1. Quantitative Flotation (McMaster)

2. 4 gram 56 ml StirWait 30 min.  10 ml Centrifuge 1200 g 5-7 min Gauze 30x30 cm Remove supernatant with pipette Add 3ml flotation liquid and mix with pipette Fill a reading chamber Wait 5 min. Read 3 vertical lines at x200 magnification and note species and Count numbers Multiplication factor: 47 3 ml

3. x20 Added flotation liquid 1,5 / 3 / 6 ml EPG (Eggs Per Gram) = Count X Multiplication factor example. EPG = 2 x 83= 166 Visual area ObjectiveVolume pellet is diluted in (ml) 1 visual area 3 visual areas 1 column 3 columns Total chamber x41,5 377126561910 3,0 306102501710 6,0 25384461510 x101,5 23587861394610 3,0 19106371254210 6,0 15785261143810 x201,5 943131442789310 3,0 763925472508310 6,0 631621052277610 x401,5 377261257555618510 3,0 305581018650016710 6,0 25254841845515210 x1001,5 23578578595138946310 3,0 19098663662125041710 6,0 15784052613113637910 3 ml

4. 40 U = 8

5. Sedimentation (heavy parasite eggs)

6. Fasciola hepatica Dicrocoelium dendriticum Fasciola buski Opisthrchis noverca 3 g faeces + 50 ml water gauze Fill with water  Wait 10 minRemove supernatant Wait 10 minResuspend in water Remove supernatant Add 1 drop of dye + mix Add drop to slide + cover glass + read Nematodirus spp. Sedimentation („Trematode“ eggs) Method for heavier eggs that do not flotate in common flotation fluids

7. Baermann (lungworms & L3)

8. 5-10 g faeces Fill with water Centrifuge 1000 rpm, 2 min Remove excess water Baermann (lungworms & L3 larvae) Harvesting of larvae from faecal sample water Wait 24 hours (no direct sun) Fold gauze and put in syringe Remove faeces and excess water Transfer to 14 ml centrifuge tube Transfer to petri dish and examine under microscope

9. Baermann (lungworms) Harvesting of larvae from faecal sample Dictyocaulus viviparus (390-450 µm) Dictyocaulus filaria (550-585 µm) Larvae often curved Sluggish movement Muellerius capillaris (300-320 µm) Posterior end of body often curled Protostrongylus (320-400 µm) Posterior half of body may be curled Cystocaulus (340-480 µm) Additional dorsal and ventral spines on middle part of tail Sheep Cattle

10. Baermann (L3 larvae) Harvesting of larvae from faecal sample Trichostrongylus (560-796 µm) 16 gut cells, Head tapered, Sheath forms short cone http://www.rvc.ac.uk/review/parasitology/RuminantL3/Introduction.htm Cooperia (666-956 µm) 16 gut cells, Head square with refractile bodies, Sheath tail medium tapering (Cooperia oncophora), or finely pointed and refractile (Cooperia curticei type) Ovine Bovine Ostertagia Strongyloides (650-850 µm) Oesophagus extends nearly half length of body, Sheath absent, Larval tail notched Oesophagostomum (726-923 µm) / Chabertia (650-789 µm) Approx. 32 gut cells, Head broad and rounded, Sheath tail filamentous, Larval tail not notched or lobed

11. Ziehl-Neelsen [acid fast staining] (Cryptosporidium spp.)

12. Zeihl-Neelsen (Cryptosporidium spp.) Detection of C. spp. oocysts in faeces Add small amount of feaces to slide Add name with pencil Smear feaces using second slide Dry Fixate methanol (+2% HCl) 5 min Color carbolic fuxine 20 min Remove color 10% H2SO4 10 sec Wash Water 10 sec Repeat 2 times Color Brilliant Green 2.5% 5 min Wash Water 10 sec Read with x400 magnification 0 = no oocysts + = 1-5 oocysts ++ = 6-25 oocysts +++ = >25 oocysts

13. Ziehl-Neelsen (Cryptosporidium spp.) Detection of C. spp. oocysts in faeces Read with x400 magnification 0 = no oocysts + = 1-5 oocysts ++ = 6-25 oocysts +++ = >25 oocysts SpeciesSize (µm)Hosts Cryptosporidium parvum5.0 x 4.5Mammals, primarily cattle and humans Cryptosporidium hominis5.0 x 4.5Humans, monkeys, also other animals Cryptosporidium andersoni7.4 x 5.5Cattle and other ruminants Cryptosporidium canis5.0 x 4.7Dogs, also other animals Cryptosporidium felis4.6 x 4.0Cats, also other animals Cryptosporidium suis4.6 x 4.2Pigs, also found in humans Cryptosporidium baileyi6.3 x 5.0Birds Cryptosporidium meleagridis4.5 x 4.0Birds Cryptosporidium serpentis6.2 x 5.3Reptiles http://courses.cit.cornell.edu/biomi290/microscopycases/crypto/index.htm