic control of phosphoglucomutase variants in Saccharomyces cerevisiae

The three electrophoretic variants of phosphoglucomutase in Saccharomyces cerevisiae breeding stocks are produced by two unlinked genes, pgm-1 and pgm-2; pgm-1 contains two known alleles, pgm-1a and pgm-1b, each of which specifies a minor phosphoglucomutase component, and pgm-2 specifies the major phosphoglucomutase component.     

Some larval ascaridoids from south-eastern queensland marine fishes

A survey of 123 species of fishes from southeastern Queensland during 1975 revealed in 47 species nine distinct larval types of ascaridoid nematodes: Anisakis Type I, Terranova Types I and II, Contracaecum Types I and II, and Thynnascaris Types I, II, III and IV. These larvae are described and figured.     

Developmental regulation of juvenile hormone esterase in Trichoplusia ni: Its multiple electrophoretic forms occur during each larval ecdysis

Juvenile hormone esterase activity has been found during the intramoult period of each larval stadium in Trichoplusia ni. The activity is indistinguishable from that occurring during the final larval stadium, on the basis of its four isoelectric forms and kinetic data. The 4th- to 5th-instar intramoult peak in activity also occurs in other Lepidoptera (Heliothis virescens, Spodoptera exigua, Manduca sexta and Hyphantria cunea). Further, the final species also possessed a peak of activity during the intramoult period to the penultimate larval instar. The findings have important implications for the current concept that the function of juvenile hormone esterase is, by reason of its anti-juvenile hormone action, an enzyme of the last larval instar which enables metamorphosis to begin.     

Dynamics of a parasite assemblage of the Vermilion Rockfish Sebastes miniatus from northwestern Baja California,México

A parasite assemblage of Sebastes miniatus from northwestern Baja California, México, was composed of a total of 12 species: five ectoparasites (two monogeneans and three parasitic copepods) and seven endoparasites (two digeneans, one cestode, three nematodes, and one acanthocephala). Five of these parasites constituted new genera records to the genus Sebastes, and nine were new geographic records. The most abundant species were the endoparasites Parabothriocephalus sagitticeps, Hysterothylacium sp., and Anisakis sp., and the specific richness ranged from 1 to 8 parasite species per host. The most important parasite species in terms of prevalence were Microcotyle sebastis (93 %) and Anisakis sp. (92 %). The mean abundance of parasites found in S. miniatus showed significant variations over the year, with maximum values (31.7 individuals/host) in August, and minimum (0.39 individuals/host) in February. P. sagitticeps showed the highest mean intensity of infection (190.4 parasites/host), followed by Anisakis sp. (127.2 parasites/host) and Hysterothylacium sp. (46.6 parasites/host). The presence of larval stages of the nematodes Anisakis, Pseudoterranova, and Hysterothylacium is particularly important due to their high abundance and prevalence and because they may represent a human health risk (anisakiasis). Rockfishes (family Scorpaenidae) of the genus Sebastes constitute one of the most important groundfish resources in the American and Mexican northern Pacific Ocean, both for recreational and for the commercial fisheries of California and Baja California. These rockfish species makes up a substantial part of the Mexican cuisine.     

Identification of life cycle stages of the nematode Echinocephalus overstreeti by allozyme electrophoresis

Data presented in this study highlight the potential of allozyme electrophoresis in providing unequivocal genetic evidence for the identification of life cycle stages, particularly where species have complex life cycles. Adults of the nematode Echinocephalus overstreeti parasitize the elasmobranch Heterodontus portusjacksoni. The putative larval form which is morphologically dissimilar is found in two species of marine molluscs, Chlamys bifrons and Pecten albus. Electrophoretic analysis indicated that the adult and larval forms shared alleles at all of the 34 enzyme loci established. Furthermore, there were no fixed allelic differences between larval forms from different mollusc species.     

Protein polymorphism in native goats from central Argentina

Under the name of Creole goats, individuals of various origins, morphological and productivity characters are grouped. To characterize the genetic pool of goats from different sources in central Argentina, protein polymorphism revealed by electrophoresis was analyzed. A total of 109 adult animals from Santa Marı́a, Colón and Ischilı́n Districts, Córdoba Province, were studied. The red cells lysates were subjected to starch gel electrophoresis and specific staining procedures to reveal the activity of different enzymes and hemoglobin. Separation of plasmatic proteins was carried out by electrophoresis in polyacrylamide gel and stained with Coomassie Blue. From a total of 14 loci analyzed, eight were polymorphic in at least one flock. Four codominant alleles were detected in the locus βHemoglobin (βHb); three alleles in malic enzyme (Me), glucose 6 phosphate dehydrogenase (G6pdh) and lactate dehydrogenase (Ldh) and two alleles in catalase (Cat), transferrin (Tf), esterase-2 (Es-2) and phosphoglucomutase (Pgm). In all the cases, the observed genotype frequencies were not significantly different from those expected from Hardy–Weinberg equilibrium. The proportion of polymorphic loci (P%) varied between 21.4% and 42.9% and mean heterozygosity (H), between 0.061 and 0.117. Alleles at loci Cat, Me and Es-2 showed significantly different frequencies according to the sample origin. The flock from Colón District presented the highest levels of polymorphism. This is the only stock which includes hybrids of varying grade between Creole stocks and male specimens from Saanen and Nubian breeds.     

A mannose phosphate isomerase polymorphism in the Isopod Asellus aquaticus

A mannose phosphate isomerase locus in Asellus aquaticus is highly polymorphic in two populations examined in Britain. Breeding studies indicate four classes of electrophoretic alleles based on mobility differences, a further null allele, and probable genetically determined variation in enzyme activity between individuals with the same electrophoretic alleles. No linkage with the polymorphic glucose phosphate isomerase and phosphoglucomutase loci was detectable.     

Purification of a beta-Amylase that Accumulates in Arabidopsis thaliana Mutants Defective in Starch Metabolism

Amylase activity is elevated 5- to 10-fold in leaves of several different Arabidopsis thaliana mutants defective in starch metabolism when they are grown under a 12-hour photoperiod. Activity is also increased when plants are grown under higher light intensity. It was previously determined that the elevated activity was an extrachloroplastic β-(exo)amylase. Due to the location of this enzyme outside the chloroplast, its function is not known. The enzyme was purified to homogeneity from leaves of both a starchless mutant deficient in plastid phosphoglucomutase and from the wild type using polyethylene glycol fractionation and cyclohexaamylose affinity chromatography. The molecular mass of the β-amylase from both sources was 55,000 daltons as determined by denaturing gel electrophoresis. Gel filtration studies indicated that the enzyme was a monomer. The specific activities of the purified protein from mutant and wild-type sources, their substrate specificities, and K_m for amylopectin were identical. Based on these results it was concluded that the mutant contained an increased level of β-amylase protein. Enzyme neutralization studies using a polyclonal antiserum raised to purified β-amylase showed that in each of two starchless mutants, one starch deficient mutant and one starch overproducing mutant, the elevated amylase activity was due to elevated β-amylase protein.     

Purification and characterization of homogeneous sunflower seed acid phosphatase

Acid phosphatase (EC 3.1.3.2) from sunflower seed was purified 1800-fold to homogeneity using both conventional and affinity chromatographic methods. The purified enzyme was a mixture of two enzyme forms distinguishable by polyacrylamide gel electrophoresis (PAGE). Gel exclusion chromatography, which did not distinguish between the two forms, gave an apparent M, of 103 000. Preparative PAGE permitted the separation of the two forms, and SDS-PAGE showed that they contained equivalent peptide subunits of apparent M, 56 000 and 52 000. Amino acid analysis indicated that both enzyme forms have similar amino acid compositions. Data on substrate specificity and pH dependence is presented. The kinetic constants for hydrolysis of p-nitrophenyl phosphate as catalysed by sunflower seed acid phosphatase were independent of pH in the range 3-5. The enzyme was competitively inhibited by inorganic phosphate and non-competitively inhibited by phosphomycin.     

Multiple forms of peroxidase from Narcissus pseudonarcissus

Multiple forms of peroxidase from Narcissus pseudonarcissus were identified and separated by polyacrylamide gel electrophoresis. The enzyme forms were found to be particulate but could be solubilized in buffers of high ionic strength and high pH. Bulbs at different stages (dormant, early growth, flowering and post-flowering) were investigated and both the number and distribution of peroxidase forms were found to differ. The major peroxidase form in dormant bulbs was purified and displayed a number of notable properties including a MW of at least 10⁵, a high isoelectric point and the apparent absence of a heme prosthetic group.     

Genetic differentiation between two types of dark chub,Zacco temmincki,in Japan

Two types of the dark chub,Zacco temmincki, collected from 10 river systems in Japan were genetically characterized at 27 protein coding loci using starch-gel electrophoresis. They were fixed for different alleles at 13 loci. No hybrid individuals were observed, even in specimens collected in stations where both types appear sympatrically, indicating that each type of the dark chub represents a distinct species.     

The fate of multiple doses of infective larvae of Nematodirus battus in 8-month-old lambs and their effect on intestinal enzyme activity

Mapes C.J. and Coop R.L. 1973. The fate of multiple doses of infective larvae of Nematodirus battus in 8-month-old lambs and their effect on intestinal enzyme activity. International Journal for Parasitology3: 363–370. The fate of five daily doses of 60,000 infective larvae of Nematodirus battus was studied in 8-month-old lambs. On 18, 26 and 34 days after the last larval dose 4.0, 3.3 and 1.0 per cent of the total infective dose was recovered. Approximately 50 per cent of the worms recovered on these days were fourth-stage larvae. It is suggested that L5 and adult stages were preferentially lost from the hosts and that male worms were developing at a faster rate than the females. The populations of N. battus were smaller, contained higher proportions of fourth-stage larvae and shorter L5 and adult worms than those developing from similar infective doses in 3-month-old lambs. A transient decrease in alkaline phosphatase and maltase levels was found in the mucosa of the small intestine and was compared with the marked and persistent changes in mucosal enzyme activities found with similar infective doses in 3-month-old hosts.