Phenotypic Detection and Biofilm Formation among Pseudomonas aeruginosa Isolated from Different Sites of Infection

2019 ◽  
Vol 10 (02) ◽  
Author(s):  
Ilham A Bunyan ◽  
Oday M Hadi ◽  
Hussein A K Al-Mansoori
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Virulence Factors ◽  
Biofilm Formation ◽  
Trichloroacetic Acid ◽  
Egg Yolk ◽  
Staining Technique ◽  
Protease Production ◽  
Clinical Samples ◽  
Bacterial Biofilms ◽  
Vitek 2

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.

Association between biofilm formation gene bla exoU and metallo and extend spectrum beta-lactamase production of multidrug resistance Pseudomonas aeruginosa in clinical samples

2021 ◽  
Vol 24 ◽  
Author(s):  
Fattma Abodi Ali
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Public Hospitals ◽  
High Rate ◽  
Clinical Samples ◽  
Antimicrobial Drugs ◽  
Product Size ◽  
Vitek 2 ◽  
Extend Spectrum Beta Lactamase ◽  
Control Management

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.

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
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Phylogenetic Trees ◽  
Antibiotic Sensitivity ◽  
Multidrug Resistant ◽  
Clinical Samples ◽  
Sensitivity Testing ◽  
Automated Method ◽  
Resistance Patterns ◽  
Vitek 2

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.

Investigate of the Ability of Cronobacter sakazakii Isolated from Clinical Samples of Children Under Two Years to Induce Swimming, Swarming and Biofilm

2018 ◽  
pp. 123
Author(s):  
Luma Abdal Hady Zwein ◽  
Tharieyt Abdulrahman Motlag ◽  
Mohamed Mousa
Keyword(s):  
Cerebrospinal Fluid ◽  
Congo Red ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Clinical Samples ◽  
Cronobacter Sakazakii ◽  
Yellow Color ◽  
Golden Yellow ◽  
Vitek 2 ◽  
Biochemical Examination

      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.  

scholarly journals Effect of tannic acid on urease and protease produced from Proteus mirabilis

2019 ◽  
Vol 13 (1) ◽  
pp. 82-86
Author(s):  
Omar S. Al-Zaidi ◽  
Luma abd alhady Zwein
Keyword(s):  
Tannic Acid ◽  
Biofilm Formation ◽  
Inhibitory Effect ◽  
Protease Production ◽  
Contaminated Water ◽  
Negative Bacterium ◽  
Urine Ph ◽  
Back Ground ◽  
Vitek 2 ◽  
Different Sources

Back ground: P.mirabilis is a gram negative bacterium, motile with its peritrichous flagella .Widely distributed in environment, especially in contaminated water and soil, Many virulence factors like LPS, urease, protease, hemolysin and biofilm formation play an important roles in the pathogenicity of P. mirabilis. Urease is a Nickel containing enzyme causes elevation of urine pH after hydrolyzing urea to ammonia and CO2 forming stones that blocks the urinary track. Aims: The effect of tannic acid on the production of urease and protease.                                                                        Material and methods: Twenty one isolates of Proteus were collected from different sources, Clinical and animal sources all isolates were cultured on MacConkey and blood agar and identification of P. mirabilis by, Vitek -2 compact system. Determine the effect of tannic acid on the production of urease and protease.                                                                                              Results: Twenty one isolates of Proteus were identified depending on Vitek-2 compact system, after identification, it turns out that only 18  isolate were P. mirabilis. All isolates were 100% able to produce urease and 72.2%  isolate were able to produce protease. The addition of tannic acid showed an inhibitory effect on urease and protease production. Conclusion: The effect of tannic acid on urease and protease depending on concentration, type of strain, incubation period, number of isolates and truculence of isolate.  

scholarly journals Evaluation of Antibiotic Resistance Pattern, Alginate and Biofilm Production in Clinical Isolates of Pseudomonas aeruginosa

2021 ◽  
Author(s):  
Fateme DAVARZANI ◽  
Navid SAIDI ◽  
Saeed BESHARATI ◽  
Horieh SADERI ◽  
Iraj RASOOLI ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Bacterial Resistance ◽  
Antimicrobial Agents ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Resistance Pattern ◽  
Alginate Production ◽  
Opportunistic Bacteria

Background: Pseudomonas aeruginosa is one of the most common opportunistic bacteria causing nosocomial infections, which has significant resistance to antimicrobial agents. This bacterium is a biofilm and alginate producer. Biofilm increases the bacterial resistance to antibiotics and the immune system. Therefore, the present study was conducted to investigate the biofilm formation, alginate production and antimicrobial resistance patterns in the clinical isolates of P. aeruginosa. Methods: One hundred isolates of P. aeruginosa were collected during the study period (from Dec 2017 to Jul 2018) from different clinical samples of the patients admitted to Milad and Pars Hospitals at Tehran, Iran. Isolates were identified and confirmed by phenotypic and genotypic methods. Antimicrobial susceptibility was specified by the disk diffusion method. Biofilm formation and alginate production were measured by microtiter plate and carbazole assay, respectively. Results: Sixteen isolates were resistant to all the 12 studied antibiotics. Moreover, 31 isolates were MultidrugResistant (MDR). The highest resistance rate was related to ofloxacin (36 isolates) and the least resistance was related to piperacillin-tazobactam (21 isolates). All the isolates could produce the biofilm and alginate. The number of isolates producing strong, medium and weak biofilms was equal to 34, 52, and 14, respectively. Alginate production was more than 400 μg/ml in 39 isolates, 250-400 μg/ml in 51 isolates and less than 250 μg/ml in 10 isolates. Conclusion: High prevalence of MDR, biofilm formation, and alginate production were observed among the clinical isolates of P. aeruginosa. The results also showed a significant relationship between the amount of alginate production and the level of biofilm formation.

scholarly journals Biofilm Formation and Virulence Factors Among Pseudomonas aeruginosa Isolated From Burn Patients

2015 ◽  
Vol 8 (10) ◽  
Author(s):  
Zahra Ghanbarzadeh Corehtash ◽  
Ahmad Khorshidi ◽  
Farzaneh Firoozeh ◽  
Hosein Akbari ◽  
Azam Mahmoudi Aznaveh
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Virulence Factors ◽  
Biofilm Formation ◽  
Burn Patients

Assessment of pelA-carried Pseudomonas aeruginosa isolates in respect to biofilm formation

2019 ◽  
pp. 1180-1187
Author(s):  
Mahmood Abd AL- Razzaq Hassan AL-Sheikhly ◽  
Laith N. Musleh ◽  
Harith J. F. Al-Mathkhury
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Nosocomial Infections ◽  
Biofilm Formation ◽  
Clinical Samples ◽  
Antibacterial Resistance ◽  
Gentamicin Resistance ◽  
Pcr Method

Owing to high antibacterial resistance of Pseudomonas aeruginosa, it could be considered as the main reason behind the nosocomial infections. P. aeruginosa has a well-known biofilm forming ability. The expression of polysaccharide encoding locus (pelA gene) by P. aeruginosa is essential for this ability. The purpose of the current research was to determine the biofilm formation in P. aeruginosa isolated from clinical samples and to evaluate the role of the selected PelA gene in biofilm formation using PCR method in Iraqi patients. Results revealed that 24 (96%) isolates were found to have the ability to form biofilm that was remarkably related to gentamicin resistance. Moreover, the pelA gene was found in all biofilm-producers. In conclusion, the results of the current study revealed that the P. aeruginosa biofilm-producer isolates were resistant to the antibiotics in question. Likewise, because of wide spreading, it appears that the pelA gene is related to biofilm formation.

scholarly journals Plantain Herb Extracts significantly attenuate the quorum sensing-controlled virulence factors and inhibit biofilm formation in Pseudomonas aeruginosa PAO1

2019 ◽  
Vol 78 ◽  
pp. 01004
Author(s):  
Shan Li ◽  
Jiangning Yao ◽  
Haoming Li
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Quorum Sensing ◽  
Virulence Factors ◽  
Biofilm Formation ◽  
Opportunistic Pathogen ◽  
Significant Inhibition ◽  
Control Group ◽  
Produce Cell ◽  
Group B ◽  
Herb Extracts

Pseudomonas aeruginosa is a Gram-negative organism that can survive under harsh conditions, and it is also an opportunistic pathogen that can produce cell-associated extracellular virulence factors. Several of these virulence factors have been demonstrated to be regulated by quorum sensing (QS). Plantain Herb has been used as antibacterial agents for many centuries in China. In this study, we analyzed Plantain Herb Extracts (PHE) at the concentration of 16 μg/mL (Group A, MIC), 8 μg/mL (Group B, 1/2 MIC) and 4 μg/mL (Group C, 1/4 MIC) for inhibition of the virulence factors production and biofilm formation in P. aeruginosa PAO1. The virulence factors included pyocyanin, rhamnolipids, protease and alginate. PHE showed significant inhibition of virulence factors as compared to the control group without interfering its growth. Thus, PHE might be a potent QS inhibitor and anti-biofilm agent in the treatment of Pseudomonas aeruginosa infections.

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