New Discoveries in Study on Hydrocarbons From Thermal Degradation of Heterotrophically Yellowing Algae

Green autotrophic alga Chlorella protothecoides contains a very small quantity of hydrocarbons. Heterotrophic culture of this alga results in the cells yellowing, chlorophyll disappearing, protein decreasing and lipid increasing remarkably. The quantities of hydrocarbons from them directly and from the thermal degradation of the cells at or below 200℃ are very low. These hydrocarbons are characterized by predominance of high molecular weight normal alkanes with maximum at C23-C25. When these heterotrophi-cally yellowing cells are thermally degraded at 300℃ , the aliphatic hydrocarbons increase greatly, 32 times that of the green autotrophic ones at the same temperature. Meanwhile, the low molecular weight normal alkanes with C17 as the peak become predominant instead of the original ones of high molecular weight. The actual potential of microplanktonic algae in producing hydrocarbons should be much greater than what people have recognized before.

Influences of Bacterial Degradation on Gases Generated by Pyrolysis of Heterotrophically Yellowing Algae

The large amount of microplanktonic algae in nature are not all thermally degraded directly into organic matter from their original green autotrophic state. When the algae are old or dead and deposited at the bottom of lake or ocean, some changes may take place in the cells, including the disappearance of the chlorophyll, the

Salinity Acclimation Induces Reduced Energy Metabolism,Osmotic Pressure Regulation and Transcriptional Reprogramming in Hypotrichida Ciliate Gastrostyla setifera

Coastal and estuarine protists are frequently exposed to salinity undulation.While the tolerance and stress responses of microalgae to salinity have been extensively studied,there have been scarce studies on the physiological response of heterotrophic protists to salinity stressing.In this study,we investigated the physiological response of the heterotrophic ciliate Gastrostyla setifera to a salinity of 3,via a transcriptomic approach.The first transcriptome of genus Gastrostyla was obtained utilizing a group of manually isolated ciliate individuals(cells)and RNA-seq technique.The completeness of the transcriptome was verified.Differentially expressed gene(DEG)analysis was performed among the transcriptomes of G.setifera acclimated in saline water(salinity 3)and those cultured in fresh water.The results demonstrated a significant alternation in gene transcription,in which the ciliate exhibits a transcripttomic acclimation in responding salinity stressing.The up-regulated DEGs were enriched in the pathways of cytoskeleton proteins,membrane trafficking,protein kinases and protein phosphatases.These may represent enhanced functions of ion transport,stress response and cell protections.Pathways involved in energy metabolism and biosynthesis were markedly down-regulated,reflecting decreased cell activity.Particularly,we detected significantly down-regulated genes involved in several pathways of amino acid catabolism,which may lead to accumulation of amino acids in the ciliate cell.Amino acid could act as compatible solutes in the cytoplasm to maintain the osmotic balance in saline water.Overall,this work is an initial exploration to the molecular basis of the heterotrophic protist responding to salinity stressing.The result sheds light on the mechanisms of enhancement of cell protection,reduction of cell activity,and osmotic pressure regulation in ciliates acclimated to salinity.

Long-term thinning decreases the contribution of heterotrophic respiration to soil respiration in subalpine plantations

Interest in the dynamics of soil respiration(R_(S))in subalpine forest ecosystems is increasing due to their high soil carbon density and potential sensitivity to environmental changes.However,as a principal silvicultural practice,the long-term impacts of thinning on R_(S) and its heterotrophic and autotrophic respiration components(R_(h) and Ra,respectively)in subalpine plantations are poorly understood,espe-cially in winter.A 3-year field observation was carried out with consideration of winter CO_(2) efflux in middle-aged sub-alpine spruce plantations in northwestern China.A trench-ing method was used to explore the long-term impacts of thinning on Rs,Rn and R_(a).Seventeen years after thinning,mean annual Rs,Rn and R_(a) increased,while the contribu-tion of R_(h) to R_(s) decreased with thinning intensity.Thinning significantly decreased winter R,because of the reduction in R_(n) but had no significant effect on Ra.The temperature sensitivity(Q_(10))of R_(h) and R_(a) also increased with thinning intensity,with lower Q_(10) values for R_(h)(2.1-2.6)than for Ra(2.4-2.8).The results revealed the explanatory variables and pathways related to R_(n) and R_(a) dynamics.Thinning increased soil moisture and nitrate nitrogen(NO_(3)^(-)-N),and the enhanced nitrogen and water availability promoted R_(h) and R_(a) by improving fine root biomass and microbial activity.Our results highlight the positive roles of NO_(3)^(-)-N in stimulating R_(s) components following long-term thinning.Therefore,applications of nitrogen fertilizer are not recommended while thinning subalpine spruce plantations from the perspective of reducing soil CO_(2) emissions.The increased Q_(10) values of R_(s) components indicate that a large increase in soil CO_(2) emissions would be expected following thinning because of more pronounced climate warming in alpineregions.

Improving dryland maize productivity and water efficiency with heterotrophic ammonia-oxidizing bacteria via nitrification and cytokinin activity

A two-year field experiment conducted under dryland conditions in semi-humid and drought-prone regions of China aimed to assess the effect of ammonia-oxidizing bacterial on maize water use efficiency and yield.A heterotrophic ammonia-oxidizing bacteria(HAOB)strain S2_8_1 was used.Six treatments were applied:(1)no irrigation+HAOB strain(DI),(2)no irrigation+blank culture medium(DM),(3)no irrigation control(DCK),(4)irrigation+HAOB(WI),(5)irrigation+blank culture medium(WM),and(6)irrigation control(WCK).Results revealed that HAOB treatment increased maize growth,yield,and water use efficiency over controls,regardless of whether the year was wet or dry.This improvement was attributed to the accelerated nitrification in the rhizosphere soil due to HAOB inoculation,which subsequently led to increased levels of leaf cytokinins.Overall,these findings suggest that HAOB inoculation holds promise as a strategy to boost water use efficiency and maize productivity in dryland agriculture.

缢蛏滩涂养殖环境的细菌群落组成及分析

In this study,the bacteria from the mud in tidal-flat Sinonovacula constricta aquaculture area were isolated each month from March to December,2002,and the temporal and spatial distribution of heterotrophic bacteria,ammonifying bacteria,denitrifying bacteria,and sulphate reducing bacteria were analyzed.The results showed that all the 515 isolated bacteria mainly belonged to 1 family and 13 genera.The bacterial flora in different layers of the mud was almost consistent,while the composition was different.The predominant genera were Clostridium,Bacillus,Corynebacterium,Photobacterium,and some Enterobacteriaceae.The number of heterotrophic bacteria in the surface layer and the bottom fluctuated in 7.6×103 cfu·g-1～2.0×105 and 1.6×103～1.0×105 cfu·g-1,ammonifying bacteria fluctuated in 1.5×106～9.0×107 and 9.0×105～1.0×107 cfu·g-1,denitrifying bacteria fluctuated in 9.0×103～4.0×106 and 5.0×102～1.9×106 cfu·g-1,and sulphate reducing bacteria fluctuated in 5.0×104～5.0×106 and 1.9×104～2.0×106 cfu·g-1,respectively.The detection rates of ammonifying bacteria,denitrifying bacteria and sulphate reducing bacteria in the mud were all 100%,and these bacteria increased significantly in the second half of the year,indicating that the environment of the Sinonovacula constricta aquaculture area was deteriorated due to the accumulation of NH3,nitrite and H2S,and it is important to regulate the breed capacity and redistribute the breeding environment.

Interannual Variation in Terrestrial Ecosystem Carbon Fluxes in China from 1981 to 1998

A dynamic biogeochemical model was used to estimate the responses of China's terrestrial net primary productivity (NPP), soil heterotrophic respiration (HR) and net ecosystem productivity (NEP) to changes in climate and atmospheric CO2 from 1981 to 1998. Results show that China's total NPP varied between 2.89 and 3.37 Gt C/a and had an increasing trend by 0.32% per year, HR varied between 2.89 and 3.21 Gt C/a and grew by 0.40% per year, Annual NEP varied between -0.32 and 0.25 Gt C but had no statistically significant interannual trend. The positive mean NEP indicates that China's terrestrial ecosystems were taking up carbon with a total carbon sequestration of 1.22 Gt C during the analysis period. The terrestrial NEP in China related to climate and atmospheric CO2 increases accounted for about 10% of the world's total and was similar to the level of the United States in the same period. The mean annual NEP for the analysis period was near to zero for most of the regions in China, but significantly positive NEP occurred in Northeast China Plain, the southeastern Xizang (Tibet) and Huang-Huai-Hai Plain, and negative NEP occurred in the Da Hinggan Mountains, Xiao Hinggan Mountains, Loess Plateau and Yunnan-Guizhou Plateau. China's climate at the time was warm and dry relative to other periods, so the estimated NEP is probably lower than the average level. China's terrestrial NEP may increase if climate becomes wetter but is likely to continue to decrease if the present warming and drying trend sustains.

Study on the Quantity Dynamic Changes of Heterobacteria and Vibrios in Larvae Industrialized Culture System

[ Objective ] The dynamic change of heterobacteria and vibrios in larvae industrialized culture system was studied to provide scientific reference for healthy cultivation of shrimp. [ Method ] The heterobacteria, vibrios and pathogenic vibrio parahaemolyticus were monitored in larvae industrialized culture system. [ Result] The heterobacteria, vibrios and pathogenic vibrio parahaemolyticus were the most in fertilized eggs of shrimp but the least in nauplius, then their number would increase with growth. During whole rearing period, both boterobacteria in larvae, vibrios in water would increase by one order of magnitude, while both vibrios in larvae and heterobacteria in water would increase by two orders of magnitude. There were many heterobacteria and vibrios but few vibrio parahaemolyticus in living bait. The correlation coefficients between larvae and heterobacteria and vibrios in water were 0. 704 and 0. 840 in culture system respectively, while the correlation among heterobacteria, vibrios in living bait and larvae, water were weak or negative. [ Conclusion ] There was a dynamic relation between water and larvae in rearing period, and restrictly control of culture condition would restrain the occurrence of disease caused by vibrio parahaemolyticus, besides that bacteria number in bait was not obviously correlated with bacteria nubmer in culture system.

Bioleaching of heavy metals from contaminated alkaline sediment by auto- and heterotrophic bacteria in stirred tank reactor

Bioleaching Xiangjiang River alkaline sediment contaminated by multiple heavy metals was investigated. Multiple metals in alkaline sediment possess significant toxicity to aquatic organisms or humans and will greatly inhibit bioleaching. The bioleaching method using autotrophic bacteria mixed with heterotrophic bacteria can solve this problem successfully. The experiment results showed that bioleaching efficiencies of Zn, Mn, Cu, and Cd were 95.2 %, 94.2 %, 90.1 %, and 84.4 %, respectively. Moreover, the changes of heavy metal concentrations in different fractions in contaminated sediment before and after bioleaching were analyzed by selective sequential extraction, and it was discovered that the main fractions of Zn, Mn, Cu and Cd after bioleaching are Fe-Mn oxide, organic associated form and a residual form. Its biotoxicity decreased greatly. The bioleaching heavy metals from sediment using autotrophic bacteria combined with heterotrophic bacteria can effectively improve the bioleaching efficiency and reduce toxicity.

Soil heterotrophic respiration in Casuarina equisetifolia plantation at different stand ages

The soil respiration rates （Rh） in 6-year-old （young）, 17-year-old （middle-age）, 31-year-old （mature） Casuarina equisetifolia coastal plantations were measured using an LICOR-8100 automated soil CO2 flux system from May 2006 to April 2007. Results show that Rh displayed an obvious seasonal pattern across the observed years. The maximum values of Rh occurred at June and July and the minimum at December and January. Soil temperature and soil moisture as well as their interaction had significant effects on the monthly dynamics of Rh. The analysis by one-way ANOVA showed that Rh had a significantly exponential relation （p〈0.05） to soil temperature at soil depth of 5 cm, and had a linear relation （p〈0.05） to soil water content of the upper 20 cm. The result estimated by the two-factor model shows that soil temperature at soil depth of 5 cm and soil moisture at soil depth of 20 cm could explain 68.9%-91.9% of seasonal variations in Rh. The or- der of Rh rates between different stand ages was middle-age plantation〉mature plantation〉young-age plantation. With the increase of growth age of plantation, the Q10 of Rh increased. The contribution of Rh to total soil surface CO2 flux was 71.89%, 71.02% and 73.53% for the young, middle-age and mature plantation, respectively. It was estimated that the annual CO2 fluxes from Rh were 29.07, 38.964 and 30.530 t.ha^-1.a^-1 for the young, middle-age and mature plantation, respectively.

Discussion on Problem of Fermentation Contamination of Heterotrophic Chlorella

The contamination reasons were analyzed form the viewpoints of strain preparation, air system, medium and equipment sterilization and pipeline configura- tion. Moreover, the contamination-preventing measures were defined. We also sum- marized the characteristics of bacteria that contaminated the fermentation, as well as checking methods and remedial measures.

Heterotrophic ammonium removal characteristics of an aerobic heterotrophic nitrifying-denitrifying bacterium, Providencia rettgeri YL

Bacterium Providencia rettgeri YL was found to exhibit an unusual ability to heterotrophically nitrify and aerobically denitrify various concentrations of ammonium （NH4^＋-N）. In order to further understand its removal ability, several experiments were conducted to identify the growth and ammonium removal response at different carbon to nitrogen （C/N） mass ratios, shaking speeds, temperatures, ammonium concentrations and to qualitatively verify the production of nitrogen gas using gas chromatography techniques. Results showed that under optimum conditions （C/N 10, 30℃, 120 r/min）, YL can significantly remove low and high concentrations of ammonium within 12 to 48 h of growth, respectively. The nitrification products hydroxylamine （NHzOH）, nitrite （NO2^-） and nitrate （NO3^-） as well as the denitrification product, nitrogen gas （N2）, were detected under completely aerobic conditions.

Short-term effects of nitrogen deposition on soil respiration components in two alpine coniferous forests, southeastern Tibetan Plateau

Nitrogen (N) deposition to alpine forest ecosystems is increasing gradually, yet previous studies have seldom reported the effects of N inputs on soil CO2 flux in these ecosystems. Evaluating the effects of soil respiration on N addition is of great significance for understanding soil carbon (C) budgets along N gradients in forest ecosystems. In this study, four levels of N (0, 50, 100, 150 kg N ha^-1 a^-1) were added to soil in a Picea baifouriana and an Abies georgei natural forest on the Tibetan Plateau to investigate the effect of the N inputs on soil respiration. N addition stimulated total soil respiration (Rt) and its components including heterotrophic respiration (Rh) and autotrophic respiration (Ra);however, the promoted effects declined with an increase in N application in two coniferous forests. Soil respiration rate was a little greater in the spruce forest (1.05 μmol CO2 m^-2 s^-1) than that in the fir forest (0.97 μmol CO2 m^-2 s^-1). A repeated measures ANOVA indicated that N fertilization had significant effects on Rt and its components in the spruce forest and Rt in the fir forest, but had no obvious effect on Rh or Ra in the fir forest. Rt and its components had significant exponential relationships with soil temperature in both forests. N addition also increased temperature sensitivity (Q10) of Rt and its components in the two coniferous forests, but the promotion declined as N in put increased. Important, soil moisture had great effects on Rt and its components in the spruce forest (P<0.05), but no obvious impacts were observed in the fir forest (P>0.05). Following N fertilization, Ra was significantly and positively related to fine root biomass, while Rh was related to soil enzymatic activities in both forests. The mechanisms underlying the effect of simulated N deposition on soil respiration and its components in this study may help in forecasting C cycling in alpine forests under future levels of reactive N deposition.

Enhanced biological nutrient removal by the alliance of a heterotrophic nitrifying strain with a nitrogen removing ecosystem

Nitrogen removal from synthetic wastewater was investigated in an airlift bioreactor （ALB）, augmented with a novel heterotrophic nitrifier Pseudonocardia ammonioxydans H9^T under organic carbon to nitrogen ratios （Corg/N） ranging from 0 to 12. Effect of the inoculated strain was also determined on the settling properties and the removal of chemical oxygen demand （COD）. Two laboratory scale reactors were set up to achieve a stable nitrifying state under the same physicochemical conditions of hydraulic retention time （HRT）, temperature, pH and dissolved oxygen （DO）, and operated under the sequencing batch mode. The level of DO was kept at 0.5- 1.5 mg/L by periodic stirring and aeration. Each specific Corg/N ratio was continued for duration of 3 weeks. One of the reactors （BR2） was inoculated with P ammonioxydans H9^T periodically at the start of each Corg/N ratio. Sludge volumetric index （SVI） improved with the increasing Corg/N ratio, but no significant difference was detected between the two reactors. BR2 showed higher levels of nitrogen removal with the increasing heterotrophic conditions, and the ammonia removal reached to the level of 82%-88%, up to10% higher than that in the control reactor （BR1） at Corg/N ratios higher than 6; however, the ammonia removal level in experimental reactor was up to 8% lower than that in control reactor at Corg/N ratios lower than 2. The COD removal efficiency progressively increased with the increasing Corg/N ratios in both of the reactors. The COD removal percentage up to peak values of 88%-94% in BR2, up to 11% higher than that in BR1 at Corg/N ratio higher than 4. The peak values of ammonia and COD removal almost coincided with the highest number （18%-27% to total bacterial number） of the exogenous bacterium in the BR2, detected as colony forming units （CFU）. Furthermore, the removal of ammonia and COD in BR2 was closely related to the number of the inoculated strain with a coefficient index （R2） up to 0.82 and 0.85 for ammonia and COD, respectively. These results suggest that it was more efficient for both the ammonia and carbon nutrient removals in a reactor inoculated with a heterotrophic nitrifier at high Corg/N ratio, inferring that the heterotrophic nitrifers would be practically more available in the treatment of wastewater with high level of ammonia and COD.

Effect of carbon source on growth and lipid accumulation in Chlorella sorokiniana GXNN01

Heterotrophic culture of microalgae to develop methods of increasing biomass productivity and storage lipids has brought new insight to commercial biodiesel production. To understand the relationship between heterotrophy and lipid production, the effects of carbon sources on the growth and lipid accumulation of Chlorella sorokiniana GXNN01 was studied. The alga exhibited an increased growth rate in response to the addition of carbon sources, which reached the stationary phase after 48 h at 30℃. In addition, glucose and NaAc had a significant effect on the lipid accumulation during the early-stationary phase. Specifically, the lipid content was 0.237±0.026 g ·g^-1 cell dry weight and 0.272±0.041 g Ll when glucose was used as the carbon source, whereas the lipid content reached 0.287±0.018 g ·g^-1 cell dry weight and 0.2884-0.008 g Lz when NaAc was used as the carbon source. The neutral lipid content was found to first decrease and then increase over time during the growth phase. A glucose concentration of 20 mmol L^-1 gave the maximal lipid yield and the optimum harvest time was the early-stationary phase.

Effect of ultraviolet mutagenesis on heterotrophic strain mutation and bioleaching of low grade copper ore

The effect of ultraviolet mutagenesis on a heterotrophic strain(Providencia JAT-1) mutation was studied and bioleaching of low grade copper ore with mutant bacteria was investigated. The results show that the activity of bacteria was improved after ultraviolet mutagenesis; the best irradiation time was 120 s. Compared to the original bacteria, the cells density of mutant bacteria at stationary phase increased by 26% and ammonia produced by mutant bacteria increased by 12%. Higher activity of bacteria leads to a higher copper extraction rate. The bioleaching performance of Providencia JAT-1 was improved after UV mutagenesis. The copper extraction rate with mutant bacteria increased by 10.6% compared to the original bacteria. The ore surface was corroded and the fine particles were absent after bioleaching. Free copper oxide and copper silicates could be leached out easily by using JAT-1; a small part of the copper sulfide can also be leached out. Bioleaching using JAT-1 is more effective than ammonia leaching and copper extraction rate with mutant bacteria was 21.1% higher than that by ammonia leaching under the same condition.

Removal of Nitrogen, Phosphorus, and Organic Pollutants From Water Using Seeding Type Immobilized Microorganisms

Objective To study the possibility of removing nitrogen, phosphorus, and organic pollutants using seeding type immobilized microorganisms. Methods Lakes P and M in Wuhan were chosen as the objects to study the removal of nitrogen, phosphorus, and organic pollutants with the seeding type immobilized microorganisms. Correlations between the quantity of heterotrophic bacteria and the total nitrogen （TN）, total phosphorus （TP）, and total organic carbon （TOC） in the two lakes were studied. The dominant bacteria were detected, inoculated to the sludge and acclimated by increasing nitrogen, phosphorus and decreasing carbon source in an intermittent, time-controlled and fixed-quantity way. The bacteria were then used to prepare the seeding type immobilized microorganisms, selecting diatomite as the adsorbent cartier. The ability and influence factors of removing nitrogen, phosphorus, and organic pollutant from water samples by the seeding type immobilized microorganisms were studied. Results The coefficients of the heterotrophic bacterial quantity correlated with TOC, TP, and TN were 0.9143, 0.8229, 0.7954 in Lake P and 0.9168, 0.7187, 0.6022 in Lake M. Ten strains of dominant heterotrophic bacteria belonging to Pseudomonas, Coccus, Aeromonas, Bacillus, and Enterobateriaceae, separately, were isolated. The appropriate conditions for the seeding type immobilized microorgansims in purifying the water sample were exposure time=24 h, pH=7.0-8.0, and quantity of the immobilized microorganisms=0.75-1g/50 mL. The removal rates of TOC, TP, and TN under the above conditions were 80.2%, 81.6%, and 86.8%, respectively. Conclusion The amount of heterotrophic bacteria in the two lakes was correlated with TOC, TP, and TN. These bacteria could be acclimatized and prepared for the immobilized microorganisms which could effectively remove nitrogen, phosphorus, and mixed organic pollutants in the water sample.

Dynamics of picoplankton in the Nansha Islands area of the South China Sea

Dynamics of major picoplankton groups, Synechococcus (Syn), Prochlorococcus (Pro), picoeukaryotes (Euk) and heterotrophic bacteria (Bact) was investigated by flow cytometry for the first time in the Nansha Islands area in the South China Sea. Averaged over the whole investigation area, depth-weighted integrated cell abundance (DWA) of Syn, Pro, Euk and Bact was 1.6 (0.4-5.7)×103, 5.4 (0.1-7.3)×104, 0.7 (0.2-2.2)×103, and 2.3 (1.4-3.2)×105 cells/mL respectively. Picoautotrophic cell abundance was low in the northwest part of the Nansha Islands where surface water temperature was low and the upper mixed layer was shallow. Concurrently, a surface maximum vertical distribution pattern was observed in this area. While in the southeast and east zones where temperatures were relatively higher and nitraclines were deeper, picoplankton is abundant and a subsurface maximum around 50-75 m is observed. Coupling of horizontal and vertical distribution patterns of picoplankton abundance and hydrological status was found, suggesting a strong influence of currents and water column structure on picoplankton distribution in the investigation area. Contrary to that in the shelf water in the East China Sea, the relationship between Pro and Bact in the Nansha Islands area in the South China Sea was not significantly negative but weakly positive. Moreover, a similar distribution pattern of Syn and Pro was observed. Possible reasons for these differences in the two marine regimes were discussed.

Purification and Characterization of a Low-temperature Hydroxylamine Oxidase from Heterotrophic Nitrifier Acinetobacter sp.Y16

Objective To purify a low-temperature hydroxylamine oxidase （HAO） from a heterotrophic nitrifying bacterium Acinetobacter sp. Y26 and investigate the enzyme property. Methods A HAO was purified by an anion-exchange and gel-filtration chromatography from strain Y16. The purity and molecular mass were determined by RP-HPLC and SDS-PAGE. The HAO activity was detected by monitoring the reduction of potassium ferricyanide using hydroxylamine as substrate and ferricyanide as electron acceptor. The partial amino acid sequence was determined by mass spectrometry. Results The low-temperature HAO with a molecular mass of 61 kDa was purified from strain Y26 by an anion-exchange and gel-filtration chromatography. The enzyme exhibited an ability to oxidize hydroxylamine in wide temperature range （4-40 ℃） in vitro using hydroxylamine as substrate and ferricyanide as electron acceptor. It was stable in the temperature range of 4 to 25 ℃ and pH range of 6.0 to 8.5 with less than 30% change in its activity. The optimal temperature and pH were 15 ℃ and 7.5, respectively. Three peptides were determined by mass spectrometry which were shown to be not identical to other reported HAOs. Conclusion This is the first study to purify a low-temperature HAO from a heterotrophic nitrifier Acinetobecter sp. It differs from other reported HAOs in molecular mass and enzyme properties. The findings of the present study have suggested that the strain Y26 passes through a hydroxylamine-oxidizing process catalyzed by a low-temperature HAO for ammonium removal.

Isolation and characterization of heterotrophic nitrifying bacteria in MBR

The study presented the method for isolating the heterotrophic nitrifiers and the characterization of heterotrophic nitrification. Continuous tests via a membrane bioreactor (MBR) were operated under the controlled conditions to proliferate the nitrifiers. Heterotrophic nitrifying bacteria were isolated from the system in which the efficiency of total nitrogen(TN) removal was up to 80%. Since no autotrophic ammonium and nitrite oxidizers could be detected by fluorescence in situ hybridization(FISH), oxidized-N production was unlikely to be catalyzed by autotrophic nitrifiers during the heterotrophic nitrifiers' isolation in this study. The batch test results indicate that the isolated heterotrophic bacteria were able to nitrify. After 3 weeks incubation, the efficiencies of the COD removal by the three isolated bacterial strains B1, B2, and B3 were 52 6%, 71 7%, and 77 7%, respectively. The efficiencies of the TN removal by B1, B2, and B3 were 35 6%, 61 2% and 68 7%, respectively.