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Parasitology

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Specimen Collection and Handling : general   • 3 specimens w/in 10 day every other day • Liquid within 30mins of passage or place in preservative • Soft specimens w/in 30mins of passage or place in preservative • Fformed stools w/in 3-4hrs; place an aliquot in preservative&frige remainder  
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Types of Specimens   • Feces- 95% parasite specimens • Sputum- early morning specimen is best (most concentrated) • Urine- early morning or 1st void • Genitalia- saline wet swabs • Tissue and skin- sterile container • Blood  
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Types of Specimens: feces part 1   • Collect in clean, dry container with secure lid  
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Types of Specimens: feces part 2   • Do not accept specimens contaminated with urine (may destroy motile organisms), water (may contain free living organisms), or oil or barium enemas (intestinal protozoa may be undetectable 5-10 days after barium is given)  
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Types of Specimens: feces part 3   • Antibiotics (such as tetracycline) modify intestinal flora and may prevent parasite recovery for 2 weeks after drug cessation  
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Types of Specimens: blood   • Fresh blood from finger stick (best NOT to use EDTA) • Prepare thick smear for concentration and thin smear for identification  
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Ova, Cyst, and parasite examination: macroscopic part 1   • Consistency: liquid-trophozoites soft-protozoan cysts and trophozoties formed-protozoan cysts (heiminth eggs can be found in any type of stool)  
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Ova, Cyst, and parasite examination: macroscopic part 2   • Presence of: mucus, blood or macroscopic parasites • Color: black-blood, clay colored-barium or absence of bile, green-veggies  
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Ova, Cyst, and parasite examination: microscopic   • Heavy amounts of yeast (yeast may be the cause of diarrhea) • Increased WBCs (may indicate a bacterial cause of diarrhea) • All Ova, Cysts, and Parasites  
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Ova, Cyst, and parasite examination: permanent smears   • Iron hematoxylin stain- takes skill to perform; organisms stain shades of purple-gray • Trichrome stain- easy to perform; organisms stain great and nuclear details stain red-black • Lawless stain- rapid, 5mins stain; stains various shades of blue  
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Types of Preservation   • Refrigeration • 10% formalin • MIF (Merthiolate-Iodine-Formalin) • PVA (Palyvinyl Alcohol) • SAF (Sodium Acetate-Acetic Acid-Formalin)  
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Types of Preservation : Refrigeration   Good for eggs, larvae, and amoebic cysts; DO NOT refrigerate if you  
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Types of Preservation : 10% formalin   Good for eggs, larvae and amoebic cysts  
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Types of Preservation : MIF (Merthiolate-Iodine-Formalin)   Concentration procedures as well as preservative; good for eggs and amoebic cysts  
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Types of Preservation : PVA (Palyvinyl Alcohol)   Best for amoebic trophozoites; can prepare permanent stain slides from specimens preserved this way  
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Types of Preservation : SAF (Sodium Acetate-Acetic Acid-Formalin)   Good for amoebic trophozoites; environmentally safer than PVA  
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Concentration techniques: (used to detect small number of parasites)   • Formalin-Ether Technique- Modified version replaces ether with ethyl-acetate (less volatile) • Zinc Sulfate Flotation Technique • MIF  
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Concentration techniques: Formalin-Ether Technique part 1   • Forms 4 layers: Ether (ethyl acetate) Debris Formalin Sediment (parasites); • Ether removes fats and oils and formalin preserves organisms  
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Concentration techniques: Formalin-Ether Technique part 2   • Advantages: can stay in formalin stage indefinitely; easy to perform • Disadvantages: small ova and cysts such as Giardia end E. nana do not sediment well  
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Concentration techniques: Zinc Sulfate Flotation Technique part 1   • Specific gravity of zinc sulfate is greater than ova, cysts, and larvae, therefore they float on top of zinc sulfate solution • Specific gravity of zinc sulfate solution; if specimen is in saline =1.18. if specimen is in formalin =1.20  
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Concentration techniques: Zinc Sulfate Flotation Technique part 2   • Advantages- easy to perform; does not use flammable chemicals • Disadvantages- distorts some helminth eggs; large eggs (ex. Schistosomes) and operculated eggs (ex. D. latum) may sink quickly  
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Concentration techniques: MIF part 1   • Concentration and preservative • After adding stool to the vial, break iodine ampule (mix vial well). Allow to sit undisturbed for at least 2hrs • Forms 3 layers: Top (formalin) Interface (parasites) Bottom (fecal debris)  
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Concentration techniques: MIF part 2   • Advantages- easy to use; can inoculate on nursing floor • Disadvantages- merthiolate distorts some parasites; does NOT preserve trophozoites  
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Helminths :Intermediate host   • Host which contains the larval form of the parasite  
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Helminths: Definitive host   • Host which contains the adult sexual form of the parasite  
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Helminths: Hermaphroditic   • Contain both sexes in one helminth; cestodes and trematodes (except Schistosomes)  
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Helminths: Gravid proglottid   • Segments filled with eggs  
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Intestinal Nematodes (Roundworms): general characteristics part 1   • Females larger &have straight tail • Males curved tail with a spicule used for copulation • Unsegmented  
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Intestinal Nematodes (Roundworms): general characteristics part 2   • Round, elongated from a few millimeters-meter in length • Complete digestive tract (mount to anus) • Found in intestine, blood, and tissue • Worldwide distribution  
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Ascaris lumbricoides (Large intestinal Roundworm) part 1   • Largest nematode- adult males 150-200mm. Adult females 200-300mm • If no male present in small intestine; female will lay bizarre shaped unfertilized eggs • No intermediate host needed  
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Ascaris lumbricoides (Large intestinal Roundworm) part 2   • Second most common nematode infection in U.S. 5-9 year old group most revalent in U.S. • Visceral Larval Migrans- dog and cat ascarid (Toxocara canis and cati) migration through viscera resulting in eonsinophilia  
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Ascaris lumbricoides (Large intestinal Roundworm) Diagnosis   • Demonstrated characteristic egg in feces • Recovery of worms from anus, nasal passage, or sputum  
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Ascaris lumbricoides (Large intestinal Roundworm) Clinical disease   • “worm ball” –blockage in intestines • “ascaris pneumonitis” –due to larvae migration in lungs  
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Enterobius vermicularis (Pinworm) part1   • Adults Males 2-5mm Females 8-13mm • Eggs become embryonated within hours resulting in high rate of autoinfection; treat entire family of an infected individual  
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Enterobius vermicularis (Pinworm) part 2   • Most common helminth parasite of humans; frequently in children due to poor sanitation habits  
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Enterobius vermicularis (Pinworm) Diagnosis part 1   • Scotch tape (cellulose tape) preparation- egg and larvae stick to tape. Since migration of female occurs at night the prep is performed after patient has been sleeping or early in the morning  
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Enterobius vermicularis (Pinworm) Diagnosis part 2   • Adults may become “stuck” to the outside of the stool as it passes the perianal folds where the female migrates to lay eggs  
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Trichuris trichiura (Whipworm)   • Adults Males 30-45mm long, long curved tail prominent spicule Females 30-50mm long • Prolapsed rectum- may occur in heavy infections • Diagnosis- demonstrate characteristic egg in feces  
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Strongyloides stercoralis (Threadworm)   • Adults Smallest nematode Possess a long cylindrical esophagus and lack a posterior bulb • Eggs hatch in mucosa of intestine and are rarely seen in feces  
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Strongyloides stercoralis (Threadworm): Rhabditiform larva (noninfective)   • 250um long • Muscular esophagus (club-shaped anterior, a restriction, and a posterior bulb) • Short buccal cavity (approximately 1/3-1/2 width of body) • Prominent primordial genitalia  
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Strongyloides stercoralis (Threadworm): Filariform larva (infective)   • 600um long • Lacks a bulb on the esophagus • Notch at the end of tail  
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Strongyloides stercoralis (Threadworm): free living cycle part 1   • Male and female adults mate and the female lay embryonated eggs • Eggs hatch in the soil to release the rhabditiform larvae  
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Strongyloides stercoralis (Threadworm): free living cycle part 2   • If favorable environmental conditions exist the larvae will mature into a free-living adult • If unfavorable environmental conditions exist the larvae moult into the infective filiariform larvae  
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Strongyloides stercoralis (Threadworm): parasitic cycle part 1   • Filariform larvae penetrate the skin of the host • Larvae enter the circulatory system and migrate to the heart and then to lungs  
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Strongyloides stercoralis (Threadworm): parasitic cycle part 2   • In the lungs the larvae exit through alveoli, travel up the trachea, are swallowed and mature in the small intestines • The adults mate and the female burrows into the intestinal mucosa where she lays her eggs  
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Strongyloides stercoralis (Threadworm): parasitic cycle part 3   • Rhabditiform larvae hatch from the egg and then enter the lumen of the small intestines where they are passed out with the feces where they enter the free living cycle  
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Strongyloides stercoralis (Threadworm): autoinfection   Some of the rhabditiform larvae develop into filariform larvae in the bowel and re-infect the host  
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Strongyloides stercoralis (Threadworm): Clinical disease part 1   • 3 stages Cutaneous- initial skin penetration Pulmonary- larval migration through lungs Intestinal- symptoms depend on worm load. Immunocompromised patients may exhibit leukocytosis and eosinophilia  
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Strongyloides stercoralis (Threadworm): Clinical disease part 2   • Hyper-infection syndrome- may lead to death from tissue damage Occurs in the immunocompromised (ADIS, drugs) Can be transferred through organ transplantation  
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Necator americanus (New World Hookworm) and Ancylostoma duodenale (Old world Hookworm): Adults   • Male 8-11mm long; possess a bursa which aids in speciation of hookworm • Females 10-13mm long • Adults may live 2-14 years • Rarely seen in stools since firmly attached to mucosa  
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Necator americanus (New World Hookworm) and Ancylostoma duodenale (Old world Hookworm): Rhabditiform larvae   • 250um long • Bulbous esophagus • Long buccal cavity (approximately as long as width of body)  
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Necator americanus (New World Hookworm) and Ancylostoma duodenale (Old world Hookworm): Infective stage- filariform larvae   • 700um long • Straight esophagus (1/4 length body) • Pointed tail • Long buccal cavity  
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Necator americanus (New World Hookworm) and Ancylostoma duodenale (Old world Hookworm): Clinical disease   • Pneumonitis • Allergic reactions- “ground itch” • Anemia- each adult worm consumes 0.2ml of blood/day • Cutaneous larvae migrans- migration of dog hookworms (Ancyclostoma braziliensis and caninum) through subcutaneous tissue causing intense itching  
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Necator americanus (New World Hookworm) and Ancylostoma duodenale (Old world Hookworm): Diagnosis   • Demonstrate characteristic egg in feces • Do not see larvae in feces (unless specimen left at room temp)  
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Remember: differentiating hookworm adults   • NeCator- Cutting plates • AncylosToma- Teeth or DUOdenale:2 pairs  
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Blood and Tissue Nematodes: Trichinella spiralis (Trichina Worm) adults   • Males 1.5mm long, no copulatory spicule • Females 3mm long. Females bear larvae NOT eggs  
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Trichinella spiralis (Trichina Worm): Infective stage   Ingestion of encysted larvae in under cooked pork  
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Trichinella spiralis (Trichina Worm): Diagnostic stage   Muscle biopsy showing encysted larvae in striated muscle (larvae form pork migrate to skeletal muscle and penetrate individual fibers and begin to grow, eventually becoming encysted by infiltrating leukocytes)  
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Trichinella spiralis (Trichina Worm): Clinical disease   • Destruction of muscle cell • High eosinophilia (may reach 90%) • May have myocardial involvement  
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The Microfilariae general characteristics part 1   • Required an arthropod as an intermediate host. When infected arthropod takes a blood meal, the microfilariae are released into human host  
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The Microfilariae general characteristics part 2   • Diagnosis made by examining Giemsa stained think and thin smears (except Onchocerca volvulus- skin scraping from modules)  
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Wuchereria bancrofti: Microfilaria   • Sheathed • NO nuclei in tip of tail • Nocturnal periodicity- 9pm-2am greatest concentration in blood  
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Wuchereria bancrofti: Elephantiasis   Permanent blockage of lymphatic system can occur in the upper extremities  
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Wuchereria bancrofti: Diagnosis   Demonstrate in blood smears (thick and thin) sheathed microfilaria with no nuclei in tip of tail  
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Brugia malayi: Microfilaria   • Sheathed • Nuclei stop then there is a clear space with 2 distinct nuclei at tip of tail • Nocturnal periodicity  
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Brugia malayi: Elephantiasis   Restricted to the lower extremities  
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Brugia malayi: Diagnosis   Demonstrate in blood smear (thick and thin) sheathed microfilaria with 2 terminal nuclei in tail  
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Brugia malayi   Found in Far East  
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Loa Loa (eye Worm)   Found in Africa  
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Loa Loa (eye Worm): Microfilaria   • Sheathed • Nuclei to the tip of tail • Diurnal periodicity (microfilaria shed in peripheral blood constantly day or night)  
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Loa Loa (eye Worm): Causes   Causes calabar swellings (allergic reaction to warm migration in tissue and death in capillaries)  
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Loa Loa (eye Worm): Diagnosis   • Demonstrate in blood smear (thick and thin) sheathed microfilaria with continuous nuclei in tail • Worm may migrate across conjunctiva  
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Onchocerca volvulus (Blinding worm): Microfilaria   • Only pathogenic microfilaria which is NOT sheathed • NO nuclei in tip of tail • Found in nodules under skin, NOT in peripheral blood  
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Onchocerca volvulus (Blinding worm): Clinical disease   • Sever dermatitis (50-70% eosinophilia) • Microfilariae in ocular structures may result in blindness. Leading cause of blindness in Africa  
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Onchocerca volvulus (Blinding worm): Diagnosis   • Demonstrate from skin “snips”/tissue scrapings unsheathed microfilaria with NO nuclei in tail  
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Remember: ONchochercus Volvulus   Only pathogenic microfilariae with no sheath, no nuclei in tip of tail (ON is NO backwards)  
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Remember: Wucheriria BaNcrofti   NO nuclei in tip of tail  
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Remember: Brugia Malyi   B is second letter in alphabet (2 nuclei in tip of tail)  
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Remember: Loa Loa   Name repeats and so do nuclei, continuously in tip of tail  
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Cestodes (Tapeworms) general part 1   • Flat, ribbon-like, segmented worms • Shape of proglottids (segments), presence or absence of armed rostellum (knob-like protrusion at the extreme anterior end of a tapeworm) and size aid in identification of adults  
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Cestodes (Tapeworms) general part 2   • Hermaphroditic- mature proglottids contain both male and female reproductive organs • 4 cup shaped suckers on scolex (except Diphyllobothrium latum which has 2 suctorial grooves) • Worldwide distribution  
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Taenia saginata (Beef Tapeworm)   Human is definitive host  
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Taenia saginata (Beef Tapeworm): adults   • Scolex has an unarmed rostellum • Consists of as many as 2,000 proglottids (10-15ft long) • Adults may live 25 years  
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Taenia saginata (Beef Tapeworm): Infective stage   Ingestion of under cooked beef containing larval stage  
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Taenia saginata (Beef Tapeworm): Diagnostic stage   • Find characteristic egg in feces • Proglottids can be stained; note number of major uterine branches (15-30 in T. saginata)  
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Taenia solium (Pork Tapeworm)   Human may serve as definitive or intermediate host  
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Taenia solium (Pork Tapeworm): adults   • Scolex has armed rostellum • Consists of any many as 1,00 proglottids; 6-10ft long  
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Taenia solium (Pork Tapeworm): Cysticercosis part 1   • Infection caused by larvae of parasite- human is intermediate host Man ingest the egg of T. solium Egg passes through the stomach and hatches in the intestine  
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Taenia solium (Pork Tapeworm): Cysticercosis part 2   • The embryo penetrates the mucosa and becomes a cysticercus. Most commonly found in the subcutaneous connective tissue, eye, brain, muscle, heart, and lungs • Cysticercosis- inflammatory response to tissue invasion and parasite death  
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Taenia solium (Pork Tapeworm): Diagnostic stage   • Find characteristic egg in feces • Proglottids can be stained; note number of major uterine brances (7-13 in T. solium)  
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Remember the hermaphroditic tapeworm   T: taenia species A: adult- small intestine P: proglottids contains male & female parts E: eggs-6 hooked embryos (hexacanth) W: worldwide nO: respiratory, vascular, digestive system R: ribbon like body. flat M: middle (intermediate) host require  
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Hymenolepis nana (Dwarf Tapeworm)   • Does not require an intermediate host; may have intermediate host such as flea or beetle • Most common human tapeworm in the U.S • Heavy infections can occur through autoinfection  
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Hymenolepis nana (Dwarf Tapeworm): adults   • Small tapeworm; 40mm long • Contains 200 segments • Armed rostellum (short with hooks)  
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Hymenolepis nana (Dwarf Tapeworm): stages   • Infective stage- ingestion of eggs • Diagnostic stage- demonstrate characteristic egg in feces  
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Hymenolepis diminuta (Rat Tapeworm)   • Adult- unarmed rostellum • Man is accidental host via ingestion of infected intermediate arthropod host (ex. Grain beetles in pre-cooked cereals) • Rat is definitive host  
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Hymenolepis diminuta (Rat Tapeworm): stages   • Infective stage- ingestion of flea • Diagnostic stage- demonstrate characteristic egg in feces  
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Diphyllobothrium latum (Broad Fish Tapeworm)   • Only cestodes to produce operculated eggs • High incidence in Finland, Alaska, and Canada  
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Diphyllobothrium latum (Broad Fish Tapeworm) : adults   • Scolex consists of 2 longitudinal sectorial grooves known as bothria, giving it a spoon shape • Uterus in gravid proglottids appears as a rosette • May have an many as 3,000 proglottids  
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Diphyllobothrium latum (Broad Fish Tapeworm) : stages   • Infective - ingestion of larvae in infected undercooked freshwater fish • Diagnostic- demonstrate characteristic egg or proglottids (often in chains of a few inches- feet) in feces (may NOT be seen in flotation techniques; operculated ova may sink)  
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Diphyllobothrium latum (Broad Fish Tapeworm) : clinical disease   • Can cause megaloblastic anemia, since vitamin B12 is absorbed by worm • Sparganosis- disease caused by drinking H2O containing infected copepod. Larva develops in human who is the intermediate host instead of the fish  
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Echinococcus granulosus (Hydatid Tapeworm): adult   • Very small; 3-6cm long • Consists of only 3 proglottids- immature, mature, and gravid • Scolex has an armed rostellum  
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Echinococcus granulosus (Hydatid Tapeworm): normal life sycle   • Normal life cycle • Sheep (intermediate host) ingest eggs • Dog (definitive host) infected from eating infected viscera of butchered animals  
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Echinococcus granulosus (Hydatid Tapeworm): infective stage   • Infective stage- man (intermediate host) ingests egg  
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Echinococcus granulosus (Hydatid Tapeworm): diagnostic stage part 1   • Hydatid cysts seen in routine x-rays or exploratory surgery (form in various parts of the body- most commonly the liver, lungs, brain, heart). Hydatid cysts contains hydatid sand  
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Echinococcus granulosus (Hydatid Tapeworm): diagnostic stage part 2   •• Serological test (ELISA, IHA) • Casoni’s skin test- intradermal test using hydatid cyst fluid as antigen  
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Dipylidium caninum (Dog Tapeworm)   • Found worldwide in children  
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Dipylidium caninum (Dog Tapeworm): adult   • Scolex has an armed rostellum with many rows of thorn-shaped hooks • Proglottids resemble pumpkin/cucumber seeds when moist and rice grain when dry  
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Dipylidium caninum (Dog Tapeworm): normal life cycle   • Dog and cat ingest infected fleas containing larvae  
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Dipylidium caninum (Dog Tapeworm): stages   • Infected stage- human (accidental intermediate host) ingests infected flea • Diagnostic stage- demonstrate characteristic egg packet (5-10 eggs are in a hylaline non-cellular egg sac) in feces  
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Trematodes (Flukes): general info   • Flat leaf-shaped organisms • Hermaphroditic- contain both male and female reproductive parts (except Schistosomes) • Required an intermediate host • Snail is always 1st intermediate host • Eggs are operculated (except Schistosomes)  
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Fasciolopsis buski (Giant Intestinal Fluke)   • Found in Asia  
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Fasciolopsis buski (Giant Intestinal Fluke): adults   • Large fluke, 75mm long by 20mm wide • Poorly developed suckers • Live in intestines of pigs and humans  
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Fasciolopsis buski (Giant Intestinal Fluke): stages   • Infective stage- ingestion of raw aquatic vegetation (Ex. Water chestnuts) with encysted metacercariae • Diagnostic stage- demonstration of characteristic eggs in feces (eggs resemble F. hepatica)  
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Fasciolopsis buski (Giant Intestinal Fluke): clinical disease   • Diarrhea, epigastric pain • Symptoms relate to number of worms present  
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Fasciola hepatica (Liver Fluke)   • Sheep and cattle are reservoir host; therefore, high incidence in sheep raising countries  
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Fasciola hepatica (Liver Fluke): adults   • Large fleshy fluke 30mm long 13mm wide • Live in bile ducts of liver  
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Fasciola hepatica (Liver Fluke): stages   • Infective stage- ingestion of raw aquatic vegetation (Ex. Water chestnuts) with encysted metacercariae • Diagnostic stage- detect characteristic eggs in feces (eggs resemble F. buski)  
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Fasciola hepatica (Liver Fluke): clinical disease   • Larvae elicit inflammatory response in liver • Stone formation may occur • Eosinophilia may occur during larvae migration  
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Clonorchis sinensis (Chinese Liver Fluke)   • Found in Japan, Korea, China, Taiwan, and Vietnam (cats and dogs serve as reservoir hosts)  
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Clonorchis sinensis (Chinese Liver Fluke): adults   • Long and narrow, 20mm long and 3-5mm wide • Live in bile ducts of liver  
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Clonorchis sinensis (Chinese Liver Fluke): stages   • Infective stage- ingestion of raw fish infected with metacercariae • Diagnostic stage- demonstration of characteristic eggs in feces (operculated with shoulders and small knobs at abopercular end)  
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Paragonimus westermani (Oriental Lung Fluke)    
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