1. Mite preparations for identifications Day – 3 Felicity Crotty

2. Collection Most common method for mite collection is the use of Tullgren funnels. Although others possible – floatation / pooters. Collection / storage in 70% alcohol best method (unless want to further experiment) Saturated salt solution also used

3. Soil Mesofauna = Springtails and Mites Mites (Acari) • 45,000 described spp • Approx 64 spp in 1 grassland • Three main orders • Mesostigmata • Oribatids • Prostigmata

4. Stable Community Composition: Mites

5. Mite stored in alcohol Preservation Dissecting microscope to sort through fauna Compound microscope to observe external structures for key Highly sclerotised mites need to be “cleared” and disected before mounting on slides This is to make “permanent” slides Mite soaked overnight in 90% lactic acid (on warmer) Mite placed on slide in pva, coverslip edges sealed with dpx

6. Mite stored in alcohol “Quick” Preservation Mites were left in eppendorfs on a plate heater (lowest setting) overnight PVA = polyvinyl alcohol (I used to get mine from Sigma) I used 1 drop ~ 3 µl DPX = DPX Mountant (again from Sigma) Nail polish could also be used instead of DPX, but this does not last as long Mite soaked overnight in 90% lactic acid (on warmer) Mite placed on slide in PVA, coverslip edges sealed with DPX

7. Identification What level? - Order (Collembola or Mite) - Lineage (Mesostigmata/Oribatida) - Supercohort (Macropyline/Brachypyline) - Cohort (Palaeosomata/Mixonomata) - Superfamily (Phthiracaroidea/Lohmannioidea)

8. Sub-Class Acari Super-Order Parasitiformes Acariformes Trombidiformes Holothrida Mesostigmata Sarcoptiformes Ixodida Opilloacarida Order Sphaerolichida Endeostigmata Monogynaspida Trigynaspida Prostigmata Oribatida Sub-Order Sejida Palaeosomata Eupodides Parhyposomata Labidostommatides Enarthronota Anystides Super-Cohort Eleutherengonides Brachypylina Desmonomata Mixonomata Astigmata Cercomegistina Microgynina Raphignathina Uropodina Anystina Cohort Heterozerconina Heterostigmata Antennophorina Heatherellina Parasitengonina Gamasina Euptyctima Epicriiae Pycnonticae Sub-Cohort / Infra-Order Arctacariae Dichosomata Poronoticae Dermanyssiae Parasitiae

9. Methods of identification • Computer based key “Lucid” • Available online • “Major mite taxa” home page • http://itp.lucidcentral.org/id/mites/invasive_mite/Invasive_Mite_Identification/key/Major_Mite_taxa/Media/Html/Home_Major_Mite_Taxa.html • Unfortunately doesn’t cover that much – Order: Opilioacarida, Holothyrida, and Ixodida; Suborder: Mesostigmata; Supercohort: Oribatida and Prostigmata. SIMILAR TO ISPOT KEYS – but computer needs Java

14. Different Lucid Key Greater scope • COHORT Mesostigmata, Oribatid and Prostigmata. http://keys.lucidcentral.org/key-server/player.jsp?keyId=42

15. Dichotomous Keys • Used Tiling Key & Hopkins • Paired statements of either words or images • Have to follow specific order • If character unknown / can’t see it easy to make a mistake and misidentify

16. Dichotomous keys • Greater depth • Orders… • Lineages… • Families… • Genus… • Species

18. Sub-Class Acari Super-Order Parasitiformes Acariformes Trombidiformes Holothrida Mesostigmata Sarcoptiformes Ixodida Opilloacarida Order Sphaerolichida Endeostigmata Monogynaspida Trigynaspida Prostigmata Oribatida Sub-Order Sejida Palaeosomata Eupodides Parhyposomata Labidostommatides Enarthronota Anystides Super-Cohort Eleutherengonides Brachypylina Desmonomata Mixonomata Astigmata Cercomegistina Microgynina Raphignathina Uropodina Anystina Cohort Heterozerconina Heterostigmata Antennophorina Heatherellina Parasitengonina Gamasina Euptyctima Epicriiae Pycnonticae Sub-Cohort / Infra-Order Arctacariae Dichosomata Poronoticae Dermanyssiae Parasitiae

19. Sub-Class Acari Super-Order Parasitiformes Acariformes Trombidiformes Holothrida Mesostigmata Sarcoptiformes Ixodida Opilloacarida Order Sphaerolichida Endeostigmata Monogynaspida Trigynaspida Prostigmata Oribatida Sub-Order Sejida Palaeosomata Eupodides Parhyposomata Labidostommatides Enarthronota Anystides Super-Cohort Eleutherengonides Brachypylina Desmonomata Mixonomata Astigmata Cercomegistina Microgynina Raphignathina Uropodina Anystina Cohort Heterozerconina Heterostigmata Antennophorina Heatherellina Parasitengonina Gamasina Euptyctima Epicriiae Pycnonticae Sub-Cohort / Infra-Order Arctacariae Dichosomata Poronoticae Dermanyssiae Parasitiae

20. Soil Mite or Tick?! Parasitiformes Mesostigmata Ixodida

21. Soil Mite or Tick?! Parasitiformes Mesostigmata Ixodida • Smaller than ~ 5mm • Peritremes and stigmatal opening present (at level of coxae II-IV) • Four pairs of setae on subcapitulum • Apotele with 2 or 3 tines • Tritosternum present • Female covered by 1-4 epigynal shields • Greater than ~5mm • Peritremes and stigmatal opening absent • Subcapitulum with denticulate hypostome • Palps usually 4 or fewer segments, without apotele • Tritosternum absent

22. Soil Mite or Tick?! Parasitiformes Mesostigmata Ixodida

23. Acari Key to main groups • Stigmata usually without noticeable peritremes, and not situated at each side of the body near coxae I-IV. FEMALES: Genital shield jointed to open at one lateral edge of shield …………………………………………………..(next) • Stigmata, one on each side of the body, situated in the region of coxae I-IV and usually provided with an elongate peritreme; chelicerae chelate-dentate mainly. FEMALES: Genital shield jointed to open at base. – Mesostigmata(orParasitiformes) (Figure)

24. Chelicerae Chelate-dentate 6 free segments: Tarsus, tibia, genu, femur, trochanter, coxae Stigmata and peritreme Probably male as epigynal shield not distinct, or hinged

25. Acari Key to main groups • Gnathosoma with conspicuous rutella; chelicerae chelate, rarely modified; pedipalps simple. Tracheal system opening through stigmata or ‘pores’ on various parts of the body or absent; Body weakly OR strongly sclerotized. – Oribatida • Tracheal system opening through a pair of stigmata situated on or near the base of the gnathosoma; chelicerae and pedipalps usually strongly modified. Body usually poorly sclerotized. – Prostigmata

26. Figure 1 Oribatida Rutella Chelicerae Chelate Tracheal system Oxotaxic pores Genital and anal shields

27. But this is also an Oribatid…

28. AND this is an Oribatid too!

29. Prostigmata Chelicerae and pedipalps modified Stigmata at base of gnathosoma 5 free segments: Tarsus, tibia, genu, femur, trochanter. Body weakly sclerotised

34. Best Keys? • Mesostigmatic mites of Britain and Ireland – Evans and Till (1979) Trans. Zool. Soc. Lond. 35, 139-270 • Oribatida – Norton and Behan-Pelletier (2009) In A manual of acarology. Krantz and Walter. • OR: Acarina: Oribatida – Norton (1990) In Soil Biology. Dindal • Acarina: Prostigmata – Kethley (1990) In Soil Biology. Dindal • Acarina: Astigmata – Philips (1990) In Soil Biology. Dindal

35. Soil Acarology 3 week courseSummer 2014 http://www.biosci.ohio-state.edu/~acarolog/summerProgram/index.html

36. Acknowledgments Soil Mite Experts Cal Welbourne Valerie Behan-Pelletier Roy Norton David Walter