Functional roles of ants in a temperate grassland.

Ants in temperate grasslands are consumers and ecosystem engineers, influencing biodiversity and potentially grassland productivity. However, the effects of ant exclusion or suppression on resource removal and the biological community in temperate grasslands have yet to be fully explored. We conducted ant-suppression experiments and evaluated the effects of ants on ground-dwelling arthropod communities in the field by using pitfall and bait traps. In the laboratory, we evaluated the effects of ants on the ant-attended aphid Aphis rumicis, which is a honeydew resource for ants, and the slug (Deroceras laeve), an aphid predator. Aboveground arthropod communities were not affected by the ant-suppression treatment. However, slugs (D. laeve and Ambigolimax valentianus) visited bait resources more frequently in the ant-suppression treatment area. In the ant-absence condition in the laboratory experiment, there were fewer aphids on the plants compared to the ant-presence condition owing to predation by D. laeve. Our results suggest that ant abundance in temperate grasslands influences the predation activity of slugs toward honeydew sources such as aphids.

The succession of microbial community in the organic rich fish-farm sediment during bioremediation by introducing artificially mass-cultured colonies of a small polychaete, Capitella sp. I.

We monitored seasonal changes of the abundance and composition of microorganisms in the fish-farm sediment in Kusuura Bay, Amakusa, Japan, using the quinone profiling technique, during bioremediation by introducing cultured colonies of polychaete, Capitella sp. I. In November 2004, approximately 9.2 million cultured worms were transferred to the fish-farm sediment, which increased rapidly, and reached 458.5 gWW/m(2) (528,000 indiv./m(2)) in March 2005. During this fast-increasing period of Capitella, the microbial quinone content of the surface sediment (0-2 cm) also increased markedly, and reached 237 micromol/m(2) in January 2005, although the water temperature decreased to the lowest levels in the year. Particularly, the mole fraction of ubiquinone-10 in total quinones in the sediment, indicating the presence of alpha subclass of Proteobacteria, increased by 9.3%. These facts suggest that the bacterial growth was enhanced markedly by the biological activities of worms in the sediment, and the bacteria played an important role in the decomposition of the organic matter in the sediment.

Microbial responses using denaturing gradient gel electrophoresis to oil and chemical dispersant in enclosed ecosystems.

Microbial responses to the addition of oil with or without a chemical dispersant were examined in mesocosm and microcosm experiments by using denaturing gradient gel electrophoresis of bacterial ribosomal DNA and direct cell counting. When a water-soluble fraction of oil was added to seawater, increases in cell density were observed in the first 24h, followed by a decrease in abundance and a change in bacterial species composition. After addition of an oil-dispersant mixture, increases in cell density and changes in community structure coincided, and the amount of bacteria remained high. These phenomena also occurred in response to addition of only dispersant. Our results suggest that the chemical dispersant may be used as a nutrient source by some bacterial groups and may directly or indirectly prevent the growth of other bacterial groups.

Correlation between Possession of a Respiration-Dependent Na Pump and Na Requirement for Growth of Marine Bacteria.

The possession of a respiration-dependent primary sodium pump and the requirement of Na for growth were investigated in bacterial isolates from marine environments. The bacteria in which NADH oxidase specifically required Na for maximum activity were believed to possess a primary sodium pump. All bacteria that failed to grow without the addition of NaCl possessed a primary Na pump. All bacteria that had no primary Na pump grew without additional NaCl. The primary Na pump seems to be involved in the Na requirement of marine bacteria, and this can be regarded as a criterion for the definition of marine bacteria.

Impacts by heavy-oil spill from the Russian tanker Nakhodka on intertidal ecosystems: recovery of animal community.

The impact of a heavy-oil spill from the Nakhodka on an intertidal animal community, and the recovery process of animals from the damage were surveyed from the autumn of 1997 to the spring of 2001. The field study was carried out in the rocky coast of Imago-Ura Cove, located along the Sea of Japan, where clean-up operations for oil pollution had been conducted less intensely than in other polluted areas. We have examined individual number of each animal taxon by continuously placing a quadrat of 5 m width along the entire intertidal zone of the cove. A total of 76 invertebrate taxa including 57 species of mollusks, 10 species of crustaceans were observed during the survey. The number of taxa increased from 1998 to 1999 in areas where the initial oil pollution was intense. Total individual number of benthic animals continued to increase from 1998 to 2000 in the polluted areas. The impact of oil on benthic animals was different from species to species. Some species such as Cellana toreuma and Monodonta labio confusa increased rapidly after the oil spill, whereas other species such as Patelloida saccharina lanx and Septifer virgatus did not show any apparent temporal tendencies. Population size structure of P. saccharina lanx varied greatly among years, however that of M. labio confusa did not. For P. saccharina lanx, recruitment was unsuccessful in 1997, possibly due to the effect of oil pollution. These differences in responses to oil pollution among benthic animals are considered to be caused by the differences in habitat use, susceptibility to heavy-oil, life history and migration ability. The findings suggest that it took at least 2-3 years for the intertidal animal community to recover to its original level after the oil spill.

Study of the effect of water-soluble fractions of heavy-oil on coastal marine organisms using enclosed ecosystems, mesocosms.

Mesocosm facilities composed of 4 experimental and 2 reservoir tanks (1.5 m in diameter, 3.0 m in depth and 5 tons in capacity) made of FRP plastics, were constructed in the concrete fish rearing pond in the Fisheries Laboratory, The University of Tokyo. The water-soluble fraction of Rank A heavy residual oil was formed by mixing 500 g of the oil with 10 l of seawater, which was introduced to the 5000 l-capacity tanks. Experimental Run 4 was conducted from May 31 to June 7, 2000. Oil concentrations in the tanks were 4.5 microg/l called LOW, and 13.5 microg/l, called HIGH tank. Bacterial growth rates very quickly accelerated in the HIGH tank just after the loading of oil which corresponded with a high increase of bacterial cells in the same tank after 2 days. Later, bacterial numbers in HIGH tank rapidly decreased, corresponding with the rapid increase of heterotrophic nano-flagellates and virus numbers on the same day. Sediment traps were deployed at the bottom of the experimental tanks, and were periodically retrieved. These samples were observed both under light microscope and epi-fluorescent microscope with UV-excitation. It was observed that the main components of the vertical flux were amorphous suspended matter, mostly originating from dead phytoplankton and living diatoms. It was further observed from the pictures that vertical transport of oil emulsions were probably conducted after adsorption to amorphous suspended matter and living diatoms, and were settling in the sediment traps at the bottom of the tanks. This means that the main force which drives the soluble fraction of oil into bottom sediment would be vertical flux of such amorphous suspended particles and phytoplankton. Further incubation of the samples revealed that the oil emulsions were degraded by the activity of autochtonous bacteria in the sediment in aerobic condition.

Study on the fate of petroleum-derived polycyclic aromatic hydrocarbons (PAHs) and the effect of chemical dispersant using an enclosed ecosystem, mesocosm.

Polycyclic Aromatic Hydrocarbons (PAHs) are one of the components found in oil and are of interest because some are toxic. We studied the environmental fate of PAHs and the effects of chemical dispersants using experimental 500 l mesocosm tanks that mimic natural ecosystems. The tanks were filled with seawater spiked with the water-soluble fraction of heavy residual oil. Water samples and settling particles in the tanks were collected periodically and 38 PAH compounds were analyzed by gas chromatography-mass spectrometry (GC-MS). Low molecular weight (LMW) PAHs with less than three benzene rings disappeared rapidly, mostly within 2 days. On the other hand, high molecular weight (HMW) PAHs with more than four benzene rings remained in the water column for a longer time, up to 9 days. Also, significant portions (10-94%) of HMW PAHs settled to the bottom and were caught in the sediment trap. The addition of chemical dispersant accelerated dissolution and biodegradation of PAHs, especially HMW PAHs. The dispersant amplified the amounts of PAHs found in the water column. The amplification was the greater for the more hydrophobic PAHs, with an enrichment factor of up to six times. The increased PAHs resulting from dispersant use overwhelmed the normal degradation and, as a result, higher concentrations of PAHs were observed in water column throughout the experimental period. We conclude that the addition of the dispersant could increase the concentration of water column PAHs and thus increase the exposure and potential toxicity for organisms in the natural environment. By making more hydrocarbon material available to the water column, the application of dispersant reduced the settling of PAHs. For the tank with dispersant, only 6% of chrysene initially introduced was detected in the sediment trap whereas 70% was found in the trap in the tank without dispersant.

Increase in Alphaproteobacteria in association with a polychaete, Capitella sp. I, in the organically enriched sediment.

We conducted bioremediation experiments on the organically enriched sediment on the sea floor just below a fish farm, introducing artificially mass-cultured colonies of deposit-feeding polychaete, Capitella sp. I. To clarify the association between the Capitella and bacteria on the efficient decomposition of the organic matter in the sediment in the experiments, we tried to identify the bacteria that increased in the microbial community in the sediment with dense patches of the Capitella. The relationship between TOC and quinone content of the sediment as an indicator of the bacterial abundance was not clear, while a significant positive correlation was found between Capitella biomass and quinone content of the sediment. In particular, ubiquinone-10, which is present in members of the class Alphaproteobacteria, increased in the sediment with dense patches of the Capitella. We performed denaturing gradient gel electrophoresis (DGGE) analyses to identify the alphaproteobacterial species in the sediment with dense patches of the worm, using two DGGE fragments obtained from the sediment samples and one fragment from the worm body. The sequences of these DGGE fragments were closely related to the specific members of the Roseobacter clade. In the associated system with the Capitella and the bacteria in the organically enriched sediment, the decomposition of the organic matter may proceed rapidly. It is very likely that the Capitella works as a promoter of bacteria in the organically enriched sediment, and feeds the increased bacteria as one of the main foods, while the bacteria decompose the organic matter in the sediment with the assistance of the Capitella.

Microbial diversity in marine sediments from Sagami Bay and Tokyo Bay, Japan, as determined by 16S rRNA gene analysis.

16S rDNA clone libraries were analysed to investigate the microbial diversity in marine sediments from Sagami Bay (stations SA, water depth of 1159 m, and SB, 1516 m) and Tokyo Bay (station TK, 43 m). A total of 197 clones was examined by amplified rDNA restriction analysis (ARDRA) using three four-base-specific restriction enzymes (Hhal, Rsal and Haelll). In SA, 57 RFLP types were detected from 77 clones. In SB, 17 RFLP types were detected from 62 clones. In TK, 21 RFLP types were detected from 58 clones. The genotypic diversity among the three sampling sites was 0.958, 0.636 and 0.821, respectively, indicating that the microbial diversity of SA was higher than at the other two stations. At SA, the most abundant RFLP type constituted 10% of all clones. The samples from SB and TK had dominant RFLP types which constituted 60% and 38% of the total clone libraries, respectively. The community structure of SA included many single-type clones, which were found only once in the clone libraries. This structure contrasted with that of the other two stations. Thirty-seven clones were selected and sequenced according to dendrograms derived from ARDRA, to cover most of the microbial diversity in the clone libraries. No clones were identical to any of the known 165 rRNA sequences or to each other. All sequences had >84.8% similarity to rDNA sequences retrieved from the DNA databases. Sequenced clones fell into five major lineages of the domain Bacteria: the gamma, delta and epsilon Proteobacteria, Gram-positive bacteria and the division Verrucomicrobia. At SA, the Verrucomicrobia and the three subclasses of the Proteobacteria were found. Most clone sequences belonged to the gamma Proteobacteria. The high-GC Gram-positive bacteria and the gamma subclass of the Proteobacteria were common at both SB and TK. Although the depths of SB and TK were very different, the community diversity inferred from ARDRA and the taxonomic position of the dominant clones were similar. All clones belonging to the highGC Gram-positive bacteria collected from both SB and TK fell into the same cluster and are regarded as members of an unknown actinomycete group. The clone compositions were different at each sampling site, and clones of the gamma Proteobacteria and high-GC Gram-positive bacteria were dominant.