A Comparative Study of the Triphenyltetrazolium Chloride (Uroscreen) Test and Conventional Methods for the Detection of Bacteriuria

Acute urinary tract infection may be preceded by and active pyelonephritis may be associated with asymptomatic bacteriuria. Treatment of asymptomatic bacteriuria may prevent or arrest active, chronic pyelonephritis and its sequelae. Consequently, there is a need for a reliable and simple screening procedure to detect asymptomatic bacteriuria in large segments of the population.The reliability and practicability of tests advocated for the detection of bacteriuria, including the new chemical triphenyltetrazolium chloride (T.T.C.) (Uroscreen) test, were evaluated. Reliability was assessed by correlating results of these tests with bacterial counts of tested urines. Significant bacteriuria is defined as the presence of 100,000 or more organisms per ml. of urine.The T.T.C. (Uroscreen) test was positive in 92.5% of cases of bacteriuria; there were 7.5% false-negative and 2.8% false-positive results. Bacteria on Gram-stained smear were found in 95.5% of the cases of bacteriuria and in 14.6% of those with non-infected urine; pyuria (more than three leukocytes per high-power field), in 60% of those with bacteriuria and in 15.9% of those with presumably non-infected urine. Bacteria were conspicuous in the urinary sediment in 91.1% of cases of bacteriuria and in 3.7% of presumably non-infected urines.The T.T.C. (Uroscreen) test fulfilled the criteria for a reliable and simple screening procedure. It should be used concomitantly with other screening tests when the urine is examined routinely.     

Evaluation of microscopic examination for bacteriuria.

In order to find an easily available, simple, reliable and inexpensive method for demonstrating significant bacteriuria in routine urine examination, microscopic observation and bacteriological cultures have been made in parallel on total of 206 urine samples. Microscopic examinations of centrifuged deposit for both pus cells and bacteria were found to be more satisfactory in urine specimens with significant bacteriuria than the examinations for either of these elements alone. The criteria of more than five pus cells per high power field and organisms visible in methylene blue stain had sensitivity of 79% and a false positive rate of 13%.     

Neonatal Bacteriuria: A Prospective Study in 1,460 Infants

In a prospective study of 1,460 virtually consecutive neonates 14 (1%) were found to have bacteriuria, established by bladder puncture. Eleven were boys and three girls. Five of the infants with bacteriuria had symptoms suggestive of infection and were treated with antibiotics. The remaining nine were asymptomatic and remained so during a period of observation—five cleared their urine of bacteria without treatment, and four received antibiotics. Only one infant with bacteriuria had bacteraemia. Six infants had more than 10 white blood cells per mm³ in the bladder puncture urine and three had proteinuria. The blood urea was slightly raised in only two of the infants. The intravenous pyelogram was normal in all infants with bacteriuria but eight had slight or moderate vesicoureteric reflux when examined by micturating cystourethrography.     

Study of Childhood Urinary Tract Infection in General Practice

A study of bacteriuria was conducted among 426 of the 436 children under the age of 13 in a general practice in north-west London. Three girls and one boy were found to have asymptomatic bacteriuria, and a further girl with bacteriuria presented with abdominal pain and fever. The calculated incidence of urinary tract infection was 1·4% per annum. Most of the childhood urinary infections in this practice occurred before the age of 5 years, and the incidence of significant bacteriuria in this age group was 4·9% per annum. Five other children (four girls and one boy) in the practice were known to have had proved urinary tract infection. Of the total of eight children known to have had significant bacteriuria and investigated radiologically, three girls and two boys had radiological abnormalities in the urinary tract.Pyuria and proteinuria did not prove to be useful in the prediction of asymptomatic bacteriuria. Urinary tract infection with renal tract abnormality was found in this practice to be at least five times as common as diabetes in childhood.     

Incidence and outcome of symptomatic urinary tract infection in children.

The incidence of symptomatic urinary tract infection in 2879 children aged under 15 years was studied over 18 months in a single general practice. Infection was diagnosed if bacterial counts in three consecutive samples exceeded 100,000/ml. The incidence of urinary tract infection was 1.7 per 1000 boys at risk per year and 3.1 per 1000 girls. These values are lower than those of comparable studies, possibly because of the stricter diagnostic criterion used in the study. Urinary pus cell counts were also carried out and sometimes found to be misleading. Of the 14 children found to have an infection, three had a radiological abnormality. Five of the children had a recurrence of infection within the first two years, and one an asymptomatic bacteriuria seven years after diagnosis. Only six out of 34 children presenting with dysuria had infected urine, and an association was discovered between abacterial dysuria (or the urethral syndrome) in the remainder and a concurrent upper respiratory tract infection. All children should undergo radiological investigation after their first confirmed infection. Diagnosis and management could be improved by providing all general practitioners with a semiquantitative method of urine culture such as the dip slide.     

Adaptation of immunoenzymatic method for detection of antibody coated bacteria in urine

The modified immunoenzymatic method for detection of antibody coated bacteria (IP ACB) was compared with immunofluorescence technique (IF ACB) in the diagnosis of urinary tract infection. For the study 100 patients were employed with significant and insignificant bacteriuria. It was found that 81% of the results obtained by IP ACB and IF ACB were identical, however the immunoenzymatic method was more sensitive than immunofluorescence. Moreover, the IP ACB technique is simpler, less time consuming and may be performed by using the ordinary optic microscope.     

Trial evaluation of familial asymptomatic bacteriuria]

Between September 1, 1987 and December 1, 1988, 57 cases of asymptomatic bacteriuria were distinguished in the group of 2,898 schoolgirls, aged between 15 and 20 years (mean 16.7 years). For screening purposes Griess'test modified by Sleigh was used. Bacteriologic tests have shown, that 12 out 57 mothers (21.05%) and 5 out of 39 sisters (12.82%) suffered from asymptomatic bacteriuria, while this pathology was diagnosed in 61 (2.10%) out of all control population, i.e. 66 girls. Incidence of the asymptomatic bacteriuria was 4.55% in the 66 mothers. The differences in the incidence of the infection between the whole population and mothers of the control group, and between sisters and mothers of girls with asymptomatic bacteriuria were statistically significant (p < .001 and p < .01, respectively). History of the urinary tract infections was significantly more frequent in girls with asymptomatic bacteriuria, their mothers and sisters than in the control group. Type of bacterial strains isolated from the urine, similar sanitary conditions in these families as well as the lack of ultrasonographically and renographically diagnosed urinary pathologies in 11 out of 13 girls whose mothers and sisters suffered from asymptomatic bacteriuria suggest that familial asymptomatic bacteriuria may have a genetic background.     

Six year follow up of infants with bacteriuria on screening.

OBJECTIVE--To determine the value of screening for bacteriuria in infants with special emphasis on the natural course of untreated asymptomatic bacteriuria, renal growth, and renal damage. DESIGN--Prospective six year follow up of infants with bacteriuria on screening in an unselected infant population. SETTING--Paediatric outpatient clinic. PATIENTS--50 Infants (14 girls, 36 boys) with bacteriuria on screening verified by suprapubic aspiration from an unselected population of 3581 infants in a defined area of Gothenburg. INTERVENTIONS--Children with asymptomatic bacteriuria and normal findings on initial urography were untreated, although other infections were treated. MAIN OUTCOME MEASURES--Culture of urine and determination of C reactive protein concentration every six weeks for the first six months after diagnosis, every three months from six months to two years, and every six months between two and three years; thereafter yearly urine culture. Evaluation of renal concentrating capacity with a desmopressin test; radiological examination, including first and follow up urography and micturition cystourethrography without antibiotic cover; and measurement of renal parenchymal thickness and renal surface area. RESULTS--Of the original 50 infants, 37 (12 girls, 25 boys) were followed up for at least six years. Two infants developed pyelonephritis within two weeks after bacteriuria was diagnosed; the others remained free of symptoms. 45 Infants were untreated; the bacteriuria cleared spontaneously in 36 and in response to antibiotics given for infections in the respiratory tract in eight. Recurrences of bacteriuria were observed in 10 of the 50 children, of whom one had pyelonephritis. No child had more than one recurrence. At follow up urography in 36 of the 50 children (9 girls, 27 boys) after a median of 32 months no child had developed renal damage. First samples tested for renal concentrating capacity showed significantly higher values than those from a reference population (mean SD score 0.50, 95% confidence interval 0.21 to 0.79; p less than 0.001), but the last samples showed no significant difference (mean SD score 0.08, -0.24 to 0.40; p greater than 0.05). CONCLUSIONS--Mass screening for bacteriuria in infancy results primarily in detection of innocent bacteriuric episodes and is not recommended.     

Simple Disposable Method for Quantitative Cultures of Urine

A disposable kit was tested as a means of detecting significant bacteriuria by quantitative culture of urine. The total error in 3,563 specimens tested by five investigators was less than 1%. The method was very effective in differentiating significant bacteriuria, i.e., more than 100,000 bacteria per ml of urine from uninfected urine. In specimens from patients with urinary tract abnormalities who had mixed bacterial flora, the absolute numbers obtained with the dip-inoculum method had a 10% variation when compared to results obtained by calibrated loop or dilution pour plate methods. Therefore, the main utility of the kit is for screening and following patients after therapy. A significant delay in time between inoculation of the medium in the kit with the freshly voided urine and incubation of the kit to promote growth did not affect the reliability of the kit as a method of doing quantitative urine cultures to detect bacteriuria.     

Bacterial profile and drug susceptibility pattern of urinary tract infection in pregnant women at University of Gondar Teaching Hospital, Northwest Ethiopia

ABSTRACT: BACKGROUND: Urinary tract infection (UTI) is a common health problem among pregnant women. Proper investigation and prompt treatment are needed to prevent serious life threatening condition and morbidity due to urinary tract infection that can occur in pregnant women. Recent report in Addis Ababa, Ethiopia indicated the prevalence of UTI in pregnant women was 11.6 % and Gram negative bacteria was the predominant isolates and showed multi drug resistance. This study aimed to assess bacterial profile that causes urinary tract infection and their antimicrobial susceptibility pattern among pregnant women visiting antenatal clinic at University of Gondar Teaching Hospital, Northwest Ethiopia. METHODS: A cross-sectional study was conducted at University of Gondar Teaching Hospital from March 22 to April 30, 2011. Mid stream urine samples were collected and inoculated into Cystine Lactose Electrolyte Deficient medium (CLED). Colony counts yielding bacterial growth of 105/ml of urine or more of pure isolates were regarded as significant bacteriuria for infection. Colony from CLED was sub cultured onto MacConkey agar and blood agar plates. Identification was done using cultural characteristics and a series of biochemical tests. A standard method of agar disc diffusion susceptibility testing method was used to determine susceptibility patterns of the isolates. RESULTS: The overall prevalence of UTI in pregnant women was 10.4 %. The predominant bacterial pathogens were Escherichia coli 47.5 % followed by coagulase-negative staphylococci 22.5 %, Staphylococcus aureus 10 %, and Klebsiella pneumoniae 10 %. Gram negative isolates were resulted low susceptibility to co-trimoxazole (51.9 %) and tetracycline (40.7 %) whereas Gram positive showed susceptibility to ceftriaxon (84.6 %) and amoxicillin-clavulanic acid (92.3 %). Multiple drug resistance (resistance to two or more drugs) was observed in 95 % of the isolates. CONCLUSION: Significant bacteriuria was observed in asymptomatic pregnant women. Periodic studies are recommended to check the outcome of asymptomatic bacteriuria and also monitor any changes in the susceptibility patterns of urinary tract pathogens in pregnant women.     

13C-urea breath test to diagnose Helicobacter pylori infection in children aged up to 6 years

BACKGROUND: 13C-urea breath test (13C-UBT) is an accurate noninvasive tool for diagnosis of Helicobacter pylori infection. It is considered the best method for epidemiological studies, but there are few studies to evaluate the 13C-UBT in infants and toddlers. AIM: To evaluate the 13C-UBT performed with infrared spectroscopy in children aged up to 6 years. PATIENTS: Sixty-eight patients (6 months. to 5 years 11 months.) were evaluated prospectively and consecutively. METHODS: Helicobacter pylori infection was detected by positive culture, or rapid urease test and histological examination, both positive. 13C-UBT was performed with 50 mg of 13C-urea diluted in 100 ml of commercial orange juice. Two expired air samples were collected: before and 30 minutes after tracer ingestion. Cutoff of delta over baseline (DOB) was 4.0 per thousand and urea hydrolysis rate 10 microg/minute. RESULTS: Fifteen of 68 (22.1%) patients were H. pylori infected. Sensitivity was 93.3% (95% CI; 86.8%-99.7%) and specificity was 96.2% (95% CI; 93.6%-98.8%), and these values were equal for DOB and urea hydrolysis rate. Negative DOB values in noninfected patients ranged from -1.5 per thousand to 2.6 per thousand and positive DOB values ranged from 10.8 per thousand to 105.5 per thousand. There was no relationship between DOB values and age. Conclusion. 13C-UBT performed with infrared spectroscopy proved to be a reliable and accurate noninvasive diagnostic tool for H. pylori infection detection in children aged up to 6 years. Results far from cutoff value can clearly distinguish positive from negative 13C-UBT results in children up to 6 years old.     

Is screening for bacteriuria in pregnancy worth while?

A total of 4470 pregnant women were screened for bacteriuria by the dipslide method and significant growth found in 226 (5.1%). In 198 cases the urine was re-examined, in 119 by using suprapubic aspiration or catheterisation (62 (52%) samples contained bacteria) and in 79 by using midstream urine samples (26 (33%) samples contained greater than 10(8) colony forming units/1), showing the maximum prevalence of confirmed bacteriuria to be 2.6%. Overt urinary tract infection developed later in four of 80 patients with proved bacteriuria who had been given antibiotics, in one of eight untreated patients with bacteriuria, in one of 110 patients with unconfirmed bacteriuria, and in one of 226 non-bacteriuric controls. A history of urinary tract infection was given by 18% of controls and 42% of women with confirmed bacteriuria. Screening for bacteriuria and treatment with antibiotics to prevent later overt infection is expensive. Whether it is worth while and cost effective depends largely on the prevalence of bacteriuria in the local population and the proportion who develop overt infection. The screening and treatment programme reported here appeared to prevent only six cases of overt infection.     

Genetic determinants of Escherichia coli pathogenicity isolated from urine and feces of children with different clinical variants of urinary system infection

In the process of examination of 156 children of different age groups 176 E. coli cultures were isolated; of these, 98 cultures were isolated from acute cystitis and pyelonephritis patients, 28--from urine in cases of aysmptomatic bacteriuria, 30--from feces in cases of asymtomatc bacteriuria and intestinal dysbacteriosis, while 20 cultures--from feces of healthy children. In these bacteria the presence of genes associated with pathogenicity islets (PI) hlyA, hlyB, cnf-1, papC, sfaG and gene irp-2 (iron-regulated protein) was established with PCR. The detection rate of PI determinants in uropathogenic E. coli (UPEC) was shown to depend on the variants of the clinical manifestation of urinary tract infection. The total detection rate of PI gene fragments in UPEC cultures of different origin was indicative of their definitely less frequent occurrence in asymptomatic bacteriuria, observed simultaneously with intestinal dysbacteriosis, in comparison with acute urological infection. Practically the same detection rate of PI determinants in E. coli, isolated in asymptomatic bacteriuria in children, reflected high probability of genetic exchange in the above-mentioned fragments and made it possible to presume the existence of DNA sites, characteristic mainly of pathogenic clones. The established heterogeneity of the detection rate of PI determinants in E. coli clinical isolates requires further study.     

Evaluation of reagent strips in detecting asymptomatic bacteriuria in early pregnancy: prospective case series.

OBJECTIVE: To evaluate the performance of reagent test strips in screening pregnant women for asymptomatic bacteriuria at their first visit to an antenatal clinic. DESIGN: Prospective case series. SETTING: Antenatal clinic of a large inner city maternity hospital. SUBJECTS: All women attending for their first antenatal clinic. Patients taking antibiotics for any reason and those with urinary tract symptoms were excluded. INTERVENTION: A midstream urine specimen was divided; half was sent for microscopy and formal bacteriological culture and the other half was tested with a commercial reagent strip test for the presence of blood, protein, nitrite, and leucocyte esterase. MAIN OUTCOME MEASURES: Sensitivity, specificity, and positive and negative predictive values of the reagent strips in diagnosing asymptomatic bacteriuria (defined as 10(5) colony forming units/ml urine). RESULTS: Sensitivity was low, with a maximum of 33% when all four tests were used in combination. Specificity was high, with typical values of 99% or more. Positive predictive value reached a maximum of 69% and negative predictive value was typically 95% or more. CONCLUSION: Urine reagent strips are not sufficiently sensitive to be of use in the screening for asymptomatic bacteriuria and therefore many patients would be missed. In view of the potentially serious sequelae of this condition in pregnant women we recommend that formal bacteriological investigation remain the investigation of choice in this group of patients.     

Rapid screening for bacteriuria with a laser nephelometer

Detection of significant bacteriuria with a laser nephelometer was evaluated in this study and compared with the results obtained by the quantitative loop method. We screened 1002 urine specimens and 220 (21.95%) were found to be positive at greater than or equal to 10(5) colony-forming units (CFU)/mL of urine by the standard method. Of the 220 positive specimens, 210 (95.4%) were detected in 6 h or less and 177 (80.4%) were detected within 3 h. The false-positive rate was 2.3% at 3 h and 19.7% at 6 h. These findings suggest that a 6-h procedure is necessary to detect 95% or more of significant bacteriuria. Laser nephelometer is versatile and can be used for rapid screening of bacteriuria.     

Detection of Antibody-Coated Bacteria in Expectorated Sputum for Diagnosis of Lower Respiratory Infections

We evaluated antibody-coated bacteria (ACB) in expectorated sputum to discriminate contaminating or colonizing organisms from true pathogens. We examined 60 expectorated sputum samples from 51 patients with lower respiratory infections (chronic obstructive pulmonary disease 25, pneumonia 20, purulent tracheobronchitis 6). All samples were examined with quantitative culture and immunofluorescent demonstration of ACB. From the results of quantitative culture, we divided specimens into pathogen-isolated and pathogen-free samples. Among pathogen-isolated samples, in which we isolated accepted pathogenic organisms at ≥ 10⁷ colony-forming units per ml, 16 of 23 samples were ACB-positive (69.5%). In contrast, among pathogen-free samples, in which we isolated accepted pathogens at < 10⁷ colony forming units per ml or only upper respiratory flora, only 3 of 37 samples were ACB-positive (8.1%). The ACB-positive rate was significantly higher in pathogen-isolated than in pathogen-free samples (P < 0.001). Consequently, detecting ACB in expectorated sputum shows good potential as another criterion for distinguishing contaminating or colonizing organisms from true pathogens.     

Impact of a novel species of Nosema on the southwestern corn borer (Lepidoptera: Crambidae)

A study was undertaken to elucidate the impact of an undescribed Nosema sp. on the southwestern corn borer (SWCB; Diatraea grandiosella Dyar). The Nosema sp. (isolate 506) included in the study was isolated from an overwintering SWCB larva in Mississippi. It was highly infectious per os, with a median infective dose of 2.0 x 10(3) spores per larva. Even at the highest dosage tested (10(7) spores per larva), minimal mortality (< or = 3%) was observed in infected larvae, pupae, and adults reared in the laboratory on an artificial diet. However, infected pupae (0- and 7-d-old) were smaller, and the time to adult eclosion from pupation was slightly increased. Furthermore, the number of eggs produced by infected SWCB female moths substantially decreased (32%), and this effect was most pronounced on day 2, when the greatest number of eggs were oviposited by infected and noninfected moths. For eggs produced by infected females mated with infected males, hatch was slightly decreased by 16 and 15% for eggs laid on days 2 and 3, respectively. In addition, egg hatch was reduced in eggs oviposited by noninfected females mated with infected males on day 3. A low prevalence of infection (< 6%) was observed in the F1 generation originating from infected females mating with noninfected males, from noninfected females mating with infected males, and from infected females mating with infected males. Nosema 506 spores were observed in the proximity of reproductive tissues of infected female and male moths. Spores also were detected on the chorion surface and within eggs laid by infected females. Furthermore, 1-11% of larvae hatching from surface-sterilized eggs were infected by Nosema 506 indicating a transovarial mechanism of transmission.     

Untreated asymptomatic bacteriuria in girls: II--Effect of phenoxymethylpenicillin and erythromycin given for intercurrent infections.

OBJECTIVE--To investigate the effects of phenoxymethylpenicillin and erythromycin on urinary isolates from patients with untreated asymptomatic bacteriuria. DESIGN--Retrospective study of subgroup of patients from cohort followed up till the end of 1986. SETTING--Outpatient clinic for children with urinary tract infections. PATIENTS--51 Girls aged under 15 with untreated asymptomatic bacteriuria. INTERVENTIONS--Before 1982 intercurrent infections (mostly tonsillitis or otitis) were treated with phenoxymethylpenicillin; after 1982 erythromycin treatment was preferred. END POINTS--Change of bacterial strain in urinary tract and symptomatic recurrences of disease. MEASUREMENTS AND MAIN RESULTS--Bacteria identified by serotype and electrophoretic type and compared before and after antibiotic treatment. Bacteriuria eradicated and replaced by new strains in most patients treated with phenoxymethylpenicillin, leading to symptomatic recurrences in about 15%. Conversely, patients given erythromycin rarely showed change in bacteriuria and none suffered symptomatic recurrence. CONCLUSIONS--In girls with untreated asymptomatic bacteriuria the use of phenoxymethylpenicillin for intercurrent infections may lead to a change of urinary bacteria and leave them at substantial risk of acute pyelonephritis. With erythromycin this risk is small (2/20 courses in this series).     

Untreated asymptomatic bacteriuria in girls: I--Stability of urinary isolates.

OBJECTIVE--To assess the frequency of spontaneous changes of bacterial strains in patients with untreated asymptomatic bacteriuria. DESIGN--Retrospective analysis of samples from all patients with renal scarring and random sample of patients with normal kidneys. SETTING--Outpatient clinic for children with urinary tract infections. PATIENTS--54 Girls aged 3.3-15.5 years with untreated asymptomatic bacteriuria caused by Escherichia coli. INTERVENTION--None. END POINT--Change in bacterial strain. MEASUREMENTS AND RESULTS--Serotyping and electrophoretic analysis of sequential bacterial isolates, representing 151 patient years of untreated asymptomatic bacteriuria. A total of 24 changes of strain were identified. Eleven were related to medical interference such as treatment of other infections with antibiotics. CONCLUSIONS--Spontaneous changes of strain were uncommon, one change in 11.6 patient years, and thus are not a characteristic feature of the course of asymptomatic bacteriuria.     

Virulence of Escherichia coli in acute pyelonephritis,acute cystitis and asymptomatic bacteriuria

E. coli strains were isolated from urine specimens of hospitalised patients with acute pyelonephritis, acute cystitis or asymptomatic bacteriuria (ABU), and tested for virulence in an experimental mouse model. Of 12 pyelonephritisstrains 11 were shown to be virulent and 1 avirulent; of 12 cystitis-strains 4 were virulent and 8 avirulent; of 12 ABU-strains 5 were virulent and 7 avirulent. It is concluded that, while no difference in virulence was found between cystitis-and ABU-strains, pyelonephritis-strains were more often virulent than cystitis-and ABU-strains.No associations could be shown between virulence of the isolated strains and the presence of antibody-coated bacteria in the urine. Common urinary O types were not more often virulent than other O types. No relationship was seen between virulence and the presence of K antigen or the presence of particular K types.