Particulate peptidoglycan in seawater determined by the silkworm larvae plasma (SLP) assay

Peptidoglycan (PG) is a biopolymer found exclusively in the cell wall of bacteria. Recent chemical analysis of particulate organic matter suggests that a major amount of the muramic acid, an amino sugar present only in PG, could not be accounted for in terms of bacterial cells (Benner and Kaiser, 2003); however, data on particulate PG is quite sparse. In the present study, conducted in 1996, the PG was examined at 5 sampling sites in the northwestern Pacific Ocean, and in natural seawater cultures. Particulate PG, which was concentrated using a 96-well filtration plate equipped with Durapore filters (pore size, 0.22 μm), was measured by the silkworm larvae plasma (SLP) assay. The PG concentration generally decreased with depth and correlated significantly with bacterial abundance throughout the entire water column. However, the ratio of particulate PG to bacterial abundance varied with depth. The average ratio was 0.61 ± 0.53 (average ± SD, n = 40) between 50 and 2000 m, which agreed with the bacterial cellular PG content from 0.63 to 1.1 fg cell⁻¹ obtained in seawater cultures. On the other hand, the ratios of PG to bacteria from the surface to 50 m (3.7 ± 2.6, n = 29) and below 2,000 m (2.1 ± 1.7, n = 7) were significantly higher than that between 50 and 2,000 m. These results may suggest that, in the surface and deep layers, a significant fraction of particulate PG was present in bacterial detritus, whereas this fraction was reduced in the middle layer.     

添加维生素B_（12）生产菌高密度培养褶皱臂尾轮虫的效果

阐述把维生素Ｂ１２生产菌添加到培养水中培养褶皱臂尾轮虫ＢｒａｃｈｉｏｎｕｓＰｌｉｃａｔｉｌｉｓ的买验。共１８株细菌分离于轮虫培养池，其中，有一株产维生素Ｂ１２的假单胞杆菌ＴＰ４对轮虫的生长繁殖有明显的促进作用。把ＴＰ４菌株培养后，加入到２Ｌ的烧杯和５００Ｌ的水槽中培养泰国Ｓ型轮虫时．在９ｄ（天）和６ｄ（天）中，轮虫密度从１２４～１３９和２４２～２８８个体／ｍｌ增殖到４，４１７～５，５４０和１，０１７～１，２５４个体／ｍｌ，分别比对照组增加了４～６及２～３倍。     

Effects of an Artificial Breakwater on the Distributions of Planktonic Microbial Communities

The summer distributions of planktonic microbial communities (heterotrophic and phtosynthetic bacteria, phtosynthetic and heterotrophic nanoflagellates, ciliate plankton, and microphytoplankton) were compared between inner and outer areas of Lake Sihwa, divided by an artificial breakwater, located on the western coast of Korea, in September 2003. The semienclosed, inner area was characterized by hyposaline surface water (<17 psu), and by low concentrations of dissolved oxygen (avg. 0.4 mg L¹) and high concentrations of inorganic nutrients (nitrogenous nutrients >36 μM, phosphate <4 μM) in the bottom layer. Higher densities of heterotrophic bacteria and nanoflagellates also occurred in the inner area than did in the outer area, while microphytoplankton (mainly diatoms) occurred abundantly in the outer area. A tiny tintinnid ciliate, Tintinnopsis nana, bloomed into more than 10⁶ cells L¹ at the surface layer of the inner area, while its abundance was much lower (10³-10⁴ cells L¹) in the outer area of the breakwater. Ciliate abundance was highly correlated with heterotrophic bacteria (r = 0.886, p < 0.001) and heterotrophic flagellates (r = 0.962, p < 0.001), indicating that rich food availability may have led to theT. nana bloom. These results suggest that the breakwater causes the eutrophic environment in artificial lakes with limited flushing of enriched water and develops into abundant bacteria, nanoflagellates, and ciliates.     

Responses of phytoplankton and heterotrophic bacteria in the northwest subarctic Pacific to in situ iron fertilization as estimated by HPLC pigment analysis and flow cytometry

To verify the hypothesis that the growth of phytoplankton in the Western Subarctic Gyre (WSG), which is located in the northwest subarctic Pacific, is suppressed by low iron (Fe) availability, an in situ Fe fertilization experiment was carried out in the summer of 2001. Changes over time in the abundance and community structure of phytoplankton were examined inside and outside an Fe patch using phytoplankton pigment markers analyzed by high-performance liquid chromatography (HPLC) and flow cytometry (FCM). In addition, the abundance of heterotrophic bacteria was also investigated by FCM. The chlorophyll a concentration was initially ca. 0.9 μg l⁻¹ in the surface mixed layer where diatoms and chlorophyll b-containing green algae (prasinophytes and chlorophytes) were predominant in the chlorophyll biomass. After the iron enrichment, the chlorophyll a concentration increased up to 9.1 μg l⁻¹ in the upper 10 m inside the Fe patch on Day 13. At the same time, the concentration of fucoxanthin (a diatom marker) increased 45-fold in the Fe patch, and diatoms accounted for a maximum 69% of the chlorophyll biomass. This result was consistent with a microscopic observation showing that the diatom Chaetoceros debilis had bloomed inside the Fe patch. However, chlorophyllide a concentrations also increased in the Fe patch with time, and reached a maximum of 2.2 μg l⁻¹ at 5 m depth on Day 13, suggesting that a marked abundance of senescent algal cells existed at the end of the experiment. The concentration of peridinin (a dinoflagellate marker) also reached a maximum 24-fold, and dinoflagellates had contributed significantly (>15%) to the chlorophyll biomass inside the Fe patch by the end of the experiment. Concentrations of 19′-hexanoyloxyfucoxanthin (a prymnesiophyte marker), 19′-butanoyloxyfucoxanthin (a pelagophyte marker), and alloxanthin (a cryptophyte marker) were only incremented a few-fold increment inside the Fe patch. On the contrary, chlorophyll b concentration reduced to almost half of the initial level in the upper 10 m water column inside the Fe patch at the end of the experiment. A decrease with time in the abundance of eukaryotic ultraphytoplankton (<ca. 5 μm in size), in which chlorophyll b-containing green algae were possibly included was also observed by FCM. Overall, our results indicate that Fe supply can dramatically alter the abundance and community structure of phytoplankton in the WSG. On the other hand, cell density of heterotrophic bacteria inside the Fe patch was maximum at only ca. 1.5-fold higher than that outside the Fe patch. This indicates that heterotrophic bacteria abundance was little respondent to the Fe enrichment.     

Mussel Periostracum from Deep-Sea Redox Communities as a Microbial Habitat: 3. Secondary Inhabitants

Abstract. Vacated microborings in periostracum of live mussels of the Florida Escarpment redox community (depth of 3266 m) become a habitat for a prolific secondary microbiota consisting of a variety of prokaryotic and a few eukaryotic organisms. Periostracum surface is also colonized by diverse microorganisms, dominated by presumed chemolithotrophic bacteria with stacks of intracellular lamellae. Unlike sheltered microflora within borings, the surface community is heavily grazed upon by numerous archaeogastropods and ciliates.     

Biological Responses to Seasonally Varying Fluxes of Organic Matter to the Ocean Floor: A Review

Deep-sea benthic ecosystems are sustained largely by organic matter settling from the euphotic zone. These fluxes usually have a more or less well-defined seasonal component, often with two peaks, one in spring/early summer, the other later in the year. Long time-series datasets suggest that inter-annual variability in the intensity, timing and composition of flux maxima is normal. The settling material may form a deposit of “phytodetritus” on the deep-seafloor. These deposits, which are most common in temperate and high latitude regions, particularly the North Atlantic, evoke a response by the benthic biota. Much of our knowledge of these responses comes from a few time-series programmes, which suggest that the nature of the response varies in different oceanographic settings. In particular, there are contrasts between seasonal processes in oligotrophic, central oceanic areas and those along eutrophic continental margins. In the former, it is mainly “small organisms” (bacteria and protozoans) that respond to pulsed inputs. Initial responses are biochemical (e.g. secretion of bacterial exoenzymes) and any biomass increases are time lagged. Increased metabolic activity of small organisms probably leads to seasonal fluctuations in sediment community oxygen consumption, reported mainly in the North Pacific. Metazoan meiofauna are generally less responsive than protozoans (foraminifera), although seasonal increases in abundance and body size have been reported. Measurable population responses by macrofauna and megafauna are less common and confined largely to continental margins. In addition, seasonally synchronised reproduction and larval settlement occur in some larger animals, again mainly in continental margin settings. Although seasonal benthic responses to pulsed food inputs are apparently widespread on the ocean floor, they are not ubiquitous. Most deep-sea species are not seasonal breeders and there are probably large areas, particularly at abyssal depths, where biological process rates are fairly uniform over time. As with other aspects of deep-sea ecology, temporal processes cannot be encapsulated by a single paradigm. Further long time-series studies are needed to understand better the nature and extent of seasonality in deep-sea benthic ecosystems. This revised version was published online in August 2006 with corrections to the Cover Date.     

Symbiotic Relations of Sediment-Agglutinating Nematodes and Bacteria in Detrital Habitats: The Enzyme-Sharing Concept

Abstract. A new concept (‘enzyme sharing’) concerning the interaction of marine nematodes and microbes in the degradation of sedimentary detritus is presented. Elements of this concept are (1) the notorious tendency of many aquatic nematodes to agglutinate detrital particles by mucus secretions, (2) new observations on the stimulation of microbial growth by nematodes in agar plates, and (3) literature data on limited endogenous proteolytic capacities of aquatic nematodes.
Observations on nematode‐microbe associations in agar plates prompted the conceptual synthesis. In agar medium without the addition of any nutrients a spectacular growth of bacteria was visible on the sinusoidal crawling trails of nematodes only 2 – 3 days after introduction of the worms (species of Adoncholaimus, Anoplostoma and Sabatieria). Juveniles of Anoplostoma that hatched in the agar cultures left their minute trails in the medium and these were rapidly occupied by bacteria. The nematodes repeatedly visited their bacterial trails, which persisted as a peculiar biotic structure for more than one year and survived the nematodes.
In sterile agar preparations containing the fluorogenic methylumbelliferyl‐β‐glucoside in the presence of the nematode Adoncholaimus, an enhanced fluorescence of the medium was visible, indicating β‐glucosidase activity. We therefore assume that oncholaimid nematodes discharge enzymes that alone, or in concert with microbial activities, contribute to the hydrolytic cleavage of refractory polysaccharides containing β‐glucosidic bonds such as agar components and cellulose. The sugars thus produced may then be taken up by the nematodes and concomitantly support the conspicuous growth of microbes. Since we did not observe any feeding of the nematodes on the associated microbes in agar plates, we question the nutritive potential of intact microbial cells for a number of nematodes abounding in detrital habitats, and call attention to the significance of ambient dissolved or adsorbed organic monomeric nutrients.
Consequently, we perceive the puzzling perpetual accretion of detrital organic particles to sediment agglutinations by nematodes as an adaptation for operating an ‘enzymatic reactor’ for the production of dissolved nutrients. We hypothesise a relationship of mutual commensalism of nematodes and heterotrophic microbes in detrital habitats and propose the term ‘enzyme sharing’ for this relationship. Both parties invest in a common enzyme pool that decomposes organic detritus for their nutrition. We present here evidence that nematodes contribute β‐glucosidase, which is involved in the cellulase system. Data from the literature suggest that microbial enzymatic processing of detrital proteins yields amino acids available to nematodes, which apparently have no efficient proteolytic enzyme system in their intestines.     

牙鲆(Paralichthys olivaceus)秦皇岛弧菌感染症及其病原细菌研究

采用病原分离与鉴定、人工感染试验等方法,在对养殖牙鲆病例进行发病情况、病理变化特征检验的基础上,进行病原学检验。结果表明,此病例属于由弧菌属(VibrioPacini1854)细菌引起的败血感染症。经对12株纯培养菌(HQ010712-1—HQ010712-12)进行形态特征、理化特性等表观分类学指征的检验,选择代表菌株(HQ010712-1株)进行16S rRNA基因的测定与系统发育学分析等,判定为弧菌属的一个新种。将HQ010712-1株送中国典型培养物保藏中心(CCTCC)进行了复核鉴定与分类定名,依据分离地定名为秦皇岛弧菌(Vibrioqinhuangdaorasp.nov.);参考菌株为HQ010712-1。     

褐藻酸降解菌侵染海带过程的超微结构观察

首次研究了褐藻酸降解菌侵入海带的途径，通过电镜观察，跟踪研究了褐藻酸降解菌染海带过程中海带表皮细胞壁的超微结构变化。结果表明，褐藻酸降解侵染海带的过程是首先引起海带表皮细胞壁藻胶层表面的破坏，然后引起藻胶层的断裂，并使藻胶层逐渐降解变突，最后褐藻酸降解菌通过海带细胞的壁藻胶层的断裂变突处进入海带细胞内。     

Geochemistry of porewater and sulfate-reducing bacteria in sediment cores from the Chianan Plain, Taiwan

High concentrations of arsenic and humic substances in groundwater from the southwestern coastal plain of Taiwan were well known for their probable relationships with black-foot disease. In order to realize the relationships between the concentrations of humic substances and arsenic in this area, 24 well water samples were analyzed. After filtered through 0.45 μm glass fiber membrane in the field, samples were kept in the dark at 4℃ and then separated into six fractions with varying range of molecular weight （〈0.5, 0.5-1, 1-5, 5-10, 10-50 and 〉50 k Da） by ultrafiltration apparatus （Molecular/Por Stirred Cell system） in the lab. Concentrations of humic substances were measured by fluorescence spectrometer （HITACHI F-2000, ex=370, em=445） and arsenic by FIAS-AA （Perkin Elmer AAnalyst 100, FIAS-400）. On average, only 6.2% of the total arsenic in water existed in the fraction of 〉0.5 k Da. and the others were complexed with humic substances （including humic acid and fulvic acid）. The results demonstrated a distinct positive correlation between the concentrations of humic substances and arsenic.