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Low-count organisms concealed by dominant uropathogenic organisms in urine of patients with asymptomatic bacteriuria.

We compared organisms isolated by selective culture media with those isolated by routine bacteriological methods using the same urine specimens. Urine was obtained by suprapubic bladder aspiration from 84 patients suspected of having asymptomatic bacteriuria, including 31 catheterized patients. The selective media included mannitol salt agar, Enterococcus faecalis agar, nalidixic acid cetrimide agar, and desoxycholate hydrogen sulfide lactose agar with polymyxin B or cefsulodin. Colony counts on the selective media approximated to those on heart infusion agar, except for desoxycholate hydrogen sulfide agar with polymyxin B, where the number of organisms was underestimated. In 24 patients the selective media detected a total of 32 strains not reported by the hospital laboratory. The unreported strains included coagulase-negative staphylococci (41%), other cocci (38%), enterobacteriaceae (9%) and glucose-nonfermenting gram-negative bacilli (9%) and fungi (3%), with the number of organisms being mostly < 10(5)/ml. The majority of the unreported strains coexisted with dominant gram-negative bacilli or enterococci. These results indicate that low-count organisms coexisting with dominant organisms tend to be missed by the routine bacteriological methods.

Adolescent↗

The microbial flora of the vagina and its relationship to bacteriuria in diabetic and non-diabetic women.

The vaginal and urinary flora have been studied in 57 diabetic and 33 non-diabetic patients. In both populations, no correlation was found between urogenital symptoms and colonisation of the vagina. There was no greater prevalence of potentially pathogenic organisms in diabetic patients, even when control was poor. However, although not statistically significant, group B streptococci were isolated more frequently from poorly controlled diabetics. Bacteriuria was not significantly more common in diabetics. Gram-negative bacilli were found in the vagina significantly more often in bacteriuric patients; the prevalence of such organisms being four-fold greater after the menopause.

Adult↗

Diabetes mellitus in itself is not a risk factor for antibiotic resistance in Escherichia coli isolated from patients with bacteriuria.

AIM: To investigate whether diabetes mellitus is a risk factor for resistance in Escherichia coli isolated from patients with bacteriuria. METHODS: Data were obtained from a multicentre study. A clean-voided midstream urine culture was collected from 636 women with diabetes, who were between 18 and 75-years-old, attended an out-patient department and had no symptoms of a urinary tract infection. The resistance of E. coli was determined for different antimicrobials. The results were compared with resistance data from routine isolates of E. coli, obtained from women in the same age category, time period and location. RESULTS: A total of 135 E. coli were isolated from women with diabetes mellitus (mean age 57 +/- 14 years) and compared with 5907 routine isolates of E. coli obtained from female patients visiting an out-patient department (mean age 52 +/- 17 years). The resistance rates of E. coli isolated from diabetic patients and the routine isolates of E. coli to trimethoprim-sulfamethoxazole were 19% and 23%, respectively, to amoxicillin 16% and 32%, to nitrofurantoin 1% and 3%, to ciprofloxacin 0% and 4%, to ofloxacin 0% and 5%, and to norfloxacin 1% and 4%. CONCLUSION: The resistance of uropathogenic E. coli in non-hospitalized women with diabetes mellitus is not higher than that seen in routine isolates of E. coli. This suggests that diabetes in itself is not a risk factor for resistance.

Adolescent↗

Prevalence and risk factors for asymptomatic bacteriuria in women with Type 2 diabetes mellitus.

AIMS: Asymptomatic bacteriuria (ASB) has been considered as a complication in diabetic women. The reported data on the prevalence and various risk factors for ASB appear to be conflicting. Consequently, we investigated the prevalence and major risk factors of ASB in women with Type 2 diabetes mellitus. METHODS: A total of 411 non-pregnant women (aged 59.6 +/- 10.8 years) with Type 2 diabetes, and 160 women without diabetes (aged 53.3 +/- 15.1 years) assigned as controls, attending an outpatient endocrine clinic in a university-affiliated teaching hospital, were included. All participating women were interviewed and screened for the presence of ASB. In all participants, fasting blood glucose, HbA(1c) and renal function were measured. Complications of diabetes were also assessed. RESULTS: Of the 411 diabetic women, 25 (6.1%) had ASB, compared with four of 160 (2.5%) in control women (P = 0.07). Independent risk factors for the presence of ASB were albuminuria > 150 mg/24 h [odds ratio (OR) 4.96 (95% CI 1.64-15.0, P = 0.005)] and serum creatinine [OR 3.5 (95% CI 1.4-8.8, P = 0.008)]. No significant association was evident with age, BMI, duration of disease, glycaemic control assessed by HbA(1c) or chronic complications of diabetes, namely macrovascular disease, neuropathy and retinopathy. CONCLUSIONS: Women with Type 2 diabetes are not at higher risk of developing ASB than non-diabetic women. Independent and significant risk factors for ASB are macroalbuminuria and serum creatinine. The low prevalence of ASB found in this study may be as a result of the ethnic origin of these women and the circumcised state of their partners.

Adolescent↗

The Griess test: an inexpensive screening test for asymptomatic bacteriuria in pregnancy.

This study was designed to compare urine microscopy and the Griess test as screening tests for asymptomatic bacteriuria (AB) in pregnancy, using urine culture as a gold standard. Urine microscopy, Griess test and urine culture were carried out on urine samples from 483 antenatal women. AB was detected by urine culture in 30 (6.8%) of the patients. The Griess test and urine microscopy were found to have a sensitivity of 63.3% and 60%, specificity of 99.5% and 83.6%; and positive predictive value of 90.5% and 21.1% respectively. Most of the infections missed by the Griess test were either <100,000 colony count on culture or caused by gram positive cocci. The cost of the Griess test was only one tenth of that of urine microscopy. The Griess test is a simple and inexpensive test that when compared to urine microscopy has equal sensitivity, and better specificity and positive predictive value for the detection of AB in pregnancy than urine microscopy.

Bacteriological Techniques↗

Asymptomatic bacteriuria Escherichia coli strains: adhesins, growth and competition.

Urinary tract infections (UTIs) affect millions of people each year. Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU) in humans. Persons affected by ABU may carry a particular E. coli strain for extended periods of time without any symptoms. In contrast to uropathogenic E. coli (UPEC) that cause symptomatic UTI, very little is known about the mechanisms by which these strains colonize the urinary tract. Here, we have investigated the growth characteristics in human urine as well as adhesin repertoire of nine ABU strains; the ability of ABU strains to compete against the UPEC strain CFT073 was also studied. The different ABU strains displayed a wide variety of the measured characteristics. Half of the ABU strains displayed functional type 1 fimbriae while only one expressed functional P fimbriae. A good correlation between the growth rate of a particular strain and the survival of the strain in competition against CFT073 was observed. Our results support the notion that for strains with reduced capacity to express fimbriae, the ability to grow fast in human urine becomes crucial for colonization of the urinary tract.

Adhesins, Escherichia coli↗

Asymptomatic bacteriuria in school girls. IV Difficulties of level diagnosis and the possible relation to the character of infecting bacteria.

As no method for localization of urinary tract infection has been shown to be absolutely reliable, six methods have been run parallelly in a study of 60 school girls with asymptomatic bacteriuria caused by E. coli. A poor correlation was obtained between "high infection" indicated by bladder washout test and abnormal findings of C-reactive protein, sedimentation rate, antibody titres or renal concentrating capacity, while findings of parenchymal reduction on the pyelogram and reflux on the urethrocystogram were often found within this group. The low frequency of abnormal findings of C-reactive protein, sedimentation rate and renal concentrating capacity in girls with a "high infection" may be explained by the finding that the ABU patients were infected with changed E. coli strains probably not able to cause the usual inflammatory reactions. The patients' bacteria were found to be less efficient in providing antigen for antibody determinations than were standard strains of same O group, indicating a difference between strains from ABU patients and standard strains. From the battery of tests used it seemed that most of the girls had a bladder infection. All tests were normal in 48% of the patients while at least three of the methods were abnormal in 12%.

Adult↗

Evaluation of rapid methods for the detection of bacteriuria (screening) in primary health care.

The diagnostic performance of six methods for bacteriuria testing has been studied in 781 urine specimens obtained in primary health care, using conventional culture as reference method. The cut-off limits for classification of test results into positive and negative have been optimized with respect to diagnostic performance in primary health care. With optimized tests the following diagnostic efficiencies were obtained: Bacterial ATP, 0.94; Bacterial count in sediment, 0.93; Nitrite test, 0.92; Dipslide test, 0.92; White cell count in sediment; 0.87; Goffulocyte esterase test; 0.83. The diagnostic performance was also studied for all combinations of two tests. The highest diagnostic efficiency (0.96) was obtained by combining the ATP and dipslide tests. High diagnostic efficiencies can be obtained by a rapid primary test, using other tests for follow-up testing of specimens with intermediate or uninterpretable primary results. The most promising results were obtained by using ATP as the primary test, with follow-up testing of specimens with 3-25 nmol/l of ATP (12 per cent of the specimens). Follow-up testing by conventional culture resulted in overall diagnostic efficiency of 0.98. By performing the nitrite test on specimens with intermediary ATP-results, 81 per cent of the patients with UTI can be classified without culture. Only patients with intermediary ATP and negative nitrite results (10 per cent of the total number) will have to wait for final diagnosis based on conventional culture. Some alternative strategies to combine available methods are discussed in detail. Major advantages of the ATP test are that the test can be performed while the patients are waiting; it provides a numerical and objective result, and, in contrast to culture, it is not influenced by adhesion of bacteria to somatic cells.

Adenosine Triphosphate↗

Kinetics and dynamics of tobramycin action in patients with bacteriuria given single doses.

We studied the effect of a single intravenous dose of tobramycin on the rate of bacterial eradication from urine in 10 patients with bladder catheters. The catheter was replaced 4 to 6 h after the tobramycin dose. Pseudomonas aeruginosa was found in 7 of the 10 patients, while members of the family Enterobacteriaceae accounted for the remaining pathogens. The MIC for each bacterium was determined in both broth and urine. Tobramycin eradicated the bacteria from eight patients. Bacteriuria resolved in 21.8 +/- 18.0 h, and urine bactericidal activity persisted for 43.4 +/- 20.3 h after the dose of tobramycin. Most patients were recolonized by another bacterial species if use of Foley catheters was resumed on a continuous basis. Two patients required additional doses of tobramycin to eradicate the original pathogen. There were significant temporal relationships between the pharmacokinetics of tobramycin and the change in colony count of bacteria in urine.

Aged↗

Global gene expression profiling of asymptomatic bacteriuria Escherichia coli during biofilm growth in human urine.

Urinary tract infection (UTI) is an important health problem worldwide, with many millions of cases each year, and Escherichia coli is the most common organism causing UTI in humans. Also, E. coli is responsible for most infections in patients with chronic indwelling bladder catheter. The two asymptomatic bacteriuria (ABU) E. coli strains 83972 and VR50 are significantly better biofilm formers in their natural growth medium, human urine, than the two uropathogenic E. coli isolates CFT073 and 536. We used DNA microarrays to monitor the expression profile during biofilm growth in urine of the two ABU strains 83972 and VR50. Significant differences in expression levels were seen between the biofilm expression profiles of the two strains with the corresponding planktonic expression profiles in morpholinepropanesulfonic acid minimal laboratory medium and human urine; 417 and 355 genes were up- and down-regulated, respectively, during biofilm growth in urine of 83972 and VR50. Many genes involved in transcription and stress were up-regulated in biofilm-grown cells. The role in biofilm formation of four of the up-regulated genes, i.e., yceP, yqgA, ygiD, and aaeX, was investigated by creating single-knockout mutant versions of 83972 and VR50; all mutants showed reduced biofilm formation in urine by 18 to 43% compared with the wild type (P < 0.05). Also, the expression profile of strain 83972 in the human urinary tract partially overlaps with the biofilm expression profile.

Adaptation, Physiological↗

Global gene expression profiling of the asymptomatic bacteriuria Escherichia coli strain 83972 in the human urinary tract.

Urinary tract infections (UTIs) are an important health problem worldwide, with many million cases each year. Escherichia coli is the most common organism causing UTIs in humans. The asymptomatic bacteriuria E. coli strain 83972 is an excellent colonizer of the human urinary tract, where it causes long-term bladder colonization. The strain has been used for prophylactic purposes in patients prone to more severe and recurrent UTIs. For this study, we used DNA microarrays to monitor the expression profile of strain 83972 in the human urinary tract. Significant differences in expression levels were seen between the in vivo expression profiles of strain 83972 in three patients and the corresponding in vitro expression profiles in lab medium and human urine. The data revealed an in vivo lifestyle of microaerobic growth with respiration of nitrate coupled to degradation of sugar acids and amino acids, with no signs of attachment to host tissues. Interestingly, genes involved in NO protection and metabolism showed significant up-regulation in the patients. This is one of the first studies to address bacterial whole-genome expression in humans and the first study to investigate global gene expression of an E. coli strain in the human urinary tract.

Amino Acids↗

Interaction of human serum and neutrophils with Escherichia coli strains: differences between strains isolated from urine of patients with pyelonephritis or asymptomatic bacteriuria.

Strains of Escherichia coli isolated from the urine of children with pyelonephritis (n = 18) or asymptomatic bacteriuria (ABU) (n = 20) were serotyped and compared with respect to opsonic requirements, sensitivity to serum, amount of chemiluminescence produced by normal neutrophils during phagocytosis of the various strains, and amount of capsular antigen. Strains isolated from cases of ABU more often had the capacity to activate the alternative complement pathway than had strains causing pyelonephritis. The ABU strains were also better opsonized in fresh human serum and were more often serum sensitive than the pyelonephritic strains. There were individual variations in neutrophil chemiluminescence during phagocytosis of the various strains, but the two groups of strains did not differ significantly. The strains from pyelonephritis cases produced a greater amount of capsular antigens than the ABU strains. There was no correlation between presence of a particular O or K antigen and capacity to activate the alternative complement pathway. The observed differences between strains isolated from patients with ABU and those isolated from patients with pyelonephritis may be relevant for further studies on differences in bacterial virulence or host defense factors.

Antigens, Bacterial↗

Virulence properties of Escherichia coli 83972, a prototype strain associated with asymptomatic bacteriuria.

Little is known about bacteria associated with asymptomatic bacteriuria (ABU) with regard to urinary tract colonization mechanisms. In this study, virulence properties of Escherichia coli 83972, a strain that was isolated from a clinical ABU episode, were examined. The genetic potential for expression of P and type 1 pili was demonstrated, and DNA sequences related to type 1C and G (UCA) pilus genes were also detected. However, E. coli 83972 did not express D-mannose-resistant or D-mannose-sensitive hemagglutination after growth under standard conditions in vitro or upon isolation from the urine of colonized test subjects. Limited uroepithelial cell adherence was observed in vivo, and weak D-mannose-sensitive hemagglutination was detected after extended growth in urine in vitro.

Adhesins, Escherichia coli↗

The asymptomatic bacteriuria Escherichia coli strain 83972 outcompetes uropathogenic E. coli strains in human urine.

Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU). In contrast to uropathogenic E. coli (UPEC), which causes symptomatic urinary tract infections (UTI), very little is known about the mechanisms by which these strains colonize the human urinary tract. The prototype ABU E. coli strain 83972 was originally isolated from a girl who had carried it asymptomatically for 3 years. Deliberate colonization of UTI-susceptible individuals with E. coli 83972 has been used successfully as an alternative approach for the treatment of patients who are refractory to conventional therapy. Colonization with strain 83972 appears to prevent infection with UPEC strains in such patients despite the fact that this strain is unable to express the primary adhesins involved in UTI, viz. P and type 1 fimbriae. Here we investigated the growth characteristics of E. coli 83972 in human urine and show that it can outcompete a representative spectrum of UPEC strains for growth in urine. The unique ability of ABU E. coli 83972 to outcompete UPEC in urine was also demonstrated in a murine model of human UTI, confirming the selective advantage over UPEC in vivo. Comparison of global gene expression profiles of E. coli 83972 grown in lab medium and human urine revealed significant differences in expression levels in the two media; significant down-regulation of genes encoding virulence factors such as hemolysin, lipid A, and capsular polysaccharides was observed in cells grown in urine. Clearly, divergent abilities of ABU E. coli and UPEC to exploit human urine as a niche for persistence and survival suggest that these key differences may be exploited for preventative and/or therapeutic approaches.

Adhesins, Bacterial↗

Escherichia coli isolates associated with uncomplicated and complicated cystitis and asymptomatic bacteriuria possess similar phylogenies, virulence genes, and O-serogroup profiles.

The genetic and serological characteristics of Escherichia coli isolates from patients with uncomplicated cystitis (UC), complicated cystitis (CC), and complicated asymptomatic bacteriuria (CASB) were determined. Phylogenetic group B2 was predominant in all categories. The prevalences of 14 out of 18 virulence factor genes were similar among the three categories, while pap, iha, ompT, and PAI were more frequently seen in isolates associated with UC than CC or CASB.

Bacteriuria↗

Detection of bacteriuria by luciferase assay of adenosine triphosphate.

A selective method for distinguishing bacterial and nonbacterial adenosine triphosphate (ATP) in clinical bacteriological specimens was studied. The method involved incubation of samples with the detergent Triton X-100 and the ATP-hydrolyzing enzyme apyrase. The incubation selectively destroyed ATP in suspensions of various human cells while not affecting the ATP content in microbial cells. ATP remaining in the sample after incubation was extracted in boiling buffer and assayed by the firefly luciferase assay. Application of the method to 469 clinical urine specimens showed that the ATP level after treatment with Triton/apyrase was correlated to bacterial counts and that the sensitivity of the assay was sufficient for the detection of 10(5) bacteria/ml. The ATP levels per bacterial cell remaining in the urine specimen after treatment with Triton/apyrase were close to values observed in laboratory-grown cultures. The specificity and sensitivity of the luciferase assay for the detection of urinary bacteria and its possible use as a bacteriuria screening method are discussed.

Adenosine Triphosphate↗

Rapid screening for bacteriuria by light scatter photometry (Autobac): a collaborative study.

A total of 2,720 urine specimens from three laboratories were evaluated by Autobac (Pfizer Diagnostics) and were compared with simultaneous colony counts for evidence of bacteriuria. Of 599 specimens with a colony count of greater than or equal to 10(5) colony-forming units per ml, 93.8% were detected within 6 h. This detection rate increased to 97% of 447 positive urine specimens when only specimens from patients not on antimicrobials were evaluated. The majority (77.9%) of positive specimens were detected as early as 3 h. Those specimens with greater than or equal to 10(5) colony-forming units per ml, which were negative by Autobac at 6 h, included organisms which are frequently considered to be contaminants (diphtheroids, lactobacilli, alpha and gamma streptococci, yeasts, and Staphylococcus epidermidis), or were from patients who were being treated with antimicrobial agents. Of 2,121 urine specimens with colony counts of less than 10(5), 98.1% were correctly determined to be negative by Autobac at 3 h. This percentage decreased to 86.0 at 6 h. The majority of these false-positive specimens were those with colony counts of 10(4) to 10(5) colony-forming units per ml. There appeared to be no major difference in results from the three laboratories or among the four lots of broth used in this study.

Anti-Bacterial Agents↗

Colorimetric method for rapid determination of bacteriuria.

An inexpensive, rapid, and simple colorimetric test for detection of bacteriuria is described. This test does not require bacterial growth and has the marked advantage of being able to quantify bacteria, even when the organisms are present in the urine of bacteriuric patients who are being treated with antibiotics. The test is carried out with 1 ml of urine, which is processed through a 10-mm-diameter filter than entraps the bacteria on its surface. Safranine dye is passed through the filter to stain the bacteria and the filter fibers. A decolorizer, which removes the dye from the filter fibers but not from the bacteria, is then passed through the filter. If there are greater than or equal to 10(5) colony-forming units of bacteria per ml in the sample, the 10-mm filter disk manifests a pink to red color. If there are less than 10(5)colony-forming units of bacteria per ml, the filter disk remains white or becomes slightly yellow. The entire procedure has been adapted to a semiautomated instrument and the time required per test is less than 1 min. The results obtained on the test card are a permanent record to be filled with the patient's chart. The bacteria can be quickly classified as gram positive or gram negative by selective staining of a second milliliter of urine on the filter. Of 441 urine specimens tested, 430 (98%) were correctly classified as containing more or less than 10(5) colony-forming units per ml. A total of 62 urine specimens were positive by bacterial plating (greater than or equal to 10(5) colony-forming units per ml), and 59 were positive by the colorimetric test. Eight false-positives (colorimetric test positive, plate counts less than 10(5)) were encountered in patients (bacteriuric) being prescribed antibiotics. Removal of the antibiotics from these urine specimens, with subsequent replating of the samples, indicated the presence of greater than or equal to 10(5) colony-forming units of bacteria per ml in three representative cases tested, indicating that the results of the colorimetric tests were not false-positives but that the plate counts were low because of the inhibition of bacterial growth by the residual antibiotics present in the urine.

Bacteria↗