Lactamase Production Research Articles

Biofilm formation by carbapenem-resistant Enterobacteriaceae strains isolated from surveillance cultures in Intensive Care Unit patients: A significant problem

Background: Carbapenem-resistant Enterobacteriaceae (CRE) is a serious global public health threat. Antibiotic overuse, improper sanitation and unhygienic practices lead to large scale carriage and rapid spread. Biofilm formation by these strains can lead to far reaching consequences. Material and Method: This is a prospective hospital based study planned for a period of 3 months including all patients admitted to 14 bedded Trauma ICU of a tertiary care hospital in Rajasthan. Rectal swabs were collected from admitted patients and carriage of carbapenem resistant enterobacteriaceae looked for as per CDC guidelines. Screening of the Enterobacteriaceae colonies for carbapenemase production was done by Modified Hodge test. Carbapenem-resistant isolates were also tested for Metallobetalactamase production by phenotypic disc confirmatory test. Biofilm formation testing was done by Tissue culture plate method. Results: A total of 73 patients were screened and 27 CRE isolates were obtained, carriage rate being 37%. A high level of resistance was seen to aminoglycosides, fluoroquinolones and cephalosporins. 100% sensitivity was however seen to Colistin, Tigecycline and Fosfomycin. 5 out of 27 strains showed a positive MHT test. Metallo beta lactamase (MBL) production was seen in 21/25 strains as tested by meropenem and meropenem-edta discs. Of the 27 CRE isolates tested for biofilm production, 13 (48.15%) were found positive. Conclusion: The current study finds out the prevalence of CRE carriage and the biofilm forming potential of these strains among critically ill patients and stresses upon urgent need for stringent infection control measures.

SHV and CTX-M Extended Spectrum Beta Lactamases (ESBL) Producing Bacteria Isolated from Street Foods in and around Chennai, India

Street food vending in India has flourished into a prominent trade especially in the urban and semi urban regions of the country. It is also a major reservoir for the spread of food borne infections caused by microbial pathogens including multidrug resistant organisms. Management of infections caused by multidrug resistant bacteria is difficult. Extended spectrum beta lactamase (ESBL) production contribute to antimicrobial resistance that may result in therapeutic failure. The objective of this study was to analyze for the prevalence of ESBL producing food borne bacteria isolated from street foods sold in and around Chennai. A total of 78 street foods including vegetable samosa (18), chicken samosa (10), panipuri masala (11), panipuri water (31), bhelpuri (8) were tested for the presence of ESBL producing bacteria. The bacteria that were identified included Escherichia coli (55.1%), Staphylococcus sp. (25.6%), Klebsiella sp. (17.9%), Pseudomonas sp. (15.3%) and Enterobacter sp. (11.5%). Multidrug resistance was exhibited by the isolates and ESBL production was found in 16 out of 78 (20.5%) isolates with 9 E. coli, 2 Klebsiella sp., and 5 Pseudomonas sp. Polymerase Chain Reaction (PCR) revealed CTX-M-1(68.75%) and SHV genotypes (31.25%) among the isolates. The 16SrRNA analysis of F10 isolate, confirmed the presence of Pseudomonas sp. in street foods.

Prevalence and Antimicrobial Susceptibility Pattern of Extended Spectrum Beta Lactamase Producers in Gram-negative Urine Isolates at MBN Clinical Laboratories, Kampala Uganda

Introduction: Occurrence of Extended Spectrum beta-lactamase (ESBLs) producing bacteria have presented impediment in treatment choices for urinary tract infections. ESBLs embody a major cluster of lactamases accountable for resistance to novel generations of s-lactam drugs worldwide. The study determined prevalence of ESBL organisms in urine isolates and susceptibility patterns to 13 antibacterial agents. Materials and methods: Two hundred samples were cultured on blood agar, MacConkey agar and incubated at 37°C utmost 48 hours. Isolates identified based on standard bacteriological culture and biochemical characteristics. Drug susceptibility centered on Clinical Laboratory Standard Institute recommended and WHO modified Kirby- Bauer disc diffusion methods. Isolates with reduced susceptibility to Ceftazidime were considered to be possible ESBL producers. Phenotypically confirmed ESBL required use of Ceftazidime in combination with Clavulanic acid. A five milimeter increase zone diameter for Ceftazidime in combination with Clavulanic acid versus its zone tested alone was considered as ESBL. Results: Out of 200 samples, 45 (22.5%) had significant growth, majority Escherichia coli 28 (62.2%), Klebsiella pneumonae 11 (24.4%) followed and Citrobacter fruendii 2 (4.4%). Enterobacter species, Morganella morganii, Proteus mirabilis and Seratia marcescens each 1 (2.2%). Prevalence of ESBLs was 56%. Out of 25 (56%) ESBLs, highest prevalence was among Escherichia coli (15/25; 60%) followed by Klebsiella pneumonae (5/25; 24%) while Citrobacter fruendii, Enterobacter species, Morganella morganii and Proteus mirabilis each had (1/25; 4%). Susceptibility was highest to Imipenem 22 (88%) and least to Ampicillin, Amoxicillin-Clavulanic acid and Tetracycline each 1 (4%). No susceptibility to Cefuroxime and Ceftazidime was observed. Conclusion: The study showed that resistant ESBLs generating bacteria were present among urine isolates. We recommend that ESBLs bacteria isolated in urine be treated based on antibiotics susceptibility, continuous surveillance to guide correct treatment for urinary tract infection and to prevent the occurrence of multi drug resistant bacteria. This should be reflected in the policies developed by the Ministries of health to promote rational use of antibiotics.

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

Molecular detection of methicillin resistant Staphylococcus aureus harbouring β- lactamase resistance genes isolated from different sources of infections

Background and objective: The detection and investigation of methicillin resistance staphylococci specifically S. aureus in clinical microbiology setting is very helpful both for informing the appropriate treatment of individual patients and also for the surveillance of these organisms. This study aimed at the rapid molecular detection of methicillin resistant staphylococci harbouring β-lactamase gene and determination of the efficiency of m-PCR through comparison with uniplex PCR. Methods: Standard microbiological techniques were applied for the determination of the presence of methicillin resistant S. aureus in samples recovered from different body sites of patients who attended Al-Kadhumyhia Teaching and Baghdad Teaching Hospitals. The resulting methicillin resistant S. aureus (MRSA) isolates were subjected to uni and multiplex PCR amplifications for detecting the existence of mec A gene and β-lactamase (TEM) resistance gene. Results: Half of the cases involved were found to be caused by MRSA. All the tested isolates showed positive amplification bands for the presence of mec A gene and only 48.8% of these harbored TEM gene. The obtained results revealed high sensitivity of universal bacterial and TEM primers expressed as 97.6% and 100% respectively, while the sensitivity of mec A primer was limited to 60%. Conclusion: The phenotypic identification of MRSA revealed a higher incidence rate and that different molecular techniques can yield conflicting results and it can also be concluded that resistant due to beta- lactamase production can be a crucial factor added to the previously settled methicillin resistant due to mec A gene.

Detection of prevalence of metallo-beta lactamases in clinical isolates of imipenem resistant Pseudomonas aeruginosa from neonatal septicaemia cases in a tertiary hospital in Odisha, India

Background: Pseudomonas aeruginosa is a clinically troublesome gram-negative pathogen that causes both opportunistic infections and nosocomial outbreaks. Metallo beta lactamase have recently emerged as a worrisome resistance mechanism. Carbapenems had been the drug of choice for the infections caused by most penicillin- or cephalosporin-resistant gram-negative bacteria due to its broad-spectrum activity and stability to hydrolysis by most beta-lactamases. This does not hold good anymore due to rapid uprise of MBL producing strains. The current research covered 163 hospitalized cases of neonatal septicaemia from which Pseudomonas aeruginosa is isolated in the Paediatric Department of KIMS, Bhubaneswar. The study aimed at detecting the prevalence of metallo-beta lactamases in clinical isolates of imipenem resistant Pseudomonas aeruginosa from neonatal septicemia cases and to establish the antibiogram of Imipenem-resistant P. aeruginosa these cases. Methods: Clinical samples obtained from suspected cases of neonatal septicemia were first cultured by conventional methods and then identification was done by VITEK-2 instrument. Metallo beta lactamase (MBL) production was done by combined disc synergy test (CDST) using imipenem and EDTA (CDST-IPM) and double disc synergy test (DDST) using IPM and EDTA (DDST-IPM). Results: Among 1510 processed clinical specimens from cases of neonatal septicaemia; 637 (42.18%) showed positive growth of various clinically significant pathogens. Out of them in 163 (25.58%) cases Pseudomonas spp. was isolated. Of these, a total of 95 (58.28%) Pseudomonas spp. was found resistant to imipenem. All imipenem-resistant Pseudomonas isolates were positive for MBL by CDST imipenem-EDTA (CDST-IPM) method, whereas 89 (93.68%) were positive by DDST-IPM method, respectively. Pseudomonas aeruginosa was mostly isolated from endotracheal tube aspirate (57.89%) followed by pus (56.41%). Out of the 95 cases of MBL-producing Pseudomonas; 46 (48.42%) isolates showed the maximum susceptibility to piperacillin-tazobactam combination. All MBL-producing Pseudomonas isolates were resistant to ceftriaxone.Conclusions: MBL-producing Pseudomonas is found to be highly prevalent in our hospital, which is one of the major causes of multidrug resistance and need regular surveillance and strict adherence to a robust antibiotic policy.

Prevalance of Extended Spectrum Beta Lactamases (ESBL) Producing Escherichia Coli Isolated From Clinical Samples at Tertiary Care Hospital Peshawar

Escherichia coli is Gram negative, facultative and non sporulating rod shaped bacteria. It is commonly inhabitant of the gastrointestinal tract of humans and animals. E. coli cause diseases like urinary tract infection cholecystitis, cholangitis and traveler’s diarrhoea and the UTI which is more prevalent worldwide. E. coli cause complication like Hemolytic Uremic Syndrome which leads to renal failure, Thrombotic Thrombocytopenic Purpura, septicemia and peritonitis. Extend spectrum beta lactamase enzyme produce by E. coli which capable of hydrolyzing first and third generation cephalosporin, and is inhibited by beta lactamase inhibitor. A total of 150 clinical samples (blood, urine, wound swab, body fluids) were collected from Post Graduate Lady Reading Hospital Peshawar. Different media used were Nutrient agar, MacConky agar and Cysteine, Lactose and Electrolyte-Deficient agar. E. coligive pink colonies on MacConky agar because it is lactose fermenter. For further confirmation different biochemical tests were performed like triple sugar iron, Indole, and citrate utilization tests. The antibiotics susceptibility and resistivity was checked by disk diffusion method and different antibiotics were used. For ESBL detection combined disk method was performed. In the clinical samples the percentage of Gram positive bacteria in blood was (20%), urine (14.2%), wound swab (83.3%), and body fluids (8%), and the Gram negative in urine was (80%), blood (7%), wound swab (10%), and body fluids (0%). E. coli was more prevalent in urine which was 25(35.71%) and ESBL producing E. coli was 5(20%). The ESBL producing E. coli was resistant to ciprofloxacin (100%), amikacin (40%), amoxicillin+clavulanic acid (40%), levofloxacin (80%), tazobactum+pipracilline (20%), gentamycin (100%), trimethoprim (60%), cefotaxime (100%) and meropenem (0%). Sensitivity toward levofloxacin (20%), tazobactum+pipracilline (80%), gentamycin (0%), trimethoprim (40%), cefotaxime (0%) and meropenem (100%), ciprofloxacin (0%), amikacin (60%), amoxicillin+clavulanic acid (60%). The most effective antibiotic against ESBL producing E. coli was Meropenem while least effective antibiotics against ESBL producing E. coli were Gentamycin and Ciprofloxacin.

Extended spectrum and metalo beta-lactamase producing airborne Pseudomonas aeruginosa and Acinetobacter baumanii in restricted settings of a referral hospital: a neglected condition

BackgroundFrequently encountered multidrug-resistant bacterial isolates of P. aeruginosa and A. baumannii are common and prevalent in a hospital environment. The aim of this study was to determine the prevalence and pattern of antibiotic resistance, extended spectrum and metallo beta-lactamase producing P. aeruginosa and A. baumannii isolates from restricted settings of indoor air hospital environment.MethodsA hospital-based cross-sectional study was conducted in Wolaita Sodo University Teaching and referral Hospital, Ethiopia from December 1/2015 to April 30/2015. The Air samples were collected from delivery room, intensive care unit and operation theatre of the hospital by active, Anderson six slate sampler technique during the first week of the months, twice a week during Monday’s and Friday’s. Standard microbiological procedures were followed to isolate P. aeruginosa and A. baumannii. Susceptibility testing was performed on isolates using the Kirby-Bauer disk diffusion technique. Extended spectrum beta lactamase production was detected by double disc synergy test and Imipenem-resistant isolates were screened for producing Metallo-beta lactamase.ResultsA total number of 216 indoor air samples were collected from the delivery room, intensive care unit, and operation room. Correspondingly, 43 A. baumannii isolates were identified (13 from delivery room, 21 from intensive care unit and 9 from operation room). Likewise 24 P. aeruginosa isolates were obtained (4 from delivery room, 13 from intensive care unit and 7 from operation room). Extended spectrum beta lactamase and metalo-beta lactamase production were observed in 24 (55.8%) and 13 (30.2%) isolates of A. baumannii respectively, whereas P. aeruginosa showed 15 (62.5%) extended spectrum beta lactamase and 9 (37.5%) metallo-beta lactamase production.ConclusionsExtended spectrum beta lactamase and metallo-beta lactamase producing bacteria in hospital air is a new dimension for specific setting of the study area where antimicrobial resistance is increasing and surgical site infection is prevalent. So, identification of these microorganisms has a great role in reducing the burden of antibiotic resistance and could also provide a significant input for framing hospital infection control policies.

Investigating of four main carbapenem-resistance mechanisms in high-level carbapenem resistant Pseudomonas aeruginosa isolated from burn patients

BackgroundPseudomonas aeruginosa is an opportunistic pathogen involved in many infections. Carbapenem-resistant P.aeruginosa has emerged as an important cause of infection in different hospitals worldwide. We aimed to determine frequencies of the four main resistance mechanisms [metallo-beta lactamase (MBL) production (blaIMP, blaVIM, blaSPM and blaNDM), overproduction of the MexAB–OprM and MexXY efflux pumps, overproduction of chromosome-encoded AmpC β–lactamase, and reduced OprD expression] in high-level carbapenem-resistant P.aeruginosa isolated from patients with burns. MethodsIn a descriptive study, 107 P. aeruginosa isolates were collected from patients with burn injuries and tested for antibiotic susceptibility, by an E-test for carbapenems, an E-test for metallo-β-lactamase producer isolates, and PCR to detect MBL genes. Furthermore, high-level carbapenem-resistant isolates were tested by real-time PCR for the expression levels of the mexB, mexY, ampC, and oprD genes. ResultsAmongst all P. aeruginosa isolates, 78.5%, 46.7%, and 15% were imipenem-, meropenem-, and doripenem-resistant, respectively; 72% of isolates were multidrug-resistant. The blaIMP and blaVIM genes were detected in 17.9% and 1.2% of isolates; respectively. The blaSPM and blaNDM genes were not observed. Among the resistant isolates, mexB overexpression (63.2%) was the most frequent mechanism, followed by mexY overexpression (52.6%), ampC overexpression (36.8%), and reduced oprD expression (21.1%). ConclusionEmerging antimicrobial resistance in burn wound bacterial pathogens is a serious therapeutic challenge for clinicians. In the present study, most of the isolates were MDR. This finding indicated an alarming spread of resistant isolates and suggested that infection control strategies should be considered. Resistance to carbapenems is influenced by several factors, not all of which were evaluated in our study; however, the results showed that production of MBLs and overexpression of the mexB gene were the most frequent mechanisms in carbapenem-resistant isolates.

Multiple Liver Abscesses due to Citrobacter koseri: An Extremely Rare Presentation

Pyogenic liver abscess (PLA) of the liver is relatively rare. Citrobacter is a very rare cause of liver abscess. We report a case of multiple liver abscess due to Citrobacter koseri. This is the 3rd reported case with liver abscess and Citrobacter koseri. A 61 years old woman presented to the ER with abdominal pain, fever, and progressive jaundice of 2 weeks duration. She had history of NHL which presented with biliary obstruction that required biliary stent placement 3 years back. The biliary sent was not removed as the patient was lost to follow up. She was hypotensive requiring IV fluid boluses and pressor support. Laboratory studies: white blood count 25.16 k/μL, serum creatinine 1.3 mg/dl, total bilirubin 2.5 (mg/dL), direct bilirubin 2.0 (mg/dL), alkaline phosphatase 345 (IU/L), ALT 57 (IU/L), and AST 152 (IU/L). U/S of the abdomen showed extensive intrahepatic biliary dilatation and multiple hypoechoic structure in the liver. CT of the abdomen showed lesions scattered throughout the liver suscpecious of abscesses. Blood culture was positive for Citrobacter koseri. Patient underwent liver biopsy with aspiration pus and three core biopsies. Citrobacter koseri was isolated from the pus. Pathology was negative for malignancy. The patient was treated with IV Imipenem and oral ciprofloxacin for 6 weeks and had remarkable clinical improvement. Follow up imaging studies with U/S showed complete resolution of the abscesses. Pyogenic Liver Abscesses due to Citrobacter koseri is a rare infection; we are aware of only two reported cases in the literature. Therefore, little is known about Citrobacter liver abscess in terms of incidence, associated underlying diseases, and outcome. Treatment of citrobacter koseri infection in general could be complicated by rapid development of resistance particularly if monotherapy with a cephalosporin is started. This is mainly due to due to high level of beta lactamase and cephalosporinase production. Thus, combination therapy is recommended for initial empirical treatment of Citrobacter koseri infection.

Nosocomial bacterial infections and their antimicrobial susceptibility patterns among patients in Ugandan intensive care units: a cross sectional study

BackgroundThe intensive care unit (ICU) admits critically ill patients requiring advanced airway, respiratory, cardiac and renal support. Despite the highly-specialized interventions, the mortality and morbidity is still high due to a number of reasons including nosocomial infections, which are the most likely complications in hospitalized patients with the rates being highest among ICU patients.MethodsIn this cross-sectional study of 111 adult patients admitted to 2 of the ICUs in Uganda, we set out to describe the commonest bacterial infections, their antimicrobial susceptibility patterns and factors associated with development of a nosocomial infection.ResultsKlebsiella pneumoniae (30%), Acinetobacter species (22%) and Staphylococcus aureus (14%) were the most frequently isolated bacteria. The prevalence of multidrug resistant bacterial species was 58%; 50% Escherichia coli and 33.3% Klebsiella pneumoniae were extended spectrum beta lactamase or AmpC beta lactamase producers and 9.1% Acinetobacter species were extensive drug resistant. Imipenem was the antibiotic with the highest susceptibility rates across most bacterial species. Institution of ventilator support (P 0.003) and severe traumatic brain injury (P 0.035) were highly associated with the development of nosocomial infections.ConclusionDue to the high prevalence of multi drug resistant (MDR) and extensive drug resistant bacterial species, there is a need for development of strong policies on antibiotic stewardship, antimicrobial surveillance and infection control to help guide empirical antibiotic therapy and prevent the spread of MDR bacteria and antibiotic drug resistance.

Screening for Extended - Spectrum β-Lactamases, AmpC and Carbapenemases Producing Bacteria among Adult and Pediatric Intensive Care Unit Patients in Two University Hospitals in Egypt

Background: Emerging β-lactam resistant gram-negative bacteria (BLR-GNB) is a problem worldwide, particularly in the Intensive Care Units (ICUs). Objective: The aim of the present work was to evaluate the rate of carriage and the acquisition rate of BLR-GNB at one adult and one pediatric ICU in Alexandria University Hospitals, Egypt. Methodology: One hundred adult and 50 pediatric intensive care unit patients were included in the study. Rectal and nasal swabs were inoculated onto chromID ESBL (BioMerieux). The rate of carriage and the acquisition rate of BLR-GNB were calculated in both ICUs. All identified GNB were tested for ESBL, AmpC-β lactamase and carbapenemase production using combined disc, AmpC disc, and modified Hodge test (MHT) respectively. Results: Eighty eight percent of adult and 74% of pediatric patients were BLR-GNB carriers. The acquisition rate of BLR-GNB in adult & pediatric ICUs 48 hrs after admission was 42.9% and 35% respectively. Out of the 150-screened patients, 280 BLR-GNB isolates were identified where E. coli was the most commonly isolated (45%) followed by K. pneumoniae (35%). Conclusions: The high rate of BLR-GNB carriage calls for routine surveillance, which will be helpful for monitoring the spread of drug resistance in ICU settings.

Expanded spectrum β–lactamase producing Escherichia coli isolated from chickens with colibacillosis in Egypt

Throughout the world, expanded spectrum β-lactamases (ESBL) are increasing among clinical isolates of Enterobacteriaceae, both in humans and animals. Unfortunately, there is a paucity of data on ESBL or Ampicillin class C β-lactamase (AmpC) in Egypt, although antimicrobial consumption is high in this developing country. This study aims to characterize the resistance mechanisms to expanded spectrum cephalosporins among resistant veterinary Escherichia coli isolates in Egypt. We investigated 50 clinical multi-resistant E. coli strains isolated from 20 chicken farms for production of ESBL or AmpC. Antibiotic susceptibility was tested by Clinical and Laboratory Standards Institute (CLSI) disk diffusion and ESBL confirmatory tests. PCR and sequencing were performed to screen for plasmid mediated ESBL genes and genes encoding AmpC β-lactamases. All the isolates were phylogentically classified, investigated for harboring class 1 integrons, and genotyped by amplified fragment length polymorphism (AFLP). Three strains showed ESBL and 6 strains AmpC phenotypic patterns, respectively, with confirmed ESBL genes of blaTEM-57, blaSHV-12, blaCTX-M-14, and blaCMY-2 for AmpC producing strains. All ESBL strains belonged to phylogroup D with different clones isolated from different flocks, while most of the AmpC strains belonged to phylogroup B1 (4/6) and were assigned to the same genotype distributed in 2 different farms. Class 1 integrons were disseminated in 60% of all tested strains and in 100% of ESBL and AmpC strains. These results highlight the antimicrobial resistance problem in Egypt, caused in all probability by unwise use of antimicrobials in animal husbandry. The results call for a nationwide surveillance program to monitor antimicrobial resistance.

Incidence microbiological profile and drug resistance pattern of uropathogens causing asymptomatic bacteriuria among below poverty line diabetic male patients

Introduction: With the prevalence of DM increasing among rural population in developing countries, factors associated with diabetes and its complications also becomes more important. More than half of diabetic patients with ASB have upper urinary tract involvement and the frequency of symptomatic UTI had been significantly higher. Symptomatic UTIs tends to be more common in diabetic subjects with ASB than in those without ASB. Although ASB is of major concern in diabetic population, the long-term consequences of ASB in patients with DM are poorly documented, Almost all studies were performed among elderly women with type 2 diabetes and there is very little information on the occurrence of ASB among BPL diabetic males in our local setting. Hence in the present study the incidence and etiology of ASB among BPL diabetic male patients was monitored along with the resistance pattern of bacterial isolates to antimicrobial agents.Materials and methods: Clean catch voided midstream urine samples were collected from 1131 BPL Diabetic male patients enrolled for the study. Wet film of centrifuged urine was performed to detect the presence of pus cells, epithelial cells, erythrocytes, microorganisms, cast. Culture was performed using standard loop method and antimicrobial susceptibility of the isolates was studied using Kirby Bauer disc diffusion method following CLSI guidelines.Results: Out of 1131 BPL diabetic male patients screened for ASB, 155 (13.7%) were culture positive. Among the uropathogens Gram negative bacilli was the most commonest type (72.7%) and the most prevalent organisms isolated was Klebsiella spp (35.2%), Enterococcus spp (22.4%), followed by E. coli (19.4 %) ,Pseudomonas aeruginosa ( 7.3%), etc.,. 87.5% of E.coli isolated were ESBL, followed by 77.6 % of Klebsilla spp and 11.1 % Enteroabacter spp. Pseudomonas aeruginosa reported in this study were 100% ESBL and 16.6% Metallo β lactamase (MBL) producers.8.1 % of Vancomycin resistant Enterococcus (VRE) was also found in this study.Conclusion: This study demonstrated a high occurrence of ASB in BPL diabetic males (13.7%). Klebsiella was the most commonest uropathogen found in our study followed by Enterococcus , E.coli and Pseudomonas. E.coli and Pseudomonas showed high rates of drug resistance. Nitrofurantoin and Amikacin was the most effective drugs for majority of the isolates. Hence routine monitoring and screening for ASB in this population is essential. Moreover patients in rural parts of developing countries with diabetes has to be sensitized about the complications of ASB and regarding maintenance of their glycemic control which is of major importance in prevention of the condition.

Drug resistance pattern analysis of various organisms isolated from neonatal intensive care unit of a tertiary care hospital in Odisha, India

Background: Neonatal septicaemia is characterized by systemic signs and symptoms of generalized bacteraemia with a positive blood culture in the first four weeks of life. This study was designed to detect Multi Drug Resistant pattern from organisms in neonatal septicaemic cases. The aim of the study is to find the antibiotic sensitivity pattern and drug resistance pattern (ESBL, MBL, Carbapenemase, AmpC β-lactamase, MRSA) among the organisms isolated. Methods: This prospective study was carried out in the Department of Microbiology in association with Dept. of Paediatrics and NICU, of Kalinga Institute of Medical Sciences, Bhubaneswar, Odisha, India during the period from November 2012 - April 2014. The study was conducted on 250 patients. The inclusion criteria are neonates with suspected septicaemia, admitted to NICU, of KIMS, Bhubaneswar, Odisha, India. Two ml of venous blood from each neonate was collected & cultured by automated method. Phenotypic confirmatory tests for ESBL production and MBL detection was done as per CLSI guideline. MRSA detection was done by Cefoxitin disk (30 mcg) screen test. Results: All gram positive pathogenic isolates were sensitive to Linezolid, Tigecycline and Vancomycin. Maximum resistance was seen against Benzylpenicilin and Ampicillin. Among the gram-negative isolates maximum antibiotic sensitivity was observed for Tigecycline, Levofloxacin and amikacin & resistance was maximum for cefadroxil and Ampicilin. Among the gram negative bacterial pathogens ESBL production was maximum by E.coli (75%) and Burkholderia cepacia (75%) followed by Enterobacter cloacae (66.7%) and Acinetobacter iwoffi (50%). Metallo- beta- lactamase production was seen maximum in Acinetobacter iwoffi (100%) followed by Burkholderia cepacia (75%). Methicillin resistance was seen maximum by S. epidermidis (58.3%) followed by S. haemolyticus, S. aereus, S. warneri and S. hominis.Conclusions: This study facilitated the screening of MRSA, ESBL and MBL producing pathogens which are an emerging problem. Long term surveillance is needed to combat this emerging global challenge.

Emergence of multi drug resistance strains causing diabetic foot infection in Salem, Tamil Nadu, India

This study was to assess the emergence of mecA and bla CTX-M in multidrug resistant MRSA/MRSE and ESBL strains in diabetic foot infections. A total of 160 pus samples were collected from the diabetic foot care hospitals, Salem, Tamil Nadu, India. The samples were processed for microbiological investigations and further evaluating the resistant strains. The bacterial strain Staphylococcus aureus was the predominant organisms isolated from the sample. Cefoxitin, Oxacillin and Vancomycin were found to be the most effective antimicrobial agents for therapy of gram positive organisms while Meropenem, Piperacillin, Cefoperazone/Sulbactam, Piperacillin/ Tazobactam and Amikacin were found to be the most effective antimicrobial agents for the gram negative organisms. In this study, four isolates of Methicillin resistant Staphylococcus aureus (MRSA) strains, one isolate of Methicillin resistant Staphylococcus epidermidis (MRSE) and five isolates of Pseudomonas aeruginosa were found to be extended spectrum of β - lactamase (ESBL) producers by phenotypic method. These strains subjected to genotypic confirmation confirmed the presence of resistant genes namely bla CTX-M in ESBL and SCC mecA in Staphylococcus spp. The 16S rRNA partial gene sequencing and phylogenetic analysis of the multi drug resistant organisms Pseudomonas aeruginosa (APS01) and Staphylococcus epidermidis (APS02) was confirmed the resistant pattern in these strains. The resistant organism was eradicated only by the effective management of this infection; knowledge on the causative agent causing cruel diabetic foot infections will be very useful in selecting the appropriate therapy.

Evaluation of Stability of Cefamandol and Ceftazidime with Clavulanic Acid Against Extended Spectrum ?- Lactamase

The aim of this study is to evaluate in-vitro activity of Cefamandol (Cfm) and Ceftazidime (Cfz), in combination with Clavulanic acid (CA) against ten complicated multiresistant uropathogenic E.coli .One hundred clinical strains were isolated from patients with chronic urinary tract infections (UTIs), these isolates were identified by the Api identification systems. The antimicrobial susceptibility tests were determined by Kirby-Bauer method, all of them were sensitive to Imipenem (Imp). Ten strains were chosen for the present study, they were resistant to Ampicillin (Amp), Amoxicillin (Amo), Carbenicillin (Cb), Ticarcillin (Tic), Azlocillin (Azl), Amoxicillin\ Potassium Clavulanate {Augmentin(Amc)}, (Amo\CA), Ticarcillin\ Potassium Clavulanate {Timentin} (Tic\CA) ,Cefazolin (Cfo) ,Cefaloridin (Cfr), Cefamandol, (Cfm),Cefoxitin, Ceftazidime (Cfz), Cefixime (Cxm), Cefoperazone( Cfp) and Aztreonam (Atm), also resistant to other antibiotics, Tetracycline(Tc),Cloramphenicol(Cm),Gentamycin(G),Amikacin (Amk), Ciprofloxacin (Cip) and Trimethoprim. 50% of the isolates were resistant to Nalidixic acid and Rifampicin. The minimum inhibitory concentrations of Cefamandol and Ceftazidime were determined, by tube method. Transfer of plasmids were done by direct conjugation test to sensitive standard E.coli ,cell free β- lactamases were prepared and detected by macro-iodometric method. The activity of each cell free ß– lactamases extract against Cfm and Cfz were determined by disks diffusion method (microbiological Masuda method). Excellent activities were obtained against these strains when Cfm and Cfz, combined with CA, therefore complete zones of inhibition were obtained indicated the prevalence of extended spectrum β- lactamases in E.coli. The stability of Cfm and Cfz in the presence of CA were useful in the treatment of chronic urinary tract infections caused by multiresistant β- lactamase (ESBL) producer E.coli.
 Key words: Extended spectrum β- lactamases, Imipenem, Aztreonam, Ceftazidime.

Prevalence of Extend Spectrum Beta Lactamases Producing Enterobacteriaceae and their Antibiotic Susceptibility in Lome, Togo

Prevalence of Extend Spectrum Beta Lactamases Producing Enterobacteriaceae and their Antibiotic Susceptibility in Lome, Togo

CTX-M-15 and OXA-10 beta lactamases in multi drug resistant Pseudomonas aeruginosa: First report from Pakistan.

Pseudomonas aeruginosa is an emerging threat to public health worldwide due to their rapid development of drug resistance including beta-lactamases. The present study was designed to investigate the incidence of β-lactamases and genotypic pattern of CTX and OXA in the clinical isolate of multidrug resistant P.aeruginosa. In this study a total of 102 MDR P.aeruginosa isolates obtained from Lady Reading Hospital, Peshawar, Pakistan were subjected to extended spectrum beta lactamase (ESBL), metallo beta lactamase (MBL) and plasmid mediated β-lactamase (AmpC) detection using phenotypic and molecular methods. Furthermore, sequencing of CTX and OXA gene was performed. Out of 102 MDR P.aeruginosa isolates, 71 (69.6%) were beta lactamase producers. The incidence of ESBL, MBL and AmpC in clinical isolates of P.aeruginosa was found to be 23.94%, 40.84% and 35.21% respectively. Co-production of ESBL and AmpC were also observed in some isolates. There were 14 (19.71%) CTX-M-15 harboring isolates which were ESBL (64.28%), MBL (21.42%) and AmpC (14.28%) producer. Co-production of ESBL/MBL (14.28%), ESBL/AmpC (14.28%) and MBL/AmpC (14.28%) were also observed in the CTX M-15 harboring isolates while 12.28% isolates were not ESBL, MBL or AmpC producer. OXA-10 was detected in 8 (11.26%) isolates which were ESBL (12.5%), MBL (37.5%) and AmpC (12.5%) producer. OXA 10 isolates also exhibit co-production of ESBL/AmpC (12.5%) and MBL/AmpC (12.5%). All CTX-M-15 carried the class A β-lactamase conserved domain while OXA-10 harbored conserved domain of class D β-lactamase. The current study for the first time reported and characterized the CTX-M-15 and OXA-10 among MDR P.aeruginosa isolates from Pakistan. Further efforts are needed to understand the molecular mechanism of drug resistance with CTX and OXA harboring P.aeruginosa isolates.

Prevalence of carbapenem resistant bacterial strains isolated from different clinical samples: study from a tertiary care hospital in Kathmandu, Nepal

Background Carbapenems are considered as drugs of choice for the treatment of the infections caused by drug resistant bacteria. However, in the recent years the prevalence of carbapenem resistant gram negative bacteria has increased significantly. The main objective of this study was to determine the prevalence of carbapenemase producing gram negative bacteria among all the clinical isolates.Material and methods A total of 3246 non-repeated, different clinical specimens from patients attending Kathmandu Model Hospital, from July 2013 to January 2014 were cultured and the gram negative bacterial isolates obtained were subjected to identification with the help of colony morphology, Gram’s stain and conventional biochemical tests. Kirby-Bauer disk diffusion technique was used to perform antimicrobial susceptibility testing. Phenotypic confirmation of carbapenemase and AmpC beta-lactamase production was done by combined disc method.Results 890 samples showed the growth of bacterial pathogens. Out of total 769 gram negative bacteria, 57 were found to be carbapenem resistant. Of which, highest number (47) of the isolates were found to be metallo-β lactamase (MBL) producers. Six bacterial isolates produced both (Klebsiella pneumoniae carbapenemase) KPC and MBL, whereas only one isolate was found to be positive for both MBL and AmpC. Three bacterial strains showed carbapenem resistance due to over production of AmpC β-lactamase.Conclusion Among carbapenem resistant gram negative bacteria, MBL was present as the major enzyme responsible for resisting carbapenem antibiotics.