Prepared By: Ken Robin A. Canada, RMT, Mls Ascpi
Prepared By: Ken Robin A. Canada, RMT, Mls Ascpi
Prepared By: Ken Robin A. Canada, RMT, Mls Ascpi
During the rainy season, the mycelia grow rapidly but they are also the least
infectious form of the organism.
Arthrospores measure 3-5 µm and are extremely hardy, withstanding extreme heat,
desiccation, and changes in soil salinity and remaining viable in the soil for months to
years.
Upon inhalation into lungs or on rare occasion after percutaneous implantation into
tissue, each arthroconidium transforms into a new multinucleated, spherical
structure called the spherule.
As it grows, the spherule begins to divide internally. Spherules vary from 60 to over
100 um in diameter, while endospores remain 2 to 5 um in size.
Mature spherules may contain 800 to 1,000 endospores.
In its parasitic phase, each endospore grows into a new spherule.
Coccidioides spp. are found in the hot, dry regions of the southwestern United
States, where winters are relatively mild and the soil is alkaline.
Coccidioides spp. are most highly concentrated in the San Joaquin Valley of CA and
in south-central AZ.
Historically, people at greatest risk for contact include farmers, construction
workers, and archaeologists.
The spectrum of illness due to Coccidioides spp. is very broad.
The 40% of patients that are symptomatic may present with an acute or subacute
spectrum of illness, ranging from “flu-like” to progressive pneumonia.
The incubation period of coccidioidomycosis usually begin within 7 to 21 days of
inhalation of arthroconidia.
Symptomatic patients complain of fever, cough, chest discomfort, malaise, and
fatigue (generally last less than 3 weeks)
Transient skin manifestations, including rash and erythema nodosum, may be seen in
10% to 50% of patients.
Patients of black or Asian (especially Filipino) ethnic backgrounds, pregnant women
in the third trimester, and any immunocompromised patients appear to be at
significant risk for disseminated coccidioidomycosis.
Dissemination may occur months to several years after the primary infection.
Dissemination of spherules and endospores via the lymphatics and the bloodstream
may occur to any organ system, but skin, lymph nodes, and the skeletal system are
primarily involved.
Hematology.
Elevated erythrocyte sedimentation rate and eosinophilia may be seen in
coccidioidomycosis. Eosinophilia especially should heighten suspicion.
Direct detection.
(i) Microscopy
This includes a mixed acute reaction consisting of an influx of polymorphonuclear
neutrophils and, to a far lesser degree, granulomatous cells.
Tissue eosinophilia may be present and may be seen surrounding the offending organism
(Splendore-Hoeppli phenomenon).
Granuloma Formation Splendore-Hoeppli phenomenon
Coccidioidomycosis can be diagnosed microscopically by visualization of endospore-
containing spherules in infected material.
Potassium hydroxide (KOH) wet mounts are useful and readily available for
microscopic evaluations
Calcofluor white (CFW) fluorescent stain
Nucleic acid amplification tests using PCR have been described by a number of
noncommercial laboratories and found to be both sensitive and specific.
For growth in the spherule phase, arthroconidia were harvested from 4-week-old
cultures. Inoculated into 1 liter modified Converse medium (7) supplemented with
0.05% NZ-Amine and cultures were incubated at 39°C, 8% CO2, and 160 rpm, for
96h.
Mycelia for protein extraction were grown in liquid 2XGYE at 37°C and 180 rpm, for
48 h.
Spherules were harvested by centrifugation and the carbohydrate rich outer wall
material removed from the surface of the cell pellet.
The cell pellet was washed twice with water, then frozen at 80°C.
Mycelia were harvested by filtration, washed extensively with water, and frozen at
80°C.
The protein concentration for each extract was determined using the 2-D Quant kit.