Characterization of staphylococcal small colony variants and their pathogenic role in biomaterial-related infections with special reference to staphylococcus epidermidis

There are many surgical implanted devices in current use and all are prone to biomaterial-related infections (BRI) associated with staphylococcal biofilm formation. BRI are usually associated with S. epidermidis or S. Aureus and are characterized by treatment failure and chronicity resulting in reoperation, removal of the implant, and loss of function or death. Staphylococcal small colony variants (SCVs) may be generated by exposure to sublethal concentrations of antibiotics or nutrient limitation which may occur in biofilms. Although the characteristics of S. aureus SCVs have been well studied, little information on SCVs of S. epidermidis and their potential role in BRI is currently available. This study was designed to investigate the biochemical and phenotypic characteristics of S. epidermidis SCVs to further identify characteristics which may contribute to their ability to cause these increasingly important infections. Exposure to two to four times the gentamicin MIC led to the emergence of stable S. epidermidis SCVs, and the ability to produce SCVs was strain dependent. These variants were isogenic by PFGE and less immunogenic by western blotting, and SDS-PAGE analysis of whole cell preparations and cell wall fractions showed altered protein profiles when compared to wild type strains. S epidermidis SCVs were resistant to aminoglycosides such as amikacin and/or netilmicin and they were thiamine and/or menadione auxotrophs. Chemiluminescence assays showed a decreased ATP content reflecting the deficiency in electron transport systems which results in a growth rate – all characteristics similar to those of S. aureus SCVs. Analysis of virulence factor production indicated that S. epidermidis SCVs showed increased lipolytic and proteolytic activity when compared to those of S. aureus. Some S. epidermidis SCVs showed phase variation in exopolysaccharide production which enabled them to be more adherent to uncoated plastic -a property that may also be important for the later stages of development of biofilms. Invasion assays demonstrated that some S. epidermidis and S. aureus SCVs were internalised by HUVECs by a receptor-mediated mechanism which differed from that of the wild type strains. Interaction of staphylococci with HUVECs induced cytokine production but SCVs stimulated production of IL1, IL-6 and IL-8 at lower concentrations than their related wild type parents in the first 6 hours of co-incubation. SCVs were also less damaging to the HUVEC cell line after 24 hours when compared to wild type strains. This study supports the suggestion that a switch to the S. epidermidis SCV phenotype could be a mechanism exploited by the wild type strains to facilitate their survival inside the host. The chronicity and increased antibiotic resistance associated with BRI could in part, be explained by the characteristics of SCVs identified in this study. In particular the ability to survive intracellularly combined with reduced immunogenicity and resulting decreased cytokine production, may contribute to persistence of infection. Although SCVs are resistant to some antibiotics, surviving intracellularly may further protect staphylococci from other drugs which are unable to enter mammalian cells. Resistance may be further enhanced for some strains in biofilms where enhanced polysaccharide production may also limit antibiotic access

Characterization of staphylococcal small colony variants and their pathogenic role in biomaterial-related infections with special reference to staphylococcus epidermidis

There are many surgical implanted devices in current use and all are prone to biomaterial-related infections (BRI) associated with staphylococcal biofilm formation. BRI are usually associated with S. epidermidis or S. Aureus and are characterized by treatment failure and chronicity resulting in reoperation, removal of the implant, and loss of function or death. Staphylococcal small colony variants (SCVs) may be generated by exposure to sublethal concentrations of antibiotics or nutrient limitation which may occur in biofilms. Although the characteristics of S. aureus SCVs have been well studied, little information on SCVs of S. epidermidis and their potential role in BRI is currently available. This study was designed to investigate the biochemical and phenotypic characteristics of S. epidermidis SCVs to further identify characteristics which may contribute to their ability to cause these increasingly important infections. Exposure to two to four times the gentamicin MIC led to the emergence of stable S. epidermidis SCVs, and the ability to produce SCVs was strain dependent. These variants were isogenic by PFGE and less immunogenic by western blotting, and SDS-PAGE analysis of whole cell preparations and cell wall fractions showed altered protein profiles when compared to wild type strains. S epidermidis SCVs were resistant to aminoglycosides such as amikacin and/or netilmicin and they were thiamine and/or menadione auxotrophs. Chemiluminescence assays showed a decreased ATP content reflecting the deficiency in electron transport systems which results in a growth rate – all characteristics similar to those of S. aureus SCVs. Analysis of virulence factor production indicated that S. epidermidis SCVs showed increased lipolytic and proteolytic activity when compared to those of S. aureus. Some S. epidermidis SCVs showed phase variation in exopolysaccharide production which enabled them to be more adherent to uncoated plastic -a property that may also be important for the later stages of development of biofilms. Invasion assays demonstrated that some S. epidermidis and S. aureus SCVs were internalised by HUVECs by a receptor-mediated mechanism which differed from that of the wild type strains. Interaction of staphylococci with HUVECs induced cytokine production but SCVs stimulated production of IL1, IL-6 and IL-8 at lower concentrations than their related wild type parents in the first 6 hours of co-incubation. SCVs were also less damaging to the HUVEC cell line after 24 hours when compared to wild type strains. This study supports the suggestion that a switch to the S. epidermidis SCV phenotype could be a mechanism exploited by the wild type strains to facilitate their survival inside the host. The chronicity and increased antibiotic resistance associated with BRI could in part, be explained by the characteristics of SCVs identified in this study. In particular the ability to survive intracellularly combined with reduced immunogenicity and resulting decreased cytokine production, may contribute to persistence of infection. Although SCVs are resistant to some antibiotics, surviving intracellularly may further protect staphylococci from other drugs which are unable to enter mammalian cells. Resistance may be further enhanced for some strains in biofilms where enhanced polysaccharide production may also limit antibiotic access

Prevalence and phase variable expression status of two autotransporters, NalP and MspA, in carriage and disease isolates of Neisseria meningitidis.

Neisseria meningitidis is a human nasopharyngeal commensal capable of causing life-threatening septicemia and meningitis. Many meningococcal surface structures, including the autotransporter proteins NalP and MspA, are subject to phase variation (PV) due to the presence of homopolymeric tracts within their coding sequences. The functions of MspA are unknown. NalP proteolytically cleaves several surface-located virulence factors including the 4CMenB antigen NhbA. Therefore, NalP is a phase-variable regulator of the meningococcal outer membrane and secretome whose expression may reduce isolate susceptibility to 4CMenB-induced immune responses. To improve our understanding of the contributions of MspA and NalP to meningococcal-host interactions, their distribution and phase-variable expression status was studied in epidemiologically relevant samples, including 127 carriage and 514 invasive isolates representative of multiple clonal complexes and serogroups. Prevalence estimates of >98% and >88% were obtained for mspA and nalP, respectively, with no significant differences in their frequencies in disease versus carriage isolates. 16% of serogroup B (MenB) invasive isolates, predominately from clonal complexes ST-269 and ST-461, lacked nalP. Deletion of nalP often resulted from recombination events between flanking repetitive elements. PolyC tract lengths ranged from 6-15 bp in nalP and 6-14 bp in mspA. In an examination of PV status, 58.8% of carriage, and 40.1% of invasive nalP-positive MenB isolates were nalP phase ON. The frequency of this phenotype was not significantly different in serogroup Y (MenY) carriage strains, but was significantly higher in invasive MenY strains (86.3%; p<0.0001). Approximately 90% of MenB carriage and invasive isolates were mspA phase ON; significantly more than MenY carriage (32.7%) or invasive (13.7%) isolates. This differential expression resulted from different mode mspA tract lengths between the serogroups. Our data indicates a differential requirement for NalP and MspA expression in MenB and MenY strains and is a step towards understanding the contributions of phase-variable loci to meningococcal biology

Characterization of staphylococcal small colony variants and their pathogenic role in biomaterial-related infections with special reference to Staphylococcus epidermidis

There are many surgical implanted devices in current use and all are prone to biomaterial-related infections (BRI) associated with staphylococcal biofilm formation. BRI are usually associated with S. epidermidis or S. Aureus and are characterized by treatment failure and chronicity resulting in reoperation, removal of the implant, and loss of function or death. Staphylococcal small colony variants (SCVs) may be generated by exposure to sublethal concentrations of antibiotics or nutrient limitation which may occur in biofilms. Although the characteristics of S. aureus SCVs have been well studied, little information on SCVs of S. epidermidis and their potential role in BRI is currently available. This study was designed to investigate the biochemical and phenotypic characteristics of S. epidermidis SCVs to further identify characteristics which may contribute to their ability to cause these increasingly important infections. Exposure to two to four times the gentamicin MIC led to the emergence of stable S. epidermidis SCVs, and the ability to produce SCVs was strain dependent. These variants were isogenic by PFGE and less immunogenic by western blotting, and SDS-PAGE analysis of whole cell preparations and cell wall fractions showed altered protein profiles when compared to wild type strains. S epidermidis SCVs were resistant to aminoglycosides such as amikacin and/or netilmicin and they were thiamine and/or menadione auxotrophs. Chemiluminescence assays showed a decreased ATP content reflecting the deficiency in electron transport systems which results in a growth rate – all characteristics similar to those of S. aureus SCVs. Analysis of virulence factor production indicated that S. epidermidis SCVs showed increased lipolytic and proteolytic activity when compared to those of S. aureus. Some S. epidermidis SCVs showed phase variation in exopolysaccharide production which enabled them to be more adherent to uncoated plastic -a property that may also be important for the later stages of development of biofilms. Invasion assays demonstrated that some S. epidermidis and S. aureus SCVs were internalised by HUVECs by a receptor-mediated mechanism which differed from that of the wild type strains. Interaction of staphylococci with HUVECs induced cytokine production but SCVs stimulated production of IL1, IL-6 and IL-8 at lower concentrations than their related wild type parents in the first 6 hours of co-incubation. SCVs were also less damaging to the HUVEC cell line after 24 hours when compared to wild type strains. This study supports the suggestion that a switch to the S. epidermidis SCV phenotype could be a mechanism exploited by the wild type strains to facilitate their survival inside the host. The chronicity and increased antibiotic resistance associated with BRI could in part, be explained by the characteristics of SCVs identified in this study. In particular the ability to survive intracellularly combined with reduced immunogenicity and resulting decreased cytokine production, may contribute to persistence of infection. Although SCVs are resistant to some antibiotics, surviving intracellularly may further protect staphylococci from other drugs which are unable to enter mammalian cells. Resistance may be further enhanced for some strains in biofilms where enhanced polysaccharide production may also limit antibiotic access.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Synthesis, characterization, and antimicrobial activity of Schiff bases derived from benzaldehydes and 3,3′-diaminodipropylamine

Six Schiff bases were prepared by reacting 3,3′-diaminodipropylamine with different benzaldehyde derivatives. The structures of these compounds were confirmed through different spectroscopic methods such as 1H-NMR, 13C-NMR and mass spectrometry. The prepared compounds were evaluated in vitro for their antimicrobial activity against a number of pathogenic Gram-positive and Gram-negative bacteria and Candida by the twofold serial dilution method. These compounds showed bacteriostatic rather than bactericidal activities against Gram positive and Gram-negative bacteria. In addition, compound 3c exhibited significant anticandida activity with an MIC of 24 μg/ml and is, therefore, considered as a promising and potential antifungal agent; further modification can be done on the structure of the compound for a better drug candidate in the future

Low covid-19 vaccine acceptance is correlated with conspiracy beliefs among university students in Jordan

Vaccination to prevent coronavirus disease 2019 (COVID-19) emerged as a promising measure to overcome the negative consequences of the pandemic. Since university students could be considered a knowledgeable group, this study aimed to evaluate COVID-19 vaccine acceptance among this group in Jordan. Additionally, we aimed to examine the association between vaccine conspiracy beliefs and vaccine hesitancy. We used an online survey conducted in January 2021 with a chain-referral sampling approach. Conspiracy beliefs were evaluated using the validated Vaccine Conspiracy Belief Scale (VCBS), with higher scores implying embrace of conspiracies. A total of 1106 respondents completed the survey with female predominance (n = 802, 72.5%). The intention to get COVID-19 vaccines was low: 34.9% (yes) compared to 39.6% (no) and 25.5% (maybe). Higher rates of COVID-19 vaccine acceptance were seen among males (42.1%) and students at Health Schools (43.5%). A Low rate of influenza vaccine acceptance was seen as well (28.8%), in addition to 18.6% of respondents being anti-vaccination altogether. A significantly higher VCBS score was correlated with reluctance to get the vaccine (p <0.001). Dependence on social media platforms was significantly associated with lower intention to get COVID-19 vaccines (19.8%) compared to dependence on medical doctors, scientists, and scientific journals (47.2%, p <0.001). The results of this study showed the high prevalence of COVID-19 vaccine hesitancy and its association with conspiracy beliefs among university students in Jordan. The implementation of targeted actions to increase the awareness of such a group is highly recommended. This includes educational programs to disman-tle vaccine conspiracy beliefs and awareness campaigns to build recognition of the safety and efficacy of COVID-19 vaccines

Distribution of phase ON and OFF <i>mspA</i> genes in carriage and disease isolates by serogroup.

a<p>Statistically significant difference in the frequency of the <i>mspA</i> phase ON phenotype between MenB and MenY carriage isolates (<i>p<</i>0.0001).</p>b<p>Statistically significant difference in the frequency of the <i>mspA</i> phase ON phenotype between MenB and MenY invasive isolates (<i>p<</i>0.0001).</p>c<p>Includes 15 strains from ST-11 which harbor a mutation in <i>mspA</i> interrupting the gene. All 15 strains were phase OFF at the polyC tract.</p

Genetic arrangement of the <i>nalP</i> locus in <i>N. meningitidis</i> isolates.

<p>Unidirectional arrows represent ORFs. Repetitive elements are represented by different symbols. dRS3 consensus sequence: ATTCCCNNNNNNNNGGGAAT; REP4 consensus sequence: AAGACCGTCGGGCATCTGCAGCCGTC. Other repetitive sequences are not shown for clarity. Note that the figure is not drawn to scale but is a representation of the various repeat elements and genes present at the <i>nalP</i> locus in <i>N. meningitidis</i> MC58 and four <i>nalP</i>-negative carriage isolates representing Δ<i>nalP</i>1, Δ<i>nalP</i>2, Δ<i>nalP</i>3 and Δ<i>nalP</i>4 classes.</p