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MTEC 216 Assignment (Midterm) A. Developmental Organelles Inclusions 1.

Howell Jolly Bodies Description - small, round fragments of the nucleus (resulting from karyorrhexis or nuclear disintegration) of a late nucleated red cell or metarubricyte - stain reddish-blue with Wright stain - may also result from incomplete extrusion of the nucleus or from chromosomes that were separated from the spindle during abnormal mitotic division

2. Basophilic Stippling

- fine or coarse, deep blue to purple staining inclusion that appears in erythrocytes on a dried Wright-stained film. - much smaller than Howel-Jolly bodies - usually irregularly shaped - appear homogenously throughout the Hb portion of the erythrocyte represents aggregates of ribosomes that appear during the drying and staining of films.

3. Pappenheimer Bodies - also called siderotic granules - small, irregular, dark-staining granules that appear near the periphery of a young erythrocyte in a film stained with Wright or supravital stain. - with Perls Prussian blue stain, these bodies stain positively 4. Polychromatophilic Red cells - aka diffusely basophilic red cells - young red cells that no longer have a nucleus but still contain some RNA - as the name implies, this RNA stains diffusely blue with Wright stain - slightly larger cells than mature red cells - contain ribosomes, mitochondria, and other granules

5. Cabot Rings

- thin ringlike structures - stain reddish violet in Wright stain - may be circular or appear at the cell periphery, - may form a figure of eight, incomplete rings, or other configurations - may be observed in a nucleated red cell or be associated with stippling or a HowellJolly body in the same erythrocyte

B. Abnormal Hemoglobin Precipitation Inclusions 1. Heinz Bodies Description - round, refractile inclusions not visible on a Wrightstained film - range in size from about 1 to 3 m - attached to the erythrocyte membrane - best identified by supravital staining with basic dyes such as crystal violet, methylene blue, or brillian cresyl blue 2. Hemoglobin H Inclusions - small greenish-blue inclusion bodies appearing in erythrocytes after 4 drops of blood is incubated with 0.5 mL of 1% brilliant cresyl blue for 20 minutes at 37 0C. - represent precipitated Hb H - can also be seen in early nucleated red cells in the bone marrow of patients with Hb H disease.

C. Protozoan Inclusions Plasmodium vivax Malaria Inclusions 1. Ring stage

Appearance of RBC -normal to 1.25x, round; occasionally fine Schffners dots; multiple infection of RBC not uncommon

Appearance of parasite -large cytoplasm with occasional pseudopods; large chromatin dot

2. Trophozoite -enlarged 1.5 to 2x; may be distorted; fine Schuffners dots -large amoeboid cytoplasm; large chromatin; fine yellowish-brown pigment

3. Schizont -enlarged 1.5 to 2x; may be distorted; fine Schffners dots -large, may almost fill RBC; mature = 12 to 24 merozoites; yellowish-brown, coalesced pigment

4. Gametocyte -Enlarged 1.5 to 2x; may be distorted; fine Schuffners dots -Round to oval; compact; may almost fill RBC; chromatinb compact, eccentric (macrogametocyte) or diffuse (microgametocyte); scattered brown pigment

Plasmodium ovale Malaria Inclusions 1. Ring Stage

Appearance of RBC -normal to 1-1/4 X, round to oval; occasionally Schffners dots; occasionally fimbriated; multiple infection of RBC not uncommon

Appearance of parasite -sturdy cytoplasm; large chromatin

2. Trophozoite -normal to 1-1/4 X; round to oval; some fimbriated; Schffners dots -compact with large chromatin; dark-brown pigment

3. Schizont -normal to 1-1/4 X; round to oval; some fimbriated; Schffners dots -mature = 6-14 merozoites with large nuclei, clustered around mass of dark-brown pigment

4. Gametocyte -normal to 1-1/4 X; round to oval; some fimbriated; Schffners dots -round to oval; compact; may almost fill RBC; chromatin compact, eccentric (macrogametocyte) or more diffuse (micro- gametocyte); scattered brown pigment

Plasmodium malariae Malaria Inclusion 1. Ring stage

Description -P. malariae rings have sturdy cytoplasm and a large chromatin dot

2. Trophozoite -The parasite is smaller than red blood cells and contains 6-12 merozoites, usually 8 in a single ring. The pigment is a brown / greenish central clump. The red cell slightly enlarged, stippled, frequently oval and fimbriated.

3. Schizont -A schizont containing merozoites (6 to 12) giving a coarse granular appearance.

4. Gametocyte -The gametocytes of P. malariae have a round shape about the size of red blood cells. They have a fine granular appearance. -Gametocytes contain large amounts of black pigment, with chromatin present as a compact mass in females and diffuse in males. They occupy less than two thirds of the red blood cell.

Plasmodium falciparum Malaria stage 1. Ring stage

Description -P. falciparum rings have delicate cytoplasm and one or two small chromatin dots. Rbcs that are infected are not enlarged; multiple infection of rbcs is more common in P. falciparum than in other species. Occasional appliqu forms (rings appearing on the periphery of the rbc) can be present.

2. Trophozoite -The cytoplasm of mature trophozoites tends to be more dense than in younger rings. As P. falciparum trophozoites grow and mature, t they tend to retain their ring-like shape and sometimes trace amounts of yellow pigment can be seen within the cytoplasm. Growing trophozoites in P. falciparum can appear slightly amoeboid in shape.

3. Schizont -P. falciparum schizonts are seldom seen in peripheral blood. Mature schizonts have 8 to 24 small merozoites; dark pigment, clumped in one mass.

4. Gametocyte - P. falciparum gametocytes are crescent or sausage shaped. The chromatin is in a single mass (macrogamete) or diffuse (microgamete).

References: CDC (n.d.) Retrieved from http://www.cdc.gov/dpdx/resources/pdf/benchAids/malaria/Pfalciparum_benchaidV2.pdf on January 19, 2014 CDC (n.d.) Retrieved from http://www.cdc.gov/dpdx/malaria/dx.html on January 19, 2014 Malaria (n.d.) Retrieved from http://www.phsource.us/PH/PARA/Chapter_9.htm on January 19, 2014 Microscopy Plasmodium species (1999) Retrieved from https://www.msu.edu/course/zol/316/psppscope.htm on January 19, 2014 Parasitology Illustrated (2011) Retrieved from http://parasitologyillustrated.com/classes_of_parasites/protozoa/sporozoa/p_ovale.html on January 16, 2013 Red Blood Cell Inclusions (2009) Retrieved from http://hemeplace.wikispaces.com/file/view/RBC_Inclusions.pdf on January 18, 2014 CDC (n.d.) Retrieved from http://www.cdc.gov/dpdx/resources/pdf/benchAids/malaria/Pfalciparum_benchaidV2.pdf on January 19, 2014