Clinical Laboratory Standards Institute Research Articles

Characteristics and Antibiogram Studies of Bacteria Associated with Vegetables Stored In Raffia Baskets in Nigeria

Raffia baskets are used in rural communities to preserve fresh vegetables for a given period of time. This method of storage is aimed at extending the shelf life of the vegetables until they are used. Thus, this study was aimed at determining the microbial succession during storage and susceptibility pattern of bacterial isolates from stored leafy vegetables to various antibiotics. Five (5) leafy vegetables namely, Bitter leaf (Vernomia anydalira), Water leaf (Talinum triangulare), Fluted pumpkin leaf, (Telfairia occidentalis), Okazi leaf (Gnetum africana) and Scent leaf (Ocimum gratissimum) were obtained from the Nigerian Stored Products Research Institute (NSPRI) farm, Port Harcourt. These vegetables were stored in a raffia basket for fourteen (14) days while another set of the vegetables were left in the open air as control. This set up was monitored for changes to occur. Standard microbiological techniques were employed for the various analyses of the vegetable samples. Both cultural and molecular characterizations of the isolates were done. The disc diffusion method was used in determining the susceptibility pattern of the bacterial isolates. The results of the predominant microorganisms identified were of the genus; Bacillus, Pseudomonas, Enterococcus, Enterobacter, Bordetella, Staphylococcus, Myroides, Escherichia, Serratia, Micrococcus and Acetobacter. Also, Bacillus species occurred in all the vegetables while Enterococcus faecalis, Acetobacter orientalis, Bordetella pertussis, Myroides xuanwuensis and Bacillus flexus were isolated on the third day of storage from the vegetables. The total heterotrophic bacterial counts ranged from 1.8 x106 cfu/g to 1.25 x107 cfu/g (bitter leaf), 9x105 cfu/g to 9.0 x106 cfu/g (Scent leaf), 7 x105 cfu/g to 1.88 x107 cfu/g (Okazi), 3.2x106 cfu/g to 1.05x107 cfu/g (Fluted pumpkin leaf) and 8.4 x106 cfu/g to 2.04 x107 cfu/g (water leaf). Antibiogram of bacterial isolates revealed that they were resistant to Augmentin and Ceftazidime according to Clinical Laboratory Standard Institute schemes. Combination of the antibiotics showed that most of the isolates were 100% susceptible to ciprofloxacin + erythromycin, and ciprofloxacin + ceftriaxone. The study revealed that for all the vegetables, those stored in the basket lasted longer and were fresher compared to those kept in the open space. This study revealed that the raffia baskets are suitable for storage and preservation of leafy vegetables, but microbial succession occurred as a result of increase in the period of storage leading to the proliferation of microorganisms. This could be from aerosols or by chance inoculation, through handling or materials used or other microorganisms which are inherent in the vegetables because of their contact with the soil or water used in the washing of the leaves.

Bacterial Profile And Antibiotic Susceptibility Pattern Of Urinary Tract Infection Among Children Attending Felege Hiwot Referral Hospital, Bahir Dar, Northwest Ethiopia.

BackgroundUrinary tract infection (UTI) is a common and important clinical problem in pediatrics. Recurrent UTIs may lead to renal scarring, hypertension, and end-stage renal dysfunction later in life. The objective of the study was to determine bacterial profile and antimicrobial susceptibility pattern of urinary tract infections (UTIs) among children attending Felege Hiwot Referral Hospital (FHRH).MethodsA cross-sectional study was conducted from February 2013 to May 2013 among children 5–15 years of age with symptoms of UTI. Samples were processed for culture and identification. Antimicrobial susceptibility was done for positive urine cultures by the Kirby-Bauer’s disk diffusion method based on standards of the Clinical Laboratory Standard Institute (CLSI). Data were entered into Epi-data version 3.2.1 and exported to the Statistical Package for the Social Science (SPSS) version 20 statistical software. Fisher’s exact test and binary logistic regression test results were used.ResultsA total of 259 urine samples were collected from children with UTI. The result revealed 41 (15.8%) samples had significant bacteriuria, among which the most prevalent pathogen was E. coli 14 (34.1%) followed by Pseudomonas species. Gram-negative bacteria showed high level of sensitivity to ciprofloxacin (70), norfloxacin (63.4%) and ceftriaxone (60%), whereas the level of resistance was high to ampicillin (80%) and nitrofurantoin (70%). Gram-positive isolates showed high sensitivity to ciprofloxacin (77.8%), penicillin (72.8%) and erythromycin (72.7%). Multiple drug resistance (MDR) for Gram-positive and Gram-negative bacteria was 100% and 83.1%, respectively.ConclusionE. coli is the predominant bacteria isolated in the present study. The results showed that the prevalence of resistance to at least one antibiotic to commonly prescribed antimicrobials was high. Hence, the guidelines for empiric treatment of UTI should be re-evaluated periodically based on local studies.

Occurrence of Multi-drug Resistant Bacteria in Some Selected Street Food Samples

Antimicrobial resistance is a subject of great concern in the public health. The prevalence of antimicrobial resistance among food pathogens has increased during recent decades.
 Studying the incidence and antibiotic resistance pattern of bacterial species isolated from fish and vended street fruits. 
 Eleven fish swabs and thirteen sliced fruit samples were collected and prepared for isolation of bacterial species through inoculation onto selective and non-selective nutrient media. The grown colonies were purified through subculturing on nutrient agar plates then identified by morphological and biochemical methods. The obtained pure cultures were then kept on nutrient agar slants. Testing antibiotic resistance of the isolated bacterial strains was studied by Kirby-Bauer disk diffusion method on Mueller Hinton agar using ten antibiotics belonging to different classes. The resultant inhibition zone was interpreted according to Clinical Laboratory Standard Institute.
 Twenty-eight bacterial cultures were isolated from the collected food samples. The conventional identification using morphological and biochemical methods of these cultures revealed presence of three Gram positive species; Staphylococcus aureus, Streptococcus sp. and Bacillus subtilis in addition to four Gram negative; Escherichia coli, Brucella sp., Enterococcus faecalis and Proteus mirabilis. The incidence of the obtained bacterial species was arranged as 29.16% for both S. aureus and E. faecalis followed by Brucella sp. 16.66%; B. subtilis & E. coli 12.5% then Streptococcus sp. and P. mirabilis with an incidence of 8.33% each. Testing antibiotic resistance pattern of seven bacterial species against ten antibiotics showed that, among three Gram positive bacterial species, only one (33.33%) strain S. aureus exhibited resistance to six antibiotics; amoxicillin, erythromycin, ciprofloxacin, ceftriaxone, fluconazole and dicloxacillin. Among four Gram negative bacterial strains only one (25.0%) strain Enterococcus faecalis exhibited resistance to eight antibiotics; amoxicillin, streptomycin, chloramphenicol, cotrimoxazole, ciprofloxacin, ofloxacin, sparfloxacin and cloxacillin.
 Occurrence of multi-drug resistant bacteria in fish and vended street fruits poses not only risk of disease to the foods but public health hazard to food handlers and consumers in general. Also the result of this study recommended augmentin and cephazolin as good choice antibiotics for treatment of infection in the study area.

1608. Use of Selective Reporting of Antimicrobial Susceptibilities and Its Impact on Antimicrobial Resistance Surveillance—National Healthcare Safety Network, 2017–2018

BackgroundSelective reporting (SR), recommended by the 2016 IDSA/SHEA antimicrobial stewardship guidelines, is a strategy to guide prescribing decisions by limiting the antimicrobial susceptibility testing (AST) results available to prescribers. Yet, SR carries risks that cumulative antibiograms reflect only partial AST results. The Clinical Laboratory Standards Institute (CLSI) M100 performance standards stipulate that AST results should be routinely reported for some antimicrobials (Group A agents) while SR is appropriate for other antimicrobials (Group B agents). We assessed the extent of SR use and its impact on national antimicrobial resistance (AR) surveillance.MethodsWe used Enterobacteriaceae (EB) and Staphylococcus aureus (SA) blood culture AST results that hospitals reported for group A and B agents to the CDC’s National Healthcare Safety Network’s AR option from 2017 through 2018. Routine reporting for an organism-agent combination was defined as results reported for ≥ 90% isolates for the hospital’s most frequently reported agents. SR was defined as a shortfall of > 20% in results reported for an agent compared with a routinely reported agent in a hospital that reported ≥ 30 isolates. We compared hospital antibiograms between SR and non-SR hospitals. We also identified isolate characteristics associated with AST reporting in SR hospitals.ResultsAmong 242 and 185 hospitals reported ≥ 30 isolates, many showed patterns of SR (Figure 1). Of 437 and 425 hospitals reported ≥ 1 isolate, only 112 (26%) and 152 (36%) routinely reported AST results for all group A agents for EB and SA, respectively. For EB, 345 (79%) hospitals routinely reported AST results for ciprofloxacin or levofloxacin, although both are group B agents. For SA, 324 (76%) routinely reported vancomycin (Figure 2). Antibiograms for many agents differed between SR and non-SR hospitals (Figure 3, 4). In SR hospitals, non-susceptibility to narrower-spectrum drugs, patient location, age, and some species among EB were associated with AST reporting.ConclusionAST results reporting vary across hospitals and agents, and CLSI’s SR standards are used inconsistently. For AR surveillance, complete reporting calls for solutions that bypass SR. In the meantime, SR should be taken into account in national AR benchmarking. Disclosures All authors: No reported disclosures.

528. Longitudinal Evaluation of Chlorhexidine Resistance Among Multidrug-Resistant Organisms in Relation to Intervention of Daily Chlorhexidine Bathing in Adult Intensive Care Units

BackgroundChlorhexidine digluconate (CHG), the most widely used antiseptic, has recently been applied to patient washing to decolonize the multidrug-resistant organisms (MDROs), but there are little data on susceptibilities of MDROs to CHG. The purpose of this study was to evaluate CHG resistance among MDROs before and after the intervention of daily CHG bathing in adult intensive care units (ICUs).MethodsThe intervention of daily body washing with 2% CHG cloths were taken in adult patients the medical or surgical ICU of 23-bed by a crossover manner for 6 months (MICU, July to December 2017; SICU, January to June 2018) in a 1,050-bed, university hospital in the Republic of Korea. Available MDRO isolates were randomly selected from clinical cultures of ICU patients within 6 months before, during and after the intervention, including MRSA, MR-CoNS, VRE, Carbapenem-resistant Pseudomonas aeruginosa (CR-PA), CR-Acinetobacter baumannii (CR-AB). Minimum inhibitory concentrations (MICs) were determined using the broth microdilution method set by the Clinical Laboratory Standards Institute. Determination of the minimum bactericidal concentrations (MBCs) was performed by subculturing 10 µL from each well without visible microbial growth. Cumulative amounts of CHG used in both ICUs was estimated across the study period from January 2008 to June 2018.ResultsThe cumulative CHG consumption from both ICUs increased sharply from 27,503 g to 29,556 g after one-year intervention. The ranges of MICs and MBCs of CHG among MDRO clinical isolates selected by a 6-month phase are summarized in Table 1. Particularly, CR-PA and CR-AB isolates revealed four to eight times higher MICs and MBCs compared with the majority of Gram-positives excepting some VRE isolates. On the other hand, neither MICs and MBCs ranges of CHG from the MDRO isolates nor the monthly incidence of the MDROs from both ICUs were significantly increased before and after the intervention of daily CHG bathing.ConclusionThis study indicates that some Gram-negative MDRO isolates with higher MICs and MBCs of CHG might be from longstanding exposure to CHG or efflux pumps. Although 2% daily CHG bathing uses over 1,000 times higher concentrations than the lethal concentration, it might be needed to monitor CHG resistance among MDROs. Disclosures All authors: No reported disclosures.

1467. Antimicrobial Susceptibility and Molecular Characterization of Extended-Spectrum β-Lactamase of Escherichia coli and Klebsiella pneumoniae of Urine Samples Isolated from Community Patients in South Brazil

BackgroundEnterobacteriaceae is the main pathogens of UTI. It is important to be aware the local epidemiological data for an appropriate initial treatment. Resistance to antimicrobial agents has increased, especially to first-choice antibiotics in the treatment of cystitis. Our objective is to asses the antimicrobial susceptibility profile from uropathogens isolated in community and evaluated the dissemination of extended-spectrum ß lactamase (ESBL), in E. coli and K. pneumoniae in south of Brazil.MethodsFrom June 2016 to June 2017, all urine samples collected in the Basic Health Units and Emergency Departments were sent to a Central Laboratory. Identification and susceptibility tests were performed on the VITEK® 2 (bioMérieux, France) system. Clinical Laboratory Standards Institute (CLSI) breakpoints were used for the interpretation of susceptibility. Positive cultures were defined as those demonstrating ≥105 CFU / mL (colony-forming units). The presence of ESBL was also subjected to the Chrom ID BLEE® agar plate test (bioMérieux- Marcyl’Etoile, France). PCR technique uses specific primers for genes blaTEM and blaSHV. Detection of the blaCTX-M genes was performed by multiplex PCR.ResultsA total of 56,555 microbiologic tests were performed, 8189 were positive. Women were responsible for 89.4%, and 10% were pregnant. Table 1 shows uropathogens isolated. Graphic 1 shows antimicrobial susceptibility. Extended-spectrum ß lactamase production was present in 6.7% (n = 489). People older than 60 years had ESBL more frequent (P <0.05) as well as being pregnant is not related to ESBL (P <0.05). Table 2 shows the distribution of the bla genotypes. Table 3: Distribution of blaCTX-M. Among blaCTX-M1 genotype, blaCTX-M15 was the most frequent.ConclusionIn this study, the most frequent uropathogen isolated was E. coli followed by K. pneumoniae. Cotrimoxazol had high rates of resistance and nitrofurantoin the lowest. Quinolone resistance is more than 10%. Sensitivity to aminoglycosides and carbapenems remains high.We found relevant frequency of ESBL, CTX-M-1-group most commonly found. Among CTX-M-1, blaCTX-M15 was the most isolated. Disclosures All authors: No reported disclosures.

634. Incidence of vanC-Mediated Vancomycin-Resistant Enterococcus Bloodstream Infections at Children’s Hospital of Colorado and Implications for Empiric Enterococcal Therapy

BackgroundThe most well-known Enterococcal species, E. faecium and E. faecalis, can harbor high-level vancomycin resistance mediated by acquired vanA and vanB operons. However, other Enterococcal species such as E. gallinarum and E. casseliflavus (VCE), harbor intrinsic low-level vancomycin resistance mediated by an intrinsic vanC operon, and the incidence of these pathogens among pediatric patients is not clear. As the antibiotic resistance pattern of VCE is different than E. faecium and E. faecalis, a high prevalence of VCE may have implications for antibiotic therapy. We describe the incidence and susceptibility of VCE bloodstream infections at a large children’s hospital and compare to E. faecalis and E. faecium.MethodsPositive blood culture results from 2013 to 2018 were obtained from the Children’s Hospital of Colorado data warehouse. All first-time positive cultures for Enterococcus were analyzed for species, susceptibility, and hospital unit location. First-time positive was defined as being at least 2 weeks after any previous positive Enterococcus blood culture. Susceptibilities were categorized by clinical laboratories standards institute (CLSI) guidelines.ResultsOf 240 positive isolates, 7% were ampicillin susceptible and vancomycin nonsusceptible (resistant or intermediate), vs. 6% that were ampicillin resistant and vancomycin susceptible. An additional 3% of isolates were not susceptible to either antibiotic; all of these were E. faecium. VCE accounted for 12% of our isolates while E. faecalis and E. faecium accounted for 66% and 16%, respectively. All VCE were susceptible to ampicillin, but 52% were nonsusceptible to vancomycin. VCE incidence, ampicillin resistance, and vancomycin nonsusceptibility were most prevalent in our hematology, oncology, and bone marrow transplant (BMT) units.ConclusionAt our institution, an as yet unspeciated Enterococcus is equally likely to be ampicillin susceptible and vancomycin nonsusceptible as ampicillin resistant and vancomycin susceptible. This is driven by a significant incidence of VCE, especially on our hematology, oncology, and BMT units. Therefore, vancomycin may not provide adequate empiric Enterococcal coverage on these units, and the addition of ampicillin will be recommended. Disclosures All authors: No reported disclosures.

2030. Impact of an Antimicrobial Stewardship Program on Carbapenem Susceptibility in a National Hospital in Bhutan

BackgroundThe overuse of broad-spectrum antibiotics drives antimicrobial resistance (AMR), and the prevalence of highly-resistant Gram-negative infections is increasing across the world, especially in low- and middle-income countries (LMIC). Carbapenem resistance is of particular concern since these are often the last line agents. Antimicrobial restriction is an antimicrobial stewardship intervention (AMS) that aims to reduce the use of broad-spectrum antibiotics to preserve antimicrobial susceptibility.MethodsThis is retrospective, observational study of antibiotic consumption and prevalence of antibiotic resistance of bacterial isolates from inpatients at Jigme Dorji Wangchuck National Referral Hospital, a 350-bed multi-specialty hospital in Thimphu, Bhutan. Antibiotic consumption and antimicrobial susceptibility were monitored from January 2015 to December 2017 by the pharmacy department and the microbiology lab, respectively. Antibiotic consumption was measured using defined daily doses (DDD) and expressed as DDDs per 1,000 persons per day. The antibiotic susceptibility was determined using the Clinical Laboratory Standards Institute (CLSI) guideline. A hospital AMS program with multidisciplinary team and good hospital managerial/ leadership support were initiated in 2016 and interventions included antimicrobial restrictions, educations, guidelines for use, post prescription review, de-escalation, audit and feedback.ResultsFrom 2015 to 2016, the DDDs of carbapenems and piperacillin–tazobactam (PTZ) increased while ceftriaxone decreased (Figure 1). After the AMS program was implemented in 2016, the annual DDDs of carbapenems decreased while PTZ and ceftriaxone increased. Antimicrobial susceptibility of Klebsiella pneumoniae and Escheriachia coli blood isolates to carbapenems and ceftriaxone increased from 2016 to 2017: 50/61 (82%) vs. 45/49 (92%) and 24/91 (26%) vs. 31/92 (34%), respectively.ConclusionImplementing an AMS program that restricted the use of carbapenems resulted in a decrease in carbapenem use and increased antimicrobial susceptibility for carbapenems and ceftriaxone. AMS interventions can be successful to decrease carbapenem-resistance in LMIC.

1594. Ceftolozane–Tazobactam Demonstrates Higher In Vitro Susceptibility than Ceftazidime–Avibactam Against Pseudomonas aeruginosa Isolated from Respiratory Tract of Adult Cystic Fibrosis Patients

Background Pseudomonas aeruginosa is a commonly isolated pathogen in adults with cystic fibrosis (CF). Antimicrobial resistance is an escalating problem due to chronic colonization and frequent antimicrobial exposure. Ceftolozane–tazobactam (C/T) and ceftazidime–avibactam (CZA) exhibit promising activity against antimicrobial-resistant organisms, including P. aeruginosa. In this study, we compared in vitro activity of C/T and CZA against P. aeruginosa isolated from respiratory cultures obtained from adult patients with CF.MethodsThis is a retrospective study of respiratory cultures positive for P. aeruginosa collected from adult CF patients between January 1, 2015 to November 30, 2018. The first isolate per patient per year that underwent susceptibility testing for C/T, CZA, and colistin were included in the study. All isolates underwent in-house susceptibility testing for 9 anti-pseudomonal agents according to the methodology established by the Clinical Laboratory Standards Institute (CLSI). Susceptibility testing of C/T, CZA, and colistin were performed by a reference lab. Isolates were classified into 3 drug-resistant categories using the following definition: multidrug-resistant (MDR) non-susceptible (NS) to ≥1 agent in ≥3 different antimicrobial classes, extensive drug-resistant (XDR) NS to 4 or 5 different classes, and pan drug-resistant (PDR) NS to all 6 classes except colistin.ResultsForty-two P. aeruginosa respiratory isolates from 32 patients with CF were included. The overall susceptibility to C/T and CZA was 59.5% and 42.9%, respectively. Thirty-eight (90%) isolates were considered MDR with susceptibility of 55.3% to C/T and 44.7% to CZA. Among the 11 XDR isolates, susceptibility to C/T was 81.8% vs. CZA 72.7%. Susceptibility to C/T vs. CZA was also higher (37.5% vs. 25%) among the 24 PDR isolates.ConclusionAmong P. aeruginosa isolated from CF respiratory cultures, C/T appears to have better in vitro activity compared with CZA, and remained true among isolates considered XDR and PDR. These results suggest using C/T while awaiting susceptibilities when standard anti-pseudomonal agents cannot be used. Future studies evaluating clinical outcomes for the treatment of pulmonary CF exacerbations are needed to assess the applicability of in vitro susceptibility data.Disclosures All authors: No reported disclosures.

1955. Novel Compound Reverses Vancomycin Resistance in Vancomycin-resistant Enterococci (VRE)

Background Enterococcus causes 14% of all hospital-associated infections (HAIs) according to Centers for Disease Control and Prevention (CDC) data. 35.5% of these HAIs are caused by Vancomycin-resistant Enterococci (VRE) including highly fatal bacteremia, surgical site infections, and urinary tract infections. We present a novel synthetic compound, HSD 03-21 that could make VRE completely susceptible to vancomycin in-vitro.MethodsHSD 03-21 was synthesized de novo from a hydroxybenzylidene – indolinone backbone in our laboratory. The minimum inhibitory concentration (MIC) of HSD 03-21 and vancomycin against VRE were determined according to clinical laboratory standards institute (CLSI) guidelines. The standard checkerboard assay was used to determine vancomycin-HSD 03-21 interactions against VRE. Briefly, HSD 03-21 and vancomycin at 10 mg/mL were prepared and diluted serially along the ordinate and abscissa of 96-well microtiter plates, respectively. Bacteria was standardized using the 0.5 McFarland standard, diluted (1:100), aliquoted into respective wells and incubated at 37oC for 18–20 hours. All assays were run in triplicates. The fractional inhibitory concentration (FIC) index was calculated for each combination. The FIC of either agent was calculated as: FIC (vancomycin) = MIC of vancomycin in combination/MIC of vancomycin alone and FIC (HSD 03-21) = MIC of HSD 03-21 in combination/MIC of HSD 03-21 alone. The cumulative FIC index ∑FICI was then calculated as: ∑FIC = FIC(vancomycin) + FIC(HSD 03-21). The calculated ∑FIC indices were interpreted as synergistic if ∑FIC: ≤ 0.5.ResultsThe MIC of vancomycin for VRE faecalis was 256 μg mL−1 while that of HSD 03-21 was 128 μg mL−1. When vancomycin was combined with HSD 03-21 at 8 μg mL−1 (1/16 MIC), there was a reduction in MIC of vancomycin to 0.5 μg mL−1. The combination showed excellent synergy with ∑FIC of 0.06.ConclusionHSD 03-21 reduced the MIC of vancomycin from 256 to 0.5 μg mL−1. This has an immense potential of changing the way we use vancomycin and in the treatment of VRE infections. Translation of this novel compound could save thousands of lives from VRE and the failures and inherent toxicities of current doses of vancomycin. Disclosures All Authors: No reported Disclosures.

630. Clinical and Molecular Characteristics of Carbapenem-Resistant Enterobacteriaceae in Qatar: A Retrospective and Prospective Observational Study

BackgroundThere are limited data describing carbapenem-resistant Enterobacteriaceae (CRE) from the Arabian Gulf region. Our aims were to describe the clinical, microbiological and molecular characteristics of CRE infections and to characterize the gene displaying resistance to carbapenem.MethodsRetrospective and prospective review of clinical, microbiological and molecular characteristics of CRE isolated between April 2015 and November 2017 at 7 tertiary hospitals in Qatar. Susceptibility testing was performed and interpreted according to Clinical Laboratory Standards Institute methodology and breakpoints. Carbapenemase genes were identified using PCR. Whole-genome sequencing followed by bioinformatics analysis was performed on a subset of samples for accurate detection of resistant genes. SPSS V.21.0 was used for all statistical analyses.ResultsA total of 144 nonrepeating CRE strains were recovered from 137 individuals over the study period, of which 35 (24.3%) represented colonization. Klebsiella spp. (98, 68.1%) and E. coli (38, 26.4%) were the most predominant. The most prevalent carbapenemases were NDM (69, 47.9%), OXA-48 (36, 25.0%), and KPC (15, 10.4%). Sixteen (11.1%) strains possessed multiple carbapenemase genes but none were detected in 37 (25.7%) strains. Genomic analysis also detected the presence of NDM-7, NDM-5, NDM1, VIM4, and OXA48-like carbapenemase genes among 34 clinical samples. The majority of the strains were susceptible to tigecycline (124, 86.1%), fosfomycin (122, 84.7%) and amikacin (102, 70.8%). There were 109 CRE infections involving the urinary tract (40, 36.7%), bloodstream (31, 28.4%), and respiratory tract (17, 15.6%). Fifty-seven (52.3%) were males and the median age was 57 years (range 3–97). Baseline comorbidities included diabetes (57, 52.3%), chronic kidney disease (27, 24.8%), and cancer (27, 24.8%) (Table 1). Within the 30 days preceding CRE infection, 38 (34.9%) had received carbapenem therapy, 65 (59.6%) had undergone invasive procedures and 33 (30.3%) had had surgery. All-cause 30-day mortality was 29.4%. The only variable independently associated with mortality was baseline SAPS Score (Table 2).ConclusionNDM and OXA-48 are the predominant carbapenemases in Qatar. CRE infections are associated with high overall mortality. Disclosures All authors: No reported disclosures.

Multi-site performance evaluation and Sigma metrics of 20 assays on the Atellica chemistry and immunoassay analyzers.

Background The Atellica Solution comprises chemistry (CH) and immunoassay (IM) analyzers. Recently, six early adopter clinical laboratories across Europe evaluated the analytical performance of 20 CH and IM assays. To measure analytical performance quality, Sigma metrics were calculated for individual-site and pooled-site results. Methods Precision, detection capability, linearity, and method comparison studies were performed according to Clinical Laboratory Standards Institute protocols. Global Sigma metrics across sites were calculated from pooled data at the medical decision level using total allowable error (TEa) goals from CLIA for CH assays, and TEa goals from RiliBÄK for IM assays; and, the equation: Sigma metrics=%TEa-%bias/%CV. A pooled %CV was calculated by combining the imprecision obtained from individual sites. Bias calculations were performed against the ADVIA Chemistry system or ADVIA Centaur system using Deming regression analysis (Passing-Bablok regression for electrolytes) on the pooled-site data. The 103 individual-site Sigma metric calculations used individual-site imprecision and pooled-bias. Results The limits of blank and detection results agreed with the manufacturer's claims. Most assays were linear across the assay range tested. Pooled Sigma metrics were good or better (>4 Sigma) for 18 of 20 assays; and, acceptable for urea nitrogen (3.1) and sodium (3.9), the latter values attributable to higher imprecision at one of five sites. Conclusions Sigma metrics for data generated across multiple real-world sites evaluating the Atellica Solution demonstrated good or better performance of greater than 4 Sigma for 18 of 20 assays tested. Overall, results verified the manufacturer's claims that methods were fit for use in clinical laboratories.

Antibiotic resistance pattern of methicillin-resistant &lt;i&gt;Staphylococcus aureus&lt;/i&gt; isolated from children under the age of five Years in Anambra State

Neonates are exposed to Staphylococcus aureus shortly after birth and can become colonized quickly after contact with adult skin or the environment. Methicillin-resistant S. aureus (MRSA) constitutes part of the growing global health problem associated with an increasing number of infections and often multidrug resistant in nature which now poses serious therapeutic problems to clinicians. Eighty-three (83) samples were collected which were cultured on appropriate bacteriological media. Bacterial isolates ( S. aureus ) were identified by standard biochemical tests and confirmed using PCR targeting the 16S rRNA. The MRSA was determined using Oxacillin antibiotic disk and confirmed by the presence of MecA gene. Antibiotic susceptibility of the MRSA isolates to eleven antibiotics was performed according to Clinical Laboratory Standard testing Institute (CLSI) guidelines. Out of the 83 individuals tested, 25 yielded S. aureus of which 22 (88.0%) of them were MRSA positive, 10 (45.5%) females and 12 (54.5%) males. The antibiotic resistant pattern of the 22 MRSA isolates showed Quinupristin/Dalfopristin 13 (59.1%), Fusidic acid 6 (27.3%), Linezolid 8 (36.4%), Clindamycin 10 (45.5%), Vancomycin 5(22.7%), Cefepime 3(13.6%), Doxycycline 4(18.2%), Sulphamethoxazole 22(100%), Fosfomycin 7(31.8%), Cephalexin 7(31.8%), Trimethoprim 19(86.4%). The MIC determination for vancomycin from MRSA isolates showed antibiotic concentration of 4.9 μg/ml and 8.7 μg/ml. This work showed that there is high prevalence rate of resistance to many classes of antibiotic warranting continued surveillance and antimicrobial stewardship. Therefore effective antibiotic susceptibility test should be conducted before prescribing an antibiotic to patients, in as much as patients should strictly adhere to antibiotic prescription to mitigate abuse of drugs. Keywords: Methicillin-resistance, antibiotics, prevalence, S. aureus , susceptibility

To study the profile of Candida isolates and antifungal susceptibility pattern of neonatal sepsis in a tertiary care hospital of North India

Aims and objective To study the profile of Candida infection and antifungal susceptibility (AFS) pattern in neonatal sepsis. Materials and methods In this prospective observational study, 850 blood samples were collected and processed from neonates who were suspected clinically to have sepsis. The blood culture that showed growth of Candida was further processed for species identification. Antifungal susceptibility was done as per the National Committee for Clinical Laboratory Standards (NCCLS)/Clinical Laboratory Standards Institute (CLSI) M44-A-2 guidelines by disc diffusion method and automated vitek-2 compact system. Candidemia was diagnosed by isolation of Candida species from at least one positive blood culture containing pure growth of Candida species with supportive clinical features. Results Candida species were isolated in 32 specimens of the total 322 culture-positive cases. The most common isolate was Candida tropicalis (14/32; 43.75%) followed by Candida albicans (7/32; 21.87%) and Candida glabrata (6/32; 18.75%). The three most common neonatal risk factors for candidemia were low birth weight, prolonged use of intravenous antibiotics and presence of central venous line. Non-albicans Candida (NAC) showed good sensitivity to fluconazole as compared to Candida albicans. The fluconazole sensitivity of Candida tropicalis, Candida glabrata, and Candida parapsilosis was 93, 67, and 100%, respectively, whereas it was 57% in Candida albicans. The sensitivity to amphotericin B was 95% among all Candida isolates. All NAC were sensitive to amphotericin B, while only 72% Candida albicans were sensitive to amphotericin B. Conclusions The incidence of neonatal sepsis secondary to NAC is increasing and has replaced Candida albicans as a major cause of neonatal fungal sepsis. Low birth weight is the most important risk factor for Candida sepsis. The resistance of Candida albicans is increasing for both fluconazole and amphotericin B when compared to NAC. Increasing antifungal resistance warrants its judicious use both for prophylaxis and treatment.

P3599Minicare high sensitivity troponin: a novel point-of-care tool to improve ACS workflows

Abstract Background Cardiac troponin (cTn) has been broadly accepted as the biomarker of choice in the diagnosis of patients presenting with Acute Coronary Syndrome (ACS). Point-of-care (POC) cTn testing, defined as testing near the patient with rapid availability of results, has attracted a strong interest in the emergency department (ED) setting. It offers the potential to improve workflows, expedite clinical decisions and to reduce the length of stay. Workflows could be further optimized when a first measurement can already be performed by the emergency medical services prior to admission to the hospital. Advances in assay technology have led to high sensitive (HS) cTn assays that have a profound impact on clinical practice, providing early results at presentation and 1 hour after admission which speed up clinical decision-making. Here we evaluate the Minicare HS cTnI POC test under development, which has the potential to combine the benefits of HS-cTnI protocols with a POC workflow. Objective Evaluate the analytical and clinical capability of the Minicare HS-cTnI test under development to meet the criteria‡ for HS cTn of having a 10% CV &lt;99th percentile and a percentage measurable (&gt;LoD) of &gt;50%. Methods The evaluation is based on the Clinical Laboratory Standards Institute (CLSI) guidelines. Li-heparin whole blood and Li-heparin plasma samples were used to establish LoB, LoD, and LoQ, sample matrix comparison and linearity in the low range. The clinical performance for Minicare HS-cTnI was assessed and compared to Abbott Architect high-sensitivity troponin using banked samples from patients with ACS and collected at t=2–4h after admission. The percentage measurable was assessed and compared to Abbott Architect high-sensitivity troponin in a healthy population of 165 individuals. Results With a time to result within 10 minutes, the LoB is determined at 1.0 ng/L and the LoD at 2.1 ng/L. Plasma and whole blood results correlated well and showed comparable results. The 10% and 20% CV LoQ are established at 7.9 ng/L and 3.1 ng/L, well below the anticipated 99th percentile of 26 ng/L. With a percentage measurable of 77%, the criteria for a HS-cTnI assay are met in a POC device. A sensitivity of 93% and an NPV of 99% are found for Minicare HS-cTnI for a t=2–4h protocol vs 90% and 98% for Abbott Architect high-sensitivity troponin. In this study the area under the curve is 0.98 for Minicare HS-cTnI and 0.97 for Architect HS-cTnI. ROC curve t=2-4 h Conclusions With HS-cTnI capability on the Minicare platform, we show the potential to support a 0/1 h sampling protocol, combined with the speed of a POC workflow. This may enable even more rapid and safe rule-out of patients with ACS.

Prevalence and Susceptibility Analysis of Gram Negative Pathogens in Super Specialty Tertiary Care Centers, Pune in India from January 2018- January 2019

Background and Objective: The encumbrance of antimicrobial resistance worldwide is substantive and likely to rise without any appropriate treatment which has enhanced the urge for the development of either new antibiotics or adjuvant therapy with antibiotics. Thus, we aimed to study a comparative antibiogram pattern of 758 clinical isolates collected from Pawana Hospital and Accord SDH, Pune (India), towards Elores (a novel antibiotic and adjutant entity of ceftriaxone, sulbactam and disodium edetate) and other antibiotics (imipenem, meropenem and piperacillin + tazobactam).&#x0D; Methods: The clinical samples collected from outpatients and inpatients during a period of one year (January, 2018 to January, 2019), from Pawana Hospital and Accord SDH, Pune (India) and were further subjected to bacterial identification. Antibiotic susceptibility testing was executed in accordance with the recommendations of Clinical Laboratory Standards Institute (CLSI) guidelines.&#x0D; Results: Out of 758 collected samples, urine samples contributed 69.41 and 37.63% among Enterobacteriaceae and Non-Enterobacteriaceae followed by pus (20.25 and 19.35%) and sputum (4.2 and 19.89%) while &lt;4% and &lt;12% Enterobacteriaceae and Non-Enterobacteriaceae isolates were collected from rest of the specimen. E. coli were found most prevalent (50.13%) along with 19.13 and 16.09% prevalence of Pseudomonas spp. and Klebsiella spp. whereas rest of the pathogens were &lt;7% present. The antibacterial activity of Elores (87.80%) was observed superior to carbapenem drugs (meropenem; 62.67% and imipenem; 60.95%) and far better to piperacillin + tazobactam (48.42%) against 758 clinical pathogens. Antibiogram profile depicted Elores as most susceptible (95.10%) drug towards Enterobacteriaceae isolates which was approximately 31-50% more sensitive than other test drugs. Similar pattern was obtained for Non-enterobacteriaceae isolates (62.90%) where Elores contributed approximately 1-5% higher activity. However, Elores also conquered 79.51-86.59% resistance among meropenem, imipenem and piperacillin + tazobactam resistant pathogens.&#x0D; Conclusion: Susceptibility profile data revealed the equivalence of Elores (Antibiotic-adjuvant entity; AAE) with carbapenem drugs (meropenem and imipenem) and superiority over piperacillin + tazobactam against clinical pathogens. Elores was also found active towards meropenem, imipenem and piperacillin + tazobactam resistant pathogens. Therefore, Elores, a resistance breaker, can be used as an efficient treatment alternate towards infections caused by resistant pathogens.

Bacteriological Profile of Wound Swab and Their Antimicrobial Susceptibility Pattern in Shaheed Suhrawardy Medical College, Dhaka

Background: Wound infections are global problem in the field of surgery associated with long hospital stay, higher treatment expenditure, morbidity and mortality.&#x0D; Objective: To isolate and identify the bacteria causing wound infection and to determine the antimicrobial susceptibility pattern.&#x0D; Methods: This retrospective study was conducted in the Department of Microbiology at Shaheed Suhrawady Medical College, Dhaka from January 2017 to December 2017 for a period of one (01) year. A total of 190 wound swabs were collected from the patients who were visited in outpatient department and were admitted at inpatient department with skin and soft tissue infection. Swabs from the wound were inoculated on appropriate media and cultured and the isolates were identified by standard procedures as needed. Antimicrobial susceptibility testing was done by disc diffusion method according to ‘The Clinical Laboratory Standard Institute’ guidelines.&#x0D; Results: In this study, out of 190 cases 115 (60.52%) were male and 75 (39.47%) were female and majority 85(44.73%) were in the age group of 16 to 30 years. A total number of 190 isolates were obtained, among which 124 (65.25%) were culture positive cases. Among the isolated organisms predominant bacteria was Staphylococcus aureus 68 (35.79%) followed by Escherichia coli 30 (15.79%), Pseudomonas 14 (7.37%), Klebsiella 6 (3.16%), Proteus 4 (2.10%) and Acinetobacter 2 (1.05%). Staphylococcus aureus was sensitive to linezolid (94.11%), vancomycin (88.23%) and amikacin (70.58%). Among the Gram negative isolates Escherichia coli was predominant and showed sensitivity to imipenem (80%), amikacin (70%), ceftazidime (60%), piperacillin+ tazobactum (56.66%), colistin (53.33%). Pseudomonas showed sensitivity to colistin (78.57%), imipenem (71.42%). Klebsiella showed sensitivity to imipenem (83.33%), amikacin (66.66%), piperacillin+ tazobactum (66.66%), and colistin(66.66%).&#x0D; Conclusion: Staphylococcus aureus was the most frequently isolated pathogen from wound swab and the antibiotic sensitivity pattern of various isolates will guide for appropriate selection of antibiotics against wound infection, so as to reduce the spread of resistant bacteria.&#x0D; J Shaheed Suhrawardy Med Coll, June 2019, Vol.11(1); 65-68

Resistencia a la meticilina y producción de biopelícula en aislamientos clínicos de Staphylococcus aureus y Staphylococcus coagulasa negativa en México

ResumenIntroducción. Las infecciones por Staphylococcus aureus y Staphylococcus coagulasa negativa multirresistentes a los antibióticos y asociadas con la atención en salud tienen un gran impacto epidemiológico por su alta morbimortalidad; además, se han relacionado con la formación de biopelículas, lo cual también se asocia con la resistencia a los antimicrobianos.Objetivo. Determinar la resistencia a la meticilina y cuantificar la producción de biopelículas para establecer su posible relación con los aislamientos clínicos de S. aureus y Staphylococcus coagulasa negativa.Materiales y métodos. Se estudiaron 11 cepas de S. aureus y 12 de Staphylococcus coagulasa negativa. La resistencia a la meticilina se determinó con discos de cefoxitina tomando como valores de referencia los estándares del Clinical Laboratory Standards Institute (CLSI) de 2018. La producción de biopelícula se cuantificó con cristal violeta. Los genes mecA e icaADBC se identificaron mediante reacción en cadena de la polimerasa (PCR), y se hizo un análisis bivariado con la prueba de ji al cuadrado y el coeficiente V de Cramér, utilizando el programa SPSS™, versión 20.0.Resultados. Nueve cepas de S. aureus fueron resistentes a la meticilina (SARM) y dos fueron sensibles. Ocho cepas de Staphylococcus coagulasa negativa fueron resistentes y cuatro fueron sensibles. El genotipo mecA se encontró en ocho de las nueve cepas de S. aureus y en seis de las ocho de Staphylococcus coagulasa negativa resistentes a meticilina. Todas las cepas formaron biopelícula. Diez cepas de S. aureus y 11 de Staphylococcus coagulasa negativa presentaron el genotipo icaADCB. No se encontró asociación entre la resistencia a meticilina y la formación de biopelícula.Conclusiones. La cefoxitina es suficiente para determinar el fenotipo resistente a meticilina y se asoció con el genotipo mecA. Las cepas resistentes a la meticilina y poseedoras del gen mecA pueden presentar un mecanismo de resistencia alterno. Los dos grupos de cepas formadoras de biopelícula se relacionaron con la presencia del operón icaADCB. La formación de biopelícula y la resistencia a la meticilina se expresaron como características independientes en los dos grupos de cepas.

English

Antimicrobials have been playing an important role in preventing illness and death associated with infections due to bacteria. However, the emergence and spread of resistance by pathogens have decreased the effectiveness of the commonly prescribed antimicrobials. Intestinal Escherichia coli are among bacterial pathogens that are endowed with such resistance traits because they are important source and reservoir of genes that encode antimicrobial resistance. To determine the antimicrobial resistance profile of fecal isolates of E. coli from diarrheic patients. Stool samples were collected consecutively from 100 individuals who visited Selam Health Center during the study period, April to June 2018. Samples were collected and transported under sterile condition to the National Clinical Bacteriology and Mycology reference Laboratory, Ethiopian Public Health Institute. The samples were streaked on MacConkey agar and incubated overnight at 37°C. E. coli isolates were further confirmed using conventional biochemical tests. Antimicrobial susceptibility status was determined using the disk diffusion method on Mueller Hinton agar as recommended by the Clinical Laboratory Standard Institute. The raw data was compiled and entered to spreadsheet and analysis was done using SPSS Version 20 with p-value ≤0.05 considered statistically significant. Out of the 100 patients, 43 were female and the rest were male. Confirmed E. coli were isolated from 73 individuals. Antimicrobial susceptibility testing showed that E. coli isolated in this study were highly resistant to trimethoprim-sulfamethoxazole 49 (67.1%) and amoxicillin-clavulanic acid 47(64.4 %). No isolates showed resistance to gentamicin and tobramicin. Of all the isolates, 11(15.1%) were multidrug resistant. No association was observed between antimicrobial resistance status and sex of individuals included in this study. However, there was an association between age and resistance patterns. Resistance to commonly prescribed antibiotics among E. coli isolated in this study was high and a considerable proportions of the strains were multidrug resistant. This is an indication for an alarming rate of resistance of intestinal E. coli to first line antimicrobials. To reduce the problem, regular monitoring and education for the community are very important. Key words: Escherichia coli, antibiotic susceptibility, multidrug resistant, Ethiopia, biochemical tests, disk diffusion.

Establishing thresholds and effects of gender, age, and season for thyroglobulin and thyroid peroxidase antibodies by mining real-world big data

BackgroundThyroglobulin antibody (TG-Ab) and thyroid peroxidase antibody (TPO-Ab) are cornerstone biomarkers for autoimmune thyroid diseases, and establishment of appropriate thresholds is crucial for physicians to appropriately interpret test results. Therefore, we established the thresholds of TG-Ab and TPO-Ab in the Chinese population through analysis of real-world big data, and explored the influence of age, gender, and seasonal factors on their levels. MethodsThe data of 35,869 subjects downloaded from electronic health records were analyzed after filtering based on exclusion criteria and outliers. The influence of each factor on antibody levels was analyzed by stratification. Thresholds of TG-Ab and TPO-Ab were established through Clinical Laboratory Standards Institute document C28-A3 and National Academy of Clinical Biochemistry (NACB) guidelines, respectively. ResultsThere were significant differences according to gender after age stratification; the level of TG-Ab gradually increased with age in females. There were significant differences in TG-Ab and TPO-Ab distributions with respect to age after gender stratification. Moreover, differences were observed between seasons for TG-Ab and TPO-Ab. The thresholds of TG-Ab and TPO-Ab were 107 [90% confidence interval (CI):101–115] IU/mL and 29 (90% CI: 28–30) IU/mL, respectively, using C28-A3 guidelines, but were 84 (90% l CI: 50–126) IU/mL and 29 (90% CI: 27–34) IU/mL, respectively, using NACB guidelines. ConclusionThe levels of TG-Ab and TPO-Ab were significantly affected by gender, age, and season. The thresholds for TG-Ab and TPO-Ab for the Chinese population were established by big data analysis.