The Capixaba ant species inventory is far from complete : A local scale assessment of the ants ( Formicidae ) reveals the importance of diversity studies and entomological collections

Ants are considered ecosystem engineers for their essential environmental services provided (Del Toro et al., 2012). They occupy different habitat strata of urban ecosystems, from urban parks to buildings, or natural ecosystems from the canopy to the soil (Dáttilo et al., 2012; Santos-Silva et al., 2016; Vicente et al., 2016), interacting with several organisms such as animals, fungi, microbes, and plants, shaping the biodiversity (Sanders et al., 2014; Vicente et al., 2014; Pereira et al., 2015; Puker et al., 2015; Candiani & Bonaldo, 2017; Anjo-Pereira et al., 2021; Alencar et al., 2022; Santos et al., 2022). For these reasons, they react to environmental changes, therefore being excellent bioindicator Abstract The Atlantic Forest is the third largest biome in Brazil, the most sampled, and has the second highest diversity of ant species described. However, these data are spatially very heterogeneous, with the central region of the Atlantic Forest being better sampled. There is a visible gap from the south of Bahia to the north of Rio de Janeiro, comprising the entire state of Espírito Santo. Knowing this biodiversity gap, in this work, we list the ant fauna (Hymenoptera, Formicidae) deposited in the Coleção Entomológica of the Reserva Natural Vale (CERNV), located in the municipality of Linhares, Espírito Santo. The CERNV has 143 ants collected from August 1984, belonging to 63 species, 29 genera, and eight subfamilies. Of the 143 records, 94 are ant workers (66%), 27 queens (19%), and 22 males (15%). Seventy three percent of the ant workers are identified, 52% of queens and 36% of males. The species with the most individuals deposited are Camponotus sp. (15 records), Eciton burchellii (8), Atta sexdens (7), Neoponera villosa (7), and Solenopsis sp. (6). Among the identified ants, 12 species were recorded for the first time in the state of Espírito Santo, Brazil. The collection still holds workers (and probably unidentified males) of a species considered endangered, Dinoponera lucida. We can observe the importance of biological collections for the knowledge of biodiversity, local and regional since species are recorded for the first time in a state whose biome is widely studied, helping to fill in Linnean and Wallacean shortfalls. Sociobiology An international journal on social insects


Introduction
The Atlantic Forest is the third largest biome in Brazil, the most sampled, and which has the second highest diversity of ant species described Schmidt et al., 2022;Silva et al., 2022;Queiroz et al., 2023). The great and invaluable effort employed by researchers to create a dataset on Atlantic Forest ants generated a database with 53,818 ant records from 7,636 study locations . However, these data are spatially very heterogeneous, with the central region of the Atlantic Forest being better sampled (Schmidt et al., 2022). There is a visible gap from the south of Bahia to the north of Rio de Janeiro, comprising the entire state of Espírito Santo (Schmidt et al., 2022). This gap comprising the state of Espírito Santo is even more visible when we realize that it has 215 recorded species, and the neighboring state, Rio de Janeiro, with a similar size, has records of 485 species. Another neighboring state, Bahia, has more native species (603), reaching three times more registered species (645) in Minas Gerais (Janicki et al., 2016).
Knowing this biodiversity gap, in this work, we list the ant fauna (Hymenoptera, Formicidae) deposited in the Coleção Entomológica of the Reserva Natural Vale (CERNV), located in the municipality of Linhares, Espírito Santo. In addition, we highlight new records by state and discuss the importance of biological collections and increasing knowledge about the state's ant fauna.

Material and Methods
This study was conducted at the CERNV, located 30 km North of the Rio Doce. The collection receives specimens sampled at the Reserva Natural da Vale (RNV) by researchers who visit the reserve. The RNV covers part of the municipalities of Linhares and Jaguaré (19°06' -19°18'S and 39°45' -40°19'W - Figure 1), located in the North of the state of Espírito Santo, Brazil (Lima et al., 2022). The RNV encompasses approximately 23,000 hectares and is situated in one of the most important areas for the conservation of the biodiversity of the Atlantic Forest, being part of the Central Corridor of the Atlantic Forest and possessing one of the last great remnants of the Tabuleiro Forest, one of the most threatened formations of the Atlantic Forest biome (Reserva Natura Vale -Vale 2023). Since the mining company Vale began the process of acquiring the properties that gave rise to the RNV in the 1950s, researchers cataloged approximately 2,500 plant species, over 1,500 insect species or morphospecies, and 103 mammal species, in addition to 59 amphibians, 66 reptiles and 402 birds (Reserva Natura Vale -Vale 2023). The RNV has four distinct formations: the high forest, the Mussununga forest, the formations of floodable areas (herbaceous and forest), and the native fields (Vicens et al., 2004). The heterogeneity of the vegetation allows the formation of several ecosystems conducive to developing a great wealth of insect species and their preservation (Martins et al., 2014;Lima et al., 2022). Furthermore, RNV is surrounded by other conservation areas such as Reserva Biológica de Sooretama, RPPN Recanto das Antas, and RPPN Mutum Preto, which jointly reduce the effects of landscape modification in the region and have synergistic buffer effects (Martins-dos-Santos, 2023).
The ants were identified by four specialists, as listed in Table 1. We provide all the information on the label and the code associated with each specimen (Table 1). All specimens remain deposited in the CERNV.

Results
In the CERNV, 143 ant specimens have been deposited, collected between August 1984 and December 2013. The oldest samples date from August 8, 1984, and comprise 21 ants of different genera, sampled by Casimiro AB (Antonio B. Casimiro), Santos JS (José Simplício dos Santos), and Lacau S (Sébastien Lacau - Table 1). While the most recent record was collected on December 3rd, 2013.

Discussion
Considering the results of this study, we can observe the importance of biological collections for the knowledge of biodiversity, locally and regionally, since species are recorded for the first time in a state in which the Atlantic Forest biome is widely studied. Therefore, identifying the ants deposited in the Coleção Entomológica of the Reserva Natural da Vale helps fill in Linnean shortfalls by knowing the local fauna and Wallacean shortfalls by expanding the distribution of several species. The collection still holds workers (and probably unidentified males) of a species considered endangered, Dinoponera lucida (Fraga et al., 2019;Simon et al., 2020).
One exemplar of the ant D. lucida was sampled on December 10, 1986 (second oldest collection) by the retired professor at the Universidade Federal de Viçosa, Dr. Paulo Sergio Fiuza Ferreira. This Brazilian researcher has contributed strongly to the Coleção Entomológica of the RNV and to Brazilian entomology, mainly in the field of insect taxonomy with emphasis on Miridae (Hemiptera, Heteroptera. e.g., Carvalho & Ferreira, 1995;Ferreira et al., 2005;Ferreira & Henry, 2011). Dinoponera lucida is an endangered ant species endemic to the Atlantic Forest (Peixoto et al., 2010;Fraga et al., 2019;Silva et al., 2022). Recently, researchers studied the natural history the D. lucida in the RNV, observing its diurnal foraging that is relative to the litter leaf temperature, as well as its efficient care and maintenance of the nests, in addition to agonistic interactions representing the nest defense Zocca et al., 2021).
Other species that we highlight are the 12 species that are recorded for the first time in the state of Espírito Santo. Four species are from the genus Camponotus, which is one of the most diverse ant genera and has a challenging taxonomy (Baccaro et al., 2015). The Camponotus species that are new records, C. blandus, C. cingulatus, C. fastigatus, and C. melanoticus, are species with wide distribution in the Neotropical region, including neighboring states, such as the other new records belonging to other genera (Janicki, 2016), showing that the gap is due to the lack of studies in the state. Many of these species are from the oldest collections made by Antônio Batista Casimiro, Sébastien Lacau, and José Simplício dos Santos who was the curator at the time. Antônio Batista Casimiro, Sébastien Lacau, and other researchers from the municipality of Ilhéus, Bahia state, visited the RNV twice. The researchers used pitfall traps, random searches for ant nests, and nocturnal collections where they knocked plant branches into plastic bags.
The ants with the most records largely belong to genera with abundant colonies and large, easily collected species, such as Camponotus, Eciton, Atta, and Solenopsis (Baccaro et al., 2015). Except for Neoponera, which is a genus whose species have small colonies, including the sampled species N. villosa. Despite having colonies with less than 200 workers, N. villosa has been extensively studied and become a model system for studies of social behavior and foraging (Schmidt & Shattuck, 2014).
The proportion of castes in the collection reflects natural patterns, where reproductive forms are less abundant and seasonal. However, it is worth noting that we collected an undescribed queen of Pheidole fimbriata, highlighting the potential for new discoveries in the RNV. Since workers are the most abundant and frequent forms, the identification keys are based on workers and are not recommended for identifying queens and males Baccaro et al., 2015;Oliveira et al., 2021). This explains the pattern observed in the identification of ants found in the collection; after all, 73% of the workers were identified, in contrast to 52% of the queens and 36% of the males. Queens are generally more robust forms of workers, whereas males are quite different (Baccaro et al., 2015). Therefore, if the keys are about workers, association with workers sometimes can identify some queens, but never the males, which reinforces the fact that myrmecologists should invest in the manual collection of ants to access nests and associate castes .
Although the material deposited in the CERNV comes from sporadic, aleatory collections carried out by employees, and visitors at the RNV, without a systematized inventory using specific methodologies for the group, 19% of the sampled species are new records for the state of Espírito Santo. Therefore, carrying out a systematized inventory of the RNV, with methodologies that cover different microhabitats with the widest spatial coverage, would collaborate with the biodiversity of ants in the Espírito Santo state. Beyond that, it is crucial to know the history and heritage of the collections because, in addition to essential materials for the knowledge of biodiversity, they also carry a lot of local history, researchers, and Brazilian science (Ferreira et al., 2016;Andrade & Feitosa, 2020;Delabie et al., 2020;Fernandes & Oliveira, 2020;Oliveira et al., 2020;Prado et al., 2020;Souza-Campana et al., 2020;Ulysséa & Brandão, 2020).

Acknowledgments
We greatly appreciate Debora Mello Furtado de Mendonça, Sayonara Induzzi Cometti and Geovane Siqueira for their invaluable assistance during the visit to the Reserva Natural Vale. We greatly appreciate their promptness and patience in providing us with the necessary information about the history of CERNV. We thank Jacques Hubert Charles Delabie for identifying several ants listed in Table 1 and Victor Nagatani (University of Mogi das Cruzes) for identifying one of the specimens of Solenopsis virulens. LPP was supported by São Paulo Research Foundation (FAPESP) grant #2022/01974-8. JLPS and REV were supported by a CNPq PCI/INMA (respectively 300886/2022-5 and 301378/ 2023-1) post-doctoral scholarship.

Conflicts of Interest
The authors declare that they have no conflict of interest related to the publication of this manuscript.

Ethics
The authors confirm that the manuscript has been submitted only to this journal and confirm that all the research meets the ethical guidelines, including adherence to the legal requirements of the study country.

Data availability
The data is not yet available on digital platforms; however, all the data are reported in the manuscript, and the first author is available for any questions, or even the entomological collection where ants are deposited can be consulted.