scholarly journals Very rapid flow cytometric assessment of antimicrobial susceptibility during the apparent lag phase of microbial (re)growth

2018 ◽  
Author(s):  
Srijan Jindal ◽  
Harish Thampy ◽  
Philip J. Day ◽  
Douglas B. Kell

AbstractCells ofE. coliwere grown in LB medium, taken from a stationary phase of 2-4h, and reinoculated into fresh media at a concentration (105.mL-1or lower) characteristic of bacteriuria. Flow cytometry was used to assess how quickly we could detect changes in cell size, number, membrane energisation (using a carbocyanine dye) and DNA distribution. It turned out that while the lag phase observable macroscopically via bulk OD measurements could be as long as 4h, the true lag phase could be less than 15-20 min, and was accompanied by many observable biochemical changes. Antibiotics to which the cells were sensitive affected these changes within 20 min of reinoculation, providing the possibility of a very rapid antibiotic susceptibility test, on a timescale compatible with a visit to a GP clinic. The strategy was applied successfully to genuine potential Urinary Tract Infection (UTI) samples taken from a doctor’s surgery. The methods developed could prove of considerable value in ensuring the correct prescription and thereby lowering the spread of antimicrobial resistance.

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S537-S537
Author(s):  
Hyun Joo Jung ◽  
Shin Young Lee

Abstract Background Childhood urinary tract infection (UTI) may cause increased major morbidity and long-term clinical consequences. Extended-spectrum β-lactamase (ESBL) is produced by the members of the Enterobacteriaceae family, which are the primary infectious agents that cause UTI in children. Isolation of ESBL-producing Enterobacteriaceae (ESBL-E) typically occurred in healthcare facilities; however, the incidence of community-associated (CA) UTIs due to ESBL-E has increased worldwide. It has led to an increase in the use of carbapenems. In this study, we determine the characteristics of community-onset UTIs caused by ESBL-E in children to suggest non-carbapenem options for the treatment of childhood UTIs due to ESBL-E in order to preserve carbapenems. Methods A total of 2,157 isolates of ESBL-E were collected from children below 18 years old who were clinically certified UTI or urosepsis between January 2008 and August 2018 at tertiary university hospital in Korea. Their electronic medical records were retrospectively reviewed. Long-term healthcare facility stay within the preceding month and isolates recovered more than 72 hours after hospitalization were the criteria of healthcare-associated (HA) infection. Results The most common isolates were E. coli 1815 (84.2%) followed by K. pneumoniae 342 (15.8%). CA infection was detected in 1,513 of the 2157 ESBL-E (70.1%). The prevalence of CA ESBL-E infection increased significantly from 68 cases in 2008 to 325 cased in 2017. Antibiotic susceptibility test showed highest sensitivity to ertapenem, meropenem, and amikacin (>90%) followed by cefoxitin (82%), and piperacillin–tazobactam (TZP) (80.5%). CA E. coli showed higher sensitivity to amikacin and TZP compared with HA E coli. CA K. pneumoniae showed much higher sensitivity to TZP compared with HA K. pneumoniae. Of total ESBL-E, the antimicrobial resistance rate to aminoglycoside such as amikacin and gentamicin showed full sensitivity during the study period; furthermore, a rate of resistance to TZP has been decreasing over the years. Conclusion Identifying antibiotic susceptibility patterns of ESBL-E is a useful guide for treatment strategy of UTI. This study showed that there are non-carbapenem options for the treatment of CA ESBL UTI in children. Disclosures All authors: No reported disclosures.


2004 ◽  
Vol 171 (4S) ◽  
pp. 22-23
Author(s):  
Shingo Minagawa ◽  
Chikara Ohyama ◽  
Shingo Hatakeyama ◽  
Kazunari Sato ◽  
Shigeru Sato ◽  
...  

2018 ◽  
Vol 1 (2) ◽  
pp. 40-57
Author(s):  
Abdulghani Alsamarai ◽  
Shler Khorshed ◽  
Imad Weli

Background: Antibiotic resistance emerged as clinical problem challenge the effective treatment of infections. Virulence factor may play an important role in the influence of antimicrobial resistance. Objective: To determine the frequency of resistance gene in E. coli clinical isolates from women with urinary tract infection. Materials and Methods: Fifteen E.coli clinical isolates were tested by PCR to determine their molecular characterization. Results: The bla CTX –M gene was not detected in 6.7% out of the tested 15 E. coli clinical isolates from women with urinary tract infection. However, bla OXA gene was detected in all E. coli tested clinical isolates from pregnant women, female student and diabetic women with urinary tract infection. While bla TEM gene and bla SHV gene were not detected in 33.3% and 40% out of the tested E. coli clinical isolates respectively. Conclusions: Four types of ESBL genes were detected, and shows new trend of distribution, which indicated the predominance of OXA and CTX-M genes.


2018 ◽  
pp. 100-108
Author(s):  
Dinh Khanh Le ◽  
Dinh Dam Le ◽  
Khoa Hung Nguyen ◽  
Xuan My Nguyen ◽  
Minh Nhat Vo ◽  
...  

Objectives: To investigate clinical characteristics, bacterial characteristics, drug resistance status in patients with urinary tract infections treated at Department of Urology, Hue University Hospital. Materials and Method: The study was conducted in 474 patients with urological disease treated at Department of Urology, Hue Universiry Hospital from July 2017 to April 2018. Urine culture was done in the patients with urine > 25 Leu/ul who have symptoms of urinary tract disease or infection symptoms. Patients with positive urine cultures were analyzed for clinical and bacterial characteristics. Results: 187/474 (39.5%) patients had symptoms associated with urinary tract infections. 85/474 (17.9%) patients were diagnosed with urinary tract infection. The positive urine culture rate was 45.5%. Symptoms of UTI were varied, and no prominent symptoms. E. coli accounts for the highest proportion (46.67%), followed by, Staphycoccus aureus (10.67%), Pseudomonas aeruginsa (8,0%), Streptococcus faecali and Proteus (2.67%). ESBL - producing E. coli was 69.23%, ESBL producing Enterobacter spp was 33.33%. Gram-negative bacteria are susceptible to meropenem, imipenem, amikacin while gram positive are vancomycin-sensitive. Conclusions: Clinical manifestations of urinary tract infections varied and its typical symptoms are unclear. E.coli is a common bacterium (46.67%). Isolated bacteria have a high rate of resistance to some common antibiotics especially the third generation cephalosporins and quinolones. Most bacteria are resistant to multiple antibiotics at the same time. Gram (+) bacteria are susceptible to vancomycin, and gram (-) bacteria are susceptible to cefoxitin, amikacin, and carbapenem. Key words: urinary tract infection


Gene Reports ◽  
2021 ◽  
Vol 23 ◽  
pp. 101035
Author(s):  
Ali Akbar Shabani ◽  
Sima Ravaei ◽  
Hussein Anani ◽  
Reza Nasr ◽  
Mohammad Hussein Maroufi ◽  
...  

2003 ◽  
Vol 71 (6) ◽  
pp. 3088-3096 ◽  
Author(s):  
Peter Redford ◽  
Paula L. Roesch ◽  
Rodney A. Welch

ABSTRACT Extraintestinal Escherichia coli strains cause meningitis, sepsis, urinary tract infection, and other infections outside the bowel. We examined here extraintestinal E. coli strain CFT073 by differential fluorescence induction. Pools of CFT073 clones carrying a CFT073 genomic fragment library in a promoterless gfp vector were inoculated intraperitoneally into mice; bacteria were recovered by lavage 6 h later and then subjected to fluorescence-activated cell sorting. Eleven promoters were found to be active in the mouse but not in Luria-Bertani (LB) broth culture. Three are linked to genes for enterobactin, aerobactin, and yersiniabactin. Three others are linked to the metabolic genes metA, gltB, and sucA, and another was linked to iha, a possible adhesin. Three lie before open reading frames of unknown function. One promoter is associated with degS, an inner membrane protease. Mutants of the in vivo-induced loci were tested in competition with the wild type in mouse peritonitis. Of the mutants tested, only CFT073 degS was found to be attenuated in peritoneal and in urinary tract infection, with virulence restored by complementation. CFT073 degS shows growth similar to that of the wild type at 37°C but is impaired at 43°C or in 3% ethanol LB broth at 37°C. Compared to the wild type, the mutant shows similar serum survival, motility, hemolysis, erythrocyte agglutination, and tolerance to oxidative stress. It also has the same lipopolysaccharide appearance on a silver-stained gel. The basis for the virulence attenuation is unclear, but because DegS is needed for σE activity, our findings implicate σE and its regulon in E. coli extraintestinal pathogenesis.


PEDIATRICS ◽  
1964 ◽  
Vol 33 (6) ◽  
pp. 865-871
Author(s):  
Avron Y. Sweet ◽  
Emanuel Wolinsky

In a premature infant nursery 3 infants died of E. coli infections and 5 babies contracted urinary tract infection due to E. coli within a 5-month period. The causative organisms were found to be resistant to tetracycline, streptomycin, chloramphenicol, and sulfisoxazole, and sensitive to kanamycin, polymyxin B, and nitrofurantoin. From throat and stool cultures of 25 infants in the nursery, E. coli with this susceptibility pattern were recovered from 13. Of these, 9 were serotype 04:H5 and one was group 04 but nonmotile. Isolates from the urine of 3 infants with urinary tract infection and meninges of one fatal case were serotype 04:H5. Stool cultures revealed group 04 E. coli only in infants with that organism in the throat. The findings indicate that an outbreak of infections due to E. coli 04:H5 had occurred as a consequence of widespread colonization with that organism in a premature infant nursery. The unusual incidence of urinary tract infection due to a specific E. coli serotype indicates that urosepsis in the newborn can occur in epidemic form.


2013 ◽  
Vol 57 (11) ◽  
pp. 5197-5201 ◽  
Author(s):  
Katherine R. Ball ◽  
Francesca Sampieri ◽  
Manuel Chirino ◽  
Don L. Hamilton ◽  
Robert I. R. Blyth ◽  
...  

ABSTRACTA mouse model of cystitis caused by uropathogenicEscherichia coliwas used to study the distribution of gallium in bladder tissue following oral administration of gallium maltolate during urinary tract infection. The median concentration of gallium in homogenized bladder tissue from infected mice was 1.93 μg/g after daily administration of gallium maltolate for 5 days. Synchrotron X-ray fluorescence imaging and X-ray absorption spectroscopy of bladder sections confirmed that gallium arrived at the transitional epithelium, a potential site of uropathogenicE. coliinfection. Gallium and iron were similarly but not identically distributed in the tissues, suggesting that at least some distribution mechanisms are not common between the two elements. The results of this study indicate that gallium maltolate may be a suitable candidate for further development as a novel antimicrobial therapy for urinary tract infections caused by uropathogenicE. coli.


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