Worm Tissues Research Articles

Sublethal toxicity and biotransformation of pyrene in Lumbriculus variegatus (Oligochaeta)

The aim of this work was to study the toxicity and biotransformation of polyaromatic hydrocarbon (PAH) pyrene in the oligochaete aquatic worm, Lumbriculus variegatus. PAHs are ubiquitous environmental pollutants that pose a hazard to aquatic organisms, and metabolizing capability is poorly known in the case of many invertebrate species. To study the toxicity and biotransformation of pyrene, the worm was exposed for 15 days to various concentrations of water-borne pyrene. The dorsal blood vessel pulse rate was used as a sublethal endpoint. Pyrene biotransformation by L. variegatus was studied and the critical body residues (CBR) were estimated for pyrene toxicity. The toxicokinetics of pyrene uptake was evaluated. A combination of radiolabeled (14C) and nonlabeled pyrene was used in the exposures, and liquid scintillation counting (LSC) and high-pressure liquid chromatography were employed in both water and tissue residue analyses. The results showed that L. variegatus was moderately able to metabolize pyrene to 1-hydroxypyrene (1-HP), thus demonstrating that the phase-I-like oxidizing enzyme system metabolizes pyrene in L. variegatus. The amount of the 1-HP was 1-2% of the amount of pyrene in the worm tissues. The exposure to pyrene reduced the blood vessel pulse rate significantly (p<0.05), showing that pyrene had a narcotic effect. The estimated CBRs remained constant during the exposure time, varying from 0.120 to 0.174 mmol pyrene/kg worm wet weight. The bioconcentration factors (BCF) decreased as exposure concentration increased. It was suggested that the increased toxicity of pyrene accounted for the decrease in BCFs by lowering the activity of the organism.

Earthworms as bioindicator of metals (Zn, Fe, Mn, Cu, Pb and Cd) in soils: Is metal bioaccumulation affected by their ecological category?

The importance of earthworms in metal pollution monitoring is widely recognized in terrestrial ecosystems. Metal bioaccumulation by soil-dwelling earthworms can be used as an ecological indicator of metal availability in soils. In this study, we quantify the level of DTPA extractable metals in casts and tissues of earthworms (endogeic: Metaphire posthuma (Vaillant) and anecic: Lampito mauritii Kinberg) and ingesting soils, collected form cultivated land, urban garden and sewage soils. Soil and worm casts collected from sewage and cultivated land showed the greater metal concentrations. The concentration of Zn, Fe, Pb and Mn in earthworm casts was in the order: sewage soil > cultivated land > urban garden, while for Cu and Cd the order was cultivated land > sewage soil > urban garden. Data suggested that the level of DTPA extractable metals was higher than that of surrounding soils. We got close relationships between metal concentration in worm tissues and surrounding soils: Zn ( r 2 = 0.94 and 0.89, P < 0.01 for both), Fe ( r 2 = 0.95 and 0.97, P < 0.01 for both), Cu ( r 2 = 0.93 and 0.96, P < 0.01), Pb (0.63, P < 0.01 and 0.57, P > 0.05), and Cd ( r 2 = 0.15, P > 0.01 and 0.75, P < 0.01), respectively, for M. posthuma and L. mauritii. The study clearly indicates that earthworms have efficient potentials for bioaccumulation of metals in their tissues which can be used as an ecological indicator of soil contaminations. A species-specific metal accumulation pattern was observed in studied earthworms. Comparatively, endogeic showed the higher metal contents in their tissues than anecic ( t-test: P < 0.05); collected form different habitats studied. Data suggested that species-specific feeding behaviour, earthworm niche structure, ecological category of inhabiting earthworm and even horizontal distribution of contaminants in soil layers are some major determinant for metal accumulation patterns in soil dwelling earthworms. The difference in burrowing patterns can influence the patterns of metal bioaccumulations between endogeic and anecic, although other factors are also contributory. Further more detailed study is still required to elaborate the proposed hypothesis.

A comparison of the protein constituents of the major body compartments of the dog roundworm, Toxocara canis

An analysis of the protein profiles of intact worms and isolated tissues of adult male and female Toxocara canis worms was conducted. Soluble proteins recovered from homogenized whole specimens and dissected tissues (body wall, reproductive tract, esophagus and intestine) of T. canis adults from several different canine hosts were separated by size using gradient sodium dodecyl sulfate electrophoresis (SDS-PAGE) and visualized with silver staining. SDS-PAGE profiles of worms from different hosts were found to be virtually identical irrespective of sex or tissue type. Recovered proteins ranged in size from 3.4 to 325 kDa. As expected, variations existed between the protein profiles of different body tissues, with only slight variations between the sexes. The largest number of recovered proteins was present in the female reproductive tract extracts.

Bioaccumulation of Perfluorochemicals in Sediments by the Aquatic Oligochaete Lumbriculus variegatus

Bioaccumulation of perfluoroalkyl sulfonates, perfluorocarboxylates, and 2-(N-ethylperfluorooctane sulfonamido) acetic acid (N-EtFOSAA) from laboratory-spiked and contaminated field sediments was assessed using the freshwater oligochaete, Lumbriculus variegatus. Semistatic batch experiments were conducted to monitor the biological uptake of these perfluorochemicals (PFCs) over 56 days. The elimination of PFCs was measured as the loss of PFCs in L. variegatus exposed to PFC-spiked sediment for 28 days and then transferred to clean sediment. The resultant data suggest that PFCs in sediments are readily bioavailable and that bioaccumulation from sediments does not continually increase with increasing perfluorocarbon chain length. Perfluorooctane sulfonate (PFOS) and perfluorononanoate were the most bioaccumulative PFCs, as measured by laboratory-based estimated steady-state biota sediment accumulation factors (BSAFs) and BSAFs measured using contaminated field sediments. Elimination rate constants for perfluoroalkyl sulfonates and perfluorocaroboxylates were generally smaller than those previously measured for other organic contaminants. Last, a PFOS precursor, N-EtFOSAA, accumulated in the worm tissues and appeared to undergo biotransformation to PFOS and other PFOS precursors. This suggests that N-EtFOSAA, which has been detected in sediments and sludge often at levels exceeding PFOS, may contribute to the bioaccumulation of PFOS in aquatic organisms.

Ultrastructure of a spermatid transport system in the mature proglottids of experimental Taenia crassiceps (WFU strain)

In classical textbooks of parasitology, the mature proglottids of taeniids are depicted as structures in which the individual testis are connected to the vas deferens through the vas efferens system, usually depicted as a network of channels. From our morphological analyses of proglottids in the cestode Taenia crassiceps, we have been unable to identify this channel network. It is unclear how the spermatids are transported from the testes to the vas deferens, as is unresolved the location of the cells responsible for the production of testosterone (Leydig cells) or the possible equivalent of Sertoli cells, necessary for the differentiation process of these cells. In this experimental work, we have examined the ultrastructure of tissues in the vicinity of the vas deferens in mature proglottids obtained from the intestines of hamsters infected with cysticerci from the peritoneum of infected mice. Worm tissues were fixed, processed, and sectioned for transmission electron microscopy. Significant areas of the testis epithelia emitted cytoplasmic projections surrounded by extracellular matrix, where they appear as septated pockets enclosing free axonemes and spermatids. Vas efferens walls are made up of nucleated cells with cytoplasm annealing to each other through cell membrane junctions. Lodged between the junctions are membrane-bound pouches with dense granules found as aggregates or aligned in a semicircular array. The efferens wall exhibits numerous spermatids emerging into the lumen, an observation that suggests the epithelial wall may have the maturing functions of Sertoli cells of vertebrates. Large cells adjacent to the vas efferens contained prominent rough endoplasmic reticulum and large mitochondria, characteristics described for Leydig cells of vertebrates. Our observations suggest that taeniid spermatids are either transported from the testes to the vas system by epithelial pockets or that the epithelial pockets may be cross-sections of a highly coiled vas efferens system.

A persistent mitochondrial deletion reduces fitness and sperm performance in heteroplasmic populations of C. elegans

BackgroundMitochondrial DNA (mtDNA) mutations are of increasing interest due to their involvement in aging, disease, fertility, and their role in the evolution of the mitochondrial genome. The presence of reactive oxygen species and the near lack of repair mechanisms cause mtDNA to mutate at a faster rate than nuclear DNA, and mtDNA deletions are not uncommon in the tissues of individuals, although germ-line mtDNA is largely lesion-free. Large-scale deletions in mtDNA may disrupt multiple genes, and curiously, some large-scale deletions persist over many generations in a heteroplasmic state. Here we examine the phenotypic effects of one such deletion, uaDf5, in Caenorhabditis elegans (C. elegans). Our study investigates the phenotypic effects of this 3 kbp deletion.ResultsThe proportion of uaDf5 chromosomes in worms was highly heritable, although uaDf5 content varied from worm to worm and within tissues of individual worms. We also found an impact of the uaDf5 deletion on metabolism. The deletion significantly reduced egg laying rate, defecation rate, and lifespan. Examination of sperm bearing the uaDf5 deletion revealed that sperm crawled more slowly, both in vitro and in vivo.ConclusionWorms harboring uaDf5 are at a selective disadvantage compared to worms with wild-type mtDNA. These effects should lead to the rapid extinction of the deleted chromosome, but it persists indefinitely. We discuss both the implications of this phenomenon and the possible causes of a shortened lifespan for uaDf5 mutant worms.

Heavy Metal Concentrations in Water, Sediments and Body Tissues of Red Worm (Tubifex spp.) Collected from Natural Habitats in Mumbai, India

Live feeds, especially Tubifex spp., which are collected from a wide variety of polluted habitats, are used by aquarium fish keepers in India. These habitats receive domestic sewage and industrial wastes from nearby residential and industrial areas. Reports of morbidity and mortality from aquarium fish culturists in and around Mumbai led to the present investigations on the ecology of these habitats with a view to assess the water quality, presence of heavy metals in the environment and their bioaccumulation in Tubifex worms, and to examine whether these habitats could be exploited to meet the demand of the industry. Six natural red worm (Tubifex spp.) collection centres in Mumbai and Thane districts of Maharashtra state in India constituting a major source of live Tubifex supply to aquarium fish industry were evaluated for pollution, heavy metal concentration in water, sediments and in the body tissues of Tubifex. Data revealed the presence of heavy metals in water and sediments at collection sites and bioaccumulation of cadmium, iron, lead, zinc and copper in body tissues of Tubifex worms. Cadmium ranged from 2.38 to 7.21 mg/kg, iron 671.9 to 5738 mg/kg, lead 14.95 to 33.49 mg/kg, zinc 60.20 to 166.60 mg/kg and copper 29.38 to 108.90 mg/kg of dry Tubifex worms. The study suggests that all the six collection sites are polluted and the red worms contaminated with heavy metals and hence, unfit for use in aquaria or feeding any variety of fish or crustaceans in the hatcheries.

A balancing act between the X chromosome and the autosomes

Dosage compensation equalizes gene dosage between males and females, but its role in balancing expression between the X chromosome and the autosomes may be far more important. Now, DNA microarrays have shown equality between the average expression of X-linked genes and that of autosomal genes, in male and female tissues of flies, worms and mice.

Antibiotic Chemotherapy of Onchocerciasis: In a Bovine Model, Killing of Adult Parasites Requires a Sustained Depletion of Endosymbiotic Bacteria (WolbachiaSpecies)

Development of a drug lethal to adult Onchocerca volvulus (i.e., macrofilaricide) is a research priority for the control of human onchocerciasis. Using bovine O. ochengi infections, we investigated the effects of oxytetracycline administered in a short intensive regimen (SIR; 10 mg/kg daily for 14 days), compared with a prolonged intermittent regimen (PIR; 20 mg/kg monthly for 6 months) or a combination of both (COM), on the viability of adult worms and their endosymbiotic bacteria (Wolbachia species). The long-term treatments eliminated >80% (COM) or >60% (PIR) of adult female worms (P<.001), and the COM regimen effected a sustained depletion of Wolbachia organisms. Conversely, SIR was not macrofilaricidal and only transiently depleted Wolbachia densities, which repopulated worm tissues by 24 weeks after treatment. These results unequivocally demonstrate the macrofilaricidal potential of tetracyclines against Onchocerca infection and suggest that intermittent, protracted administration will be more effective than continuous shorter term treatment.

Impact of heavy metals on enzymatic activity of substrate and on composting worms Eisenia fetida

The effect of heavy metals Cu and Zn on dehydrogenase and protease activity of the substrate during vermicomposting was investigated. Three dosages of Cu and Zn were tested in mesocosm experiments to investigate their bioaccumulation and impact on the reproduction of Eisenia fetida. Cu accumulated within the worm tissues in dosage concentrations up to a maximum level of 213 mg Cu kg −1. The number of juveniles decreased from the lowest to highest dosages of Cu and Zn after 10 weeks of the experiment. Dehydrogenase showed a strong negative correlation ( P < 0.01) with increased dosage of Cu, while protease remained unaffected. An overall reduction on dehydrogenase activity with increasing dosages of Cu and Zn indicated that these metals would impact detrimentally on the soil microbiology and consequently the stabilisation of the dosed media.

Effects, transfer, and fate of RDX from aged soil in plants and worms

The objectives of this study were to provide data that can be used to predict exposure-based effects of RDX in aged soil on multiple endpoint organisms representing two trophic levels. These data can be used for defining criteria or reference values for environmental management and conducting specific risk assessment. Dose–response experiments formed the basis for the evaluation of toxic effects and transfer of contaminants from soil into two trophic levels. Long-term exposure tests were conducted to evaluate chronic, sublethal, toxicity and transfer of aged soil-based explosives, with RDX as main contaminant. In these tests, plants were exposed for 55 days in the greenhouse, biomass was determined and residues of explosives parent compounds and RDX metabolites were analyzed using HPLC techniques. Worms were exposed for 28 days ( Eisenia fetida) and 42 days ( Enchytraeus crypticus) in the laboratory, biomass and number were determined, and tissues were analyzed for explosives compounds. The plants tolerated concentrations up to 1540 mg RDX kg −1 soil-DW. Biomass of Lolium perenne was not significantly related to soil-RDX concentration, while biomass of Medicago sativa significantly increased. No screening benchmark for RDX in soil for plants was calculated, since concentrations up to 1540 mg kg −1 soil failed to reduce biomass by 20% as required for a LOEC. RDX, RDX-metabolite MNX, and accompanying HMX concentrations in plants were significantly related to concentrations in soil after 55 days of exposure (RDX: R 2 = 0.77–0.89; MNX R 2 = 0.53–0.77; HMX: R 2 = 0.67–0.71). The average bioconcentration factors (BCF) were for RDX 17 in L. perenne and 37 in M. sativa, and for HMX 2 in L. perenne and 44 in M. sativa. The worms also tolerated concentrations up to 1540 mg RDX kg −1 soil-DW. Biomass of E. fetida adults decreased with soil-RDX concentration, and a LOEC of 1253 mg kg −1 soil-DW was estimated. RDX concentrations in E. fetida were significantly related to concentrations in soil after 28-day exposure ( R 2 = 0.88). The average BCF in E. fetida for RDX was 1. Because in response to exposure to RDX-contaminated soil the RDX concentrations in plants increased initially and decreased subsequently, while those in worms increased continuously, RDX in worm tissues may accumulate to higher concentrations than in plant tissues, regardless of the low average BCF for worms.

Effects of gamma radiation on Brugia malayi infective larvae and their intracellular Wolbachia bacteria

Prior studies have shown that irradiated filarial larvae are developmentally stunted but capable of inducing partial immunity to filariasis in animals. The mechanisms for these effects are poorly understood. Recent studies suggest that intracellular Wolbachia bacteria are necessary for the normal development, reproduction and survival of filarial nematodes. The purpose of this study was to examine the effects of irradiation on Wolbachia in Brugia malayi infective larvae (L3) and on L3 development. The L3 were exposed to 0, 25, 35, 45, 55, 65 or 75 krad of gamma irradiation from a (137) Cesium source and cultured in vitro at 37 degrees C in NCTC/IMDM medium with 10% FCS for 12 days. Irradiation prevented molting of L3 to the L4 stage in a dose-dependent manner. Electron microscopy studies showed that irradiation damaged Wolbachia (25 krad) or cleared them from worm tissues (45 krad). In addition, majority of the irradiated L3s failed to develop the L4 cuticle. Real-time PCR studies showed that irradiation reduced Wolbachia DNA in worm tissues. Parallel in vivo studies confirmed decreased development of irradiated L3 in jirds, with associated effects on Wolbachia. Jirds injected s.c with normal L3 developed antibodies to Wolbachia surface protein (wsp) shortly after the onset of microfilarial patency. In contrast, jirds injected with irradiated L3 did not develop microfilaremia or antibodies to wsp. Additional studies are needed to test the hypothesis that irradiation retards growth and development of filarial L3 by killing Wolbachia.

Brugia malayi: effects of gamma radiation on adult worms and their intracellular Wolbachia bacteria

Prior studies have shown that intracellular Wolbachia endobacteria are necessary for the normal development, reproduction, and survival of filarial nematodes. The purpose of this study was to examine effects of gamma radiation on Wolbachia and reproduction in Brugia malayi adult worms. Worms were exposed to 0, 10, 25, 45, 75, and 105 krad of gamma radiation from a 137cesium source and cultured in vitro for 10 days. Irradiation reduced production of microfilariae in a dose-dependent manner. Embryograms of irradiated female worms showed dose-related abnormalities with arrested development at the early embryo stage. Irradiation reduced the viability of adult worms in a dose-dependent manner, but no lethal effect was observed. Electron microscopy studies showed that irradiation cleared Wolbachia from worm tissues. Real-time polymerase chain reaction studies demonstrated greatly reduced Wolbachia DNA in irradiated worms. These effects are essentially the same as those observed in adult worms treated with doxycycline. These studies suggest that effects of irradiation on reproduction in Brugia malayi may be caused by effects of irradiation on Wolbachia.

A light and electron microscopic study on the synergistic effect of pyrantel and the febantel metabolite febendazole on adult Toxocara canis in vitro.

In the present study, we investigated the in vitro effects on the motility and morphology of tissues and organs of Toxocara canis of the two drug components of Drontal Plus and Welpan, pyrantel and fenbendazole (the active metabolite of the prodrug febantel), both alone and in combination. Although there was no significant difference observable between the effects of the single drugs and the drug combination on worm motility, the synergistic effect of pyrantel and fenbendazole was manifested by morphological alterations seen by light and electron microscopy. Thus, an earlier onset of damage to worm tissues and organs could be observed compared to the application of the individual drugs. In addition, a higher degree of damage and an increased number of vital organs were involved. There was dramatic, significantly greater and irreversible damage to the hypodermis, longitudinal muscle, intestine, nerve cords and genital organs induced by the pyrantel/fenbendazole combination. We hypothesise that these synergistic effects will also take place when dogs are treated either with Drontal Plus or Welpan in which lower dosages will be sufficient to destroy the worms.

Lethal critical body residues as measures of Cd, Pb, and Zn bioavailability and toxicity in the earthwormEisenia fetida

Background. Earthworm heavy metal concentrations (critical body residues, CBRs) may be the most relevant measures of heavy metal bioavailability in soils and may be linkable to toxic effects in order to better assess soil ecotoxicity. However, as earthworms possess physiological mechanisms to secrete and/or sequester absorbed metals as toxicologically inactive forms, total earthworm metal concentrations may not relate well with toxicity. The objectives of this research were to: i) develop LD50s (total earthworm metal concentration associated with 50% mortality) for Cd, Pb, and Zn; ii) evaluate the LD50 for Zn in a lethal Zn-smelter soil; iii) evaluate the lethal mixture toxicity of Cd, Pb, and Zn using earthworm metal concentrations and the toxic unit (TU) approach; and iv) evaluate total and fractionated earthworm concentrations as indicators of sublethal exposure. Earthworms (Eisenia fetida (Savigny)) were exposed to artificial soils spiked with Cd, Pb, Zn, and a Cd-Pb-Zn equitoxic mixture to estimate lethal CBRs and mixture toxicity. To evaluate the CBR developed for Zn, earthworms were also exposed to Zn-contaminated field soils receiving three different remediation treatments. Earthworm metal concentrations were measured using a procedure devised to isolate toxicologically active metal burdens via separation into cytosolic and pellet fractions. Lethal CBRs inducing 50% mortality (LD50, 95% CI) were calculated to be 5.72 (3.54-7.31), 3.33 (2.97-3.69), and 8.19 (4.78-11.6) mmol/kg for Cd, Pb, and Zn, respectively. Zn concentrations of dead earthworms exposed to a lethal remediated Zn-smelter soil were 3-fold above the LD50 for Zn and comparable to earthworm concentrations in lethal Zn-spiked artificial soils, despite a 14-fold difference in total soil Zn concentration between lethal field and artificial soils. An evaluation of the acute mixture toxicity of Cd, Pb, and Zn in artificial soils using the Toxic Unit (TU) approach revealed an LD50 (95% CI) of 0.99 (0.57-1.41) TU, indicating additive toxicity. Total Cd, Pb, and Zn concentrations in earthworms were good indicators of lethal metal exposure, and enabled the calculation at LD50s for lethality. The Zn-LD50 developed in artificial soil was applicable to earthworms exposed to remediated Zn-smelter soil, despite a 14-fold difference in total soil Zn concentrations. Mixture toxicity evaluated using LD50s from each single metal test indicated additive mixture toxicity among Cd, Pb, and Zn. Fractionation of earth worm tissues into cytosolic and pellet digests yielded mixed results for detecting differences in exposure at the sublethal level CBRs are useful in describing acute Cd, Pb, and Zn toxicity in earthworms, but linking sublethal exposure to total and/or fractionated residues may be more difficult. More research on detoxification, regulation, and tissue and subcellular partitioning of heavy metals in earthworms and other invertebrates is needed to establish the link between body residue and sublethal exposure and toxicity. Keywords: Bioavailability; Cd; critical body residues; earthworms; metals; Pb; soil; Zn

Evidence for the presence of glutamatergic receptors in adult Schistosoma mansoni

Several studies have suggested that l-glutamate is a putative neurotransmitter in helminths. The present study investigated the presence of non- N-methyl- d-aspartate (NMDA) ionotropic receptors for glutamate in four subcellular fractions from adult male Schistosoma mansoni. Low-affinity ( K d =221±80 nM) binding sites for [ 3 H ]kainic acid (KA) were detected in the heterogeneous (P 1) fraction, which contains pieces of unbroken worm tissues, tegument, nuclei, and some vesicles. This binding was inhibited by classical glutamatergic ligands in the following order of potency: KA> l- glutamate> α -amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA)>quisqualate≅6,7-dinitroquinoline-2,3-dione (DNQX). However, neither NMDA, a selective agonist for NMDA receptors, nor dl- threo-β-hydroxyaspartate (THA) and 1- trans-pyrollidine-2-dicarboxylic acid (PDC), inhibitors of high-affinity glutamate transporters, modified [ 3 H ]KA binding to the P 1 fraction. In addition, no specific binding for 10 nM [ 3 H ]AMPA was detected in any subcellular fraction from S. mansoni. These results suggested the presence of KA receptors in adult male worms. This is supported by the evidence that direct application of 10 μM KA to whole worms produced a corkscrew-like coiling of their bodies, modifying the motility of the worms. The KA-induced response, measured as a decrease of the body area, was time-dependent and reversible. PDC was ineffective at blocking the KA effects, indicating that KA does not depend on high-affinity glutamate transporters to reach its site of action. On the other hand, DNQX, the non-NMDA antagonist, was able to partially inhibit KA-induced responses. As a whole, the present data support the presence of a glutamatergic signaling pathway in this parasite.

Earthworms as bioindicators of mercury pollution from mining and other industrial activities

Mercury (Hg) can be released into the environment by various natural and industrial processes. Given the potential for environmental discharges from a number of sources and the severity of hazards associated with this highly toxic metal, potential mercury transformations must be well understood to effectively predict and prevent harmful human and environmental health effects. Bioindicators play an important role in identifying the factors controlling Hg toxicity and bioavailability and can ultimately be used to evaluate hazardous situations. A methodology using the earthworm Eisenia foetida has been developed to assess Hg bioavailability in mine tailings and aqueous solutions. Results indicate that E. foetida accumulate Hg and a positive correlation exists between Hg concentrations in worm tissues, the substrate they consume and length of exposure. To investigate the effect of natural organic acids on Hg bioavailability, metallic Hg (Hg 0 ) was dissolved in tannic acid and ‘fed’ to the worms in a substrate of paper and silica sand. Total Hg and methylmercury (MeHg) were analysed to determine whether methylation of Hg was occurring in the substrate, directly within worm intestines, or in the tannic acid–Hg solution. The MeHg:total Hg ratio was up to 160 times higher in worm tissues than both the tannic acid–Hg solution and the substrate. This result is particularly significant in organic-rich systems, where naturally occurring organic acids may be facilitating methylation within organisms digestive tracts.

Sulphur sources for chemoautotrophic nutrition of shallow water vestimentiferan tubeworms in Kagoshima Bay

To elucidate the sulphur sources for chemoautotrophy by the symbiotic bacteria of a vestimentiferan tubeworm, Lamellibrachia satsuma, living in Kagoshima Bay at depths of 80–100 m, we analysed the sulphur isotopic ratios of the animal tissues and compared them with environmental sulphur species collected in the field. Animals that had been maintained in an aquarium for over a year and supplied a known sulphur source were also investigated. The gas emitted from volcanic source in Kagoshima Bay contained rather heavy sulphide (+12·7 to +22·9‰ δ34S) compared with deep-sea hydrothermal vent systems (0 to +5‰). The tissue of the tubeworms contained very light sulphide (−21·5 to −25·9‰). It is inferred from the analysis of the aquarium-maintained specimens that the fractionation by the tubeworm or its symbiont was &lt;1·5‰. The sulphur source assimilated by the tubeworms in the field is therefore inferred to have δ ratio −19·1 to −24·6‰. This means that only 9·7 to 25·0% of the sulphur in the worm tissues can be derived from the volcanic gas and the rest must come from other sources, such as microbial activity in the bottom sediment.

Ultrastructure of cultured cells from Schistosoma japonicum

Ultrastructures and their dynamic changes of the cultured cells from Schistosoma japonicum were observed in the present experiments. Several types—including polygonal, round granular, deltaic fan-shaped and flagellated cells—were found in the cultures. The polygonal cells took a major ratio in the cultures from adult S. japonicum, while the majority from schistosomula was round granular cells. The ultrastuctures on the cell surface were different between the cells from adults and schistosomula. Some papilla-like tubercula, microvilli and pinocytotic vesicles were observed on the surface of adult cells, but none were found on schistosomula cells. However, more or less mitochondria, endoplasmic reticula, ribosomes and glycogen were observed in the cytoplasm of the cultured cells from both adults and schistosomula. Golgi complexes were rarely found. The nucleus was round, with round nucleolus inside and clear pores on the unit membrane. There was much lumpish heterochromatin located near to the nuclear membrane. Cells from different worm tissues had their own organelles. The germ cells, vitelline cells, flame cells, multinucleate subtegumental cells and nerve cells could be observed in the cultures from adults. The vitelline cells were the greatest in number and nerve cells were the least in number among them. Similarly, there were germ cells, sustentacular cells, flame cells, nerve cells, mast cells, muscle cells, multinucleate subtegumental cells, interstitial cells and penetration gland cells in the cultures from the schistomomula. In addition, a few division cells were also found. It indicated that the schistosomula cells had greater potential ability to proliferate than the adult cells in in vitro culture. Along with the prolongation of the culture time, degeneration of schistosomal cell occurred more and more. Generally, the electron density of cultures gradually got lower, the cristae of mitochondria blurred and disappeared and the mitochondria themselves swelled and finally vacuoled completely. Vitelline cells were most sensitive to the changes of the in vitro condition in all cultures. Their degeneration showed the following characteristics: (1) vitelline globules fused each other, the space between vitelline globules and the membrane surrounding them broadened gradually and vitelline globules were released and uncovered; (2) rough-surfaced endoplasmic reticula enlarged, vacuolated and the ribosomes dropped; and (3) the number and volume of lipid increased. The ultrastructural changes of most of the cultures from schistosomula had the following trends: (1) heterochromatin increased and euchromatin decreased gradually; and (2) endoplasmic reticula changed into short tubes and vacuoles and disappeared finally. The degenerative process of the cultures from S. japonicum consisted of necrosis according to the ultrastructural changes of the mitochondria, vitelline globules, chromatin and endoplasmic reticula within the cells. The changes of the above structures could be used to estimate whether the culture conditions were appropriate.

One in the eye for river blindness

This Journal Club article also appears in Trends in ParasitologyChemotherapy is the principal intervention in the control of filariasis. For Onchocerca volvulus, the species responsible for ‘river blindness’, the current approach is to treat with ivermectin (Mectizan), a drug highly effective at clearing the juvenile microfilarial stage of the parasite, which lives in the skin awaiting transmission by the blackfly vector. Unfortunately, Mectizan has no effect on the survival of adult worms, which can live for a decade or more in subcutaneous nodules, giving birth to millions of microfilariae. Consequently, treatment has to be repeated on an annual basis for several years and cover sufficient numbers of people to have any hope of interrupting transmission. For years, scientists have been searching for that elusive drug that can kill or permanently sterilize the long-lived adult nematodes, but with little success. Recently, a new target for anti-filarial chemotherapy has emerged from the revelation that intracellular Wolbachia bacteria have evolved an obligate symbiosis with their nematode hosts. Treatment of filarial nematodes with the antibiotics tetracycline or rifampicin leads to the depletion of Wolbachia, and results in long-term sterility and inhibition of worm development and viability. In the few years since this discovery, the first clinical trial to test the effect of antibiotics on the depletion of Wolbachia in human onchocerciasis has been completed in Ghana 1xDepletion of Wolbachia endobacteria in Onchocerca volvulus by doxycycline and microfilaridermia after ivermectin treatment. Hoerauf, A. et al. Lancet. 2001; 357: 1415–1416Abstract | Full Text | Full Text PDF | PubMed | Scopus (167)See all