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CRYPTOCOCCOSIS IN STRANDED MARINE
MAMMALS IN BRITISH COLUMBIA
Martin Haulena, Justin Rosenberg, Muhammad
Morshed, Linda Hoang, Erin Zabek, Chelsea
Himsworth, Chelsea Anderson, Lindsaye Akhurst,
Marina Ivančić, Dean A. Malpas, John S. Pollock, and
Stephen Raverty
Vancouver Aquarium
British Columbia Provincial Animal Health Center
AquaVetRad
British Columbia Centre for Disease Control
Canada Diagnostic Centre
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Cryptococcus gattii
• Since 2000, there has been a recognized outbreak of cryptococcosis in human and companion animals on Vancouver Island, BC
– From early 1999 to mid 2000, microbiologist on Vancouver Island identified 19 cases of cryptococcosis in humans.
• Most individuals had no evidence of immunosuppression
– Central Laboratory for Veterinarians, Langley, BC reported 12 cases of cryptococcosis in companion animals on Vancouver Island for 2000
• Isolates typed as var gatti type B
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Case Summary Information• Porpoises presenting with
cryptococcosis
• Species– 10 Dall’s porpoises (Phocoenoides
dalli)
– 14 harbor porpoises (Phocoena phocoena)
– 1 Pacific white sided dolphin (Lagenorhynchus obliquidens)
• Gender– 10 females
– 11 males• 4 animals unknown
• Age– 20 adults
– 4 juveniles
– 1 fetus• Important implications for humans
and companion animal health
• Venereal or hematogenous mechanisms
Photo: Dr S Norman
Photo: Dr A Hall
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Pneumonia
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Lymphadenopathy
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Ancillary Laboratory Results
• Fungal culture
– Consistently isolated Cryptococcus gatti, type B from lung and mediastinal lymph nodes
– PCR-fingerprinting, RFLP
• Isolates identified as genotype VGII
• Only 2 isolates VGI
– Human fatality
– Dall’s porpoise from West Vancouver, mixed VG1/VG2
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Ancillary Laboratory Results• For each submission
– Pooled lung, lymph node, spleen, and brain• Negative for morbillivirus and Brucella spp by polymerase chain reaction
(PCR)
– Fecal floatation identified light to moderate burden of gastrointestinal nematodes
– Seven of 25 liver samples with no detectable vitamin A, 3 with moderately reduced values
– Retinyl palmitate and retinol
– Reference range 300-1200 mcg/gm dry weight
– Immunohistochemistry on select cases• Antibodies available for Cryptococcus spp capsular staining, but not as specific
as culture or molecular studies
• To date, no consistent pre-existing or underlying diseases detected within examined porpoises– Contaminant studies currently underway
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Porpoises at VAMMR 2008 to present
• 6 live-stranded
• 6 weeks to adult
– Bimodal stranding pattern
with peaks in early spring
and late summer
• 12.1 to 49.5 kg at admit
• Each animal received 24-
hour care by veterinary staff
and rescue volunteers.
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Theo
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Theo
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Theo
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Theo
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Theo
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Theo
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Theo
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Theo
• 3 year-old, 41 kg, male sub-adult live
stranded Pacific harbor porpoise (P.
phocoena) admitted July 2012
• Poor body condition, moderate lethargy,
marked weakness, and inability to swim
• moderate dehydration, mucopurulent
discharge from the blowhole, blepharospasm
OU and multiple superficial to deep
necrotizing cutaneous wounds
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Theo
• Elevated liver enzymes and mild inflammation
• Fecal parasitology identified Nasitrema sp. and
Stenurus sp. ova and larvae
• Differentials for the clinical signs included peri-
stranding trauma, septicemia, verminous pneumonia
and malnutrition
• Therapy consisted of antibiotics, corticosteroids,
benzodiazepines, antiemetics, bronchodilators,
fenbendazole, praziquantel, rehydration and wound
care
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Theo
• 30 days after admission, marked
inflammation was noted
• Yeast was found associated with skin wounds
as well as in gastric samples
• Itraconazole started and inflammation
resolved over the next 16 days and porpoise
appeared better
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Theo
• HOWEVER, 58 days after admission, the
porpoise began to slowly deteriorate showing
increased vomiting, weakness and decreased
mobility
• Developed marked scoliosis, head pressing
and erratic head movements
• Radiographs, sonography, and gastroscopy
revealed no significant lesions
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The Shanie Factor
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• High-field (1.5T) whole-body magnetic resonance imaging (MR)
was performed on Day 71
• MR revealed patchy, coalescing, solid intra-axial brain lesions in
the basal ganglia (caudate nucleus, putamen) and thalamus that
were hyperintense on T2w and FLAIR sequences
• Due to ongoing deterioration and poor prognosis, the porpoise was
euthanized
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Cryptococcus testing
• Latex agglutination for cryptococcal antigen
• Ante mortem at Idexx
– NEGATIVE
• Post mortem hindsight at human hospital
lab
– 1:8
– POSITIVE
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Conclusions
• First clinical presentations of a primarily neurologic form
of cryptococcosis in a cetacean
•
• First antemortem detection of imaging findings consistent
with central nervous system cryptococcosis
• These findings highlight the need for more rapid
diagnosis to provide better case management for stranded
cetaceans.
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And other marine mammals ?
• Since 2014, cryptococcosis has been diagnosed
in two live-stranded harbour seals admitted to the
VAMMR
• A female 3 week old pup and an unrelated adult
male
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Clinical presentation
• Pup was malnourished, dehydrated and had
multiple puncture wounds
– Significant leukocytosis
– Died 3 days after admit with mucohemorrhagic nasal
discharge
• Adult was malnourished and dehydrated and
poorly responsive
– Moderate leukocytosis
– Died on second day after admit during a prolonged
seizure
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Necropsy results
• Consistent in both animals
– Granulomatous lymphadenitis
– Menigoencephalitis
– Bronchopneumonia
– Urinary cystitis
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Histopathology
• Extracellular yeast consistent with
Cryptococcus spp.
– Lymph nodes
– Brain
– Lung
– Urinary bladder
• Peripheral fungemia
• Cryptococcomas in the pancreas, salivary
gland, trachea, and thyroid gland
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Localized meningoencephalitis
with focal effacement of the
molecular layer of the
cerebellum and peripheral
displacement and compression
of the adjoining granular layer
Cerebellar necrosis and
associated inflammatory
infiltrate and intralesional yeast.
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Lung with varying degrees of
atelectasis and congestion as well
as distended alveoli, occluded by
a myriad of yeast and abundant
mucus
Lymph node involvement with
expansion and replacement of the
cortex by nodular to diffuse
accumulations of yeast with mild
to moderate granulomatous
inflammatory infiltrate
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Additional diagnostics
• Genotyping studies confirmed isolates as C.
gattii VGIIa
• Serum antigen titres for Cryptococcus spp.
were > 1,024 fro both animals
– CrAg Lateral Flow Assay, Imprimis
Pharmaceuticals
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Acknowledgements
• Vancouver Aquarium’s Marine
Mammal Rescue Centre
• Shanie Fradette
• The marine mammal team
• The amazing vet techs: Chelsea
DeColle and Gwyneth Nordstrom
• The incredible rescue team: Kate
Cooper, Taryn Roberts, Sion
Cahoon
• > 80 volunteers and interns for
their invaluable efforts
• And a whole lot of support from
the whole Vancouver Aquarium
team