Daily dynamics of herbivores and natural enemies on an extrafloral nectary-bearing plant in a Neotropical wetland

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Here, we describe the composition and temporal dynamics of arthropod assemblages associated with Ipomoea carnea (Convolvulaceae), a widespread macrophyte with extrafloral nectaries in the Brazilian Pantanal. Over one year, we surveyed 30 plants monthly in three daily periods (morning, afternoon, night), documenting herbivores (specialists and generalists) and natural enemies (ants, wasps, and spiders). We recorded 1,996 herbivores and over 19,000 natural enemies, revealing clear temporal partitioning: ants were more abundant during the day and coincided with lower herbivore abundance, while spiders peaked at night and were associated with reduced occurrence of specialist herbivores. Wasps were less abundant overall but showed activity concentrated during daytime. These patterns suggest complementary roles of natural enemies across the daily cycle, resulting in continuous pressure on herbivores. Our findings highlight I. carnea as a natural platform to study arthropod community dynamics and provide a critical baseline for both ecological theory and conservation efforts in one of the world’s most threatened wetlands. temporal dynamics community ecology extrafloral nectaries herbivory Pantanal wetlands Figures Figure 1 Figure 2 Figure 3 Introduction Ecological interactions are highly variable because organisms are interconnected across multiple trophic levels (Del-Claro 2012). Among these, ant–plant mutualisms mediated by extrafloral nectaries (EFNs) are some of the most studied examples (Heil & McKey 2003 ; Rosumek et al. 2009 ; Rico-Gray & Oliveira 2007), yet antagonistic interactions such as herbivory and predation are equally central in regulating populations within ecosystems (Maron et al. 2002 ; Snyder et al. 2006 ). Together, these processes’ structure complex systems involving plants, herbivores, and natural enemies (Oliveira & Del-Claro 2005 ; Robinson et al. 2017 ). Extrafloral nectaries are widespread traits that attract ants, wasps, and spiders, providing indirect protection against herbivores while shaping arthropod community composition (Heil & McKey 2003 ; Lange et al. 2021; Alencar et al. 2023 ; Souza et al. 2024). Despite extensive research on EFN-mediated interactions, most studies have been restricted to daytime observations, overlooking potential diel variation in the activity of natural enemies and herbivores (Yamawo et al. 2014 ). Recent evidence suggests that temporal turnover in arthropod assemblages is an underexplored but fundamental aspect of community ecology, as natural enemies often display contrasting diel activity that may result in complementary effects on herbivore suppression (Oliveira & Del-Claro 2005 ; Snyder et al. 2006 ). Documenting such temporal dynamics is particularly relevant in wetlands, where environmental heterogeneity and seasonal hydrology strongly influence community assembly (Junk et al. 2006 ; Meurer et al. 2015 ; Nunes da Cunha et al. 2023 ). The Pantanal, the largest Neotropical wetland, harbors exceptional biodiversity (Pott et al. 2011a , b ; Dambros et al. 2018 ; Bulbol et al. 2023 ) but faces increasing threats from fire, droughts, and hydrological alteration (Tomas et al. 2019 ; Leal Filho et al. 2021 ; Marengo et al. 2021 ). In this context, EFN-bearing plants represent natural platforms to examine how daily cycles of activity mediate species interactions and how such interactions may be disrupted in threatened ecosystems. Ipomoea carnea (Convolvulaceae) is a perennial shrub common in Neotropical floodplains. It produces abundant EFNs and supports a diverse arthropod fauna, including both specialist and generalist herbivores, as well as multiple guilds of natural enemies (Keeler 1977 ; Frey 1995 ). Its widespread distribution and ecological role make it an ideal system to investigate community-level structuring of plant–arthropod interactions across diel cycles. Therefore, our study aimed to describe the daily dynamics of herbivores and natural enemies associated with I. carnea in the Pantanal. Specifically, we asked: (i) how do natural enemy guilds (ants, spiders, wasps) partition their activity across the day–night cycle? (ii) how does herbivore occurrence vary in response to enemy presence? and (iii) what community-level patterns emerge from diel turnover? By integrating descriptive observations with statistical models, our study provides a natural history baseline to generate hypotheses for future experimental tests and contributes to understanding how temporal complementarity in enemy activity sustains continuous defense in a highly dynamic wetland ecosystem.. Material and Methods Study area We chose a natural population of I. carnea located in Pantanal on Estrada Parque, Ponte 14 (19º34'37''S, 57º00'42''W) in the vicinity of the Base de Estudos do Pantanal of the Universidade Federal de Mato Grosso do Sul, Corumbá, Mato Grosso do Sul, Brazil. Ipomea carnea has extrafloral nectaries (EFNs) that attract ants and other potential predators of herbivorous insects, such as wasps and spiders (Keeler 1977 ; Steward and Keeler 1988 ). According to Keeler ( 1977 ), I. carnea has two EFN systems: one located at the distal part of the leaf petiole and the other composed of a ring of five nectaries at the base of each flower, around the pedicel, which are visited by ants of different species (Fig. 1 ). The Pantanal in Mato Grosso do Sul state, Brazil, is a large humid area formed by the coalescence of the Paraguay River and its tributaries. It has a subsumed tropical climate of the A type (Alvares et al. 2013 ). The vegetation of the sedimentary plain is a mosaic of aquatic areas, flooded fields, riparian forests, savannas (cerrado), cerradão, deciduous forest, and a large part of savannas and monodominant pioneer formations such as the cotton formed by species of the genus Ipomoea , popularly known as wild cotton (Pott et al. 2011b ). With accumulated precipitation data and the average monthly temperature of the sampled months, obtained from the CEMTEC-MS website (Centro de Monitoramento de Tempo, do Clima e dos Recursos Hídricos de Mato Grosso do Sul) for the Municipality of Corumbá, we constructed a climatic diagram to characterize the area during the months of study. According to the diagram, the region showed a dry and cold season (July–September), a wet and hot season (November–February), and a wet season with lower temperatures (March–July). Arthropod sampling Monthly collections were carried out from November 2012 to October 2013, in a population of I. carnea . Thirty individuals at a minimum height of 1.5 m from the ground and separated by a minimum distance of 5 m were chosen to cover the entire area for each monthly sampling. We recorded the number of herbivorous leaf insects and natural enemies (ants, wasps, spiders, and other arthropods) visiting extrafloral nectaries in three branches per individual. The specimens were identified by family and/or genus, and in some cases by species, using specialized literature (e.g.: Rafael et al. 2012 ; Baccaro et al. 2015 ). We also consulted specialists to confirm species identification (see acknowledgements). These records were performed in three periods: Daytime (8 a.m.–10 a.m.), afternoon (2 p.m.–4 p.m.), and nighttime (7 p.m. − 9 p.m.). On the other hand, herbivores were classified as specialists ( Charydotella rubicunda and Chellymorpha cribraria ) and generalists (the rest of the herbivores). Data analysis We used a descriptive and correlational approach to identify natural patterns in arthropod activity, which serves as a bias for generating hypotheses for future experimental studies. The occurrence (richness and abundance) of herbivores and natural enemies per plant (individual / plant) was organized by periods (morning, afternoon, and night). To verify the normality of the data, a Kolmogorov - Smirnov test was performed. One-way ANOVA was performed to compare the occurrence of herbivores and natural enemies between periods (daytime, afternoon, and night). To verify if the interaction of natural enemies with leaf herbivores, both in richness and abundance, is different among the period of the day, a Generalized Linear Mixed Model (GLMM) was used. To assemble this model, we verified the types of distribution in each analyzed set, using the package “ftdistrplus” (Delignette-Muller et al. 2010 ) with the Poisson Distribution being the most suitable for the database. The models (GLMM) and interactions were tested using the “glmer” function (Bates and Maechler 2010 ) considering each plant individual as a random factor. To verify whether the presence or absence of natural enemies is related to the occurrence of the main species of herbivores (specialists and generalists), we used the analysis of indicator species (Indval) considering the relative frequency of species and concentration of abundance in certain groups (absence and presence of natural enemies), using the package “Indicspecies” (De Cáceres 2013 ). These analyses were performed on the R platform (R Core Team 2021 ). Results Over one year of observations, we recorded 19,347 natural enemies and 1,996 herbivores associated with Ipomoea carnea . Ants were by far the most common enemies (71% of all records), followed by spiders (19%) and wasps (10%). Herbivores were more diverse, including a mix of generalists and two specialist taxa that were consistently observed feeding on leaves, often protected by frass or silk shelters. Temporal dynamics of natural enemies Natural enemy guilds exhibited clear diel turnover. Ants dominated the plants during morning and afternoon periods, reaching their lowest activity at night. Spiders, in contrast, were strongly nocturnal and represented the main predators after sunset. Wasps were less abundant overall, but their presence was concentrated during daylight hours (see full ANOVA outputs in Table S1a).. This temporal partitioning indicates that different groups of enemies alternate dominance over the daily cycle, maintaining continuous predation pressure on the plants (Fig. 2). Temporal dynamics of herbivores Herbivores also varied across the diel cycle, responding differently depending on their feeding strategy. Overall herbivore abundance and richness declined during the day when ant activity was highest. Specialist herbivores, which often display behavioral defenses against ants, became more frequent at night, when ants were less active. However, their abundance was lower in periods dominated by spiders, suggesting that nocturnal predation limits their activity. Generalist herbivores occurred throughout the cycle but tended to be less abundant during the afternoon, coinciding with higher wasp presence (model results in Table S1b). These patterns reveal that herbivore guilds respond differently to the temporal dynamics of their enemies. Enemy–herbivore associations Patterns of association between herbivores and natural enemies revealed guild-specific relationships. Ants were negatively related to both herbivore abundance and richness, particularly in daytime periods. Spiders were associated with reduced numbers of specialist herbivores at night, while wasps were negatively related to generalist herbivores during daylight (indicator species analysis in Table S1c). Together, these results support the hypothesis that complementary guilds of natural enemies exert time-specific control on herbivores (Table S1). Community-level structuring Indicator species analyses highlighted the strong temporal fidelity of both herbivores and their enemies. Specialist herbivores were disproportionately associated with nocturnal periods, whereas generalists occurred across all time windows. Ants were consistently linked to daylight, spiders to night, and wasps to afternoon activity. This turnover creates a “defense in shifts” dynamic in which different predator guilds replace one another over the diel cycle, resulting in nearly continuous top-down regulation of herbivores (Fig. 3). Discussion Our year-round observations of Ipomoea carnea in the Pantanal reveal clear daily patterns in the structure of associated arthropod communities. By surveying across morning, afternoon, and night, we documented that natural enemies partition their activity temporally, resulting in continuous pressure on herbivores. Ants were predominant during the day, spiders dominated at night, and wasps were most active during daylight. This temporal complementarity produced distinct associations with herbivores: ants coincided with reduced herbivore abundance and richness, spiders with lower occurrence of specialist herbivores at night, and wasps with modest but consistent reductions of generalist herbivores. These results confirm that different predator guilds contribute to herbivore control in complementary ways. Temporal structuring of natural enemies The temporal partitioning of ants, spiders, and wasps suggests that I. carnea benefits from a "defense in shifts", whereby different predator guilds exert pressure during different periods. Similar diel patterns have been observed in other systems, where ants dominate EFN interactions by day and spiders contribute at night (Oliveira & Del-Claro 2005 ; Yamawo et al. 2014 ; Lange et al. 2021). This complementarity may be driven by the differential preferences of these enemies for plant rewards: ants and spiders are often attracted by sugar-rich extrafloral nectar, while wasps can be rapidly recruited by volatile organic compounds released by plants (Röse et al. 2006 ; Pfannenstiel & Patt 2012 ; Alves-Silva et al. 2013 ). Thus, our findings expand current knowledge by documenting these complementary dynamics in a Neotropical wetland context. Herbivore responses and guild-specific associations Herbivores displayed guild-specific responses to enemy turnover. Specialists were most frequent at night, coinciding with reduced ant activity, but their numbers declined when spiders were abundant. This suggests that the defensive strategies of specialists, such as chemical camouflage or fecal shields, might be effective against ants and wasps but are circumvented by hunting modes of nocturnal spiders (Massuda & Trigo 2014 ). Generalists occurred throughout the day but tended to be less abundant in periods of high wasp activity. These guild-specific responses reinforce the idea that multiple enemy guilds act synergistically, creating a temporal mosaic of top-down effects (Snyder et al. 2006 ; Dáttilo et al. 2013). Contributions to natural history and wetland ecology Our findings highlight I. carnea as a focal species for understanding arthropod community dynamics in wetlands. EFNs serve as keystone resources that structure communities across trophic levels (Heil & McKey 2003 ; Rico-Gray & Oliveira 2007; Silva-Viana et al. 2021 ). The plant itself may actively mediate this temporal structuring; evidence from other systems shows that plants can modulate extrafloral nectar secretion, both in quantity and quality, across the diel cycle to attract specific defenders when most needed (Romero & Izzo 2004 ; Radhika et al. 2010 ; Falcão et al. 2014 ). By documenting how these interactions unfold across daily cycles, our study expands the scope of EFN ecology beyond diurnal observations. In the Pantanal, where seasonal flooding and habitat heterogeneity strongly influence plant–arthropod interactions (Junk et al. 2006 ; Tomas et al. 2019 ; Meurer et al. 2015 ; Yamazaki et al. 2016 ), such temporal structuring may be particularly important for maintaining ecological stability. Therefore, daily turnover should be considered an additional axis of variation shaping wetland arthropod communities. Limitations and future perspectives While our results demonstrate strong temporal patterns, they remain correlative. We did not manipulate EFN availability or exclude predator groups and therefore cannot infer direct causal effects of enemies on herbivores or plant fitness. Future experimental approaches, such as predator-exclusion assays, manipulations of nectar availability, or time-series analyses such as empirical dynamic modeling (Sugihara et al. 2012), could test the hypotheses generated here. Nevertheless, descriptive studies like ours are essential for identifying the natural history patterns that underpin experimental design and for capturing ecological dynamics that might otherwise remain unnoticed. We emphasize that such baselines are especially critical in ecosystems undergoing rapid environmental change. Final remarks By documenting daily turnover in arthropod communities associated with I. carnea , we provide rare evidence of temporal complementarity among natural enemies in a wetland ecosystem. These findings also carry applied relevance: diel interactions are likely sensitive to hydrological alterations and fire, two of the major emerging threats to the Pantanal (Tomas et al. 2019 ; Leal Filho et al. 2021 ; Arruda et al. 2022 ). Beyond their intrinsic ecological value, such baselines are critical for conservation, as they reveal how species interactions are structured in one of the world's most threatened wetlands. Our work thus offers both ecological insights and practical references for monitoring how intricate interaction networks respond to ongoing disturbances in the Pantanal. Declarations Ethical approval: For this type of study formal consent is not required. Funding: MOCN thanks to Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES by the fellowship and to Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP nº 2011/17708-0) by the project financing. REV thanks Fundação de Amparo à Pesquisa do Estado de Mato Grosso (FAPEMAT – nº 0602346/2017) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq - nº 313839/2019-0) to Desenvolvimento Científico Regional (DCR) support. Finally, REV thanks Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq – nº 302057/2023-4) for the Programa de Capacitação Institucional – PCI/MPEG support of the Ministério da Ciência, Tecnologia e Inovações (MCTI). Conflict of Interest : The authors declare that they have no conflict of interest. Author Contributions: MOCN and JRT contributed to the study conception and design. Material preparation and data collection were performed by MOCN. The first draft of the manuscript was written by MOCN, JRT and REV commented on previous versions of the manuscript. MOCN, JRT and REV read and approved of the final manuscript. Acknowledgments MOCN thanks to Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES by the fellowship and to Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP nº 2011/17708-0) by the project financing. To Professor Paulo Robson (UFMS), Douglas Araújo (UFMS), and to MSc. Andressa Figueiredo de Oliveira (UFMS) to ants, spider and herbivores identification, respectively, and Suzanne Koptur (Florida International University) for her suggestions in the previous version. REV thanks Fundação de Amparo à Pesquisa do Estado de Mato Grosso (FAPEMAT – nº 0602346/2017) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq - nº 313839/2019-0) to Desenvolvimento Científico Regional (DCR) support. Finally, REV thanks Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq – nº 302057/2023-4) for the Programa de Capacitação Institucional – PCI/INPP support of the Ministério da Ciência, Tecnologia e Inovações (MCTI). Availability of data and material: The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. References Alencar CLDS., Nogueira A, Vicente RE, Coutinho ÍAC (2023) Plant species with larger extrafloral nectaries produce better quality nectar when needed and interact with the best ant partners. Journal of Experimental Botany. http://dx.doi.org/10.1093/jxb/erad160 Alvares CA, Stape JL, Sentelhas PC, Gonçalves JDM, Sparovek G (2013) Köppen’s climate classification map for Brazil. Meteorologische Zeitschrift 22(6):711-728. Alves‐Silva E, Barônio GJ, Torezan‐Silingardi HM, Del‐Claro K. (2013) Foraging behavior of Brachygastra lecheguana (Hymenoptera: Vespidae) on Banisteriopsis malifolia (Malpighiaceae): Extrafloral nectar consumption and herbivore predation in a tending ant system. Entomological Science. https://doi.org/10.1111/ens.12004 Arruda FV, Teresa FB, Layme VM, Vicente RE, Camarota F, Izzo TJ (2022) Fire and flood: How the Pantanal ant communities respond to multiple disturbances?. Perspectives in Ecology and Conservation. http://dx.doi.org/10.1016/j.pecon.2022.04.002 Baccaro FB, Feitosa RM, Fernández F, Fernandes IO, Izzo TJ, Souza JLP, Solar RRC (2015) Guia para os gêneros de formigas do Brasil. Editora INPA, Manaus. https://doi.org/10.5281/zenodo.32912 Bates D, Maechler M (2010) lme4: Linear mixed-effects models using S4 classes. Available in: . Accessed on: Oct. 2019. Bulbol MM, Bartholomay PR, Oliveira ML, Somavilla A (2023) A new species of Olixon Cameron, 1887 (Hymenoptera: Rhopalosomatidae) and new records for the genus in Brazil. Zootaxa. http://dx.doi.org/10.11646/zootaxa.5277.3.9 Dambros J, França VV, Delabie JHC, Marques MI, Battirola LD (2018) Canopy ant assemblage (Hymenoptera: Formicidae) in two vegetation formations in the northern Brazilian Pantanal. Sociobiology 65(3): 358-369. https://doi.org/10.13102/sociobiology.v65i3.1932 Dattilo W, Rico‐Gray V, Rodrigues DJ, Izzo TJ (2013) Soil and vegetation features determine the nested pattern of ant–plant networks in a tropical rainforest. Ecological Entomology. https://doi.org/10.1111/een.12029 De Cáceres,M (2013) How to use the indicspecies package. Available in: . Accessed on: Oct. 2018. Delignette-Muller ML, Pouillot R, Denis JB, Dutang C (2010) Fitdistrplus: help to fit of a parametric distribution to non-censored or censored data. Available in: . Accessed on: Oct. 2019. Falcão JCF, Dáttilo W, Izzo TJ (2014) Temporal variation in extrafloral nectar secretion in different ontogenic stages of the fruits of Alibertia verrucosa S. Moore (Rubiaceae) in a Neotropical savanna. Journal of Plant Interactions. https://doi.org/10.1080/17429145.2013.782513 Frey R (1995) Ipomoea carnea ssp. fistulosa (Martius ex-Choisy) Austin: Taxonomy, biology and ecology reviewed and inquired. Tropical Ecology 36: 21-48. Heil M, McKey D (2003) Protective ant-plant interactions as model systems in ecological and evolutionary research. Annual Review of Ecology, Evolution, and Systematics. https://doi.org/10.1146/annurev.ecolsys.34.011802.132410 Junk WJ, Cunha CN, Wantzen KM, Petermann P, Strüssmann C, Marques MI, Adis J (2006) Biodiversity and its conservation in the Pantanal of Mato Grosso, Brazil. Aquatic Sciences. https://doi.org/10.1007/s00027-006-0851-4 Keeler KH (1977) The extrafloral nectaries of Ipomoea carnea (Convolvulaceae). American Journal of Botany 64: 1182-1188. Leal Filho W, Azeiteiro UM, Salvia AL, Fritzen B, Libonati R (2021) Fire in Paradise: Why the Pantanal is burning. Environmental Science and Policy. https://doi.org/10.1016/j.envsci.2021.05.005 Marengo JA, Cunha AP, Cuartas LA, Deusdará Leal KR, Broedel E, Seluchi ME, et al. (2021) Extreme drought in the Brazilian Pantanal in 2019–2020: characterization, causes, and impacts. Frontiers in Water. https://doi.org/10.3389/frwa.2021.639204 Maron JL, Combs JK, Louda SM (2002) Convergent demographic effects of insect attack on related thistles in coastal vs. continental dunes. Ecology. https://doi.org/10.1890/0012-9658(2002)083[3382:CDEOIA]2.0.CO;2 Massuda KF, Trigo JR (2014) Hiding in plain sight: Cuticular compound profile matching conceals a larval tortoise beetle in its host chemical cloud. Journal of chemical ecology. https://doi.org/10.1007/s10886-014-0424-2 Meurer E, Battirola LD, Delabie JHC, Marques MI (2015) Influence of the vegetation mosaic on ant (Formicidae: Hymenoptera) distributions in the Northern Brazilian Pantanal. Sociobiology. https://doi.org/10.13102/sociobiology.v62i3.359 Nunes da Cunha C, Bergier I, Tomas WM, Damasceno-Júnior GA, Santos SA, Assunção VA, et al. (2023) Classificação dos Macrohabitat do Pantanal Brasileiro: Atualização para Políticas Públicas e Manejo de Áreas Protegidas. Biodiversidade Brasileira (BioBrasil). https://doi.org/10.37002/biobrasil.v13i1.2223 Oliveira PS, Del-Claro K (2005) Multitrophic interactions in a neotropical savanna: ant-hemipteran systems, associated insect herbivores and a host plant. Biotic interactions in the tropics: their role in the maintenance of species diversity (eds. D.F.R.P. Burslem, M.A. Pinard and S.E. Hartley), pp. 414-438. Patt JM, Pfannenstiel RS (2008) Odor‐based recognition of nectar in cursorial spiders. Entomologia experimentalis et applicata 127(1): 64-71. Pfannenstiel RS, Patt JM (2012) Feeding on nectar and honeydew sugars improves survivorship of two nocturnal cursorial spiders. Biological Control. https://doi.org/10.1016/j.biocontrol.2012.07.013 Pott A, Oliveira AK, Damasceno-Junior GA, Silva JS (2011a) Plant diversity of the Pantanal wetland. Brazilian Journal of Biology. https://doi.org/10.1590/S1519-69842011000200005 Pott VJ, Pott A, Lima LCP, Moreira SN, Oliveira AKM (2011b) Aquatic macrophyte diversity of the Pantanal wetland and upper basin. Brazilian Journal of Biology. https://doi.org/10.1590/S1519-69842011000200004 R Core Team (2021) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Available in: . Accessed on: Oct. 2019. Radhika V, Kost C, Mithöfer A, Boland W (2010) Regulation of extrafloral nectar secretion by jasmonates in lima bean is light dependent. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.1009007107 Rafael JA, Melo GAR, Carvalho CJBD, Casari AS, Constantino R (2012) Insetos do Brasil: diversidade e taxonomia. Editora Holos, Ribeirão Preto. Robinson A, Inouye DW, Ogilvie JE, Mooney EH (2017) Multitrophic interactions mediate the effects of climate change on herbivore abundance. Oecologia. https://doi.org/10.1007/s00442-017-3934-0 Romero GQ, Izzo TJ (2004) Leaf damage induces ant recruitment in the Amazonian ant-plant Hirtella myrmecophila . Journal of tropical Ecology 675-682. https://doi.org/10.1017/S0266467404001749 Röse USR, Lewis J, Tumlinson JH (2006) Extrafloral nectar from cotton (Gossypium hirsutum) as a food source for parasitic wasps. Functional Ecology. https://doi.org/10.1111/j.1365-2435.2006.01071.x Rosumek FB, Silveira FA, Neves FDS, Barbosa NPDU, Diniz L, Oki Y, et al. (2009) Ants on plants: a meta-analysis of the role of ants as plant biotic defenses. Oecologia. https://doi.org/10.1007/s00442-009-1309-x Silva-Viana CB, Vicente RE, Kaminski LA, Izzo TJ (2021) Beyond the gardens: The extended mutualism from ant‐garden ants to nectary‐bearing plants growing in Amazon tree‐fall gaps. Biotropica. https://doi.org/10.1111/btp.12886 Snyder WE, Snyder GB, Finke DL, Straub CS (2006) Predator biodiversity strengthens herbivore suppression. Ecology letters. https://doi.org/10.1111/j.1461-0248.2006.00922.x Steward JL, Keeler KH (1988) Are there trade-offs among antiherbivore defenses in Ipomoea (Convolvulaceae)?. Oikos. https://doi.org/10.1007/BF00319409 Tomas WM, Oliveira Roque F, Morato RG, Medici PE, Chiaravalloti RM, Tortato FR, et al. (2019). Sustainability agenda for the Pantanal Wetland: perspectives on a collaborative interface for science, policy, and decision-making. Tropical Conservation Science. https://doi.org/10.1177/1940082919872634 Vicente RE, Dáttilo W, Izzo TJ (2014) Differential recruitment of Camponotus femoratus (Fabricius) ants in response to ant garden herbivory. Neotropical Entomology. https://doi.org/10.1007/s13744-014-0245-6 Yamawo A, Tagawa J, Hada Y, Suzuki N (2014) Different combinations of multiple defense traits in an extrafloral nectary‐bearing plant growing under various habitat conditions. Journal of Ecology. https://doi.org/10.1111/1365-2745.12169 Yamazaki L, Dambroz J, Meurer E, Vindica VF, Delabie JHC, Marques MI, Batirolla LD (2016) Ant community (Hymenoptera: Formicidae) associated with Callisthene fasciculata (Spr.) Mart. (Vochysiaceae) canopies in the Pantanal of Poconé, Mato Grosso, Brazil. Sociobiology. https://doi.org/10.13102/sociobiology.v63i2.824 Supplementary Files SupplementaryTableS1.docx Cite Share Download PDF Status: Published Journal Publication published 23 Feb, 2026 Read the published version in Wetlands → Version 1 posted Reviewers agreed at journal 28 Sep, 2025 Reviewers invited by journal 25 Sep, 2025 Editor invited by journal 24 Sep, 2025 Editor assigned by journal 24 Sep, 2025 First submitted to journal 23 Sep, 2025 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. 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Campinas","correspondingAuthor":false,"prefix":"","firstName":"José","middleName":"Roberto","lastName":"Trigo","suffix":""},{"id":520661805,"identity":"80b20d67-ef97-49ca-ac3e-03369e66ff89","order_by":2,"name":"Ricardo Eduardo Vicente","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA6UlEQVRIiWNgGAWjYDACZhDBBuNVgESYG0jRcgYkwkhACwOyFsY2MIlfi3k788PPFWU20Qa3Tyc+5p1XG83fDtTyo2IbTi0yh9mMJc+cS8vdcC53s+HMbcdzZxxmbGDsOXMbpxYJZh4Gyca2w7kbzvBuk/i47VhuA1ALM2MbXi3MP6Fatv9InHMsdz4RWtjgtjB8bKjJ3UBYC5uZZQPQLzPP8G6WnHHsQO5GoJaDeP3Cf/jxzYYym9y+M7wbP/PU1OXOO3/44IMfFbi1wIHCATB1GEweIKweCOQbwFQdUYpHwSgYBaNgZAEAPm9cbySMXI8AAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0003-2640-2537","institution":"Federal University of Amazonas: Universidade Federal do Amazonas","correspondingAuthor":true,"prefix":"","firstName":"Ricardo","middleName":"Eduardo","lastName":"Vicente","suffix":""}],"badges":[],"createdAt":"2025-09-21 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07:39:06","extension":"html","order_by":13,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":132091,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7666130/v1/076841c6f0d58ec4a70ca75f.html"},{"id":93014649,"identity":"18070792-f475-40ac-acb0-f8ac0f6011aa","added_by":"auto","created_at":"2025-10-08 07:39:05","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":2363689,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eIpomoea\u003c/em\u003e \u003cem\u003ecarnea\u003c/em\u003e in the wetland. Highlight in the presence of extrafloral nectaries at the base of leaves and flowers (EFN). Photos: Milton O. Cordova\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7666130/v1/41b9ba81cff05361c143b5a8.png"},{"id":93014656,"identity":"c7257b3a-48e9-4ce6-85ce-eb43e4c8afd3","added_by":"auto","created_at":"2025-10-08 07:39:06","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":2466529,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentatives of herbivores (A-D) and natural enemies (E-H) on \u003cem\u003eIpomoea carnea\u003c/em\u003e in the Brazilian Pantanal. A. \u003cem\u003eSchistocerca \u003c/em\u003esp. (Acrididae, Orthoptera), B. \u003cem\u003eDiabrotica\u003c/em\u003esp. (Chrysomelidae, Coleoptera), C. Tortricidae larvae (Lepidoptera), D. \u003cem\u003eChellymorpha cribaria \u003c/em\u003elarvae (Chrysomelidae, Coleoptera), E. \u003cem\u003eLeptogenys\u003c/em\u003e sp. (Formicidae, Hymenoptera), F. \u003cem\u003eCamponotus\u003c/em\u003e aff. \u003cem\u003emelanoticus \u003c/em\u003e(Formicidae, Hymenoptera)\u003cem\u003e, \u003c/em\u003eG. Araneidae (Arachnida), H. \u003cem\u003ePolistes\u003c/em\u003e aff. \u003cem\u003emajor \u003c/em\u003e(Vespidae, Hymenoptera). Photos: Milton O. Cordova\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7666130/v1/729fa202279fd16e301cd355.png"},{"id":93014652,"identity":"ac6e1b18-1c97-4405-9943-4c376679ec09","added_by":"auto","created_at":"2025-10-08 07:39:05","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":667992,"visible":true,"origin":"","legend":"\u003cp\u003eRole of period of the day in the behavior of leaf herbivores and natural enemies in the vegetative and reproductive stages of \u003cem\u003eIpomoea carnea\u003c/em\u003e in the Pantanal (Illustration: Milton O. Cordova)\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7666130/v1/ce56936901fcd340e4f5bf15.png"},{"id":103765713,"identity":"d4a6ac16-9685-4351-ba23-873adb1728b0","added_by":"auto","created_at":"2026-03-02 16:08:03","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":6603799,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7666130/v1/9a63620b-bbb6-4923-8276-c67adce281b0.pdf"},{"id":93014662,"identity":"ec276735-c036-478e-94ec-75bcd48f2de2","added_by":"auto","created_at":"2025-10-08 07:39:06","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":23904,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryTableS1.docx","url":"https://assets-eu.researchsquare.com/files/rs-7666130/v1/40ee5f6b9986f228fe0285f3.docx"}],"financialInterests":"","formattedTitle":"Daily dynamics of herbivores and natural enemies on an extrafloral nectary-bearing plant in a Neotropical wetland","fulltext":[{"header":"Introduction","content":"\u003cp\u003eEcological interactions are highly variable because organisms are interconnected across multiple trophic levels (Del-Claro 2012). Among these, ant\u0026ndash;plant mutualisms mediated by extrafloral nectaries (EFNs) are some of the most studied examples (Heil \u0026amp; McKey \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Rosumek et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Rico-Gray \u0026amp; Oliveira 2007), yet antagonistic interactions such as herbivory and predation are equally central in regulating populations within ecosystems (Maron et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Snyder et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Together, these processes\u0026rsquo; structure complex systems involving plants, herbivores, and natural enemies (Oliveira \u0026amp; Del-Claro \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Robinson et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eExtrafloral nectaries are widespread traits that attract ants, wasps, and spiders, providing indirect protection against herbivores while shaping arthropod community composition (Heil \u0026amp; McKey \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Lange et al. 2021; Alencar et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Souza et al. 2024). Despite extensive research on EFN-mediated interactions, most studies have been restricted to daytime observations, overlooking potential diel variation in the activity of natural enemies and herbivores (Yamawo et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Recent evidence suggests that temporal turnover in arthropod assemblages is an underexplored but fundamental aspect of community ecology, as natural enemies often display contrasting diel activity that may result in complementary effects on herbivore suppression (Oliveira \u0026amp; Del-Claro \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Snyder et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2006\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eDocumenting such temporal dynamics is particularly relevant in wetlands, where environmental heterogeneity and seasonal hydrology strongly influence community assembly (Junk et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Meurer et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Nunes da Cunha et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The Pantanal, the largest Neotropical wetland, harbors exceptional biodiversity (Pott et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2011a\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003eb\u003c/span\u003e; Dambros et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Bulbol et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) but faces increasing threats from fire, droughts, and hydrological alteration (Tomas et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Leal Filho et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Marengo et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In this context, EFN-bearing plants represent natural platforms to examine how daily cycles of activity mediate species interactions and how such interactions may be disrupted in threatened ecosystems.\u003c/p\u003e\u003cp\u003e\u003cem\u003eIpomoea carnea\u003c/em\u003e (Convolvulaceae) is a perennial shrub common in Neotropical floodplains. It produces abundant EFNs and supports a diverse arthropod fauna, including both specialist and generalist herbivores, as well as multiple guilds of natural enemies (Keeler \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1977\u003c/span\u003e; Frey \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1995\u003c/span\u003e). Its widespread distribution and ecological role make it an ideal system to investigate community-level structuring of plant\u0026ndash;arthropod interactions across diel cycles.\u003c/p\u003e\u003cp\u003eTherefore, our study aimed to describe the daily dynamics of herbivores and natural enemies associated with \u003cem\u003eI. carnea\u003c/em\u003e in the Pantanal. Specifically, we asked: (i) how do natural enemy guilds (ants, spiders, wasps) partition their activity across the day\u0026ndash;night cycle? (ii) how does herbivore occurrence vary in response to enemy presence? and (iii) what community-level patterns emerge from diel turnover? By integrating descriptive observations with statistical models, our study provides a natural history baseline to generate hypotheses for future experimental tests and contributes to understanding how temporal complementarity in enemy activity sustains continuous defense in a highly dynamic wetland ecosystem..\u003c/p\u003e"},{"header":"Material and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eStudy area\u003c/h2\u003e\u003cp\u003eWe chose a natural population of \u003cem\u003eI. carnea\u003c/em\u003e located in Pantanal on Estrada Parque, Ponte 14 (19\u0026ordm;34'37''S, 57\u0026ordm;00'42''W) in the vicinity of the Base de Estudos do Pantanal of the Universidade Federal de Mato Grosso do Sul, Corumb\u0026aacute;, Mato Grosso do Sul, Brazil. \u003cem\u003eIpomea carnea\u003c/em\u003e has extrafloral nectaries (EFNs) that attract ants and other potential predators of herbivorous insects, such as wasps and spiders (Keeler \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1977\u003c/span\u003e; Steward and Keeler \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e1988\u003c/span\u003e). According to Keeler (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1977\u003c/span\u003e), \u003cem\u003eI. carnea\u003c/em\u003e has two EFN systems: one located at the distal part of the leaf petiole and the other composed of a ring of five nectaries at the base of each flower, around the pedicel, which are visited by ants of different species (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The Pantanal in Mato Grosso do Sul state, Brazil, is a large humid area formed by the coalescence of the Paraguay River and its tributaries. It has a subsumed tropical climate of the A type (Alvares et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The vegetation of the sedimentary plain is a mosaic of aquatic areas, flooded fields, riparian forests, savannas (cerrado), cerrad\u0026atilde;o, deciduous forest, and a large part of savannas and monodominant pioneer formations such as the cotton formed by species of the genus \u003cem\u003eIpomoea\u003c/em\u003e, popularly known as wild cotton (Pott et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2011b\u003c/span\u003e). With accumulated precipitation data and the average monthly temperature of the sampled months, obtained from the CEMTEC-MS website (Centro de Monitoramento de Tempo, do Clima e dos Recursos H\u0026iacute;dricos de Mato Grosso do Sul) for the Municipality of Corumb\u0026aacute;, we constructed a climatic diagram to characterize the area during the months of study. According to the diagram, the region showed a dry and cold season (July\u0026ndash;September), a wet and hot season (November\u0026ndash;February), and a wet season with lower temperatures (March\u0026ndash;July).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eArthropod sampling\u003c/h3\u003e\n\u003cp\u003eMonthly collections were carried out from November 2012 to October 2013, in a population of \u003cem\u003eI. carnea\u003c/em\u003e. Thirty individuals at a minimum height of 1.5 m from the ground and separated by a minimum distance of 5 m were chosen to cover the entire area for each monthly sampling. We recorded the number of herbivorous leaf insects and natural enemies (ants, wasps, spiders, and other arthropods) visiting extrafloral nectaries in three branches per individual. The specimens were identified by family and/or genus, and in some cases by species, using specialized literature (e.g.: Rafael et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Baccaro et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). We also consulted specialists to confirm species identification (see acknowledgements). These records were performed in three periods: Daytime (8 a.m.\u0026ndash;10 a.m.), afternoon (2 p.m.\u0026ndash;4 p.m.), and nighttime (7 p.m. \u0026minus;\u0026thinsp;9 p.m.). On the other hand, herbivores were classified as specialists (\u003cem\u003eCharydotella rubicunda\u003c/em\u003e and \u003cem\u003eChellymorpha cribraria\u003c/em\u003e) and generalists (the rest of the herbivores).\u003c/p\u003e\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\u003ch2\u003eData analysis\u003c/h2\u003e\u003cp\u003eWe used a descriptive and correlational approach to identify natural patterns in arthropod activity, which serves as a bias for generating hypotheses for future experimental studies. The occurrence (richness and abundance) of herbivores and natural enemies per plant (individual / plant) was organized by periods (morning, afternoon, and night). To verify the normality of the data, a Kolmogorov - Smirnov test was performed. One-way ANOVA was performed to compare the occurrence of herbivores and natural enemies between periods (daytime, afternoon, and night). To verify if the interaction of natural enemies with leaf herbivores, both in richness and abundance, is different among the period of the day, a Generalized Linear Mixed Model (GLMM) was used. To assemble this model, we verified the types of distribution in each analyzed set, using the package \u0026ldquo;ftdistrplus\u0026rdquo; (Delignette-Muller et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) with the Poisson Distribution being the most suitable for the database. The models (GLMM) and interactions were tested using the \u0026ldquo;glmer\u0026rdquo; function (Bates and Maechler \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) considering each plant individual as a random factor. To verify whether the presence or absence of natural enemies is related to the occurrence of the main species of herbivores (specialists and generalists), we used the analysis of indicator species (Indval) considering the relative frequency of species and concentration of abundance in certain groups (absence and presence of natural enemies), using the package \u0026ldquo;Indicspecies\u0026rdquo; (De C\u0026aacute;ceres \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). These analyses were performed on the R platform (R Core Team \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eOver one year of observations, we recorded 19,347 natural enemies and 1,996 herbivores associated with \u003cem\u003eIpomoea carnea\u003c/em\u003e. Ants were by far the most common enemies (71% of all records), followed by spiders (19%) and wasps (10%). Herbivores were more diverse, including a mix of generalists and two specialist taxa that were consistently observed feeding on leaves, often protected by frass or silk shelters.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eTemporal dynamics of natural enemies\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eNatural enemy guilds exhibited clear diel turnover. Ants dominated the plants during morning and afternoon periods, reaching their lowest activity at night. Spiders, in contrast, were strongly nocturnal and represented the main predators after sunset. Wasps were less abundant overall, but their presence was concentrated during daylight hours (see full ANOVA outputs in Table S1a).. This temporal partitioning indicates that different groups of enemies alternate dominance over the daily cycle, maintaining continuous predation pressure on the plants (Fig. 2).\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eTemporal dynamics of herbivores\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eHerbivores also varied across the diel cycle, responding differently depending on their feeding strategy. Overall herbivore abundance and richness declined during the day when ant activity was highest. Specialist herbivores, which often display behavioral defenses against ants, became more frequent at night, when ants were less active. However, their abundance was lower in periods dominated by spiders, suggesting that nocturnal predation limits their activity. Generalist herbivores occurred throughout the cycle but tended to be less abundant during the afternoon, coinciding with higher wasp presence (model results in Table S1b). These patterns reveal that herbivore guilds respond differently to the temporal dynamics of their enemies.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eEnemy\u0026ndash;herbivore associations\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003ePatterns of association between herbivores and natural enemies revealed guild-specific relationships. Ants were negatively related to both herbivore abundance and richness, particularly in daytime periods. Spiders were associated with reduced numbers of specialist herbivores at night, while wasps were negatively related to generalist herbivores during daylight (indicator species analysis in Table S1c). Together, these results support the hypothesis that complementary guilds of natural enemies exert time-specific control on herbivores (Table S1).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eCommunity-level structuring\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eIndicator species analyses highlighted the strong temporal fidelity of both herbivores and their enemies. Specialist herbivores were disproportionately associated with nocturnal periods, whereas generalists occurred across all time windows. Ants were consistently linked to daylight, spiders to night, and wasps to afternoon activity. This turnover creates a \u0026ldquo;defense in shifts\u0026rdquo; dynamic in which different predator guilds replace one another over the diel cycle, resulting in nearly continuous top-down regulation of herbivores (Fig. 3).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eOur year-round observations of \u003cem\u003eIpomoea carnea\u003c/em\u003e in the Pantanal reveal clear daily patterns in the structure of associated arthropod communities. By surveying across morning, afternoon, and night, we documented that natural enemies partition their activity temporally, resulting in continuous pressure on herbivores. Ants were predominant during the day, spiders dominated at night, and wasps were most active during daylight. This temporal complementarity produced distinct associations with herbivores: ants coincided with reduced herbivore abundance and richness, spiders with lower occurrence of specialist herbivores at night, and wasps with modest but consistent reductions of generalist herbivores. These results confirm that different predator guilds contribute to herbivore control in complementary ways.\u003c/p\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eTemporal structuring of natural enemies\u003c/h2\u003e\u003cp\u003eThe temporal partitioning of ants, spiders, and wasps suggests that \u003cem\u003eI. carnea\u003c/em\u003e benefits from a \"defense in shifts\", whereby different predator guilds exert pressure during different periods. Similar diel patterns have been observed in other systems, where ants dominate EFN interactions by day and spiders contribute at night (Oliveira \u0026amp; Del-Claro \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Yamawo et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Lange et al. 2021). This complementarity may be driven by the differential preferences of these enemies for plant rewards: ants and spiders are often attracted by sugar-rich extrafloral nectar, while wasps can be rapidly recruited by volatile organic compounds released by plants (R\u0026ouml;se et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Pfannenstiel \u0026amp; Patt \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Alves-Silva et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Thus, our findings expand current knowledge by documenting these complementary dynamics in a Neotropical wetland context.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003eHerbivore responses and guild-specific associations\u003c/h2\u003e\u003cp\u003eHerbivores displayed guild-specific responses to enemy turnover. Specialists were most frequent at night, coinciding with reduced ant activity, but their numbers declined when spiders were abundant. This suggests that the defensive strategies of specialists, such as chemical camouflage or fecal shields, might be effective against ants and wasps but are circumvented by hunting modes of nocturnal spiders (Massuda \u0026amp; Trigo \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Generalists occurred throughout the day but tended to be less abundant in periods of high wasp activity. These guild-specific responses reinforce the idea that multiple enemy guilds act synergistically, creating a temporal mosaic of top-down effects (Snyder et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; D\u0026aacute;ttilo et al. 2013).\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\u003ch2\u003eContributions to natural history and wetland ecology\u003c/h2\u003e\u003cp\u003eOur findings highlight \u003cem\u003eI. carnea\u003c/em\u003e as a focal species for understanding arthropod community dynamics in wetlands. EFNs serve as keystone resources that structure communities across trophic levels (Heil \u0026amp; McKey \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Rico-Gray \u0026amp; Oliveira 2007; Silva-Viana et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The plant itself may actively mediate this temporal structuring; evidence from other systems shows that plants can modulate extrafloral nectar secretion, both in quantity and quality, across the diel cycle to attract specific defenders when most needed (Romero \u0026amp; Izzo \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Radhika et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Falc\u0026atilde;o et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). By documenting how these interactions unfold across daily cycles, our study expands the scope of EFN ecology beyond diurnal observations. In the Pantanal, where seasonal flooding and habitat heterogeneity strongly influence plant\u0026ndash;arthropod interactions (Junk et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Tomas et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Meurer et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Yamazaki et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), such temporal structuring may be particularly important for maintaining ecological stability. Therefore, daily turnover should be considered an additional axis of variation shaping wetland arthropod communities.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\u003ch2\u003eLimitations and future perspectives\u003c/h2\u003e\u003cp\u003eWhile our results demonstrate strong temporal patterns, they remain correlative. We did not manipulate EFN availability or exclude predator groups and therefore cannot infer direct causal effects of enemies on herbivores or plant fitness. Future experimental approaches, such as predator-exclusion assays, manipulations of nectar availability, or time-series analyses such as empirical dynamic modeling (Sugihara et al. 2012), could test the hypotheses generated here. Nevertheless, descriptive studies like ours are essential for identifying the natural history patterns that underpin experimental design and for capturing ecological dynamics that might otherwise remain unnoticed. We emphasize that such baselines are especially critical in ecosystems undergoing rapid environmental change.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\u003ch2\u003eFinal remarks\u003c/h2\u003e\u003cp\u003eBy documenting daily turnover in arthropod communities associated with \u003cem\u003eI. carnea\u003c/em\u003e, we provide rare evidence of temporal complementarity among natural enemies in a wetland ecosystem. These findings also carry applied relevance: diel interactions are likely sensitive to hydrological alterations and fire, two of the major emerging threats to the Pantanal (Tomas et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Leal Filho et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Arruda et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Beyond their intrinsic ecological value, such baselines are critical for conservation, as they reveal how species interactions are structured in one of the world's most threatened wetlands. Our work thus offers both ecological insights and practical references for monitoring how intricate interaction networks respond to ongoing disturbances in the Pantanal.\u003c/p\u003e\u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eEthical approval:\u003c/h2\u003e\u003cp\u003eFor this type of study formal consent is not required.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFunding:\u003c/h2\u003e\u003cp\u003eMOCN thanks to Coordena\u0026ccedil;\u0026atilde;o de Aperfei\u0026ccedil;oamento de Pessoal de N\u0026iacute;vel Superior - CAPES by the fellowship and to Funda\u0026ccedil;\u0026atilde;o de Amparo \u0026agrave; Pesquisa do Estado de S\u0026atilde;o Paulo (FAPESP n\u0026ordm; 2011/17708-0) by the project financing. REV thanks Funda\u0026ccedil;\u0026atilde;o de Amparo \u0026agrave; Pesquisa do Estado de Mato Grosso (FAPEMAT \u0026ndash; n\u0026ordm; 0602346/2017) and Conselho Nacional de Desenvolvimento Cient\u0026iacute;fico e Tecnol\u0026oacute;gico (CNPq - n\u0026ordm; 313839/2019-0) to Desenvolvimento Cient\u0026iacute;fico Regional (DCR) support. Finally, REV thanks Conselho Nacional de Desenvolvimento Cient\u0026iacute;fico e Tecnol\u0026oacute;gico (CNPq \u0026ndash; n\u0026ordm; 302057/2023-4) for the Programa de Capacita\u0026ccedil;\u0026atilde;o Institucional \u0026ndash; PCI/MPEG support of the Minist\u0026eacute;rio da Ci\u0026ecirc;ncia, Tecnologia e Inova\u0026ccedil;\u0026otilde;es (MCTI).\u003c/p\u003e\u003cp\u003e\u003cem\u003eConflict of Interest\u003c/em\u003e: The authors declare that they have no conflict of interest.\u003c/p\u003e\u003ch2\u003eAuthor Contributions:\u003c/h2\u003e\u003cp\u003eMOCN and JRT contributed to the study conception and design. Material preparation and data collection were performed by MOCN. The first draft of the manuscript was written by MOCN, JRT and REV commented on previous versions of the manuscript. MOCN, JRT and REV read and approved of the final manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e\u003cp\u003eMOCN thanks to Coordena\u0026ccedil;\u0026atilde;o de Aperfei\u0026ccedil;oamento de Pessoal de N\u0026iacute;vel Superior - CAPES by the fellowship and to Funda\u0026ccedil;\u0026atilde;o de Amparo \u0026agrave; Pesquisa do Estado de S\u0026atilde;o Paulo (FAPESP n\u0026ordm; 2011/17708-0) by the project financing. To Professor Paulo Robson (UFMS), Douglas Ara\u0026uacute;jo (UFMS), and to MSc. Andressa Figueiredo de Oliveira (UFMS) to ants, spider and herbivores identification, respectively, and Suzanne Koptur (Florida International University) for her suggestions in the previous version. REV thanks Funda\u0026ccedil;\u0026atilde;o de Amparo \u0026agrave; Pesquisa do Estado de Mato Grosso (FAPEMAT \u0026ndash; n\u0026ordm; 0602346/2017) and Conselho Nacional de Desenvolvimento Cient\u0026iacute;fico e Tecnol\u0026oacute;gico (CNPq - n\u0026ordm; 313839/2019-0) to Desenvolvimento Cient\u0026iacute;fico Regional (DCR) support. Finally, REV thanks Conselho Nacional de Desenvolvimento Cient\u0026iacute;fico e Tecnol\u0026oacute;gico (CNPq \u0026ndash; n\u0026ordm; 302057/2023-4) for the Programa de Capacita\u0026ccedil;\u0026atilde;o Institucional \u0026ndash; PCI/INPP support of the Minist\u0026eacute;rio da Ci\u0026ecirc;ncia, Tecnologia e Inova\u0026ccedil;\u0026otilde;es (MCTI).\u003c/p\u003e\u003ch2\u003eAvailability of data and material:\u003c/h2\u003e\u003cp\u003eThe datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAlencar CLDS., Nogueira A, Vicente RE, Coutinho \u0026Iacute;AC (2023) Plant species with larger extrafloral nectaries produce better quality nectar when needed and interact with the best ant partners. Journal of Experimental Botany. http://dx.doi.org/10.1093/jxb/erad160\u003c/li\u003e\n\u003cli\u003eAlvares CA, Stape JL, Sentelhas PC, Gon\u0026ccedil;alves JDM, Sparovek G (2013) K\u0026ouml;ppen\u0026rsquo;s climate classification map for Brazil. Meteorologische Zeitschrift 22(6):711-728.\u003c/li\u003e\n\u003cli\u003eAlves‐Silva E, Bar\u0026ocirc;nio GJ, Torezan‐Silingardi HM, Del‐Claro K. (2013) Foraging behavior of \u003cem\u003eBrachygastra lecheguana\u003c/em\u003e (Hymenoptera: Vespidae) on \u003cem\u003eBanisteriopsis malifolia\u003c/em\u003e (Malpighiaceae): Extrafloral nectar consumption and herbivore predation in a tending ant system. Entomological Science. https://doi.org/10.1111/ens.12004\u003c/li\u003e\n\u003cli\u003eArruda FV, Teresa FB, Layme VM, Vicente RE, Camarota F, Izzo TJ (2022) Fire and flood: How the Pantanal ant communities respond to multiple disturbances?. Perspectives in Ecology and Conservation. http://dx.doi.org/10.1016/j.pecon.2022.04.002\u003c/li\u003e\n\u003cli\u003eBaccaro FB, Feitosa RM, Fern\u0026aacute;ndez F, Fernandes IO, Izzo TJ, Souza JLP, Solar RRC (2015) Guia para os g\u0026ecirc;neros de formigas do Brasil. Editora INPA, Manaus. https://doi.org/10.5281/zenodo.32912\u003c/li\u003e\n\u003cli\u003eBates D, Maechler M (2010) lme4: Linear mixed-effects models using S4 classes. Available in: \u0026lt;http://CRAN.R-project.org/package=lme4\u0026gt;. Accessed on: Oct. 2019.\u003c/li\u003e\n\u003cli\u003eBulbol MM, Bartholomay PR, Oliveira ML, Somavilla A (2023) A new species of \u003cem\u003eOlixon\u003c/em\u003e Cameron, 1887 (Hymenoptera: Rhopalosomatidae) and new records for the genus in Brazil. Zootaxa. http://dx.doi.org/10.11646/zootaxa.5277.3.9\u003c/li\u003e\n\u003cli\u003eDambros J, Fran\u0026ccedil;a VV, Delabie JHC, Marques MI, Battirola LD (2018) Canopy ant assemblage (Hymenoptera: Formicidae) in two vegetation formations in the northern Brazilian Pantanal. Sociobiology 65(3): 358-369. https://doi.org/10.13102/sociobiology.v65i3.1932\u003c/li\u003e\n\u003cli\u003eDattilo W, Rico‐Gray V, Rodrigues DJ, Izzo TJ (2013) Soil and vegetation features determine the nested pattern of ant\u0026ndash;plant networks in a tropical rainforest. Ecological Entomology. https://doi.org/10.1111/een.12029\u003c/li\u003e\n\u003cli\u003eDe C\u0026aacute;ceres,M (2013) How to use the indicspecies package. Available in: \u0026lt;https://cran.r-project.org/web/packages/indicspecies/vignettes/indicspeciesTutorial.pdf\u0026gt;. Accessed on: Oct. 2018.\u003c/li\u003e\n\u003cli\u003eDelignette-Muller ML, Pouillot R, Denis JB, Dutang C (2010) Fitdistrplus: help to fit of a parametric distribution to non-censored or censored data. Available in: \u0026lt;http://cran.r-project.org/web/packages/fitdistrplus/index.html\u0026gt;. Accessed on: Oct. 2019.\u003c/li\u003e\n\u003cli\u003eFalc\u0026atilde;o JCF, D\u0026aacute;ttilo W, Izzo TJ (2014) Temporal variation in extrafloral nectar secretion in different ontogenic stages of the fruits of \u003cem\u003eAlibertia verrucosa\u003c/em\u003e S. Moore (Rubiaceae) in a Neotropical savanna. Journal of Plant Interactions. https://doi.org/10.1080/17429145.2013.782513\u003c/li\u003e\n\u003cli\u003eFrey R (1995) \u003cem\u003eIpomoea carnea\u003c/em\u003e ssp. \u003cem\u003efistulosa\u003c/em\u003e (Martius ex-Choisy) Austin: Taxonomy, biology and ecology reviewed and inquired. Tropical Ecology 36: 21-48.\u003c/li\u003e\n\u003cli\u003eHeil M, McKey D (2003) Protective ant-plant interactions as model systems in ecological and evolutionary research. Annual Review of Ecology, Evolution, and Systematics. https://doi.org/10.1146/annurev.ecolsys.34.011802.132410\u003c/li\u003e\n\u003cli\u003eJunk WJ, Cunha CN, Wantzen KM, Petermann P, Str\u0026uuml;ssmann C, Marques MI, Adis J (2006) Biodiversity and its conservation in the Pantanal of Mato Grosso, Brazil. Aquatic Sciences. https://doi.org/10.1007/s00027-006-0851-4\u003c/li\u003e\n\u003cli\u003eKeeler KH (1977) The extrafloral nectaries of \u003cem\u003eIpomoea carnea\u003c/em\u003e (Convolvulaceae). American Journal of Botany 64: 1182-1188.\u003c/li\u003e\n\u003cli\u003eLeal Filho W, Azeiteiro UM, Salvia AL, Fritzen B, Libonati R (2021) Fire in Paradise: Why the Pantanal is burning. Environmental Science and Policy. https://doi.org/10.1016/j.envsci.2021.05.005\u003c/li\u003e\n\u003cli\u003eMarengo JA, Cunha AP, Cuartas LA, Deusdar\u0026aacute; Leal KR, Broedel E, Seluchi ME, et al. (2021) Extreme drought in the Brazilian Pantanal in 2019\u0026ndash;2020: characterization, causes, and impacts. Frontiers in Water. https://doi.org/10.3389/frwa.2021.639204\u003c/li\u003e\n\u003cli\u003eMaron JL, Combs JK, Louda SM (2002) Convergent demographic effects of insect attack on related thistles in coastal vs. continental dunes. Ecology. https://doi.org/10.1890/0012-9658(2002)083[3382:CDEOIA]2.0.CO;2\u003c/li\u003e\n\u003cli\u003eMassuda KF, Trigo JR (2014) Hiding in plain sight: Cuticular compound profile matching conceals a larval tortoise beetle in its host chemical cloud. Journal of chemical ecology. https://doi.org/10.1007/s10886-014-0424-2\u003c/li\u003e\n\u003cli\u003eMeurer E, Battirola LD, Delabie JHC, Marques MI (2015) Influence of the vegetation mosaic on ant (Formicidae: Hymenoptera) distributions in the Northern Brazilian Pantanal. Sociobiology. https://doi.org/10.13102/sociobiology.v62i3.359\u003c/li\u003e\n\u003cli\u003eNunes da Cunha C, Bergier I, Tomas WM, Damasceno-J\u0026uacute;nior GA, Santos SA, Assun\u0026ccedil;\u0026atilde;o VA, et al. (2023) Classifica\u0026ccedil;\u0026atilde;o dos Macrohabitat do Pantanal Brasileiro: Atualiza\u0026ccedil;\u0026atilde;o para Pol\u0026iacute;ticas P\u0026uacute;blicas e Manejo de \u0026Aacute;reas Protegidas. Biodiversidade Brasileira (BioBrasil). https://doi.org/10.37002/biobrasil.v13i1.2223\u003c/li\u003e\n\u003cli\u003eOliveira PS, Del-Claro K (2005) Multitrophic interactions in a neotropical savanna: ant-hemipteran systems, associated insect herbivores and a host plant. Biotic interactions in the tropics: their role in the maintenance of species diversity (eds. D.F.R.P. Burslem, M.A. Pinard and S.E. Hartley), pp. 414-438. \u003c/li\u003e\n\u003cli\u003ePatt JM, Pfannenstiel RS (2008) Odor‐based recognition of nectar in cursorial spiders. Entomologia experimentalis et applicata 127(1): 64-71.\u003c/li\u003e\n\u003cli\u003ePfannenstiel RS, Patt JM (2012) Feeding on nectar and honeydew sugars improves survivorship of two nocturnal cursorial spiders. Biological Control. https://doi.org/10.1016/j.biocontrol.2012.07.013\u003c/li\u003e\n\u003cli\u003ePott A, Oliveira AK, Damasceno-Junior GA, Silva JS (2011a) Plant diversity of the Pantanal wetland. Brazilian Journal of Biology. https://doi.org/10.1590/S1519-69842011000200005\u003c/li\u003e\n\u003cli\u003ePott VJ, Pott A, Lima LCP, Moreira SN, Oliveira AKM (2011b) Aquatic macrophyte diversity of the Pantanal wetland and upper basin. Brazilian Journal of Biology. https://doi.org/10.1590/S1519-69842011000200004\u003c/li\u003e\n\u003cli\u003eR Core Team (2021) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Available in: \u0026lt;http://www.R-project.org/\u0026gt;. Accessed on: Oct. 2019.\u003c/li\u003e\n\u003cli\u003eRadhika V, Kost C, Mith\u0026ouml;fer A, Boland W (2010) Regulation of extrafloral nectar secretion by jasmonates in lima bean is light dependent. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.1009007107\u003c/li\u003e\n\u003cli\u003eRafael JA, Melo GAR, Carvalho CJBD, Casari AS, Constantino R (2012) Insetos do Brasil: diversidade e taxonomia. Editora Holos, Ribeir\u0026atilde;o Preto.\u003c/li\u003e\n\u003cli\u003eRobinson A, Inouye DW, Ogilvie JE, Mooney EH (2017) Multitrophic interactions mediate the effects of climate change on herbivore abundance. Oecologia. https://doi.org/10.1007/s00442-017-3934-0\u003c/li\u003e\n\u003cli\u003eRomero GQ, Izzo TJ (2004) Leaf damage induces ant recruitment in the Amazonian ant-plant \u003cem\u003eHirtella myrmecophila\u003c/em\u003e. Journal of tropical Ecology 675-682. https://doi.org/10.1017/S0266467404001749\u003c/li\u003e\n\u003cli\u003eR\u0026ouml;se USR, Lewis J, Tumlinson JH (2006) Extrafloral nectar from cotton (Gossypium hirsutum) as a food source for parasitic wasps. Functional Ecology. https://doi.org/10.1111/j.1365-2435.2006.01071.x\u003c/li\u003e\n\u003cli\u003eRosumek FB, Silveira FA, Neves FDS, Barbosa NPDU, Diniz L, Oki Y, et al. (2009) Ants on plants: a meta-analysis of the role of ants as plant biotic defenses. Oecologia. https://doi.org/10.1007/s00442-009-1309-x\u003c/li\u003e\n\u003cli\u003eSilva-Viana CB, Vicente RE, Kaminski LA, Izzo TJ (2021) Beyond the gardens: The extended mutualism from ant‐garden ants to nectary‐bearing plants growing in Amazon tree‐fall gaps. Biotropica. https://doi.org/10.1111/btp.12886\u003c/li\u003e\n\u003cli\u003eSnyder WE, Snyder GB, Finke DL, Straub CS (2006) Predator biodiversity strengthens herbivore suppression. Ecology letters. https://doi.org/10.1111/j.1461-0248.2006.00922.x\u003c/li\u003e\n\u003cli\u003eSteward JL, Keeler KH (1988) Are there trade-offs among antiherbivore defenses in \u003cem\u003eIpomoea\u003c/em\u003e (Convolvulaceae)?. Oikos. https://doi.org/10.1007/BF00319409\u003c/li\u003e\n\u003cli\u003eTomas WM, Oliveira Roque F, Morato RG, Medici PE, Chiaravalloti RM, Tortato FR, et al. (2019). Sustainability agenda for the Pantanal Wetland: perspectives on a collaborative interface for science, policy, and decision-making. Tropical Conservation Science. https://doi.org/10.1177/1940082919872634\u003c/li\u003e\n\u003cli\u003eVicente RE, D\u0026aacute;ttilo W, Izzo TJ (2014) Differential recruitment of \u003cem\u003eCamponotus femoratus\u003c/em\u003e (Fabricius) ants in response to ant garden herbivory. Neotropical Entomology. https://doi.org/10.1007/s13744-014-0245-6\u003c/li\u003e\n\u003cli\u003eYamawo A, Tagawa J, Hada Y, Suzuki N (2014) Different combinations of multiple defense traits in an extrafloral nectary‐bearing plant growing under various habitat conditions. Journal of Ecology. https://doi.org/10.1111/1365-2745.12169\u003c/li\u003e\n\u003cli\u003eYamazaki L, Dambroz J, Meurer E, Vindica VF, Delabie JHC, Marques MI, Batirolla LD (2016) Ant community (Hymenoptera: Formicidae) associated with \u003cem\u003eCallisthene fasciculata\u003c/em\u003e (Spr.) Mart. (Vochysiaceae) canopies in the Pantanal of Pocon\u0026eacute;, Mato Grosso, Brazil. Sociobiology. https://doi.org/10.13102/sociobiology.v63i2.824\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"wetlands","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"wela","sideBox":"Learn more about [Wetlands](https://www.springer.com/journal/13157)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/wela/default.aspx","title":"Wetlands","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"temporal dynamics, community ecology, extrafloral nectaries, herbivory, Pantanal, wetlands","lastPublishedDoi":"10.21203/rs.3.rs-7666130/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7666130/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eInteractions among plants, herbivores, and natural enemies are central to community ecology, yet most studies focus only on diurnal patterns, overlooking temporal variation across the day-night cycle. Here, we describe the composition and temporal dynamics of arthropod assemblages associated with \u003cem\u003eIpomoea carnea\u003c/em\u003e (Convolvulaceae), a widespread macrophyte with extrafloral nectaries in the Brazilian Pantanal. Over one year, we surveyed 30 plants monthly in three daily periods (morning, afternoon, night), documenting herbivores (specialists and generalists) and natural enemies (ants, wasps, and spiders). We recorded 1,996 herbivores and over 19,000 natural enemies, revealing clear temporal partitioning: ants were more abundant during the day and coincided with lower herbivore abundance, while spiders peaked at night and were associated with reduced occurrence of specialist herbivores. Wasps were less abundant overall but showed activity concentrated during daytime. These patterns suggest complementary roles of natural enemies across the daily cycle, resulting in continuous pressure on herbivores. Our findings highlight \u003cem\u003eI. carnea\u003c/em\u003e as a natural platform to study arthropod community dynamics and provide a critical baseline for both ecological theory and conservation efforts in one of the world\u0026rsquo;s most threatened wetlands.\u003c/p\u003e","manuscriptTitle":"Daily dynamics of herbivores and natural enemies on an extrafloral nectary-bearing plant in a Neotropical wetland","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-08 07:38:50","doi":"10.21203/rs.3.rs-7666130/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2025-09-28T22:04:16+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-25T15:10:50+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"Wetlands","date":"2025-09-24T15:39:45+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-24T10:37:04+00:00","index":"","fulltext":""},{"type":"submitted","content":"Wetlands","date":"2025-09-23T19:16:35+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"wetlands","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"wela","sideBox":"Learn more about [Wetlands](https://www.springer.com/journal/13157)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/wela/default.aspx","title":"Wetlands","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"5974b19a-ee2a-4d4d-b0c9-662aa7cf3ec9","owner":[],"postedDate":"October 8th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-03-02T16:04:41+00:00","versionOfRecord":{"articleIdentity":"rs-7666130","link":"https://doi.org/10.1007/s13157-026-02032-z","journal":{"identity":"wetlands","isVorOnly":false,"title":"Wetlands"},"publishedOn":"2026-02-23 15:58:31","publishedOnDateReadable":"February 23rd, 2026"},"versionCreatedAt":"2025-10-08 07:38:50","video":"","vorDoi":"10.1007/s13157-026-02032-z","vorDoiUrl":"https://doi.org/10.1007/s13157-026-02032-z","workflowStages":[]},"version":"v1","identity":"rs-7666130","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7666130","identity":"rs-7666130","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}