Phytoplankton of Lake Peipsi-Pihkva: species composition, biomass and seasonal dynamics

With 33 years of phytoplankton quantitative studies carried out, a series of qualitative data with a length of over 80 years is at our disposal. About 500 algal species have been found in plankton by different researchers. In different seasons and years 35 main species (dominants and subdominants) form 68–96 % of biomass in L. Pihkva (southern, more eutrophic part) and 60–97 % in L. Peipsi (northern, less eutrophic part). L. Lämmijärv, connecting the two parts is similar to L. Pihkva in respect to phytoplankton and the trophic state. Diatoms and blue-green algae prevail in biomass, diatoms and green algae, in the species number. The oligo-mesotrophic Aulacoseira islandica (O. Müller) Sim. is characteristic of the cool period; A. granulata (Ehr.) Sim. and Stephanodiscus binderanus (Kütz.) Krieger prevail in summer and autumn, the latter being most abundant in the southern part. Gloeotrichia echinulata (J.S. Smith) P Richter and Aphanizomenon flos-aquae (L.) Ralfs dominate in summer causing water-bloom. Phytoplankton has mostly three maxima in seasonal dynamics in L. Peipsi and two in L. Pihkva. Its average biomass in spring in different years has fluctuated in the range 5.6–16 and 6–12.7 g m^–3, in summer 3.1–14.8 and 5.6–125 (10–20 in most cases); and in autumn 7–16.3 and 5.2–26 in the northern and southern parts, respectively.The dominant complex has not changed considerably since 1909; however, the distribution of dominant species in lake parts has become more even in the last decades. Periods of high biomass occurred in the first half of the 1960s and 1970s and in 1988–1994, of low biomass in 1981–1987. The first coincided, in general, with periods of low water level and high water temperature.     

Factors affecting cold hardiness in the small striped flea beetle, Phyllotreta undulata

The striped flea beetle, Phyllotreta undulata Kutschera (Coleoptera: Chrysomelidae), is a pest of cruciferous crops. It overwinters as an adult. During winter in northern European countries, such as Estonia, it is subject to sometimes severe temperatures that may fluctuate daily, over the season, and between seasons. The objective of this study was to investigate factors that affect its cold hardiness. In a series of five experiments, the effects of food plant, starvation, and acclimatization on the beetles’ ability to supercool and survive exposure to sub‐zero temperatures was investigated. The supercooling points (SCP) of overwintered beetles field‐collected from white mustard and Indian mustard differed from those caught from white cabbage and oilseed rape, but these differences disappeared after a 4‐day period of starvation at room temperature, indicating that gut content probably influences the potential to supercool. The duration and temperature of acclimation affected SCP in overwintered beetles. The decrease in SCP was more rapid at 22 °C than at 0 °C, probably because of faster dehydration and gut evacuation at the higher temperature. Acclimation at 0 °C for a week increased the ability of overwintered beetles to survive sub‐zero temperatures, lowering both SCP and lower lethal temperature (LLT₅₀). Some pre‐freeze mortality occurred; SCP and LLT₅₀ were correlated but the latter was a constant 3 °C higher than the former. The SCP of field‐collected pre‐winter beetles decreased gradually during the autumn. It also decreased when field‐collected pre‐winter beetles were acclimated at 0 °C in the laboratory, attaining its lowest level after 18 days. Phyllotreta undulata is well‐adapted to unstable and sometimes severe winter conditions; its high potential to supercool enhances its cold hardiness and ability to survive short periods at sub‐zero temperatures although it cannot survive freezing of its body fluids.     

Purification and characterization of the recombinant human prostaglandin H synthase-2 expressed in Pichia pastoris

Prostaglandin H synthase-1 and -2 (PGHS-1 and PGHS-2, EC 1.14.99.1) are membrane associated glycoproteins that catalyze the first two steps in prostaglandin synthesis. As the enzymes play an important regulatory role in several physiological and pathophysiological processes, recombinant PGHS isoforms are widely used in biomedical research. In the present study, we expressed human PGHS-2 (hPGHS-2) with and without a six histidine sequence tag (His(6) tag) near the amino- or carboxy-terminus of the protein in the Pichia pastoris (P. pastoris) expression system using native or yeast signal sequences. The recombinant His(6) tagged hPGHS-2 was purified using Ni-affinity and anion exchange chromatography, whereas the purification of the C-terminally His(6) tagged hPGHS-2 was more efficient. K(m), k(cat) and IC(50) values were determined to characterize the protein. The data obtained indicate that both the N- and C-terminally His(6) tagged hPGHS-2 are functional and the catalytic properties of the recombinant protein and the enzyme produced in other expression systems are comparable. As the yeast culture is easy to handle, the P. pastoris system could serve as an alternative to the most commonly used baculovirus-insect cell expression system for the production of the recombinant PGHS-2.     

Phytoplankton response to changed nutrient level in Lake Peipsi (Estonia) in 1992–2001

To clarify the sediment yield processes following a disturbance by a forest fire in a mountainous catchment, and considering the hydrological and geomorphological processes in the headwater, we measured bedload sediment yield at rainfall events in disturbed and secondary forest catchments in the western part of Japan. The three catchments were under different hydrogeological conditions. The IK, TB and TY catchments were disturbed by forest fires in 2000, 1994, and 1978, respectively. In the IK catchment, although runoff response to rainfall was fastest with high peak flows, the catchment also had the highest base flow. Moreover, the annual sediment yield there was about ten times as high as in the other two catchments, and it was found that there was a steep linear curve in the relationship between precipitation and bedload sediment yield. This is thought to be caused by overland flow generation following water repellency on the slopes, and by the accumulated sediment that forms the thick soil layer on the valley bottom. On the other hand, in the TB catchment runoff experienced high peak flows at rainfall events and low base flows, and there was a gradual linear curve in the precipitation–sediment yield relationship. This might be the result of there being a thin soil layer on the hillslope and on the valley bottom because of successive erosion after the fire. In the TY catchment, runoff had a low peak flow at rainfall events and a high base flow; and the bedload sediment yield increased exponentially with increasing precipitation. Therefore, sediment yield in the TB catchment was more than that in the TY during storm events with precipitation of less than 100 mm, whereas it was the opposite during heavier rainfalls. It indicates that there is a thick soil layer on the slope and a thin soil layer on the valley bottom in the TY catchment following the recovering of vegetation, and that the sediment yield process predominates only during big rainfall events, only then does subsurface flow generate.     

Preclinical Assessment of the Treatment of Second-Stage African Trypanosomiasis with Cordycepin and Deoxycoformycin

Background

There is an urgent need to substitute the highly toxic compounds still in use for treatment of the encephalitic stage of human African trypanosomiasis (HAT). We here assessed the treatment with the doublet cordycepin and the deaminase inhibitor deoxycoformycin for this stage of infection with Trypanosoma brucei (T.b.).

Methodology/Principal Findings

Cordycepin was selected as the most efficient drug from a direct parasite viability screening of a compound library of nucleoside analogues. The minimal number of doses and concentrations of the drugs effective for treatment of T.b. brucei infections in mice were determined. Oral, intraperitoneal or subcutaneous administrations of the compounds were successful for treatment. The doublet was effective for treatment of late stage experimental infections with human pathogenic T.b. rhodesiense and T.b. gambiense isolates. Late stage infection treatment diminished the levels of inflammatory cytokines in brains of infected mice. Incubation with cordycepin resulted in programmed cell death followed by secondary necrosis of the parasites. T.b. brucei strains developed resistance to cordycepin after culture with increasing concentrations of the compound. However, cordycepin-resistant parasites showed diminished virulence and were not cross-resistant to other drugs used for treatment of HAT, i.e. pentamidine, suramin and melarsoprol. Although resistant parasites were mutated in the gene coding for P2 nucleoside adenosine transporter, P2 knockout trypanosomes showed no altered resistance to cordycepin, indicating that absence of the P2 transporter is not sufficient to render the trypanosomes resistant to the drug.

Conclusions/Significance

Altogether, our data strongly support testing of treatment with a combination of cordycepin and deoxycoformycin as an alternative for treatment of second-stage and/or melarsoprol-resistant HAT.     

The Composition and Density of Epiphyton on Some Macrophyte Species in the Partly Meromictic Lake Verevi

In this paper, some important problems related to taxonomic resolution in water quality assessment by means of macroinvertebrates are discussed. Most quality indices based on macroinvertebrates only require identification up to genus or family level. Although this can be seen as a practical trade-off between taxonomic precision and time constraints and financial resources, it can result in biased assessment scores for certain stream types. An additional difficulty of identification levels other than species is caused by possible changes in taxonomy over time. A given genus may indeed have been split up into two or more genera or a species could be assigned to a different genus. These changes may alter biotic index values calculated over time, due to a change in number of taxa or replacement of one taxon by another one having a different tolerance class. An additional problem is caused by the invasion of exotic species. The genus Corbicula for instance is currently invading Belgian watercourses in increasing numbers. Since no Belgian Biotic Index (BBI) tolerance class is defined for Corbicula, this may cause inconsistencies in index calculations as well. In order to eliminate these, a semi-fixed taxa list, including a tolerance class for each taxon, for BBI calculation is proposed.     

Analysis of changes over 44 years in the phytoplankton of Lake Võrtsjärv (Estonia): the effect of nutrients,climate and the investigator on phytoplankton-based water quality indices

We analysed long-term changes in phytoplankton composition in relation to hydrological, meteorological and nutrient loading data in the large (270 km²) shallow (mean depth 2.8 m) Lake Võrtsjärv. Nutrient loads to the lake were heavy in the 1970s and 1980s and decreased considerably thereafter. The average nutrient concentrations for 1985–2004 (1.6 mg l^?1 of total nitrogen and 53 μg l^?1 of total phosphorus) characterize the lake as a eutrophic water body. All four calculated taxonomic indices showed a unidirectional deterioration of the lake’s ecological status, despite reduced concentrations of nutrients. We focused our analysis on the PTSI index, which revealed a stepwise change between the years 1977 and 1979 that coincided with a large increase in water level, but also with a change of investigator. After correcting input data for possible investigator-induced differences, the step change remained because it was caused by major changes in the whole phytoplankton community. The previous dominant Planktolyngbya limnetica was replaced by two species of seasonally altering Limnothrix. Among phytoplankton functional groups, there was a decrease in all groups comprising small-sized phytoplankton species, such as X1, E, F, J, N and an increase in S1 and H1, both represented by filamentous cyanobacteria. Our results suggest a non-linear response of phytoplankton to changing nutrient loadings, and that the change observed between 1977 and 1979 was a regime shift triggered by water level change. High shade tolerance of the new dominants, and their ability to create shade, obviously stabilized the new status making it resistant to restoration efforts.     

Identification and characterization of an arachidonate 11R-lipoxygenase

11R-Lipoxygenase (11R-LOX) activity has been detected in several marine invertebrates, and here we report the first cloning and expression of the enzyme. The cDNA encoding a protein of 77kDa was isolated by RT-PCR from the soft coral Gersemia fruticosa and expressed in Escherichia coli. Incubations of recombinant enzyme with arachidonic acid yielded a single product, identified by RP-HPLC, GC-MS, and chiral phase-HPLC as 11R-hydroperoxyeicosatetraenoic acid. Other C18, C20, and C22 substrates are also oxygenated, preferentially at the omega10 position. Significantly, both Ca(2+)-ions and a membrane fraction are required for catalytic activity. Calcium effects translocation of the soluble 11R-LOX to the membrane and this association is reversible by Ca(2+) chelation. The enzyme sequence contains some conserved amino acids implicated in calcium activation of mammalian 5-LOX, and with its obligate requirement for membrane interaction the 11R-LOX may thus provide a new model for further analysis of this aspect of lipoxygenase activation.