Sustainable use of composted sewage sludge: Metal(loid) leaching behaviour and material suitability for application on degraded soils.

Composted sewage sludge was investigated as a promising material for the reclamation or remediation of degraded sites. Using sewage sludge as soil amendment provides environmental benefits and risks while supporting circularity and waste minimisation. This study aims to comprehensively assess the suitability of locally available low-cost sludge treatment for sustainable and environmentally safe topsoil disposal in a brownfield area affected by coal mining. A nine-month composting was conducted before field application to the soil environment. The objectives were to assess: (i) composting time-dependent and pH-dependent metal(loid) leachability from composted sludges, (ii) the effect of sludges on metal(loid) leachability from soil over the first six months, and (iii) metal(loid) plant uptake during the first vegetation season as well as the bioaccumulation and translocation factors. The set of standardised leaching experiments confirmed the positive effect of compost maturity, i.e. despite some fluctuations over time, metal(loid) availability from the final composts was very low. Some metals showed unusual pH-dependent behaviour with the highest leachability at pH 8 due to excessive release of dissolved organic matter from the not-yet-stabilised matrix. Ecotoxicity testing confirmed the safety of the final composts for further soil application. The sludge-amended plots displayed similar metal(loid) leaching and pH evolution in time compared to the control biomass-amended plot. However, plant species (Artemisia vulgaris L.) that formed the natural vegetation cover of the experimental plots showed cumulative metal(loid) uptake. Cadmium and zinc were identified as the critical metals possibly related to the applied sludges, yielding high bioaccumulation and translocation factors. Yet, the quality of the compost feedstock, heterogeneity, and background values of the brownfield site need to be considered. Nevertheless, soil respiration indicated no adverse effects on soil health six months after sludge application. Overall, the composted material demonstrated potential suitability for remediation application in the studied area.

RADIOCARBON DATING OF CHARCOALS FROM HISTORICAL MORTARS FROM TÝROV AND PYSOLEC CASTLES.

Organic inclusions in lime binders provide useful samples for radiocarbon dating of historical objects. Two Czech castles Týrov and Pysolec from Late Middle Ages were explored, and tens of charcoals were found in their walls. The radiocarbon content of the charcoals was measured with accelerator mass spectrometry. The dating results showed that none of the charcoals were younger than the known historical ages (Týrov: 1260 - 1270, Pysolec: 1300 - 1340), but some were considerably older. Two charcoals from Pysolec castle dated to Palaeolithic, likely originating from fluvial sediments added as an aggregate to the mortar. When excluding these two charcoals, the others indicated most likely dates being 50-100 y older than the building dates of the castles. This systemic effect corresponds to the age of wood used for lime burning and shall be accounted for when dating mortars using charcoals.

Application of co-composted biochar significantly improved plant-growth relevant physical/chemical properties of a metal contaminated soil.

A woody-biochar was added to waste biomass during a composting process. The resulting compost-char was amended to a metal contaminated soil and two plant species, L. perenne and E. sativa, were grown in a pot experiment to determine 1) plant survival and stress factors, 2) uptake of metals to plants and, 3) chemical characteristics of sampled soils and pore waters. Compost supplemented with biochar after the composting process were also tested, as well as a commercially available compost, for comparison. Co-composting with biochar hastened the composting process, resulting in a composite material of reduced odour, increased maturity, circum-neutral pH and increased moisture retention than compost (increase by 3% of easily removable water content). When amended to the soil, CaCl2 extractable and pore water metals s were reduced by all compost treatments with little influence of biochar addition at any tested dose. Plant growth success was promoted furthest by the addition of co-composted biochar to the test soil, especially in the case of E. sativa. For both tested plant species significant reductions in plant metal concentrations (e.g. 8-times for Zn) were achieved, against the control soil, by compost, regardless of biochar addition. The results of this study demonstrate that the addition of biochar into the composting process can hasten the stability of the resulting compost-char, with more favourable characteristics as a soil amendment/improver than compost alone. This appears achievable whilst also maintaining the provision of available nutrients to soils and the reduction of metal mobility, and improved conditions for plant establishment.

PREPARATION OF WOOD, CHARCOAL AND BONE COLLAGEN MICRO SAMPLES USING AUTOMAT FOR AMS RADIOCARBON DATING.

For the pretreatment of wood, charcoal and collagen from bone micro samples using the Acid-Base-Acid (ABA) method, we have assembled an automated computer-controlled unit in our laboratory CRL. The sample is placed in a glass single-necked cuvette. The machine consists of prepared solutions which are guided through capillaries, switching valve and peristaltic pump into the cuvette with the sample according to the currently selected program. The automat can be used for the pretreatment of charcoal, wood and also collagen from bones.

Functional non-synonymous variants of ABCG2 and gout risk.

Objectives: Common dysfunctional variants of ATP binding cassette subfamily G member 2 (Junior blood group) (ABCG2), a high-capacity urate transporter gene, that result in decreased urate excretion are major causes of hyperuricemia and gout. In the present study, our objective was to determine the frequency and effect on gout of common and rare non-synonymous and other functional allelic variants in the ABCG2 gene. Methods: The main cohort recruited from the Czech Republic consisted of 145 gout patients; 115 normouricaemic controls were used for comparison. We amplified, directly sequenced and analysed 15 ABCG2 exons. The associations between genetic variants and clinical phenotype were analysed using the t-test, Fisher's exact test and a logistic and linear regression approach. Data from a New Zealand Polynesian sample set and the UK Biobank were included for the p.V12M analysis. Results: In the ABCG2 gene, 18 intronic (one dysfunctional splicing) and 11 exonic variants were detected: 9 were non-synonymous (2 common, 7 rare including 1 novel), namely p.V12M, p.Q141K, p.R147W, p.T153M, p.F373C, p.T434M, p.S476P, p.D620N and p.K360del. The p.Q141K (rs2231142) variant had a significantly higher minor allele frequency (0.23) in the gout patients compared with the European-origin population (0.09) and was significantly more common among gout patients than among normouricaemic controls (odds ratio = 3.26, P < 0.0001). Patients with non-synonymous allelic variants had an earlier onset of gout (42 vs 48 years, P = 0.0143) and a greater likelihood of a familial history of gout (41% vs 27%, odds ratio = 1.96, P = 0.053). In a meta-analysis p.V12M exerted a protective effect from gout (P < 0.0001). Conclusion: Genetic variants of ABCG2, common and rare, increased the risk of gout. Non-synonymous allelic variants of ABCG2 had a significant effect on earlier onset of gout and the presence of a familial gout history. ABCG2 should thus be considered a common and significant risk factor for gout.

Prevalence of URAT1 allelic variants in the Roma population.

The Roma represents a transnational ethnic group, with a current European population of 8-10 million. The evolutionary process that had the greatest impact on the gene pool of the Roma population is called the founder effect. Renal hypouricemia (RHUC) is a rare heterogenous inherited disorder characterized by impaired renal urate reabsorption. The affected individuals are predisposed to recurrent episodes of exercise-induced nonmyoglobinuric acute kidney injury and nephrolithiasis. To date, more than 150 patients with a loss-of-function mutation for the SLC22A12 (URAT1) gene have been found, most of whom are Asians. However, RHUC 1 patients have been described in a variety of ethnic groups (e.g., Arab Israelis, Iraqi Jews, Caucasians, and Roma) and in geographically noncontiguous countries. This study confirms our previous findings regarding the high frequency of SLC22A12 variants observed. Frequencies of the c.1245_1253del and c.1400C>T variants were found to be 1.92% and 5.56%, respectively, in a subgroup of the Roma population from five regions in three countries: Slovakia, Czech Republic, and Spain. Our findings suggested that the common dysfunction allelic variants of URAT1 exist in the general Roma population and thus renal hypouricemia should be kept in differential diagnostic algorithm on Roma patients with defect in renal tubular urate transport. This leads to confirm that the genetic drift in the Roma have increased the prevalence of hereditary disorders caused by very rare variants in major population.

Metallothionein-like peptides involved in sequestration of Zn in the Zn-accumulating ectomycorrhizal fungus Russula atropurpurea.

Homeostatic mechanisms preventing the toxicity of free Zn ions in cells involve, among others, cytosolic Zn-binding ligands, particularly the cysteine-rich metallothioneins (MTs). Here we examined the Zn-binding peptides of Russula atropurpurea, an ectomycorrhizal fungus known for its ability to accumulate high amounts of Zn in its sporocarps. The Zn complexes and their peptide ligands were characterized using chromatography, electrophoresis after fluorescent labeling of cysteine residues, and tandem mass spectrometry. Functional complementation assays in Saccharomyces cerevisiae were used to obtain and characterize cDNA sequences. Zn-speciation analysis showed that nearly 80% of the Zn extracted from the sporocarps was associated with cysteine-containing peptides in a 5 kDa complex. Screening of an R. atropurpurea cDNA library for sequences encoding peptides capable of sequestering divalent heavy metals was conducted in the Cd-hypersensitive ycf1Δ yeast. This allowed identification of two cDNAs, RaZBP1 and RaZBP2, which protected the metal-sensitive yeast mutants against Cd and Zn, but not Co, Mn or Cu, toxicity. The corresponding RaZBP1 and RaZBP2 peptides consisting of 53 amino acid (AA) residues and sharing 77% identity showed only a limited sequence similarity to known MTs, particularly due to the absence of multiple Cys-AA-Cys motifs. Both RaZBPs were detected in a native Zn-complex of R. atropurpurea and the recombinant RaZBP1 was found associated with Zn and Cd in yeasts. Altogether, the results point to an important role of RaZBPs in the handling of a substantial portion of the Zn pool in R. atropurpurea.