New MacrochelePratums species (Acari, Mesostigmata, Macrochelidae) associated with burying beetles (Silphidae, Nicrophorus) in North America.

Burying beetles (Silphidae, Nicrophorus) are hosts to a broad diversity of mites (Acari), including several species of Macrocheles Latreille, 1829 (Mesostigmata, Macrochelidae). The macrochelid fauna associated with silphids primarily in North America was surveyed; in total, 1659 macrochelids representing seven species were collected from 112 Nicrophorus beetles representing nine host species. Three new species of Macrocheles were discovered during the survey and described as Macrocheles willowaesp. n., M. pratumsp. n., and M. kaijusp. n. The barcode region of cytochrome oxidase subunit I (COI) was amplified from the three new described species, as well as M. nataliae and M. praedafimetorum, and analysed in a small phylogeny.

Introduction

Carrion-feeding beetles () are associated with a diverse assemblage of mites, nematodes, and fungi. () species are large-bodied beetles, that breed and feed on decaying organic matter, most often vertebrate carcasses (Anderson and Peck 1985). There are at least 60 extant species of worldwide, 22 of PageBreakwhich occur in the New World (Sikes et al. 2008, Sikes et al. 2016). beetles are unique amongst i<span class="Disease">nsects because most al">species provide biparental care and they bury small vertebrate carcasses in subterranean crypts (see Anderson and Peck (1985) for a summary of their life cycle). beetles are associated with a broad diversity of mites that can occur at high prevalences and abundances, with at least 14 al">species of mites representing four families collected off 95% of beetles in a given population (Wilson and Knollenberg 1987). The symbiotic relatio<al">span class="Disease">nship between silphids and their associated mites are poorly understood; however, the relationship may be a blend of commensalism and mutualism, as some mite species actively prey on eggs of carrion-feeding flies that compete with (Wilson and Knollenberg 1987).

The () are a cosmopolitan family of predaceous mites with at least 480 described species from 20 genera, occurring in a wide variety of organic substrates where they feed on nematodes and other microinvertebrates (Krantz 1998, Lindquist et al. 2009, Emberson 2010). There are about 320 described species of Latreille, 1829 () worldwide (Emberson 2010), many of which are phoretic as adult females on i<span class="Disease">nsects, including nine al">species which are associated with silphids (, , , , , , , , ) (Halliday 2000, Mašán 2003, Niogret et al. 2007, Perotti and Braig 2009). associated with silphids attach with their chelicerae to beetles din class="Chemical">al">spersing to and from carcasses, and they generally feed on nematodes, i<al">span class="Disease">nsect eggs and larvae, and other invertebrates on carrion (Wilson and Knollenberg 1987, Schwarz et al. 1998). Macrochelids phoretic on burying beetles are often overlooked and unstudied, resulting in a scarcity of information about their life history and novel species that remain to be described. A recent survey of tortoise mites (, ) on beetles (Knee et al. 2012) also uncovered three new species of associated with burying beetles. Herein, I propose and describe sp. n., sp. n., and sp. n., include a small phylogeny based on the barcode region of COI, and describe the diversity, abundance and host range of species found on throughout this survey.

Methods

Biological collections

Silphids were collected by various researchers across eight countries and 21 provinces or states (see acknowledgments). In Canada, most silphids were collected as bycatch from xylophagous beetle trapping by W.K. Specimens from other countries were collected primarily in <al">span class="Disease">pitfall traps, and others were hand-collected. Beetle specimens preserved in ethanol were shipped to Carleton University, and upon receipt specimens were placed in 95% ethanol and stored at -20°C. Using a dissecting microscope, silphids were identified to species using keys from Anderson and Peck (1985). The presence, abundance, PageBreakand attachment location of mesostigmatic mites was recorded. All mesostigmatic mites were removed and placed in a 0.5 ml microfuge tube with 95% ethanol and stored at -20°C for later identification and/or molecular analysis. Mites were slide-mounted in polyvinyl alcohol medium (6371A, BioQuip Products, Rancho Dominguez, California, United States of America (USA)) and cured on a slide warmer at 40°C for 3–4 days. Slide-mounted specimens were examined using a compound microscope (Leica DM 2500) with differential interference contrast illumination (DIC), and identified to species using the primary literature. Initial drawings of mites were made with pencil on paper using a camera lucida. Illustrations were later merged in Adobe Photoshop CS5 and redrawn in Adobe Illustrator CS5 using an Intuos 3 Graphics Tablet from WACOM Co., Ltd. (Saitama, Japan). Leg chaetotaxy is based on the system proposed by Evans (1963) and Evans and Till (1965). Idiosomal chaetotaxy follows the system of Lindquist and Evans (1965) as applied to macrochelids by Hyatt and Emberson (1988). Notation for glandular openings and poroids (proprioreceptors or lyrifissures) follows the system developed by Athias-Henriot (1969, 1971, 1975) and Johnston and Moraza (1991), as reviewed by Kazemi et al. (2014). Measurements were made from at least eight female specimens, all measurements are in micrometres (µm), and lengths presented with mean followed by the range in parenthesis. Type specimens are deposited in the Canadian National Collection of Insects, Arachnids, and Nematodes (CNC), at Agriculture and Agri-Food Canada, Ottawa, Ontario, Canada, and the Smithsonian Institution National Museum of Natural History.

Molecular methods

Genomic DNA was extracted from whole specime<span class="Disease">ns for 24 hours using a DNeasy Tissue kit (Qiagen, Inc., Santa Clara, California, USA). Following extraction, mites were removed from the extraction buffer, vouchers were-slide mounted, and genomic DNA was purified following the DNeasy Tissue kit protocol. PCR amplificatio<al">span class="Disease">ns were performed in a total volume of 25 µl, with 14.7 µl ddH2O, 2.5 µl 10× ExTaq buffer, 0.65 µl 25 mM MgCl2, 1.0 µl of each 10 µM primer, 2.0 µl 10 mM dNTPs, 0.15 µl ExTaq DNA polymerase, and 3 µl genomic DNA template. Primer pairs LCO1490 + HCO2198 (Folmer et al. 1994) were used to amplify a 689 bp fragment of the 5’–end of COI. PCR amplification was performed on an Eppendorf ep Gradient S Mastercycler (Eppendorf AG, Hamburg, Germany), using the following protocol: initial denaturation cycle at 94 °C for 3 min, followed by 45 cycles of 94 °C for 45 s, primer annealing at 45 °C for 45 s, 72 °C for 1 min, and a final extension at 72 °C for 5 min. Amplified products and negative controls were visualized on 1% agarose electrophoresis gels, and purified using pre-cast E-Gel CloneWell 0.8% SYBR Safe agarose gels (Invitrogen, Carlsbad, California, USA). Sequencing reactions followed the protocol of Knee et al. (2012), and sequencing was performed at the Agriculture and Agri-Food Canada, Eastern Cereal and Oilseed Research Centre Core Sequencing Facility (Ottawa, Ontario, Canada).

Sequence chromatograms were edited and contiguous sequences were assembled using Sequencher v5.3 (Gene Codes Corp., Ann Arbor, Michigan, USA). COI sequences were aligned manually in Mesquite v3.10 (Maddison and Maddison, 2016) according to the tra<span class="Disease">nslated amino acid sequence. COI sequences from (MBIOE1677-13, MBIOE1699-13) generated by the Barcode of Life Data Systems (BOLD) were included in the phylogeny. COI sequences on GenBank from two () al">species, and (FM179677, KR103486), were used as outgroup sequences. Sequences generated during this study have been submitted to GenBank (Table 1).

Table 1.

Collection information, host species records and mite abundance of () mites collected from () beetles, with GenBank accession numbers for COI.

Beetle number	Beetle species	Collection location	Coordinates	Collection date	Macrocheles species	Mite abundance	GenBank number
N002	Nicro. defodiens	CAN, ON, Algonquin P.P. 2	45.895, -78.071	16.vi.08	Macro. willowae sp. n.	1	–
N003	Nicro. defodiens	CAN, ON, Algonquin P.P. 2	45.895, -78.071	16.vi.08	Macro. willowae sp. n.	1	–
N005	Nicro. orbicollis	CAN, ON, Frontenac	44.447, -76.577	17.vi.08	Macro. willowae sp. n.	4	–
N006	Nicro. orbicollis	CAN, ON, Charleston Lake	44.500, -76.072	17.vi.08	Macro. willowae sp. n.	4	–
N007	Nicro. orbicollis	CAN, ON, Charleston Lake	44.500, -76.072	17.vi.08	Macro. willowae sp. n.	1	–
N008	Nicro. orbicollis	CAN, ON, Charleston Lake	44.500, -76.072	17.vi.08	Macro. willowae sp. n.	1	–
N011	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	2	–
N012	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	2	–
N013	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	5	–
N014	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	7	–
N015	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	5	–
N017	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	1	–
N018	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	2	–
N019	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	2	–
N020	Nicro. orbicollis	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	4	–
N021	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	1	–
N022	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	3	–
N023	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	2	–
N024	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	16.vi.08	Macro. willowae sp. n.	3	–
N026	Nicro. orbicollis	CAN, ON, Charleston Lake	44.500, -76.072	01.vii.08	Macro. willowae sp. n.	5	–
N028	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	3	–
N029	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	4	–
N031	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	2	–
N036	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	1	–
N037	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	3	–
N039	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	1	–
N040	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	8	–
N042	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	3	–
N043	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	3	–
N044	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	3	–
N046	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	1	–
N047	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	17	–
N048	Nicro. orbicollis	CAN, ON, Algonquin P.P. 1	45.902, -77.605	30.vi.08	Macro. willowae sp. n.	5	–
N051	Nicro. orbicollis	CAN, ON, Charleston Lake	44.500, -76.072	15.vii.08	Macro. willowae sp. n.	2	–
N052	Nicro. orbicollis	CAN, ON, Frontenac	44.447, -76.577	30.vii.08	Macro. willowae sp. n.	1	–
N055	Nicro. orbicollis	CAN, ON, Charleston Lake	44.500, -76.072	30.vii.08	Macro. willowae sp. n.	3	–
N057	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	29.vii.08	Macro. willowae sp. n.	1	–
N058	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	29.vii.08	Macro. willowae sp. n.	2	–
N060	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	29.vii.08	Macro. willowae sp. n.	2	–
N061	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	29.vii.08	Macro. willowae sp. n.	5	–
N062	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	29.vii.08	Macro. willowae sp. n.	3	–
N068	Nicro. defodiens	CAN, ON, Algonquin P.P. 1	45.902, -77.605	29.vii.08	Macro. willowae sp. n.	9	–
N069	Nicro. orbicollis	CAN, ON, Charleston Lake	44.500, -76.072	12.viii.08	Macro. willowae sp. n.	1	–
N075	Nicro. orbicollis	CAN, ON, Frontenac	44.447, -76.577	26.viii.08	Macro. willowae sp. n.	1	–
N081	Nicro. carolinus	USA, FL, Highlands Co, Lake Placid	27.181, -81.352	10.iii.2009	Macro. kaiju sp. n.	4	MF192750
N081	Nicro. carolinus	USA, FL, Highlands Co, Lake Placid	27.181, -81.352	10.iii.2009	Macro. willowae sp. n.	9	MF192743
N086	Nicro. orbicollis	CAN, ON, Windsor, Elgin St.	42.261, -83.057	18.vi.2009	Macro. willowae sp. n.	1	–
N088	Nicro. orbicollis	CAN, ON, Hwy 132, Dacre	45.369, -76.988	25.vi.2009	Macro. willowae sp. n.	10	–
N089	Nicro. orbicollis	CAN, ON, Carbine Rd.	45.33, -76.371	25.vi.2009	Macro. willowae sp. n.	9	–
N104	Nicro. defodiens	CAN, BC, Prince George, nr. UNBC	53.904, -122.783	12.vi.2009	Macro. willowae sp. n.	1	–
N110	Nicro. defodiens	CAN, BC, Prince George, nr. UNBC	53.904, -122.783	12.vi.2009	Macro. willowae sp. n.	1	–
N113	Nicro. defodiens	CAN, BC, Prince George, nr. UNBC	53.904, -122.783	12.vi.2009	Macro. willowae sp. n.	4	MF192748
N114	Nicro. orbicollis	CAN, PEI, Wellington, Route 2	46.452, -63.949	06.vii.2009	Macro. willowae sp. n.	8	–
N115	Nicro. orbicollis	CAN, PEI, Wellington, Route 2	46.452, -63.949	06.vii.2009	Macro. willowae sp. n.	7	–
N116	Nicro. orbicollis	CAN, PEI, Wellington, Route 2	46.452, -63.949	06.vii.2009	Macro. praedafimetorum	1	MF192754
N116	Nicro. orbicollis	CAN, PEI, Wellington, Route 2	46.452, -63.949	06.vii.2009	Macro. willowae sp. n.	9	MF192747
N136	Nicro. defodiens	CAN, BC, Prince George, nr. UNBC	53.904, -122.783	12.vi.2009	Macro. willowae sp. n.	1	MF192749
N139	Nicro. orbicollis	CAN, ON, Carbine Rd.	45.33, -76.371	10.vii.2009	Macro. willowae sp. n.	12	–
N143	Nicro. orbicollis	CAN, ON, Hamilton, Site 4-7		07.vii.2009	Macro. willowae sp. n.	100	MF192744
N145	Nicro. orbicollis	CAN, ON, Hamilton, Site 4-7		07.vii.2009	Macro. willowae sp. n.	8	–
N147	Nicro. orbicollis	CAN, ON, Hamilton, Site 4-7		07.vii.2009	Macro. willowae sp. n.	10	–
N153	Nicro. orbicollis	CAN, NS, Dartmouth, Wright’s Cove Rd.	44.694, -63.611	09.vii.2009	Macro. willowae sp. n.	4	–
N160	Nicro. orbicollis	CAN, ON, Waterloo	43.54, -80.211	07.vii.2009	Macro. willowae sp. n.	2	–
N161	Nicro. orbicollis	CAN, ON, Waterloo	43.54, -80.211	07.vii.2009	Macro. willowae sp. n.	8	–
N165	Nicro. orbicollis	CAN, ON, Carbine Rd.	45.33, -76.371	23.vii.2009	Macro. willowae sp. n.	17	–
N166	Nicro. orbicollis	CAN, ON, Carbine Rd.	45.33, -76.371	23.vii.2009	Macro. willowae sp. n.	8	–
N167	Nicro. orbicollis	CAN, NS, Glenmont, Black Hole Rd.	45.111, -64.296	17.vii.2009	Macro. willowae sp. n.	9	–
N168	Nicro. orbicollis	CAN, NS, Cold Brook, Hwy 101	45.079, -64.592	13.vii.2009	Macro. willowae sp. n.	10	MF192745
N169	Nicro. orbicollis	CAN, NS, Chipman	46.174, -65.899	08.vii.2009	Macro. willowae sp. n.	7	–
N174	Nicro. orbicollis	CAN, NS, Hantsport	45.099, -64.184	21.vii.2009	Macro. willowae sp. n.	6	–
N178	Nicro. orbicollis	CAN, NS, East River off Hwy 329	44.583, -64.164	10.viii.2009	Macro. willowae sp. n.	4	–
N180	Nicro. defodiens	CAN, NS, Debert, Industrial Park	45.428, -63.429	05.viii.2009	Macro. willowae sp. n.	3	–
N181	Nicro. orbicollis	CAN, ON, Carbine Rd.	45.33, -76.371	06.viii.2009	Macro. willowae sp. n.	8	MF192746
N185	Nicro. vespillo	GER, Mooswald Forest, nr. Freiburg	48.0, 7.85	vi.2009	Macro. nataliae	4	–
N186	Nicro. vespillo	GER, Mooswald Forest, nr. Freiburg	48.0, 7.85	vi.2009	Macro. nataliae	7	MF192752
N187	Nicro. vespillo	GER, Mooswald Forest, nr. Freiburg	48.0, 7.85	vi.2009	Macro. nataliae	1	–
N188	Nicro. vespillo	GER, Mooswald Forest, nr. Freiburg	48.0, 7.85	vi.2009	Macro. nataliae	5	MF192753
N191	Nicro. orbicollis	USA, CT, Bethany	41.462, -72.961	16.vii.2009	Macro. willowae sp. n.	1	–
N192	Nicro. orbicollis	USA, CT, Bethany	41.462, -72.961	14.viii.2009	Macro. willowae sp. n.	5	–
N216	Nicro. orbicollis	USA, NH, Durham	43.134, -70.926	07.vi.2009	Macro. willowae sp. n.	215	–
N218	Nicro. orbicollis	USA, NH, Durham	43.134, -70.926	07.vi.2009	Macro. willowae sp. n.	135	–
N222	Nicro. defodiens	USA, NH, Durham	43.134, -70.926	07.vi.2009	Macro. willowae sp. n.	30	–
N226	Nicro. orbicollis	USA, NH, Durham	43.134, -70.926	07.vi.2009	Macro. willowae sp. n.	30	–
N228	Nicro. orbicollis	USA, NH, Durham	43.134, -70.926	07.vi.2009	Macro. willowae sp. n.	97	–
N234	Nicro. marginatus	CAN, AB, Onefour	49.121, -110.47	17.vi.2003	Macro. pratum sp. n.	10	–
N235	Nicro. guttula	CAN, AB, Onefour	49.121, -110.47	17.vi.2003	Macro. pratum sp. n.	7	–
N235x	Nicro. marginatus	CAN, AB, Onefour	49.121, -110.47	04.vii.2002	Macro. pratum sp. n.	3	–
N236	Nicro. obscurus	CAN, AB, Onefour	49.121, -110.47	04.vii.2002	Macro. pratum sp. n.	5	–
N237	Nicro. marginatus	CAN, AB, Onefour	49.121, -110.47	17.vi.2003	Macro. pratum sp. n.	13	–
N238	Nicro. obscurus	CAN, AB, Onefour	49.121, -110.47	17.vi.2003	Macro. pratum sp. n.	5	–
N239	Nicro. obscurus	CAN, AB, Onefour	49.121, -110.47	17.vi.2003	Macro. pratum sp. n.	11	–
N240	Nicro. marginatus	CAN, AB, Onefour	49.121, -110.47	17.vi.2003	Macro. pratum sp. n.	13	–
N242	Nicro. hybridus	CAN, AB, Onefour	49.121, -110.47	18.vii.2002	Macro. pratum sp. n.	7	MF192751
N243	Nicro. guttula	CAN, AB, Onefour	49.121, -110.47	17.vi.2003	Macro. pratum sp. n.	7	–
N244	Nicro. guttula	CAN, AB, Onefour	49.121, -110.47	17.vi.2003	Macro. pratum sp. n.	7	–
N246	Nicro. hybridus	CAN, AB, Onefour	49.121, -110.47	18.vii.2002	Macro. pratum sp. n.	10	–
N274	Nicro. marginatus	CAN, AB, Onefour	49.121, -110.47	17.vi.2003	Macro. glaber	8	–
N275	Nicro. obscurus	CAN, AB, Onefour	49.121, -110.47	17.vi.2003	Macro. glaber	1	–
N295	Nicro. carolinus	USA, NE, Kearney Co.		05.vi.2009	Macro. kaiju sp. n.	1	–
N295	Nicro. carolinus	USA, NE, Kearney Co.		05.vi.2009	Macro. sp.	4	–
N298	Nicro. carolinus	USA, NE, Kearney Co.		05.vi.2009	Macro. kaiju sp. n.	2	–
N298	Nicro. carolinus	USA, NE, Kearney Co.		05.vi.2009	Macro. sp.	3	–
N303	Nicro. pustulatus	USA, NE, Kearney Co.		13.vii.2009	Macro. pratum sp. n.	3	–
N308	Nicro. marginatus	CAN, AB, Onefour	49.121, -110.47	17.vi.2003	Macro. pratum sp. n.	86	–
N329	Nicro. orbicollis	USA, NH		2009	Macro. willowae sp. n.	41	–
N330	Nicro. orbicollis	USA, NH		2009	Macro. willowae sp. n.	91	–
N331	Nicro. orbicollis	USA, CT, Bethany	41.462, -72.961	2009	Macro. willowae sp. n.	13	–
N332	Nicro. carolinus	USA, FL, Highlands Co, Lake Placid	27.181, -81.352	10.iii.2009	Macro. kaiju sp. n.	11	–
N333	Nicro. carolinus	USA, FL, Highlands Co, Lake Placid	27.181, -81.352	10.iii.2009	Macro. kaiju sp. n.	74	–
N334	Nicro. carolinus	USA, FL, Highlands Co, Lake Placid	27.181, -81.352	10.iii.2009	Macro. willowae sp. n.	1	–
N334	Nicro. carolinus	USA, FL, Highlands Co, Lake Placid	27.181, -81.352	10.iii.2009	Macro. kaiju sp. n.	26	–
N334	Nicro. carolinus	USA, FL, Highlands Co, Lake Placid	27.181, -81.352	10.iii.2009	Macro. sp.	1	–
N335	Nicro. carolinus	USA, FL, Highlands Co, Lake Placid	27.181, -81.352	10.iii.2009	Macro. kaiju sp. n.	45	–
N336	Nicro. marginatus	USA, NE, Kearney Co.		vi.2009	Macro. pratum sp. n.	9	–
N338	Nicro. defodiens	USA, NH, Durham	43.134, -70.926	07.vi.2009	Macro. willowae sp. n.	8	–
N339	Nicro. defodiens	USA, NH, Durham	43.134, -70.926	07.vi.2009	Macro. willowae sp. n.	163	–
N346	Nicro. orbicollis	CAN, ON, Hwy 132, Dacre	45.369, -76.988	06.viii.2009	Macro. willowae sp. n.	6	–
N347	Nicro. orbicollis	CAN, ON, Carbine Rd.	45.33, -76.371	06.viii.2009	Macro. willowae sp. n.	16	–

Pairwise distances were calculated using neighbour-joining (NJ) analyses with the Kimura-2-parameter (K2P) model in PAUP* v4.0b10 (Swofford 2003). Phylogenetic reco<span class="Disease">nstructio<al">span class="Disease">ns of the COI dataset was performed using Bayesian inference (BI) in MrBayes v3.2.6 (Huelsenbeck and Ronquist 2001; Ronquist and Huelsenbeck 2003). Each specimen in the phylogeny is labeled with the mite species and the beetle number, followed by the host species and abbreviated state, province or country (Fig. 14).

Figure 14.

Majority rule consensus tree of 19502 trees generated by Bayesian MCMC analysis (10 million generations) of 689bp fragment of COI from 14 ingroup specimens representing six species, and two outgroup specimens representing two species, posterior probabilities >50% shown above branches.

MrModeltest v2.3 (Nylander 2004) was used to determine the best-fit model of molecular evolution for each molecular marker, which was determined to be GTR+I+G. Bayesian analysis was performed in MrBayes using the Markov Chain Monte Carlo (MCMC) method, two independent ru<span class="Disease">ns, with nucmodel = 4by4, <al">span class="Disease">Nst = 6, rates = invgamma, samplefreq = 1000, four chains = one cold and three heated. The COI dataset ran for 10 million generations, producing 19502 trees after a burn-in of 250 trees. The remaining trees in Mesquite, excluding the burn-in, were used to generate a majority-rule consensus tree displaying the posterior probability supports for each node. Bayesian analyses were performed using the on-line Computational Biology Service Unit at Cornell University, and at the Cyberinfrastructure for Phylogenetic Research (CIPRES) portal (Miller et al. 2010).

Results and discussion

Family Vitzthum, 1930

Subfamily

Latreille, 1829

Type species.

Hermann, 1804 (= Scopoli, 1772), by original designation.

sp. n.

http://zoobank.org/1085F03B-0DC9-4762-91E4-6FA1770E6965

Figs 1 , 2 , 3 , 4 , 13A

Figure 1.

Female sp. n. A dorsal idiosoma B seta J5.

Figure 2.

Female sp. n. ventral idiosoma including coxae.

Figure 3.

Female sp. n. A subcapitulum and palp, ventral aspect B chelicera, antiaxial aspect C epistome D tritosternum.

Figure 4.

Female sp. n. legs I–IV, coxae omitted; leg I anterolateral, II posterolateral, III dorsal, IV anterolateral.

Figure 13.

Sternal shield A sp. n. B sp. n. C sp. n.

Material examined.

Holotype: female (CNC829414) on (N088, female) collected near Dacre, Ontario, Canada (), 25.vi.2009, coll: W. Knee.

Paratypes (26): Nine females (CNC829415–829423) with the same collection information as the holotype; 15 females (CNC829424–829438) on (N222, female), Durham, New Hampshire, USA (), 07.vi.2009, coll: W. Knee & M. Scott; female (CNC829439) on (N136, female), Prince George, near University of Northern <span class="Disease">British Columbia campus, British Columbia, Canada (), 12.vi.2009, coll: W. Knee & R. Dawson; female (CNC829440) on (N143, male), Hamilton, Ontario, Canada, 7.vii.2009, coll: W. Knee.

1241 mites examined from <span class="Disease">British Columbia, Nova Scotia, Ontario, Prince Edward Island, Connecticut, Florida, and New Hampshire on , and (Table 1).

Diagnosis female.

As for (see Hyatt and Emberson 1988). All dorsal and ventral setae smooth and spinose, except J5 barbed and slightly shorter than Z5. Seta j1 simple with rounded tip, j1 slightly longer than z1. Dorsal hexagonal setae (j5, z5, j6) nearly as long as marginal and submarginal setae (R and UR). Dorsal shield with moderate reticulatio<span class="Disease">ns throughout, except smooth in dorsal hexagonal area and between j4 setae, without well-defined procurved line, sigillary rami absent. Sternal shield more than twice as wide as long, punctures small, posterior margin concave. Well defined linea media tra<al">span class="Disease">nsversa (l.m.t.) and linea oblique anteriores (l.o.a.), l.o.a. contacts l.m.t. (l.arc.) well defined and contacts l.o.a. (l.ang.) and linea oblique posteriore (l.o.p.) well defined laterally but faint medially. (a.p.l.) well defined, but area (a.p.p.) not well defined. Ventrianal shield longer than wide (ratio 1.3). Arthrodial brush as long as movable digit. Genu IV with six setae. Femur IV setae ad2, pd1 prominent spikes with flattened forked tip.

Description female.

(Fig. 1). Dorsal shield 548 (526–572) long and 357 (344–372) (n=8) wide (level with r3), with 28 pairs of setae, all setae simple and spinose except J5 is barbed. Seta J5 16 (15–18) shorter than Z5 24 (22–27). Seta j1 simple with rounded tip, j1 18 (16–20) slightly longer than z1 16 (12–19). Marginal and submarginal setae simple, slightly longer 24 (23–25) than dorsal hexagonal setae 20 (15–22). Dorsal shield with moderate reticulatio<span class="Disease">ns throughout, except smooth in dorsal hexagonal area and between j4 setae, shield without well-defined procurved line, sigillary rami absent, and posterolateral margi<al">span class="Disease">ns narrowed slightly. Shield with 22 pairs of pore-like structures, of which six are secretory glands and 16 are non-secretory poroids.

Female sp. n. A <span class="Disease">dorsal idiosoma B seta J5.

(Figs 2, 13A). Sternal shield more than twice as wide as long, medial length 91 (88–96), maximum width 213 (192–221) level with a.p.l., and minimum width 117 (115–124) posterior of st1. Sternal shield punctures small, posterior margin concave. Setae st1–3 38 (33–43) simple and spinose, and two pairs of lyrifissures (iv1, iv2) on sternal shield. <span class="Species">Pear-shaped metasternal shields well separated from sternal shield margin bearing lyrifissure iv3 anteriorly and al">spinose seta st4 33 (30–35) posteriorly. Well defined l.m.t. and l.o.a., l.o.a. contacts l.m.t. Well defined l.arc. contacts l.o.a., l.ang. and l.o.p. well defined laterally but faint medially. Well defined a.p.l., a.p.p. not well defined. Genital shield length 149 (141–161), width 113 (104–120) level with st5. Genital shield truncate posteriorly and hyaline margin rounded anteriorly, al">spinose seta st5 32 (30–34) on shield, pair of lyrifissures iv5 off shield near posterior margin. Tra<al">span class="Disease">nsverse line on genital shield well defined laterally and faint medially, small punctures along transverse line. Peritrematal shield narrow, fused to dorsal shield near r3, peritreme extends beyond posterior margin of coxa I, two poroids (id3, id7) and one gland (gd3) on the shield. Ventrianal shield longer than wide (ratio 1.3); length 198 (187–204), width 153 (144–168) level with JV2. Ventrianal shield bearing several faint transverse lines, three pairs of simple spinose preanal setae JV1–JV3 26 (21–30), spinose paranal (pan) 27 (25–30) and postanal (pon) 18 (16–20) setae, narrow cribrum and a pair of glands (gv3) on shield margin posterior of the anal opening. Ventral opisthosomal setae in soft integument simple and spinose, ZV1 21 (17–29), ZV2 26 (22–29) as long or nearly as long as Jv setae. Two pairs of glands (gv2 and unknown paired-pore) and four pairs of poroids (ivo, ivp) in opisthosomal soft integument.

Female sp. n. <span class="Disease">ventral idiosoma including coxae.

(Fig. 3). Basis capitulum medial length excluding internal malae 114 (106–122), width 150 (143–157) posterior to pc. Subcapitular setae simple: h1 38 (28–48), <span class="Chemical">h2 16 (13–19), <al">span class="Chemical">h3 53 (44–67), and pc 19 (16–21). Palp chaetotaxy normal for genus (2–5–6–14–15), palp apotele three-tined, al setae on trochanter, femur and genu slightly spatulate. Corniculi pointed, length along lateral margin 42 (37–51), internal malae slender and smooth. Epistome tripartite with bifid central element bearing small fringe medially, lateral elements broad and flag-like distally, epistomatic margin finely serrate. Subcapitulum with seven rows, six of which have deutosternal denticles; the anterior most row with few (four) denticles laterally, and the second anterior most row with paired ridges without any denticles. Chelicerae robust, length of second cheliceral segment including fixed digit 140 (134–146), and movable digit 54 (52–58). Fixed digit bidentate with one large and one small tooth, moveable digit with bidentate tooth. Pilus dentilis and dorsal seta on fixed digit simple spikes, fixed digit with lyrifissure on each paraxial and antiaxial faces. Movable digit with narrow fringed arthrodial corona, and plumose arthrodial brush (50) almost as long as movable digit.

(Fig. 4). Excluding ambulacra, lengths of <span class="Disease">leg I 420 (409–430), <al">span class="Disease">leg II 383 (357–402), leg III 338 (315–349), and leg IV 482 (469–489). As in all , ambulacra only present on legs II–IV, claws II–IV well developed. Pair of glands (gc) on coxa I. Setation of legs I–IV normal for : coxae 2–2–2–1; trochanters 5–5–5–5; femora I (2–3/1,2/3–2) (as al–ad/av, pd/pv–pl), II (2–3/1,2/2–1), III (1–2/0,1/1–1), IV (1–2/1,1/0–1); genua I, II (2–3/1,2/1–2), III (1–2/1,2/0–1), IV (1–2/1,2/0–0); tibiae I (2–3/2,2/1–2), II (2–2/1,2/1–2), III, IV (1–1/1,2/1–1); tarsus I 20 setae plus numerous tapered setae distally, tarsi II–IV 18. Most leg setae simple, setiform, femur II ad1, III pd1, IV ad2, pd1, and genu II ad3 prominent spike setae with flattened forked tip with two to four tines that can appear as a single tapered point viewed laterally. Tarsus II with four large distal spike setae with thickened conical base and rounded tip. Tarsi III, IV with four and three, respectively, distal setae PageBreakwith wide base and flexible filamentous tip. Genu and tibia IV with paired slight ridge anterolateral and posterolateral.

Female sp. n. <span class="Disease">legs I–IV, coxae omitted; <al">span class="Disease">leg I anterolateral, II posterolateral, III dorsal, IV anterolateral.

Male and immatures.

Unknown.

Etymology.

This species is named after my daughter Willow Knee. May it i<span class="Disease">nspire her to notice the little creatures as well as the big.

Remarks.

sp. n. is most similar to (Berlese), Krantz and Whitaker, and sp. n. is frequently found in litter, manure and compost worldwide, feeding on nematodes and eggs of i<span class="Disease">nsects (Krantz and Whitaker 1988). has been reported from small <al">span class="Species">mammals, and female mites are often phoretic associates of dung beetles (Filipponi and Pegazzano 1963, Krantz and Whitaker 1988). Female sp. n. differs from that of in the shape of the ventrianal shield, shape of the sternal shield, and length of the arthrodial brush. The anterior margin of the ventrianal shield is more truncated, and the widest part of the shield near JV2 is more angular for than it is for sp. n. The posterior margin of the sternal shield PageBreakPageBreakis more concave for sp. n. The arthrodial brush is almost as long as the movable digit for sp. n., and for the brush is approximately half as long as the movable digit. Comparisons were made using the species description for and slide mounted material deposited in the CNC.

was described from the nest of a <span class="Species">mouse, in Maryland and the nest of the <al">span class="Species">eastern woodrat in Florida, USA, and this species is also phoretic on coprophilous beetles (Krantz and Whitaker 1988). Female sp. n. differs from that of in having marginal or submarginal setae slightly longer than dorsal hexagonal setae, posterior margin of sternal shield more concave, pon seta smooth not weakly pilose, j1 only slightly longer than z1 not 1.5 times as long, and J5 slightly shorter than Z5 not half as long as Z5. Comparisons were made using the species description for and examination of the holotype specimen loaned from the National Museum of Natural History, Smithsonian Institution.

Female sp. n. differs from that of sp. n. in having marginal and submarginal setae slightly longer than dorsal hexagonal setae. Genu and <span class="Disease">tibia IV with slight ridge on anterolateral and posterolateral surfaces in al">sp. n., while al">sp. n. only has a ridge on the posterolateral surface. Seta J5 is slightly shorter than Z5 and more al">spinose in al">sp. n., J5 is less than half as long as Z5 in al">sp. n. Punctures on the sternal shield are smaller and less prominent in al">sp. n. than in al">sp. n. The ventrianal shield is longer than wide for both al">species, but the shield is slightly narrower in al">sp. n., ratio of 1.4 compared to 1.3 for al">sp. n.

In Krantz and Whitaker (1988), Dr. W. Yoder provided a short diagnosis and partial illustratio<span class="Disease">ns of the female and male of an undescribed and unnamed al">species collected from beetles and three <al">span class="Species">mammal species (, American red squirrel in Michigan; , eastern chipmunk in Maryland; and , meadow jumping mouse in Prince Edward Island). This undescribed species was a common associate of beetles, but it was also found frequently enough on live rodents to suggest an association with small mammals (Krantz and Whitaker 1988). Dr. W. Yoder reportedly intended to formally describe and illustrate this new species of ; however, to date this species has not been described. sp. n. is likely the same species that Dr. W. Yoder was intending to describe. Over several years, repeated attempts were made to contact Dr. W. Yoder about the status of the description, but contact was unsuccessful.

http://zoobank.org/036A8E8D-9669-4524-89B4-4395BC71385A

Figs 5 , 6 , 7 , 8 , 13B

Figure 5.

Female sp. n. A dorsal idiosoma B seta J5.

Figure 6.

Female sp. n. ventral idiosoma including coxae.

Figure 7.

Female sp. n. A subcapitulum and palp, ventral aspect B chelicera, antiaxial aspect C epistome D tritosternum.

Figure 8.

Female sp. n. legs I–IV, coxae omitted; leg I anterolateral, II posterolateral, III posterolateral, IV femur dorsal, genu, tibia, tarsus anterolateral.

Holotype: female (CNC829441) on (N336, female) collected in Kearney Co., Nebraska, USA, vi.2009, coll: W. Knee & W. Hoback.

Paratypes (11): eight females (CNC829442–829449) with the same collection information as the holotype; two females (CNC829450, 829451) on (N242, female), Onefour, Alberta, Canada, 18.vii.2002, coll: W. Knee & D. Joh<span class="Disease">nson; female (CNC829452) on (N235, female), Onefour, Alberta, 17.vi.2003, coll: W. Knee & D. Joh<al">span class="Disease">nson.

184 mites examined from Alberta and Nebraska on , , , , and (Table 1).

All dorsal and ventral setae smooth and spinose, except J5 barbed and much shorter than Z5. Seta j1 simple with rounded tip, j1 slightly longer than z1. Dorsal hexagonal setae slightly longer than marginal and submarginal setae. Dorsal shield with moderate reticulatio<span class="Disease">ns throughout, except smooth in dorsal hexagonal area and between j4 setae, without well-defined procurved line, sigillary rami absent. Sternal shield more than twice as wide as long, punctures moderate size, posterior margin concave. Well defined l.m.t. and l.o.a.; l.o.a. contacts l.m.t. Well defined l.arc. contacts l.o.a., l.ang. and l.o.p. well defined laterally but faint medially. Well defined a.p.l., but a.p.p. not well defined. Ventrianal shield longer than wide (ratio 1.4). Arthrodial brush as long as movable digit. Genu IV with six setae. Femur IV setae ad2, pd1 prominent al">spikes with flattened forked tip.

(Fig. 5). Dorsal shield 520 (469–547) long and 358 (323–379) (n=8) wide (level with r3), with 28 pairs of setae, all setae simple and spinose except J5 barbed. Seta J5 9 (8–10) half as long as Z5 22 (18–23). Seta j1 simple with rounded tip, j1 20 (19–21) longer than z1 16 (13–18). Marginal and submarginal setae simple, shorter (19) than dorsal hexagonal setae 25 (24–28). Dorsal shield with moderate reticulatio<span class="Disease">ns throughout, except smooth in dorsal hexagonal area and between j4 setae, shield without well-defined procurved line, sigillary rami absent, and posterolateral margi<al">span class="Disease">ns narrowed slightly. Shield with 22 pairs of pore-like structures, of which six are secretory glands and 16 are non-secretory poroids.

Female sp. n. A <span class="Disease">dorsal idiosoma B seta J5.

(Figs 6, 13B). Sternal shield more than twice as wide as long, medial length 92 (86–99), maximum width 217 (196–224) level with a.p.l., and minimum width 114 (110–119) posterior of st1. Sternal shield punctures moderate size, posterior margin concave. Setae st1–3 40 (35–45) simple and spinose, and two pairs of lyrifissures (iv1, iv2) on sternal shield. <span class="Species">Pear-shaped metasternal shields well separated from sternal shield margin bearing lyrifissure iv3 anteriorly and al">spinose seta st4 35 (33–39) posteriorly. Well defined l.m.t. and l.o.a., l.o.a. contacts l.m.t. Well defined l.arc. contacts l.o.a., l.ang. and l.o.p. well defined laterally but faint medially. Well defined a.p.l., a.p.p. not well defined. Genital shield length 151 (138–158), width 112 (99–122) level with st5. Genital shield truncate posteriorly and hyaline margin rounded anteriorly, al">spinose seta st5 32 (30–34) on shield, pair of lyrifissures iv5 off shield near posterior margin. Tra<al">span class="Disease">nsverse line on genital shield well defined laterally and faint medially, small punctures along transverse line. Peritrematal shield narrow, fused to dorsal shield near r3, peritreme extends beyond posterior margin of coxa I, two poroids (id3, id7) and one gland (gd3) on the shield. Ventrianal shield longer than wide (ratio 1.4); length 186 (168–194), width 135 (122–146) level with JV2. Ventrianal PageBreakPageBreakPageBreakshield bearing several faint transverse lines, three pairs of simple spinose preanal setae JV1–JV3 26 (21–29), spinose pan 25 (22–28) and pon 18 (15–19), narrow cribrum and a pair of glands (gv3) on shield margin posterior of the anal opening. Ventral opisthosomal setae in soft integument simple and spinose, ZV1 17 (13–23), ZV2 25 (21–27) as long or nearly as long as Jv setae. Two pairs of glands (gv2 and unknown paired-pore) and four pairs of poroids (ivo, ivp) in opisthosomal soft integument.

Female sp. n. <span class="Disease">ventral idiosoma including coxae.

(Fig. 7). Basis capitulum medial length excluding internal malae 115 (113–116), width 139 (134–143) posterior to pc. Subcapitular setae simple: h1 44 (41–48), <span class="Chemical">h2 16 (13–18), <al">span class="Chemical">h3 57 (51–62), and pc 20 (19–23). Palp chaetotaxy normal for genus (2–5–6–14–15), palp apotele three-tined, al setae on trochanter, femur and genu slightly spatulate. Corniculi pointed, maximum length 39 (35–45), internal malae slender and smooth. Epistome tripartite with bifid central element bearing small fringe medially, lateral elements broad and flag-like distally, epistomatic margin finely serrate. Subcapitulum with seven rows: six rows have deutosternal denticles, the anterior most, and two posterior most rows with few (four) denticles laterally; the second anterior most row with paired ridges without any denticles. Chelicerae robust, length of second cheliceral segment including fixed digit 135 (122–141), and movable digit 49 (45–52). Fixed digit bidentate with one large and one small tooth, movable digit with a bidentate tooth flanked by a small tooth distally. Pilus dentilis and dorsal seta on PageBreakfixed digit simple spikes, fixed digit with lyrifissure on each paraxial and antiaxial faces. Movable digit with narrow fringed arthrodial corona, and plumose arthrodial brush (47) almost as long as movable digit.

(Fig. 8). Excluding ambulacra, lengths of <span class="Disease">leg I 418 (409–432), <al">span class="Disease">leg II 390 (362–438), leg III 340 (319–358), and leg IV 474 (463–490). As in all PageBreak ambulacra only present on legs II–IV, claws II–IV well developed. Pair of glands (gc) on coxa I. Setation of legs I–IV normal for : coxae 2–2–2–1; trochanters 5–5–5–5; femora I (2–3/1,2/3–2), II (2–3/1,2/2–1), III (1–2/0,1/1–1), IV (1–2/1,1/0–1); genua I, II (2–3/1,2/1–2), III (1–2/1,2/0–1), IV (1–2/1,2/0–0); tibiae I (2–3/2,2/1–2), II (2–2/1,2/1–2), III, IV (1–1/1,2/1–1); tarsus I 20 setae plus numerous tapered setae dorsoterminally, tarsi II–IV 18. Most leg setae simple, setiform, femur II ad1, III pd1, IV ad2, pd1, and genu II ad3 prominent spike setae with flattened forked tip with two to four tines that can appear as a single tapered point viewed laterally. Tarsus II with four large distal, and one ventral, spike setae with thickened conical base and rounded tip. Tarsi III, IV with four distal spike setae with wide base and flexible filamentous tip. Genu and tibia IV with slight ridge posterolateral.

Female sp. n. <span class="Disease">legs I–IV, coxae omitted; <al">span class="Disease">leg I anterolateral, II posterolateral, III posterolateral, IV femur dorsal, genu, tibia, tarsus anterolateral.

Unknown.

Pratum (Latin neuter noun) mea<span class="Disease">ns “meadow”. This al">species was only collected in Kearney County, Nebraska and Onefour, Alberta, which are in the prairies.

The female of sp. n. is most similar to those of sp. n., , Berlese, and Berlese. sp. n. differs from sp. n. as outlined in the sp. n. description.

Female sp. n. differs from that of in having larger more prominent punctures on the sternal shield, the posterior margin of the sternal shield is more concave, pon seta is smooth not weakly pilose, j1 is only slightly longer than z1 not 1.5 times as long, J5 is shorter and broader for sp. n. (9) than for (13), and the ventrianal shield is narrower, length to width ratio of 1.4 for sp. n. and 1.2 for . Measurements were made examining holotype specimen loaned from the National Museum of Natural History, Smithsonian I<span class="Disease">nstitution.

and are associated with coprophilous beetles (Krantz 1988). Female sp. n. differs from those of and in having larger more prominent punctures and tra<span class="Disease">nsverse lines on the sternal shield, the posterior margin of the sternal shield more concave, arthrodial brush nearly as long as movable digit and not half or three quarters as long as movable digit, setae ad2 and pd1 on femur IV are large al">spike-like setae with flattened forked tips with two to four tines, ventrianal shield tapers relatively more towards posterior starting anterior of pan setae, and ventrianal shield is narrower, length to width ratio of 1.4 for al">sp. n. and 1.2 for and . Measurements were made examining and voucher material from the Oregon State University Arthropod Collection.

http://zoobank.org/1AFFAB07-D38<span class="CellLine">F-429E-9F19-A76E2254787E

Figs 9 , 10 , 11 , 12 , 13C

Figure 9.

Female sp. n. A dorsal idiosoma B seta J5.

Figure 10.

Female sp. n. ventral idiosoma including coxae.

Figure 11.

Female sp. n. A subcapitulum and palp, ventral aspect B chelicera, antiaxial aspect C epistome D tritosternum.

Figure 12.

Female sp. n. legs I–IV, coxae omitted; leg I anterolateral, II posterolateral, III and IV dorsal.

Holotype: female (CNC829453) on (N333, male) collected Highlands Co. Lake Placid, Florida, USA (), 10.iii.2009, coll: W. Knee & S. Peck.

Paratypes (28): 14 females (CNC CNC829454–829467) with the same collection information as the holotype; 13 females (CNC829468–829480) on (N334, female), Highlands Co. Lake Placid, Florida, USA (), 10.iii.2009, coll: W. Knee & S. Peck; female (CNC829481) on (N081), Highlands Co. Lake Placid, Florida, USA (), 10.iii.2009, coll: W. Knee & S. Peck.

134 mites examined from Florida and Nebraska on (Table 1).

Dorsal setae smooth and spinose, except r3, r4, s6, z6, S1–S5, Z1–Z5, J2 barbed distally, J5 barbed, marginal and submarginal setae barbed distally. Seta j1 smooth, spike, tapered distally with rounded tip, slightly longer than z1. Seta J5 much shorter than Z5. Dorsal hexagonal setae as long as marginal and submarginal setae. Dorsal shield smooth medially with faint reticulatio<span class="Disease">ns near shield margi<al">span class="Disease">ns, shield tapers from humeral region to posterior margin. Dorsal shield without well-defined procurved line, sigillary rami absent. Setae on sternal, genital and ventrianal shields, and ZV1 smooth and spinose, other ventral setae in soft integument barbed distally. Sternal shield wider than long, punctures small, posterior margin slightly concave. Well defined l.m.t. and l.o.a.; l.o.a. contacts l.m.t. Well defined l.arc. contacts l.o.a., l.ang. and l.o.p. well defined laterally but faint medially. Well defined a.p.l., but a.p.p. not well defined. Ventrianal shield longer than wide (ratio 1.5), pon longer than pan, pon slightly spatulate. Arthrodial brush nearly as long as movable digit. Genu IV with six setae.

(Fig. 9). Dorsal shield 736 (647–812) long and 484 (421–547) (n=8) wide (level with r3), with 28 pairs of setae. Dorsal setae smooth and spinose, except r3, r4, s6, z6, S1–S5, Z1–Z5, J2 barbed distally, J5 barbed, marginal and submarginal setae barbed distally. Seta J5 22 (19–24) less than half as long as Z5 75 (65–83). Seta j1 29 (25–34) smooth, spike, tapered distally with rounded tip, slightly longer than z1 24 (20–32). Dorsal hexagonal setae 61 (50–71) smooth and spinose, as long as distally barbed marginal and submarginal setae (61). Dorsal shield smooth medially with faint reticulatio<span class="Disease">ns near shield margi<al">span class="Disease">ns, shield tapers from humeral region to posterior margin. Dorsal shield without well-defined procurved line, sigillary rami absent. Dorsal shield fused to peritrematal shield near r3 and anterior margin of shield wraps around onto ventral surface, j1 on slight projection and typically on the venter, and z1 occasionally expressed ventrally. Shield with 22 pairs of pore-like structures, of which six are secretory glands and 16 are non-secretory poroids.

Female sp. n. A <span class="Disease">dorsal idiosoma B seta J5.

(Figs 10, 13C). Sternal shield wider than long, medial length 155 (145–163), maximum width 260 (224–288) level with a.p.l., and minimum width 115 (103–121) posterior of st1. Sternal shield punctures small, posterior margin slightly concave. Setae st1–3 75 (61–92) simple and spinose, and two pairs of lyrifissures (iv1, iv2) on sternal shield. <span class="Species">Pear-shaped metasternal shields well separated from sternal shield margin bearing lyrifissure iv3 anteriorly and al">spinose seta st4 80 (73–89) posteriorly. Well defined l.m.t. and l.o.a.; l.o.a. contacts l.m.t. Well defined l.arc. contacts l.o.a., l.ang. and l.o.p. well defined laterally but faint medially. Well defined a.p.l., a.p.p. not well defined. Genital shield length 196 (172–226), width 123 (109–141) PageBreaklevel with st5. Genital shield truncate posteriorly and hyaline margin rounded anteriorly, al">spinose seta st5 70 (65–74) on shield, pair of lyrifissures iv5 off shield near posterior margin. Tra<al">span class="Disease">nsverse line on genital shield well defined, and without punctures. Peritrematal shield narrow, fused to dorsal shield near r3, peritreme extends beyond PageBreakPageBreakposterior margin of coxa I, two poroids (id3, id7) and one gland (gd3) on the shield. Ventrianal shield longer than wide (ratio 1.5); length 230 (202–250), width 149 (135–160) anterior to JV2. Ventrianal shield bearing several faint transverse lines, three pairs of simple spinose preanal setae JV1–JV3 65 (64–75), spinose pan 37 (26–43) shorter than slightly spatulate pon 47 (40–54), narrow cribrum and a pair of glands (gv3) on shield margin posterior of the anal opening. Seta ZV1 55 (48–60) is simple, all other ventral opisthosomal setae in soft integument barbed distally, ZV1 and ZV2 67 (58–75) as long or nearly as long as Jv setae. Two pairs of glands (gv2 and unknown paired-pore) and four pairs of poroids (ivo, ivp) in opisthosomal soft integument.

Female sp. n. <span class="Disease">ventral idiosoma including coxae.

(Fig. 11). Basis capitulum medial length excluding internal malae 159 (152–169), width 158 (153–170) posterior to pc. Subcapitular setae simple: h1 66 (61–70), <span class="Chemical">h2 19 (17–21), <al">span class="Chemical">h3 92 (85–98), and pc 25 (24–28). Palp chaetotaxy normal for genus (2–5–6–14–15), palp apotele three-tined, al setae on trochanter, femur and genu slightly spatulate. Corniculi pointed, maximum length 50 (45–60), internal malae thick and bristled. Epistome tripartite with bifid central element bearing small fringe medially, lateral elements broad and flag-like distally with irregular barbs, epistomatic margin finely serrate. Subcapitulum with seven rows, six of which have deutosternal denticles; the anterior most row with few (four) denticles laterally, and the second anterior most row with paired ridges without any denticles. Chelicerae robust, length of second cheliceral segment including fixed digit 184 (173–198), and movable digit 64 (60–67). Fixed digit bidentate with one large and one small tooth, movable digit with a bidentate tooth flanked by a small tooth distally. Pilus dentilis and dorsal seta on fixed digit simple spike, fixed digit with lyrifissure on each paraxial and antiaxial faces. Movable digit with narrow fringed arthrodial corona, and plumose arthrodial brush (57) almost as long as movable digit.

(Fig. 12). Excluding ambulacra, lengths of <span class="Disease">leg I 512 (488–527), <al">span class="Disease">leg II 576 (525–622), leg III 482 (436–528), and leg IV 690 (628–737). Ambulacra only present on legs II–IV, claws II–IV well developed. Slight ridge on femur II anterolateral, not always easily visible. Slight ridge on femur IV dorsal, genu and tibia IV posterolateral. Pair of glands (gc) on coxa I. Setation of legs I–IV normal for : coxae 2–2–2–1; trochanters 5–5–5–5; femora I (2–3/1,2/3–2), II (2–3/1,2/2–1), III (1–2/0,1/1–1), IV (1–2/1,1/0–1); genua I, II (2–3/1,2/1–2), III (1–2/1,2/0–1), IV (1–2/1,2/0–0); tibiae I (2–3/2,2/1–2), II (2–2/1,2/1–2), III, IV (1–1/1,2/1–1); tarsus I 20 setae plus numerous tapered setae dorsoterminally, tarsi II–IV 18. Setae on leg I are setiform and simple, setae on legs II–IV variable, most are setiform, others are variously modified. Femur II ad1, genu II ad3, and trochanter III with a prominent spike setae with flattened forked tip with two to four tines that can appear as a single tapered point viewed laterally. Tarsus II with 13 thick conical spike setae with either a rounded or filamentous tip; filamentous tip fragile and easily broken. Tarsi III, IV with four distal spike setae with wide base and flexible filamentous tip, tip easily broken. Long setae with small spatulate tip with or without barbs on femur IV ad1, ad2, pd1, genu IV al, ad1, pd1, pd2, tibia IV al, pl, and three on tarsus IV. Setae with small spatulate tip appear pointed when viewed laterally.

Female sp. n. <span class="Disease">legs I–IV, coxae omitted; <al">span class="Disease">leg I anterolateral, II posterolateral, III and IV dorsal.

Male and immatures

. Unknown.

Kaiju, 怪獣, from Japanese mea<span class="Disease">ns strange beast, and refers to giant mo<al">span class="Disease">nsters such as Godzilla or Mothra. Female sp. n. is relatively unique morphologically when compared to other species associated with beetles, it is relatively large, has a unique dorsal shield shape, and bears numerous setae with distinct forms.

Female sp. n. is different from that of any other described species; however, it does fit the generic description (see HyPageBreakPageBreakatt and Emberson 1988). Female sp. n. has two character states that are irregular for but somewhat similar to () species: the anterior margin of the dorsal shield wraps around onto the ventral surface such that j1 is on a slight projection on the venter; and a slight ridge is present on femur II. species are characterised in part by females having a spur on femur II, and j1 being on a tuberculate anterior extension of the dorsal shield but not wrapped around onto the venter. al">sp. n. differs from that of al">species in having weak ornamentation on the dorsal shield, its lateral regio<al">span class="Disease">ns without a series of depressions; j1 is smooth and not pectinate, j1 is on a slight projection and not a prominent tuberculate extension; sternal shield weakly ornamented and without strong median ridge; and metasternal shields small and always free of endopodal shields.

Phylogenetics

COI was amplified from 14 specime<span class="Disease">ns representing six al">species, with 689 characters in total, 430 co<al">span class="Disease">nstant, 39 parsimony-uninformative, 220 parsimony-informative. NJ analysis (K2P) was performed on 14 ingroup specimens. Average intraspecific pairwise distance was low (1.2% ±1.4), and the maximum intraspecific divergence observed was for sp. n. (2.8%). The higher than average intraspecific divergence for sp. n. was due to the divergence between mites from different host species. Pairwise distance between sp. n. mites from and those from and was higher than average (5.2% ±0.1), while divergence among mites from and mites from and was low, 0.1 and 0.6% respectively. Mean interspecific divergence was high (20% ±2.8), and the maximum divergence observed was between and (25%). The range of intra- (0.3–2.8%) and interspecific (15–25%) pairwise divergence did not overlap.

The majority rule co<span class="Disease">nse<al">span class="Disease">nsus tree from the BI analysis of COI was well supported, with all nodes having high posterior probabilities, and eight nodes with 100% support (Fig. 14). sp. n. was divided into two well supported clades, one with mites from and the other with mites from and . sp. n. did not appear to diverge based on geographic location (Fig. 14). sp. n. mites from and those from and were morphologically indistinguishable, despite the higher than average intraspecific divergence between these two well supported clades. The phylogenetic relationships between species, and the genetic structure of these newly described species, requires further analysis with better taxon sampling, specimens from more host species and localities, and additional molecular markers.

Majority rule co<span class="Disease">nse<al">span class="Disease">nsus tree of 19502 trees generated by Bayesian MCMC analysis (10 million generations) of 689bp fragment of COI from 14 ingroup specimens representing six species, and two outgroup specimens representing two species, posterior probabilities >50% shown above branches.

Distribution and biology

Seven species of macrochelids were collected from 112 beetles representing nine species from three countries (Canada, USA, Germany) and 10 provinces or states (Table 1). was associated with the most species (3), three host species had only two macrochelid species, and five had only one macrochelid species. Mites were usually found under the elytra, either clasping onto the integument near the prospiracle (68%) or on the ventral surface of the elytra (22%), and sometimes they were on the coxae (10%). Mites attached to the outer surface of the beetle could have been dislodged into the preservative.

A total of 1659 mites were collected from 112 beetles, sp. n. (1268 mites on 86 beetles) was the most abundant, second most abundant was sp. n. (196 mites on 15 beetles), sp. n. (163 mites on 7 beetles) was the third most abundant, and the four other species of collected were at PageBreaklow abundances (32 mites total on 10 beetles) (Table 1). sp. n. was collected from five host species and had the greatest host range of all species collected. sp. n. collected from three host species had the second broadest PageBreakhost range. , , and sp. n. were each collected from a single host species.

The species with the greatest geographic range was sp. n., collected from 22 sites, across seven provinces/states in Canada and USA. sp. n. was collected from a single site in Alberta (Canada) and from another site in Nebraska (USA). sp. n. was collected from one site in Florida and another site in Nebraska, USA. The four other macrochelid species collected were each found in a single locale (Table 1).

COI sequences generated in this study were compared agai<span class="Disease">nst those on GenBank and BOLD, and al">sp. n. was the only al">species to have high level (100%) matches on BOLD. These matching sequences belonged to generic level identified al">specime<al">span class="Disease">ns from Alberta, Ontario, Saskatchewan, Nova Scotia, and Florida. The species briefly diagnosed by Dr. W. Yoder was collected from three species of rodents in Maryland, Michigan and Prince Edward Island. Combined together, results from this study, BOLD and Dr. W. Yoder’s findings, the geographic distribution of sp. n. may cover 11 provinces or states in Canada and USA.