Deanophlebia kala (Harker) Finlay, 2019

Finlay, K. J., 2019, Deanophlebia: A new genus of south-eastern Australian mayflies and subsequent redefinition of the subgenus Nousia (Australonousia) (Ephemeroptera: Leptophlebiidae), Zootaxa 4668 (1), pp. 30-50 : 30-50

Deanophlebia kala (Harker) comb. nov.

Atalophlebia kala Harker ( Harker 1954)

Nousia sp. AV3 ( Dean 1999)

Deanophlebia kala (Harker) comb. nov.

Types. Holotype. Atalophlebia kala Harker, Lake Albina, Mount Kosciusko , New South Wales, 2 II 1929, ♂ imago pinned, R.J. Tillyard. Labels: a) “ Atalophlebia kala ”, b) “Abdomen in slide cabinet”, c) Mt. Kosciusko, Lake Albina 2.2.29, R.J. Tillyard ”, d) “ Type ”, e) Brit. Mus. 1937-408 ”. Paratypes. Lake Cootapatamba , Mount Kosciusko, New South Wales, 27 I 1930, R. J.Tillyard, ♀ imago, subimagos [not seen]. Held at the NHM .

Distribution. Southern New South Wales, Eastern Victoria.

Diagnosis. Male Imago. Dimensions: body length 9.3–13.1 (11.1±1.25, 15); forewing length 10.3–12.8 (11.8±0.80, 14); hindwing length 1.9–2.9 (2.52±0.36, 13); forewing hindwing ratio 4.07–5.79 (4.74±0.52, 13). Gen- eral body colour tan to dark brown. Head dark brown. Antennae: pedicel and scape dark brown, flagellum tan brown. Ocelli: three; black with white-grey inserts; laterals larger than medial. Eyes: upper eye size large, sometimes contiguous; ES=0.00 – 0.11 (0.06±0.04, 10); upper lobes orange-tan brown, lower lobes grey-black. Thorax: pronotum dark brown; mesonotum tan brown with darker brown markings. Legs: forelegs with femora tan brown, apices darker; tibiae and tarsi yellow; total leg length 9.8–11.4 (10.9±0.57, 10); forelegs with seven segments; length ratios 1.00: 1.51: 0.05: 0.45: 0.45: 0.37: 0.15 (2.76±0.21, 10); tarsal claws of a pair similar, each apically hooked with an opposing hook ( Fig. 1 ). Wings: forewing ( Fig. 2 ): membrane hyaline, pterostigmal region opaque, longitudinal and crossveins brown, anal veins yellow; costal and subcostal crossveins present in proximal and distal halves of wing; costal space with 5–9 (12) cross veins proximally, 12–20 (12) distally (some anastomosed), subcostal space with 5–8 (12) cross veins proximally, 11–16 (12) distally. Three bullae present on veins Sc, R 2 and R 4+5, MA forked at half to just over half the distance from wing base to margin, MP 2 connected to MP 1 and CuA, CuA and CuP linked by crossvein, ICu 1 recurved to join CuA and not linked to CuA-CuP crossvein, ICu 2 recurved to join ICu 1, CuA and CuP linked by crossvein, ICu 1 and ICu 2 parallel as wing margin approached, CuP strongly recurved and linked by crossvein to A 1. Hindwing ( Fig. 3 ): mostly hyaline, costal space washed with yellow and very slightly opaque; costal margin slightly convex at midlength immediately preceding shallow concavity, costa joins subcosta at approximately four-fifths wing length; no crossveins in proximal half of costal space, 5–8 (11) in distal space, 6–8 (11) throughout subcostal space.Abdomen ( Fig. 4 ): tan brown, darker posteriorly, usually with golden markings on each segment forming a broken stripe midline, two golden coloured maculae flanking each midline marking anteriorly, segments eight and nine usually darker, segment ten usually light, colour and pattern variable. Genitalia ( Figs 5–7 ): forceps three segmented ( Fig. 5 ); tan brown; terminal segment angular, sometimes indented, about the same length as middle segment; penes tan brown; fused along entire length dorsally and ventrally, lobes often contiguous ( Figs 5 & 6 ); outer lateral margins bulbous; two lobes distinct, each lobe pointing outwards, triangular processes present apically on each lobe, two pairs of internal spines seen from dorsal side, not always apparent ( Fig. 7 ). Caudal filaments: three; tan brown; apex of each segment washed dark brown giving a banded appearance; terminal filament longer than cerci. Female imago. Dimensions: body length 9.6–12.6 (11.2±1.00, 15); forewing length 11.3–15.6 (12.8±1.02, 14); hindwing length 2.2–3.0 (2.62± 0.25, 13); forewing hindwing ratio 4.27–5.61 (4.93±0.44, 13). General patterning and colouring similar to male. Eyes: grey-black, separated on meson of head by a distance about four times maximum width of eye. Legs: total leg length shorter than male, total foreleg length 7.6–9.4 (8.3±0.59, 10); forelegs with six segments, leg length ratios 1.00: 1.43: 0.21: 0.18: 0.15: 0.13 (2.69±0.23, 10). Wings: forewing ( Fig. 8 ): colour and venation similar to male except wings with more crossveins; costal space with 7–8 (11) cross veins proximally, 16–20 (11) distally (some anastomosed), subcostal space with 7–9 (11) crossveins proximally, 14–16 (11) distally. Hindwing ( Fig. 9 ): no crossveins in proximal half of costal space, 6–8 (10) in distal half, 7–10 (10) throughout subcostal space. Abdomen: sternum seven with small genital extension; sternum nine moderately cleft ( Fig. 10 ). Egg. Ovoid; polar cap absent, chorion with large and small circular shaped protuberances over surface, pattern variable ( Figs 11 & 12 ). Male and female subimago. Body lengths: ♂ 9.3–10.6 (9.7 ±0.61, 10); ♀ 10.0–13.7 (11.5±1.05, 15). General colour and markings similar to imago. Wings uniformly greybrown, opaque. Forelegs of male not elongated. Genitalia not fully developed. Mature nymph ( Fig. 13 ). Body lengths: ♂ 10.0–12.5 (11.1±0.86, 10); ♀ 11.5–14.9 (13.0±1.00, 16). General colour tan with darker brown markings. Head: prognathous; tan to dark brown, golden patches surrounding ocelli; head width 1.91-2.75 (2.28±0.21, 18), head width pronotum width ratio 0.89-1.0 (0.94±0.03, 18). Ocelli: three; black with white inserts; laterals larger than medial. Antennae: pedicel, scape and flagellum golden brown; more than twice length of head. Eyes: upper lobes of male orange-brown, lower lobes black; eyes of female black. Mouthparts. Labrum and clypeus ( Fig. 14 ): lateral margins of clypeus very slightly diverging towards anterior; lateral margins of labrum slightly wider than clypeus, rounded; anterior margin of labrum straight with no apparent antero-median emargination; labrum length 0.40–0.63 (0.51±0.07, 20); labrum width 0.95–1.33 (1.11±0.12, 20); width length ratio 2.00–2.45 (2.20±0.15, 20); five elongated denticles present on anterior margin; denticle width as proportion of labrum width 0.25–0.53 (0.37±0.08, 20); frontal setae arranged as a broad band; secondary hair fringe clearly separated from broad band. Mandibles ( Figs 15 & 16 ): outer margins slightly curved; small indentation and sparse long spine-like setae at midpoint on outer lateral margins, shorter setae along margin between midpoint and base; dark coloured nodule at midpoint on inner lateral margins; row of setae in an inverted L-shape on lower mandible body ventral surface. Left mandible ( Fig. 15 ): two incisors, each with three apical teeth, outer incisor inconspicuously serrated on inner lateral margins, prostheca stout, prosthecal tuft slender with hairs on lateral margins only. Right mandible ( Fig. 16 ): spine-like setae on inner lateral margins, two incisors, outer incisor with three apical teeth, inconspicuously serrated on inner lateral margins, inner incisor with two apical teeth, prostheca slender, prosthecal tuft slender with hairs on lateral margins only Maxillae ( Fig. 17 ): galea-lacinae as wide as long, width length ratio 0.85–1.17 (1.03±0.10, 10); subapical row of 16–23 (12) pectinate setae; palpi three segmented, terminal palp with single spine apically; palp length ratios 1.00: 0.69: 0.67 (0.37±0.04, 10). Hypopharynx ( Fig. 18 ): well developed lateral processes; anterior margin of lingua deeply cleft, lined with short setae; superlingua with thick tufts of setae on anterior margins, lateral margins slightly angular. Labium ( Fig. 19 ): glossae not turned under ventrally and lying in same plane as paraglossae, series of blunted spines apically; palpi three segmented, first segment with sharp spines, terminal segment with row of triangular spines almost circling apex and stout spines on dorsal surface; palp length ratios 1.00: 0.83: 0.50 (0.53±0.05, 10); submentum with spines on lateral margins. Thorax: tan brown with darker brown markings. Legs: yellow to tan brown, femora usually darker at apices; fore femora occasionally with white patch proximally but this is not always apparent. Foreleg ( Fig. 20 ): total length 4.96–7.33 (5.93±0.73, 15); fore femora length 2.25–3.13 (2.56±0.28, 15); fore femora width 0.63–0.90 (0.77±0.08, 15); fore femora length width ratio 2.88–4.05 (3.32±0.30, 15); fore tibiae with numerous short ventral spines, fore tarsi with 6–10 (10) ventral spines; tarsal claws ( Fig. 21 ) with 11–15 (10) ventral teeth, progressively larger apically, apical tooth inconspicuously serrated; leg length ratios, foreleg 1.00: 0.93: 0.36 (2.56±0.28, 15), mid leg 1.00: 0.87: 0.31 (2.52±0.29, 15), hind leg 1.00: 0.89: 0.27 (2.96±0.32, 15). Abdomen: tan brown with darker brown markings, golden markings medially on each segment, pattern variable; posterolateral spines present on segments two to nine, progressively larger posteriorly. Gills ( Fig. 22 ): membrane clear, light grey colour; present on segments one to seven, progressively smaller posteriorly; double, upper and lower lamellae equally developed; each gill tapered to a thin point apically; main and lateral tracheae present, strongly developed. Caudal filaments: three; golden brown; terminal filament longer than cerci.

Remarks. The larval stage of this species has been recognised previously and referred to as “Genus Y sp. 2” and Atalophlebioides “sp. 2” in various unpublished keys from the Museum of Victoria. In the preliminary guide to Australian Leptophlebiid nymphs it carried the name Nousia sp. AV3 ( Dean 1999).

Deanophlebia kala can be distinguished from other species in the genus by the following combination of characters. In the imago: (i) average male forewing length more than 11mm; (ii) male forewing hindwing ratio larger, forewing 4.7 times larger on average than hindwing; (iii) average male foreleg lengths longer than 10mm; (iv) abdomen with golden markings medially on each segment forming a broken stripe midline, two golden coloured maculae flanking each midline marking; (v) penis lateral margins more bulbous medially.

In the mature nymph: (i) denticles on labrum extend approximately two-fifths the total width of the labrum; (ii) average total foreleg length more than 5mm; (iii) average total foreleg femora length more than 2.5mm; (iv) abdomen with golden markings medially on each segment forming a broken stripe midline, two golden coloured maculae flanking each midline marking; (iv) gills with membrane clear.

Other specimens examined. NEW SOUTH WALES. Lake Albina outlet stream ( Lady Northcotes Creek ), 9 Feb 1985, 147 o 16′ 36 o 26′, 1920m, 3 ♂ N, 2 ♀ N, 1 ♀ S ( DC) . Lake Albina inflowing stream, Mt Kosciusko , 9 Feb 1985, 148 o 16′30′′ 36 o 26′59′′, 1920m, 1 ♀ N. ( JD) . Lake Albina , 20 Apr 1979, 147 o 16′ 36 o 26′, 1950m, 1 ♀ N ( IC) . Lake Cootapatomba inflowing stream, Mt Kosciusko , 24 Jan1984, 148 o 15′59′′ 36 o 28′01′′, 2160m, 1 ♂ N, 13 ♀ N ( JD) . Ramshead Creek , upstream Kosciusko Walking Track, 27 Jan 1984, 148 o 16′ 36 o 29′, 1940m, 1 ♀ N, ( JD) . South Ramshead , 25 Mar 1982, 148 o 14′ 36 o 31′, 1950m, 1 ♂ I ( RG) . Unnamed tributary, small creek crossing track, Upper Snowy Valley , Ramshead, 9 Feb 1985, 148 o 16′ 36 o 28′, 1940m, 1 ♂ N ( JD) . Unnamed creek, south slope of North Ramshead , 27 Jan 1984, 148 o 16′ 36 o 28′, 1940m, 3 ♂ N, 2 ♀ N ( JD) . VICTORIA. Albert River, Albert River Rd , 31 May 1984, 146 o 23′26′′ 38 o 30′22′′, 330m, 3 ♂ N, 4 ♀ N. ( DC) . Badger Creek , downstream weir: 18 Jan 1980, 145 o 34′ 37 o 41′, 280m, 1 ♂ I (reared) ( JD) ; 20 Mar 1980, same locality data, 1 ♀ NS (reared) ( JD) . Badger Creek , upstream weir, Badger Weir Park, 5km SE of Healesville, 18 Jan 1980, 145 o 34′04′′ 37 o 41′17′′, 300m, 5 ♀ N ( JD) . Buckland River, Mt Buffalo Rd , 7 Nov 1996, 146 o 53′32′′ 36 o 41′52′′, 280m, 1 ♀ N ( KJF) . Bull Creek, Bull Creek Rd, 30 Nov 1996, 145 o 35′39′′ 37 o 28′31′′, 360m, 1 ♀ NS (reared), 1 ♀ N ( KJF) . Bull Creek tributary, W of Yea Rock, 30 Nov 1996, 145 o 35′14′′ 37 o 27′07′′, 450m, 1 ♀ N ( KJF) . Buller Ck, Mirimbah, 2 Nov 1981, 146 o 24′ 37 o 06′, 670m, 4 ♂ N, 7 ♀ N ( JD) . Cement Creek, Mount Donna Buang Rd : 8 Apr 1976, 145 o 42′20′′ 37 o 42′48′′, 670m, 1 ♀ N, 1 ♀ NS (reared), 1 ♀ NSI (reared) ( JD) ; 23 Apr 1976, 1 ♀ NSI (reared) ( JD) ; 1 Dec 1976, 1 ♂ I ( JD) ; 14 Apr 1977, 1 ♂ N, 1 ♂ S, 1 ♀ I, 1 ♀ S; 12 Jan 1978, 4 ♀ N, 1 ♂ I ( JD) ; 6 Oct 1978, 1 ♂ NI (reared) ( JD) ; 7 Dec 1979, 1 ♂ NI ( JD) ; 3 Mar 1980, 3 ♂ I, 3 ♀ I, 3 ♀ S ( JD) ; 25 Sept 1996, 1 ♀ NS ( KJF) ; 5 Mar 1998, 1 ♀ N ( KJF) ; 23 Jan 1999, 1 ♀ NSI (reared), 1 ♀ NS ( KJF) ; 26 Feb 2000, 1 ♂ NSI (reared), 1 ♂ N ( KJF) . Charity Creek tributary, Mt Baw Baw Tourist Rd , 16 Jan 1999, 146 o 15′15′′ 37 o 50′47′′, 1180m, 1 ♂ NSI, 1 ♀ NSI, 1 ♀ NI, 1 ♂ NS, 1 ♂ N ( KJF) . Clearwater Creek, Bogong High Plains Rd, 10 Nov 1996, 147 o 21′14′′ 37 o 57′10′′, 1410m, 1 ♀ N ( KJF) . Clematis Creek , Monbulk Rd, 15 Oct 1997, 145 o 21′22′′ 37 o 53′57′′, 290m, 2 ♀ NSI (reared), 1 ♂ NS (reared) ( KJF) . Coranderrk Creek , Badger Weir Park, 06 Apr 1997, 145 o 34′16′′ 37 o 41′07′′, 240m, 2 ♂ N, 1♀ N ( KJF) . Donnelly Creek , Donnelly Weir Rd, 6 Apr 1997, 145 o 32′03′′ 37 o 37′40′′, 140m, 1 ♀ NS (reared), 1 ♂ N ( KJF) . Errinundra River , 1.5km upstream Combienbar Rd, 21 May 1981, 148 o 55′ 37 o 27′, 120m, 2 ♂ N ( JD) . Falls Creek , Telephone Box Junction, Mt Stirling Rd, 2 Nov 1981, 146 o 27′43′′ 37 o 06′52′′, 1220m, 1 ♀ N ( JD) . Four Mile Creek , Horners Rd, Warburton, 25 Apr 1997, 145 o 41′56′′ 37 o 45′33′′, 185m, 1 ♀ NS (reared) ( KJF) . German Creek , Bright-Tawonga Rd, 9 Nov 1996, 440m, 147 o 03′25′′ 36 o 43′58′′, 440m, 1 ♂ N ( KJF) . Grace Burn upstream weir, 27 Apr 1978, 145 o 35′02′′ 37 o 39′26′′, 250m, 1 ♀ NS (reared) ( JD) . Grace Burn, Wallace Parade, Healesville , 6 Apr 1997, 145 o 32′18′′ 37 o 39′16′′, 110m, 1♂ N ( KJF) . Hope Creek , Tanjil Bren Rd, 12 Oct 1996, 146 o 14′03′′ 37 o 51′55′′, 550m, 1 ♀ NS (reared), 2 ♀ N ( KJF) . Icy Creek , Tooronga Rd, 11 Oct 1996, 146 o 00′50′′ 37 o 48′09′′, 530m, 2 ♀ N ( KJF) . Keppel Creek , upper site: 17 Nov 1994, 145 o 46′ 37 o 27′, 800m, 8 ♀ N ( SP) ; 10 Oct 1994, 1 ♀ NS (reared) (SP); 19 Oct 1994, 1 ♂ NS (reared) (SP); 05 Dec 1994, 1 ♀ NS (reared) (SP); 13 Mar 1995, 1 ♂ S (SP); 14 Mar 1995, 1♂ NSI (reared), 2 ♀ NSI (reared), 1 ♂ NS (reared), 1 ♀ NS (reared) (SP); 15 Mar 1995, 1 ♂ NS (reared), 1♀ NS (reared) (SP); 21 Mar 1995, 1 ♀ I (SP); 27 Mar 1995, 1 ♂ NS (reared) (SP); 31 Mar 1995, 1 ♂ NSI (reared) (SP); 10 Apr 1995, 1 ♀ NSI (reared) (SP). King Parrot Creek, Wallaby Creek Rd, 16 Aug 1976, 45 o 14′56′′ 37 o 25′50′′, 350m, 2 ♀ S ( JD) , 1 ♂ I ( JD) . King Parrot Creek , 4WD track off Whittlesea-Yea Rd, Kinglake West, 7 Oct 1996, 145 o 15′44′′ 37 o 25′20′′, 340m, 1 ♂ N ( KJF) . Lake Elizabeth , inlet stream, Otways, 22 May 1988, 143 o 45′36′′ 38 o 33′13′′, 210m, 2 ♂ N ( DC) . Loch River , Noojee, 2 May 1993, 145 o 59′ 37 o 53′, 240m, 1 ♂ NSI ( SP) . Martins Creek No. 2, Bonang Rd, 0.5km S of Malinns , 13 Feb 1999, 148 o 36′ 37 o 25′, 260m, 1 ♀ N ( KJF) . Mt Baw Baw : 19 Nov 1975, 146 o 16′ 37 o 50′, 1460m, 11 ♂ I ( IC) ; 29 Nov 1975, 2 ♂ I, 3 ♂ S, 1 ♀ S ( IC) . Mt Buller , 28 Mar 1995, 146 o 25′ 37 o 08′, 1680m, 3 ♂ I, 2 ♀ I, 1 ♀ S, 3 ♀ N ( IC) . Myrtle Gully, Sylvia Creek Rd, Toolangi State Forest , 29 Nov 1996, 145 o 31′44′′ 37 o 31′44′′, 620m, 1 ♂ N, 3 ♀ N ( KJF) . North Cascade Creek, Thomson Valley Rd , 13 Jan 1991, 146 o 20′19′′ 37 o 48′22′′, 920m, 1 ♀ N ( JD) . O’Shannassy River : 14 Nov 1975, 145 o 49′ 37 o 36′, 580m, 1 ♂ N ( JD) ; 4 Jan 1980, 1 ♂ N, 1 ♀ N ( JD) . Perrins Creek , Warwick Rd, 24 Mar 1997, 145 o 22′ 37 o 52′, 350m, 1 ♂ NSI (reared), 1 ♂ I, 1 ♀ NS ( KJF) . Pretty Valley Creek, Bogong High Plains Rd, 1.5km S of Bogong, 9 Nov 1996, 147 o 13′20′′, 36 o 49′10′′, 740m, 2 ♀ N ( KJF) . Razor Creek , Mt Stirling Rd, 30 Mar 1989, 146 o 27′19′′ 37 o 06′52′′, 1160m, 1 ♀ N ( AN) . Rum Creek, Thomson Valley , 17 Apr 1985, 146 o 22′49′′ 37 o 52′45′′, 640m, 1 ♂ N, 5 ♀ N, ( JD) . Running Creek , Warburton-Jamieson Rd, 31 Oct 1981, 145 o 58′ 37 o 25′, 750m, 1 ♂ N, 2 ♀ N ( JD) . Running Jump Creek tributary, Mt Buffalo Rd , 7 Nov 1996, 146 o 46′26′′ 36 o 46′15′′, 1500m, 1 ♀ N ( KJF) . Sassafras Creek, Perrins Creek Rd, 15 Oct 1997, 145 o 22′18′′ 37 o 52′46′′, 370m, 1 ♀ NSI (reared) ( KJF) . Snobs Creek tributary, Snobs Creek Rd, 31 Oct 1981, 145 o 54′38′′ 37 o 21′15′′, 760m, 10 ♀ N ( JD) , 4 ♂ N ( JD) . Starvation Creek , upstream weir, 28 Oct 1977, 145 o 50′53′′ 37 o 45′39′′, 340m, 1 ♂ S ( JD) . Talbot Creek: downstream Thomson River Reservoir , 10 Aug 1983, 147 o 22′20′′ 37 o 50′52′′, 700m, 1 ♂ N, 7 ♀ N ( DC) ; 9 Apr 1986, 3 ♂ N, 1 ♀ N ( DC) . Tanjil River east branch tributary, Mt Baw Baw Tourist Rd , 11 Oct 1996, 146 o 11′29′′ 37 o 49′58′′, 500m, 1 ♂ NS (reared) ( KJF) . Tarra River, Tarra-Bulga National Park , 9 Jan 1984, 146 o 32′13′′ 38 o 26′57′′, 340m, 1 ♀ N ( JD) . Tarra River , 20m upstream Tarra Falls, 9 Dec 1984, 146 o 32′20′′ 38 o 27′17′′, 320m, 4 ♂ N, 6 ♀ N ( JD) . Tarra River, Tarra-Bulga National Park , 16 Feb 1999, 146 o 32′15′′ 38 o 27′00′′, 340m, 1 ♀ NSI (reared), 4 ♂ N, 2 ♀ N ( KJF) . Traralgon Creek , Guntzler Bridge, 29 May 1984, 146 o 31′ 38 o 20′, 110m, 3 ♂ N, 2 ♀ N ( DC) .

Figures

FIGURE 13. Deanophlebia kala. Female nymph, scale bar 1mm.

FIGURES 1–7. Deanophlebia kala. Male imago: Fig.1. SEM of foreleg tarsal claw, scale bar 50µm; Fig. 2. Forewing showing relative size of hindwing, scale bar 1mm; Fig. 3. Hindwing, scale bar 500 µm; Fig. 4. Dorsal view of abdomen, scale bar 1mm; Fig. 5. Genitalia, dorsal view, scale bar 200 µm; Fig. 6. Genitalia, ventral view, scale bar 200 µm. Fig. 7. SEM close up of right lobe of penes showing apical process, scale bar 50 µm.

FIGURES 8–12. Deanophlebia kala. Female imago: Fig. 8. Forewing showing relative size of hindwing, scale bar 1mm; Fig. 9. Hindwing, scale bar 500 µm; Fig. 10. Ventral view of abdomen, sternites seven to nine, scale bar 500µm; Fig. 11. SEM of egg, scale bar 50 µm; Fig. 12. SEM detail of egg chorion, scale bar 10µm.

FIGURES 14–22. Deanophlebia kala. Nymph: Fig. 14. Labrum and clypeus, scale bar 200µm; Fig. 15. Left mandible, scale bar 200µm; Fig. 16. Right mandible, scale bar 200µm; Fig. 17. Maxilla, scale bar 200µm; Fig. 18. Hypopharynx, scale bar 200µm; Fig. 19. Labium, scale bar 200µm; Fig. 20. Foreleg, scale bar 500µm; Fig. 21. Foreleg tarsal claw, 100µm; Fig. 22. Gills, scale bar 100µm.