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Abstract

OBJECTIVE

To compare the bacteriome of the oral cavity in healthy dogs and dogs with various stages of periodontal disease.

ANIMALS

Dogs without periodontal disease (n = 12) or with mild (10), moderate (19), or severe (10) periodontal disease.

PROCEDURES

The maxillary arcade of each dog was sampled with a sterile swab, and swabs were submitted for next-generation DNA sequencing targeting the V1–V3 region of the 16S rRNA gene.

RESULTS

714 bacterial species from 177 families were identified. The 3 most frequently found bacterial species were Actinomyces sp (48/51 samples), Porphyromonas cangingivalis (47/51 samples), and a Campylobacter sp (48/51 samples). The most abundant species were P cangingivalis, Porphyromonas gulae, and an undefined Porphyromonas sp. Porphyromonas cangingivalis and Campylobacter sp were part of the core microbiome shared among the 4 groups, and P gulae, which was significantly enriched in dogs with severe periodontal disease, was part of the core microbiome shared between all groups except dogs without periodontal disease. Christensenellaceae sp, Bacteroidales sp, Family XIII sp, Methanobrevibacter oralis, Peptostreptococcus canis, and Tannerella sp formed a unique core microbiome in dogs with severe periodontal disease.

CONCLUSIONS AND CLINICAL RELEVANCE

Results highlighted that in dogs, potential pathogens can be common members of the oral cavity bacteriome in the absence of disease, and changes in the relative abundance of certain members of the bacteriome can be associated with severity of periodontal disease. Future studies may aim to determine whether these changes are the cause or result of periodontal disease or the host immune response.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To investigate the mycobiome of the oral cavity in healthy dogs and dogs with various stages of periodontal disease.

ANIMALS

51 dogs without periodontal disease (n = 12) or with mild (10), moderate (19), or severe (10) periodontal disease.

PROCEDURES

The whole maxillary arcade of each dog was sampled with a sterile swab, and swabs were submitted for next-generation DNA sequencing targeting the internal transcribed spacer 2 region with a commercial sequencing platform.

RESULTS

Fungi were detected in all samples, with a total of 320 fungal species from 135 families detected in the data set. No single fungal species was found in all samples. The 3 most frequently found fungal species were Cladosporium sp (46/51 samples), Malassezia restricta (44/51 samples), and Malassezia arunalokei (36/51 samples). Certain fungi, specifically those of the family Didymellaceae, the family Irpicaceae, and the order Pleosporales, were significantly associated with different stages of periodontitis. Mycobial analysis indicated that Cladosporium sp could be considered part of the core oral cavity mycobiome.

CONCLUSIONS AND CLINICAL RELEVANCE

Results highlighted that fungi are present in the oral cavity of dogs and are characterized by substantial species diversity, with different fungal communities associated with various stages of periodontal disease. The next-generation DNA sequencing used in the present study revealed substantially more species of fungi than previous culture-based studies.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To determine whether a stainless steel implant sterilized with a novel cold atmospheric plasma sterilization (CAPS) device adversely affects local tissues in rabbits and whether CAPS was as effective as steam sterilization with an autoclave to inactivate Pasteurella multocida.

ANIMALS

31 healthy New Zealand White rabbits.

PROCEDURES

Steam-autoclaved stainless steel implants inoculated with P multocida underwent a second steam autoclave sterilization (AIA) or CAPS (AICAPS). One AIA implant and 3 AICAPS implants were randomly placed subcutaneously at 4 sites in 21 rabbits (84 implants). These rabbits were monitored daily for 5 days for evidence of systemic illness and local tissue reactions at the implantation sites and then euthanized. Samples were taken from each implant site for bacterial culture and histologic examination.

RESULTS

Cultures of samples obtained from all sites were negative for bacterial growth. No significant difference was observed in mean skin thickness or erythema between AIA and AICAPS implant sites on any observed day. Also, individual histologic grades for the epidermis, dermis, subcutis, and muscle and total histologic grade were not significantly different between AIA and AICAPS implant sites.

CONCLUSIONS AND CLINICAL RELEVANCE

Cold atmospheric plasma sterilization was noninferior to steam sterilization of P multocida–contaminated stainless steel implants in the rabbits in the present study. However, studies of the efficacy of CAPS for inactivation of other important bacteria are needed.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To evaluate erythema and number of CFUs on the skin of dogs with hair clipped by use of 2 sizes of clipper blades.

ANIMALS

67 client-owned dogs receiving an epidural.

PROCEDURES

Hair was clipped with a No. 10 blade (approx hair length, 1.5 mm) on one half and a No. 40 blade (approx hair length, 0.25 mm) on the other half of each epidural site. Skin was surgically scrubbed with 2% chlorhexidine gluconate and 70% isopropyl alcohol. Samples were obtained immediately after clipping, after skin was scrubbed, and again 24 hours after clipping. Number of CFUs for both sides of the clipped areas, types of microorganisms, and growth on MacConkey agar were evaluated every 24 hours for 72 hours. Colonies were evaluated for bacterial morphology and Gram stain characteristics. Sites were evaluated 24 hours after clipping for evidence of erythema.

RESULTS

24 hours after hair was clipped, there was a significantly higher incidence of erythema and higher number of Micrococcaceae bacteria for the side clipped with the No. 40 blade than the side clipped with the No. 10 blade. Number of CFUs did not differ significantly between size of clipper blades.

CONCLUSIONS AND CLINICAL RELEVANCE

Clipping hair with a No. 40 blade resulted in a significant increase in the incidence of erythema and higher number of Micrococcaceae bacteria, compared with results for clipping with a No. 10 blade. These results supported use of a No. 10 clipper blade to prevent erythema and reduce variation in the skin microbiome.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To evaluate a fluorescence resonance energy transfer quantitative PCR (FRET-qPCR) assay for detection of gyrA mutations conferring fluoroquinolone resistance in canine urinary Escherichia coli isolates and canine urine specimens.

SAMPLE 264 canine urinary E coli isolates and 283 clinical canine urine specimens.

PROCEDURES The E coli isolates were used to validate the FRET-qPCR assay. Urine specimens were evaluated by bacterial culture and identification, isolate enrofloxacin susceptibility testing, and FRET-qPCR assay. Sensitivity and specificity of the FRET-qPCR assay for detection of gyrA mutations in urine specimens and in E coli isolated from urine specimens were computed, with results of enrofloxacin susceptibility testing used as the reference standard.

RESULTS The validated FRET-qPCR assay discriminated between enrofloxacin-resistant and enrofloxacin-susceptible E coli isolates with an area under the receiver operating characteristic curve of 0.92. The assay accurately identified 25 of 40 urine specimens as containing enrofloxacin-resistant isolates (sensitivity, 62.5%) and 226 of 243 urine specimens as containing enrofloxacin-susceptible isolates (specificity, 93.0%). When the same assay was performed on E coli isolates recovered from these specimens, sensitivity (77.8%) and specificity (94.8%) increased. Moderate agreement was achieved between results of the FRET-qPCR assay and enrofloxacin susceptibility testing for E coli isolates recovered from urine specimens.

CONCLUSIONS AND CLINICAL RELEVANCE The FRET-qPCR assay was able to rapidly distinguish between enrofloxacin-resistant and enrofloxacin-susceptible E coli in canine clinical urine specimens through detection of gyrA mutations. Therefore, the assay may be useful in clinical settings to screen such specimens for enrofloxacin-resistant E coli to avoid inappropriate use of enrofloxacin and contributing to antimicrobial resistance.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To evaluate the correlation between the density of native gastric Helicobacter spp and the presence of gastric lesions in dogs.

ANIMALS 80 dogs of various breeds, sexes, and ages.

PROCEDURES Gastroscopic and histologic examinations were performed for all dogs. Helicobacter spp were detected by combining evaluation of urease activity and results of bacteriologic culture, microscopic observation, and a 16S rRNA PCR assay. The density of Helicobacter-like organisms was evaluated with light microscopy by use of Warthin-Starry modified stain. Correlations were evaluated by use of the Spearman correlation analysis.

RESULTS Gastritis was found in 55 of 80 dogs and classified as mild (n = 31), moderate (16), or severe (8). Of these 55 dogs, only 8 had clinical signs. Histologic examination revealed some degree of lymphocytic-plasmacytic infiltrate, mild eosinophilia, and neutrophilic inflammation in the lamina propria. Seventy-six dogs had positive results for Helicobacter spp. Helicobacter pylori DNA was not detected. Low density and homogeneous distribution of Helicobacter spp were observed in all gastric zones.

CONCLUSIONS AND CLINICAL RELEVANCE A significant correlation between density of Helicobacter spp and gastroscopic or histologic lesions was not detected. These findings supported the contention that there is no correlation between general Helicobacter spp density or numbers and gastritis in dogs.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To evaluate a hypervariable octameric oligonucleotide fingerprints (HOOF-Prints) assay for identification of and discrimination between wild-type and vaccine strains of Brucella melitensis.

SAMPLE Brucella melitensis vaccine strain M5 and wild-type strain M43.

PROCEDURES 8 pairs of primers (alterable, octameric nucleotides) were designed on the basis of a biological analysis of 8 flanking sequences in the DNA of B melitensis. The HOOF-Prints technique was used to identify wild-type and vaccine strains of B melitensis. Phylogenetic analysis of short, polymorphic fragments of DNA from B melitensis strains M5 and M43 was performed.

RESULTS Variable-number tandem repeat DNA segments of B melitensis vaccine strain M5 and wild-type strain M43 were successfully amplified by means of PCR assay. All target gene fragments ranged in size from 100 to 300 bp. Separate phylogenetic analysis of each Brucella strain revealed considerable differences between the vaccine and wild-type strains.

CONCLUSIONS AND CLINICAL RELEVANCE The results of this study suggested the HOOF-Prints assay may be useful for discriminating vaccine strains of B melitensis from wild-type strains. This ability could allow discrimination between animals that are seropositive because of vaccination against B melitensis and those that are seropositive because of B melitensis infection and could decrease the likelihood of importing Brucella-infected animals.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To determine whether consumption of a single dental treat with specific mechanical properties and active ingredients would provide a 24-hour effect on dental plaque bacteria and halitosis in dogs.

ANIMALS 10 dogs of various breeds from a privately owned colony that had received routine dental scaling and polishing 4 weeks before the study began.

PROCEDURES Dogs were randomly assigned to receive 1 placebo or dental treat first. A 4-week washout period was provided, and then dogs received the opposite treatment. Oral plaque and breath samples were collected before and 0.5, 3, 12, and 24 hours after treat consumption. Volatile sulfur compounds (VSCs) concentration was measured in breath samples. Total aerobic, total anaerobic, Porphyromonas gulae, Prevotella intermedia–like, Tannerella forsythia, and Fusobacterium nucleatum bacterial counts (measured via bacterial culture) and total live bacterial counts, total live and dead bacterial counts, and bacterial vitality (measured via quantitative real-time PCR assay) were assessed in plaque samples.

RESULTS Compared with placebo treat consumption, dental treat consumption resulted in a significant decrease in breath VSCs concentration and all plaque bacterial counts, without an effect on bacterial vitality. Effects of the dental treat versus the placebo treat persisted for 12 hours for several bacterial counts and for 24 hours for breath VSCs concentration.

CONCLUSIONS AND CLINICAL RELEVANCE Although clinical benefits should be investigated in larger scale, longer-term studies, results of this study suggested that feeding the evaluated dental treat may help to decrease oral bacterial growth in dogs for 12 hours and oral malodor for 24 hours. A feeding interval of 12 hours is therefore recommended.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To determine effects of in ovo administration of a probiotic on development of the intestinal microbiota of 2 genetic lineages (modern and heritage) of chickens.

SAMPLE 10 newly hatched chicks and 40 fertile eggs to determine intestinal microbiota at hatch, 900 fertile eggs to determine effects of probiotic on hatchability, and 1,560 chicks from treated or control eggs.

PROCEDURES A probiotic competitive-exclusion product derived from adult microbiota was administered in ovo to fertile eggs of both genetic lineages. Cecal contents and tissues were collected from embryos, newly hatched chicks, and chicks. A PCR assay was used to detect bacteria present within the cecum of newly hatched chicks. Fluorescence in situ hybridization and vitality staining were used to detect viable bacteria within intestines of embryos. The intestinal microbiota was assessed by use of 16S pyrosequencing.

RESULTS Microscopic evaluation of embryonic cecal contents and tissues subjected to differential staining techniques revealed viable bacteria in low numbers. Development of the intestinal microbiota of broiler chicks of both genetic lineages was enhanced by in ovo administration of adult microbiota. Although the treatment increased diversity and affected composition of the microbiota of chicks, most bacterial species present in the probiotic were transient colonizers. However, the treatment decreased the abundance of undesirable bacterial species within heritage lineage chicks.

CONCLUSIONS AND CLINICAL RELEVANCE In ovo inoculation of a probiotic competitive-exclusion product derived from adult microbiota may be a viable method of managing development of the microbiota and reducing the prevalence of pathogenic bacteria in chickens.

Restricted access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To determine the diagnostic accuracy of a rapid immunoassay (RIA) for point-of-care detection of urinary tract infection (UTI) of dogs, compared with criterion-referenced diagnosis with bacterial culture.

SAMPLE 200 urine samples obtained from dogs and submitted to a veterinary microbiology diagnostic laboratory for routine bacterial culture and antimicrobial susceptibility determination.

PROCEDURES Samples were evaluated by use of quantitative bacterial culture and the RIA. Sensitivity, specificity, and positive and negative predictive values of the RIA were calculated; results of bacterial culture were the criterion-referenced outcome. A κ statistic was calculated to determine agreement between bacterial culture and RIA results.

RESULTS 56 of 200 (28%) urine samples had positive results for bacterial growth by use of culture methods; there were 38 (19%) positive results likely to be associated with bacterial UTI on the basis of sample collection method and bacterial concentration. Sensitivity and specificity of the RIA for detecting samples likely to be associated with UTI (≥ 1,000 CFUs/mL) were 97.4% and 98.8%, respectively. The positive and negative predictive values of the RIA for bacterial cultures with likely UTI were 0.949 and 0.994, respectively. Agreement between bacterial culture and RIA outcome for UTI was substantial (weighted κ, 0.718).

CONCLUSIONS AND CLINICAL RELEVANCE The RIA test evaluated in this study accurately detected UTI of dogs, compared with detection with the criterion-referenced bacterial culture method. Use of this point-of-care RIA could allow clinicians to diagnose UTI at the time of a patient visit and provide information useful for immediately initiating empirical antimicrobial treatment. (Am J Vet Res 2016;77:162–166)

Restricted access
in American Journal of Veterinary Research