MICROBIOLOGY SEM 5 2010/2011 PART II

1. A one-year-old boy with microcephaly, jaundice, hepatosplenomegaly and mental retardation. The pediatrician explains to the mother that Cytomegalic Inclusion Disease may be a provisional diagnosis. The virus being transmitted transplacentally. a) Laboratory procedures undertaken to get to the final diagnosis : (pg.102; part 2) i. CMV antigen detection in peripheral blood mononuclear cells by indirect immunofluorescent technique ii. CMV DNA detection by qualitative PCR and assessment of the viral load by quantitative PCR iii. Growth in vitro on TCC : CMV is permissive in human fibroblasts. Shell vial assay which is a vial-coated with human fibroblast monolayer that could be inoculated with the peripheral blood leukocytes, has overcome the high costs and the time consumption of the conventional TCC. iv. Indirect detection which includes the serologic determination of CMV IgM & IgG. 2. A 35-year-old female suffering from a painless swelling around the index nail finger of her right hand, oozing pus on pressure, the proximal part of the nail was dystrophic & yellow in color. Her dermatologist advised her to perform direct KOH examination & fungal culture. a) The previous clinical condition is : (pg.13; part 2) Onychomycosis b) The most propable etiologic agent : (pg.13; part 2) Trichophyton species (T.rubrum, T.mentagrophytes, T. tonsurans) c) Direct KOH microscopic picture : (pg.14; part 2) Branching hyphae, chains of arthropores or a mixture of both d) Culture medium used for isolation : (pg.15; part 2) Sabourauds Dextrose Agar (SDA) e) The most suitable antifungal agent to be prescribed : (pg.13; part 2) Griseofulvin

3. Choose the correct method of sampling for each of the following FUNGALLY infected sites. (write down the method for the infected site) (pg.5-6; part 2) a) Infected nails Scrapings are taken from the deeper parts b) Mucous membrane infection Dry swabs c) Bronchopulmonary infection Morning sputum d) Urinary tract infection Mid-stream urine e) Cerebrospinal infection Collect CSF into sterile test tubes

PART III
1. Tinea capitis : Pathogenesis (pg.12-13; part 2) (a) Gray-patch ringworm :  The fungus primarily colonizes the outer portion of hair shafts (ectothrix hair involvement)  The lesions are seldom inflamed, but luster and colour of the hair saft may be lost (b) Black-dot ringworm :  Hair follicle is the initial site of infection  Fungal growth continues within the hair shaft causing it to weaken (endothrix hair involvement)  The brittle, infected hair shafts break off beneath the scalp leaving the black-dot stubs Laboratory diagnosis (pg.14-15; part 2) (a) Specimen :  Cleanse the affected area with 70% ethanol  Collect by removing dull broken hairs from the margin of the lesion using sterile tweezers (b) Direct microscopy :  Place a drop of potassium hydroxide solution.  Transfer the hair (small pieces) to the drop of KOH and cover with a cover glass. Clearing can be hastened by gently heating the preparation over the flame of Bunsen burner.  Examine it microscopically using the 10x and 40x objectives.  Look for arthrospores and hyphae, and note whether the infection is on the outside of hair (ectothrix) or within it (endothrix).

(c) Culture :  Culture the fungus on SDA which contains Cycloheximide (Actidione) to get rid of saprophytic fungi and an antibiotic such as chloramphenicol  Incubate aerobically at room temperature (25-30oC) for up to 3 weeks, examining every few days for growth  Identify hyphal characteristics, absence or presence of spores and morphology of the spores 2. Tetanus is sometimes referred to as lock jaw. a) Explain how infection with this organism can end with this picture. (pg. 124; part 1)  Toxin tetanospasmin reaches the CNS by retrograde axonal transport or via the blood stream.  It becomes fixed to ganglioside receptors in the brain stem and spinal cord.  It inhibits the release of inhibitory mediators (glycine & GABA) at the terminals of inhibitory neurons resulting in the excessive activity of excitatory motor neurons.  This results in muscle spasm and hyperreflexia.  Muscular spasms often involve first the area of injury and infection and then the muscles of the jaw (lock jaw) b) Tetanus is a totally preventable disease. Explain how. (pg. 125; part 1)  Active immunization with toxoids  Proper care of wounds contaminated with soil  Prophylactic use of antitoxin  Administration of penicillin 3. Features of Small pox that made it eradicable. (pg. 92; part 2)  Stable serotype  Effective vaccine  Long-term immunity conferred by infection or vaccination  No animal reservoir  Easily diagnosed  No latency, no carrier state, no subclinical or persistent infections