Spawning, egg development and early ontogenesis in rock cod Patagonotothen ramsayi (Regan, 1913) caught on the Patagonian Shelf and maintained in captivity

Rock cod Patagonotothen ramsayi (Regan, 1913) is one of the most abundant fish of the family Nototheniidae inhabiting the Patagonian Shelf and upper Slope in the southwest Atlantic. Recently, P. ramsayi became an important commercial species around the Falkland Islands with annual catch of 60,000–75,000 t. The present study aimed to reveal previously unknown aspects of reproductive biology of P. ramsayi during the first successful maintenance of adults for more than a year in an aquaculture facility with running seawater. The fish spawned at the end of austral winter. During spawning, males changed their coloration dramatically, occupied artificial shelters on the bottom and showed aggressive territorial behaviour. Egg masses were light-yellow to light-orange irregular spongiform. They were negatively buoyant, but located outside shelters and were ignored by males. Egg diameters varied between 2.1 and 2.3 mm, and the number of eggs per egg mass ranged from 26,800 to 123,400. Embryogenesis lasted 28–32 days. Total lengths of newly hatched larvae ranged from 6.2 to 6.7 mm. The yolk sac feeding period lasted approximately 11 days, during which the larvae showed negative phototaxis. One-month-old larvae attained 8.8–9.0 mm in length. This study confirms that P. ramsayi exhibit the reproductive strategy typical for nototheniid species occupying low-latitude peripheries of their distributional range, characterised by a combination of r-features (small eggs and larvae, high fecundity) and K-features (territorial behaviour and possible nest guarding).     

Initiation of poliovirus negative-strand RNA synthesis requires precursor forms of p2 proteins

The replication proteins encoded in the P2 region of the poliovirus genome induce extensive rearrangement of cellular membranes into vesicles and are a required component of viral RNA replication complexes. To identify distinct viral protein(s) from the P2 region of the genome that were required to form functional RNA replication complexes, the P2 proteins were expressed in addition to P3 in HeLa S10 translation-RNA replication reactions. Membrane-associated preinitiation replication complexes were isolated from these reactions and used to measure negative-strand synthesis. The formation of replication complexes capable of initiating negative-strand synthesis was observed when either P23 or when P2 and P3 were expressed in the HeLa S10 translation-replication reactions. The amount of negative-strand RNA synthesized with P2 and P3 was approximately 50% of that observed with P23. Negative-strand synthesis was not observed when the processed forms of the P2 proteins (e.g., 2A, 2B, 2C, 2AB, and 2BC) were used in various combinations in place of P2. In contrast, the expression of 2A and 2BCP3 supported negative-strand synthesis at the same level observed with P23. Therefore, functional replication complexes were formed in reaction mixtures that contained either 2A and 2BCP3 or P2 and P3. Genetic complementation analysis of P23 RNA that contained a lethal mutation in 2C confirmed these results. The expression of 2BCP3 in trans restored the replication of P23-2C(P131N) RNA to wild-type levels. The expression of P2 and P3 also complemented the replication of this mutant RNA, although very inefficiently. Complementation was not observed in reactions that contained P2 alone, 2BC, or 2C. Based on these results, we propose that RNA replication complexes are initially formed with the primary cleavage products of P23 (i.e., P2 and P3 or 2A and 2BCP3), and that 2A and 2BCP3 are preferentially used in this process.     

Methanogenic communities in permafrost-affected soils of the Laptev Sea coast, Siberian Arctic, characterized by 16S rRNA gene fingerprints

Permafrost environments in the Arctic are characterized by extreme environmental conditions that demand a specific resistance from microorganisms to enable them to survive. In order to understand the carbon dynamics in the climate-sensitive Arctic permafrost environments, the activity and diversity of methanogenic communities were studied in three different permafrost soils of the Siberian Laptev Sea coast. The effect of temperature and the availability of methanogenic substrates on CH4 production was analysed. In addition, the diversity of methanogens was analysed by PCR with specific methanogenic primers and by denaturing gradient gel electrophoresis (DGGE) followed by sequencing of DGGE bands reamplified from the gel. Our results demonstrated methanogenesis with a distinct vertical profile in each investigated permafrost soil. The soils on Samoylov Island showed at least two optima of CH4 production activity, which indicated a shift in the methanogenic community from mesophilic to psychrotolerant methanogens with increasing soil depth. Furthermore, it was shown that CH4 production in permafrost soils is substrate-limited, although these soils are characterized by the accumulation of organic matter. Sequence analyses revealed a distinct diversity of methanogenic archaea affiliated to Methanomicrobiaceae, Methanosarcinaceae and Methanosaetaceae. However, a relationship between the activity and diversity of methanogens in permafrost soils could not be shown.     

Dynamic Histone Acetylation of Late Embryonic Genes during Seed Germination

Histone acetylation is involved in the regulation of gene expression in plants and eukaryotes. Histone deacetylases (HDACs) are enzymes that catalyze the removal of acetyl groups from histones, which is associated with the repression of gene expression. To study the role of histone acetylation in the regulation of gene expression during seed germination, trichostatin A (TSA), a specific inhibitor of histone deacetylase, was used to treat imbibing Arabidopsis thaliana seeds. GeneChip arrays were used to show that TSA induces up-regulation of 45 genes and down-regulation of 27 genes during seed germination. Eight TSA-up-regulated genes were selected for further analysis – RAB18, RD29B, ATEM1, HSP70 and four late embryogenesis abundant protein genes (LEA). A gene expression time course shows that these eight genes are expressed at high levels in the dry seed and repressed upon seed imbibition at an exponential rate. In the presence of TSA, the onset of repression of the eight genes is not affected but the final level of repressed expression is elevated. Chromatin immunoprecipitation and HDAC assays show that there is a transient histone deacetylation event during seed germination at 1 day after imbibition, which serves as a key developmental signal that affects the repression of the eight genes. Electronic supplementary material Electronic supplementary material is available for this article at and accessible for authorised users.     

Chloraminated drinking water does not generate bacterial resistance to antibiotics in Pseudomonas aeruginosa biofilms

Aim: To determine if exposure of Pseudomonas aeruginosa biofilms to chloraminated drinking water can lead to individual bacteria with resistance to antibiotics. Methods and Results: Biofilms of P. aeruginosa PA14 were grown in drinking water in a Kadouri drip‐fed reactor; the biofilms were treated with either 0·5 mg l^‐1 or 1·0 mg l^‐1 of chloramine for 15 or 21 days; control biofilms were grown in water without chloramine. Fewer isolates with antibiotic resistance were obtained from the chloramine‐treated biofilms as compared to the control. Minimum inhibitory concentrations (MIC) for selected antibiotic‐resistant isolates were determined using ciprofloxacin, tobramycin, gentamicin, rifampicin and chloramphenicol. All of the isolates tested had increased resistance over the wildtype to ciprofloxacin, rifampicin and chloramphenicol, but were not resistant to tobramycin or gentamicin. Conclusions: Under these test conditions, there was no detectable increase in antibiotic resistance in P. aeruginosa exposed as biofilms to disinfectant residues in chloraminated drinking water. Significance and Impact of the study: Chloramine in drinking water, while unable to kill biofilm bacteria, does not increase the potential of P. aeruginosa to become resistant to antibiotics.     

Total RNA isolation from<Emphasis Type="Italic">Picea mariana</Emphasis> dry seed

Isolating RNA from dry conifer seeds can be difficult because of a number of interfering compounds present in seeds. We describe a protocol for total RNA isolation from black spruce dry seeds, which is an adaptation of a method used for mouse myeloma tissues. The extraction relies on selective precipitation of RNA by using lithium chloride.     

Variability in rate of human perspiration in european populations

1. 1. In the consideration of ergonomic tasks the attempt is often made to combine the results of physiological research on human perspiration with technical factors such as the transfer of moisture through specific materials.

2. 2. The result of such attempts is often unsatisfactory due to the fact that the percentiles and frequency distribution of perspiration rates are often unknown or not taken into consideration.

3. 3. Further, the interaction between the human user and a particular environmental situation remains unaccounted for.

4. 4. This interaction can produce totally different reactions in both the user and the materials depending upon the specifics of the situation.

5. 5. In response to this deficit we tested 738 persons from various areas in Europe under laboratory conditions to determine perspiration rates and their statistical distribution.

6. 6. Despite the climatic differences existing within Europe, a uniform picture characterized by an extremely skewed distribution resulted.

7. 7. However, it was also shown that these values, including their statistical distribution and percentile ranks, can be strongly influenced by specific external conditions.

8. 8. In the design of human working and living environments it is necessary to examine the situation with research subjects whose perspiration rates are known in order to gain insight into the interactions existing between the human user and the material or technical aspects of the particular environment in question.

Fungal diversity and deterioration in mummified woods from the ad Astra Ice Cap region in the Canadian High Arctic

Non-permineralized or mummified ancient wood found within proglacial soil near the ad Astra Ice Cap (81°N, 76°W), Ellesmere Island, Canada was investigated to ascertain the identification of the trees, current morphological and chemical characteristics of the woods and the fungi within them. These woods, identified as Betula, Larix, Picea and Pinus, were found with varying states of physical and chemical degradation. Modern microbial decomposition caused by soft rot fungi was evident and rDNA sequencing of fungi obtained from the samples revealed several species including Cadophora sp., Exophiala sp., Phialocephala sp., as well as others. Analytical ¹³C-labeled tetramethylammonium hydroxide thermochemolysis showed the lignin from the ancient wood was in a high degree of preservation with minor side chain alteration and little to no demethylation or ring hydroxylation. The exposure of these ancient woods to the young soils, where woody debris is not usually prevalent, provides carbon and nutrients into the polar environment that are captured and utilized by unique decay fungi at this Arctic site.     

Continuous production of isopropanol and butanol using <Emphasis Type="Italic">Clostridium beijerinckii</Emphasis> DSM 6423

Clostridium beijerinckii DSM 6423 was studied using different continuous production methods to give maximum and stable production of isopropanol and n-butanol. In a single-stage continuous culture, when wood pulp was added as a cell holding material, we could increase the solvent productivity from 0.47 to 5.52 g L⁻¹ h⁻¹ with the yield of 54% from glucose. The overall solvent concentration of 7.51 g L⁻¹ (39.4% isopropanol and 60.6% n-butanol) with the maximum solvent productivity of 0.84 g L⁻¹ h⁻¹ was obtained with two-stage continuous culture. We were able to run the process for more than 48 overall retention times without losing the ability to produce solvents.