Mite preparations for identifications Day – 3 Felicity Crotty 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 Soil Mesofauna = Springtails and Mites Mites (Acari) • • • • Three main orders Mesostigmata Oribatids Prostigmata • 45,000 described spp • Approx 64 spp in 1 grassland Stable Community Composition: Mites Diversity mites in grassland (+/- 0.05 s.e.) Diversity of mites in willow woodland (+/- 0.05 s.e.) 9% 11% 16% 31% 23% 14% 61% 35% ASTIGMATA MESOSTIGMATA ASTIGMATA MESOSTIGMATA ORIBATID PROSTIGMATA ORIBATID PROSTIGMATA 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 stored in alcohol Mite soaked overnight in 90% lactic acid (on warmer) Mite placed on slide in pva, coverslip edges sealed with dpx “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 stored in alcohol Mite soaked overnight in 90% lactic acid (on warmer) Mite placed on slide in PVA, coverslip edges sealed with DPX Identification • What level? - Order (Collembola or Mite) - Lineage (Mesostigmata/Oribatida) - Supercohort (Macropyline/Brachypyline) - Cohort (Palaeosomata/Mixonomata) - Superfamily (Phthiracaroidea/Lohmannioidea) Sub-Class Acari Super-Order Order Parasitiformes Holothrida Opilloacarida Trigynaspida Acariformes Mesostigmata Sub-Order Sphaerolichida Endeostigmata Monogynaspida Palaeosomata Eupodides Parhyposomata Labidostommatides Enarthronota Cohort Microgynina Eleutherengonides Brachypylina Astigmata Uropodina Heterostigmata Antennophorina Sub-Cohort / Infra-Order Arctacariae Parasitengonina Gamasina Euptyctima Epicriiae Parasitiae Raphignathina Anystina Heterozerconina Heatherellina Anystides Desmonomata Mixonomata Cercomegistina Prostigmata Oribatida Sejida Super-Cohort Trombidiformes Sarcoptiformes Ixodida Dermanyssiae Dichosomata Pycnonticae Poronoticae Methods of identification • • • • Computer based key “Lucid” Available online “Major mite taxa” home page http://itp.lucidcentral.org/id/mites/invasive_mite/I nvasive_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 Different Lucid Key Greater scope - COHORT Mesostigmata, Oribatid and Prostigmata. http://keys.lucid central.org/keyserver/player.jsp ?keyId=42 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 Dichotomous keys • Greater depth Orders… Lineages… Families… Genus… Species Sub-Class Acari Super-Order Order Parasitiformes Holothrida Opilloacarida Trigynaspida Acariformes Mesostigmata Sub-Order Sphaerolichida Endeostigmata Monogynaspida Palaeosomata Eupodides Parhyposomata Labidostommatides Enarthronota Cohort Microgynina Eleutherengonides Brachypylina Astigmata Uropodina Heterostigmata Antennophorina Sub-Cohort / Infra-Order Arctacariae Parasitengonina Gamasina Euptyctima Epicriiae Parasitiae Raphignathina Anystina Heterozerconina Heatherellina Anystides Desmonomata Mixonomata Cercomegistina Prostigmata Oribatida Sejida Super-Cohort Trombidiformes Sarcoptiformes Ixodida Dermanyssiae Dichosomata Pycnonticae Poronoticae Sub-Class Acari Super-Order Order Parasitiformes Holothrida Opilloacarida Trigynaspida Acariformes Mesostigmata Sub-Order Sphaerolichida Endeostigmata Monogynaspida Palaeosomata Eupodides Parhyposomata Labidostommatides Enarthronota Cohort Microgynina Eleutherengonides Brachypylina Astigmata Uropodina Heterostigmata Antennophorina Sub-Cohort / Infra-Order Arctacariae Parasitengonina Gamasina Euptyctima Epicriiae Parasitiae Raphignathina Anystina Heterozerconina Heatherellina Anystides Desmonomata Mixonomata Cercomegistina Prostigmata Oribatida Sejida Super-Cohort Trombidiformes Sarcoptiformes Ixodida Dermanyssiae Dichosomata Pycnonticae Poronoticae Soil Mite or Tick?! Parasitiformes Mesostigmata Ixodida Soil Mite or Tick?! Parasitiformes Mesostigmata - 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 Ixodida - Greater than ~5mm - Peritremes and stigmatal opening absent - Subcapitulum with denticulate hypostome - Palps usually 4 or fewer segments, without apotele - Tritosternum absent Soil Mite or Tick?! Parasitiformes Mesostigmata Ixodida 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 (or Parasitiformes) (Figure) Chelicerae Chelate-dentate 6 free segments: Tarsus, tibia, genu, femur, trochanter, coxae Stigmata and peritreme Probably male as epigynal shield not distinct, or hinged 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 Figure 1 Oribatida Rutella Chelicerae Chelate Tracheal system Oxotaxic pores Genital and anal shields But this is also an Oribatid… AND this is an Oribatid too! Prostigmata Chelicerae and pedipalps modified Stigmata at base of gnathosoma Body weakly sclerotised 5 free segments: Tarsus, tibia, genu, femur, trochanter. CHARACTER PARASITIFORMES (MESOSTIGMATA) SARCOPTIFORMES (ORIBATIDS & ASTIGS) TROMBIDIFORMES (PROSTIGMATA) Chelicerae Chelate-dentate (except parasites) Usually chelate-dentate Often modified (spikes, stylets) Hypostomal groove Present Absent Absent Tritosternum Usually present Absent Absent Gnathosomal tectum Present Absent Absent Palps Usually well developed, limb-like 5-segmented in Oribatida; 2-segmented in Astigmata Often reduced or modified Palp apotele Present Absent Absent Palp thumbclaw Present Absent Sometimes present Dorso-sejugal suture Absent Present Often indistinct Eyes Absent (except Allothyridae) Absent (rare exceptions) Sometimes present Trichobothria Absent Present (except Nothroidea) Often present Dorsal & ventral plates Usually 1 or 2 dorsal plates and various ventral plates Usually well developed dorsal and ventral plates Often absent or small (numerous exceptions) Colour Usually brown to red-brown Usually brownish to black Often white, yellow, green, red Genital & anal openings Genital plate undivided, anal plate divided With divided plates Usually without plates Genital papillae Absent Present Often present Lateral peritreme Usually present Absent Absent Lateral opisthosomal gland Absent Usually present Absent Coxae free Fused to ventral body Fused to body CHARACTER PARASITIFORMES (MESOSTIGMATA) SARCOPTIFORMES (ORIBATIDS & ASTIGS) TROMBIDIFORMES (PROSTIGMATA) Chelicerae Chelate-dentate Usually chelate-dentate Often modified (spikes, stylets) Hypostomal groove Present Absent Absent Tritosternum Usually present Absent Absent Gnathosomal tectum Present Absent Absent Palps Usually well developed, limb-like 5-segmented in Oribatida; 2-segmented in Astigmata Often reduced or modified Palp apotele Present Absent Absent Palp thumbclaw Present Absent Sometimes present Dorso-sejugal suture Absent Present Often indistinct Eyes Absent (except Allothyridae) Absent (rare exceptions) Sometimes present Trichobothria Absent Present (except Nothroidea) Often present Dorsal & ventral plates Usually 1 or 2 dorsal plates and various ventral plates Usually well developed dorsal and ventral plates Often absent or small (numerous exceptions) Colour Usually brown to red-brown Usually brownish to black Often white, yellow, green, red Genital & anal openings Genital plate undivided, anal plate divided With divided plates Usually without plates Genital papillae Absent Present Often present Lateral peritreme Usually present Absent Absent Lateral opisthosomal gland Absent Usually present Absent Coxae Free Fused to ventral body Fused to body CHARACTER PARASITIFORMES (MESOSTIGMATA) SARCOPTIFORMES (ORIBATIDS & ASTIGS) TROMBIDIFORMES (PROSTIGMATA) Chelicerae Chelate-dentate Usually chelate-dentate Often modified (spikes, stylets) Hypostomal groove Present Absent Absent Tritosternum Usually present Absent Absent Gnathosomal tectum Present Absent Absent Palps Usually well developed, limb-like 5-segmented in Oribatida; 2-segmented in Astigmata Often reduced or modified Palp apotele Present Absent Absent Palp thumbclaw Present Absent Sometimes present Dorso-sejugal suture Absent Present Often indistinct Eyes Absent (except Allothyridae) Absent (rare exceptions) Sometimes present Trichobothria Absent Present Often present Dorsal & ventral plates Usually 1 or 2 dorsal plates and various ventral plates Usually well developed dorsal and ventral plates Often absent or small (numerous exceptions) Colour Usually brown to red-brown Usually brownish to black Often white, yellow, green, red Genital & anal openings Genital plate undivided, anal plate divided With divided plates Usually without plates Genital papillae Absent Present Often present Lateral peritreme Usually present Absent Absent Lateral opisthosomal gland Absent Usually present Absent Coxae free Fused to ventral body Fused to body CHARACTER PARASITIFORMES (MESOSTIGMATA) SARCOPTIFORMES (ORIBATIDS & ASTIGS) TROMBIDIFORMES (PROSTIGMATA) Chelicerae Chelate-dentate Usually chelate-dentate Often modified Hypostomal groove Present Absent Absent Tritosternum Usually present Absent Absent Gnathosomal tectum Present Absent Absent Palps Usually well developed, limb-like 5-segmented in Oribatida; 2-segmented in Astigmata Often reduced or modified Palp apotele Present Absent Absent Palp thumbclaw Present Absent Sometimes present Dorso-sejugal suture Absent Present Often indistinct Eyes Absent (except Allothyridae) Absent (rare exceptions) Sometimes present 1 to 2 pairs) Trichobothria Absent Present (except Nothroidea) Often present Dorsal & ventral plates Usually 1 or 2 dorsal plates and various ventral plates Usually well developed dorsal and ventral plates Often absent or small (numerous exceptions) Colour Usually brown to red-brown Usually brownish to black Often white, yellow, green, red Genital & anal openings Genital plate undivided, anal plate divided With divided plates Usually without plates Genital papillae Absent Present Often present Lateral peritreme Usually present Absent Absent Opisthosomal gland Absent Usually present Absent Coxae free Fused to ventral body Fused to body 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 Soil Acarology 3 week course Summer 2014 http://www.biosci.ohiostate.edu/~acarolog/summerProgram/index.html Acknowledgments Soil Mite Experts Cal Welbourne Valerie Behan-Pelletier Roy Norton David Walter