Trace metals accumulation in the eco-system water - soil - vegetation (Agropyron cristatum) - common voles (Microtus arvalis) - parasites (Hymenolepis diminuta) in Radnevo region, Bulgaria.

BACKGROUND: Coal and coal processing industries provoke trace metal pollution, which has a negative effect on the water - soil - vegetation - small mammals eco-system, constituting part of the food chain and exerting a serious impact on human health. OBJECTIVES: Assessment of the environmental impact of Maritza iztok coal complex, situated east of Radnevo town, Bulgaria, by tracking the dynamics and accumulation of trace metals in the eco-system water - soil - vegetation - common voles - parasites. METHODS: Samples from surface waters, their nearby uncultivated soils, meadow uncultivated vegetation (Agropyron cristatum) and field common voles (Microtus arvalis) were collected. In situ measurements and laboratory extraction procedures and analyses were performed. Accumulation and mobility indices were calculated. The distribution of dissolved Mn, Ni, Cu, Zn and Pb chemical species in water-soil extracts was calculated using a thermodynamic approach. Two thermodynamic models were applied - the classical ion-association model for calculating the inorganic trace metal species and the Stockholm Humic Model (SHM) accounting for the complexation reactions of trace metals with organic matter. Visual Minteq computer program, Version 3.1 was used. The relationship chemical species - bioaccumulation was discussed. RESULTS: Pb and Mn, together with SO42- and PO43- were found to be the main pollutants of waters in the region. The soils studied exhibited low concentrations of trace metals, not exceeding the specified MACs. The content of Mn was the highest, followed by Zn, Pb, Cu and Ni. The highest phytoaccumulation coefficients in the studied uncultivated grass vegetation were calculated for Cu and Zn, being 1-2 orders of magnitude higher than those of Mn and Ni. The accumulation of trace metals was explained on the basis of ions mobility and chemical species distribution. In the case of the host-parasite system Microtus alvaris - Hymenolepis diminuta, Zn displays the highest accumulation coefficient, followed by those of Cu and Pb. The parasite showed a higher bioaccumulation compared to infected common voles, with the highest bioaccumulation found for Ni. CONCLUSIONS: The bioaccumulation of trace metals depends on their mobility, concentration and chemical forms in water-soil solutions. Metal-organic species stimulate the phytoaccumulation of trace metals while inorganic ones suppress it. The sequence of trace metals bioaccumulation in common voles is analogous to that of soil contamination. The parasite exhibited higher bioaccumulation levels compared to infected common voles.

Dynamics of trace metals in the system water - soil - plant - wild rats - tapeworms (Hymenolepis diminuta) in Maglizh area, Bulgaria.

BACKGROUND: The impact of chemical elements on the biosphere is a function of their concentration and chemical form. Elucidation and prognosing of the latters in water basins and soil extracts is of particular significance for the assessment of their bioaccumulation in plants and animals. OBJECTIVES: Trace metals dynamics in the system water - soil-plant-wild rats - Hymenolepis diminuta in two agro-industrial zones (East and West) around Maglizh city, Bulgaria were investigated through experimental studies and thermodynamic modelling of the chemical species. METHODS: Samples from surface waters of rivers, their nearby uncultivated soils, meadow uncultivated vegetation (Ranunculus acris and Gramineae) and field rats were collected. In situ measurements and laboratory analyses were performed for the determination of the physico-chemical characteristics and total concentrations of Al, Fe, Mn, Ni, Cu, Zn and Pb. The distribution of their dissolved chemical species in water samples and in the aqueous soil extracts was calculated using a thermodynamic approach. The relationship chemical species - bioaccumulation was discussed. RESULTS: Waters and soils in the East zone of Maglizh area were found to be more polluted compared to those in the West one, regarding Ni, Mn, Zn, Pb and Cu, while Mn and Cu displayed the highest mobility in West zone soils. Trace metals contents in Ranunculus acris exceed that in Gramineae, since the highest accumulation factors were calculated for Cu and Zn. The highest accumulation in rats was found for Zn followed by Cu, being higher in the West zone. Thermodynamic modelling shows that Mn2+ free ions are dominant in both waters and aqueous soil extracts. Ni2+ and Zn2+ ions followed by metal-organic complexes are dominant in waters of East zone while metal-organic complexes followed by free ions are dominant in waters of West zone and both soil extracts. Metal-organic complexes are dominant for Fe, Cu and Pb in all samples studied, while mainly hydroxy forms (Al(OH)4-) followed by metal-organic complexes are typically for Al depending on pH. CONCLUSIONS: Experimentally established bioaccumulation of trace metals in the studied vegetation and rats is a consequence of the total concentration of trace metals in waters and soils, their mobility and chemical species. The dominance of organic complexes of trace metals is a prerequisite for their bioaccumulation in plants. Rats are in direct contact with the soil solution and therefore, of importance is the content of free ions of Mn2+, Ni2+, Zn2+, which are easily absorbed through the skin. The host-helminth system wild rat/H. diminuta could be used as a bioindicator for trace metals pollution.

Analysis of lead pollution levels within an urban ecosystem using the cestode Hymenolepis diminuta and its rat hosts as bioindicators.

The overall goal of this study was to use the Rattus spp./Hymenolepis diminuta model to assess environmental lead pollution in different landscape units of an urban ecosystem. Rats of the genus Rattus were collected from three shanty towns and three residential neighbourhoods of the city of Buenos Aires. Concentrations of lead in the livers of wild rats and in their parasite H. diminuta were measured using inductively coupled plasma mass spectrometry (ICP-MS). The landscape unit and tissue type had a significant effect on lead concentration, being higher in residential neighbourhoods as well as in H. diminuta tissue. Nevertheless, no significant differences were found for the mean lead concentration in livers between uninfected and infected rats. Since the available information describing heavy-metal pollution within the city of Buenos Aires is scarce, the results of this study allow us to update data about the extent of biologically available lead contamination. Considering that rats and H. diminuta are distributed worldwide, this monitoring system for lead pollution might be applied successfully in other urban ecosystems.

Identification of immunogenic proteins of the cysticercoid of Hymenolepis diminuta.

BACKGROUND: A wide range of molecules are used by tapeworm metacestodes to establish successful infection in the hostile environment of the host. Reports indicating the proteins in the cestode-host interactions are limited predominantly to taeniids, with no previous data available for non-taeniid species. A non-taeniid, Hymenolepis diminuta, represents one of the most important model species in cestode biology and exhibits an exceptional developmental plasticity in its life-cycle, which involves two phylogenetically distant hosts, arthropod and vertebrate. RESULTS: We identified H. diminuta cysticercoid proteins that were recognized by sera of H. diminuta-infected rats using two-dimensional gel electrophoresis (2DE), 2D-immunoblotting, and LC-MS/MS mass spectrometry. Proteomic analysis of 42 antigenic spots revealed 70 proteins. The largest number belonged to structural proteins and to the heat-shock protein (HSP) family. These results show a number of the antigenic proteins of the cysticercoid stage, which were present already in the insect host prior to contact with the mammal host. These are the first parasite antigens that the mammal host encounters after the infection, therefore they may represent some of the molecules important in host-parasite interactions at the early stage of infection. CONCLUSIONS: These results could help in understanding how H. diminuta and other cestodes adapt to their diverse and complex parasitic life-cycles and show universal molecules used among diverse groups of cestodes to escape the host response to infection.

The identification of risk and essential elements along the strobila of the rat tapeworm Hymenolepis diminuta.

The rat tapeworm Hymenolepis diminuta can bioconcentrate several elements to conspicuously higher concentrations than tissues of their definitive host. The main aim of this study was to locate parts of the tapeworm into which lead, cadmium, zinc, copper, manganese and iron are accumulated. Male Wistar rats were experimentally infected with H. diminuta and worms were exposed to two different forms of lead for 6 weeks through the oral exposure of their rat hosts. After the exposure period, the element levels were determined in the posterior and anterior proglottids of the tapeworm. In all cases, lead concentrations were higher in the anterior parts than the posterior parts. Concentrations of cadmium, copper, iron, manganese and zinc were also significantly higher in the anterior parts. Zinc concentrations showed an opposite trend, with higher zinc levels detected in the posterior part of the strobila, in the control group. The present study demonstrates that risk and essential elements are accumulated mainly into the anterior part of H. diminuta.

A method for measuring the attachment strength of the cestode Hymenolepis diminuta to the rat intestine.

A unique adaptation of many internal parasites of mammals is their ability to stay in the intestine for extended periods of time and resist the normal peristaltic movements and forces that push and expel material. To better understand parasite adhesion behaviour and replicate their attachment method in medical devices, an experiment was designed and performed using the rat tapeworm, Hymenolepis diminuta. The experiment employed a tensile test machine and a digital scale and was designed to calculate the attachment strength of the scolex to the mucosa through the change of the value of the digital scale during the tensile test. The attachment force of H. diminuta is 0.021 ± 0.011 g. This method could be applied in studies of parasite biomechanics and the results may help medical device researchers to better mimic the unique functional morphology of this species of parasite.

Mass spectrometry analysis of the excretory-secretory (E-S) products of the model cestode Hymenolepis diminut a reveals their immunogenic properties and the presence of new E-S proteins in cestodes.

Hymenolepis diminuta is an important model species in studies of therapeutics, biochemical processes, immune responses and other aspects of cestodiasis. The parasite produces numerous excretory-secretory (E-S) proteins and a glycocalyx covering its body. Our study focused on the mass spectrometry analysis of the E-S material with an objective to determine if E-S contains any new proteins, in particular those that can be identified as: antigens, vaccine candidates and drug targets. These proteins might engage directly in host-parasite interactions. Adult parasites collected from experimentally infected rats were cultured in vitro for 5 and 18h. Immunoblotting was used to verify which E-S protein bands separated in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) react with specific antibodies from sera of infected rats. We identified thirty-nine proteins by LC-MS/MS (liquid chromatography mass spectrometry). Results indicated the presence of proteins that have never been identified in cestode E-S material. Immunoblotting showed the immunogenicity of E-S products of H. diminuta, most probably associated with the presence of proteins known as antigens in other flatworm species. Among identified proteins are those engaged in immunomodulatory processes (eg. HSP), in response to oxidative stress (peroxidasin) or metabolism (eg. GAPDH). The predominant functions are associated with metabolism and catalytic activity. This is the first study identifying E-S-proteins in adult tapeworms, thus providing information for better understanding host-parasite interrelationships, and may point out potential targets for vaccines or drug discovery studies, as among the proteins observed in our study are those known to be antigens.

Extracts of the rat tapeworm, Hymenolepis diminuta, suppress macrophage activation in vitro and alleviate chemically induced colitis in mice.

Analysis of parasite-host interactions can reveal the intricacies of immunity and identify ways to modulate immunopathological reactions. We assessed the ability of a phosphate-buffered saline-soluble extract of adult Hymenolepis diminuta to suppress macrophage (human THP-1 cell line, murine peritoneal macrophages) activity in vitro and the impact of treating mice with this extract on colitis induced by dinitrobenzene sulfonic acid (DNBS). A high-molecular-mass fraction of adult H. diminuta (HdHMW) or excretory/secretory products reduced macrophage activation: lipopolysaccharide (LPS)-induced interleukin-1beta (IL-1beta), IL-6, and tumor necrosis factor alpha (TNF-alpha) and poly(I:C)-induced TNF-alpha and IL-6 were suppressed by HdHMW. The active component in the HdHMW extract was minimally sensitive to boiling and trypsin digestion, whereas the use of sodium metaperiodate, as a general deglycosylation strategy, indicated that the immunosuppressive effect of HdHMW was at least partially dependent on a glycan: treating the HdHMW with neuraminidase and alpha-mannosidase failed to inhibit its blockade of LPS-induced TNF-alpha production by THP-1 macrophages. Mice treated with DNBS developed colitis, as typified by wasting, shortening of the colon, macroscopic and microscopic tissue damage, and an inflammatory infiltrate. Mice cotreated with HdHMW (three intraperitoneal injections) displayed significantly less inflammatory disease, and this was accompanied by reduced TNF-alpha production and increased IL-10 and IL-4 production by mitogen-stimulated spleen cells. However, cotreatment of mice with neutralizing anti-IL-10 antibodies had only a minor impact on the anticolitic effect of the HdHMW. We speculate that purification of the immunosuppressive factor(s) from H. diminuta has the potential to lead to the development of novel immunomodulatory drugs to treat inflammatory disease.