ORCID Profile
0000-0002-2118-1651
Current Organisation
University of Adelaide
Does something not look right? The information on this page has been harvested from data sources that may not be up to date. We continue to work with information providers to improve coverage and quality. To report an issue, use the Feedback Form.
Publisher: Public Library of Science (PLoS)
Date: 25-07-2022
DOI: 10.1371/JOURNAL.PONE.0271912
Abstract: Haemophilus influenzae , Streptococcus pneumoniae and Moraxella catarrhalis are bacterial species which frequently co-colonise the nasopharynx, but can also transit to the middle ear to cause otitis media. Chronic otitis media is often associated with a polymicrobial infection by these bacteria. However, despite being present in polymicrobial infections, the molecular interactions between these bacterial species remain poorly understood. We have previously reported competitive interactions driven by pH and growth phase between H . influenzae and S . pneumoniae . In this study, we have revealed competitive interactions between the three otopathogens, which resulted in reduction of H . influenzae viability in co-culture with S . pneumoniae and in triple-species culture. Transcriptomic analysis by mRNA sequencing identified a central role of arginine in mediating these interactions. Arginine supplementation was able to increase H . influenzae survival in a dual-species environment with S . pneumoniae , and in a triple-species environment. Arginine was used by H . influenzae for ATP production, and levels of ATP generated in dual- and triple-species co-culture at early stages of growth were significantly higher than the combined ATP levels of single-species cultures. These results indicate a central role for arginine-mediated ATP production by H . influenzae in the polymicrobial community.
Publisher: Oxford University Press (OUP)
Date: 15-07-2009
DOI: 10.1086/599987
Publisher: American Society for Microbiology
Date: 06-2006
Abstract: Neisseria gonorrhoeae is a host-adapted pathogen that colonizes primarily the human genitourinary tract. This bacterium encounters reactive oxygen and reactive nitrogen species as a consequence of localized inflammatory responses in the urethra of males and endocervix of females and also of the activity of commensal lactobacilli in the vaginal flora. This review describes recent advances in the understanding of defense systems against oxidative stress in N. gonorrhoeae and shows that while some of its defenses have similarities to the paradigm established with Escherichia coli , there are also some key differences. These differences include the presence of a defense system against superoxide based on manganese ions and a glutathione-dependent system for defense against nitric oxide which is under the control of a novel MerR-like transcriptional regulator. An understanding of the defenses against oxidative stress in N. gonorrhoeae and their regulation may provide new insights into the ways in which this bacterium survives challenges from polymorphonuclear leukocytes and urogenital epithelial cells.
Publisher: Elsevier BV
Date: 2016
DOI: 10.1016/J.RESMIC.2015.08.004
Abstract: An alcohol dehydrogenase, AdhC, is required for Haemophilus influenzae Rd KW20 growth with high oxygen. AdhC protects against both exogenous and metabolically generated, endogenous reactive aldehydes. However, adhC in the strain 86-028NP is a pseudogene. Unlike the Rd KW20 adhC mutant, 86-028NP does grow with high oxygen. This suggests the differences between Rd KW20 and 86-028NP include broader pathways, such as for the maintenance of redox and metabolism that avoids the toxicity related to oxygen. We hypothesized that these differences affect their resistance to relevant toxic chemicals, including reactive aldehydes. Across a range of oxygen concentrations, despite the growth profiles of Rd KW20 and 86-028NP being similar, there was a significant variation in their sensitivity to reactive aldehydes. 86-028NP is more sensitive to methylglyoxal, formaldehyde and glycolaldehyde under high oxygen than low oxygen as well as compared to Rd KW20. Also, as oxygen levels changed the whole genome gene expression profiles of Rd KW20 and 86-028NP revealed distinctions in their transcriptomes (the iron, FNR and ArcAB regulons). These were indicative of a difference in their intracellular redox properties and we show it is this that underpins their survival against reactive aldehydes.
Publisher: Oxford University Press (OUP)
Date: 2022
Abstract: Enterococcus faecalis is able to adapt to alkaline conditions and is commonly recovered from teeth in which endodontic treatment has failed. The role that E. faecalis membrane proteins play in survival strategies to extreme alkaline conditions is unclear. We grew E. faecalis V583 in a chemostat at pH 8 and 11 at one-tenth the organism’s relative maximum growth rate. Following membrane shaving, isotope-coding protein labels were added at the peptide level to s les and then combined. The relative proportion of membrane proteins were identified using LC-ESI mass spectrometry and MaxQuant analysis. Ratios of membrane proteins were log2 transformed, with proteins deviating by more than 1 SD of the mean considered to be up- or down-regulated. A total of six proteins were up-regulated in pH 11 including: EF0669 (polysaccharide biosynthesis family) EF1927 (glycerol uptake facilitator), and EF0114 (glycosyl hydrolase). A total of five proteins were down-regulated including: EF0108 (C4-dicarboxylate transporter) EF1838 (PTS system IIC component) EF0456 (PTS system IID component) and EF0022 (PTS mannose-specific IID component). In extreme alkaline conditions, the membrane proteins of E. faecalis seem to be involved in a shift of carbohydrate metabolism from the PTS system to glycerol, which supports the formation of a protective capsule protecting the cell.
Publisher: Royal Society of Chemistry (RSC)
Date: 2021
DOI: 10.1039/D1TB01122G
Abstract: Antibacterial activity of selected 2D materials on porous-titania prepared by plasma electrolytic oxidation (PEO) is presented.
Publisher: Elsevier
Date: 2011
Publisher: Elsevier BV
Date: 2021
Publisher: Elsevier BV
Date: 12-2015
DOI: 10.1016/J.MEEGID.2015.10.011
Abstract: A key to persistent and recurrent Staphylococcus aureus infections is its ability to adapt to erse and toxic conditions. This ability includes a switch into a biofilm or to the quasi-dormant Small Colony Variant (SCV). The development and molecular attributes of SCVs have been difficult to study due to their rapid reversion to their parental cell-type. We recently described the unique induction of a matrix-embedded and stable SCV cell-type in a clinical S. aureus strain (WCH-SK2) by growing the cells with limiting conditions for a prolonged timeframe. Here we further study their characteristics. They possessed an increased viability in the presence of antibiotics compared to their non-SCV form. Their stability implied that there had been genetic changes we therefore determined both the genome sequence of WCH-SK2 and its stable SCV form at a single base resolution, employing Single Molecular Real-Time (SMRT) sequencing that enabled the methylome to also be determined. The genetic features of WCH-SK2 have been identified the SCCmec type, the pathogenicity and genetic islands and virulence factors. The genetic changes that had occurred in the stable SCV form were identified most notably being in MgrA, a global regulator, and RsbU, a phosphoserine phosphatase within the regulatory pathway of the sigma factor SigB. There was a shift in the methylomes of the non-SCV and stable SCV forms. We have also shown a similar induction of this cell-type in other S. aureus strains and performed a genetic comparison to these and other S. aureus genomes. We additionally map RNAseq data to the WCH-SK2 genome in a transcriptomic analysis of the parental, SCV and stable SCV cells. The results from this study represent the unique identification of a suite of epigenetic, genetic and transcriptional factors that are implicated in the switch in S. aureus to its persistent SCV form.
Publisher: MDPI AG
Date: 28-06-2023
DOI: 10.3390/ANTIBIOTICS12071122
Abstract: The extent of similarity between E. faecium strains found in healthy feedlot beef cattle and those causing extraintestinal infections in humans is not yet fully understood. This study used whole-genome sequencing to analyse the antimicrobial resistance profile of E. faecium isolated from beef cattle (n = 59) at a single feedlot and compared them to previously reported Australian isolates obtained from pig (n = 60) and meat chicken caecal s les (n = 8), as well as human sepsis cases (n = 302). The E. faecium isolated from beef cattle and other food animal sources neither carried vanA/vanB responsible for vancomycin nor possessed gyrA arC and liaR/liaS gene mutations associated with high-level fluoroquinolone and daptomycin resistance, respectively. A small proportion (7.6%) of human isolates clustered with beef cattle and pig isolates, including a few isolates belonging to the same sequence types ST22 (one beef cattle, one pig, and two human isolates), ST32 (eight beef cattle and one human isolate), and ST327 (two beef cattle and one human isolate), suggesting common origins. This provides further evidence that these clonal lineages may have broader host range but are unrelated to the typical hospital-adapted human strains belonging to clonal complex 17, significant proportions of which contain vanA/vanB and liaR/liaS. Additionally, none of the human isolates belonging to these STs contained resistance genes to WHO critically important antimicrobials. The results confirm that most E. faecium isolated from beef cattle in this study do not pose a significant risk for resistance to critically important antimicrobials and are not associated with current human septic infections.
Publisher: American Society for Microbiology
Date: 18-10-2022
DOI: 10.1128/JB.00138-22
Abstract: Within a bacterial population, a stochastically generated heterogeneity of phenotypes allows continual survival against current and future stressors. The generation of a sub-population of quasi-dormant Small Colony Variants (SCV) in Staphylococcus aureus is such a mechanism, allowing for persistent or relapse of infection despite initial intervention seemingly clearing the infection.
Publisher: Elsevier BV
Date: 04-2023
Publisher: American Society for Microbiology
Date: 03-2007
DOI: 10.1128/IAI.01634-06
Abstract: The adhC gene from 11 strains of Neisseria gonorrhoeae was distinguished from its homologue in Neisseria meningitidis by the presence of a premature stop codon caused by a single base insertion. Mutational analysis showed that NADH S -nitrosoglutathione oxidoreductase activity was associated with adhC in Neisseria meningitidis but not in Neisseria gonorrhoeae .
Publisher: Oxford University Press (OUP)
Date: 10-2018
Abstract: Haemophilus influenzae and Streptococcus pneumoniae are known aetiologic agents of chronic otitis media, frequently as a multispecies infection. In this study, we show that the outcome of H. influenzae/S. pneumoniae interactions is dependent on the nutrient source. In continuous culture containing chemically defined media with lactose, S. pneumoniae was non-viable in mono-culture, and in co-culture remained non-viable until 288 h. With glucose, S. pneumoniae became non-viable in mono-culture, but uniquely existed in 3 distinct states in co-culture: parental cells (until 24 h), a dormant state until 336 h and its re-emergence as a non-mucoidal, small colony variant (SCV). The S. pneumoniae SCV was stable and whole genome sequencing showed three major single nucleotide polymorphisms in the SCV cells-cap3A (capsule biosynthesis pathway), fpg (DNA glycosylase of the DNA repair mechanism) and glutamate-5-kinase. Previously, fpg mutants have shown increased mutator rates, permitting bacterial survival against host-generated stresses. Transcriptomics showed these SCV cells up-regulated sugar transporters and toxin/antitoxin systems. An animal model revealed a reduced survival in the lungs and ear by SCV cells. This is the first study documenting the effect of carbon source and the development of a distinct S. pneumoniae cell type during H. influenzae/S. pneumoniae interactions.
Publisher: Elsevier BV
Date: 02-2020
DOI: 10.1016/J.JENVMAN.2019.109744
Abstract: Methane production via anaerobic digestion of poultry litter provides a pathway for energy production from an abundant waste product. Recent studies have shown the use of biochar (pyrolysed biomass) can decrease methane production lag times and increase peak daily yields from ammonia-stressed low-solids anaerobic digesters. Due to the variety of feedstocks and digester configurations used, research to date has not yet determined the effect of biochar addition as a function of the digester total solids content. This study shows the addition of biochar reduces the lag time by a greater percentage in the digesters with a higher total solids content. There was a 17%, 27% and 41% reduction lag time due to biochar addition at total solids contents of 5%, 10% and 20%, respectively. The peak daily methane yield increased by 136% at 10% total solids. There was no significant increase in the peak yield at 5% total solids, while there was a 46% increase at 20% total solids. Real-time PCR analysis confirms the Methanosaetaceae family, which is a key methanogen due to its ability to facilitate direct interspecies electron transfer while attached to biochar, preferentially attaches to biochar. Furthermore, this research shows the attachment of the Methanosaetaceae family, does not decrease with increasing total solids content. A potential negative effect of biochar addition, a reduced volumetric efficiency, can be negated by using a shorter retention time. This new understanding will help to improve predictions of the impact of biochar addition for new digester designs operating in semi-solids and high-solids conditions.
Publisher: Springer Science and Business Media LLC
Date: 21-02-2014
Publisher: MDPI AG
Date: 11-04-2019
DOI: 10.3390/ANTIBIOTICS8020039
Abstract: Infectious diseases remain a major burden in today’s world, causing high mortality rates and significant economic losses, with million deaths per year predicted by 2030. Invasion of host cells by intracellular bacteria poses treatment challenges due to the poor permeation of antimicrobials into the infected cells. To overcome these limitations, mesoporous silica nanoparticles (MSNP) loaded with the antibiotic rif icin were investigated as a nanocarrier system for the treatment of intracellular bacterial infection with specific interest in the influence of particle size on treatment efficiency. An intracellular infection model was established using small colony variants (SCV) of S. aureus in macrophages to systemically evaluate the efficacy of rif icin-loaded MSNP against the pathogen as compared to a rif icin solution. As hypothesized, the superior uptake of MSNP by macrophages resulted in an enhanced treatment efficacy of the encapsulated rif icin as compared to free antibiotic. This study provides a potential platform to improve the performance of currently available antibiotics against intracellular infections.
Publisher: Wiley
Date: 03-03-2006
DOI: 10.1111/J.1365-2958.2006.05079.X
Abstract: In previous studies it has been established that resistance to superoxide by Neisseria gonorrhoeae is dependent on the accumulation of Mn(II) ions involving the ABC transporter, MntABC. A mutant strain lacking the periplasmic binding protein component (MntC) of this transport system is hypersensitive to killing by superoxide anion. In this study the mntC mutant was found to be more sensitive to H2O2 killing than the wild-type. Analysis of regulation of MntC expression revealed that it was de-repressed under low Mn(II) conditions. The N. gonorrhoeae mntABC locus lacks the mntR repressor typically found associated with this locus in other organisms. A search for a candidate regulator of mntABC expression revealed a homologue of PerR, a Mn-dependent peroxide-responsive regulator found in Gram-positive organisms. A perR mutant expressed more MntC protein than wild-type, and expression was independent of Mn(II), consistent with a role for PerR as a repressor of mntABC expression. The PerR regulon of N. gonorrhoeae was defined by microarray analysis and includes ribosomal proteins, TonB-dependent receptors and an alcohol dehydrogenase. Both the mntC and perR mutants had reduced intracellular survival in a human cervical epithelial cell model.
Publisher: Oxford University Press (OUP)
Date: 15-12-2007
DOI: 10.1086/523107
Abstract: A transcriptional regulator, NmlR(sp), has been identified in Streptococcus pneumoniae that is required for defense against nitric oxide (NO) stress. The nmlR(sp) gene is cotranscribed with adhC, which encodes an alcohol dehydrogenase that is able to reduce S-nitrosoglutathione (GSNO) with NADH as reductant. nmlR(sp) and adhC mutants exhibited a reduced level of NADH-GSNO oxidoreductase activity and were more susceptible to killing by NO than were wild-type cells. Comparison of the virulence of wild-type and mutant strains by use of a mouse model system showed that NmlR(sp) and AdhC do not play a key role in the adherence of pneumococci to the nasopharynx in vivo. An intraperitoneal challenge experiment revealed that both NmlR(sp) and AdhC were required for survival in blood. These data identify novel components of a NO defense system in pneumococci that are required for systemic infection.
Publisher: Elsevier BV
Date: 02-2020
DOI: 10.1016/J.MSEC.2019.110220
Abstract: ZnO nanoparticles doped with I and Ag were prepared via a solvothermal method. Characterizations of the as-synthesised s les were carried out using X-ray diffraction, X-ray photoelectron spectroscopy, UV-Vis spectrometry, Photoluminescence, transmission electron microscopy and scanning electron microscopy. The nanoparticles exhibit light absorption for wide spectra from ultra-violet (UV) to visible light. The antimicrobial efficacy was evaluated against Escherichia coli (MG1655) and Staphylococcus aureus (USA300) as models of Gram-negative and Gram-positive microorganisms, respectively. The double-doped nanoparticles demonstrated their potent efficacy against both types of microorganisms and they may have great potential in combating infectious diseases. More importantly, the insights into the mechanisms underlying the antimicrobial effects were revealed: synergistic effect of reactive oxygen species (ROS) generation and Ag
Publisher: Oxford University Press (OUP)
Date: 15-01-2009
DOI: 10.1086/595737
Publisher: American Society for Microbiology
Date: 02-2015
DOI: 10.1128/IAI.02702-14
Abstract: An undetermined feature of Staphylococcus aureus pathogenesis is its persistence and then relapse of disease. This has been explained by its switch to alternative lifestyles, mainly as biofilm or small-colony variants (SCVs). Studying the native characteristics of SCVs has been problematic due to their reversion to the parental lifestyle. We have observed that for a number of S. aureus strains as they switch to an SCV lifestyle, there is the formation of an extracellular matrix. We focused our analysis on one strain, WCH-SK2. For bacterial survival in the host, the combination of low nutrients and the prolonged time frame forms a stress that selects for a specific cell type from the population. In this context, we used steady-state growth conditions with low nutrients and a controlled low growth rate for a prolonged time and with methylglyoxal. These conditions induced S. aureus WCH-SK2 into a stable SCV cell type the cells did not revert after subculturing. Analysis revealed these cells possessed a metabolic and surface profile that was different from those of previously described SCVs or biofilm cells. The extracellular matrix was protein and extracellular DNA but not polysaccharide. The SCV cells induced expression of certain surface proteins (such as Ebh) and synthesis of lantibiotics while downregulating factors that stimulate the immune response (leucocidin, capsule, and carotenoid). Our data reveal cell heterogeneity within an S. aureus population and under conditions that resemble long-term survival in the host have identified a previously unnoticed S. aureus cell type with a distinctive metabolic and molecular profile.
Publisher: Elsevier BV
Date: 2015
DOI: 10.1016/J.MICINF.2014.09.009
Abstract: Staphylococcus aureus is extremely versatile. It has a capacity to persist within its host by switching to the alternative lifestyles of biofilm or Small Colony Variants (SCV). The induction of this switch has been presumed to be in response to stressed conditions, however the environmental basis has not been thoroughly investigated. We assessed the response of numerous strains to chemicals that are present in human host. There were some that induced a biofilm or SCV phenotype and indeed some inducing both lifestyles. This result illustrates the ersity within a population and a strain-specific adaptation to the presence of host-generated stresses.
Publisher: Springer Science and Business Media LLC
Date: 29-07-2004
DOI: 10.1007/S00484-004-0225-3
Abstract: Total (as opposed to culturable) bacterial number counts are reported for four sites in the United Kingdom measured during c aigns over four separate seasons. These are interpreted in relation to simple climatic factors, i.e. temperature, wind speed and wind direction. Temperature has a marked effect at all four sites with data for a rural coastal site conforming best to a simple exponential model. Data for the other rural and urban locations show a baseline similar to that determined at the coastal rural location, but with some very significant positive excursions. The temperature dependence of bacterial number is found to conform to that typical of bacterial growth rates. At the coastal rural location, bacterial numbers normalised for temperature show no dependence on wind speed whilst at the inland sites there is a decrease with increasing wind speed of the form expected for a large area source. Only one site appeared to show a systematic relationship of bacterial concentrations to wind direction that being a site in the suburbs of Birmingham with highest number concentrations observed on a wind sector approaching from the city centre. PCR techniques have been used to identify predominant types of bacteria and results are presented which show that Bacillus was the dominant genus observed at the three inland sites during the winter and summer seasons. Pseudomonas appeared with comparable frequency at certain sites and seasons. There was in general a greater ersity of bacteria at the coastal site than at the inland sites.
Publisher: MDPI AG
Date: 11-05-2023
DOI: 10.3390/ANTIBIOTICS12050895
Abstract: The similarity of commensal Escherichia coli isolated from healthy cattle to antimicrobial-resistant bacteria causing extraintestinal infections in humans is not fully understood. In this study, we used a bioinformatics approach based on whole genome sequencing data to determine the genetic characteristics and phylogenetic relationships among faecal Escherichia coli isolates from beef cattle (n = 37) from a single feedlot in comparison to previously analysed pig faecal (n = 45), poultry extraintestinal (n = 19), and human extraintestinal E. coli isolates (n = 40) from three previous Australian studies. Most beef cattle and pig isolates belonged to E. coli phylogroups A and B1, whereas most avian and human isolates belonged to B2 and D, although a single human extraintestinal isolate belonged to phylogenetic group A and sequence type (ST) 10. The most common E. coli sequence types (STs) included ST10 for beef cattle, ST361 for pig, ST117 for poultry, and ST73 for human isolates. Extended-spectrum and AmpC β-lactamase genes were identified in seven out of thirty-seven (18.9%) beef cattle isolates. The most common plasmid replicons identified were IncFIB (AP001918), followed by IncFII, Col156, and IncX1. The results confirm that feedlot cattle isolates examined in this study represent a reduced risk to human and environmental health with regard to being a source of antimicrobial-resistant E. coli of clinical importance.
Publisher: Elsevier BV
Date: 12-2015
DOI: 10.1016/J.IJMM.2015.09.003
Abstract: Haemophilus influenzae and Streptococcus pneumoniae exist together as common commensals of the healthy human nasopharynx, but both are important aetiological agents of different diseases, including the paediatric disease otitis media. It was recently shown that the formation of a multispecies biofilm of H. influenzae and S. pneumoniae is the cause of chronic forms of otitis media. However, the interactions between the two species are not clearly defined. Using a defined and kinetic analysis, our study has shown that while co-existence of the two species occurs, S. pneumoniae is also able to convert H. influenzae to a non-culturable state. We determined that this process was dependent on growth phase and pH. To analyse the H. influenzae/S. pneumoniae interactions in more depth, we investigated the growth and transcriptional profile in a pH-defined batch culture model, as well as in a growth phase independent flow cell system. Transcriptomics has shown that there are changes in gene expression in each of the species when grown in co-culture, intriguingly inducing the S. pneumoniae bacteriocin transport genes, and phage-associated genes in both species. Importantly, we have shown vast changes in gene expression in a group of S. pneumoniae metabolic genes, including those encoding lactose utilisation, glycerol utilisation and sugar transport proteins we have shown that the expression of these genes depends not only on the presence of H. influenzae, but also on the growth system utilised.
Publisher: American Chemical Society (ACS)
Date: 23-07-2021
Abstract: Antibacterial treatment strategies using functional nanomaterials, such as photodynamic therapy, are urgently required to combat persistent
Publisher: IEEE
Date: 10-2019
Publisher: Elsevier BV
Date: 02-2023
Publisher: American Society for Microbiology
Date: 02-05-2018
Abstract: Periprosthetic joint infection (PJI) is a potentially devastating complication of orthopedic joint replacement surgery. PJI with associated osteomyelitis is particularly problematic and difficult to cure. Whether viable osteocytes, the predominant cell type in mineralized bone tissue, have a role in these infections is not clear, although their involvement might contribute to the difficulty in detecting and clearing PJI. Here, using Staphylococcus aureus , the most common pathogen in PJI, we demonstrate intracellular infection of human-osteocyte-like cells in vitro and S. aureus adaptation by forming quasi-dormant small-colony variants (SCVs). Consistent patterns of host gene expression were observed between in vitro -infected osteocyte-like cultures, an ex vivo human bone infection model, and bone s les obtained from PJI patients. Finally, we confirm S. aureus infection of osteocytes in clinical cases of PJI. Our findings are consistent with osteocyte infection being a feature of human PJI and suggest that this cell type may provide a reservoir for silent or persistent infection. We suggest that elucidating the molecular/cellular mechanism(s) of osteocyte-bacterium interactions will contribute to better understanding of PJI and osteomyelitis, improved pathogen detection, and treatment. IMPORTANCE Periprosthetic joint infections (PJIs) are increasing and are recognized as one of the most common modes of failure of joint replacements. Osteomyelitis arising from PJI is challenging to treat and difficult to cure and increases patient mortality 5-fold. Staphylococcus aureus is the most common pathogen causing PJI. PJI can have subtle symptoms and lie dormant or go undiagnosed for many years, suggesting persistent bacterial infection. Osteocytes, the major bone cell type, reside in bony caves and tunnels, the lacuno-canalicular system. We report here that S. aureus can infect and reside in human osteocytes without causing cell death both experimentally and in bone s les from patients with PJI. We demonstrate that osteocytes respond to infection by the differential regulation of a large number of genes. S. aureus adapts during intracellular infection of osteocytes by adopting the quasi-dormant small-colony variant (SCV) lifestyle, which might contribute to persistent or silent infection. Our findings shed new light on the etiology of PJI and osteomyelitis in general.
Publisher: Wiley
Date: 11-10-2010
DOI: 10.1111/J.1365-2958.2010.07401.X
Abstract: Mycobacterium ulcerans is the causative agent of the debilitating skin disease Buruli ulcer, which is most prevalent in Western and Central Africa. M. ulcerans shares >98% DNA sequence identity with Mycobacterium marinum, however, M. marinum produces granulomatous, but not ulcerative, lesions in humans and animals. Here we report the differential expression of a small heat shock protein (Hsp18) between strains of M. ulcerans (Hsp18(+) ) and M. marinum (Hsp18(-) ) and describe the molecular basis for this difference. We show by gene deletion and GFP reporter assays in M. marinum that a ergently transcribed gene called hspR_2, immediately upstream of hsp18, encodes a MerR-like regulatory protein that represses hsp18 transcription while promoting its own expression. Naturally occurring mutations within a 70 bp segment of the 144 bp hspR_2-hsp18 intergenic region among M. ulcerans strains inhibit hspR_2 transcription and explain the Hsp18(+) phenotype. We also propose a biological role for Hsp18, as we show that this protein significantly enhances bacterial attachment or aggregation during biofilm formation. This study has uncovered a new member of the MerR family of transcriptional regulators and suggests that upregulation of hsp18 expression was an important pathoadaptive response in the evolution of M. ulcerans from a M. marinum-like ancestor.
Publisher: Portland Press Ltd.
Date: 28-02-2017
DOI: 10.1042/EBC20160061
Abstract: Staphylococcus aureus has an incredible ability to survive, either by adapting to environmental conditions or defending against exogenous stress. Although there are certainly important genetic traits, in part this ability is provided by the breadth of modes of growth S. aureus can adopt. It has been proposed that while within their host, S. aureus survives host-generated and therapeutic antimicrobial stress via alternative lifestyles: a persister sub-population, through biofilm growth on host tissue or by growing as small colony variants (SCVs). Key to an understanding of chronic and relapsing S. aureus infections is determining the molecular basis for its switch to these quasi-dormant lifestyles. In a multicellular biofilm, the metabolically quiescent bacterial community additionally produces a highly protective extracellular polymeric substance (EPS). Furthermore, there are bacteria within a biofilm community that have an altered physiology potentially equivalent to persister cells. Recent studies have directly linked the cellular ATP production by persister cells as their key feature and the basis for their tolerance of a range of antibiotics. In clinical settings, SCVs of S. aureus have been observed for many years when cultured, these cells form non-pigmented colonies and are approximately ten times smaller than their counterparts. Various genotypic factors have been identified in attempts to characterize S. aureus SCVs and different environmental stresses have been implicated as important inducers.
Publisher: Elsevier BV
Date: 02-2016
DOI: 10.1016/J.RESMIC.2015.09.008
Abstract: The survival by pathogenic bacteria within the specific conditions of an anatomical niche is critical for their persistence. These conditions include the combination of toxic chemicals, such as reactive oxygen (ROS) and reactive nitrogen species (RNS), with factors relevant to cell growth, such as oxygen. Haemophilus influenzae senses oxygen levels largely through the redox state of the intracellular fumarate-nitrate global regulator (FNR). H. influenzae certainly encounters oxygen levels that fluctuate, but in reality, these would rarely reach a state that results in FNR being fully reduced or oxidized. We were therefore interested in the response of H. influenzae to ROS and RNS at moderately high or low oxygen levels and the corresponding role of FNR. At these levels of oxygen, even though the growth rate of an H. influenzae fnr mutant was similar to wild type, its ROS and RNS tolerance was significantly different. Additionally, the subtle changes in oxygen did alter the whole cell transcriptional profile and this was different between the wild type and fnr mutant strains. It was the changed whole cell profile that impacted on ROS/RNS defence, but surprisingly, the FNR-regulated, anaerobic nitrite reductase (NrfA) continued to be expressed and had a role in this phenotype.
Publisher: American Society for Microbiology
Date: 09-2007
DOI: 10.1128/IAI.00487-07
Abstract: In Haemophilus influenzae Rd KW20, we identified a gene, adhC , which encodes a class III alcohol dehydrogenase (AdhC) and has S -nitrosoglutathione reductase activity. adhC exists on an operon with estD , which encodes an esterase. Divergent to the adhC - estD operon is the Haemophilus influenzae nmlR gene ( nmlR HI ), which encodes a MerR family regulator that is homologous to the Neisseria MerR-like regulator (NmlR). Analysis of an nmlR HI mutant indicated that expression of the adhC - estD operon is regulated by NmlR HI in strain Rd KW20. Chromosomal inactivation of either adhC or nmlR HI resulted in sensitivity to S -nitrosoglutathione and decreased S -nitrosoglutathione reductase activity. Examination of the NmlR HI -AdhC system in the genome sequences of nontypeable H. influenzae strains R2846, R2866, and 86-028NP identified significant variations. The adhC gene of 86-028NP was predicted to be nonfunctional due to a premature stop codon. Polymorphisms in the operator romoter region of R2866 resulted in reduced enzyme activity. This correlated with an increased sensitivity to S -nitrosoglutathione. The adhC - nmlR HI system was examined in thirty-three clinical isolates (both capsular and nontypeable strains). Nucleic acid sequence data showed that only strain 86-028NP contained a premature stop codon. There were some variations in the DNA sequence of the operator romoter region which altered the nmlR HI promoter. However, the clinical isolates still possessed S -nitrosoglutathione reductase activity and showed at least the equivalent ability to grow in the presence of S -nitrosoglutathione as Rd KW20. These data suggest that the nmlR HI - adhC system has a role in the defense against nitrosative stress in Haemophilus influenzae .
Publisher: Oxford University Press (OUP)
Date: 06-2003
Publisher: Microbiology Society
Date: 16-06-2023
DOI: 10.1099/JMM.0.001716
Abstract: Introduction. Uninfected diabetes-related foot ulcer (DFU) progression to diabetes-related foot infection (DFI) is a prevalent complication for patients with diabetes. DFI often progresses to osteomyelitis (DFI-OM). Active (growing) Staphylococcus aureus is the most common pathogen in these infections. There is relapse in 40–60 % of cases even when the initial treatment at the DFI stage apparently clears infection. Hypothesis. S. aureus adopts the quasi-dormant Small Colony Variant (SCV) state during DFU and consequently infection, and when present in DFI cases also permits survival in non-diseased tissues as a reservoir to cause relapse. Aim . The aim of this study was to investigate the bacterial factors that facilitate persistent infections. Methodology. People with diabetes were recruited from two tertiary hospitals. Clinical and bacterial data was taken from 153 patients with diabetes (51 from a control group with no ulcer or infection) and s les taken from 102 patients with foot complications to identify bacterial species and their variant colony types, and then compare the bacterial composition in those with uninfected DFU, DFI and those with DFI-OM, of whom s les were taken both from wounds (DFI-OM/W) and bone (DFI-OM/B). Intracellular, extracellular and proximal ‘healthy’ bone were examined. Results. S. aureus was identified as the most prevalent pathogen in diabetes-related foot pathologies (25 % of all s les). For patients where disease progressed from DFU to DFI-OM, S. aureus was isolated as a ersity of colony types, with increasing numbers of SCVs present. Intracellular (bone) SCVs were found, and even within uninfected bone SCVs were present. Wounds of 24 % of patients with uninfected DFU contained active S. aureus . All patients with a DFI with a wound but not bone infection had previously had S. aureus isolated from an infection (including utation), representing a relapse. Conclusion. The presence of S. aureus SCVs in recalcitrant pathologies highlights their importance in persistent infections through the colonization of reservoirs, such as bone. The survival of these cells in intracellular bone is an important clinical finding supporting in vitro data. Also, there seems to be a link between the genetics of S. aureus found in deeper infections compared to those only found in DFU.
Publisher: MDPI AG
Date: 06-10-2022
DOI: 10.3390/ANI12192690
Abstract: Enterococcus faecium are commensal bacteria inhabiting the gastrointestinal tract of animals and humans and an important cause of drug-resistant nosocomial infections. This longitudinal study aimed to determine whether changes in the antimicrobial resistance (AMR) phenotype and genotype occurred among Enterococcus spp. isolated from cattle rectal s les obtained at the entry to and exit from an Australian feedlot. The s les obtained at the feedlot induction yielded enterococci (104/150 69.3%), speciated as E. hirae (90/104 86.5%), E. faecium (9/104 8.7%), E. mundtii (3/104 2.9%), E. durans, and E. casseliflavus (1/104 1.0% each). AMR was observed to lincomycin (63/104 60.6%), daptomycin (26/104 25.0%), nitrofurantoin (9/104 8.7%), ciprofloxacin (7/104 6.7%), tetracycline (5/104 4.8%), tigecycline (4/104 3.9%), and quinupristin/dalfopristin (3/104 2.9%). From the rectal swab s les collected at the abattoir from the same animals (i.e., the feedlot exit), the enterococci recovery was significantly higher (144/150 96.0%), with a marked shift in species distribution dominated by E. faecium (117/144 81.3%). However, the prevalence of AMR to in idual antimicrobials remained largely static between the entry and exit except for the increased resistance to nitrofurantoin (77/144 53.5%) and quinupristin/dalfopristin (26/144 18.1%). Overall, 13 AMR genes were observed among the 62 E. faecium isolates. These included aac(6′)Ii, aac(6′)-Iid, and ant(6)-Ia (aminoglycosides) eatAv, lnu(G), vat(E), msr(C), and erm(B) (macrolides, lincosamides, and streptogramins) efmA (fluoroquinolones) and tet(45), tet(L), tet(M), and tet(S) (tetracyclines). The results confirm the presence of fluoroquinolone- and streptogramin-resistant enterococci in cattle faeces at the feedlot entry in the absence of antimicrobial selection pressure. E. faecium, exhibiting increased nitrofurantoin resistance, became the dominant Enterococcus spp. during the feeding period.
Publisher: Frontiers Media SA
Date: 28-02-2020
Publisher: Oxford University Press (OUP)
Date: 2011
DOI: 10.1039/C1MT00127B
Abstract: We have identified a novel regulator from the MerR family of transcription factors in the bacterial pathogen Haemophilus influenzae (HI1623 nickel-associated merR-like Regulator--NimR). NimR regulates the expression of a Ni(2+) uptake transporter (NikKLMQO). The promoters for nimR and the nik operon are ergent and overlapping and NimR binds at a site between the promoter elements for nikKLMQO. Expression of this operon requires NimR and depends on Ni(2+). Growth rates of the H. influenzae nimR and nikQ mutants were reduced in chemically defined media compared to the wild type and the mutants were unable to grow in the presence of EDTA. The mutant strains were less tolerant of acidic pH and the wild type Rd KW20 could not tolerate low pH in the presence of fluoramide, a urease specific inhibitor, confirming that both nickel transport and urea hydrolysis are a central process in pH control. H. influenzae nimR and nikQ strains were deficient in urease activity, but this could be specifically restored by the addition of excess Ni(2+). NimR did not directly regulate the expression of urease genes but the activity of urease requires both nimR and nikQ. Purified NimR is a dimer that binds 1 Ni(2+)ion. NimR is the first ex le of a Ni-dependent regulator from the MerR family and targeting a metal ion uptake system it is distinct from NikR the Ni-responsive regulators of the ribbon-helix-helix family.
Publisher: Elsevier BV
Date: 03-2005
DOI: 10.1016/J.BBRC.2005.01.084
Abstract: The cueO gene of Escherichia coli encodes a multi-copper oxidase, which contributes to copper tolerance in this bacterium. It was observed that a cueO mutant was highly sensitive to killing by copper ions when cells were grown on defined minimal media. Copper sensitivity was correlated with accumulation of copper in the mutant strain. Growth of the cueO mutant in the presence of copper could be restored by addition of alent zinc and manganese ions or ferrous iron but not by other first row transition metal ions or magnesium ions. Copper toxicity towards a cueO mutant could also be suppressed by addition of the superoxide quencher 1,2-dihydroxybenzene-3,5-disulfonic acid (tiron), suggesting that a primary cause of copper toxicity is the copper-catalyzed production of superoxide anions in the cytoplasm.
Publisher: Oxford University Press (OUP)
Date: 10-12-2021
Abstract: Factors facilitating the chronicity of otitis media (OM) in children are, to date, not fully understood. An understanding of molecular factors aiding bacterial persistence within the middle ear during OM could reveal pathways required for disease. This study performed a detailed analysis of Streptococcus pneumoniae populations isolated from the nasopharynx and middle ear of one OM case. Isolates were assessed for growth in vitro and infection in a mouse intranasal challenge model. Whole genome sequencing was performed to compare the nasopharyngeal and middle ear isolates. The middle ear isolate displayed a reduced rate of growth and enhanced potential to transit to the middle ear in a murine model. The middle ear population possessed a single nucleotide polymorphism (SNP) in the IgA1 protease gene igA, predicted to render its product non-functional. Allelic exchange mutagenesis of the igA alleles from the genetic variant middle ear and nasopharyngeal isolates was able to reverse the niche-adaptation phenotype in the murine model. These results indicate the potential role of a SNP in the gene encoding the IgA1 protease, in determining S. pneumoniae adaptation to the middle ear during chronic OM. In contrast, a functional IgA1 protease was associated with increased colonisation of the nasopharynx.
Publisher: Elsevier BV
Date: 03-2010
Publisher: American Society for Microbiology
Date: 03-2012
DOI: 10.1128/IAI.06163-11
Abstract: NGO0579 is annotated copA in the Neisseria gonorrhoeae chromosome, suggesting that it encodes a cation-transporting ATPase specific for copper ions. Compared to wild-type cells, a copA mutant was more sensitive to killing by copper ions but not to other transition metals. The mutant also accumulated a greater amount of copper, consistent with the predicted role of CopA as a copper efflux pump. The copA mutant showed a reduced ability to invade and survive within human cervical epithelial cells, although its ability to form a biofilm on the surface of these cells was not significantly different from that of the wild type. In the presence of copper, the copA mutant exhibited increased sensitivity to killing by nitrite or nitric oxide. Therefore, we concluded that copper ion efflux catalyzed by CopA is linked to the nitrosative stress defense system of Neisseria gonorrhoeae . These observations suggest that copper may exert its effects as an antibacterial agent in the innate immune system via an interaction with reactive nitrogen species.
Publisher: American Society for Microbiology
Date: 03-2003
DOI: 10.1128/AAC.47.3.1115-1119.2003
Abstract: Three mer transposons from the Murray collection of preantibiotic enterobacteria show % sequence identity to current isolates. Tn 5073 is most closely related to Tn 5036 and Tn 1696 , and Tn 5074 is most closely related to Tn 5053 . Tn 5075 is most closely related to Tn 21 but lacks integron In2 and is flanked by insertion elements.
Publisher: Springer Science and Business Media LLC
Date: 2012
Publisher: Oxford University Press (OUP)
Date: 2015
DOI: 10.1039/C4MT00245H
Abstract: Nickel acts as a co-factor for a small number of enzymes in bacteria. Urease is one of the two nickel-dependent enzymes that have been identified in Haemophilus influenzae glyoxalase I is the other. However, nickel has been suggested to have roles in H. influenzae that can not attributed to the function of these enzymes. We have previously shown that in the H. influenzae strain Rd KW20 the inability to acquire nickel led to alterations to the cell-type an increased biofilm formation and changes in cell surface properties. Here we report the differences in the genome wide gene expression between Rd KW20 and a strain incapable of importing nickel (nikQ) revealing a link between intracellular nickel levels and genes involved in metabolic pathways, stress responses and genes associated with surface factors such as type IV pili. We have then taken a strain previously shown to use type IV pili both in biofilm formation and for twitching motility (86-028NP) and have shown its homologous genes (NTHI1417-1422 annotated as cobalt transporter, cbiKLMOQ) did import nickel and mutations in this locus had pleiotropic effects correlating to stress response and motility. Compared to wild type cells, the nickel depleted cells were more electronegativity charged, they aggregated and formed a biofilm. Correct intracellular nickel levels were also important for resistance to oxidative stress the nickel depleted cells were more sensitive to oxidative stress. The nickel depleted cells were also non-motile, but the addition specifically of nickel returned these cells to a wild type motility state. We have also analysed the role of nickel uptake in a naturally, urease negative strain (the blood isolate R2866) and depleting intracellular nickel (a nikQ mutant) in this strain effected a similar range of cell functions. These data reveal a role for the capacity to acquire nickel from the environment and for the correct intracellular nickel levels as part of H. influenzae stress response and in signalling for a switch to a sessile bacterial lifestyle.
Publisher: Oxford University Press (OUP)
Date: 10-09-2013
Publisher: Elsevier BV
Date: 03-2003
DOI: 10.1016/S0006-291X(03)00249-3
Abstract: A transcriptional regulator of the MerR family encoded by Bordetella pertussis was characterized in Escherichia coli and in vitro. Uniquely, the regulator responded specifically to Zn(II), Cd(II), and Co(II) and was named ZccR. Gel shift assays confirmed that ZccR binds to an adjacent ergent promoter possessing an elongated spacer region of 19bp between the -10 and -35 elements, and that Zn(II), Co(II), and Cd(II) reduced the protein affinity for DNA. Site-directed mutagenesis of four cysteine and six histidine residues of ZccR showed that the cysteine residues at positions 77, 112, and 122, conserved in many of the metal-responsive MerR-like regulators, were essential for induction. Mutagenesis of the histidine residues (positions 73, 87, 90, 126, 140, and 142) revealed that histidine residues at 90, 140, and 142 were required for full induction by all three metals.
Publisher: Elsevier BV
Date: 02-2020
DOI: 10.1016/J.BIORTECH.2019.122457
Abstract: This study characterises the effect of biochar (pyrolysed biomass) produced from wood pellets, wheat straw and sheep manure on high-solids anaerobic digestion (HSAD) of poultry litter. Also, pre-loading biochar with microorganisms before addition to HSADs was investigated. The addition of wood pellet biochar provides a 32% increase to the methane yield compared with control digesters. The addition of biochar produced from either wheat straw or sheep manure has detrimental effects on digester performance compared with controls. The addition of wood pellet biochar pre-loaded by placing it in a high-solids digester for 90 days provides a 69% increase in the total methane yield, 44% increase in the peak daily methane yield and a 33% reduction in the lag time compared with controls. This study highlighted a need for careful selection of parent material for biochar production and, for the first time, the opportunities to re-use wood pellet biochar for further improvements.
Publisher: Oxford University Press (OUP)
Date: 02-2002
DOI: 10.1046/J.1365-2672.2002.01529.X
Abstract: The aim of this study was to identify, clone and characterize the second amylase of Aeromonas hydrophila JMP636, AmyB, and to compare it to AmyA. The amylase activity of A. hydrophila JMP636 is encoded by multiple genes. A second genetically distinct amylase gene, amyB, has been cloned and expressed from its own promoter in Escherichia coli. AmyB is a large alpha-amylase of 668 amino acids. Outside the conserved domains of alpha-amylases there is limited sequence relationship between the two alpha-amylases of A. hydrophila JMP636 AmyA and AmyB. Significant (80%) similarity exists between amyB and an alpha-amylase of A. hydrophila strain MCC-1. Differences in either the functional properties or activity under different environmental conditions as possible explanations for multiple copies of amylases in JMP636 is less likely after an examination of several physical properties, with each of the properties being very similar for both enzymes (optimal pH and temperature, heat instability). However the reaction end products and substrate specificity did vary enough to give a possible reason for the two enzymes being present. Both enzymes were confirmed to be alpha-type amylases. AmyB has been isolated, characterized and then compared to AmyA. The amylase phenotype is rarely encoded by more than one enzyme within one strain, this study therefore allows the better understanding of the unusual amylase production by A. hydrophila.
Publisher: Oxford University Press (OUP)
Date: 11-2002
DOI: 10.1046/J.1365-2672.2002.01750.X
Abstract: The physiological examination of amylase production by Aeromonas hydrophila JMP636 and identification of the mechanism of regulation. Aeromonas hydrophila JMP636 was grown with single, then dual carbon sources the growth cycle was followed and amylase activity throughout was monitored. The levels of cAMP, a known secondary messenger for the regulatory gene crp, were also examined. Amylase activity was regulated by catabolite repression. Physiological studies revealed that JMP636 exhibited both diauxic growth, with two carbon sources, and the 'acid toxicity' effect on glucose. The crp gene was cloned, expressed and inactivated from the JMP636 chromosome. Catabolite repression of amylase production and the 'acid toxicity' effect both require crp and were linked to cAMP levels. Regulation of amylase production was predicted to follow the model CRP-mediated cAMP-dependent Escherichia coli catabolite regulation system. This work provides an understanding of the physiology of the opportunistic pathogen Aer. hydrophila through identification of the mechanism of catabolite repression of amylase production and the existence of crp within this cell. It also provides a broader knowledge of global gene regulation and suggests regulatory mechanisms of other Aer. hydrophila gene/s.
Publisher: Frontiers Media SA
Date: 26-12-2016
Publisher: No publisher found
Date: 2023
Publisher: Elsevier BV
Date: 02-2017
DOI: 10.1016/J.JOEN.2017.08.025
Abstract: Extracellular material (ECM) surrounding Enterococcus faecalis may play a role in increasing resistance to environmental stresses. Our aim was to determine ECM levels in response to subminimal inhibitory concentrations of sodium hypochlorite (sub-MIC/NaOCl) or anaerobic growth and determine the impact on biofilm development. From 37 E. faecalis clinical strains, 19 were selected according to their biofilm-producing ability by using a crystal violet biofilm assay: 10 strong, 4 intermediate, and 5 non-biofilm producers. Biofilm assays were subsequently performed on all strains when subjected to sub-MIC/NaOCl. All strains were evaluated for ECM production under aerobic and anaerobic conditions and with sub-MIC/NaOCl. ECM production was assessed by using scanning electron microscopy. Double-blinded independent assessors were used to score levels of ECM production. The esp gene was detected by using polymerase chain reaction. Gelatinase activity was determined by using Todd-Hewitt and gelatin agar. In aerobic conditions, ECM was expressed in all strains. In the presence of sub-MIC/NaOCl, of the 10 strong biofilm producers, 5 increased their ECM production, and 4 showed increased biofilm growth. Two strains had less ECM production and showed decreased biofilm growth. One isolate demonstrated no observable changes. Most non-biofilm producers demonstrated no observable differences in ECM production, although 1 strain increased biofilm growth. ECM production in anaerobic conditions was highly variable. The esp gene (n = 15) and gelatinase activity (n = 7) were evident among the isolates. Clonal ersity among strains of E. faecalis suggests that some strong biofilm producers can upregulate ECM production and increase biofilm growth in response to sub-MIC/NaOCl.
Publisher: MDPI AG
Date: 31-08-2022
DOI: 10.3390/ANI12172256
Abstract: This study investigated the antimicrobial resistance (AMR) profile of fecal Escherichia coli isolates from beef cattle (n = 150) at entry and exit from an Australian feedlot. S le plating on MacConkey agar and Brilliance ESBL agar differentiated generic from extended-spectrum β-lactamase (ESBL)-producing E. coli, respectively. Resistance profiles were determined by minimum inhibitory concentration (MIC) testing and further analyzed by whole-genome sequencing (WGS). At entry, the prevalence of antimicrobial resistance to amoxicillin/clavulanic acid, icillin, streptomycin, and trimethoprim/sulfamethoxazole was very low (0.7%, each). At the exit, the resistance prevalence was moderate to tetracycline (17.8%) and low to icillin (5.4%), streptomycin (4.7%), and sulfisoxazole (3.9%). The most common AMR genes observed in phenotypically resistant isolates were tet(B) (43.2%), aph(3″)-Ib and aph(6)-Id (32.4%), blaTEM-1B, and sul2 (24.3%, each), which are responsible for resistance to tetracyclines, aminoglycosides, β-lactams, and sulfonamides, respectively. The ESBL-producing E. coli were recovered from one s le (0.7%) obtained at entry and six s les (4.0%) at the exit. The ESBL-producing E. coli harbored blaTEM (29.7%), blaCTX m(13.5%), and blaCMY (5.4%). The resistance phenotypes were highly correlated with resistance genotypes (r ≥ 0.85: p 0.05). This study demonstrated that E. coli isolated from feedlot beef cattle can harbour AMR genes, but the low incidence of medically important resistance reflected the prudent antimicrobial use in the Australian industry.
Publisher: MDPI AG
Date: 07-12-2022
Abstract: In the original publication [...]
Publisher: Public Library of Science (PLoS)
Date: 02-02-2017
Publisher: Frontiers Media SA
Date: 03-11-2021
DOI: 10.3389/FCIMB.2021.781022
Abstract: Infectious osteomyelitis associated with periprosthetic joint infections is often recalcitrant to treatment and has a high rate of recurrence. In the case of Staphylococcus aureus , the most common pathogen in all forms of osteomyelitis, this may be attributed in part to residual intracellular infection of host cells, yet this is not generally considered in the treatment strategy. Osteocytes represent a unique cell type in this context due to their abundance, their formation of a syncytium throughout the bone that could facilitate bacterial spread and their relative inaccessibility to professional immune cells. As such, there is potential value in studying the host-pathogen interactions in the context of this cell type in a replicable and scalable in vitro model. Here, we examined the utility of the human osteosarcoma cell line SaOS2 differentiated to an osteocyte-like stage (SaOS2-OY) as an intracellular infection model for S. aureus . We demonstrate that S. aureus is capable of generating stable intracellular infections in SaOS2-OY cells but not in undifferentiated, osteoblast-like SaOS2 cells (SaOS2-OB). In SaOS2-OY cells, S. aureus transitioned towards a quasi-dormant small colony variant (SCV) growth phenotype over a 15-day post-infection period. The infected cells exhibited changes in the expression of key immunomodulatory mediators that are consistent with the infection response of primary osteocytes. Thus, SaOS2-OY is an appropriate cell line model that may be predictive of the interactions between S. aureus and human osteocytes, and this will be useful for studying mechanisms of persistence and for testing the efficacy of potential antimicrobial strategies.
Publisher: Elsevier BV
Date: 04-2013
DOI: 10.1016/J.IJMM.2013.02.012
Abstract: Of the known proteins which use nickel as a co-factor, Haemophilus influenzae contains only urease and glyoxalase I (gloA). We have recently reported that this pathogen harbours a unique nickel uptake system (nikKLMQO-nimR). Unusually, the disruption of the nickel uptake system (nikQ or nimR mutants) resulted in cells that aggregated and formed an increased biofilm compared to the wild type cells. Using a gloA mutant strain and urease-specific inhibitor we showed that this phenotype is not due to the loss-of-function of these enzymes. By generating H. influenzae "resting cells" which are enzymatically inactive but maintain their structural integrity we have shown that the cell aggregation in the nikQ/nimR mutants is not due to the loss of enzymatic function. The nikQ mutant was unable to accumulate nickel but the addition of excess nickel did restore intracellular nickel levels and this resulted in the nikQ mutant returning to the wild type "free-living" phenotype cells with no aggregation and no biofilm formation. We used a range of techniques which showed that the nikQ mutant possesses changes to its cell surface properties. The mutant was more negatively charged than wild type cells as well as being more hydrophobic. Analysis of the outer membrane constituents showed that there were molecular differences. Although the nikQ mutant appears to grow the same as its wild type cell we have shown that there is a change in the "lifestyle" of these nickel limited cells and this induces changes to the surface of the cell to promote cell-cell aggregation and biofilm formation.
Publisher: American Society for Microbiology
Date: 08-2010
DOI: 10.1128/JB.00383-10
Abstract: The NmlR sp transcription factor of Streptococcus pneumoniae is shown to induce adhC (alcohol dehydrogenase) expression in the presence of both formaldehyde and methylglyoxal. nmlR sp and adhC mutant strains display altered and opposite aerobic growth phenotypes. The nmlR sp strain exhibits increased resistance to high oxygen tension, attributable to decreased H 2 O 2 production, which correlated with downregulation of carbamoyl phosphate synthase ( carB ). This indicates a possible role for AdhC in aldehyde metabolism and a broader role for NmlR sp in the regulation of carbon metabolism.
Publisher: MDPI AG
Date: 21-08-2023
DOI: 10.3390/ANI13162684
Abstract: There was an error in the original publication [...]
Publisher: Wiley
Date: 19-08-2005
DOI: 10.1111/J.1365-2958.2005.04773.X
Abstract: A MerR-like regulator (NmlR -Neisseria merR-like Regulator) identified in the Neisseria gonorrhoeae genome lacks the conserved cysteines known to bind metal ions in characterized proteins of this family. Phylogenetic analysis indicates that NmlR defines a subfamily of MerR-like transcription factors with a distinctive pattern of conserved cysteines within their primary structure. NmlR regulates itself and three other genes in N. gonorrhoeae encoding a glutathione-dependent dehydrogenase (AdhC), a CPx-type ATPase (CopA) and a thioredoxin reductase (TrxB). An nmlR mutant lacked the ability to survive oxidative stress induced by diamide and cumene hydroperoxide. It also had > 50-fold lower NADH-S-nitrosoglutathione oxidoreductase activity consistent with a role for AdhC in protection against nitric oxide stress. The upstream sequences of the NmlR regulated genes contained typical MerR-like operator romoter arrangements consisting of a dyad symmetry located between the -35 and -10 elements of the target genes. The NmlR target operator romoters were cloned into a beta-galactosidase reporter system and promoter activity was repressed by the introduction of NmlR in trans. Promoter activity was activated by NmlR in the presence of diamide. Under metal depleted conditions NmlR did not repress P(AdhC) (or P(CopA)) promoter activity, but this was reversed in the presence of Zn(II), indicating repression was Zn(II)-dependent. Analysis of mutated promoters lacking the dyad symmetry revealed constitutive promoter activity which was independent of NmlR. Gel shift assays further confirmed that NmlR bound to the target promoters possessing the dyad symmetry. Site-directed mutagenesis of the four NmlR cysteine residues revealed that they were essential for activation of gene expression by NmlR.
Publisher: European Cells and Materials
Date: 08-10-2021
DOI: 10.22203/ECM.V042A19
Abstract: Osteomyelitis associated with periprosthetic joint infection (PJI) signals a chronic infection and the need for revision surgery. An osteomyelitic bone exhibits distinct morphological features, including evidence for osteolysis and an accelerated bone remodelling into poorly organised, poor-quality bone. In addition to immune cells, various bone cell-types have been implicated in the pathology. The present study sought to determine the types of bone-cell activities in human PJI bones. Acetabular biopsies from peri-implant bone from patients undergoing revision total hip replacement (THR) for chronic PJI (with several identified pathogens) as well as control bone from the same patients and from patients undergoing primary THR were analysed. Histological analysis confirmed that PJI bone presented increased osteoclastic activity compared to control bone. Analysis of osteocyte parameters showed no differences in osteocyte lacunar area between the acetabular bone taken from PJI patients or primary THR controls. Analysis of bone matrix composition using Masson’s trichrome staining and second-harmonic generation microscopy revealed widespread lack of mature collagen, commonly surrounding osteocytes, in PJI bone. Increased expression of known collagenases, such as matrix metallopeptidase (MMP) 13, MMP1 and cathepsin K (CTSK), was measured in infected bone compared to non-infected bone. Human bone and cultured osteocyte-like cells experimentally exposed to Staphylococcus aureus exhibited strongly upregulated expression of MMP1, MMP3 and MMP13 compared to non-exposed controls. In conclusion, the study identified previously unrecognised bone-matrix changes in PJI caused by multiple organisms deriving from osteocytes. Histological examination of bone collagen composition may provide a useful adjunct diagnostic measure of PJI.
Publisher: MDPI AG
Date: 17-02-2023
DOI: 10.3390/ANTIBIOTICS12020406
Abstract: The ability of Staphylococcus aureus to colonise different niches across the human body is linked to an adaptable metabolic capability, as well as its ability to persist within specific tissues despite adverse conditions. In many cases, as S. aureus proliferates within an anatomical niche, there is an associated pathology. The immune response, together with medical interventions such as antibiotics, often removes the S. aureus cells that are causing this disease. However, a common issue in S. aureus infections is a relapse of disease. Within infected tissue, S. aureus exists as a population of cells, and it adopts a ersity of cell types. In evolutionary biology, the concept of “bet-hedging” has established that even in positive conditions, there are members that arise within a population that would be present as non-beneficial, but if those conditions change, these traits could allow survival. For S. aureus, some of these cells within an infection have a reduced fitness, are not rapidly proliferating or are the cause of an active host response and disease, but these do remain even after the disease seems to have been cleared. This is true for persistence against immune responses but also as a continual presence in spite of antibiotic treatment. We propose that the constant arousal of suboptimal populations at any timepoint is a key strategy for S. aureus long-term infection and survival. Thus, understanding the molecular basis for this feature could be instrumental to combat persistent infections.
Location: United Kingdom of Great Britain and Northern Ireland
No related grants have been discovered for stephen kidd.