Multivariate Statistics of the Pyrolysis Products of High Molecular Components in Pulping Wastewaters to Explain Their Toxicity (6 pp)

Background, Aim and Scope At present, large-scale paper manufacture involves delignification and bleaching by elemental chlorine free (ECF) or totally chlorine free (TCF) processes. The wastewater is purified by secondary treatment (mechanical, chemical and biological) which removes most of the toxic substances from the discharge. However, we found residual toxicity in the high molecular (> 1000 D) matter (HMWM) of the discharge by test of the RET (reverse electronic transfer) inhibition. This fraction consists mainly of polydisperse lignin (LIG) and carbohydrate (CH) macromolecules. Structural units in these molecules are studied by pyrolysis gas chromatography / mass spectrometry (Pyr-GC/MS). In the present work, our aim was to find out those structural units which could explain the RET toxicity of LIG or CH molecules. We compared statistically RET toxicity values of the HMWM samples from treated wastewaters of pilot pulping experiments and intensity variation of the pyrolysis product gas chromatograms of these samples. This application is a novel study procedure. Methods Pyrolysis products (Py-GC/MS results) and inhibition of RET (reverse electronic transport toxicity) as TU50 and TU20 of HMWM (High Molecular Weight Material; Mw > 1000 D) were compared by multivariate statistics. The samples were from laboratory pilot stages of TCF (Totally Chlorine Free) and ECF (Elemental Chlorine Free) manufacture of softwood pulp. Py-GC/MS was done without and with addition of TMAH (Tetra Methyl Ammonium Hydroxide). The name and structure of each abundant fragment compound was identified from its retention time and mass spectrum compared to authentic reference compounds or literature. Four sets of Toxicity Units (TUs) and GC peak areas of the pyrolysis fragments were obtained. The data were normalized by division with LIG (lignin content of each sample). TU values were dependent and the fragment values independent (explanatory) variables in statistical treatments by SPSS system. Separate analyses of correlations, principal components (PCA) and stepwise multiple linear regression (SMLR) were performed from the four sample sets TCF and ECF with and without TMAH. Results and Discussion From the CH fragments, 2-furfural in TCF, and from the LIG fragments, styrene in ECF showed the highest probabilities to originate from source structures of toxicity. Other possible compounds in concern were indicated to be CH fragment 2-methyl-2-cyclopenten-1-one in ECF and LIG fragments 2-methoxy-4-methylphenol, 4,5-dimethoxy-2-methylphenol and 2-methylphenol in TCF.     

Polychlorinated dibenzothlophenes in bleached pulp mill effluents

Polychlorinated dibenzothiophenes, PCDBTs, have been analysed with HRGC/HRMS from different bleached pulp mill effluents. 2,3,7,8-TeCDBT was found to be the dominating TeCDBT isomer in some samples. The estimated concentrations of the TeCDBTs in the samples were < 1 to 60 pg/L. PeCDBTs and other higher chlorinated PCDBTs were not found (< 1 pg/L).     

Biocides in eggs of aquatic birds. Completion of a food chain enrichment study for DDT,PCB and HG

Eggs (N=224) of seven aquatic bird species of Lake Päijänne, Finland, were analyzed for their contents of HCB, DDE, PCB, mercury (TOTHG) and methyl mercury (METHG as per cent of TOTHG). METHG was 85.5 % which is the same as in fish. Also the levels of TOTHG were of the same order of magnitude in eggs and in fish of Päijänne, but the levels of chlorinated hydrocarbons were much higher in eggs. Differences between species were highly significant but the annual and regional differences were mostly insignificant. The chlorinated hydrocarbons correlated positively with each other and with fat contents but insignificantly with mercury contents. Calculation of total residue amounts in egg, juvenile and adult bird specimens exposed an enrichment of DDT to three, PCB to five and mercury to all seven species.     

A wood preservative metabolite in river water

- A previously unknown pollutant in river water was identified to be 2-mercaptobenzothiazole (2-MBT) by interpretation and simulation of its GC/LRMS spectrum. Further GC/HRMS measurement of the isotope composition of the molecular ion verified this structure. 2-MBT is a well-known agent for corrosion inhibition and a stable metabolite of several other benzothiazoles. The present 2-MBT trace was most probably a metabolite of the wood preservative TCMTB which leaked from an upstream sawmill. The metabolite had been detected earlier in urine of the sawmill workers, but now was identified in the recipient water environment for the first time.     

Finnish Lake Survey: The Role of Catchment Attributes in Determining Nitrogen, Phosphorus, and Organic Carbon Concentrations

This study is based on a Finnish lake survey conducted in 1995, a dataset of 874 statistically selected lakes from the national lake register. The dataset was divided into subgroups to evaluate lake water-catchment relationships in different geographical regions and in lakes of different size. In the three southernmost regions, the coefficients of determination in multiple regression equations varied between 0.40 and 0.53 for total nitrogen (TN) and between 0.37 and 0.53 for total phosphorus (TP); the best interpreters were agricultural land and water area in the catchment. In the two northernmost regions, TN concentrations in lake water were best predicted by the proportion of peatlands in the catchment, the catchment slope, and TP concentrations by lake elevation and latitude. Coefficients of determination in multiple regression equations in these northern regions varied between 0.39 and 0.67 for TN and between 0.41 and 0.52 for TP. For all the subsets formed, the best coefficients of determination explaining TN, TP, and total organic carbon (TOC) were obtained for a subset of large lakes (>10 km²), in which 72–83% of the variation was explained. This was probably due to large heterogeneous catchments of these lakes.     

Biological responses of midge (Chironomus riparius) and lamprey (Lampetra fluviatilis) larvae in ecotoxicity assessment of PCDD/F-, PCB- and Hg-contaminated river sediments

Environmental Science and Pollution Research - We evaluated the utility of chironomid and lamprey larval responses in ecotoxicity assessment of polychlorinated dibenzo-p-dioxins, dibenzofurans...     

Changes in Lumbriculus variegatus metabolites under hypoxic exposure to benzo(a)pyrene,chlorpyrifos and pentachlorophenol: Consequences on biotransformation

The regulation of endogenous metabolites is still not fully understood in aquatic invertebrates exposed concurrently to toxicants and hypoxia. Despite the prevalence of hypoxia in the aquatic environment, toxicity estimations seldom account for multiple stressors thereby differing from natural conditions. In this study, we examined the influence of hypoxia (<30% O₂) on contaminant uptake and the composition of intracellular metabolites in Lumbriculus variegatus exposed to benzo(a)pyrene (B(a)P, 3 μg L⁻¹), chlorpyrifos (CPF, 100 μg L⁻¹) or pentachlorophenol (PCP, 100 μg L⁻¹). Tissue extracts of worms were analyzed for 123 metabolites by gas chromatography–mass spectrometry and metabolite levels were then related to treatments and exposure time. Hypoxia markedly increased the accumulation of B(a)P and CPF, which underlines the significance of oxygen in chemical uptake. The oxygen effect on PCP uptake was less pronounced. Succinate and glycerol-3-phosphate increased significantly (p < 0.0001) following hypoxic treatment, whereas sugars, cysteine, and cholesterol were effectively repressed. The buildup of succinate coupled with the corresponding decline in intracellular 2-oxo- and 2-hydroxy glutaric acid is indicative of an active hypoxia inducible factor mechanism. Glutamate, and TCA cycle intermediates (fumarate, and malate) were disturbed and evident in their marked suppression in worms exposed concurrently to hypoxia and PCP. Clearly, hypoxia was the dominant stressor for individuals exposed to B(a)P or CPF, but to a lesser extent upon PCP treatment. And since oxygen deprivation promotes the accumulation of different toxicants, there may be consequences on species composition of metabolites in natural conditions.     

Gene expression responses of paper birch (Betula papyrifera) to elevated CO2 and O3 during leaf maturation and senescence

Gene expression responses of paper birch (Betula papyrifera) leaves to elevated concentrations of CO₂ and O₃ were studied with microarray analyses from three time points during the summer of 2004 at Aspen FACE. Microarray data were analyzed with clustering techniques, self-organizing maps, K-means clustering and Sammon's mappings, to detect similar gene expression patterns within sampling times and treatments. Most of the alterations in gene expression were caused by O₃, alone or in combination with CO₂. O₃ induced defensive reactions to oxidative stress and earlier leaf senescence, seen as decreased expression of photosynthesis- and carbon fixation-related genes, and increased expression of senescence-associated genes. The effects of elevated CO₂ reflected surplus of carbon that was directed to synthesis of secondary compounds. The combined CO₂ + O₃ treatment resulted in differential gene expression than with individual gas treatments or in changes similar to O₃ treatment, indicating that CO₂ cannot totally alleviate the harmful effects of O₃.     

Effects of ultraviolet-B radiation and larval growth on toxicokinetics of waterborne bisphenol A in common frog (Rana temporaria) larvae

In 1989, researchers discovered that amphibians, particularly frogs and toads from many parts of the world, appeared to be declining. In many ecosystems amphibians play a central role in ecosystem energy flow and nutrient cycling, and they act as keystone species. The recent increase in solar ultraviolet-B radiation (UVB, 280-320nm) has been thought to be one stressor responsible for the decline in amphibian populations. Along with other stressors, such as habitat destruction, anthropogenic influences and natural causes, UVB radiation could contribute to adverse effects among amphibians. Amphibians provide a good model for examining the effects of environmental stressors, because both lethal and sub-lethal responses are well documented in a range of studied xenobiotics in many species. In this experiment, the effects of UVB radiation on the accumulation and depuration kinetics of bisphenol A (BPA) were studied. Additionally, the accumulation was further modeled with correction for growth dilution. The results indicate that UVB radiation did not affect the toxicokinetics of BPA, and that the applied growth correction had only a negligible influence on the toxicokinetic estimations in this experiment. However, BCFs values calculated as k(u)/k(e) where closer to C(a)/C(w) calculated values when growth dilution was incorporated in the model. This method can be used in other experiments, where the growth dilution can affect toxicokinetic estimations.     

Extensive tree health monitoring networks are useful in revealing the impacts of widespread biotic damage in boreal forests

We surveyed the regional distribution of conifer defoliation in Finland with an extensive monitoring network during 1995–2006 (EU Forest Focus Level I). The average defoliation in the whole Finland was 10.3% in pine and 19.9% in spruce. The sharp changes were often related to abiotic and biotic factors. The mean age of the stand explained more than one half of the between-plot variance in defoliation. In a variance component analysis, the main effect of years was negligible, while most of the random variation was due to plot main effect and plot × year interaction. About one fifth of the defoliation could be attributed to abiotic or biotic damage, and there were strong local correlations, e.g., between the changes in defoliation and degree of pine sawfly (Diprionidae) damage. There were clear temporal and spatial patterns in the incidence of the most important causes [Scots pine: Scleroderris canker (Gremmeniella abietina), pine shoot beetles (Tomicus sp.), and pine sawflies (Diprion pini, Neodiprion sertifer); Norway spruce: rust fungi (primarily Chrysomyxa ledi)]. Our results suggest that extensive monitoring networks can reveal useful information about the widespread outbreaks of pest organisms (insects and fungi) already in their increase phases, giving some time for management decisions. In a changing climate, large-scale, regular monitoring of tree health, including abiotic and biotic causes, is more important than ever before.