Evaluation of ketoconazole and itraconazole for treatment of disseminated cryptococcosis in cats

During the first part of a study, cats were inoculated with Cryptococcus neoformans via the following routes: intradermal, intranasal, IV, and intracisternal. Only use of the IV route of inoculation consistently induced disseminated cryptococcosis. In the second part of the study, disseminated cryptococcosis was experimentally induced in cats via IV inoculation of C neoformans. One month after inoculation, 3 cats were treated with ketoconazole (10 mg/kg of body weight/d) and 3 cats were treated with itraconazole (10 mg/kg/d) for 3 months. One of the ketoconazole-treated and 2 of the itraconazole-treated cats also had cryptococcosis of the CNS when treatment was begun. During treatment, serum cryptococcal antigen titer progressively decreased in all cats. Abnormalities in CBC values or the serum biochemical profile were not found in any cat during treatment. However, all ketoconazole-treated cats became anorectic and lost weight. Side effects were not seen in itraconazole-treated cats. During the 3-month posttreatment observation period, all cats remained healthy. At necropsy, histologic evidence of cryptococcosis was not found in the 3 ketoconazole-treated cats or in 2 of the itraconazole-treated cats. In the third itraconazole-treated cat, cryptococcal organisms were found in the kidneys.     

Peripheral vestibular disease associated with cryptococcosis in three cats.

Peripheral vestibular disease referable to otitis media/interna was the main reason for presentation in three cats with cryptococcosis. In two cats, Cryptococcus neoformans var neoformans was isolated from the tympanic bulla. In the remaining cat, otitis media/interna was considered to be secondary to occlusion of the auditory tube by a nasopharyngeal granuloma associated with a C neoformans var gattii infection. This report emphasises the importance of maintaining an index of suspicion for a fungal aetiology in cats with signs of otitis media/interna, particularly in countries with a high prevalence of cryptococcosis. The presence of C neoformans may be overlooked with potentially fatal consequences where only standard methods for bacterial isolation are used to examine samples obtained from the middle ear.     

Cryptococcosis in seven horses

The clinical, radiographic and post-mortem findings in 6 horses with cryptococcal pneumonia and one horse with an abdominal cryptococcal granuloma are described. In pulmonary cryptococcosis, the lesions were either diffuse and multiple, with bilateral lung involvement, or localised mainly to the dorsocaudal region of one lung. The cases of diffuse multiple cryptococcosis were thought to be associated with haematogenous spread of the fungus after gastrointestinal infection and dissemination from regional lymph nodes. The localised form of the disease was thought to have been associated with inhalation of cryptococci. In all cases of pulmonary cryptococcosis, encapsulated yeast-like organisms were demonstrated in Wright's-stained sediment of tracheal washes. In the horse with the abdominal granuloma, cryptococci were present in a fine needle aspirate sample. Isolates of Cryptococcus neoformans var gattii were recovered from 2 of the 5 horses in which cultures were attempted. In addition to a history of previous illness that may have predisposed to infection, most horses in this report had been in areas in which Eucalyptus camaldulensis, or the closely related E rudis, were growing. In humans, an epidemiological relationship between E camaldulensis and infection with C neoformans var gattii has been suggested. Cases of equine cryptococcosis carry a poor prognosis and treatment was not attempted in any of these cases.     

Cryptococcosis in captive cheetah (Acinonyx jubatus): two cases

Cryptococcus neoformans is a yeast-like organism associated with pulmonary, meningoencephalitic, or systemic disease. This case report documents 2 cases of cryptococcosis with central nervous system involvement in captive cheetah (Acinonyx jubatus). In both cases the predominant post mortal lesions were pulmonary cryptococcomas and extensive meningoencephalomyelitis. Both cheetahs tested negative for feline immunodeficiency virus and feline leukaemia virus. The organism isolated in Case 2 was classified as Cryptococcus neoformans var. gattii, which is mainly associated with disease in immunocompetent hosts.     

Itraconazole for the Treatment of Cryptococcosis in Cats

Itraconazole was used in 35 cats with cryptococcosis. Treatment response was determined by comparing clinical signs before, during, and after treatment. It could not be evaluated in 7 cats because they died during treatment from causes unrelated to cryptococcosis. Of the remaining 28 cats, treatment response was classified as success in 16 cats (57%), as improvement in 8 cats (29%), and as a failure in 4 (14%). The failures were due to death or euthanasia from drug toxicity (1 cat), progressive fungal disease (2 cats), and relapse 1 year after treatment (1 cat). The cats that improved did not undergo a 1 -year posttreatment evaluation because they were lost to follow-up (3 cats), died or were euthanatized for other reasons (4 cats), or had a noncompliant owner (1 cat). For the 16 cats in which treatment was successful, the median itraconazole dose was 13.8 mg/kg body weight daily (range, 10.9 to 26.7 mg/kg/d), and the median duration of treatment was 8.5 months (range, 4 to 16 months). Five of these cats had previously been treated unsuccessfully with ketoconazole.     

Babesiosis in a litter of pups

Babesia canis infection was diagnosed in a litter of seven 3-week-old Mastiff pups kept in a north Florida kennel. The pups were evaluated because of poor weight gain; the smallest pup also was markedly lethargic. Six of the pups were anemic and thrombocytopenic. A positive linear correlation between PCV and absolute reticulocyte count suggested that the variation in PCV may have been related more to the ability of a pup to increase erythrocyte production than to a difference in magnitude of erythrocyte destruction. All pups recovered from clinical signs and hematologic abnormalities attributable to babesiosis within 2.5 weeks after treatment with diminazene aceturate. Transient neurologic signs observed in 1 pup 3 days after treatment were believed to represent an adverse drug reaction. The dam of the litter had a serum titer of 1:640 for B canis, but appeared healthy, as did approximately 30 other adult dog in the kennel. The strain of B canis infecting dogs in the kennel caused severe illness and death in some pups, but clinically inapparent disease in adult dogs.     

Cryptococcosis in ferrets: a diverse spectrum of clinical disease

Cryptococcosis was diagnosed in seven ferrets (five from Australia; two from western Canada) displaying a wide range of clinical signs. Two of the ferrets lived together. One (5-years-old) had cryptococcal rhinitis and presented when the infection spread to the nasal bridge. Its sibling developed cryptococcal abscessation of the right retropharyngeal lymph node 12 months later, soon after developing a severe skin condition. DNA fingerprinting and microsatellite analysis demonstrated that the two strains isolated from these siblings were indistinguishable. Two ferrets (2- to 3-years-old) developed generalised cryptococcosis: one had primary lower respiratory tract disease with pneumonia, pleurisy and mediastinal lymph node involvement, while in the other a segment of intestine was the primary focus of infection with subsequent spread to mesenteric lymph nodes, liver and lung. The remaining three ferrets (1.75 to 4-years-old) had localised disease of a distal limb, in one case with spread to the regional lymph node. Cryptococcus bacillisporus (formerly C. neoformans var gattii) accounted for three of the four infections in Australian ferrets where the biotype could be determined. The Australian ferret with intestinal involvement and the two ferrets from Vancouver had C. neoformans var grubii infections.     

Diagnosis of systemic cryptococcosis by fecal cytology in a dog

A 3-year-old Boxer was presented with progressive diarrhea, vomiting, and lethargy of 5-months duration. The dog had watery black feces, a mature neutrophilia, and microcytic anemia. Cytologic evaluation of a direct fecal smear stained with Wright's-Giemsa revealed numerous encapsulated, narrow-based, budding organisms consistent with Cryptococcus sp. Pyogranulomatous inflammation and Cryptococcus organisms also were observed in ultrasound-guided fine-needle aspirates of the small intestine and mesenteric lymph nodes, and in histologic sections of colonic biopsies obtained by endoscopy. Multifocal chorioretinitis by fundic examination was consistent with systemic mycosis, and the reciprocal antigen titer (1600) on a cryptococcal antigen latex agglutination test for Cryptococcus neoformans was markedly increased. Using immunohistochemistry, the organism was identified further as C neoformans var. grubii (C neoformans var. neoformans serotype A). After 3 weeks of antifungal treatment, ultrasound examination revealed urinary bladder wall thickening, and Cryptococcus organisms were found in a urine sediment preparation. After 4 months of treatment, the dog was clinically normal and had no abnormal findings on CBC, serum biochemistry, urinalysis, or fecal cytology; however, the antigen titer remained unchanged, mesenteric lymphadenomegaly and jejunal wall thickening were still evident, and cytologic evaluation of fine-needles aspirates of the jejunal wall revealed budding Cryptococcus organisms. Intestinal involvement in dogs with cryptococcosis is rare, and diagnosis by fecal cytology has not been documented previously.     

Isolation of saprophytic Cryptococcus neoformans

Isolation of Cryptococcus neoformans was carried out on sunflower seed agar medium (SFA) and Sabouraud dextrose agar (SDA). Out of 346 environmental substrates (133 fruits, 107 avian extreta, 91 vegetables and 15 wooden scrapings) tested, 3 specimens were positive for C. neoformans. The positive isolations came from the fruits of 2 banana (Musa sapientum) and a potato tuber (Solnum tuberosum). The pathogen could not be demonstrated in 107 samples of avian droppings and 15 of wooden materials. All the 3 isolates of the yeast were obtained on SFA, while they were not cultured on the plates of SDA with chloramphenicol which were badly contaminated with rapidly growing molds, yeasts and bacteria. To the present author's knowledge, this appears to be the first reports of the isolation of this pathogenic basidiomycetous yeast from contaminated fruits of banana. We suggest more comprehensive ecological surveys to search for environmental niche of C. neoformans var. neoformans and C. neoformans var. gattii as the latter variety is also implicated in the etiology of cryptococcosis.     

Serotypes and mating types of clinical isolates from feline cryptococcosis in Japan

Most isolates of Cryptococcus neoformans (teleomorph: Filobasidiella neoformans) from human patients and from environmental materials in Japan have been identified as serotype A mating type a by the seroagglutination test and mating experiments. A PCR method using the mating type alpha allele-specific primer of the STE12 gene and the serotype- and mating type-specific primers of the STE20 gene for identification of C. neoformans has been developed. Using the PCR method, conserved strains and clinical isolates from feline cryptococcosis were examined for serotype and the mating type. The results showed that all clinical isolates examined were identified as serotype A, MATalpha, indicating that feline cryptococcsis cases in Japan are caused by C. neoformans serotype A, MATalpha, as is the case in humans.     

Triploidy (117,XXX) in a stillborn canine pup conceived with frozen semen

Analysis of skin fibroblast cultures from a deformed stillborn female Alaskan Malamute pup revealed a 3n = 117,XXX (normal, 2n = 78,XX) chromosome count. The triploid pup was delivered by cesarean section 5 days after estimated date of parturition, because the bitch failed to start labor. The bitch had been inseminated with thawed frozen semen deposited into the lumen of the uterus approximately 4 days after ovulation. Gross anatomic abnormalities of the pup included omphalocele, diaphragmatic hernia, persistent right aortic arch, atresia ani, and no right forelimb distal to the scapula. The pup also had arthrogryposis of the left carpus, kyphosis of the thoracic vertebrae, widely separated cranial sutures, open fontanelles, hydrocephalus, and cleft palate. Suspected cause of the triploidy was dispermy of an aged oocyte after intrauterine deposition of the thawed frozen semen. Numeric chromosome abnormalities may be causes of fetal deformity or death that can be detected by fetal karyotype.     

PCR detection of the Cryptococcus neoformans CAPS9 gene from a biopsy specimen from a case of feline cryptococcosis.

A polymerase chain reaction assay was developed to detect Cryptococcus neoformans in biopsy samples. The assay detects the CAP59 gene of Cryptococcus neoformans and was used to substantiate cutaneous cryptococcosis in a 5-year-old cat submitted to the Veterinary Medicine Center at the University of Tokyo.     

Canine parvoviral myocarditis

A 5-week-old crossbred pigdog pup was presented in respiratory distress. It appeared to dry retch, and then died suddenly before a clinical examination was performed; a littermate had died suddenly the week before. Histopathological examination revealed an ongoing lymphocytic myocarditis, characterised by myofibril degeneration and lymphocytic inflammatory infiltrates. In scattered degenerate myofibres, there were large amphophilic intranuclear inclusion bodies distending their nuclei. There was also marked pulmonary oedema. Canine parvoviral myocarditis, due to canine parvovirus type 2, was diagnosed. Further questioning regarding the history of this pup revealed that it had been born to an unvac-cinated dam that had been moved late in her pregnancy from an isolated farm on to another property. It seems likely that the dam was naïve and only became exposed to the virus immediately prior to whelping. The lack of wildlife vectors and a well-controlled population of dogs in rural New Zealand means that there may be isolated areas of the country where dogs can reach maturity without becoming exposed to parvovirus, creating the potential for development of this now uncommon syndrome. When parvovirus first arose in New Zealand, between 1979–;1981, parvoviral myocarditis was relatively common, occurring in neonates when the virus was introduced to breeding kennels that previously had been unexposed.     

Clinicopathologic features of an unusual outbreak of cryptococcosis in dogs, cats, ferrets, and a bird: 38 cases (January to July 2003)

OBJECTIVE: To determine clinical and pathologic findings associated with an outbreak of cryptococcosis in an unusual geographic location (British Columbia, Canada). DESIGN: Retrospective study. ANIMALS: 1 pink-fronted cockatoo, 2 ferrets, 20 cats, and 15 dogs. PROCEDURE: A presumptive diagnosis of cryptococcosis was made on the basis of serologic, histopathologic, or cytologic findings, and a definitive diagnosis was made on the basis of culture or immunohistochemical staining. RESULTS: No breed or sex predilections were detected in affected dogs or cats. Eleven cats had neurologic signs, 7 had skin lesions, and 5 had respiratory tract signs. None of 17 cats tested serologically for FeLV yielded positive results; 1 of 17 cats yielded positive results for FIV (western blot). Nine of 15 dogs had neurologic signs, 2 had periorbital swellings, and only 3 had respiratory tract signs initially. Microbiologic culture in 15 cases yielded 2 isolates of Cryptococcus neoformans var grubii (serotype A) and 13 isolates of C. neoformans var gattii (serotype B); all organisms were susceptible to amphotericin B and ketoconazole. Serologic testing had sensitivity of 92% and specificity of 98%. CONCLUSIONS AND CLINICAL RELEVANCE: Serologic titers were beneficial in identifying infection in animals with nonspecific signs, but routine serum biochemical or hematologic parameters were of little value in diagnosis. Most animals had nonspecific CNS signs and represented a diagnostic challenge. Animals that travel to or live in this region and have nonspecific malaise or unusual neurologic signs should be evaluated for cryptococcosis.     

Testosterone concentrations,testes weight and morphology of mule drakes (Muscovy drake X Khaki Campbell)

1. Mule ducks were produced by naturally mating Muscovy drakes and Khaki Campbell ducks. 2. Semen was collected from 6-month-old mule drakes via an artificial vagina. The fluid was clear without any spermatozoa or spermatids. 3. Testes from 27-week-old mule drakes were smaller in size than those of Khaki Campbell drakes but heavier than Muscovy males of the same age. Histological sections of these testes revealed that spermatogenesis was not complete. 4. Testosterone concentration in the mule drakes was higher than in Muscovy males but similar to Khaki Campbell drakes. 5. Mule drakes have strong sexual drives as a result of high concentrations of testosterone, but, because spermatogenesis is incomplete, their semen had no sperm.     

Prediction of Parturition in Bitches Utilizing Continuous Vaginal Temperature Measurement

The objective of this study was to determine sensitivity and specificity of a body temperature decline in bitches to predict parturition. Temperature loggers were placed into the vaginal cavity of 16 pregnant bitches on day 56–61 after estimated ovulation or first mating. This measurement technique has been validated previously and enabled continuous sampling of body temperature. The temperature loggers were expelled from the vagina before delivery of the first pup. The computed values for specificity (77–92%) were higher than sensitivity (53–69%), indicating a more precise prognosis of parturition not occurring. In conclusion, our findings may assist interpreting vaginal temperature measurements in order to predict parturition in bitches.     

Disseminate Cryptococcosis in a guenon (Cercopithecus ascanius)

A 23-year-old captive-bred red-tailed guenon (Cercopithecus ascanius) with a brief history of inappetence, lethargy, and seizures was submitted for necropsy. On postmortem examination, multiple cryptococcomas were identified in brain and heart. Cryptococcus neoformans organisms were also identified microscopically in kidney, eye, and pancreas. Fungal yeast formed rare pseudohyphae. The histologic diagnosis of cryptococcosis was confirmed by a positive test for C. neoformans antigen in a serum sample. Immunohistochemical staining confirmed that macrophages were the principal inflammatory cell in brain lesions and often contained phagocytosed yeast. As disseminate cryptococcosis is often associated with immune suppression, serology and immunohistochemical staining for simian immunodeficiency virus were performed but showed no evidence of SIV infection.     

Detection of CAP59 gene in 2 feline cases of systemic cryptococcosis

Two feline cases were diagnosed as systemic cryptococcosis due to Cryptococcus neoformans (teleomorph: Filobasidiella neoformans) by PCR assay with CAP59 gene primers using urine, serum and biopsy samples. The results of molecular analysis were consistent with the mycological findings.     

Infections with Encephalitozoon cuniculi and Leptospira interrogans, serovars grippotyphosa and ballum, in a kennel of foxhounds

Leptospira interrogans serovars grippotyphosa and ballum were isolated from kidney and urine of an American Foxhound pup. The pup was from a litter of 12, all of which were unthrifty. Titers for serovar grippotyphosa in pups from the litter ranged from 200 to 6,400 and 23 of 36 adult dogs in the kennel had titers to that serovar. None of the sera was tested for antibodies to serovar ballum. Leptospires were not isolated from or observed in 2 littermates and 1 penmate, but gram-positive organisms morphologically compatible with Encephalitozoon cuniculi were detected in their brains and kidneys.     

Cryptococcosis of the central nervous system in a dog

A 3-year-old female German Shepherd Dog was evaluated for progressive mental obtundation and vestibular signs. Central nervous system cryptococcosis was diagnosed on the basis of growth of Cryptococcus neoformans in fungal culture of CSF, as well as detection of the organism in CSF via microscopy. Cryptococcal capsular latex antigen agglutination titer was 1:262,144 in CSF and 1:1,048,576 in serum samples. Imaging with magnetic resonance augmented diagnosis. The dog improved after long-term treatment with fluconazole. Fluconazole is useful in the treatment of CNS cryptococcosis, because it attains high concentration in the CNS. Long-term therapy is often required for resolution of clinical signs, and affected animals may require long-term follow-up with periodic evaluation of CSF via fungal culture and latex agglutination tests. Monitoring serum latex agglutination test results may provide a safe, less invasive means of monitoring response to treatment.