Simultaneous occurrence of denitrification and nitrate ammonification in sediments of the French Mediterranean Coast

Dissimilatory nitrate reductions in coastal marine sediment of Carteau Cove (French Mediterranean Coast) were studied between April 1993 and July 1994. Simultaneous determination of denitrification and dissimilatory nitrate reduction to ammonium was achieved by using a combination of acetylene blockage and 15N techniques. After short incubations (maximum 5 h), a part of 15N labelled nitrate added to the sediment was recovered as ammonium without incorporation in organic matter. The result indicate that a fraction of nitrate was reduced to ammonium by a dissimilatory mechanism instead of denitrifying. Denitrifying and nitrate ammonifying activities ranged from 0 to 19.8 μmol l-1 d-1 and from 2.3 to 83.2 μmol l-1 d-1, respectively. Denitrification rates were highest in early spring whereas nitrate ammonification were highest in fall. The recovery of nitrate reduced as N2O-N plus ammonium was between 40 and 100%, the highest nitrogen losses were recorded in July. Depending on the station and time of year denitrification accounted for between 0 and 43% of the total nitrate reduction whereas dissimilatory nitrate reduction to ammonium (DNRA) accounted for between 18 and 100%. The reduction rate data suggest that the pathway of nitrate reduction to ammonium may be important in coastal sediments. This revised version was published online in July 2006 with corrections to the Cover Date.     

Mixed culture of nitrifying bacteria and denitrifying bacteria for simultaneous nitrification and denitrification

A mixed culture containing nitrifying bacteria and denitrifying bacteria was investigated for aerobic simultaneous nitrification and denitrification. A mixture of NaHCO₃ and CH₃COONa was selected as the appropriate carbon source for cell growth and nitrogen removal, the concentrations of carbon and nitrogen sources were also examined. Ammonia could be oxidized aerobically to nitrite by the mixed culture, and the intermediate nitrite was then reduced to dinitrogen gas. No nitrite was detected during the process. 0.212 g of ammonia/l could be removed in 30 h and nitrate could not be utilized aerobically by the mixed culture. Nitrite could be degraded aerobically as well as anaerobically. Very little ammonia was degraded anaerobically, but the ability to degrade ammonia could be recovered even after oxygen had been supplied for 42 h.     

纳米颗粒对耐冷脱氮菌及其脱氮过程的毒害调控研究进展北大核心

生物脱氮具有经济高效、绿色环保的特点,在废水处理中具有广阔前景;目前在废水处理工艺中脱氮效果较好的微生物大部分为嗜温菌,这类菌在低温条件下的脱氮效率会受到强烈抑制,而耐冷脱氮菌可有效抵抗低温环境并进行高效脱氮,逐渐引起了研究者们的重视。其次,纳米颗粒(nanoparticles,NPs)在生物学、农学、医药等领域的应用日渐广泛,但在生产、储存和使用含NPs产品的过程中,不可避免地会向水和土壤环境释放NPs,在水环境中,NPs的大量积累会阻碍微生物的脱氮过程,也对废水处理提出了新的挑战,成为公众日益关注的环境问题和研究热点。当前,有部分研究已关注于耐冷菌的脱氮过程、NPs对耐冷菌脱氮过程的毒害作用及减毒措施。基于此,本篇文章将阐述耐冷菌的耐冷机理与脱氮过程,阐明NPs对耐冷菌脱氮过程的毒害作用与减毒调控措施,为低温环境下采用微生物处理含有NPs的氮污染废水提供理论依据。     

Effect of crude oil on denitrification and sulfate reduction in marine sediments

The denitrifying activity was measured in different types of sediment from the Mediterranean coast of France before, and after, a massive contamination (30–100 g kg^-1 sediment) of hydrocarbons. A closed system was used in order to maintain anoxic conditions and to control substrates and gaseous products concentrations. We have demonstrated that the respiratory metabolism was inhibited in all cases following an incubation time of 20 to 50 days. At this time, the addition of lactate restore the denitrifying activity. The inhibitory effect of crude oil was not related to an alteration of bacterial cells, but to changes in environmental conditions allowing denitrification. The presence of hydrocarbons in the sediments causes a decrease in the redox potential and a concomitant stimulation of the sulfate reduction.     

Similar temperature responses suggest future climate warming will not alter partitioning between denitrification and anammox in temperate marine sediments

Removal of biologically available nitrogen (N) by the microbially mediated processes denitrification and anaerobic ammonium oxidation (anammox) affects ecosystem N availability. Although few studies have examined temperature responses of denitrification and anammox, previous work suggests that denitrification could become more important than anammox in response to climate warming. To test this hypothesis, we determined whether temperature responses of denitrification and anammox differed in shelf and estuarine sediments from coastal Rhode Island over a seasonal cycle. The influence of temperature and organic C availability was further assessed in a 12‐week laboratory microcosm experiment. Temperature responses, as characterized by thermal optima (T_opt) and apparent activation energy (E_a), were determined by measuring potential rates of denitrification and anammox at 31 discrete temperatures ranging from 3 to 59 °C. With a few exceptions, T_opt and E_a of denitrification and anammox did not differ in Rhode Island sediments over the seasonal cycle. In microcosm sediments, E_a was somewhat lower for anammox compared to denitrification across all treatments. However, T_opt did not differ between processes, and neither E_a nor T_opt changed with warming or carbon addition. Thus, the two processes behaved similarly in terms of temperature responses, and these responses were not influenced by warming. This led us to reject the hypothesis that anammox is more cold‐adapted than denitrification in our study system. Overall, our study suggests that temperature responses of both processes can be accurately modeled for temperate regions in the future using a single set of parameters, which are likely not to change over the next century as a result of predicted climate warming. We further conclude that climate warming will not directly alter the partitioning of N flow through anammox and denitrification.     

深海沉积物高效脱氮菌筛选分离及其脱氮特性研究

【背景】深海海域具有高压、低温、无光等环境条件,蕴含着丰富而独特的微生物资源。【目的】从深海沉积物中定向分离、筛选脱氮效率高的好氧脱氮菌株资源,并揭示其脱氮特性,为开发水体脱氮微生物技术提供物质基础。【方法】以东太平洋、南大西洋、西南印度洋共10个站位的深海沉积物为研究材料,在28°C下使用无机氮源连续进行两轮富集培养,然后定性筛选可以脱除氨氮、亚硝态氮和硝态氮的菌株,并通过形态学和16S rRNA基因序列分析进行初步分类鉴定;对优选得到的功能菌株,分别采用以氨氮、亚硝态氮、硝态氮为唯一氮源的培养基定量研究其生长和脱氮性能。【结果】从10份大洋深海沉积物样品中共分离得到49株好氧反硝化菌,其中3株在有氧条件下反硝化效率较高,分别命名为Pseudomonassp.G111、Pseudomonassp.G112和Dietziamaris W023a,其中菌株G111和G112与模式菌株博岑假单胞菌Pseudomonas bauzanensis BZ93T的16S rRNA基因序列相似度为99.2%,菌株W023a与模式菌株海洋迪茨氏菌DietziamarisATCC35013T的16SrRNA基因序列相似度为99.9%。菌株G111、G112和W023a培养48h后,对氨氮的脱除率分别为98.0%、85.2%和97.6%;对亚硝态氮的脱除率分别为71.9%、67.5%和34.7%;对硝态氮的脱除率分别为66.0%、52.6%和56.3%。菌株G111、G112和W023a均为异养硝化-好氧反硝化菌,可通过好氧反硝化作用将亚硝态氮和硝态氮还原为含氮气体,也可通过异养硝化-好氧反硝化作用将氨氮转化为含氮气体。【结论】从深海沉积物中分离筛选得到3株高效好氧反硝化菌,所获得的菌株在水体净化、污水处理、生态系统修复等领域具有应用潜力。     

Denitrification,Nitrificastion and Nitrogen Fixation in Sublittoral Sediments of the South China Sea

Measurements of denitrification, nitrification and nitrogen fixation rates were made alongside with measuring of chemical and physical properties in sublittoral sediments of the South China Sea near the coast of Vietnam. Studied sediments were suboxic (Eh was positive as a rule), had 0.18–1.5 % of organic carbon, 0.004–0.135 % of total nitrogen and 3-12 % of total iron. The numbers of denitrifying and nitrate-reducing bacteria were as high as millions and hundreds of millions cells per gram wet weight of sediment matter, respectively. The processes of nitrification and denitrification were not spearated spacely. The nitrification was measured in both superficial layer and in a 10-cm sediment column. There were indirect evidences suggesting possibility of anaerobic ammonium oxidation. Denitrification was detectable in the sediments from two sites of sampling; maximal value was 86.2 μmoles N m⁻²h⁻¹. The denitrification potential determined at 1 mM nitrate decreased regularly from the upper to lower layers. Its values in the different sediments ranged from 134 to 532 μmoles N m⁻²h⁻¹. Nitrogen fixation (from 4.8 to 86μmoles N m⁻²h⁻¹) was close to that found in similar sediments in temperate waters in summer, and was not a significant source of nitrogen. It was comparable with diffusion of ammonium from sedimnts.     

New bacilli from shallow hydrothermal vents of Panarea Island (Italy) and their biotechnological potential

Aims: To characterize bacilli isolated from shallow hydrothermal vents of Panarea Island (Italy) and evaluate their biotechnological potential. Methods and Results: Fifteen isolates were characterized by culture and molecular methods. Eleven isolates were thermophilic, six isolates were alkalophilic and four of them were haloalkalophilic. After 16S rRNA gene sequencing, four strains, exhibiting sequence similarity below 95% with deposited strains, may represent novel species of bacilli. One strain was strictly related to Geobacillus subterraneus, but shared phenotypic characteristics for which it could be considered a new strain of this species. Four strains were affiliated with different Bacillus spp. Most isolates produced gelatinase, lipases and amylase, and some were mercury tolerant. Exopolysaccharides (EPS) production was tested adding different sugars (glucose, sucrose, trehalose, fructose, ribose, xylose and mannose, 1% w/v) as a carbon source in a minimal medium. The highest EPS yield (185 mg l^?1) was reached by strain 1A70 utilizing ribose as a carbon source. Conclusions: Novel strains of Geobacillus and indigenous ribotypes of Bacillus with biotechnological potential inhabit shallow vents of Panarea Island. Significance and Impact of the Study: New strains of thermophilic bacilli from Panarea are producers of useful biomolecules for industrial purposes as well as environmental and biotechnological applications.     

The effects of sediment desiccation on the potential for nitrification, denitrification, and methanogenesis in an Australian reservoir

Sediments from an Australian reservoir were selected for varying degrees of in situ desiccation (i.e. non-desiccated, partially desiccated and desiccated). Sediment samples were then chemically amended with appropriate electron donors and acceptors to ascertain the effect of sediment desiccation on the potential for nitrification, denitrification, methanogenesis, and the interaction of these processes. There was no detectable nitrification in these sediments yet up to 75% of added nitrate was converted to dinitrogen. Denitrification was predominantly limited by nitrate although there was evidence of carbon co-limitation. None of the nitrogen cycle processes were notably affected by sediment desiccation. There was no flush of mineral nitrogen from desiccated sediments upon rewetting. Methanogenesis did not begin in these sediments until nitrate concentrations fell below 2.25 * 10^-5 M. Methanogenesis was always carbon limited. Methanogens were affected by sediment desiccation but were capable of recovery over time upon rewetting of sediments.     

北黄海沉积物可培养产蛋白酶细菌分离鉴定

【目的】揭示北黄海沉积物中可培养产胞外蛋白酶细菌及蛋白酶多样性,增加人们对北黄海生态系统中产蛋白酶菌多样性的认识,为海洋产蛋白酶微生物的挖掘提供菌群资源。【方法】分别将5个北黄海沉积物样品梯度稀释涂布至酪蛋白明胶筛选平板,选择性分离产蛋白酶细菌;并通过分析基于16S rRNA基因序列的系统发育关系,揭示这些细菌的分类地位和遗传多样性;分别测定胞外蛋白酶活性并对酶活较高的39株菌进行基于苯甲基磺酰氟(PMSF,丝氨酸蛋白酶抑制剂)、邻菲罗啉(o-phenanthroline,O-P,金属蛋白酶抑制剂)、E-64(半胱氨酸蛋白酶抑制剂)和pepstatin A(天冬氨酸蛋白酶抑制剂)4种抑制剂的酶活抑制实验以及所有菌株对3种底物(酪蛋白、明胶、弹性蛋白)的水解能力;分析这些细菌所产胞外蛋白酶的特性及多样性。【结果】从5个北黄海沉积物样品中分离获得66株产蛋白酶细菌,这些菌株隶属于Bacteroidetes、Proteobacteria、Actinobacteria和Firmicutes 4个门的7个属,其中Pseudoalteromonas(69.9%)、Sulfitobacter(12.1%)和Salegentibacter(10.6%)是优势菌群;沉积物中可培养的产蛋白酶细菌的丰度为104 CFU/g;蛋白酶酶活抑制实验表明所有测定菌株产生的胞外蛋白酶属于丝氨酸蛋白酶和/或金属蛋白酶,仅有少数菌株所产蛋白酶具有半胱氨酸蛋白酶或天冬氨酸蛋白酶活性。【结论】北黄海沉积物中可培养产蛋白酶细菌类群较为丰富,Pseudoalteromonas、Sulfitobacter和Salegentibacter菌株是优势菌群,测定菌株所产胞外蛋白酶主要是丝氨酸蛋白酶和/或金属蛋白酶。     

Effect of bioturbation on denitrification in a marine sediment from the West Mediterranean littoral

Thein vitro effect ofNereis diversicolor on denitrification has been studied in PVC tubes filled with a coastal marine sediment defaunated by sieving. The first aim of the experiment was to determine the effect of sediment defaunation on denitrification (denitrifying population and Denitrifying Enzyme Assays). Sieving induced a loss of 70% of the initial DEA. The number of denitrifying bacteria was 10 times lower than inin situ sediment. In the top two centimetres, the DEA rose by 75% of its initial value, after 82 days. Polychaetes were only added after a return to near pre-disturbance levels to ensure that our data on the effects of their addition would not be disturbed by changes in the sediment. Introduction of Polychaetes increased the denitrifying population and DEA in the first layer (0–2 cm) of the sediment after 15 days. After 45 days, the surface of the polychaete burrows in sediment was 1.3 to 1.5 times higher than after 15 days, resulting in an increase in solute exchange between seawater and the top layer of sediment. An inhibitory effect of oxygen on denitrification was detected in the uppermost layer only.     

Abundance and diversity of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX)-metabolizing bacteria in UXO-contaminated marine sediments

Octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) is a toxic explosive known to be resistant to biodegradation. In this study, we found that sediment collected from two unexploded ordnance (UXO) disposal sites (UXO-3, UXO-5) and one nearby reference site (midref) in Hawaii contained anaerobic bacteria capable of removing HMX. Two groups of HMX-removing bacteria were found in UXO-5: group I contained aerotolerant anaerobes and microaerophiles, and group II contained facultative anaerobes. In UXO-3 and midref sediments, HMX-metabolizing bacteria were strictly anaerobic (group III and group IV). Using 16S rRNA sequencing, group I was assigned to a novel phylogenetic cluster of Clostridiales, and groups II and III were related to Paenibacillus and Tepidibacter of Firmicutes, respectively. Group IV bacteria were identified as Desulfovibrio of Deltaproteobacteria. Using [UL-(14)C]-HMX, group IV isolates were found to mineralize HMX (26.8% in 308 d) as determined by liberated (14)CO(2), but negligible mineralization was observed in groups I-III. Resting cells of isolates metabolized HMX to N(2)O and HCHO via the intermediary formation of 1-nitroso-octahydro-3,5,7-trinitro-1,3,5,7-tetrazocine together with methylenedinitramine. These experimental findings suggest that HMX biotransformation occurred either via initial denitration followed by ring cleavage or via reduction of one or more of the N-NO(2) group(s) to the corresponding N-NO bond(s) prior to ring cleavage.     

进口巴布亚新几内亚原木上截获吉丁虫的鉴定

描述江苏口岸首次从巴新原木上截获的8种吉丁虫,隶属于窄吉丁Agrilus,澳纹吉丁Anilara,星吉丁Chrysobothris,胸斑吉丁Belionota,驼翅吉丁Cyphogastra。编制族属检索表,介绍8种的鉴别特征,并配以8种吉丁虫的原色图,对检疫性害虫窄吉丁非中国种的分类状况作了简介。     

The biogeography of the yeti crabs (Kiwaidae) with notes on the phylogeny of the Chirostyloidea (Decapoda: Anomura)

The phylogeny of the superfamily Chirostyloidea (Decapoda: Anomura) has been poorly understood owing to limited taxon sampling and discordance between different genes. We present a nine-gene dataset across 15 chirostyloids, including all known yeti crabs (Kiwaidae), to improve the resolution of phylogenetic affinities within and between the different families, and to date key divergences using fossil calibrations. This study supports the monophyly of Chirostyloidea and, within this, a basal split between Eumunididae and a Kiwaidae–Chirostylidae clade. All three families originated in the Mid-Cretaceous, but extant kiwaids and most chirostylids radiated from the Eocene onwards. Within Kiwaidae, the basal split between the seep-endemic Kiwa puravida and a vent clade comprising Kiwa hirsuta and Kiwa spp. found on the East Scotia and Southwest Indian ridges is compatible with a hypothesized seep-to-vent evolutionary trajectory. A divergence date estimate of 13.4–25.9 Ma between the Pacific and non-Pacific lineages is consistent with Kiwaidae spreading into the Atlantic sector of the Southern Ocean via the newly opened Drake Passage. The recent radiation of Kiwaidae adds to the list of chemosynthetic fauna that appear to have diversified after the Palaeocene/Eocene Thermal Maximum, a period of possibly widespread anoxia/dysoxia in deep-sea basins.     

Molecular analysis of microbial communities in mobile deltaic muds of Southeastern Papua New Guinea

A PCR-based approach combined with microbiological cultivation methods was employed to determine the occurrence of sulfate-reducing bacteria (SRB) in colon biopsy samples from ulcerative colitis patients and from non-colitic controls. The detection of mucosa-associated SRB was carried out by digoxigenin-dUTP-labelled PCR amplification, in liquid Postgate medium B and in a new liquid medium, termed VM medium I. Using Postgate medium B, the growth of SRB was confirmed in 92% of the colitic specimens and in 52% of non-colitic samples. However, PCR analysis and incubation in VM medium I detected SRB in 100% of biopsy material indicating ubiquitous presence of SRB in human colon mucosa.     

人工遮光和营养添加对河流反硝化活性和反硝化细菌群落结构的影响

河流生态系统受到人类活动例如河岸带森林植被毁损和农业活动施肥等的干扰日益加剧,而这些活动使河流接收的光照增多、河流的氮磷营养盐浓度增加。微生物的反硝化作用是河流去除氮的有效途径。在汉江的一级支流金水河上游核心保护区内选取6条溪流开展野外控制实验,利用营养添加模拟河流中营养的增加,遮盖河面来模拟源头溪流的隐蔽状态,来研究河流沉积物中微生物的反硝化作用对光照和营养改变的响应,并利用高通量测序(Mi Seq)技术研究在两种处理下河流沉积物中nir S型反硝化细菌的群落结构变化。结果显示:营养元素添加促进了沉积物的反硝化活性,河面遮盖抑制了沉积物的反硝化活性。营养添加和遮盖两种处理均降低了控制实验区域内脱氯单胞菌属(优势菌属)的相对丰度,同时也降低了该区域nir S型反硝化菌群落的Chao多样性。本研究初步证实了光照增加和河流的营养增加提高了河流沉积物反硝化活性,并为提高河流的脱氮能力提供科学依据。     

The influence of nitrate concentration upon the end-products of nitrate dissimilation by bacteria in anaerobic salt marsh sediment

Abstract Experiments were carried out with slurries of saltmarsh sediment to which varying concentrations of nitrate were added. The acetylene blocking technique was used to measure denitrification by accumulation of nitrous oxide, while reduction of nitrate to nitrite and ammonium was also measured. There was good recovery of reduced nitrate and at the smallest concentration of nitrate used (250 μM) there was approximately equal reduction to either ammonium or nitrous oxide (denitrification). Nitrite was only a minor end-product of nitrate reduction. As the nitrate concentration was increased the proportion of the nitrate which was denitrified to nitrous oxide increased, to 83% at the greatest nitrate concentration used (2 mM), while reduction to ammonium correspondingly decreased. This change was attributed either to a greater competitiveness by the denitrifiers for nitrate as the ratio of electron donor to electron acceptor decreased; or to the increased production of nitrite rather than ammonium by fermentative bacteria under high nitrate, the nitrite then being reduced to nitrous oxide by denitrifying bacteria.     

Potential denitrification in submerged natural and impacted sediments of Lake Batata,an Amazonian lake

Lake Batata is one of many clear water lakes located on the floodplain of the Trombetas River in the northern Brazilian Amazon. Lake Batata is distinctly different from other lakes of this region because, for a period of 10 years, its waters received tremendous amounts of aluminum ore tailings from a bauxite mining operation. Approximately 30% of the sediments of the upper basin of the 2100 hectare lake were covered by tailings before dumping was curtailed. The goal of this research was to identify factors controlling denitrification in the natural and impacted sediments of Lake Batata. Rates of denitrification in sediments were estimated in the laboratory by the acetylene blockage method. Denitrification was measured under four conditions: without amendment; amended with glucose; amended with nitrate; and amended with glucose and nitrate. Denitrification was observed only in assays amended with nitrate suggesting that availability of nitrate is a principle factor for controlling denitrification in the sediments of Lake Batata. Effects of nitrate amendments are most pronounced when the water level is low, i.e. during the hydroperiods of draw-down and low-water.