An ice-binding protein from an Antarctic sea ice bacterium

An Antarctic sea ice bacterium of the Gram-negative genus Colwellia, strain SLW05, produces an extracellular substance that changes the morphology of growing ice. The active substance was identified as a approximately 25-kDa protein that was purified through its affinity for ice. The full gene sequence was determined and was found to encode a 253-amino acid protein with a calculated molecular mass of 26,350 Da. The predicted amino acid sequence is similar to predicted sequences of ice-binding proteins recently found in two species of sea ice diatoms and a species of snow mold. A recombinant ice-binding protein showed ice-binding activity and ice recrystallization inhibition activity. The protein is much smaller than bacterial ice-nucleating proteins and antifreeze proteins that have been previously described. The function of the protein is unknown but it may act as an ice recrystallization inhibitor to protect membranes in the frozen state.     

Detection of <Emphasis Type="Italic">Erwinia amylovora</Emphasis> by novel chromosomal polymerase chain reaction primers

A new sensitive and specific method for the detection of Erwinia amylovora was developed. The method is based on the detection of a chromosomal DNA sequence specific for this bacterial species and enables detection of E. amylovora pathogenic strains, including recent isolates that lack plasmid pEA29 and thus cannot be detected by the previously popular PCR methods based on the detection of this plasmid. A species-specific random amplified polymorphic DNA (RAPD) marker was identified, cloned, and sequenced, and sequence characterized amplified region (SCAR) primers for specific PCR were developed. The E. amylovora specific sequence, 1269 bp long, was amplified in polymerase chain reaction and detected with electrophoresis in agarose gel stained with ethidium bromide. Amplification with other bacterial species did not produce any PCR product detectable by electrophoresis. Matching of the E. amylovora specific sequence to chromosomal DNA was confirmed by computer analysis of the E. amylovora genome. A consistent sensitivity limit of the method was 3 CFU/reaction, and in some cases it was possible to detect 0.6 CFU/reaction. Due to its high sensitivity and specificity, our method of E. amylovora detection is currently the most reliable, taking into account that the reliability of PCR methods based on plasmid pEA29 has been compromised by the isolation of pathogenic E. amylovora strains that lack this plasmid.     

海洋细菌E18菌株的生物学特性及其蓝紫色素稳定性的研究

从海泥中分离获得一株海洋细菌Ｅ18菌株,发现其具有稳定产生蓝紫色素的特性。对该菌的形态特征、培养特征及生理生化特征进行了研究。对该菌的分子鉴定结果表明,该菌为假交替单胞菌属细菌。萃取该菌的色素,并试验光、紫外线、热、pH、氧化剂及还原剂对该色素的影响,结果表明：该色素的最大吸收峰为579nm,紫外光对其稳定性无影响,但自然光对其有一定的消色作用。60℃～80℃温度范围对色素有一定的增色作用,而90℃以上高温可使色素消色。色素在pH3～9区域内稳定。色素对还原剂Na₂SO₃较为稳定, 而高浓度的氧化剂H₂O₂可使色素改变颜色。     

Relationship between mycobacteria and amoebae: ecological and epidemiological concerns

Since the discovery that Legionella pneumophila can survive and grow within free-living amoebae, there has been an increasing number of microbial species shown to have similar relationships. These include many bacterial species, fungi, other protozoa (e.g. Cryptosporidium) and viruses. Among bacteria, mycobacteria are of particular importance because of their role in human and animal infections. This review will consider the progress made in understanding the relationships between mycobacteria and amoebae, and their consequences in terms of ecology and epidemiology.     

三株光合细菌的毒性试验研究

以昆明小白鼠为实验动物,对类球红细菌Z1、沼泽红假单胞菌Z2及光合细菌分离株C2采用急性毒性试验、骨髓细胞微核试验和精子畸形试验进行安全性评定。结果表明,急性毒性试验:实验组小白鼠的一般状况、器官含水量、器官系数及血常规检查,与对照组差异均无显著性(P>0.05);骨髓细胞微核试验和精子畸形试验实验组与阴性对照组(生理盐水)差异均无显著性(P>0.05),与阳性对照组(环磷酰胺40 mg/kg)比较,骨髓细胞微核试验差异有极显著性(P<0.01),精子畸形率试验差异有显著性(P<0.05)。以上结果说明,三株光合细菌均不具有毒性。     

Biosynthesis of antibiotic prodiginines in the marine bacterium Hahella chejuensis KCTC 2396

AIMS: Hahella chejuensis KCTC 2396 produces red pigments, showing antibacterial and algicidal activities. The main red-coloured metabolite of the pigments was identified as antibiotic prodigiosin. With the expectation that the red pigments are a mixture of a series of close relatives, the aim of the present study is to detect new antibiotic prodigiosin analogues and to analyse the biosynthetic pattern for prodiginines in KCTC 2396. METHODS AND RESULTS: Except prodigiosin, the other constituents in the red pigments were confirmed as well-known dipyrrolyldipyrromethene prodigiosin, norprodigiosin, and undecylprodiginine. Additionally, four new prodigiosin analogues, each of which was distinguished from prodigiosin (C(5)), according to differences in alkyl chain length (C(3)-C(7)), were detected in small quantities by liquid chromatography mass spectrometry/mass spectrometry spectroscopy. Owing to the presence of a cytotoxic methoxy group, it is expected that all the new prodigiosin analogues are bioactive. CONCLUSIONS: Four characterized prodiginines, including prodigiosin and four new prodigiosin analogues are produced in different ratio in KCTC 2396. All of the prodiginines possess a common linear tripyrrolyl structure and a cytotoxic methoxy group. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows for the first time that KCTC 2396 is able to produce antibiotic prodigiosin, undecylprodiginine and new prodigiosin analogues in a mixture of pigments. It is also shown that KCTC 2396 possesses a novel system for the simultaneous production of multiple prodiginines in a single micro-organism.     

Identification of Tannerella forsythia antigens specifically expressed in patients with periodontal disease

Molecular pathogenesis of Tannerella forsythia, a putative periodontal pathogen, has not yet been adequately elucidated due to limited information on its virulence factors. Here, identification of in vivo expressed antigens of T. forsythia is reported using in vivo-induced antigen technology (IVIAT). Among 13 000 recombinant clones screened, 16 positive clones were identified that reacted reproducibly with sera obtained from patients with periodontal disease. DNA sequences from 12 of these in vivo-induced genes were determined. IVIAT-identified protein antigens of T. forsythia include: BspA, a well-defined virulence factor of T. forsythia; enzymes involved in housekeeping functions (tRNA synthetases, glycine hydroxymethyltransferase, and glucoside glucohydrolase); enzymes implicated in tissue destruction (dipeptidyl peptidase IV); a DNA mismatch repair protein; and putative outer membrane proteins of unknown function. The in vivo gene expression of these IVIAT-identified antigens was confirmed by a quantitative real-time PCR analysis. This is, to the best of the authors' knowledge, the first report using IVIAT in T. forsythia. It is anticipated that detailed analysis of the in vivo-induced genes identified by IVIAT in this study will lead to a better understanding of the molecular mechanisms mediating periodontal infection by T. forsythia.     

Nodule Symbiosis of Invasive <Emphasis Type="Italic">Mimosa pigra</Emphasis> in Australia and in Ancestral Habitats: A Comparative Analysis

Ninety isolates of root nodule bacteria from an invasive Mimosa pigra population in Australia were characterized by PCR assays and by sequencing of ribosomal genes. All isolates belonged to the same bacterial genus (Burkholderia) that predominates on M. pigra in its native geographic range in tropical America. However, the Australian Burkholderia strains represented several divergent lineages, none of which had a close relationship to currently known Burkholderia strains in American M. pigra populations. Inoculation of M. pigra with Australian strains resulted in equal or higher plant growth and nodule nitrogenase activity (measured by acetylene reduction assays) relative to outcomes with bacteria from M. pigra’s native geographic region. The main difference in symbiotic phenotype for bacteria from the two regions involved responses to an alternate Mimosa host species: Central American strains failed to fix nitrogen in association with Mimosa pudica, while most Australian Burkholderia isolates tested had high nodule nitrogenase activity in association with both Mimosa species. Invasive M. pigra populations in Australia have therefore acquired a diverse assemblage of nodule bacteria that are effective nitrogen-fixing symbionts, despite having a broader host range and a distant genetic relationship to bacterial strains found in the plant’s ancestral region.