ORCID Profile
0000-0002-1153-0010
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RMIT University
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RMIT University Bundoora Campus
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Publisher: Cambridge University Press (CUP)
Date: 08-1988
DOI: 10.1017/S0950268800029265
Abstract: Two hundred and seventy-five consecutive clinical isolates of coagulase-negative staphylococci, including strains associated with disease, contaminants and skin colonizers, were speciated, tested for slime production and for their sensitivity to a range of antibiotics. Staphylococcus epidermidis was the most commonly identified species, comprising 63% of all isolates. Slime production was detected in half the strains of Staph. epidermidis, Staph. haemolyticus and Staph. Saprophyticus but was rare in other species. Most Staph. haemolyticus strains and approximately half of the Staph. epidermidis strains were resistant to five or more antibiotics. A significant association was found between slime production and multiple antibiotic resistance. For catheter-associated strains, clinical relevance was predictable by species i.e. Staph. epidermidis . Multi-resistant slime-positive Staph. haemolyticus strains, although infrequently associated with disease, were common skin colonizers, presumably acquired from the hospital environment. We describe a practical and inexpensive scheme for thespeciation of human coagulase-negative staphylococcal isolates.
Publisher: Springer Science and Business Media LLC
Date: 30-10-2010
DOI: 10.1007/S00284-010-9788-X
Abstract: Coagulase-negative staphylococci (CoNS) are the most common cause of biofilm-associated sepsis in very low birth weight infants (VLBW). Standard biofilm assays may not predict the pathogenic potential of CoNS since biofilm production is regulated by erse environmental stimuli. Staphylococcus epidermidis isolated from blood cultures from VLBW infants were evaluated for biofilm production in response to various environmental stimuli, including intravenous solutions and skin preparations. While responses to environmental stimuli were variable for in idual isolates and products, some trends were observed. Biofilm production by hospital S. epidermidis isolates (predominantly ica and biofilm-positive) was most commonly increased at 30°C and decreased in the presence of intravenous solutions and moisturisers. Commensals (mainly biofilm-negative and lacking the ica gene) were more often induced to produce biofilm than hospital isolates. These results indicate that biofilm production in S. epidermidis can vary in response to environmental stimuli encountered in the clinical setting, that standard biofilm assays are unlikely to predict clinical outcome, and that harmless skin commensals may be induced to produce biofilm by some of the products used in neonatal units.
Publisher: Oxford University Press (OUP)
Date: 29-04-2010
DOI: 10.1093/JAC/DKQ119
Abstract: (i) To evaluate the role of the adherent growth mode and extracellular polymer substance build-up in biofilm resistance to antibiotics. (ii) To re-assess various mechanisms leading to biofilm resistance to antibiotics. We compared the biofilm MICs, biofilm MBCs using the viable count method, biofilm MBCs based on broth recovery methods and minimum biofilm eradication concentrations (MBECs) of antistaphylococcal antibiotics for multilayer biofilms formed by 'biofilm-positive' S. epidermidis strains and monolayer biofilms formed by their 'biofilm-negative' mutants/variants. Bacterial densities and the quantity of persister cells in both multilayer and monolayer biofilms were assessed to evaluate their roles in biofilm resistance. Monolayer and multilayer biofilms presented similar susceptibilities to multiple antibiotics, based on biofilm MIC, broth recovery-based biofilm MBC and MBEC results. Multilayer biofilms demonstrated higher viable count-based MBCs than monolayer biofilms. Both monolayer and multilayer biofilms had very high bacterial densities of approximately 10(11-12) cfu/mL. Persister cells were found in both monolayer and multilayer biofilms, but not in planktonic cultures at log phase. The presence of persister cells in monolayer and multilayer biofilms appeared to be strain and antibiotic dependent. The adherent growth mode, rather than the ability to build up a typical multilayer biofilm structure, contributes to the high resistance of biofilms to antibiotics, and therefore might be the main virulence factor of coagulase-negative staphylococci (CoNS) with respect to antibiotic resistance. The presence of persister cells in CoNS biofilms plays an important role in antibiotic resistance. Growth at high bacterial densities is another significant factor in biofilm resistance.
Publisher: Elsevier
Date: 2001
Publisher: Microbiology Society
Date: 03-2016
DOI: 10.1099/JMM.0.000223
Publisher: Elsevier BV
Date: 05-2002
Publisher: Elsevier BV
Date: 05-2018
Publisher: Springer Science and Business Media LLC
Date: 21-12-2015
DOI: 10.1038/SREP18578
Abstract: Biofilm formation is a major pathogenicity strategy of Staphylococcus epidermidis causing various medical-device infections. Persister cells have been implicated in treatment failure of such infections. We sought to profile bacterial subpopulations residing in S. epidermidis biofilms and to establish persister-targeting treatment strategies to eradicate biofilms. Population analysis was performed by challenging single biofilm cells with antibiotics at increasing concentrations ranging from planktonic minimum bactericidal concentrations (MBCs) to biofilm MBCs (MBC biofilm ). Two populations of “persister cells” were observed: bacteria that survived antibiotics at MBC biofilm for 24/48 hours were referred to as dormant cells those selected with antibiotics at 8 X MICs for 3 hours (excluding dormant cells) were defined as tolerant-but-killable (TBK) cells. Antibiotic regimens targeting dormant cells were tested in vitro for their efficacies in eradicating persister cells and intact biofilms. This study confirmed that there are at least three subpopulations within a S. epidermidis biofilm: normal cells, dormant cells and TBK cells. Biofilms comprise more TBK cells and dormant cells than their log-planktonic counterparts. Using antibiotic regimens targeting dormant cells, i.e . effective antibiotics at MBC biofilm for an extended period, might eradicate S. epidermidis biofilms. Potential uses for this strategy are in antibiotic lock techniques and inhaled aerosolized antibiotics.
Publisher: Elsevier BV
Date: 20-12-2005
DOI: 10.1016/J.VETMIC.2005.10.012
Abstract: Traditional microbiological methods for identification of Streptococcus spp. causing bovine mastitis have been demonstrated to be less than highly reliable. PCR-RFLP analysis of 16S-23S ribosomal DNA was used to characterise seven reference strains of streptococcal mastitis pathogens as well as four reference strains of other gram-positive, catalase-negative cocci of bovine origin to allow comparative identification of field isolates. RFLP analysis of PCR products, using a combination of two restriction endonucleases in single reactions (HaeIII and AluI, HaeIII and RsaI or AluI and RsaI) generated unique patterns for species of Streptococcus, Enterococcus and Lactococcus. One hundred field isolates of Streptococcus spp. collected from cows with clinical or subclinical mastitis were tested. Fifty-seven isolates, classified by conventional tests as S. uberis, were identified as 47 S. uberis and six S. parauberis by their unique RFLP patterns. The remaining four isolates had RFLP patterns distinct from the reference strains and two of these were identified as closely related to S. iniae and two to Aerococcus viridans by 16S rRNA sequencing. Conventional identification of 17 S. agalactiae and 26 S. dysgalactiae subsp. dysgalactiae was confirmed by RFLP. Use of a combination of restriction enzymes in a single tube enabled the rapid, accurate, cost effective and easily performed identification of all major streptococcal mastitis pathogens.
Publisher: Elsevier BV
Date: 08-2010
DOI: 10.1016/J.BMCL.2010.06.020
Abstract: A series of novel polyacetylene substituted 2-hydroxy acids and derivatives were prepared and characterized. Alkylation of butane-2,3-diacetal (BDA) protected glycolic acid with iodoalkyl substituted polyacetylene compounds gave the corresponding diacetal protected polyacetylene substituted 2-hydroxy acids. Diacetal deprotection through acid mediated hydrolysis, transesterification or aminolysis afforded the 2-hydroxy-polyacetylenic acid, ester or amide derivatives. Twenty one of these novel compounds were tested against 10 microbes of clinical importance and several of them showed good antimicrobial activity, in particular against Pseudomonas aeruginosa.
Publisher: Cambridge University Press (CUP)
Date: 12-1992
DOI: 10.1017/S095026880005041X
Abstract: This study examines a series of phenotypic variants of Staphylococcus epidermidis that were generated from a pair of parent variants, isolated from valvular tissue of a patient with prosthetic valve endocarditis. The variants were initially classified by examining their colonial morphology on Congo red agar. In addition to differences in Congo red binding and colonial morphology, they differed in the expression of several surface components and enzymes. Despite these phenotypic differences, all variants had the same restriction endonuclease profile of plasmid DNA. Examination of a collection of clinical isolates demonstrated that phenotypic variation is a common property of S. epidermidis . The ability to express different combinations of surface components and enzymes could contribute to the virulence of S. epidermidis strains by enabling these organisms to colonize a range of erse environments.
Publisher: Springer Science and Business Media LLC
Date: 21-03-2016
DOI: 10.1007/S00438-016-1197-9
Abstract: Accurate and reproducible measurement of gene transcription requires appropriate reference genes, which are stably expressed under different experimental conditions to provide normalization. Staphylococcus capitis is a human pathogen that produces biofilm under stress, such as imposed by antimicrobial agents. In this study, a set of five commonly used staphylococcal reference genes (gyrB, sodA, recA, tuf and rpoB) were systematically evaluated in two clinical isolates of Staphylococcus capitis (S. capitis subspecies urealyticus and capitis, respectively) under erythromycin stress in mid-log and stationary phases. Two public software programs (geNorm and NormFinder) and two manual calculation methods, reference residue normalization (RRN) and relative quantitative (RQ), were applied. The potential reference genes selected by the four algorithms were further validated by comparing the expression of a well-studied biofilm gene (icaA) with phenotypic biofilm formation in S. capitis under four different experimental conditions. The four methods differed considerably in their ability to predict the most suitable reference gene or gene combination for comparing icaA expression under different conditions. Under the conditions used here, the RQ method provided better selection of reference genes than the other three algorithms however, this finding needs to be confirmed with a larger number of isolates. This study reinforces the need to assess the stability of reference genes for analysis of target gene expression under different conditions and the use of more than one algorithm in such studies. Although this work was conducted using a specific human pathogen, it emphasizes the importance of selecting suitable reference genes for accurate normalization of gene expression more generally.
Publisher: American Dairy Science Association
Date: 10-2010
Abstract: Cows (n=2,053) from 6 seasonally calving dairy herds were enrolled in a trial to compare the efficacy of 2 dry cow treatments. Cows received either a combination dry cow therapy of 600 mg of cloxacillin (CL) followed by an internal teat sealant (ITS) containing 2.6 g of bismuth subnitrate in all 4 quarters immediately following their final milking for the season, or only an intramammary infusion of 600 mg of CL. All cases of clinical mastitis were recorded and cultured during the first 150 d of lactation in each herd, and cow somatic cell count (SCC) was measured between 7 and 50 d postcalving. A large difference was found between treatment groups in the rate at which cows were diagnosed with clinical mastitis over the first 21 d of lactation, after which time the rate at which cows were diagnosed with clinical mastitis was similar between treatment groups. Analysis of the relative proportions of cows with clinical mastitis was performed at both the gland and cow levels. The relative risk (RR) of clinical mastitis diagnosed within 21, 30, and 100 d of calving in a gland treated with the ITS-CL combination was, respectively, 0.30 [95% confidence interval (CI)=0.21-0.44], 0.39 (0.28-0.53), and 0.58 (0.46-0.75) that of the CL group. An interaction between treatment and previous SCC was found when clinical mastitis was analyzed at the cow level. In a subset of cows that had low SCC in their previous lactation, the RR of mastitis in cows with the ITS-CL combination within 21, 30, and 100 d of calving was, respectively, 0.54 (95% CI=0.33-0.87), 0.57 (0.37-0.88), and 0.69 (0.50-0.99) that of cows that received only CL at drying off. In the subset of cows that had at least 1 high SCC in the previous lactation, the RR of mastitis in the ITS-CL combination group within 21, 30, and 100 d of calving was, respectively, 0.26 (95% CI=0.16-0.44), 0.37 (0.24-0.57), and 0.72 (0.55-0.96) that of the CL-only group. The ITS-CL combination of dry cow treatments was associated with a reduction in subclinical mastitis [SCC ≥250,000 cells/mL RR=0.80 (95% CI=0.65-0.98)] when compared with treatment with CL alone. The use of an ITS in combination with CL dry cow treatment was associated with significantly lower clinical and subclinical mastitis in the following lactation, with a greater difference found in cows that had a history of subclinical mastitis in the previous lactation.
Publisher: Microbiology Society
Date: 04-2009
Abstract: Coagulase-negative staphylococci (CoNS) are the main causative agents of bacteraemia in infants managed in neonatal intensive care units (NICUs). Intraluminal colonization of long-term central venous catheters by these bacteria and subsequent biofilm formation are the prerequisites of the bloodstream infections acquired in NICUs. The catheter lock technique has been used to treat catheter colonization however, the optimum choice of antimicrobial agents and their corresponding concentrations and exposure times have not been determined. The effectiveness of catheter lock solutions (CLSs) was assessed by determining the minimal biofilm eradication concentration of antimicrobial agents against CoNS biofilms. Five conventional antibiotics (oxacillin, gentamicin, vancomycin, ciprofloxacin and rif icin) alone or in combination, as well as ethanol, were evaluated. Ethanol was found to be superior to all of these conventional antibiotics when used as a CLS. A time–kill study and confocal laser scanning microscopy revealed that exposure to 40 % ethanol for 1 h was sufficient to kill CoNS biofilm cells. To our knowledge, this is the first in vitro study to provide solid evidence to support the rationale of using ethanol at low concentrations for a short time as a CLS, instead of using conventional antibiotics at high concentrations for a long period to treat catheter-related bloodstream infections.
Publisher: Wiley
Date: 05-2009
DOI: 10.1002/APP.30327
Publisher: American Dairy Science Association
Date: 2014
Abstract: This study was undertaken because clinicians and farmers have observed that a considerable number of cows diagnosed with Streptococcus uberis mastitis have recurrences of mastitis in the same or a different quarter. The study was an attempt to answer whether these recurring cases were due to treatment failure (in which case a search would have begun for a better treatment for Strep. uberis mastitis) or due to reinfection with a different strain of Strep. uberis. Using pulsed-field gel electrophoresis (PFGE), we determined that the majority of recurrences (20 of 27) were caused by a new strain of Strep. uberis, indicating that treatment of the initial infection had been successful. A small number of recurrences (5 of 27) were caused by the initial strain, indicating persistence. The remaining 2 recurrences occurred in a new quarter but with the initial strain of Strep. uberis, indicating either spread between quarters or reactivation of a previous subclinical infection. Analysis of the PFGE profiles failed to reveal any strain-specific propensity to persist, because strains causing recurrences occurred in most of the major clusters.
Publisher: Cambridge University Press (CUP)
Date: 10-1999
DOI: 10.1017/S0950268899002745
Abstract: Pulsed-field gel electrophoresis (PFGE) was used to investigate the epidemiology of streptococcal mastitis in dairy cattle. The most prevalent streptococcal species, Streptococcus uberis (60–80% of streptococcal isolates), was highly heterogeneous, with different cows only rarely sharing the same pulsotype. S. agalactiae was rarely encountered, however all eight isolates from one farm generated identical PFGE profiles, which differed from those of all other isolates examined, confirming cow-to-cow transmission. Fifty-two isolates of S. dysgalactiae from 27 cows on 5 farms generated 6 different profiles. However, on in idual farms, only one or two pulsotypes usually predominated. This species is generally regarded as an environmental pathogen but our data suggest that cow-to-cow transmission of S. dysgalactiae may occur. In spite of the variation in PFGE profiles of isolates from different cows, persistent infections in in idual cows were usually caused by the same pulsotype of S. uberis or S. dysgalactiae .
Publisher: American Society for Microbiology
Date: 2012
DOI: 10.1128/JCM.05124-11
Abstract: Coagulase-negative staphylococci have been identified as major causes of late-onset neonatal bacteremia in neonatal intensive care units. Sixty isolates of Staphylococcus capitis obtained from blood cultures of neonates between 2000 and 2005 were examined in this study. Biochemical analysis confirmed that 52 of these isolates belonged to the subsp. urealyticus , and the remaining 8 belonged to the subsp. capitis. Isolates of the predominant subsp. urealyticus clones were characterized by their resistance to penicillin, erythromycin, and oxacillin and their biofilm formation ability, whereas subsp. capitis isolates were generally antibiotic susceptible and biofilm negative. Pulsed-field gel electrophoresis (PFGE) after SacII digestion separated the 60 isolates into five major clusters. Sequence analysis showed that, in S. capitis , the ica operon plus the negative regulator icaR was 4,160 bp in length. PCRs demonstrated the presence of the ica operon in all isolates. Further analysis of five isolates (two biofilm-positive subsp. urealyticus , one biofilm-negative subsp. urealyticus , and two biofilm-negative subsp. capitis ) revealed that the ica operons were identical in all of the biofilm-positive subsp. urealyticus strains however, the biofilm-negative isolates showed variations. The distinctive phenotypic and genotypic characteristics revealed by this study may affect the epidemiology of the two subspecies of S. capitis in the clinical setting. These results may provide a better understanding of the contribution of these two species to bloodstream infections in neonates.
Publisher: Cambridge University Press (CUP)
Date: 10-1996
DOI: 10.1017/S0950268800001448
Abstract: The ability to produce large quantities of bioflim on solid surfaces in vitro is believed to distinguish potentially pathogenic strains of Staphylococcus epidermidis from commensals. Bioflim consists of staphylococcal cells encased in a matrix of extracellular polysaccharide (also referred to as slime), firmly adherent to each other and to the underlying surface structure. The association of slime with colonization of catheter surfaces in vivo has been examined extensively. Less attention has been paid to the contribution of slime to infections that occur in the absence of an inserted device. In a mouse model of subcutaneous infection without an implanted device 10 S. epidermidis strains (5 slime-positive, 5 slime-negative) produced abscesses thus a foreign body is not essential for the expression of virulence by S. epidermidis . Biofilm-positive strains produced significantly more abscesses, that persisted longer than biofilm-negative strains. In these chronic infections, large numbers of staphylococci were associated with macrophages and viable staphylococci were cultured from specimens of pus collected at autopsy. Thus slime or components of slime appear to delay the clearance of S. epidermidis from host tissues, possibly by interfering with intracellular killing mechanisms. However, differences in the capacity to produce abscesses, within both the slime-positive and slime-negative groups, indicate that other factors also contribute to the virulence of S. epidermidis .
Publisher: Elsevier BV
Date: 02-2015
DOI: 10.1016/J.MIMET.2014.11.012
Abstract: Clinical staphylococcus isolates possess a stronger restriction-modification (RM) barrier than laboratory strains. Clinical isolates are therefore more resistant to acceptance of foreign genetic material than laboratory strains, as their restriction systems more readily recognize and destroy foreign DNA. This stronger barrier consequently restricts genetic studies to a small number of domestic strains that are capable of accepting foreign DNA. In this study, an isolate of Staphylococcus capitis, obtained from the blood of a very low birth-weight baby, was transformed with a shuttle vector, pBT2. Optimal conditions for electro-transformation were as follows: cells were harvested at mid-log phase, electro-competent cells were prepared cells were pre-treated at 55°C for 1min 3μg of plasmid DNA was mixed with 70-80μL of competent cells (3-4×10(10)cells/mL) at 20°C in 0.5M sucrose, 10% glycerol and electroporation was conducted using 2.1kV/cm field strength with a 0.1cm gap. Compared to the conventional method, which involves DNA electroporation of Staphylococcus aureus RN4220 as an intermediate strain to overcome the restriction barrier, our proposed approach exhibits a higher level (3 log10 units) of transformation efficiency. Heat treatment was used to temporarily inactivate the recipient RM barrier. Other important parameters contributing to improved electro-transformation efficiency were growth stage for cell harvesting, the quantity of DNA, the transformation temperature and field strength. The approach described here may facilitate genetic manipulations of this opportunistic pathogen.
Publisher: Elsevier BV
Date: 2016
DOI: 10.1016/J.CMI.2015.09.008
Abstract: Nosocomial late-onset sepsis represents a frequent cause of morbidity and mortality in preterm neonates. The Staphylococcus capitis clone NRCS-A has been previously described as an emerging cause of nosocomial bacteraemia in French neonatal intensive-care units (NICUs). In this study, we aimed to explore the possible unrecognized dissemination of this clone on a larger geographical scale. One hundred methicillin-resistant S. capitis strains isolated from neonates (n = 86) and adult patients (n = 14) between 2000 and 2013 in four different countries (France, Belgium, the UK, and Australia) were analysed with SmaI pulsed-field gel electrophoresis (PFGE) and dru typing. The vast majority of NICU strains showed the NRCS-A pulsotype and the dt11c type (96%). We then randomly selected 14 isolates (from neonates, n = 12, three per country from adult patients, n = 2), considered to be a subset of representative isolates, and performed further molecular typing (SacII PFGE, SCCmec typing, and multilocus sequence typing-like analysis), confirming the clonality of the S. capitis strains isolated from neonates, despite their distant geographical origin. Whole genome single-nucleotide polymorphism-based phylogenetic analysis of five NICU isolates (from the different countries) attested to high genetic relatedness within the NRCS-A clone. Finally, all of the NRCS-A strains showed multidrug resistance (e.g. methicillin and aminoglycoside resistance, and decreased vancomycin susceptibility), with potential therapeutic implications for infected neonates. In conclusion, this study represents the first report of clonal dissemination of methicillin-resistant coagulase-negative Staphylococcus clone on a large geographical scale. Questions remain regarding the origin and means of international spread, and the reasons for this clone's apparent predilection for neonates.
Publisher: Elsevier BV
Date: 04-2001
DOI: 10.1046/J.1469-0691.2001.00214.X
Abstract: To analyze Staphylococcus epidermidis strains, previously tested for their virulence in a mouse model of subcutaneous infection, for various phenotypic traits (biofilm density, extracellular polysaccharide, slime-associated antigen (SAA)) and for the presence of the ica gene cluster, to determine which of these phenotypic and genotypic methods best correlates with virulence in the mouse model. The quantitative biofilm assay was performed on 10 strains of S. epidermidis, comprising (1) RP62A (ATCC 35984), (2) the strongest and weakest biofilm producers in our collection, (3) a pair of phenotypic variants, and (4) a strain whose biofilm density was enhanced in iron-limited media. Biofilm density was measured after growth at 37 degrees C and at ambient temperature, in trypticase soy broth (TSB) with and without glucose supplementation and using both chemical and heat fixation. Strains were assayed for SAA using a double immunodiffusion method. Extracellular polysaccharide was detected by transmission electron microscopy (TEM). A 546-base-pair segment of the ica gene cluster was lified by PCR. Biofilm formation in TSB, glucose-enriched TSB, extracellular polysaccharide (observed by TEM), expression of SAA and presence of the ica gene predicted virulence of nine, nine, nine, eight and eight of 10 strains, respectively. The phenotypic expression of biofilm and related properties was medium and temperature dependent. We encountered one ica-positive strain that failed to express biofilm in standard TSB at 37 degrees C, but was virulent in a mouse model, and another strain that lacked ica, produced biofilm and was virulent in the model. Mouse virulence in our model can be predicted by any of the phenotypic or genotypic methods examined for > or = 80% of strains. Medium and incubation conditions affect the expression of phenotypic markers by some strains. For the remaining strains, possible reasons for inconsistencies between the presence of the ica gene, phenotypic markers and mouse virulence include (1) dependence of biofilm on genes other than ica, (2) sequence differences in ica, (3) dependence of biofilm expression in vivo on strain characteristics and media used to prepare inocula for in vivo studies.
Publisher: Informa UK Limited
Date: 07-2013
DOI: 10.1080/08927014.2013.800192
Abstract: Biofouling, the unwanted growth of sessile microorganisms on submerged surfaces, presents a serious problem for underwater structures. While biofouling can be controlled to various degrees with different microstructure-based patterned surfaces, understanding of the underlying mechanism is still imprecise. Researchers have long speculated that microtopographies might influence near-surface microfluidic conditions, thus microhydrodynamically preventing the settlement of microorganisms. It is therefore very important to identify the microfluidic environment developed on patterned surfaces and its relation with the antifouling behaviour of those surfaces. This study considered the wall shear stress distribution pattern as a significant aspect of this microfluidic environment. In this study, patterned surfaces with microwell arrays were assessed experimentally with a real-time biofilm development monitoring system using a novel microchannel-based flow cell reactor. Finally, computational fluid dynamics simulations were carried out to show how the microfluidic conditions were affecting the initial settlement of microorganisms.
Publisher: Springer Science and Business Media LLC
Date: 27-05-2010
Abstract: Coagulase-negative staphylococci are major causes of bloodstream infections in very low birth weight babies cared for in Neonatal Intensive Care Units. The virulence of these bacteria is mainly due to their ability to form biofilms on indwelling medical devices. Biofilm-related infections often fail to respond to antibiotic chemotherapy guided by conventional antibiotic susceptibility tests. Coagulase-negative staphylococcal blood culture isolates were grown in different phases relevant to biofilm formation: planktonic cells at mid-log phase, planktonic cells at stationary phase, adherent monolayers and mature biofilms and their susceptibilities to conventional antibiotics were assessed. The effects of oxacillin, gentamicin, and vancomycin on preformed biofilms, at the highest achievable serum concentrations were examined. Epifluorescence microscopy and confocal laser scanning microscopy in combination with bacterial viability staining and polysaccharide staining were used to confirm the stimulatory effects of antibiotics on biofilms. Most coagulase-negative staphylococcal clinical isolates were resistant to penicillin G (100%), gentamicin (83.3%) and oxacillin (91.7%) and susceptible to vancomycin (100%), ciprofloxacin (100%), and rif icin (79.2%). Bacteria grown as adherent monolayers showed similar susceptibilities to their planktonic counterparts at mid-log phase. Isolates in a biofilm growth mode were more resistant to antibiotics than both planktonic cultures at mid-log phase and adherent monolayers however they were equally resistant or less resistant than planktonic cells at stationary phase. Moreover, for some cell-wall active antibiotics, concentrations higher than conventional MICs were required to prevent the establishment of planktonic cultures from biofilms. Finally, the biofilm-growth of two S. capitis isolates could be enhanced by oxacillin at the highest achievable serum concentration. We conclude that the resistance of coagulase-negative staphylococci to multiple antibiotics initially remain similar when the bacteria shift from a planktonic growth mode into an early attached mode, then increase significantly as the adherent mode further develops. Furthermore, preformed biofilms of some CoNS are enhanced by oxacillin in a dose-dependent manner.
Publisher: S. Karger AG
Date: 2008
DOI: 10.1159/000171361
Abstract: The human gut flora is a complex and finely balanced ecosystem which plays an important protective role in humans. Although relatively stable, its composition may be altered in various disease states and by the administration of antimicrobial agents. Preparations containing viable lactic acid bacteria of human origin appear to have value in restoring normal microbial function and alleviating symptoms in some patients with gastrointestinal infection and other conditions.
Publisher: American Chemical Society (ACS)
Date: 16-04-2018
Publisher: IWA Publishing
Date: 18-04-2011
DOI: 10.2166/WH.2011.134
Abstract: To assess microbial safety of treated sewage sludge (biosolids), we examined the inactivation of microbial indicators for potential bacterial, viral and protozoan pathogens. The levels of indicators were determined throughout the air-drying and storage phases of anaerobically digested sewage sludge. S les were collected from two wastewater treatment plants (WWTPs) in Victoria, Australia. Established methods were applied for analysis of bacteria and coliphages, based on membrane filtration and layered plates, respectively. In the pan drying phase, the prevalence of Escherichia coli was reduced by & log10 compared with sludge entering the pan. Thus, after pan drying of 8-11 months at WWTP A and 15 months at WWTP B, the numbers of E. coli were reduced to below 102 cfu/g dry solids (DS). This level is acceptable for unrestricted use in agriculture in Australia (P1 treatment grade), the UK (enhanced treatment status) and the USA (Class A pathogen reduction). Coliphage numbers also decreased substantially during the air-drying phase, indicating that enteric viruses are also likely to be destroyed during this phase. Clostridium perfringens appeared to be an overly conservative indicator. Survival, but not regrowth, of E. coli or Salmonella was observed in rewetted biosolids (15–20% moisture content), after being seeded with these species, indicating a degree of safety of stored biosolids upon rewetting by rain.
Publisher: American Society for Microbiology
Date: 27-08-2015
Abstract: Staphylococcus capitis pulsotype NRCS-A was previously reported as a frequent cause of late-onset sepsis in neonatal intensive care units (NICUs) worldwide. Here, we report the whole-genome shotgun sequences of four S. capitis pulsotype NCRS-A strains, CR03, CR04, CR05, and CR09, isolated from Belgium, Australia, the United Kingdom, and France, respectively.
Publisher: Microbiology Society
Date: 06-2015
DOI: 10.1099/JMM.0.000059
Abstract: The ica operon encoding polysaccharide intercellular adhesion, which facilitates biofilm formation in staphylococci, has been extensively studied in Staphylococcus epidermidis and Staphylococcus aureus. Based on in silico analysis, we suggest the following functional model for Ica proteins in S. capitis. IcaA is responsible for polysaccharide synthesis. IcaA and IcaD complete transferring the growing sugar chain to the cell surface IcaB is a deacetylase, with the same function as IcaB of S. epidermidis. IcaC mainly modifies the synthesized glucan by acetylation. We also examined the effects of subinhibitory concentrations of erythromycin on phenotypic biofilm expression and transcription of biofilm-related genes, using isolates representing the two subspecies of Staphylococcus capitis and different biofilm and resistance phenotypes. On induction with erythromycin, biofilm density was strongly elevated in two erythromycin-resistant S. capitis, but not in three susceptible isolates. In the representative erythromycin-resistant S. capitis subsp. urealyticus, there were significant upregulations of the icaA gene and its positive regulator sarA on transition to the stationary phase without erythromycin induction. There were also significant increases in the transcription levels of icaA, rsbU and sigB corresponding to a very strong biofilm phenotype in the stationary phase on erythromycin stress. In contrast, the representative erythromycin-susceptible S. capitis subsp. capitis displayed upregulation only of altE on entry into the stationary phase with erythromycin induction, but this change was not associated with enhancement of biofilm production. These findings suggest that the two subspecies of S. capitis adopt different pathogenesis and survival strategies to adapt to a hostile environment.
Publisher: Elsevier BV
Date: 06-2018
Publisher: Elsevier BV
Date: 05-1999
DOI: 10.1016/S0378-1135(99)00020-6
Abstract: Mastitis caused by Staphylococcus aureus is a disease of major economic importance to the dairy industry. Transmission occurs during milking, with chronically-infected cows acting as the major reservoirs of infection. PCR-coagulase gene typing of 151 S. aureus isolates from seven farms generated only six PCR types, with 110 (73%) isolates assigned to PCR type 1 and 23 (15.2%) isolates assigned to type 2. PCR type 1 was the predominant type on five of the seven farms, including farms in geographically separated regions of Victoria, while type 2 predominated on two farms. With the exception of the 41 isolates from one farm, all isolates were resistant to penicillin, but susceptible to other antibiotics that are routinely used to manage mastitis in dairy cattle. Nine of 11 cows with chronic S. aureus infection showed evidence of persistence of a single PCR type for periods of up to 9 months. Two different PCR types of S. aureus were isolated from the other two cows over the same period.
Publisher: Elsevier BV
Date: 2018
Publisher: American Society for Microbiology
Date: 2008
DOI: 10.1128/AEM.01373-07
Abstract: Multilocus sequence typing analysis of Streptococcus uberis has identified a cluster of isolates associated with clinical and subclinical mastitis and a cluster associated with cows with low somatic cell counts in their milk. Specific groups of genotypes (global clonal complex [GCC] sequence type 5s [ST5s] and GCC ST143s) were highly associated ( P = 0.006) with clinical and subclinical mastitis and may represent a lineage of virulent isolates, whereas isolates belonging to GCC ST86 were associated with low-cell-count cows. This study has, for the first time, demonstrated the occurrence of identical sequence types (ST60 and ST184) between different continents (Australasia and Europe) and different countries (Australia and New Zealand). The standardized index of association and the empirical estimation of the rate of recombination showed substantial recombination within the S. uberis population in Australia, consistent with previous multilocus sequence type analyses.
Publisher: Frontiers Media SA
Date: 17-05-2016
Publisher: Springer Science and Business Media LLC
Date: 06-04-2006
DOI: 10.1007/S10096-006-0130-2
Abstract: Selected coagulase-negative staphylococci from the blood of very-low-birth-weight infants in the Neonatal Intensive Care Unit at the Royal Women's Hospital, Melbourne, collected over a 5-year period were examined. Isolates were classified as invasive or contaminants, speciated, typed by pulsed-field gel electrophoresis, and examined for biofilm genes (icaA, icaC, and icaD), adhesion genes (atlE, fbe), and the number of copies of IS256. Of the 24 isolates studied, there were 13 contaminants and 11 invasive isolates. The collection included 15 Staphylococcus epidermidis, eight Staphylococcus capitis, and one each of Staphylococcus warneri and Staphylococcus haemolyticus. Two small clusters of S. epidermidis that belonged to the same molecular type were identified. All S. capitis isolates belonged to the same molecular type or subtype, suggesting that a particular clone was circulating in the unit. There was no significant difference in the species found, the presence of icaA, icaC, icaD, atlE, or fbe, or the number of copies of IS256 between invasive isolates and contaminants. A series of nasal isolates from nonhospitalized adults differed from hospital isolates in the absence of IS256 and the low prevalence of icaC. There was no evidence of IS256-mediated insertion into ica genes as a mechanism of phase variation. These findings suggest that contaminants and invasive isolates derived from the same pool of hospital strains capable of causing sepsis in compromised hosts and that other mechanisms of phase variation exist, apart from IS256 insertion into ica genes.
Publisher: Springer London
Date: 1990
Publisher: American Society for Microbiology
Date: 09-2008
DOI: 10.1128/JCM.00592-08
Abstract: Nine Staphylococcus capitis isolates from blood cultures of newborns were examined for resistance to vancomycin. MICs were within the susceptible range, but population profiling revealed a resistant subpopulation. Only isolates with the largest subpopulation were identified as heteroresistant to vancomycin by Etest. This finding may have therapeutic implications.
No related grants have been discovered for Margaret Deighton.