Isolates Of Klebsiella Spp Research Articles

Antibacterial Susceptibility Pattern of Bacteria Isolated from Ready-to-Eat Lettuce and Gurasa Sold within Kaduna State University (Main Campus), Kaduna State

Study’s Novelty/Excerpt This study evaluates the antibacterial susceptibility of pathogens isolated from ready-to-eat lettuce and gurasa sold at Kaduna State University, identifying significant health risks associated with these popular foods. The research reveals a high prevalence of antibiotic-resistant bacteria, including Staphylococcus aureus, Escherichia coli, Bacillus spp, Salmonella spp, and Klebsiella spp, with S. aureus and E. coli being the most common isolates. The findings highlight the urgent need for improved food safety measures and antibiotic resistance management to mitigate potential public health threats. Full Abstract The consumption of ready-to-eat lettuce and gurasa has gained acceptance due to its appealing taste and nutritional value. However, these foods can serve as a vehicle for transmission of pathogens. This study was carried out to assess the antibacterial susceptibility pattern of pathogens isolated from lettuce and gurasa sold at Kaduna State University. Samples were collected from five vendors of gurasa and lettuce. Standard microbiological methods were carried out using a pour plate and a modified Kirby-Bauer disc diffusion method to determine the antibacterial susceptibility pattern. The isolates obtained were examined for morphological and biochemical characteristics. Escherichia coli, Bacillus spp, Salmonella spp, Staphylococcus aureus, and Klebsiella spp. were isolated and identified. Staphylococcus aureus was the most prevalent bacterium in this study, having 27.7% in lettuce and 38.4% in gurasa, and Escherichia coli, having 16.6% in lettuce and 30.7% in gurasa. There was no significant difference (P ˃ 0.05) in the total colony counts of bacteria among the samples. Lettuce had the least bacteria count (1.48 x 106 CFU/g), while gurasa had the highest (1.55 x 106 CFU/g). Antimicrobial sensitivity test results showed that 10 S. aureus isolates were resistant to Rocephin (100%), 7 E. coli isolates were resistant to Septrin, Amoxicillin, and Augmentin (100%), 6 Klebsiella spp isolates were resistant to Amoxicillin and Augmentin(100%), 5 Salmonella spp isolates were resistant to Septrin, Amoxicillin and Augmentin (100%) and 3 Bacillus spp were resistant to Rocephin (100%). The high bacterial resistance to antibiotics is of great concern as infections with these organisms could be lethal.

Phenotypic detection of extended spectrum beta-lactamase production in clinical and non - clinical isolates of klebsiella species of animal origins in Abeokuta, Nigeria.

Production of extended-spectrum β-lactamases (ESBLs) confer resistance to β-lactam antibiotics. This study investigated the presence of ESBL-producing Klebsiella spp in clinical and non-clinical samples from different animal species in Abeokuta, Nigeria. The species of Klebsiella were determined by biochemical characterization (Oxoid Microbact GNB 24E®), while phenotypic ESBL-production was confirmed by using the cefpodoxime and cefpodoxime/clavulanic acid combination disc kit. Ninety-five Klebsiella isolates were investigated in this study. Fifty-five (57.9%) of the isolates were obtained from the faeces of apparently healthy animals while 40 (42.1%) were from clinical samples. The Klebsiella isolates were identified as follows: K. oxytoca (34.7%), K. pneumoniae (26.3%), K. ozaenae (18.9%), K. terrigena (13.7%), K. rhinooscleromatis (5.3%), K. planticola (1.1%). Eight (8.4%) out of the 95 isolates of Klebsiella spp were identified as ESBL-producers. These included four isolates of Klebsiella pneumoniae, three of Klebsiella oxytoca and one of Klebsiella ozaenae. Five out of the eight ESBL-producing isolates were from clinical samples while three isolates were from the faeces of apparently healthy animals. The ESBL-producing isolates were 100% resistant to ampicillin, cefotaxime, ceftazidime, cefpodoxime, streptpmycin, tetracycline, compound sulfonamide, trimethoprim/sulphomethoxazole, and nalidixic acid. Apparently healthy carriers and sick animals can serve as sources of transmission of ESBL-producing Klebsiella spp to other animals and humans.

Frequency Of Uropathogens Showing Resistance To Nitrofurantoin.

Urinary tract infection (UTI) caused by bacteria is the commonest infection accountable for the unforeseen healthcare cost throughout the globe. Nitrofurantoin is being studied as a solution to the perpetually increasing threat of antimicrobial resistance. The objectives of this study were to determine the frequency of urinary isolates causing UTI and their susceptibility pattern against Nitrofurantoin. Data of all isolates reported as uropathogens from April 1, to December 31, 2021, was collected through Electronic Medical Record system of Shalamar Hospital, Lahore. Results of Nitrofurantoin susceptibility were recorded to find the resistance pattern of bacterial isolates. Out of a total of 3,221 samples, 672 (20.9%) were positive with significant bacteriuria. Of the positive samples, 418 (62.2%) were collected from females and 254 (37.8%) from males, with female to male ratio of 1.65:1. The number of female patients was higher in adult age. Of the positive samples, E. coli was the commonest isolate seen in 390 (58%) of samples, followed by Enterococcus spp. 92 (13.7%), Klebsiella spp. 86 (12.8%), Pseudomonas spp. 35 (5.2%), Staphylococcus saprophyticus 24 (3.6%), Proteus spp. 21 (3.1%), Citrobacter spp. 15 (2.2%), and Acinetobacter spp. 9 (1.3%). Overall, 587 (87.4%) isolates were sensitive to Nitrofurantoin. However, it showed increased resistance to 28 (32.6%) isolates of Klebsiella spp. E. coli remains the commonest uropathogen. In conclusion, Nitrofurantoin can be used to treat UTI caused by common bacterial pathogens except Klebsiella spp.

Plasmid-mediated quinolone resistance genes detected in Ciprofloxacin non-susceptible Escherichia coli and Klebsiella isolated from children under five years at hospital discharge, Kenya

BackgroundThe increasing spread of fluoroquinolone resistant enteric bacteria is a global public health concern. Children recently discharged from the hospital are at high risk of carriage of antimicrobial resistance (AMR) due to frequent exposure to antimicrobials during inpatient stays. This study aimed to determine the prevalence, correlates of ciprofloxacin (CIP) non-susceptibility, and distribution of plasmid-mediated quinolone resistance (PMQR) genes in Escherichia coli (E. coli) and Klebsiella spp isolated from children under five years being discharged from two Kenyan Hospitals.MethodsE. coli and Klebsiella spp were isolated from fecal samples from children discharged from hospital and subjected to antimicrobial susceptibility testing (AST) by disc diffusion and E-test. CIP non-susceptible isolates were screened for seven PMQR genes using multiplex polymerase chain reaction (PCR). Poisson regression was used to determine the association between the carriage of CIP non-susceptible isolates and patient characteristics.ResultsOf the 280 CIP non-susceptible isolates: 188 E. coli and 92 Klebsiella spp isolates identified among 266 discharged children, 195 (68%) were CIP-non-susceptible with minimum inhibitory concentrations (MICs) of ≥ 1 µg/mL. Among these 195 isolates, 130 (67%) had high-level CIP MIC = ≥ 32 µg/mL). Over 80% of the isolates had at least one PMQR gene identified: aac(6’)lb-cr (60%), qnrB (24%), oqxAB (22%), qnrS (16%), and qepA (6%), however, qnrA was not identified in any isolates tested. Co-carriage of qnrB with acc(6’)-lb-cr was the most predominant accounting for 20% of all the isolates. Ceftriaxone use during hospital admission and the presence of extended spectrum beta-lactamase (ESBL) production were significantly associated with the carriage of CIP non-susceptible E. coli and Klebsiella spp.ConclusionCIP non-susceptibility is common among E. coli and Klebsiella spp isolated from hospital discharged children in Kenya. Carriage and co-carriage of PMQR, including the newly identified qepA gene, were frequently observed. These findings suggest that children leaving the hospital may serve as an important reservoir for transmission of resistant E. coli and Klebsiella spp to the community. Enhanced surveillance for AMR determinants is critical to inform interventions to control antimicrobial-resistant bacteria.

Analysis of Extended Spectrum Beta Lactamase Frequency in Klebsiella spp Isolates

The issue of increasing resistance to antibiotics in recent years has become an important problem all over the world. Our aim is to determine the antimicrobial resistance profile and Extended Spectrum Beta-Lactamase (ESBL) rates in Klebsiella spp isolates to prevent the gradual increase in multi-resistant isolates as a result of unconscious antibiotic use thereby contributing to the faster effective treatment of infections. A total of 100 Klebsiella spp were isolated and identified from various clinical specimens. Antibiotic susceptibility tests were performed using the Kirby-Bauer method. The presence of extended-spectrum beta-lactamases (ESBL) was detected using the Double Disc Synergy Test (DDST) and E-test methods. The rates of ESBL-producing strains were 46.1% in 6 K. oxytoca and 56.3% in 49 K. pneumoniae. These strains were found to be 38% in 38 adult patients and 17% in 17 pediatric patients, and this difference was statistically significant (p <0.05). The ESBL rate was 31% in 31 male patients and 24% in 24 female patients, and this difference was not statistically significant (p>0.05). This rate was found to be high in patients hospitalized in the pediatric service and intensive care unit. 67 out of 100 strains were found to be suspicious for ESBL by Disk Diffusion Test (DDT). DDST and E-tests were applied as confirmatory tests. The sensitivity of the DDST and E tests was 100%. Screening for ESBL in Klebsiella spp and other members of Enterobacteriaceae isolates is necessary to reduce further selection and spread of these increasingly broad-spectrum antimicrobial-resistant enteric pathogens.

Bacteriological study and its antibiotics susceptibility pattern of Otitis Media in Iraqi patients

Otitis media is an acute upper respiratory tract infection-related inflammation of the middle ear and tympanic membrane, frequently affecting children. Typically, a subsequent bacterial infection complicates a viral infection, which ultimately causes the condition. The study aims to study the function of bacterial ear infections and its causes, as well as their resistance to medications, which was the focus of this investigation. The first axis of the research was the identification of bacterial isolates using recognized diagnostic tools, and the second axis was determining the antibiotic's resistance and sensitivity. Patients with otitis media were gathered from Al-Hakim General Hospital and Al-Sadr city hospital in Al-Najaf city between November 2020 and April 2021 for 100 clinical samples. More than 80 samples were found to be infected with bacteria. Bacterial strains found in this investigation are ( 30 ) isolates of Pseudomonas aeruginosa, (20) isolates of Klebsiella spp, (20) isolates of Proteus spp, ( 15 ) isolates of Staphylococcus aureus, (8) isolates Escherichia coli and (7) isolates Enterococcus fecalies. As part of this research, the disk diffusion method was used to assess how sensitive the test was. The results showed that Pseudomonas aeruginosa was resistant to most antibiotics, particularly the penicillin family, cephalosporin, and trimethoprim, with the existence of isolates resistant to meropenem. The investigation results varied for the quinolone, aminoglycoside, and macrolide families. Klebsiella spp. were tested for antibiotic sensitivity and found to be resistant to most antibiotics, particularly those in the penicillin family, cephalosporins, and trimethoprim. Some quinolones, aminoglycosides, and macrolides are also resistant. Proteus spp were resistant to most antibiotics, particularly the penicillin family (except for augmentin, which had some sensitive isolates) and cephalosporin (except for cefdinir and cefepime) had some susceptible isolates) and trimethoprim, in addition to the presence of isolates resistant to meropenem. There is a discrepancy in the examination results for the quinolone family. The aminoglycoside family is also highly resistant. S. aureus isolates were resistant to penicillin (except for augmentin, which some isolates were responsive to), trimethoprim, and quinolones, with the presence of isolates resistant to vancomycin. The macrolide class ( azithromycin) also has a significant resistance level. Escherichia coli is susceptible to meropenem, imipenem, and certain cephalosporin generations. Augmentin, cefepime, cephalothin, meropenem, imipenem, and azithromycin were ineffective against Enterococcus fecal. The conclusion is that Pseudomonas spp has a role in ear infections and the germs Klebsiella spp., Proteus spp., Staphylococcus aureus, Escherichia coli, and Enterococcus fecalies. Penicillin and cephalosporin resistance was seen in the majority of the identified isolates. The existence of isolates of Proteus and Pseudomonas species resistant to meropenem. Vancomycin-resistant strains of Staphylococcus aureus isolates are present. Keywords: Otitis media, Resistance antibiotic, S.aureus, P.aerginosa

Antimicrobial properties and acute toxicity evaluation of Pycnanthus angolensis stem bark

The antimicrobial activity of Pycnanthus angolensis (Welw) (stem bark) used in ethnomedicine to treat urinary tract infections, diarrhoea, malaria and typhoid fever was investigated. Clinical isolates of Klebsiella spp, Proteus mirabilis, and type isolates including Candida albicans ATCC 7596, Escherichia coli ATCC 11,775, Pseudomonas aeruginosa ATCC 10,145, Staphylococcus aureus ATCC 1200 and Salmonella kintambo SSRL113 were used in the study. Agar well diffusion and broth dilution techniques were used to determine the antimicrobial activity and minimum inhibitory concentration (MIC), respectively. The results showed that the plant's ethanol extract was more active than the aqueous plant extract. Fractionation of the ethanol extract with n-hexane, chloroform, ethyl-acetate, acetone and methanol was carried out to obtain corresponding fractions. Antimicrobial activity testing of the fractions showed that the ethyl-acetate fraction exhibited the highest antimicrobial effect. Phytochemical analysis of the crude extracts and the various fractions revealed the varied contents of alkaloids, saponins, essential oils, tannins, glycosides and flavonoids, which are or can be largely responsible for the recorded antimicrobial effect. Acute toxicity studies performed on the ethanolic extract using albino rats indicated that the extract is non-toxic (LD50>5000 mg/kg body weight).

Pregnancy and asymptomatic bacteriuria- A study from a tertiary care teaching hospital, Hyderabad

Introduction: Asymptomatic bacteriuria is found in 2 to 10% of pregnant women. Complications like acute pyelonephritis, postpartum UTI, hypertension, anemia, prematurity, low birth weight babies and prenatal death may occur if untreated. Hence early detection and treatment are of considerable importance. Materials and Methods: A crosssectional study was conducted for a period of one year. A clean catch midstream urine sample was collected from pregnant women as a part of routine antenatal screening. The samples were processed according to standard microbiological techniques –wet mount examination, gram staining, glucose test paper, culture and antibiotic susceptibility testing. Results: Of the 200 samples received from the study group, 28(14%) were culture positive. The prevalence of culture positivity was more in the third trimester (19.4%).The predominant organisms isolated were gram negative bacteria like Escherichia coli (35.7%) and Klebsiella spp (28.5%). E.coli and Klebsiella showed variable resistance to fluoroquinolones, cephalosporins, aminoglycosides, betalactam and betalactam inhibitor combination. However the isolates of E.coli were sensitive to carbapenams and polymixins. Isolates of Klebsiella spp were sensitive to Polymixins. The predictive accuracy of different screening tests with culture was calculated. The sensitivity, specificity, positive predictive value, negative predictive value of grams staining is more compared to urine wet mount examination and urine test paper method in early detection of bacteruria. Conclusion: Early detection and treatment for asymptomatic bacteriuria is of considerable importance as 40% of asymptomatic cases develop into pyelonephritis The choice of antibiotic should be based on urine culture, stage of gestation, clinical data and the characteristics of the antibiotic. Keywords: Asymptomatic bacteriuria, Antenatal women, prevalence, Glucose test paper, Urine culture, Wetmount examination, Gram staining, Perinat

Statewide surveillance of carbapenemase-producing carbapenem-resistant Escherichia coli and Klebsiella species in Washington state, October 2012-December 2017.

Carbapenem-resistant Enterobacterales (CRE) are common causes of healthcare-associated infections and are often multidrug resistant with limited therapeutic options. Additionally, CRE can spread within and between healthcare facilities, amplifying potential harms. To better understand the burden, risk factors, and source of acquisition of carbapenemase genes in clinical Escherichia coli and Klebsiella spp isolates from patients in Washington to guide prevention efforts. Multicenter prospective surveillance study. Escherichia coli and Klebsiella spp isolates meeting the Washington state CRE surveillance case definition were solicited from clinical laboratories and tested at Washington Public Health Laboratories using polymerase chain reaction (PCR) for the 5 most common carbapenemase genes: blaKPC, blaNDM, blaIMP, blaVIM, and blaOXA-48. Case patients positive by PCR were investigated by the public health department. From October 2012 through December 2017, 363 carbapenem-resistant E. coli and Klebsiella spp isolates were tested. Overall, 45 of 115 carbapenem-resistant K. pneumoniae (39%), 1 of 8 K. oxytoca (12.5%), and 28 of 239 carbapenem-resistant E. coli (11.7%) were carbapenemase positive. Of 74 carbapenemase-positive isolates, blaKPC was most common (47%), followed by blaNDM (30%), blaOXA-48 (22%), and blaIMP (1%). Although all cases had healthcare exposure, blaKPC acquisition was associated with US health care, whereas non-blaKPC acquisition was associated with international health care or travel. We report that blaKPC, the most prevalent carbapenemase in the United States, accounts for nearly half of carbapenemase cases in Washington state and that most KPC-cases are likely acquired through in-state health care.

Rapid detection of carbapenemase production in Enterobacteriaceae by different phenotypic methods

Introduction: Carbapenemase producing enterobacteriaceae (CPE) isolates have now emerged worldwide. Resistance to carbapenems is mainly due to production of beta-lactamases inactivating carbapenems (carbapenemases) included in classes A, B, C, or D of the Ambler classification. In Enterobacteriaceae carbapenemase encoding genes are often located on plasmids that contribute to a rapid spread among clinically relevant gram negative bacteria. Rapid and reliable detection of carbapenemase production is needed for therapeutic and control reasons. Aims: To compare the different phenotypic methods of carbapenemase detection namely, the modified paper strip carba NP method, the CLSI Carba NP method, the carbapenem inactivation method, and for rapid detection of CPE. Materials and methods: A total of 200 CPE from urine, pus, and blood cultures sent to the Microbiology Laboratory, R.G. Kar Medical College and Hospital (RGKMCH), were tested for the modified paper strip carba NP method, the CLSI Carba NP method, and the carbapenem inactivation method. Results and analysis: Out of 200 isolates, 108 isolates of Klebsiella spp and 92 isolates of Escherichia coli were compared for carbapenemase production by various phenotypic methods. In total, 100(93%) isolates of Klebsiella spp and 88 (95%) isolates of Escherichia coli showed positive results by paper strip and carba NP methods. A total of 96 (89%) isolates of Klebsiella spp and 84 (91%) isolates of Escherichia coli showed positive results by CLSI carba NP method. A total of 104 (96%) isolates of Klebsiella spp and 92 (100%) isolates of Escherichia coli showed positive results by CIM method. Conclusion: Rapid and accurate detection of carbapenemase producers are important for preventing their spread in health care settings. Although genotypic tests remain the gold standard but cannot practically be conducted because they are highly expensive and results are limited by the targets. The different phenotypic methods used in this study were inexpensive, rapid, highly sensitive, and specific. The modified paper strip Carba NP method in this study is a simple and rapid method compared to those performed by the CLSI method.

Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae in Effluents of Different Hospitals Sewage in Biratnagar, Nepal

Untreated water from hospitals of Biratnagar as effluent is a serious concern from health point of view. Antimicrobial resistant bacteria are a serious threat which may contaminate the drinking water and environment. With an aim of isolation and detection of Multidrug resistance (MDR) and Extended-spectrum b- lactamases (ESBL) producing Enterobacteriaceae from the effluents and sewage samples of hospitals, this study builds the importance to inquiry about the involvement of hospital liquid waste discharge in the development and distribution of antibiotics resistance in the environment bacteria. Ten hospital sewage samples were aseptically collected, processed and analyzed. Isolates were biochemically identified, and their antimicrobial activity were tested. Of the ten sewage samples analyzed, 23 bacteria isolates were isolated which contained 8 Escherichia coli (34.7%), 5 isolates each of Citrobacter spp (21.7%) and Enterobacter spp (21.7%), 3 isolates of Klebsiella spp (13%), and 1 isolate each of Shigella spp (4.3%) and Yersinia spp (4.3%). Most of the bacteria isolated were resistant to ampicillin, ceftazidime, cefpodoxime, amoxicillin/clavulanate, cefotaxime, cefoxitin, ceftriaxone, and cefuroxime. The isolates were sensitive to nitrofurantoin, azithromycin, trimethoprim/sulfamethoxazole, chloramphenicol, ciprofloxacin, ofloxacin, and gentamycin antibiotics. Out of twenty-three isolates, 16 (69.6%) were found to be multidrug resistant, 7 (30.4%) were producing extended beta lactamase, while 18 (78.3%) multi-antibiotics resistance index greater than 20%. Among the bacteria isolated; 80% of the Citrobacter; and 75% of the E. coli were found biofilm producing bacteria. Sewage treatment plant must be established in hospital for their effluents and sludge coming from the hospital.

Correlation of quinolone-resistance, qnr genes and integron carriage in multidrug-resistant community isolates of Klebsiella spp.

Plasmid-mediated quinolone resistance (PMQR) determinants and integrons have a considerable contribution to bacterial drug resistance in Gram-negative pathogens. We studied the prevalence of PMQR genes and integron carriage in multidrug-resistant community isolates of Klebsiella spp. Two hundred and fifty Klebsiella spp. isolates were collected from outpatient specimens between August 2015 and October 2017 in Yazd central Laboratory, Iran. Antibiotic susceptibility was determined against 17 antibiotics and minimum inhibitory concentration (MIC) of ciprofloxacin was measured by E-test. Polymerase chain reaction (PCR) was employed for detection of qnrA, qnrB, qnrS, aac(6')-Ib-cr, oqxAB and qepA genes. Disc diffusion results showed that 17 isolates (6.8%) were multidrug resistant (MDR), two of which were Klebsiella oxytoca and 15 were Klebsiella pneumoniae. MIC measurements revealed 11 ciprofloxacin-resistant isolates (including the two K. oxytoca), three intermediately-resistant and three ciprofloxacin-susceptible isolates. All ciprofloxacin-resistant and intermediately-resistant isolates carried at least one and up to four PMQR genes. The most prevalent PMQR gene was oqxAB (93.75%) followed by aac(6')-ib-cr (50.0%), qnrB (25.0%) and qnrS (18.75%) but qnrA and qepA were not detected. Class 1 integron was observed in 14 (82.3%) isolates including nine ciprofloxacin-resistant, two intermediately-resistant, and three susceptible isolates. Class 2 and 3 integrons were not observed. Presence of MDR, multiple PMQR determinants as well as class 1 integron in community isolates of Klebsiella spp. can be an important source of transmission of these opportunistic pathogens.

631. Laboratory Surveillance of Carbapenem-Resistant Enterobacteriaceae in an Endemic Country, Greece, 2015–2018

BackgroundGreece is facing an endemic situation with carbapenem-resistant pathogens in the hospital sector. The Central Public Health Laboratory (CPHL) is the main laboratory for the surveillance of the carbapenem resistance mechanisms, accepting resistant isolates from hospitals on a voluntary basis. The aim of the present study is to evaluate the epidemiology of carbapenem-resistant Enterobacteriaceae (CRE) isolated in Greek hospitals the period 2014–2018 and their susceptibility to other antibiotic classes.MethodsA total of 843 CRE isolates (741 Klebsiella spp., 47 Enterobacter spp., 35 Escherichia coli, and 20 others) have been submitted from 36 hospitals throughout the country (Figure 1) to the CPHL. They originated from different clinical specimens (295 urine, 157 blood, and 107 other) or surveillance cultures (185 rectal and 23 pharyngeal swabs) or unknown (n = 76). Carbapenemase genes were detected by PCR targeting blaKPC, blaNDM, blaVIM, and bla OXA-48. Resistance to aminoglycosides and fluoroquinolones was tested with the disk diffusion method according to EUCAST guidelines.ResultsThe isolates were found positive for several carbapenemase genes (Table 1). Overall, KPC-2 (36%) was the predominant carbapenemase, followed by the metalloenzymes NDM (28%) and VIM (18%) while OXA-48 ranked fourth (7%). KPC enzyme was predominant among Klebsiella spp isolates followed by NDM, whereas among Enterobacter, E.coli, and other species, the VIM metalloenzyme predominated. Simultaneous detection of two carbapenemase genes was found in 30 (4%) isolates: 14 blaKPC/blaVIM, 4 blaKPC/blaNDM, 1 blaKPC/blaOXA-48, 5 blaNDM/blaVIM, 5 blaNDM/blaOXA-48, and 1 blaVIM/blaOXA-48. In 63 isolates (7%), the carbapenem resistance was attributed to other mechanisms, mainly production of ESBL or AmpC plus porin loss. In total, 590 (70%) CRE isolates exhibited a multidrug-resistant phenotype being simultaneously resistant to aminoglycosides and fluoroquinolones.ConclusionThe CRE isolates’ diversity, regarding bacterial species, carbapenemase types and combinations and their temporal and spatial distribution mandate a more structured, continuous laboratory surveillance to monitor the ongoing carbapenem and multidrug resistance evolution and inform infection control and public health actions. Disclosures All authors: No reported disclosures

Evaluation of microorganisms isolated from blood cultures and their susceptibility profiles to antibiotics in five years period

Aim: In starting antibiotic treatment to know the distribution of infectious agents and the antibiotic resistance rates is vital especially in critically ill patients to prevent disease progression. In this study we aimed to determine the distribution and antimicrobial resistance patterns of blood culture isolates obtained in a tertiary center. Methods: The microbiological laboratory records between January 2014 and December 2018 were retrospectively reviewed. Each conventional aerobic blood culture bottle per patient with positive results was recorded with the antimicrobial susceptibility profiles of the bacteria isolated. Descriptive statistics (number, percentage, mean and median) were performed. Comparison of descriptive data between groups was performed with cross tables and chi square test. One-way ANOVA test was used to compare the antibiotic resistance rates between spp. Results: A total of 66004 blood cultures were analyzed in this cross-sectional study. Of the 3882 (21.0%) positive results, 3256 (4.9%) were identified as contamination. The most commonly isolated microorganisms were Coagulase negative staphylococcus, Escherichia coli, Klebsiella spp., Staphylococcus aureus and Acinetobacter spp. Extended spectrum β-lactamase (ESBL) positivity was determined in 236 (62.4%) isolates of E.coli, and 186 (56.8%) isolates of Klebsiella spp. Vancomycin resistance showed a significant increase in Enterococcus spp. in time. E.coli did not show any resistance to imipenem, meropenem, tigecycline or colistin. Methicillin susceptible Staphylococcus aureus (MSSA) and methicillin resistant Staphylococcus aureus (MRSA) did not show any resistance to vancomycin, teicoplanin, linezolid or daptomycin. In Enterococcus spp., tigecycline resistance was 1.9%, while daptomycin and linezolid resistance were not determined. The most effective resistance agent to Acinetobacter spp. and Pseudomonas spp. was colistin. Fungal infection was detected in 156 patients. Candida albicans and Candida parapsilosis complex were the most common isolates. Conclusion: Antibiotic resistance rates are increasing in all over the world. Rational antibiotic usage may aid the clinicians to overcome this condition. Epidemiological data is important in this regard.

The prevalence of CMY-2, OXA-48 and KPC-2 genes in clinical isolates of Klebsiella spp.

Klebsiella pneumoniae is a Gram-negative bacterium which causes several human infections. Treatment of infections related to K. pneumoniae has become problematic, because of increasing trend of extended spectrum β-lactamases producing (ESBLs) strains. The present study was aimed to detect the prevalence of ESBL-producing Klebsiella spp. and KPC-2, CMY-2 and OXA-48 β-lactamase encoding genes in clinical isolates of Klebsiella spp. isolated from hospitalized patients. In this cross-sectional study carried out from February to August 2014, 144 isolates of Klebsiella spp. were collected from different clinical specimens in hospitals in the North of Iran. Klebsiella isolates were identified using standard microbiological procedure. Antimicrobial susceptibility patterns were determined using disk diffusion method in accordance with CLSI recommendation. The presence of antibiotic resistance genes were investigated by PCR method. Among 144 Klebsiella spp., 118 isolates were identified as K. pneumoniae and 26 isolates as Klebsiella oxytoca. Antibiotic susceptibility test showed the most resistance rates were against amoxicillin (96.5%) and ampicillin (95.8%). On the other hand, the most effective antibiotic was revealed to be imipenem with resistance rate of 4.9% (Table 2). Among 144 isolated Klebsiella strains, 57 cases (39.6%) were ESBL producers. The OXA-48 and KPC-2 genes were not detected among Klebsiella spp. during the present study, but 21.1% of isolates contained CMY-2 gene. This is the first report of CMY-2 gene detection in Klebsiella spp. in Iran. The homology between CMY-2 genes identified in isolates from Northern Iran and in other countries showed the wide dispersion of this gene around the world.

Prevalence of Extended Spectrum Beta-Lactamase Producing Escherichia coli and Klebsiella species from Urinary Specimens of Children attending Friendship International Children’s Hospital

Extended-spectrum β-lactamase producing E. coli and K. pneumoniae is a serious threat to the patients. These organisms are major extended spectrum beta lactamase (ESBL) producers. The objective of this study was to determine the prevalence of Extended spectrum β- lactamase producing strains of Escherichia coli and Klebsiella spp isolates from the urine sample of children visiting International Friendship Children Hospital. During the seven months, between June 2016 to December 2016, 1018 mid-stream urine samples(MSU) were collected from patients suspected of having UTI. The samples were investigated by conventional semi-quantitative culture technique and identification of E. coli and Klebsiella spp. was done by microscopy and biochemical test. Antibiotic susceptibility test of isolates was performed by modified Kirby Bauer Disc diffusion test. ESBL screening test was done by using 3rd generation Cephalosporin and confirmation done by combination disc diffusion method. Out of total 1018 MSU samples investigated, 200(19.64%) isolates of E. coli and 28(2.7%) isolates of Klebsiella spp. making a total of 228(22.39%) were found to cause significant bacteriuria. 76(33.33%) isolates, from those causing significant bacteriuria, were Multi-drug resistant organisms. Out of 228 isolates, 54(23.68%) were ESBL producers, that includes 51(25.5%) Escherichia coli and 3(12.5%) Klebsiella pneumoniae. ESBL producers were more common in in-patient (36.17%) than out-patient (20.44%). Most of the ESBL producers were resistance to amoxicillin, followed by Cotrimoxazole and Ciprofloxacin respectively. They were highly sensitive to Imipenem, Tigecycline, Amikacin, Piperacillin-Tazobactam, and Nitrofurantoin. High prevalence of ESBL producing E. coli and Klebsiella pneumoniae was found among children. Regular and routine monitoring of ESBL producing isolates is essential.Nepal Journal of Biotechnology. Dec. 2017 Vol. 5, No. 1: 32-38

Prevalence of Extended Spectrum Beta-Lactamases among Escherichia coli and Klebsiella spp isolates in Manipal Teaching Hospital, Pokhara, Nepal

Objective: To isolate, identify and phenotypically characterize extended spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella spp in Manipal Teaching Hospital. Methods: Cross sectional study was conducted among E. coli and Klebsiella spp recovered from patients’ various samples to establish the prevalence of organisms producing ESBL in Manipal Teaching Hospital, Pokhara, Nepal between October 2011 and April 2012. ESBL production was detected by Clinical laboratory Standards Institute (CLSI) recommendations. Results: E. coli (n=285) and Klebsiella spp (n=55) were isolated from various clinical samples. The specimens were urine 255 (75%), blood 18 (5.3%), pus 48 (14.1%), sputum 17 (5.1%), and body fluid 2 (0.6%). Seventy six (22.4%) were ESBL producing organisms by phenotypic confirmatory test with double disk diffusion method. ESBL group of organisms showed 100% resistance to ampicillin and cefotaxime. All the organisms in this study were 100% sensitive to imipenem and 95.6% sensitive to cefoperazone+sulbactam combination. ESBL producing isolates showed high rate of resistance to ciprofloxacin (90.7%), ceftriaxone (89.4%), ceftazidime (89.4%), cotrimoxazole (90.4%) and norfloxacin (88.1%) as com­pared to non-ESBL group. Conclusion: ESBL producing E. coli and Klebsiella spp showed high prevalence in Nepal. Routine laboratory testing for ESBL in Nepalese hospitals is needed in order to optimize antibiotic management and reduce the risk of spread of infec­tions caused by ESBL producers. J Microbiol Infect Dis 2015;5(2): 69-75 Key words: Extended spectrum beta-lactamase, ESBL, E. coli, Klebsiella spp, Prevalence, Nepal

Biochemical and Molecular Characterization of Food Contaminating Bacteria Isolates from Food Stall Vegetables

Today the issue of food safety is global problem that gets main concern in setting public health policy. The eruption of diseases caused by food contamination occurs in places where sanitation and hygiene conditions are generally poor. Reliable identification of bacteria remains to be an important task in food microbiology. Molecular procedures are the most efficient tools of microbial characterization. The objectives of this research were: To identify food contaminating bacteria from vegetable sample taken from food stall using biochemical tests; And to conduct molecular characterization of food contaminated bacteria isolated from vegetable sample in food stall. Six different (pair of cooked and uncooked) vegetable samples were isolated from three food stalls at Solo, Indonesia. Biochemical tests for glucose, lactose, mannitol, maltose, sucrose, SIM (H 2S, Indole, and motility), Simmon's Citrate; Methyl Red and Voges Proskauer was conducted. 16S rRNA characterization and DNA sequence analysis was done. Both the biochemical and molecular characterization revealed that the dominant bacteria contaminants in the food stall vegetable samples were: 24 isolates of Klebsiella Spp , 3 isolates of Pseudomonas aeroginosa, 2 isolates of

Epidemiology and prognostic determinants of bacteraemic catheter-acquired urinary tract infection in a single institution from 1991 to 2010

To determine the epidemiology of bacteraemic Catheter-Acquired Urinary Tract Infection (CA-UTI) and to identify independent predictors of mortality. This study was part of a bloodstream infection surveillance study that prospectively collected data on consecutive patients with bacteraemia in our institution from 1991 to 2010. Factors associated with 30-day mortality were determined. CA-UTI was the confirmed source of 1007 bacteraemias. The most common microorganisms isolated were Escherichiacoli (42%), Klebsiella spp (15%), Enterococcus faecalis (12%) and Pseudomonas aeruginosa (12%). Along the 2006-2010 periods, antibiotic-resistant E. coli and Klebsiella spp isolates accounted for 49% of the bacteraemia due to CA-UTI. Shock and mortality accounted for 125 and 92 cases, respectively (12% and 9%). Factors associated with mortality were: inappropriate empirical treatment (OR: 1.86, 95% CI: 1.48-2.44), ultimately or rapidly fatal prognosis of underlying disease (OR: 2.56, 95% CI: 1.48-4.44) and shock on presentation (OR: 12.62, 95% CI: 7.61-20.95). Inappropriate empirical treatment was most frequent in cases of bacteraemia produced by antibiotic-resistant E. coli or Klebsiella spp, Enterococcus spp. and P. aeruginosa. Factors associated with the isolation of a microorganism of this type were previous antibiotic therapy and healthcare-associated bacteraemia (OR: 1.50, 95% CI: 1.16-2.14 and OR: 3.03, 95% CI: 2.22-4.01, respectively). In cases of previous antibiotic therapy or healthcare-associated bacteraemic CA-UTI may indicate the need to initiate empirical therapy activity against antibiotic-resistant Enterobacteriaceae, E. faecalis and P. aeruginosa.

Antimicrobial resistance among producers and non-producers of extended spectrum beta-lactamases in urinary isolates at a tertiary Hospital in Tanzania

BackgroundPublished data on the existence and magnitude of extended spectrum beta-lactamase (ESBL) production in urinary pathogens in local setting is limited. The aim of the present study was to determine the prevalence of antimicrobial resistance and ESBL production among Escherichia coli and Klebsiella spp from urine samples in a tertiary hospital. This was a cross sectional study conducted at Muhimbili National Hospital in Dar es Salaam, Tanzania.FindingsA total of 270 E.coli and Klebsiella spp urinary pathogens from children and adults isolated from January to March 2010 were included in the study. E. coli and Klebsiella spp isolates were tested for antimicrobial susceptibility by the Clinical and Laboratory Standard Institute's disc diffusion method. These isolates were further screened for ESBL phenotype using cefotaxime and ceftazidime discs. Isolates with reduced sensitivity were confirmed using ESBL E-test strips. Of 270 isolates, 138 (51.1%) were E. coli and 132 (48.9%) were Klebsiella spp. ESBL was detected in 122 (45.2%) of all the isolates. ESBL- producing E. coli strains were significantly more resistance to cotrimoxazole (90.7%), ciprofloxacin (46.3%) and nalidixic acid (61.6%) than strains that did not produce ESBL (p < 0.05). Similarly, ESBL- producing Klebsiella spp strains were significantly more resistance to cotrimoxazole (92.6%), ciprofloxacin (25.0%), nalidixic acid (66.2%), and gentamicin (38.2%) than strains that did not produce ESBL (P < 0.05). Multi-drug resistance was found to be significantly (P < 0.05) more in ESBL producing isolates (90.5%) than non ESBL producers (68.9%). The occurrence of ESBL was significantly higher among isolates from inpatients than outpatients [95 (50.5%) vs. 27(32.9%)] (p = 0.008). The occurrence of ESBL was significantly higher among isolates from children than in adults [84 (54.9%) vs. 38(32.5%)] (p < 0.001).ConclusionsHigh prevalence of ESBL-producing E. coli and Klebsiella spp strains was found among inpatients and children. Most of the ESBL- producing isolates were multi-drug resistant making available therapeutic choices limited. We recommend continued antibiotic surveillance as well comprehensive multi-center studies to address the emerging problem of ESBL-associated infections in order to preserve the continued usefulness of most antimicrobial drugs. Further more conducting molecular studies will help to evaluate the various ESBL types.