Plasmid Bands Research Articles

DETECTION OF PROTEUS SPECIES IN DIABETIC WOUNDS AND THEIR ANTIBIOTIC RESISTANCE PROFILE ANALYSIS

This work was carried out to determine the incidence of Proteus species infection in patients had diabetic foot wounds admitted to Vascular Surgery Department at Assiut University Hospitals. Proteus isolates were the most isolated organism (37.73%) followed by Klebsiella spp. (22.64%), then Pseudomonas spp. (20.75%) and E. coli (18.87%). Proteus mirabils was the most isolated species represented (78%) followed by Proteus vulgaris (13%) then Proteus penneri (9%). The antimicrobial susceptibility patterns of the isolated Proteus spp. were determined using agar disk diffusion method. The highest sensitivity was to ertapenem 76 isolates (76%). The antibiotic sensitivity then decrease in descending manner to be amikacin (65%) > meropenem (54%) > imipenem (52%) > cefipime (49%), while the highest resistance rates were to amoxicillin–clavulanic acid 75 isolates (75%) > co-trimoxazole (73%) > cefoxitin (63%) > ciprofloxacin (49%). Plasmid DNA profile analysis of 10 MDR Proteus mirabilis that were common resistant to ceftriaxone was studied. Plasmid bands of six resistant Proteus mirabilis isolates were shown at 800bp while the others at 700 bp. Plasmid curing was done by EtBr and SDS. Results of plasmid curing using ethidium bromide sublethal concentration of EtBr 1.25% showed that 7 cured cells become sensitive to ceftriaxone (30 µg), while 3 non cured cell still resistant to ceftriaxone. Plasmid curing using SDS sublethal concentration of SDS 1.2% cured only one of ten Proteus mirabilis which become sensitive to ceftriaxone and lost its band at 800bp, the other Proteus mirabilis not cured by SDS.

Plasmids and genes contributing to high-level quinolone resistance in Escherichia coli

Introduction: The importance of plasmid-mediated quinolone resistance (PMQR) in Enterobacterales and its high incidence has been emphasised many times. However, a clinical strain carrying more than two PMQR genes is rare. This study sequenced plasmid transconjugants from a donor strain carrying four different PMQR genes to establish their genetic locations.Methods: An Escherichia coli clinical strain displayed remarkable quinolone resistance with a ciprofloxacin MIC of 1024 mg/L carrying four PMQR genes: qnrA1, qepA1, aac(6′)1b-cr and oqxAB. When outcrossed to Escherichia coli J53 AziR, different PMQR genes were transferred and the resulting strains 7C and 8C were chosen for further characterisation. Plasmids were extracted and sequenced by the Illumina and Oxford Nanopore Technologies platforms. S1 nuclease-PFGE was used to estimate the number and size of plasmids.Results: The parental strain had three plasmid bands, as determined by PFGE. Transconjugant 8C obtained three plasmids: pMG336 (162 647 bp, F18:A-:B1:C4) carrying oqxAB; pMG335 carrying qepA1 (73 874 bp, F2:A-:B-); and pMG334 (59 724 bp, IncN (ST5)) with qnrA1 and aac(6′)1b-cr. Interestingly, strain 7C obtained plasmid pMG333 (134 435 bp), which was not present in the parental strain but was an IncN-IncF cointegrate of plasmids pMG334 and pMG335 linked via insertion sequence IS26.Conclusion: This study described the complete nucleotide sequence of three plasmids carrying four PMQR genes in a single strain and the plasmid profile obtained after outcrosses. In addition, it described a cointegrate of two plasmids formed with flanking insertion sequences.

Plasmid DNA Isolation and Visualization: Isolation and Characterization of Plasmids from Clinical Samples.

Plasmids are important in carrying antibiotic resistance and other genes between bacterial cells, and a number of methods can be employed to characterize plasmids from clinical isolates. Single colonies typically obtained as part of hospital workflow can undergo S1 nuclease treatment to linearize plasmids followed by pulsed-field gel electrophoresis to enable determination of the number and sizes of plasmids present. Hybridization of S1/PFGE gels can be used to associate replicon types and passenger genes, such as those conferring antibiotic resistance, with a particular plasmid band. Individual plasmids, obtained by conjugation or transformation, can be compared by gel electrophoresis following restriction digestion of plasmid DNA prepared by alkaline lysis methods, including using specialized kits.

Detection of multidrug-resistant Enterobacteriaceae isolated from river waters flowing to the Guanabara Bay and from clinical samples of hospitals in Rio de Janeiro, Brazil

The use of antibiotics in humans, animal husbandry and veterinary activities induces selective pressure leading to the colonization and infection by resistant strains. We evaluated water samples collected from rivers of the Guanabara Bay, which have suffered minor and major environmental degradation, and clinical samples of hospital origin to detect evidence of the presence of resistance genes to aminoglycosides, beta-lactam antibiotics and fluoroquinolones in strains of Klebsiella pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae and Escherichia coli. For isolation of the water strains we employed culture media containing 32 μg/ml cephalotin and 8 μg/ml gentamicin. The strains from clinical materials were selected using culture media containing 8 μg/ml gentamicin. The strains were identified and subjected to antimicrobial susceptibility testing (AST), plasmid DNA extraction and polymerase chain reaction (PCR) to detect genes encoding enzymes modifying aminoglycosides (EMA), extended-spectrum beta-lactamases (ESBL) and plasmid mechanisms of quinolone resistance (PMQR). The AST of the isolates recovered from water samples showed multidrugresistance profiles similar to those found in isolates recovered from clinical materials. All isolates from water samples and 90% of the isolates from clinical samples showed at least one plasmid band. In the PCR assays, 7.4% of the isolates recovered from water samples and 20% of those from clinical materials showed amplification products for the three antimicrobial classes. We believe that the detection of microorganisms presenting genetic elements in environments such as water is necessary for the prevention and control of their dissemination with potential to infect humans and other animals in eventual contact with these environments.

Isolation and Characterization of Plasmid - Bearing Multiple Antibiotic Resistance Bacteria from Different Aquatic Sources in Akure, Nigeria

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 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.

Influence of Helium-Neon Laser on Some Virulence Factors of Staphylococcus Aureus and Escherichia Coli

This work aims to investigate the effect of Helium-Neon laser on locally isolated Staphylococcus aureus & Escherichia coli bacteria; their resistance pattern, virulence factors, and their plasmid content. Bacteria were collected from patient suffering from urinary tract infections, septicemia, wound, and burn infections, then isolates were identified according to their characteristics, features; morphological, biochemical tests, and API systems. Out of Eighty-seven samples, twenty-seven isolates of S. aureus and thirteen isolates of E. coli were isolated. The results showed that after laser treatment, the diameter of inhibition zone increased for almost of the used antibiotics beside some isolates became sensitive especially after 2 min of laser exposure time. The hemolysin production was disappeared in (%40) of E. coli and (%20) of S. aureus after ten min of irradiation, while it decreased to half in another S. aureus isolate. All S. aureus and E. coli isolates loosed their ability to produce β-lactamase enzymes; some of these isolates loosed it after 5 minutes and the others after 10 minutes of irradiation. The results showed that laser irradiation hasn't any effect at any exposure time on adhesion factors of both S. aureus and E. coli isolates. Plasmid profile of Irradiated E. coli illustrated disappearing of DNA plasmid bands as well as RNA after ten min of irradiation.

A novel Trans-Pt(II) complex bearing 2-acetoxymethylbenzimidazole as a non-leaving ligand (trans-[Pt(AMBi)2Cl2]): Synthesis, antiproliferative activity, DNA interaction and molecular docking studies compared with its cis isomer (cis-[Pt(AMBi)2Cl2

Trans-[Pt(AMBi)2Cl2 (1) in which AMBi is 2-acetoxymethylbenzimidazole, was synthesized and characterized by spectroscopic methods. Novel complex 1 was evaluated for its potential antiproliferative effect against three human cancer cell lines, including HeLa (human cervix cancer), MCF-7 (human breast cancer) and A549 (human non-small-cell lung cancer) in comparison with those of cis-[Pt(AMBi)2Cl2 (2) and cisplatin by means of the MTT assay. Interaction of complexes 1, 2 and cisplatin with pBR322 plasmid DNA and their restriction endonuclease reactions by BamHI and HindIII enzymes were studied by agarose gel electrophoresis. Cytotoxicity tests showed that complex 1 exhibited similar in vitro cytotoxicity profile as compared with cisplatin whereas better than corresponding cis isomer against MCF-7 and A549 human cancer cell lines. The electrophoretic mobility shift assay showed that although no comigration of the Form I and II bands of plasmid DNA was observed for complex 1 at the concentrations tested, the binding of complex 1 was able to prevent partially BamHI digestion for concentrations ranging from 40 to 160 μM while having no inhibitory effect on HindIII activity in the digestion studies with restriction enzymes. Molecular docking studies on DNA were carried out to explore the binding mode and the best orientation of the complexes 1 and 2 to DNA.

Targeting of Virulence Factors and Plasmid Profiling of Klebsiella pneumoniae Causing Urinary Tract Infection in Sylhet City of Bangladesh

ABSTRACT Studies were conducted to characterize Klebsiella pneumoniae isolates from urinary tract infection (UTI) patients in Sylhet city of Bangladesh. At the same time, all isolates were screened for some common virulence genes and four significant isolates were searched for plasmid number and sizes by mini alkaline-lysis method. Among five tested isolates from female UTI patients, gyrase subunit B2 (gyrb2) amplified in all isolates, lipase and nuclease detected in three isolates and serine protease amplifies in two isolates and gave the expected band of 1130 bp, 517 bp, 1055 bp and 211 bp respectively. Two of four isolates showed 9.82 kb plasmid band on agarose gel. Isolates bearing 9.82 kb plasmid were found to be resistant to multiple commercial antibiotics. At the same time all isolates were screened for in-vitro plate assay for proteolytic, lypolytic and hemolytic activity. Isolates with positive plasmid and more than one virulent gene with gyrB2 showed positive result in in-vitro culture plate with clear zone of proteolysis, hemolysis or lipolysis. This study will be helpful for further study in finding correlation or pattern of virulence properties for K. pneumoniae associated UTI in Bangladesh.

Multi-antibiotics Resistant Relatedness of bla-gene Encoded Enteric Bacteria harbouring High Molecular R-plasmids.

Increase prevalence of multi-resistant enteric bacteria isolates encoded with high mobile R-plasmid causing enteric infections was examined among the community residents in Abeokuta, Nigeria. Random cluster sampling of 251 fecal samples of community residents were cultured for enteric bacteria and biotyped. Disc diffusion and Micro-broth dilution assay were used to determine antibiotic susceptibility while R-plasmid was profiled with photo-gel documentation. Antibiotic resistance relatedness was detected using DendroUPGMA construction utility software. Of all isolates obtained, 31.3% were Escherichia coli Klebsiella oxytoca (19.5%), Pseudomonas aeruginosa (15.3%) and Shigella specie (2.0%). Significant high rate of 62.6% showed resistant to Cefuroxime, 61.6% to Ampicillin and Augmentin (54.2%) while 44.7%, 38.9% and 33.9% resist Cotrimoxazole, Ciprofloxacin and Tetracycline respectively at MIC ≥16 μg/ml (p = 0.004). Only 54.1% harboured high molecular weight R-plasmid (>11.0kbp) and 2.7% having <5kbp R-plasmid weight. Two distinct clusters revealed significant multi-antibiotic resistant relatedness. Cluster A enteric isolates harboured similar R-plasmid of only one bands with high molecular weight more than 11kbp while Cluster B divided into subgroup a and subgroup b comprising different enteric species having similar high molecular weights with high antibiotic resistant expressing more than two plasmid bands showing computed cophenetic correlation of 0.94. Cluster analysis reveal a related high level multi-antibiotics resistant enteric bacteria strains among the community residents suggesting a continuous dissemination and imminent outbreak of resistant enteric pathotypes with resultant epidemic proportion.

Isolation and Identification of Burkholderia mallei and its Susceptibility to Anethum graveolens Extracts

In the present study, 13 (4.06%) isolates of Burkholderia mallei were obtained from hands of 320 workers in veterinary laboratories in Erbil Province- Iraq. All isolates identified depending on conventional assays, VITEK 2 compact system, and further confirmed by PCR using 23 rDNA gen. The results of PCR product showed that the expected size (526bp) was obtained for all local isolates and for B. mallei ATCC 15310. Also the effect of 10 antimicrobials against isolated bacteria were studied and the results showed that isolated B. mallei were absolutely resistant to norfloxacin, and 100% sensitive to imipenem and ceftazidime. The antibacterial activity of aqueous and alcoholic extracts of Anethum graveolens against B. mallei was studied. The results of MIC by using broth dilution method revealed that the alcohol and aqueous extracts exhibited inhibition of B. mallei growth and the MIC was 800, 1000 µg/ml, respectively. On molecular study, the SubMIC (750, 950 µg/ml) for both extracts against plasmid DNA profile of most resistant isolate of B. mallei was studied. The results showed three plasmid DNA band exist in local isolate of B. mallei and stander strain of B. mallei ATCC 15310 before treating with A. graveolens extracts. While it was decreased from three to two in the local isolate and the standard strain ATCC 15310 when treated with A. graveolens extracts. The inhibitory effect of the above mentioned extracts against total protein was studied by SDS-PAGE binding pattern. The results showed that there were differences in the protein banding pattern among the tested isolates and induction of some new protein bands when treated with plant extracts.

Detection Antibiotic Resistance of Enviromental Bacterial Strains

Antibiotics are randomly prescribed for veterinary and human medication. Antibiotics by little number are used by human , animals are digested uncompletely in their digestive system and ended up in communal sewage and hospitals, eventually discharge in environmental water sources directly with no processing.&#x0D; Water itself consider as major factor of dispersal of bacteria between different environmental components. Besides, bacteria had transferable genetic mobile elements to different sites of soil, water and humans.&#x0D; Environmental swabs were collected locally including 50 swabs of hospital environment , 15 samples of poultry feces and chicken guts , 20 sample of heavy water and 15 sample of fish tank to identify16 isolate of Staphylococcus (4 isolate of Staphylococus aureus and 12 isolate of coagulase –ve Staphylococcus) , 19 isolate of Enterococcus spp. , 7 isolates of Pseudomonas and 5 environment isolates for each Shigella spp. and Salmonella spp. . &#x0D; Teicoplanin and Vancomycin sensitivity test of isolates was done , showing that 2out of 16 isolates of Staphylococcus (12.5%) were Vancomycin-resistant , and 3out of 19 isolates of Enterococcus (15.7 %) were Vancomycin-resistant, while the rest of isolates were Vancomycin- sensitive. From other side , all isolates was Teicoplanin- sensitive except only 1 Enterococcus spp. Isolate which was intermediate . The range of the Vancomycin MIC were (6-64) µg/ml . Vancomycin resistant isolates , showed that some isolates have one plasmid band after Extraction of their DNA.

Characterisation of Staphylococcus aureus strains isolated from mastitis bovine milk in Argentina.

The study reported in this Research Communication was conducted to characteriseStaphylococcus aureusisolates recovered from mastitic bovine milk from dairy herds in Argentina. A total of 829 mastitic milk samples, both clinical and subclinical, were collected from 21 farms by veterinarians and submitted to the laboratory for testing from which 229S. aureusisolates were recovered, an isolation rate of 28·1%. These isolates were tested for susceptibility to the antibiotics penicillin, erythromycin and clindamycin. Of the 229 isolates, 53 (23·1%) were resistant to penicillin, 31 (13·5%) to erythromycin and 28 (12·2%) to clindamycin. All isolates were negative for themecA,mecC andpvlgenes by PCR. Southernblot hybridisation revealed that theermC gene was located on plasmid bands. Eighty isolates were randomly selected from the 229 for further characterisation. Restriction analysis of chromosomal DNA with Cf9I followed by PFGE of the 80 isolates revealed 23 distinct pulsotypes at 80% similarity. Seven major types (A, B, N, P, S, T, U and V) accounted for 68·7% of these isolates and 12 pulsotypes (A, B, F, G, J, K, M, N, P, S, T and U) occurred on more than one farm indicating genetic diversity within the farms. MLST of a representative isolate from dominant types identified the STs 97 705, 746, 2102 and 2187 with ST97 being the most predominant. Antibiotic susceptibility testing showed that 53·7% of the 80 randomly selected isolates were resistant to at least one of the three antibiotics tested. To our knowledge, this study represents the first large scale molecular studies onS. aureusisolates from dairy farms in Argentina.

Evaluation of biological potential of selected species of family Poaceae from Bahawalpur, Pakistan

BackgroundOxidative stress as well as bacterial and fungal infections are common source of diseases while plants are source of medication for curative or protective purposes. Hence, aim of study was to compare the pharmacological potential of seven grass species in two different solvents i.e. ethanol and acetone.MethodsPreliminary phytochemical tests were done and antioxidant activities were evaluated using ELISA and their IC50 values and AAI (%) were recorded. ANOVA was used for statistical analyses. DNA damage protection assay was done using p1391Z plasmid DNA and DNA bands were analyzed. Antimicrobial activity was done via disc diffusion method and MIC and Activity Index were determined. Cytotoxic activity was carried out using the brine shrimps’ assay and LC50 values were calculated using probit analysis program.ResultsPhytochemical studies confirmed the presence of secondary metabolites in most of the plant extracts. Maximum antioxidant potential was revealed in DiAEE, DiAAE (AAI- 54.54% and 43.24%) and DaAEE and DaAAE (AAI- 49.13% and 44.52%). However, PoAEE and PoAAE showed minimum antioxidant potential (AAI- 41.04% and 34.11%). SaSEE, DiAEE and ElIEE showed very little DNA damage protection activity. In antimicrobial assay, DaAEE significantly inhibited the growth of most of the microbial pathogens (nine microbes out of eleven tested microbes) among ethanol extracts while DaAAE and ImCAE showed maximum inhibition (eight microbes out of eleven tested microbes) among acetone plant extracts. However, PoAEE and PoAAE showed least antimicrobial activity. F. oxysporum and A. niger were revealed as the most resistant micro-organisms. ImCEA and ImCAE showed maximum cytotoxic potential (LC50 11.004 ppm and 7.932 ppm) as compared to the other plant extracts.ConclusionFodder grasses also contains a substantial phenols and flavonoids contents along with other secondary metabolites and, hence, possess a significant medicinal value. Ethanol extracts showed more therapeutic potential as compared to the acetone extracts. This study provides experimental evidence that the selected species contains such valuable natural compounds which can be used as medicinal drugs in future.

Detection of Sphingomonas paucimobilis and antibacterial activity of Prosopis farcta extracts on it

Six isolates of Sphingomonas paucimobilis were isolated from 120 hospital workers hands in Erbil city/Iraq by using VITEK2 Compact system, then further confirmed by PCR technique and by detecting specific gene TDP-glucose pyrophosphorylase (320bp) for S. paucimobilis ATCC 31461 and all local isolates. The effect of 17 antibiotics against isolated bacteria was studied and the results showed that all isolates were 100% sensitive to Piperacillin/Tazobactam, Imipenem, Meropenem, Ciprofloxacin, and Levofloxacin, while it was 83.33% resistant to Cefoxitin and Cefepime. The minimum inhibitory concentration (MIC) of Prosopis farcta pods extracts against S. paucimobilis isolate (S.p4) was 1000 μg/ml for methanol and ethanol extracts and 1200 μg/ml of watery extract. Moreover, the inhibitory effect of the above plant extracts (sub MIC) was studied against bacterial plasmid and the results showed three plasmid DNA bands when treated with 1000 μg/ml for watery extract and missing one band after treating with 800 μg/ml for both methanol and ethanol extracts. Also, the results of inhibitory effects of mentioned plant extracts on total protein by using SDS-PAGE showed the differences in protein banding pattern which 32 bands was formed when treated with 1000 μg/ml for watery extract and it was decreased to 16 bands when treated with 800 μg/ml for both methanol and ethanol extract.

Curing of plasmid contents of Proteus spp. isolated from urinary tract infections in AL- Diwaniyah city

This study included isolation and identification of Proteus spp . from urinary tract infections of Gynecology and padiatrics Hospital teaching in Diwaniyah cityand investigated the virulence factors connected with pathogenicity of this bacteria. It aims also at curing of plasmid contact by means of using some chemical materials. The bacteria Proteus spp. Isolates formed 16% which belong P. mirabilis and P. vulgaris from bacterial causes of urinary tract infection in children. The isolates of this bacteria has some physiological characteristics which represent virulence factors related to the ability of bacteria to make infection . It seems that 80% from these isolates were capable to Haemagglutination of RBCs . These isolates produce also Haemolysine enzyme. It was found that percentage of production was 60% . As for Blactamase enzyme production 40% of these bacteria isolates showed ability to produce his enzyme. Moreover, only 20% from Proteus isolates showed ability to produce bacteriocin production. All these isolates were not able to produce capsule production. When testing the resistance of Proteus isolates toward 8 different commonly used antibiotics in treatment of urinary tract infection, results showed that the isolates have different resistance depending on the type of antibiotic and the strain of the isolates. All these isolates were completely resistance Nitrofurantion in 100% while the least resistance toward Nalidixic acid and Gentamicin reached 20% for both. The results showed that there were DNA plasmid in 3 bacteria isolates but there was no relationship between the plasmid bands and resistance toward antibiotics. The P. mirabilis isolate which contained 4 plasmid bands was treated with SDS and Ethidiun bromide in an attempt to curing plasmid bands. The result showed that this isolate lost the ability to produce the majority of virulence factors that caused this disease and lost its resistance for som antibiotics especially Nalidix acid , Ampicillin and Choramphenicol. The ethidium bromide was the most effective in curing process since the resultant strain aest all plasmid bands.

Co-occurrence of colistin-resistance genes mcr-1 and mcr-3 among multidrug-resistant Escherichia coli isolated from cattle, Spain, September 2015.

Colistin resistance genes mcr-3 and mcr-1 have been detected in an Escherichia coli isolate from cattle faeces in a Spanish slaughterhouse in 2015. The sequences of both genes hybridised to same plasmid band of ca 250 kb, although colistin resistance was non-mobilisable. The isolate was producing extended-spectrum beta-lactamases and belonged to serotype O9:H10 and sequence type ST533. Here we report an mcr-3 gene detected in Europe following earlier reports from Asia and the United States.

The Effect of Coumarin Derivatives(compounds) on the Vibrio cholerae Isolates from Different Clinical Iraqi Sources

From a large number of bacterial samples collected from different hospital in Iraq in central health laboratory ,only ten isolates were identified primary as Vibrio. A number of morphology and biochemical test were carried out to complete this identification that showed all bacterial isolates were related to Vibrio cholerae .In this study all Vibrio isolates were investigated for Bio typing and the result showed that all (10) isolate were related to (Eltor biotypes) .Also, the susceptibility test towards eight antibiotics were carried out . Results shows that ciprofloxacin , Norfloxacin, Erythromycin, Ampicillin, ceftriaxone and Amikacin were the most effective antibiotics and their resistance percentage were 20%,20%,20, 20,30% and 30% respectively ,While Chloramphenicol and Co- trimoxazole were less effective and their resistance percentage were 90% both of them. Three (3,5,6) isolates V. cholerae were selected depending on results of antibiotics sensitivity tests as showed multiple –antibiotics resistance(100%). Then tested to study the effect of coumarin derivatives compounds (1, 2, 3 ) which showed inhibitory effect on V. cholera (3,5,6) isolates and the compound ( 3 ) showed the highest antibacterial activity of (12,15,14 mm) of inhibition zone diameter against V. cholera (3,5,6) isolates respectively. Also, these Iraqi isolates (3,5,6) used to test the effect of acridine orange (0.1%) as acuring agent , the results showed that all (3) isolates V. cholerae were sensitive to (ciprofloxacin, ceftrixone and Norfloxacin), While the rest were resistance to remained five antibiotics. The results of Agarose –gel electrophoresis of both normal V. cholerae (3,5,6) and cured isolates showed the presence of chromosomal and plasmid DNA bands in the normal case ,While only chromosomal DNA bands occur with V. cholerae (isolate 8) treated with an acridine orange at concentration of (10-2 to 10-4).

The Effect of Coumarin Derivatives(compounds) on the Vibrio cholerae Isolates from Different Clinical Iraqi Sources

From a large number of bacterial samples collected from different hospital in Iraq in central health laboratory ,only ten isolates were identified primary as Vibrio. A number of morphology and biochemical test were carried out to complete this identification that showed all bacterial isolates were related to Vibrio cholerae .In this study all Vibrio isolates were investigated for Bio typing and the result showed that all (10) isolate were related to (Eltor biotypes) .Also, the susceptibility test towards eight antibiotics were carried out .&#x0D; Results shows that ciprofloxacin , Norfloxacin, Erythromycin, Ampicillin, ceftriaxone and Amikacin were the most effective antibiotics and their resistance percentage were 20%,20%,20, 20,30% and 30% respectively ,While Chloramphenicol and Co- trimoxazole were less effective and their resistance percentage were 90% both of them. Three (3,5,6) isolates V. cholerae were selected depending on results of antibiotics sensitivity tests as showed multiple –antibiotics resistance(100%). &#x0D; Then tested to study the effect of coumarin derivatives compounds (1, 2, 3 ) which showed inhibitory effect on V. cholera (3,5,6) isolates and the compound ( 3 ) showed the highest antibacterial activity of (12,15,14 mm) of inhibition zone diameter against V. cholera (3,5,6) isolates respectively.&#x0D; Also, these Iraqi isolates (3,5,6) used to test the effect of acridine orange (0.1%) as acuring agent , the results showed that all (3) isolates V. cholerae were sensitive to (ciprofloxacin, ceftrixone and Norfloxacin), While the rest were resistance to remained five antibiotics.&#x0D; The results of Agarose –gel electrophoresis of both normal V. cholerae (3,5,6) and cured isolates showed the presence of chromosomal and plasmid DNA bands in the normal case ,While only chromosomal DNA bands occur with V. cholerae (isolate 8) treated with an acridine orange at concentration of (10-2 to 10-4).

Incidence of Methicillin-Resistant Staphylococcus aureus (MRSA) in Microbial Community of Cancer Patients and Evaluation of Their Resistant Pattern

Cancer patients are prone to MRSA infection due to a number of factors including prolonged hospitalization, intravascular catheterization, compromised host immunity, malignancy, chemotherapy, radiotherapy surgery, prior antibiotic therapy and prolonged operative time. Among 468 Staphylococcus isolates, 283 were methicillin-resistant Staphylococcus aureus isolates (MRSA), which indicated the occurrence and prevalence of MRSA isolates among cancer patients. The highest number of MRSA strains was isolated from male (124 isolates), female (90 isolates) followed by children (69 isolates), which represent 43.82, 31.80 and 24.38%, respectively, of all MRSA isolates. All S. aureus exhibited resistance to a higher number of antibiotics in common use. All isolates were oxacillin, cefoxitin (with MIC 256 to $${>}1024\,\upmu \,\hbox {g/mL}$$ ), clindamycin, penicillin, ceftriaxone, cephalothin and cefazolin (with MIC ranging between 128 and $${>}1024\,\upmu \,\hbox {g/mL}$$ ) resistant. 87.63% of MRSA isolates showed various multidrug-resistant profiles (MDRPs) toward more than one drug, and according to their MDRPs toward a panel of 17 antibiotics, MRSA strains were divided into seven patterns of resistance profiles. The existence of mecA gene corresponds to 100% with the methicillin phenotypic resistance in all MRSA isolates. The plasmid DNA profile of 85 multidrug-resistant S. aureus (MRSA) isolates showed that 42 isolates (49.41%) had plasmid bands.

Plasmid curing pattern of nosocomial pathogens of intensive care units (ICU) in Zagazig University Hospital

Nosocomial infections and antibiotic resistance are serious and growing phenomenon in contemporary medicine and has emerged as one of the public health concerns. A total of seventy isolates of bacteria were collected from patients. The selected isolates include 33 (56.9%) from males and 25 (43.1%) from females. All bacteria were susceptible to imipenem followed by amikacin, ceftazidime, ciprofloxacin and tobramycin, respectively. The multi-drug resistant (MDR) bacterial isolates (33 isolates) divided into four groups named Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus were selected to be identified according to the keys of different identification protocols. Escherichia coli found to be the most frequent pathogen within MDR isolates followed by Staphylococcus aureus, Pseudomonas aeruginosa and Klebsiella pneumonia, respectively. The MICs and MBCs of (Imipenem, Amikacin, Ceftazidime and ciprofloxacin) against the most resistant isolates were determined. Generally, P. aeruginosa (Strain No. 55) was found to be the highest resistant bacteria. Plasmid profile of Pseudomonas aeruginosa was carried out, after incubation at 37oc for 24hr recorded no changes in plasmid and compared with incubation at 43oc (plasmid curing) which showed complete removal of plasmid bands and the tested bacteria became more sensitive to antibiotics. This study therefore determined the prevalence, anti-biotic susceptibility and plasmid patterns of P. aeruginosa strains from clinical specimens obtained from the intensive care units of Zagazig University Hospital in Egypt.Asian J. Med. Biol. Res. December 2016, 2(4): 703-711