Pyelonephritis Isolates Research Articles

Comparative study of Escherichia coli virulence determinants in strains causing urinary tract bacteremia versus strains causing pyelonephritis and other sources of bacteremia

To assess the role of phylogenetic background and putative virulence factors (VFs) in Escherichia coli causing urinary bacteremia, 50 strains isolated from this condition were compared with 50 strains isolated from pyelonephritis and 50 from other sources of bacteremia. papA and papGII were significantly more prevalent in urinary bacteremia and pyelonephritis (78%, 66% and 70%, 58%) than in other-source bacteremia (48% and 24%), whereas sfa/focDE and cnf1 were more prevalent in urinary-source bacteremia (56% and 44%) than in pyelonephritis and other-source bacteremia (28%, 42% and 20%, 28%). Group B2 was the most frequent in all conditions (63% of isolates) and exhibited the greatest concentration of VFs. Urinary tract bacteremia, pyelonephritis, and other-source bacteremia isolates presented similar virulence scores (7.8, 7.0, and 6.6); however, there were striking differences among the phylogenetic groups (8.7 in group B2 versus 3.4 in group A; P < .001). Group A and B1 strains almost exclusively infected compromised hosts.

ESCHERICHIA COLI STRAINS CAUSING URINARY TRACT INFECTION IN UNCIRCUMCISED INFANTS RESEMBLE UROSEPSIS-LIKE ADULT STRAINS

Urinary tract infection (UTI) is the most frequent bacterial infection in infants younger than 90 days, and mainly affects uncircumcised males. In an attempt to unravel further the pathophysiology of UTI in this age group we used molecular methods to characterize Escherichia coli strains responsible for community acquired UTI in male and female infants and in adults. A total of 79 E. coli isolates from uncircumcised male and female infants with UTI and no urinary tract abnormalities and 41 E. coli pyelonephritis isolates from nonhost compromised adults with bacteremia were characterized in terms of phylogenic relatedness and virulence factors. Males were infected by strains with a genetic background and virulence factors different from those affecting females but similar to those of adult pyelonephritis strains. Our molecular approach indicates that uncircumcised male infants are at higher risk for infection with highly virulent uropathogenic E. coli strains than are females. Preputial colonization may have a key role in the selection of such strains.

Extended Virulence Genotypes and Phylogenetic Background ofEscherichia coliIsolates from Patients with Cystitis, Pyelonephritis, or Prostatitis

Molecular analysis of 63 Escherichia coli urine isolates showed that pyelonephritis (n=23) and prostatitis (n=17) isolates exhibited more virulence factors (VFs) among the 35 sought than did cystitis isolates (n=23). Several nontraditional VFs--including bmaE (M fimbriae), gafD (G fimbriae), fyuA (yersiniabactin receptor), ireA and iroN (novel siderophore receptors), cvaC (colicin [microcin] V), traT (serum-resistance associated), ibeA (invasion of brain endothelium), ompT (outer membrane protease T), and malX (pathogenicity island marker)--either differentiated significantly between syndromes (despite small numbers of isolates and possible multiple-comparison artifacts) or were broadly prevalent. Thus, interventions that target conserved uro-VFs may be possible, despite the likely existence of syndrome-specific pathogenetic mechanisms and/or host defense systems.

Osmoregulatory systems of Escherichia coli: identification of betaine-carnitine-choline transporter family member BetU and distributions of betU and trkG among pathogenic and nonpathogenic isolates.

Multiple transporters mediate osmoregulatory solute accumulation in Escherichia coli K-12. The larger genomes of naturally occurring strains such as pyelonephritis isolates CFT073 and HU734 may encode additional osmoregulatory systems. CFT073 is more osmotolerant than HU734 in the absence of organic osmoprotectants, yet both strains grew in high osmolality medium at low K(+) (micromolar concentrations) and retained locus trkH, which encodes an osmoregulatory K(+) transporter. Both lacked the trkH homologue trkG. Transporters ProP and ProU account for all glycine-betaine uptake activity in E. coli K-12 and CFT073, but not in HU734, yet elimination of ProP and ProU impairs the growth of HU734, but not CFT073, in high osmolality human urine. No known osmoprotectant stimulated the growth of CFT073 in high osmolality minimal medium, but putative transporters YhjE, YiaMNO, and YehWXYZ may mediate uptake of additional osmoprotectants. Gene betU was isolated from HU734 by functional complementation and shown to encode a betaine uptake system that belongs to the betaine-choline-carnitine transporter family. The incidence of trkG and betU within the ECOR collection, representatives of the E. coli pathotypes (PATH), and additional strains associated with urinary tract infection (UTI) were determined. Gene trkG was present in 66% of the ECOR collection but only in 16% of the PATH and UTI collections. Gene betU was more frequently detected in ECOR groups B2 and D (50% of isolates) than in groups A, B1, and E (20%), but it was similar in overall incidence in the ECOR collection and in the combined UTI and PATH collections (32 and 34%, respectively). Genes trkG and betU may have been acquired by lateral gene transfer, since trkG is part of the rac prophage and betU is flanked by putative insertion sequences. Thus, BetU and TrkG contribute, with other systems, to the osmoregulatory capacity of the species E. coli, but they are not characteristic of a particular phylogenetic group or pathotype.

A disseminated multidrug-resistant clonal group of uropathogenic Escherichia coli in pyelonephritis

In acute pyelonephritis, bacterial resistance to co-trimoxazole predicts treatment failure, but the clonal basis of such resistance is undefined. We did molecular and serological analyses of 170 Escherichia coli urine isolates obtained in 1994-96 from women with acute pyelonephritis. 12 (7%) of the pyelonephritis isolates were in clonal group A (CGA; responsible for 38-51% of co-trimoxazole resistance in acute cystitis), including ten (34%) of 29 isolates that were resistant to co-trimoxazole. CGA isolates were obtained from diverse locations across the USA and were related to the O15:K52:H1 clone of the 1986-87 outbreak in London, UK. Thus, CGA is broadly disseminated and contributes to co-trimoxazole resistance in pyelonephritis as well as in cystitis.

Identification of DNA sequences from a second pathogenicity island of uropathogenic Escherichia coli CFT073: probes specific for uropathogenic populations.

Uropathogenic Escherichia coli is the leading cause of urinary tract infection and hospital visits in North America. Cystitis and acute pyelonephritis, infection of the bladder and kidney, respectively, are the two most common syndromes encountered in patients with urinary tract infection. We sequenced and annotated 71,684 bases of a previously unidentified pathogenicity-associated island (PAI) from E. coli strain CFT073. This PAI contained 89 open-reading frames encoding a pap operon, iron-regulated genes, mobile genetic elements, and a large proportion of unknown or unidentified open-reading frames. Dot blot analysis with 11 DNA sequences from this PAI demonstrated that 7 sequences were more prevalent among uropathogens: 2 probes were more prevalent among cystitis and pyelonephritis isolates, 2 among pyelonephritis isolates only, and 3 among cystitis isolates only than among fecal isolates. These data suggest that groups of uropathogens have genetic differences that may be responsible for the different clinical outcomes.

Dr operon-associated invasiveness of Escherichia coli from pregnant patients with pyelonephritis.

We used a gentamicin protection assay to assess the ability of gestational pyelonephritis isolates of Escherichia coli to invade HeLa cells. The ability to enter HeLa cells was strongly associated with the presence of Dr operons coding for Dr adhesins. In contrast, the nonivasive isolates predominantly expressed papG, coding for P fimbriae.

The osmotic stress response and virulence in pyelonephritis isolates of Escherichia coli: contributions of RpoS, ProP, ProU and other systems.

Trehalose synthesis (RpoS-dependent) and betaine uptake mediated by transporters ProP and ProU contribute to the osmotolerance of Escherichia coli K-12. Pyelonephritis isolates CFT073 and HU734 were similar and diminished in osmotolerance, respectively, compared to E. coli K-12. The roles of RpoS, ProP and ProU in osmoregulation and urovirulence were assessed for these isolates. Strain HU734 expressed an RpoS variant which had low activity and a C-terminal extension. This bacterium accumulated very little trehalose and had poor stationary-phase thermotolerance. For E. coli CFT073, introduction of an rpoS deletion impaired trehalose accumulation, osmotolerance and stationary-phase thermotolerance. The rpoS defects accounted for the difference in osmotolerance between these strains in minimal medium of very high osmolality (1.4 mol kg(-1)) but not in medium of lower osmolality (0.4 mol kg(-1)). The slow growth of both pyelonephritis isolates in high-osmolality medium was stimulated by glycine betaine (GB) and deletion of proP and/or proU impaired GB uptake. An HU734 derivative lacking both proP and proU retained osmoprotective GB uptake activity that could be attributed to system BetU, which is not present in strain K-12 or CFT073. BetU transported GB (K(m), 22 microM) and proline betaine. High-osmolality human urine (0.92 mol kg(-1)) included membrane-permeant osmolyte urea (0.44 M) plus other constituents which contributed an osmolality of only approximately 0.4 mol kg(-1). Strains HU734 and CFT073 showed correspondingly low GB uptake activities after cultivation in this urine. Deletion of proP and proU slowed the growth of E. coli HU734 in this high-osmolality human urine (which contains betaines) but had little impact on its colonization of the murine urinary tract after transurethral inoculation. By contrast, deletion of rpoS, proP and proU had no effect on the very rapid growth of CFT073 in high-osmolality urine or on its experimental colonization of the murine urinary tract. RpoS-dependent gene expression is not essential for growth in human urine or colonization of the murine urinary tract. Additional osmoregulatory systems, some not present in E. coli K-12 (e.g. BetU), may facilitate growth of pyelonephritis isolates in human urine and colonization of mammalian urinary tracts. The contributions of systems ProP and ProU to urinary tract colonization cannot be definitively assessed until all such systems are identified.

In vivo dynamics of type 1 fimbria regulation in uropathogenic Escherichia coli during experimental urinary tract infection.

Escherichia coli is the primary cause of uncomplicated infections of the urinary tract including cystitis. More serious infections, characterized as acute pyelonephritis, can also develop. Type 1 fimbriae of E. coli contribute to virulence in the urinary tract; however, only recently has the expression of the type 1 fimbriae been investigated in vivo using molecular techniques. Transcription of type 1 fimbrial genes is controlled by a promoter that resides on a 314-bp invertible element capable of two orientations. One places the promoter in the ON orientation, allowing for transcription; the other places the promoter in the OFF orientation, preventing transcription. A PCR-based assay was developed to measure the orientation of the invertible element during an experimental urinary tract infection in mice. Using this assay, it was found that the percentage of the population ON in urine samples correlated with the respective CFU per gram of bladder (P = 0.0006) but not with CFU per gram of kidney (P > 0.069). Cystitis isolates present in the urine of mice during the course of infection had a higher percentage of their invertible elements in the ON orientation than did pyelonephritis isolates (85 and 34%, respectively, at 24 h; P < 0.0001). In general, cystitis isolates, unlike pyelonephritis isolates, were more likely to maintain their invertible elements in the ON orientation for the entire period of infection. E. coli cells expressing type 1 fimbriae, expelled in urine, were shown by scanning electron microscopy to be densely packed on the surface of uroepithelial cells. These results suggest that expression of type 1 fimbriae is more critical for cystitis strains than for pyelonephritis strains in the early stages of an infection during bladder colonization.

Ampicillin-resistant Escherichia coli in gestational pyelonephritis: increased occurrence and association with the colonization factor Dr adhesin.

The pattern of ampicillin resistance and possible association with virulence factors of 78 Escherichia coli isolates taken from 78 pregnant women with pyelonephritis were evaluated. The current incidence of ampicillin resistance among pyelonephritis isolates (46%) was significantly higher than that reported in 1985 (22%). Resistance was found more frequently during the first (60%) and third (53%) trimesters than during the second trimester (33%). Of all dra(+) E. coli isolates, 75% were ampicillin resistant, whereas dra(+) isolates of O75 serotype E. coli accounted for 87% of ampicillin-resistant strains. The significant increase of ampicillin resistance among gestational pyelonephritis E. coli and the association with the dra gene cluster encoding colonization and invasive capacity may warrant further study involving obstetric and neonate wards, with the latter being at the higher risk for potential problems.

Virulence characteristics of Escherichia coli in acute bacterial prostatitis.

To assess the urovirulence characteristics of Escherichia coli strains causing acute prostatitis, urinary isolates from men with acute prostatitis (n=107) and from women with acute uncomplicated pyelonephritis (n=76) were examined for the prevalence of sfa, foc, and 3 papG allele genotypes and phenotypes and for the production of alpha-hemolysin and cytotoxic necrotizing factor 1. The papG allele III and foc gene were found more frequently and the papG allele II less frequently among prostatitis than from pyelonephritis isolates. A higher proportion of hly+ cnf1+ genotype in prostatitis strains (64% vs. 36%) was particularly striking. Both prostatitis and pyelonephritis strains expressed virulence factors similarly except for a higher proportion of nonhemolytic prostatitis isolates. Although the pathogenetic mechanisms of urinary tract infections in men and women may differ, virulence factors such as adhesins and cytotoxins may have important roles in the pathogenesis of acute prostatitis.

Osmoregulatory transporter ProP influences colonization of the urinary tract by Escherichia coli.

Osmoregulatory transporters ProP and ProU mediate the use of betaines as osmoprotectants by Escherichia coli. Glycine betaine and proline betaine are present in mammalian urines. Betaine uptake may therefore facilitate the growth of E. coli in the urinary tract, an environment of fluctuating osmolality. ProP transporter activity was approximately threefold higher in a pyelonephritis isolate, E. coli HU734, than in E. coli K-12. The growth rate of E. coli HU734 in aerated minimal salts medium was reduced twofold by 0.2 M NaCl in the absence and by 0.55 M NaCl in the presence of glycine betaine. Maximal growth rate stimulation was achieved when glycine betaine was added at a concentration as low as 25 microM. Deletion of the proP locus impaired the growth rate of E. coli HU734 in human urine but not in minimal medium supplemented with NaCl (0.4 M), with or without glycine betaine (0.1 mM). The expression of pyelonephritis-associated (P) pili was reduced when E. coli HU734 was cultured in a rich culture medium (LB) of elevated salinity. The proP lesion had no influence on P pilus expression in vitro or on the recovery of bacteria from the kidneys of inoculated mice. However, it did reduce their recovery from the bladders of inoculated mice 100-fold. These data provide the first direct evidence that osmoprotective betaine accumulation and transporter ProP are pertinent to both growth in human urine and colonization of the murine urinary tract by uropathogenic E. coli.

Internalization of Escherichia coli by human renal epithelial cells is associated with tyrosine phosphorylation of specific host cell proteins

Human renal epithelial cells are capable of internalizing Escherichia coli regardless of whether the bacteria are isolated from individuals with pyelonephritis or from healthy volunteers. In this study, we investigated the role of host cell tyrosine kinase activity in internalization. We found that internalization of both fecal and pyelonephritis isolates is blocked by tyrosine kinase inhibitors. We found increased intensity of two tyrosine-phosphorylated proteins, with relative mobilities of approximately 123,000 and 110,000, in Western blots of extracts from human renal epithelial cells infected with E. coli. The increased intensity of these tyrosine-phosphorylated proteins was observed only in the Triton X-100-insoluble fraction, suggesting that these proteins could be associated with the cytoskeleton. Increased tyrosine phosphorylation of these proteins upon E. coli infection was observed in both transformed and primary human renal epithelial cells and in cells infected with several different strains of E. coli isolated from the feces of healthy individuals or from the blood or urine of patients with pyelonephritis. The increased tyrosine phosphorylation of these proteins required live bacteria and was blocked by tyrosine kinase inhibition but not by protein synthesis inhibitors or cytochalasin D. These experiments establish a strong link between E. coli internalization and host cell signaling through tyrosine kinases in human kidney cells and provide evidence that specific proteins are involved in these processes.

Escherichia coli virulence factors and serotypes in acute bacterial prostatitis.

Escherichia coli is the most frequent pathogen in both acute bacterial prostatitis and acute uncomplicated urinary infections. To assess the virulence profiles of E. coli in acute prostatitis, the serotypes and virulence factor (VF) genotypes were determined. We studied 107 E. coli isolates from cases of acute bacterial prostatitis, 76 isolates from acute pyelonephritis, 194 isolates from acute Cystitis and 80 fecal isolates from healthy people. All pyelonephritis and cystitis isolates were from women. Seven urovirulence determinants were analyzed by DNA colony hybridization, including the genes for type 1 fimbria (pil), P fimbria (pap), S fimbria (sfa), afimbrial adhesin AFA-I (afaI), alpha-hemolysin (hly), cytotoxic necrotizing factor 1 (cnf1) and aerobactin (aer). O:H:K serotypes were also determined. With the exception of pil and afaI, all VFs were significantly more often associated with prostatitis, pyelonephritis and cystitis isolates than with the fecal isolates. The prevalence of sfa, hly and cnf1 was higher in prostatitis isolates than in pyelonephritis and cystitis isolates, and the pap+sfa+hly+cnf1+ genotype was dominant among prostatitis isolates (48.8%). Nine O serotype (O1, O2, O4, O6, O16, O18, O22, O25 and O75) accounted for 79.4%, 73.7% and 78.4% of the prostatitis, pyelonephritis and cystitis strains, respectively. There was an apparent correlation between serotype and genotype in uropathogenic E. coli. The predominance of O serotypes in female urinary tract infections and a high percentage of multiple VFs among the prostatitis isolates suggested that VFs play important roles in the pathogenesis of acute bacterial prostatitis.

Dra-related X adhesins of gestational pyelonephritis-associated Escherichia coli recognize SCR-3 and SCR-4 domains of recombinant decay-accelerating factor

Bacterial adhesins are important virulence factors that allow colonization of the human urogenital tract by Escherichia coli. Adhesins of the Dr family have been found to be more frequently expressed in strains associated with symptomatic urinary tract infections. Because of the high frequency of symptomatic urinary tract infections during pregnancy, we screened E. coli isolates from 64 gestational pyelonephritis patients for the expression of Dr and X adhesins to address their potential virulence roles in this population. Using PCR and primers for the afaB gene, we detected dra-related operons in 17 isolates (27%). On the basis of the lack of hemagglutination of Dr(a-) erythrocytes containing a point mutation in the decay-accelerating factor (DAF) short consensus repeat-3 (SCR-3) domain, 12 of these strains were categorized as classical Dr adhesins. The hemagglutination of O erythrocytes by Dr+ strains was blocked or reduced by a monoclonal antibody to the DAF SCR-3 domain. The remaining five dra-positive strains agglutinated Dr(a-) erythrocytes. Monoclonal antibody to the DAF SCR-3 domain failed to block O-erythrocyte hemagglutination. Adhesins in these strains did not fulfill criteria for Dr hemagglutinins because of the undefined receptor specificities and were categorized as X. E. coli strains bearing dra-related X adhesins bound to DAF cDNA-transfected Chinese hamster ovary cells. Three of these dra-related X-adhesin-bearing E. coli strains failed to attach to the SCR-3 delta deletion transfectant, which suggested that binding sites were located in the SCR-3 domain but outside the region blocked by the monoclonal anti-SCR-3 immunoglobulin G. The binding sites of the remaining two dra-related X adhesin strains were localized to the SCR-4 domain, as the attachment was shown to be abolished on an SCR-4 delta mutant but unaffected by an SCR-3 delta deletion. The heterogeneity in the binding sites of E. coli DAF (Dr) family adhesins from gestational pyelonephritis isolates may reflect the ability of the adhesins to evolve to recognize alternate peptide epitopes for efficient colonization.

Internalization of Escherichia coli into human kidney epithelial cells: comparison of fecal and pyelonephritis-associated strains.

A gentamicin survival assay, using primary human renal epithelial cells and Escherichia coli strains isolated from the feces of asymptomatic individuals and from the urine or blood of patients with acute pyelonephritis, was used to investigate bacterial internalization as a model for renal parenchymal invasion in pyelonephritis. E. coli strains, regardless of their origin, efficiently entered into human renal epithelial cells, a process inhibited by cytochalasin D. While the percentage of survival of nonhemolytic pyelonephritis isolates did not differ from that of fecal isolates, survival of hemolytic pyelonephritis strains was lower than that of nonhemolytic strains, perhaps as a consequence of the greater cytotoxicity of hemolytic strains. There was no evidence of intracellular multiplication of E. coli. These results demonstrate that human renal epithelial cells are capable of efficient uptake of E. coli regardless of the source of the bacteria.

Serotypes, hemagglutinins, siderophore synthesis, and serum resistance of Klebsiella isolates causing human urinary tract infections.

A total of 146 Klebsiella isolates from human asymptomatic bacteriuria (n = 73), cystitis (n = 54), and acute pyelonephritis (n = 19) were examined for the presence of particular virulence factors. Capsular type K2 was the most common serotype observed (13%). This capsule type was prevalent in isolates from asymptomatic bacteriuria and cystitis but not from pyelonephritis. Type 1 fimbriae were found significantly more often in pyelonephritis isolates than among those from asymptomatic and symptomatic lower urinary tract infection (UTI; P < .05), while no marked differences were detected with respect to the distribution of type 3 fimbriae. Serum resistance was more frequent among isolates from symptomatic (26%) than from asymptomatic UTI (18%). Enterochelin was produced by all but 1 of the isolates as determined by a bioassay. In contrast, aerobactin synthesis was rare (3%), with isolates from pyelonephritis showing the highest frequency of aerobactin production (3/19).

Proteus mirabilis flagella and MR/P fimbriae: isolation, purification, N-terminal analysis, and serum antibody response following experimental urinary tract infection

Urinary tract infection with Proteus mirabilis may lead to serious complications, including cystitis, acute pyelonephritis, fever, bacteremia, and death. In addition to the production of hemolysin and the enzyme urease, fimbriae and flagellum-mediated motility have been postulated as virulence factors for this species. We purified mannose-resistant/proteuslike (MR/P) fimbriae and flagella from strains CFT322 and HU2450, respectively. Electron microscopy revealed highly concentrated preparations of fimbriae and flagella. Fimbrial and flagellar structural subunits were estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 18.5 and 41 kDa, respectively. N-terminal sequencing revealed that 10 of the first 20 amino acids of the major MR/P subunit matched the sequence of the P. mirabilis uroepithelial cell adhesin N terminus and 11 of 20 amino acids matched the predicted amino acid sequence of the Escherichia coli P fimbriae structural subunit, PapA. In addition, 90 and 80% homologies were found between the first 20 amino acids of P. mirabilis flagellin and those of Salmonella typhimurium phase-1 flagellin and the E. coli hag gene product, respectively. An enzyme-linked immunosorbent assay using purified antigens showed a strong reaction between the MR/P fimbriae or flagella and sera of CBA mice challenged transurethrally with P. mirabilis. A possible role for MR/P fimbriae in the pathogenesis of urinary tract infection is supported by (i) a strong immune response to the antigen in experimentally infected animals, (ii) amino acid sequence similarity to other enteric surface structure, and (iii) our previously reported observation that MR/P fimbriae are expressed preferentially as the sole fimbrial type in human pyelonephritis isolates.

Frequency and Organization of pap Homologous DNA in Relation to Clinical Origin of Uropathogenic Escherichia coli

The pap operon encodes the gal alpha 1-4gal beta specific adhesins of Escherichia coli. The presence and organization of pap homologous DNA was determined using two probes specific for pap in 217 uropathogenic E. coli samples by dot blot and Southern blot analysis. The frequency of pap homologous DNA was 76% in pyelonephritis, 69% in cystitis, and 52% in an asymptomatic bacteriuria group. Further, the gal alpha 1-4gal beta binding phenotype among the pap-positive strains was expressed more often in acute pyelonephritis (91%) than the cystitis (60%) or asymptomatic bacteriuria (52%) strains. This was explained in part by difference in organization of pap homologous DNA between the genotypically positive pyelonephritis and asymptomatic bacteriuria strains. The pyelonephritis isolates contained three copies of pap significantly more often than the asymptomatic bacteriuria strains, and the pyelonephritogenic O-antigen types had a general increase in pap copy number. The difference in expression of gal alpha 1-4gal beta adhesins between pyelonephritis and asymptomatic bacteriuria isolates was thus not only a function of the frequency of pap homologous DNA but also of phenotypic expression among genotypically pap-positive strains.

Hemagglutinin, Urease, and Hemolysin Production by Proteus mirabilis from Clinical Sources

Proteus mirabilis, a common cause of urinary tract infection, can lead to serious complications including pyelonephritis. Adherence factors, urease, and hemolysin may be virulence determinants. These factors were compared for bacteria cultured from 16 patients with acute pyelonephritis and 35 with catheter-associated bacteriuria and for 20 fecal isolates. Pyelonephritis isolates were more likely (P less than .05) to express the mannose-resistant/Proteus-like (MR/P) hemagglutinin in the absence of mannose-resistant/Klebsiella-like (MR/K) hemagglutinin than were catheter-associated or fecal isolates. Pyelonephritis isolates produced urease activity of 63 +/- 27 (mean +/- SD) mumol of NH3/min/mg of protein, not significantly different from catheter-associated or fecal isolates. Hybridization of Southern blots of P. mirabilis chromosomal DNA with two urease gene probes demonstrated that urease gene sequences were conserved in all isolates. Geometric mean of reciprocal hemolytic titers for pyelonephritis isolates was 27.9; for urinary catheter isolates, 18.0; and for fecal isolates, 55.7 (not significantly different, P greater than .1). Although in vivo expression of urease and hemolysin may not be reliable indexes of virulence, MR/P hemagglutination in the absence of MR/K hemagglutination may be necessary for development of pyelonephritis.