Four new species of the genus Ochyrocera (Araneae, Ochyroceratidae) from iron caves of the state of Minas Gerais, with the description of the third anophtalmic species from Brazil

Four new species of the spider genus Ochyrocera Fage, 1912 are described from iron caves in the state of Minas Gerais, Brazil. Here we present the third anophtalmic and depigmented species of the genus, O. dorinha sp. nov., and three other depigmented species: O. monica sp. nov., O. magali sp. nov. and O. rosinha sp. nov. Only O. dorinha sp. nov. and O. rosinha sp. nov. are considered as troglobites due the ocular reduction or anophthalmia and elongated appendages, two troglomorphic features absent in the other species described.


Introduction
The known diversity of the spider family Ochyroceratidae in Brazil has recently increased by a relevant number of species from caves (Baptista et al. 2008;Brescovit et al. 2021). There are 16 species of Ochyrocera in Brazil (World Spider Catalog 2021), nine of which are from ferruginous caves in the state of Pará Brescovit et al. 2021), two from caves in the state of Minas Gerais (Baptista et al. 2008;, and the remaining five are epigean. Among the diversity in caves of the state of Minas Gerais, Ochyrocera ibitipoca Baptista, González & Tourinho, 2008 was described from a pseudo-karstic area composed of quartzite rocks in the region of Parque Estadual do Ibitipoca, in the south of the state (Baptista et al. 2008). Ochyrocera brumadinho , was found only in ferruginous caves in the municipality of Brumadinho in the region of the Quadrilátero Ferrífero (Iron Quadrangle) (Brescovit et al. 2021). Both species do not present any troglomorphism and share an accentuated olive-green color and six very characteristic eyes.
When reviewing specimens of Ochyroceratidae recently collected in iron caves of the state of Minas Gerais, we detected four new species with strong depigmentation of the body. We also found one of them to be an anophthalmic species, expanding the number of anophtalmic species of Ochyrocera to three, together with Ochyrocera ritxoco Brescovit, Zampaulo &Cizauskas, 2021 andO. ritxoo Brescovit, Zampaulo &Cizauskas, 2021, which are considered troglomorphic (Brescovit et al. 2021). In this work, we describe four new species of the genus Ochyrocera, all collected in ferruginous caves located in the Southern Serra do Espinhaço (Espinhaço Mountain Range) and/or in the Quadrilátero Ferrífero in the state of Minas Gerais in Southeast Brazil.

Study area
The four species described in this work are associated with caves inserted in the iron formations of two important regions of the state of Minas Gerais, the Quadrilátero Ferrífero and Serra do Espinhaço. Both regions comprise a large mosaic of phytophysiognomies shaped by the conjunction of topography, lithology, climate, and altitude (Jacobi and Carmo 2008) and are inserted in a transition zone between two Brazilian biodiversity hotspots, the Atlantic Forest and the Cerrado biomes (Mittermeier et al. 2004). In general, the climate in these regions is characterized as Cwb (High altitude subtropical climate), with mild and humid summers, and cool and dry winters (Köppen 1948), but it can be strongly influenced by the relief since the average altitude exceeds a thousand meters with the highest regions reaching two thousand meters. The annual precipitation varies between 1,250 and 1,550 mm and the average annual temperature between 18 °C and 19 °C.
Serra do Espinhaço comprises a chain of mountains about a thousand kilometers in length that extends from the north of the state of Bahia to the south center of the state of Minas Gerais, close to the Quadrilátero Ferrífero (Fig. 14). An important mining region since the colonial period, mainly due to the extraction of diamonds or gold, Serra do Espinhaço has been considered a World Biosphere Reserve since 2005, as it is one of the richest regions on the planet in terms of natural resources (UNESCO 2011). This mountain range also represents a major hydrographic divide in Southeast Brazil between the basins of the São Francisco River to the west, and the rivers that flow into the Atlantic Ocean to the east (Derby 1906;Saadi 1995). The species herein described from this region is found in the southern portion of the Southern Serra do Espinhaço (Fig. 14), where more than a thousand caves have been registered (CECAV 2021).
In turn, the Quadrilátero Ferrífero has an area of approximately 7,200km 2 and is considered one of the most important mineral provinces in Brazil, mainly due to its gold and iron deposits. At the same time, the region is one of the most floristically diverse areas in South America with high rates of endemism (Harley 1995;Giulietti et al. 1997). The area is considered to be of special biological relevance due to the presence of ferruginous fields, the occurrence of endemic plant species to the region, and because it constitutes a unique environment in the state. Formed by ancient and geologically complex terrains of the Minas Super Group, with varied lithologies (Alkmim and Marshak 1998;Klein and Ladeira 2000), more than two thousand caves are currently known in the region (CECAV 2021) and dozens of cave species have discovered and described in recent years.
Morphological terms follow , except for macrosetae of endites, which follows Baert (2014). Descriptions and measurements were performed using a LEICA 165C stereomicroscope. Photographs were taken with a Leica DFC 500 digital camera on a Leica MZ16A stereomicroscope. Focal-range images were made using Leica Application Suite software, version 2.5.0. Total and femur lengths were measured in lateral view without detaching any part from the specimen. All measurements are in millimeters. Female genitalia were excised with a sharp needle, digested using one tablet of enzymatic eye lens cleaner (Ultrazyme enzymatic cleaner) into 5ml distilled water for 24 hours and photographs were taken using Hoyer´s microscope slides. For scanning electron microscopy (SEM) images, body parts were dehydrated in a graded series of ethanol washes (80% to 100%), dried by critical point, mounted on metal stubs using adhesive copper tape and nail polish for fixation and covered with gold. SEM images were taken with a FEI Quanta 250 scanning electron microscope at Laboratório de Biologia Celular of Instituto Butantan, São Paulo, Brazil. Etymology. Noun in apposition is a tribute to the fictional character of the Brazilian "Turma da Mônica" comic books by Maurício de Sousa. Dorinha, created in 2004, is a visually impaired character, in this case blind. She was inspired by Dorina Nowill,  resembles O. magali sp. nov. by its long ducts and globose spermathecae (Fig. 7E, F) but differs by having a larger pore plate, and one single U-curve on the ducts of the spermathecae in the genitalia (Fig. 5C, D).
Female (paratype, IBSP 196428b). Total length 1.2; carapace length 0.5. Carapace as in male, yellowish-white (Figs 1B, 4A). Pedipalp without claw, with conical tip and subdistal tarsal organ (Fig. 4C, D). Eyes, chelicerae, sternum, endites, and labium as in male, but slightly yellowed (Fig. 4A, B). Legs yellowed; formula 4123, total lengths: I 4.6; II 4.4; III 4.3; IV 4.8. Abdomen length 0.6. Colulus triangular with five bristles. Triangular basal lateral pockets (Fig. 5C, D). Internal genitalia with globose spermathecae, with long and coiled ducts under pore plate. Uterus externus with no   (Fig. 14). In general, the species is found in larger caves for this type of lithology (above 30 meters of development), and with the presence of aphotic zones. Its type locality, cave MP_0008, is an important cave in the region, with numerous other troglobite species (Hoch and Ferreira 2012;Gomes et al. 2019), some of which are extremely rare (e.g., Ferricixius davidi Hoch & Ferreira, 2012;Cixiidae, Hemiptera). Inserted in a rupestrian field at an altitude of 1,500 meters, this cave has almost 100 meters of development, with ascending channels, high humidity and low availability of trophic resources. Etymology. Noun in apposition is a tribute to the fictional character of the Brazilian "Turma de Mônica" comic books by Maurício de Sousa, based on his daughter Magali. She is 7 years old and her main characteristic is her voracious appetite. She eats everything, at high speed, and feels hungry all the time, but despite this, friends see her as skinny, without ever putting on weight.

Ochyrocera magali
Diagnosis. Ochyrocera magali sp. nov. differs from other species of Ochyrocera by the male having conspicuous elongated setae on the border of the sternum (Fig. 6B, C), a long and apically projected embolus with curved tibiae of the male palp (Figs 6D, E,  8A, B). The female of Ochyrocera magali sp. nov. resembles O. dorinha sp. nov. by its long ducts and globose spermathecae (Fig. 5C, D) but differs by its small pore plate and not coiled ducts of the spermathecae in the genitalia and lateral pockets small and elongated (Fig. 7E, F).
Variation. No variation was found between the specimens. Distribution. Known only from four ferruginous caves in the municipalities of Caeté and Santa Bárbara, in the Quadrilátero Ferrífero, state of Minas Gerais, Brazil (Fig. 14). Etymology. Noun in apposition is a tribute to the fictional character of the Brazilian "Turma da Mônica" comic books by Maurício de Sousa. She is one of the main characters, along with her friend Cebolinha. She is a girl of strong genius, who has no patience for the nicknames she receives from other children because of her physical appearance and usually responds to such actions with her extreme brute strength, far superior to that of a girl her age.
Diagnosis. Ochyrocera monica sp. nov. is distinguished from the other species of Ochyrocera from the Quadrilátero Ferrífero by having cymbium with a conspicuous apical cuspule, enlarged palpal tibiae and elongated and sinuous embolus with a coiled tip (Fig. 9B-E). The female of Ochyrocera monica sp. nov. differs from other species by its large and elongated spermathecae and narrow and smooth columnar uterus externus (Fig. 9H, I).
Distribution. Known only from six iron caves located in the municipalities of Barão de Cocais and Mariana, in the Quadrilátero Ferrífero, state of Minas Gerais, Brazil (Fig. 14). Etymology. Noun in apposition is a tribute to the fictional character of the Brazilian "Turma da Mônica" comic books by Maurício de Sousa. She is a country girl, who is always wearing a red dress and a pair of pigtails in her hair. She never walks barefoot and she often speaks wrongly like a hick from the interior of Brazil.

Ochyrocera rosinha
Diagnosis. Ochyrocera rosinha sp. nov. is distinguished from the other species of Ochyrocera from the Quadrilátero Ferrífero by having a cymbium with a long apex, aciculiform cuspule, long and narrow palpal tibiae and filiform embolus, longer than cymbium (Figs 11C, D, 13A, B). The female of Ochyrocera rosinha sp. nov. differs from other species by its short and reniform spermathecae and very large pore plates (Fig. 13C, D).
Distribution. Known from the Lapa do Grotão cave system and SPT_0316 cave located in Southern Serra do Espinhaço, municipality of Morro do Pilar, state of Mi-nas Gerais, Brazil (Fig. 14). The Lapa do Grotão cave system, unlike the vast majority of ferruginous caves, is part of a perennial drainage system. This system is formed by two large caves (MP-0001A with 290 meters and MP-0001B with 451m) separated by a large doline (large circular doline with a diameter of approximately 45 m and a depth of 30 m). Both caves develop in the contact zone between iron formation rocks (itabirite) and siliciclastic rocks (quartzites). The system is located in an area of riparian forest and is associated with the Lages stream, which is mainly responsible for contributing organic matter to the underground environment. The system receives part of the flow from the Lages stream, which drains towards a sink located in its central west portion where there is a large well (Coelho and Leão 2015). In addition to O. rosinha sp. nov., several other troglomorphic species were found in this cave (R. Zampaulo, pers. obs.). Of these, only the harvestman species Gonycranaus pluto Bragagnolo, Hara & Pinto-da-Rocha, 2015 (Gerdesiidae, Opiliones) has been described so far.

Discussion
The discovery of a new anophthalmic species of Ochyrocera for the state of Minas Gerais, an area ca 2250 km from the state of Pará where the other two anophthalmic species were recently described (Brescovit et al. 2021), shows that the diversity of spiders with this morphological condition is greater than previously thought. At first sight, the condition of the total absence of eyes and depigmentation of Ochyrocera dorinha sp. nov. suggests an affinity with O. ritxoco Brescovit, Zampaulo &Cizauskas, 2021 andO. ritxoo Brescovit, Zampaulo &Cizauskas, 2021, species with the same body characteristics. However, close examination of genital structures, since the bodies are similar, suggests that O. dorinha sp. nov. is closer to O. magali sp. nov. a species from the same region. It is still difficult to establish relationships for Ochyrocera monica sp. nov. and O. rosinha sp. nov. without a review of all Neotropical species. However, both fit into the arietina group, which includes those species whose males have a palp with an entire cymbium and without a retrolateral apophysis .
All species described in this work have morphological characteristics related to the subterranean habitat (Romero 2009), such as body depigmentation. However, we believe that only O. rosinha sp. nov. and O. dorinha sp. nov. have specializations (ocular Figure 15. Ochyrocera dorinha sp. nov. A female B male C Landscape where cave MP_0008 is located (type locality). The yellow star represents the exact location where the cave is inserted D Mining located around cave MP_0008 E Entrance to cave MP_0008 F Internal aspect of cave MP_0008 (iron cave). reduction or anophthalmia and elongated appendages) that indicate restriction to this type of environment (troglobites).
The distribution of Ochyrocera dorinha sp. nov. is similar to the Tisentnops mineiro Brescovit & Sánchez-Ruiz, 2016, another troglobite spider located in ferruginous caves of the Quadrilátero Ferrífero (Brescovit & Sánchez-Ruiz 2016), although it is less abundant. Both species are distributed in caves in the areas of mining interest (Figs. 14, 15D) in the state of Minas Gerais, and lack studies aimed at the conservation of their populations.
Ochyrocera rosinha sp. nov. is not in an area of risk or threatened with extinction, and its population is restricted mainly to the caves of the Lapa do Grotão system (Fig. 16). This cave system is in a preserved environment due to legal protection by federal decree in Brazil since it is a large cave with rare troglobite species (Brasil 2008).
Although caves in iron formations are not large and extensive, they have been shown to be remarkably important in terms of animal life adapted to the subterranean environment (Ferreira et al. 2014). Spiders are very diverse in ferruginous systems in Brazil, and representatives of the genus Ochyrocera are, in addition to being diverse, specialized for life in this environment.