Lactamase Enzymes Research Articles

Incidence of Klebsiella pneumoniae producing Metallo Beta-Lactamase (MBL) at RSUP Dr. Wahidin Sudirohusodo Makassar

Bacterial resistance to antibiotic is one of the factors triggering infection therapy failure. This study was conducted to determine the prevalence of carbapenem-resistance Klebsiella pneumoniae infection and the phenotype of carbapenem-resistant Metallo-Beta-Lactamase (MBL)-producing Klebsiella pneumoniae isolates at RSUP Dr. Wahidin Sudirohusodo Makassar. This study included Klebsiella pneumoniae identification on each infectious patient’s isolates. The sensitivity test of antibiotics, phenotype confirmatory test, and MBL phenotypic test were conducted using agar diffusion Kirby-Bauer, Vitek-2-Compact, and Double Disc Synergy Test (DDST) method, respectively. As the result, the antibiotic sensitivity test using the Vitek-2-Compact method on 50 clinical samples (pus, sputum, blood. tissue, urine, brain fluid, and feces) found that 10 isolates (20%) were resistant to carbapenem. The phenotypic test using the Double Disc Synergy Test (DDST) method found that carbapenem-resistant isolates caused by the production of Metallo Beta Lactamase (MBL) enzymes were 2 isolates or 20% of the total carbapenem-resistant isolates.

Cefotaxime Exposure Selects Mutations within the CA-Domain of envZ Which Promote Antibiotic Resistance but Repress Biofilm Formation in Salmonella.

ABSTRACTCephalosporins are important beta lactam antibiotics, but resistance can be mediated by various mechanisms including production of beta lactamase enzymes, changes in membrane permeability or active efflux. We used an evolution model to study how Salmonella adapts to subinhibitory concentrations of cefotaxime in planktonic and biofilm conditions and characterized the mechanisms underpinning this adaptation. We found that Salmonella rapidly adapts to subinhibitory concentrations of cefotaxime via selection of multiple mutations within the CA-domain region of EnvZ. We showed that changes in this domain affect the ATPase activity of the enzyme and in turn impact OmpC, OmpF porin expression and hence membrane permeability leading to increased tolerance to cefotaxime and low-level resistance to different classes of antibiotics. Adaptation to cefotaxime through EnvZ also resulted in a significant cost to biofilm formation due to downregulation of curli. We assessed the role of the mutations identified on the activity of EnvZ by genetic characterization, biochemistry and in silico analysis and confirmed that they are responsible for the observed phenotypes. We observed that sublethal cefotaxime exposure selected for heterogeneity in populations with only a subpopulation carrying mutations within EnvZ and being resistant to cefotaxime. Population structure and composition dynamically changed depending on the presence of the selection pressure, once selected, resistant subpopulations were maintained even in extended passage without drug.IMPORTANCE Understanding mechanisms of antibiotic resistance is crucial to guide how best to use antibiotics to minimize emergence of resistance. We used a laboratory evolution system to study how Salmonella responds to cefotaxime in both planktonic and biofilm conditions. In both contexts, we observed rapid selection of mutants within a single hot spot within envZ. The mutations selected altered EnvZ which in turn triggers changes in porin production at the outer membrane. Emergence of mutations within this region was repeatedly observed in parallel lineages in different conditions. We used a combination of genetics, biochemistry, phenotyping and structural analysis to understand the mechanisms. This data show that the changes we observe provide resistance to cefotaxime but come at a cost to biofilm formation and the fitness of mutants changes greatly depending on the presence or absence of a selective drug. Studying how resistance emerges can inform selective outcomes in the real world.

Prevalence of b-lactamase produced in Klebsiella pneumoniae and Enterobacter cloacae isolated from gingivitis in Al-Najaf Province, Iraq

Abstract. Motaweq ZY. 2022. Prevalence of ?-lactamase produced in Klebsiella pneumoniae and Enterobacter cloacae isolated from gingivitis in Al-Najaf Province, Iraq. Nusantara Bioscience 14: 78-83. This study provides phenotypic and genotypic ?-lactamase formation data on 26 isolates of Klebsiella pneumoniae and Enterobacter cloacae isolated from patients suffering from gingivitis checked at Al-Kafeel clinic and private clinic in Al-Najaf Province-Iraq during the period from September 2020 to February 2021. In this study, some of them were detected by traditional phenotypic methods, while others were detected by phenotypic and then genotypically by using monoplex-PCR technique. The results revealed that out of 14 ?-lactam resistance K. pneumoniae isolates, 8 isolates (57.1%) gave positive results with the direct capillary tubes method, while 12 ?-lactam resistance E. cloacae gave 6 (50%) positive results. This result indicated that the enzymatic resistance was prevalent among isolates. Furthermore, the results showed that most isolates were ESBL producers according to initial and confirmatory methods. Molecular amplification of the ?-lactamase enzyme blaSHV gene was detected in 8 (57.1%) and 5 (41.6%) for K. pneumoniae and E. cloacae, respectively. While all 100% of K. pneumoniae and E. cloacae isolates gave negative results for blaGES gene. This study aimed to investigate the ?-lactamase formation and detection of blaSHV and blaGES genes in K. pneumoniae and E. cloacae isolated from gingivitis diseases.

Design, Molecular Docking, Synthesis of Aromatic Amino Acids Linked to Cephalexin.

Infections caused by bacteria have a significant impact on public health. Chemical synthesis of new derivatives of cephalexin inked to amino acid (tryptophan or histidine) through an amide bond at the acyl side chain is achieved. This is a new
 approach of incorporating, tryptophan and histidine into the the primary amino group of cephalexin, in order to provide a bulky group very close to the β-lactam ring. This chemical addition act as isosteric group to the alkoximino that protect beta lactam ring from bacterial beta lactamase enzyme. The new derivatives may show resistance to β-lactamases, improve activity and pharmacokinetic properties and may give new life for old drugs that are susceptible to hydrolysis by most β-lactamases. The chemical structures of these derivatives were confirmed by: FTIR, 1H-NMR spectroscopy, elemental micro analysis and some physical properties. Molecular docking on serine beta lactamase and prediction of ADME parameters were recorded using GOLD suite and Swiss ADME software respectively. Docking scores of the new derivatives of Cephalexin on β-lactamases were higher than those of Cephalexin, which may indicate better activity

Occurrences of Metalo-β-lactamase Producing Pseudomonas aeruginosa among Clinical Samples in Kwara state, Nigeria

Pseudomonas aeruginosa is a frequent nosocomial pathogen that causes severe disease in many clinical and community settings. The objective of this paper was to isolate Pseudomonas aeruginosa from clinical samples and to investigate the occurrence of metallo β- lactamase enzyme production by collecting 145 males and 90 females’ human clinical specimens from five selected health institutions within Kwara state, Nigeria. The samples were cultured immediately using standard microbiological procedures. Multiple drug resistance patterns of the bacteria to different antibiotics were determined using the Bauer Kirby disc diffusion technique. Metallo – β lactamase production was determined using E – test strip. Data were subjected to descriptive statistics using Statistical Package for Social Sciences (SPSS) software. A total of 145 isolates were identified as Pseudomonas aeruginosa from the clinical samples.Thirty were positive for metallo β lactamase production; 11 (8 %) males and 19 (13 %) females. Absolute resistance to ceftazidime (100 %), gentamicin (100 %), ceftriaxone (100 %) were observed while low resistance to ciprofloxacin (12.4 %), piperacillin (6.9 %) and imipenem (6.9 %). All isolates were sensitive to colistin. This study had demonstrated that there is a high occurrence of metallo β lactamase enzyme producing and antibiotic-resistant strains of P. aeruginosa in clinical specimens from the studied area. Necessary measures must be implemented to stop the problems of this antibiotic resistance.

1436. Overview of ESBL Enzyme Producing Infections and UTIs

Abstract Background Extended-spectrum beta lactamase (ESBL) enzymes are plasmid-mediated, rapidly emerging and complex thereby posing a major therapeutic challenge in the management of urinary tract infections (UTIs) in community and hospital settings. In 2017, there were an estimated 197,400 cases of ESBL-producing Enterobacterales among hospitalized patients and 9,100 estimated deaths in the United States. Methods We conducted a retrospective cohort study using a publicly accessible National Inpatient Sample (NIS) database from October 2015 to December 2017. Adult patients (age >/= 18 years old) with UTI as a principal diagnosis were included. SAS 9.4 was used for univariate and multivariate linear. Logistic regression statistical analyses were used to compare mean age at the time of admission, length of stay, in-hospital mortality, hospitalization costs, and Elixhauser comorbidity indices. Results Of the total 5,776,156 patients included in the study, 52,765 patients had ESBL-enzyme induced UTIs. 66% were females and 34% were males. 63.3% were Caucasian, 11.6% were African-American, 18.8% were Hispanic, and 4.4% were Asian or Pacific Islander. The most common comorbidities were renal failure (22.8%), diabetes mellitus with complications (20.8%), congestive heart failure (20.5%), chronic lung disease (20.0%), neurological diseases (17.8%), obesity (12.6%), paralysis (12.5%), and depression (11.5%). In-hospital mortality was 2.5% (p< 0.0001), which was most likely due to the underlying co-morbidities. In patients without ESBL-enzyme induced UTIs, average length of stay was 7.8±8.5 days, and economic burden was &16,166.8 ± &21,183.5 USD. In comparison, patients with ESBL-enzyme induced UTIs had in-hospital mortality of 3.9%, average length of stay of 7.0 ± 9.7 days, and economic burden of &15,793.3 ± &29,268.6 USD. ESBL and UTI data analysis image 1 ESBL and UTI data analysis image 2 Conclusion We found that ESBL-enzyme-producing UTIs have statistically significant prolonged length of stay and economic burden, though in hospital mortality rate is low. This could be due to judicious use of antimicrobial therapy. There is a need for further research, as well as increased antimicrobial stewardship for UTIs, a globally recognized major cause of nosocomial acquired infections. Disclosures All Authors: No reported disclosures

De-Novo Design of Hits Against New Delhi Metallo-β-Lactamase Enzyme

The New Delhi Metallo-β-lactamase 1 (NDM-1) is a class of Metallo-β lactamase enzyme. It is responsible for hydrolyzing almost all β-lactam antibiotics, leading to multi-drug resistance in bacteria. The lack of specific therapeutic options against this target creates an emerging need to develop new molecules against it. The multistep fragment- and knowledge-based de-novo design methods were considered for this study to design small molecules. The designed molecules were evaluated by molecular docking and dynamics simulation, followed by drug-likeness prediction. This study reports that a new drug-like chemical entity exhibits good binding behavior against the MDM-1 enzyme. Nonetheless, in-depth biological evaluation is required to determine the efficacy of the designed binders to develop new therapeutics against NDM-1.

Comparison between Phenotypic and Genotypic Detection of Metallo Beta Lactamase Enzyme among Gram Negative Bacteria Isolated from Burn Patient

Background: -lactamases (MBLs) are emerging worldwide as powerful resistance determinants in Gram negative bacteria Rapid dissemination and spread between different bacterial species by large gene transfer favor by globalization and travel represent a high risk of worldwide pandemic among Enterobacteriaceae Objective:This study design to screen for phenotypic detection of MBL and genes coding for metallo - lactamase(MBL) such as, blaVIM, blaIMP and bla NDM among infected burn wound patients in Sulaimani city /Iraq. Materials and Methods: Out of 230 burn wound samples, 201 wound swabs and 29 tissue biopsy were collected from hospitalized burn patients with second and third degrees burnfrom Burn and Plastic Surgery Hospital in Sulaimani city from the period of April to October 2011. According to direct gram stain , bacterial cultural , biochemical tests, analytic profile identification(API) system and vitek 2 Compact system , antimicrobial susceptibility Testing by using Kirby-Bauer disk diffusion method and vitek compact system, -lactamase by Double disk synergy test and Combined EDTA disk test and Modified Hodge test and -lactamase enzyme by using polymerase chain reaction. Results: Out of 230 burn wound samples177 samples gram negative bacteria were isolated; inflammatory cells showed significantly associated with positive bacterial culture. The most frequent bacteria isolated were Pseudomonas species 48(27.12%), in which Pseudomonas aeruginosa account for a higher percentage 46(25.98%) and one species (0.56%) of each P. stutzeri and P.florescence wasisolated followed by Acinetobacterspecies 44 (24.86%) in which Acinetobacterbaumanniiwas the commonest and accounts for a higher percentage 41 (23.16%) and one species(0.56%) of each of A. ursingi, A. hemolyticasandA.complexwere identified. On the other hand in the family of Enterobacteriaceae,the most common bacteria were Klebsiellapneumoniae44(24.86%), Enterobacter cloacae 18 (10.17%) and Escherichia species 11(6.21%). The susceptibility of bacterial isolates against 18 antibiotics from different classes of antibiotics was tested and it was found that most of the isolated species of non fermenter bacteria such as Acinetobacterand Pseudomonas species show multidrug resistant pattern and high resistance against most of the antibiotics commonly used. The most resistant antibiotics against non fermenter bacteria were Ticarcillin (81.82%) against Pseudomonas species and Ticarcillin, Tazobactam- Pipracillin and Cefoxitin (93.18%) against Acinetobacterspecies .The most resistant antibiotic against K. pneumonia andE.cloacaewere Cefoxitin(84.9%)and Amoxicillin Clavulanic acid (84.9%).The most resistant antibiotics against Escherichia specieswere Cefotaxime, Trimethoprim and Cefipime (90.91%) but the most effective antibiotic with a highsensitivity rate was Imipenem (90.91).Phenotypic tests were carried out for the detection of MBL enzyme for all studied isolates and it was found that Combined disk test was the most sensitive test giving the highest percentage (31.07%) followed by Double disk synergy test(28.8%), Modified Hodge test (20.9%) .Polymerase chain reaction assay was used for genotypic detection of MBL genes (blaIMP,blaVIM, ,blaNDM ) in all isolates and the results revealed that the gene blaIMPwas locatedin 33(18.64%),blaVIM,in19 (10.73%), 2(1.12%)forblaNDM Also these genes were detected in 25 stock cultures of Gram negative bacteria preserved in 25 °C since 2008 to 2010. blaIMP gene was detected in11(44%) , blaVIM in 12(48%) and 2(8%) for blaNDM, all these genes were detected in K. pneumonae,P. aeruginosa and A. baumanniiwith length amplified genes (230) bp for IMP and (390)bp for VIM and( 621)bp for NDM. Conclusions:These results indicate that most of the Meropenem resistant strains from infected burn wound strains in this study were producing MBL enzymes The presence of MBL genes among Meropenem sensitive strains indicates that there might be a hidden MBL gene among isolated strains which cannot be diagnosed by phenotypic tests leading to the dissemination of these genes in the hospital silently among patients even within normal health workers who act as carriers for MBL genes in future

Outcome of urinary tract infection caused by Extended Spectrum Beta-Lactamase (ESBL) producing Escherichia coli and Klebsiella pneumoniae in Dr Zainoel Abidin General Hospital Aceh

Introduction : Extended-spectrum ?-lactamase (ESBL) is a mutated ?-lactamase enzyme that can hydrolyze beta-lactam antibiotics. The aim of this study was to determine the incidence rate, characteristics of patients, antibiotic use and outcome of urinary tract infections (UTIs) due to ESBL-producing Escherichia coli and Klebsiella pneumoniae at Dr Zainoel Abidin General Hospital Aceh.Method : The samples of this study were all Escherichia coli and Klebsiella pneumoniae isolated from urine culture of UTI cases. Patient characteristics and antibiotic use data were extracted from medical records. UTIs outcome was assessed as clinical improvement or death within a maximum of 30 days of treatment.Result : The results of this study obtained 63 patients with UTIs caused by E. coli and K. pneumonia of which 52.4% of them were ESBL producers. The incidence of UTIs due to E. coli was higher than that of K. pneumoniae, 63.5% and 36.5% respectively. E. coli ESBL producers were more in number than non-ESBL, conversely K. pneumoniae were mostly non-ESBL. The characteristics of patients with UTIs caused by E. coli and K. pneumoniae were predominant women 52.4%, and most cases were at 56-64 years old. Antibiotic therapy that given before and after culture results to UTI patients were generally ceftriaxone. UTIs outcome due to ESBL producing Escherichia coli and Klebsiella pneumoniae showed that 26/33 (78.8%) patients experienced improvement, however, 7/33 (21.2%) patients died.Conclusion : The irrational use of ceftriaxone in patients with UTI caused by ESBL producing E. coli and K. pneumoniae has led to a poor outcome for the patient.

Prevalensi Gen OXA-23 pada Isolat Klinis Bakter Acinetobacter baumanii di RSUP Dr. M. Djamil Padang

Acinetobacter baumannii is such a big challenge in this modern era because of their multi-drug resistance (MDR) through their resistance mechanism, namely the production of beta lactamase enzymes, one of them is oxacillinases. Several studies have revealed that the OXA-23 gene has an important role in the production of the beta lactamase enzyme so that these bacteria are resistant to antibiotics, especially carbapenem. This study was conducted to determine the description of the resistance of the Acinetobacter baumannii by looking at the prevalence of the OXA-23 gene in clinical isolates of these bacteria at RSUP Dr. M. Djamil Padang.
 This study was a descriptive observational study and used a cross-sectional design using polymerase chain reactions (PCR) and electrophoresis techniques. The samples were clinical isolates of Acinetobacter baumannii bacteria stored in the Microbiology Laboratory of the Faculty of Medicine, Andalas University with inclusion and exclusion criteria.
 Of the 150 clinical isolates of Acinetobacter baumannii studied, most of the bacteria were still sensitive to the antibiotics of Amikacin (84%), Tigecycline (78.76%), and Trimethoprim / Sulfamethoxazole (76.77%). Meanwhile, all of the Acinetobacter baumannii isolates were resistant to cefazolin. In PCR and electrophoresis tests, 50 clinical isolates were positive for the OXA-23 gene. 46 of them were carbapenem resistant, while the other 4 isolates were sensitive to carbapenem. From these results it can be concluded that the prevalence of the OXA-23 gene in Acinetobacter baumannii isolates in RSUP Dr. M. Djamil Padang was 85.18% for carbapenem resistant isolates and 4.30% for carbapenem sensitive isolates.
 Keywords : Acinetobacter baumannii, carbapenem, OXA-23 gene

Antibiotic sensitivity on pathogenic bacteria causing bacterial vaginosis

Objectives: To identify the sensitivity of antibiotics to pathogenic bacteria that cause Bacterial Vaginosis (BV).Materials and Methods: This type of research was an observational study with a sample of six specimens. The data were taken using primary data from patients who were swabbed in the vagina and then diagnosed BV with amsel criteria on vaginal secretion specimens carried out at Tanggul health center on January 23-February 23, 2020. The specimens were sent to Parahita Clinical Laboratory for bacterial identification and adjusted for sensitivity with CLSI using vitek 2 compact tool.Results: The results of this study identified the bacteria that caused bacterial vaginosis, the E. coli and K. pneumoniae with one sample of suspected ESBL. ESBL is a beta lactamase enzyme produced by bacteria and can induce bacterial resistance to penicillin, cephalosporin generation 1, 2, and 3. The types of bacteria found were E. coli and K. pneumoniae with high sensitivity antibiotics tested including piperacillin/tazobactam, ceftazidime, cefepime, ertapenem, meropenem, amikacin, gentamicin, tigecycline, and nitrofurantoin. Antibiotics with high levels of resistance tested against these bacteria included: ampicillin, amoxicillin, and ampicillin/sulbactam due to the mechanism of beta-lactam antibiotic resistance in the production of beta lactamase from bacteria.Conclusion: The type of bacteria found was E. coli and K. pneumoniae with high resistance levels in beta lactam antibiotics.

PREVALENCE AND ANTIMICROBIAL SUSCEPTIBILITY PATTERN OF EXTENDED SPECTRUM ?-LACTAMASE (ESBL) PRODUCING GRAM NEGATIVE BACILLI (KLEBSIELLA PNEUMONIA) IN A TERTIARY CARE TEACHING HOSPITAL, IN SOUTHERN-WEST KOTA DISTRICT OF RAJASTHAN (INDIA).

Background and Objectives: The resistance to broad spectrum ?-lactams which is mediated by the extended spectrum beta lactamase (ESBL) enzyme is an increasing problem now-a-days. This resistance mechanism has been responsible for nosocomial outbreaks, serious therapeutic failure if it is not detected on time and the outbreak of multidrug resistant, gram negative pathogens that need expensive control measures. Aim: To know the prevalence of ESBL production in klebsiella pneumoniae and antibiotic sensitivity pattern to 3rd generation of cephalosporins in Govt. Medical College Kota. Material and Methods: The study was conducted in Department of Microbiology, Kota Medical College, Kota, Rajasthan, from dec.2018 to dec.2019. Due permission from institutional ethical committee was obtained. This is descriptive observational study. All data were collected and analyzed with the help of suitable statistical parameters. A total of 101 consecutive, nonrepetitive, gram negative isolates, which were resistant to one of the third generation cephalosporins (cefotaxime,ceftriaxone or ceftazidime) were selected as “Suspicious for ESBL production” as recommended by the Clinical and Laboratory Standards Institute (CLSI). These isolates were confirmed for ESBL production by the double disc synergy test (DDST) and the phenotypic confirmatory disc diffusion test (PCDDT) and they were further confirmed by the E-test ESBL strip randomly. Result: Out of the 101 isolates by double disc synergy test (DDST) 45.54% (46) of Klebsiella pneumoniae isolates were ESBL producers, while PCDDT detected 58.41% (59) Klebsiella pneumoniae isolates as ESBL producers (p value<0.05). Randomly selected isolates were further confirmed by the E-test ESBL strip, which showed a highly significant correlation with PCDDT. Key words: Antimicrobial Susceptibility, Klebsiella pneumoniae, Gram Negative bacilli, Cephalosporins

Detection of Gram Negative Bacilli Producing Extended Spectrum Beta Lactamase on Intensive Care Unit at Tertiary Care Hospital

Antimicrobial resistance is a budding threat worldwide. The every class of antibiotic agents must have resistance mechanisms.The principal mechanism for resistance to the β-lactam antibiotics in gram-negative bacteria is the production of β-lactamase. The creation of extended-spectrum β-lactamases (ESBLs) is a vital mechanism which is responsible for the resistance to the cephalosporins. During the last 2 decades, ESBL producing gram-negative bacilli have arose as a major problem in many settings. Resistance to 3rd generation cephalosporins by attainment and manifestation of extended spectrum beta lactamase (ESBL) enzymes among gram-negative bacilli is on a rise. To isolate the ESBL strains from various clinical samples in ICU. To find out the prevalence of ESBL producing gram negative bacilli during the period of December 2017 - December 2018 in the Intensive care unit of Sree Balaji Medical College and Hospital. Totally 27 out of 139 gram negative bacilli (19.42%) were found to be ESBL producers. ESBL triggering gram negative bacilli spiteful the biological sample like blood, urine, wound swab, sputum were 36.36%, 16.00%, 10.00%, 9.09% individually Though Meropenem is 100% sensitive to all ESBL beginning gram negative bacilli, but still sensitivity also witnessed with some cheaper drugs like Cotrimoxazole (33.33%), Amikacin (48.14%), Gentamicin (40.7%), Ciprofloxacin (22.22%). Hence we care and will provide an analysis and treatment for affected patients with Extended Spectrum Beta Lactamase producing organisms.

Detection Of β-lactamases Enzymes Responsible For β-lactam Resistance In Some Gram Negative Bacteria By PCR

Abstract: 
 Objective(s): β- lactamases enzymes are well studied in literature since they lead to β-lactam resistance among many pathogens, hence global concern of human and animal health issues is rising. The aim of study is to detect these kinds of enzymes among some gram negative bacteria isolated locally.
 Material and methods: 30 isolate of each gram negatives Citrobacter spp., Serratia spp., and Enterobacter spp. were obtained from different clinical samples in Baghdad and Al-Najaf cities through March 2020 until January 2021. Isolates were identified biochemically and by Vitek2 system. β- Lactamases encoding genes like (blaSHV (231bp), blaIMP-1 (500), blaVIM (382), blaAmp (1150), blaCMY (1014), bla oxa23 (501), and bla oxa51 (353bp) were detected by PCR.
 Results: It was found that (80%, 75%, 30%, 50%, 38%, 55%, and 69%) of Citrobacter spp isolates had β- Lactamases encoding genes (blaSHV, blaIMP-1, blaVIM, blaAmp, blaCMY, bla oxa23, and bla oxa51), respectively. Serratia spp. showed having (blaSHV, blaIMP-1, blaVIM, blaAmp, blaCMY, bla oxa23, and bla oxa51) among (20%, 85%, 60%, 28%, 40%, 67%, and 58%), respectively. There were also (60%, 50%, 25%, 33%, 60%, 81%, and 73%) of Enterobacter spp. isolates harbored (blaSHV, blaIMP-1, blaVIM, blaAmp, blaCMY, bla oxa23, and bla oxa51), respectively.
 Conclusion: Our findings revealed that most of gram negative bacteria isolated locally had different β- Lactamases encoding genes, leading to increase health problems among patients infected with those pathogens.

Extended-spectrum and Metallo-beta lactamase enzymes mediated resistance in Pseudomonas aeruginosa in clinically isolated specimens

Pseudomonas aeruginosa is one of the leading opportunistic pathogens, frequently highlighted for the production of Extended-Spectrum Beta-Lactamase (ESBL) and Metallo-beta Lactamase (MBL) enzymes. This is believed to be the primary inhabitant of soil due to its adaptive nature and can survive in aquatic and even in a toxic environment. The current study aimed to screen ESBL, MBL producing, and Multiple Drug Resistant (MDR) strains of P. aeruginosa. Clinical specimens collected from patients were screened for the presence of P. aeruginosa. After identification, all the isolates were tested for the sensitivity pattern following the Kirby-Bauer disc diffusion method. The presence of ESBL and MBL enzymes were detected following DDST and IMP-EDTA detection tests, respectively. The sample size used in the study was 1369, and colonies for P. aeruginosa were obtained from 126 (9.20%) samples of culture media. 54.76% (69/126) of the positive cases were detected in the female population, whereas 45.24% (57/126) in the male population. High-frequency rate (n=43/126) was detected in the age group ≥31 year, followed by 21-30 (n=35/126), and 11-20 (n=34/126) age group, the minimal frequency (n=14/126) was detected in age group 0-10. The sensitivity pattern showed that the majority of the isolates were resistant, but class carbapenem (imipenem, meropenem), aminoglycosides (amikacin, gentamicin, tobramycin), and other antibiotics like sulbactam- Cefoperazone, tazobactam-piperacillin, tigecycline, and Fosfomycin exhibited the best efficacy against P. aeruginosa. It is concluded that the emergence of resistance due to ESBL and MBL enzymes in P. aeruginosa is directly linked with a public health concern because these strains are almost resistant to a wide range of antibiotics, and only limited antibiotics are potent against these organisms.

Antibacterial Activity of Eucalyptus Camaldulensis Extracts on Staphylococcus aureus Isolated from Nasal Cavity

To study the effect of prepared Eucalyptus camaldulensis in aqueous, ethanol and flavonoid extracts byconducting susceptibility tests against Staphylococcus aureus isolated from nasal carrier of healthy subjects,in addition to detection some of virulence factors by conducting ?-lactamase enzyme test, and biofilmformation test. This study included 250 healthy subjects, the males were 125(50%) and 125(50%) females.Two swab samples were collected from each subject, the studied samples collected during one year periodfrom (April / 2018- April 2019). Different culture media and biochemical tests were used for isolationand identification the isolates as Staphyllococcus aureus, in addition to prepared three types of Eucalyptuscamaldulensis extracts in form of aqueous, ethanol and flavonoid. Susceptibility test was conduct by discdiffusion method, then study some of virulence factors of Staphyllococcus aureus by conducting ?-lactamaseenzyme test and biofilm formation test. Among 250 healthy subjects involved in this, study Staphyllococcusaureus 68 (27%) isolates were detected, isolates from male was 40 (59%), while from female was 28 (41%),only 39 (57 %) isolates were sensitive to different concentrations of Eucalyptus camaldulensis, of bothaqueous and ethanol extracts, ethanol extract had a higher effect on Staphyllococcus aureus than aqueousextract, while 45 (66%) isolates were sensitive to flavonoid extract. Twelve (18%) isolates form a biofilm intube method, and 48 (71%) isolates showed ?-lactamase test positive. A higher nasal carriage were in malesin this study than females. The anti Staphyllococcus aureus effect of Eucalyptus camaldulensis extractswere flavonoid followed by ethanol then aqueous extracts, some of Staphyllococcus aureus isolates werehighly virulence by showing a positive result for ?- lactamase enzyme and biofilm formation test. this studyshown that the flavonoid extract of E. camaldulensis have a great potential as antimicrobial agents on S.aureus than crude aqueous and ethanol extracts, many nasal pharmaceutical preparation can prepare fromE. camaldulensis in crude and flavonoid form for treatment infections and reduce nasal carrier of S. aureus.

CLAINICO-BACTERIOLOGICAL SPECTRUMAND ANTIMICROBIAL RESISTANCE PROFILEIN BACTEREMIA COINFECTING COVID-19 PATIENTS IN A TERTIARY CARE HOSPITAL IN TRIPURA

Background- Secondary bacterial infections including bacteremia have been implicated as a major cause of morbidity and mortality in COVID-19 patients So, it is important to determine the bacteriological profile of organisms responsible for bacteremia among COVID-19 patients along with their antimicrobial resistance pattern Objectives- To determine the bacteriological profile, antimicrobial resistance pattern and clinical outcome among COVID-19 patients with bacteremia as manifestation of secondary bacterial infection. Study design- Single center, cross sectional study Methods- Blood cultures were obtained from COVID-19 patients with features of bacteremia and sepsis based on Sepsis-3 criteria and serum procalcitonin level. Identification and AST were performed and the patients were followed until final outcome or discharge from hospital. Results- Among 43 blood samples obtained. 8 were positive in culture for pathogenic bacteria (18.6%). Mean age of the patients were 53.4 ± 14.9 years with male preponderance (62.5%). Mean procalcitonin level was 6.5±5.7 ng/ml. Positive history of contact was the major risk factor (62.5%) and mean duration of hospital stay was 11.1± 3.9 days. Mortality rate was 37.5%. Gram negative bacilli were the major isolates (75%) and one case was caused by an unusual organism Erysipelothrixrhusiopathiae. Among the Gram negative bacilli, maximum resistance was against Amikacin (100%) and minimum against Meropenem (16.7%). Out of GNB isolates, one isolate of Pseudomonas aeruginosa was Metallo ? lactamase enzyme producer and another one was multidrug resistant strain. Conclusion- Bacteremia among COVID-19 patients is a serious form of bacterial co-infection, leading to increased morbidity and mortality among the patients. High degree of clinical suspicion and increasing prolactin level must be complemented with blood culture to rule out bacteremia in these patients, where timely intervention and proper antimicrobial therapy can be lifesaving. Keywords- COVID-19, SARS-CoV-2, Coronavirus, Bacterial coinfection, Bacteremia, Antimicrobial resistance.

Rapid detection of Klebsiella pneumoniae producing extended spectrum β lactamase enzymes by infrared microspectroscopy and machine learning algorithms.

Antimicrobial drugs have played an indispensable role in decreasing morbidity and mortality associated with infectious diseases. However, the resistance of bacteria to a broad spectrum of commonly-used antibiotics has grown to the point of being a global health-care problem. One of the most important classes of multi-drug resistant bacteria is Extended Spectrum Beta-Lactamase-producing (ESBL+) bacteria. This increase in bacterial resistance to antibiotics is mainly due to the long time (about 48 h) that it takes to obtain lab results of detecting ESBL-producing bacteria. Thus, rapid detection of ESBL+ bacteria is highly important for efficient treatment of bacterial infections. In this study, we evaluated the potential of infrared microspectroscopy in tandem with machine learning algorithms for rapid detection of ESBL-producing Klebsiella pneumoniae (K. pneumoniae) obtained from samples of patients with urinary tract infections. 285 ESBL+ and 365 ESBL-K. pneumoniae samples, gathered from cultured colonies, were examined. Our results show that it is possible to determine that K. pneumoniae is ESBL+ with ∼89% accuracy, ∼88% sensitivity and ∼89% specificity, in a time span of ∼20 minutes following the initial culture.

Detection of metallo-beta lactamases and oxacillinase genes in carbapenem-resistant Acinetobacter baumannii strains isolated in Morocco

Introductionresistance to carbapenem is widespread among Acinetobacter baumannii (A. baumannii) strains. Metallo-beta lactamases enzymes (MBL) are responsible for carbapenem resistance, as are oxacillinases (OXA). In recent years, MBL producing carbapenem-resistant strains have been reported in the world and Morocco at increasing rates. Our study aimed to investigate the presence of carbapenemases in acinetobacter strains isolated from hospitalized patients in CHU Fez.Methodsa total of 58 imipenem-resistant A. baumannii strains isolated from clinical samples were investigated. The presence of MBL was described phenotypically by the double-disk synergy test (DDST), MBL E-test, and modified Hodge test. The blaIMP, blaVIM, genes, and blaOXA-23, blaOXA-51 genes were investigated by multiplex polymerase chain reaction (PCR). The blaNDM-1 gene was determined by simplex PCR.Resultsfifty-eight strains were resistant to imipenem (98%), the modified Hodge test (MHT) was positive for 58 strains (100%), 47 strains (82%) were found to be positive for MBL by the test of double-disk synergy (DDST), 58 strains (100%) were positive by E-test MBL. The OXA 51 gene was detected in all strains, and the OXA 23 gene was detected in 53 strains (91%). In addition, the MBL genes were not detected by genotypic methods.Conclusionthe OXA-23 and OXA-51 carbapenemases type are responsible for the resistance to carbapenems in A. baumannii resistant to carbapenems in our establishment. Resistance to carbapenems by MBL enzymes has been found by phenotypic tests, which must be confirmed by genotypic methods; and solicit other MBL genes.

Detection of blaCTX-M Genes among Extended Spectrum Beta Lactamase producing Pseudomonas aeruginosa isolated from Clinical Specimens in Erbil

Pseudomonas aeruginosa is one of the most common opportunistic pathogens that cause infections. The drug resistance in Pseudomonas aeruginosa may relate to the production of broad spectrum beta lactamase enzymes. This study aimed to detect multidrug resistant, extensively drug resistant and pan drug resistant isolates of Pseudomonas aeruginosa and to determine the frequency of extended spectrum beta lactamases enzymes by using a reliable test for detection blaCTX-M1, blaCTX-M2 and blaCTX-M3 genes in Pseudomonas aeruginosa. Pseudomonas aeruginosa isolated from different clinical samples, including sputum, wound, burn and urine. Identification and antibiotic susceptibility test were made by VITEK® 2 automated system. Identification confirmed by 16S ribosomal ribonucleic acid. The bacterial deoxyribonucleic acid was extracted and specific primers were used for detection blaCTX-M1, blaCTX-M2 and blaCTX-M3 genes. 62 Pseudomonas aeruginosa strains were obtained from 650 different clinical specimens (9.54 %). Of the 62 Pseudomonas aeruginosa isolates, the highest resistances were related to the antibiotics of ticarcillin, ticarcillin/clavulanic acid, piperacillin, piperacillin/tazobactam, ceftazidime and cefepime (100 %). However, the lowest resistance was observed to meropenem, imipenem (3.23 %), amikacin, ciprofloxacin (6.45 %) and gentamycin, tobramycin (19.35 %). Out of 62 bacterial isolates, 46 (74.19 %) were multidrugresistance strains, 2 (3.23 %) were extensive drug-resistance and no pan drug resistance was isolated. The gene frequency of blaCTX-M1 was 12 (19.35 %) and blaCTX-M2 5 (8.06 %), all specimens were negative for blaCTX-3 gene. This study highlights the increased prevalence of multidrug resistant Pseudomonas aeruginosa. The blaCTX-M genes provide Pseudomonas aeruginosa with an additional powerful resistance mechanism with potentially serious clinical implications, including limitation of the therapeutic options. The incidence of extended spectrum beta lactamases varies with geographic location and time.