Frequency and Antimicrobial Susceptibility Pattern of Gram-Negative Bacilli Isolated From Urine Specimens at a Tertiary Care Setting

OBJECTIVES: To find out the frequency and pattern of conventional antibiotic susceptibility of gram-negative bacilli cultured from urine specimens of patients at a tertiary care setting. METHODOLOGY: This study was conducted at the Microbiology Department of Combined Military Hospital Multan from June 2016 to May 2017. The data in this retrospective descriptive study was collected from urine culture records of the Microbiology Department, CMH Multan. Only those urine specimens who revealed positive gram-negative bacilli cultures were included in the study. Drug susceptibility patterns of these isolates were recorded against routinely used antibiotics (e.g. Nitrofurantoin, Imipenem, Sulbactum-cefoperazone, Gentamicin and Ciprofloxacin) and evaluated accordingly. RESULTS: A total of 1703 urine specimens were submitted for culture and antibiotics susceptibility testing during the period of study. A total of 128 specimens showed growth of gram-negative rods. Imipenem (95% sensitivity), Sulbactam- Cefoperazone (88% sensitivity) and Nitrofurantoin (87% sensitivity) were highly effective antibiotics against the cultured gram-negative bacilli in the study. CONCLUSION: This study showed that E. coli is the commonest cause of urinary tract infection (UTIs), followed by Klebsiella and Enterobacter species among gram-negative bacilli in our set up. In-vitro efficacy of Imipenem, Sulbactam- Cefoperazone and Nitrofurantoin was found to be the highest against these gram-negative bacilli as compared to other antimicrobials. On the contrary, in-vitro efficacy of ciprofloxacin and gentamycin was found to be extremely low.

Dispersive liquid–liquid microextraction of 11-nor-Δ9-tetrahydrocannabinol-carboxylic acid applied to urine testing

Aim: THC-COOH is the major metabolite of Δ9-tetrahydrocannabinol commonly tested in urine to determine cannabis intake. In this study, a method based on dispersive liquid–liquid microextraction was developed for testing THC-COOH in urine. Materials & methods: Hydrolyzed urine specimens were extracted via dispersive liquid–liquid microextraction with acetonitrile (disperser solvent) and chloroform (extraction solvent). Derivatization was performed with N,O-Bis(trimethylsilyl)trifluoroacetamide with 1% trichloro(chloromethyl)silane. Analysis was performed by GC–MS/MS. Results: The method showed acceptable linearity (5–500 ng/ml), imprecision (<10.5%) and bias (<4.9%). Limits of detection and quantitation were 1 and 5 ng/ml, respectively. Twenty-four authentic samples were analyzed, with 22 samples being positive for THC-COOH. Conclusion: The proposed method is more environmentally friendly and provided good sensitivity, selectivity and reproducibility.

Spot Urine Uric Acid to Creatinine Ratio used in the Estimation of Hyperuricosuria in the Young Korean Population

Background: Uric acid levels in urine are measured using urine specimens 24 hours or by uric acid glomerular filtration rate (UAGFR) with spot urine, which additionally requires a blood sample. This study aimed to investigate whether urinary uric acid creatinine ratio (UUACr) obtained by spot urine alone could be recognized as a substitute for UAGFR value, and hyperuricosuria can be screened by UUACr. UUACr is known to vary with age and regional differences. This study focused on the reference value of each value in Korean young populations.Method: We enrolled Korean subjects 1–20 years with normal kidney function, from a single hospital, classified into 5 age groups, 1–5 years, 6–8 years, 9–12 years, 13–15 years, and 16–20 years. We checked spot urine uric acid, creatinine and serum uric acid, creatinine levels on the same day from February 2014 to December 2018. We measured the average of UAGFR and UUACr in each groups. The UUACr cut-off value of the upper 2 standard deviation (SD) of UAGFR were taken.Results: The upper 2 SD of UUACr (mg/mg) and UAGFR (mg/dL) were determined in all age groups. UUACr decreased with grown up (P=0.000), but UAGFR were not statistically different among the groups. UUACr and UAGFR were not significantly different by gender. UUACr and UAGFR were positively correlated; UUACr cut-off value of upper 2 SD UAGFR (0.54 mg/dL) was 0.65 mg/mg in total age.Conclusions: UUACr could potentially be used to screen for hyperuricosuria.

Frequency and antimicrobial resistance pattern of bacterial isolates from patients with COVID-19 in two hospitals of Zanjan

Background and Objectives: The outbreak of COVID-19 has been challenging the global health systems. As one of the major associated concerns, microbial co-infections and antimicrobial resistance play critical roles in the prognosis of the disease. This study aims to evaluate co-infections in COVID-19 patients regarding drug resistance. Materials and Methods: A total number of 5530 Real Time PCR-confirmed COVID-19 cases, who were admitted to two major educational Hospitals in Zanjan, Iran, from February 2019 to February 2020 were included. Respiratory, blood and urine specimens were collected and cultured on selective media. Subsequently, isolates identification, disc diffusion suscep- tibility tests, and data analysis were carried out. Results: Bacterial and fungal co-infections were confirmed in 423 patients (8.1%). Co-infections were more prevalent among females (53.2%) than males (46.8%). Coinfected patients had a significantly higher mortality rate compared to those without co-infections (54.8% vs. 12.2%, P<0.001). Acinetobacter baumannii was the most prevalent bacteria isolated from respiratory tract (15.4%) and blood (2.1%). Escherichia coli (12.5%) was the most frequent bacteria in urine. Fungal co-in- fection was confirmed in 174 (3.36%) patients. Gram-negative bacteria were highly sensitive to colistin (97.85%) and wide- ly resistant to cefixime (91.79%) and trimethoprim-sulfamethoxazole (89.64%). Gram-positive bacteria were considerably sensitive to vancomycin (68%) and nitrofurantoin (66%). Tetracycline and ampicillin were the least effective antibiotics for Gram-positive bacteria with respective resistance rates of 90.91% and 83.33%. Conclusion: Given the high incidence of bacterial co-infections in COVID-19 patients, it is important to develop rapid and efficient diagnostic, therapeutic and disinfection guidelines to control these infections in the hospitals.

Bacterial uropathogens and burden of antimicrobial resistance pattern in urine specimens referred to Ethiopian Public Health Institute

Background Urinary tract infections (UTIs) are the leading causes of morbidity in the general population, and is the second most common infectious disease after respiratory infections. Appropriate antibiotic therapy is essential to achieving good therapeutic results. Therefore, the purpose of this study was to investigate the profile of pathogens cultured from urinary tract infections and to determine their resistance profiles to commonly prescribed antibiotics. Method A cross-sectional study was carried out at the National Referral Laboratory of the Ethiopian Institute of Public Health from January 2017 to December 2018. All positive cultures were characterized by colony morphology, Gram stain, and standard biochemical tests. The antimicrobial susceptibility test of the isolate was performed using the Kirby- Bauer disk diffusion test on Muller-Hinton agar. In addition, bacterial identification, antimicrobial susceptibility testing and phenotypic detection of MDR were performed with VITEK 2 Compact according to the manufacturer’s instructions. Result Out of 1012 cultured urine specimens, 325 (32.1%) was showed significant bacteriuria. The overall prevalence of UTIs was 325(32.1%) and the highest prevalence rate was obtained from 21–30 years age group 73(22.5%). Among UTIs patients, 583(57.6%) were females and 429(42.4%) were males. The UTIs of 179 (55%) women is relatively higher than that of men 146 (45%). Among 325 isolates, Gram-negative bacteria (GNB) appeared more frequently 252 (51.7%) than Gram-positive bacteria 63 (19.4%). In GNB, E. coli 168(66.7%), Klebsiella species 32(12.7%), and Enterobacter species 13 (5.2%) were dominated isolates whereas in GPB accounted for coagulase-negative staphylococcus (CoNS) 33(52.4%), Enterococcus species 16(25.4%), and Staphylococcus aureus 10(15.9%). Major of the isolates showed high levels of antibiotic resistance to commonly prescribed antimicrobials. Imipenem, Amikacin, and Nitrofurantoin were the most sensitive antibiotics for Gram-negative isolates while Nitrofurantoin, clindamycin, and Gentamycin were effective against gram-positive uropathogens. Overall, 156/256(60.9%), 56/256(22.4%), 10/256(4%) of gram-negative isolates were MDR, XDR, and PDR respectively while among the GPB isolates, 34/63(53.1%), 10/63(15.8%), and 1/63(1.6%) were MDR, XDR, and PDR isolates respectively. Among the tested bacterial strains, 190/319 (59.5%) were MDR, 66/319 (20.7%) strains were XDR, and 11/319 (3.45%) were PDR isolated. Conclusion The prevalence of urinary tract infection was high, and Gram-negative organisms were the most common causes of UTIs in this study. It was found that the resistance to commonly used antibiotics is very high. Early detection and close monitoring of MDR, XDR, or even PDR bacterial strains must be started by all clinical microbiology laboratories to reduce the menace of antimicrobial resistance that is now a global problem.

Post-exercise urinary alpha-1 acid glycoprotein is not dependent on hypoxia

Introduction: Proteinuria is a transient physiologic phenomenon that occurs with a range of physical activities and during ascent to altitude. Exercise intensity appears to dictate the magnitude of post-exercise proteinuria; however, evidence also indicates possible contributions from exercise-induced hypoxemia or reoxygenation. Utilizing an environmental hypoxic chamber, this crossover designed study aimed to evaluate urinary alpha-1 acid glycoprotein (α1-AGP) excretion pre/post exercise performed in hypoxia and normoxia. Methods: Sixteen individuals underwent experimental sessions in normoxia (NOR, 20.9% O2) and hypoxia (HYP, 12.0% O2). Sessions began with a 2-hour priming period before completing a graded maximal exercise test (GXT) on a cycle ergometer, which was followed by continuation of exposure for an additional 2 hours. Physiologic responses (i.e., blood pressure, heart rate, and peripheral oxygenation), Lake Louise Scores, and urine specimens (analyzed for albumin and α1-AGP) were collected pre- and post-exercise (after 30, 60, and 120 minutes). Results: Peak power output was significantly reduced in HYP (193 ± 45 W) compared to NOR (249 ± 59 W, p < 0.01). Post-exercise urinary α1-AGP was greater in NOR (20.04 ± 14.84 μg•min-1) compared to HYP (15.08 ± 13.46 μg•min-1), albeit the difference was not significant (p > 0.05). Changes in urinary α1-AGP from pre- to post-30 minutes were not related to physiologic responses or performance outcomes observed during GXT in NOR or HYP. Conclusion: Despite profound systemic hypoxemia with maximal exercise in hypoxia, post-exercise α1-AGP excretion was not elevated above levels observed following normoxic exercise.

67. Transitioning from Permissive to Restrictive Urine Reflex Criteria: Compiling the Data

Background Reflex urine cultures (UCx) are a diagnostic stewardship practice that limit the progression of UCx to specimens that meet pre-defined urinalysis criteria, but there is no widely recommended threshold for culture. At our institution, urinalyses (UAs) are reflexed to UCx for positive nitrites, leukocyte esterase, presence of bacteria, or ≥5 white blood cells per high powered field (WBC/hpf). Our aim is to assess if a more restrictive criteria of &gt;10 WBC/hpf would result in missed UTI diagnoses. Methods We performed a retrospective chart review of a systematic sampling of urine specimens collected from July 2018 to June 2019 in the emergency department and adult inpatient units. Inclusion criteria were UA with a WBC/hpf of 5-10 – samples that would not reflex to culture under our proposed criteria – and a UCx. We recorded signs, symptoms and antibiotic use via chart review. Positive UCxs were defined as ≥10e5 CFU/mL of bacterial growth (BG) and these cases were assessed using standardized CDC UTI definitions. Results 486 urine specimens with &lt; 10e5 CFU/mL BG and 96 with ≥10e5 CFU/mL BG met inclusion criteria. Chart review was performed on 99 cases. 81 (82%) specimens had negative UCxs and 18 (18%) were positive. 45% had documented localizing UTI symptoms. 26% of all urine studies were sent for an indication of fever, 15% for altered mental status (AMS), and 8% for malaise. Among the 18 patients with positive UCxs, 11 (61%) met UTI criteria. Among the 81 patients with negative UCxs, 33/81 (41%) had a local symptom compatible with UTI. 7/81 (9%) patients had positive tests from other body sites; all 7 of these UCxs were sent for a new or worsening fever. Conclusion Of the 99 UCxs reviewed, less than half had a urinary symptom consistent with UTI, and almost half of studies were sent for non-specific indications such as fever, which suggests reflex UCxs are overutilized at our institution. However, our data demonstrate that a more restrictive UCx criteria may not be the solution, as at least 11 clinically significant UTIs would have been missed under the new criteria. We recommend improved clinical decision support tools and more data to validate restrictive reflex UCx criteria before their implementation. Disclosures All Authors: No reported disclosures

Mapping of population disparities in the cholangiocarcinoma urinary metabolome

Phenotypic diversity in urinary metabolomes of different geographical populations has been recognized recently. In this study, urinary metabolic signatures from Western (United Kingdom) and South-East Asian (Thai) cholangiocarcinoma patients were characterized to understand spectral variability due to host carcinogenic processes and/or exogenous differences (nutritional, environmental and pharmaceutical). Urinary liquid chromatography mass spectroscopy (LC–MS) spectral profiles from Thai (healthy = 20 and cholangiocarcinoma = 14) and UK cohorts (healthy = 22 and cholangiocarcinoma = 10) were obtained and modelled using chemometric data analysis. Healthy metabolome disparities between the two distinct populations were primarily related to differences in dietary practices and body composition. Metabolites excreted due to drug treatment were dominant in urine specimens from cholangiocarcinoma patients, particularly in Western individuals. Urine from participants with sporadic (UK) cholangiocarcinoma contained greater levels of a nucleotide metabolite (uridine/pseudouridine). Higher relative concentrations of 7-methylguanine were observed in urine specimens from Thai cholangiocarcinoma patients. The urinary excretion of hippurate and methyladenine (gut microbial-host co-metabolites) showed a similar pattern of lower levels in patients with malignant biliary tumours from both countries. Intrinsic (body weight and body composition) and extrinsic (xenobiotic metabolism) factors were the main causes of disparities between the two populations. Regardless of the underlying aetiology, biological perturbations associated with cholangiocarcinoma urine metabolome signatures appeared to be influenced by gut microbial community metabolism. Dysregulation in nucleotide metabolism was associated with sporadic cholangiocarcinoma, possibly indicating differences in mitochondrial energy production pathways between cholangiocarcinoma tumour subtypes. Mapping population-specific metabolic disparities may aid in interpretation of disease processes and identification of candidate biomarkers.

Biofilm formation by Candida albicans and Candida glabrata isolated from urine specimens of diabetic Iraqi women

Biofilms are multicellular communities where microorganisms are grown and form an extracellular matrix that protects the pathogenic microorganisms from the immunity system and antimicrobial agents. This study is aimed to identify Candida spp. isolated from urine specimens by using traditional techniques, germ tube, growth on corn meal agar medium and chromogenic medium then determine the ability of isolates to producing biofilm by tube method (TM) and congo red agar method (CRA). In our study urine specimens were obtained from 174 diabetic females in the period of six months at the Al-Wafa Specialized Center for Diabetes and Endocrinology, Mosul city, Iraq. Out of the total 174 specimens, yeast species were isolated from 56 (32.2 %) specimens. Out of the 56 isolates, 50 isolates were positive for Candida spp., especially C. glabrata which appeared maximum in 30 isolates (60 %) and followed by C. albicans 18 isolates (36%) and C. krusei 2 isolates (4 %). In the TM method for biofilm detection, C. albicans showed 16 isolates (88.9%) positive for biofilm formation followed by 29 isolates (96.7%) of C. glabrata. Furthermore, in the CRA method, all isolates (100%) of C. albicans were negative followed by 27 isolates (90%) of C. glabrata, whereas only 3 isolates (10%) of C. glabrata were positive. We can conclude that TM is the best conventional method and is sensitive to detect biofilm-forming yeast when compared with the CRA method.