Biomineralisation of manganese on titanium surfaces exposed to seawater

A 2-year long study was carried out to isolate and characterise various bacterial species present in the biofilm formed on titanium surfaces exposed to seawater and to assess the manganese oxidizing potential of the marine isolates. The amount of manganese present in the biofilm was also measured using atomic absorption spectrometry (AAS). The results showed that titanium was susceptible to biofouling. More than 50% of the culturable marine bacterial isolates were capable of bringing about oxidation of Mn(II). All these manganese oxidizing bacteria were heterotrophic. Autotrophic manganese oxidizing bacteria such as Leptothrix was not isolated in the present study. The AAS results confirmed that the manganese content in the biofilms increased with increasing exposure time. Hence, the study indicates that the titanium surfaces when exposed to seawater were colonised by a large number of heterotrophic bacteria, which have the ability of bringing about biomineralisation of manganese.     

Influence of urbanization on ant distribution in parks of Tokyo and Chiba City, Japan II. Analysis of species

To study the influence of urbanization on the distribution of ant species, I compared the occurrence of several ant species in urban parks of different areas and ages in Tokyo and Chiba City, Japan. Analysis of the presence or absence of 42 species revealed that the probability of occurrence in a park was significantly associated with park area for 15 species (35.7%), with park age for five (11.9%), and with region for seven (16.7%). It is likely that different species respond differently to urbanization. Species sensitive to urbanization could be good bioindicators of urban environments. Furthermore, to estimate the influence of the modification of underground environments, Messor aciculatus was selected, because this species nests in an almost vertical shaft with many chambers, to a depth of up to 4 m. The probability of occurrence of the species in a park tended to be lower in reclaimed land and banked land than in other landforms. This fact suggests that M. aciculatus could be a bioindicator of underground modification.     

Variation in sessile microflora as function of flow velocity on coupons exposed to seawater

The variability of several groups of microorganisms on AISI 1020 carbon steel coupons as a function of seawater velocity in a water circulation loop was investigated. The metal probes as well as electrodes were fixed onto ducts connected to a 35l capacity tank, in order to study both biofilm formation and some electrochemical parameters. The experiments were carried out at different seawater velocities. The technique of the most probable number was used to enumerate bacterial aerobes and anaerobes as well as sulphate-reducing bacteria and iron-reducing bacteria. Fungi were quantified by counting the number of colony forming units. At velocities of 3.6 cm/s, which correspond to a laminar flow, the numbers of aerobic and anaerobic bacteria attached to the metal surfaces reached a maximum. Such values were markedly reduced at velocities of 17.4–26.0 and 34.8 cm/s. The corrosion rate at the start of the process was 1.4 mm/year, decaying to levels of about 0.4–0.6 mm/year over the experimental period. Analysis of loss of carbon steel coupons mass after 35 days of the process indicated a mean corrosion rate of approximately 2 mm/year. This revised version was published online in November 2006 with corrections to the Cover Date.     

Seasonal and long-term changes in surface cover of aquatic plants in a shallow pond,Ojaga-ike,Chiba, Japan

Seasonal changes in surface cover and long-term floral changes in aquatic plants of a shallow eutrophic pond, Ojaga-ike, as well as the phenology of each species, were studied. Out of ten floating-leaved and submerged plants, the following five species showed a change in surface cover during 1978: Potamogeton crispus and Elodea nuttallii occupied 53.5% of the total pond surface in May, Trapa natans and Nelumbo nucifera 79.1% in July, N. nucifera and Hydrilla verticillata 62.0% in September and N. nucifera 41.2% in November. In addition to dramatic seasonal changes in surface cover, long-term floral changes occurred in this pond from 1971 onwards. While Nymphoides indica, Myriophyllum spicatum and H. verticillata decreased in number and in distribution ranges between 1971 and 1978, two newly established plants, N. nucifera and E. nuttallii, have now become dominant. The alteration in dominance may be due to water level fluctuations and interspecific competition for light. Seasonal changes in water quality during the study period are also summarized.     

Predominance of Roseobacter, Sulfitobacter, Glaciecola and Psychrobacter in seawater collected off Ushuaia, Argentina, Sub-Antarctica

Bacterial diversity in sub-Antarctic seawater, collected off Ushuaia, Argentina, was examined using a culture independent approach. The composition of the 16S rRNA gene libraries from seawater and seawater contaminated with the water soluble fraction of crude oil was statistically different (P value 0.001). In both libraries, clones representing the Alphaproteobacteria, Gammaproteobacteria, the Cytophaga-Flavobacterium-Bacteroidetes group and unculturable bacteria were dominant. Clones associated with the genera Roseobacter, Sulfitobacter, Staleya, Glaciecola, Colwellia, Marinomonas, Cytophaga and Cellulophaga were common to both the libraries. However, clones associated with Psychrobacter, Arcobacter, Formosa algae, Polaribacter, Ulvibacter and Tenacibaculum were found only in seawater contaminated with hydrocarbons (Table 1). Further, the percentage of clones of Roseobacter, Sulfitobacter and Glaceicola was high in seawater (43%, 90% and 12% respectively) compared to seawater contaminated with hydrocarbons (35%, 4% and 9% respectively). One of the clones F2C63 showed 100% similarity with Marinomonas ushuaiensis a bacterium identified by us from the same site.     

Subtype analysis and prevalence of mixed subtype infection of Blastocystis in farmed pigs from Chiba Prefecture,Japan

Blastocystis is an intestinal eukaryote found globally in humans and a wide range of animals. Blastocystis has been reported in domestic pigs, with subtype (ST) 5 being the most dominant, followed by ST1 and ST3. PCR-sequencing is commonly used for ST identification in pigs, but it often results in an underestimation of the prevalence of mixed infections. Here, we aimed to investigate the ST distribution and prevalence of mixed ST infections of Blastocystis in pigs from Chiba Prefecture in eastern Japan. A total of 82 fecal samples positive for Blastocystis were collected from two different farms, A and B. PCR was performed using ST-specific primers for ST1, ST2, ST3, and ST5. The prevalence of single ST5 infections was 37.8% (31/82), whereas that of mixed infections with ST5 and other STs was 57.3% (47/82) . A high percentage of single ST5 infections was observed in sows, piglets, and weaners from farm A (13/15, 86.7%), whereas mixed infections of ST5 and other STs (ST1 and ST3) were observed in 3- to 5-month-old grower pigs (15/18, 83.3%). Similarly, in farm B, most sows and piglets under 1 month of age showed a single ST5 infections (12/17, 70.6%), whereas weaner, grower, and finisher pigs showed mixed infections with ST5 and other STs, including ST1, ST2, and ST3 (27/28, 96.4%). In domestic pigs, diet and rearing environments change dramatically over the course of the animal's lifetime, which may have caused this difference in the prevalence of mixed ST infections among different age groups.     

Influence of urbanization on ant distribution in parks of Tokyo and Chiba City,Japan I. Analysis of ant species richness

To investigate how progressive urbanization influences the distribution of ant species in cities, I compared the ant species richness in urban parks of different areas and ages of Tokyo, the most intensively developed urban region in Japan, and its developing neighbor, Chiba City. A total of 43 ant species were found from 98 parks. Multiple regression analysis revealed that park area and age had a positive effect on the number of ant species in the parks and that the parks in Tokyo contained fewer species than did comparable parks in Chiba. Thus, the progression of urbanization reduces ant species richness in urban parks, most likely because it isolates the parks from the surrounding area. Next, in order to examine the relationships between the types of landform modification and the ant distribution in urban areas, I compared the ant species richness in urban parks of Chiba City among different landform types (upland + terraces, lowlands, cut and fill, banking, and reclamation). This study showed that the parks in the reclaimed lands and lowlands contained fewer ant species than did parks in other areas. The reason for this decline of ant fauna is discussed.     

Intestinal osmoregulatory acclimation and nitrogen metabolism in juveniles of the freshwater marble goby exposed to seawater

The objective of this study was to elucidate the role of the intestine from juveniles of the marble goby, Oxyeleotris marmorata, during seawater (SW) exposure. It has been reported elsewhere that SW-exposed juvenile O. marmorata exhibits hypoosmotic and hypoionic regulation, with the induction of branchial Na⁺/K⁺-ATPase (NKA), Na⁺:K⁺:2Cl⁻ cotransporter (NKCC), and cystic fibrosis transmembrane receptor-like chloride channels. Here, we report that SW exposure also led to significant increases in the activity and protein abundance of NKA in, and probably an increase in Na⁺ uptake through, its intestine. Additionally, there was an increase in apical NKCC immunoreactivity in the intestinal epithelium, indicating that there could be increased Cl⁻ uptake through the intestine. These results suggest that absorption of ions, and hence water, from the intestinal lumen could be an essential part of the osmoregulatory process in juvenile O. marmorata during exposure to SW. Furthermore, there were significant increases in the glutamate content, and the aminating activity and protein abundance of glutamate dehydrogenase (GDH) in the intestine of fish exposed to SW. Since the intestinal glutamine synthetase activity and protein abundance decreased significantly, and the intestinal glutamine content remained unchanged, in the SW-exposed fish, excess glutamate formed via increased GDH activity in the intestine could be channeled to other organs to facilitate the increased synthesis of amino acids. Taken together, our results indicate for the first time that, besides absorbing ions and water during SW exposure, the intestine of juvenile O. marmorata also participated in altered nitrogen metabolism in response to salinity changes.     

Teratogenic, biochemical, and histological studies with mice prenatally exposed to 2.45-GHz microwave radiation

Pregnant CD-1 mice were exposed to 2.45-GHz continuous wave microwave radiation at an incident power density of 30 mW/cm2. The local specific absorption rate near the uterine area (deep colonic location), as determined from time-temperature profiles measured with a Vitek thermistor probe, was 40.2 mW/g. Groups of mice were exposed 8 hr per day through Days 1-6 or 6-15 of pregnancy. Other groups of animals were exposed to an elevated ambient temperature of 31 degrees C which increased the colonic temperature 2.3 degrees C, the same as that produced by the microwaves. Sham-irradiated groups of animals were treated exactly the same as the microwave-exposed animals. For the two conditions, temperature exposed and sham exposed, two groups of animals were used. One group was handled in the same manner as the microwave-irradiated group and the other group was not handled so as to evaluate the effects of stressing the animals by handling. Eleven groups of animals were used in the complete study: five groups for gestational Days 1-6, five groups for gestational Days 6-15, and one group of cage control animals. On Day 18 of gestation the dams of all experimental groups were sacrificed and their reproductive status was determined. The fetuses were examined for visceral and skeletal alterations. Brain cholinesterase activity and histology were evaluated in the groups exposed on Days 6-15. The results show that microwave radiation increases embryo lethality at the early stages of gestation (exposure Days 1-6). Fetal toxicity and teratogenicity were not significantly increased by exposure to microwaves on either Days 1-6 or 6-15 of gestation. Cholinesterase activity and histology of the brain of 18-day-old fetuses were not adversely affected.     

Genetic identification of fish eggs collected in Sendai Bay and off Johban,Japan

Mitochondrial DNA typing of 52 fish eggs collected in Sendai Bay and off Johban revealed ten haplotypes of the 225–251 bp region of 16S ribosomal DNA. Comparing with known sequences of fishes, we identified them as Trachurus japonicus (Carangidae), Pagrus major, Evynnis japonica (Sparidae), Paralichthys olivaceus, Pseudorhombus sp. (Paralichthidae), Pennahia argentata (Sciaenidae) and Platycephalus sp. (Platycephalidae). The occurrence of Trachurus japonicus and sparid eggs in the study area indicates significant extensions to the ranges of their spawning sites northward. Our method is useful for identifying fish eggs that are difficult to diagnose by morphology, and to uncover cryptic fish egg assemblages in the field.     

Historical view of DNA replication studies, with special reference to Japan

Almost forty years after the key contributions to the field by Okazaki and coworkers that gave rise to the concept of leading and the lagging strand, we are still at the state of uncertainty about the proteins that replicate each strand. Perhaps, one main conclusion that should be drawn from the data currently available is that the protein architecture at the fork is more plastic than originally thought.     

Persistence of Vibrio vulnificus in tissues of Gulf Coast oysters, Crassostrea virginica, exposed to seawater disinfected with UV light.

Vibrio vulnificus is an estuarine bacterium which can cause opportunistic infections in humans consuming raw Gulf Coast oysters, Crassostrea virginica. Although V. vulnificus is known as a ubiquitous organism in the Gulf of Mexico, its ecological relationship with C. virginica has not been adequately defined. The objective of the present study was to test the hypothesis that V. vulnificus is a persistent microbial flora of oysters and unamenable to traditional methods of controlled purification, such as UV light depuration. Experimental depuration systems consisted of aquaria containing temperature-controlled seawater treated with UV light and 0.2-microns-pore-size filtration. V. vulnificus was enumerated in seawater, oyster shell biofilms, homogenates of whole oyster meats, and tissues including the hemolymph, digestive region, gills, mantle, and adductor muscle. Results showed that depuration systems conducted at temperatures greater than 23 degrees C caused V. vulnificus counts to increase in oysters, especially in the hemolymph, adductor muscle, and mantle. Throughout the process, depuration water contained high concentrations of V. vulnificus, indicating that the disinfection properties of UV radiation and 0.2-microns-pore-size filtration were less than the rate at which V. vulnificus was released into seawater. Approximately 10(5) to 10(6) V. vulnificus organisms were released from each oyster per hour, with 0.05 to 35% originating from shell surfaces. These surfaces contained greater than 10(3) V. vulnificus organisms per cm2. In contrast, when depuration seawater was maintained at 15 degrees C, V. vulnificus was not detected in seawater and multiplication in oyster tissues was inhibited.     

Persistence of Vibrio vulnificus in tissues of Gulf Coast oysters, Crassostrea virginica, exposed to seawater disinfected with UV light.

Vibrio vulnificus is an estuarine bacterium which can cause opportunistic infections in humans consuming raw Gulf Coast oysters, Crassostrea virginica. Although V. vulnificus is known as a ubiquitous organism in the Gulf of Mexico, its ecological relationship with C. virginica has not been adequately defined. The objective of the present study was to test the hypothesis that V. vulnificus is a persistent microbial flora of oysters and unamenable to traditional methods of controlled purification, such as UV light depuration. Experimental depuration systems consisted of aquaria containing temperature-controlled seawater treated with UV light and 0.2-microns-pore-size filtration. V. vulnificus was enumerated in seawater, oyster shell biofilms, homogenates of whole oyster meats, and tissues including the hemolymph, digestive region, gills, mantle, and adductor muscle. Results showed that depuration systems conducted at temperatures greater than 23 degrees C caused V. vulnificus counts to increase in oysters, especially in the hemolymph, adductor muscle, and mantle. Throughout the process, depuration water contained high concentrations of V. vulnificus, indicating that the disinfection properties of UV radiation and 0.2-microns-pore-size filtration were less than the rate at which V. vulnificus was released into seawater. Approximately 10(5) to 10(6) V. vulnificus organisms were released from each oyster per hour, with 0.05 to 35% originating from shell surfaces. These surfaces contained greater than 10(3) V. vulnificus organisms per cm2. In contrast, when depuration seawater was maintained at 15 degrees C, V. vulnificus was not detected in seawater and multiplication in oyster tissues was inhibited.     

Deposition of fibronectin on material surfaces exposed to plasma: quantitative and biological studies

Experiments were carried out to characterize plasma fibronectin deposition onto material surfaces exposed to plasma solutions. Under nonclotting conditions, the amount of fibronectin adsorption on the surfaces, determined by an indirect radioactive antibody assay, was maximal at low plasma concentrations (0.1%). At higher concentrations of plasma, other plasma proteins appeared to compete with and inhibit adsorption of fibronectin. Biological activity (fibronectin-promoted cell spreading) was also greatest at low plasma concentrations and decreased as the plasma concentration was raised. When surfaces were exposed to plasma under clotting conditions (i.e., addition of Ca2+ and thrombin), fibronectin deposition on the surfaces and biological activity remained constant or increased as the plasma concentration was raised. Based on indirect immunofluorescent antibody assays, the fibronectin deposited from clotting plasma appeared to be in a punctate distribution over the entire material surface and occasionally was associated with discrete fibrillar structures. The increased deposition of fibronectin from clotting plasma compared to nonclotting plasma (approximately a 10-fold difference with 10% plasma) was partially a result of covalent crosslinking of fibronectin to fibrin based upon studies with putrescine added to inhibit crosslinking during clotting. On the other hand, the increase in biological activity that occurred if the surfaces were exposed to clotting plasma was completely inhibited by putrescine, indicating that fibronectin had to be crosslinked to fibrin to have biological activity under these conditions. Finally, fibronectin deposition also occurred on surfaces exposed to whole blood and was markedly enhanced when clotting occurred.     

Generalization of learned predator recognition: an experimental test and framework for future studies

While some prey species possess an innate recognition of their predators, others require learning to recognize their predators. The specific characteristics of the predators that prey learn and whether prey can generalize this learning to similar predatory threats have been virtually ignored. Here, we investigated whether fathead minnows that learned to chemically recognize a specific predator species as a threat has the ability to generalize their recognition to closely related predators. We found that minnows trained to recognize the odour of a lake trout as a threat (the reference predator) generalized their responses to brook trout (same genus as lake trout) and rainbow trout (same family), but did not generalize to a distantly related predatory pike or non-predatory suckers. We also found that the intensity of antipredator responses to the other species was correlated with the phylogenetic distance to the reference predator; minnows responded with a higher intensity response to brook trout than rainbow trout. This is the first study showing that prey have the ability to exhibit generalization of predator odour recognition. We discuss these results and provide a theoretical framework for future studies of generalization of predator recognition.     

日本北海道地区细石叶技术研究概述

细石叶技术是晚更新世晚期东北亚和西北美新的变革性剥坯技术和适应手段。北海道地区拥有丰富、多样的细石器遗存,相关材料最早可以追溯到距今约2.5万年前,持续了约1.2万年。以北海道具有明显技术复杂性和多样性特点的细石器为材料,学者们基于技术式"阅读"和拼合研究划分出一系列细石核工艺类型和剥坯技法;岛内以楔形细石核为代表的细石器工业传统在不断延续的同时,也表现出明显的阶段性变化,并在一定程度上显示出与气候环境变化较强的相关性。本文对北海道地区晚更新世末次盛冰期以来的气候、环境背景以及细石器的发现与研究历程进行介绍。在此基础上,厘清北海道细石核工艺类型的划分,并对细石器群的阶段性演变进行系统梳理,以期对我国相关研究有所裨益。