scholarly journals Impact of Elements Containing Glycopeptide Resistance Genes on Expression of Virulence in Enterococcus faecalis Peritonitis: a Pilot Study with Rats

2003 ◽  
Vol 47 (5) ◽  
pp. 1560-1564 ◽  
Author(s):  
G. Plantefeve ◽  
H. Dupont ◽  
V. Hubert ◽  
L. Garry ◽  
C. Poüs ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Resistance Genes ◽  
Peritoneal Fluid ◽  
Type Strain ◽  
Wild Type Strain ◽  
Wild Type ◽  
Glycopeptide Resistance ◽  
Necrosis Factor Alpha ◽  
Cell Counts ◽  
Factor Alpha

ABSTRACT The relationship between virulence and chromosomal elements containing glycopeptide resistance genes was experimentally assessed for two transconjugant strains of Enterococcus faecalis (VanA and VanB phenotypes) and compared to that for a susceptible wild-type strain. Microbiologic and inflammatory effects were assessed in a polymicrobial rat model of peritonitis. Mean peritoneal enterococcus concentrations ± standard deviations at day 1 were 2.1 ± 1.9, 1.3 ± 1.1, and 1.7 ± 2.0 log10 CFU/ml for susceptible, VanA, and VanB strains, respectively (P < 0.05). At day 3 also there were lower concentrations of glycopeptide-resistant enterococcal strains in peritoneal fluid (3.2 ± 3.4, 1.8 ± 1.8, and 2.1 ± 2.4 log10 CFU/ml for susceptible, VanA, and VanB strains, respectively [P < 0.05]). Transconjugant glycopeptide-resistant strains were associated with increased peritoneal cell counts at the different evaluation times of the experiment (P < 0.001). Plasma α1-acid glycoprotein concentrations were lower in the presence of the susceptible strain (667 ± 189 mg/liter) than in the presence of the VanA or VanB strain (1,193 ± 419 or 1,210 ± 404 mg/liter, respectively [P < 0.05]), while concentrations of tumor necrosis factor alpha and interleukin-6 in peritoneal fluid remained similar for the strains. These results suggest a trend toward variation of virulence of transconjugant strains compared to the wild-type strain in this peritonitis model.

scholarly journals Disruption of the Genes for ClpXP Protease in Salmonella enterica Serovar Typhimurium Results in Persistent Infection in Mice, and Development of Persistence Requires Endogenous Gamma Interferon and Tumor Necrosis Factor Alpha

2001 ◽  
Vol 69 (5) ◽  
pp. 3164-3174 ◽  
Author(s):  
Tomoko Yamamoto ◽  
Hiroshi Sashinami ◽  
Akiko Takaya ◽  
Toshifumi Tomoyasu ◽  
Hidenori Matsui ◽  
...  
Keyword(s):  
Tumor Necrosis Factor ◽  
Type Strain ◽  
Tumor Necrosis ◽  
Wild Type Strain ◽  
Wild Type ◽  
Necrosis Factor Alpha ◽  
Serovar Typhimurium ◽  
Factor Alpha ◽  
Deficient Mice ◽  
Necrosis Factor

ABSTRACT The enteric pathogen Salmonella enterica serovar Typhimurium, similar to other facultative intracellular pathogens, has been shown to respond to the hostile conditions inside macrophages of the host organism by producing a set of stress proteins that are also induced by various environmental stresses. The stress-induced ClpXP protease is a member of the ATP-dependent proteases, which are known to be responsible for more than 90% of all proteolysis inEscherichia coli. To investigate the contribution of the ClpXP protease to the virulence of serovar Typhimurium we initially cloned the clpP and clpX operon from the pathogenic strain serovar Typhimurium χ3306 and then created insertional mutations in the clpP and/or clpXgene. The ΔclpP and ΔclpX mutants were used to inoculate BALB/c mice by either the intraperitoneal or the oral route and found to be limited in their ability to colonize organs of the lymphatic system and to cause systemic disease in the host. A variety of experiments were performed to determine the possible reasons for the loss of virulence. An oxygen-dependent killing assay using hydrogen peroxide and paraquat (a superoxide anion generator) and a serum killing assay using murine serum demonstrated that all of the serovar Typhimurium ΔclpP and ΔclpX mutants were as resistant to these killing mechanisms as the wild-type strain. On the other hand, the macrophage survival assay revealed that all these mutants were more sensitive to the intracellular environment than the wild-type strain and were unable to grow or survive within peritoneal macrophages of BALB/c mice. In addition, it was revealed that the serovar Typhimurium ClpXP-depleted mutant was not completely cleared but found to persist at low levels within spleens and livers of mice. Interferon gamma-deficient mice and tumor necrosis factor alpha-deficient mice failed to survive the attenuated serovar Typhimurium infections, suggesting that both endogenous cytokines are essential for regulation of persistent infection with serovar Typhimurium.

scholarly journals Role of YopP in Suppression of Tumor Necrosis Factor Alpha Release by Macrophages during YersiniaInfection

1998 ◽  
Vol 66 (5) ◽  
pp. 1878-1884 ◽  
Author(s):  
Anne Boland ◽  
Guy R. Cornelis
Keyword(s):  
Tumor Necrosis Factor ◽  
Tumor Necrosis Factor Alpha ◽  
Type Strain ◽  
Tumor Necrosis ◽  
Wild Type Strain ◽  
Wild Type ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
Factor Alpha ◽  
Necrosis Factor

ABSTRACT The Yersinia plasmid-encoded Yop virulon enables extracellular adhering bacteria to deliver toxic effector proteins inside their target cells. It includes a type III secretion system (Ysc), at least two translocator proteins (YopB, YopD), and a set of intracellular Yop effectors (YopE, YopH, YopO, YopM, and YopP). Infection of macrophages with a wild-type strain leads to low levels of tumor necrosis factor alpha (TNF-α) release compared to infection with plasmid-cured strains, suggesting that the virulence plasmid encodes a factor impairing the normal TNF-α response of infected macrophages. This effect is correlated with the inhibition of the macrophage mitogen-activated protein kinase (MAPK) activities. To identify the Yop protein responsible for the suppression of TNF-α release, we infected J774A.1 and PU5-1.8 macrophages with a battery of knockout Yersinia enterocolitica mutants and we quantified the TNF-α released. Mutants affected in secretion (yscN), in translocation (yopB and yopD), or in synthesis of all the known Yop effectors (yopH,yopO, yopP, yopE, andyopM polymutants) were unable to block the TNF-α response of the macrophages. In contrast, single yopE,yopH, yopO, and yopM mutants behaved like the wild-type strain. A yopP mutant elicited elevated TNF-α release, and complementation of the yopPmutant or the yop effector polymutant strain withyopP alone led to a drop in TNF-α release. In addition, YopP was also responsible for the inhibition of the extracellular signal-regulated kinase2 (ERK2) and p38 MAPK activities. These results show that YopP is the Yop effector responsible for theYersinia-induced suppression of TNF-α release by infected macrophages.

scholarly journals Changes in polyhydroxyalkanoate granule accumulation make optical density measurement an unreliable method for estimating bacterial growth in Burkholderia thailandensis

2020 ◽  
Vol 66 (3) ◽  
pp. 256-262 ◽  
Author(s):  
Sarah Martinez ◽  
Eric Déziel
Keyword(s):  
Optical Density ◽  
Type Strain ◽  
Wild Type Strain ◽  
Density Measurement ◽  
Viable Cell ◽  
Wild Type ◽  
Burkholderia Thailandensis ◽  
Cell Counts ◽  
Real Cell ◽  
Unreliable Method

Optical density (OD) measurement is the standard method used in microbiology for estimating bacterial concentrations in cultures. However, most studies do not compare these measurements with viable cell counts and assume that they reflect the real cell concentration. Burkholderia thailandensis was recently identified as a polyhydroxyalkanoate (PHA) producer. PHA biosynthesis seems to be coded by an orthologue of the Cupriavidus necator phaC gene. When growing cultures of wild-type strain E264 and an isogenic phaC mutant, we noted a difference in their OD600 values, although viable cell counts indicated similar growth. Investigating the cellular morphologies of both strains, we found that under our conditions the wild-type strain was full of PHA granules, deforming the cells, while the mutant contained no granules. These factors apparently affected the light scattering, making the OD600 values no longer representative of cell density. We show a direct correlation between OD600 values and the accumulation of PHA. We conclude that OD measurement is unreliable for growth evaluation of B. thailandensis because of PHA production. This study also suggests that B. thailandensis could represent an excellent candidate for PHA bioproduction. Correlation between OD measurements and viable cell counts should be verified in any study performed with B. thailandensis.

scholarly journals A genetic analysis of resistance to nystatin inSaccharomyces cerevisiae

1967 ◽  
Vol 9 (2) ◽  
pp. 179-193 ◽  
Author(s):  
K. A. Ahmed ◽  
R. A. Woods
Keyword(s):  
Genetic Analysis ◽  
Resistance Genes ◽  
Type Strain ◽  
Wild Type Strain ◽  
Second Step ◽  
Wild Type ◽  
Recessive Resistance ◽  
Low Concentrations ◽  
Effects Of Temperature ◽  
Resistant Mutants

1. A number of stable nystatin-resistant mutants of the yeastSaccharomyces cerevisiaehave been isolated from platings of a sensitive wild-type strain on low concentrations of the antibiotic.2. These mutants were found to be resistant to 10, 15 or 60 units of drug/ml.3. Analysis of meiotic segregants from crosses of these mutants to wild-type indicate that resistance is determined by two types of genes; resistance genes and modifiers.4. Functional analysis of the mutants demonstrated the existence of three recessive resistance genes,nys-l,nys-2 andnys-3 and thatnys-1 andnys-2 were linked.5. Genetic analysis showed thatnys-1 was affected by two modifiers,Mnys-1 andMnys-2, but that onlyMnys-2 affectednys-2 andnys-3.6. The modifiersMnys-1 andMnys-2 are dominant.7. An investigation of the effects of temperature and medium on resistance demonstrated marked interactions between genotype and environment for both the resistance genes and the modifiers.8. Second-step mutants have been isolated by plating first-step mutants on higher concentrations of the drug. Some of these are resistant to 800 units/ml.9. Some possible mechanisms of nystatin resistance are discussed.

Purification of sex pheromones specific for pMB1 and pMB2 plasmids of Enterococcus faecalis S-48

1995 ◽  
Vol 41 (7) ◽  
pp. 629-632 ◽  
Author(s):  
R. Quirantes ◽  
E. Valdivia ◽  
I. Martín ◽  
M. Martínez-Bueno ◽  
M. Maqueda ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Type Strain ◽  
Sex Pheromones ◽  
Wild Type Strain ◽  
Reversed Phase ◽  
Wild Type ◽  
Pheromone Response ◽  
Conjugative Plasmids ◽  
In The Wild ◽  
Key Words Pheromone

The strain Enterococcus faecalis S-48 carries two large conjugative plasmids (pMB1 and pMB2) encoding for antagonistic substances. The pheromone response of these two plasmids was established by purifying the corresponding sex pheromones, using conventional reversed-phase columns. Plasmid pMB1 responds to pheromone cCF10. Plasmid pMB2 responds to a pheromone with an amino acid sequence identical to that of cPD1 (Phe-Leu-Val-Met-Phe-Leu-Ser-Gly). The two pheromone-responding plasmids coexist in a stable fashion in the wild-type strain E. faecalis S-48.Key words: pheromone response, Enterococcus faecalis, conjugative plasmids.

scholarly journals Cardiac Microlesions Form During Severe Bacteremic Enterococcus faecalis Infection

2020 ◽  
Author(s):  
Armand O Brown ◽  
Kavindra V Singh ◽  
Melissa R Cruz ◽  
Karan Gautam Kaval ◽  
Liezl E Francisco ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Host Response ◽  
Type Strain ◽  
Pneumococcal Disease ◽  
Wild Type Strain ◽  
Protein A ◽  
Virulence Determinants ◽  
Wild Type ◽  
Bond Forming ◽  
Hospital Acquired

Abstract Enterococcus faecalis is a significant cause of hospital-acquired bacteremia. Herein, the discovery is reported that cardiac microlesions form during severe bacteremic E. faecalis infection in mice. The cardiac microlesions were identical in appearance to those formed by Streptococcus pneumoniae during invasive pneumococcal disease. However, E. faecalis does not encode the virulence determinants implicated in pneumococcal microlesion formation. Rather, disulfide bond forming protein A (DsbA) was found to be required for E. faecalis virulence in a Caenorhabditis elegans model and was necessary for efficient cardiac microlesion formation. Furthermore, E. faecalis promoted cardiomyocyte apoptotic and necroptotic cell death at sites of microlesion formation. Additionally, loss of DsbA caused an increase in proinflammatory cytokines, unlike the wild-type strain, which suppressed the immune response. In conclusion, we establish that E. faecalis is capable of forming cardiac microlesions and identify features of both the bacterium and the host response that are mechanistically involved.

scholarly journals Effects of Mutations of RAD50, RAD51, RAD52, and Related Genes on Illegitimate Recombination in Saccharomyces cerevisiae

Genetics ◽  
1996 ◽  
Vol 142 (2) ◽  
pp. 383-391 ◽  
Author(s):  
Yasumasa Tsukamoto ◽  
Jun-ichi Kato ◽  
Hideo Ikeda
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Quantitative Analysis ◽  
Structural Analysis ◽  
Recombination Rate ◽  
Type Strain ◽  
Negative Selection ◽  
Wild Type Strain ◽  
Illegitimate Recombination ◽  
Wild Type ◽  
In The Wild

Abstract To examine the mechanism of illegitimate recombination in Saccharomyces cerevisiae, we have developed a plasmid system for quantitative analysis of deletion formation. A can1 cyh2 cell carrying two negative selection markers, the CAN1 and CYH2 genes, on a YCp plasmid is sensitive to canavanine and cycloheximide, but the cell becomes resistant to both drugs when the plasmid has a deletion over the CAN1 and CYH2 genes. Structural analysis of the recombinant plasmids obtained from the resistant cells showed that the plasmids had deletions at various sites of the CAN1-CYH2 region and there were only short regions of homology (1-5 bp) at the recombination junctions. The results indicated that the deletion detected in this system were formed by illegitimate recombination. Study on the effect of several rad mutations showed that the recombination rate was reduced by 30-, 10-, 10-, and 10-fold in the rad52, rad50, mre11, and xrs2 mutants, respectively, while in the rud51, 54, 55, and 57 mutants, the rate was comparable to that in the wild-type strain. The rad52 mutation did not affect length of homology at junction sites of illegitimate recombination.

scholarly journals The role of Zur-regulated lipoprotein A in bacterial morphology, antimicrobial susceptibility, and production of outer membrane vesicles in Acinetobacter baumannii

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Nayeong Kim ◽  
Hyo Jeong Kim ◽  
Man Hwan Oh ◽  
Se Yeon Kim ◽  
Mi Hyun Kim ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Outer Membrane ◽  
Mutant Strain ◽  
Antimicrobial Susceptibility ◽  
Type Strain ◽  
Wild Type Strain ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Wild Type ◽  
Bacterial Morphology

Abstract Background Zinc uptake-regulator (Zur)-regulated lipoprotein A (ZrlA) plays a role in bacterial fitness and overcoming antimicrobial exposure in Acinetobacter baumannii. This study further characterized the zrlA gene and its encoded protein and investigated the roles of the zrlA gene in bacterial morphology, antimicrobial susceptibility, and production of outer membrane vesicles (OMVs) in A. baumannii ATCC 17978. Results In silico and polymerase chain reaction analyses showed that the zrlA gene was conserved among A. baumannii strains with 97–100% sequence homology. Recombinant ZrlA protein exhibited a specific enzymatic activity of D-alanine-D-alanine carboxypeptidase. Wild-type A. baumannii exhibited more morphological heterogeneity than a ΔzrlA mutant strain during stationary phase. The ΔzrlA mutant strain was more susceptible to gentamicin than the wild-type strain. Sizes and protein profiles of OMVs were similar between the wild-type and ΔzrlA mutant strains, but the ΔzrlA mutant strain produced 9.7 times more OMV particles than the wild-type strain. OMVs from the ΔzrlA mutant were more cytotoxic in cultured epithelial cells than OMVs from the wild-type strain. Conclusions The present study demonstrated that A. baumannii ZrlA contributes to bacterial morphogenesis and antimicrobial resistance, but its deletion increases OMV production and OMV-mediated host cell cytotoxicity.

scholarly journals FixJ family regulator AcfR of Azorhizobium caulinodans is involved in symbiosis with the host plant

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Wei Liu ◽  
Xue Bai ◽  
Yan Li ◽  
Haikun Zhang ◽  
Xiaoke Hu
Keyword(s):  
Signal Transduction ◽  
Host Plant ◽  
Type Strain ◽  
Wild Type Strain ◽  
Response Regulator ◽  
Azorhizobium Caulinodans ◽  
Wild Type ◽  
Response Regulators ◽  
Free Living ◽  
Two Component

Abstract Background A wide variety of bacterial adaptative responses to environmental conditions are mediated by signal transduction pathways. Two-component signal transduction systems are one of the predominant means used by bacteria to sense the signals of the host plant and adjust their interaction behaviour. A total of seven open reading frames have been identified as putative two-component response regulators in the gram-negative nitrogen-fixing bacteria Azorhizobium caulinodans ORS571. However, the biological functions of these response regulators in the symbiotic interactions between A. caulinodans ORS571 and the host plant Sesbania rostrata have not been elucidated to date. Results In this study, we identified and investigated a two-component response regulator, AcfR, with a phosphorylatable N-terminal REC (receiver) domain and a C-terminal HTH (helix-turn-helix) LuxR DNA-binding domain in A. caulinodans ORS571. Phylogenetic analysis showed that AcfR possessed close evolutionary relationships with NarL/FixJ family regulators. In addition, six histidine kinases containing HATPase_c and HisKA domains were predicted to interact with AcfR. Furthermore, the biological function of AcfR in free-living and symbiotic conditions was elucidated by comparing the wild-type strain and the ΔacfR mutant strain. In the free-living state, the cell motility behaviour and exopolysaccharide production of the ΔacfR mutant were significantly reduced compared to those of the wild-type strain. In the symbiotic state, the ΔacfR mutant showed a competitive nodule defect on the stems and roots of the host plant, suggesting that AcfR can provide A. caulinodans with an effective competitive ability for symbiotic nodulation. Conclusions Our results showed that AcfR, as a response regulator, regulates numerous phenotypes of A. caulinodans under the free-living conditions and in symbiosis with the host plant. The results of this study help to elucidate the involvement of a REC + HTH_LuxR two-component response regulator in the Rhizobium-host plant interaction.

scholarly journals Mab_3083c Is a Homologue of RNase J and Plays a Role in Colony Morphotype, Aggregation, and Sliding Motility of Mycobacterium abscessus

2021 ◽  
Vol 9 (4) ◽  
pp. 676
Author(s):  
Ting-Yu Liu ◽  
Sheng-Hui Tsai ◽  
Jenn-Wei Chen ◽  
Yu-Ching Wang ◽  
Shiau-Ting Hu ◽  
...  
Keyword(s):  
Type Strain ◽  
Wild Type Strain ◽  
Opportunistic Pathogen ◽  
Mycobacterium Abscessus ◽  
Colony Morphology ◽  
Clinical Strain ◽  
Immunocompromised Patients ◽  
Clinical Infection ◽  
Wild Type ◽  
Sliding Motility

Mycobacterium abscessus is an opportunistic pathogen causing human diseases, especially in immunocompromised patients. M. abscessus strains with a rough morphotype are more virulent than those with a smooth morphotype. Morphotype switch may occur during a clinical infection. To investigate the genes involved in colony morphotype switching, we performed transposon mutagenesis in a rough clinical strain of M. abscessus. A morphotype switching mutant (smooth) named mab_3083c::Tn was obtained. This mutant was found to have a lower aggregative ability and a higher sliding motility than the wild type strain. However, its glycopeptidolipid (GPL) content remained the same as those of the wild type. Complementation of the mutant with a functional mab_3083c gene reverted its morphotype back to rough, indicating that mab_3083c is associated with colony morphology of M. abscessus. Bioinformatic analyses showed that mab_3083c has a 75.4% identity in amino acid sequence with the well-characterized ribonuclease J (RNase J) of M. smegmatis (RNase JMsmeg). Complementation of the mutant with the RNase J gene of M. smegmatis also switched its colony morphology from smooth back to rough. These results suggest that Mab_3083c is a homologue of RNase J and involved in regulating M. abscessus colony morphotype switching.

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