Cetrimide Agar Research Articles

Association of Multidrug Resistance With Biofilm Formation in Pseudomonas Aeruginosa Isolated from Clinical Samples in Kirkuk City

Pseudomonas aeruginosa is one of the most prevalent nosocomial pathogenic microorganisms that cause life-threatening infections. The formation of biofilm is one of the important features that lead resistance to antibiotics. The study aimed to clarify the relationship between biofilm formation, antibiotic resistance, and pigment production P. aeruginosa. A total of 33 isolates have been obtained from 220 different clinical samples, 42% of isolates obtained from burn swabs,15% wound swabs,12% from urine samples, 12% ear swabs ,6% sputum samples, and 3% from bronchial washes. The highest prevalence of these bacteria in our study was found in isolates obtained from burn swabs. Pyocyanin and pyoverdine are frequently seen on Cetrimide agar. P. aeruginosa can be completely isolated on Cetrimide agar. The production of biofilms detected by microtiter plate method and the result distributed as, 22% of strongly, 50% moderately, 27% weakly, and 4% were non biofilm producer. The Kirby-Bauer method (disc diffusion susceptibility method) was used to measure antibiotic resistance, and the results revealed a correlation as 86% multidrug resistance were strongly biofilm former, 75% multidrug resistance were moderately biofilm former and 11% multidrug resistance were weakly biofilm producers. Conclusion, P. aeruginosa from clinical isolates has highly proportional capability to form biofilm and recorded a positive correlation with multidrug resistance, while this relation was not significant with pigment production.

Spatio-temporal variation of bacteria of the Vibrio and Pseudomonas genera in some underground water points developed in the locality of Ombessa (Department of Mbam-et-Inoubou, Center-Cameroon)

A study aimed at evaluating the seasonal variation of Vibrio and Pseudomonas in groundwater in the city of Ombessa, Center region of Cameroon was conducted from January to July 2021. The microorganisms sought were Aerobic Mesophilic Bacteria BHAMs, bacteria of the Vibrio and Pseudomonas genera. These bacteria were isolated from 10 groundwater points by the surface spreading technique for BHAM and the membrane filter method for Vibrio and Pseudomonas. Plate count agar was used for BHAMs, TCBS medium for Vibrio and Cetrimide agar for Pseudomonas. Some abiotic parameters such as temperature, pH, were measured using the usual techniques. The data obtained was analyzed using the appropriate software. Correlation and comparison tests between the variables were carried out. It shows that the water studied has a pH of 5.94 UC to 7.9 UC, the content of dissolved O2 reached 5 mg/L, and there are positive and significant correlations between the density of isolated bacteria and the levels of physicochemical variables. Two species of the genus Pseudomonas (aeruginosa and fluorescens) and four of the genus Vibrio (cholerea, alginolyticus, vulnificus and parahaemolyticus) were isolated, with variable abundance rates ranges from 194 to 898 CFU/100mL for Pseudomonas and from 58 to 683 CFU/100mL for Vibrio. The presence of these germs in groundwater can be explain by the proximity of sources of pollution. This water would be unfit for human consumption without prior treatment according to the World Health Organization standard.

Antibiotic susceptibility and biofilm formation of Pseudomonas aeruginosa isolated from clinical and environmental hospital samples

Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen and a model bacterium for studying virulence and bacterial social traits. While, it can be isolated in low numbers from a wide variety of environments including soil and water, it can readily be found in almost any human/animal-impacted environment. It is a major cause of illness and death in humans with immunosuppressive and chronic conditions and infections in these patients are difficult to treat due to a number of antibiotic resistance mechanisms and the organism’s propensity to form multicellular biofilms. One hundred twenty clinical samples and forty hospital environmental samples (various sources) were collected from hospitals in Baghdad city during the period from October 2020 to January 2021, Cetrimide agar, the differential medium was used to isolation and diagnosis of Pseudomonas aeruginosa and 76 (63.33%) clinical isolates and 8 (20%) environmental samples were able to grow, and diagnosedas P. aeruginosa using the conventional biochemical tests, the Api 20 system and 16S rRNAgene. The antibiotic susceptibility test for fourteen antibiotics was performed by the standard disk diffusion method; the results showed that the bacteria were resistant to the most of the antibiotics used in this study and have the ability to form the biofilm.

Prevalence of blaIMP-1 among P. aeruginosa isolated from patients with burn infections

Carbapenems are often used as antibiotics for treating infections by multidrug-resistant Gram-negative bacteria. Pseudomonas aeruginosa is considered a major cause of health care issues associated with higher mortality and morbidity rates worldwide. This study aimed to detect the prevalence of carbapenemase gene blaIMP-1 that is produced by carbapenem-resistant P. aeruginosa isolated from burn infection patients. A total of 137 burn swabs were collected from burn patients in different hospitals in Babylon province from December 2021 to April 2022. All of the swabs were streaked on selective media cetrimide agar. Antibiotic susceptibility was performed for Imipenem and meropenem and polymerase chain reaction for species specific 16S rDNA gene and blaIMP-1 gene was done for all isolates. 50 isolates were confirmed by amplicon of species-specific 16S rDNA for P. aeruginosa. Results of antibiotic susceptibility for imipenem and meropenem revealed that 5/50 (10%) and 13/50(26%) of P. aeruginosa isolates were resistant to imipenem and meropenem, respectively. Co-susceptibility (co-resistance or co-sensitive) for both imipenem and meropenem results showed 5/50 (10%) display resistance for both imipenem and meropenem, 8/50(16%) display resistance for only meropenem while 37/50(74%) sensitive for both and none was resistant to imipenem and sensitive for meropenem. Phenotypic detection of Metallo-β-lactamase gave inverse result while the Mean±SD of inhibition zone (mm) for imipenem disc (10μg) =29.34±7.11 and for imipenem+EDTA (10 μg+750 μg) =21.8±4.98. Investigation of blaIMP-1 gene revealed that 23/50 (46%) have blaIMP-1 while 18/50 (36%) were sensitive for imipenem and meropenem but have blaIMP-1. The current study concludes the possibility of the presence of resistance gene among phenotypically sensitive isolates pushing great threat of antibiotic misuse and leaving the infection untreated.

Determination of Total Microbial Bioload and Pathogen Testing from Bakery Products

Bakery products are the most widely consumed food on the planet. Microbiological spoilage is one of the most common reasons for bakery products having a short shelf life. This study reports on isolation of microorganisms from bakery products. The study aimed to prospect the isolation of bacteria, total colony count and pathogen testing were carried out nutrient agar media and specific media. Isolation of bacteria was done using nutrient agar medium, total colony count was prosecuted by colony counting method and pathogen testing was performed using specific media like mannitol salt agar, EMB agar, cetrimide agar and bismuth sulphite agar. Biochemical characterization was executed for the obtained pathogen. It was preceded by test like indol test, MRVP test, simmon’s citrate test, catalase test, gelatinase test and TSI test. After completion of these tests it was concluded the obtained microorganism was Salmonella species.

AN ASSESSMENT OF THE MICROBIOLOGICAL QUALITY OF ANTI-VIRAL CHURNA SOLD IN INDIA BY AYURVEDIC MEDICAL PRACTITIONERS

Objectives: The objectives behind of this study are to analyze the unregistered herbal products dispensed by Ayurvedic practitioners, as per the WHO guidelines for determination of potentially hazardous contaminants. To aware the registered medical practitioners regarding contamination of herbal product, the patients are using unregistered herbal products from registered medical practitioners. Methods: The method employed in this study was a pour plate method. The specific media plates were incubated for 24 h at 37°C. On a colony counter, the plate was placed and observed for the presence of micro-organisms. The media used were nutrient agar medium and cetrimide nutrients, agar, saline agar, and MacConkey agar. Result: The present study found microbial contamination in herbal products that are widely available across the country. The formulations coded AV5, AV6, and AV7 were found to be contaminated by Pseudomonas aeruginosa and AV5 and AV6 were found to be contaminated by Escherichia coli, whereas the formulations coded AV1 and AV2 were found to be contaminated by Staphylococcus aureus in above permissible limit, posing a risk of infection if consumed. Conclusion: According to the findings of this study, it shows that few herbal formulations were showing the presence of micro-organism above the permissible of given by the WHO.

Draft genome analysis of a multidrug-resistant Pseudomonas aeruginosa CMPL223 from hospital wastewater in Dhaka, Bangladesh

Multidrug-resistant (MDR) clones of Pseudomonas aeruginosa can cause complicated infections in human. The emergence of ST664 of MDR P. aeruginosa has been reported in Nepal, Iran and China. Here, we present the draft genome analysis of a MDR P. aeruginosa CMPL223 isolated from hospital wastewater in Bangladesh to understand antimicrobial resistance trends and pathogenicity. Cetrimide agar was used for isolation of P. aeruginosa. Polymerase chain reaction (PCR) was carried out for detection of biofilm and integron related genes. Bacterial susceptibility to antibiotics was determined by disc diffusion method. Sequencing of whole genomic DNA was performed using Illumina iSeq 100 platform. Following quality checking of raw reads, assembly and annotation of sequences, a wide array of in silico tools were used for characterization of draft genome. The isolate was a strong biofilm former, carried integron 1 in chromosomal DNA, and was predicted to be pathogenic. It belongs to sequence type ST664 and O7 serogroup. The assembled genome contained 12 acquired antimicrobial resistant (AMR) genes, 2 prophage regions, 240 virulence genes, 71 drug targets, 142 insertion sequences, and 1 CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) array. The isolate was resistant to 21 out of 23 antibiotics, except colistin and imipenem. Comprehensive Antibiotic Resistance Database and ResFinder revealed that bacteria harboured blaOXA-50, blaOXA-796, blaPDC-374, fosA, tet(G), sul1, catB7, aph(3')-iib and ant(4')-IIb genes, conferring resistance to different classes of antibiotics. The results of in vitro characterization were consistent with the possible expression of detected antibiotic resistant genes through in silico analysis. Our data suggested the emergence of MDR P. aeruginosa ST664, which needs control measures for limiting its dissemination.

Molecular Characterization and Phylogenetic Analysis of Pseudomonas aeruginosa Obtained from Wound Infection

Purpose: Wounds infection is frequent hospital acquired infections which is caused by a wide variety of microorganism. Inappropriate treatment could be more hazardous to suffering patients and could lead to death. This manuscript is aimed to identify the existence of Pseudomonas aeruginosa from the clinical samples and determine the molecular evolution and population structure of Pseudomonas species using bioinformatics tools Method: The specimens were swiped from the bandage of infected patients, who were admitted for the therapeutic intervention in the hospitals, localized in Delhi-NCR region. Primarly, specimens were cultured in cetrimide broth for specific growth of Pseudomonas sp. Single colony isolation on cetrimide agar plate and characterization of bacterial colony by producing fluorescence under UV light was done to confirm Pseudomonas aeruginosa (P. aeruginosa). Furthermore, phylogenetic analysis was performed on the basis of PCR and sequencing of 16s rRNA sequence of P. aeruginosa. Results: Assay based on bacterial culture, biochemical and 16S rRNA gene analysis methods confirm the presence of P. aeruginosa in collected wound specimen. Basic local alignment search tool analysis indicates that newly sequenced 16S rRNA gene sequence was shown 99% similarity to P. aeruginosa species, analyzed using NCBI-BLAST tool. The phylogenetic analysis and nucleotide base composition studies performed using 45 sequences of 16S rRNA gene from 18 different species of Pseudomonas, including P. aeruginosa. The phylogenetic analysis was performed using Maximum Likelihood method for evolutionary relationships. Conclusion: The biochemical and molecular characterization revealed that the clinical samples were infected with P. aeruginosa. Further, phylogenetic analysis of 16S RNA sequences revealed evolutionary conservation among different Pseudomonas species.

Isolation and Identification of p-Nitrophenol Degrading Bacteria from Polluted Soil

Now-a-days due to an extensive industrialization, urbanization has leads concern towards organic pollution. Among the nitro-aromatic paranitrophenols are widely used as intermediates for the pesticides, dyes manufacturing processes. In the present study, a strain of Pseudomonas aeruginosa was isolated from the polluted soil especially in the vicinity of pesticide, chemical industry. This strain was able to utilize PNP as a sole source of carbon and energy by degrading it. The sub-culturing of these isolate on nutrient agar slants, growth on the Cetrimide agar formed well isolated colonies. The organism also gives bluish, greenish pigmentation on PIA (pseudomonas isolation agar). The organism is found to be gram-negative, exhibiting rods, motile. It is the catalase positive, MR positive, VP negative, Indole production is absent, it could utilize citrate as a sole source of carbon, it could rapidly liquefy gelatin and an oxidase negative. This studies have been showed that isolate NP1 have the ability to degrade the PNP. On the basis of the morphological, cultural and biochemical characteristics it was confirmed that the NP1 isolate have been identified as Pseudomonas aeruginosa.

Antibiotic susceptibility pattern of gram-negative bacteria isolated from apron and tables of meat vendors in Elele Market, Rivers State

Because of poor sanitary practices and a lack of knowledge about proper hygiene, meat vendors' aprons and tables could be potential sources of cross-infection of many public health bacteria. The purpose of this study was to determine the prevalence and antibiotic susceptibility of gram-negative bacteria isolates obtained from meat vendors' aprons and tables. 100 fresh swabs from meat vendors' aprons and tables were streaked on Eosin methylene blue, Cetrimide, and MacConkey agar and identified by the biochemical test. The antibiotic susceptibility pattern was evaluated on Mueller Hinton agar using the Kirby-Bauer disc diffusion method. Results: Out of the 100 samples processed, 66 Gram-negative bacteria were isolated. Klebsiella pneumoniae (74.24%) was the dominant gram-negative isolates, followed by E. coli (18.18%), Pseudomonas aeruginosa (4.55%), and Proteus sp (3.03%) respectively. The isolates from the tables recorded 100% resistance to Erythromycin and Cloxacillin with the highest sensitivity to Ofloxacin (66.7%) followed by Gentamicin (50%) while the isolates from aprons were 100% resistant to Augmentin and Cloxacillin followed by cefuroxime (93.3%) and Erythromycin (90%) with the highest sensitivity to Ofloxacin (100%) followed by Gentamicin (33.3%). This study confirmed the presence of multidrug-resistant gram-negative bacteria on the apron, and the levels of contamination detected on the aprons and tables used by meat vendors in Elele market presented an inadequate microbiological standard and quality. It is critical to implement proper orientation and workshops to educate meat vendors on standard practices to be followed when handling meat.

Evaluation the Efficacy of Bacteriophage Against Pseudomonas Aeruginosa Isolated from Wound and Burn Infections

One Hundred clinical samples were collected from different skin infection, including wounds and burns from patients in several hospitals in Baghdad governorate. The phenotypic diagnosis of isolated was performed by culturing on MacConkey agar medium and selective cetrimide agar. After a microscopic and biochemical tests diagnosis, the diagnosis was confirmed using Viteck2 system, 42 bacterial isolates were isolated belonging to P. aeruginosa, with ages ranging from (4 days-75 years). The results of sensitivity test with the Kirby–Bauer Disk diffusion technique found that 19 bacterial isolates only were highly resistant by 76.19% to Cefepime, while the resistance to Ciprofloxacin and Gentamicin was 45.2% for each of them. Moreover, it showed resistance to Aztreonem 33.33%, Ceftazidim and Piperacillin 28.5% each of them, while it also showed high sensitivity and low resistance to Imipenem 26.19%. The results revealed that 3 bacteriophages were isolated with different phenotypic characteristics and had an effective effect on isolates resistant to antibiotics and were studied under different conditions such as the change in temperature and incubation period, as well as the difference in glycerol concentrations.

The Prevalence Rate, Pattern of Antibiotic Resistance, and Frequency of Virulence Factors of Pseudomonas aeruginosa Strains Isolated from Fish in Iran

Pseudomonas aeruginosa (P. aeruginosa) is a pathogenic bacterium and one of the seafood's most common spoilage microorganisms. In this study, 470 fish samples collected randomly were evaluated for the presence of P. aeruginosa, antibiotic resistance, and frequency of virulence factors. Isolation of P. aeruginosa from fish samples was performed on cetrimide agar after an initial enrichment. Representative colonies were selected, and biochemical tests were conducted. An antibiotic resistance test was performed using the disk diffusion method. DNA was extracted, and antibiotic resistance genes, as well as virulence genes, were detected using PCR. Fresh fish showed the highest prevalence of P. aeruginosa (5%). No positive samples contaminated with P. aeruginosa were isolated from frozen fish samples. From smoked, salted, and dried fish, two samples (2.85–4%) were contaminated with P. aeruginosa. The antibiotic resistance against meropenem, imipenem, carbapenem, erythromycin, gentamicin, chloramphenicol, and enrofloxacin was 0%. The lowest antibiotic resistance pattern was observed in fresh fish, and the highest was observed in smoked, salted, and dried fish. Respectively, blaTEM, blaCTX-M, and blaSHV were the most abundant genes encoding antibiotic resistance. The most virulence genes were algD, algU, lasB, toxA, exoS, exoT, and apr. This study suggests that raw seafood could be a source of antibiotic-resistant P. aeruginosa and helps to spread resistance genes through the food chain. It seems that cross-contamination in the fishing, transportation, and supply of seafood can cause increased contamination with Pseudomonas aeruginosa in these products. Therefore, the hygienic principles can effectively reduce contamination by P. aeruginosa. Also, the prophylactic use of antibiotics in these products should be controlled.

Isolation and characterisation of carbapenem-resistant Pseudomonas aeruginosa from hospital environments in tertiary care hospitals in Dhaka, Bangladesh

Increasing evidence of carbapenem-resistant Pseudomonas aeruginosa (CRPA) infection in healthcare facilities poses an alarming threat to public health. There is little evidence on the occurrence of this organism in Bangladeshi hospitals. We collected 117 environmental swab samples from two tertiary care hospitals in Dhaka, Bangladesh and tested for Pseudomonas species by nonselective enrichment of swabs followed by plating on Cetrimide agar. We confirmed the isolates as P. aeruginosa by API 20NE test and polymerase chain reaction Polymerase Chain Reaction (PCR) for 16S rRNA gene. We analysed P. aeruginosa isolates for susceptibility against 15 clinically important antibiotics and tested the carbapenem-resistant isolates for metallo β-lactamase (MBL). All CRPA isolates were characterised for carbapenem-resistant genes, virulence genes and biofilm formation genes. Of 117 swab samples, 82 (70%) were tested positive for P. aeruginosa. All P. aeruginosa isolates were multidrug-resistant, and 39% (n=32) of isolates were CRPA. Around 56% (n=18) of CRPA were MBL-producing; 22% (n=7) of isolates were positive for carbapenemase gene blaNDM followed by 16% (n=5) for blaVIM and 13% (n=4) for blaIMP. Sequencing identified these genes as blaNDM-1, blaIMP-13, blaVIM-2 variants. Based on optical density values, 94% (n=30) of CRPA isolates were capable of producing biofilms. All CRPA isolates (n=32) were positive for at least 1 of 6 biofilm-associated genes and 4 of 12 virulence genes tested in the study. Hospital environments in Bangladesh are contaminated with highly virulent CRPA, which might be a potential source of hospital-acquired infections, accentuating the need for strengthening hospital infection control programs.

Molecular Technology for the Detection of Pyoviridine Gene in Pseudomonas aeruginosa Isolated from Burn Cases

Objectives: Using molecular technology for the purpose of confirmation of the diagnosis of Pseudomonas aeruginosa which depends on the detection of proteins like Pyoverdine gene of P. aeruginosa as diagnostic test. Materials and Methods: This work was done on 110 patients who had sustained burn injury from hospitals Baghdad, Al-Yarmouk and the Medical City Teaching Hospital during the period from October 2020 to the end of March 2021. The collected samples were cultured on different media (blood agar, MacConkey agar, nutrient agar, and Cetrimide agar) for isolation of P. aeruginosa bacteria as well as automated biochemical tests such as Api20E and VITEK 2 systems. The results showed that 76 (69%) samples had bacterial growth of P. aeruginosa. Antibiotic susceptibility testing was evaluated using the VITEK 2 compact. The Pseudomonas was detected using species-specific gene 16SrRNA gene using polymerase chain reaction (PCR) method and also detected by (PCR) using newly designed primers with a molecular size (389 bp) for pyoviridine gene. Results: This work showed that the predominant growth of burn wound infections was P. aeruginosa 76 samples (69%). Antibiotic susceptibility testing results showed the same sensitivity pattern of P. aeruginosa isolates to ceftriaxone and cephalothin (67.1%), gentamicin, piperacillin, ceftazidime, cefepime, and cefotaxime (65.7%). Resistance to imipenem, tobramycin, ticarcillin, and meropenem were (57.8%), (51.3%), (56.5%) and (55.2%) respectively, and had the highest sensitivity to amikacin (34.2%). Moreover, the highest resistance was to ciprofloxacin (69.7), and norfloxacin (71%). The results of using the 16SrRNA gene for the detection Pseudomonas give positive results (100%). Pseudomonas spp was detected by (PCR) for pyoviridine gene, revealed that 50% isolates give positive results. PCR product pyoviridine of the isolates (3) with the highest resistance to fluoroquinolones (ciprofloxacin and norfloxacin) was sent to nitrogen-based sequencing, and the sequencing results revealed the mutation presence. Conclusion: This study shows the current resistant pattern of P. aeruginosa against different classes of antibiotics and the involvement of several virulence genes in resistance mechanisms by using PCR which ultimately helps to select appropriate antibiotics useful for the treatment of many burned complicated by P. aeruginosa.

Effect Biofilm Formation in Pseudomonas aeruginosa Resistance To Antibiotic

Objective: Due Pseudomonas aeruginosa is the most common type of bacteria that causes hospital-acquired infections because it has multiple resistance mechanisms to antibiotics such as biofilm formation. It is important to understand to investigate biofilm production in multidrug resistant clinical isolates of P. aeruginosa from different sites of infection. Materials and Methods: This work was done on 114 patients were collected from different specimens From hospitals Baghdad. The collected samples were cultured on different media (blood agar, MacConkey agar, nutrient agar, and Cetrimide agar) for isolation of P. aeruginosa bacteria as well as isolates from all patient were tested for antimicrobial susceptibility using disk diffusion method, in vitro formation of biofilm in microtiter plates containing Muller –Hinton broth. Results: Forty specimens (35.1%) were observed to have bacterial growth (positive samples) for P. aeruginos, the results of P. aeruginosa isolates were tested for antibiotic susceptibility showed that among forty positive results, the most isolates were potentially resistant to different antibiotics presented that the isolates resistant to piperacillin (34, 85%), gentamicin (26, 65%), tobramycin (30, 82.5%), amikacin (27, 67.5%), cefepime (25, 62.5%), meropenem (31, 77.5%), carbenicillin (31, 77.5%), ceftriaxone (30, 75%), ciprofloxacin (27, 67.5%), imipenem (22, 55%), ceftazidime 26 (65%), and norfloxacin 24 (60%). The results showed the ability of P. aeruginosa isolates to produce biofilm were 40 (100%) has the ability to produce biofilm, these were the result divided in to strong 19 (47.5%), moderate 12 (30%), and weak 9 (22.5%). Conclusion: There were 40 isolates biofilm producers in divers levels of biofilm strength.

1246. Clinical isolates of Pseudomonas aeruginosa Harbor Preexisting Changes in TonB-dependent Receptors Associated with Decreased Susceptibility to Cefiderocol

BackgroundCefiderocol (FDC) is a novel siderophore cephalosporin that retains activity against MDR gram-negative bacteria. In P. aeruginosa (PA), FDC utilizes TonB-dependent receptors (TBDR) PirA, PiuA, or PiuD to enter the periplasmic space. We recently reported a clinical PA isolate that developed elevated MICs to FDC associated with mutations in genes encoding TBDRs in the absence of prior exposure to FDC. In this study, we investigated the frequency of TBDR mutations not associated with cefiderocol exposure among clinical stains of PA recovered from 1999 to 2018 in a large hospital system in Houston, TX. MethodsA total of 212 clinical isolates of PA were screened for mutations in TBDR pathways (pirA/pirRS and piuA/D) via whole genome sequencing. Strains with gene mutations predicted to significantly alter protein function (insertion, deletion, or frameshift) were selected for phenotypic characterization. PA PA01 and 4 clinical PA strains lacking changes in the TBDR genes served as controls. FDC susceptibility testing was performed on Mueller-Hinton agar by Kirby-Bauer disc diffusion (DD). Diameters were measured at 18 h and 48 h to assess for the emergence of colonies inside the zone of inhibition. Breakthrough colonies were isolated on cetrimide agar, and DD studies were performed to determine FDC susceptibility.Results6.1% of isolates (13/212) had preexisting mutations in the TBDR genes, including indels in piuA (n=2) and pirR (n=2), and a frameshift mutation resulting in premature stop codon in pirR (n=9). DD showed that isolates with predicted changes in TBDRs had a significantly smaller diameter of inhibition, as compared to controls (Fig 1). Of the PiuA or PirR mutants, 3 of 13 demonstrated breakthrough colonies (Fig 2); while none of the control specimens showed breakthrough colonies. Subcultures of isolated breakthrough colonies yielded more homogenous populations of PA with relatively lower DDs than the original strain (Fig 2). Figure 1 Cefiderocol disk diffusion diameters of P. aeruginosa isolates at (a) 18 hours growth and (b) 48 hours growth. P-values for student’s two-tailed t-test are given. Dotted lines represent cutoffs for intermediate and resistant per Clinical Laboratories and Standards Institute (CLSI) guidelines. ns, non-significant; TBDR, TonB dependent receptor.Figure 2 Disk Diameter resistance phenotypes suggestive of heteroresistance among P. aeruginosa strains containing TBDR mutations (row 1) and their subsequent breakthrough colony subculture resistance phenotypes (row 2) at 48 hour time points.ConclusionMutations in genes encoding TBDR are present in clinical isolates of PA that predate the approval of FDC and are associated with the emergence of reduced susceptibility to FDC.Disclosures Truc T. Tran, PharmD, Merck (Grant/Research Support) Cesar A. Arias, M.D., MSc, Ph.D., FIDSA, Entasis Therapeutics (Grant/Research Support)MeMed Diagnostics (Grant/Research Support)Merk (Grant/Research Support) William R. Miller, MD , Entasis Therapeutics (Scientific Research Study Investigator)Merck (Grant/Research Support) William R. Miller, MD , Entasis (Individual(s) Involved: Self): Scientific Research Study Investigator; Merck (Individual(s) Involved: Self): Grant/Research Support

Prevalence of Biofilm Genotype Pattern( algD −/pslD −/pelF –) with Multidrug-Resistant in Clinical Local Pseudomonas Aeruginosa Isolates

The study was designed to explore the distribution and association of the biofilm genotype pattern(algD −/pslD −/pelF –) with multidrug-resistant in clinical local Pseudomonas aeruginosa isolates. Atotal of one hundred isolates of Pseudomonas aeruginosa were included in this study, which has beencollected from different specimens, from July to September 2020. The isolates included were 34 fromburns, 19 from wounds, 23 from ear infections, 22 from urinary tract infections (UTI), and 2 from cysticfibrosis (CF). Identification of the isolates was carried out using microscopical, cultural characterizationon MacConkey agar, Cetrimide agar, then Pseudomonas agar. Biochemical tests were performed, andfurther identification was carried out by the VITEK_2_compact system. Genotypic identification hasbeen completed by16SrRNA. To assess the frequency of multidrug-resistant of Pseudomonas aeruginosa(MDR), the antibiotic susceptibility test was done. It was carried out by using different groups ofantibiotics (10 antibiotics) using the Kirby–Bauer disk diffusion method. The results showed that theresistance were Ceftazidime(62%),Gentamicin(26%,(Piperacillin-tazobactam(25%), Ticarcillin(24%),Meropenem(20%), Cefepime (18%),Amikacin(17%) Levofloxacin(16%), Colistin(15%) Imipenem(10%).Biofilm production was assessed using a microplate examination method. The results showed that 93%of isolates were positive for biofilm production, while (7%) were non-biofilm producers. There weredifferences in the rates of biofilm-production distributed into 21 (21%) were strong biofilm producer(OD was more than 2.156), 25 (25%) intermediate biofilm producer, and 47 (47%) were weak biofilmproducer (OD was less than 1.078), and the non -biofilm producer was 7(7%).Three virulence factors genes ( algD, pslD, and pelf ) were chosen, which responsible for the phenotypicpattern of biofilm formation and identified as genotypic algD −/pslD −/pelF – pattern. The differencesin genotypic pattern prevalence among the MDR-positive isolates of different origins were statisticallysignificant. Chi-square analysis showed a highly significant association between strong biofilm capacityand genotype pattern (p<0.0001), also the analysis showed a highly significant association betweenmoderate biofilm capacity and genotype pattern (p<0.002). Chi-square analysis showed a highlysignificant association between weak biofilm capacity and genotype pattern ( p<0.001).In the current study the percentage of resistance among P. aeruginosa local isolates for multiple antibiotics(MDR) was relatively low, maybe due to the combination strategies based on appropriate anti-pseudoantibioticagents that may be used to improve treatment from the related infections, according to theseresults, P. aeruginosa local isolates that produced biofilm were mostly (70%) indicated as non-MDR.

ISOLATION AND PRODUCTION OF POLY-?-HYDROXY BUTYRATES

Polyhydroxybutyrates (PHB) are bio-plastics produced by many microorganisms under carbon rich conditions. PHBs are carbon storage compounds also known as Poly hydroxyalkanoates.PHB’s are high in molecular weight, polyester is accumulated as a storage carbon in many species of bacteria and is a biodegradable thermoplastic. Several gram-negative bacteria, including pseudomonas aeruginosa, have been employed for the efficient production of PHB. These gram negative bacteria can release endotoxin(pyrogen), which was in the form of lipopolysaccharides, from the cell wall (Raetz, 1993). Since PHB was most efficiently produced by gram-negative bacteria as described, the endotoxin levels present in the purified PHB should be examined can accumulate PHB in the form of multiple granules of 0.2 to0.5μm size. Pseudomonas species contains high PHB accumulating ability. Pseudomonas species are Gram negative, rodshaped aerobic bacteria Bacteria were isolated from native environment by culturing on nutrient agar and cetrimide agar media. When stained with Sudan Black Dye black colour granules are observed As a positive result. Polybeta-hydroxybutyrate can be further used for the production of Bio plastics. Microbial biosynthesis of PHB starts with the condensation of two molecules of acetylcoenzyme to give acetoacetyl-CoA which is subsequently reduced to hydroxybutyryl-CoA. This latter compound is then used as a monomer to polymerize. The microbial production of copolymers of 3- hydroxybutyrateand 3- hydroxyvalerate, with properties varying according to copolymer composition, must be considered. The production cost of PHB’s are very high when compared. to non-degradable plastics hence search for potential strains that accumulates high PHB’s

APPLICATION OF PHENOTYPIC AND GENOTYPIC STRATEGIES TO IDENTIFY PSEUDOMONAS AERUGINOSA

A case study of patient presented with developed wound infection with Pseudomonas aeruginosa (leukocytosis). The strains of P. aeruginosa isolated from Sheffield General Northern Hospital (UK) and studied for phenotypic and genotypic identifications. This present study were conducted in Sheffield Hallam University (UK) and was aimed to evaluate the application of Phenotypic and Genotypic techniques to identify P. aeruginosa. Generally, the results of Real time PCR (RTPCR) revealed that the diagnosis viability was confirmed for P. aeruginosa. However, identification on the basis of phenotype by differences in types of growth on cetrimide and acetamide agar medium presents that both agars were conducive to the growth of P. aeruginosa. The strains of P. aeruginosa were also tested for antibiotic susceptibility to six different antibiotics, imipenem showed the greatest inhibition effect on the bacteria. It was observed that genetic techniques in accordance with phenotypic tests have facilitated to conduct a comprehensive characterization of P. aeruginosa strains obtained from a particular environment at a particular time.

Molecular Investigation of Quinolone Resistant Genes among Clinical Psedomonas aeruginosa Isolates

The current research was performed in Al-imam AL- Sadiq Hospital, Hilla Teaching Hospital, Babyloncity, Iraq to investigate the resistance gene of fluoroquinolones (FQs) in P. aeruginosa clinical isolatesof taken from different origins. Mutations arising at the bacterial gyrA, gyrB, parC, par E gene belong toDNA gyrase and topoisomerase IV are the fluoroquinolone tolerance essential mechanism in P. aeruginosa..and over expression of efflux pumps . (80) clinical isolates collected by Transport swabs from varioussource [(burn 45(56.25 %), wound 15 (18.75 %), ear 3 (3.75 %), blood 2 (2.5 %), urine 4(5 %),sputum11(13.75)] that initially identified by culturing on MacConkey agar, blood agar and cetrimide agar selectivemedia for Pseudomonas aeruginosa, P. aeruginosa chromogenic agar then diagnosed by performing somemorphological and biochemical tests. The second diagnosis to (10) isolates was done by VITEK 2 Compactsystem using VITEK 2GN,and by using PCR technique for genotyping detection by the 16s rRNA gene.A susceptibility test was applied to all (80) isolates using 9 antibiotics through resorting to Kirby-Pourmethod. The results of this test showed that isolates which represented the highest resistance to ciprofloxacin(71%),levofloxacin(75%) , aztreonam(87.5%), but isolates showed low resistance to impenem 18.75% and0%resistant to meropenem. (gyrA, gyrB) subunit of DNA gyrase is target of flouroquinolon antibiotic hasbeen screened by PCR reaction. The results showed that these genes present in P. aeruginosa at (100 percent,100 percent,), respectively..