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An organism reported in the early literature to be a rare cause of cervical lymphadenitis in guinea pigs, Streptobacillus moniliformis, has been reclassified as Caviibacter abscessus We describe a series of sequential cases of abscesses in guinea pigs that were presented to our clinic from which the only agent isolated was a unique, serum-requiring bacterium. Discrete colonies were not detected in 6.5% CO 2 or anaerobically on routine primary isolation media containing up to 5% whole sheep blood, with and without cysteine, vitamin K, and hemin supplementation after 7 days of incubation at 37°C. Based on subsequently determined growth requirements, the organisms were best described as serum-requiring, aerotolerant anaerobes. Colonies were detectable within 24 h at 37°C in an anaerobic atmosphere on a mycoplasma agar-based medium containing 10% pig serum and reached 3 mm in diameter within 3-5 days. Microscopic appearance consisted of small gram-negative rods and coccobacilli with occasional filaments. However, in direct smears from clinical specimens and from weak or dysgonic growth on plates incubated under suboptimal growth conditions (e.g., in 6.5% CO 2 ), irregular rods with occasional small bulbous forms or numerous long wavy filaments were observed. All of the isolates generated unique spectral profiles similar to that of C. abscessus when examined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Phylogenetic analysis of partial 16S rRNA gene sequences showed that the isolates were identical to each other and shared 99.9% sequence identity with C. abscessus., (© 2016 The Author(s).)

Feline immunodeficiency virus (FIV) is an important infectious agent of cats. Clinical syndromes resulting from FIV infection include immunodeficiency, opportunistic infections, and neoplasia. In our study, a 5' long terminal repeat/gag region-based reverse transcription insulated isothermal polymerase chain reaction (RT-iiPCR) was developed to amplify all known FIV strains to facilitate point-of-need FIV diagnosis. The RT-iiPCR method was applied in a point-of-need PCR detection platform--a field-deployable device capable of generating automatically interpreted RT-iiPCR results from nucleic acids within 1 hr. Limit of detection 95% of FIV RT-iiPCR was calculated to be 95 copies standard in vitro transcription RNA per reaction. Endpoint dilution studies with serial dilutions of an ATCC FIV type strain showed that the sensitivity of lyophilized FIV RT-iiPCR reagent was comparable to that of a reference nested PCR. The established reaction did not amplify any nontargeted feline pathogens, including Felid herpesvirus 1, feline coronavirus, Feline calicivirus, Feline leukemia virus, Mycoplasma haemofelis, and Chlamydophila felis. Based on analysis of 76 clinical samples (including blood and bone marrow) with the FIV RT-iiPCR, test sensitivity was 97.78% (44/45), specificity was 100.00% (31/31), and agreement was 98.65% (75/76), determined against a reference nested-PCR assay. A kappa value of 0.97 indicated excellent correlation between these 2 methods. The lyophilized FIV RT-iiPCR reagent, deployed on a user-friendly portable device, has potential utility for rapid and easy point-of-need detection of FIV in cats., (© 2015 The Author(s).)

Nonhemolytic isolates of catalase-positive, Gram-positive cocci were observed that produced strong synergistic hemolysis with colonies of Staphylococcus pseudintermedius and Staphylococcus schleiferi subsp. coagulans in primary cultures of clinical specimens from dogs. The isolates appeared similar to nonhemolytic staphylococci but were not identified with conventional biochemical tests or commercial biochemical test kits. Partial 16S ribosomal RNA gene sequences from each of 9 tested isolates were nearly identical to that of Rothia nasimurium. The isolates produced positive Christie-Atkins-Munch-Peterson test reactions with Staphylococcus aureus (ATCC strain 25923) and Staphylococcus intermedius Hajek (ATCC strain 49052). The latter was identified in the current study as S. pseudintermedius. Rothia nasimurium appears to be a previously unreported commensal organism in dogs. Its potential significance as an opportunistic contributor to polymicrobial infections in dogs is not known., (© 2014 The Author(s).)

Squamous cell carcinoma (SCC) is the most common tumor arising in the periocular and penile areas of horses. Both ultraviolet radiation and papillomaviruses have been implicated in the pathogenesis of SCC in various species, including the horse. This retrospective study used polymerase chain reaction (PCR) to detect papillomavirus DNA in archival biopsy samples from equine periocular and penile SCC from 3 different geographic areas (northeast, southeast, and central United States). Forty-two periocular SCCs were tested; none contained papillomavirus DNA. Twenty-two penile SCCs were tested, and papillomavirus DNA was identified in 10 (43%) cases. Sequencing of the PCR products revealed homology with Equus caballus papillomavirus 2 (EcPV-2). No geographic distribution in the detection of papillomavirus was identified. Penile SCCs were significantly more likely to be papillomavirus positive than the periocular SCCs (P < 0.001). The role of papillomavirus in the development of penile SCC requires further investigation. The differing pathogeneses of periocular and penile SCC suggest that the tumors may respond differently to treatment.

Cefoxitin disk diffusion susceptibility testing is a recommended screening method for the detection of methicillin resistance in human isolates of Staphylococcus aureus and coagulase-negative staphylococci. A retrospective analysis of 1,146 clinical isolates of Staphylococcus pseudintermedius from dogs was conducted to determine if screening by the cefoxitin disk method can be similarly useful with S. pseudintermedius. The distribution of cefoxitin growth inhibition zone diameters within this collection was bimodal and correlated well with the results of methicillin resistance gene (mecA) detection by polymerase chain reaction. Of the isolates, 5% had discordant results and, when retested, 84% of these were in agreement. While a greater diversity of isolates and interlaboratory comparisons must be tested, the current study suggests that an epidemiological breakpoint (of approximately ≤ 30 mm = resistant; ≥ 31 = susceptible) can be established to predict methicillin resistance in S. pseudintermedius.

The present case report describes isolation of the fungus Oxyporus corticola from multiple lymphocutaneous tissues of a Beagle dog. Until recently, this fungus had not been reported in the human or veterinary medical literature as a cause of animal disease. A single previous report also involved infection in a German Shepherd Dog, a breed with reported increased susceptibility to disseminated fungal infection and dysfunctional immune response. Isolates were non-sporulating and required molecular identification methods for prompt differentiation from other fungal pathogens. Risk factors for infection with O. corticola are unknown.

Isolates of gram-negative anaerobic bacteria from reptiles have only occasionally been identified to the genus and species level in the veterinary medical literature. In particular, reports identifying Porphyromonas spp. from infections in reptiles are scarce. The present report describes unique Porphyromonas isolates obtained from necrosuppurative infections in central bearded dragons (Pogona vitticeps). The isolates grew in the presence of oxygen, were strongly hemolytic, and did not produce detectable black, iron porphyrin pigment. Biochemical identification kit numeric biocodes gave high but unreliable probabilities (>99.9%) for identification as Porphyromonas gingivalis. Partial 16S ribosomal RNA gene sequences of the isolates were identical to each other and shared 91% identity with those of Porphyromonas gulae. The isolates may represent a new reptile-associated Porphyromonas species.

Lycoperdonosis is a rare respiratory disease that results from the inhalation of spores released from the Lycoperdon (puffball) mushroom. In the present study, 2 cases of confirmed canine lycoperdonosis are described. The first case presented to the Matthew J. Ryan Veterinary Hospital of the University of Pennsylvania, and the second case was submitted for postmortem examination to the University of Tennessee Veterinary Teaching Hospital. Both dogs presented in respiratory distress, and owners reported that the dogs had been playing or digging in areas with puffball mushrooms prior to the onset of clinical signs. In the initial case, thoracic radiographs revealed a diffuse interstitial and multifocal alveolar pulmonary pattern. Despite aggressive medical treatment and mechanical ventilation, the dog continued to worsen and was euthanized. Postmortem examination revealed firm lung lobes and enlarged tracheobronchial lymph nodes. Histologically, there was a severe diffuse histiocytic and pyogranulomatous bronchointerstitial pneumonia. Throughout the lung and lymph nodes, most commonly within macrophages, were round, 3-5 µm in diameter, Gomori methenamine silver-positive structures, consistent with Lycoperdon spores. An approximately 750-base pair DNA fragment was amplified from lung of both cases by polymerase chain reaction using primers specific to yeast ribosomal DNA, and the sequence of the fragment was determined to be most closely related to Lycoperdon pyriforme. Importantly, reexamination of an endotracheal wash from the initial case revealed intrahistiocytic spores, suggesting that airway sampling may assist in diagnosing lycoperdonosis.

On the basis of the scarcity of reports in the veterinary literature, it appears that Propionibacterium spp. are rarely associated with disease or isolated from cattle tissues. Recently, Propionibacterium spp. has been associated with multifocal abscessation in cattle. This report describes a case of necrosuppurative placentitis and abortion in an adult Holstein cow. Numerous colonies of small, pleomorphic, Gram-positive, rod-shaped bacteria were observed within the fibrin lattice associated with placental lesions and within the fetal atelectatic lung. Propionibacterium acnes was isolated in high numbers from the placenta, fetal lung, and stomach contents. To the authors' knowledge, this is the first report of placentitis associated with propionibacteria in a cow.

Clinical and Laboratory Standards Institute interpretive breakpoints for in vitro susceptibility tests that predict mecA-mediated oxacillin resistance in Staphylococcus pseudintermedius isolates from animals have been changed twice in the past decade. Moreover, there are no counterpart recommendations for human isolates of S. pseudintermedius. Individual medical and veterinary laboratories variably use interpretive breakpoints identical to those recommended for use with Staphylococcus aureus or identical to those recommended for use with coagulase-negative staphylococci. The purpose of the current study was to examine correlations between oxacillin disk diffusion, oxacillin gradient diffusion, oxacillin microbroth dilution, and cefoxitin disk diffusion tests used to predict mecA-mediated resistance in S. pseudintermedius and to retrospectively estimate, from disk diffusion zone diameter measurements, the prevalence and rate of increase of oxacillin resistance among canine S. pseudintermedius isolates submitted to a veterinary teaching hospital laboratory. Oxacillin disk diffusion zone diameters of or=0.5 microg/ml were highly correlated with detection of mecA in canine S. pseudintermedius isolates by polymerase chain reaction. MecA-mediated resistance among S. pseudintermedius isolates from dogs increased from less than 5% in 2001 to near 30% in 2007. More than 90% of the methicillin-resistant S. pseudintermedius isolates in 2006 and 2007 were also resistant to representatives of >or=4 additional antimicrobial drug classes. Cefoxitin disk diffusion with the resistance breakpoint set at

Based on the scarcity of reports in veterinary medicine literature, it appears that Arcanobacterium spp. are rarely isolated from horses. Recently, a single isolate from vaginal discharge in a mare was characterized as a new species, Arcanobacterium hippocoleae. The present report describes a case of necrosuppurative placentitis and stillbirth in an American Quarterhorse mare. Numerous colonies of irregular, Gram-positive, rod-shaped bacteria were observed by histological examination within fibrin lattice associated with placental lesions. Arcanobacterium hippocoleae was isolated in large numbers from the placenta, lung, and stomach contents. To the authors' knowledge, this is the first report of placentitis associated with Arcanobacterium spp. in a horse and the third reported isolation of A. hippocoleae from a horse.

Reptiles are well-known sources of human Salmonella infections; however, little is known about the ability of Salmonella to cause disease in reptiles. Thirty-seven isolates of Salmonella enterica subspecies arizonae (S. arizonae) were obtained from retrospective and prospective studies of a closed colony of ridgenose rattlesnakes (Crotalus willardi) with osteomyelitis. All isolates (N = 7) from bone lesions were of a single serotype, 56:z4,z23, and this serotype was found on only 1 occasion among 8 other serotypes isolated from 21 cloacal and intestinal samples. The remainder (N = 7) of serotype 56:z4,z23 isolates were from other extraintestinal sites, including liver, ovary, blood, and testis. S. arizonae isolates were susceptible to most antimicrobials, and plasmid profiles did not correlate with serotype or antimicrobial resistance. Isolates of the 56:z4,z23 serotype (N = 14) formed a tight cluster with 95% similarity by XbaI macrorestriction analysis. Individual isolates of serotypes, 56:z4,z23, 38:(k)-z35, and 48:i-z invaded HeLa cells but an isolate of serotype 50:r-z did not. The same individual isolates of serotype 56:z4,z23 and 48:i-z also invaded viper heart cells. The Salmonella InvA gene was detected by polymerase chain reaction (PCR) in all S. arizonae serotypes tested, including 5 serotype 56:z4,z23 isolates and individual isolates of serotypes 48:i-z and 50:r-z. A source or possible explanation for increased virulence of S. arizonae serotype 56:z4,z23 in this unique host has not been found.

In this study, 2 reverse transcriptase-polymerase chain reaction (RT-PCR) assays were developed and compared for simultaneous detection of bovine and ovine respiratory syncytial viruses (RSVs). One assay was based on a set of primers, which amplified a 426-bp fragment of either bovine or ovine RSV F gene (RT-PCR F). The F products could be distinguished by EcoRI or BstYI restriction endonuclease cleavage. In the other assay, a set of primers amplified a 542-bp fragment of either ovine or bovine RSV G gene (RT-PCR G). EcoO1091 and RsaI restriction enzymes were used to differentiate between the ovine and bovine PCR-G products. Sequencing of the PCR products confirmed the fidelity of both assays. The 2 assays were evaluated using 18 bovine RSV isolates, 1 ovine RSV, 1 bighorn sheep RSV isolate, 1 caprine RSV isolate, 2 human RSV isolates, and several other viruses associated with bovine respiratory tract disease. RT-PCR G may be more sensitive in detecting viral RNA. Because the target sequence of the F gene is more conserved than that of the G gene, RT-PCR F followed by the appropriate restriction enzyme cleavage may be superior to RT-PCR G to discriminate between the 2 ruminant RSV subgroups. This assay should prove useful for determining the relative contribution of ovine and bovine RSV to the pathogenesis of bovine respiratory tract disease.

Ruminant respiratory syncytial viruses (RSVs) are classified into 2 subgroups, ovine RSV and bovine RSV. Although ovine RSV infects cattle, its contribution to bovine respiratory tract disease has not been established, which is an important issue for vaccine development in cattle. Diagnosis by virus isolation or serology has low or variable sensitivity and/or specificity and polymerase chain reaction (PCR) has been recommended as a rapid and sensitive technique for RSV detection. A simple procedure has been developed to detect and identify bovine and ovine RSVs. First, the nucleotide sequence of the ovine RSV fusion (F) gene was determined and compared with representative strains of bovine RSV and human RSV subgroups A and B. The ovine RSV F gene has 85 and 72-73% nucleotide identity with those of bovine RSV and human RSV, respectively. The predicted amino acid sequence of the ovine RSV F gene has 94 and 83-84% amino acid identity with those of bovine RSV and human RSV, respectively. Then PCR primers targeting a specific F gene fragment of bovine and ovine RSV were designed. The primers represented bases 85-103 and the complementary sequence to bases 510-493 of the ovine RSV F gene. A similar PCR product (426 bp) was obtained on agarose gel electrophoresis from bovine RSV and from ovine RSV. The products, however, were unique to the parent virus and could be distinguished by EcoRI or MspI restriction endonuclease cleavage. EcoRI cleaved the ovine product into 2 bands (285 and 141 bp) but failed to affect the bovine RSV PCR product. However, MspI cleaved the bovine product into 2 bands (229 and 197 bp) but had no effect on the ovine product. Also, this assay did not amplify any PCR product with human RSV. The reverse transcription-polymerase chain reaction (RT-PCR) followed by restriction enzyme digestion is a useful and practical approach for detection and differentiation of ruminant respiratory syncytial viruses.

Subgroup-specific peptide-based enzyme-linked immunosorbent assays from the G-protein of the ovine and bovine respiratory syncytial virus (RSV), respectively, were used to determine the prevalence of the ovine and bovine subgroup strains of RSV infections in cattle. A total of 1,102 bovine serum samples were obtained from 6 diagnostic laboratories located in the northwestern and the southeastern USA and were tested for antibody to either the bovine or ovine subgroups of RSV. Antibody to viruses from each subgroup was present in samples from each region and all states tested. The Southeast had a higher prevalence of the bovine subgroup strains (69.5%). Then did the Northwest (40.9%). The prevalence of the ovine strain was similar for the two regions (16.7% in the southeast, 14.9% in the northwest). The overall prevalence was 56.6% for the bovine strain and 15.9% for the ovine strain. These results suggest members of the ovine subgroup of RSV circulate in the cattle population but with less frequency than those viruses of the bovine subgroup.

Subgroup-specific peptide-based enzyme immunoassays from each respective G-glycoprotein of the ovine and the bovine respiratory syncytial virus (RSV) were developed to detect RSV-specific IgG responses in cattle. Antigenic peptides from the respective G-glycoprotein were identified from the extracellular central hydrophobic region (amino acids 158-189) located between 2 mucin-rich regions. These antigenic peptides identified by epitope mapping from each G-glycoprotein were synthesized and used to develop the subgroup-specific enzyme immunoassays. The negative cutoff for each enzyme immunoassay was established as the mean optical density of indirect immunofluorescent antibody-negative bovine sera plus 3 SDs. The sensitivity (82.9%) and specificity (100%) of the bovine enzyme immunoassay and the specificity (95.8%) of the ovine enzyme immunoassay were determined by comparison with indirect immunofluorescence (used as the "gold standard"). The negative and positive predictive values were calculated for each assay. The presence of serum antibody to ovine RSV in cattle implies that this virus infects cattle and may contribute to the pathogenesis of bovine respiratory disease.

An enzyme-linked immunosorbent assay (ELISA) based on synthetic peptides identical to lentivirus envelope protein amino acid sequences was used to study serologic reactivity of lentivirus-infected domestic cats and nondomestic felids. One feline immunodeficiency virus (FIV) peptide, P237, was consistently recognized by antibodies from FIV-infected cats, but 2 other FIV peptide antigens were not. The molecular basis for this serologic reactivity was examined. Lentivirus-infected nondomestic Felis species reacted intensely with a puma lentivirus (PLV) peptide corresponding to the conserved FIV peptide. However, lentivirus-infected Panthera species, from which a different lentivirus has been isolated, did not react with the PLV. FIV-infected domestic felids also did not have significant reactivity with the PLV peptide. The peptide ELISA is comparable in sensitivity and specificity to western blot analysis and a commercial enzyme immunoassay. Unlike the other assays, however, the peptide ELISA is inexpensive, requires a small amount of serum, enables the study of specific isotype reactivity, and discriminates between antibodies to FIV and those to PLV. Antibody tests based upon the FIV and the PLV peptides should be useful for detecting the possible introduction of FIV into exotic felids or of lentiviruses from nondomestic felids into the domestic cat population.