Gerald V. Ling

Evidence of Commonality between Canine and Human Extraintestinal Pathogenic Escherichia coli Strains That Express papG Allele III

ABSTRACT Although dogs have been proposed as carriers of extraintestinal pathogenic Escherichia coli (ExPEC) with infectious potential for humans, presumed host species-specific differences between canine and human ExPEC strains have cast doubt on this hypothesis. The recent discovery that allele III of papG(the P fimbrial adhesin gene) predominates among human cystitis isolates and confers an adherence phenotype resembling that of canine ExPEC prompted the present reevaluation of the canine-human ExPEC connection. Sixteen paired pap-positive urine and rectal E. coli isolates from dogs with urinary tract infection were studied. papG (adhesin) and papA(pilin) allele type, agglutination phenotypes, virulence factor genotypes, and randomly amplified polymorphic DNA and pulsed-field gel electrophoresis fingerprints were analyzed and compared with those of human ExPEC controls. The 16 canine strains contained predominantly papG allele III. Agglutination phenotypes segregated strictly according topapG allele status and were homogeneous among strains with the same papG allele profile irrespective of their human versus canine origin. Canine and human PapG variant III peptide sequences were highly homologous, without host species-specific differences. The most prevalent canine papA allele was F48, a novel variant recently identified among human urosepsis isolates. In addition to pap, human ExPEC-associated virulence genes detected among the canine strains included sfa/focDE,sfaS, fyuA, hlyA, cnf1,cdtB, kpsMT-II and -III, rfc,traT, ompT, and a marker for a pathogenicity-associated island from archetypal human ExPEC strain CFT073. Molecular fingerprinting confirmed the fecal origin of all but one canine urine isolate and showed one pair of O6 canine urine and fecal isolates to be extremely similar to an O6 human urosepsis isolate with which they shared all other genotypic and phenotypic characteristics analyzed. These data demonstrate that canine ExPEC strains are similar to, and in some instances essentially indistinguishable from, human ExPEC strains, which implicates dogs and their feces as potential reservoirs of E. coli with infectious potential for humans.

Identification of Urovirulence Traits in Escherichia coli by Comparison of Urinary and Rectal E. coli Isolates from Dogs with Urinary Tract Infection

ABSTRACT Spontaneously occurring urinary tract infection (UTI) in dogs was exploited as an experiment of nature to gain insights into UTI pathogenesis in humans. Concurrent urinary and rectal Escherichia coli isolates from 37 dogs with UTI were compared with respect to phylogenetic background, O antigens, and extended virulence genotype. In 54% of the UTI episodes, the dogs urinary and rectal isolates represented the same strain. Urinary isolates differed dramatically from rectal-only isolates in that they derived predominantly from E. coli phylogenetic group B2, expressed typical (human) UTI-associated O antigens, and possessed many virulence-associated genes, most notably pap elements (P fimbriae), papG (adhesin) allele III, sfa/foc and sfaS (S fimbriae), hly (hemolysin), fyuA (yersiniabactin), iroN (siderophore), and ompT (outer membrane protease T). The 20 urinary isolates that corresponded with the hosts predominant rectal strain were no less virulent according to the markers analyzed than were the 17 urinary isolates that differed from the hosts predominant rectal strain. These findings suggest that UTI pathogenesis is similar in dogs and humans, provide added support for the special-pathogenicity over the prevalence hypothesis of UTI pathogenesis, and identify numerous specific virulence-associated factors as significant correlates of urovirulence.

Recurrent and persistent urinary tract infections in dogs: 383 cases (1969-1995)

Laboratory records of bacterial urine cultures from 383 dogs with recurrent or persistent urinary tract infections (UTI) diagnosed at the University of California Veterinary Medical Teaching Hospital (VMTH) between 1969 and 1995 were reviewed retrospectively to characterize the bacteria involved and their association with age, gender, and breed of dogs affected. Sixty-eight breeds and a mixed-breed group were represented. Escherichia coli was the most common isolate, although mixed-bacterial infections were seen in 58% of the female and 55% of the male dogs. Recurrent and persistent UTI were most prevalent in middle-aged to older German shepherd dogs, miniature/toy poodles, and Labrador retrievers, with no apparent sex predilection. Criteria fitting recurrent and persistent UTI were present in 0.3% of all dogs seen at the VMTH during this 26-year period.

Mutations in the SLC2A9 Gene Cause Hyperuricosuria and Hyperuricemia in the Dog

Allantoin is the end product of purine catabolism in all mammals except humans, great apes, and one breed of dog, the Dalmatian. Humans and Dalmatian dogs produce uric acid during purine degradation, which leads to elevated levels of uric acid in blood and urine and can result in significant diseases in both species. The defect in Dalmatians results from inefficient transport of uric acid in both the liver and renal proximal tubules. Hyperuricosuria and hyperuricemia (huu) is a simple autosomal recessive trait for which all Dalmatian dogs are homozygous. Therefore, in order to map the locus, an interbreed backcross was used. Linkage mapping localized the huu trait to CFA03, which excluded the obvious urate transporter 1 gene, SLC22A12. Positional cloning placed the locus in a minimal interval of 2.5 Mb with a LOD score of 17.45. A critical interval of 333 kb containing only four genes was homozygous in all Dalmatians. Sequence and expression analyses of the SLC2A9 gene indicated three possible mutations, a missense mutation (G616T;C188F) and two promoter mutations that together appear to reduce the expression levels of one of the isoforms. The missense mutation is associated with hyperuricosuria in the Dalmatian, while the promoter SNPs occur in other unaffected breeds of dog. Verification of the causative nature of these changes was obtained when hyperuricosuric dogs from several other breeds were found to possess the same combination of mutations as found in the Dalmatian. The Dalmatian dog model of hyperuricosuria and hyperuricemia underscores the importance of SLC2A9 for uric acid transport in mammals.

Dried solidified blood calculi in the urinary tract of cats.

We have noted an increased number of calculi submitted to the Gerald V. Ling Urinary Stone Analysis Laboratory, University of California, Davis, that do not contain crystalline material but appear to be composed of dried solidified blood (DSB). Canine and feline laboratory records from 1986-2003 were reviewed for samples composed of >99% DSB. No calculi from dogs were found, but specimens from 49 cats were composed of >99% DSB, of which almost half (n = 22) had been submitted after 2001. The DSB calculi had been removed surgically or by postmortem examination from all areas of the upper and lower urinary tract. All samples were well formed and could be divided in half with Rongeur forceps. Detailed case information was available for 12 cats. Urinalyses were available for 9 of the 12 cats, and the mean specific gravity was 1.017 (range, 1.009-1.032). Red blood cells were reported in the sediment of all cats, with most containing >100 RBC/hpf. Ureteral obstructions but no radio-opaque calculi were visible by radiography, including radiographic contrast studies. Reports of ultrasound examinations were available for 10 cats, and discrete calculi were not recorded. In addition to polarized light microscopy, infrared spectroscopy and electron probe microanalysis were performed on a subset (n = 6, DSB calculi; n = 3, control calculi) of samples. Significantly more carbon, nitrogen, and sulfur (P = .012, P = .02, and P = .012, respectively) were present in the elemental analysis of the DSB calculi than in the control calculi, suggesting that the DSB calculi are primarily formed from organic material. At this time, we are uncertain why these DSB calculi become solidified, and we recommend that samples be submitted both in formalin and preservative free to further investigate their etiology.

Molecular investigation of Escherichia coli strains associated with apparently persistent urinary tract infection in dogs.

Persistent Escherichia coli urinary tract infection (UTI) in dogs is a frustrating clinical problem. Affected dogs often appear to fail to respond to therapy or to reacquire infection shortly after therapy is completed. Urovirulence factors (UVFs) of the infecting E. coli, antibiotic resistance, and tissue colonization may be contributory but have not been evaluated in dogs with persistent E. coli UTI. In this study, the strain types of E. coli in dogs with persistent UTI were evaluated with pulsed-field gel electrophoresis (PFGE) to determine whether persistence was due to acquisition of new isolates or failure to eradicate existing isolates. UVFs in these isolates, assessed by polymerase chain reaction, and antibiograms were correlated with treatment outcome in these dogs. Results documented a mixed pattern: 9 dogs remained chronically infected with 1 or 2 strains, each with distinct reproducible UVFs, but 1 dog was infected with numerous unrelated E. coli strains over time. Two dogs had a mixed pattern, consisting of 1 or more episodes of persistent E. coli infection attributable to a single strain in addition to episodes caused by unrelated strains. Many isolates had no detectable UVFs, highlighting the likely importance of impaired colonization resistance in the affected dogs. Antibiotic resistance was common, often in response to previous treatments, especially with trimethoprim-sulfamethoxazole. Antibiotic resistance patterns differed significantly within PFGE strain types, suggesting lateral acquisition of resistance plasmids or integrons. These results can be used to help guide testing for and management of persistent E. coli UTI in dogs.

Canine Uroliths: Analysis of Data Derived from 813 Specimens

This article contains an analysis of data compiled from 813 specimens of canine uroliths submitted to the Urinary Stone Analysis Laboratory at University of California School of Veterinary Medicine.

Methods of Analysis of Canine Uroliths

Accurate analysis of urinary calculi is essential for greater understanding of urinary stone disease and initiation of effective management and preventive measures. Quantitative and qualitative analysis of uroliths are discussed in this article.

Pancreatolithiasis and Pancreatic Pseudobladder Associated With Pancreatitis in a Cat

Pancreatolithiasis has been documented to occur naturally in humans and cattle. It has been associated with chronic pancreatitis in humans, and, when found, it may signify the presence of chronic pancreatic disease. This is the first report of a case involving a cat that had both an apparent obstruction with pancreatolithiasis as well as concurrent evidence of chronic pancreatic changes on histopathological evaluation. Additionally, this case documents the presence of a suspected congenital abnormality of a feline exocrine pancreas.

Bacterial culture of uroliths. Techniques and interpretation of results.

This article discusses techniques of bacterial culture of uroliths, interpretation of results of culture, and antimicrobial therapy of bacteria associated with uroliths.