scholarly journals Evaluation of blaGES-5 and bla veb-1 genes with multidrug-resistant extend, pandrug resistance patterns (MDR, XDR, PDR), and biofilm formation in Pseudomonas aeruginosa isolates

2021 ◽  
Vol 67 (3) ◽  
pp. 52-60
Author(s):  
Fattma A. Ali ◽  
Sevan Hassan Bakir ◽  
Sayran Hamed Haji ◽  
Bashdar M. Hussen

Pseudomonas aeruginosa is a ubiquitous microorganism that is difficult to treat due to the increasing prevalence of multidrug resistance patterns. A total of 227 samples were taken from different clinical samples during the study period from January 2018 to December 2018. The isolates were identified with antibiotic sensitivity testing with ESBL by the Vitek-2 automated method. MDR, XDR, and PDR were determined. 40 (17.6%) isolates were positive for P. aeruginosa, maximum of ESBL and MBL. Positive isolates were detected in the burn, coexisting ESBL + MBL enzymes in 21 (52.5%) of our isolates. Imipenem followed by Meropenem were found to be effective against ESBL and MBL producers. Resistance was reached between 72-100% to 5 antibiotics. The frequency of PDR, MDR, and XDR were 5%, 50%, and 45%, respectively. The frequency of co-production between MDR, XDR, and PDR with MBL, ESBL, and Biofilm was 35%, 12.5% and 5%, respectively. Among the ESBLs, the frequency of distribution of bla VEB-1gene and blaGES-5 gene was 50% and 40 %, respectively. Bacterial isolates simultaneously carrying blaVEB-1 gene with multiple ?-lactamases of different classes of biofilm, MDR, PDR, and XDR as same as a coexisting blaGES-5 gene. One isolate was detected as new isolates registered in global gene bank as locally P. aeruginosa isolates in Erbil city (LOCUS MN900953). The phylogenetic trees of the blaVEB gene isolates were demonstrated a genotype closely related to others, deposited in GenBank similar to the P. aeruginosa gene; gene sequencing revealed a 99% similarity with other isolates deposited in GenBank.

2022 ◽  
Vol 13 (1) ◽  
pp. 88-92
Author(s):  
M Swapna ◽  
G Sumathi ◽  
M Anitha

Background: Pseudomonas aeruginosa is one of the most prevalent nosocomial pathogens that cause a life-threatening infection. One of the important characteristics of P. aeruginosa is biofilm formation which leads to antibiotic resistance. Aims and Objectives: The aim of the study was to study the antibiotic resistance pattern of P. aeruginosa isolates and correlation with their biofilm-production. Materials and Methods: A total of 87 P. aeruginosa isolates from different clinical specimens were processed and confirmed by conventional microbiological methods as per standard methodology. Antibiotic sensitivity testing was done for all isolates. Biofilm producing isolates were identified by the microtiter plate method (MTPM). Results: Of 87 P. aeruginosa isolates, majority were from pus 33 (38%), followed by urine 26 (30%), sputum 19 (22%), body fluids 7 (8%), and blood 2 (2%). Biofilm producing isolates showed more resistance in comparison to non-biofilm producers. The observed difference between biofilm formation for multidrug resistant and susceptible isolates was found to be statistically significant. Conclusion: MTPM method was an effective test for detection of biofilm formation and was also able to verify biofilm production by P. aeruginosa. This indicated a higher propensity among the clinical isolates of P. aeruginosa to form biofilm and revealed a positive correlation between biofilm formation and antibiotic resistance. This indicates the need for testing of even susceptible isolates for virulence factors such as biofilm production.


Author(s):  
Ilham A Bunyan ◽  
Oday M Hadi ◽  
Hussein A K Al-Mansoori

In The present study, included the collection of (100) samples from different clinical sites. Clinical samples were collected from patients who were visit and admitted All-Hilla teaching hospital at the period from November (2017) to February (2018). Cultural, biochemical and VITEK2 system were used for identification, and depending on the VITEK2 system (VITEK-2 GN Kit), revealed that twenty one (21) Pseudomonas aeruginosa isolates were recovered, The percentage conformational identification of P. aeruginosa was performed using VITEK2 system of (21) P. aeruginosa was (99%). Nine(42.8%) samples were isolated from burns, 5(23.8%) samples from wound, 3(14.2%) from urine, 2(9.5%) from ear swab, and 1(4.7%) sample was isolated from both blood and sputum. The phenotypic detection of some virulence factors for all isolates were detected. Detection of capsule was done by using capsule staining technique was carried out for P. aeruginosa isolates; it was found that all P. aeruginosa isolates (100%) have a capsule surrounding the bacterial cell. Hemolysin production by P. aeruginosa was studied; it was found that 12(57.1%) isolates were able to produce extracellular hemolysin on blood agar. P. aeruginosa isolates were also investigated for their ability to produce siderophores. The results showed that 9(42.8%) isolates of P. aeruginosa were able to produce siderophores. Protease production by P. aeruginosa isolates was studied; it was found that all these isolates (100%) have this enzyme as appear as a zone around the colony when being grown on (M9) media after adding of (3ml) of (5%) Trichloroacetic acid and incubation for (24 hrs.). Ability of P. aeruginosa to produce lipase has been investigated; it found that all these isolates (100%) were able to produce lipase after incubation for (48 hrs.) on egg yolk agar. Also, bacterial biofilms cause chronic diseases that are difficult to control and in the present study, differentiation of bacteria as biofilm producers and non-biofilm producers was done by using (ELISA) TCP method, a total of (21) isolates were tested for their ability to produce biofilm. From these isolates, (19) isolates were form strong biofilm, (2) isolates were form moderate biofilm.


Author(s):  
Fattma Abodi Ali

Background: The presence of biofilm formation exoU gene is significant challenge to infection control management in hospitals and exposure by Pseudomonas aeruginosa may lead to further spread and development of antimicrobial resistance. Methods: Out of 227 samples 40 clinical isolates of P. aeruginosa were collected from patients attending public hospitals ( Rizgary, Teaching hospital, Laboratory center, Raparin, Nanakaly hospitals)in Erbil city/Iraq over a period during 2018 to march 2019 and fully characterized by standard bacteriological procedures and antimicrobial susceptibility test and ESBL has been carried out by Vitek 2 compact system and. by Vitek 2 compact system. The identification has been verified by all isolates as P. aeruginosa by using 16S rDNA with product size (956pb). Results: A high rate of resistance was seen against Penicillin and lincomycin and Piperacillin and chloramphenicol and rifampicin (100 %), whereas Imipenem (5%) were found to be the most effective antimicrobial drugs. Of all P. aeruginosa isolates, 30 (75% %) were identified as MDR, approximately 9(22.5%) of isolates were resistant to 9 drugs in burn samples. Quantitative biofilm determination using the Congo red method revealed that 28 isolates (70%) produced biofilm, biofilm production was significantly higher among MDR P. aeruginosa isolates while coproduction of Extended Spectrum β-lactamase (ESBL) together with Metallo β-lactamase (MBL) ESBLs MBLs recorded in (52.5%) of the isolates. Altogether 40 isolates were processed for analysis by PCR assays and showed that 26(70%) of P. aeruginosa isolates harboured the exoU encoding gene with product size (204) pb was more commonly seen in isolates obtained from burn isolates. In addition, exo U gene was significantly associated with the higher MDR (80%), 8 isolates (76.9%)had exoU gene with ESBL and( 65%) had MBL and the same for MDR (80.8%) in samples for burning. Conclusion: Our results showed surveillance of P. aeruginosa resistance against antimicrobial and ESBL and MBL is fundamental to monitor trends in susceptibility patterns and appropriately guide clinicians in choosing empirical or directed therapy.


Author(s):  
Jitendra Kumar Chaudhary

The Pseudomonas aeruginosa is predominant agent causing nosocomial infections. In recent time, it develops resistance continuously to the antibiotics becomes Multidrug-resistant P. aeruginosa. So, in cystic fibrosis patents it difficult to eradicate P. aeruginosa infections with antimicrobial treatment. Therefore, focus on alternative mechanisms for treating P. aeruginosa infections. On the basis of growth, morphological and biochemical characteristics, P. aeruginosa strains were isolated from the clinical samples in this work. After that the antibiotic sensitivity was performed and the Multidrug resistant Pseudomonas aeruginosa was identified according to CLSI standard guideline chart by measuring the zone of inhibition for P. aeruginosa. The isolated strains showed resistance against three or more antibiotics, considered as MDR Pseudomonas aeruginosa. By antibiogram pattern 51 showed Multi drug resistant strains out of 102 isolated strains. As P. aeruginosa abide to develop resistance to the antibiotics, the quorum sensing increased transcriptional regulator QscR might performs another target. Thus the prevalence of MDR strains of Pseudomonas aeruginosa was investigated on current study. The antibiogram pattern revealed 51 MDR strains of P. aeruginosa. In which the majority of strains exhibited resistance towards Piperacillin (98%), Ciprofloxacin (90%), Ofloxacin (90%), Levofloxacin (80%) and Tobramycin (60%).


2020 ◽  
Vol 7 (3) ◽  
Author(s):  
Ibrahim A Naqid ◽  
Amer A Balatay ◽  
Nawfal Rasheed Hussein ◽  
Kurdistan Abdullah Saeed ◽  
Hiba Abdulaziz Ahmed ◽  
...  

Background: Escherichia coli (E. coli) is one of the most common causative agents of bacterial infections. The emergence of multidrug-resistant E. coli is a major public health threat worldwide. Objectives: This study aimed to determine the antibiotic susceptibility profile of clinical isolates of E. coli from different samples. Methods: A total number of 454 clinical samples, including urine, wound, cervical swab, blood, semen, ascetic, and cerebral spinal fluid samples were collected from patients between January 2017 and February 2020. Then, E. coli was confirmed and susceptibility to different antibiotics was determined using the Vitek-2 compact system. Results: Escherichia coli isolates were more frequent in females (70.7%) than in males (29.3%). In the case of urine samples, E. coli was found to be highly susceptible to ertapenem (97.6%) and imipenem (96.4%) but resistant to ampicillin (87.8%). For wound and cervical swabs, E. coli was 100% resistant to ampicillin and cefepime but 100% sensitive to ertapenem and imipenem. It was found that E. coli isolates from blood samples were 100% resistant to ampicillin, ceftriaxone, and cefoxitin, and around 75% of them were sensitive to ertapenem, ciprofloxacin, and levofloxacin. Finally, E. coli isolated from other clinical samples were highly sensitive to ertapenem, imipenem, levofloxacin, nitrofurantoin, and cefazolin. Conclusions: Escherichia coli isolated from various clinical specimens showed differences in antibiotic sensitivity patterns, with high resistance to commonly used antibiotics. The most effective antibiotics against E. coli isolates were ertapenem, imipenem, and nitrofurantoin. However, the clinical isolates of E. coli displayed high resistance rates to ampicillin, ceftriaxone, and cefepime. Therefore, it is proposed to perform antibiotic sensitivity testing by physicians to select the most effective antibiotics.


Author(s):  
Florence Chijindu Ugwuanyi ◽  
Abraham Ajayi ◽  
David Ajiboye Ojo ◽  
Adeyemi Isaac Adeleye ◽  
Stella Ifeanyi Smith

Abstract Background Pseudomonas aeruginosa an opportunistic pathogen, is widely associated with nosocomial infections and exhibits resistance to multiple classes of antibiotics. The aim of this study was to determine the antibiotic resistance profile, biofilm formation and efflux pump activity of Pseudomonas strains isolated from clinical samples in Abeokuta Ogun state Nigeria. Methods Fifty suspected Pseudomonas isolates were characterized by standard biochemical tests and PCR using Pseudomonas species -specific primers. Antibiotic susceptibility testing was done by the disc diffusion method. Efflux pump activity screening was done by the ethidium bromide method and biofilm formation assay by the tissue plate method. Genes encoding biofilm formation (pslA & plsD) and efflux pump activity (mexA, mexB and oprM) were assayed by PCR. Results Thirty-nine Pseudomonas spp. were identified of which 35 were Pseudomonas aeruginosa and 4 Pseudomonas spp. All 39 (100%) Pseudomonas isolates were resistant to ceftazidime, cefuroxime and amoxicillin-clavulanate. Thirty-six (92%), 10(25.6%), 20 (51.2%), 11(28%) and 9(23%) of the isolates were resistant to nitrofurantoin, imipenem, gentamicin, cefepime and aztreonam respectively. All the isolates had the ability to form biofilm and 11 (28%) of them were strong biofilm formers. They all (100%) harboured the pslA and pslD biofilm encoding genes. Varied relationships between biofilm formation and resistance to ciprofloxacin, ofloxacin, cefixime, gentamicin, imipenem, and aztreonam were observed. Only 23(59%) of the Pseudomonas isolates phenotypically exhibited efflux pump activity but mexA gene was detected in all 39 (100%) isolates while mexB and oprM genes were detected in 91%, 92%, and 88% of strong, moderate and weak biofilm formers respectively. Conclusion Multidrug resistance, biofilm and efflux pump capabilities in Pseudomonas aeruginosa have serious public health implications in the management of infections caused by this organism.


Author(s):  
Luma Abdal Hady Zwein ◽  
Tharieyt Abdulrahman Motlag ◽  
Mohamed Mousa

      The study included 200 samples were collected   from   children  under two   years included (50 samples from each of Cerebrospinal fluid, Blood, Stool and Urine) from, Central Children Hospital and Children's Protections Educational Hospital. Isolates bacterial were obtained cultural, microscopic and biochemical examination and diagnosed to the species by using vitek2 system. The results showed there were contamination in 6.5% of clinical samples. The diagnosed colonies which gave pink color on the MacConkey agar , golden yellow color on the Trypton Soy agar and green color on the Birillent Enterobacter sakazakii agar and gave  a probability of 99% in the vitek 2 and were identified as Cronobacter sakazakii. The identification revealed of thirteen isolates: 6(46.16%) isolated from Cerebrospinal fluid samples, 7(53.84%) isolated from blood samples and not isolated bacteria from stool and urine samples. The results of the investigation of some virulence factors showed that all bacteria isolates were able to swimming with a diameter ranging (1-9 mm) and swarming with a diameter ranging (1-40 mm) and their  ability to biofilm formation  by using three methods. The results show the ability  of  isolates to form biofilm by using  Congo red media  methods where it is 12 (92.30 %) out of 13 isolated bacteria belonging to C. sakazakii  able to form biofilm on the Congo red media  which is 3 (23.07%) were  strong production  biofilm ,   8 (61.53%)  were intermediate  production  biofilm and  1 (7.69% ) were weak  biofilm formation , while the 1 (7.69%)  unable to form biofilm.  Tubes method were all isolates were able to form biofilm, it were found that 3 (23.07%)  isolates strong, and 8 (61.53%) intermediate  and 2( 15.38%)  weak biofilm formation. Microtiter plate method  gave 5 (38.46 %) isolates strong, 6 (46.15%) intermediate and 1 (7.69%) weak biofilm formation.  


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mojisola C. Hosu ◽  
Sandeep D. Vasaikar ◽  
Grace E. Okuthe ◽  
Teke Apalata

AbstractThe proliferation of extended spectrum beta-lactamase (ESBL) producing Pseudomonas aeruginosa represent a major public health threat. In this study, we evaluated the antimicrobial resistance patterns of P. aeruginosa strains and characterized the ESBLs and Metallo- β-lactamases (MBL) produced. Strains of P. aeruginosa cultured from patients who attended Nelson Mandela Academic Hospital and other clinics in the four district municipalities of the Eastern Cape between August 2017 and May 2019 were identified; antimicrobial susceptibility testing was carried out against thirteen clinically relevant antibiotics using the BioMérieux VITEK 2 and confirmed by Beckman autoSCAN-4 System. Real-time PCR was done using Roche Light Cycler 2.0 to detect the presence of ESBLs; blaSHV, blaTEM and blaCTX-M genes; and MBLs; blaIMP, blaVIM. Strains of P. aeruginosa demonstrated resistance to wide-ranging clinically relevant antibiotics including piperacillin (64.2%), followed by aztreonam (57.8%), cefepime (51.5%), ceftazidime (51.0%), piperacillin/tazobactam (50.5%), and imipenem (46.6%). A total of 75 (36.8%) multidrug-resistant (MDR) strains were observed of the total pool of isolates. The blaTEM, blaSHV and blaCTX-M was detected in 79.3%, 69.5% and 31.7% isolates (n = 82), respectively. The blaIMP was detected in 1.25% while no blaVIM was detected in any of the strains tested. The study showed a high rate of MDR P. aeruginosa in our setting. The vast majority of these resistant strains carried blaTEM and blaSHV genes. Continuous monitoring of antimicrobial resistance and strict compliance towards infection prevention and control practices are the best defence against spread of MDR P. aeruginosa.


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