Quantitative Flotation (McMaster)

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

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, , , x101, , , x201, , , x401, , , x1001, , , ml

40 U = 8

Sedimentation (heavy parasite eggs)

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

Baermann (lungworms & L3)

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

Baermann (lungworms) Harvesting of larvae from faecal sample Dictyocaulus viviparus ( µm) Dictyocaulus filaria ( µm) Larvae often curved Sluggish movement Muellerius capillaris ( µm) Posterior end of body often curled Protostrongylus ( µm) Posterior half of body may be curled Cystocaulus ( µm) Additional dorsal and ventral spines on middle part of tail Sheep Cattle

Baermann (L3 larvae) Harvesting of larvae from faecal sample Trichostrongylus ( µm) 16 gut cells, Head tapered, Sheath forms short cone Cooperia ( 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 ( µm) Oesophagus extends nearly half length of body, Sheath absent, Larval tail notched Oesophagostomum ( µm) / Chabertia ( µm) Approx. 32 gut cells, Head broad and rounded, Sheath tail filamentous, Larval tail not notched or lobed

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

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

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