Distribution Pattern and Physical Mapping of CCC Plasmid-Mediated Multidrug-Resistance in some G-ve Bacteria Recovered from Hospitals and Haemodialysis Units Wastewater at Al-Madinah Al-Munawarah (original) (raw)
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Journal of Taibah University Medical Sciences, 2009
Objective Investigation of the presence and distribution pattern of plasmid-linked antibiotic resistance amongst Gram-negative bacterial strains recovered from wastewater of hospitals and haemodialysis units. Drawing up a physical map of the most frequently distributed plasmids. Methods Representatives G-ve bacterial strains; 59 isolates, were chosen from a previous study for the same authors, as of the most resistant ones (resist more than 3 up to 9 antibiotics to > 100 μg/ml.). Miniprep protocol and gel electrophoresis technique were adopted for the detection and isolation of ccc plasmid DNA from the differentially-isolated G-ve bacteria. Restriction analysis using 11 DNA restriction enzymes and "Plasp" computer program against λ phage DNA digested with Hind III, for physical mapping of the most distributed plasmid. Results Results revealed the presence of seven different plasmids, distributed as 1-3 different types of plasmids in 68% (40/59) of the strains; chosen as representatives of the most resistant ones (resist more than 3 up to 9 antibiotics to > 100 μg/ml.). The detected plasmid sizes were estimated against HindIII-digested λ phage and found ranged from 36,844 to 2,027 bp. Preliminary physical mapping was constructed for the plasmid (23130 bp) that was the most distributed amongst more than 90% of the plasmid-bearing strains and was the only found singly. Conclusion The isolated, characterized and the mapped plasmid (s), suggest the possibility of high rate of both vertical and horizontal distribution of these plasmids amongst the isolated Gve bacterial Genera and species.
Antimicrobial Agents and Chemotherapy, 1985
Forty-one isolates of multiply resistant gram-negative bacteria causing infection in a urological unit of a Dublin hospital were collected during a 6-month period. Twenty-one isolates transferred multiple resistance to an Escherichia coli K-12 recipient in liquid matings. Serratia marcescens, Proteus morganii, Proteus vulgaris, and F. coli isolates harbored similar 120-megadalton IncC plasmids, whereas Enterobacter cloacae strains transferred a 160-megadalton plasmid of a different Inc group. Southern hybridization experiments were performed with purified fragments cloned from one IncC plasmid as probes. They were hybridized to plasmid sequences in total cellular DNA extracts, showing that the IncC plasmids were very closely related. This suggests that the same plasmid has transferred to different bacterial species in the hospital environment. groups C
Plasmid proving antibiotic resistance in Al-Kharj Province
2016
The current work aims at screening pathogenic bacteria resistant to different antibiotics, clarifying the existence of resistivity due to free plasmids or plasmids integrated to chromosomes, and then isolating the plasmid involved in such resistivity and determining the precise antibiotic to which the plasmid is resistant. E. coli (8 isolates), Salmonella enteritidis (4 isolates), Proteus mirabilis (2 isolates), Pseudomonas aeruginosa (1 isolate), klebsiella pneumonia (1 isolate) were isolated from patients suffering diarrhea, fever, abdominal pain, and headache at King Khaled Hospital in Al-Kharj province. Thirteen antibiotics were investigated against the sixteen pathogenic bacterial isolates revealing the presence of resistant and sensitive isolates, subsequently plasmid isolation was performed on the unique nine selected isolates and the detected plasmids in five of the nine investigated bacterial pathogens were purified and transformed into the XL-blue 1 competent cells. The sensitivity assay results reveal the resistance of all of the investigated isolates to each of the antibiotics, mupirocin and cloxacillin. However, gentamycin, aztreonum, amikacin and meropenem were effective against all the investigated bacteria. While, the plasmid profile showed the presence of high molecular weight plasmids in five of the nine investigated bacterial pathogens, namely: Salmonella enteritidis (1);Pseudomonas aeruginosa (2) and Proteus marbilis (2).The transformed plasmid possesses resistivity genes for penicillin and cephalosporin groups only. On the other hand, the resistivity for all the rest of the investigated antibiotics might be integrated on the chromosome. So, the misuse of antibiotics facilitates the possession of pathogen's multidrug resistance through Rfactor (plasmid) production. This resistance could then be permanent through the integration of the plasmid to the bacterial chromosome preventing the possibility of the future use of such antibiotic.
Isolation of a Novel Antibiotic Resistance Plasmid DNA from Hospital
Emergence and dissemination of antibiotic resistance plasmids are major concern for hospital care system and increases the cost and decreases effectiveness of available antibiotics used in treatment of hospitalized patients. In this study two Pseudomonas aeruginosa, two Escherichia coli, and a Klebsiella pneumoniae were isolated from Intensive Care Unit (ICU) of a university hospital, in Kerman, Iran. K. pneumoniae exhibited resistance to all antibiotics routinely used in our hospital for treatment of patients except meropenem, while, the other isolates were sensitive to carbapenems and ciprofloxacin. Plasmid analysis of the selected isolates showed the presence of a single plasmid with molecular weight 65Kb in the P. aeruginosa isolate1. The plasmid was named as pKUM and belonged to incompatibility group -2 (IncP-2). Conjugation by filter mating revealed that resistance associated with gentamicin, kanamycin, cefotaxime, and ceftazidime phenotypes were transferred to E. coli ATCC25922 (Rif r ) recipient cells at the frequencies of 3.13 × 10 -5 and 5.3 ×10 -7 respectively. The results were further supported by curing and transformation experiments. MIC to cefepime was ≥30 μg/mL both in donor as well as the transconjugants while decreased to 0.5µg/ mL in cured derivative. The plasmid pKUM was quite stable (86%) in both donor cells and the recipient. From above results we concluded that resistances to third generation of cephalosporins, aminoglycosides and cefepime in P. aeruginosa isolate1 were indeed encoded by a conjugative plasmid. Acquisition of cefepime resistance through plasmid complicates the therapy of neutropenic patients in ICU and increases the cost and mortality of these patients.
Antibiogram and plasmid profile of E.coli isolates. International Journal of Bioresearch. 1:1-7.
The antibiotics sensitivity patterns and plasmid profiling was done to find out the possible correlation between plasmids and antibiotic sensitivity pattern of E. coli isolates. Disc diffusion method was followed for antibiogram and plasmid DNA from E. coli isolates was extracted through Mini alkaline lysis by SDS. Antibiogram study revealed that Ampicillin and Streptomycin resistant E. coli was more prevalent in broiler isolates (80%). Sulphamethoxazole Trimethoprim resistant E. coli was found mostly in layer isolates (83.33%) and in broiler isolates (60%). E. coli isolates of calves and cattle were sensitive to Chloramphenicol (71.43%, 88.89% respectively) and to Ciprofloxacin (100%) while water isolates were highly sensitive to Tetracyclin (66.67%). Agarose gel electrophoresis showed a total of 24 different bands of plasmids ranging from 0.5 to 40 kb in size. The plasmids were distributed at random in the isolated E. coli strains and no remarkable relationship between antibiotic resistance patterns and plasmids could be established.
Microbiology Research Journal International, 2019
Aims: This study was designed to investigate the plasmid bearing multiple antibiotic resistant bacteria from different aquatic sources. Place and Duration of Study: This research work was carried out in Akure South Local Government Area of Ondo state, Nigeria between January and June, 2018. Methodology: The pathogenic bacteria associated with water samples collected from different sources in Akure, Nigeria were isolated and characterized. A total of 521 water samples were collected from sources such as wells, taps, streams, rivers, boreholes and rain. All the samples were subjected to presumptive, confirmed and completed tests to evaluate their microbiological quality. The microbial types in the samples were determined using standard microbiological Original Research Article Onifade et al.; MRJI, 27(4): 1-12, 2019; Article no.MRJI.48233 2 techniques. All isolates obtained in this study were subjected to antibiotic sensitivity analysis and screened for Beta-lactamase production (ESBL). Plasmid profile analysis of the resistance isolates was carried out using standard method. Furthermore, post-curing of the plasmid mediated antibiotic resistance isolates was performed and data obtained were analyzed and presented using analysis of variance. Results: Bacterial isolates such as Acinetobacter baumanni, Citrobacter freundii, Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhi, Salmonella typhimurum, Salmonella paratyphi, Shigella dysenteriae, Serratia marcescens, Proteus vulgaris and Vibrio cholerae were identified from the water samples. The isolate E. coli had the highest percentage distribution of 24.10% in well water and 26.19% in stream water while Salmonella species had the highest occurrence of 53.85% in rain water. The Beta-lactamase producing (ESBL) isolates were resistant to multiple antibiotics except Ciprofloxacin, Gentamycin and Pefloxacin that conferred antibacterial effect. Plasmid-gene profile analysis of the isolates revealed that S. typhimurium, K. pneumoniae, P. aeruginosa and P. vulgaris possess single plasmid each while only E. coli contain two plasmid bands. The post plasmid-curing antibiotic sensitivity test of the isolates revealed that the initial antibiotic resistance of the bacterial isolates were plasmid mediated. Conclusion: Findings from this study suggest the purification of water from these sources before consumption is important as most microbes found in these samples are potential pathogens that are capable of causing infectious diseases with multiple antibiotic resistant features.
Antimicrobial resistance and Plasmid profile of Vi
The present study was aimed to detect the presence of multiple antibiotic resistance, antibiotic resistance genes and plasmid profile of Vibrio alginolyticus isolated from seawater and sediment of different beaches in Malaysia. Forty five isolates, including 24 seawater and 21 sediment isolates of V. alginolyticus were tested against 14 antibiotics for the antibiogram profile and the presence of the plasmids. Polymerase chain reaction (PCR) was conducted to elucidate the presence of 7 antibiotic resistance genes including Streptomycin resistance (strB), β-lactamase resistance (blaP1), Chloramphenicol Resistance (floR), Tetracycline Resistance (tetA), Erythromycin resistance (ermB), Quinolone resistance protein (qnrA) and Aminoglycosides resistance (aac(3)-IIa). Antibiotic resistance studies revealed that in seawater isolates, the highest percentage of antibiotic resistant was obtained against erythromycin E and penicillin P (100%), whereas the lowest antibiotic resistant percentage was obtained from both chloramphenicol C and nalidixic acid NA (16.66 %). The sediment isolates of V. alginolyticus showed 100% resistance against both penicillin P and ampicillin AM and the lowest percentage was of gentamycin CN (0 %). There were 17 different antibiotic patterns were observed from the V. alginolyticus in this study. The plasmid size was ranged from 2.3 Kb to 21.6 Kb, while there was no detection of plasmid in19 isolates. The highest resistance gene percentage of seawater isolates was found to be ermB (91.66%) which was followed by blapl with 70.83% of resistance gene. The lowest percentage of resistance gene was floR with 16.66% of resistance gene. The highest percentage of resistance gene in seawater isolates was found to be tetA with 61.9% of resistance gene and the lowest percentage was obtained from floR which had 14.28% resistance gene. The finding of this study was showen high percentage of resistance genes in seawater than sediment isolates. These results suggest that the V. alginolyticus isolated from seawater and sediments observed in this study were pathogenic, and involved a source of antibiotic resistance genes that could be transmitted to other population of bacteria through mobile genetic elements.
Conjugative plasmids in multi-resistant bacterial isolates from Indian soil. J Appl Microbiol
2008
Aims: Determination of heavy metal and antibiotic resistance and presence of conjugative plasmids in bacteria isolated from soil irrigated with wastewater. Methods and Results: Composite soil samples were collected from Ghaziabad, Uttar Pradesh, India. Forty different bacteria were selected from nutrient agar and characterized by morphological, cultural and biochemical tests. All the isolates were tested for their resistance to different heavy metals and antibiotics. The DNA derived from multiple metal and antibiotic-resistant bacterial isolates was PCR amplified and plasmid-specific sequences (IncP, IncN, IncW, IncQ and pMV158-type) were analysed by dot blot hybridization. All isolates gave PCR products with trfA2 and oriT primers of the IncP group. These PCR products also hybridized with the RP4-derived probes. However, the samples were negative for all the other investigated plasmids as proved by PCR and dot blots. Conclusions: The presence of conjugative ⁄ mobilizable IncP plasmids in the isolates indicates that these bacteria have gene-mobilizing capacity with implications for potential dissemination of introduced recombinant DNA. Significance and Impact of the Study: The detection of IncP plasmids in all the bacterial isolates is another proof for the prevalence of these plasmids. We propose that IncP plasmids are mainly responsible for the spread of multiresistant bacteria in these soils.