Iheringia Three new species of Leptohyphes Eaton ( Ephemeroptera : Leptohyphidae ) from Colombia

Three new species of Leptohyphes Eaton, 1882 (Ephemerotera: Leptohyphidae) are described, diagnosed and illustrated from nymphs collected in Colombia. Geographical ranges and ecological notes relating important environmental variables are given for each species. Leptohyphes andina sp. nov. is characterized by: forefemur length / maximum width 1.8-1.9, fore and hind margin of middle and hind femora with strong spine-like setae on elevated sockets, femoral surface covered by small pale spots, apex of femora with anterior margin roundly projected distally, tarsal claw only with 3-4 marginal denticles, pronotum with triangular lateral projection, hind wing pads present in female, gill formula 3/5/5/5/2, gill V with ventral extension on dorsal lamella. Leptohyphes quimbaya sp. nov. presents the following characters: body covered by hair-like setae, with paired short and blunt tubercles on abdominal terga V-IX, femora wide (forefemur length/ maximum width 1.6), fore and hind margin of femora with tiny spine-like setae, tarsal claws with 2-3 marginal denticles, pronotum with triangular (but apically blunt) lateral projection, hind wing pads present in females, gill formula 3/7/7/5/2, gill V without ventral extension on dorsal lamella. Leptohyphes calarca sp. nov. is distinguished by: pilose body, slender femora (forefemur length / maximum width 2.3), foremargin of middle and hind femora without setae, tarsal claws with 5 marginal denticles and 1 submarginal subapical denticle, pronotum strongly projected laterally, fore wingbuds blackish, hind wing pads present in females, gill formula 3/4/4/3/1, gill V with ventral extension on dorsal lamella.

The aim of the present paper is to describe and illustrate the nymphal stage of three new species collected in the Andean region of Colombia.

MATERIAL AND METHODS
Nymphs were collected with diff erent aquatic nets (surber sampler, kicknet, D-frame net, hand nets) and fi xed and preserved in 96% ethanol.Mouthparts and legs were dissected under 10x magnifi cation and mounted on microscope slides, using Euparal® or Canada Balsam.The character "gill formula" was used in descriptions of species in the form of a numeric series (e.g., 3/11/9/9/6), where the first number represents the number of gill lamellae present in each gill on the first abdominal segment, second number represents the number of lamellae present in each gill of the second abdominal segment and so on.The tarsal claw denticulation is also represented as a formula (e.g., 2-3 + 0) indicating first the number of marginal denticles on the basal half of the claw (in the example varying from 2 to 3) plus the number of apical submarginal denticles (in this case, absent), following Molineri (2003).The presence of hind wings (or wing pads) is only mentioned in females because they are always present in the males of the genus (Molineri, 2003).
Photographs were taken using a stereomicroscope with a digital camera.Line drawings were done using a camera lucida attached to a microscope.Maps were made with Geographic Coordinate System: GCS Bogotá, Datum: D Bogotá and the software ESRI® ArcMap 10.0 licence ArcInfo.
Etymology.The specific epithet refers to the Andes.Distribution (Fig. 28).Colombian Andes (Central and Western Cordillera) in the departments of Quindio, Risaralda and North of Valle del Cauca, from 750 to 1648 masl.
Discussion.The unique combination of characters distinguishing L. andina from other species in the genus is listed in the diagnosis.Some of these characters are shared by other species, as follows: pronotum with triangular lateral projection (also present in L. quimbaya), femora with pale spots (present in L. liniti Wang et al., 1998, L. petersi Allen, 1967, L. murdocki Allen, 1967, L. alleni Brusca, 1971), femora with spine-like setae on fore and hind margins (also present in many species except those mentioned above for the pale spots).The femora (middle and hind) with inner apical corner more projected than the outer is a unique characteristic, not reported before in the genus.
Ecological notes.Nymphs were found mainly in small streams from the middle basin of La Vieja river (departments of Valle del Cauca and Quindío).The area is a human driven ecosystem with coffee and banana plantations, and cattle rising as the main activities.The marginal areas of the streams presented some kind of plant cover, mainly herbs, bushes, and bamboo (Guadua angustifolia Kunth, 1822) or relatively well-preserved secondary forests.These streams varied from 4 to 20 cm in depth, 46 to 606 cm in channel width, and 0.5 to 31 l/s of discharge.Bed substrate is formed by gravel (11%), stones (26.6%) and sand/slime (62.8%).Mean water temperature was 21°C and did not present severe organic loads.Mean ICA-FSN quality index was 60.6 corresponding to good environmental quality (Ott, 1981).Other variables are: mean dissolved oxygen (5.2 mg/l), low biochemical oxygen demand (0.69 mg/l), and variable pH (5.8 to 8.4) indicating an acid-basic stable buffer system (Chará et al., 2009).
Description.Nymph.Length of female (mm, not fully mature): body, 7.0; hind femur, 1.7; caudal filaments, 5.5 (cercus)-6.8 (terminal filament).General coloration yellowish brown, with gray markings (Figs 2, 6-7).Head yellowish with a blackish transverse band between eyes along the epicranial suture; occiput with small gray marks as in Fig. 6, posterior margin with a thin blackish line.Antennae whitish.Thorax: pronotum with large rounded anterolateral projection and pronounced constriction posteriorly, with gray markings as in figure (Fig. 6).Mesonotum with blunt anterolateral projections, and with gray markings as in Fig. 2; fore wingbuds black.Hind wing pad present in female.Thoracic sterna paler than terga, shaded with gray medially.Legs yellowish, shaded with gray on coxae, femora (except mediolongitudinal pale line), and base of tibia; legs almost completely covered by dark thick and short setae (Fig. 7), except for bare mediolongitudinal dorsal line on femora.Foreleg (Fig. 18): femur slender, becoming wider apically, ratio length/maximum width 2.3; transversal row at 0.4 from base to apex, with 25 stout spatulate spine-like setae; fore margin with spine-like setae, hind margin with spine-like setae (on elevated sockets) distally to transversal row; small inner projection on apex of femur.Tibia subequal in length to femur with spatulate setae along inner margin.Tarsus 0.4 the length of tibia with row of setae in inner margin; tarsal claw with 5 +1 denticles (Fig. 17).Middle and hind legs (Fig. 16) similar except for the size; hind femur becaming wider apically, ratio length/maximum width 2.4, with double row of spine-like setae on fore margin, hind margin with 39 spinelike setae on elevated sockets; projections on inner apex of femur small.Tibia wide, 1.2 the length of femur, with dorsal ridge, inner and outer margin row of large spatulate setae (Fig. 16).Tarsus 0.3 the length of femur, inner margin with setae, tarsal claw as in foreleg.Abdomen.Terga brownish, shaded uniformly with gray; few spatulate setae present on lateral areas of segments III-VII.Lateral margins of segments III-VI expanded forming flanges; relatively short posterolateral spines on segments VII-IX.Abdominal sterna shaded slightly with gray.Sternum IX with distal V-shaped indentation (female).Gills: operculate gill on segment II whitish, shaded with brown on basal half, ventrally with curved "basal spine" and two lamellae (Fig. 19); other gills whitish (Fig. 20).Gill formula 3/4/4/3/1.Caudal filaments yellowish with few short and blunt brownish spines.Adults.Unknown.The wing membrane is surely tinged with gray or black because of the pigments shown in the nymphal wing buds.
Etymology.Calarcá is the name of the municipality where the nymphs were collected; the name refers to the Iheringia, Série Zoologia, 106: e2016024 legendary Cacique Calarcá, from an ethnic group known as Pijaos who occupied the Colombian central Andean region.
Discussion.The large size and pilose legs resembles L. quimbaya, but L. calarca is even larger and hair-like setae on the head, thorax and abdomen are lacking.Other important differences include the larger and rounded anterolateral projection on pronotum, the blackish wing buds and the presence of subapical submarginal denticle in tarsal claws of L. calarca.
Ecological notes.Known locality records of this species are from La Sonadora Natural Reserve, on the eastern slope of the Central Cordillera, Quindío (Calarcá and Córdoba municipalities).They were found between 2800 and 3100 m.The region present 2000-4000 mm of annual rainfall and temperatures ranging 9-16°C (Baquero et al., 2010), with constant winds and fog.The area corresponds to Montane Humid Forest in Holdridge's (1987) classification.Streams are small, low-order reaches located in the high microbasin of La Sonadora and draining to the Verde river basin.They flow through a scarped terrain, with high slopes and falls of 3 to 15 m high, with well-preserved dense marginal vegetation cover.The streams presented a depth of 18-35 cm, channel with of 130-270 cm, and velocity of 0.15-0.50m/s.Bed substrate is muddy with high levels of leaves and other decomposing organic material, but sandy and stony substrate is present also in some creeks, but in a lower proportion.
Description.Mature nymph.Length of female (mm): body, 7.1-9.2;hind femur, 1.9-2.0;caudal filaments, 7.5.General coloration light brown, body and legs dorsally covered with whitish hair-like setae, venter glabrous.Head with black transverse band from eye to eye as in Fig. 21; occiput with finely marked gray net-pattern (Fig. 21).Thorax: nota brownish with gray markings.Pronotum subrectangular with small lateral subtriangular projection, and an oblique indentation near posterior margin (sp and oi in Fig. 22).Mesonotum with anterolateral corners not projected (Fig. 22).Hind wing pads present in female.Legs yellowish shaded dorsally with gray except mediolongitudinal narrow band on all femora (Fig. 9).Foreleg (Fig. 24): femur wide, ratio length/maximum width 1.6; transversal row of setae at 1/2 from base; fore margin with setae, hind margin with setae (on elevated sockets); apex of femur projected on inner and outer margins (Fig. 24).Tibia subequal in length to femur with row of setae on inner margin; tarsus 0.4 the length of tibia with row of setae on inner margin; tarsal claw with two to three marginal denticles and none subapical submarginal denticle (Fig. 25).Middle and hind legs (Fig. 23) similar, except hind leg larger; hind femur ratio length / maximum width = 2; fore margin with 3-4 rows of short blunt setae, hind margin with 70 stout spine-like setae on elevated sockets (the sockets and the spines are very small), dorsal surface with many small scattered spine-like setae; distal projections on apex of femur more pronounced than foreleg.Tibia 1.1 the length of femur, with dorsal ridge, inner and outer margins with short weak spine-like setae.Tarsus 0.3 the length of tibia, inner margin with setae, tarsal claw as in foreleg.Abdomen (Figs 26,27) brownish shaded widely with gray, ventrally paler.Terga (Figs 9,26) covered by whitish hair-like setae, mainly along hind margin; short blunt spine-like setae scattered on terga, mainly on submedian tubercles; segments III-VI with lateral flanges, small and poorly developed posterolateral projections on VII-IX; sternum IX with distomedial notch.Gills: operculate gill completely shaded with gray, except margins; other gills whitish shaded slightly with gray.Gill formula 3/7/7/5/2; gill V without ventral extension.Caudal filaments with scattered setae, and whorl of spines at joinings.
Variation: early instars of L. quimbaya only show groups of small setae on the area where abdominal tubercles will develop, and their almost circular forefemur does not present the mediolongitudinal row of spine-like setae distally.
Adults.Unknown.Etymology.Quimbaya refers to an ethnic group that inhabited the valley of the Cauca river, between the Central and Western Cordilleras in Colombia, now the departments of Caldas, Quindío and Risaralda.
Discussion.The uncommon large body size together with the many tiny femoral spine-like setae, the setose body, very wide femora and paired abdominal tubercles readily distinguish this species from any other in the genus.Leptohyphes quimbaya is similar to L. pilosus Allen & Brusca, 1973, mainly in the body pilosity, but the last species presents fewer and larger spine-like setae on slender femora.Abdominal tubercles are present in few other species of the genus, but only in L. nebulosus Nascimento et al., 2014 they are paired, nevertheless the later presents spine-like setae on very elevated sockets on femora, and is restricted to Brazilian coastal Atlantic forest (Nascimento et al., 2014).
Ecological notes.The species was collected in several creeks on the eastern slopes of the Western Cordillera in Colombia (Risaralda department).The landscape spans elevations from 1300 to 2000 m, where sun-grown coffee is dominant (WCS, 2013) and corresponds to the transition zone between the Lower Montane Wet Forest and Premontane Moist Forest (Espinal 1977).Mean annual rainfall is 2500 mm and mean annual temperature is 18°C.The landscape is a mosaic of vegetation types dominated by patches of native vegetation that differ in size (located at the top of the mountains and in glens), sun-grown coffee plantations, annual crops, stover and pasture (WCS, 2013).The Verdum Municipal Natural Park in this study area is located in the Iheringia, Série Zoologia, 106: e2016024 buffer zone of the Tatamá National Natural Park and it is important in environmental terms as a reservoir of Andean forest.