Biomedical subjects

Warren B Grubb

Publications and source records attributed to Warren B Grubb.

7 recordsLinked to original sources

PubMed2003

Characterization of drug-resistant Staphylococcus aureus isolated from poultry processing plants in Western Australia.

Poultry processing plants can provide a favourable environment for the survival and transmission of Staphylococcus aureus. It is known that infections due to antibiotic-resistant strains of S. aureus are an increasingly serious problem clinically and, since antibiotic exposure in food-animal species may lead to antibiotic-resistant bacteria, it is possible that processed poultry may constitute a reservoir for disseminating antibiotic-resistance into the community. The present study was undertaken to determine the prevalence of antimicrobial-resistant S. aureus in two poultry processing plants, and to characterize the isolates by antimicrobial susceptibility and chromosomal and plasmid DNA analysis. One hundred and twenty-six S. aureus were isolated from two poultry processing plants in Western Australia. All were sensitive to 14 of the 26 antimicrobials tested and all isolates were resistant to at least one antibiotic and one chemical marker, the prominent resistance combination being to penicillin and cadmium (89%). Forty-six (36.5%) of the isolates were resistant to six or more of the antimicrobial agents tested. Overall there were no consistent resistance patterns for the isolates and no consistent patterns were found between and within the two processing plants. There were 24 epidemiologically unrelated Sma1 contour-clamped homogeneous electric field (CHEF) groups and 17 different plasmid profiles detected among the isolates. All isolates were found to harbour from between one to seven plasmids. The majority of isolates carried at least one large plasmid (22-48 Kb), and one or more small plasmids (1-3 Kb). Some isolates with epidemiologically related CHEF patterns had similar plasmid profiles and resistance patterns.

Animals↗

PubMed2003

Alterations in phage-typing patterns in vancomycin-intermediate Staphylococcus aureus.

The ability of phage-typing and SmaI chromosomal RFLPs to conclude appropriate strain relatedness between a collection of 12 well-characterized in vitro-selected vancomycin-intermediate Staphylococcus aureus (VISA) strains and their seven vancomycin-susceptible parent strains is reported. Generally, no SmaI RFLP alterations were observed in VISA strains when they were compared with their respective parent strains, and clonal relationships between isogenic strains were clearly evident. Unlike the SmaI RFLP results, parent strains and VISA derivatives generally did not share similar phage-typing profiles. Depending on the phage set investigated, some VISA strains even became untypable by this method. Loss of phage infectivity is probably due to cell wall (phage receptor) alterations that are expressed by the VISA strains investigated. Collectively, these findings indicate that inappropriate relationships between VISA and vancomycin-susceptible parents might be drawn if only phage-typing and antibiotic susceptibility are utilized to determine epidemiological relationships.

Anti-Bacterial Agents↗

PubMed2003

Are all community methicillin-resistant Staphylococcus aureus related? A comparison of their mec regions.

AIMS: To compare the relationship of community-acquired, methicillin-resistant Staphylococcus aureus (CMRSA) from five Australian States and New Zealand. METHODS: Contour-clamped homogeneous electric field (CHEF) electrophoresis and analysis of the mec complex and ccr gene complex by PCR were used to compare 22 CMRSA isolates from Western Australia (WA), South Australia (SA), Victoria (VIC), New South Wales (NSW) and New Zealand (NZ) and three hospital-acquired epidemic MRSA (EMRSA). RESULTS: Sixteen community isolates were found to carry Class B mec complex and Type 2 ccr gene complex. Two WA isolates carried the Class B1 mec complex and three VIC and one SA isolate carried a previously unreported mec complex, which has been labelled E. The ccr gene type of the Class B1 and Class E isolates could not be determined. These isolates may carry previously unreported ccr gene complexes. The relatedness of the CHEF patterns of the CMRSA was dependent on their geographical origin. A similar CHEF pattern was found in some WA MRSA, VIC and SA isolates. NSW and NZ CMRSA had the same CHEF patterns and were similar to three VIC isolates and EMRSA-16. Two SA CMRSA isolates had CHEF patterns similar to the English EMRSA-15 strain. A multiply resistant, nosocomial EMRSA from Australia had a class A mec complex, and a CHEF pattern, which was unrelated to any of the CMRSA. CONCLUSION: Most of the CMRSA isolated from five Australian states and New Zealand had unrelated CHEF patterns. However, the majority of them carried the Type IV SCCmec cassette (Class B mec and Type 2 ccr gene complexes), which indicates that they may have acquired their mec complex from the same source or that they have evolved from the same progenitor. Some of the CMRSA had a previously undescribed SCCmec cassette.

Australia↗

PubMed2002

Proteome analysis of highly immunoreactive proteins of Helicobacter pylori.

BACKGROUND: Identification of the immunoreactive proteins of Helicobacter pylori is important for the development of both diagnostic tests and vaccines relating to the organism. Our aim was to determine whether there are significant differences between human IgG and IgA reactivities to individual H. pylori proteins, and whether patterns of immunoreactivity are sustained across different strains of H. pylori. METHOD: The total complement of protein from seven strains of H. pylori was resolved by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Proteins were transferred electrophoretically onto polyvinylene difluoride (PVDF) membranes, which were probed with sera pooled either from H. pylori-infected patients, or noninfected (control) patients. Highly immunoreactive proteins were detected using chromogenic enzyme-antibody conjugates recognising either serum IgG or IgA. These proteins were then characterised by tryptic peptide-mass fingerprinting using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). RESULTS: Highly immunoreactive proteins were detected which were common to all seven strains, and recognised by both immunoglobulin subclasses. The proteins appear to be localised in five groups. Protein analysis established that these groups encompass multiple isoforms of chaperonin HspB (two subgroups); urease beta-subunit UreB; elongation factor EF-Tu; and flagellin FlaA. The pattern of highly immunoreactive proteins was strongly conserved across the seven strains. CONCLUSION: These results suggest that within a tightly defined region on the H. pylori proteome map there are five groups of proteins that are highly reactive to both IgG and IgA. Our analysis suggests it is unlikely that the highly immunoreactive clusters harbour any significant proteins other than isoforms of HspB, UreB, EF-Tu and FlaA, and that, with the partial exception of FlaA, these clusters are strongly conserved across all seven strains.

Bacterial Proteins↗

PubMed2002

Genetic characterization of the fusidic acid and cadmium resistance determinants of Staphylococcus aureus plasmid pUB101.

We report the cloning of the fusidic acid and cadmium resistance determinants from Staphylococcus aureus plasmid pUB101. The pUB101 fusidic acid resistance determinant was located on a 2.9 kb HindIII fragment. Sequencing of this fragment revealed three putative open reading frames (ORFs) of 213 (far1), 152 (orf152) and 170 amino acids (orf170), which are flanked by the right-hand end of insertion sequence IS431/257 (IS431/257RH) and a partial ORF. Far1 and Orf152 demonstrated homology with a chromosomally encoded fibronectin-binding protein of Listeria monocytogenes and the putative protein YosT, found on the SP beta c2 prophage of Bacillus subtilis, respectively. Transformation of S. aureus with a construct containing a 949 bp far1-specific amplicon led to the isolation of a fusidic acid-resistant transformant, thereby identifying the pUB101 fusidic acid resistance structural gene. Between orf152 and far1 we identified a unique 113 bp symmetrical element and other repeat elements that may be involved with the control of orf152 and/or far1 expression. Hybridization of Southern blots revealed that far1 was not located on the chromosome or plasmid content of a limited number of Australian, UK and Hong Kong fusidic acid-resistant isolates. The pUB101 cadmium resistance determinant was located on a 3.6 kb HindIII fragment that carried a cadDX operon, remnants of two putative plasmid replication protein genes and IS431/257RH. Sequence analysis also demonstrated the presence of a single-stranded origin of replication, normally found on rolling circle replicating plasmids, within the putative promoter region of the cadDX operon.

Anti-Bacterial Agents↗

PubMed2002

Genetic organization of mecA and mecA-regulatory genes in epidemic methicillin-resistant Staphylococcus aureus from Australia and England.

The mecA gene that encodes methicillin resistance in Staphylococcus aureus may be regulated by the mecR1 and mecI genes, and this region has been referred to as the mec complex. An analysis of these regulatory genes in 35 epidemic methicillin-resistant S. aureus (MRSA) isolated in England and Australia has found that they contain three classes of mec complex. Firstly, the Class A mec complex with complete mecR1 and mecI genes. Secondly, a new variant of Class A, the Class A1 mec complex, with a 166 bp deletion in the membrane-spanning domain of mecR1 and a complete mecI gene. Thirdly, the Class B mec complex, in which the penicillin-binding domain of mecR1 and the whole mecI gene are deleted by the insertion of a partial sequence of IS1272. Seven MRSA isolated in England and Australia over different time periods had the Class A mec complex. However, the isolates did not have closely related pulsed-field gel electrophoresis (PFGE) patterns. The Class A1 mec complex was found in 12 Australian isolates and the English epidemic MRSA, EMRSA-1. All these organisms were isolated in the early 1980s and had closely related PFGE patterns. The Class B mec complex region was found in nine EMRSA and seven Australian MRSA isolated over the period from the 1970s to 2000. These isolates had related PFGE patterns. The mecA region was also compared in the isolates and all but two of the isolates had an XbaI restriction site. These results support the global spread of epidemic clones and confirm the close relationship between the Australian and English MRSA strains.

Australia↗

PubMed2002

Dissemination of new methicillin-resistant Staphylococcus aureus clones in the community.

Multiple methicillin-resistant Staphylococcus aureus (MRSA) clones carrying type IV staphylococcal cassette chromosome mec were identified in the community-acquired MRSA strains of both the United States and Australia. They multiplied much faster than health-care-associated MRSA and were resistant to fewer non-beta-lactam antibiotics. They seem to have been derived from more diverse S. aureus populations than health-care-associated MRSA strains.

Anti-Bacterial Agents↗