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Parasitology #1

Classification of Internal Parasites of Veterinary Interest Nematodes, Trematodes, Cestodes, and Protozoa
What's an example of a nematode? Roundworm
Describe a nematode Cylindrical, non-segmented helminths ; a complete digestive tract with oral and anal openings, and taper at both ends
What's an example of a Trematode? Fluke
Describe a Trematode. Leaf shaped with an outer tegument which may be smooth or spiny, Possess an anterior oral sucker and a posterior ventral sucker. Hermaphrodites, life cycle involves one or more hosts (indirect life cycle), and a developed digestive system
Where do Trematodes live? – Digestive tract – Liver – Bile duct – Gall bladder – Lungs – Bladder and ureter
What's an example of a cestode? Tapeworms
What do tapeworms look like? flat, segmented, ribbon-like bodies and lack an alimentary canal
Where do adult tapeworms live? The digestive tract
What are examples of Protozoa? ciliates, coccidia, and flagellates
Define Protozoa Unicellular organisms that are classified on the basis of locomotion
Where do protozoa live? the epithelial cells of the intestinal tract
Classification of External Parasites of Veterinary Interest insects and arachnids
What are examples of insects? flies, fleas and lice
What are examples of arachnids? ticks, mites and spiders
What methods are used to test for internal parasites? Testing blood, urine, fecal, and vaginal/cervical mucus samples
Fecal tests Gross Examination of Feces Fecal Flotation Procedure Centrifugal Fecal Flotation Procedure Fecal Sedimentation Procedure Direct Fecal Smear Procedure Examination of Tapeworm Segments
What methods are used to collect fecal samples? voided samples direct samples pooled samples
voided samples animal defecates and sample is collected
direct samples a fecal loo or gloved finger, hand or arm
pooled samples randomly collected samples from an area in which several animals are housed
handling and storage of feces use fresh samples, store in clean, labeled container (owner's name, date, pet's name) and keep refrigerated
Why do we refrigerate fecal samples? parasites break down quickly, some hatch and are easily confused with other parasites, and larvae break down quickly too
What precautions should be taken during fecal exams? use PPE, wash hands frequently, no food or drink in the lab, clean in between exams, handle with care (possible zoonotic diseases)
Gross Examination of Feces Consistency color blood mucus adult parasites or tapeworm segments
What materials are needed for a Fecal float? flotation solution ( sodium nitrate or zinc sulfate), flotation device, applicator stick, microscope slide and cover, mixing cup and filter (if using centrifuge tube)
What is the procedure for a Fecal Float? 1. mix 2 grams of feces and flotation solution in fecalizer until no large pieces remain 2. filter emulsion through fecalizer 3. fill vial or fecalizer to form meniscus 4. place coverslip over top 5. allow fecalizer to sit 10-20 min
What is the procedure for a Fecal Float? (cont.) 6. remove coverslip and place on microscope slide 7. examine under microscope
What is the process for a Centrifugal Fecal Flotation Procedure? 1. Prepare the fecal emulsion as described for simple flotation. 2. Strain the emulsion through the cheesecloth into the centrifuge tube and fill the tube nearly to the top. Balance the centrifuge. Mark tubes for identification.
What is the process for a Centrifugal Fecal Flotation Procedure? (cont.) 3. Centrifuge tube for 5 minutes at 1500 RPM. 4. Gently remove tube from centrifuge and place in rack. Touch a wire loop or a glass rod to the surface of the liquid. Transfer drop to a microscope slide. Apply coverslip and examine under the microscope.
What materials are needed for a Fecal Sedimentation Procedure? Beaker or Wax-lined paper cup Gauze Sponge Tongue Depressors Centrifuge and 15 ml Centrifuge Tubes Pipette Microscope Slide and Coverslip
What is the process for a Fecal Sedimentation Procedure? 1. Using a tongue depressor, mix ½ tsp of feces with tap water or saline in a cup or beaker. Strain the mixture through the gauze into the centrifuge tube.
What is the process for a Fecal Sedimentation Procedure? (cont.) 2. Balance the centrifuge and spin at 1500 RPM. If centrifuge is unavailable, allow mixture to sit undisturbed for 20-30 minutes. 3. Pour off the supernatant without disturbing the sediment at the bottom.
What is the process for a Fecal Sedimentation Procedure? (cont.) 4. Using the pipette, transfer a small amount of the top layer of sediment to a microscope slide. If the drop is too thick, dilute it with a drop of water. 5. Examine under the microscope at 10X and 40X.
What materials are needed for Direct Fecal Smear Procedure? Microscope Slide and Coverslip Applicator Stick
What is the process for a direct Fecal smear procedure? 1. Place a drop of saline or water in the middle of a slide and an equal amount of feces next to it. 2. Thoroughly mix the feces and water with the applicator stick to form an emulsion of the two substances.
What is the process for a direct Fecal smear procedure? (cont.) 3. Make a smear on the surface of the slide. Be sure that your smear is not too thick. 4. Remove any large fecal particles with the stick and place a coverslip over the smear. 5. Examine the smear under a microscope.
Examination of Tapeworm Segments Dried segments must be soaked in saline for 1 to 4 hours. Once segments are rehydrated, the tapeworm may be identified by: • Size • Shape and • Eggs contained within
How do you identify tapeworm species if there are no eggs in the segment? Some segments may not contain eggs. In order to identify the tapeworm species, the segment is crushed between two glass slides and examined under a microscope. The species may be identified by its calcareous bodies
What methods are used to examine blood? Direct Microscopic Examination of Whole Blood, Thin Blood smear, Filter Technique for Microfilaria, Buffy Coat Method
What materials are needed for Direct Microscopic Examination of Whole Blood? Blood (preferably fresh blood collected in the afternoon as D. immitis are more numerous in the circulating blood at this time), Microscope Slide, Coverslip
What is the process of Direct Microscopic Examination of Whole Blood? a single drop from the needle onto the center of a slide and cover it with a coverslip. 2. Examine under the microscope with the 10X objective. Watch for localized areas of movement among blood cells, which may indicate the presence of parasites.
What materials are needed for a thin blood smear? Blood, Microscope Slides, Blood Smear Stain (Diff-Quik, Wright’s, Giemsa)
What is the process for a Thin Blood Smear? 1. Add a drop of blood near one end of the slide. 2. Place the end of a second slide near the middle of the first slide at a 45-degree angle. gently pull the slide back into the drop of blood.
What is the process for a Thin Blood Smear? (cont.) 3. Holding the first slide firmly, smoothly push the second slide across and off the end of the first slide. This should form a thin smear on the first slide, with a nice feathered edge.
What is the process for a Thin Blood Smear? (cont.) 4. Allow the slide to air dry, then stain. After staining, the slide can be examined using the oil-immersion objective.
What materials are needed for Filter Technique for Microfilaria? Filter Device Filter Paper 9 ml of Lysing Solution 1 ml of Whole Blood
What is the process for Filter Technique for Microfilaria? 1.Mix 1 ml of blood with 9 ml of lysing solution in a syringe. 2. Assemble the filter cartridge. 3. Inject the blood and lysing solution through the filter. 4. follow up with 10 ml of tap water to flush
What is the process for Filter Technique for Microfilaria? (cont.) 5. The filter is then gently transferred to a microscope slide and stained with 2 drops of stain. 6. Cover with a coverslip and examine under 10X for microfilaria. They will look like small blue threads – narrow at each end
What materials are needed for the buffy coat method? hematocrit tube and sealant hematocrit centrifuge
What is the process for Buffy Coat method? 1. Fill the hematocrit tube with the blood sample and seal one end. 2. Centrifuge for 5 minutes. 3. Place the hematocrit tube under a microscope and examine directly above the buffy coat under 10X and 40X looking for microfilaria movement.
How do you diagnose for Ectoparasites? skin scraping, cellophane tape prep, swabs
What materials are needed for skin scraping? • Electric Clipper with #40 Blade • #10 Scalpel Blade • Mineral Oil in Small Dropper Bottle • Microscope Slide with a Drop of Mineral Oil
What is the process for skin scraping? 1. Shave site to be scraped. 2. Dip scalpel blade in a drop of mineral oil on the slide. 3. Hold blade between the thumb and forefinger, keep blade perpendicular to the lesion to avoid accidental incision.
What is the process for skin scraping? (cont.) 4. Scrape the skin until a small amount of capillary blood oozes from the scraped area. 5. Spread scraped debris into the drop of mineral oil on the slide and apply a coverslip. 6. Examine under 4X and 10X objectives.
What materials are needed for Cellophane Tape Preparation? • Transparent Adhesive Tape • Wooden Tongue Depressor • Microscope Slide • Mineral Oil
What is the process for Cellophane Tape Preparation? 1. Place the tape in a loop around one end of the tongue depressor with the adhesive side out. 2. Press the tape on the tongue depressor firmly against the skin to pick up epidermal debris.
What is the process for Cellophane Tape Preparation? (cont.) 3. Place a ribbon of mineral oil along the microscope slide then apply the adhesive side of the tape to the mineral oil. 4. Examine slide under a microscope.
Created by: kcrabtree507



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