Detection of free and covalently bound microcystins in animal tissues by liquid chromatography-tandem mass spectrometry

Microcystins are cyanobacterial hepatotoxins capable of accumulation into animal tissues. The toxins act by inhibiting specific protein phosphatases and both non-covalent and covalent interactions occur. The 2-methyl-3-methoxy-4-phenylbutyric acid (MMPB) method determines the total, i.e. the sum of free and protein-bound microcystin in tissues. The aim of the method development in this paper was to tackle the problems with the MMPB methodology: the rather laborious workflow and the loss of material during different steps of the method. In the optimised workflow the oxidation recovery was of acceptable level (29-40%), the extraction efficiency good (62-97%), but the signal suppression effect from the matrix remained severe in our system (16-37% signal left). The extraction efficiency for the determination of the free, extractable microcystins, was found to be good, 52-100%, depending on the sample and the toxin variant and concentration.     

Antioxidant response versus selenium accumulation in the liver and kidney of the Siberian sturgeon (Acipenser baeri)

The aim of this study was to examine the effects of selenium on concentrations of metabolites and enzyme activities acting as antioxidant markers in liver and kidney of Siberian sturgeon Acipenser baeri. Sturgeons were fed selenium cysteine for 30 and 60 d at 1.25, 5, 20 mg Se kg⁻¹. Selenium level in the control feed was 0.32 mg kg⁻¹. Se concentration was measured in liver, kidney and muscle of every specimen. Sturgeon accumulated Se in tissues with a clear dose-response relationship and the highest Se concentration was recorded in liver. This outcome is lined up with the findings obtained on the antioxidant markers evaluated in both tissues, and in which a dose-response for several biomarkers was recorded in liver. The superoxide dismutase activity in Se-treated fish was generally induced, while catalase activity was lower in liver or unaltered in kidney. The concentrations of glutathione S-transferase, glutathione reductase and total glutathione responded differently for both tissues and were induced in a different way at both endpoints. No changes of glyoxalase I activity were noted for both Se-treated tissues, while for glyoxalase II enzyme in liver a dose-related pattern was found showing a reversible effect (decreased and increased counteractive response) only in the 5 mg kg⁻¹ group. Moreover, the highest Se concentrations did not cause marked changes in malondialdehyde levels of liver and kidney. The enhancement of glutathione peroxidase activity in Se-treated sturgeon might have prevented the lipid peroxidation in both tissues, providing to the Siberian sturgeon a great defense ability versus the prooxidant effect of selenium.     

Characterization of cadmium (Cd) distribution and accumulation in Tagetes erecta L. seedlings: Effect of split-root and of remove-xylem/phloem

Tagetes erecta has a high potential for cadmium (Cd) phytoremediation. Through several hydroponic experiments, characteristics of ¹⁰⁸Cd distribution and accumulation were investigated in T. erecta with split -roots or removed xylem/phloem. The results showed that ¹⁰⁸Cd transport from roots to aboveground tissues showed the homolateral transport phenomenon in split-root seedlings. ¹⁰⁸Cd content of leaves on the +¹⁰⁸Cd side and the −¹⁰⁸Cd side was not significantly different, which implied that there was horizontal transport of ¹⁰⁸Cd from the +¹⁰⁸Cd side to the −¹⁰⁸Cd side in cut-root seedlings. Like ¹⁰⁸Cd transport, the transport of ⁷⁰Zn was homolateral. Reduction of water consumption in the removed xylem treatment significantly decreased ¹⁰⁸Cd accumulation; whereas, the removed phloem treatment had no significant effect on water consumption, but did decrease ¹⁰⁸Cd accumulation in leaves of the seedlings. The removal of phloem significantly reduced distal leaf ¹⁰⁸Cd content, which was significantly lower than that in the basal leaves in both the split-root and unsplit-root seedlings. Overall, the results presented in this study revealed that the root to aboveground cadmium translocation via phloem is as an important and common physiological process as xylem determination of the cadmium accumulation in stems and leaves of marigold seedlings.     

Effects of different inorganic arsenic species in Cyprinus carpio (Cyprinidae) tissues after short-time exposure: Bioaccumulation, biotransformation and biological responses

Differences in the toxicological and metabolic pathway of inorganic arsenic compounds are largely unknown for aquatic species. In the present study the effects of short-time and acute exposure to As^III and As^V were investigated in gills and liver of the common carp, Cyprinus carpio (Cyprinidae), measuring accumulation and chemical speciation of arsenic, and the activity of glutathione-S-transferase omega (GST Ω), the rate limiting enzyme in biotransformation of inorganic arsenic. Oxidative biomarkers included antioxidant defenses (total glutathione-S-transferases, glutathione reductase, glutathione, and glucose-6-phosphate dehydrogenase), total scavenging capacity toward peroxyl radicals, reactive oxygen species (ROS) measurement and lipid peroxidation products. A marked accumulation of arsenic was observed only in gills of carps exposed to 1000 ppb As^V. Also in gills, antioxidant responses were mostly modulated through a significant induction of glucose-6-phosphate dehydrogenase activity which probably contributed to reduce ROS formation; however this increase was not sufficient to prevent lipid peroxidation. No changes in metal content were measured in liver of exposed carps, characterized by lower activity of GST Ω compared to gills. On the other hand, glutathione metabolism was more sensitive in liver tissue, where a significant inhibition of glutathione reductase was concomitant with increased levels of glutathione and higher total antioxidant capacity toward peroxyl radicals, thus preventing lipid peroxidation and ROS production. The overall results of this study indicated that exposure of C. carpio to As^III and As^V can induce different responses in gills and liver of this aquatic organism.     

Larval performance of the mustard leaf beetle (Phaedon cochleariae, Coleoptera, Chrysomelidae) on white mustard (Sinapis alba) and watercress (Nasturtium officinale) leaves in dependence of plant exposure to ultraviolet radiation

Short-term exposure to ambient or attenuated ultraviolet (UV) radiation resulted in shifts in plant metabolite concentrations of the Brassicaceae Sinapis alba and Nasturtium officinale. Leaf quality also varied between plant species and within species due to age. Larvae of the oligophagous leaf beetle Phaedon cochleariae were raised on these different host leaves, in order to investigate the effects of variable plant chemistry on this herbivore. The performance of P. cochleariae was influenced by chemical differences between and within plant species but it responded with high plasticity to plants stressed by ultraviolet radiation. Body mass increase and developmental times of larvae were exclusively affected by plant species and leaf-age. However, developmental differences were fully compensated in the pupal stage. We suggest that the plasticity of herbivores may depend on the degree of specialisation, and insect performance may not necessarily be altered by stress-induced host plants.     

Accumulation features and temporal trends of PCDDs, PCDFs and PCBs in Baikal seals (Pusa sibirica)

This study investigated the accumulation features and temporal trends of PCDD/Fs, dioxin-like PCBs (DL-PCBs) and non-dioxin-like PCBs (NDL-PCBs) in the blubber of Baikal seals collected in 1992 and 2005. DL-PCBs (480-3600 ng/g) and NDL-PCBs (980-35,000 ng/g) were dominant contaminants. Concentrations of PCDDs and PCBs in males were significantly higher than in females. In males, age-dependent accumulation was observed for PCDDs, mono-ortho PCBs and NDL-PCBs. PCDFs and non-ortho PCBs showed no such trends, implying that exposure of seals to these contaminants has been decreasing in recent years. No decreasing temporal trend was observed for PCDDs, mono-ortho PCBs and NDL-PCBs, suggesting that Baikal seals are still exposed to PCDDs and PCBs. TEQs of PCDDs and mono-ortho PCBs in seals collected in 2005 accounted for 62-77% of total TEQs. The TEQ levels in 40% of the specimens exceeded the threshold level for immunosuppression observed in harbor seals (209 pg/g).     

Lead exposure and biological effects in pied flycatchers (Ficedula hypoleuca) before and after the closure of a lead mine in northern Sweden

Mining activities affect the surrounding environment by increasing exposure to metals. In this study, metal accumulation and its effects on reproduction and health of pied flycatcher (Ficedula hypoleuca) nestlings were monitored before and up to five years after a lead mine and enrichment plant closed down. The lead concentration in moss, nestling blood, liver and feces all indicated decreased lead exposure by at least 31% after closure, although only blood lead decreased significantly. Although the birds responded fairly well to the changed atmospheric deposition (based on moss samples), concentrations were still higher compared with birds in a reference area, and breeding was affected at the mine (smaller clutches and higher mortality). Surviving nestlings suffered from lower hemoglobin levels, mean cell hemoglobin concentrations and inhibited δ-aminolevulinic acid dehydratase activity. Lead poisoning contributed to poor health and adverse reproductive effects, but other factors (e.g. increased parasitic load) probably also affected the birds.