Ralstonia spp.: emerging global opportunistic pathogens.

The bacterial genus Ralstonia (Gram-negative non-fermenters) is becoming more prevalent in cases of infection with three bacterial species, Ralstonia pickettii, Ralstonia insidiosa and Ralstonia mannitolilytica, making up all cases reported (in the literature) to date. These organisms are prevalent in many different types of water supplies (including hospital water supplies), being well adapted to survive in low-nutrient conditions. They have been shown to cause infections, sometimes serious, such as osteomyelitis and meningitis, in hospital settings. Seventy cases of R. pickettii, 13 cases of R. mannitolilytica and three cases of R. insidiosa infection have been identified from the literature. Insight is given into the types of infections that are caused by these bacteria, the underlying conditions that are associated with these infections and potential treatments.

Differentiating the growing nosocomial infectious threats Ralstonia pickettii and Ralstonia insidiosa.

Differentiation of the growing nosocomial infectious threats, Ralstonia pickettii and Ralstonia insidiosa, based on nitrate reduction, desferrioxamine susceptibility, arabinose, N-acetyl-glucosamine and phenylacetate assimilation is described. These tests can be used for preliminary identification of Ralstonia pickettii and Ralstonia insidiosa resulting in more accurate identification of these species.

Sphingomonas paucimobilis: a persistent Gram-negative nosocomial infectious organism.

Non-fermenting Gram-negative bacilli create a significant problem in clinical settings, being the most widespread cause of nosocomial infections. They are opportunistic pathogens that take advantage of underlying conditions and diseases. Sphingomonas paucimobilis, a non-fermenting Gram-negative bacillus, is regarded as of minor clinical significance; however, many instances of infections with this organism can be found in the literature. Infections include bacteraemia/septicaemia caused by contaminated solutions, e.g. distilled water, haemodialysis fluid and sterile drug solutions. Cases of pseudobacteraemia have been recorded in association with S. paucimobilis, as have many cases of unusual infections both invasive and severe, e.g. septic arthritis and osteomyelitis. No cases of death have been recorded in the literature related to S. paucimobilis. This review illustrates that S. paucimobilis is a more important pathogen than previously thought.

Development of a PCR assay for identification of the Bacillus cereus group species.

AIMS: A PCR technique was developed as a reliable and rapid identification method for the Bacillus cereus group species, based on a unique conserved sequence of the motB gene (encoding flagellar motor protein) from B. cereus, Bacillus thuringiensis and Bacillus anthracis. METHODS AND RESULTS: Primer locations were identified against eight strains of the B. cereus group spp. from nucleotide sequences available in the National Centre for Biotechnology Information database. The PCR assay was applied for the identification of 117 strains of the B. cereus group spp. and 19 strains from other microbial species, with special emphasis on foodborne pathogens. CONCLUSION: The designed cross-species primers are group specific and did not react with DNA from other Bacillus and non-Bacillus species either motile or not. The primers system enabled us to detect 10(3) CFU of B. cereus cells per millilitre of sample. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacillus cereus group spp. belongs to one of the most prevalent foodborne pathogens. Bacterial growth results in production of different toxins; therefore, consumption of food containing >10(6) bacteria per gram may result in emetic and diarrhoeal syndromes. A rapid and sensitive bacterial detection method is significant for food safety.

Ralstonia pickettii in environmental biotechnology: potential and applications.

Xenobiotic pollutants such as toluene and trichloroethylene are released into the environment by various industrial processes. Ralstonia pickettii possess significant biotechnological potential in the field of bioremediation and has demonstrated the ability to breakdown many of these toxic substances. Here, we provide a description of the major compounds that various strains of R. pickettii are capable of degrading and a brief review of their breakdown pathways and an argument for its use in bioremediation.

Ralstonia pickettii: a persistent gram-negative nosocomial infectious organism.

Non-fermenting Gram-negative bacilli create a significant problem in clinical settings, being a widespread cause of nosocomial infections. They are opportunistic pathogens that take advantage of underlying conditions and diseases. Ralstonia pickettii, a non-fermenting Gram-negative bacillus, is regarded as being of minor clinical significance; however, many instances of infections with this organism are reported in the literature. Infections can include bacteraemia/septicaemia caused by contaminated solutions, e.g. distilled water, water for injection and aqueous chlorhexidine solutions. Cases of pseudobacteraemia have been recorded in association with R. pickettii, as have many cases of unusual infections, some of which were very invasive and severe, e.g. meningitis, septic arthritis and osteomyelitis. Six cases of death in four separate instances have also been recorded related to R. pickettii. This review illustrates that R. pickettii is a more important pathogen than was thought previously.

Comparison of bioMérieux API 20NE and Remel RapID NF Plus, identification systems of type strains of Ralstonia pickettii.

UNLABELLED: A : Comparison of two commercial miniaturized rapid systems for the identification of Ralstonia pickettii strains. METHODS AND RESULTS: Varying identification results were encountered using the bioMérieux API NE system and the Remel IDS RapID NF Plus commercial systems for R. pickettii. To compare these two systems, eight strains of R. pickettii were purchased from different commercial culture collections. Additionally, 32 industrial and eight clinical isolates, initially identified using the Vitek Junior (bioMérieux) were tested. Total number of isolates tested was 48. The API 20NE identified 29 isolates, as R. pickettii but was unsuccessful with 19 isolates. The Remel IDS RapID NF Plus identified 46 isolates as R. pickettii. One clinical and one industrial isolates was identified as non-R. pickettii with both systems. CONCLUSIONS: The above results indicate that the use of API 20NE system for examining the identification of R. pickettii strains is inconsistent. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated that the RapID NF Plus is more accurate as an inexpensive identification system for the identification of R. pickettii, a potential emerging organism of medically and industrial importance.

An evaluation of five preservation techniques and conventional freezing temperatures of -20 degrees C and -85 degrees C for long-term preservation of Campylobacter jejuni.

AIMS: This study aimed to identify a simple, inexpensive preservation technique that will allow a quick and reliable recovery of Campylobacter jejuni following long-term periods of preservation. METHODS AND RESULTS: Preservation techniques include (i) Cryobank microbial preservation system using hypertonic 'cryopreservative solution' and glass beads, (ii) Cryobank microbial preservation system using defibrinated lysed horse blood and glass beads, (iii) FBP medium, (iv) 15% glycerol/85% nutrient broth no. 2 culture, and (v) 50% glycerol/50% nutrient broth no. 2 culture. Each preservation technique was evaluated over a 1-year period at conventional freezing temperatures of -20 degrees C and -85 degrees C. Replacement of 'cryopreservative fluid' in commercially prepared vials of glass beads with lysed horse blood increased the duration of preservation of Camp. jejuni by up to 6 months. CONCLUSIONS: FBP medium proved the most successful preservation technique with 100 and 80% recovery after 1 year at -85 degrees C and -20 degrees C, respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated a simple inexpensive preservation method for long-term storage of Camp. jejuni.

Comparison and evaluation of antimicrobial susceptibility testing of enterococci performed in accordance with six national committee standardized disk diffusion procedures.

Studies were conducted to compare and evaluate antimicrobial susceptibility test results for enterococci obtained by six national committee disk diffusion procedures. Variations in the incidence of isolates in resistance categories and errors were associated with the use of ciprofloxacin, gentamicin, nitrofurantoin, rifampin, and teicoplanin in a number of committee procedures. Results indicate that laboratories performing disk diffusion antimicrobial susceptibility testing may have problems correctly identifying resistance in enterococci with agents used to combat infections and that it may be difficult to compare resistance data for surveillance purposes.

Irish public perceptions and attitudes to modern biotechnology: an overview with a focus on GM foods.

This article summarizes the current situation pertaining to modern biotechnology in Ireland, with a particular focus on genetically modified (GM) crops. It briefly examines some important results of the major national surveys carried out in Ireland since 1989, highlights the recent upsurge in media (newspaper) coverage of GM related stories in three Irish opinion leader publications and it allows for an insight into the Irish public's relationship with modern biotechnology.

Expression of the gin and mom genes of bacteriophage Mu.

The gin and mom genes are located in the rightmost 1.6-kb segment, designated the beta segment, of bacteriophage Mu DNA. The gin gene is responsible for the inversion of the G segment of Mu, whereas the mom gene is involved in an unusual modification of the DNA. We have analyzed recombinant plasmids carrying one or both ends of Mu DNA for the expression of the Gin and Mom functions. The Gin protein and the presumptive Mom protein are not always detected in minicells, even though the plasmids being tested have the gin- and mom-containing segment of Mu DNA. However, some plasmids, in which the right end segment of Mu DNA is confined to the 1.6-kb beta segment, do give rise to these gene products in minicells. It seems that synthesis of the Gin and Mom proteins is inhibited in minicells, but this inhibition is lifted if most of the DNA to the left of the beta segment is eliminated from the plasmids. The most prominent Mu product detected in minicells is a 23-25-kDal polypeptide, termed here the zeta (zeta) protein. The function of the zeta protein remains unknown. In vitro transcription of Mu DNA with purified Escherichia coli RNA polymerase is limited to only two regions of the genome. The early region of Mu DNA is transcribed at a relatively high efficiency, whereas the beta region is transcribed at a low efficiency. This low-efficiency transcription appears to be specific for the gin gene; the mom gene transcript cannot be detected.

Methylation dependent expression of the mom gene of bacteriophage Mu: deletions downstream from the methylation sites affect expression.

The expression of the DNA modification gene (mom) of bacteriophage Mu requires the cellular deoxyadenosine methylase (dam) and a transactivation factor from the phage. By hypothesis, the transcription of mom is activated by methylation of three GATC sequences upstream from the mom gene. We have introduced small deletions at a fourth GATC site located about 140 base pairs downstream from the primary methylation region. Some of the deletions severely affect the mom gene expression. We propose from this analysis that (1) some important elements, possibly the promoter, concerned with the expression of mom are located between nucleotides 840 and 880 from the right end of Mu and (2) the mom protein starts with the codon GTG located at position 810. We favor the hypothesis that methylation turns off transcription upstream, thereby allowing the main mom promoter to function.