Fungi Allergy and


Some Topics

Dr. Gislason's 2-Slide Culture of Blastomycosis

Quick Identification of Fungal Infection in Sputum Samples

The diagnosis of fungal infections of the lung are difficult to make. The technique described here would allow a physician or lab tech to take a sputum sample from patient and within a few hours make the diagnosis:

I developed a nose, throat and lung infection in the summer of 2006. I realized that I had an infection that I did not recognize which would be difficult to diagnosis. I would cough up clumps of thick, white mucous. Sputum cultures were reported as negative and although I had many blood abnormalities, none pointed to a specific infection. Fortunately, I am a curious and have a lab with microscopy capabilities.

I discovered a simple culture technique: I added one or two drops of distilled water and spread the thick white mucous on a glass slide. This was covered with a second slide. The sputum specimen sandwiched between two glass slides becomes a fungal culture. In this preparation I observed that yeast cells would aggregate and metamorphose into a mycelia form at room temperature. All 2-slide cultures were stored at room temperature and examined at intervals.

Migration and aggregation of yeast cells

Within a few hours, the yeast cells clustered and formed tubes in the central portions of the culture. At the periphery of the slide, yeast cells would aggregate along the edges to form single walls that grew toward each other and formed tubes toward the center. All other cells in the mucous would be trapped and digested in these dammed areas. At 24 hours hyphae were developing.

The migration and aggregation of yeast cells was helpful in their identification since infected mucous contains a dense population of mixed cells. Also mucous from the trachea and bronchi contains a variable mixture of inhaled particles, pollens and spores. While you might see yeast cells in this dense aggregation, there became more obvious as they migrated away form the other cells and clustered.

To view the fresh, alive specimen, no staining is required. The microscope condenser should be low and the diaphragm closed to enhance contrast. I examine the slide first with low power lens (40X) and then at 160 X. The specimen then remains alive and has a depth of field. Here is an example of the 2-slide culture after 4 days at room temperature. A branching root-like growth is obvious. This, of course, is not a traditional fungal colony that grows in 3 dimensions on an agar medium, but a novel two-dimension growth pattern that occurs in a natural culture medium (sputum). In the longest-lived cultures, tangled mycelial growth appears and conidiophores sometimes develop days later. Further growth irregularities were probably caused by itraconazole which I was taking when the samples were collected.

After hours to days, I would divide some slides and examine them under higher power. I would stain some slides and examine others without staining. The following photomicrographs were of unstained specimens. Under the microscope the early stages (4hours) of yeast cell aggregation and transformation, looked like this:

Blastomycosis yeast cells transforming into mycelial form... hyphus creation at 6 hours.The yeast cells, elongate, flatten and eventually disappear, leaving behind relatively cellular chitin tubes that grow outward and branch. 2-slide culture image

Closeup, contrast enhanced image

At 36 hours

1000x image of hyphus after 36 hours. Note double walls and and open end. It appears that digestive chemicals and enzymes are secreted by the tubes, digesting material from the adjacent area.As the hyphae develop, the slide is cleared of mucous and other cells.

See More Images

Fungi Everywhere

A profusion of fungi exists in the environment. Some fungi are able to cause an invasive infection in otherwise healthy individuals. Other fungi are opportunistic fungi that become invasive when immune defenses are compromised. Diagnosis of fungal infection is difficult. There are many problems when you try to connect a test result to a disease. Fungi are so abundant and there are so many varieties in every environment that it is seldom easy to pick just one cause among many. Fungi are inhaled and ingested. Foods always contain fungal spores and actively growing molds. Attempts to culture fungi often fail; only a small number grow in the culture media commonly used. Some new methods of detecting fungal DNA may be useful but development of reliable tests is slow and expensive.

'You are at Alpha Online, a service of Environmed Research Inc. Sechelt, British Columbia Canada. Orders for printed books, eBooks for download and nutrient formulas are placed at Alpha Online.

 Secure Order Site    Secured by GeoTrust QuickSSL

Order Alpha Education Books

 The books in this series address important medical problems and their solution. Everyone, who is interested in Nutritional Therapy, will need a copy of the Alpha Nutrition Program. All books can be downloaded as eBooks (PDF files).

Click the add to cart buttons to begin your order for printed books for mail delivery to the USA and Canada. Click the Download buttons to order eBooks for download.

Print Books Read Topics Download
Alpha Nutrition Program
Aching and Fatigue
Air and Breathing
Alpha Nutrition Cooking
Alcohol Problems 
Gluten Problems 
Diabetes 2
Eating & Weight 
Skin Disease
Feeding Children
Human Brain
Food Allergy
Food & Digestion
Food Choices
Heart & Arterial
Immunology Notes
Inflammatory Arthritis
Nutrition Notes

alpha online

We ship through the Post Office to all destinations in Canada and the USA. All correspondence about your order is done by email- send to Alpha Mail. All orders are completed with encrypted security. Payments are made through Pay Pal by account or credit cards.

Persona Digital publishes a series of books on current topics in psychology, sociology, neuroscience and philosophy. eBooks and can be downloaded to any destination on the planet. Printed books and eBooks are ordered from Alpha Online. Printed books are shipped only to Canada and the USA by postal services.

Alpha Nutrition is a trademark and a division of Environmed Research Inc., founded in 1984 at Vancouver, BC, Canada. Online Since 1995. Experts in Self-Managed Care. Experts in Elemental Nutrient Formulas. We are located on the Sunshine Coast in Sechelt, a ferry ride from West Vancouver.