The oral bioavailability of soil-borne risk elements for small terrestrial mammals: Microtus arvalis (Pallas, 1778) and Apodemus sylvaticus L. and its implication in environmental studies.

The aims of this study were (i) to specify real risk elemetnt (RE) uptake by wild terresrial mammals (A. sylvaticus and M. arvalis), (ii) to describe RE distribution in critical organs such as the liver and kidney, and (iii) to determine potencial differences in RE toxicokinetics with regards to individual species or sex. Three groups of experimental animals were fed diets amended with soil and green biomass of hyperaccumulator Arabidopsis halleri with different RE levels. The contents of As, Cd, and Pb in the liver and kidneys of the animals reflected the element contents in the diet. Higher Cd and Pb accumulation ability was observed in A. sylvaticus compared to M. arvalis tissues, and an opposite pattern was recorded for As. Zn contents in tissues of both species remained unchanged, and total contents of Zn in the exposed animals even tended to decrease compared to the controls. Results of this study indicate a generally similar response of both species to elevated RE contents in the diet, confirming these wild rodents as suitable models for RE biomonitoring in ecosystems. However, our data highlights some distinction in As toxicokinetics in wood mice compared to that of field voles and a significantly higher accumulation of Pb and Cd in females. Therefore, factors of species and sex should not be overlooked if relevant data are to be obtained in environmental studies conducted on small terrestrial rodents.

Molecular cytogenetic analysis of a triploid population of the human broad tapeworm, Dibothriocephalus latus (Diphyllobothriidea).

The large-sized tapeworm Dibothriocephalus latus is known as the broad or fish-borne cestode of mammals that is capable to infect humans and cause diphyllobothriosis. Recently, molecular data on D. latus has been accumulating in the literature and a complete genome sequence has been published; however, little is known about the karyotype and chromosome architecture. In this study, an in-depth karyological analysis of 2 D. latus specimens was carried out. The plerocercoids originated from a perch caught in subalpine Lake Iseo (Italy) and the tapeworms were reared in hamsters. Both specimens contained cells with a highly variable number of chromosomes ranging from18 to 27. Nevertheless, the largest portion of mitotic figures (47%) showed a number corresponding to the triploid set, 3n = 27. Accordingly, the karyotype of the analyzed specimens consisted of 9 triplets of metacentric chromosomes. Fluorescence in situ hybridization (FISH) with the 18S rDNA probe clearly demonstrated the presence of 3 clusters of hybridization signals on the triplet of chromosome 7, thus confirming the triploid status of the specimens. FISH with a telomeric (TTAGGG)n probe confined hybridization signals exclusively to the terminal chromosomal regions, supporting the earlier findings that this repetitive motif is a conserved feature of tapeworm telomeres.

The response of soil nematode Caenorhabditis elegans on the sewage sludge-derived micropollutants.

Sewage sludge application to soil is of great interest, due to required organic matter and the wide spectra of nutrients it provides. However, the presence of unpredictable content of emerging contaminants may turn this valuable raw material into a hazardous substance. In this study, three selected sewage sludges derived micropollutants from different origins; that is, one each under persistent organic pollutants (POPs), pharmaceuticals and personal care products (PPCPs) were considered. The effect of each micropollutant on the feeding activity of free-living soil nematode Caenorhabditis elegans was analysed. The analysis was performed in model soil solution using a larval feeding inhibition assay. The results showed no significant effects from selected POP-2,2',4,4',5-pentabromodiphenyl either and pharmaceutical-chlortetracycline on the feeding activity of tested nematodes. On the contrary, feeding activity was inhibited by PPCP-galaxolide (HHCB) with an effective concentration of 12.2 ±â€¯2.2 mg.l-1. The calculated risk quotient for galaxolide (RQ = 0.14) demonstrated a medium ecological risk to the nematodes. Based on our findings, concentrations of micropollutants in sewage sludge treated soil pose negligible risk to feeding activity of soil nematode. However, the potential impact of musk compounds on free-living soil biota requires detailed evaluation in further research.

Mobility and bioaccessibility of risk elements in the area affected by the long-term opencast coal mining.

The potential environmental hazards of risk elements in the area affected by the opencast coal mine and/or coal combustion for plants and animals was assessed by using a suite of laboratory bioaccessibility tests. The chosen sampling area was in the vicinity of the largest coal mine spoil in the Sokolov coal basin (Czech Republic). For an estimation of the oral bioaccessibility of the risk elements in soils, the physiologically based extraction tests were applied. Among the available methods for estimating the pulmonary bioaccessibility of elements, the Gamble's and Hatch's tests were chosen. The results showed elevated pseudo-total soil contents of As, Be, Cd, Cu, Pb, V, and Zn. Among these elements, only Cd showed substantial bioaccessibility for plants, as documented by the high Risk Assessment Code, reaching up to 47%, and the highest plant-availability, where the maximum Bioaccumulation Factor in plants reached up to 4.5. The simulated body fluids showed the highest bioaccessibility of Cd, but also substantial bioaccessible pools of As and Be, the elements frequently found at the brown coal mining and processing areas. For better understanding of the risk element bioaccessibility under the specific conditions, the released element pools should be related to the particular soil physicochemical parameters.

Variability of trace element distribution in Noccaea spp., Arabidopsis spp., and Thlaspi arvense leaves: the role of plant species and element accumulation ability.

Laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) was applied for the determination of Cd and Zn distributions within the leaves of Cd- and Zn-hyperaccumulating plants, Noccaea caerulescens, N. praecox, and Arabidopsis halleri, in contrast to nonaccumulator species, Thlaspi arvense and A. thaliana. The elemental mapping of the selected leaf area was accomplished via line scans with a 110-µm-diameter laser beam at a 37-µm s-1 scan speed and repetition rate of 10 Hz. The lines were spaced 180 µm apart and ablated at an energy density of 2 J cm-2. The elemental imaging clearly confirmed that Cd was predominantly distributed within the parenchyma of the T. arvense, whereas in the Noccaea spp. and A. halleri, the highest intensity Cd signal was observed in the veins of the leaves. For Zn, higher intensities were observed in the veins for all the plant species except for A. thaliana. Close relationships between Zn and Ca were identified for the Noccaea spp. leaves. These relationships were not confirmed for A. halleri. Significant correlations were also proved between the Cd and Zn distribution in A. halleri, but not for the Noccaea spp. For both T. arvense and A. thaliana, no relevant significant relationship for the interpretation of the results was observed. Thus, the LA-ICP-MS imaging is proved as a relevant technique for the description and understanding of the elements in hyperaccumulating or highly accumulating plant species, although its sensitivity for the natural element contents in nonaccumulator plant species is still insufficient.

Metabolic transformation and urinary excretion of selenium (Se) in rats fed a Se-enriched defatted rapeseed (Brassica napus, L.) diet.

Se-Enriched defatted rapeseed (Se-DRS), a by-product of rapeseed oil production, is high in Se. This study aimed to detect possible changes in Se-metabolism after Se-DRS consumption and to assess its suitability as a Se supplement in monogaster feed. In this experiment, rats were fed three types of diets differing in Se-form: soybean meal, Se-salt and Se-DRS. Potential differences in Se-metabolism were evaluated based on the total Se concentrations along with non-protein Se speciation patterns in the urine and blood serum of the rats. The total Se concentrations were determined using inductively coupled plasma-mass spectrometry (ICP-MS). Se compounds in the samples were identified through high-performance liquid chromatography/ICP-MS. The total Se excretion rate was dose-dependent, and excess of dietary Se was eliminated in the form of Se-sugar 1, regardless of the administered Se-form. The excretion dynamics of dietary selenomethionine (soybean meal) and methylselenocysteine (Se-DRS) were different to that of the mineral Se (Se-salt). The proportions of Se species in the urine were significantly influenced by the Se source. A specific metabolic pathway, resulting in urinary excretion of Se-sugar 2, was observed in animals fed Se-DRS. Neither the total Se concentrations nor the serum Se speciation patterns differed between the animals fed the conventional diet and those fed the Se-DRS-enriched diet. Based on these results, Se-DRS can be considered a suitable Se supplement for monogaster nutrition in Se-deficient regions.

Efficacy and persistent activity of moxidectin against natural Muellerius capillaris infection in goats and pathological consequences of muelleriosis.

The effect of moxidectin against natural Muellerius capillaris infection in goats was evaluated in this study. Long-acting moxidectin at a single dose of 1 mg kg(-1) body weight was administered to an entire flock (n=10) of goats. The individual faecal larval count reduction was applied as an indicator of treatment efficacy. A significant reduction (>98%) in larval counts was observed in all surveyed animals 14 days after drug administration. Moxidectin demonstrated persistent activity in this study; the mean faecal larval count reduction was 99.1% ± 1.8 on day 77 of the treatment. Macroscopic abnormalities and histological changes in the lungs of two infected goats were evident during the post-mortem examination. The pathological consequences of M. capillaris infection were observed even three months after parasite elimination. The results of this study indicate that moxidectin is a highly effective anthelmintic agent for the control of muelleriosis in goats. This drug provides animals with fifteen weeks of protections against M. capillaris reinfection.

Bioaccessibility versus bioavailability of essential (Cu, Fe, Mn, and Zn) and toxic (Pb) elements from phyto hyperaccumulator Pistia stratiotes: potential risk of dietary intake.

Aquatic weeds are widely used as animal feed in developing countries. However, information about element bioavailability from these plants is lacking. A combination of an in vitro method [physiologically based extraction test (PBET)] and an in vivo feeding trial was used in this study to investigate potential element bioaccessibility and estimated bioavailability of Pistia stratiotes (PS). Cu, Fe, Mn, Zn, and Pb concentrations in PS biomass, artificial gastrointestinal fluids, and rat tissues were determined using atomic absorption spectrometry with electrothermal atomization and inductively coupled plasma-atomic emission spectrometry. PS exhibited elevated Fe, Mn, and Pb levels. The PBET revealed high bioaccessibility of all monitored elements from PS biomass. The results of the in vivo trial were inconsistent with those of the PBET, because animals fed PS exhibited low levels of essential elements in the tissues. The consumption of a PS-supplemented diet significantly decreased total Fe levels and increased the total level of accumulation of Pb in exposed animals. Significantly reduced amounts of essential elements in the intestinal walls indicated a potential disruption in nutrient gastrointestinal absorption in animals fed PS.

Arrested development of experimental Cyathostominae infections in ponies in Czech republic.

Nine ponies, aged 12 months at the time of infection, were randomly allocated to one of three groups. Each pony was infected with 30,000 infective cyathostome larvae stored 4 weeks under the natural environmental conditions of the Czech Republic. Horses of Group A were infected with larvae conditioned from August 16 to September 12 (the first phase of the experiment). Horses of Group B were inoculated with larvae conditioned from October 10 to November 4 (the second phase of the experiment). In the third phase, horses from Group C received larvae stored under natural conditions (from 8.11 to 18.11.2011). The larvae were then kept under laboratory conditions (5 °C, the light reduction according to the nature condition). In the first and second phase of the experiment, 98.57% and 95.48% of adult nematodes were found, respectively. However, only 29.07% of adults were found in the third phase of experiment, and the proportion of arrested larvae (EL3) was 67.73%. The share of these larvae in the cecum during last phase of the experiment comprised 92.59% of all cyathostomes.

Is the tapeworm able to affect tissue Pb-concentrations in white rat?

The effect of gastrointestinal helminths on Pb accumulation in the host body is ambiguous. A laboratory experiment with Rattus norvegicus/Hymenolepis diminuta model was conducted to determine Pb toxicokinetics in a terrestrial host-parasite system. The ET-AAS or ICP-OES techniques were used to determine Pb concentrations (CPb) in both tapeworms and host tissues (kidney, liver, bone, testes, muscle and intestinal wall). Concerning the entire host-parasite system, the highest CPb were detected in H. diminuta. Rat kidneys and bone were the only two tissues whose mean Pb levels were lower in parasitized animals than they were in non-infected subjects after both levels of exposure. At low Pb exposure, parasitization slightly changed the Pb toxicokinetics in the host body. However, with respect to tissue at the same exposure level, no significant differences were detected between the parasitized and non-parasitized animals and no significant correlations were found between CPb in tapeworms and those of host tissues. The results of this study indicate that H. diminuta does not protect rat from elevated Pb exposure even if tapeworm accumulates a higher portion of ingested Pb dose compared with that of the most Pb-loaded host soft tissue. The portion of Pb dose accumulated in H. diminuta correlates positively with parasite biomass.

Faecal excretion dynamic during subacute oral exposure to different Pb species in Rattus norvegicus.

Faecal excretion is a basic means of detoxification upon ingestion of Pb-contaminated feed. In order to determine a time course of Pb elimination after oral exposure to two different forms of this heavy metal (lead acetate vs. phyto-bound Pb), a feeding study was carried out in experimental rats using the Pb phyto-hyperaccumulator Pistia stratiotes as a model diet. The effect of starvation on Pb excretion was further studied in rats that were fed plant material. Twelve Pb doses (7 µg Pb/1 g BW) were administered orally over a 5-week period. Faeces samples were collected 24 and 72 h post-exposure. Inductively coupled plasma optical emission spectrometry and electrothermal absorption spectrometry methods were used for determination of heavy metal concentrations. Up to 53 % of ingested Pb was rapidly eliminated from the exposed rats via faeces within 24 h after exposure. Faecal excretion in exposed rats differed significantly when compared to that of the control group. Fasting before exposure reduced Pb excretion by up to 50 %. Faecal excretions of both examined Pb forms exhibited almost identical patterns. Considerable differences were revealed concerning total excretion levels; lead acetate was excreted in amount greater extent than those of phytobound Pb. Results of our study suggest that Pb forms occurring in the P. stratiotes tissues are absorbed through the gastrointestinal tract to a greater extent than Pb from lead acetate. Therefore, higher portions of ingested Pb can be available for potential accumulation in tissues of exposed subjects.

Concentrations of Zn, Mn, Cu and Cd in different tissues of perch (Perca fluviatilis) and in perch intestinal parasite (Acanthocephalus lucii) from the stream near Prague (Czech Republic).

We monitored concentrations of Cd, Cu, Mn and Zn in acantocephalan parasites (Acanthocephalus lucii) and its final host (Perca fluviatilis). The concentrations in parasites were found to be significantly higher than those found in the muscle, gonads and liver of fish host. The bioaccumulation factor values (BF=C(parasite)/C(host muscle)) were 194, 24.4, 2.2 and 4.7 for Cd, Cu, Mn and Zn, respectively. This suggests a benefit for the host due to the high accumulation of toxic cadmium.

Which McMaster egg counting technique is the most reliable?

This study focuses on the comparison of three selected modifications of the McMaster counting technique, namely the McMaster method modified by Wetzel (W) and Zajícek (Z), as well as the concentration McMaster technique according to Roepstorff and Nansen (R&N). These modifications differ in the weights of faeces examined (W, 2 g/Z, 1 g/R&N, 4 g), flotation solutions (W, NaCl/Z, MgSO(4) + Na(2)S(2)O(3)/R&N, NaCl + glucose), centrifugation (W, none/Z, 2,000 RPM for 2 min and 2,000 RPM for 1 min/R&N, 1,200 RPM for 5 min), number of McMaster chambers investigated (W, 3/Z, 2/R&N, 2), and multiplication factors used (W, 67/Z, 33/R&N, 20). To investigate the sensitivity and reliability of these methods, nematode eggs (Teladorsagia circumcincta) were used. Parasite elements are distributed through negative binomial distribution in naturally infected host faeces, and the number of parasite elements in a given amount of faeces sample is unknown to man. Therefore, we decided to prepare the exact number of eggs which were added to the parasite negative faeces; the faecal sample was then investigated. From this perspective, this is the first time a comparison of the McMaster methods has been so accurately investigated. This approach allows us to evaluate the real sensitivity and reliability of the tested method. As the findings of this study indicate, the highest sensitivity and reliability were obtained using the Roepstorff and Nansen modification. This McMaster modification is able to detect 20 eggs per sample (in 70% of samples). Concentrations of 200 and 500 eggs can be found in almost 100% of samples. Moreover, this method is simple, cheap and fast. For these reasons, we can recommend this method for routine veterinary practice.

Intestinal parasite Acanthocephalus lucii (Acanthocephala) from European perch (Perca fluviatilis) as a bioindicator for lead pollution in the stream "Jevanský potok" near Prague, Czech Republic.

Lead concentrations in the tissues of perch and its parasites were determined as mg/kg dw. Lead was found at higher concentrations in the acanthocephalans (11.56) than in different tissues (liver, gonads and muscle with skin and bone) of perch. With respect to fish tissues, the highest concentrations of lead were present in the liver (1.24), followed by the gonads (0.57) whereas the lowest concentrations were in the muscle with skin and bone (0.21). The bioconcentration factors for lead indicated that parasites accumulate metals to a higher degree than fish tissues--lead concentrations in acanthocephalans were 9.32, 19.27 and 55.05 higher than in liver, gonads and muscles of host, respectively.