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Abstract

Objective—To determine the prevalence of 4 urovirulence genes in fecal Escherichia coli isolates from healthy dogs and their owners and to determine whether detection of E coli strains with these genes was associated with a history of urinary tract infection (UTI).

Sample Population—61 healthy dog-owner pairs and 30 healthy non–dog owners.

Procedures—A fecal specimen was obtained from each participant, and 3 colonies of E coli were isolated from each specimen. A multiplex PCR assay was used to detect 4 genes encoding virulence factors: cytotoxic necrotizing factor (cnf), hemolysin (hlyD), s-fimbrial and F1C fimbriae adhesin (sfa/foc), and pilus associated with pyelonephritis G allele III (papGIII). Human participants completed a questionnaire to provide general information and any history of UTI for themselves and, when applicable, their dog.

Results—26% (16/61) of dogs, 18% (11/61) of owners, and 20% (6/30) of non–dog owners had positive test results for ≥ 1 E coli virulence gene. One or more genes were identified in fecal E coli isolates of both dog and owner in 2% (1/61) of households. There was no difference in the detection of any virulence factor between dog-owner pairs. Female owner history of UTI was associated with detection of each virulence factor in E coli strains isolated from their dogs' feces.

Conclusions and Clinical Relevance—Dogs and humans harbored fecal E coli strains possessing the genes cnf, hlyD, sfa/foc, and papGIII that encode urovirulence factors. It was rare for both dog and owner to have fecal E coli strains with these virulence genes.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To estimate the relationship between therapeutic use of ceftiofur and recovery of Escherichia coli and Salmonella spp with reduced susceptibility to ceftriaxone from feces of dairy cattle.

Animals—3,840 mature dairy cows on 50 dairy herds in Ohio.

Procedures—Fecal samples were obtained from up to 100 mature dairy cows on each farm. Samples were screened for E coli and Salmonella spp with reduced susceptibility to ceftriaxone by use of selective media.

ResultsE coli with reduced susceptibility to ceftriaxone was recovered from 92% (46/50) of the herds and 60.9% (2,338/3,840) of cows. Salmonella spp were recovered from 44% (22/50) of the herds and 9.9% (382/3,840) of cows. No association was found between ceftiofur use and recovery of E coli with reduced susceptibility to ceftriaxone at the herd level. However, recovery of E coli with reduced susceptibility to ceftriaxone was more likely from cows in herds in which Salmonella spp were also recovered on the day of collection (odds ratio, 24.96; 95% confidence interval, 3.17 to 196.68) than from herds in which Salmonella spp were not recovered. Odds of recovery of E coli with reduced susceptibility to ceftriaxone from an individual cow increased 62% (odds ratio, 1.62; 95% confidence interval, 1.16 to 2.25) for every 454-kg increase in herd milk production.

Conclusions and Clinical Relevance—No evidence was found that the use of ceftiofur on dairy farms increases the prevalence or dissemination of Salmonella spp or E coli with reduced susceptibility to ceftriaxone.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To evaluate the radial growth assay for use in in vitro susceptibility testing of Pythium insidiosum and a Lagenidium sp and to assess susceptibility of representative isolates to itraconazole, posaconazole, voriconazole, terbinafine, caspofungin, and mefenoxam.

Sample Population—6 isolates each of P insidiosum and Lagenidium sp.

Procedures—Isolates were plated in triplicate onto agar supplemented with antifungal compounds at concentrations of 0.025 to 8 μg/mL. Isolates on dimethyl sulfoxide– and water-supplemented agar served as control samples. Effect of antifungal concentration on colony diameter was assessed with a mixed linear model. Assay variability was assessed with the coefficient of variation.

Results—Colony growth was uniform (mean intra-assay and interassay coefficients of variation were < 5%). Minimal inhibition was evident with voriconazole and posaconazole at 8 μg/mL. Terbinafine at 8 μg/mL significantly reduced growth of P insidiosum and at ≥ 1 μg/mL significantly reduced growth of the Lagenidium sp. Caspofungin and mefenoxam (concentrations ≥ 1 μg/mL and ≥ 0.025 μg/mL, respectively) significantly reduced growth of both pathogens. Mefenoxam at 0.1 μg/mL caused > 50% growth inhibition in 11 of 12 isolates and at 1 μg/mL caused > 90% inhibition in all isolates.

Conclusions and Clinical Relevance—Results suggested that the radial growth assay was a simple, reproducible technique for susceptibility testing of P insidiosum and a Lagenidium sp. Azoles had limited activity, whereas terbinafine and caspofungin caused significant but minimal to moderate inhibition. Only mefenoxam had a profound effect on both pathogens at concentrations likely to be achievable in tissues.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine whether groups C and G streptococci (GCS-GGS) isolated from animals have rheumatogenic traits associated with human GCS-GGS isolates, particularly the potential of the bacteria to interact with human collagen type IV (collagen-IV), known to be targeted during acute rheumatic fever (ARF).

Sample Population—64 GCS and GGS bacterial strains isolated from infected animals.

Procedures—Bacteria were analyzed for their ability to bind and aggregate collagen-IV and for the presence of collagen binding factors, such as the hyaluronic acid capsule, cne gene, and emm gene.

Results—Collagen-IV binding ability was detected in 19% (n = 12) of the isolates studied. Of the collagen-IV binding strains, 5 expressed hyaluronic acid capsule. Furthermore, emm was detected in the genome of 1 isolate, whereas all remaining collagen-IV binding isolates possessed the cne gene. Of the collagen binding factors investigated, the hyaluronic capsule was the only factor for which collagen-IV interaction could be detected. Investigation of the potential of these strains to aggregate collagen-IV revealed that animal isolates had a nonaggregating phenotype.

Conclusions and Clinical Relevance—Despite efficiently binding collagen-IV via hyaluronic acid, animal isolates lacked the ability to initiate aggregation of this protein. Because collagen-IV aggregation is associated with all collagen-IV–binding rheumatogenic strains, this suggested a lack of rheumatogenic potential among animal-derived GCS and GGS and, therefore, a low chance of acquiring ARF through animal contact.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine whether the active metabolite of leflunomide, A77 1726 (A77), inhibits replication of feline herpesvirus-1 (FHV-1) in cell culture.

Study Population—Crandell Rees feline kidney (CRFK) cell cultures.

Procedures—Cell cultures were inoculated with FHV-1 and treated simultaneously with concentrations of A77 ranging from 0 to 200μM. The antiviral effect of A77 was determined by use of conventional plaque reduction assays. The effect of A77 on viral load was determined via real-time PCR analysis, and transmission electron microscopy was used to evaluate the effect of A77 on viral morphology. To determine whether the antiviral effect was attributable to alterations in CRFK cell viability and number, CRFK cells were treated with various concentrations of A77 and stained with Annexin V and propidium iodide to assess apoptosis and a mitochondrial function assay was used to determine cell viability.

Results—Concentrations of A77 ≥ 20μM were associated with substantial reduction in plaque number and viral load. Concentrations ≥ 100μM were associated with complete suppression of plaque formation. At low concentrations of A77, clusters of intracytoplasmic virus particles that appeared to lack tegument and an external membrane were detected. Treatment of uninfected CRFK cell monolayers with A77 was associated with reduction in mitochondrial function with minimal evidence of apoptosis.

Conclusions and Clinical Relevance—Leflunomide may be an alternative to current calcineurin-based immunosuppressive protocols used in feline organ transplantation because of its antiherpesviral activity.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine the effect of nonthermal plasma on Staphylococcus aureus, fibroblasts in monolayer culture, and clean and contaminated skin explants.

Sample Population—Normal skin from euthanized horses.

ProceduresS aureus organisms were plated and treated with nonthermal plasma followed by bacterial culture to assess viability. Fibroblasts in monolayer culture and the epidermal and dermal surfaces of clean and S aureus–contaminated skin explants were treated. The effects of distance and duration on the response to treatment were compared.

Results—Compared with controls, treatment with nonthermal plasma resulted in significantly decreased bacterial growth and significantly inhibited survival of fibroblasts in monolayer culture. When epidermal and dermal surfaces of skin explants were treated, there was no effect on production of normal fibroblasts during explant culture, except when extended exposure times of ≥ 2 minutes were used. Treatment with nonthermal plasma resulted in significantly lower bacterial counts after 24 hours of culture of S aureus–contaminated epidermis but not of dermis.

Conclusions and Clinical Relevance—Nonthermal plasma resulted in bacterial decontamination of agar and epithelium; negative effects on fibroblasts in monolayer; and no negative effects on skin explants, except at long exposure times. Use of nonthermal plasma appears safe for treatment of epithelialized surfaces, may be safe for granulating wounds, and results in decontamination of S aureus. Investigations on the effects that nonthermal plasma may have on patient tissues are indicated with a clinically applicable delivery device.

Full access
in American Journal of Veterinary Research

Abstract

Objective—To compare clinical information obtained from medical records of cats with methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S aureus (MSSA) infections, evaluate antibiograms of MRSA and MSSA for multiple-drug resistance (MDR), and characterize the strain type and staphylococcal chromosome cassette (SCC)mec type of each MRSA.

Sample Population—70 S aureus isolates obtained from 46 cats.

Procedures—Clinical information obtained from medical records, including signalment, clinical signs, histologic examination of affected tissues, and outcomes, was compared between the 2 groups. Composite antibiograms of MRSA and MSSA were compared statistically. The MRSA strains were characterized by use of pulsed-field gel electrophoresis and SCCmec typing.

Results—No statistical differences in signalment or subjective differences in clinical signs or outcomes were detected between groups with MRSA or MSSA infection. Significant differences in antimicrobial resistance were detected, with MRSA having complete resistance to fluoroquinolone and macrolide antimicrobials, whereas MSSA maintained a high frequency of susceptibility. Seven pulsed-field patterns were observed in 15 MRSA strains; all but 1 were highly related. All MRSA isolates contained a type II SCCmec element.

Conclusions and Clinical Relevance—Because MDR cannot be predicted in staphylococcal infections in cats on the basis of clinical signalment, culture and susceptibility testing are recommended whenever initial empirical treatment is unsuccessful. Molecular characterization of MRSA strains suggests that there has been reverse-zoonotic transmission from humans.

Impact for Human Medicine—The SCCmec type II element is typically associated with nosocomial MRSA infections of people. Cats may serve as reservoirs for MRSA infections in humans.

Full access
in American Journal of Veterinary Research

Abstract

Objective

To characterize 2 strains of Haemobartonella felis by use of molecular techniques.

Animals

35 specific-pathogen-free cats, 6 months to 4 years old.

Procedure

Intraperitoneal or IV inoculation with blood containing H felis small form (Hfsm, 18 cats) or H felis large form (Hflg, 11 cats); 6 cats were uninfected controls. Hfsm was evaluated for capability to cross-protect against the more virulent Hflg. Morphology of both strains was compared by light microscopy of Wright-Giemsa-stained blood smears, and the 16S rRNA genes were sequenced.

Results

Infection with Hflg induced signs of depression, fever, and severe macrocytic normochromic anemia with nucleated erythrocytes. More than 95% of erythrocytes were parasitized. Inoculation with Hfsm and uninfected control blood induced mild or no clinical signs and no hematologic abnormalities. Anti-Hfelis lgG was first detected on postinoculation day (PID) 21, and increased to maximal titer of 400 by PID 28. Reactivated infection was observed in 8 of 29 cats (4 Hfsm and 4 Hflg), with 5% parasitized erythrocytes during the later attack. On PID 8, Hflg-inoculated cats had positive results of polymerase chain reaction analysis (PCR) that persisted until cats were treated with doxycycline or oxytetracycline; Hfsm-inoculated cats had positive PCR results that persisted for duration of observation (3 months).

Conclusions

Genetically and morphologically distinct strains of H felis infect cats in the field. The level of genetic difference suggested that these strains may be different species or genera.

Clinical Relevance

PCR is a critical diagnostic aid to detect occult Haemobartonella spp infection, as well as response to treatment and clearance of the organism. (Am J Vet Res 1998;59:1581-1588)

Free access
in American Journal of Veterinary Research

Abstract

Objectives

To determine efficacy of orally administered Brucella abortus vaccine strain RB51 against virulent B abortus challenge exposure in cattle as a model for vaccination of wild ungulates.

Animals

20 mixed-breed beef cattle obtained from a brucellosis-free herd.

Procedure

Sexually mature, Brucella-negative beef heifers were vaccinated by mixing > 1010 viable RB51 organisms or diluent with their feed. Heifers were fed individually and consumed their entire ration. Each heifer received approximately 3×1010 colony-forming units (CFU). Six weeks after oral vaccination, heifers were pasture-bred to brucellosis-free bulls. At approximately 186 days' gestation, heifers were challenge exposed conjunctively with 107 CFU of virulent B abortus strain 2308.

Results

Vaccination with the rough variant of B abortus RB51 did not stimulate antibodies against the O-polysaccharide (OPS) of B abortus. After challenge exposure and parturition, strain 2308 was recovered from 80% of controls and only 20% of vaccinates. Only 30% of the vaccinates delivered dead, premature, or weak calves, whereas 70% of the controls had dead or weak calves.

Conclusions

Cattle vaccinated orally with the rough variant of B abortus strain RB51 develop significant (P< 0.05) protection against abortion and colonization and do not produce OPS-specific antibodies.

Clinical Relevance

Results encourage further investigation into use of strain RB51 to vaccinate wild ungulates (elk and bison) orally. (Am J Vet Res1998; 59:1575-1578)Vol 59, No. 12, December 1998

Free access
in American Journal of Veterinary Research

Abstract

Objective

To determine the prevalence of antibody to bovine adenovirus (BAdV) serotypes 1-8 and 10 in calves at a farm and after 5 weeks in a feedyard.

Animals

2- to 5-month-old calves of mixed English breeding (n = 100) from 4 farms.

Procedure

Serum BAdV antibody was measured by use of a microtitration test.

Results

Serum antibodies were found to the 9 BAdV serotypes studied. Seroconversion to each virus had occurred in some calves by the time the second serum sample had been obtained, indicating that the BAdV were present and inducing active infection in these calves.

Conclusions

Antibody to BAdV serotypes 1-8 and 10 are present in cattle populations of the United States, indicating existence of these serotypes, although only BAdV serotypes 1-4, 7, and 10 have been isolated. (Am J Vet Res 1998;59:1579-1580)

Free access
in American Journal of Veterinary Research