f Molecular types of the Cryptococcus neoformans/Cryptococcus gattii species complex that infect dogs and cats differ regionally and with host species. Antifungal drug susceptibility can vary with molecular type, but the susceptibility of Cryptococcus isolates from dogs and cats is largely unknown. Cryptococcus isolates from 15 dogs and 27 cats were typed using URA5 restriction fragment length polymorphism analysis (RFLP), PCR fingerprinting, and multilocus sequence typing (MLST). Susceptibility was determined using a microdilution assay (Sensititre YeastOne; Trek Diagnostic Systems). MICs were compared among groups. The 42 isolates studied comprised molecular types VGI (7%), VGIIa (7%), VGIIb (5%), VGIIc (5%), VGIII (38%), VGIV (2%), VNI (33%), and VNII (2%), as determined by URA5 RFLP. The VGIV isolate was more closely related to VGIII according to MLST. All VGIII isolates were from cats. All sequence types identified from veterinary isolates clustered with isolates from humans. VGIII isolates showed considerable genetic diversity compared with other Cryptococcus molecular types and could be divided into two major subgroups. Compared with C. neoformans MICs, C. gattii MICs were lower for flucytosine, and VGIII MICs were lower for flucytosine and itraconazole. For all drugs except itraconazole, C. gattii isolates exhibited a wider range of MICs than C. neoformans. MICs varied with Cryptococcus species and molecular type in dogs and cats, and MICs of VGIII isolates were most variable and may reflect phylogenetic diversity in this group. Because sequence types of dogs and cats reflect those infecting humans, these observations may also have implications for treatment of human cryptococcosis.
Abstract. A case of fatal pulmonary hemorrhage in a 6-year-old American Paint mare with a 2-week history of intermittent coughing, fever, and epistaxis is described. Significant macroscopic abnormalities at postmortem examination were restricted to the respiratory system, and microscopically, severe pulmonary hemorrhage with suppurative bronchopneumonia was found. Actinobacillus equuli subsp. haemolyticus was cultured from a transtracheal wash performed antemortem as well as from the lungs at necropsy. The presence of airwayassociated hemorrhage in conjunction with bacterial bronchopneumonia suggested endothelial damage caused by a locally elaborated bacterial toxin, possibly produced by the A. equuli strain isolated from the lungs. The objective of this report was to indirectly document the presence of hemolysin repeat in structural toxin (RTX) in the lungs of the reported mare. A real-time polymerase chain reaction (PCR) assay targeting the recently described aqx gene of A. equuli subsp. haemolyticus was established and validated. Transcriptional activity of the aqx gene was used as a surrogate method to document toxin production. Real-time PCR analysis of the transtracheal fluid and lung tissue of the affected mare confirmed the presence and the transcriptional activity of the aqx gene at the genomic (gDNA) and complementary DNA (cDNA) levels, respectively. The presence of pneumonia associated with hemorrhagic pulmonary fluid and the culture of large numbers of hemolytic A. equuli should prompt the clinician to consider endothelial damage caused by bacterial toxins.
Three kittens, ages 5, 9, and 17 weeks, were found dead by separate caregivers and were submitted for necropsy. At gross necropsy, each kitten had hemorrhagic or bloody fibrinoserous thoracic fluid and differing distributions of pulmonary consolidation. On histologic examination, the pulmonary lesion in each kitten was similar and was characterized by acute necrotizing and hemorrhagic pneumonia and pleuritis, with numerous intralesional small Gram-negative rods. A pure culture of a distinct serotype of Escherichia coli was identified in lung tissue from each kitten (O4:H5, O6:H7, O6:H5). Lung isolates, genotyped by polymerase chain reaction, carried genes that are characteristic of extraintestinal pathogenic E. coli (ExPEC), including cnf-1, papG allele I, papA, papC, sfa, fim, hlyD, malX, iroN, fyuA, kpsMII, and ompT. Escherichia coli isolates from the intestines of 2 of the kittens were 100% related to the respective lung isolate, as determined by pulsedfield gel electrophoresis. Cultures of fecal samples collected from a clinically healthy cohort population of kittens revealed 16 of 19 tested kittens (84%) to be shedding hemolytic E. coli. Ten different serotypes were identified from 43 hemolytic E. coli fecal isolates from the cohort population, each of which had a genetic profile consistent with that typical of ExPEC. To the authors' knowledge, this is the first report to describe a cluster of isolated cases of pneumonia in kittens caused by distinct serotypes of ExPEC and to evaluate the prevalence of hemolytic E. coli carrying ExPEC-associated genes in the feces of a cohort population of kittens.
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