Presumptive Extended-spectrum Beta-lactamase Research Articles

Molecular Characterization and Antimicrobial Resistance Profiling of Extended Spectrum Beta-lactamase (ESBL) Producing Escherichia coli in Bovines from J and K, India

Background: Higher prevalence of ESBL producers is alarming in dairy sector. So limiting antimicrobial use may curtail the selection and persistence of predominant ESBL and conjugative plasmids among strains. The study was aimed to determine the occurrence of extended-spectrum beta-lactamase (ESBL) producing E. coli as well as their genetic diversity, antimicrobial resistance and integrons in bovines. Methods: A total of 180 faecal samples were screened for the presumptive ESBL producing E. coli isolates. All ESBL producing isolates were subjected to the screening test and were confirmed as ESBL producers by double disc. Further, these ESBL producers were tested for the presence of bla genes and those found positive were tested for multidrug resistance (MDR) by disk diffsion. MDR positive isolates were further tested for the presence of intI 1, intI 2 and intI 3 genes. Result: A total of 360 presumptive ESBL producing E. coli isolates were obtained. Out of which, 154 (42.77%) isolates were found to be resistant and were confirmed as ESBL producers. Of 154 ESBL isolates, 120 (77.92%) isolates carried the gene/s screened for blaTEM, blaCTX-M and blaSHV genes are were declared as multi drug resistant. Out of 120 MDR isolates, 59 tested positive for the integron gene.

Relationship Between Multidrug-Resistant Enterobacterales and Obesity in Older Adults.

PurposeThe objective of our study was to determine if obesity is associated with the presence of multidrug-resistant (MDR) bacteria among Enterobacterales.Patients and MethodsThis two-center cohort study included adult hospitalized patients with at least one specimen sampled from any site for bacterial culture yielding an Enterobacterales bacterial species from November 2016 to May 2017. Study groups were stratified by obesity status based on body mass index <30 kg/m2 (non-obese) and ≥30 kg/m2 (obese). The primary outcome was the presence of gram-negative MDR bacteria defined as presumptive extended-spectrum beta-lactamase (ESBL)-producing Enterobacterales (ceftriaxone resistance) or carbapenem-resistant Enterobacterales (CRE). A multivariable logistic regression model was fit to estimate the adjusted odds ratio while controlling for potential confounders.ResultsA total of 366 patients, 238 non-obese and 128 obese, were included. The most common gram-negative bacterial species identified was Escherichia coli (64.2%). There was a higher proportion of gram-negative MDR bacteria in obese versus non-obese patients (18.8 versus 11.3%, P=0.057). Obesity was independently associated with gram-negative MDR bacteria after controlling for confounders (adjusted odds ratio, 1.92; 95% CI 1.03–3.60). The association did not significantly vary by diabetes status (interaction term P=0.792).ConclusionAmong older adult hospitalized patients, obesity was independently associated with the presence of a gram-negative MDR bacteria (presumptive ESBL or CRE) in a culture.

Biofilm formation and extended spectrum beta-lactamase production in Klebsiella pneumoniae isolates from respiratory samples in a tertiary care hospital

Nosocomial and community acquired infections are being caused by Klebsiella pneumoniae (K. pneumoniae) worldwide. K. pneumoniae among gram negative organisms is an important bacterium responsible for site specific infection when they grow as biofilm. Survival of K. pneumoniae is facilitated by biofilm formation that makes it easier to multiply and spread to various sites. Extended–spectrum beta-lactamase (ESBL) producing K. pneumoniae isolates from respiratory samples were used to know the occurrence of biofilm formation conducted in a tertiary care medical college hospital in Mangalore, Dakshina Kannada District, Karnataka, India. To know the occurrence of biofilm formation among extended–spectrum beta-lactamase (ESBL) producing K. pneumoniae isolates from respiratory samples a prospective study has been conducted in a tertiary care medical college hospital in Mangalore, Dakshina Kannada District, Karnataka, India. A total of 87 K. pneumoniae isolates from respiratory samples were characterized according to standard microbiological specific procedures. Screening was done for K. pneumonia Biofilm formation in terms of their antibiotic sensitivity pattern by using standard Kirby Bauer disc diffusion method and presumptive ESBL production by double disk synergy test (DDST). Out of 87 clinical isolates from respiratory samples, 45 (51.72%) were found to be biofilm producers. 29 (33.33%) isolates were ESBL producers and all them produced biofilm. ESBL forming K. pneumoniae isolates had a significantly greater capacity to form strong biofilm (72.4%) than non ESBL producing K. pneumoniae isolates (27.58%). Keywords: Klebsiella pneumoniae, Nosocomial infections, ESBL production, Kirby-Bauer, Biofilm formation.

Biological Hazard on Multidrug Resistance (MDR) of Escherichia Coli Collected From Cloacal Swab of Broiler Chicken on Wet Markets Surabaya

Objective: The purpose of this study was to determine the multidrug resistance (MDR) profile of Escherichiacoli from broiler chicken from Wonokromo Market and Tambahrejo Market Surabaya to some antibioticgroups, namely streptomycin, levofloxacin, cefotaxime, trimethoprim and chloramphenicol.Materials and Methods: Seventy samples were taken from Wonokromo market and Tambahrejo market,each of which had thirty-five samples. Sampling was used cloaca swab technique. Then inoculated on EosinMethylene Blue Agar (EMBA) media, purification of bacteria on EMBA media, and tested biochemicallywith Sulfide Indol Motility (SIM) and Triple Sugar Iron Agar (TSIA) media.Antibiotic sensitivity test was used the Kirby-Bauer method. Muller-Hinton Agar (MHA) media incubation.Then the inhibition zone was measured according to the Clinical and Labortary Standard Institute (CLSI)standards. Detect positive multidrug resistance (MDR) bacteria characterized by resistance to ?3 types ofantibiotics.Results: Detection results of Escherichia coli multidrug resistance (MDR) at Wonokromo market was85.7%, and at Tambahrejo market was 51.4%. Presumptive Extended Spectrum Beta Lactamase (ESBL)producing E. coli at Wonokromo market was 14.3% and at Tambahrejo market was 2.9%.Conclusion: The high level of MDR of Escherichia coli from cloacal swabs of broiler chicken was a threatto public health and the environment, and is an important concern to reduce the rate of its spread

A pilot study using environmental screening to determine the prevalence of Mycobacterium avium subspecies paratuberculosis (MAP) and antimicrobial resistance (AMR) in Irish cattle herds

BackgroundDairy and beef cattle can be reservoirs of many pathogens, including Salmonella and Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of Johne’s disease (JD). Farm environments may provide potential entry points for the transmission of infectious agents into the food chain. Antibiotics are used to treat a wide variety of infections on farms, and administration of antimicrobial agents to cattle is considered to be a driving factor for antimicrobial resistance (AMR). Control of JD and AMR are priority for animal health initiatives in Ireland. A national JD pilot programme was introduced by Animal Health Ireland in 2014, while the national action plan launched by Department of Health and Department of Agriculture, Food and Marine introduced in 2017 aims to improve the surveillance of AMR. The current investigation was undertaken as a pilot study to determine the proportion of herds positive for MAP, Salmonella species (Salmonella spp), commensal Escherichia coli (E. coli), Extended-spectrum beta-lactamase (ESBL) AmpC β-lactamase and carbapenemase-producing E. coli from 157 environmental faecal samples in Irish farms.ResultsMAP was detected in 10.2% of samples collected; on culture in 4 (4.9%) of the dairy herds and from 1 (1.3%) of the beef/suckler herds, and by PCR in 10 (12.3%) and 6 (7.9%) of these herds respectively. All culture positive herds were also positive by PCR. An additional 11 herds were positive by PCR only. Salmonella was not detected, while commensal E. coli were isolated from 70.7% of the samples (111/157) with 101 of these isolates shown to be fully susceptible to all antimicrobials tested. Of the 27 presumptive ESBL AmpC β-lactamase producing E. coli detected, one isolate was resistant to ten antimicrobials, nine isolates were resistant to nine antimicrobials, and four isolates were resistant to eight antimicrobials. Carbapenemase-producing E. coli were not isolated.ConclusionsThe results highlight the importance of monitoring farm environments for Johne’s disease. This disease is a growing concern for dairy and beef producers in Ireland, and sampling the farm environment may offer a useful means to rapidly screen for the presence of MAP. Non-pathogenic common enteric commensal and multiple-drug-resistant E. coli may contribute to AMR acting as a reservoir and transferring resistance to other species/pathogens in the environment.

Antibiogram and hypermucoviscosity pattern among Klebsiella pneumoniae isolates from respiratory samples: A tertiary care hospital study in South India

Klebsiella pneumoniae (K. pneumoniae) is frequently found as normal microbial flora of gastrointestinal tract, nasopharynx and skin, has emerged as a major nosocomial pathogen causing infections of the lower respiratory tract, wounds and urinary tract more commonly opportunistic infections in immune-compromised hosts. Respiratory tract infections are one among the most common nosocomial infections caused by K. pneumoniae. Multi-drug resistant K. pneumoniae (MDR-KP), along with extended-spectrum beta-lactamase (ESBL) producing strains are the cause of concern making it a difficult target for antibiotic chemotherapy. This study was conducted to know the antibiogram pattern and the occurrence of hypermucoviscous in clinical isolates of K. pneumoniae from respiratory samples in a tertiary care medical college hospital in South India. K. pneumoniae isolates were characterized from respiratory tract samples by using standard microbiological procedures showing hypermucoviscosity which were screened for antibiotic sensitivity test by using Kirby Bauer disc diffusion and also by double disk synergy test (DDST) for presumptive ESBL production. In our study we found that, among 87 respiratory samples of K. pneumoniae isolates, 79 (90.8%) were hypermucoviscous and 29 (33.3%) isolates were found to be ESBL producers. This shows that, ESBL producing K. pneumoniae isolates had a greater capacity to produce hypermucoviscosity (100%) than non-ESBL producing K. pneumoniae isolates (86.2%). Keywords: Klebsiella pneumoniae, Antibiogram, Respiratory samples, Hypermucoviscosity, Hypervirulent.

The European Union summary report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food in 2015.

The data on antimicrobial resistance in zoonotic and indicator bacteria in 2015, submitted by 28 EU Member States (MSs), were jointly analysed by EFSA and ECDC. Resistance in zoonotic Salmonella and Campylobacter from humans, animals and food, and resistance in indicator Escherichia coli as well as meticillin‐resistant Staphylococcus aureus in animals and food were addressed. ‘Microbiological’ resistance was assessed using epidemiological cut‐off (ECOFF) values; for some countries, qualitative data on human isolates were interpreted in a way which corresponds closely to the ECOFF‐defined ‘microbiological’ resistance. In Salmonella from humans, high proportions of isolates were resistant to ampicillin, sulfonamides and tetracyclines, whereas resistance to third‐generation cephalosporins was low. In Salmonella and Escherichia coli isolates from fattening pigs and calves under one year of age, resistance to ampicillin, tetracyclines and sulfonamides was frequently detected, whereas resistance to third‐generation cephalosporins was uncommon. For the first time, presumptive extended‐spectrum beta‐lactamase (ESBL)‐/AmpC‐/carbapenemase‐production in Salmonella and Escherichia coli was monitored in humans (Salmonella), meat (pork and beef), fattening pigs and calves. Varying occurrence/prevalence rates of ESBL‐/AmpC‐producers were observed between countries, and carbapenemase‐producing Escherichia coli were detected in single samples of pig meat and from fattening pigs from two MSs. Resistance to colistin was observed at low levels in Salmonella and Escherichia coli from fattening pigs and calves under one year of age and meat thereof. In Campylobacter from humans, high to extremely high proportions of isolates were resistant to ciprofloxacin and tetracyclines, particularly in C. coli. In a few countries, a third to half of C. coli in humans were resistant also to erythromycin, leaving few options for treatment of severe Campylobacter infections. High resistance to ciprofloxacin and tetracyclines was observed in C. coli isolates from fattening pigs, whereas much lower levels were recorded for erythromycin. Co‐resistance to critically important antimicrobials in both human and animal isolates was generally uncommon.

Prevalence of methicillin-resistant &lt;i&gt;Staphylococcus aureus&lt;/i&gt; and extended spectrum β–lactamase producers among bacteria isolated from infected wounds in a tertiary hospital in Ibadan City

Wound colonization by microorganisms is most frequently polymicrobial and incidences of high level resistance among bacterial isolates from wounds have been reported. Methicillin-resistant Staphylococcus aureus (MRSA) and extendedspectrum beta-lactamase (ESBL) producing Gram-negative bacteria both constitute serious challenge to physician in their choice of antibiotic treatment of infections caused by these bacteria. This study determined the antibiotic susceptibility profiles and prevalence of MRSA and ESBL producers among wound bacterial isolates from a tertiary hospital in Ibadan City. Forty (40) clinical bacterial isolates from five wound sources were collected from the Microbiology unit of the University College Hospital (UCH), Ibadan and were authenticated with standard bacteriological techniques. Antibiotic susceptibility test was done by disc-diffusion method using 19 antibiotics belonging to 12 classes. MRSA strains were detected by their resistance to cefoxitin and/or oxacillin antibiotics. Presumptive ESBL production was by double-disc synergy test using 30 μg cefotaxime and ceftazidime around 20/10 μg amoxicillin-clavulanic acid discs. ESBL confirmation was by minimum inhibitory concentration (MIC) using agar-dilution method. The authenticated isolates include Proteus spp (47.5%), Staphylococcus aureus (27.5%), Pseudomonas aeruginosa (12.5%), Klebsiella spp (7.5%), Acinetobacter baumanii (2.5%) and E. coli (2.5%). Distribution of the isolates collected according to wound sources includes: acute soft tissue wounds (35%), leg ulcer (32.5%), surgical wounds (17.5%), burn wounds (12.5%) and diabetic foot ulcer (2.5%). Distributions according to patients’ gender are: male (65%), female (35%), and according to age-groups are: 0 – 19 years (22.5%), 20 – 39 years (35%), 40 – 59 years (32.5%) and ≥ 60 years (10%). All (100%) the isolates were multidrug resistant (MDR) being resistant to ≥ 3 classes of antibiotics. Percentages of isolates resistance to each of the antibiotic include: piperacillin, piperacillin tozobactam and amoxicillin-clavulanic acid were 100%, ceftazidime, cefuroxime, cefixime, aztreonam, sulphamethoxazole-trimethoprim, erythromycin, chloramphenicol and doxycyclin were > 70%, cefoxitin (62.5%), Nitrofurantion (52.5%), ciprofloxacin (45%), ofloxacin (35%), perfloxacin (37.5%), gentamicin (32.5%) and imipenem (2.5%). Of the 11 Staphylococcus aureus collected, 54.5% were detected to be MRSA strains while ESBL production was detected in 55.2% of the Gram negative isolates. This study revealed 100% MDR phenotype constituting high level of MRSA strains (54.5%) and ESBL producers (55.2%) among Gram-positive and Gram-negative bacterial wound isolates respectively. Hence, this calls for caution in the use of extended spectrum antibiotics in treating patients with infected wounds.

Low antibiotic resistance rates in Staphylococcus aureus, Escherichia coli and Klebsiella spp but not in Enterobacter spp and Pseudomonas aeruginosa: a prospective observational study in 14 Swedish ICUs over a 5‐year period

Intensive care units (ICUs) are hot zones for emergence and spread of antibiotic resistance because of frequent invasive procedures, antibiotic usage and transmission of bacteria. We report prospective data on antibiotic use and bacterial resistance from 14 academic and non-academic ICUs, participating in the ICU-STRAMA programme 1999-2003. The quantity of antibiotics delivered to each ICU was calculated as defined daily doses per 1,000 occupied bed days (DDD(1,000)). Specimens for culture were taken on clinical indications and only initial isolates were considered. Species-related breakpoints according to the Swedish Reference Group for Antibiotics were used. Antibiotic resistance was defined as the sum of intermediate and resistant strains. Mean antibiotic use increased from 1,245 DDD(1,000) in 1999 to 1,510 DDD(1,000) in 2003 (P = 0.11 for trend). Of Staphylococcus aureus, 0-1.8% were methicillin resistant (MRSA). A presumptive extended spectrum beta-lactamase (ESBL) phenotype was found in <2.4% of Escherichia coli, based on cefotaxime susceptibility, except a peak in 2002 (4.6%). Cefotaxime resistance was found in 2.6-4.9% of Klebsiella spp. Rates of resistance among Enterobacter spp. to cefotaxime (20-33%) and among Pseudomonas aeruginosa to imipenem (22-33%) and ciprofloxacin (5-21%) showed no time trend. MRSA and cefotaxime-resistant E. coli and Klebsiella spp strains were few despite high total antibiotic consumption. This may be the result of a slow introduction of resistant strains into the ICUs, and good infection control. The cause of imipenem and ciprofloxacin resistance in P. aeruginosa could reflect the increased consumption of these agents plus spread of resistant clones.

Multicentre study of the in vitro activity of cefepime, a broad-spectrum cephalosporin, compared to other broad-spectrum agents

The in vitro activity of cefepime was compared to that of a range of other broad-spectrum agents, using a gradient diffusion MIC method, against 995 recent clinical isolates of Enterobacteriaceae, Pseudomonas aeruginosa, other nonfermentative gram-negative bacilli, staphylococci (except oxacillin-resistant Staphylococcus aureus), streptococci, enterococci, and aerobic gram-positive bacilli. Cefepime had excellent activity against Enterobacteriaceae, including eight presumptive extended-spectrum beta-lactamase producers and 33 stably derepressed mutants of natural cephalosporinases. Activity against Pseudomonas aeruginosa was similar to ceftazidime and superior to cefpirome. Its activity against gram-positive cocci was also good, being more active against staphylococci and only slightly less active against streptococci than ceftriaxone. Cefepime maintained activity against bacteria resistant to aminoglycosides and ciprofloxacin. Enterococci, Bacillus species, Burkholderia cepacia and Stenotrophomonas maltophilia were predicably resistant. Cefepime has a spectrum of activity almost as broad as that of the carbapenems.