Antispilina ludwigi Hering, 1941 (Lepidoptera, Heliozelidae) a rare but overlooked European leaf miner of Bistorta officinalis (Polygonaceae): new records, redescription, biology and conservation Author Nieukerken, Erik j. van Naturalis Biodiversity Center, PO Box 9557, NL- 2300 RA Leiden, The Netherlands; nieukerken @ naturalis. nl Author Wullaert, Steve Weg naar Bijloos 15, B- 3530 Houthalen, Belgium; sw. demijnen @ gmail. com - www. bladmineerders. be Author Lee, Bong-Woo Korea National Arboretum, 415 Gwangneungsumokweon-ro, Soheul, Pocheon, Gyeonggi-Prov., 11186 South Korea; eucosma @ korea. kr Author Bryner, Rudolf Bergluftweg 19, CH- 2505 Biel, Switzerland; rbryner @ bluewin. ch text Nota Lepidopterologica 2021 2021-04-23 44 99 121 http://dx.doi.org/10.3897/nl.44.63848 journal article http://dx.doi.org/10.3897/nl.44.63848 2367-5365-44-99 2DE821FE01E9401C971B6AC3829A8204 DFD2A417E5E15452AED128BEBED3C8AA Antispilina ludwigi Hering Antispilina ludwigi Hering, 1941:19. Holotype ♂, Germany : [Hessen] "Dillkreis, Im Feuerhack bei Waldaubach", emerged 20 March 1941. Mines 29 July 1940, Polygonum bistorta [MfN] [examined] Antispilina ludwigi ; Ludwig 1952 : 24; Grabe 1955 : 100; Buhr 1964 : 37 [Sachsen]; Wojtusiak 1976 : 9 [Poland, key, description, genitalia + adult ill.]; Razowski 1978 : 91. [Poland, redescription, ill]; Lastuvka 1998 : 17 [Czech Republic, checklist]; Gaedike and Heinicke 1999 : 43 [Germany, catalogue]; Liska et al. 2000 : [Czech Republic]; Borkowski 2003a : 88 [Poland]; Borkowski 2003b : 113 [Poland, photographs adults and mines]; Ivinskis 2004 : 36 [Lithuania]; Buszko and Palka 2016 : 245 [Poland]; Gaedike et al. 2017 : 24 [Germany, catalogue]; Buszko and Nowacki 2017 : 17 [Poland, catalogue]; van Nieukerken 2018: https://fauna-eu.org/cdm_dataportal/taxon/6b71a06e-5ecb-48b6-84f0-45462a9d4604 [Europe, catalogue]; Lastuvka et al. 2018 : 121-122 [Czech Republic, colour illustrations]; Bryner in Lepiforum 2003-2021: http://www.lepiforum.de/lepiwiki.pl?Antispilina_Ludwigi [Switzerland, photographs all stages, genitalia and mines]; Werkgroep Bladmineerders 2020: http://www.bladmineerders.be/nl/content/antispilina-ludwigi-m-hering-1941 [Belgium, photographs mines, larvae], De Prins and Steeman 2003-2021: https://projects.biodiversity.be/lepidoptera/species/5448/ [Belgium, catalogue]. Diagnosis. Antispilina ludwigi is a small completely dark grey-black moth, including the forewing fringe, with a narrow silver fascia at one-third and two opposite silver spots at two-thirds, resembling Antispila species. The small size (4-5 mm wingspan) and the dark colour separate A. ludwigi from the larger Antispila species (4.8-9 mm), that also have a distinct fringe line and white forewing fringe. Holocacista rivillei (Stainton, 1855) is about the same size, but the fascia is usually divided, it has also a white fringe, and the male foretibia bears a small epiphysis and it is not so dark. The latter species occurs in Mediterranean habitats with wild or cultivated grapevine ( Vitis vinifera L.), whereas A. ludwigi is confined to poor grasslands and moors, mostly in mountains. A key was provided by van Nieukerken et al. (2018) . The leafmine is the only blotch mine known on Bistorta officinalis , but early mines may sometimes be confused with the reddish circular patches caused by the fungus Ramularia bistortae Fuckel ( Mycosphaerellaceae , Ascomycota ) ( Ellis 2020 ) (e.g. Fig. 26 ). Description. Male (Figs 1 , 24 , 25 ). Forewing length 2.1-2.4 mm, wingspan ca. 4.5-5.0 mm. Head (Figs 3-5 ): frons with smooth scaling, bronzy-fuscous with brassy reflection; vertex bronzy-fuscous. Antenna reaching 2/3 of forewing, with 18-19 segments, basally covered with fuscous scales; scape pale brown ventrally with pecten consisting of 3 or 4 hairs. Labial palpus pale fuscous, comprising 2 palpomeres; maxillar palpi minute, 1-articled. Haustellum as long as head capsule. Thorax and tegulae brassy with metallic reflections, may appear silvery in some lighting. Legs dark grey, no white spots, epiphysis absent. Forewing almost jet-black, apically with brassy reflections, a silvery white, narrow outwardly oblique fascia at 1/5 to 1/4, usually constricted in middle; a silvery white small triangular dorsal spot at 1/2, and a squarish costal spot slightly beyond dorsal one; fringe grey with brassy reflection, fringe line absent. Hindwing and fringe dark grey. Abdomen greyish black. Female (Fig. 2 ). Forewing length 2.0-2.3 mm, wingspan ca. 4.2-5.0 mm. Abdomen apically pointed. Figures 1, 2. Antispilina ludwigi , male and female. Switzerland, Bern, Rueeggisberg . Scale bar 1 mm. Photographs R. Bryner. Figures 3-5. Antispilina ludwigi , male adult, morphological details, whole body slide EvN4448. 3. Foreleg. 4 . Head and mouthparts, lp: labial palp with 2 palpomeres; mp: maxillary palp, 1-articled. 5. Head and antenna with 19 segments. Scale bars: 100 µm (1, 2), 200 µm (3). Photographs E.J. van Nieukerken. Venation (Fig. 6 ). Forewing: Sc poorly visible. R unbranched, a separate vein, to costa, but a persistent trachea connecting R with Rs+M+CuA. Rs+M+CuA ending in 4 rather indistinct branches, Rs1+2 to costa, Rs3+4 to termen, M and CuA to dorsum. Hindwing with Sc+R to costa, poorly visible, Rs+M with 3 branches, Rs indistinct, to costa, 2 branches of M to termen and dorsum; CuA a separate vein to dorsum; no visible Anal vein. Figure 6. Antispilina ludwigi , male venation, slide EvN4676. Photograph E.J. van Nieukerken. Male genitalia (Figs 7 - 13 ). Total length ca 530 µm . Vinculum well sclerotized, long, narrowest in middle, as long as valva or longer. Tegumen bilobed. Valva subtriangular with blunt apex, length twice width, stalked pecten ca. 2/3 from base of valva, with ca. 12-13 blunt sensilla, the number on each valva may differ. Transtilla with long sublateral processes and slightly bilobed posterior process. Juxta weakly sclerotized, three-pronged anteriorly. Phallus cylindrical, ca. 400 µm long, without cornuti, distal part slightly swollen, about as long as vinculum; phallocrypt spinose. Figures 7-9. Antispilina ludwigi , male genitalia, slide EvN4448. 7. Ventral view, single exposure. 8. Stacked image from four individual exposures. 9. Detail, stack from two exposures. Scale bars 100 µm (7, 8), 50 µm (9). Photographs E.J. van Nieukerken. Figures 10-13. Antispilina ludwigi , male genitalia, Germany, Hessen: Dillkreis. Drawn in glycerin. 10, 11. whole genitalia, phallus removed, lateral and ventral view. 12, 13. phallus, lateral and ventral view. Scale bar 100 µm . Art work Bong-Woo Lee. Female genitalia (Figs 14-17 ). Tip of oviscapt dorsoventrally flattened, with five cusps in total. Posterior apophyses slightly longer than anterior apophyses. Eighth sternum membranous, except narrow sclerotized region in distal part. Eighth tergite sclerotized, posterior part dorsally elevated, anterior end crooked, middle part projected anteriorly. Guy wire arising from middle part of vestibulum, as long as vestibulum. Vestibulum membranous with a pair of sclerotized rods. Figures 14-17. Antispilina ludwigi , female genitalia, Germany, Hessen: Dillkreis. Drawn in glycerin. 14, 15. whole genitalia, lateral and ventral view. 16. vestibulum, ventral view. 17. Oviscapt, detail. Scale bars 200 µm , 50 µm (17). Art work Bong-Woo Lee. Larva (Figs 18-22 , 35 ). Body white, prothoracic plates and head pale brown. Prosternum and pronotum with ovate plates (sclerites) in what we assume are the 4th and 5th instars (see van Nieukerken and Eiseman 2020 ); earlier instars not examined in detail), other segments and prothorax outside the plates as well covered with small transverse swellings. Head width 4th instar ca 400 µm . Legs and prolegs absent. The 4th instar cuts out the flat cocoon or shield from pieces of the leafmine. Figures 18-23. Antispilina ludwigi , immature stages. 18, 19. Details of slide mounted caterpillar, probably 4th instar, Poland, RMNH.INS.12423; scale bars 100 µm . 20. Caterpillar, final feeding instar (4th) in opened mine, Switzerland, St-Imier, 2.viii.2017. 21. Non feeding instar (5th) from opened shield, Switzerland, Saignelegier , 5.ii.2018. 22. Caterpillar walking with shield, Switzerland, St-Imier, 3.viii.2017. 23. Pupa in opened cocoon, Switzerland, Saignelegier , 4.v.2018. Photographs E.J. van Nieukerken (18, 19), R. Bryner. Pupa (Fig. 23 ). Not studied in detail. Biology. Host plant. Snake-root (or Common Bistort), Bistorta officinalis Delarbre (= Polygonum bistorta L., Persicaria bistorta (L.) Samp., Bistorta major Gray) ( Polygonaceae ). Figures 24-25. Antispilina ludwigi , live male, Switzerland, Saignelegier , 4.v.2018. Photographs R. Bryner. Leafmine (Figs 26 - 35 ). The exact oviposition site has not been determined, but in most cases this must be close to the midrib (87% of the French mines, n=251), or close to another large vein. The larva first makes a circular blotch close to the vein with the frass glued to the upper and under epidermis, resulting in a round red-brownish spot on both leaf surfaces. From there the mine enlarges into an elongated blotch mine, usually between lateral veins towards leaf margin; the blotch is further transparent and whitish in colour. From there the frass is more scattered, blackish, partly in clumps and no longer concentrated throughout the rest of the mine. The outer edges of the mine are irregularly eaten out and do not contain any frass. In many cases the mine is not larger than ca 1 cm, in thin leaves this can be larger. At the end the caterpillar returns to the round spot and makes there an elongate to circular leaf cut from both epidermal layers (reinforced by the frass) of ca. 3 mm long (Fig. 32 ). There are often several mines per leaf, and mines regularly occur as "twins" at both sides of the midrib (e.g. Fig. 31 ). In some cases several mines converge into large communal mines, where several larvae may feed next to each other (Fig. 35 ). Leaves with many mines are very conspicuous, even from a distance. Figures 26-33. Antispilina ludwigi , leafmines and larvae on Bistorta officinalis . 26, 27. France, Mt. Mezenc , 29.vii.2009. 28, 29. France, Puy-de-Dome , Res. Nat. Chastreix-Sancy, resp. Roc de Courlande and Fontaine Salee , 13.vii.2017. 30-33. Belgium, Rocherath, Naturschutzgebiet der Holzwarche, 23.vi.2017. Photographs E.J. van Nieukerken (26-29), S. Wullaert (30-33). The larva crawls away with the shield (Fig. 22 ) and seeks a shelter, where it will moult inside the shield into the non-feeding 5th instar and later pupate. The larva probably attaches the shield to some plant material (Fig. 36 ) as in the other genera in the Holocacista -group (van Nieukerken and Eiseman 2020 ). Larvae were found from June to September (own data; Liska et al. 2000 ; Borkowski 2003b ; Buszko and Palka 2016 ), earlier at lower altitudes, in the mountains most larvae were active in July. Reared adults always emerged after hibernation the next spring between March and June; one adult was swept from the hostplant on 27.v.2020 in the locality St. Imier at 1095 m (Switzerland). We assume that there is only one generation, with an extended period of larval feeding, and adults flying from April to June depending on altitude. Figures 34-37. Antispilina ludwigi , leafmines and larvae on Bistorta officinalis and habitat. 34-36. Switzerland, Saignelegier , 4.viii.2017. 37. Habitat in Switzerland, Jura, St-Imier, probably in flying time of Antispilina ludwigi , taken 21.vi.2009. Photographs R. Bryner. Parasitoids. We only reared three specimens of Pnigalio sp. ( Eulophidae , identified by Hannes Baur) and one Chalcidoidea , unidentified to lower taxon (identified by Seraina Klopfstein). All emerged in February from cocoons that were taken indoors in January (locality Saignelegier , Jura, Switzerland). No parasitoids emerged from the extensive breedings of French material. Distribution (Fig. 40 ). Central Europe: recorded from Belgium, France, Switzerland, Germany, the Czech Republic, Poland and Lithuania. The species has previously only been found a few times: in Germany in a small area in Hessen and Nordrhein-Westfalen ( Hering 1941 ; Ludwig 1952 ; Grabe 1955 ) and Sachsen ( Buhr 1964 ); in Poland it was known from the Sudety mountains ( Wojtusiak 1976 ; Razowski 1978 ; Buszko and Nowacki 2000 ; Borkowski 2003a , b ), but later also found in the lowlands in the South-East ( Buszko and Palka 2016 ), one record from the Czech Republic, also from the Sudety range ( Liska et al. 2000 ) and from southern Lithuania ( Ivinskis 2004 ). Here we report its occurrence for the first time from France (Massif Central: Auvergne, Cevennes and Jura), Switzerland (Jura and Alps) and Belgium (Ardennes). It is very likely that A. ludwigi also occurs further east in Belarus, Ukraine or Russia, where Microlepidoptera and especially leafmining Lepidoptera have only been collected very sparsely. Habitat (Figs 37 - 39 ). Most of our own records were obtained in mountainous bogs, poor wet meadows, often along streams and montane heathland where the host is abundant. This is often patchy habitat with shrubs and sometimes scattered larger trees. The attacked hostplants often are relatively large and leaves with mines are usually hidden among the vegetation. Smaller plants in mowed grasslands rarely had mines. The following plants were common in many of the localities in the Massif Centrale: Arnica montana L., Betula pubescens Ehrh., Calluna vulgaris (L.) Hull, Cicerbita alpina (L.) Wallr., Cytisus oromediterraneus Rivas Mart. et al., Dianthus gratianopolitanus Vill., Genista pilosa L., G. tinctoria L., Gentiana lutea L., G. pneumonanthe L., Meum athamanticum Jacq., Potentilla erecta (L.) Raeusch., Rumex acetosa L., Sanguisorba officinalis L., Succisa pratensis Moench, Trollius europaeus L., Vaccinium myrtillus L., and V. uliginosum L. Figures 38, 39. Habitats of Antispilina ludwigi . 38 (top). Belgium, Rocherath, Naturschutzgebiet der Holzwarche, 24.vi.2017. 39 (bottom). Ardeche , Sainte-Eulalie, Suc de la Lauziere , 23.vii.2017. Photographs: A. Rauw (38), E.J. van Nieukerken (39). Buszko and Palka (2016) found the species in lowland habitats in ' Molinietum-caeruleae ' meadows, between 170 and 215 m. Our records are from mountains between 450 and 1650 m elevation and also most published records are from mountains in similar elevational range. Figure 40. Distribution of records of Antispilina ludwigi , including all literature records. DNA barcodes. Nine specimens were barcoded, representing populations in Poland, France and Belgium. All cluster within Barcode Identification Number BOLD:AAW5935, with an average distance of 0.36% and a maximum distance of 0.81%. (https:/dx.doi.org/10.5883/BOLD:AAW5935; dataset https:/dx.doi.org/10.5883/DS-ANTILUDW. The nearest neighbour, at a distance of 9.38%, is an unnamed heliozelid from Costa Rica. Remark. There are several specimens in collections with paratype labels, such as the three cited here (and see Lepiforum e.V. (2008-2021)), but as Hering (1941) cited a single specimen in the description, the type material consists only of a holotype, and no paratypes. Material examined. 21 ♂♂ 19 ♀♀ , cocoons. All specimens are reared from Bistorta officinalis , except where indicated. France • 2 ♂♂ 1 ♀ ; Ardeche , Mt Mezenc , SW slopes; 44.90835°N , 4.18781°E ; alt. 1652 m ; 29.vii.2009 ; E.J. van Nieukerken leg.; emerged 03.iv- 03.v.2010 ; EventId: EvN no 2009032-K; Genitalia slides: EvN4448 ♂ (whole body slide), EvN4676 ♂ , EvN4677 ♀ ; RMNH.INS.24448, RMNH.INS.24676, RMNH.INS.24677. • 5 ♂♂ 4 ♀♀ , cocoons; same data; emerged 03.iv- 06.v.2010 ; no registry numbers; RMNH. • 2 cocoons, 1 with prepupa; Ardeche , Sainte-Eulalie, Bois des Seigneurs; 44.86449°N , 4.1827°E ; alt. 1455 m ; 23.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017053-K; RMNH.INS.16908.• 4 ♂♂ , 10 cocoons (plus exuviae); Ardeche , Sainte-Eulalie, Suc de la Lauziere , S.; 44.84695°N , 4.17325°E ; alt. 1483 m ; 23.vii.2017 ; E.J. van Nieukerken leg.; emerged 02-22.iv.2018 ; EventId: EvN no 2017051-K; RMNH.INS.16584- RMNH.INS.16587, RMNH.INS.16907. • 2 ♀♀ ; Puy-de-Dome , Monts Dore, Chastreix-Sancy, Roc de Courlande, S.; 45.52231°N , 2.7688°E ; alt. 1339 m ; 13.vii.2017 ; E.J. van Nieukerken leg.; emerged 02.iv.2018 ; EventId: EvN no 2017034-K; RMNH.INS.16567- RMNH.INS.16568. • 2 ♂♂ 4 ♀♀ 1 ad, 10 cocoons (plus exuviae); Puy-de-Dome , Monts Dore, Chastreix-Sancy, Res. Nat, Fontaine Salee ; 45.51214°N , 2.79792°E ; alt. 1338 m ; 13.vii.2017 ; E.J. van Nieukerken leg.; emerged 29.iii- 09.iv.2018 ; EventId: EvN no 2017035-K; RMNH.INS.16569- RMNH.INS.16575, RMNH.INS.16902. • 3 ♂♂ ; Puy-de-Dome , Monts Dore, Col de la Croix de St Robert, SE, along road; 45.55873°N , 2.84282°E ; alt. 1416 m ; 16.vii.2017 ; E.J. van Nieukerken leg.; emerged 03-12.iv.2018 ; EventId: EvN no 2017039-K; RMNH.INS.16578- RMNH.INS.16580. • 1 ♂ ; Puy-de-Dome , Monts Dore, Vallee de Chaudefour, Res. Nat., upper valley E; 45.52916°N , 2.83496°E ; alt. 1346 m ; 16.vii.2017 ; E.J. van Nieukerken leg.; emerged 04.iv.2018 ; EventId: EvN no 2017037-K; RMNH.INS.16577. • 4 cocoons; Puy-de-Dome , Monts Dore, Chastreix-Sancy, Roc de Courlande, S.; 45.52231°N , 2.7688°E ; alt. 1339 m ; 13.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017034-K; RMNH.INS.16901. • 9 cocoons; Puy-de-Dome , Monts Dore, Col de la Croix de St Robert, SE, along road; 45.55873°N , 2.84282°E ; alt. 1416 m ; 16.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017039-K; RMNH.INS.16905. • 1 cocoon; Puy-de-Dome , Monts Dore, Vallee de Chaudefour, Res. Nat., upper valley E; 45.52916°N , 2.83496°E ; alt. 1346 m ; 16.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017037-K; RMNH.INS.16904.• 1 ♀ , 5 cocoons (plus exuviae); Puy-de-Dome , St. Alyre-es-Montagne , Lac de St. Alyre, S. shore; 45.37424°N , 2.94417°E ; alt. 1224 m ; 14.vii.2017 ; E.J. van Nieukerken leg.; emerged 09.iv.2018 ; EventId: EvN no 2017036-K; RMNH.INS.16576, RMNH.INS.16903. Germany • 2 ♂ 1 ♀ [labelled as paratypes ]; Hessen , Dillkreis , Waldaubach , im Feuerhack ; 50.666474°N , 8.132751°E ; alt. 570 m ; 29.vii.1940 ; A. Ludwig leg.; emerged 10.iii.1941 ; Genitalia in vial; MfN . Switzerland • 1 ♂ 1 ♀ ; Bern , Rueeggisberg ; 46.7307°N , 7.45849°E ; alt. 1400 m ; 07.viii.2017 ; R. Bryner leg.; emerged 04-06.v.2018 ; Genitalia slide ♂ 2018-031; RBC. • 2 ♀♀ ; Bern , St-Imier ; 47.12879°N , 6.98725°E ; alt. 1095 m ; 02.viii.2017 ; R. Bryner leg.; emerged 04-08.v.2018 ; RBC . • 1 ♀ ; same locality; 27.v.2020 ; R. Bryner & D. Bolt leg.; day catch; collection Daniel Bolt , Domat / Ems .• 1 ♂ 2 ♀♀ ; Jura , Saignelegier ; 47.23961°N , 7.04089°E ; alt. 1000 m ; 04.viii.2017 ; R. Bryner leg.; emerged 04-12.v.2018 ; Genitalia slide ♀ 2018-032; RBC. Material examined: larvae and leafmines (in collection). All collected from Bistorta officinalis . Belgium • 100 larvae , 146 mines; Liege , Aldringen , Thommen ; 50.22563°N , 6.02781°E ; alt. 475 m ; 24.vi.2017 ; WB leg.; SWC . • 2 larvae , 2 mines; Liege , Aldringen, Ulf; 50.2147°N , 6.03125°E ; alt. 462 m ; 24.vi.2017 ; WB leg.; SWC. • 16 larvae , 16 mines; Liege , Buellingen , Kleinfuellenbach ; 50.39425°N , 6.32834°E ; alt. 644 m ; 24.vi.2017 ; WB leg.; SWC. • 3 mines; Liege , Emmels, Deidenberg; 50.31102°N , 6.14507°E ; alt. 481 m ; 24.vi.2017 ; WB leg.; SWC. • 3 larvae (used for DNA analysis, 2 destructively), leafmines; Liege , Rocherath , Naturschutzgebiet der Holzwarche ; 50.4238°N , 6.314457°E ; alt. 585 m ; 23.vi.2017 ; Steve Wullaert leg.; larval slide; RMNH.INS.30919, RMNH.INS.30927, RMNH.INS.30928, RMNH.INS.43338. • 105 mines; same locality data; WB leg.; SWC. • 4 mines; same locality; 19.viii.2017 ; WB leg.; SWC . France • several mines; Ardeche , Mt Mezenc , Croix de Boutieres ; 44.90018°N , 4.18277°E ; alt. 1512 m ; 29.vii.2009 ; E.J. van Nieukerken leg.; EventId : EvN no 2009033-K; RMNH.INS.42771. • 3 larvae , many mines; Ardeche , Mt Mezenc , SW slopes; 44.90835°N , 4.18781°E ; alt. 1652 m ; 29.vii.2009 ; E.J. van Nieukerken leg.; EventId: EvN no 2009032-M/H/K; RMNH.INS.17942- RMNH.INS.17944 (larvae); RMNH.INS.42667- RMNH.INS.42668 (leafmine samples). • 1 larva , 3 mines; Ardeche , Sainte-Eulalie, Bois des Seigneurs ; 44.86449°N , 4.1827°E ; alt. 1455 m ; 23.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017053-M/H/K; RMNH.INS.31178, RMNH.INS.43887. • many mines; Ardeche , Sainte-Eulalie, Pre du Bois , W of Suc de la Lauziere ; 44.84959°N , 4.16323°E ; alt. 1469 m ; 23.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017052-H/K; RMNH.INS.43472- RMNH.INS.43473. • 2 larvae , mines; Ardeche , Sainte-Eulalie, Suc de la Lauziere , S. ; 44.84695°N , 4.17325°E ; alt. 1483 m ; 23.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017051-M/H.K; RMNH.INS.30932- RMNH.INS.30933, RMNH.INS.43470- RMNH.INS.43471. • 2 larvae , mines; Puy-de-Dome , St. Alyre-es-Montagne , Lac de St. Alyre , S. shore; 45.37424°N , 2.94417°E ; alt. 1224 m ; 14.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017036-M/H/K; RMNH.INS.30929- RMNH.INS.30930, RMNH.INS.43463, RMNH.INS.43465, RMNH.INS.43872. • 8 mines; Puy-de-Dome , Monts Dore, Chastreix-Sancy, Roc de Courlande , S. ; 45.52231°N , 2.7688°E ; alt. 1339 m ; 13.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017034-H/K; RMNH.INS.43868, RMNH.INS.43869. • 2 larvae , many mines; Puy-de-Dome , Monts Dore, Chastreix-Sancy, Res. Nat, Fontaine Salee ; 45.51214°N , 2.79792°E ; alt. 1338 m ; 13.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017035-M/K/H; RMNH.INS.30931, RMNH.INS.30939; RMNH.INS.43462, RMNH.INS.43870, RMNH.INS.43871. • 1 larva , several mines; Puy-de-Dome , Monts Dore, Col de la Croix de St Robert , SE , along road; 45.55873°N , 2.84282°E ; alt. 1416 m ; 16.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017039-K; RMNH.INS.31004, RMNH.INS.43466, RMNH.INS.43875. • 2 mines; Puy-de-Dome , Monts Dore, Vallee de Chaudefour , Res. Nat., upper valley E; 45.52916°N , 2.83496°E ; alt. 1346 m ; 16.vii.2017 ; E.J. van Nieukerken leg.; EventId: EvN no 2017037-K; RMNH.INS.43873 . Poland • 29 larvae [ethanol, some destructively extracted, larval slide], Dubienka ; 51.00998°N , 23.92238°E ; alt. 175 m ; 02.vii.2004 ; K. Palka leg.; EventId : EvN no 2004905-M; RMNH.INS.11854, RMNH.INS.11855, RMNH.INS.12423 . • 15 larvae [ethanol, some destructively extracted], Poleski Nat. Park : Bagno Bubnow ; 50.94514°N , 22.637°E ; alt. 287 m ; 01.vii.2004 ; K. Palka leg.; EventId : EvN no 2004904-M; RMNH.INS.11852, RMNH.INS.11853 . Switzerland • 5 larvae , Bern , Cortebert ; 47.15431°N , 7.12766°E ; alt. 1260 m ; 04.viii.2017 ; R. Bryner leg.; RBC. • leafmines; Bern , Saicourt ; 47.2555°N , 7.16569°E ; alt. 920 m ; 02.viii.2017 ; R. Bryner leg.; RBC. • leafmines; Bern , St-Imier ; 47.17653°N , 6.94997°E ; alt. 1000 m ; 02.viii.2017 ; R. Bryner leg.; RBC. • leafmines; Bern , Tramelan ; 47.24232°N , 7.06144°E ; alt. 1010 m ; 04.viii.2017 ; R. Bryner leg.; RBC. • leafmines; Jura , Lajoux ; 47.28891°N , 7.1292°E ; alt. 930 m ; 04.viii.2017 ; R. Bryner leg.; RBC. • leafmines; Jura , Le Bemont ; 47.25269°N , 7.02997°E ; alt. 970 m ; 04.viii.2017 ; R. Bryner leg.; RBC. • leafmines; Jura , Montfaucon ; 47.2819°N , 7.08989°E ; alt. 850 m ; 04.viii.2017 ; R. Bryner leg.; RBC. • leafmines; Neuchatel , La Brevine ; 46.97567°N , 6.58592°E ; alt. 1030 m ; 05.viii.2017 ; R. Bryner leg.; RBC. • leafmines; Neuchatel , Le Cerneux-Pequinot ; 47.00417°N , 6.66141°E ; alt. 1060 m ; 05.viii.2017 ; R. Bryner leg.; RBC. • leafmines; Vaud , Ste-Croix ; 46.84122°N , 6.47154°E ; alt. 1090 m ; 05.viii.2017 ; R. Bryner leg.; RBC . Additional online observations. All observed on Bistorta officinalis . Belgium • 8 larvae, 30 mines; Liege , Butchenbach, Schwalm; 19.vii.2018; leg. Evert Van de Schoot & Ward Tamsyn; 50.4980, 6.2562; https://waarnemingen.be/observation/160366513/. • 1 larva, 10 mines; Liege , Butchenbach; Schwalm; 19.vii.2018; leg. Evert Van de Schoot; 50.4898, 6.2714; https://waarnemingen.be/observation/160463202/. • 3 mines; Liege , Malmedy, Fagne de la Polleur; 22.vii.2017; leg. Wim Declercq; 50.5105, 6.0744; https://waarnemingen.be/observation/141930607/. • 13 larvae, 16 mines; Liege , Rocherath, Vallee de la Holzwarche; 24.vi.2017; leg. WB; 50.3961°N , 6.3318°E ; https://waarnemingen.be/observation/140955240/. • 2 larvae, 2 mines; same locality and date; leg. Chris Steeman & Ben Steeman; 50.4151°N , 6.3283°E ; https://waarnemingen.be/observation/140964609/. • 1 mine; same locality; 22.vii.2017; leg. Wim Declercq; 50.4172°N , 6.3268°E ; https://waarnemingen.be/observation/141930615/. • 2 larvae, 2 mines; same locality; 22.vii.2017; leg. Wim Declercq; 50.4240°N , 6.3145°E ; https://waarnemingen.be/observation/141930621/. • 1 mines; same locality; 14.vii.2018; leg. Jurgen Dewolf & Ruben Recour; 50.4237°N , 6.3147°E ; https://waarnemingen.be/observation/159940474/. • 20 larvae, 40 mines; same locality; 23.vii.2018; leg. Wouter Mertens; 50.4209°N , 6.3199°E ; https://waarnemingen.be/observation/160514341/. • 14 larvae, 34 mines; same locality; 23.vii.2018; leg. Chris Steeman; 50.4237°N , 6.3145°E ; https://waarnemingen.be/observation/160831314/. • 4 larvae, 25 mines; same locality; 29.vii.2018; leg. Regis Nossent; 50.4236°N , 6.3146°E ; https://waarnemingen.be/observation/160702446/. • 1 larva, 1 mines; same locality; 06.ix.2018; leg. WB; 50.4233°N , 6.3141°E ; https://waarnemingen.be/observation/162347416/. • 2 larvae, 7 mines; same locality; 08.viii.2019; leg. WB; 50.4233°N , 6.3152°E ; https://waarnemingen.be/observation/178051781/. • 16 larvae, 26 mines; same locality; 10.viii.2019; leg. WB; 50.4209°N , 6.3182°E ; https://waarnemingen.be/observation/179079003/. • 1 larva, 8 mines; same locality; 08.viii.2020; leg. WB; 50.4246°N , 6.3145°E ; https://waarnemingen.be/observation/203156253/. • 19 larvae, 76 mines; Liege , Waimes, Rue du Ponce ; 24.vii.2018; leg. Chris Steeman & Steve Wullaert; 50.4387°N , 6.1095°E ; https://waarnemingen.be/observation/160817401/. • 3 larvae, 4 mines; same locality; 09.viii.2020; leg. Eric Wille; 50.4384°N , 6.1093°E ; https://waarnemingen.be/observation/198140856/. • 14 larvae, 15 mines; Luxembourg, Vance, Marais de Vance; 02.vii.2017; leg. Wim Declercq; 49.6719°N , 5.6807°E ; https://waarnemingen.be/observation/141082788/. • 7 larvae, 8 mines; Luxembourg, Volaiville, Vallee de la Sure ; 20.vi.2020; leg. WB; 49.8658°N , 5.6846°E ; https://waarnemingen.be/observation/194673026/. France • 1 vacated mine; Jura, Les Rousses; 08.viii.2020; leg. Wim Declercq; 46.4963°N , 6.0759°E ; https://observation.org/observation/199070630/.