Procedure 1
Immerse in the stain for 3 minutes using a Coplin jar or beaker Do not allow aerosols to form during the process of drying slides Wayson Stain Visualization of Plague Bacilli Wayson stain is a polychromatic differential stain suitable for the detection of Yersinia and Pasteurella species. All Y. pestis organisms are Wayson-positive, but all Wayson-positive strains are not Y. pestis.
Gispen Silver Stain Visualization of Cytoplasmic Smallpox Virus Aggregates
Gispen's modified silver stain is in reality one of many existing modifications of Golgi's method based on the use of silver chromate as the staining medium. Although the use of the stain is quoted in most papers and books including the present one dealing with the detection of variola virus using laboratory methods, no references to the methodology are provided. For interested readers, the method is described in Geisert, E.E., Jr. and Updyke, B.V., Chemical stabilization of Golgi silver...
Results
Plague short, fat coccobacilli with bipolar safety pin staining patterns Smallpox Guarnieri bodies reddish cytoplasmic inclusions Phosphate buffer Sorensen - Stock A 0.2 M sodium dihydrogen orthophosphate NaHpOHO, MW156 dissolve 3.12 g in 100 ml distilled water - Stock B 0.2 M disodium hydrogen orthophosphate Na2HPO dissolve 2.83 g in 100 ml distilled water - Mix 25.5 ml of A with 13.3 ml B add distilled water to TOTAL 100 ml the resulting pH will be 6.8 - Mix 7.36 g Giemsa stain powder in 500...
Procedure
Cover slide with Lillie's crystal violet solution for 1 minute Hold the slide downward, rinse briefly in tap water Pour Gram's iodine, allowing it to flow downward along the surface of the slide for 2 minutes Differentiate in acetone 1 or 2 seconds by holding the slide downward and pouring the acetone over the slide from the upper end Wash thoroughly in running tap water, then check the stain under a microscope. The background should NOT be purple Counterstain 30 seconds with safranine Do not...
Sputum Smear Preparation
Place a clean microscope slide sample slide firmly on a bench Using a clean glass rod place a few droplets of the sputum on the sample slide and spread the droplets to produce an even film Fix the slide in absolute methanol for 3 minutes If a thin smear is wanted, use the procedure cited earlier for a thin blood smear Do not allow aerosols to form during the process of drying slides
Brucellosis
Brucellosis is a zoonotic disease of domestic and wild animals caused by bacteria belonging to the Brucella genus. Four species of these Gramnegative bacteria cause disease in humans. In aerosol form, the bacteria are highly infectious. Only 10 to 100 organisms are sufficient to cause disease in humans. The disease has a low 5 of untreated cases mortality rate, with deaths, typically due to endocarditis or meningitis. It may be incapacitating and even disabling. Frequently it becomes chronic...
Glanders And Melioidosis
These are zoonotic diseases. Glanders is caused by Burkholderia mallei and meliodiosis is caused by B. pseudomallei. Both are Gram-negative and present a safety-pin appearance. Infection in animals and humans is caused by inhalation or contamination of injuries. B. mallei induces disease in horses, mules, and donkeys. The rare instances of human infection are typically reported in veterinarians, horse and donkey handlers, and slaughterhouse workers. B. mallei is common in tropical and...