New species and records of Zebragryllus Desutter-Grandcolas & Cadena-Castañeda, 2014 (

. Zebragryllus Desutter-Grandcolas & Cadena-Castañeda, 2014 is a Neotropical genus of field crickets that inhabits the leaf litter of the Amazon rainforest. The genus has six species and is characterized by the distinct ‘zebra’ pattern displayed by some of them. The species are recorded in French Guiana, Peru, and Colombia. Here, we describe two new species of the genus from the Brazilian Amazon rainforest, the first record of the genus for Brazil. We expand the records of Z. guianensis Desutter-Grandcolas, 2014 and Z. nouragui Desutter-Grandcolas, 2014 to Brazil. We also describe but do not name two females corresponding to two new species, provide an up-to-date key for species of Zebragryllus , and discuss the potential distribution for the genus.


Introduction
Despite its high diversity, the subfamily Gryllinae, Laicharting, 1781 is mainly found in regions of Africa and Eurasia, and the cosmopolitan tribe Gryllini, Laicharting, 1781 is the only group to occur in the Neotropics (Cigliano et al. 2023).However, no representative of this subfamily was ever reported to occur in South American caves.
In this work, we describe two new species of Zebragryllus, describe but do not name two additional females comprising two new species, and expand the records of Z. guianensis Desutter-Grandcolas, 2014 andZ. nouragui Desutter-Grandcolas, 2014 to Brazil.In addition, we provide an up-to-date identification key for species of the genus and discuss its potential distribution.

Material and methods
The genitalia were removed and treated with a warm 10% aqueous solution of potassium hydroxide (KOH) for approximately 10-15 minutes to digest and remove muscles and fat.Afterward, they were stored in an 80% ethanol solution, and both body and genitalia morphologies were analyzed under a Leica M205 A stereo microscope equipped with a Leica DFC 450 camera.
The venation terminology followed Desutter-Grandcolas et al. (2017).The nomenclature adopted for the phallic complex followed Desutter (1987), modified in Desutter-Grandcolas (2003) and is defined in the section Abbreviations for measurements and morphological terms.
Measurements of body parts were made using a digital pachymeter and were defined as follows: total length (TL) as the distance between the top of the fastigium of vertex and the apex of the supra-anal plate; length of pronotum (LP) as the length of a mid-dorsal line between the anterior and posterior margins of the pronotal disk; width of pronotum (WP) as the maximum distance between the midportion of pronotum lateral lobes; tegmina (Tg) as the length of a dorsal line between the base and the apex of tegmina; hind femur (HF) as the distance between the base of the hind femur and the apex of genicular lobes; hind tibia (HT) as the distance between the base and the apex of the hind tibia; and ovipositor (Ov) as the distance from the apex of the subgenital plate and the apex of the ovipositor.
All the type material is deposited in the Entomological Collection of the Museu Paraense Emílio Goeldi (MPEG), in Brazil.Additional non-type specimens were distributed between two institutions, MPEG and Coleção de Invertebrados Subterrâneos de Lavras (ISLA), in the Universidade Federal de Lavras, also in Brazil.

Potential distribution
The study of species' potential distribution can be facilitated through maps that utilize Minimum Convex Polygons (MCPs) (Mace et al. 1992).This method involves creating minimum convex polygons, which are the smallest polygons where no internal angle exceeds 180° and that encompass all occurrence points (IUCN 2021).For the genus Zebragryllus, in which most species have few occurrence points (Cigliano et al. 2023), we developed an MCP for the genus as a model by using the coordinates of the published records of species, available in Desutter-Grandcolas et al. (2014) and Cigliano et al. (2023), and new records from this study.The maps were produced using the QGIS software (https://QGIS.org).B, 4A-B).Tympana reduced or absent (Fig. 2A-F).All femora entirely ochre, without the 'zebra' pattern (Fig. 2A-C, G-K).Males dorsally with head, pronotum, tegmina, and first abdominal tergite black (Fig. 1A-E); without stridulatory apparatus (Fig. 1E).Comparatively, all the other known species of Zebragryllus are easily distinguished for having stridulatory apparatus on the male tegmina.Male genitalia: pseudepiphallic median lophi elongated, conspicuously produced hindand upwards, very narrow, and apically acute, ventrally with a sharp keel.Pseudepiphallic lateral lophi acute.Pseudepiphallic paramere surpassing the lateral lophi; in ventral view, slightly arched.Ectophallic fold very thin, not reaching the level of the pseudepiphallic paramere apex (Fig. 3A-D).Female with the black coloration extending till the second abdominal tergite (Fig. 4A-D); remaining tergites dark brown; ovipositor straight and almost as long as the hind femora (Fig. 4A, H-I).Female genitalia: copulatory papilla with posterior margin of dorsal surface widely concave, and the anterior margin with medial process emarginated, flanking the spermatheca duct (Fig. 4L); ventral surface acute anteriorly (Fig. 4M); in lateral view, posterior portion taller than anterior one (Fig. 4N).

Etymology
The specific epithet "aphonus" refers to the absence of a stridulatory apparatus, which makes the cricket incapable of producing sound.
Paratypes BRAZIL • 1 ♀; same collection data as for holotype; except for the left hindleg, all the remaining legs have fallen but are stored in the same specimen vial, the subgenital plate, the seventh abdominal sternite, and the copulatory papilla were removed but kept in a microvial with the specimen; MPEG.HEX 05050608 • 1 ♂; same collection data as for preceding; S11C-0066 cave; 6°23′53.5″S, 50°22′58.1″W; 9 Oct. 2022; except for the left foreleg and right hindleg, all the remaining legs have fallen but are stored in the same specimen vial; MPEG.HEX 05050609.
Head.In frontal view (Fig. 1C), semicircular, almost as wide as high, with eyes slightly produced laterally; ocelli circular and almost aligned to eyes dorsal margin -median ocellus slightly lower.Antennal pits located low on frons, near epistomal suture, and almost on same level as eyes' most ventral border (these last also close to subgenal suture).Clypeus dorsal area notably separated from ventral area by clypeal suture.Ventral area conspicuously constricted laterally, almost half as wide as dorsal area, with median white stripe and transverse dark line on sides.Labrum circular, with pair of incomplete transverse mediolateral sutures.First two segments of maxillary palpi subequal in length; third and fourth as long as first and second together; fifth longer than any other segment, ventrally truncated on mediodistal area (Fig. 1G).In lateral view (Fig. 1D), head convex, with frons slightly tumescent; eyes ovoid, higher than wide.In dorsal view (Fig. 1B), eyes protruding; fastigium subtriangular and apically convex.Scapus wide, almost as wide as half fastigium apex.tHorax.Pronotum wider than long, bearing bristles on margins, with both anterior and posterior dorsal margins slightly concave (Fig. 1B).Lateral lobes almost squared, with anterior and posterior margins almost straight and ventral margin obliquely straight (Fig. 1A, D).Prosternum reduced, triangular, and continuous with conjoint cervical sclerites.Mesosternum sub-squared, with posterior margin slightly bilobed.Metasternum hexagonal, with posterior margin slightly incised (Fig. 1F) legs.All femora covered by fine pubescence and some longer bristles (Fig. 2A-C, G-K).Fore and mid tibia with two apical spurs on outer side (Fig. 2A-C, H) and one on inner side (Fig. 2G, I).Tympana reduced (Fig. 2A-B, D-E) or absent (Fig. 2C, F) (different development levels can be found in same  specimen).Hind tibia with 6-7 external and 5-6 internal dorsal spurs and three apical spurs on each side (Fig. 2J-K); externally, dorsal and ventral apical spurs equal in length, mid one twice longer; internally, dorsal and mid spurs same-lengthened, ventral one at least three times smaller (Fig. 2J-K).Fore and mid basitarsi covered by strong setae on ventral side (Fig. 2A-C, G-I, L).Hind basitarsi ventrally with setae, dorsally with two rows of spines, and apically with two spurs, outer one smaller (Fig. 2J-K).

Remarks
Zebragryllus aphonus Tavares, Oya & Cadena-Castañeda sp.nov.has quite a unique feature.It is the only known species of the genus in which the male has no stridulatory apparatus.The different development levels of the tympana are also singular to this species.These two characteristics may be adaptations to the caves in which they were found.Still, it is hard to ensure since no phylogenetic study was made and ecological data are lacking.The shape of the pseudepiphallic median lophi is similar to that of Z. nauta.However, it differs by the presence of conspicuous pseudepiphallic lateral lophi, which are very reduced and almost inconspicuous in Z. nauta.These two species may be related.
We believe this species is facultative to a cave-dwelling way of life or caviculous (sensu Desutter-Grandcolas et al. 1998), foraging on forest ground during the nights and passing the day inside the caves or cavities at ground level.We believe in this assumption since adult and immature stages were found inside caves as field work was carried out during the day.This species may also inhabit or take refuge in pores or voids on the soil, characteristic microhabitats of the environment where it was collected.The structure of the landscape is known as Banded Ferruginous Formations, which are iron formations composed of alternated bands of jaspilite and ore bodies rich in iron.In this area, extensive iron ore plateaus are formed, comprising superficial ferrugineous breccia, also known as canga formation.A metallophilic savannah covers the canga formations on the plateaus and mountains, while typical tropical vegetation of the Amazon rainforest covers the slopes.The highly porous canga formations result from the removal of silica and carbonates due to the tropical rains, which causes the dissolution and chemical change found in iron ore, forming several interconnected small spaces in the canga formations -the voids (Ferreira et al. 2018).

Etymology
The specific epithet is an homage to the indigenous community that lives in the type-locality area, the Mebêngôkre, which is the self-denomination of natives known in Brazil as the Kayapó people.The name is an indigenous word of the Macro-Jê linguistic trunk and is established as a noun in apposition.
Head.In frontal view, semicircular, slightly higher than wide, with eyes not laterally protruding; all three ocelli circular and almost at same level of eyes dorsal margin -median ocellus slightly lower.Antennal pits located near mid-level of eyes.Clypeus dorsal portion notably separated from ventral portion by clypeal suture.Ventral portion conspicuously constricted laterally, almost half as wide as dorsal portion, with median white stripe and transverse dark line on sides.Labrum circular, with pair of incomplete transverse mediolateral sutures.First two segments of maxillary palpi subequal in length; third and fourth each as long as first and second together; fifth longer than any other segment, with ventral margin slightly arched, dorsal margin straight, and apex rounded.Distal portion of each maxillary palpomere whitish (Fig. 6G).In lateral view, head notably convex, but frons not tumescent; eyes ovoid, and higher than wide (Fig. 6D).In dorsal view, eyes protruding, with trapezoid and apically truncated fastigium (Fig. 6B).Scapus wide, almost as wide as half of fastigium.tHorax.Pronotum wider than long, bearing bristles on margins and posterior area of lateral lobes, with both anterior and posterior dorsal margins slightly concave.Lateral lobes almost squared, with anterior and posterior margins straight and ventral margin obliquely straight.Prosternum reduced, triangular, and continuous with conjoint cervical sclerites.Mesosternum sub-squared, with posterior margin slightly bilobed.Metasternum hexagonal, with postero-lateral margins convex and posterior margin straight (Fig. 6F).
legs.All femora covered by fine pubescence and some longer bristles (Fig. 7A-B, D-E, G-H).Fore and mid tibia with two apical spurs on outer side (Fig. 7A, D) and one on inner side (Fig. 7B, E).Tympana large, opened on fore tibia outer surface (Fig. 7A, C).Hind tibia with six outer and five inner dorsal spurs, and three apical spurs on each side (Fig. 7G-H); externally, dorsal and ventral apical spurs equal in size, and mid one twice longer; internally, dorsal and mid spurs equal in size, and ventral one at least three times smaller (Fig. 7G-H).Fore and mid basitarsi covered by strong setae on ventral side (Fig. 7A-B, D-F).Hind basitarsi ventrally with setae, dorsally with two rows of spines, and apically with two spurs, outer one smaller (Fig. 7G-H).

Female
Unknown.

Nymph female
The coloration is very similar to the adult male but lighter (Fig. 9A-B).

Remarks
This species also has quite unique male genitalia, with very long pseudepiphallic median lophi.The only known species with elongated pseudepiphallic median lophi are Z.nauta and Z. aphonus Tavares, Oya & Cadena-Castañeda sp.nov., but Z. mebengokre Tavares, Oya & Cadena-Castañeda sp.nov.has the most extended and most robust pseudepiphallic median lophi of all species of Zebragryllus.
The locality where the species was collected is in a transition zone between two large biomes, the Cerrado-Amazonia transition, considered the world's largest ecotone (Torello-Raventos et al. 2013;Marques et al. 2020) (Fig. 16A-C).Marked seasonality and highly dynamic vegetation, comprising a mosaic of Cerrado, Amazon rainforest, and ecotonal forest vegetations, with high tree mortality and turnover levels, characterize this zone (Marimon et al. 2014;Morandi et al. 2015;Passos et al. 2018;Marques et al. 2020).

Diagnosis
This female is similar to that of Z. wittoto, Z. guianensis, and Z. nouragi by the 'zebra' pattern on the hind legs (Figs 10A-B, 11J-K).The fore and mid legs have only a few whitish areas on the base of the femora and dorsal surface of the tibia (Fig. 11E-F, H-I).The tympanum is widely open on the outer face proximal portion of the fore tibia (Fig. 11E, G).However, it differs from the species mentioned above in the longer tegmina, reaching up to the second abdominal tergite (Fig. 10C-D), which is more similar to Z. nauta.The first two segments of maxillary palpi are small and subequal in length; the third and fourth are as long as the first and second together; the fifth is longer than any other segment, with the ventral margin slightly arched and the medio-distal portion obliquely truncated, dorsal margin straight and the apex rounded.The only whitish area of the maxillary palpi is the ventral surface of the mediodistal portion; the remaining is dark (Fig. 11D).The abdomen is mostly black but has whitish bands on the anterior and posterior margins of each tergite (Fig. 10A-B).The supra-anal plate is subtriangular, with a distal margin rounded (Fig. 10E).The subgenital plate is similar to that of Z. nouragi, with the posterior margin sinuous in lateral view and concave in ventral view (Fig. 10F-G).The ovipositor is straight and very short, shorter than the hind tibia, with a triangular apex (Fig. 10A-B, 10H-I), similar to Z. guianensis.The copulatory papilla is quite unique, with an ovoid shape, laterally sinuous, and a more membranous dorsal portion (Fig. 11A-B).abdomen either (Figs 12A-B, 13E-H).Other differences easily noted are the longer tegmina (Fig. 12A-D), the large tympana on the outer faces of the fore tibia (Fig. 13E, in detail), and the tiny tympana on the inner face (Fig. 13F, in detail).The fore and hind (mid legs lost) legs have no whitish areas (Fig. 13E-F).The maxillary palpi are similar to the species described above but differ by the slightly concave fifth segment's dorsal surface and the slightly longer truncated-medio-distal portion (Fig. 13D).

New records
This is the first time that Zebragyllus has been recorded in Brazil.Two new species are being added to the genus, and two known species are recorded: Z. guianensis (Figs 14A-H, 16A) and Z. nouragui 16A).These two species were only known from their type locality, in French Guiana, where they live in sympatry, and here we extend their distribution (Fig. 16A).Five males and five females of Z. guianensis were collected in the Floresta Nacional de Caxiuanã, municipality of Melgaço, Pará State.Two males, two adult females, and one juvenil specimens of Z. nouragui were collected in Juruti, Pará State.It is plausible that these species occur in Brazil as these two localities are in Northern Pará State, near the boundary between Brazil and French Guiana, but on the other side of the great Amazon River.Floresta Nacional de Caxiuanã is about 630 km south of the Réserve Naturelle Nationale des Nourages (type locality), and Juruti is about 770 km southwest (Fig. 16A).The other species of Zebragryllus are only recorded in Peru and Colombia.So we expect these known species and new ones to also occur in Brazil as their distribution is unexplored.
The males of Z. nouragui studied here have a different chromatic pattern of the hind femora, a variation of the populations described by Desutter-Grandcolas et al. (2014).The paired whitish oblique bands described for the species, like those seen in the female hind femora (Fig. 15C), are absent, and only the ventral whitish band is displayed (Fig. 15A).
Species of Zebragryllus may occur in very low populations since just a few specimens were collected, even in considerable collection efforts, like in Floresta Nacional de Caxiuanã.This site was monitored for two years, with monthly armed pitfall traps in a 2-acre area.The area was divided into several parcels of 1 m², each with a pitfall trap installed in the center.Even so, only ten specimens of Z. guianensis were captured.

Discussion
The genus Zebragryllus and a potential move towards a cave-dwelling lifestyle Although cave orthopterans were initially a neglected fauna, some caves can host a diverse and endemic fauna of Orthoptera Olivier, 1789 and other taxa (Howarth 1972) and the observation of morphological regressive adaptations (troglomorphism) in their species stands out (Vandel 1965;Deharveng & Bedos 2018;Taylor 2019).These adaptations involve the presence of isolated or combined characteristics, such as eye reduction, loss of pigmentation, significant decrease in body size, wing reduction, and loss of tympanum compared to other species inhabiting surface environments (Vandel 1965;Desutter-Grandcolas et al. 1998;Di Russo & Sbordoni 1998;Deharveng & Bedos 2018).On the other hand, an increase in certain characteristics can also be observed, such as the elongation of appendages (legs and antennae) (Vandel 1965;Deharveng & Bedos 2018), the elongation of wings (Campos et al. 2023), and even an increase in body size (Vandel 1965;Carvalho et al. 2023).Additionally, it is important to emphasize that troglomorphism is not always related to troglobitic lifestyles (whether regressive or progressive), and the general theory of troglomorphism can lead to misguided interpretations of certain characters if not properly tested (see Desutter-Grandcolas 1997).The adaptations related to a cave-dwelling life need to be individually investigated (Desutter-Grandcolas 1997;Deharveng & Bedos 2018) once they may present different patterns depending on the biological entity being investigated, and natural selection, sexual selection, and/or genetic drift may potentially generate distinct patterns of morphological adaptations within the same group.
The presence of pre-adaptations in surface ancestors would be a first step towards colonizing cave environments, as demonstrated by Desutter-Grandcolas in a series of studies using Phalangopsidae as a model (see Desutter-Grandcolas et al. 1998).In this sense, an 'erroneous' analysis could consider as troglomorphic those characteristics that are actually pre-adaptations already present in surface ancestors.So, an investigation based on a phylogenetic context becomes a methodological requirement when the study focuses on crickets that have colonized caves (Desutter-Grandcolas 1997).
In the case of Zebragryllus aphonus Tavares, Oya & Cadena-Castañeda sp.nov., due to its discovery being based solely on collections in caves and upon observing for the first time the absence of a stridulatory apparatus, wing reduction, and different levels of tympanum development in this species, we may hypothesize, at least, two potential scenarios to be tested in the future: i) these characteristics represent adaptations selected in relation to the subterranean lifestyle of ferruginous cave-dwelling and a network of small channels (see Carmo & Kamino 2015;Ferreira et al. 2018); or ii) they are pre-adaptations selected related to a lifestyle in compact environments, such as leaf litter, tree cracks, and rocks.However, we still lack sufficient evidence, and the caves where this species was found are not extensively developed, often less than 10 meters in length.Thus, only evolutionary studies and descriptions of yet-unknown species within this genus will provide a better understanding of these findings and validate some of these hypotheses.

Minimum Convex Polygons for the genus Zebragryllus and distribution
Due to many locations in the Amazon Rainforest not having been inventoried or accurately studied from the taxonomic perspective of the genus Zebragryllus, there are still significant gaps in our taxonomic knowledge and geographic distribution of its species.To address this situation, we used the Minimum Convex Polygons map, which allowed us to identify these little-studied areas for the entire genus in a practical manner (Fig. 17).
Once all the known species records are placed in the Amazon rainforest or in transition zones between Amazon and Cerrado (Fig. 16), the polygon traced a probable distribution restricted to this biome, at most on the boundaries of the transition zones (Fig. 17).However, the known records undeniably reveal species from Oriental Amazonia and Occidental Amazonia.The two sets of points are too distant, and an enormous knowledge gap extends in the middle, a Wallacean shortfall (see Hortal et al. 2015).
Obviously, more collection efforts are needed to address this issue, and other new species will probably be discovered.

Fig. 16 .
Fig. 16.Distribution of species of Zebragryllus Desutter-Grandcolas & Cadena-Castañeda, 2014. A. In the Amazon rainforest of South America, with emphasis on the states of Pará and Tocantins, and the two municipalities that serve as new type localities.B-C.Limits of the Canaã dos Carajás municipality with a highlight on the region where seven caves are located, inhabited by Z. aphonus Tavares, Oya & Cadena-Castañeda sp.nov.D-E.Limits of the Conceição do Araguaia municipality with a highlight on the sampling site where Z. mebengokre Tavares, Oya & Cadena-Castañeda sp.nov.was found.