scholarly journals Identification of Ciprofloxacin-ResistantCampylobacter jejuni by Use of a Fluorogenic PCR Assay

2000 ◽  
Vol 38 (11) ◽  
pp. 3971-3978 ◽  
Author(s):  
David L. Wilson ◽  
Sheila R. Abner ◽  
Thomas C. Newman ◽  
Linda S. Mansfield ◽  
John E. Linz
Keyword(s):  
Antimicrobial Agents ◽  
Sequence Data ◽  
Dna Gyrase ◽  
Species Level ◽  
Primary Target ◽  
Pcr Assay ◽  
Nucleotide Sequence Data ◽  
Rapid Pcr ◽  
Fluoroquinolone Antibiotics ◽  
Ciprofloxacin Resistance

Fluoroquinolones are one class of antimicrobial agents commonly used to treat severe Campylobacter jejuni infection.C. jejuni strains resistant to high levels of the fluoroquinolone ciprofloxacin (MIC ≥16 μg/ml) have been predominantly characterized with a C→T transition in codon 86 ofgyrA. The gyrA gene encodes one subunit of DNA gyrase, which is a primary target for fluoroquinolone antibiotics. This study establishes a rapid PCR-based TaqMan method for identifying ciprofloxacin-resistant C. jejuni strains that carry the C→T transition in codon 86 of gyrA. The assay uses real-time detection, eliminating the need for gel electrophoresis. Optimization of the assay parameters using purified Campylobacter DNA resulted in the ability to detect femtogram levels of DNA. The method should be useful for monitoring the development of ciprofloxacin resistance in C. jejuni. Compiled nucleotide sequence data on the quinolone resistance-determining region of gyrA inCampylobacter indicate that sequence comparison of this region is a useful method for tentative identification ofCampylobacter isolates at the species level.

scholarly journals Identification of DNA gyrase Subunit a Mutations Associated with Ciprofloxacin Resistance in Staphylococcus aureus Isolated from Nasal Infection in Kurdistan-Iran

2017 ◽  
Vol 7 (1) ◽  
pp. 186
Author(s):  
Roya Darbani ◽  
Chiako Farshadfar ◽  
Somayeh Tavana ◽  
Hamidreza Saljoughi ◽  
Sheida Sadat Zonouri
Keyword(s):  
Staphylococcus Aureus ◽  
Free Energy ◽  
Molecular Docking ◽  
Dna Gyrase ◽  
Amino Acid Sequences ◽  
Nucleotide Sequencing ◽  
Molegro Virtual Docker ◽  
Fluoroquinolone Antibiotics ◽  
Ciprofloxacin Resistance ◽  
Free Energy Of Binding

Fluoroquinolone antibiotics such as ciprofloxacin are useful drugs against infections caused by Staphylococcus aureus and mutations in DNA gyrase which control bacterial DNA topology, can be one of the reason of occurrence resistance to this class of antibiotics. Therefore finding new mutations and study of the quinolone interaction with mutated GyrA can provide important issues for explanation resistance. In this study 5 ciprofloxacin resistance Staphylococcus aureus isolated among 50 collected S.aureus strains. By PCR testing, gyrA genes in resistance strains was amplified and nucleotide sequencing was done. Nucleotide sequences translate to amino acid sequences then by blastp homology between each GyrA mutant and reference GyrA were compared and mutations were recognized, at last molecular docking were done for GyrA protein and ciprofloxacin, based on free energy of binding decide if the mutations are responsible of resistance or not. The results show glutamic acid and threonine adjacent to each other in common positions 21-22, 32-33, 65-66, 84-85, 101-102, 106-107, 128-129 and 138-139 in all 5 strains were inserted . In order to finding association between mutations and ciprofloxacin resistance molecular docking by Molegro Virtual Docker 5.5 was done. Free energy of binding between reference GyrA- ciprofloxacin and mutant GyrA- ciprofloxacin were -92.3477 and -73.1642 respectively. We conclude different mutations can be affected structure of GyrA and make ciprofloxacin resistance. Finding these kinds of mutations are important and preventing them is indispensable.

scholarly journals Molecular identification and antimicrobial activity of endophytic fungi isolated from Heritiera fomes (Buch. -Ham), a mangrove plant of the Sundarbans

2020 ◽  
Vol 9 (1) ◽  
Author(s):  
Tauhidur Rahman Nurunnabi ◽  
Sabiha Sarwar ◽  
Farah Sabrin ◽  
Farzana Alam ◽  
Lutfun Nahar ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Endophytic Fungi ◽  
Antimicrobial Agents ◽  
Sequence Data ◽  
Micrococcus Luteus ◽  
Mangrove Plant ◽  
Nucleotide Sequence Data ◽  
Methanolic Extracts ◽  
Heritiera Fomes ◽  
The Sundarbans

Abstract Background Endophytic fungi play a vital role in plant defense system by secreting various antimicrobial agents. To evaluate the antimicrobial activity of the endophytic fungi of the mangrove plant Heritiera fomes (Buch. -Ham), plants were collected from the Sundarbans, Bangladesh. The endophytic fungi were subsequently isolated and identified. Results Fifty-five endophytic fungi were isolated from the leaves, root, and bark of H. fomes (Buch. -Ham). Genomic DNA was extracted for PCR (polymerase chain reaction) by specific primers to multiply ITS region and sequences were determined. Nucleotide sequence data were submitted to the Genbank and accession number for each fungal strain was obtained. Antimicrobial activity of the ethyl acetate (EtOAc) and methanolic extracts of eleven species from both fermentation and mycelium, respectively, were analyzed by microtiter plate-based antimicrobial assay incorporating resazurin as an indicator of cell growth against two Gram-positive bacteria namely Staphylococcus aureus NCTC 12981 and Micrococcus luteus NCTC 7508, two Gram-negative bacteria namely Escherichia coli NCTC 12241 and Pseudomonas aeruginosa NCTC 12903, and a fungus Candida albicans ATCC 90028. All the endophytic fungal extracts exhibited antimicrobial activities against more than one-tested pathogenic microbial strains. Overall, methanolic extracts showed greater activity than EtOAc extracts. Pseudopestalotiopsis camelliae-sinensis, Pestalotiopsis microspora, and Penicillium copticola were the most active endophytic fungal strains and exhibited strong inhibitory activity against the microorganisms under investigation and their MIC values ranged from 0.0024 to 5.0 mg/mL. Methanolic extracts of both P. camelliae-sinensis and P. microspora showed the highest antibacterial activity (MIC value of 0.0024 mg/mL) against P. aeruginosa NCTC 12903. Conclusion This study showed that the isolated and identified endophytic fungi from H. fomes (Buch. -Ham) could be potential sources of antimicrobial agents.

scholarly journals Zinc can counteract selection for ciprofloxacin resistance

2019 ◽  
Author(s):  
Michiel Vos ◽  
Louise Sibleyras ◽  
Lai Ka Lo ◽  
Elze Hesse ◽  
William Gaze ◽  
...  
Keyword(s):  
Bacterial Resistance ◽  
Antimicrobial Agents ◽  
E Coli ◽  
Resistance Evolution ◽  
Synergistic Interactions ◽  
Selection Dynamics ◽  
Low Concentrations ◽  
Fluoroquinolone Antibiotics ◽  
Ciprofloxacin Resistance ◽  
Zinc Cations

AbstractAntimicrobial resistance (AMR) has emerged as one of the most pressing global threats to public health. AMR evolution occurs in the clinic but also in the environment, where low concentrations of antibiotics and heavy metals can respectively select and co-select for resistance. While the selective potential for AMR of both antibiotics and metals is increasingly well-characterized, studies exploring the combined effect of both types of selective agents are rare. It has previously been demonstrated that fluoroquinolone antibiotics such as ciprofloxacin can chelate metal ions. To investigate how ciprofloxacin resistance is affected by the presence of metals, we quantified selection dynamics between a ciprofloxacin-susceptible and an isogenic ciprofloxacin-resistant Escherichia coli MG1655 strain across a gradient of ciprofloxacin concentrations in the presence and absence of Zinc cations (Zn2+). The minimal selective concentration (MSC) for ciprofloxacin resistance significantly increased up to 5-fold in the presence of Zn2+. No such effect on the MSC was found for gentamicin, an antibiotic not known to chelate zinc cations. Environmental pollution usually consists of complex mixtures of antimicrobial agents. Our findings highlight the importance of taking antagonistic as well as additive or synergistic interactions between different chemical compounds into account when considering their effect on bacterial resistance evolution.Graphical abstractOne sentence summaryThe minimal selective concentration for a ciprofloxacin resistant E. coli strain increases up to 5-fold in the presence of Zinc cations.

scholarly journals PCR-based Specific Detection of Bacillus in Liquid Organic Fertilizer

2019 ◽  
Vol 1 (1) ◽  
pp. 44-47
Author(s):  
Iman Hidayat ◽  
Nur Laili ◽  
Dwi Agustiyani ◽  
Sarjiya Antonius
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Sequence Data ◽  
Organic Fertilizer ◽  
Bacillus Species ◽  
Rrna Gene ◽  
16S Rrna Gene Sequences ◽  
Pcr Assay ◽  
Nucleotide Sequence Data ◽  
The 16S Rrna Gene

Rapid molecular PCR-based detection method for Bacillus species used in the production of Beyonic® liquid organic fertilizer was carried out based on nucleotide sequence data from the 16S rRNA gene. The method involved sequencing the 16S rRNA gene of several Bacillus species and identifying around 16-22 specific nucleotide bases from 5' and 3' ends in the Bacillus 16S rRNA gene sequences. One specific primer pair for Bacillus detection was determined as follow: 5' - CAT AAG ACT GGG ATA ACT CCG GG - 3' (forward) from positions of 85-107 bp, and 5’ - CCA GGC GGA GTG CTT AAT GC - 3’ (reverse) from positions of 836-854 bp. PCR assay and gel electrophoresis analysis showed that the primer pair was specific to the genus Bacillus.

scholarly journals Zinc can counteract selection for ciprofloxacin resistance

2020 ◽  
Vol 367 (3) ◽  
Author(s):  
Michiel Vos ◽  
Louise Sibleyras ◽  
Lai Ka Lo ◽  
Elze Hesse ◽  
William Gaze ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Experimental Studies ◽  
Inhibitory Effect ◽  
Coli Strain ◽  
Resistance Evolution ◽  
Synergistic Interactions ◽  
Selection Dynamics ◽  
Fluoroquinolone Antibiotics ◽  
Ciprofloxacin Resistance ◽  
Selection For

ABSTRACT Antimicrobial resistance (AMR) has emerged as one of the most pressing threats to public health. AMR evolution occurs in the clinic but also in the environment, where antibiotics and heavy metals can select and co-select for AMR. While the selective potential of both antibiotics and metals is increasingly well-characterized, experimental studies exploring their combined effects on AMR evolution are rare. It has previously been demonstrated that fluoroquinolone antibiotics such as ciprofloxacin can chelate metal ions. To investigate how ciprofloxacin resistance is affected by the presence of metals, we quantified selection dynamics between a ciprofloxacin-susceptible and a ciprofloxacin-resistant Escherichia coli strain across a gradient of ciprofloxacin concentrations in presence and absence of zinc. The presence of zinc reduced growth of both strains, while ciprofloxacin inhibited exclusively the susceptible one. When present in combination zinc retained its inhibitory effect, while ciprofloxacin inhibition of the susceptible strain was reduced. Consequently, the minimal selective concentration for ciprofloxacin resistance increased up to five-fold in the presence of zinc. Environmental pollution usually comprises complex mixtures of antimicrobial agents. In addition to the usual focus on additive or synergistic interactions in complex selective mixtures, our findings highlight the importance of antagonistic selective interactions when considering resistance evolution.

Synthesis of novel 2-aminobenzothiazole derivatives as potential antimicrobial agents with dual DNA gyrase/topoisomerase IV inhibition

2020 ◽  
Vol 94 ◽  
pp. 103437 ◽  
Author(s):  
Magda M.F. Ismail ◽  
Hanan Gaber Abdulwahab ◽  
Eman Samir Nossier ◽  
Nagwan Galal El Menofy ◽  
Basma Abdelhameed Abdelkhalek
Keyword(s):  
Antimicrobial Agents ◽  
Dna Gyrase ◽  
Topoisomerase Iv

Prevalence and Antimicrobial Susceptibility of Salmonella Isolated from a Variety of Raw Meat Sausages in Gaborone (Botswana) Retail Stores

2012 ◽  
Vol 75 (4) ◽  
pp. 637-642 ◽  
Author(s):  
RONALD GAELEKOLWE SAMAXA ◽  
MAITSHWARELO IGNATIUS MATSHEKA ◽  
SUNUNGUKO WATA MPOLOKA ◽  
BERHANU ABEGAZ GASHE
Keyword(s):  
Antimicrobial Susceptibility ◽  
Salmonella Enterica ◽  
Antimicrobial Agents ◽  
Culture Method ◽  
Multidrug Resistant ◽  
Pcr Assay ◽  
Raw Meat ◽  
Conventional Culture ◽  
The Difference ◽  
Conventional Culture Method

The objective of the study was to provide baseline data on the prevalence and antimicrobial susceptibility of Salmonella in different types of raw meat sausages directly accessible to the consumers in Gaborone, Botswana. A total of 300 raw sausages comprising 79 beef, 78 pork, 72 chicken, and 71 mutton samples were concurrently analyzed for the presence of Salmonella using a conventional culture method and a validated PCR method. The PCR assay results were in full concordance with those of the conventional culture method for the detection of Salmonella. Sixty-five (21.7%) of 300 samples were positive for Salmonella by both the conventional culture method and PCR assay. Even though more chicken samples contained Salmonella than did any other sausage type, the difference in the presence of Salmonella among the four sausages types was not significant. Eleven serotypes were identified, and Salmonella enterica subsp. salamae II was most prevalent in all the sausage types. Beef sausages generally had higher mesophilic bacterial counts than did the other three sausage types. However, higher microbial counts were not reflective of the presence of salmonellae. Susceptibility of the Salmonella enterica serotypes to 20 antimicrobial agents was determined, and Salmonella Muenchen was resistant to the widest array of agents and was mostly isolated from chicken sausages. Regardless of the meat of origin, all 65 Salmonella isolates were resistant to at least four antimicrobial agents: amikacin, gentamicin, cefuroxime, and tombramycin. This resistance profile group was the most common in all four sausage types, comprising 90% of all Salmonella isolates from beef, 71% from pork, 63% from mutton, and 35% from chicken. These results suggest that raw sausages pose a risk of transmitting multidrug-resistant Salmonella isolates to consumers.

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