Acoustic similarity between disjunct populations of the Marsh Tapaculo Scytalopus iraiensis Bornschein, Reinert & Pichorim, 1998 (Aves: Rhinocryptidae)

Acoustic similarity between disjunct populations of the Marsh Tapaculo Scytalopus iraiensis Bornschein, Reinert & Pichorim, 1998 (Aves: Rhinocryptidae) | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report Acoustic similarity between disjunct populations of the Marsh Tapaculo Scytalopus iraiensis Bornschein, Reinert & Pichorim, 1998 (Aves: Rhinocryptidae) José Alan Sales Souza, Ricardo Henrique Pereira da Silva, Bruno Pinho de Lucena, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6298732/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract This study investigated the vocalizations of Scytalopus iraiensis with the aim of identifying variations in vocalization between disjunct populations of the species. We analyzed 30 sound recordings of the species obtained from the Xeno-canto and Wiki Aves databases. The recordings were converted to the WAV format and then played back using Audacity software to obtain the following acoustic parameters: number of notes, duration, maximum frequency, and minimum frequency. After obtaining the parameters, Principal Component Analysis (PCA) was performed on the data in a smaller set of attributes. The PCA results were used in a correlation analysis, in which one of the variables that showed a high correlation (≥ 0.6) was excluded. After this process, the normality of the variables was verified using the Shapiro-Wilk test (p < 0.001), and those that did not meet the normality assumption were logarithmized. These data were associated with the records of each region and the similarity between the populations of the South and Southeast was assessed through a Permutational Multivariate Analysis of Variance (PERMANOVA). Our results showed that, despite the geographic isolation, the populations of S. iraiensis did not present significant variation in their vocalizations (RF = 0.7279, p = 0.425), indicating that the time they were separated was not enough to cause changes in their songs. Atlantic Forest Bioacoustics Biogeography Tyranni Figures Figure 1 Figure 2 Figure 3 Full Text The fragmentation of biological populations has the potential to cause divergence between individuals of a given species, whether genotypic, phenotypic, or behavioral, which can result in speciation (Haffer and Prance 2002; Repenning 2017; Lima and Mariano 2019). Sound communication is one of the factors that can differ due to regional adaptations and cultural mutations (Podos and Warren 2007; Cardoso 2016; Lima and Mariano 2019). Birds of the Tyranni Suborder (Passeriformes) tend not to diversify their vocalizations, which are typically innate and genetically transmitted, eliminating the need for learning (Dias 2009; Oliveira 2014). However, in a disjunct population, there is the possibility of sound segregation mechanisms being activated (Lima and Mariano 2019). Rhinocryptidae is a family of Neotropical birds comprising 12 genera, of which Scytalopus stands out for being the most diverse because of speciation processes in mountainous areas, especially in the Andes and Brazil (Sick 1997; Bornschein et al. 1998; Maurício et al. 2008). Their diversification began in the Middle Miocene (approximately 10 mya), when the oldest divergences in the family occurred, in which the lineages of Scytalopus diverged from their sister group ( Eleoscytalopus ) (Cadena et al. 2020). The second major diversification of Rhinocryptidae occurred at the end of the Pliocene (1.8-2.4 mya), giving rise to most of the species known today (Mata et al. 2009). At the beginning of the Pliocene, the territory that corresponds to southeastern Brazil had an arid and semi-arid climate, with humidity levels increasing only in the second half of this period (Grazziotin et al. 2006; Mata et al. 2009). As a result of the climate, forests have suffered considerable declines, fragmenting Rhinocryptidae. Considering that these species live in humid and mountainous regions, the environments that have maintained their suitable characteristics are located in areas with orographic rainfall, which has guaranteed greater stability over the years (Mata et al. 2009). Climatic fluctuations generate refuges in habitat patches throughout these fluctuations (Haffer and Prance 2002), and the current fragmented distribution of Scytalopus may be the result of climatic variations. Scytalopus iraiensis (Bornschein, Reinert & Pichorim, 1998), popularly known as Marsh Tapaculo (Del Hoyo et al. 2016; Pacheco 2021), is a bird that inhabits dense, closed vegetation associated with alluvial forests or river floodplains, occurring in the southern (Paraná, Santa Catarina, and Rio Grande do Sul) and southeastern (Minas Gerais) states of Brazil (Mata 2005; Klemann Jr. and Vieira 2013) (Fig. 1). It is classified as Endangered (EN) because of the destruction of its natural habitat. The song of S. iraiensis consists of a series of “tchek” notes, preceded by longer, more intonated notes and, the call, a “pic-pic” (BirdLife International 2016). Therefore, the aim of this study was to analyze the existence of differences in the vocalization of S. iraiensis , with an emphasis on disjunct populations of the species. In this study, a total of 30 sound recordings of S. iraiensis were analyzed, obtained from the Xeno-canto (https://xeno-canto.org) and Wiki Aves (https://www.wikiaves.com.br) databases. The selection of Xeno-canto recordings was based on the classifications provided by the website itself, considering only song-type vocalizations, with quality A or B, for analysis. To download them, the “querxc” function of the “warbleR” package (Araya-salas and Smith-Vidaurre 2017) was used in the R software (https://www.r-project.org). The records from Wiki Aves, in turn, were selected and downloaded manually, following the decreasing order of “Most popular” sounds of the species (Jun / 2023) and prioritizing recordings made in the Southeast region of the country, since these were scarce in Xeno-canto. After downloading, all the files obtained were converted from the original format (MP3) to WAV format, and then a manual analysis was carried out to detect and cut the best quality corners in each record, using the Audacity software (https://www.audacityteam.org) for such procedures. The first stage of data analysis consisted of collecting the variables of the songs, including parameters such as number of notes, duration, maximum frequency and minimum frequency. Afterwards, to reduce the number of variables that characterize these elements, a Principal Component Analysis (PCA) was used for the two subpopulations. Specifically, this technique allows the condensation of multiple characteristics into a smaller set of attributes, preserving the essential information, while reducing repetitiveness and data fluctuations (Jollands et al. 2004). With the results of the PCA (Fig. 2), a correlation analysis was performed, in which at least one variable that showed a high correlation rate (≥ 0.6) was excluded. Then, these variables had their normality verified using the Shapiro-Wilk test (p < 0.001), and those that did not fit the assumptions of normality were logarithmized. Finally, these data are associated with the recordings from each region and the similarity between the South and Southeast populations is assessed using a Permutational Multivariate Analysis of Variance (PERMANOVA), an analysis used to test multivariate hypotheses comparing species to environmental variables (Anderson and Braak, 2003; Da Silva et al. 2022) The song-like vocalizations of S. iraiensis analyzed consist of a series of 49 to 1025 monosyllabic notes, with a frequency band ranging from 0.5 kHz to 4.4 kHz, an average dominant frequency of 2.2 kHz, and an average duration of 68.2 s, ranging from 12.8 to 223.4 s (Fig. 3). Among the four variables applied by the semi-automated function of the “warbleR” package, three were selected for the tests, which culminated in a PERMANOVA, whose results (RF = 0.7279; p = 0.425) revealed no significant differences between the subpopulations, suggesting that the separation time between them was not sufficient to cause relevant changes in the song. Although disjunction between populations can generate different selective pressures, leading to divergence in functional traits, such as changes in vocalizations and the formation of distinct groups (Räsänen and Hendry, 2008; Hermes et al., 2016), such an effect was not observed in the subpopulations of S. iraiensis . These subpopulations maintained uniformity in song structure, coloration pattern and morphometry (Vasconcelos et al., 2008a). One possible explanation for these observations is that, although there are no records of the species in the territory corresponding to the state of São Paulo to date, it has ecological zones with plant formations that are conducive to the occurrence of the species, such as the Serra do Mar State Park (Vasconcelos et al. 2008a, b; Klemann Jr. and Vieira 2013). Considering that these characteristics are present in the territory adjacent to the areas where the species occurs, gene flow may occur through migrations between subpopulations. However, further studies are still needed. This result is consistent with that obtained by Lima and Mariano (2019), who investigated disjunct populations of Formicarius colma Boddaert, 1783, also belonging to the Suborder Tyranni, whose diversification occurred approximately 6.5 million years ago. These are subpopulations distributed north of the Amazon River and along the Atlantic Forest and, despite the even older geographical separation between the Amazon Forest and the Atlantic Forest, no significant variations in their song were evidenced (Lima and Mariano 2019). Thus, although no variations were observed in the vocalizations of the studied subpopulations, our results, in line with those of Lima and Mariano (2019), reinforce the idea that the song of birds of the Suborder Tyranni has a predominantly innate nature. Declarations Acknowledgments We would like to thank Adjane Medeiros de Souza for her valuable contribution in creating and providing the map "Locations of Sound Records of Scytalopus iraiensis " (Fig. 1), which was fundamental to the quality and accuracy of this article. Author contributions Conceptualization: José Alan Sales Souza; Methodology: José Alan Sales Souza, Bruno Pinho de Lucena, Erich de Freitas Mariano; Formal analysis and investigation: José Alan Sales Souza, Ricardo Henrique Pereira da Silva, Bruno Pinho de Lucena, Gabriel Rian Alexandre Ferreira; Writing - original draft preparation: José Alan Sales Souza, Ricardo Henrique Pereira da Silva, Bruno Pinho de Lucena, Gabriel Rian Alexandre Ferreira; Writing - review and editing: Bruno Pinho de Lucena, Gabriel Rian Alexandre Ferreira; Supervision: Erich de Freitas Mariano. Ethics approval: None. Conflict of interest: The authors declare no competing interests. Funding : No funding, grants, or other support was received. Financial interests: The authors declare they have no financial interests. Consent: All authors consent to participate in and publish this study. References Anderson M, Braak CT (2003) Permutation tests for multi-factorial analysis of variance. Journal of statistical computation and simulation, 73(2):85-113. https://doi.org/10.1080/00949650215733 Araya-Salas M, Smith-Vidaurre G (2017) WarbleR: An R Package to Streamline Analysis of Animal Acoustic Signals. Methods in Ecology and Evolution, 8(2):184-191. https://doi.org/10.1111/2041-210X.12624 BirdLife International (2016) Scytalopus iraiensis . The IUCN Red List of Threatened Species 2016: e.T22728464A94986972. http://dx.doi.org/10.2305/IUCN.UK.2016-3.RLTS.T22728464A94986972.en Bornschein MR, Reinert BL, Pichorim M (1998) Descrição, Ecologia e Conservação de um Novo Scytalopus (Rhinocryptidae) do Sul do Brasil, com Comentários Sobre a Morfologia da Família. Ararajuba, 6(1):3-36. Cadena CD, Cuervo AM, Céspedes LN, Bravo GA, Krabbe N, Schulenberg TS, Derryberry GE, Silveira LF, Derryberry EP, Brumfield RT, Fjeldså J (2020) Systematics, Biogeography, and Diversification of Scytalopus Tapaculos (Rhinocryptidae), an Enigmatic Radiation of Neotropical Montane Birds. The Auk, 137(2):1-30. https://doi.org/10.1093/auk/ukaa024 Cardoso GS (2016) Variação Bioacústica das Vocalizações do Complexo Taraba major (Vieillot, 1816) (Aves: Passeriformes: Thamnophilidae). Dissertação, Universidade Estadual Paulista “Júlio de Mesquita Filho”. Da Silva FR, Gonçalves-Souza T, Paterno GB, Provete DB, Vancine MH (2022) Análises ecológicas no R. Nupeea, Recife. De Vasconcelos MF, Lopes LE, Machado CG, Rodrigues M (2008b) As aves dos campos rupestres da Cadeia do Espinhaço: diversidade, endemismo e conservação. Megadiversidade, 4(1-2):221-241. De Vasconcelos MF, Mauricio GN, Kirwan GM, Silveira LF (2008a) Range extension for Marsh Tapaculo Scytalopus iraiensis to the highlands of Minas Gerais, Brazil, with an overview of the species' distribution. Bull. B.O.C., 128(2):101-106. Del Hoyo J, Collar NJ, Christie DA, Elliott A, Fishpool LDC, Boesman P, Kirwan GM 2016. Illustrated Checklist of the Birds of the World. Volume 2: Passerines. Lynx Edicions, Barcelona, Spain and Cambridge. Dias AFS (2009) Comparação e Descrição de Parâmetros Acústicos do Canto da Volatinia jacarina (Aves: Emberezidae) no Contexto de Seleção Sexual. Dissertação, Universidade de Brasília. Grazziontin FG, Monzel M, Echeverrigaray S, Bonatto SL (2006) Phylogeography of the Bothrops jararaca complex (Serpentes: Viperidae): Past Fragmentation and Island Colonization in the Brazilian Atlantic Forest. Molecular Ecology, 15(13):3969-3982. https://doi.org/10.1111/j.1365-294X.2006.03057.x Haffer J, Prance GT (2002) Impulsos Climáticos da Evolução na Amazônia Durante o Cenozóico: Sobre a Teoria dos Refúgios da Diferenciação Biótica. Estudos Avançados, 16(46):175-206. https://doi.org/10.1590/S0103-40142002000300014 Hermes C, Döpper A, Schaefer HM, Segelbacher G (2016) Effects of forest fragmentation on the morphological and genetic structure of a dispersal-limited, endangered bird species. Nature Conservation, 16:39-58. https://doi.org/10.3897/natureconservation.16.10905 Jollands N, Lermit J, Patterson M (2004) Aggregate eco-efficiency indices for New Zealand—a principal components analysis. Journal of Environmental Management, 73(4):293-305. https://doi.org/10.1016/j.jenvman.2004.07.002 Klemann JRL, Vieira JS (2013) Assessing the extent of occurrence, area of occupancy, territory size, and population size of Marsh Tapaculo ( Scytalopus iraiensis ). Animal Biodiversity and Conservation, 36(1):47-57. Lima MC, Mariano EF (2019) Acoustic similarity among disjunct populations of Formicarius colma Boddaert, 1783 (Aves: Formicariidae). Ciência e Natura, Santa Maria, 41:1-7. https://doi.org/10.5902/2179460X33207 Mata H (2005) História evolutiva das espécies não-andinas de Scytalopus inferida através da variabilidade no DNA mitocondrial. Dissertação, Pontifícia Universidade Católica do Rio Grande do Sul. Mata H, Fontana CS, Maurício GN, Bornschein MR, Vasconcelos MF, Bonatto SL (2009) Molecular phylogeny and biogeography of the eastern Tapaculos (Aves: Rhinocryptidae: Scytalopus , Eleoscytalopus ): Cryptic diversification in Brazilian Atlantic Forest. Molecular Phylogenetics and Evolution, 53(2):450-462. https://doi.org/10.1016/j.ympev.2009.07.017 Maurício GN, Mata N, Bornschein MR, Cadena CD, Alvarenga H, Bonatto SL (2008) Hidden generic diversity in Neotropical birds: Molecular and anatomical data support a new genus for the ‘‘Scytalopus” indigoticus species-group (Aves: Rhinocryptidae). Molecular Phylogenetics and Evolution, 49(1):125-135. https://doi.org/10.1016/j.ympev.2008.06.017 Oliveira VLC (2014) Influência do ruído ambiente em canções de um pássaro Oscine e um Suboscine da mesma população. Dissertação, Universidade Federal de Alfenas. Pacheco JF, Silveira LF, Aleixo A, Agne CE (2021) Annotated checklist of the birds of Brazil by the Brazilian Ornithological Records Committee—second edition. Ornithology Research, 29:94-105. https://doi.org/10.1007/s43388-021-00058-x Podos J, Warren PS (2007) The evolution of geographic variation in birdsong. Advances in the study of behavior or Elsevier, 37:403-458. https://doi.org/10.1016/S0065-3454(07)37009-5 Räsänen K, Hendry AP (2008) Disentangling interactions between adaptive divergence and gene flow when ecology drives diversification. Ecology letters, 11(6):624-636. https://doi.org/10.1111/j.1461-0248.2008.01176.x Repenning M (2017) Variação geográfica em Sporophila (Aves: Thraupidae) com base em evidências fenotípicas, ecológicas e genéticas. Tese, Pontifícia Universidade Católica do Rio Grande do Sul. Sick H (1997) Ornitologia Brasileira. Nova Fronteira, Rio de Janeiro. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6298732","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":453978388,"identity":"a2ff1161-6c2b-447f-9cbe-9ec341f51470","order_by":0,"name":"José Alan Sales Souza","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABCElEQVRIiWNgGAWjYDACdjBpAWQ0tjEkMNgAOYyNB/BqYQaTEgwMPIdBWtJAWhqI1CKR3gZkHAZz8Wrhb2Z+9vBHjYScfENi24MHFeft1rYfBtpSYxONS4vEYTZzY55jEsYGBw62GyScuZ287UwiUMuxtNwGXHoOM5hJM7BJJG5gbGyTSGy7nWx2AKiFseEwTi3yh9m/Sf74J1E/v5kRpOVcstn5h/i1GBzmMZPgbZNIYDgG1nLAzuwGAVsMD/OUSfP2SRhuOMMI8ktygtkNoC0JePwid7x9m+SPbzby8vOfA4Ouws7e7Hz6wwcfamxwex8dJIJVJhCrHATsSVE8CkbBKBgFIwMAAB70Yr1NiWzAAAAAAElFTkSuQmCC","orcid":"","institution":"Universidade Federal de Campina Grande","correspondingAuthor":true,"prefix":"","firstName":"José","middleName":"Alan Sales","lastName":"Souza","suffix":""},{"id":453978389,"identity":"f14fe27a-2059-4a24-b893-bf70d2388467","order_by":1,"name":"Ricardo Henrique Pereira da Silva","email":"","orcid":"","institution":"Universidade Federal de Campina Grande","correspondingAuthor":false,"prefix":"","firstName":"Ricardo","middleName":"Henrique Pereira da","lastName":"Silva","suffix":""},{"id":453978390,"identity":"973916b1-a8d2-4255-af3f-69a7e58f2ae7","order_by":2,"name":"Bruno Pinho de Lucena","email":"","orcid":"","institution":"Universidade Federal de Campina Grande","correspondingAuthor":false,"prefix":"","firstName":"Bruno","middleName":"Pinho","lastName":"de Lucena","suffix":""},{"id":453978391,"identity":"21006137-ea60-4c8d-bc30-84f6347243d6","order_by":3,"name":"Gabriel Rian Alexandre Ferreira","email":"","orcid":"","institution":"Universidade Federal de Campina Grande","correspondingAuthor":false,"prefix":"","firstName":"Gabriel","middleName":"Rian Alexandre","lastName":"Ferreira","suffix":""},{"id":453978392,"identity":"27e41d09-3bf0-4635-bd82-41d8c7776c55","order_by":4,"name":"Erich de Freitas Mariano","email":"","orcid":"","institution":"Universidade Federal de Campina Grande","correspondingAuthor":false,"prefix":"","firstName":"Erich","middleName":"de Freitas","lastName":"Mariano","suffix":""}],"badges":[],"createdAt":"2025-03-25 00:08:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6298732/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6298732/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":82362551,"identity":"f6645210-fe6f-4093-a378-1758e7be4ff5","added_by":"auto","created_at":"2025-05-09 12:08:17","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":2172088,"visible":true,"origin":"","legend":"\u003cp\u003eLocations of Sounds Records of \u003cem\u003eScytalopus iraiensis\u003c/em\u003e. (\u003cstrong\u003eRed dots\u003c/strong\u003e) Population in the Southeast region. (\u003cstrong\u003eYellow dots\u003c/strong\u003e) Population in the South region.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6298732/v1/3dd22ac71395c5c05e574eb5.png"},{"id":82362556,"identity":"2ecdeb6d-7b5e-4fec-83c0-c1503fc28599","added_by":"auto","created_at":"2025-05-09 12:08:17","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":218619,"visible":true,"origin":"","legend":"\u003cp\u003ePCA result: correlation between the bioacoustic parameters analyzed in the songs of \u003cem\u003eScytalopus iraiensis\u003c/em\u003e. (\u003cstrong\u003eDur\u003c/strong\u003e) Vocalization duration. (\u003cstrong\u003eNnotas\u003c/strong\u003e) Number of notes. (\u003cstrong\u003eFFmax\u003c/strong\u003e) Maximum frequency. (\u003cstrong\u003eFFmin\u003c/strong\u003e) Minimum frequency.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6298732/v1/a0315f8d64f8f9417bfe6c9c.png"},{"id":82362548,"identity":"3ca47e75-9246-447f-9b02-bc669783cb43","added_by":"auto","created_at":"2025-05-09 12:08:17","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1172746,"visible":true,"origin":"","legend":"\u003cp\u003eSpectrogram of the song of \u003cem\u003eScytalopus iraiensis\u003c/em\u003e. The X-axis represents Time (s), and the Y-axis represents Frequency (kHz).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6298732/v1/591c161dd9030f5d32c78bac.png"},{"id":94186734,"identity":"7582ba53-2f43-47bf-82e0-a1994e086a36","added_by":"auto","created_at":"2025-10-23 11:02:01","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4501668,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6298732/v1/259960b7-5ddf-409d-bb3b-a5c909778a95.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Acoustic similarity between disjunct populations of the Marsh Tapaculo Scytalopus iraiensis Bornschein, Reinert \u0026 Pichorim, 1998 (Aves: Rhinocryptidae)","fulltext":[{"header":"Full Text","content":"\u003cp\u003eThe fragmentation of biological populations has the potential to cause divergence between individuals of a given species, whether genotypic, phenotypic, or behavioral, which can result in speciation (Haffer and Prance 2002; Repenning 2017; Lima and Mariano 2019). Sound communication is one of the factors that can differ due to regional adaptations and cultural mutations (Podos and Warren 2007; Cardoso 2016; Lima and Mariano 2019). Birds of the Tyranni Suborder (Passeriformes) tend not to diversify their vocalizations, which are typically innate and genetically transmitted, eliminating the need for learning (Dias 2009; Oliveira 2014). However, in a disjunct population, there is the possibility of sound segregation mechanisms being activated (Lima and Mariano 2019).\u003c/p\u003e\n\u003cp\u003eRhinocryptidae is a family of Neotropical birds comprising 12 genera, of which \u003cem\u003eScytalopus\u003c/em\u003e stands out for being the most diverse because of speciation processes in mountainous areas, especially in the Andes and Brazil (Sick 1997; Bornschein et al. 1998; Maur\u0026iacute;cio et al. 2008). Their diversification began in the Middle Miocene (approximately 10 mya), when the oldest divergences in the family occurred, in which the lineages of \u003cem\u003eScytalopus\u003c/em\u003e diverged from their sister group (\u003cem\u003eEleoscytalopus\u003c/em\u003e) (Cadena et al. 2020). The second major diversification of Rhinocryptidae occurred at the end of the Pliocene (1.8-2.4 mya), giving rise to most of the species known today (Mata et al. 2009).\u003c/p\u003e\n\u003cp\u003eAt the beginning of the Pliocene, the territory that corresponds to southeastern Brazil had an arid and semi-arid climate, with humidity levels increasing only in the second half of this period (Grazziotin et al. 2006; Mata et al. 2009). As a result of the climate, forests have suffered considerable declines, fragmenting Rhinocryptidae. Considering that these species live in humid and mountainous regions, the environments that have maintained their suitable characteristics are located in areas with orographic rainfall, which has guaranteed greater stability over the years (Mata et al. 2009). Climatic fluctuations generate refuges in habitat patches throughout these fluctuations (Haffer and Prance 2002), and the current fragmented distribution of Scytalopus may be the result of climatic variations.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eScytalopus iraiensis\u003c/em\u003e (Bornschein, Reinert \u0026amp; Pichorim, 1998), popularly known as Marsh Tapaculo (Del Hoyo et al. 2016; Pacheco 2021), is a bird that inhabits dense, closed vegetation associated with alluvial forests or river floodplains, occurring in the southern (Paran\u0026aacute;, Santa Catarina, and Rio Grande do Sul) and southeastern (Minas Gerais) states of Brazil (Mata 2005; Klemann Jr. and Vieira 2013) (Fig. 1). It is classified as Endangered (EN) because of the destruction of its natural habitat. The song of \u003cem\u003eS. iraiensis\u003c/em\u003e consists of a series of \u0026ldquo;tchek\u0026rdquo; notes, preceded by longer, more intonated notes and, the call, a \u0026ldquo;pic-pic\u0026rdquo; (BirdLife International 2016). Therefore, the aim of this study was to analyze the existence of differences in the vocalization of \u003cem\u003eS. iraiensis\u003c/em\u003e, with an emphasis on disjunct populations of the species.\u003c/p\u003e\n\u003cp\u003eIn this study, a total of 30 sound recordings of \u003cem\u003eS. iraiensis\u003c/em\u003e were analyzed, obtained from the Xeno-canto (https://xeno-canto.org) and Wiki Aves (https://www.wikiaves.com.br) databases. The selection of Xeno-canto recordings was based on the classifications provided by the website itself, considering only song-type vocalizations, with quality A or B, for analysis. To download them, the \u0026ldquo;querxc\u0026rdquo; function of the \u0026ldquo;warbleR\u0026rdquo; package (Araya-salas and Smith-Vidaurre 2017) was used in the R software (https://www.r-project.org). The records from Wiki Aves, in turn, were selected and downloaded manually, following the decreasing order of \u0026ldquo;Most popular\u0026rdquo; sounds of the species (Jun / 2023) and prioritizing recordings made in the Southeast region of the country, since these were scarce in Xeno-canto. After downloading, all the files obtained were converted from the original format (MP3) to WAV format, and then a manual analysis was carried out to detect and cut the best quality corners in each record, using the Audacity software (https://www.audacityteam.org) for such procedures.\u003c/p\u003e\n\u003cp\u003eThe first stage of data analysis consisted of collecting the variables of the songs, including parameters such as number of notes, duration, maximum frequency and minimum frequency. Afterwards, to reduce the number of variables that characterize these elements, a Principal Component Analysis (PCA) was used for the two subpopulations. Specifically, this technique allows the condensation of multiple characteristics into a smaller set of attributes, preserving the essential information, while reducing repetitiveness and data fluctuations (Jollands et al. 2004). With the results of the PCA (Fig. 2), a correlation analysis was performed, in which at least one variable that showed a high correlation rate (\u0026ge; 0.6) was excluded. Then, these variables had their normality verified using the Shapiro-Wilk test (p \u0026lt; 0.001), and those that did not fit the assumptions of normality were logarithmized.\u003c/p\u003e\n\u003cp\u003eFinally, these data are associated with the recordings from each region and the similarity between the South and Southeast populations is assessed using a Permutational Multivariate Analysis of Variance (PERMANOVA), an analysis used to test multivariate hypotheses comparing species to environmental variables (Anderson and Braak, 2003; Da Silva et al. 2022)\u003c/p\u003e\n\u003cp\u003eThe song-like vocalizations of \u003cem\u003eS. iraiensis\u003c/em\u003e analyzed consist of a series of 49 to 1025 monosyllabic notes, with a frequency band ranging from 0.5 kHz to 4.4 kHz, an average dominant frequency of 2.2 kHz, and an average duration of 68.2 s, ranging from 12.8 to 223.4 s (Fig. 3). Among the four variables applied by the semi-automated function of the \u0026ldquo;warbleR\u0026rdquo; package, three were selected for the tests, which culminated in a PERMANOVA, whose results (RF = 0.7279; p = 0.425) revealed no significant differences between the subpopulations, suggesting that the separation time between them was not sufficient to cause relevant changes in the song.\u003c/p\u003e\n\u003cp\u003eAlthough disjunction between populations can generate different selective pressures, leading to divergence in functional traits, such as changes in vocalizations and the formation of distinct groups (R\u0026auml;s\u0026auml;nen and Hendry, 2008; Hermes et al., 2016), such an effect was not observed in the subpopulations of \u003cem\u003eS. iraiensis\u003c/em\u003e. These subpopulations maintained uniformity in song structure, coloration pattern and morphometry (Vasconcelos et al., 2008a).\u003c/p\u003e\n\u003cp\u003eOne possible explanation for these observations is that, although there are no records of the species in the territory corresponding to the state of S\u0026atilde;o Paulo to date, it has ecological zones with plant formations that are conducive to the occurrence of the species, such as the Serra do Mar State Park (Vasconcelos et al. 2008a, b; Klemann Jr. and Vieira 2013). Considering that these characteristics are present in the territory adjacent to the areas where the species occurs, gene flow may occur through migrations between subpopulations. However, further studies are still needed.\u003c/p\u003e\n\u003cp\u003eThis result is consistent with that obtained by Lima and Mariano (2019), who investigated disjunct populations of \u003cem\u003eFormicarius colma\u003c/em\u003e Boddaert, 1783, also belonging to the Suborder Tyranni, whose diversification occurred approximately 6.5 million years ago. These are subpopulations distributed north of the Amazon River and along the Atlantic Forest and, despite the even older geographical separation between the Amazon Forest and the Atlantic Forest, no significant variations in their song were evidenced (Lima and Mariano 2019).\u003c/p\u003e\n\u003cp\u003eThus, although no variations were observed in the vocalizations of the studied subpopulations, our results, in line with those of Lima and Mariano (2019), reinforce the idea that the song of birds of the Suborder Tyranni has a predominantly innate nature.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe would like to thank Adjane Medeiros de Souza for her valuable contribution in creating and providing the map \u0026quot;Locations of Sound Records of \u003cem\u003eScytalopus iraiensis\u003c/em\u003e\u0026quot; (Fig. 1), which was fundamental to the quality and accuracy of this article.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConceptualization: Jos\u0026eacute; Alan Sales Souza; Methodology: Jos\u0026eacute; Alan Sales Souza, Bruno Pinho de Lucena, Erich de Freitas Mariano; Formal analysis and investigation: Jos\u0026eacute; Alan Sales Souza, Ricardo Henrique Pereira da Silva, Bruno Pinho de Lucena, Gabriel Rian Alexandre Ferreira; Writing - original draft preparation: Jos\u0026eacute; Alan Sales Souza, Ricardo Henrique Pereira da Silva, Bruno Pinho de Lucena, Gabriel Rian Alexandre Ferreira; Writing - review and editing: Bruno Pinho de Lucena, Gabriel Rian Alexandre Ferreira; Supervision: Erich de Freitas Mariano.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval:\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNone.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest:\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e:\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNo funding, grants, or other support was received.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFinancial interests:\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe authors declare they have no financial interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent:\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAll authors consent to participate in and publish this study.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAnderson M, Braak CT (2003) Permutation tests for multi-factorial analysis of variance. 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Disserta\u0026ccedil;\u0026atilde;o, Universidade Estadual Paulista \u0026ldquo;J\u0026uacute;lio de Mesquita Filho\u0026rdquo;.\u003c/li\u003e\n \u003cli\u003eDa Silva FR, Gon\u0026ccedil;alves-Souza T, Paterno GB, Provete DB, Vancine MH (2022) An\u0026aacute;lises ecol\u0026oacute;gicas no R. Nupeea, Recife.\u003c/li\u003e\n \u003cli\u003eDe Vasconcelos MF, Lopes LE, Machado CG, Rodrigues M (2008b) As aves dos campos rupestres da Cadeia do Espinha\u0026ccedil;o: diversidade, endemismo e conserva\u0026ccedil;\u0026atilde;o. Megadiversidade, 4(1-2):221-241.\u003c/li\u003e\n \u003cli\u003eDe Vasconcelos MF, Mauricio GN, Kirwan GM, Silveira LF (2008a) Range extension for Marsh Tapaculo \u003cem\u003eScytalopus iraiensis\u003c/em\u003e to the highlands of Minas Gerais, Brazil, with an overview of the species\u0026apos; distribution. Bull. B.O.C., 128(2):101-106.\u003c/li\u003e\n \u003cli\u003eDel Hoyo J, Collar NJ, Christie DA, Elliott A, Fishpool LDC, Boesman P, Kirwan GM 2016. Illustrated Checklist of the Birds of the World. Volume 2: Passerines. Lynx Edicions, Barcelona, Spain and Cambridge.\u003c/li\u003e\n \u003cli\u003eDias AFS (2009) Compara\u0026ccedil;\u0026atilde;o e Descri\u0026ccedil;\u0026atilde;o de Par\u0026acirc;metros Ac\u0026uacute;sticos do Canto da \u003cem\u003eVolatinia jacarina\u003c/em\u003e (Aves: Emberezidae) no Contexto de Sele\u0026ccedil;\u0026atilde;o Sexual. Disserta\u0026ccedil;\u0026atilde;o, Universidade de Bras\u0026iacute;lia.\u003c/li\u003e\n \u003cli\u003eGrazziontin FG, Monzel M, Echeverrigaray S, Bonatto SL (2006) Phylogeography of the \u003cem\u003eBothrops jararaca\u003c/em\u003e complex (Serpentes: Viperidae): Past Fragmentation and Island Colonization in the Brazilian Atlantic Forest. Molecular Ecology, 15(13):3969-3982. https://doi.org/10.1111/j.1365-294X.2006.03057.x\u003c/li\u003e\n \u003cli\u003eHaffer J, Prance GT (2002) Impulsos Clim\u0026aacute;ticos da Evolu\u0026ccedil;\u0026atilde;o na Amaz\u0026ocirc;nia Durante o Cenoz\u0026oacute;ico: Sobre a Teoria dos Ref\u0026uacute;gios da Diferencia\u0026ccedil;\u0026atilde;o Bi\u0026oacute;tica. Estudos Avan\u0026ccedil;ados, 16(46):175-206. https://doi.org/10.1590/S0103-40142002000300014\u003c/li\u003e\n \u003cli\u003eHermes C, D\u0026ouml;pper A, Schaefer HM, Segelbacher G (2016) Effects of forest fragmentation on the morphological and genetic structure of a dispersal-limited, endangered bird species. Nature Conservation, 16:39-58. https://doi.org/10.3897/natureconservation.16.10905\u003c/li\u003e\n \u003cli\u003eJollands N, Lermit J, Patterson M (2004) Aggregate eco-efficiency indices for New Zealand\u0026mdash;a principal components analysis. Journal of Environmental Management, 73(4):293-305. https://doi.org/10.1016/j.jenvman.2004.07.002\u003c/li\u003e\n \u003cli\u003eKlemann JRL, Vieira JS (2013) Assessing the extent of occurrence, area of occupancy, territory size, and population size of Marsh Tapaculo (\u003cem\u003eScytalopus iraiensis\u003c/em\u003e). Animal Biodiversity and Conservation, 36(1):47-57.\u003c/li\u003e\n \u003cli\u003eLima MC, Mariano EF (2019) Acoustic similarity among disjunct populations of \u003cem\u003eFormicarius colma\u003c/em\u003e Boddaert, 1783 (Aves: Formicariidae). Ci\u0026ecirc;ncia e Natura, Santa Maria, 41:1-7. https://doi.org/10.5902/2179460X33207\u003c/li\u003e\n \u003cli\u003eMata H (2005) Hist\u0026oacute;ria evolutiva das esp\u0026eacute;cies n\u0026atilde;o-andinas de \u003cem\u003eScytalopus\u0026nbsp;\u003c/em\u003einferida atrav\u0026eacute;s da variabilidade no DNA mitocondrial. Disserta\u0026ccedil;\u0026atilde;o, Pontif\u0026iacute;cia Universidade Cat\u0026oacute;lica do Rio Grande do Sul.\u003c/li\u003e\n \u003cli\u003eMata H, Fontana CS, Maur\u0026iacute;cio GN, Bornschein MR, Vasconcelos MF, Bonatto SL (2009) Molecular phylogeny and biogeography of the eastern Tapaculos (Aves: Rhinocryptidae: \u003cem\u003eScytalopus\u003c/em\u003e, \u003cem\u003eEleoscytalopus\u003c/em\u003e): Cryptic diversification in Brazilian Atlantic Forest. Molecular Phylogenetics and Evolution, 53(2):450-462. https://doi.org/10.1016/j.ympev.2009.07.017\u003c/li\u003e\n \u003cli\u003eMaur\u0026iacute;cio GN, Mata N, Bornschein MR, Cadena CD, Alvarenga H, Bonatto SL (2008) Hidden generic diversity in Neotropical birds: Molecular and anatomical data support a new genus for the \u003cem\u003e\u0026lsquo;\u0026lsquo;Scytalopus\u0026rdquo; indigoticus\u003c/em\u003e species-group (Aves: Rhinocryptidae). Molecular Phylogenetics and Evolution, 49(1):125-135. https://doi.org/10.1016/j.ympev.2008.06.017\u003c/li\u003e\n \u003cli\u003eOliveira VLC (2014) Influ\u0026ecirc;ncia do ru\u0026iacute;do ambiente em can\u0026ccedil;\u0026otilde;es de um p\u0026aacute;ssaro Oscine e um Suboscine da mesma popula\u0026ccedil;\u0026atilde;o. Disserta\u0026ccedil;\u0026atilde;o, Universidade Federal de Alfenas.\u003c/li\u003e\n \u003cli\u003ePacheco JF, Silveira LF, Aleixo A, Agne CE (2021) Annotated checklist of the birds of Brazil by the Brazilian Ornithological Records Committee\u0026mdash;second edition. Ornithology Research, 29:94-105. https://doi.org/10.1007/s43388-021-00058-x\u003c/li\u003e\n \u003cli\u003ePodos J, Warren PS (2007) The evolution of geographic variation in birdsong. Advances in the study of behavior or Elsevier, 37:403-458. https://doi.org/10.1016/S0065-3454(07)37009-5\u003c/li\u003e\n \u003cli\u003eR\u0026auml;s\u0026auml;nen K, Hendry AP (2008) Disentangling interactions between adaptive divergence and gene flow when ecology drives diversification. Ecology letters, 11(6):624-636. https://doi.org/10.1111/j.1461-0248.2008.01176.x\u003c/li\u003e\n \u003cli\u003eRepenning M (2017) Varia\u0026ccedil;\u0026atilde;o geogr\u0026aacute;fica em \u003cem\u003eSporophila\u003c/em\u003e (Aves: Thraupidae) com base em evid\u0026ecirc;ncias fenot\u0026iacute;picas, ecol\u0026oacute;gicas e gen\u0026eacute;ticas. Tese, Pontif\u0026iacute;cia Universidade Cat\u0026oacute;lica do Rio Grande do Sul.\u003c/li\u003e\n \u003cli\u003eSick H (1997) Ornitologia Brasileira. Nova Fronteira, Rio de Janeiro.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Atlantic Forest, Bioacoustics, Biogeography, Tyranni","lastPublishedDoi":"10.21203/rs.3.rs-6298732/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6298732/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study investigated the vocalizations of \u003cem\u003eScytalopus iraiensis\u003c/em\u003e with the \u0026nbsp;aim of identifying variations in vocalization between disjunct populations of \u0026nbsp;the species. We analyzed 30 sound recordings of the species obtained from the \u0026nbsp;Xeno-canto and Wiki Aves databases. The recordings were converted to the WAV \u0026nbsp;format and then played back using Audacity software to obtain the following \u0026nbsp;acoustic parameters: number of notes, duration, maximum frequency, and minimum \u0026nbsp;frequency. After obtaining the parameters, Principal Component Analysis (PCA) \u0026nbsp;was performed on the data in a smaller set of attributes. The PCA results were \u0026nbsp;used in a correlation analysis, in which one of the variables that showed a \u0026nbsp;high correlation (≥ 0.6) was excluded. After this process, the normality of \u0026nbsp;the variables was verified using the Shapiro-Wilk test (p \u0026lt; 0.001), and \u0026nbsp;those that did not meet the normality assumption were logarithmized. These \u0026nbsp;data were associated with the records of each region and the similarity \u0026nbsp;between the populations of the South and Southeast was assessed through a \u0026nbsp;Permutational Multivariate Analysis of Variance (PERMANOVA). Our results \u0026nbsp;showed that, despite the geographic isolation, the populations of \u003cem\u003eS. iraiensis\u003c/em\u003edid not present significant variation in their vocalizations (RF = 0.7279, p = 0.425), indicating that the time they were separated was not enough to cause changes in their songs.\u003c/p\u003e","manuscriptTitle":"Acoustic similarity between disjunct populations of the Marsh Tapaculo Scytalopus iraiensis Bornschein, Reinert \u0026amp; Pichorim, 1998 (Aves: Rhinocryptidae)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-09 12:08:11","doi":"10.21203/rs.3.rs-6298732/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"500f4db4-c6d6-4312-a7dc-692de9c61820","owner":[],"postedDate":"May 9th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-10-23T10:53:50+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-09 12:08:11","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6298732","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6298732","identity":"rs-6298732","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}