Colorectal basidiobolomycosis in a dog.

A 7-year-old castrated male French Bulldog was examined for chronic large intestinal enteropathy. A colonic mass and thickened rectal mucosa were identified, and histopathologic examination of endoscopic biopsy specimens disclosed eosinophilic proctitis with large (5-20 µm), irregularly shaped, pauciseptate hyphae that were Gomori methenamine silver and periodic acid-Schiff positive. Amplification and sequencing of ribosomal DNA extracted from paraffin-embedded tissues yielded a sequence with 97% identity to GenBank sequences for Basidiobolus ranarum. After itraconazole, terbinafine, and prednisone administration, clinical signs resolved rapidly, and sonographic lesions were largely absent after 6 weeks. Treatment was discontinued by the owner 15 weeks after diagnosis. Three weeks later, the dog collapsed acutely and was euthanized. Necropsy identified metastatic islet cell carcinoma and grossly unremarkable colorectal tissues. However, histopathology of the rectum disclosed multifocal submucosal granulomas with intralesional hyphae morphologically similar to those previously observed. This report is the first to describe medical treatment of gastrointestinal basidiobolomycosis in a dog.

Trapped Neutrophil Syndrome in a Border Collie.

A 10 wk old female border collie was presented for hemorrhagic diarrhea and pelvic limb lameness. Examination revealed pain and effusion in multiple appendicular joints and pyrexia. Clinicopathologic testing revealed moderate neutropenia as well as nondegenerate neutrophilic inflammation in multiple joints. Radiographs showed capsular joint swelling and heterogeneous metaphyseal lucencies in the distal radius, ulna, femur, and tibia. Genetic testing confirmed a mutation in the vacuolar protein sorting-associated protein 13B gene and a diagnosis of trapped neutrophil syndrome (TNS). Within 24 hr of initiating prednisone therapy (1 mg/kg, per os, q 12 hr), the dog was afebrile and nonpainful with normal ambulation. Lameness recurred twice over the next 5 mo. At 9 mo of age, diagnostics showed severe erosive polyarthritis of both stifles with an inflammatory leukogram and arthrocentesis findings consistent with septic arthritis, and the dog died despite antibiotic therapy. This is the first case of TNS described in the North American literature, and it is unique in that we had the opportunity to document progression of radiographic abnormalities over more than 6 mo. TNS should be considered in young border collies with signs suggestive of immune-mediated polyarthritis, septic arthritis, or hypertrophic osteodystrophy, combined with neutropenia or gastrointestinal signs.

Successful management of 3 dogs with colonic pythiosis using itraconzaole, terbinafine, and prednisone.

Gastrointestinal (GI) pythiosis is a severe and often fatal disease in dogs that traditionally has been poorly responsive to medical treatment. Although aggressive surgical resection with wide margins is the most consistently effective treatment, lesion location and extent often preclude complete resection. Recently, it has been suggested that the addition of anti-inflammatory doses of corticosteroids may improve outcome in dogs with nonresectable GI pythiosis. This report describes 3 dogs with colonic pythiosis in which complete resolution of clinical signs, regression of colonic masses, and progressive decreases in serological titers were observed after treatment with itraconazole, terbinafine, and corticosteroids. This treatment protocol represents a promising treatment for dogs with GI pythiosis in which surgical intervention is not feasible.

Quantitation of anti-Pythium insidiosum antibodies before and after administration of an immunotherapeutic product to healthy dogs.

OBJECTIVE To evaluate the effect of an immunotherapeutic product on concentrations of anti-Pythium insidiosum antibodies in dogs. ANIMALS 7 healthy hound-crossbreds. PROCEDURES Antibody concentrations were evaluated before (day 0) and after administration of the immunotherapeutic product. The immunotherapeutic product was administered on days 0, 7, and 21. Serum was obtained on days 0, 7, 14, 21, 28, 35, 42, 49, and 56. Anti-P insidiosum antibody concentrations were measured and reported as the percentage positivity relative to results for a strongly positive control serum. RESULTS Mean ± SD percentage positivity before administration of the immunotherapeutic product was 7.45 ± 3.02%. There was no significant change in anti-P insidiosum antibody concentrations after administration of the product, with percentage positivity values in all dogs remaining within the range expected for healthy dogs (3% to 15%). CONCLUSIONS AND CLINICAL RELEVANCE Administration of the immunotherapeutic product to healthy dogs in accordance with the manufacturer's suggested protocol did not induce a significant change in anti-P insidiosum antibody concentrations. These results suggested that administration of the immunotherapeutic product may not interfere with postadministration serologic monitoring. However, further investigations will be required to determine whether there is a similar effect in naturally infected dogs.

Molecular phylogeny and taxonomy of Lagenidium-like oomycetes pathogenic to mammals.

Over the past twenty years, infections caused by previously unrecognised oomycete pathogens with morphological and molecular similarities to known Lagenidium species have been observed with increasing frequency, primarily in dogs but also in cats and humans. Three of these pathogens were formally described as Lagenidium giganteum forma caninum, Lagenidium deciduum, and Paralagenidium karlingii in advance of published phylogenetic verification. Due to the complex nature of Lagenidium taxonomy alongside recent reports of mammalian pathogenic species, these taxa needed to be verified with due consideration of the available data for Lagenidium and its allied genera. This study does so through morphologic characterisation of the mammalian pathogenic species, and phylogenetic analyses. The six-gene phylogeny generally supports the most recent comprehensive classification of Lagenidium with a well-supported Lagenidium clade that includes the mammalian pathogens L. giganteum f. caninum and L. deciduum, and well-supported clades for which the names Myzocytiopsis and Salilagenidium can be applied. The genus Paralagenidium is phylogenetically unrelated to any of the main clades within the class Peronosporomycetes. Close relationships between pathogens of mammals and those of insects or nematodes were revealed. Further characterisation of Lagenidium-like taxa is needed to establish the risk of mammalian infection by pathogens of insects and nematodes.

Protein A/G-based immunochromatographic test for serodiagnosis of pythiosis in human and animal subjects from Asia and Americas.

Pythiosis is a life-threatening infectious disease of both humans and animals living in Asia, Americas, Africa, and parts of Australia and New Zealand. The etiologic pathogen is the fungus-like organism Pythium insidiosum The disease has high mortality and morbidity rates. Use of antifungal drugs are ineffective against P. insidiosum, leaving radical surgery the main treatment option. Prompt treatment leads to better prognosis of affected individuals, and could be achieved by early and accurate diagnosis. Since pythiosis has been increasingly reported worldwide, there is a need for a rapid, user-friendly, and efficient test that facilitates the diagnosis of the disease. This study aims to develop an immunochromatographic test (ICT), using the bacterial protein A/G, to detect anti-P. insidiosum IgGs in humans and animals, and compare its diagnostic performance with the established ELISA. Eighty-five serum samples from 28 patients, 24 dogs, 12 horses, 12 rabbits, and 9 cattle with pythiosis, and 143 serum samples from 80 human and 63 animal subjects in a healthy condition, with thalassemia, or with other fungal infections, were recruited for assay evaluation. Detection specificities of ELISA and ICT were 100.0%. While the detection sensitivity of ELISA was 98.8%, that of ICT was 90.6%. Most pythiosis sera, that were falsely read negative by ICT, were weakly positive by ELISA. In conclusion, a protein A/G-based ICT is a rapid, user-friendly, and efficient assay for serodiagnosis of pythiosis in humans and animals. Compared to ELISA, ICT has an equivalent detection specificity and a slightly lower detection sensitivity.

Fungal myocarditis and pericardial effusion secondary to Inonotus tropicalis (phylum Basidiomycota) in a dog.

Fungal disease is a rare cause of pericardial effusion in dogs. This report describes the first case of fungal pericardial effusion and myocarditis secondary to the fungal organism Inonotus tropicalis. A 9-year-old female spayed French bulldog with a multi-year history of treatment with glucocorticoids for management of atopy was presented for exercise intolerance, ascites and weight loss. Physical examination and thoracic imaging revealed enlarged peripheral and cranial mediastinal lymph nodes, left ventricular thickening and cardiac tamponade secondary to pericardial effusion. Fine needle aspiration of the cranial mediastinal lymph node showed pyogranulomatous inflammation with short, thin and poorly septated hyphae. Culture of the aspirate yielded a fungal isolate identified as Inonotus tropicalis based on morphologic features and rRNA gene sequencing. Postmortem examination showed myocardial thickening with multifocal to coalescing, firm, white, ill-defined nodules. Histology confirmed the presence of disseminated fungal infection with extensive myocardial involvement. Inonotus tropicalis is an opportunistic poroid wood-decaying basidiomycete. Infection in this dog was likely the result of chronic immunosuppressive therapy.

Dual infection with Pythium insidiosum and Blastomyces dermatitidis in a dog.

A 4-year-old male neutered Labrador Retriever with severe gastrointestinal signs, but no respiratory signs, was diagnosed with multifocal pyogranulomatous gastritis, enteritis, and lymphadenitis with intralesional hyphae and multifocal pyogranulomatous pneumonia with intralesional yeast. Based on cytologic evaluation, histologic examination with special stains, and immunohistochemical analysis of tissues collected antemortem or at necropsy, dual infections with Pythium insidiosum and Blastomyces dermatitidis were detected and are reported for the first time.

Fungal myelitis caused by Phialosimplex caninus in an immunosuppressed dog.

A bone marrow infection caused by Phialosimplex caninus was diagnosed in a seven-year-old female spayed Cocker Spaniel that was receiving prednisone for autoimmune hemolytic anemia. Histopathologic examination of a bone marrow core biopsy revealed clusters of oval to round yeast-like cells of varying shape and size and occasional irregular hyphae. Culture of a bone marrow aspirate sample yielded a mould initially suggestive of Paecilomyces inflatus or Sagenomella species but later determined to be P. caninus. The dog was treated with itraconazole and amphotericin B, and prednisone was continued at the lowest dose needed to control the hemolytic anemia. The patient died after 18 months of treatment. This is the first detailed clinical report of infection caused by P. caninus, a newly described fungus associated with disseminated disease in dogs.

Diagnosis and treatment of truncal cutaneous pythiosis in a dog.

CASE DESCRIPTION: A 4-year-old spayed female Boxer was evaluated for a cutaneous mass located on the dorsum. The mass had been present for 6 weeks and was increasing in size. CLINICAL FINDINGS: A mass of approximately 10 cm in diameter was detected on the dorsum cranial to the right ilial wing. Histologic examination of a tissue sample from the mass led to the diagnosis of cutaneous pythiosis. Computed tomography of the abdomen and the mass were performed and revealed a contrast-enhancing soft tissue mass of the dorsum and enlarged intra-abdominal lymph nodes. TREATMENT AND OUTCOME: The dog underwent surgical excision of the cutaneous mass, including 5-cm skin margins and deep margins of 2 fascial planes. The mass was completely excised on the basis of results of histologic examination of surgical margins. The dog received itraconazole and terbinafine by mouth for 3 months following surgery. Recheck examination at 20 months postoperatively showed no signs of recurrence of pythiosis at the surgical site. CLINICAL RELEVANCE: Aggressive surgical excision in combination with medical treatment resulted in a favorable long-term (> 1 year) outcome in this dog. Thorough workup including diagnostic imaging and lymph node evaluation is recommended. If surgery is to be performed, skin margins of 5 cm and deep margins of 2 fascial planes are recommended.

Assessment of exposure to Leptospira serovars in veterinary staff and dog owners in contact with infected dogs.

OBJECTIVE: To assess patterns of seroreactivity to Leptospira serovars in veterinary professional staff and dog owners exposed to dogs with acute leptospirosis and to contrast these patterns in people with those observed in dogs. DESIGN: Cross-sectional study. SAMPLE POPULATION: Human subjects consisted of 91 people (50 veterinarians, 19 technical staff, 9 administrative personnel, and 13 dog owners) exposed to dogs with leptospirosis. Canine subjects consisted of 52 dogs with naturally occurring leptospirosis admitted to the University of Bern Vetsuisse Faculty Small Animal Clinic in 2007 and 2008. PROCEDURES: People were tested for seroreactivity to regionally prevalent Leptospira serovars by use of a complement fixation test. A questionnaire designed to identify risk factors associated with seropositivity was used to collect demographic information from each study participant. Dogs were tested for seroreactivity to Leptospira serovars by use of a microscopic agglutination test. RESULTS: On the basis of microscopic agglutination test results, infected dogs were seropositive for antibodies against Leptospira serovars as follows (in descending order): Bratislava (43/52 [83%]), Australis (43/52 [83%]), Grippotyphosa (18/52 [35%]), Pomona (12/52 [23%]), Autumnalis (6/52 [12%]), Icterohemorrhagiae (4/52 [8%]), Tarassovi (2/52 [4%]), and Canicola (1/52 [2%]). All 91 people were seronegative for antibodies against Leptospira serovars. Therefore, statistical evaluation of risk factors and comparison of patterns of seroreactivity to Leptospira serovars between human and canine subjects were limited to theoretical risks. CONCLUSIONS AND CLINICAL RELEVANCE: Seroreactivity to Leptospira serovars among veterinary staff adhering to standard hygiene protocols and pet owners exposed to dogs with acute leptospirosis was uncommon.

Pharmacokinetics of voriconazole after oral administration of single and multiple doses in Hispaniolan Amazon parrots (Amazona ventralis).

OBJECTIVE: To determine the pharmacokinetics and safety of voriconazole administered orally in single and multiple doses in Hispaniolan Amazon parrots (Amazona ventralis). ANIMALS: 15 clinically normal adult Hispaniolan Amazon parrots. PROCEDURES: Single doses of voriconazole (12 or 24 mg/kg) were administered orally to 15 and 12 birds, respectively; plasma voriconazole concentrations were determined at intervals via high-pressure liquid chromatography. In a multiple-dose trial, voriconazole (18 mg/kg) or water was administered orally to 6 and 4 birds, respectively, every 8 hours for 11 days (beginning day 0); trough plasma voriconazole concentrations were evaluated on 3 days. Birds were monitored daily, and clinicopathologic variables were evaluated before and after the trial. RESULTS: Voriconazole elimination half-life was short (0.70 to 1.25 hours). In the single-dose experiments, higher drug doses yielded proportional increases in the maximum plasma voriconazole concentration (C(max)) and area under the curve (AUC). In the multiple-dose trial, C(max), AUC, and plasma concentrations at 2 and 4 hours were decreased on day 10, compared with day 0 values; however, there was relatively little change in terminal half-life. With the exception of 1 voriconazole-treated parrot that developed polyuria, adverse effects were not evident. CONCLUSIONS AND CLINICAL RELEVANCE: In Hispaniolan Amazon parrots, oral administration of voriconazole was associated with proportional kinetics following administration of single doses and a decrease in plasma concentration following administration of multiple doses. Oral administration of 18 mg of voriconazole/kg every 8 hours would require adjustment to maintain therapeutic concentrations during long-term treatment. Safety and efficacy of voriconazole treatment in this species require further investigation.

In vitro susceptibility of Pythium insidiosum and a Lagenidium sp to itraconazole, posaconazole, voriconazole, terbinafine, caspofungin, and mefenoxam.

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 microg/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 microg/mL. Terbinafine at 8 microg/mL significantly reduced growth of P insidiosum and at > or = 1 microg/mL significantly reduced growth of the Lagenidium sp. Caspofungin and mefenoxam (concentrations > or = 1 microg/mL and > or = 0.025 microg/mL, respectively) significantly reduced growth of both pathogens. Mefenoxam at 0.1 microg/mL caused > 50% growth inhibition in 11 of 12 isolates and at 1 microg/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.

Cutaneous pythiosis in a nontravelled California horse.

An 18-year-old Arabian mare was examined with a large mass on the left hind pastern and fetlock. The mare was located in the Central Valley of northern California, and had never been out of the state. Routine histopathological processing and examination of biopsy samples from the mass showed several hyphal organisms that were delineated with a silver stain. Using immunohistochemistry the organism was diagnosed as Pythium insidiosum. The owner declined debulking surgery, and despite treatment with an immunotherapeutic vaccine, the horse's condition deteriorated leading to euthanasia.

Hypercalcemia associated with gastric pythiosis in a dog.

A 20-month-old castrated male Labrador Retriever with a 3-month history of anorexia, weight loss, and vomiting was evaluated. Plasma biochemical abnormalities included marked hyperglobulinemia and hypercalcemia. Serum levels of parathyroid hormone, parathyroid hormone-related protein, 25-hydroxyvitamin D, and 1,25-dihydroxyvitamin D were either low or within reference intervals. Gastric wall thickening and abdominal lymphadenomegaly were observed with abdominal ultrasonography. Cytologic evaluation of a sample obtained via fine-needle aspiration of the gastric wall revealed pyogranulomatous inflammation and numerous poorly stained hyphae. Partial gastrectomy was performed, and a diagnosis of gastric pythiosis was made by immunohistochemical staining of infected gastric tissue, as well as by immunoblot serology. This case demonstrates that diagnostic samples for cytologic evaluation can be obtained by fine-needle aspiration of Pythium insidiosum-infected tissues and that a presumptive diagnosis can be made by examination of a Romanowsky-stained smear. Furthermore, pythiosis should be considered as a differential diagnosis for hypercalcemia, especially in young dogs with inflammatory lesions that have a granulomatous component. The mechanism for the hypercalcemia in this dog was not determined; however, calcium concentrations normalized after surgical resection of the gastric lesion.

Gastrointestinal pythiosis in two cats.

Two young adult male Domestic Shorthair cats living in the southeastern United States were evaluated for signs attributable to partial intestinal obstruction. Physical examination indicated a palpable abdominal mass in each animal. Exploratory laparotomy revealed a large extraluminal mass involving the ileum and mesentery with adjacent mesenteric lymphadenopathy in cat No. 1 and an abscessed mass in the distal duodenum in cat No. 2. Mass resection and intestinal anastomosis were performed in both cats. Histologic evaluation indicated that the intestinal lesions involved primarily the outer smooth muscle layer and serosa and consisted of eosinophilic granulomatous inflammation with multifocal areas of necrosis. In Gomori methenamine silver-stained sections, broad (2.5-7.5 microm), occasionally branching, infrequently septate hyphae were observed within areas of necrosis. A diagnosis of Pythium insidiosum infection was confirmed in both cats by immunoblot serology and by immunoperoxidase staining of tissue sections using a P. insidiosum-specific polyclonal antibody. Cat No. 1 was clinically normal for 4 months after surgery but then died unexpectedly from an unknown cause. Cat No. 2 has been clinically normal for at least 9 months after surgery and appears to be cured on the basis of follow-up enzyme-linked immunosorbent assay serology.