Flotation Method

Medical Parasitology Lab. Flotation MethodConcentration techniquesRaed Z. Ahmed, Medical Parasitology Lab.,2012-2013Prepared By:Mr. Raed Z. Ahmed1Concentration MethodsSedimentation methodModified Formal- Ether sedimentation techniqueAcid- Ether sedimentation techniqueFlotation methodSaturated Salt Solution techniqueSheathers Sugar Centrifugal Flotation techniqueZinc Sulphate Centrifugal Flotation techniqueRaed Z. Ahmed, Medical Parasitology Lab.,2012-2013Flotation techniqueThese method use the high specific gravity of a solution to float the lighter ova and cyst. They can be improved by centrifugation. Advantage:Easy to perform .Disadvantage:Delay in examination can result in distortion.Larvae and some fluke eggs do not concentrate.Frequent checking of specific gravity.Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Saturated salt solutionFlotation MethodsRaed Z. Ahmed, Medical Parasitology Lab.,2012-2013Materials and Method Boil granular sodium chloride in excess in water to produce a saturated solution which when cooled has a specific gravity of 1.18 - 1.2.Half fill a wide- mounted flat bottomed container with the saturated salt solution.Emulsify 1gm of feces in the solution and strain it to remove the debris from the surface.Pour the filtrate into meniscus and fill it to the top with saturated salt solution.Lay a glass slide or large coverslip over the top, avoiding any bubbles being trapped.Leave for 20 min before quickly inverting the slide.Scan for ova using the 10x objectives.Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Saturated Salt solution techniqueAdvantages:It is cheap preparation using simple apparatus.It concentrates nematode ova well.Disadvantages:It doesnt concentrate cysts.Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Sheathers sugar centrifugal flotation techniqueFlotation MethodsRaed Z. Ahmed, Medical Parasitology Lab.,2012-2013Materials and MethodSheathers sugar solution:Table sugar -------------------------------------- 500gmDistilled water ---------------------------------- 320mlPhenol crystal ( melt in hot water bath) ----- 6.5gm

Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Procedures Soften 1gm of feces with water to a soft.Strain the aqueous suspension through a wire sieve.Mix 1 part aqueous suspension with 2 part of Sheather's sugar solution.Pour into a centrifuge tube, centrifugation 1500 rpm for 10 minutes.Pour the supernatant into a meniscus and add a sufficient solution to bring the meniscus to the top.Place a coverslip and wait for 10 minutes.Examine under microscope.Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Advantages:Reveals most nematode eggs and protozoan cyst.

Disadvantages:Flukes eggs and tape worm eggs are not demonstrate well.Also most nematode larvae are not demonstrate well.Sheathers Sugar solution techniqueRaed Z. Ahmed, Medical Parasitology Lab.,2012-2013Balantidium coliIntestinal Protozoa Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Balantidium coliIs a parasitic species of ciliate protozoa that cause balantidiasis or Balantidium dysentery. Balantidium coli has two developmental stages, a trophozoite stage and a cyst stage.The cyst is the infective stage measures about 50 to 70 m in diameter, characterized by the presence of a large kidney-shaped macronucleus.The trophozoite is oval with 2 nuclei and measures approximately 50 to 100 m long and 40 to 70 m wide.

Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013B.coli Cyst

Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013B. coli Trophozoite

Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Cryptosporidium parvumIntestinal Protozoa Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Cryptosporidium parvumInfect human and most mammals.The infective stage is oocyst containing sporozoites measuring 5u in diameter.The diagnostics stage is sporocyst containing 4 sporozoites.Diagnosis:Detecting oocyst in stool.Acid-fast stain. Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Cryptosporidium parvum oocyst

Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Toxoplasma gondiiTissue ProtozoaRaed Z. Ahmed, Medical Parasitology Lab.,2012-2013Toxoplasma gondiiHabitat: reticulo-endothelial system, monocyte, and muscle fiber and cause toxoplasmosis.Humans can acquire Toxoplasma gondii infection by ingestion cyst or sporylated oocystCyst: precipitated in flesh of cow.Sporulated oocyst: cat feces.Intermediate host: Human.Definitive host: Cat.The tachyzoites are crescent-shaped and measure about 5 m in length.Diagnostic stages:Diagnosis also can be done by detection antibody IgG, IgM, IgE and IgA.

Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013

Tachyzoites

Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Trichomonas vaginalisUrogenital Protozoa Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Trichomonas vaginalisIs the most common cause of vaginitis, inhabit the urogenital systemThere is no cyst stage but only have trophozoite stage.The trophozoite measures about 15 x 10 m. The trophozoite has a single nucleus and four flagella and undulating membrane. Diagnosis:Male: finding trophozoite in urethral prostatic dischargeFemale: finding trophozoite in vaginal discharge. Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013T. vaginalis Trophozoite

Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Hymenolepis nanaIntestinal Cestodes Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Hymenolepis nanaThis is one of the most common cestodes of humans, especially children. And its the smallest tape worm in man, also called dwarf tape worm.Eggs measures 30-37u, and have double membrane and contain six hooked oncosphere.The scolex of H. nana has four suckers and a short rostellum with hooks.Infective stage: Eggs.Diagnosis: Stool examination to detect the eggs.Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013Hymenolepsis diminuta is primarily a tapeworm of rodents. Humans are infected following the accidental ingestion of arthropods containing the cysticercoid stage of the parasite. The ova (60 to 70 m in diameter) are passed in the stool of infected persons. The ova contain a six-hooked oncosphere and no polar filaments.The scolex of H. diminuta has four suckers and a rostellum without hooks25H. nana egg

Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013H. nana adult worm

Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013H. nana scolex

Raed Z. Ahmed, Medical Parasitology Lab.,2012-2013