- Volume 46, Issue 4, 1997
Volume 46, Issue 4, 1997
- Editorial
-
- Epidemiological Typing
-
-
-
Genome organisation of Staphylococcus aureus isolates from different populations
More LessIsolates from three different Staphylococcus aureus populations were examined for restriction fragment length polymorphisms (RFLPs) of total DNA digested with the endonucleasc SmaI. The populations were: community S. aureus isolates collected at random from healthy individuals (38 isolates): methicillin-resistant S. aureus (MRSA) type strains involved in separate outbreaks of infection in Melbourne (1982) and Canberra (1990) (two isolates); and a collection of clinical methicillin-sensitive S. aureus (MSSA) causing hospital infection (20 isolates). RFLPs with CspI and SmaI and hybridisation analyses of both, showed that the community and the MSSA isolates were not genetically closely related, and, accordingly, they could not be grouped into clusters as seen with the MRSA types. However, a few MSSA isolates were found to be closely related to each other and appeared to be similar to the standard strain S. aureus 8325–4 and to some MRSA types. Although there was substantial variability between the three groups, physical mapping with genomic DNA fragments from the standard strain S. aureus 8325-4 to probe large fragments generated with CspI and SmaI from the chromosomes of selected community and MRSA isolates, demonstrated a well conserved genome organisation between representative isolates from the three groups.
-
-
-
-
A three-way ribotyping scheme for Salmonella serotype Typhimurium and its usefulness for phylogenetic and epidemiological purposes
More LessRibotyping of Salmonella serotype Typhimurium strains was optimised as a tool for epidemiological and phylogenetic purposes. Of five restriction endonucleases evaluated on a series of 84 isolates, HincII, Sal/I and PvuII were the most useful, generating 13, 9 and 9 ribotypes with 17, 11 and 18 polymorphic restriction sites, and attaining a discrimination index (DI) of 0.81, 0.53 and 0.59, respectively. The combination of results from tests with the three enzymes provided further discrimination (19 ribotypes, DI = 0.84). It proved useful for clonal analysis, defining 19 clonal lines with a remarkable degree of genetic heterogeneity, that were grouped into two major clusters (including 12 and 7 lines, respectively) at a significance level of 0.65. When the attributes of this system were compared with those of phage typing, it was found that ribotyping showed higher typability and sensitivity, supporting its use as an appropriate molecular method. In tracing the molecular epidemiology of Typhimurium strains in Asturias, six lines were found that could be considered endemic and were represented by organisms implicated in salmonellosis throughout the period of study; another four lines included organisms isolated from meat, water or both.
-
-
-
Direct comparison of two commercially available computer programs for analysing DNA fingerprinting gels
More LessRandomly amplified polymorphic DNA (RAPD) fingerprints were generated with M13 and DAF4 primers for 25 isolates of Acinetobacter spp. obtained from 16 different hospitals situated in 12 countries. The overall robustness of the algorithms and the reproducibility of the cluster analysis results generated by two commercially available computer programs (GelCompar and DENDRON) for analysing DNA fingerprinting gels were tested by examining the same set of fingerprinting data independently in two laboratories with the different software packages. Both programs were efficient at recognising and grouping strains with closely similar RAPD fingerprints, i.e., strains which might be expected to have a close epidemiological or evolutionary relationship. However, the relationships suggested for less closely related strains showed considerable variation in terms of the overall similarity or percentage correlation values suggested by the programs. It was concluded that both programs were useful tools for indicating close genotypic relationships between individual strains, but that epidemiological conclusions based on the similarity or correlation values (or the dendrograms derived from them) obtained for less closely related strains should be treated with considerable caution.
-
- Technical Note
-
-
-
Growth of Helicobacter pylori in candle jars
More LessCulture of Helicobacter pylori requires micro-aerobic conditions which are usually generated by a commercial gas-generator system. However, the commercial gas-generator pack is not available in most hospitals in developing countries. The present study showed that H. pylori grew well in a candle jar system that provided micro-aerobic condition; this gave similar cultural results to a commercial gas-generator system. Therefore, the candle jar system can be used for H. pylori micro-aerobic culture in developing countries where commercial systems are not available.
-
-
- Bacterial Pathogenicity
-
-
-
Characteristics of toxicity and haemorrhagic toxin produced by Clostridium sporogenes in various animals and cultured cells
More LessThe toxic effects of the haemorrhagic toxin of Clostridium sporogenes were studied in mice, rats, guinea-pigs and rabbits, and in various cultured cells. In rabbits, but not in the other animals, intradermal injection with crude toxin and its injection into a ligated intestinal loop caused haemorrhage in both the skin and intestinal wall. Intraperitoneal (i.p.) injection of crude toxin similarly caused death only of rabbits, with marked haemorrhage in the serous surface of kidney, intestines, liver, spleen, mesentery and diaphragm. Histological examination of the rabbits killed after i.p. inoculation revealed leakage of blood into a space beneath the serous membranes of parenchymatous organs in the peritoneal cavity and within the loose connective tissues in the mesentery and diaphragm. Cytotoxicity of partially purified haemorrhagic toxin in vitro was noted with rabbit aorta endothelial cells, human skin capillary vein endothelial cells and bovine pulmonary artery endothelial cells, but not with Chinese hamster ovary cells, Vero cells, human epitheloid carcinoma cells, human colon carcinoma cells (T84) and human colon adenoearcinoma cells (Caco 2). The results suggest that the haemorrhagic toxin of C. sporogenes exerts its effects in rabbits but not in mice, rats or guinea-pigs, through direct action on endothelial cells.
-
-
-
-
Characterisation of the leukotoxin produced by different strains of Pasteurella haemolytica
More LessPasteurella haemolytica isolates from cattle and sheep, including representatives of different serotypes and untypable strains, were examined for leukotoxin (Lkt) production at the end of the log phase of growth in brain heart infusion broth. There were marked differences in leukotoxic activity in culture supernate samples, as measured by chemiluminescence-inhibition assays with bovine and ovine neutrophils, even between strains of the same serotype. There was also some variation in the amount and mol. wt of the Lkt protein produced by different strains, as judged by SDS-PAGE, immunoblotting and ELISA. Some strains produced normal amounts of Lkt protein which had only low leukotoxic activity. Most strains produced Lkt of 105 kDa whereas four strains produced a higher mol. wt form of c. 108 kDa, including two of the five serotype A2 strains examined. Thus, the P. haemolytica isolates showed considerable heterogeneity in terms of leukotoxin production, mol. wt and activity, even within a given serotype.
-
-
-
Vitronectin may mediate staphylococcal adhesion to polymer surfaces in perfusing human cerebrospinal fluid
More LessProsthetic devices are frequently used for temporary or permanent drainage of cerebrospinal fluid (CSF), i.e., ventricular catheters with or without external monitoring devices and shunts. Infections constitute a serious complication in the use of biomaterials in contact with CSF; coagulase-negative staphylococci (CNS) are the most common aetiological agents. In the present study, polyvinylchloride (PVC) and PVC with endpoint-attached heparin were exposed to human CSF under perfusion to mimic conditions in vivo. Adhesion of strains of CNS isolated from patients with or without biomaterial-associated infection was determined: (i) after pre-incubation with fibronectin (Fn) or vitronectin (Vn) to block bacterial surface binding structures; and (ii) after pre-incubation of biomaterials with antibodies to Fn or Vn to block exposure of bacteria-binding domains on these host proteins. Pre-incubation of bacterial cells with Vn significantly reduced subsequent adhesion to polystyrene precoated with Vn 0.5 μg/ well. When PVC pre-exposed to CSF was incubated with antibodies to Vn, subsequent bacterial adhesion of a Vn-binding strain, S. epidermidis 5703, was significantly reduced. The study shows that Vn may mediate adhesion of CNS in the presence of CSF. However, strains retrieved from biomaterials did not express binding of Vn or Fn to a higher extent than non-biomaterial-associated strains. Expression of heparin binding under static conditions did not correlate with staphylococcal adhesion to heparinised polymers under perfusion with CSF. The extent of adhesion of staphylococci to heparinised PVC was either reduced or the same as to unheparinised PVC.
-
- Molecular Diagnosis
-
-
-
Strategy for detection and identification of bacteria based on 16S rRNA genes in suspected cases of Whipple’s disease
More LessThe 16S ribosomal RNA (rRNA) gene of the phylogenetic subdivision containing gram-positive bacteria with a high G + C content was detected specifically in clinical specimens from patients suspected of having Whipple’s disease. The primary structure of 16S rDNA amplified from clinical samples was determined by cloning and sequencing. Two sorts of sequences were identified: one corresponded exactly to the rRNA sequence of Tropheryma whippelii (GenBank accession no. M87484) while the other was related to that of members of the genus Corynebacterium. No sequence related to Mycobacterium spp. or Rhodococcus equi was observed. Exhaustive examination of negative specimens with broad-range eubacterial primers detected one sequence related to Enterobacter-iaceae and another related to Enterococcus spp. To speed identification of T. whippelii, a nested amplification method was devised. A first amplification specific for the gram-positive bacteria subdivision was performed, followed by a second amplification with T. whippelii-specific primers. The amplified T. whippelii product was checked by digestion with Ava II, StuI, and PstI endonucleases. These techniques were applied to DNA extracted from seven intestinal biopsy samples, two cerebrospinal fluid samples and one articular fluid from patients suspected of having Whipple’s disease. T. whippelii 16S rDNA was found in two of the biopsy samples, one of the cerebrospinal fluid samples and in the articular fluid.
-
-
-
-
Differentiation of strains of Mycoplasma fermentans from various sources by pyrolysis mass spectrometry
More LessMycoplasma fermentans has attracted much interest both as a cofactor for the progression of AIDS and as a pathogenic agent in non-AIDS related diseases. Previous studies with serological and genetic techniques suggest that M. fermentans represents a homogeneous group of organisms, with no significant differences identified among the strains examined. In this study, 25 cultures of M. fermentans, including isolates from human sources and tissue culture cells, were compared by pyrolysis mass spectrometry (PMS). It was possible to distinguish the‘type’ strain PG-18 from an AIDS-associated M. fermentans strain‘incognitus’ by this technique. PMS was also able to differentiate laboratory-induced aminoglycoside-resistant variants from their fully susceptible parents. Four AIDS-associated isolates were distinguished from each other, whilst five European cell culture isolates were shown to be closely related, as were six M. fermentans isolates from an outbreak of acute respiratory infection in Canada. PMS has proved useful in distinguishing isolates of M. fermentans, providing epidemiological data. In addition, PMS may help in determining the likely origin of a given isolate, and in the future may be of use in assessing the role of this micro-organism in human disease.
-
- Mycology
-
-
-
GM-CSF-modulated phagocytosis of Trichosporon beigelii by human neutrophils
More LessTrichosporon beigelii has emerged as a lethal opportunist pathogen in granulocytopenic and corticosteroid-treated patients. Little is known of the host defence mechanisms against this yeast. The interaction between human neutrophils and serum-opsonised T. beigelii and the effect of GM-CSF on binding and ingestion of the yeast by neutophils were investigated by a microscopic analysis of neutrophil monolayers stained with FITC-Concanavalin A. Positive staining with FITC-Concanavalin A distinguished between intracellular and extracellular yeast cells. Binding of T. beigelii to neutrophils was an energy- and complement-dependent process involving movement of actin in the neutrophil cytoskeleton. The mean percentage binding of T. beigelii was 37.5% and the mean binding index (BI) was 1.30 whereas the mean percentage ingestion was 3.5% and the mean phagocytic index (PI) was 1.34. GM-CSF increased percentage ingestion of T. beigelli from 2.8% to 30.5% and the PI was increased from 1.3 to 1.86. The percentage binding was 36.8% and the mean BI was 1.3 in control experiments compared with 49.3% and 1.6, respectively, in the presence of GM-CSF. In conclusion, GM-CSF significantly increased percentage ingestion of opsonised T. beigelii by neutrophils, but its effect on percentage binding of the yeast was not statistically significant.
-
-
-
-
Scanning electron microscopy of the development of structured aerial mycelia and satellite colonies of phenotypically switched Candida albicans
More LessCandida albicans is an asexual diploid fungus that can express high frequency phenotypic switching. Switched variants can develop structured aerial mycelia (SAM) and cultures that have been grown for a protracted period exhibit satellite colonies. This study investigated the development of both SAM and satellite colonies by means of a freeze-drying technique and scanning electron microscopy. The results show that SAM may develop due to hyphae developing in the colony at the base of the structures. The development of satellite colonies is a result of cells that have grown deep into the agar re-emerging on to the agar/air surface and producing a new colony. Although both SAM and satellite colonies are often seen together on mature colonies, their cause and development are different.
-
- Serological Diagnosis
-
-
-
Diagnostic potential of a serological assay for the diagnosis of ulcerans disease based on the putative Mycobacterium ulcerans toxin
More LessMycobacterium ulcerans infection is an important and potentially disfiguring disease of man. A rapid diagnostic assay for detection of this organism is required urgently. Serological assays require a species-specific protein to ensure a high level of specificity and thus reduce the occurrence of false positive results. As M. ulcerans had been reported to produce a unique cytotoxin, it was thought that this would provide an ideal antigen on which to base a serological assay for detection of M. ulcerans during infection. Crude culture filtrates, prepared by previously documented methods, were assayed for toxic activity by in-vitro cytotoxicity assays and in-vivo mouse footpad assays. To evaluate the uniqueness of the cytotoxic factor, other species of mycobacteria were also assayed. Analysis of these assays showed that similar biological activity is present in various other mycobacterial species. Furthermore, it was possible to neutralise this activity in all species tested with a polyclonal antiserum raised against M. ulcerans. As the cytotoxic factor was found not to be specific to M. ulcerans, it is unlikely that a serological assay based on such a molecule will be of use.
-
-
Volumes and issues
-
Volume 73 (2024)
-
Volume 72 (2023 - 2024)
-
Volume 71 (2022)
-
Volume 70 (2021)
-
Volume 69 (2020)
-
Volume 68 (2019)
-
Volume 67 (2018)
-
Volume 66 (2017)
-
Volume 65 (2016)
-
Volume 64 (2015)
-
Volume 63 (2014)
-
Volume 62 (2013)
-
Volume 61 (2012)
-
Volume 60 (2011)
-
Volume 59 (2010)
-
Volume 58 (2009)
-
Volume 57 (2008)
-
Volume 56 (2007)
-
Volume 55 (2006)
-
Volume 54 (2005)
-
Volume 53 (2004)
-
Volume 52 (2003)
-
Volume 51 (2002)
-
Volume 50 (2001)
-
Volume 49 (2000)
-
Volume 48 (1999)
-
Volume 47 (1998)
-
Volume 46 (1997)
-
Volume 45 (1996)
-
Volume 44 (1996)
-
Volume 43 (1995)
-
Volume 42 (1995)
-
Volume 41 (1994)
-
Volume 40 (1994)
-
Volume 39 (1993)
-
Volume 38 (1993)
-
Volume 37 (1992)
-
Volume 36 (1992)
-
Volume 35 (1991)
-
Volume 34 (1991)
-
Volume 33 (1990)
-
Volume 32 (1990)
-
Volume 31 (1990)
-
Volume 30 (1989)
-
Volume 29 (1989)
-
Volume 28 (1989)
-
Volume 27 (1988)
-
Volume 26 (1988)
-
Volume 25 (1988)
-
Volume 24 (1987)
-
Volume 23 (1987)
-
Volume 22 (1986)
-
Volume 21 (1986)
-
Volume 20 (1985)
-
Volume 19 (1985)
-
Volume 18 (1984)
-
Volume 17 (1984)
-
Volume 16 (1983)
-
Volume 15 (1982)
-
Volume 14 (1981)
-
Volume 13 (1980)
-
Volume 12 (1979)
-
Volume 11 (1978)
-
Volume 10 (1977)
-
Volume 9 (1976)
-
Volume 8 (1975)
-
Volume 7 (1974)
-
Volume 6 (1973)
-
Volume 5 (1972)
-
Volume 4 (1971)
-
Volume 3 (1970)
-
Volume 2 (1969)
-
Volume 1 (1968)