Antibacterial activities of the leaf extracts of Eugenia uniflora Linn. (Synonym Stenocalyx michelli Linn.) Myrtaceae

The aqueous, organic, and volatile oil extracts of leaves of Eugenia uniflora Linn. Family Myrtaceae were investigated for antibacterial properties using agar dilution techniques. The aqueous extract was the most active against the organisms compared to the organic and volatile oil extracts. The extracts were found to inhibit Gram positive Staphylococcus aureus and Bacillus subtilis and Gram negative Escherichia coli and Shigella dysentcriae. Pseudomonas aeruginosa, Klebsiella pneumoniac, and Salmonella typhi were not inhibited.     

我国37年来细菌性痢疾菌群分布的分析

本文复习了我国37年(1949~1985)来自各地区报道的155 122株痢疾杆菌。分析菌群分布表明,平均仍以B亚群所占比例最高(80.5%),但八十年代以来有所下降(68.1%)。B亚群中流行亚群是2a与lb;但1a与2b比例有增加趋势。西北和中南地区A亚群比例近来明显增高,提示有关地区环境污染严重,饮水卫生差,亦可能与细菌毒力变异有关,防治中需特别注意。B:D亚群比值以华东地区最低,该比值同经济文化发达水平有一定关系。     

Comparison of the EUCAST-AFST broth dilution method with the CLSI reference broth dilution method (M38-A) for susceptibility testing of posaconazole and voriconazole against Aspergillus spp.

The susceptibilities of 40 clinical isolates of Aspergillus spp. (Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus terreus) were determined for posaconazole and voriconazole by the CLSI M38-A and EUCAST-AFST broth dilution methods. Where a discrepancy was observed between the methods, the EUCAST method tended to give higher MIC values. Overall, the level of agreement was 92.5% and the intra-class correlation coefficient was > 0.90.     

福氏志贺菌毒力蛋白IpaC的表达与纯化

目的表达和纯化福氏志贺菌毒力蛋白IpaC,研究福氏志贺菌的致病机制。方法将含有ipaC基因的pET32a-i-paC表达质粒载体转化大肠杆菌BL21(λDE3),在异丙基硫代-β-D半乳糖苷(IPTG)诱导下表达,对诱导后的表达产物进行SDS-PAGE鉴定,并采用QIA expressionistTM蛋白纯化系统来纯化目的蛋白。结果诱导后的表达产物经SDS-PAGE发现有一相对分子质量约为63 000的条带,其含量约占总蛋白量的11%,以350 mmol/L咪唑洗脱液洗脱,目的蛋白纯度可达90%以上。结论pET32a-ipaC重组表达质粒转入大肠杆菌后,可稳定、高效地表达目的蛋白,QIA-expressionistTM蛋白纯化系统是一种简便、高效的纯化系统,可获得高纯度的目的蛋白。     

Morphology of temperate bacteriophage SfV and characterisation of the DNA packaging and capsid genes: the structural genes evolved from two different phage families

The entire genome of SfV, a temperate serotype-converting bacteriophage of Shigella flexneri, has recently been sequenced (Allison, G.E., Angeles, D., Tran-Dinh, N., Verma, N.K. 2002, J. Bacteriol. 184, 1974-1987). Based on the sequence analysis, we further characterised the SfV virion structure and morphogenesis. Electron microscopy indicated that SfV belongs to the Myoviridae morphology family. Analysis of the proteins encoded by orf1, orf2, and orf3 revealed that they were homologous to small and large terminase subunits, and portal proteins, respectively; the protein encoded by orf5 showed homology to capsid proteins. Western immunoblot of the phage with anti-SfV sera revealed two antigenic proteins, and the N-terminal amino acid sequence of the 32-kDa protein corresponded to amino acids 116 to 125 of the ORF5 protein, suggesting that the capsid may be processed. Functional analysis of orf4 showed that it encodes the phage capsid protease. The proteins encoded by orfs1, 2, 3, 4, and 5 are homologous to similar proteins in the Siphoviridae phage family of both gram-positive and gram-negative origin. The capsid and morphogenesis genes are upstream and adjacent to the genes encoding Myoviridae (Mu-like) tail proteins. The organisation of the structural genes of SfV is therefore unique as the head and tail genes originate from different morphology groups.     

Detection of Brucella melitensis by the BacT/Alert automated system and Brucella broth culture

This study was conducted to evaluate the ability of the BacT/Alert automated blood culture system to detect Brucella spp. in comparison with traditional Brucella broth culture. Overall, 100 (50 bone marrow and 50 blood samples) paired cultures were obtained, and 59 were positive by at least one method. The Brucella broth culture method detected all 59 positive cultures (100%), and the BacT/Alert system detected 30 (50.8%) ( P  < 0.05). The mean detection times for B. melitensis were 4.5 days in the BacT/Alert system and 5 days in Brucella broth culture ( P  > 0.05). There is no significant difference between the two methods with respect to growth time of the microorganism, but Brucella broth culture is more sensitive than the BacT/Alert system.     

Molecular characterisation of extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella spp. isolates at a tertiary-care centre in Lebanon

The prevalence of bla _CTX-M, bla _TEM and bla _SHV genes among extended-spectrum β-lactamase (ESBL)-producing clinical isolates of Escherichia coli ( n  = 50) and Klebsiella spp. ( n  = 50) from Lebanon was 96%, 57% and 67%, and 40%, 82% and 84%, respectively. Genotyping revealed that the clonal diversity was unrelated to the presence of bla genes. Sequence analysis of 16 selected isolates identified the bla _CTX-M-15, bla _TEM-1, bla _OXA-1 and six bla _SHV genes, as well as the gene encoding the quinolone-modifying enzyme AAC(6')-Ib-cr. The genes encoding CTX-M-15 and AAC(6')-Ib-cr were carried on a 90-kb plasmid of the pC15-1a or pCTX-15 type, which transferred both ESBL production and quinolone resistance from donors to transconjugants.     

Effects of SITS on urate transport by isolated,perfused snake renal tubules

Effects of an inhibitor of membrane anion-exchange transport processes, 4-acetamido-4-isothiocyano-2,2-disulfonic stilbene (SITS), on urate transport by isolated, perfused snake (Thamnophis spp.) proximal renal tubules were studied. SITS (10^–4 mol/l) in the luminal perfusate had absolutely no effect on net urate secretion (J _urate ^net ) or on net fluid absorption (J _v). This observation is compatible with other data that give no support to the concept of a mediated transport step for urate from the cells to the lumen. SITS (10^–4 mol/l) in the bathing medium reversibly inhibitedJ _urate ^net without affectingJ _v. At the time of maximum inhibition ofJ _urate ^net , the concentration of urate in the cell water was increased and the apparent permeability of the luminal membrane to urate was decreased, but the urate efflux across the peritubular membrane and the apparent permeability of the peritubular membrane to urate were unchanged. There was no evidence of significant intracellular binding or trapping of urate. Although an increase in the initial rate of urate transport into the cells across the peritubular membrane could not be demonstrated conclusively in nonperfused tubules, the results still suggest that SITS in the bathing medium may inhibitJ _urate ^net by inhibiting urate movement from the cells to the lumen while actually enhancing transport from the bathing medium into the cells.     

Ribosomal DNA internal transcribed spacer sequences do not support the species status of Ampelomyces quisqualis, a hyperparasite of powdery mildew fungi

Phylogenetic relationships among Ampelomyces isolates, pycnidial hyperparasites and biological control agents of powdery mildews, were inferred from internal transcribed spacer (ITS) sequences of the ribosomal DNA (rDNA). Currently, these hyperparasites are considered to be a single species, A. quisqualis, despite observed morphological and cultural differences. Ten Ampelomyces isolates, representing seven previously defined ITS RFLP groups, were sequenced and analyzed. Sequence-divergence values among isolates belonging to different RFLP groups ranged from 4.3 to 22.4%, suggesting that these isolates may represent different taxa. When Ampelomyces ITS sequences were analyzed by cladistic methods with the sequences of other ascomycetous fungi, they formed two lineages in the Dothideales. Slow-growing Ampelomyces isolates formed a clade with Leptosphaeria microscopica and L. nodorum, whereas fast-growing Ampelomyces isolates formed a clade with Epicoccum nigrum. Sequence-divergence values between these two clades ranged from 17.3 to 22.4%, suggesting that the taxa in the two clades are not closely related and possibly not congeneric. The data presented here indicate that the identification of `A. quisqualis' isolates used in biological control experiments should be re-evaluated. Received: 10 March 1997 / Accepted: 13 February 1998     

Genetic relatedness and antimicrobial resistance determinants among clinical isolates of enterococci from Cuba

This study describes the genetic relationships and antimicrobial resistance determinants found among 99 clinical isolates of enterococci from 15 different hospitals in Cuba. Pulsed-field gel electrophoresis SmaI analysis demonstrated a high degree of genetic diversity. A limited number of multiresistant Enterococcus faecalis clones, showing resistance to three or more families of antimicrobial agents, were detected simultaneously in different institutions, suggesting inter-hospital circulation of selected clones, and/or selection of particular clones following their introduction into the hospital environment. Antimicrobial resistance determinants, including erm(B), aac(6')-aph(2'), aph(3'), ant(6), vanB (E. faecalis) and vanA (Enterococcus faecium) were detected by PCR in various isolates.