Cystine Lactose Electrolyte Deficient Research Articles

A-295 New Transport Media to Preserve Viable Parasites, Bacteria and Fungus

Abstract Background Patients with various infections not always show positive results in their cultures. Therefore, the purpose of this study is to show that a transport media is needed to preserve bacteria, parasites and fungi, which can help patients when collecting samples to determine their infections, the most common urine or feces. The microorganisms can be kept immediately in the transport media when they are taken by the patient and the first step is very important to keep them alive. The preanalytical step is essential for a good diagnostic and accurately determines their etiology. Methods The transport media that I am referring to, was designed by our laboratory. It is a tube with an enriched broth, it can be kept at room temperature for a year. To show that this transport media can be trusted, different samples were tested such as: Urine, semen, wounds, eye secretions and feces were tested at different temperatures: refrigerator 2°C, environment 18°C and in a “kangaroo” corporal temperature or an incubator at 37°C. Also, times were tested between the collection and transport to the laboratory to be proceeded. To have a good diagnostic the sample should be smeared onto the transport media and kept there at least two or more hours at normal corporal temperature. There is no problem to send the tube at room temperature to the laboratory. All samples were smeared for bacterias in MacConkey agar, EMB (Eosin Methylene Blue) agar, CLED (Cystine-Lactose-Electrolyte Deficient) agar with Andrade Indicator, Blood agar, Chocolate agar, XLD agar. For fungi in Potato or Sabouraud agar. The parasites were studied in saline solution and parasitological lugol. Results The samples most studied were 1600 samples of feces, 700 urines and 100 other samples, such as genital, eye secretions, wounds. The percentage of positive etiology with this transport media was 67.9% of fungi versus 73.5% without the transport media. The aerobic bacteria and helminths eggs or adults can be in this transport media for many days, there is no difference and Giardia lamblia trophozoites can be kept as long as seven hours. Conclusions Nowadays, there are various commercial transport media for each microorganism. The great advantage of our transport media is used for three etiologies: Bacteria, parasites and fungi, plus can be kept at room temperature for several days. It is very easy to used, the tube of the transport media can be placed in a “kangaroo” style to keep the samples at corporal temperature. It is helpful in case the patient lives far away or takes the sample at night. The transport media can keep the samples at 37°C at all time at corporal temperature. This new transport media is essential to obtain accurate positive cultures and can give a better diagnostic, since the microorganisms are alive. Knowing the etiology, the treatment is accurate and the patient´s health is always necessary.

Detection and Antimicrobial Susceptibility of Candida Species Isolated from the Urine of Patients in a Tertiary Health Facility, Southwest Nigeria

Aims: This study aimed to identify and determine the antimicrobial susceptibility profiles of Candida species causing UTI among patients attending a tertiary Teaching Hospital, in southwest Nigeria. Study Design: Comparative cross-sectional study. Place and Duration of Study: Lagos State University Teaching Hospital, Ikeja, Nigeria between June 2017 and February, 2018. Methodology: A total of 250 participants whose provisional diagnosis was candiduria were recruited for this study. Urine samples were collected from consenting participants early in the morning into sterile wide mouth universal containers. These samples were cultured aerobically on Blood agar, Cystine-Lactose-Electrolyte Deficient (CLED) agar and Sabouraud dextrose agar (SAB) at 370C within 1 to 2 days. The isolates were profiled into species level using microscopic, biochemical test, chromogenic media (Chrom agar Candida) and Analytical Profile Index (API) 32C examination analysis. Results: An overall rate for Candidiasis in this study was 12.8% (32/250). The rate was higher in female 17.5% compared to 6.5% in their male counterparts (p=0.014). Highest rate of infections peaked at 28.6% among age group 83-92 years and lowest (6.7%) in age group 23-32 years. A total of 11 (4.4%) urine culture from participants produced C. albicans pure fungi isolates. However, by gender, this was statistically significant (p=0.003). A total of 81 bacterial (32.4%) and fungal isolates 32 (12.8%) were isolated and profiled. Distribution of Candida species indicated highest incidence in age range 31-40 years, followed by 21-30years and age 61-70 years age brackets. The isolated species were Candida albicans 11(34.4%), C. tropicalis 8(25%), C. parapsilopsis 6(19%), C. krusei 2(6.3%) and C. hellenica 1(3.1%). Sixty to seventy percent of fungal isolates were susceptible to ketoconazole and fluconazole while the susceptibility pattern of Candida species to itraconazole, terbinafine and nystatin varied between moderately susceptible to resistance. All the Candida isolates were resistant to griseofulvin. However, C. albicans was found to be the major Candida species causing Candidal urinary tract infection. The only C. hellenica isolated was resistant to all the antifungal drugs were approved except nystatin. Conclusion: High profile of Candida isolates and related UTI microbes were found in this study. There is need for advocacy for the use of API and Chrom agar Candida for routine diagnosis for Candida identification. Nevertheless, the Candida isolates were mostly susceptible to ketoconazole and Fluconazole but only the isolates were griseofulvin resistant.

Pediatric Uropathogens and their Antimicrobial Susceptibility Pattern: Experience from an Impoverished District of Karachi, Pakistan.

Urinary tract infection (UTI) is the most common infection of the pediatric age group. Several factors linked to higher prevalence include poor personal hygiene, improper sanitation, lower socioeconomic status, and malnourishment. In addition to having a worse quality of life, the 1.8 million children who live in Karachi's Korangi district are routinely exposed to such factors. The study aims to evaluate the frequency of UTI and distribution of uropathogens along with their antimicrobial susceptibility pattern in patients presenting to a pediatric tertiary care center in the Korangi district of Karachi, Pakistan. The study employed an observational cross-sectional design. The study was conducted at the Microbiology laboratory of Sindh Institute of Child Health and Neonatology, Karachi, Pakistan from 1st January to 15th August 2023. Urine samples of patients 1 to 16 years of age were collected via midstream clean catch method and of patients from birth up to 1 year were collected in urine collection bags. The samples were cultured on Cystine Lactose Electrolyte Deficient (CLED) agar and antibiotic susceptibility testing was performed using the Kirby-Bauer Disc Diffusion method. A total of 457 urine samples were collected, of which 90 (19.7%) were positive for significant uropathogens. With a mean age of 4.6 years, majority of the culture-positive patients were female (n = 72; 80%). Enterobacterales were the most frequently isolated (n = 88; 95.6%), of which Escherichia coli was the most common (73.9%; n = 68). Citrobacter (n = 7; 7.6%), Klebsiella (n = 6; 6.5%), Serratia (n = 4; 4.3%), Proteus (n = 2; 2.2%), Salmonella (n = 2; 2.2%), and Enterobacter (n = 1; 1.1%) were among the other Enterobacterales isolated. Meropenem and imipenem were the most effective in isolates from Enterobacterales (n = 88) followed by amikacin (n = 84), ciprofloxacin (n = 75), and piperacillin-tazobactam (n = 70). Ceftriaxone and cefixime exhibited moderate susceptibility (n = 69 and 52) whereas, amoxicillin-clavulanate was the least susceptible (n = 3). We report high frequency of UTI in our pediatric population with uropathogens and associated antimicrobial susceptibility pattern confirming to the existing trends of pediatric UTIs in Pakistan. In addition to valuable insights for treating patients under similar conditions, our study serves as a catalyst for further multi-center research in this area.

Prevalence and antimicrobial resistance of uropathogens in Karachi, Pakistan

Urinary tract infections (UTIs) are one of the major healthcare concerns causing an alarmingly high medical and financial burden in both developing and developed countries. There is a significant rise in multidrug-resistant (MDR) uropathogens, both in hospitalized and community settings, which threatens safe and effective therapy of these infections. The increased rates of resistance in UTIs requires the continuous surveillance of uropathogens in the specific area to inform safe and effective therapy. The present, retrospective, cross-sectional, descriptive study was carried out using samples collected between 01/06/2019 and 31/05/2020 in Karachi, Pakistan, during which, n = 1500 urine samples were collected. The samples were processed on Cystine Lactose Electrolyte Deficient (CLED) agar, identification was carried out by using standard biochemical tests and API 20E/20NE strips. Antimicrobial susceptibility testing was performed using standard disk diffusion test protocol, as per Clinical and Laboratory Standards Institute (CLSI) guidelines. Overall, n = 1189 urine samples (79.27%) showed significant bacterial growth. The median age of affected patients was 56 years (range: 1-100) with n = 811 females (68.21%), with patients between 61-80 years (n = 384; 32.29%) as the most numerous age group. Regarding uropathogen distribution, the overwhelming majority were Gram-negative bacteria (n = 986; 82.93%), the most common causative agent being Escherichia coli (n = 648; 54.49%), followed by Klebsiella spp. (n = 206; 17.33%) and Enterococcus spp. (n = 118; 9.92%). Resistance rates were highest for the tested fluoroquinolones (>70% for most species), trimethoprim-sulfamethoxazole, broad-spectrum penicillins, and cephalosporins, while fosfomycin, carbapenems and colistin largely retained their efficacy. The mitigation of UTIs and the emergence of resistance may be impeded by taking appropriate measures for the better management of patients; these interventions include improvements in the treatment recommendations, provision of health education, and continuous antimicrobial surveillance.

COMPARISON OF SUSCEPTIBILITY BETWEEN CONVENTIONAL FIRST LINE ANTIBIOTIC CO-TRIMOXAZOLE AND NEWER ANTIBIOTICS IN RECURRENT UNCOMPLICATED URINARY TRACT INFECTIONS

Objective: The current study aims to evaluate the susceptibility pattern of relatively older antibiotic Co-trimoxazole and its comparison with Levofloxacin, Nitrofurantoin, and Fosfomycin. Material and Methods: This cross-sectional study was conducted in Mardan Medical Complex, Mardan, and the Postgraduate Medical Education Department of Khyber Girls Medical College, Peshawar from April 2022 to September 2022. Both male and female patients, above the age of 15 years with recurrent uncomplicated urinary tract infections were included in the study. The samples were inoculated onto CLED (Cystine-Lactose-Electrolyte-Deficient) Agar, a differential culture medium. The grown bacteria were identified, using Gram staining and BIOMÉRIEUX® API® 10S kits. Minimum inhibitory concentrations (MIC) were determined by the Agar dilution method; as per standard protocol. The results were compared among Co-trimoxazole, Levofloxacin, Nitrofurantoin, and Fosfomycin using statistical tests. Results: A total of 680 samples were received, of which 158 samples were culture-positive. The gender distribution of females and males was 63.3% and 36.7%, respectively. A predominant proportion of the patient cohort manifested within the age range of 21-40 years, with the subsequent highest representation observed in the 41-60 year age group. The isolated organisms were E. coli (74.1%), Klebsiella (10.8%), Pseudomonas (5.1%), Enterococci (6.3%), Proteus species (2.5%), and Citrobacter (1.3%). Based on MIC analysis, 77.2% of isolates were found to be sensitive to Co-trimoxazole, 52.5% to Levofloxacin, 86.7% to Nitrofurantoin, and 90.5% to Fosfomycin. When comparing antibiotics, Co-trimoxazole displayed significantly higher effectiveness against the isolates compared to Levofloxacin (p-value 0.004). However, in comparison to Nitrofurantoin and Fosfomycin, Co-trimoxazole exhibited lower effectiveness, with respective p-values of 0.000 and 0.007. Conclusion: The study revealed that the susceptibility of bacterial isolates to Co-trimoxazole is significantly higher than that to Levofloxacin but lower than that to Nitrofurantoin and Fosfomycin. KEYWORDS Urinary Tract Infections, Drug Resistance, Culture And Sensitivity, Minimum Inhibitory Concentration

The Antimicrobial Resistance (AMR) Rates of Enterobacterales in a Rural Hospital from the Eastern Region, Ghana: A Retrospective Study, 2022.

Low- and middle-income countries bear a disproportionate burden of antimicrobial resistance and often lack adequate surveillance due to a paucity of microbiological studies. In this 2022 study, our goal was to contribute to a more precise antimicrobial treatment by understanding the prevalence of resistance in a rural environment, promoting antibiotic stewardship, and raising awareness about antimicrobial resistance. We assessed the prevalence of Multidrug-Resistant (MDR) and Extensively Drug-Resistant (XDR) Enterobacterales in clinical samples from 2905 patients being treated at Saint Dominic's Hospital, Akwatia, in the countryside of the Eastern Region, Ghana, in the year 2022. To this purpose, the samples were cultured on agar plates prepared in the laboratory using purified Oxoid™ Thermo Scientific™ agar (Thermo Fisher Scientific; Waltham, MA, USA). Cystine Lactose Electrolyte-Deficient (CLED) agar was used for urine samples, while blood agar, chocolate agar, and MacConkey agar were used for the rest of the specimens tested (HVS, blood, BFA, sputum). Antimicrobial susceptibility was determined on site using the disc diffusion method (Kirby-Bauer test). MDR bacteria accounted for more than half (53.7%) of all microorganisms tested for three or more antibiotics and 37.3% of these were XDR. Multivariate regression analysis was performed to identify risk factors associated with acquiring MDR/XDR bacteria. The results showed an increased likelihood of MDR acquisition linked to being male (OR 2.39, p < 0.001 for MDR and OR 1.95, p = 0.027 for XDR), higher age (OR 1.01, p = 0.049 for MDR), non-sputum samples (OR 0.32, p = 0.009 for MDR), and urine samples (OR 7.46, p < 0.001 for XDR). These findings emphasize the urgency for surveillance and control of antimicrobial resistance; to this end, making accurate diagnostics, studying the microorganism in question, and conducting susceptibility testing is of the utmost importance.

Bacterial etiology of urinary tract infections in patients treated at Kenyan health facilities and their resistance towards commonly used antibiotics.

Evidence-based empirical antibiotic prescribing requires knowledge of local antimicrobial resistance patterns. The spectrum of pathogens and their susceptibility strongly influences guidelines for empirical therapies for urinary tract infections (UTI) management. This study aimed to determine the prevalence of UTI causative bacteria and their corresponding antibiotic resistance profiles in three counties of Kenya. Such data could be used to determine the optimal empirical therapy. In this cross-sectional study, urine samples were collected from patients who presented with symptoms suggestive of UTI in the following healthcare facilities; Kenyatta National Hospital, Kiambu Hospital, Mbagathi, Makueni, Nanyuki, Centre for Microbiology Research, and Mukuru Health Centres. Urine cultures were done on Cystine Lactose Electrolyte Deficient (CLED) to isolate UTI bacterial etiologies, while antibiotic sensitivity testing was done using the Kirby-Bauer disk diffusion using CLSI guidelines and interpretive criteria. A total of 1,027(54%) uropathogens were isolated from the urine samples of 1898 participants. Staphylococcus spp. and Escherichia coli were the main uropathogens at 37.6% and 30.9%, respectively. The percentage resistance to commonly used drugs for the treatment of UTI were as follows: trimethoprim (64%), sulfamethoxazole (57%), nalidixic acid(57%), ciprofloxacin (27%), amoxicillin-clavulanic acid (5%), and nitrofurantoin (9%) and cefixime (9%). Resistance rates to broad-spectrum antimicrobials, such as ceftazidime, gentamicin, and ceftriaxone, were 15%, 14%, and 11%, respectively. Additionally, the proportion of Multidrug-resistant (MDR) bacteria was 66%. High resistance rates toward fluoroquinolones, sulfamethoxazole, and trimethoprim were reported. These antibiotics are commonly used drugs as they are inexpensive and readily available. Based on these findings, more robust standardised surveillance is needed to confirm the patterns observed while recognising the potential impact of sampling biases on observed resistance rates.

Evaluation of chromogenic agar medium, can it be a suitable alternative to conventional culture system for identification of uropathogens?

Urinary tract infections (UTI) account for major proportion of outpatient load and hospital admission globally. In most of the clinical microbiology laboratories MacConkey agar (MAC) and Cystine lactose electrolyte-deficient (CLED) agar are being used for identification of uropathogens. The main objective of the present study was to evaluate the usefulness of HiCromeTM UTI by comparing isolation rate and presumptive identification of uropathogens against CLED and MAC agar. This study was conducted over a period of three months on 672 non-duplicate midstream and/or catheter-catch urine samples. All samples were inoculated on to HiCromeTM UTI, CLED agar and MacConkey agar. Among the 672 samples received for culture, 113 (16.8%) showed significant growth. Among the 672 samples, 95 (14.1%) showed growth of a single organism while 18 (2.7%) showed polymicrobial growth. The rate of isolation and presumptive identification of the isolates and polymicrobial growth was found significantly higher on HiCromeTM UTI Agar. HiCromeTM UTI Agar has the potential to streamline processing of samples for urine culture in a way that will reduce the workload for technicians, reduce turnaround time which in turn will benefit the laboratory ultimately leading to better patient care.

Biofilm formation by ESBL-producing uropathogenic E. coli

Introduction: E. coli is considered to be the leading cause of urinary tract infections. (UTIs) and is responsible for both community acquired and hospital acquired UTIs. The ability of this bacterium to form biofilm on various devices like catheters plays an important role in progression of the disease. Biofilm production in extended spectrum beta lactamases (ESBLs) producing organisms provides a favourable environment for the exchange of resistance genes and may also facilitate widespread dissemination.&#x0D; Methods: The urine samples were collected from the suspected cases of urinary tract infections and were cultured on cystine lactose electrolyte deficient (CLED) agar plates to isolate the desired UPEC strains. All the isolated E. coli strains were subjected to ESBL detection by double disk approximation method and to biofilm formation by tissue culture plate method of biofilm detection. All the results were statistically analysed using k2 test with p value at &lt;0.05 level of significance.&#x0D; Results: Out of all the gram negative bacilli isolated from urine samples, E. coli alone accounted for 90% of the isolates. Of all the UPEC isolates subjected to ESBL detection 50% of the strains turned out to be the ESBL producers and 35% of the strains showed biofilm formation.&#x0D; Conclusion:&#x0D; &#x0D; The majority of the work in literature sites the data samples collected from indoor patients, where these isolates coli strains which produced ESBLs were mainly a strong biofilm former as compared to non-ESBL producer strains.&#x0D; In this study in spite of all the coli strains being outpatient isolates still the correlation between ESBL production and biofilm formation holds true.&#x0D; We found a significant correlation between biofilm formation and ESBL production. The detection of ESBL production and biofilm formation pattern will help in development of newer and more effective treatment.&#x0D;

Symptomatic Pathogen Frequency and Antibiogram Patterns of Bacterial Isolates in Urinary Tract Infections, Sirajganj Sadar, Bangladesh

Urinary tract infections (UTIs) are broadly common in inpatient and outpatient males and females of different ages and comprise 40% of the nosocomial infections that collectively cause 150 million deaths per year. The right care is obligatory if the patients do not want to die, but this is hard to do because the things that make people sick are getting better at resisting antibiotics (AMR). Collected specimens were examined microscopically and cultured on Cystine Lactose Electrolyte Deficient (CLED) agar to isolate the pathogens. Isolated pathogens were identified through colony morphology, microscopic studies, and biochemical indications. Lastly, antimicrobial responsiveness patterns of the pathogens were determined by the disc diffusion method to find possible antibiotics that could treat the disease well. Among 17.67% of positive cultures, 73.58% were females, and 26.42% were males, dominated by the age group ≥37 years (49.06%). E. coli (56.6%), Enterococcus faecalis (26.4%), Pseudomonas aeruginosa (7.5%), Staphylococcus aureus (3.8%), Acinetobacter baumannii (3.8%). Moreover, Serratia marcescens (1.9%) were the isolated pathogens, with 69.8% Gram-negative and 30.2% Gram-positive. The most effective antibiotics were amikacin (88.68%), levofloxacin (88.68%), ciprofloxacin (86.79%), gentamicin (84.91%), and imipenem (84.91%). The least effective antibiotics were mecillinam (50.94%), cefuroxime (37.74%), ceftazidime (37.74%), meropenem (35.85%), and cefotaxime (33.96%). To treat and suggest antimicrobials, routine and emphatic research about urogenital pathogens and their antibiotic susceptibilities is required. Patients should be prescribed appropriate antimicrobial therapies after completing a standard test of pathogen identification and antibiotic resistance pattern determination. This study would be of extensive importance to patients and physicians in picking appropriate antimicrobial therapies for empiric treatment.

Significant bacteriuria among requested repeat urine samples and its clinical correlation

Background and Objectives:Urinary tract infections (UTI) are the most common bacterial infections in both outpatient and inpatient department received for routine bacterial culture and sensitivity. We looked for significant bacteriuria in requested repeat urine sample after primary urine culture yielded significant growth (>105 CFU/ml) of ≥3 types of colonies. Also studied, different isolates grown with their sensitivity pattern and contamination rates of urine samples from different departments.Materials and Methods:In routine, primary urine cultures yielding ≥3 types of colonies on Cystine Lactose Electrolyte Deficient (C.L.E.D) were requested for repeat samples, collected with aseptic precautions after proper instructions. Data was analyzed for the Microbiological profile and its clinical correlation.Results:Among 617 received requested urine samples, 292 (47.3%) yielded significant bacteriuria. Clinical details were available for 252 cases out of which 100 (39.7%) showed asymptomatic bacteriuria, 87 (34.5%) complicated UTI and 65 (25.7%) uncomplicated UTI. Null hypothesis was rejected as 292 (47.3%) of the received repeat samples showed significant bacteriuria and 325 (53%) showed normal flora/no growth i.e. there is a 50% chance of getting either a positive culture or normal flora/no growth in repeat urine samples after the primary urine culture showed ≥3 types of colonies. It indicates the importance of requesting repeat urine samples for an accurate urine culture report. Male patients were significantly associated with significant bacteriuria and complicated UTI (p= 0.001). Escherichia coli (n=112, 28%) was the most common followed by Klebsiella species (n=66, 16.4%) and Enterococcus species (n=69, 17.2%). 183 (45.6%) isolates were Multi-Drug Resistant (MDR) Gram Negative Bacilli (GNBs), Escherichia coli (50.3%) being most common. Vancomycin Resistant Enterococcus (VRE) (n=8, 2.0%) was also isolated.Conclusion:Our study justifies the rationale for asking a repeat urine samples which helps in providing an appropriate microbiological report with antibiotic sensitivity pattern, hence preventing unwanted reporting of commensals/contaminants facilitating evidence based therapy.

The use of cinnamon oil as antibacterial agent to eliminate some antibiotic-resistant bacteria isolated from water sources

The use of cinnamon oil as antibacterial agent to eliminate some antibiotic-resistant bacteria isolated from water sources

Fosfomycin Sensitivity Pattern among Uropathogens Isolated from Patients Visiting Day Care Facility of Sushila Tiwari Hospital in Kumaun Region, Uttarakhand, India

Introduction: Urinary Tract Infections (UTI) are one of the most common infections responsible for antibiotic resistance. There are limited antibiotics options for treating the cases due to Multi Drug Resistant (MDR) bacteria. Fosfomycin is being used for treating UTIs and has shown promising results even against MDR pathogens. Aim: To determine the fosfomycin sensitivity pattern along with Minimum Inhibitory Concentration (MIC) against uropathogens by agar dilution method. Materials and Methods: The present cross-sectional study was conducted in the Department of Microbiology, Government Medical College Haldwani, Uttarakhand, India, between August 2017 to September 2019. Clean catch, mid stream urine samples were inoculated on Cystine Lactose Electrolyte Deficient (CLED) agar and incubated. The significant growths of pathogenic bacteria were subjected to antibiotic susceptibility testing. Fosfomycin (200 μg) disc was used in Kirby-Bauer disc diffusion testing. Fosfomycin trometamol MIC was determined by agar dilution method as per Clinical and Laboratory Standards Institute (CLSI) guidelines. The data collected in the study were analysed by using Statistical Package for the Social Science (SPSS) software version 20.0. Results: Significant growth of pathogenic bacteria was observed in 365 out of total 2725 urine samples. E.coli (72.32%) was the leading isolate followed by Enterococcusspecies (10.41%). Fosfomycin was recorded as the most active antibiotic against all the bacterial pathogen with 85-100% susceptibility except Proteus species (40%) in disc diffusion method. The MIC of fosfomycin was recorded between 4-64 against most of the isolates by agar dilution method. Conclusion: Fosfomycin is the most active antibiotic against all the uropathogens in the study setup and can be included in empirical treatment of day care patients along with nitrofurantoin.

Plasmid Profiling and Occurrence of β-Lactamase Enzymes in Multidrug-Resistant Uropathogenic Escherichia coli in Kathmandu, Nepal.

IntroductionExtended-spectrum β-lactamases (ESBL) among Gram-negative bacteria, predominantly Escherichia coli (E. coli), in Nepal, have been rising. The main objectives of this study were to determine the prevalence of uropathogenic E. coli, antibiotic resistance, ESBLs, ABLs (AmpC type β-lactamases), MBLs (metallo-β-lactamases) and KPCs (Klebsiella pneumoniae carbapenemases) and their correlation with plasmid profiling patterns among patients with urinary tract infections in a tertiary hospital in Kathmandu, Nepal.MethodsThe mid-stream urine samples collected from patients were inoculated in cystine–lactose–electrolyte-deficient (CLED) agar plates. E. coli producing ESBLs, ABLs, MBLs/KPC were identified phenotypically using standard microbiological methods. Plasmids were extracted by alkaline lysis method from E. coli isolates and profiled using agarose gel electrophoresis.ResultsOut of the total 2661 urine samples, E. coli were isolated in 64.34% (507/788), among which 170 (33.53%) were multidrug-resistant (MDR) isolates. All MDR isolates were resistant to amoxicillin and third-generation cephalosporins but were highly sensitive to imipenem (94.12%, 160/170), amikacin (92.94%, 158/170) and nitrofurantoin (86.47%, 147/170). Among 170 MDR isolates, 78.2% (133/170) were ESBLs, 46.3% (50/170) were AmpC, 11.2% (19/170) were MBL and 0.6% (1/170) were KPC producers. Coproduction of β-lactamases was detected in 24.12% (41/170) of isolates. E. coli isolates showed one plasmid (>33.5 kb), which was present in all the isolates. Overall, 44 different plasmid profile groups were identified based on molecular weight and number of plasmids. β-Lactamase producers were relatively resistant to the higher number of antibiotics tested (≤10) than non-producers (≤8), and the number of plasmids were higher in β-lactamase producers (≤7) than those in non-producers (≤5).ConclusionThe higher prevalence of the ESBLs, AmpCs, KPCs and MBLs along with their coproduction in E. coli isolates highlights the importance of routine surveillance of ESBLs, AmpCs, KPCs and MBLs in microbiology laboratories using various phenotypic methods.

Phenotypic detection of Extended Spectrum β-Lactamases (ESBL) among gram negative uropathogens reveals highly susceptibility to imipenem.

Objective:We aimed to determine antibiotic susceptibility patterns of ESBL- and non-ESBL bacteria isolated from pregnant women with UTI in antenatal wards in Khartoum State, Sudan.Methods:This cross-sectional study was conducted during April-July 2016 at different hospitals in Khartoum State. Mid-stream urine samples were obtained from 150 hospitalized pregnant women and cultured on CLED (Cystine Lactose Electrolyte Deficient) agar. Microorganisms were identified using standard microbiological procedures. Isolated Gram-negative bacteria were tested for antibiotic susceptibility and ESBL screening using modified Kirby- Bauer method and Double Disc Synergy Test (DDST) respectively.Results:Urine culture revealed positive results in 33/150 (22%) and the most prevalent isolates were Gram negative bacteria (18/33, 54.5%). Among gram-negative bacteria, isolates of E. coli were the most prevalent accounting 66.6% (12/18) followed by K. pneumoniae (4/18, 22.2%) and K. oxytoca (2/18, 11.1%). ESBL was detected in 8/18 (44.4%) of the Gram-negative isolates. Of note, imipenem was the most susceptible antibiotic for ESBL-producer and non-ESBL producer Gram negative isolates, accounting 100% susceptibility for both bacterial groups. Overall susceptibility rates were also high for ciprofloxacin (13/18, 72.2%). In other hand, co-trimoxazole and amoxicillin showed high resistance pattern for ESBL-producer and non-ESBL producer isolates; 27.8%, 44.4% and 38.9%, 38.9% susceptibility rates of co-trimoxazole and amoxicillin for ESBL-producer and non-ESBL producer isolates, respectively.Conclusions:Imipenem remains the most powerful option for ESBL- and non-ESBL bacteria causing UTIs in pregnant women. However, due to tremendous increase of antibiotic-resistant, antibiotic-susceptibility testing is recommended as a routine investigation for admitted pregnant women.

Yield of Urinary Pathogens and their Sensitivity Pattern at Paediatric Tertiary Care Facility, Karachi

Objective: To determine the yield of common pathogens isolated from urine specimens and their sensitivity pattern in children at National Institute of Child Health (NICH), Karachi.&#x0D; Methods: This was a retrospective descriptive study conducted in the department of paediatric neph- rology and pathology, National Institute of Child Health over a period of two years, from January 2014 to December 2015. All urine samples received in our laboratory from children admitted to nephrology and other departments during the study period were processed for culture and sensitivity. All samples were inoculated with cystine lactose electrolyte deficient (CLED) agar plates and biochemical charac- teristics of the specific pathogen were identified on biochemical tubes. The plates and tubes were in- cubated for 24 hours at 37ºC and growth of more than 105 CFU was considered as significant. These colonies were identified by morphology, lactose fermentation, Gram stain and biochemical tests. Anti- biotic sensitivity was performed by Kirby-Bauer method as per CLSI (Clinical and Laboratory Standards Institute) guidelines and sensitivity was recorded after 24 hours' incubation at 37ºC.&#x0D; Results: Out of 4107 samples processed, 1442 were positive giving a yield of 35% with 66% in girls and 34% in boys. E. coli was the most common pathogen (55%), followed by Pseudomonas (13.6%), Klebsiella (13,5%) and Candida albicans (11%). Majority of pathogens were sensitive to piperacillin- tazobactam (80-100%). Quinolones and aminoglycosides sensitivity was found in 65 to 85% and in 65 to 90% cases, respectively. Most of the pathogens (75%) were resistant to 3rd generation cepha- losporins except Citrobacter species.&#x0D; Conclusion: A high yield of common pathogens in urine samples and high resistance to common antibiotics was observed.

Correlation between biofilm formation and resistance toward different commonly used antibiotics along with extended spectrum beta lactamase production in uropathogenic Escherichia coli isolated from the patients suspected of urinary tract infections visiting Shree Birendra Hospital, Chhauni, Kathmandu, Nepal.

BackgroundEscherichia coli is the most predominant causative agent of urinary tract infection (UTI). Recently, increase in drug resistance among the uropathogenic bacteria has caused great problem in treatment of UTI. The main objective of this research is to determine the correlation between biofilm formation and resistance toward different commonly used antibiotics along with extended spectrum beta lactamase production in uropathogenic Escherichia coli.MethodsThe urine samples collected from the patients suspected of urinary tract infections (visiting Shree Birendra Hospital, Chhauni, Kathmandu, Nepal between July to December 2013) were cultured in cystine lactose electrolyte deficient (CLED) agar by using semi quantitative culture technique. Extended spectrum beta lactamase (ESBL) production was detected by combined disc diffusion technique and biofilm formation was detected by Congo red agar method. Chi-square test was applied and p-value < 0.05 was considered statistically significant.ResultsOut of 1480 urine samples, E. Coli was isolated from 208 (14.1 %) samples. Of total 69 (33.2 %) ESBL producing uropathogenic strains of E. coli, 20 (29 %) were strong biofilm producers, 22 (31.9 %) were moderate biofilm producers, 11 (15.9 %) were weak biofilm producers and 16 (23.2 %) were biofilm non producers. Whereas among 139 ESBL non producing E. coli, 22 (15.8 %) were strong biofilm producers, 20 (14.4 %) were moderate biofilm producers, 13 (9.4 %) were weak biofilm producers and 84 (60.4 %) were biofilm non producers. Among total 108 biofilm producing E. coli, maximum resistance was observed toward cephalexin followed by amoxicillin and highest susceptibility was seen toward amikacin.ConclusionThe ability of biofilm formation was found to be significantly higher in ESBL producing strains of E. coli than that in ESBL non producing strains (p < 0.05). There was higher resistance rate to antimicrobial agents among biofilm producing strains of E. coli than that in biofilm non producing strains. According to our antimicrobial susceptibility pattern for E. coli, to start preliminary treatment for UTI in Nepal, we recommend to use amikacin or nitrofurantoin. Further, for the treatment of the UTI, the antibiotics should be selected on the basis of the urine culture and sensitivity report.

Asymptomatic Bacteriuria Prevalence among Primary School Children in the Federal University of Technology, Akure (Futa), Ondo State, Nigeria

A study was carried out to determine the prevalence of bacteriuria among primary school children in The Federal University of Technology, Akure. One hundred and fifty (150) Mid-Stream Urine (MSU) samples were collected from both male and female pupils aged 5-11. Samples were examined macroscopically for colour and haematuria prior to the microbiological analysis on Cystine Lactose Electrolyte Deficient (CLED) agar using standard methods. Macroscopic examination showed 50.67% of urine samples with amber and clear; 14.67% with amber and cloudy, 6% with straw and cloudy and 24% with straw and clear. 2.67% of samples showed haematuria. Bacteria isolates encountered are Escherichia coli , Staphylococcus aureus , Klebsiella spp., and Pseudomonas aeruginosa . The predominant bacterial isolate was E. coli (56.8%), followed by S. aureus (18.9%), Klebsiella spp. (16.2%) and P. aeruginosa (8.1%). Overall prevalence of bacteriuria was 24.7%. Bacteriuria was higher among females (29.9%) than in males (11%). School children within the age group 10-11 yrs had the highest prevalence (20.0%), and in males; while age group 5-7 yrs had the highest asymptomatic bacteriuria prevalence (35.3%) and in females. This paper discussed the possible health implications of bacteriuria prevalence observed, and also emphasized the need for routine laboratory checks among children in the primary school.

Salmonella Enteric Fevers and Microbial Contamination in Public Eating Places in Makurdi, Nigeria.

Food, drinking water, table and utensil swab and hand washing water samples were examined for Salmonella sp contamination. Microbiological examination of samples using lactose broth, Cystine- Lactose-Electrolyte-Deficient (CLED) agar and Eosin-Methylene-Blue (EMB) agar was done to identify Salmonella sp, other organisms and indicator organisms. Indicator organisms identified were Aerobocter aerogenes and Escherichia coli. Other organisms include Streptococcus faecalis, Proteus sp and Staphylococcus aureus. 74.7% of examined samples showed Salmonella sp contamination while 89.8% of examined samples were contaminated with indicator organisms. Observed Salmonella sp were also examined biochemically to identify different species/strains. Species/strains identified were Salmonella typhi, S. paratyphi A, S. paratyphi B, S. cholerae-suis, S. enteritidis. There was a significant association between sanitary conditions and the occurrence of S. typhi in food, water, surroundings. It is very likely that poor hygiene and sanitary conditions encourage the occurrence of S. typhi in water, food, surroundings. Personal and environmental hygiene should be strictly adhered to especially in the study area so as to prevent and control S. typhi infections.

English

This research effort seeks to use doses of thioridaxine to enhance antibiotic sensitivity in a multidrug resistant (MDR) Staphylococcus aureus strain. Five axenic (pure) strains of S. aureus coded SA 1 to SA 5 were obtained from five infected midstream urine samples, inoculated on sterile cystine lactose electrolyte deficient (CLED) agar and stocked on sterile nutrient agar slants at 4°C in a refrigerator. Bacteria strains were sub-cultured on fresh sterile CLED agar and mannitol salt agar plates to confirm S. aureus strains. Gram staining, catalase test and coagulase test were done on the resulting colonies to further confirm the strains as S. aureus . Antibiotic susceptibility test was done by agar disc diffusion method using sterile Mueller- Hinton agar plates before and after treatment with laboratory dilutions of thioridaxine. S. aureus strains 1, 3 and 5 were multidrug resistant as they resisted 3 (37.5%), 3 (37.5%) and 4 (50.0%) of the antibiotics used. The highest (11.8±1.4 mm) and least (0.8±10.0 mm) zones of inhibition by all five strains were recorded for streptomycin and augmentin, respectively whereas, all five uropathogen strains resisted cloxacillin, they were sensitive to gentamycin, cotrimoxazole, chloramphenicol and streptomycin. After treatment with 2000 to 2240 ug/ml laboratory dilutions of thioridaxine, ≤50.0% loss of resistance was recorded for each of all seven dilutions but only 2240 ug/ml dilution recorded mean±S.E. loss of 56.2±17.8% for gentamycin, cotrimoxazole and streptomycin after treatment of SA 5 uropathogen. This was followed by resistance losses of 41.4±10.8 and 42.7±8.3% induced by 2080 and 2200 ug/ml dilutions, respectively. Cumulative effect of all dilutions resulted in 40.0±8.2 and 40.5±17.1% borderline resistance losses to cotrimoxazole and chloramphenicol, respectively. Minimum inhibitory concentration of chloramphenicol was lowered by 2080, 2160 and 2240 ug/ml dilutions of thioridazine by four-fold (7.5 ug), four-fold (7.5 ug) and two-fold (15 ug), respectively. Upon this, the medical/chemotherapeutic implications of these findings are discussed. Key words : In vitro, dilutions, thioridaxine, enhance, antibiotic, sensitivity, multidrug resistant Staphylococcus aureus.