Open access journal: http://periodicos.uefs.br/index.php/sociobiology ISSN: 0361-6525 DOI: 10.13102/sociobiology.v69i4.8317Sociobiology 69(4): e8317 (December, 2022) The genus Metapolybia includes 18 species of social wasps of the tribe Epiponini with distribution restricted to the Neotropics (Andena et al., 2019), 16 of which are recorded in Brazil (Cooper, 1999; Andena & Carpenter, 2011; Andena et al., 2019; Somavilla et al., 2021). Wasps of this genus build colonies by swarming and build nests on different substrates (Souza & Zanuncio, 2012), without a pedicel and classified as astelocyttarus (Richards & Richards, 1951) and, for the most part, with a protective envelope (Cooper, 1999). Similarities between the chemical composition of social wasp nests and the plants on which they were built can provide information on the ideal physical conditions and safety and integrating the “chemical signature” of their colonies (Sguarizi-Antonio et al., 2021). Colony defense strategies by social wasps are diverse, including repellent substances against ants (Jeanne, 1970), Abstract Colony defense by social wasps includes aggressive behavior and camouflage with a high diversity of adaptation of their nests to the environment, including substrate choice. The objective of this study was to present new data on nesting and colony defense by the Neotropical social wasps Metapolybia cingulata and Metapolybia docilis (Hymenoptera: Vespidae). Field observations were made on January 4th and 5th, 2022 at the State Park of Iguaçu, state of Paraná, Brazil, on M. cingulata and M. docilis nests and information on nesting of these wasps obtained in 2014 at the Pandeiros River Wildlife Refuge (REVIS), and in 2012 at the Rio Doce State Park, state of Minas Gerais, Brazil. The types of substrates used by M. cingulata and M. docilis reinforce the importance of nest camouflage as a defense strategy for their colonies. The selection of nesting sites by these species was very variable. Sociobiology An international journal on social insects Marcos M. de Souza1, Norma Barbado2, José Adolfo M. de Almeida2, Gabriel Teofilo-Guedes3, José C. Zanuncio4 Article History Edited by Alexandre Somavilla, INPA, Brazil Received 30 June 2022 Initial acceptance 17 July 2022 Final acceptance 08 September 2022 Publication date 28 December 2022 Keywords Aggressive Displays, Colony Defense, Nest Architecture, Wasp Colony. Corresponding author José Adolfo M. de Almeida Instituto Federal de Educação, Ciência e Tecnologia do Paraná – IFPR Campus Umuarama, Parque Industrial, km 310, PR 323, CEP: 87507-014 Umuarama, Paraná, Brasil. E-Mail: jose.almeida@ifpr.edu.br nest built with protective envelope or camouflage and aggressive behavior (Chavarría-Pizzarro & West-Eberhard, 2010; Barbosa et al., 2016; Milani et al., 2021). Defense strategies of Metapolybia species include camouflage by homochromy and colony homotypy on tree trunks (Somavilla et al., 2012; Souza et al., 2020) and aggressive behavior in eventual disturbances, as reported for Metapolybia cingulata (Fabricius, 1804; Souza et al., 2020). The objective of this study was to present new data on nesting and colony defense in Neotropical social wasps Metapolybia cingulata (Fabricius, 1804) and Metapolybia docilis (Richards, 1978) (Hymenoptera: Vespidae). M. cingulata nests were recorded in 2010, in an evergreen forest in the Rio Doce State Park (19°42’ S, 42°34’ W) and the others in deciduous forest in the Pandeiros River Wildlife Refuge (REVIS) (15°88’ S, 45°95’ W), state of Minas 1 - Instituto Federal do Sul de Minas Gerais, Inconfidentes-MG, Brazil 2 - Instituto Federal do Paraná, Umuarama-PR, Brazil 3 - Universidade Estadual de Campinas, Campinas-SP, Brazil 4 - Universidade Federal de Viçosa, Viçosa-MG, Brazil Nest camouflage in Metapolybia cingulata and nesting and colony defensive behavior in Metapolybia docilis (Vespidae: Polistinae) in the Brazilian Atlantic Forest SHoRT NoTE mailto:jose.almeida@ifpr.edu.br Marcos M. de Souza, et al. – Nesting and defensive behavior of colonies of the wasps 2 Gerais in 2014. Nests of M. docilis were recorded in September 2014 and 2021 and in January 2022 in the State Park of Iguaçu (25°22’S, 54°2’W), state of Paraná, all in Brazil. Behavioral aspects of one colony of Metapolybia docilis were observed and recorded in the State Park of Iguaçu using the ad libitum method (Del-Claro, 2010) for 50 minutes divided into 10-minute sessions with two at 9:30 a.m., two at 5:00 p.m. and one at 1:00 p.m. on January 08th, 2022. This colony was subjected to stress stimuli: 1) manipulating the nest; 2) seeking to catch the specimens; 3) removing part of the nest envelope to model the behavior related to the aggressiveness or docility of Metapolybia. A part of the colony envelope (≈ 3 cm) was removed by exposing the comb (Chavarría-Pizzarro & West-Eberhard, 2010). The height of the other colonies made these procedures impossible. This study complied with the norms of the SISBIO (licence number: 76084-3). Each nest of M. cingulata and M. docilis was located in different plant species and anthropic substrates (Table 1). Data Species Metapolybia cingulata Metapolybia docilis Number of colonies Nine Four Locality State Park of Rio Doce (one nest); REVIS Rio Pandeiros (eight nests) State Park of Iguaçu Phytophysiognomy Evergreen forest (one nest); Deciduous Forest (eight nests). Evergreen forest Nesting Substrate Euterpe edulis (Arecaceae) (one nest); house construction lumber (eight nests). Trunk of Pliniarivularis (Cambess.) Myrtaceae (one nest); unidentified tree species (one nest), residential walls (two nests). Height above the ground 1.5 m (one nest); 2 to 2.4 m (eight nests). 2 m (one nest); 3.5 m (one nest); 4 and 5 m (two nests). Nest Color Light grey to dark gray, similar to substrate. Light grey to dark grey, similar to substrate. Table 1. Data on numbers of colonies, locality, phytophysiognomy nesting substrate, height above the ground and nest color of Metapolybia cingulata and Metapolybia docilis (Hymenoptera: Vespidae) colonies. Diversity of plant species used as substrate for nesting by M. docilis and M. cingulata were similar to those reported for Polistes actaeon Haliday, 1836 using from tree species, as Aspidosperma cuspa (Khunt), to shrubs, such as Baccharis dracunculifolia DC., and for Polybia platycephala Richards, 1978 and Protopolybia sedula (Saussure, 1854), nesting in nine and 19 plant species, respectively (Alvarenga et al., 2010; Souza et al., 2014). The M. cingulata and M. docilis nesting, on anthropic substrates, evidences the plasticity of nest site selection in Metapolybia species as reported for Mischocyttarus, Polistes and Polybia (Silva et al., 2019; Barbosa et al., 2020) in timber and walls (Oliveira et al., 2017; Silva et al., 2019) and Metapolybia miltoni Andena & Carpenter, 2011 (Silva et al., 2019), Metapolybia fraudator Carpenter and Andena, 2019 (Andena et al., 2019) and Metapolybia unilineata (Ihering, 1904) (Somavilla et al., 2012) nesting in wooden houses. Nesting in human constructions can offer greater protection against environmental factors, such as rainfall, wind and temperature extremes (Silva et al., 2022), which explains the use of these substrates near forested areas. The color of Metapolybia nests, similar to the substrate, seems to be related to homochromy camouflage, since in all the records made, there is a similarity between the color of the nest and the substrate (Fig 1), whether in a natural environment, using plant substrates, such as tree trunks or palm tree stems, and in anthropic environments, nesting on walls and wooden structures of houses, as already reported and suggested for species of the genera Mischocyttarus and Parachartergus (Souza et al., 2020), Metapolybia cingulata (Souza et al., 2020) and Chartergellus communis Richards, 1978 (Silva et al., 2022), which suggests that it constitutes a defense mechanism against visual predators, such as birds. The height of M. cingulata and M. docilis colonies on plant substrates was lower than those on anthropic substrates, the latter on roofs, and wooden or concrete structures with greater protection from the weather (Barbosa et al., 2020). This condition may be associated with the fact that nesting on anthropic substrates, below 2 m, could facilitate the destruction of nests by humans (López et al. 2012), however, this assumption needs to be better evaluated. Individuals of M. docilis, after stimulation by stress, behaved as follows: upon the first stimulus (touching with the hand or the handle of the entomological net), the individuals raised their heads and the first pair of legs, moving them intensely, but without attacking (Fig 2A), characterizing bluff behavior (Chavarría-Pizzarro & West-Eberhard, 2010). This may also characterize an alarm behavior, but no contraction of the gaster or wing movement was observed, as reported for Polistes erythrocephalus Latreille (1813) (West-Eberhard, 1969), Sociobiology 69(4): e8317 (December, 2022) 3 Polybia occidentalis (Olivier, 1791) (Jeanne, 1981) and Protopolybia exigua (Saussure, 1854) (Chadab, 1979). Alarm behavior was recorded in the morning, but not in the late afternoon, when the sun was directly shining on the colony, increasing wasp activity with foraging and colony maintenance (Giannotti et al., 1995; Santos & Presley, 2010). Wasp individuals subjected to the second (collection of individuals) and third (attempt to remove the colony envelope) stimuli raised their heads and the first pair of legs, moving them intensely, but they flew away or remained in the surroundings without attacking (Fig 2B). The absence of attack with only bluff behavior characterizes M. docilis as docile, increasing the importance of nest camouflage for this species (Chavarría-Pizzarro & West-Eberhard, 2010). However, the absence of sting attempts by this wasp differs from that reported for M. cingulata (Souza et al., 2020), showing that the colony defense behavior by Metapolybia species involves camouflage and, also, some degree of aggressiveness (Forsyth, 1981), as reported for Synoeca wasps (Chavarría-Pizarro & West-Eberhard, 2010). Camouflage as a defense strategy is important to select nesting substrates by Metapolybia cingulata and Metapolybia docilis and shows the plasticity in the selection of anthropic substrates or plants for nesting by these wasps. Fig 1. Nest camouflage in Metapolybia docilis (Hymenoptera: Vespidae) (a and b) at the State Park of Rio Doce; Metapolybia cingulata in the Pandeiros River Wildlife Refuge (REVIS) (c and d) and M. docilis in the State Park of Iguaçu (d, e and f), Brazil. Marcos M. de Souza, et al. – Nesting and defensive behavior of colonies of the wasps 4 Acknowledgements The authors thank IEF and SISBIO for providing license 76084-3. The taxonomist PhD Orlando Tobias Silveira, Museu Paraense Emílio Goeldi, who identified the collected specimens. The Itaipu Technological Park Foundation (FPTI) for sponsoring this study, to the Research sector of the Iguaçu National Park for hosting and to the Chico Mendes Biodiversity Institute (ICMBio), especially its brigade members for their logistical support. To the Brazilian institutions “Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)”, “Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES- Finance Code 001)”, “Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)”, and “Programa Cooperativo sobre Proteção Florestal (PROTEF) from Instituto de Pesquisas e Estudos Florestais (IPEF)” for financial support. Authors’ Contributions MMS: contributed on conceiving and designing the analysis, collecting the data, and writing both the draft and the final paper. NB: contributed on collecting the data, writing the draft and the final paper. JAMA: contributed on collecting the data, formatting, writing the draft and the final paper. GSTG: contributed on writing the draft and the final version, formatting, translating and preparing the figures. JCZ: contributed on designing the analysis, writing the draft and the final version, translating and supervising the manuscript writing. 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