AhaP,A Quorum Quenching Acylase from Psychrobacter sp. M9-54-1 That Attenuates Pseudomonas aeruginosa and Vibrio coralliilyticus Virulence

Although Psychrobacter strain M9-54-1 had been previously isolated from the microbiota of holothurians and shown to degrade quorum sensing (QS) signal molecules C6 and C10-homoserine lactone (HSL), little was known about the gene responsible for this activity. In this study, we determined the whole genome sequence of this strain and found that the full 16S rRNA sequence shares 99.78–99.66% identity with Psychrobacter pulmonis CECT 5989^T and P. faecalis ISO-46^T. M9-54-1, evaluated using the agar well diffusion assay method, showed high quorum quenching (QQ) activity against a wide range of synthetic N-acylhomoserine lactone (AHLs) at 4, 15, and 28 °C. High-performance liquid chromatography-mass-spectrometry (HPLC-MS) confirmed that QQ activity was due to an AHL-acylase. The gene encoding for QQ activity in strain M9-54-1 was identified from its genome sequence whose gene product was named AhaP. Purified AhaP degraded substituted and unsubstituted AHLs from C4- to C14-HSL. Furthermore, heterologous expression of ahaP in the opportunistic pathogen Pseudomonas aeruginosa PAO1 reduced the expression of the QS-controlled gene lecA, encoding for a cytotoxic galactophilic lectin and swarming motility protein. Strain M9-54-1 also reduced brine shrimp mortality caused by Vibrio coralliilyticus VibC-Oc-193, showing potential as a biocontrol agent in aquaculture.     

Synergistic and antagonistic hemolytic activities of bacteria of genus Arcanobacterium and CAMP-like hemolysis of Arcanobacterium phocae and Arcanobacterium haemolyticum with Psychrobacter phenylpyruvicus

A total of 57 bacteria representing eight species of genus Arcanobacterium (A.) were investigated for hemolytic properties on blood agar containing sheep and rabbit blood and for CAMP-like reactions. An enhanced hemolysis on blood agar containing rabbit blood compared to sheep blood could be observed for A. haemolyticum, less pronounced for A. hippocoleae and A. pluranimalium.A synergistic hemolytic reaction with staphylococcal β-hemolysin appeared to be constantly visible for A. hippocoleae, A. pluranimalium and A. pyogenes, with Streptococcus agalactiae for A. phocae and A. haemolyticum, with Rhodococcus equi for A. phocae, A. haemolyticum, A. pluranimalium and A. pyogenes and with A. haemolyticum for A. hippocoleae, A. pluranimalium and A. pyogenes, respectively. A reverse CAMP-reaction in the zone of staphylococcal β-hemolysin could be observed for A. phocae and A. haemolyticum. In addition, a novel CAMP-like reaction could be noted between Psychrobacter phenylpyruvicus, identified by 16S rDNA sequencing, and A. phocae and A. haemolyticum. These synergistic or antagonistic hemolytic properties could possibly be used as additional criteria for identification of bacteria of genus Arcanobacterium.     

Structural Investigation of the Oligosaccharide Portion Isolated from the Lipooligosaccharide of the Permafrost Psychrophile Psychrobacter arcticus 273-4

Psychrophilic microorganisms have successfully colonized all permanently cold environments from the deep sea to mountain and polar regions. The ability of an organism to survive and grow in cryoenviroments depends on a number of adaptive strategies aimed at maintaining vital cellular functions at subzero temperatures, which include the structural modifications of the membrane. To understand the role of the membrane in the adaptation, it is necessary to characterize the cell-wall components, such as the lipopolysaccharides, that represent the major constituent of the outer membrane. The aim of this study was to investigate the structure of the carbohydrate backbone of the lipooligosaccharide (LOS) isolated from the cold-adapted Psychrobacter arcticus 273-4. The strain, isolated from a 20,000-to-30,000-year-old continuously frozen permafrost in Siberia, was cultivated at 4 °C. The LOS was isolated from dry cells and analyzed by means of chemical methods. In particular, it was degraded either by mild acid hydrolysis or by hydrazinolysis and investigated in detail by ¹H and ¹³C NMR spectroscopy and by ESI FT-ICR mass spectrometry. The oligosaccharide was characterized by the substitution of the heptose residue, usually linked to Kdo in the inner core, with a glucose, and for the unusual presence of N-acetylmuramic acid.     

Effect of dietary administration of Psychrobacter sp. on the growth,feed utilization,digestive enzymes and immune responses of grouper Epinephelus coioides

A 60‐day feeding trial was conducted to assess the effect of incorporating dietary Psychrobacter sp., one predominant bacterium with antagonistic activity in the gut of fast‐growing grouper Epinephelus coioides, on the growth, feed utilization, digestive enzymes and immune responses of E. coioides at regular monthly intervals. The fish were fed with control diet (without probiont) or probiotic diet supplemented with 1.0 × 10⁸ CFU g^?1Psychrobacter sp. No significant improvement in weight gain (WG) and specific growth ratio (SGR) were observed in the probiotic group, but a significant improvement in feed conversion ratio (FCR) was observed in fish fed the probiotic diet for 60 days. Although not statistically significant, enhanced hepatopancreatic protease and lipase activities and intestinal amylase activity were observed in probiotic group at day 60. A slightly increase in phagocytic activity and phagocytic index was observed in fish fed probiotic diet for 60 days. Serum lysozyme activity showed no significant differences between the probiotic group and the control group during the whole experiment period, while an increasing trend of superoxide dismutase (SOD) activity in the probiotic group was observed. Serum complement component 3 (C3) level of the probiotic group was higher than that of control in the whole feeding period, while complement component 4 (C4) level of the probiotic group was significantly higher than that of the control at day 60. The results of the current study firstly demonstrated potential for Psychrobacter sp. as a novel probiont to improve feed utilization, digestive enzymes and innate immunity in E. coioides.     

裙带菜孢子体溃烂病原鉴定及病因分析

为探究大连市龙王塘和铁山增养殖海域养殖裙带菜Undaria pinnatifida孢子体出现溃烂的原因,利用细菌分离及分子生物学方法,从海域养殖裙带菜溃烂的孢子体中分离纯化得到2株优势菌株(编号分别为LH43和LQ2),通过人工感染试验证实,分离菌株能够使裙带菜孢子体出现溃烂并分解藻体,且与自然发病相似,LH43较LQ...     

南极菌B-3胞外寡聚糖对黄瓜的诱导抗病作用

[目的]研究南极菌B-3对黄瓜的诱导抗病作用。[方法]以黄瓜幼苗为材料,研究南极菌B-3胞外寡聚糖的分子量、单糖组成,及对黄瓜叶中防御酶的诱导作用与黄瓜白粉病的诱导抗病性。[结果]B-3胞外寡聚糖经高效凝胶渗透色谱测定其分子量为2112D,气相色谱分析由甘露糖和葡萄糖2种单糖组成;0.3%、0.5%和0.8%的B-3胞外多糖分别对黄瓜幼苗喷施不同时间后,黄瓜体内的几丁质酶,β-1,3-葡聚糖酶,苯丙氨酸解氨酶（PAL）,过氧化物酶（POD）的活性均较对照有不同程度的提高,且与0.5%的壳聚糖具有相似的诱导效果;同时,0.5%的B-3多糖能够显著地降低黄瓜白粉病的病情指数,防效达24.49%。[结论]B-3寡聚糖有望开发为新型抗病诱导子。