Genes Encoding Bacteriocins and Their Expression and Potential Virulence Factors of Enterococci Isolated from Wood Pigeons (Columba palumbus)

2006 ◽  
Vol 69 (3) ◽  
pp. 520-531 ◽  
Author(s):  
MARÍA MARTÍN ◽  
JORGE GUTIÉRREZ ◽  
RAQUEL CRIADO ◽  
CARMEN HERRANZ ◽  
LUIS M. CINTAS ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Virulence Factors ◽  
Virulence Genes ◽  
Pcr Amplification ◽  
Virulence Gene ◽  
Antagonistic Activity ◽  
Mass Spectrometry Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virulence Determinants ◽  
Spectrometry Analysis

Samples of the intestinal content and carcasses of wood pigeons (Columba palumbus) were evaluated for enterococci with antimicrobial activity. Enterococcus faecium comprised the largest enterococcal species with antagonistic activity, followed by Enterococcus faecalis and Enterococcus columbae. PCR amplification of genes coding bacteriocins and determination of their nucleotide sequence, and the use of specific antipeptide bacteriocin antibodies and a noncompetitive indirect enzyme-linked immunosorbent assay, permitted characterization of enterococci coding that described bacteriocins and their expression. The efaAfm determinant was the only virulence gene detected in E. faecium, whereas E. faecalis showed a larger number of virulence determinants, and E. columbae did not carry any of the virulence genes examined. Although all E. faecalis isolates manifested a potent direct antimicrobial activity, no activity was detected in supernatants of producer cells. Purification of the antagonistic activity of E. columbae PLCH2 showed multiple chromatographic fragments after matrix-assisted laser desorption–ionization time-of-flight mass spectrometry analysis, suggesting the active peptide(s) had not yet purified to homogeneity. Bacteriocinogenic E. faecium and E. columbae isolates may be considered hygienic for production of enterocins and potentially safe due to their low incidence of potential virulence genes and susceptibility of most relevant clinical antibiotics. However, the presence among the enterococci of E. faecalis strains with a potent antagonistic activity and multiple virulence factors is an issue that must be considered further.

scholarly journals Ibuprofen-mediated potential inhibition of biofilm development and quorum sensing inPseudomonas aeruginosa

2019 ◽  
Author(s):  
Lu Dai ◽  
Tian-qi Wu ◽  
Zhong-hui Wang ◽  
Ruoyu Yuan ◽  
Ye Ding ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Quorum Sensing ◽  
Virulence Factors ◽  
Mass Spectrometry Analysis ◽  
Significant Suppression ◽  
Spectrometry Analysis ◽  
Acridine Orange Staining ◽  
Drug Concentrations ◽  
Associated Proteins ◽  
Biofilm Development

AbstractPseudomonas aeruginosais one of the leading causes of opportunistic and hospital-acquired infections worldwide. The infection withP. aeruginosais frequently linked with clinical treatment difficulties given drug resistance and abuse of antibiotics. Ibuprofen, a widely used non-steroidal anti-inflammatory drug, has been previously reported to exert antimicrobial activity, although the specific mechanism of its action requires additional investigation. Given the regulation effects on quorum sensing (QS), we hypothesized that inhibition ofP. aeruginosawith ibuprofen is linked with the QS systems. First, we assessed the action of ibuprofen inP. aeruginosaby measuring CFU. The antimicrobial activity of ibuprofen was evaluated by crystal violent staining and acridine orange staining at various drug concentrations (0, 50, 75, and 100 μg/mL). Moreover, the effect of ibuprofen on different QS virulence factors, such as pyocyanin, elastase, protease, and rhamnolipids, was assessed revealing a concentration-dependent decrease (P<0.05). The effect of ibuprofen was confirmed by liquid chromatography/mass spectrometry analysis of 3-oxo-C12-HSL and C4-HSL production. In addition, qRT-PCR results identified significant suppression of Las and Rhl gene expression after 18 hours of treatment with ibuprofen (P<0.05), with the most significant suppression observed at the concentration of 75 μg/mL. Functional complementation with exogenous 3-oxo-C12-HSL and C4-HSL suggested that C4-HSL can recover the production of virulence factors and biofilm formation inP. aeruginosa.Molecular docking of ibuprofen with QS-associated proteins revealed high binding affinity. In summary, the results suggest that ibuprofen is a candidate drug for the treatment of clinical infections withP. aeruginosa.

scholarly journals Enterococcus faecium isolated from healthy dogs for potential use as probiotics

2020 ◽  
Vol 23 (2) ◽  
pp. 197-205
Author(s):  
K. A. Abd El-Razik ◽  
E. S. Ibrahim ◽  
A. M. Younes ◽  
A. A. Arafa ◽  
A. S. M. Abuelnaga ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Molecular Identification ◽  
Enterococcus Faecium ◽  
Virulence Genes ◽  
Pcr Amplification ◽  
Virulence Determinants ◽  
Specific Pcr ◽  
Healthy Dogs ◽  
Potential Use

This study aimed to isolate and identify enterococci obtained from fresh faecal swabs of 16 healthy dogs. Following molecular identification, all isolates were screened against the most critical virulence factors as well as enterocin (bacteriocin) determinants to confirm that the isolated enterococcus was safe to be used as host-specific probiotic. Enterococcus faecium was isolated and confirmed in 8 out of the 16 samples. Regarding the assessment of the virulence determinants, E. faecium strains were negative for tested (gelE and esp) virulence genes. Furthermore, the genome was evaluated for the incidence of five known enterocin genes by specific PCR amplification. Four strains encoding entAS-48 gene were found, while only one strain harboured the entL50A/B gene. Based on these results, five of the E. faecium isolated in this study were considered as promising probiotic candidates for dogs.

Virulence determinants and production of extracellular enzymes in Enterococcus spp. from surface water sources

2016 ◽  
Vol 73 (8) ◽  
pp. 1817-1824 ◽  
Author(s):  
Lesego Gertrude Molale ◽  
Cornelius Carlos Bezuidenhout
Keyword(s):  
Surface Water ◽  
Virulence Factors ◽  
Virulence Genes ◽  
Extracellular Enzymes ◽  
Virulence Gene ◽  
Water Sources ◽  
Virulence Determinants ◽  
North West ◽  
Enterococcus Spp

Virulence factors in Enterococcus may be indicative of potential pathogenicity. The aim of this study was to determine the relationship between the presence of clinically relevant virulence genes, in Enterococcus spp. from environmental water, and their in vitro expression. One hundred and twenty-four Enterococcus isolates (seven species), from five surface water systems in the North West Province, South Africa, were screened for the presence of asa1, cylA, esp, gelE and hyl using polymerase chain reaction. The expression of cylA, hyl and gelE was determined by phenotypic assessments. Sixty-five percent of the isolates were positive for one virulence gene and 13% for two or more. Most frequently detected genes were gelE (32%) and cylA (28%). Enterococcal surface protein was absent in all isolates screened. The presence of virulence genes was correlated with their extracellular enzyme production. The results show that a large percentage of these environmental Enterococcus spp. possess virulence factors that could be expressed in vitro. This is a cause for concern and could have implications for individuals using this water for recreational and cultural purposes. Further investigation is required into the sources of these potential pathogenic Enterococcus isolates and measures to minimize their presence in water sources.

Epidemiological features of nosocomial Klebsiella pneumoniae: virulence and resistance determinants

2021 ◽  
Author(s):  
Mai M Zafer ◽  
Maha M El Bastawisie ◽  
Mona Wassef ◽  
Amira FA Hussein ◽  
Mohammed A Ramadan
Keyword(s):  
Klebsiella Pneumoniae ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Pcr Amplification ◽  
Virulence Gene ◽  
Healthcare Setting ◽  
Virulence Determinants ◽  
Tertiary Healthcare ◽  
Insight Into ◽  
Common Sequence

Aim: The authors aimed to examine antibiotic resistance genes and representative virulence determinants among 100 Klebsiella pneumoniae isolates with an emphasis on capsular serotypes and clonality of some of the isolates. Methods: PCR amplification of ( rmpA, rmpA2, iutA, iroN and IncHI1B plasmid) and (NDM, OXA-48, KPC, CTX-M-15, VIM, IMP, SPM) was conducted. Wzi sequencing and multilocus sequence typing (MLST) were performed. Results: K2 was the only detected serotype in the authors' collection. RMPA2 was the most common capsule-associated virulence gene detected. All studied isolates harbored OXA-48-like (100%) and NDM (43%) (n = 43). ST147 was the most common sequence type. Conclusion: This work provides insight into the evolution of the coexistence of virulence and resistance genes in a tertiary healthcare setting in Cairo, Egypt.

scholarly journals GAS CHROMATOGRAPHY-MASS SPECTROMETRY ANALYSIS AND IN VITRO ANTIMICROBIAL SCREENING OF WEDELIA GLAUCA (ORTEGA) O. HOFFM. EX HICKEN

2017 ◽  
Vol 10 (11) ◽  
pp. 109
Author(s):  
Krishnavignesh L Krishnavignesh ◽  
Mahalakshmipriya A ◽  
Ramesh M
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Antimicrobial Activity ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Diffusion Method ◽  
Spectrometry Analysis ◽  
Chromatography Mass Spectrometry ◽  
Ms Analysis

  Objective: Continued resistance toward the antibiotics urges us to explore newer antibiotics. Plants are being the safer source of antibiotics with lesser or no side effects. This study was designed to study the presence of phytochemical constituents and antibacterial activity of leaf and flower extracts of Wedelia glauca against urinary tract infection causing pathogens.Methods: The plant leaves were extracted with five different solvents based on the polarity. The extraction was done using soxhalation. Antimicrobial activity was determined by agar well diffusion method for both the sample and standard. The acetone plant extract was subjected to gas chromatography-mass spectrometry (GC-MS) analysis for screening phytoconstituents.Results: Preliminary phytochemical screening revealed the presence of diverse phytoconstituents in the plant. The different extracts exhibited a considerable antimicrobial potential. Among the solvents used acetone extract showed comparably better antimicrobial activity with 100% of inhibition rate with the maximum zone of inhibition of 1.6±0.77 mm against Staphylococcus sp. and Aspergillus sp. at the concentration of 5 mg. GC-MS analysis provided 8 major peaks which revealed the existence of a variety of bioactive compounds which may attribute to the efficacy of the plant.Conclusion: W. glauca leaf and flower extracts displayed a broad spectrum of antibacterial and antifungal activity and can be considered as a potential source of newer antibiotic compounds.

Virulence Gene Pattern of Pasteurella multocida Isolates of Buffalo in Association to Capsule Biosynthesis Genes

2020 ◽  
Author(s):  
N. Sujatha ◽  
K. Lakshmi Kavitha ◽  
K.V. Subramanyam ◽  
T. Srinivasa Rao ◽  
R.N. Ramani Pushpa
Keyword(s):  
Virulence Factors ◽  
Pasteurella Multocida ◽  
Virulence Genes ◽  
Systemic Disease ◽  
Virulence Gene ◽  
Disease Status ◽  
Host Cells ◽  
Wide Range ◽  
Serotype B ◽  
Apparently Healthy

Background: Pasteurella multocida is the causative agent of many economically important diseases in a wide range of hosts. The mechanisms by which these bacteria can invade the mucosa, evade innate immunity and cause systemic disease are slowly being elucidated. Many key virulence factors are yet to be identified, including those required for initial attachment and invasion of host cells and for persistence in a relatively nutrient poor and hostile environment. This has led to intensive research to understand host adaptation mechanisms and virulence factors in order to develop effective vaccines. Methods: The present study was carried out to know the distribution of virulence genes viz., haemoglobin binding proteins (hgbA and hgbB), outer membrane protein (ompH), fimbrial antigen (ptfA), filamentous haemagglutinin (pfhA) and transferrin binding protein (tbpA) by PCR in P. multocida CapA isolates from apparently healthy or carrier animals and CapB isolates from field Haemorrhagic septicemia (HS) cases to monitor the epidemiological associations of virulence genes in Cap A and Cap B isolates.Result: The study revealed that all the six virulence associated genes were present in Cap B isolates. None of the Cap A isolates harboured tbpA and pfhA genes. These two genes were closely related to serotype B causing Haemorrhagic septicemia and were epidemiologically associated with disease status.

scholarly journals Molecular Characterization of Shiga Toxin-Producing Escherichia coli Strains Isolated in Poland

2016 ◽  
Vol 65 (3) ◽  
pp. 261-269 ◽  
Author(s):  
Aleksandra Januszkiewicz ◽  
Waldemar Rastawicki
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Shiga Toxin ◽  
Virulence Gene ◽  
Virulence Determinants ◽  
E Coli ◽  
Heat Stable ◽  
Food Borne ◽  
Wide Range ◽  
Uremic Syndrome

Shiga toxin-producing Escherichia coli (STEC) strains also called verotoxin-producing E. coli (VTEC) represent one of the most important groups of food-borne pathogens that can cause several human diseases such as hemorrhagic colitis (HC) and hemolytic – uremic syndrome (HUS) worldwide. The ability of STEC strains to cause disease is associated with the presence of wide range of identified and putative virulence factors including those encoding Shiga toxin. In this study, we examined the distribution of various virulence determinants among STEC strains isolated in Poland from different sources. A total of 71 Shiga toxin-producing E. coli strains isolated from human, cattle and food over the years 1996 – 2010 were characterized by microarray and PCR detection of virulence genes. As stx1a subtype was present in all of the tested Shiga toxin 1 producing E. coli strains, a greater diversity of subtypes was found in the gene stx2, which occurred in five subtypes: stx2a, stx2b, stx2c, stx2d, stx2g. Among STEC O157 strains we observed conserved core set of 14 virulence factors, stable in bacteria genome at long intervals of time. There was one cattle STEC isolate which possessed verotoxin gene as well as sta1 gene encoded heat-stable enterotoxin STIa characteristic for enterotoxigenic E. coli. To the best of our knowledge, this is the first comprehensive analysis of virulence gene profiles identified in STEC strains isolated from human, cattle and food in Poland. The results obtained using microarrays technology confirmed high effectiveness of this method in determining STEC virulotypes which provides data suitable for molecular risk assessment of the potential virulence of this bacteria.

scholarly journals Analysis of virulence factors and antibiotic resistance genes in Group B Streptococcus from clinical samples

2020 ◽  
Author(s):  
Raymond Mudzana ◽  
Rooyen T Mavenyengwa ◽  
Muchaneta Gudza-Mugabe
Keyword(s):  
Antibiotic Resistance ◽  
Pregnant Women ◽  
Virulence Factors ◽  
Resistance Genes ◽  
Virulence Genes ◽  
Group B Streptococcus ◽  
Antibiotic Resistance Genes ◽  
Virulence Gene ◽  
Clinical Samples ◽  
Group B

Abstract Background: Streptococcus agalacticae is one of the most important causative agents of serious infections among neonates. Group B Streptococcus (GBS) virulence factors are important in the development of vaccines, whilst antibiotic resistance genes are necessary in understanding the resistance mechanisms used by these pathogens. This study was carried out to identify the virulence genes and antibiotic resistance genes associated with GBS isolated from pregnant women.Methods: A total of 43 GBS isolates were obtained from vaginal samples that were collected from all HIV positive and HIV negative women who were 13-35 weeks pregnant attending Antenatal Care at both Chitungwiza and Harare Central Hospitals in Zimbabwe. Identification tests of GBS isolates was done using standard bacteriological methods including molecular tests. Antibiotic susceptibility testing using 3 antibiotics was done using the modified Kirby-Bauer method. The boiling method was used to extract DNA and Polymerase Chain Reaction (PCR) was used to screen for 13 genes in the isolates. Data was fed into SPSS 24.0 and the Spearman rank correlation test used to determine any correlation among genes.Results: Nine distinct virulence gene profiles were identified. The profiles hly-scpB-bca-rib 37.2% (16/43) and hly-scpB-bca 18.6% (8/43) were common among GBS isolates. The following virulence gene frequencies were obtained namely hly 97.8% (42/43), scpB 90.1% (39/43), bca 86.0% (37/43), rib 69.8% (30/43) and bac 11.6% (5/43). Antibiotic resistance genes showed high frequencies for tetM 97.6% (41/42) and low frequencies for ermB 34.5% (10/29), ermTR 10.3% (3/29), mefA 3.4% (1/29), tetO 2.4% (1/42) and linB 0% (0/35). The atr housekeeping gene amplification yielded 100% (43/43) positive results, whilst the mobile genetic element IS1548 yielded a low 9.3% (4/43).Conclusion: The study showed a high prevalence of multiple virulence genes hly, scpB, bca and rib in S. agalactiae strains isolated from pregnant women. Tetracycline resistance was found to be predominantly caused by the tetM gene, whilst macrolide resistance was predominantly due to the presence of erm methylase, with the ermB gene being more prevalent. It was also observed that in vitro phenotypic resistance is not always accurately predicted by resistance genotypes.

scholarly journals Virulence and antibiotic resistance profile of avian Escherichia coli strains isolated from colibacillosis lesions in central of Algeria

2019 ◽  
Vol 12 (11) ◽  
pp. 1840-1848 ◽  
Author(s):  
Nacima Meguenni ◽  
Nathalie Chanteloup ◽  
Angelina Tourtereau ◽  
Chafika Ali Ahmed ◽  
Saliha Bounar-Kechih ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Virulence Factors ◽  
Virulence Genes ◽  
Iron Acquisition ◽  
Virulence Gene ◽  
Resistance Profile ◽  
E Coli ◽  
Antibiotic Resistance Profile ◽  
Embryo Lethality

Background and Aim: Avian pathogenic Escherichia coli cause extensive mortality in poultry flocks, leading to extensive economic losses. To date, in Algeria, little information has been available on virulence potential and antibiotics resistance of avian E. coli isolates. Therefore, the aim of this study was the characterization of virulence genes and antibiotic resistance profile of Algerian E. coli strains isolated from diseased broilers. Materials and Methods: In this study, 43 avian E. coli strains isolated from chicken colibacillosis lesions at different years were analyzed to determine their contents in 10 virulence factors by polymerase chain reaction, antimicrobial susceptibility to 22 antibiotics belonging to six different chemical classes and genomic diversity by pulsed-field gel electrophoresis (PFGE). Results: Mainly E. coli isolates (58.1%) carried two at six virulence genes and the most frequent virulence gene association detected were ompT (protectin), hlyF (hemolysin) with 55.8% (p<0.001), and iroN, sitA (iron acquisition/uptake systems), and iss (protectin) with 41.8% (p<0.001). Some strains were diagnosed as virulent according to their virulence gene profile. Indeed, 23.25% of the isolates harbored iroN, ompT, hlyF, iss, and sitA combination, 14% ompT, hlyF, and frzorf4 (sugar metabolism), and 11,6% iroN, hlyF, ompT, iss, iutA (iron acquisition/uptake systems), and frzorf4. The chicken embryo lethality assay performed on five isolates confirmed the potential virulence of these strains. All isolates submitted to PFGE analysis yielded different genetic profiles, which revealed their diversity. Overall, 97.2% of the isolates were resistant to at least one antibiotic and 53.5% demonstrated multi-antimicrobial resistance to three different antimicrobial classes. The highest resistance levels were against nalidixic acid (83.4%), amoxicillin and ampicillin (83.3%), ticarcillin (80.5%), pipemidic acid (75%), and triméthoprim-sulfamethoxazole (66.6%). For beta-lactam class, the main phenotype observed belonged to broad-spectrum beta-lactamases. However, extended-spectrum beta-lactamase associated with three at six virulence factors was also detected in 13 isolates. Two of them were attested virulent as demonstrated in the embryo lethality test which constitutes a real public threat. Conclusion: It would be imperative in avian production to discourage misuse while maintaining constant vigilance guidelines and regulations, to limit and rationalize antimicrobial use.

scholarly journals Identification of Extracellular N-Acylhomoserine Lactone Acylase from a Streptomyces sp. and Its Application to Quorum Quenching

2005 ◽  
Vol 71 (5) ◽  
pp. 2632-2641 ◽  
Author(s):  
Sun-Yang Park ◽  
Hye-Ok Kang ◽  
Hak-Sun Jang ◽  
Jung-Kee Lee ◽  
Bon-Tag Koo ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Pathogenic Bacteria ◽  
Quorum Quenching ◽  
Homoserine Lactone ◽  
Amino Acid Sequences ◽  
Amide Bond ◽  
Mass Spectrometry Analysis ◽  
Penicillin G ◽  
Spectrometry Analysis ◽  
Acyl Chains

ABSTRACT N-Acylhomoserine lactones (AHLs) play an important role in regulating virulence factors in pathogenic bacteria. Recently, the enzymatic inactivation of AHLs, which can be used as antibacterial targets, has been identified in several soil bacteria. In this study, strain M664, identified as a Streptomyces sp., was found to secrete an AHL-degrading enzyme into a culture medium. The ahlM gene for AHL degradation from Streptomyces sp. strain M664 was cloned, expressed heterologously in Streptomyces lividans, and purified. The enzyme was found to be a heterodimeric protein with subunits of approximately 60 kDa and 23 kDa. A comparison of AhlM with known AHL-acylases, Ralstonia strain XJ12B AiiD and Pseudomonas aeruginosa PAO1 PvdQ, revealed 35% and 32% identities in the deduced amino acid sequences, respectively. However, AhlM was most similar to the cyclic lipopeptide acylase from Streptomyces sp. strain FERM BP-5809, exhibiting 93% identity. A mass spectrometry analysis demonstrated that AhlM hydrolyzed the amide bond of AHL, releasing homoserine lactone. AhlM exhibited a higher deacylation activity toward AHLs with long acyl chains rather than short acyl chains. Interestingly, AhlM was also found to be capable of degrading penicillin G by deacylation, showing that AhlM has a broad substrate specificity. The addition of AhlM to the growth medium reduced the accumulation of AHLs and decreased the production of virulence factors, including elastase, total protease, and LasA, in P. aeruginosa. Accordingly, these results suggest that AHL-acylase, AhlM could be effectively applied to the control of AHL-mediated pathogenicity.

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