[Identification and Nitrogen Removal Characteristics of a Heterotrophic Nitrification-Aerobic Denitrification Strain Isolated from Marine Environment].

A heterotrophic nitrification-aerobic denitrification strain named y5 was isolated from marine environment by traditional microbial isolation method using seawater as medium. It was identified as Klebsiella sp. based on the morphological, physiological and 16S rRNA sequence analysis. The experiment results showed that the optimal carbon resource was sodium citrate; the optimal pH was 7.0; and the optimal C/N was 17. The strain could use NH4Cl, NaNO2 and KNO3 as sole nitrogen source, and the removal efficiencies were77.07%, 64.14% and 100% after 36 hours, respectively. The removal efficiency reached 100% after 36 hours in the coexistence of NH4Cl, NaNO2 and KNO3. The results showed that the strain y5 had independent and efficient heterotrophic nitrification and aerobic denitrification activities in high salt wastewater.

[Isolation, Identification and Nitrogen Removal Characteristics of a Heterotrophic Nitrification-Aerobic Denitrification Strain y3 Isolated from Marine Environment].

A heterotrophic nitrification--aerobic denitrification bacterium named y3 was isolated from the sludge of Jiaozhou Bay using the enrichment medium with seawater as the matrix. It was identified as Pseudomonas sp. based on the morphological observation, physiological experiments and sequence analysis of 16S rRNA. The experiment results showed that the optimal carbon resource was sodium citrate, the optimal pH was 7.0, and the optimal C/N was 13. The strain could use NH4Cl, NaNO2 and KNO3 as sole nitrogen source, and the removal efficiencies were 98.69%, 78.38% and 72.95% within 20 hours, respectively. There was no nitrate and nitrite accumulation during the heterotrophic nitrification process. Within 20 hours, the nitrogen removal efficiencies were 99.56%, 99.75% and 99.41%, respectively, in the mixed system with NO3⁻-N: NO²â»-N of 2:1, 1:1 and 1:2. When the NH4⁺-N: NO3⁻-N ratios were 2: 1 , 1: 1 , 1: 2, the nitrogen removal efficiencies were all 100% . When the NH4⁺-N:NO2⁻-N ratios were 2:1,1:1,1:2, the nitrogen removal efficiencies were 90.43%, 92.79% and 99.96%, respectively. They were higher than those with single nitrogen source. As a result, strain y3 had good nitrogen removal performance in high saline wastewater treatment.

Carboxylicivirga linearis sp. nov., isolated from a sea cucumber culture pond.

A yellow-pigmented, Gram-stain-negative and facultatively anaerobic bacterium, designated FB218T, was isolated from a sediment sample collected from a sea cucumber culture pond in Rongcheng, China (36° 54' 36″ N 122° 14' 34″ E). Cells of strain FB218T were slender, gliding, catalase-positive and oxidase-negative. Optimal growth occurred at 30 °C, pH 6.5­7.0 and in medium containing 2­3 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain FB218T belonged to the genus Carboxylicivirga, family Marinilabiliaceae. The predominant fatty acids were iso-C15 : 0 and anteiso-C15 : 0. MK-7 was the main respiratory quinone. The major polar lipids of strain FB218T were two unidentified lipids and a phospholipid. The genomic DNA G+C content was 40.0 mol%. Based on the distinct phylogenetic position and the combination of physiological and phenotypic characteristics, strain FB218T represents a novel species of the genus Carboxylicivirga, for which the name Carboxylicivirga linearis sp. nov. is proposed. The type strain is FB218T ( = KCTC 42254T = MCCC 1H00106T). An emended description of the genus Carboxylicivirga is also provided.

Colwellia arctica sp. nov., isolated from Arctic marine sediment.

Strain 435(T), a catalase- and oxidase-positive, beige-pigmented, facultatively anaerobic and Gram-stain-negative marine bacterium, was isolated from marine sediment collected in the Arctic (8°21'629''E 72°8'827''N). The cells of the type strain are short- to curve-rods and able to grow at 4-25 °C, pH 6.0-9.0 and in the presence of 0.5-6.0 % (w/v) NaCl. It can reduce nitrate to nitrite. The predominant isoprenoid quinone was identified as Q-8 and the polar lipids are comprised of phosphatidylethanolamine and phosphatidylglycerol. The DNA G+C content is 38.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 435(T) belongs to the genus Colwellia. Strain 435(T) was found to exhibit 92.1-95.6 % 16S rRNA gene sequence similarities with other species of the genus Colwellia. The differential phenotypic properties, together with the phylogenetic and genetic distinctiveness indicated that strain 435(T) can be considered to represent a novel species, Colwellia arctica sp. nov., of the genus Colwellia. The type strain is 435(T) (=CICC 10860(T) = ATCC BAA-2609(T)).

Flaviflexus huanghaiensis gen. nov., sp. nov., an actinobacterium of the family Actinomycetaceae.

Strain H5(T) was isolated from a sediment sample collected from the coastal area of Qingdao, China. The cells were Gram-stain-positive, non-motile, straight or curved rods. The temperature range for growth was 20-37 °C and the pH for growth ranged from 6.5 to 9.0, with optimum growth occurring in the temperature range 28-30 °C and pH range 7.5-8.0. Growth occurred in the presence of 0-6% (w/v) NaCl (optimum, 0-2%). Strain H5(T) had MK-9, MK-9(H2) and MK-9(H4) as the major menaquinones and C18:1ω9c, C16:0, C14:0, C18:0 and C16:1ω9c as major fatty acids. The cell-wall peptidoglycan type was A5α l-Lys-l-Ala-l-Lys-d-Glu. The major polar lipids were phosphatidylglycerol (PG), an unknown phospholipid (PL1) and two unknown phosphoglycolipids (PGL1, PGL2). An unknown phospholipid (PL2) and two unknown glycolipids (GL1, GL2) were present in moderate to minor amounts in the polar lipid profile. The genomic DNA G+C content was 61.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain H5(T) represents a novel lineage in the family Actinomycetaceae. On the basis of phenotypic, physiological and molecular characteristics, it is proposed that the novel isolate should be classified as a novel species in a new genus: Flaviflexus huanghaiensis gen. nov., sp. nov., with strain H5(T) ( = DSM 24315(T) =CICC 10486(T)) as the type strain of the type species.

Preoperative inhalation of milrinone attenuates inflammation in patients undergoing cardiac surgery with cardiopulmonary bypass.

OBJECTIVES: The purpose of this study was to evaluate the effect of preoperative inhalation of milrinone on cardiopulmonary bypass (CPB)-related inflammation. SUBJECTS AND METHODS: A total of 30 patients undergoing cardiac surgery were recruited and randomized for preoperative inhalation of milrinone (Mil group) or normal saline (NS group), respectively. Each group had 15 patients. Their hemodynamic parameters were measured and blood samples were obtained longitudinally. The levels of serum interleukin (IL-6), tumor necrosis factor-α (TNF-α), and matrix metalloproteinase (MMP)-9 were determined by ELISA. RESULTS: The levels of serum IL-6, TNF-α, and MMP-9 significantly increased at the end of cardiac surgery and remained high for 24 h in both groups of patients. However, the levels of proinflammatory cytokines at the end of cardiac surgery in the Mil group of patients were significantly lower than those of the NS group of patients. CONCLUSIONS: Our data indicated that preoperative inhalation of milrinone significantly mitigated CPB-related inflammation.

[Effect of cardiopulmonary bypass on the expression of survivin in neutrophils].

OBJECTIVE: To investigate the effects of cardiopulmonary bypass on neutrophils apoptosis and the expression of survivin. METHODS: Ten patients who scheduled for cardiac surgery under cardiopulmonary bypass were recruited as study group and 10 healthy volunteers as control. Blood samples were obtained before operation, at the end of surgery, and at 24 hours postoperatively. Neutrophils were isolated by density gradient centrifugation and its apoptosis were evaluated by fluorescent microscope and flow cytometry. The expression of survivin protein was examined by Western blotting analysis. Expression level of survivin mRNA was detected by RT-PCR. RESULTS: The apoptotic rate of neutrophils decreased significantly at the end of surgery (P < 0.01), and was still lower at 24 hours postoperatively than before operation (P < 0.05). The expression ratios of survivin protein and mRNA were increased at the end of surgery (P < 0.01), and decreased gently at 24 hours postoperatively but was still higher than before operation (P < 0.05). CONCLUSION: Cardiopulmonary bypass could inhibit neutrophils apoptosis and increase the expression of survivin. The decrease of neutrophils apoptosis was correlated with high expression of survivin.

Molecular cloning and tissue expression patterns of a small conductance calcium-activated potassium channel gene in turbot (Scophthalmus maximus L.).

Calcium-activated potassium channels on plasma membrane enable potassium influx into the cell with ensuing changes in plasma membrane potential and consequent effects on cellular metabolic functions. Recently, this potassium channel was reported to regulate the cellular responses of mammalian immune cells. We have postulated the presence of such a channel in fish immune cells and its potential role in immunoregulation in fish. Employing specific primers and RNA template, we cloned a segment of a novel gene from turbot blood sample and subsequently obtained a full cDNA sequence using RACE approaches. Bioinformatic analysis revealed structural and phylogenetic characteristics of a novel small conductance calcium-activated potassium channel gene, we called TSKCa, which exhibits homologous domains to other species particularly in the transmembrane regions. Full-length TSKCa cDNA is 1698 bp with a 1632 bp open reading frame encoding a protein of 544 amino acids. TSKCa gene is expressed in majority of the tested organs and tissues of turbot. To assess the postulated immune function of TSKCa, we infected turbot with the pathogen Vibrio anguillarum. Here, semi-quantitative RT-PCR analysis demonstrated increased mRNA expression of TSKCa in head kidney, spleen and blood, indicating an important role of TSKCa in these organ tissues that mediate the immune defense response of turbot. In contrast, there was less change in expression in the turbot intestines and liver which were less implicated in the immune response in present study.

[Effect of spectra on growth of chlorella and isochrysis].

Focusing on the speed and efficiency, the effects of different spectra on the growth of chlorella vulgaris and isochrysis galbana Parke 8701 were investigated by using monochromatic LED (light-emitting diode) and fluorescent lamp as light sources. It was concluded that continuous spectra accelerate the top-growth-rate, blue light has the best efficiency, and the combination of them can obtain a good balance of speed and efficiency. For the purpose of measuring spectra as a parameter of irradiation quantitatively, spectra-absorbability-coefficient defined as the quanta-absorbability- efficiency of spectra for algae was calculated by means of absorption spectra of algae and emission spectra of light sources. Compared with the experimental results the coefficients of different light sources have a positive correlation to their efficiency for growth, so the coefficient can be used to elementarily quantify the relation between the spectra and the efficiency for growth.