Leucine aminopeptidase in exsheathing fluid of North American and Australian Haemonchus contortus

Rogers W. P. and Brooks F. 1978. Leucine aminopeptidase in exsheathing fluid of north American and Australian Haemonchus contortus. International Journal for Parasitology8: 55–58. Juveniles of Haemonchus contortus from north America and Australia produced exsheathing fluid containing leucine aminopeptidase when stimulated in tetraborate-carbon dioxide medium. Exsheathment in this medium was inhibited by 1, 10-phenanthroline, 10^?3M, and this inhibition was largely reversed by Zn²⁺, 10^?3M. This supports the view that the enzyme is produced by the juveniles and that it is concerned in exsheathment.     

Leucine aminopeptidase and exsheathing activity in preparations from Haemonchus contortus

Rogers W.P. and Brooks F. 1978. Leucine aminopeptidase and exsheathing activity in preparations from Haemonchus contortus. International Journal for Parasitology 8: 449–452. Exsheathing activity relative to leucine aminopeptidase activity (LAP) was greater in exsheathing fluid of infective juveniles of Haemonchus contortus than extracts of homogenates of the same organism. In both preparations the biological and enzyme activities were precipitated with acetone 20 v/v and ammonium sulphate, 40% saturation. Broad peaks of exsheathing and LAP activities obtained by sucrose density-gradient centrifugation and on Sephadex G150 overlapped but the peak of biological activity was always found on the low mol. wt. side of the LAP peak. LAP in exsheathing fluid was separated into two sharp peaks in polyacrylamide gradient-pore electrophoresis. In four experiments the major peak gave a mol. wt. within the limits 345,000–354,500. A minor peak was obtained at 1,800,000. Exsheathing activity remained broadly distributed but fell mostly on the low mol. wt. side of the major LAP peak.It is concluded that LAP cannot be the sole agent involved in exsheathment a lipase may be necessary to expose the substrate attacked by LAP.     

Gene activity and the development of early parasitic stages of nematodes

Actinomycin-D, 8 × 10^?6m, blocked the action of the stimulus from the host which normally initiates the development of the infective stage of Haemonchus contortus to the first parasitic stage. However, the action of the stimulus in initiating exaheathment was not affected by the antibiotic.The significance of these results in relation to the activity of the gene set of the first parasitic stage and the control of exsheathment of the infective stage is discussed.     

Role of mermithid nematodes in biological control of mosquitoes

Mermithid nematodes have been reported from at least 63 species of mosquitoes from all over the world, but until recently they have received little attention. Such nematodes are prime candidates as biological control agents because they have adapted to the life cycle of the host; are host specific; produce high levels of parasitism; kill the hosts; are easily handled; have a high reproductive potential; are free swimming and can be disseminated easily in the infective stage; and can be used in an inundative or inoculative manner to control mosquitoes. Nevertheless, with the exception of one mermithid species, little is known about factors that influence parasitism in mosquitoes. Also, only one species of mermithid has been successfully mass cultured to date. The exception is the mermithid Reesimermis nielseni. With this mermithid, high levels of parasitism can be obtained in natural populations of mosquitoes at reasonable dosages; preparasitic nematodes can be applied easily by using many of the standard techniques used to apply insecticides; and R. nielseni can establish itself in many of the sites in which it is released. The same potentials exist for mermithids of other aquatic insects.     

Zinc as a co-factor for an enzyme involved in exsheathment of Haemonchus contortus

The activity of concentrated exsheathing fluid of Haemonchus contortus against isolated sheaths was not inhibited by ethylenediamine tetra-acetic acid (EDTA), 10^?2 M, even when the concentrations of Mg and Mn were < 4 × 10^?4 M and < 0·9 × 10^?6 M respectively. Purified or diluted solutions of exsheathing fluid, even in the presence of Mg²⁺, 10^?3 M, were inhibited. Leucine aminopeptidase (LAP) in exsheathing fluid was active even at concentrations of Mg < 1·3 × 10^?5M. Concentrated solutions were partially inhibited by EDTA, 10^?2 M, at low concentrations of Mg; inhibition was increased in diluted and purified preparations.1,10-phenanthroline (Ophen) strongly inhibited exsheathing activity (Zn < 1 × 10^?6 M). When Zn²⁺, 10^?3 M was added, the inhibition was abolished. The hydrolysis of l-leucinamide was greatly increased in the presence of Ophen, 10^?4 M; this effect was abolished by adding Zn²⁺, 10^?3 M.It is suggested that exsheathing fluid from at least some ‘strains’ of H. contortus contains a Zn metallo-enzyme, probably LAP, which is involved in the process of exsheathment.     

Egg Enzymes of the Ruminant Nematode Trichostrongylus colubriformis

Summary

Preparations from eggs of the ruminant nematode Trichostrongylus colubriformis were examined for the presence of selected enzymes. Chitinase (0.12 units/mg total protein), collagenase (0.56 units/mg total protein), lipase (0.16 units/mg total protein) and acetylcholinesterase (0.61 units/mg total protein) were detected. Hatching of nematode eggs may require several forms of enzymatic activity.     

Studies of the life-history and energetics of marine and brackish-water nematodes

Summary An energy budget was constructed for the marine nematode Monhystera disjuncta. Respiration was measured with a modified Cartesian diver technique, in which the nematodes were kept in agar inside the diver head. The relationship between respiration and body weight was: R =1.53 W^0.75. Body growth was exponential during the juvenile phase, with a growth rate equal to 0.61 d^-1. After maturation the growth rate fell to 0.17 d^-1. Food uptake was measured in experiments with radiolabeled bacteria. In one series of experiments the accumulation of radiolabel in the nematodes was followed. In a second series the decrease in labeling was followed when pre-labeled nematodes fed on unlabeled bacteria. A model for label uptake permitted the calculation of assimilation efficiency and consumption rates. Consumption rates thus measured, correspond well to those calculated from the growth, reproduction and respiration rates. Assimilation efficiency was low, around 25%. Production efficiency (P/(P+R)) was high: 60% for the population at stable age distribution, and up to 75% for reproducing females. This seems to be a general feature in nematodes.     

Studies of the life-history and energetics of marine and brackish-water nematodes

Summary Aspects of the demography of Monhystera disjuncta were investigated at different temperatures (in agnotobiotic cultures) and in different feeding conditions (monoxenic cultures with different bacterial strains, and different densities in the feeding suspension with one strain). Embryonic development time, minimum generation time, egg deposition rate and adult longevity depend on temperature, quality and quantity of food offered. Body mass at maturity is an allometric function of food density. It is shown that a previously inferred selectivity in food uptake is an artifact of culture conditions. pH buffering and addition of sterols permit culture of the species on a wide variety of bacterial strains. M. disjuncta is less well adapted to take advantage of high food density than are nematodes from polysaprobic environments. The animals channel surplus energy intake into a larger body mass, without being able to increase their rate of population growth accordingly.     

Bile and bile salts and exsheathment of the intestinal nematodes Trichostrongylus colubriformis and Nematodirus battus

1972. Bile and bile salts and exsheathment of the intestinal nematodes Trichostrongylus colubriformis and Nematodirus battus. International Journal for Parasitology, 2: 433–438. Exsheathment of T. colubriformis was potentiated at physiological levels of CO₂ by bile and bile salts. Lamb bile and crude sodium taurocholate preparations produced the greatest increase in exsheathment while rabbit bile, sodium glycocholate and deoxycholate had less pronounced effects. Exsheathment in bile and taurocholate occurred at a pH above 4 and it is therefore suggested that infective larvae which fail to exsheath in the abomasum could well do so in the proximal part of the small intestine. Pure sodium taurocholate was found to greatly potentiate exsheathment of N. battus in vitro but this occurred at a pH below 3 and thus the action of this bile salt could not affect exsheathment in the host.     

Diapause and prolonged development in the embryo and their ecological significance in two cicadas, Cryptotympana facialis and Graptopsaltria nigrofuscata

The seasonal timing mechanism of egg hatching was examined in two cicadas, Cryptotympana facialis and Graptopsaltria nigrofuscata, with different but overlapping geographical distributions. These species lay eggs in summer, and nymphs hatch in the summer of the following year after egg durations of 10-12 months. When eggs were maintained at 25 °C from oviposition, both the species entered embryonic diapause within 60 days irrespective of photoperiod, but at different developmental stages between the two species. The optimal temperature for diapause development was approximately 15 °C in both the species. The development rate for postdiapause morphogenesis increased linearly with temperature in the range of 20-27.5 °C in C. facialis, and of 15-25 °C in G. nigrofuscata. The lower development threshold and the sum of effective temperatures were computed as 14.3 °C and 715.3 day-degrees in C. facialis and 12.1 °C and 566.6 day-degrees in G. nigrofuscata, respectively. The hatching dates predicted by these large thermal constants accorded with the hatching dates observed in the field, i.e., late June and mid-July in G. nigrofuscata and C. facialis, respectively. Therefore, the high thermal requirements for postdiapause development compel the cicadas to hatch in summer.     

Geographical variation in species diversity: A comparison of marine polychaetes and nematodes

The aim of this study was to determine if marine species diversity was influenced by geographical location and whether it was higher at lower latitudes. Artificial collectors (made of nylon pan scourers) were employed as a standard substratum for the colonisation of marine invertebrates inhabiting subtidal (12 to 15 m) hard, rocky bottom substrata. These artificial substrate units (ASUs) were deployed at different latitudes including northern and southern temperate (South West England, UK and New Zealand), tropical (Trinidad and Tobago, West Indies) and polar (Signy Island, Antarctica) areas. The polychaetes, representative of the macrofauna and the nematodes, representative of the meiofauna fractions of the total invertebrate fauna collected were analysed.Neither polychaete nor nematode species diversity showed a trend based on latitude and each taxon showed a different pattern of diversity variation in relation to location. Polychaete diversity varied from area to area with highest species diversity occurring in the southern temperate (New Zealand). Nematode species diversity however was similar for the northern and southern temperate (UK and New Zealand) and the tropical area (Trinidad and Tobago). Thus, although the number of locations studied was limited, these data do not conform to a gradient in species diversity with latitude as has been previously supposed. The success of ASUs to compare species diversities in standardised habitat units augurs well for their future use in other ecological areas such as biogeographical or pollution studies.     

Free-living nematodes and macrobenthos in a high-latitude glacial fjord

Kongsfjord is an open glacial fjord on the west coast of Svalbard, where the influence of the West Spitzbergen current ameliorates the effects of high latitude (79°N). The fjord is heavily influenced by glacial discharges of meltwater, ice and till, and related environmental gradients in sediments from the glaciers to the open sea include sediment deposition, organic content and disturbance. Other factors, such as the formation and break up of sea ice, also affect benthic communities. In this study spatial patterns in nematode and macrofaunal communities, in samples collected using box-corers and van Veen grabs during a cruise in September 1997, are described, compared and contrasted. Non-parametric multivariate analyses demonstrate that there were clear differences in community structure between stations in both macrofaunal and nematode assemblages. At stations where macrofauna were sampled using both box-cores and grabs there were also significant differences between samples collected by different methods, although there is evidence that these were influenced in part by slight differences in sampling location. Some evidence of disturbance to macrofaunal assemblages in the centre of the fjord is apparent. Macrofaunal community composition varied most closely with a combination of depth and sediment C : N ratio, whereas that of nematodes varied most closely with C : N alone. Proportions of feeding groups of nematodes showed little variation along the fjord. There is no evidence of a specialised nematode assemblage inhabiting the part of the fjord subject to the heaviest deposition of sediment. The taxonomic distinctness of nematodes decreased with increasing distance from the source of disturbance. This is in contrast to studies showing that the taxonomic distinctness of nematodes tends to decrease with increasing anthropogenic stress.     

A demonstration of bacterial conjugation within the alimentary canal of Rhabditis nematodes

Abstract: Rhabditis nematodes fed a diet of Escherichia coli defecate viable undigested bacteria. These bacteria retain phenotypic characteristics, including those encoded on plasmids. Nematodes can survive a 2-min surface sterilization with 2% chlorine bleach; internalized bacteria also survive this treatment and are released in the nematode wastes. Bacteria alone or on the surface of dead nematodes are unable to survive incubation with this solution. There were 3.2 × 10⁵ viable bacteria per nematode, indicating that sufficient bacteria were present for gene transfer. Transconjugants ( lac ⁻ nal ^R str ^R cm ^R) were recovered in the nematode fecal material following a protocol where nematodes were initially fed a plasmidless lac ⁻ nal ^R str ^S cm ^S E. coli and then, after surface sterilization, a lac ⁺ nal ^S E. coli plasmid donor containing the conjugative R100JA ( str ^R cm ^R) plasmid. The presence of plasmids in the transconjugants was confirmed by gel electrophoresis. The occurrence of conjugation in the gut was confirmed by dissection of individual surface-sterilized nematodes and isolation of transconjugants.     

Carbonic acid as the host signal for the development of parasitic stages of nematodes

Petronijevic T., Rogers W. P. and Sommerville R. I. 1985. Carbonic acid as the host signal for the development of parasitic stages of nematodes. International Journal for Parasitology15: 661–667. This paper gives results on which may be based an identification of the component of the system CO₂ + H₂O ai H₂CO₃ ai H⁺ HCO₃^? which acts as the stimulus from the animal host for some nematodes. Using infective juveniles of Nematospiroides dubius and Haemonchus contortus, the effects on exsheathment of (1) low pCO₂ values, (2) the presence of carbonic anhydrase in the stimulating medium, and (3) the inhibition of carbonic anhydrase within the juveniles have been examined. The results lead to the suggestion that it is the “readily available” undissociated H₂CO₃, or H₂CO₃ + HCO₃^? which is the critical factor in the stimulus for development. The wide range of [H⁺]s over which “readily available” H₂CO₃ is present in physiological environments suggests that this host signal may be important for infection with many species.     

Secretion of mutant leucine-specific binding proteins with internal deletions in Escherichia coli.

The leucine-specific binding protein, encoded by the livK gene, is located in the periplasm of E. coli. The present study is an attempt to identify intragenic regions that determine the efficiency of its secretion into the periplasm. C-terminal deletions or fusions of the livK gene to trpA (encoding the alpha subunit of tryptophan synthetase) were secreted with little loss of efficiency [1]. A series of deletions was constructed at the unique Sphl site within livK, near the 5' end of the region coding for the mature protein. Between 16 and 113 amino acids were deleted in the amino-terminal one-third of the protein. A few of these deletions were located within a few amino acids of the signal sequence processing site. Deletions extending within thirteen residues of the processing site were processed and secreted more slowly than normal. Secondary structure predictions suggested that the alpha-helical core region of the signal sequence extends into the mature protein in the case of the slow processing mutants, perhaps interfering with the recognition site for leader peptidase or other secretory components. These results suggest that the conformation around the signal processing site may be a critical factor in determining the efficiency of secretion. During the course of this study, it was found that the difference in molecular weight between precursor and mature forms of some binding protein mutants, as judged by SDS-PAGE, was much greater than could be accounted for by processing of the signal sequence. This anomalous mobility on gels, however, could be eliminated by performing SDS-PAGE in the presence of 6 M urea.     

Naturally occurring seco-ring-A-triterpenoids and their possible biological significance

The literature appearing up to mid 1984 on the isolation and identification of naturally occurring seco-ring-A triterpenoids is reviewed. The distribution of these triterpenoids in the plant kingdom is discussed. Their biological action is considered in relation to plant protection. It is concluded that ring-A fission is a general process in the oxidative degradation of tetra- and pentacyclic triternoids.     

Electrodynamics of interactions in electrolyte solutions and their biological significance

Attention is drawn to the fact that the interaction of charges in aqueous solutions of electrolytes, such as media having physiological characteristics, depends not only on the distance between interacting charges but also on the frequency that determines their dynamics. This fact has significant consequences for some biological processes and their kinetics. The analysis of reasons for charge shielding, including the dynamic effects, shows that, even at distances exceeding the Debye length, electric interactions in systems similar to physiological are effective provided that charges move with frequencies higher than 250 MHz. For each electrolyte solution, the threshold frequency (Maxwell frequency) can be found, which determines the transition from the conducting to the dielectric mode of interactions of charges in physiological solutions.     

Glycoproteins of trypanosomes: their biosynthesis and biological significance

1. Trypanosomes are unicellular parasites that cause human sleeping sickness in Africa and Chagas' disease in South America. Glycoproteins are important components of their plasma membrane. 2. The bloodstream form of the extracellular salivarian African trypanosome (e.g. Trypanosoma brucei) has the ability to express on its cell surface a repertoire of variant surface glycoproteins (VSGs) and in so doing, evades the immune response of the host (antigenic variation). 3. The VSG is probably synthesized initially in a manner like that of the membrane-bound glycoproteins of mammalian systems, but it also undergoes some novel post-translational modifications. 4. The stercorarian South American trypanosome (Trypanosoma cruzi) is an intracellular parasite which expresses different glycoproteins on its plasma membrane at various stages of its life-cycle, but does not exhibit antigenic variation. 5. The biosynthesis and functions of trypanosomal glycoproteins are compared with those of mammalian glycoproteins, and are discussed with particular reference to potential targets for chemotherapy and immunotherapy of trypanosomiasis.     

Chromenes and benzofurans of the asteraceae,their chemistry and biological significance

Chromenes (benzopyrans) and benzofurans are characteristic natural products of certain tribes of the Asteraceae. This review highlights recent findings