The oldest continuous association between astigmatid mites and termites preserved in Cretaceous amber reveals the evolutionary significance of phoresy

The oldest continuous association between astigmatid mites and termites preserved in Cretaceous amber reveals the evolutionary significance of phoresy | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article The oldest continuous association between astigmatid mites and termites preserved in Cretaceous amber reveals the evolutionary significance of phoresy Hemen Sendi, Pavel B. Klimov, Vasiliy B. Kolesnikov, Júlia Káčerová, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5389108/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Among minute-sized and wingless arthropods, astigmatid mites stand out for their diverse range of symbiotic associations (parasitic, neutral and mutualistic), with both invertebrate and vertebrate hosts. When inhabiting discontinuous and ephemeral environments, astigmatid mites adapt their life cycle to produce a phoretic heteromorphic nymph. When feeding resources are depleted, phoretic nymphs disperse to new habitats through phoresy, attaching to a larger animal which transports them to new locations. This dispersal strategy is crucial for accessing patchy resources, otherwise beyond the reach of these minute arthropods. In Astigmata, the phoretic nymph is highly specialized for dispersal, equipped with an attachment organ and lacking a mouth and pharynx. Despite the common occurrence of phoretic associations in modern mites, their evolutionary origins remain poorly understood. Among Astigmata, the family Schizoglyphidae represents an early derivative lineage with phoretic tritonymphs; however, our knowledge of this family is limited to a single observation. Results Here, we report the oldest biotic association of arthropods fossilised in amber (~130 Ma, Lebanon): an alate termite with 16 phoretic deutonymphs of Schizoglyphidae (Plesioglyphus lebanotermi gen. et sp. n.). The mites are primarily attached to the membranes of the host’s hindwings, using their attachment organs, pretarsal claws and tarsal setae. Additionally, we report new modern phoretic tritonymphs of this same family, on one of the earliest lineages of termites. These data collectively indicate that schizoglyphid-termite associations represent the oldest continuous mite-host associations. Notably, schizoglyphid mouthparts retain a distinct mouth and pharynx, absent in modern Astigmata. Conclusion The discovery of Schizoglyphidae mites in Lebanese amber represents the oldest known continuous association between acariform mites and their hosts. This finding demonstrates the long-term evolutionary significance of phoresy in Astigmata, evidencing a relationship sustained for over 130 Ma. It indicates that these early mites lived inside termite nests as inquilines and used alate termites for dispersal. This ancient association offers key insights into the coevolution of both mites and termites, highlighting a potential for the future discoveries of similar mites. This fossil —a stem-group Astigmata— is important for the accurate calibration of acariform mite phylogenies, advancing our understanding of these mites evolutionary history. Isoptera Lower Cretaceous Barremian social insects Lebanese amber Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Background Phoresy is a symbiotic interaction in which one life stage of a smaller animal attaches to a larger animal to facilitate dispersal [1-2] and access a more favourable habitat [3]. While reflecting primarily commensalism [4], phoresy can be defined together with other types of symbiotic interactions like mutualism [5], parasitism [6], or parastioidism [1, 7] as part of the lifestyle of the phoretic organism. For relatively small and/or wingless arthropods (springtails, pseudoscorpions and mites), phoresy is a key strategy to provide access to distant and patchy resources beyond their normal reach [8-9]. Phoretic organisms exhibit host-seeking behaviours and various adaptations for attachment; they typically do not feed, nor reproduce, during transport on their hosts [1]. Mites (Acari), with around 55,000 described species, are among the most diverse arthropods, representing a clade of substantial medical and economical importance. They exhibit a range of ecological lifestyles, including free-living, parasitism, active predation, and saprophagy particularly in soil-dwelling mites [10-12]. Modern mites, although they may not form a monophyletic group, include two monophyletic superorders: Acariformes (32,000 species) and Parasitiformes (23,000 species) [13]. Phoresy has evolved and disappeared multiple times throughout mite evolution [1]. Among the Acariformes, phoresy is common in Heterostigmata (2,700 species) and Astigmata (6,300 species) [1]. In Heterostigmata, females are typically the dispersal stage, whereas in most free-living Astigmata, the heteromorphic deutonymph—formerly known as the 'hypopus'—serves as the dispersal stage [14-15]. In the enigmatic astigmatid family Schizoglyphidae, the dispersal stage is likely a tritonymph [16], which morphologically resembles the heteromorphic deutonymph. Astigmatid heteromorphic nymphs possess a highly specialized attachment organ, featuring various suckers and adhesive conoids adapted to different attachment sites, such as smooth insect cuticle, setae, or mammal hair [15]. Astigmata have a shorter life cycle compared to their ancestors, oribatid mites. While oribatid mites inhabit soil—an uninterrupted habitat—Astigmata prefer discontinuous and ephemeral environments, such as decomposing plant and fungal materials, stored products, phytotelmata, dung, actively growing mycelia, subcortical spaces, tree sap flows, and invertebrate and vertebrate nests. This habitat preference makes long-distance dispersal a crucial component of their life cycle, and Astigmata estalish phoretic associations with larger organisms, such as mammals, insects, and myriapods [14]. Astigmatid phoretic nymphs often travel in groups to enhance sexual reproduction at their destination and increase their chances of establishing a large population on new resources, thereby outcompeting other colonizers [1]. Unlike other life stages (larvae, non-phoretic nymphs, and adults), phoretic nymphs are typically non-feeding, although there are occasional exceptions [6]. Adults are the sole reproductive stage. A few free-living Astigmata are capable of dispersing as adults [1, 14]. Phoresy is likely the ancestral lifestyle of Astigmata, but it has been lost in (i) astigmatids inhabiting continuous habitats (such as soil or water), (ii) associated with non-nest-building insects; or (iii) nearly all vertebrates with overlapping generations, which enables maternal vertical transmission [1, 14]. Astigmata have experienced three major evolutionary events: (1) the ancestral oribatid life cycle (lacking a phoretic deutonymph) evolved to a life cycle with a specialized phoretic deutonymph (non-schizoglyphid Astigmata) or tritonymph (schizoglyphid Astigmata); (2) the deutonymphal lifecycle was then modified to permanently suppress the deutonymphal stage in several lineages, notably in Psoroptidia, a lineage of full-time (permanent) associates of birds and mammals, as these mites could effectively colonize new hosts via vertical transmission and other direct host-to-host contacts; (3) Pyroglyphidae, a lineage within Psoroptidia, transitioned to living in the nests of their hosts, thereby becoming secondarily free-living and using their former hosts for transport without forming phoretic deutonymphs [1, 17-19]. The family Schizoglyphidae is the sister group to the remaining extant Astigmata [1]. This family is distinctive for retaining several plesiomorphic traits, including the relative position of the genital opening and attachment organ, as well as the structure of the attachment organ and gnathosoma compared to other astigmatid families [20]. This monotypic family includes a single observation of two specimens of Schizoglyphus biroi Mahunka, 1978, found in New Guinea (Indonesia) on a tenebrionid beetle. Although this species exhibits the specific morphological adaptations characteristic of a phoretic heteromorphic deutonymph, it is probably a tritonymph [16]. Evidence for this includes the presence of three pairs of genital papillae (as seen in oribatid tritonymphs and adults) instead of the two pairs found in deutonymphs and adults of other Astigmata. Despite substantial efforts to find this mite again on tenebrionid beetles, it has not been re-collected, suggesting that the original phoretic host (the beetle) may represent an incidental record rather than a true biologically relevant association. Thus, the true association of Schizoglyphus biroi and the family it represents remains elusive. Acariformes is one of the earliest diverging groups within the arachnids, with fossil evidence dating back to the early Devonian [21]. However, symbiotic associations with other organisms are mostly documented from the Cretaceous, when amber preserved these interactions in situ [14, 22-23]. Crown-group Astigmata are estimated to have diverged between the Late Permian and the Early Triassic, while the stem-group likely originated in the Late Devonian to Carboniferous [13-14, 17, 22-23]. In the fossil record, Astigmata appears from the Cretaceous onward, becoming more abundant in the Cenozoic, with several instances of biotic associations [24-32]. Here, we report the oldest amber association of an phoretic arthropod associated with its host preserved in Lebanese amber (Early Barremian, ~ 130 Ma): 16 mite deutonymphs belonging to the family Schizoglyphidae, phoretic on an alate termite Lebanotermes veltzi (Figs. 1; and 2). To confirm this association and explore potential true hosts of Schizoglyphus biroi , we also examined nests of modern termites in New Zealand and found schizoglyphids associated with Stolotermes , one of the earliest diverging termite lineages. The combined fossil and modern evidence suggests that schizoglyphid-termite associations represent the oldest known ongoing relationship between mites and their hosts. These findings shed light on the range of plesiomorphic features (including the gnathosoma and the attachment organ), present in the earliest crown-group Astigmata and reveal a highly conserved phoretic morphology that has persisted since the Early Cretaceous. This also advances our understanding of the temporal framework for the diverse mite-arthropod ecological associations observed today. Methods We studied 16 mite specimens attached to the fossil holotype of termite Lebanotermes veltzi described in Engel et al. [33]. The termite host which was preserved in a single amber piece (341 C–T) from the Lower Barremian of Mdeyrij-Hammana, Caza (District) Baabda, Lebanon, coll. D. Azar. The amber piece is housed at the Natural History Museum of the Lebanese University, Faculty of Sciences II, Fanar. Other syninclusions derived originally from the same amber block include: the allotype of the chironomid dipteran Ziadeus kamili (341 B), an aleyrodid hemipteran (341 A), and a ceratopogonid dipteran. The amber piece was trimmed and polished for microscopy, embedded in Canada balsam and placed in a cube of 1 mm thick microscopic glass slides. This permanent glass-amber preparation makes computed tomography not feasible for resolutions targeting the 150 µm-long mites. Several microphotographs (Figs. 1A, C; 2A–D, F; and 4A–C) were acquired using the protocol described on this website (https://enrico-bonino.eu): a Sony a7R II mirrorless camera with a 208 mm tube lens, a Raynox DCR-150, microscope objectives Mitutoyo QV 2.5× (or APO 20x), and a 110 mm tube lens with inversed Schneider Componon-S 50 mm/f2.8 lens for the whole specimen. This system (camera, tube-lens, and optics) is mounted on a MJKZZ Ultra Rail MINI V2, allowing movements in both vertical and horizontal planes. The illumination was provided by a cylindrical OGGLAB LED system DB 120EB. The entire panoramic image was assembled from two overlapping stacks, each composed of 53 frames. Images were captured in 16-bit RAW format with several steps between frames: 15 µm (with the Mitutoyo QV 2.5x), 2 µm (with the Mitutoyo 20x), and 95 µm (with Schneider-Componon lens). The frames were sub-stacked per group of eight images, stacked secondarily together and retouched in Helicon Focus (v.8.2, Professional) to remove undesired features (e.g. bubbles, dust, surfaces not in focus). Final image post-processing was done with Adobe Photoshop and Topaz DeNoise software. We also used a Zeiss Axiocam 208 mounted on an Axioimager A2 microscope, equipped with EC Epiplan 20x/0.4, 50x/0.55, and a W N-Achroplan 63x0.9 (water immersion) both in reflection and transmission light mode. Multispectral microimaging was performed at a research platform IPANEMA (SOLEIL Synchrotron, Orsay, France, microscope magnification x20). Reflection and luminescence images emitted by the sample were collected in various spectral ranges using a setting coupling (1) an illumination device employing 16 different LED lights (from 365 up to 700 nm wavelength, CoolLED pE-4000) and (2) a light filter device placed in front of the camera detector (a wheel holding six interference band-pass filters collecting signal within in six spectral ranges from 435 to 935 nm). Out of the 96 produced illumination/detection couples, a selection of three couples enhancing morphological features of interest were combined into pseudo-coloured RGB. The stacking, alignment, image registration of the different couples, and production of pseudo-coloured RGB composites were performed using ImageJ. Pseudo-coloured RGB images were produced with red—illumination 435 nm/detection 650 ± 60 nm (luminescence), green—470 nm/det. 650 ± 60 nm, blue—470 nm/det. 732 ± 68 nm. Results 1. Systematic paleontology Class Arachnida Cuvier, 1812 Superorder Acariformes Zachvatkin, 1952 Order Sarcoptiformes Reuter, 1909 Suborder Oribatida Dugès, 1834 Hyporder Astigmata Canestrini, 1891 Family Schizoglyphidae Mahunka, 1978 (type genus Schizoglyphus Mahunka, 1978) Plesioglyphus gen. n. ZOOBANK CODE Type species. Plesioglyphus lebanotermi gen. et sp. n. Type material. Holotype (341E): phoretic tritonymph, specimen 1 (Figs. 2A; 3; and 5C), attached to alate specimen of the termite Lebanotermes veltzae Engel, Azar et Nel, 2011 (holotype), embedded in single amber piece (341 C–T), the Lower Barremian of Mdeyrij-Hammana, Caza Baabda, Lebanon, coll. D. Azar, preserved at the Natural History Museum of the Lebanese University, Beirut. Paratypes: 15 phoretic deutonymphs (341 F–T), same data. Type locality and horizon. Mdeyrij-Hammana, Caza (District) Baabda, Mount Lebanon Governorate, Lebanon, Lower Cretaceous, Lower Barremian. Diagnosis. Tritonymph . Subcapitular remnant large, with 2 pairs of short adoral setae (Fig. 3D). Palps free, long, 2-segmented; palp tarsus with at least 1 solenidion (Figs. 4F; and 5). Dorsal idiosoma sclerotized, punctate; sejugal furrow well developed. Progenital and anal opening well separated. Coxal fields III medially separated, distance between them is distinctly longer than the width of the trochanter III (Fig. 5B–D). Anterior apodemes IV curved in the medial part, distinctly angular. Attachment organ large, with anterior suckers ( ad 3 ) and median suckers ( ad 1+2 ) well developed (Figs. 3B; .4D–F; and 5B,–D); conoids ps 1 and ps 2 vestigial. Legs with typical segmentation (trochanter-tarsus). Tarsal empodial claws I-IV present, arising directly from tarsal apices (Fig. 5). Some tarsal setae foliate (Figs. 4F; and 5). Remarks . Plesioglyphus belongs to Schizoglyphidae based on the presence of 2 pairs of adoral setae, well-developed palps, 4 pairs of genital setae, transversely elongated cuticular suckers formed by the fusion of pseudanal setae p 1 +p 2 , and the anal opening situated between suckers ad 1+2 . Plesioglyphus gen. n. is very similar to the extant genus Schizoglyphus , but differs by the following: the gnathosoma is larger, reaching femora I (distinctly not reaching in Schizoglyphus ); the distance between suckers ad 3 and ad 1+2 is slightly smaller than the diameter of these suckers (slightly larger than the diameter of ad 1 in Schizoglyphus ); anterior apodemes IV are distinctly curved medially (curved only at tips in Schizoglyphus ). Etymology. The generic name is formed from two Greek stems, πλησίον (near, neighbouring) and γλῠ́φω (to carve, cut out with a knife, engrave). The former stem is used in the formation of names in paleontology, while the latter stem is widely used to form names in astigmatid mites. Gender masculine. Plesioglyphus lebanotermi sp. n. ZOOBANK CODE Description. Tritonymph . Gnathosoma large, subcapitular remnant subquadrate (width 1.2 times longer than length) (Fig. 3D). Palps 2-segmented. Palptarsus with at least one a seta and a solenidion ω; solenidion ω subequal or longer than palps (Figs. 3B–C; 4F; and 5). Two pairs short adoral setae present (Fig. 3D). Gnathosoma (except for palps) situated under rostrum. Rostrum large, wide, rounded, without eyes. Dorsum with propodosomal and hysterosomal shields. Shields roughly punctate, separated by a distinct sejugal furrow. Dorsal setae not observed, except h 3 on posterior hysterosoma. Ventral side. Sternum present, long, nearly reaching anterior portions of apodemes II (specimen 12). Coxal apodemes II-IV with free ends, anterior apodemes IV distinctly curved medially, angular. Coxal fields I-IV open; coxal fields III well-separated medially, distance between them distinctly longer than width of trochanter III. Coxal setae not observed. Progenital opening nearly as long as base of legs III or IV, situated at level of trochanters IV, well-separated from attachment organ. Genital setae present, at least 4 pairs (Fig. 3B). Genital papillae not observed. Anal opening situated within attachment organ; distance between anal and progenital openings more than twice longer the length of progenital opening. Attachment organ large, almost as wide as body width. Suckers ad 3 and ad l+2 large, oval; ad 3 posterior to progenital opening, ad l+2 lateral to anal opening (Figs. 3B; 4D–F; and 5B–D). Distance between ad 3 and ad l+2 shorter than diameter of these suckers. Dorsoventral muscles of suckers ad l+2 well-developed (holotype). Alveolae ps 1 and ps 2 situated between suckers ad 3 and ad l+2 . Legs short, thick, with typical set of segments. Empodial claws I-IV present, slightly shorter to tarsi (Fig. 5A,B). Tarsus I with seta e , long, widened at tip and at least 1-2 other foliate setae, other setae spiniform. Tarsus II similar to tarsus I, but e shorter. Tarsi III and IV with at least 6 foliate setae; 1 seta on tarsus IV longer than other (Figs. 4F; and 5C–E). Tibiae I-II with 2 setae ( gT and hT ) (Fig. 5,E), setae on tibiae III-IV not observed. Femora I-II with seta vF (Fig. 5C). Setation of tibiae III-IV, genua I-IV, femora III-IV and trochanters I-IV not observed. Tarsi I-II with 2 solenidia ω. Tibiae I-II with 1 long solenidion φ (its tip reaches tip of seta e ) (Figs. 4F; and 5). Tibia III with a single solenidion φ (longer than combined length of genu and tarsus III) (Figs.3C; and 5). Genu I with one bacilliform solenidion σ (Fig. 5E). Other solenidia not observed. Measurements (n=3). Idiosoma 182–220 long, 105–110 wide. Prodorsum 68, width 95. Hysterosoma length 115. Gnathosoma 30–35, width 23–25; free palps 14–15, gnathosomal solenidion ω 14–18. Length of attachment organ 50–67, width 66–70, ad 3 17–22 x 13–19, ad l+2 20–28 x 17–24. Legs I: length 46–50, e I 24–30, ω I 9–11, φ I 34–36, σ I 10. Legs II: length 37, e II 14–22, φ II 20. Remarks. The new species differs from Schizoglyphus biroi by long tibial solenidia φ I–II protruding the tips of respective tarsi (not protruding in S. biroi ). The following 16 phoretic deutonymphs were examined: Specimen 1 (341 E) (Fig. 2A; 3; and 5C) – holotype, ventral. Legs and most of tarsal setae, gnathosoma (including adoral setae) and muscles are visible; a bubble inside the mite obscures observation. Specimen 2 (341 F) (Figs 2A; 4E–F; and 5E) – paratype, dorsolateral. Patterns on the dorsal shields and the posterior part of the attachment organs are well visible; the palp tarsus; legs I, right legs II -IV, anal opening, suckers ad 3 and ad l+2 are somewhat visible. Specimen 3 (341 G) (Fig. 2B) – paratype, frontal view. Outlines of the gnathosoma, legs I, and sejugal furrow were observed. Specimen 4 (341 H) (Figs 2C; 4D; and 5D) – paratype, ventral. The gnathosoma, suckers ad 3 and ad l+2 , ps 1 , anal and genital opening, some coxal apodema, and right tibial solenidion φ II are visible; legs are somewhat visible as outlines. Specimens 5–11 (341 G–O), 13 (341 Q), 14 (341 R) (Fig. 2 E–F) – paratypes, ventral (5, 8), dorsolateral (6), lateral (7, 9, 13, 14), dorsal (10, 11). In specimens 10 and 11, hysterosomal and propodosomal shields (with rostrum), legs I-II, and sejugal furrow are somewhat visible. Other specimens are poorly visible. Specimen 12 (341 P)(Fig. 4C) – paratype, ventral. Gnathosoma, legs I-II, genital and anal openings, apodemes I-II, attachment organ, legs III-IV are somewhat visible. Specimen 15 (341 R)(Fig. 4B) – paratype, ventral. The progenital and anal openings, suckers ad l+2 and left ad 3 , coxal areas are well visible. Legs I-III are somewhat visible. Specimen 16 (341 T) (Figs 2F; 4A; and 5A) – paratype, dorsal. The dorsal shields, their sculpture, sejugal furrow, legs I-II with empodial claws are well visible. One dorsal seta (probably h 3 ) is somewhat visible. 2. Evidence for a true phoretic association preserved in amber The termite Lebanotermes veltzae was preserved carrying 16 mites, positioned both dorsally (Fig. 1A–B) and ventrally (Fig. 1C). Specimens 1–4 are located at various points on or near the termite's body, while specimens 5–16 form a larger cluster over its abdominal wing area (Fig. 1B). Although some specimens were dislodged from the host (specimens 1-3, possibly during the process of amber entrapment, other specimens still remain attached to the wing membrane (specimens 4-16). Notably, specimen 16 has its pretarsal claws, equipped with foliate setae, in direct contact with the membrane of the right hindwing's dorsal side (Figs. 2D, 2F; and 4A). The presence of 16 conspecific phoretic deutonymphs on the termite host, along with evidence of direct mite-host contact, supports the conclusion that this association is neither taphonomic nor random, but represents a true biological relationship. 3. Modern schizoglyphids are found in termite associations The earliest known crown-group astigmatid, Schizoglyphus biroi , a heteromorphic phoretic tritonymph, was found on the tenebrionid beetle Dioedus tibialis (= Tagalus tibialis ) in western New Guinea [35]. Despite extensive efforts to re-collect these mites from similar hosts (OConnor, pers. comm.), no further specimens were found, suggesting that this record was incidental rather than indicative of a true biological association. Another reported occurrence of " Schizoglyphus sp." on a scarabaeid larva from India [36] actually represents the genus Sancassania (family Acaridae). Here, we report the discovery of five deutonymphs of Schizoglyphus sp. found in 2022 galleries of the New Zealand wetwood termite Stolotermes ruficeps (family Stolotermitidae), in a Pinus radiata log in Thames, New Zealand (Dickson Holiday Park, Tinker Trail, 37°06'42.7"S 175°31'22.1"E). The termite specimens were ethanol-washed from alate females, workers, and immature termites. These mites display several key character states that unequivocally place them within Schizoglyphidae: three-segmented palps (Fig. 6C), three pairs of genital papillae, five pairs of genital setae (Fig. 6B and D), the gnathosoma bearing adoral setae (Fig. 6B–C), the cuticular suckers of the attachment organ are composed of p 1 +p 2 (with alveoli), and the anal opening situated between suckers ad 1+2 (Fig. 6B and D). These specimens represent a new species, which will be described in a future publication. Additionally, a different species of deutonymphal Schizoglyphus sp. was previously collected from an alate queen of Stolotermes ruficeps in New Zealand, although these specimens were unfortunately lost (B. OConnor, pers. comm.). These new findings strongly suggest that schizoglyphids live inside termite nests, likely as inquilines, and use founder alate termites for dispersal from one nest to another. Discussion The nests of social insects, such as termites, are stable, long-term habitats that provide abundant food resources, attracting a diverse array of inquiline organisms, including mites, flies, beetles, antlions, wasps, true bugs, silverfish, springtails, woodlice, harvestmen, pseudoscorpions, spiders, millipedes, and gastropods [9, 37-39]. Among these, Astigmata stands out as the most speciose lineage of termitophiles (see also Table 1 in supplementary materials) [40]. Remarkably, very few astigmatans were reported on modern alate termites, compared to fossil record. We are confident interpreting it as a pure observation bias. First, because there are limitations in noticing them easily on alate hosts. Indeed, when mites are phoretic of alate reproducers of social insects (like the astigmatan Lemanniella sp. on the ant Messor pergandei ), the phoront often is hidden underneath the wings, at body contact (as in the bee Halictus frontalis ) [41]. Second, because amber has already proven to preserve phoretic associations which went nowadays discovered only after a fossil occurence had suggested their existence. That is the case of some larger wingless microarthropods (springtails), that were reported phoretic on diverse insect hosts in amber, including on alate social insects with the prediction that these interactions shall still exist [e.g. 38, 42-43]. Modern springtails were since indeed observed from the body of flies, tipulids and alate termites (N.R. pers. obs.). Indeed, alates (reproducers) show a quite short lifespan compared to other casts of social insects: produced only at a certain time of year, and losing their wings right after nuptial flight, when flying away to form a new colony. In contrast, alate casts in amber are not uncommon (both males and females), with resin patches forming along the barks mostly above ground level, trapping the flying individuals. This is illustrated by amber termite-mite records: unidentified mites and Mesostigmata were respectively found on extinct Euisoptera [38] from Jordanian amber with a similar Early Barremian age as Lebanese amber [44] (~ 130 Ma) and a Cenomanian (~ 100Ma) termite-like roach from Burmese amber [45]. Both records indeed involve alate adults as well (female in post-nuptial flight in the second case). Astigmatan heteromorphic nymphs, including Plesioglyphus lebanotermi , have a highly specialized attachment organ (Figs. 4–6) serving for attachment and hitchhiking on hosts [14] [46]. The success of a deutonymph attachment depends on its location on the host's body, to both prevent detachment by the host and minimize interference with the host's locomotion abilities [47]. Among the 16 individuals of P. lebanotermi , most specimens were found on wings, on areas of overlap between hindwings (sp. 5–15, Fig. 1; and 2D), or between hindwing and forewings (sp. 16, Figs. 1; 2F; and 5E) suggesting that the fossil deutonymphs were in this position during their transport. As astigmatid heteromorphic deutonymphs lack a mouth, oral feeding is not possible. Still feeding as a parasite can occur during phoresy via attachment organ suckers, anus or genital papillae [1,48]. Inside the nest, certain mites (such as Australhypopus sp.) may provide sanitary functions by feeding on dead termite corpses [5-6]. During phoresy, mites can potentially impede the mobility of their hosts when the mite loads are high [49-50]; but, in general, phoresy is harmless to them. The significant advantage gained by phoronts is linked to an increased dispersal distance, allowing them to exploit new food resources or access different hosts. In social insects, dispersal is enhanced during periods of colony swarming through attachment to alate reproducers. At swarming, alate termites can travel distances of over one kilometre [51]. The observed ~ 130 Ma old association of Plesioglyphus lebanotermi with Lebanotermes veltzae is here most likely to represent phoretic commensalism, while non-phoretic stages of the mites likely live inside termite nests as commensals. Four distinct superfamilies of Astigmata are associated with four different termite lineages (Fig. 7; Table 1 in supplementary materials). Among these, the superfamily Acaroidea is the most commonly reported, with 13 genera and 21 species identified from members of the Rhinotermitidae (including Psammotermes hypostoma , Reticulitermes flavipes , and Coptotermes formosanus ) and one Termitidae species ( Cornitermes cumulans ) (Fig. 7; Table 1 in supplementary materials). The superfamily Histiostomatoidea has been recorded on several rhinotermitid species, while a species of the superfamily Hemisarcoptoidea has been found on Psammotermes hypostoma (Rhinotermitidae). The frequent association of Acaroidea with termites is expected, as this superfamily is species-rich, comprising 562 species [1]. However, despite comparable species diversity (576 species), there are fewer reports of phoretic associations involving Histiostomatidae. Hemisarcoptoidea is a smaller group, with 144 species. All three of these mite superfamilies are associated with Neoisopteran termites, which are believed to have diverged approximately 110 million years ago (with the split between Termopsidae and Rhinotermitidae occurring around 85 Ma [51], Fig. 7). In contrast, both modern and fossil schizoglyphoid associations, including those reported here, are found on termite lineages that diverged much earlier—from the Early Barremian (126 Ma for Stolotermitidae) to the Late Lias (185 Ma for Lebanotermes, Euisoptera; [52]). This distribution may suggest a degree of specialization among phoretic astigmatid mites for specific termite lineages, possibly selected through co-occurrence over geological time and maintained to the present day (Fig. 7). However, the limited number of schizoglyphid occurrences currently prevents any definitive assessment of this pattern. Schizoglyphid phoretic nymphs retain several plesiomorphic traits when compared to other astigmatid mites. These include: (i) A gnathosomatic remnant with two pairs of adoral setae and relatively long palps, whereas other astigmatids lack adoral setae and have reduced palps; (ii) Three pairs of genital papillae (though this was not observed in Plesioglyphus lebanotermes ) and four or more pairs of genital setae, while other astigmatids typically have two pairs of genital papillae and only one pair of genital setae; (iii) Pseudanal setae p 1 +p 2 are fused into large, transversely elongated cuticular suckers, unlike the small, rounded suckers seen in most astigmatids; (iv) The anal opening is positioned more posteriorly than in most astigmatids, situated at the level of ad 1+2 , rather than ad 3 as is typical in other astigmatids. There are interesting similarities between Plesioglyphus the phoretic nymphs of the genus Levantoglyphus (family Levantoglyphidae), recently reported from Lebanese amber without host information [13]. Both genera share a well-developed gnathosomal remnant with long palps and long terminal solenidia (ω), indicating the importance of host-seeking behaviour in both lineages. However, Levantoglyphus possesses rudimentary chelicerae, enabling food shredding in non-phoretic stages, whereas modern schizoglyphids from New Zealand retain a mouth and pharynx, suggesting that the reduction of functional mouthparts in astigmatid phoretic nymphs was a gradual evolutionary process. Plesioglyphus lebanotermi displays all the synapomorphies of modern schizoglyphid mites and documents the existence of non-feeding, phoretic heteromorphic nymphs in astigmatid mites from the Early Cretaceous (~130 Ma). This fossil represents the earliest known crown-group Astigmata with a confirmed phoretic association with termites—a relationship that has persisted into modern times. Its placement among living mites will allow for precise calibration of molecular clock phylogenies. In contrast, the transitional deutonymphs of the extinct stem-group family Levantoglyphidae , whose host associations remain unknown, provide a less precise calibration for molecular clock phylogenies. As these deutonymphs belong to a stem-group lineage of Astigmata, their use in calibrating molecular clocks is limited to a broader and less specific range compared to Plesioglyphus lebanotermi . Conclusion The oldest known biotic association of arthropods preserved in amber, dating to approximately 130 million years ago, involves Astigmata, a group of mites specialized in phoresy, which were found attached to a winged termite. This discovery represents the earliest known instance of phoretic mites associated with an arthropod. The mites belong to the genus Plesioglyphus n. gen. of the family Schizoglyphidae, an early-diverging lineage of Astigmata currently recognized from a single described species. Remarkably, the plesiomorphic features of these ancient mites —such as long palps and a large subcapitulum— have been highly conserved over 130 million years. In the Early Cretaceous of Lebanon, these schizoglyphids coexisted with other extinct Astigmata ( Levantoglyph us) that also exhibited plesiomorphic mouthparts. However, unlike these extinct relatives, the schizoglyphids described here are the earliest known crown-group Astigmata to display strictly non-feeding, heteromorphic nymphs. This finding reveals a specialized phoretic relationship with their termite host, suggesting that Plesioglyphus functioned as an inquiline in its feeding stages, and dispersed on winged termites via phoresy during non-feeding, heteromorphic stages. This association, which has persisted into modern times, highlights the long-standing evolutionary relationship of Schizoglyphidae with eusocial insects — a connection that has been largely overlooked. Our findings show the remarkable diversity and evolutionary persistence of modern Schizoglyphidae, which continue to exhibit phoresy on termites, reflecting their ancient and ongoing relationship with these insects. Declarations Ethics approval and consent to participate Field studies were conducted in accordance with local legislation. As for Lebanon, the amber collecting was performed after obtention by Pr. Dany Azar of the necessary authorisations from the Municipality of Hammana, the Lebanese Ministry of Power, Mining direction; and the National Council for Scientific Research - Lebanon (National central public institution in charge of science policy-making under the authority of the President of the Council of Ministers). Consent for publication All the reported information was used with the consent of their owners. Availability of data and materials All data supporting these findings are included in this article. Competing interest The authors declare no competing interests. Funding This research was supported by a BELSPO BRAIN-be federal Belgian grant (B2/202/P1/PARADI2S) and by the EU NextGenerationEU through the Recovery and Resilience Plan for Slovakia under the project No. 09I03-03-V04-00439. P.B.K. and V.B.K. were supported by the Ministry of Science and Higher Education of the Russian Federation within the framework of the Federal Scientific and Technical Program for the Development of Genetic Technologies for 2019-2027 (agreement No. 075-15-2021-1345, unique identifier RF----193021X0012). This paper is a contribution to the activity of the laboratory ‘Advanced Micropalaeontology, Biodiversity and Evolution Researches (AMBER)’ led by DA at the Lebanese University. Authors’ contributions Conceptualization and methodology by N.R., H.S., P.K.. Formal analysis and visualization by H.S., N.R., V.K., P.K., E.B. and J.K.. Investigation and writing of the original draft by H.S., N. Robin, P.K. and E.B.. Writing - Review & Editing by V.K. and D.A.. Resources by D.A. and P.K.. Project administration and supervision by N. R. and D.A. All authors confirm their authorship and approve the final version of the manuscript. Acknowledgements We thank Dmitry Vorontsov for technical help, Barry OConnor for his record of the additional modern deutonymphal Schizoglyphus , Michael Engel for valuable scientific advice on termites, and Peter Vršanský for providing amber samples with evidence of mite syninclusions along with cockroaches. References 1. Seeman OD, Walter DE. Phoresy and mites: More than just a free ride. Annu Rev Entomol. 2023;68:69–88. https://doi.org/10.1146/annurev-ento-120220-013329 2. Camerik AM. Phoresy revisited. In: Sabelis MW, Bruin J, editors. Trends in Acarology. Dordrecht: Springer Netherlands; 2010. p. 333–6. 3. White PS, Morran L, de Roode J. Phoresy. Curr Biol. 2017;27:578–80. https://doi.org/10.1016/j.cub.2017.03.073 4. Myles TG. 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Mites are positioned as follows: 1, 4, 5, 8, 12, 15 ventral; 10, 11, 16 dorsal; 2, 6, 7, 9, 13, 14 dorsolateral; 3 frontal. \u003cstrong\u003e(C)\u003c/strong\u003eVentral view of termite with highlighted occurrence of a mite on the host foreleg. Boxes refer to detailed photographs in Fig. 2. Scale bars: 5 mm.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5389108/v1/0fcb8c635fe0a51ad2e0f803.png"},{"id":69463675,"identity":"ae1a35a6-7961-474e-a16d-e71bfe72d665","added_by":"auto","created_at":"2024-11-20 15:14:06","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1063435,"visible":true,"origin":"","legend":"\u003cp\u003eAttachment of the schizoglyphid mite \u003cem\u003ePlesioglyphus lebanotermi\u003c/em\u003e \u003cstrong\u003egen. et sp. n.\u003c/strong\u003e on the termite from Lebanese amber, Barremian. \u003cstrong\u003e(A)\u003c/strong\u003e Specimens 1 and 2 detached ventrolaterally from the termite host. \u003cstrong\u003e(B)\u003c/strong\u003e Specimen 3 is attached to the forefemur of the termite host. \u003cstrong\u003e(C)\u003c/strong\u003e Specimen 4 is attached between the hindwings of the host. \u003cstrong\u003e(D)\u003c/strong\u003eSpecimens 9-15 are attached between the termite hindwings above the dorsal abdominal area. \u003cstrong\u003e(E)\u003c/strong\u003e Specimens 5-8 are attached between the termite hindwings above the dorsal abdominal area. \u003cstrong\u003e(F)\u003c/strong\u003e Detail of attachment of specimen 16 on the wing membrane of its host. \u003cstrong\u003e(G)\u003c/strong\u003e– +/- schematics of the CuA area of an extant termite hindwing adapted from Scheffrahn \u003cem\u003eet al.\u003c/em\u003e [34] (CC BY 4.0) similar to the area with attached mites in \u003cem\u003eLebanotermes veltzae\u003c/em\u003eEngel, Azar et Nel, 2011. Note that the wings of termites are not as corrugated as in Paleozoic insects or in Odonatoptera, and Blattodea. Scale bars: 500 μm (A, D), 100 μm (B–C), 0,3 mm (E), 150 μm (F), 2 mm (G).\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5389108/v1/bcc1d4a681212079bf4762b9.png"},{"id":69463676,"identity":"16b06614-ff8f-4bec-b4c4-392451ecc247","added_by":"auto","created_at":"2024-11-20 15:14:06","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":847316,"visible":true,"origin":"","legend":"\u003cp\u003eLight microscope images of the schizoglyphid deutonymph of\u003cem\u003e Plesioglyphus lebanotermi\u003c/em\u003e \u003cstrong\u003egen. et sp. n\u003c/strong\u003e., holotype (341C), Lower Barremian Lebanese amber of Mdeyrij-Hammana, ventral view \u003cstrong\u003e(A)\u003c/strong\u003eTotal view. \u003cstrong\u003e(B)\u003c/strong\u003e Posterior part. \u003cstrong\u003e(C)\u003c/strong\u003e Left legs III and IV. \u003cstrong\u003e(D)\u003c/strong\u003e Anterior part and gnathosoma. Abbreviations: \u003cem\u003ead – \u003c/em\u003esuckers; φ \u003cem\u003e–\u003c/em\u003e tibial solenidion; \u003cem\u003eg – \u003c/em\u003egenital setae; \u003cem\u003eor –\u003c/em\u003eadoral setae. Scale bars 20 μm.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5389108/v1/9219df2903dcdf2c00846ca1.png"},{"id":69463677,"identity":"a827ded4-0acc-48bd-8fff-b2c05e6c8984","added_by":"auto","created_at":"2024-11-20 15:14:06","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":843628,"visible":true,"origin":"","legend":"\u003cp\u003eLight microscope images of the schizoglyphid deutonymph\u003cem\u003e Plesioglyphus lebanotermi\u003c/em\u003e \u003cstrong\u003egen. et sp. n\u003c/strong\u003e. from Lower Barremian Lebanese amber of Mdeyrij-Hammana \u003cstrong\u003e(A)\u003c/strong\u003e Dorsal view of specimen 16. \u003cstrong\u003e(B)\u003c/strong\u003e Ventral view of specimen 15. \u003cstrong\u003e(C)\u003c/strong\u003e Ventral view of specimen 12, paratype. \u003cstrong\u003e(D)\u003c/strong\u003e Ventral view of specimen 4. \u003cstrong\u003e(E-F)\u003c/strong\u003e Dorso-lateral view of specimen 2. Abbreviations: \u003cem\u003ead – \u003c/em\u003esuckers; ω \u003cem\u003e–\u003c/em\u003e palp solenidion; φ \u003cem\u003e–\u003c/em\u003e tibial solenidion; \u003cem\u003ee\u003c/em\u003e \u003cem\u003e– \u003c/em\u003etarsal I seta. Scale bars 20 μm (A, D–F), 120 μm (B), 60 μm (C).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5389108/v1/89f2d4ab89f1df0afc74795a.png"},{"id":69463673,"identity":"61ed1cc4-b96c-46ff-8a54-c8a81ae3d54f","added_by":"auto","created_at":"2024-11-20 15:14:06","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":411556,"visible":true,"origin":"","legend":"\u003cp\u003eLine drawings of the schizoglyphid deutonymph\u003cem\u003e Plesioglyphus lebanotermi\u003c/em\u003e \u003cstrong\u003egen. et sp. n\u003c/strong\u003e. from Lower Barremian Lebanese amber of Mdeyrij-Hammana \u003cstrong\u003e(A)\u003c/strong\u003e Dorsal view of specimen 16. \u003cstrong\u003e(B)\u003c/strong\u003e Ventral side of specimen 15. \u003cstrong\u003e(C)\u003c/strong\u003e Ventral side of specimen 1, holotype. \u003cstrong\u003e(D)\u003c/strong\u003eVentral side of specimen 4. \u003cstrong\u003e(E)\u003c/strong\u003eDorso-lateral side of specimen specimen 2. \u003cstrong\u003e(F)\u003c/strong\u003eColor codes for C-E: 1 – mite, 2 – presumably mite, 3 – mite internal structure, 4 – unknown structures, 5 – artifacts and sub-inclusions, 6 – outer borders of shadows. Abbreviations: Arrowheads indicate empodial claws. Abbreviations: \u003cem\u003ead – \u003c/em\u003esuckers; ω \u003cem\u003e–\u003c/em\u003e palp and tarsal solenidia; φ \u003cem\u003e–\u003c/em\u003e tibial solenidion; \u003cem\u003ee\u003c/em\u003e \u003cem\u003e– \u003c/em\u003etarsal apical seta\u003cem\u003e; cox – \u003c/em\u003ecoxa; \u003cem\u003eh – \u003c/em\u003edorsal setae; \u003cem\u003egT\u003c/em\u003e \u003cem\u003e– \u003c/em\u003etibial seta; \u003cem\u003ehT\u003c/em\u003e \u003cem\u003e– \u003c/em\u003e\u0026nbsp;tibial seta; \u003cem\u003evF – \u003c/em\u003efemoral setae\u003cem\u003e; ps\u003c/em\u003e \u003cem\u003e– \u003c/em\u003epseudanal setae;\u003cem\u003e g – \u003c/em\u003egenital setae; σ\u003cem\u003e – \u003c/em\u003e\u0026nbsp;genual solenidion. Scale bars: 100 μm.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-5389108/v1/d4312ae2aebbcd968bf0bc32.png"},{"id":69463678,"identity":"66667849-602a-4763-aacf-ad96a4e78c69","added_by":"auto","created_at":"2024-11-20 15:14:06","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1002683,"visible":true,"origin":"","legend":"\u003cp\u003eDiagnostic characters of phoretic nymphs of the family Schizoglyphidae as exemplified by a modern species collected from the termite \u003cem\u003eStolotermes ruficeps \u003c/em\u003efrom\u003cem\u003e \u003c/em\u003eNew Zealand. \u003cstrong\u003e(A)\u003c/strong\u003e Dorsal view. \u003cstrong\u003e(B)\u003c/strong\u003e Ventral view. \u003cstrong\u003e(C)\u003c/strong\u003e Ventral view of gnathosoma. \u003cstrong\u003e(D)\u003c/strong\u003e Attachment organ. Abbreviations: \u003cem\u003evi\u003c/em\u003e, \u003cem\u003eve\u003c/em\u003e, \u003cem\u003esi\u003c/em\u003e, \u003cem\u003ese\u003c/em\u003e– prodorsal setae; \u003cem\u003escx\u003c/em\u003e – supracoxal seta; \u003cem\u003ec\u003c/em\u003e, \u003cem\u003ed\u003c/em\u003e, \u003cem\u003ee\u003c/em\u003e, \u003cem\u003ef\u003c/em\u003e– hysterosomal setae; \u003cem\u003e1a\u003c/em\u003e, \u003cem\u003e3a\u003c/em\u003e, \u003cem\u003e4a\u003c/em\u003e, \u003cem\u003e4b\u003c/em\u003e – coxal setae; \u003cem\u003eg\u003c/em\u003e– genital setae; \u003cem\u003ead – \u003c/em\u003esuckers; \u003cem\u003eps\u003c/em\u003e \u003cem\u003e– \u003c/em\u003epseudanal setae; \u003cem\u003eor\u003c/em\u003e – adoral setae; \u003cem\u003eh\u003c/em\u003e – hysterosomal and gnathosomal setae; ω \u003cem\u003e–\u003c/em\u003e palpal solenidion; \u003cem\u003esup\u003c/em\u003e, \u003cem\u003ecm\u003c/em\u003e, \u003cem\u003eul\u003c/em\u003e, \u003cem\u003esul\u003c/em\u003e – palpal setae. Scale bar 100 µm (A, B), 20 µm (C), 50 µm (D).\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-5389108/v1/87391086aeef9034e9e377da.png"},{"id":69463674,"identity":"16e527de-03c5-4c9f-946d-44efc1908d9d","added_by":"auto","created_at":"2024-11-20 15:14:06","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":729957,"visible":true,"origin":"","legend":"\u003cp\u003eSimplified phylogenies of Astigmata and Isoptera correlated on a geological time scale with marked termite-mite associations and a reconstruction of the described termite-mite association. For a full list of Astigmata/termite associations see Table 1 in supplementary materials. Red dots indicate the herein described mite/termite association and grey boxes the other (extant) associations. Each association is indicated by a number. Grey scale bars reveal occurrence of the clade based on molecular data, while red lines indicate fossil calibrations. Note that the phylogeny of Astigmata is morphologically based as superfamilies appear to be non-monophyletic. Therefore, their origin based on molecular data might differ somewhat in reality. Data for Astigmata phylogeny is used from Seeman \u0026amp; Walter [5] and for Isoptera phylogeny from Jouault \u003cem\u003eet al.\u003c/em\u003e [52]. The reconstruction was made by Júlia Káčerová.\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-5389108/v1/62c4ed1766110934512d87ee.png"},{"id":69463932,"identity":"02f6da66-4ded-4dd6-ad3d-b13b4c52cbb5","added_by":"auto","created_at":"2024-11-20 15:22:14","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":7472576,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5389108/v1/12c5e4a5-6ce7-46f9-b958-5d9f0547cf9e.pdf"},{"id":69463931,"identity":"67c787c0-9e20-4401-a9c3-1bd20a4041c7","added_by":"auto","created_at":"2024-11-20 15:22:06","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":42099,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Materials\u003c/p\u003e","description":"","filename":"SILongtermphoresyinAstigmata9.2.docx","url":"https://assets-eu.researchsquare.com/files/rs-5389108/v1/780af451e6fdd30757546296.docx"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eThe oldest continuous association between astigmatid mites and termites preserved in Cretaceous amber reveals the evolutionary significance of phoresy\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"Background","content":"\u003cp\u003ePhoresy is a symbiotic interaction in which one life stage of a smaller animal attaches to a larger animal to facilitate dispersal [1-2] and access a more favourable habitat [3].\u0026nbsp;While reflecting\u0026nbsp;primarily commensalism [4], phoresy can be defined together with other types of symbiotic interactions like mutualism [5],\u0026nbsp;parasitism [6], or parastioidism [1, 7] as part of the lifestyle of the phoretic organism.\u0026nbsp;For relatively small\u0026nbsp;and/or wingless arthropods (springtails, pseudoscorpions and mites), phoresy is a key strategy to provide access to distant and patchy resources beyond their normal reach [8-9]. Phoretic organisms exhibit host-seeking behaviours and various adaptations for attachment; they typically do not feed, nor reproduce, during transport on their hosts [1].\u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Mites (Acari), with around 55,000 described species, are among the most diverse arthropods, representing a clade of substantial medical and economical importance. They exhibit a range of ecological lifestyles, including free-living, parasitism, active predation, and saprophagy particularly in soil-dwelling mites [10-12]. Modern mites, although they may not form a monophyletic group, include two monophyletic superorders: Acariformes (32,000 species) and Parasitiformes (23,000 species) [13]. Phoresy has evolved and disappeared multiple times throughout mite evolution [1]. Among the Acariformes, phoresy is common in Heterostigmata (2,700 species) and Astigmata (6,300 species) [1]. In Heterostigmata, females are typically the dispersal stage, whereas in most free-living Astigmata, the heteromorphic deutonymph\u0026mdash;formerly known as the \u0026apos;hypopus\u0026apos;\u0026mdash;serves as the dispersal stage [14-15]. In the enigmatic astigmatid family Schizoglyphidae, the dispersal stage is likely a tritonymph [16], which morphologically resembles the heteromorphic deutonymph. Astigmatid heteromorphic nymphs possess a highly specialized attachment organ, featuring various suckers and adhesive conoids adapted to different attachment sites, such as smooth insect cuticle, setae, or mammal hair [15].\u0026nbsp;Astigmata have a shorter life cycle compared to their ancestors, oribatid mites. While oribatid mites inhabit soil\u0026mdash;an uninterrupted habitat\u0026mdash;Astigmata prefer discontinuous and ephemeral environments, such as decomposing plant and fungal materials, stored products, phytotelmata, dung, actively growing mycelia, subcortical spaces, tree sap flows, and invertebrate and vertebrate nests. This habitat preference makes long-distance dispersal a crucial component of their life cycle, and Astigmata estalish phoretic associations with larger organisms, such as mammals, insects, and myriapods [14]. Astigmatid phoretic nymphs often travel in groups to enhance sexual reproduction at their destination and increase their chances of establishing a large population on new resources, thereby outcompeting other colonizers [1]. Unlike other life stages (larvae, non-phoretic nymphs, and adults), phoretic nymphs are typically non-feeding, although there are occasional exceptions [6]. Adults are the sole reproductive stage. A few free-living Astigmata are capable of dispersing as adults [1, 14].\u003c/p\u003e\n\u003cp\u003ePhoresy is likely the ancestral lifestyle of Astigmata, but it has been lost in (i) astigmatids inhabiting continuous habitats (such as soil or water), (ii) associated with non-nest-building insects; or (iii) nearly all vertebrates with overlapping generations, which enables maternal vertical transmission [1, 14]. Astigmata have experienced three major evolutionary events: (1) the ancestral oribatid life cycle (lacking a phoretic deutonymph) evolved to a life cycle with a specialized phoretic deutonymph (non-schizoglyphid Astigmata) or tritonymph (schizoglyphid Astigmata); (2) the deutonymphal lifecycle was then modified to permanently suppress the deutonymphal stage in several lineages, notably in Psoroptidia, a lineage of full-time (permanent) associates of birds and mammals, as these mites could effectively colonize new hosts via vertical transmission and other direct host-to-host contacts; (3) Pyroglyphidae, a lineage within Psoroptidia, transitioned to living in the nests of their hosts, thereby becoming secondarily free-living and using their former hosts for transport without forming phoretic deutonymphs [1, 17-19].\u003c/p\u003e\n\u003cp\u003eThe family Schizoglyphidae is the sister group to the remaining extant Astigmata [1]. This family is distinctive for retaining several plesiomorphic traits, including the relative position of the genital opening and attachment organ, as well as the structure of the attachment organ and gnathosoma compared to other astigmatid families [20]. This monotypic family includes a single observation of two specimens of \u003cem\u003eSchizoglyphus biroi\u003c/em\u003e Mahunka, 1978, found in New Guinea (Indonesia) on a tenebrionid beetle. Although this species exhibits the specific morphological adaptations characteristic of a phoretic heteromorphic deutonymph, it is probably a tritonymph\u0026nbsp;[16]. Evidence for this includes the presence of three pairs of genital papillae (as seen in oribatid tritonymphs and adults) instead of the two pairs found in deutonymphs and adults of other Astigmata. Despite substantial efforts to find this mite again on tenebrionid beetles, it has not been re-collected, suggesting that the original phoretic host (the beetle) may represent an incidental record rather than a true biologically relevant association. Thus, the true association of \u003cem\u003eSchizoglyphus biroi\u003c/em\u003e and the family it represents remains elusive.\u003c/p\u003e\n\u003cp\u003eAcariformes is one of the earliest diverging groups within the arachnids, with fossil evidence dating back to the early Devonian [21]. However, symbiotic associations with other organisms are mostly documented from the Cretaceous, when amber preserved these interactions \u003cem\u003ein situ\u003c/em\u003e [14, 22-23]. Crown-group Astigmata are estimated to have diverged between the Late Permian and the Early Triassic, while the stem-group likely originated in the Late Devonian to Carboniferous [13-14, 17, 22-23]. In the fossil record, Astigmata appears from the Cretaceous onward, becoming more abundant in the Cenozoic, with several instances of biotic associations [24-32].\u003c/p\u003e\n\u003cp\u003eHere, we report\u0026nbsp;the oldest amber association of an phoretic arthropod associated with its host preserved in Lebanese amber (Early Barremian, ~ 130 Ma): \u0026nbsp;16 mite deutonymphs belonging to the family Schizoglyphidae, phoretic on an alate termite \u003cem\u003eLebanotermes veltzi\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e(Figs. 1; and \u0026nbsp;2). To confirm this association and explore potential true hosts of \u003cem\u003eSchizoglyphus biroi\u003c/em\u003e, we also examined nests of modern termites in New Zealand and found schizoglyphids associated with \u003cem\u003eStolotermes\u003c/em\u003e, one of the earliest diverging termite lineages. The combined fossil and modern evidence suggests that schizoglyphid-termite associations represent the oldest known ongoing relationship between mites and their hosts. These findings shed light on the range of plesiomorphic features (including the gnathosoma and the attachment organ), present in the earliest crown-group Astigmata and reveal a highly conserved phoretic morphology that has persisted since the Early Cretaceous. This also advances our understanding of the temporal framework for the diverse mite-arthropod ecological associations observed today. \u0026nbsp;\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003eWe studied 16 mite specimens attached to the fossil holotype of termite \u003cem\u003eLebanotermes veltzi\u003c/em\u003e described in Engel \u003cem\u003eet al.\u003c/em\u003e [33]. The termite host which was preserved in a single amber piece (341 C\u0026ndash;T) from the Lower Barremian of Mdeyrij-Hammana, Caza (District) Baabda, Lebanon, coll. D. Azar. The amber piece is housed at the Natural History Museum of the Lebanese University, Faculty of Sciences II, Fanar. Other syninclusions derived originally from the same amber block include: the allotype of the chironomid dipteran \u003cem\u003eZiadeus kamili\u003c/em\u003e (341 B), \u0026nbsp;an aleyrodid hemipteran (341 A), and a ceratopogonid dipteran. The amber piece was trimmed and polished for microscopy, embedded in Canada balsam and placed in a cube of 1 mm thick microscopic glass slides. This permanent glass-amber preparation makes computed tomography not feasible for resolutions targeting the 150 \u0026micro;m-long mites.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSeveral microphotographs (Figs. 1A, C; 2A\u0026ndash;D, F; and 4A\u0026ndash;C) were acquired using the protocol described on this website (https://enrico-bonino.eu): a Sony a7R II mirrorless camera with a 208 mm tube lens, a Raynox DCR-150, microscope objectives Mitutoyo QV 2.5\u0026times; (or APO 20x), and a 110 mm tube lens with inversed Schneider Componon-S 50 mm/f2.8 lens for the whole specimen. This system (camera, tube-lens, and optics) is mounted on a MJKZZ Ultra Rail MINI V2, allowing movements in both vertical and horizontal planes. The illumination was provided by a cylindrical OGGLAB LED system DB 120EB. The entire panoramic image was assembled from two overlapping stacks, each composed of 53 frames. Images were captured in 16-bit RAW format with several steps between frames: 15 \u0026micro;m (with the Mitutoyo QV 2.5x), 2 \u0026micro;m (with the Mitutoyo 20x), and 95 \u0026micro;m (with Schneider-Componon lens). The frames were sub-stacked per group of eight images, stacked secondarily together and retouched in Helicon Focus (v.8.2, Professional) to remove undesired features (e.g. bubbles, dust, surfaces not in focus). Final image post-processing was done with Adobe Photoshop and Topaz DeNoise software. We also used a Zeiss Axiocam 208 mounted on an Axioimager A2 microscope, equipped with EC Epiplan 20x/0.4, 50x/0.55, and a W N-Achroplan 63x0.9 (water immersion) both in reflection and transmission light mode.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMultispectral microimaging was performed at a research platform IPANEMA (SOLEIL Synchrotron, Orsay, France, microscope magnification x20). Reflection and luminescence images emitted by the sample were collected in various spectral ranges using a setting coupling (1) an illumination device employing 16 different LED lights (from 365 up to 700 nm wavelength, CoolLED pE-4000) and (2) a light filter device placed in front of the camera detector (a wheel holding six interference band-pass filters collecting signal within in six spectral ranges from 435 to 935 nm). Out of the 96 produced illumination/detection couples, a selection of three couples enhancing morphological features of interest were combined into pseudo-coloured RGB. The stacking, alignment, image registration of the different couples, and production of pseudo-coloured RGB composites were performed using ImageJ. Pseudo-coloured RGB images were produced with red\u0026mdash;illumination 435 nm/detection 650 \u0026plusmn; 60 nm (luminescence), green\u0026mdash;470 nm/det. 650 \u0026plusmn; 60 nm, blue\u0026mdash;470 nm/det. 732 \u0026plusmn; 68 nm.\u0026nbsp;\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e1. \u003cstrong\u003eSystematic paleontology\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eClass \u003cstrong\u003eArachnida\u003c/strong\u003e Cuvier, 1812\u003c/p\u003e\n\u003cp\u003eSuperorder \u003cstrong\u003eAcariformes\u003c/strong\u003e Zachvatkin, 1952\u003c/p\u003e\n\u003cp\u003eOrder \u003cstrong\u003eSarcoptiformes\u003c/strong\u003e Reuter, 1909\u003c/p\u003e\n\u003cp\u003eSuborder \u003cstrong\u003eOribatida\u003c/strong\u003e Dug\u0026egrave;s, 1834\u003c/p\u003e\n\u003cp\u003eHyporder \u003cstrong\u003eAstigmata\u003c/strong\u003e Canestrini, 1891\u003c/p\u003e\n\u003cp\u003eFamily \u003cstrong\u003eSchizoglyphidae\u003c/strong\u003e Mahunka, 1978 (type genus \u003cem\u003eSchizoglyphus\u003c/em\u003e Mahunka, 1978)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003ePlesioglyphus\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;gen. n.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eZOOBANK CODE\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eType species.\u003c/strong\u003e\u003cem\u003e\u0026nbsp;Plesioglyphus lebanotermi\u003c/em\u003e gen. et sp. n.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eType material.\u0026nbsp;\u003c/strong\u003eHolotype (341E): phoretic tritonymph, specimen 1 (Figs. 2A; 3; and 5C), attached to alate specimen of the termite \u003cem\u003eLebanotermes veltzae\u003c/em\u003e Engel, Azar et Nel, 2011 (holotype), embedded in single amber piece (341 C\u0026ndash;T), the Lower Barremian of Mdeyrij-Hammana, Caza Baabda, Lebanon, coll. D. Azar, preserved at the Natural History Museum of the Lebanese University, Beirut. Paratypes: 15 phoretic deutonymphs (341 F\u0026ndash;T), same data.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eType locality and horizon.\u0026nbsp;\u003c/strong\u003eMdeyrij-Hammana, Caza (District) Baabda, Mount Lebanon Governorate, Lebanon, Lower Cretaceous, Lower Barremian.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDiagnosis.\u003c/strong\u003e \u003cem\u003eTritonymph\u003c/em\u003e. Subcapitular remnant large, with 2 pairs of short adoral setae (Fig. 3D). Palps free, long, 2-segmented; palp tarsus with at least 1 solenidion (Figs. 4F; and 5). Dorsal idiosoma sclerotized, punctate; sejugal furrow well developed. Progenital and anal opening well separated. Coxal fields III medially separated, distance between them is distinctly longer than the width of the trochanter III (Fig. 5B\u0026ndash;D). Anterior apodemes IV curved in the medial part, distinctly angular. Attachment organ large, with anterior suckers (\u003cem\u003ead\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e) and median suckers (\u003cem\u003ead\u003c/em\u003e\u003csub\u003e1+2\u003c/sub\u003e) well developed (Figs. 3B; .4D\u0026ndash;F; and 5B,\u0026ndash;D); conoids \u003cem\u003eps\u003c/em\u003e\u003csub\u003e1\u0026nbsp;\u003c/sub\u003eand \u003cem\u003eps\u003c/em\u003e\u003csub\u003e2\u003c/sub\u003e vestigial. Legs with typical segmentation (trochanter-tarsus). Tarsal empodial claws I-IV present, arising directly from tarsal apices (Fig. 5). Some tarsal setae foliate (Figs. 4F; and 5). \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRemarks\u003c/strong\u003e.\u0026nbsp;\u003cem\u003ePlesioglyphus\u003c/em\u003e belongs to Schizoglyphidae based on the presence of 2 pairs of adoral setae, well-developed palps, 4 pairs of genital setae, transversely elongated cuticular suckers formed by the fusion of pseudanal setae \u003cem\u003ep\u003csub\u003e1\u003c/sub\u003e+p\u003csub\u003e2\u003c/sub\u003e\u003c/em\u003e, and the anal opening situated between suckers \u003cem\u003ead\u003c/em\u003e\u003cem\u003e\u003csub\u003e1+2\u003c/sub\u003e\u003c/em\u003e.\u003cem\u003e\u0026nbsp;Plesioglyphus\u003c/em\u003e gen. n. is very similar to the extant genus \u003cem\u003eSchizoglyphus\u003c/em\u003e, but differs by the following: the gnathosoma is larger, reaching femora I (distinctly not reaching in \u003cem\u003eSchizoglyphus\u003c/em\u003e); the distance between suckers \u003cem\u003ead\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e and \u003cem\u003ead\u003c/em\u003e\u003csub\u003e1+2\u003c/sub\u003e is slightly smaller than the diameter of these suckers (slightly larger than the diameter of \u003cem\u003ead\u003c/em\u003e\u003csub\u003e1\u003c/sub\u003e in \u003cem\u003eSchizoglyphus\u003c/em\u003e); anterior apodemes IV are distinctly curved medially (curved only at tips in \u003cem\u003eSchizoglyphus\u003c/em\u003e).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEtymology.\u003c/strong\u003e The generic name is formed from two Greek stems, \u0026pi;\u0026lambda;\u0026eta;\u0026sigma;ί\u0026omicron;\u0026nu; (near, neighbouring) and \u0026gamma;\u0026lambda;ῠ́\u0026phi;\u0026omega; (to carve, cut out with a knife, engrave). The former stem is used in the formation of names in paleontology, while the latter stem is widely used to form names in astigmatid mites. Gender masculine.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003ePlesioglyphus lebanotermi\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;sp. n.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eZOOBANK CODE\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDescription.\u003c/strong\u003e \u003cem\u003eTritonymph\u003c/em\u003e. Gnathosoma large, subcapitular remnant subquadrate (width 1.2 times longer than length) (Fig. 3D). Palps 2-segmented. Palptarsus with at least one a seta and a solenidion \u0026omega;; solenidion \u0026omega; subequal or longer than palps (Figs. 3B\u0026ndash;C; 4F; and 5). Two pairs short adoral setae present (Fig. 3D). Gnathosoma (except for palps) situated under rostrum.\u003c/p\u003e\n\u003cp\u003eRostrum large, wide, rounded, without eyes. Dorsum with propodosomal and hysterosomal shields. Shields roughly punctate, separated by a distinct sejugal furrow. Dorsal setae not observed, except \u003cem\u003eh\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e on posterior hysterosoma.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eVentral side. Sternum present, long, nearly reaching anterior portions of apodemes II (specimen 12). Coxal apodemes II-IV with free ends, anterior apodemes IV distinctly curved medially, angular. Coxal fields I-IV open; coxal fields III well-separated medially, distance between them distinctly longer than width of trochanter III. Coxal setae not observed. Progenital opening nearly as long as base of legs III or IV, situated at level of trochanters IV, well-separated from attachment organ. Genital setae present, at least 4 pairs (Fig. 3B). Genital papillae not observed. Anal opening situated within attachment organ; distance between anal and progenital openings more than twice longer the length of progenital opening. Attachment organ large, almost as wide as body width. Suckers \u003cem\u003ead\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e and \u003cem\u003ead\u003c/em\u003e\u003csub\u003el+2\u003c/sub\u003e large, oval; \u003cem\u003ead\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e posterior to progenital opening, \u003cem\u003ead\u003c/em\u003e\u003csub\u003el+2\u003c/sub\u003e lateral to anal opening (Figs. 3B; 4D\u0026ndash;F; and 5B\u0026ndash;D). Distance between \u003cem\u003ead\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e and \u003cem\u003ead\u003c/em\u003e\u003csub\u003el+2\u003c/sub\u003e shorter than diameter of these suckers. Dorsoventral muscles of suckers \u003cem\u003ead\u003c/em\u003e\u003csub\u003el+2\u003c/sub\u003e well-developed (holotype). Alveolae \u003cem\u003eps\u003c/em\u003e\u003csub\u003e1\u003c/sub\u003e and \u003cem\u003eps\u003c/em\u003e\u003csub\u003e2\u0026nbsp;\u003c/sub\u003esituated between suckers \u003cem\u003ead\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e and \u003cem\u003ead\u003c/em\u003e\u003csub\u003el+2\u003c/sub\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eLegs short, thick, with typical set of segments. Empodial claws I-IV present, slightly shorter to tarsi (Fig. 5A,B). Tarsus I with seta \u003cem\u003ee\u003c/em\u003e, long, widened at tip and at least 1-2 other foliate setae, other setae spiniform. Tarsus II similar to tarsus I, but \u003cem\u003ee\u003c/em\u003e shorter. Tarsi III and IV with at least 6 foliate setae; 1 seta on tarsus IV longer than other (Figs. 4F; and 5C\u0026ndash;E). Tibiae I-II with 2 setae (\u003cem\u003egT\u003c/em\u003e and \u003cem\u003ehT\u003c/em\u003e) (Fig. 5,E), setae on tibiae III-IV not observed. Femora I-II with seta \u003cem\u003evF\u003c/em\u003e (Fig. 5C). Setation of tibiae III-IV, genua I-IV, femora III-IV and trochanters I-IV not observed. Tarsi I-II with 2 solenidia \u0026omega;. Tibiae I-II with 1 long solenidion \u0026phi; (its tip reaches tip of seta \u003cem\u003ee\u003c/em\u003e) (Figs. 4F; and 5). Tibia III with a single solenidion \u0026phi; (longer than combined length of genu and tarsus III) (Figs.3C; and 5). Genu I with one bacilliform solenidion \u0026sigma; (Fig. 5E). Other solenidia not observed.\u003c/p\u003e\n\u003cp\u003eMeasurements (n=3). Idiosoma 182\u0026ndash;220 long, 105\u0026ndash;110 wide. Prodorsum 68, width 95. Hysterosoma length 115. Gnathosoma 30\u0026ndash;35, width 23\u0026ndash;25; free palps 14\u0026ndash;15, gnathosomal solenidion \u0026omega; 14\u0026ndash;18. Length of attachment organ 50\u0026ndash;67, width 66\u0026ndash;70, \u003cem\u003ead\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e 17\u0026ndash;22 x 13\u0026ndash;19, \u003cem\u003ead\u003c/em\u003e\u003csub\u003el+2\u003c/sub\u003e 20\u0026ndash;28 x 17\u0026ndash;24. Legs I: length 46\u0026ndash;50, \u003cem\u003ee\u003c/em\u003e I 24\u0026ndash;30, \u0026omega; I 9\u0026ndash;11, \u0026phi; I 34\u0026ndash;36, \u0026sigma; I 10. Legs II: length 37, \u003cem\u003ee\u003c/em\u003e II 14\u0026ndash;22, \u0026phi; II 20.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRemarks.\u0026nbsp;\u003c/strong\u003eThe new\u0026nbsp;species differs from \u003cem\u003eSchizoglyphus biroi\u003c/em\u003e by long tibial solenidia \u0026phi; I\u0026ndash;II protruding the tips of respective tarsi (not protruding in \u003cem\u003eS. biroi\u003c/em\u003e). The following 16 phoretic deutonymphs were examined:\u003c/p\u003e\n\u003cp\u003eSpecimen 1 (341 E) (Fig. 2A; 3; and 5C) \u0026ndash; holotype, ventral. Legs and most of tarsal setae, gnathosoma (including adoral setae) and muscles are visible; a bubble inside the mite obscures observation.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSpecimen 2 (341 F) (Figs 2A; 4E\u0026ndash;F; and 5E) \u0026ndash; paratype, dorsolateral. Patterns on the dorsal shields and the posterior part of the attachment organs are well visible; the palp tarsus; legs I, right legs II -IV, anal opening, suckers \u003cem\u003ead\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e and \u003cem\u003ead\u003c/em\u003e\u003csub\u003el+2\u003c/sub\u003e are somewhat visible.\u003c/p\u003e\n\u003cp\u003eSpecimen 3 (341 G) (Fig. 2B) \u0026ndash; paratype, frontal view. Outlines of the gnathosoma, legs I, and sejugal furrow were observed.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSpecimen 4 (341 H) (Figs 2C; 4D; and 5D) \u0026ndash; paratype, ventral. The gnathosoma, suckers \u003cem\u003ead\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e and \u003cem\u003ead\u003c/em\u003e\u003csub\u003el+2\u003c/sub\u003e, \u003cem\u003eps\u003c/em\u003e\u003csub\u003e1\u003c/sub\u003e, anal and genital opening, some coxal apodema, and right tibial solenidion \u0026phi; II are visible; legs are somewhat visible as outlines.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSpecimens 5\u0026ndash;11 (341 G\u0026ndash;O), 13 (341 Q), 14 (341 R) (Fig. 2 E\u0026ndash;F) \u0026ndash; paratypes, ventral (5, 8), dorsolateral (6), lateral (7, 9, 13, 14), dorsal (10, 11). In specimens 10 and 11, hysterosomal and propodosomal shields (with rostrum), legs I-II, and sejugal furrow are somewhat visible. Other specimens are poorly visible.\u003c/p\u003e\n\u003cp\u003eSpecimen 12 (341 P)(Fig. 4C) \u0026ndash; paratype, ventral. Gnathosoma, legs I-II, genital and anal openings, apodemes I-II, attachment organ, legs III-IV are somewhat visible.\u003c/p\u003e\n\u003cp\u003eSpecimen 15 (341 R)(Fig. 4B) \u0026ndash; paratype, ventral.\u0026nbsp;The progenital and anal openings, suckers \u003cem\u003ead\u003c/em\u003e\u003csub\u003el+2\u003c/sub\u003e and left \u003cem\u003ead\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e, coxal areas are well visible. Legs I-III are somewhat visible.\u003c/p\u003e\n\u003cp\u003eSpecimen 16 (341 T) (Figs 2F; 4A; and 5A) \u0026ndash; paratype, dorsal. The dorsal shields, their sculpture, sejugal furrow, legs I-II with empodial claws are well visible. One dorsal seta (probably \u003cem\u003eh\u003c/em\u003e\u003csub\u003e3\u003c/sub\u003e) is somewhat visible.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2. Evidence for a true phoretic association preserved in amber\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe termite\u0026nbsp;\u003cem\u003eLebanotermes veltzae\u003c/em\u003e was preserved carrying 16 mites, positioned both dorsally (Fig. 1A\u0026ndash;B) and ventrally (Fig. 1C). Specimens 1\u0026ndash;4 are located at various points on or near the termite\u0026apos;s body, while specimens 5\u0026ndash;16 form a larger cluster over its abdominal wing area (Fig. 1B). Although some specimens were dislodged from the host (specimens 1-3, possibly during the process of amber entrapment, other specimens still remain attached to the wing membrane (specimens 4-16). Notably, specimen 16 has its pretarsal claws, equipped with foliate setae, in direct contact with the membrane of the right hindwing\u0026apos;s dorsal side (Figs. 2D, 2F; and 4A). The presence of 16 conspecific phoretic deutonymphs on the termite host, along with evidence of direct mite-host contact, supports the conclusion that this association is neither taphonomic nor random, but represents a true biological relationship.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.\u003c/strong\u003e \u003cstrong\u003eModern schizoglyphids are found in termite associations\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe earliest known crown-group astigmatid, \u003cem\u003eSchizoglyphus biroi\u003c/em\u003e, a heteromorphic phoretic tritonymph, was found on the tenebrionid beetle \u003cem\u003eDioedus tibialis\u003c/em\u003e (=\u003cem\u003eTagalus tibialis\u003c/em\u003e) in western New Guinea [35]. Despite extensive efforts to re-collect these mites from similar hosts (OConnor, pers. comm.), no further specimens were found, suggesting that this record was incidental rather than indicative of a true biological association. Another reported occurrence of \u0026quot;\u003cem\u003eSchizoglyphus\u003c/em\u003e sp.\u0026quot; on a scarabaeid larva from India [36] actually represents the genus \u003cem\u003eSancassania\u003c/em\u003e (family Acaridae). Here, we report the discovery of five deutonymphs of \u003cem\u003eSchizoglyphus sp.\u003c/em\u003e found in 2022 galleries of the New Zealand wetwood termite \u003cem\u003eStolotermes ruficeps\u003c/em\u003e (family Stolotermitidae), in a \u003cem\u003ePinus radiata\u003c/em\u003e log in Thames, New Zealand (Dickson Holiday Park, Tinker Trail, 37\u0026deg;06\u0026apos;42.7\u0026quot;S 175\u0026deg;31\u0026apos;22.1\u0026quot;E). The termite specimens were ethanol-washed from alate females, workers, and immature termites. These mites display several key character states that unequivocally place them within Schizoglyphidae: three-segmented palps (Fig. 6C), three pairs of genital papillae, five pairs of genital setae (Fig. 6B and D), the gnathosoma bearing adoral setae (Fig. 6B\u0026ndash;C), the cuticular suckers of the attachment organ are composed of\u0026nbsp;\u003cem\u003ep\u003csub\u003e1\u003c/sub\u003e+p\u003csub\u003e2\u0026nbsp;\u003c/sub\u003e\u003c/em\u003e(with alveoli), and the anal opening situated between suckers\u0026nbsp;\u003cem\u003ead\u003csub\u003e1+2\u003c/sub\u003e\u003c/em\u003e (Fig. 6B and D). These specimens represent a new species, which will be described in a future publication. Additionally, a different species of deutonymphal \u003cem\u003eSchizoglyphus sp.\u003c/em\u003e was previously collected from an alate queen of \u003cem\u003eStolotermes ruficeps\u003c/em\u003e in New Zealand, although these specimens were unfortunately lost (B. OConnor, pers. comm.). These new findings strongly suggest that schizoglyphids live inside termite nests, likely as inquilines, and use founder alate termites for dispersal from one nest to another.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe nests of social insects, such as termites, are stable, long-term habitats that provide abundant food resources, attracting a diverse array of inquiline organisms, including mites, flies, beetles, antlions, wasps, true bugs, silverfish, springtails, woodlice, harvestmen, pseudoscorpions, spiders, millipedes, and gastropods [9, 37-39]. Among these, \u003cstrong\u003eAstigmata\u003c/strong\u003e stands out as the most speciose lineage of termitophiles (see also Table 1 in supplementary materials) [40].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eRemarkably, very few astigmatans were reported on modern alate termites, compared to fossil record. We are confident interpreting it as a pure observation bias. First, because there are limitations in noticing them easily on alate hosts. Indeed, when mites are phoretic of alate reproducers of social insects (like the astigmatan \u003cem\u003eLemanniella\u0026nbsp;\u003c/em\u003esp. on the ant \u003cem\u003eMessor pergandei\u003c/em\u003e), the phoront often is hidden underneath the wings, at body contact (as in the bee \u003cem\u003eHalictus frontalis\u003c/em\u003e) [41]. Second, because amber has already proven to preserve phoretic associations which went nowadays discovered only after a fossil occurence had suggested their existence. That is the case of some larger wingless microarthropods (springtails), that were reported phoretic on diverse insect hosts in amber, including on alate social insects with the prediction that these interactions shall still exist [e.g. 38, 42-43]. Modern springtails were since indeed observed from the body of flies, tipulids and alate termites (N.R. pers. obs.). Indeed,\u0026nbsp;alates (reproducers) show a quite short lifespan compared to other casts of social insects: produced only at a certain time of year, and losing their wings right after nuptial flight, when flying away to form a new colony. In contrast, alate casts in amber are not uncommon (both males and females), with resin patches forming along the barks mostly above ground level, trapping the flying individuals. This is illustrated by amber termite-mite records:\u0026nbsp;unidentified mites and Mesostigmata were respectively found on extinct Euisoptera [38]\u0026nbsp;from Jordanian amber with a similar Early Barremian age as Lebanese amber [44]\u0026nbsp;(~ 130 \u0026nbsp;Ma)\u0026nbsp;and a Cenomanian\u0026nbsp;(~\u0026nbsp;100Ma)\u0026nbsp;termite-like roach from Burmese amber [45]. Both records indeed involve alate adults as well (female in post-nuptial flight in the second case). \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAstigmatan heteromorphic nymphs, including \u003cem\u003ePlesioglyphus lebanotermi\u003c/em\u003e, have a highly specialized attachment organ (Figs. 4\u0026ndash;6) serving for attachment and hitchhiking on hosts [14] [46]. The success of a deutonymph attachment depends on its location on the host\u0026apos;s body, to both prevent detachment by the host and minimize interference with the host\u0026apos;s locomotion abilities [47]. Among the 16 individuals of \u003cem\u003eP. lebanotermi\u003c/em\u003e, most specimens were found on wings, on areas of overlap between hindwings (sp. 5\u0026ndash;15, Fig. 1; and\u0026nbsp;2D), or between hindwing and forewings (sp. 16, Figs. 1; 2F; and 5E) suggesting that the fossil deutonymphs were in this position during their transport.\u003c/p\u003e\n\u003cp\u003eAs astigmatid heteromorphic deutonymphs lack a mouth, oral feeding is not possible. Still feeding as a parasite can occur during phoresy via attachment organ suckers, anus or genital papillae [1,48]. Inside the nest, certain mites (such as \u003cem\u003eAustralhypopus\u003c/em\u003e sp.) may provide sanitary functions by feeding on dead termite corpses [5-6]. During phoresy, mites can potentially impede the mobility of their hosts when the mite loads are high [49-50]; but, in general, phoresy is harmless to them. The significant advantage gained by phoronts is linked to an increased dispersal distance, allowing them to exploit new food resources or access different hosts. In social insects, dispersal is enhanced during periods of colony swarming through attachment to alate reproducers. At swarming, alate termites can travel distances of over one kilometre [51]. The observed\u0026nbsp;~ 130 \u0026nbsp;Ma old\u0026nbsp;association of \u003cem\u003ePlesioglyphus lebanotermi\u003c/em\u003e with \u003cem\u003eLebanotermes veltzae\u003c/em\u003e is here most likely to represent phoretic commensalism, while non-phoretic stages of the mites likely live inside termite nests as commensals.\u003c/p\u003e\n\u003cp\u003eFour distinct superfamilies of Astigmata are associated with four different termite lineages (Fig. 7; Table 1\u0026nbsp;in supplementary materials). Among these, the superfamily Acaroidea is the most commonly reported, with 13 genera and 21 species identified from members of the Rhinotermitidae (including \u003cem\u003ePsammotermes hypostoma\u003c/em\u003e, \u003cem\u003eReticulitermes flavipes\u003c/em\u003e, and \u003cem\u003eCoptotermes formosanus\u003c/em\u003e) and one Termitidae species (\u003cem\u003eCornitermes cumulans\u003c/em\u003e) (Fig. 7; Table 1\u0026nbsp;in supplementary materials). The superfamily Histiostomatoidea has been recorded on several rhinotermitid species, while a species of the superfamily Hemisarcoptoidea has been found on \u003cem\u003ePsammotermes hypostoma\u003c/em\u003e (Rhinotermitidae). The frequent association of Acaroidea with termites is expected, as this superfamily is species-rich, comprising 562 species [1]. However, despite comparable species diversity (576 species), there are fewer reports of phoretic associations involving Histiostomatidae. Hemisarcoptoidea is a smaller group, with 144 species. All three of these mite superfamilies are associated with Neoisopteran termites, which are believed to have diverged approximately 110 million years ago (with the split between Termopsidae and Rhinotermitidae occurring around 85 Ma [51], Fig. 7).\u003c/p\u003e\n\u003cp\u003eIn contrast, both modern and fossil schizoglyphoid associations, including those reported here, are found on termite lineages that diverged much earlier\u0026mdash;from the Early Barremian (126 Ma for Stolotermitidae) to the Late Lias (185 Ma for Lebanotermes, Euisoptera; [52]). This distribution may suggest a degree of specialization among phoretic astigmatid mites for specific termite lineages, possibly selected through co-occurrence over geological time and maintained to the present day (Fig. 7). However, the limited number of schizoglyphid occurrences currently prevents any definitive assessment of this pattern.\u003c/p\u003e\n\u003cp\u003eSchizoglyphid phoretic nymphs retain several plesiomorphic traits when compared to other astigmatid mites. These include: (i) A gnathosomatic remnant with two pairs of adoral setae and relatively long palps, whereas other astigmatids lack adoral setae and have reduced palps; (ii) Three pairs of genital papillae (though this was not observed in \u003cem\u003ePlesioglyphus lebanotermes\u003c/em\u003e) and four or more pairs of genital setae, while other astigmatids typically have two pairs of genital papillae and only one pair of genital setae; (iii) Pseudanal setae \u003cem\u003ep\u003csub\u003e1\u003c/sub\u003e+p\u003csub\u003e2\u003c/sub\u003e\u003c/em\u003e are fused into large, transversely elongated cuticular suckers, unlike the small, rounded suckers seen in most astigmatids; (iv) The anal opening is positioned more posteriorly than in most astigmatids, situated at the level of \u003cem\u003ead\u003csub\u003e1+2\u003c/sub\u003e\u003c/em\u003e, rather than \u003cem\u003ead\u003csub\u003e3\u003c/sub\u003e\u003c/em\u003e as is typical in other astigmatids. There are interesting similarities between \u003cem\u003ePlesioglyphus\u003c/em\u003e the phoretic nymphs of the genus \u003cem\u003eLevantoglyphus\u003c/em\u003e (family Levantoglyphidae), recently reported from Lebanese amber without host information [13]. Both genera share a well-developed gnathosomal remnant with long palps and long terminal solenidia (\u0026omega;), indicating the importance of host-seeking behaviour in both lineages. However, \u003cem\u003eLevantoglyphus\u003c/em\u003e possesses rudimentary chelicerae, enabling food shredding in non-phoretic stages, whereas modern schizoglyphids from New Zealand retain a mouth and pharynx, suggesting that the reduction of functional mouthparts in astigmatid phoretic nymphs was a gradual evolutionary process.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003ePlesioglyphus lebanotermi\u003c/em\u003e displays all the synapomorphies of modern schizoglyphid mites and documents the existence of non-feeding, phoretic heteromorphic nymphs in astigmatid mites from the Early Cretaceous (~130 Ma). This fossil represents the earliest known crown-group Astigmata with a confirmed phoretic association with termites\u0026mdash;a relationship that has persisted into modern times. Its placement among living mites will allow for precise calibration of molecular clock phylogenies. In contrast, the transitional deutonymphs of the extinct stem-group family \u003cem\u003eLevantoglyphidae\u003c/em\u003e, whose host associations remain unknown, provide a less precise calibration for molecular clock phylogenies. As these deutonymphs belong to a stem-group lineage of Astigmata, their use in calibrating molecular clocks is limited to a broader and less specific range compared to \u003cem\u003ePlesioglyphus lebanotermi\u003c/em\u003e.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe oldest known biotic association of arthropods preserved in amber, dating to approximately 130 million years ago, involves Astigmata, a group of mites specialized in phoresy, which were found attached to a winged termite. This discovery represents the earliest known instance of phoretic mites associated with an arthropod. The mites belong to the genus \u003cem\u003ePlesioglyphus\u003c/em\u003e n. gen. of the family Schizoglyphidae, an early-diverging lineage of Astigmata currently recognized from a single described species. Remarkably, the plesiomorphic features of these ancient mites \u0026mdash;such as long palps and a large subcapitulum\u0026mdash; have been highly conserved over 130 million years. In the Early Cretaceous of Lebanon, these schizoglyphids coexisted with other extinct Astigmata (\u003cem\u003eLevantoglyph\u003c/em\u003e\u003cem\u003eus)\u003c/em\u003e that also exhibited plesiomorphic mouthparts. However, unlike these extinct relatives, the schizoglyphids described here are the earliest known crown-group Astigmata to display strictly non-feeding, heteromorphic nymphs. This finding reveals a specialized phoretic relationship with their termite host, suggesting that \u003cem\u003ePlesioglyphus\u003c/em\u003e functioned as an inquiline in its feeding stages, and dispersed on winged termites via phoresy during non-feeding, heteromorphic stages. This association, which has persisted into modern times, highlights the long-standing evolutionary relationship of Schizoglyphidae with eusocial insects \u0026mdash; a connection that has been largely overlooked. Our findings show the remarkable diversity and evolutionary persistence of modern Schizoglyphidae, which continue to exhibit phoresy on termites, reflecting their ancient and ongoing relationship with these insects.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eField studies were conducted in accordance with local legislation. As for Lebanon, the amber collecting was performed after obtention by Pr. Dany Azar of the necessary authorisations from the Municipality of Hammana, the Lebanese Ministry of Power, Mining direction; and the National Council for Scientific Research - Lebanon (National central public institution in charge of science policy-making under the authority of the President of the Council of Ministers).\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the reported information was used with the consent of their owners.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data supporting these findings are included in this article.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was supported by a BELSPO BRAIN-be federal Belgian grant (B2/202/P1/PARADI2S) and by the EU NextGenerationEU through the Recovery and Resilience Plan for Slovakia under the project No. 09I03-03-V04-00439. P.B.K. and V.B.K. were supported by the Ministry of Science and Higher Education of the Russian Federation within the framework of the Federal Scientific and Technical Program for the Development of Genetic Technologies for 2019-2027 (agreement No. 075-15-2021-1345, unique identifier RF----193021X0012). This paper is a contribution to the activity of the laboratory \u0026lsquo;Advanced Micropalaeontology, Biodiversity and Evolution Researches (AMBER)\u0026rsquo; led by DA at the Lebanese University.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConceptualization and methodology by N.R., H.S., P.K.. Formal analysis and visualization by H.S., N.R., V.K., P.K., E.B. and J.K.. Investigation and writing of the original draft by H.S., N. Robin, P.K. and E.B.. Writing\u0026nbsp;- Review \u0026amp; Editing by V.K. and D.A.. Resources by D.A. and P.K.. Project administration and supervision by N. R. and D.A. All authors confirm their authorship and approve the final version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank Dmitry Vorontsov for technical help, Barry OConnor for his record of the additional modern deutonymphal \u003cem\u003eSchizoglyphus\u003c/em\u003e, Michael Engel for valuable scientific advice on termites, and Peter Vr\u0026scaron;ansk\u0026yacute; for providing amber samples with evidence of mite syninclusions along with cockroaches. \u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003cp\u003e1.\u0026nbsp;Seeman OD, Walter DE. Phoresy and mites: More than just a free ride. 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J Biol Control\u003cem\u003e.\u0026nbsp;\u003c/em\u003e2023;37(4):268\u0026ndash;270.\u0026nbsp;\u003ca href=\"https://doi.org/10.18311/jbc/2023/36348\"\u003ehttps://doi.org/10.18311/jbc/2023/36348\u003c/a\u003e \u0026nbsp;\u003c/p\u003e\n\u003cp\u003e37. Aguiar NO, B\u0026uuml;hrnheim PF. Phoretic pseudoscorpions associated with flying insects in Brazilian Amazonia.\u0026nbsp;J Arachnol. 1998;26(3):452\u0026ndash;59.\u003c/p\u003e\n\u003cp\u003e38. Vr\u0026scaron;ansk\u0026yacute;\u0026nbsp;P, \u0026Scaron;m\u0026iacute;dov\u0026aacute; L, Sendi H, Barna P, Mueller P, Ellenberger S, et al.\u0026nbsp;Parasitic cockroaches indicate complex states of earliest proved ants. Biologia. 2019;74:65\u0026ndash;89.\u0026nbsp;\u003ca href=\"https://doi.org/10.2478/s11756-018-0146-y\"\u003ehttps://doi.org/10.2478/s11756-018-0146-y\u003c/a\u003e\u003c/p\u003e\n\u003cp\u003e39. Magowski WL. Discovery of the fist representative of the mite subcohort Heterostigmata (Arachnida, Acari) in the Mesozoic Siberian amber. Acarologia. 1994;35:229\u0026ndash;41.\u003c/p\u003e\n\u003cp\u003e40. Wang C, Powell JE, O\u0026rsquo;Connor BM. Mites and nematodes associated with three subterranean termite species (Isoptera: Rhinotermitidae). Fla Entomol. 2002;85(3): 499\u0026ndash;506.\u0026nbsp;\u003ca href=\"https://doi.org/10.1653/0015-4040(2002)085%5b0499:MANAWT%5d2.0.CO;2\"\u003ehttps://doi.org/10.1653/0015-4040(2002)085[0499:MANAWT]2.0.CO;2\u003c/a\u003e\u003c/p\u003e\n\u003cp\u003e41. Engel MS.\u0026nbsp;New augochlorine bees (Hymenoptera: Halictidae) in Dominican amber, with a brief review of fossil Halictidae. J Kans Entomol Soc. 1996; 69(4):334\u0026ndash;45.\u0026nbsp;\u003ca href=\"https://doi.org/10.3897/zookeys.29.257\"\u003ehttps://doi.org/10.3897/zookeys.29.257\u003c/a\u003e\u003c/p\u003e\n\u003cp\u003e42 Boucot AJ, Poinar Jr GO. Fossil behavior compendium. Boca Raton, USA; CRC Press: 2010.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e43. Gr\u0026uuml;nemaier, M. Phoretic springtail (Collembola: Sminthuridae) on a false blister beetle (Coleoptera: Oedemeridae) in Eocene Baltic amber. Palaeodiversity. 2016;9(1): 9-13.\u0026nbsp;\u003ca href=\"https://doi.org/10.18476/pale.v9.a2\"\u003ehttps://doi.org/10.18476/pale.v9.a2\u003c/a\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e44. Azar D, G\u0026egrave;ze R, Acra F, Penney D. 2010Lebanese amber. In:\u0026nbsp;Penney D., editor. Biodiversity of fossils in amber from the major world deposits. Manchester, UK: Siri Scientific Press; 2010. p. 271\u0026ndash;98.\u003c/p\u003e\n\u003cp\u003e45.\u0026nbsp;Vr\u0026scaron;ansk\u0026yacute; P, Koubov\u0026aacute; I, Vr\u0026scaron;ansk\u0026aacute; L, Hinkelman J, K\u0026uacute;dela M, K\u0026uacute;delov\u0026aacute; T, et al. Early wood-boring \u0026lsquo;mole roach\u0026rsquo;reveals eusociality \u0026ldquo;missing ring.\u0026rdquo; AMBA projekty. 2019;9(1):1\u0026ndash;28.\u003c/p\u003e\n\u003cp\u003e46.\u0026nbsp;O\u0026rsquo;Connor BM. Evolutionary ecology of Astigmatid Mites. Annu Rev Entomol. 1982;27(1):385\u0026ndash;409.\u0026nbsp;\u003ca href=\"https://doi.org/10.1146/annurev.en.27.010182.002125\"\u003ehttps://doi.org/10.1146/annurev.en.27.010182.002125\u003c/a\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e47. Houck MA, OConnor BM. Ecological and Evolutionary Significance of Phoresy in the Astigmata. Annu Rev Entomol. 1991;36:611\u0026ndash;36.\u003c/p\u003e\n\u003cp\u003e48. Houck MA, Cohen AC. The potential role of phoresy in the evolution of parasitism: Radiolabelling (tritium) evidence from an astigmatid mite. Exp Appl Acarol 1995:19 (12): 677\u0026ndash;694.\u0026nbsp;\u003ca href=\"https://doi.org/10.1007/bf00052079\"\u003ehttps://doi.org/10.1007/bf00052079\u003c/a\u003e\u003c/p\u003e\n\u003cp\u003e49. Phillipsen WJ, Coppel HC. \u003cem\u003eAcotyledon formosani\u003c/em\u003e sp. n. associated with the Formosan subterranean termite, Coptotermes formosanus Shiraki (Acarina: Acaridae-Isoptera: Rhinotermitidae). J Kans Entomol Soc. 1977;50(3):399\u0026ndash;409.18. Eraky SA. \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e50. Luong LT, Penoni LR, Horn CJ, Polak M. Physical and physiological costs of ectoparasitic mites on host flight endurance. Ecol Entomol. 2015;40:518\u0026ndash;24.\u0026nbsp;\u003ca href=\"https://doi.org/0.1111/een.12218\"\u003ehttps://doi.org/0.1111/een.12218\u003c/a\u003e\u003c/p\u003e\n\u003cp\u003e51. Messenger MT, Mullins AJ New flight distance recorded for \u003cem\u003eCoptotermes formosanus\u003c/em\u003e (Isoptera: Rhinotermitidae). Fla Entomol.\u0026nbsp;2005;88(1): 99\u0026ndash;100.\u0026nbsp;\u003ca href=\"https://doi.org/10.1653/0015-4040(2005)088%5b0099:NFDRFC%5d2.0.CO;2\"\u003ehttps://doi.org/10.1653/0015-4040(2005)088[0099:NFDRFC]2.0.CO;2\u003c/a\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e52. Jouault C, Legendre F, Grandcolas P, Nel A. Revising dating estimates and the antiquity of eusociality in termites using the fossilized birth\u0026ndash;death process. Syst Entomol. 2021,46:592\u0026ndash;610.\u0026nbsp;\u003ca href=\"https://doi.org/10.1111/syen.12477\"\u003ehttps://doi.org/10.1111/syen.12477\u003c/a\u003e\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Royal Belgian Institute of Natural Sciences","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Isoptera, Lower Cretaceous, Barremian, social insects, Lebanese amber","lastPublishedDoi":"10.21203/rs.3.rs-5389108/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5389108/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAmong minute-sized and wingless arthropods, astigmatid mites stand out for their diverse range of symbiotic associations (parasitic, neutral and mutualistic), with both invertebrate and vertebrate hosts. When inhabiting discontinuous and ephemeral environments, astigmatid mites adapt their life cycle to produce a phoretic heteromorphic nymph. When feeding resources are depleted, phoretic nymphs disperse to new habitats through phoresy, attaching to a larger animal which transports them to new locations. This dispersal strategy is crucial for accessing patchy resources, otherwise beyond the reach of these minute arthropods. In Astigmata, the phoretic nymph is highly specialized for dispersal, equipped with an attachment organ and lacking a mouth and pharynx. Despite the common occurrence of phoretic associations in modern mites, their evolutionary origins remain poorly understood. Among Astigmata, the family Schizoglyphidae represents an early derivative lineage with phoretic tritonymphs; however, our knowledge of this family is limited to a single observation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHere, we report the oldest biotic association of arthropods fossilised in amber (~130 Ma, Lebanon): an alate termite with 16 phoretic deutonymphs of Schizoglyphidae\u003cem\u003e(Plesioglyphus lebanotermi\u003c/em\u003e gen. et sp. n.). The mites are primarily attached to the membranes of the host’s hindwings, using their attachment organs, pretarsal claws and tarsal setae. Additionally, we report new modern phoretic tritonymphs of this same family, on one of the earliest lineages of termites. These data collectively indicate that schizoglyphid-termite associations represent the oldest continuous mite-host associations. Notably, schizoglyphid mouthparts retain a distinct mouth and pharynx, absent in modern Astigmata.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe discovery of Schizoglyphidae mites in Lebanese amber represents the oldest known continuous association between acariform mites and their hosts. This finding demonstrates the long-term evolutionary significance of phoresy in Astigmata, evidencing a relationship sustained for over 130 Ma. It indicates that these early mites lived inside termite nests as inquilines and used alate termites for dispersal. This ancient association offers key insights into the coevolution of both mites and termites, highlighting a potential for the future discoveries of similar mites. This fossil —a stem-group Astigmata— is important for the accurate calibration of acariform mite phylogenies, advancing our understanding of these mites evolutionary history.\u003c/p\u003e","manuscriptTitle":"The oldest continuous association between astigmatid mites and termites preserved in Cretaceous amber reveals the evolutionary significance of phoresy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-20 15:14:01","doi":"10.21203/rs.3.rs-5389108/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"e3e26972-ff42-44cc-b801-ffd9caaf4e69","owner":[],"postedDate":"November 20th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-11T03:38:10+00:00","versionOfRecord":[],"versionCreatedAt":"2024-11-20 15:14:01","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5389108","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5389108","identity":"rs-5389108","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}