Culture media used for fungi Sabouraud Dextrose Agar (SDA)

Sabouraud dextrose agar (SDA) is a general-purpose medium used in microbiology and mycology labs for the isolation, growth, and maintenance of fungi. Most mold fungi do well on it, but it is especially recommended for growing dermatophytes (skin, hair, and nail fungi), yeasts, and other species found on animals and humans.

Constituents of SDA: the basic constituents of SDA are Water 4% to 7% dextrose: The high sugar concentration results in a high osmotic pressure, which tends
to inhibit the growth of bacteria 1% to 2% agar: acts as solidifying agent 1% peptone as nitrogen source Antibiotics such as penicillin, streptomycin, chloramphenicol, cyclohexamide can be used to inhibit the bacterial growth

Dermatophyte test medium (DTM)

Dermatophyte Test Medium (DTM) is a specialized agar used in medical mycology. It is based on Sabouraud's dextrose agar with added cycloheximide to inhibit saprotrophic growth, antibiotic to inhibit bacterial growth, and phenol red as a pH indicator. The pH indicator is useful in distinguishing a dermatophyte fungus, which utilizes nitrogenous material for preferred metabolism, producing alkaline by-products, imparting a red color change to the medium. Typical saprotrophic fungi utilized carbohydrates in the medium producing acidic by-products and no red color change.

Malt Extract Agar

Malt extract agar is used as a plate medium to isolate ringworm fungi. The medium is made selective by adding cycloheximide and chloramphenicol. Malt extract agar is most economically prepared from the dehydrated medium. Composition: Malt extract 30 gm/ltr Mycological peptone 5 gm/ltr Agar 1.5 gm/ltr

Inclusion body:
Inclusion bodies are nuclear or cytoplasmic aggregates of stainable substances, usually proteins. They typically represent sites of viral multiplication in a bacterium or eukaryotic cell and usually consist of viral capsid proteins. Examples of viral inclusion bodies in animals are Intracytoplasmic eosinophilic Negri bodies in Rabies Guarnieri bodies in Small pox Councilman bodies in hepatitis Negri bodies are eosinophilic, sharply outlined, pathognomonic inclusion bodies (210 m in diameter) found in the cytoplasm of certain nerve cells containing the virus of rabies, especially in Ammon's horn of the hippocampus. Often also found in the cerebellar cortex of postmortem brain samples of rabies victims. They consist of ribonuclear proteins produced by the virus.

Figure: Negri bodies, which are cellular inclusions found most frequently in the pyramidal cells of Ammon's horn, and the Purkinje cells of the cerebellum. They are also found in the cells of the medulla and various other ganglia.

Guarnieri bodies are cellular features found upon microscopic inspection of epithelial cells of individuals suspected of having poxvirus (e.g. smallpox or vaccinia). In cells stained with eosin, they appear as pink blobs in the cytoplasm of affected epithelial cells. The absence of Guarnieri bodies cannot be used as to rule out smallpox, however, as more sensitive test need to be performed. Councilman body is an eosinophilic globule often found in the liver of individuals suffering from viral hepatitis, yellow fever, or other viral syndrome. It represents a hepatocyte that is undergoing apoptosis (controlled cell death).

Councilman body (upper-right) and ballooning degeneration (centre-left).

Bacteriophage:
Bacteriophages are viruses that infect bacteria. They do this by injecting genetic material, which they carry enclosed in an outer protein capsid. The genetic material can be ssRNA, dsRNA, ssDNA, or dsDNA ('ss-' or 'ds-' prefix denotes single-strand or double-strand) along with either circular or linear arrangement. Examples of bacteriophages: Lambda phage ( phage); T2 phage; M13 phage, etc.

Neutralization test:
Neutralization of a virus is defined as the loss of infectivity through reaction of the virus with specific antibody. Virus and serum are mixed under appropriate condition and then inoculated into cell culture, eggs or animals. The presence of un-neutralized virus may be detected by reactions such as Cytopathic Effect (CPE), haemadsorption/haemagglutination, plaque formation, disease in animals. The loss of infectivity is bought about by interference by the bound Ab with any one o the steps leading to the release of the viral genome into the host cells. There are two types of neutralization;-