First records of three Pergamasinae species (Acari, Mesostigmata, Parasitidae) from Slovakia

We report three mite species, Pergamasus ( Thenargamasus ) instatutus Athias-Henriot, 1967, Pergamasus ( Pergamasus ) laminarius Witaliński, 1971, and Holoparasitus ampullaris Witaliński, 1994, from Slovakia for the first time. The genera Pergamasus Berlese, 1903 and Holoparasitus Oudemans, 1936 are predatory mites living mainly in soil and rotting organic matter of the Holarctic region, with a centre of distribution in Europe. Until now, P. instatutus was only recorded in Austria and Hungary, while P. laminarius and H. ampullaris were known only from Poland.


Introduction
The family Parasitidae Oudemans, 1901 is divided into two subfamilies, Parasitinae Oudemans, 1901 andPergamasinae Juvara-Balş, 1972. The mites of the subfamily Pergamasinae are mostly soil-inhabiting predators, but they can be found in many other habitats as nests of birds and small mammals, carcasses of small mammals, in mosses, compost, etc. (Ambros 1993;Fenďa and Mašán 2003;Fenďa and Schniererová 2005). By far the most species-32 of the genus Pergamasus-have been recorded in Austria (Wilmann 1953;Athias-Henriot 1967;Schmölzer 1995). Significantly fewer species have been recorded in any other country. Other countries with large numbers of recorded Pergamasus species are Romania and Slovakia, both with 14 species recorded (Juvara-Balş 1976; our own records). Most species of Holoparasitus are known from southern and central Europe, but this genus has also been recorded from other parts of Europe, such as the British Isles and the Baltic coast (Karg 1971;Hyatt 1987;Witaliński 1994aWitaliński , 1994bWitaliński , 2017aWitaliński 2000, 2006).

Methods
We collected mites from the nest of Clanga pomarina (Brehm, 1831) and from soil using three methods-soil samples, soil samples sieving, and pitfall traps. The nests of C. pomarina were sampled in 1999 by a student Slávka Siryová for her diploma thesis in cooperation with the organization Raptor Protection of Slovakia. Approximately 200 cm 3 of nest lining was removed from nests after nesting season, when the nests were empty, and this substrate was processed similarly to soil samples; we used Tullgren funnels for extracting mites from these samples. We used only one nest containing mites for this study. We stored the mites in 70% ethanol and separated them from other soil arthropods using a Leica EZ4 stereomicroscope. Subsequently, we mounted the mites on permanent microscopic slides using a chloralhydrate medium. We observed the mites under a Zeiss Axioscope 5 compound microscope. We took the photographs with a Zeiss Axiocam 208 color camera using Zeiss Labscope 3 software. The photographs were stacked in Adobe Photoshop 22.0. We made the map using a software QGIS 3.16. The localities are shown in Figure 1. The abbreviation "ibid." is used for "ibidem" and indicates repetitive data. The voucher specimens are deposited in Zoological Collection of Department of Zoology, Faculty of Natural Sciences, Comenius University of Bratislava, Slovakia.

Pergamasus (Thenargamasus) instatutus Athias-Henriot, 1967
Pergamasus instatutus Athias-Henriot 1967: 686;figs. 31, 68, 124, 278, 284, 317, 326.   Identification. Only females are known. The original description provided by Athias-Henriot (1967) is concise but contains the most important and characteristic features. The most distinctive feature that distinguishes this species from all other Pergamasus species is the form of the endogynium ( Fig. 2A): spherules are round, a stipule is deeply divided, and branches are diverged into a T-shape; trabecules are long and straight with short fingerlike or sharp protrusions that are usually located in the middle part of the trabecules and apically. Gnathotectum with five prongs and several small additional denticles between lateral prongs. Anterolateral margins of epigynium almost straight (Fig. 2B). Lyrifissures iv2 are short and located closer to the anterior margin of a paragynial shield than to setae st4 (Fig. 2C).
This species is very similar to Pergamasus (Thenargamasus) barbarus (Berlese, 1904) ( Fig. 2D-F), but P. instatutus differs by the form of the endogynium. The stipule of P. instatutus only slightly exceeds the spherules while the stipule of P. barbarus is usually twice as long as the spherules. Branches of the stipule in P. barbarus are diverged into a Y-shape instead of a T-shape and there are no teeth between the branches, unlike in the species P. instatutus. In addition, the trabecules of P. barbarus are bent backwards apically, possess long sharp protrusions resembling thorns pointing adaxially, and small sharp protrusions can be present at the level of the spherules.   (Berlese, 1904) but is considerably smaller, the length of idiosoma is 890-1200 µm in males of P. laminarius and 1130-1400 µm in males of P. brevicornis. The females can be distinguished on the basis of the endogynium and epigynium (Fig. 3). The endogynium possesses several small, well-discernible denticles on the wall of the endogynial sac, and the absence of adaxial processes of trabecules is characteristic (Fig. 3A, B). The epigynium (Fig. 3D) is relatively short and lacks the teeth on its inner (dorsal) surface that are present in P. brevicornis (Fig. 3E).
Males differ mainly by the shape of chelicerae and armature of tibia II (Fig. 4). The fixed digit is wide, thinner apically and with a dorsal hump (Fig. 4A) but not a tooth as in P. brevicornis (Fig. 4E). The longitudinal apohysis of tibia II is proximally higher and triangular and without transverse stripes (Fig. 4B). The males of P. laminarius and P. brevicornis can be confused due to a characteristic saddle-shaped apophysis on genu II (Fig. 4C, G). Witaliński (1971) pointed out characteristic endings of setae pv1 on tibia and genu IV. However, the endings of these setae are similar in several species of the beklemischevi species-group (Fig. 4 D, H).

Discussion
Pergamasus instatutus was previously found in Austria, near Graz and in northern Hungary, which are almost 300 km apart (Athias-Henriot 1967). The new records are a little further north but basically somewhere between these two places. However, these are the first records of this species since Athias-Henriot (1967) described it. Her material was collected in 1944 and 1952. Pergamasus instatutus is probably widespread in deciduous forests in the lower mountains of Austria, Hungary, and Slovakia. Intensive research on soil mites can still provide information on wider distributional range of P. instatutus, but it noteworthy that this species has not been found in Romania, despite intensive research in this country (Juvara-Balş 1976). Although we found 65 individuals, we did not find males, and so the males remain unknown. Pergamasus laminarius was previously found only in southern Poland, near the Slovak border (Witaliński 1971;Gwiazdowicz and Sznajdrowski 1999). This species was found in soil samples often together with the species P. brevicornis. However, while P. brevicornis is common and widespread in Europe, P. laminarius is a rarer species distributed in the Carpathians. We found it in a wide range of habitats from xerophile meadows to beech forests. Only two records of Holoparasitus ampullaris were known, both from Poland. Micherdziński (1969) found it in Kasprowy Wierch in the Tatra Moutains for the first time, but he published it as H. excisus. Witaliński (1972) recorded it from the environs of Krakow in southern Poland, also as H. excisus. He later described a new species based on those individuals (Witaliński 1994b). Kasprowy Wierch, where this species was found for the first time, is only a few meters from the Slovak border, so it was no surprise to us that we found H. ampullaris also on the Slovak side of the Tatras. While in Poland it was found in mosses and hummus from rock crevices, as well as in litter and mosses in young coniferous forests, we found it in soil and litter of young mixed forest. Although only nine individuals are known from Poland, we found up to 47 individuals, and this species was present in 11 of the 20 samples taken from our study area.