Occurrence of K1 and K2 serotypes and genotypic characteristics of extended spectrumβ-lactamases-producing Klebsiella pneumoniae isolated from selected hospitals in Malaysia

Objective:To determine the distribution,phenotypic and genetic background of extended spectrumβ-lactamases(ESBL)-producing Klebsiella(K.)pneumoniae clinical isolates associated with K1 and K2 serotypes in two selected hospitals in Malaysia.Methods:A total of 192 K.pneumoniae isolates were collected and subjected to antibiotic susceptibility,hypermucoviscosity test and multiplex PCR to detect the presence of K1-and K2-serotype associated genes.Multilocus sequence typing(MLST)was performed on ESBL-producing K.pneumoniae isolates presented with K1 and K2 serotypes,followed by phylogenetic analysis.Results:A total of 87 out of 192(45.3%)of the K.pneumoniae isolates collected were ESBL producers.However,only 8.3%(16/192)and 10.9%(21/192)of the total isolates were detected to carry K1-and K2-serotype associated genes,respectively.Statistical analysis showed that K1 and K2 capsular serotypes were not significantly associated with ESBL phenotype(P=0.196).However,they were significantly associated with hypervirulent,as demonstrated by the positive string test(P<0.001).MLST analysis revealed that ST23 as the predominant sequence type(ST)in the K1 serotype,while the ST in the K2 serotype is more diverse.Conclusions:Although the occurrence of ESBL-producing isolates among the hypervirulent strains was low,their coexistence warrants the need for continuous surveillance.MLST showed that these isolates were genetically heterogeneous.

Intestinal carriage of methicillin resistant Staphylococcus aureus and extended-spectrum β-Lactamase-producing Enterobacteriacae in hospitalized and nonhospitalized patients and their clinical implications

Objective:To determine the clinical implication of and intestinal carriage with methicillin resistant Staphylococcus aureus(MRSA) and extended spectrumβ-lactamase(ESBL)-producing Enterobacteriacae.Methods: A total of 180 stool specimens were screened for MRSA and ESBL-producing enterobacteria.Identification of ESBL- producing Enterobacteriacae was done by MicroScan Walk Away 96 system(Dade Behring Inc.,West Sacramento,CA 95691,USA ) and confirmation by double-disc synergy test.MRSA was identified by disc diffusion using 30μg cefoxitin disc and the MicroScan.Results:The rate of fecal MRSA carriage was 7.8% (14/180),35.7%(5 /14) were recovered from surgical wards.Three patients(21,4%) had MRSA recovered from other body sites,and 2(14.2%) had in addition ESBL -producing Escherichia coli(E.coli) and Klebsiella pneumoniae(K.pneumoniae) respectively.Four(28.5%) patients with MRSA fical carriage died. MRSA fecal carriage was recovered from both inpatients and outpatients.Four(2.2%) cases carried ESBL-producing Enterobacteriacae in feces.Three(75%) were from intensive care unit(ICU).One patient had both ESBL-producing E.coli and K.pneumoniae from stool as well as E.coli from tracheal aspirate.Two ICU patients with fecal ESBL died.Conclusion:Fecal screening for MRSA and ESBL of all patients at high risk admitted to different hospital wards and ICUs and implementing infection control measures were recommended.

Prevalence and Potential Risk Factors of Hospital Acquired Extended-Spectrum Beta-Lactamase—Producing <i>Proteus</i>Species

Background: Multidrug resistance and production of extended spectrum β-lactamases (ESBLs) by a large group of bacterial agents in hospitals are to be a matter of scientific concern. Objective: This cross-sectional study was aimed to investigate the prevalence of ESBL producing Proteus species and risk factors associated with hospital acquired infection in addition to study the antibiotics susceptibility patterns of all bacterial isolates from inpatients of four Yemeni general hospitals. Methods: A total of 740 consecutive non-repeat culture isolates were obtained from admitted patients of Al-Kuwait University Hospital, Al-Thowra General Hospital, Al-Jumhori Teaching Hospital, and Military General Hospitals Sana’a city. We used Kirby-Bauer disk diffusion method to detect antimicrobial susceptibility and establish the presence of ESBLs-producing bacteria according to the Clinical and Laboratory Standards Institute guidelines. Results: Out of 740 isolate, 233 (31.5%) were Escherichia coli followed by Staphylococcus aureus 188 (25.4%), Pseudomonas aeruginosa 149 (20.1%), Klebsiella sp. 107 (14.5%), Enterococcus faecalis 25 (3.4%) and Proteus spp. 38 (5.1%). The highest frequencies of ESBLs producing among Proteus sp. were Proteus mirabilis 26 out 38 (68.4%) and Proteus vulgaris 12 out 38 (31.6%). The most effective of antimicrobial susceptibility pattern among Proteus spp. were Imipenem (100%) followed by Pipracillin-Tazobactam (92.3%) for P. mirabilis and (83.3%) for P. vulgaris, while the Amikacin (80.8%) for P. mirabilis and P. vulgaris with (91.7%). Amoxicillin and Cefotaxime were the highest for both species (100%). Conclusion: The prevalence of ESBL-producing Proteus spp. detected in this study is of great concern for public health authorities and a strict adherence of infection control policies and procedures with continuous antibiotics resistance surveillance including antimicrobial management and routine detection of ESBL-producing isolates are very important to prevent nosocomial infections.

Prevalence and characteristics of extended spectrum β-lactamase-producing Escherichia coli from bovine mastitis cases in China

The aim of the study was to investigate the prevalence and characterization of extended-spectrum β-lactamase （ESBL）- producing Escherichia coli isolated from bovine mastitis cases in China. ChromID ESBL agar was used to confirm ESBL-producing E. coli. PCR and DNA sequencing were employed to characterize the genotype of ESBL-producers. Antimicrobial susceptibility was measured by disc diffusion. Overall, 73 of 318 E. coli isolates （22.96％） were identified as ESBL-producers. Of these ESBL-producing E. coli, the prevalence of blaCTX-M and blaTEM-1 was 97.26 and 71.23％, respectively. The predominant CTX-M-type ESBL was CTX-M-15 （65.75％）, followed by CTX-M-14 （10.96％）, CTX-M-55 （9.59％）, CTX-M-64 （5.48％）, CTX-M-65 （4.11％） and CTX-M-3 （1.37％）. This study is the first report of CTX-M-64 and CTX-M-65 in E. coli isolated from bovine mastitis. Furthermore, 72 ESBL-producing E. coli isolates （98.63％） were found to be multidrug-resistance. This study noted high prevalence and rates of antimicrobial resistance of ESBL-producing E. coli isolates from bovine mastitis cases in China.

Multidrug resistance extended spectrum β-lactamase and AmpC producing Escherichia coli isolated from the environment of Bogor Slaughterhouse,Indonesia

To determine the multidrug resistance extended spectrum β-lactamase and AmpC (ESBL/AmpC producing) Escherichia coli (E. coli) isolated from the environment of Bogor slaughterhouse, Indonesia.MethodsA total of 35 samples from 7 locations in slaughterhouse i.e., source of water, slaughtering floor, swab of carcass area floor, swab of evisceration area floor, untreated waste water, treated waste water, drinking water for cattle were collected from March to April 2016. Presence of ESBL/AmpC producing E. coli and susceptibility testing against 8 antimicrobial agents (penicillin G, streptomycin, gentamycin, ciprofloxacin, enrofloxacin, tetracycline, trimethoprim-sulfamethoxazole, and polymyxin B) were detected by disk diffusion test according to Clinical and Laboratory Standards Institute.ResultsESBL/AmpC producing E. coli were identified in 14.3% (5/35) of the collected samples from the environment of Bogor slaughterhouse. ESBL/AmpC-producing E. coli isolates were detected in untreated waste water (n = 3), slaughtering floor (n = 1), and carcass area floor (n = 1). Most of ESBL/AmpC-producing E. coli isolates (80%) showed multidrug resistance phenotypes against at least three classes of antibiotics. The highest incidence of antibiotics resistance was against penicillin G (100.0%) and streptomycin (100.0%), followed by gentamicin (60.0%), trimethoprim-sulfamethoxazole (60.0%), tetracycline (40.0%), ciprofloxacin (40.0%), enrofloxacin (20.0%), and polymyxin B (0.0%).ConclusionsThe transmission of antimicrobial resistant bacteria into the environment may be a potential risk for human health.

Antimicrobial resistance, genotypic characterization and pulsed-field gel electrophoresis typing of extended spectrum β-lactamases-producing clinical Escherichia coli strains in Macao, China

Background The rise of the production of CTX-M class extended spectrum β-lactamases （ESBLs） has been well documented in traveling countries but no data are found for Macao, an international travel city. The objectives of this study were to identify the antimicrobial resistance pattern, and determine the prevalence, genotype and clonal relationship of ESBLs in 209 clinical Escherichia coli strains from Macao, China. Methods Antimicrobial susceptibility test was performed to determine the resistance patterns of the isolates using the disk diffusion method with 17 antimicrobial agents. Phenotypic detection was screened and confirmed according to the Clinical and Laboratory Standards Institute. Genotypic characterization was detected by isoelectric focusing analysis, polymerase chain reaction and sequencing. The clonal relationship between the different ESBL isolates was studied by pulsed-field gel electrophoresis （PFGE）. Results Imipenem and meropenem exhibited 100% susceptible among 209 strains. Overall, 82.3%, 67.3%, 52.9%, 51.2% and 51.0% of the isolates displayed resistance to ampicillin, tetracylcline, ciprofloxacin, sulfamethoxazole trimethoprin and gentamycin. The prevalence rate of ESBLs was 30.1%. Antibiotic resistances were found to be significantly higher among the ESBL producing group compared to non-ESBL producing group. We detected CTX-M-14 to be the major genotypic characterization of ESBLs （76.2%）. Two strains showed indistinguishable patterns by PFGE. Conclusions The prevalence of antimicrobial resistance is alarming high in Macao. Antimicrobial resistance is significantly higher among the ESBL producing group. This study documented CTX-M-14 as the predominant ESBL type. Although indistinguishable pattern was found between two strains, it was too small to decide whether any of the investigated strains was epidemic. Our findings may be also pertinent for other geographic areas undergoing similar travel characteristics to understand the corresponding effects on bacterial populations.

Phenotypic and genotypic detection of extended-spectrum β-lactamase (ESBL) producing Escherichia coli isolated from urinary tract infections in Zabol,Iran

Objective:To investigate the role of a rapid polymerase chain reaction(PCR)assay in comparison with traditional empiric therapy in detection of extended spectrum β-lactamase(ESBL)producer Escherichia coli(E.coli).Methods:Ninety isolates of E.coli from urinary tract infection were collected and screening of ESBL resistance using disc diffusion method,minimum inhibitory concentration(MIC)for ceftazidime and detection of TEM resistant gene by PCR were done.Results:The results of disc diffusion method showed that forty out of ninety E.coli isolates were ESBLs producing organisms.Antibiotic susceptibility of E.coli isolates to 9 antibacterial agents were evaluated.However,all isolated E.coli were resistant to all 9 antibacterial agents by these percentage:ceftriaxon(100%),ceftazidime(100%),amoxicillin(100%),erythromycin(100%),azithromycin(95%),cefixime(87.5%),tetracyclin(87.5%),nalidixic acid(85%)and difloxcain(75%).The abundance of antibiotic-resistant TEM gene according to PCR was 30%.Totally 82.5%of strains tested by MIC were observed as ceftazidime-resistant.Conclusions:We conclude that the TEM gene PCR assay is a rapid,sensitive and clinically useful test,particularly for the early detection of ESBLs-producing E.coli.

Drug resistance of infectious pathogens after liver transplantation

BACKGROUND: Infection in liver recipients is related to high risk of transplantation failure and mortality. Infectious agents isolated from 55 liver recipients from January 2003 through June 2005 were studied to improve the anti-infectious therapy. METHODS: Pathogens were isolated from routine culture. K-B method was used to examine the drug susceptibility. Extended spectrum β-lactamase, AmpC β-lactamase and Van gene in E. coli were examined by the agar-dilution susceptibility test and Nitrocefin test. RESULTS: Thirty-nine of the 55 recipients got infection. The 513 strains of pathogens isolated from 1861 specimens were predominantly Gram negative bacteria and over 40% of them showed resistance to more than 4 drugs. The positive rates of extended spectrum β-lactamse and AmpC β-lactamse production in E. cloacae were 32.4% and 36.8%, in E. coli were 33.8% and 10.5%, but the rates of these 2 bacteria producing both lactamses were 24.3% and 7.0%. The β-lactamse production rates of Enterococcus faecalis and En-terococcus faecium were 8.8% and 11.1%, and the resistance rates to vancomycin were 11.2% and 18.5%, respectively. CONCLUSIONS: Infectious pathogens isolated from liver recipients are potent and multiple drug resistant. ESBLs and AmpC β-lactamases are the major factors associated with Gram negative drug resistance. The infection of En-terococcal species presents as a particular challenge.

Determination of the prevalence of extended spectrum β-lactamase in clinical samples collected from Dehradun City Hospital

Objective:To detect extended spectrumβ-lactamase(ESBL)and determine its prevalence in various clinical samples collected from Dehradun City Hospital.Methods:The samples were first cultured in MacConkey’s agar plates by streak plate method,then identified by Gram staining and biochemical tests.The isolated bacterial strains were then tested for antibiotic susceptibility by Kirby-Bauer method.The ESBL detection is then carried out by double disc diffusion method.Results:Off the 56 samples cultured,21 strains were identified which were six Escherichia coli(E.coli),six Klebsiella,four Proteus,four Pseudomonas aeruginosa(P.aeruginosa)and only one Acinetobacter.Eight out of 21(38.1%)strains including three of E.coli,three of Klebsiella and two of P.aeruginosa,were found to be resistance to all five antibiotics(piperacillin,amikacin,ampicillin,gentamicin,and ciprofloxacin).Initial screening using four antibiotics(cefotaxime,ceftazidime,aztreonam and ceftriaxone)and the final confirmatory test using ceftazidime/clavulanic acid and ceftazidime alone showed that 19.05%of all strains isolated were ESBL producers.Individually,16.67%E.coli,16.67%Klebsiella pneumoniae,25%P.aeruginosa and 100%Acinetobacter were found to be ESBL producers.Conclusions:Antibiotic resistance by ESBL has become a major risk factor worldwide,therefore routine checkup and accordingly prescription are suggested.

Status of ESBL Producing Bacteria Isolated from Skin Wound at a Tertiary Care Hospital in Bangladesh

Background: ESBL producing bacteria are increasing with an alarming rate with a wide range of infections. Objective: The purpose of the present study was to see the status of ESBL producing bacteria isolated from skin wounds. Methodology: This cross sectional study was conducted in the Department of Microbiology at Mymensingh Medical College, Bangladesh from January 2011 to June 2011 for a period of 6 months. All the patients, at any age with both sexes presented with skin wound infection, were taken as study population. Wound swab was taken from all patients. Specimens were processed and bacteria were isolated and identified according to standard procedure. The ESBL status was confirmed by double disc diffusion test (DDDT) and minimum inhibitory concentration (MIC) by agar dilution method by standard procedure according to Clinical Laboratory Standard Institute (CLSI). Antimicrobial resistance was done by disc diffusion method. Result: A total number of 84 wound swabs were taken of which the most common ESBL producing bacteria were Esch. coli (61.5%),?Proteus species (78.3%) and Klebsiella species (88.9%). All the isolates were sensitive to imipenem and nitrofurantoin followed by amikacin (92.9%). Conclusion: In conclusion, ESBL producing E. coli is the most common bacteria causing skin wound infection followed by Proteus species with a reduced sensitivity towards antibiotics.

Antimicrobial Profiles of Selected Gram-Negative Bacteria Recoverable from Sewage and Sludge from Juja and Kibera Informal Settlements of the Larger Nairobi Metropolis

Africa has experienced rapid urban migration in the past two decades. New informal settlements continue to emerge and expand but the sanitation provision of facilities has not improved at the same pace and this poses a serious health concern to the public especially the urban poor. Open sewage systems and sludge-clogged drainage systems as well as soil contaminated with industrial and domestic wastes are possible sources of germs that probably cause clinical infections and epidemics. In this cross-sectional study, we recorded diverse genera of Gram-negative non-fastidious bacteria that included;Escherichia coli (23%), Klebsiella spp (21%), Enterobacter spp (19%), Citrobacter spp (10%), Pseudomonas aeruginosa (8%), Proteus spp (7%), Salmonella (3%), Yersinia spp (3%), Shigella spp (2%), Morganella morganii (2%), Edwardisella spp (1%), Hafnia spp (1%), Serratia marcesence (0.5%), and Acinetobacter baumannii (0.5%). Most of these isolates were resistant to ampicillin while imipenem and ciprofloxacin were the most effective antimicrobial agents. Resistance combination towards ampicillin, trimethoprim, sulfamethoxazole and streptomycin was also noted in recovered isolates (16%). An overall high antimicrobial resistance was recorded among isolates from slum as compared to those recovered from Juja, a middle-class settlement located at the edge of Nairobi metropolis. The prevalence of isolates with a combined resistance to 3rd generation cephalosporins (cefotaxime, ceftriaxone and ceftazidime), gentamicin and ciprofloxacin was the highest among P. aeruginosa isolates (13%) but none of the Yersinia species and Edwardisella tarda exhibited this resistance. Carriage of blaTEM (52%) was most prevalent in all bacteria species followed by blaCTX-M (20%), blaSHV (18%) while blaOXA (17%) was the least common. The phylogeny analysis revealed significant genetic similarity among strains belonging to E. coli, K. pneumoniae, E. agglomerans and P. mirabilis strains but less relatedness was noted among strains belonging to C. freundii. Further analysis showed possible clonal expansion of E. agglomerans and K. pneumoniae within the environmental ecosystems.

Phenotypic and Genotypic Characterisation of Antibiotic Resistance in Escherichia coli, Klebsiella spp., and Listeria monocytogenes Isolates from Raw Meat Sold in Nairobi

Worldwide, the increase in antimicrobial resistance (AMR) is a public health concern. Food-borne associated antibiotic-resistant pathogens can contaminate raw meat during slaughter, transportation, and at sale points. A cross-sectional study was conducted from March 2021 to December 2021 to determine antimicrobial susceptibility patterns and characterize the molecular basis of resistance in E. coli, Klebsiella spp., and L. monocytogenes contaminating raw meat collected from retail outlets in Nairobi. Isolation and identification of the strains were done using the standard culture methods and PCR. Antimicrobial susceptibilities of the recovered strains were determined using disk diffusion while the presence of antibiotic resistance gene determinants;blaTEM, blaCTX-M, blaOXA, sul, and qnrS was done using PCR. Of 270 samples collected, 163 (60%) Escherichia coli, 19 (7%) Klebsiella spp., and L. monocytogenes 3 (1.1%) were recovered. Among Escherichia coli, high antibiotic resistance was found to Erythromycin 161 (98%) and ampicillin 88 (54%) while low resistance was found against imipenem 2 (1%). Similarly, high resistance was found among Klebsiella spp. to Erythromycin 19 (100%) and ampicillin 12 (63%) low resistance to ceftazidime 1 (5%), cefotaxime 1 (5%), aztreonam 1 (5%), and chloramphenicol 1 (5%). One isolate among the three Listeria monocytogenes strains isolated was resistant to Trimethoprim-sulfamethoxazole. No resistance was exhibited to gentamycin by all Klebsiella spp. The prevalence of multidrug-resistant (resistance to three or more classes of antibiotics) isolates was 95/182 (52.2%). The common resistance pattern observed was Erythromycin, ampicillin, tetracycline, and trimethoprim-sulfamethoxazole with a prevalence of 19 (20%). ESBL was confirmed in isolates that harbored: blaTEM (65%), blaCTX-M (44%), blaOXA (33%) while sul and qnrS were detected in 46.7% and 13.6% respectively. Circulation of antibiotic-resistant and MDR isolates found in this study could play a role in the dissemination of AMR among food-borne bacteria and suggest potential food safety and public health risk. Therefore, enhanced surveillance for antibiotic-resistant organisms in raw meat for early detection of emerging resistant bacteria species in the food chain is recommended.