Stability of macroinvertebrate communities in vernal ponds following natural drying and refilling events

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Ward, Alice M. Belskis, Sara L. Hermann, Jon N. Sweetman This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6968044/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 20 Sep, 2025 Read the published version in Wetlands Ecology and Management → Version 1 posted 9 You are reading this latest preprint version Abstract Vernal ponds are vital components of forest ecosystems in the eastern United States, providing biodiversity support, water filtration, and flood regulation. Climate change may exacerbate hydrological fluctuations, altering the communities these seasonal wetlands support. This study examines the effects of drying disturbances on macroinvertebrate communities in vernal ponds, focusing on comparing biodiversity metrics before and after hydrological drawdown. We conducted weekly monitoring of pond inundation and macroinvertebrate sampling in five vernal ponds Central Pennsylvania during 2023. We measured alpha diversity using species richness and Shannon diversity, and calculated temporal beta diversity with Jaccard’s dissimilarity index, examining turnover and nestedness. We found no significant changes in alpha diversity metrics between pre- and post-drying periods. However, we observed a trend toward greater species loss (77% of dissimilarity) compared to gains (23%). Beta diversity patterns of turnover and nestedness were stable across temporal and spatial scales, suggesting that drying disturbances did not significantly affect community structure. These findings contrast with previous studies reporting significant shifts in community composition, potentially due to the adaptive strategies of macroinvertebrates. This research highlights the need for long-term studies to assess drying intensity and informs conservation strategies for vernal pond ecosystems in the context of climate change. variable inundation hydroperiod aquatic invertebrates biodiversity wetland ecology Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 INTRODUCTION Vernal ponds are isolated depressional wetlands that play a crucial role in forested ecosystems across the eastern United States. They provide several key ecosystem services, including natural filtration (Capps et al., 2014 ), important sites for faunal biodiversity (Colburn et al., 2008 ), and regulating flooding during heavy rainfall events (Leibowitz & Brooks, 2008 ). Vernal ponds experience a cyclic water regime where inundation often occurs in the spring following snowmelt and drawdown typically during the summer (Colburn, 2004 ). The period of inundation can vary between intermittent (e.g., days to weeks) to semipermanent (e.g., months; Cowardin et al., 1979 ). Vernal ponds are an important component of nutrient cycling within forested environments as a main contributor to biogeochemical processes, such as leaf-litter decomposition and denitrification (Capps et al., 2014 ). Additionally, vernal ponds provide critical habitats for amphibian reproduction and macroinvertebrate community assemblages due to the lack of established fish populations (Brown & Jung, 2005 ). Macroinvertebrates, such as aquatic insects, are important contributors to pond ecosystems through trophic-level interactions and cycling of nutrients. Macroinvertebrates in vernal ponds comprise multiple functional feeding groups, often serving as predators, prey, grazers, and recyclers of organic matter (Plenzler & Michaels, 2015 ). Pond hydroperiod is influential in affecting macroinvertebrate richness (Spencer et al., 1999 ), abundance (Leeper & Taylor, 1998a ), and reproductive success (Leeper & Taylor, 1998b ). While drawdown presents challenges for aquatic or semi-aquatic organisms, many taxa have evolved to encompass multiple adaptations to ensure survival during the this phase of the hydroperiod (Kenk, 1949 ; Wiggins et al., 1980 ). However, more frequent and extreme climatic events due to global climate change could lead vernal pools to become inundated or drawdown more frequently which could impact overall macroinvertebrate biodiversity. The fluctuating hydrological conditions of vernal ponds due to climate change could result in shifts in macroinvertebrate community composition, which can be assessed by examining changes in biodiversity metrics, including both alpha diversity (local scale species richness) and beta diversity (change in species between habitats). Diversity metrics can be sensitive measures of variation in communities at different spatial scales (e.g. α- versus β-diversity) but also over time (temporal β-diversity). Temporal β-diversity provides a metric for the changes in community composition within a habitat or ecosystem or across different locations, often considering changes from one specific time period to another (Legendre, 2019 ). Temporal beta diversity comprises two key components: (1) species turnover, which measures the turnover or loss/gain of species between timepoints and (2) nestedness, which is when species-poor communities are subsets of species-rich communities (Anderson et al., 2011 ; Baselga, 2010 ). Several recent studies have shown that partitioning beta diversity into turnover and nestedness components can provide additional insights into the factors that structure biodiversity patterns (Granath et al., 2024 ; Hill et al., 2017 ). These ecological measurements of communities can be powerful in deepening our understanding of the impact of environmental disturbances such as droughts or extreme rainfall, on macroinvertebrates in sensitive or temporary environments (Bae et al., 2014 ). Vernal ponds, which lack connections to permanent water sources, and usually dry substantially or completely during the summer months due to increases in temperature and evapotranspiration (Brooks, 2004 ). Since evapotranspiration is strongly temperature dependent, increasing temperatures are expected to exasperate/intensify drying rates and alter vernal pond hydrology and length of hydroperiod (Cartwright et al., 2022 ). This instability of pond hydroperiod has been shown to alter food web dynamics and food-chain lengths (Greig et al., 2013 ; Schriever & Williams, 2013 ), taxon succession (Devánová et al., 2023 ), and functional diversity (Coccia et al., 2024 ). Despite this, our understanding of how macroinvertebrate communities respond to drying in these pond ecosystems is limited. The goal of this paper is to investigate how macroinvertebrate biodiversity may differ between the pre-drying period and following refilling (post-drying) in vernal ponds. We predict that the drying acts as a disturbance that would prevent community reestablishment post-refilling. This research will provide valuable information on how climactically driven disturbances in vernal ponds might influence functionally important organisms, like macroinvertebrates. MATERIALS AND METHODS Study design We conducted a field survey to assess the impact of drying disturbances on macroinvertebrate communities in five vernal ponds located across three adjacent state forests in Central Pennsylvania: Bald Eagle State Forest (n = 2), Moshannon State Forest (n = 2), and Sproul State Forest (n = 1, Fig. 1 ). Initially, nine ponds were selected, but four ponds remained dry throughout the study period due to insufficient precipitation and were excluded from analysis. The drying event occurred mid-season and lasted approximately six weeks. Once ponds refilled, we immediately conducted sampling to capture macroinvertebrate community composition post-drying. Over a 10-week period from late May to the end of July 2023, we monitored these ponds weekly for water depth, pond length and width, and macroinvertebrate biodiversity. Only ponds that refilled after drying events were included in our analysis. Macroinvertebrate sampling Macroinvertebrates were sampled during periods of pond inundation both before drawdown and after refilling using a 500 µm D frame net from various pond microhabitats (e.g., emergent vegetation, substrate, limnetic zone, etc.; Batzer et al., 2005 ). A time-limited kick net sampling technique was employed to ensure consistency in sampling effort across ponds. Sampling time was standardized based on pond area, with a duration of 3 minutes for smaller ponds and up to 5 minutes for larger ponds (Nicolet et al., 2004 ). The collected samples were pooled to serve as a composite sample and stored in 80% ethanol (Batzer et al., 2005 ). All ethanol preserved specimens were identified to the lowest feasible taxonomic level, typically family or genus, and voucher specimens were sent to be verified by an independent taxonomist (Normandeau Associates, Stowe, PA, USA). Statistical methods All data analyses were performed in R version 4.2.3. To understand how pre and post drying events affected macroinvertebrate communities, we assessed macroinvertebrate diversity one week prior to (pre-drying) and one week following a drying event (post-drying). Alpha diversity was calculated as both species richness and Shannon diversity based on raw taxa counts using the vegan package (version 2.6-4, Oksanen et al., 2022 ). To test whether alpha diversity metrics changed between pre- and post-drying disturbance, we used a Wilcoxen-ranked sums test to account for a low sample size. Temporal beta diversity was calculated on a presence-absence matrix using multiple methods (Lawson et al., 2024 ). First, the Temporal Beta diversity Index (TBI) was calculated using the adespatial R package (version 0.3.22, Legendre, 2019 ). Jaccard’s dissimilarity index was used to quantify differences in species composition between pre- and post-drying disturbance. To account for changes in TBI values, we followed with a permutation test based on 9999 permutations and corrected with a Holm correction. TBI also returns the mean contribution of species losses and gains to the total dissimilarity, as described by Legendre (Legendre, 2019 ). Secondly, we applied the concept of partitioning beta diversity (here referred to as total beta diversity, i.e., Jaccard’s dissimilarity index) into species turnover and nestedness using the betapart package (version 1.6, Baselga & Orme, 2012 ). Spatial beta diversity was calculated as the mean of pairwise dissimilarities between ponds while temporal beta diversity is the pairwise dissimilarity between pre- and post-drying disturbance (Granath et al., 2024 ). We then compared spatial and temporal beta diversity for each component of beta diversity (e.g., turnover, nestedness, and total dissimilarity) using a Wilcoxon-ranked sums test. RESULTS A total of 507 macroinvertebrate individuals comprising 18 families were collected and identified from our study ponds (Table S1 ). Mean alpha diversity metrics did not change significantly following the observed drying disturbance compared to pre-drying (Richness: W = 20, p-value = 0.1425; Shannon Diversity: W = 19, p-value = 0.2101 ; Fig. 2 ). Across all sites sampled, there was a general trend of species losses post-drying on average than gains (Fig. 3 ). Averaged losses (0.773) accounted for more of the dissimilarity coefficient than gains (0.223) (i.e., species losses averaged 77% and gains averaged 23%; Fig. 3 ). One site located in Bald Eagle State Forest had a significant change (p < 0.05) in species prior to Holm correction (p-value = 0.0075; p-adjusted = 0.0375 ). Thus, there is weak evidence to suggest that the ponds showed changes in community composition between the two timepoints examined. To compare spatial and temporal beta diversity, we applied the concepts of turnover, nestedness, and total dissimilarity pre- and post-drying disturbance. Our results indicate there were no observed significant differences between either spatial or temporal beta diversity, regardless of the component examined. Specifically, for turnover, the comparison between pre- and post-drying disturbance to the temporal turnover index yielded test statistics that were not significant (pre-turnover: W = 5, p-value = 0.1425 ; post-turnover: W = 5, p-value = 0.1425; Fig. 4 ). Similarly, the comparison of the nestedness component between pre- and post-drying disturbance showed no significant differences when compared to the temporal metric (pre-nestedness: W = 10, p-value = 0.6905 ; post-nestedness: W = 11, p-value = 0.8413 ; Fig. 5 ). Lastly, total dissimilarity did not show any significance across the timepoints studied (pre-total: W = 16, p-value = 0.5476 ; post-total: W = 5, p-value = 0.1508 ; Fig. 6 ). The patterns of beta diversity remained consistent across both spatial and temporal aspects, suggesting that the drying disturbance did not significantly alter the macroinvertebrate community structure. DISCUSSION In this study, we investigated the response of macroinvertebrate communities in vernal ponds to drying disturbances, focusing on biodiversity metrics pre- and post-drying. We found that alpha diversity metrics remained relatively stable, despite observable fluctuations in species composition post-drying, indicating possible resilience to temporary drying disturbances. The resilience could be due to communities possessing adaptive strategies or preferences in habitat that mitigate the impacts of drying events (Kenk, 1949 ; Schiel & Buchwald, 2016 ; Wiggins et al., 1980 ). Our findings highlight the importance of understanding species-specific responses to environmental disturbances, as significant species losses following drying events imply potential shifts in community structure. Moreover, spatial and temporal beta diversity analyses revealed consistent turnover and nestedness patterns across sites and time, indicating stable community structure despite fluctuations in the hydroperiod. However, the short-term nature of our study and the limited number of sample sites may reduce the generalizability of our findings. Nevertheless, these findings highlight the ecological resilience of macroinvertebrate communities in unstable hydrological conditions. Effects of drying disturbances on macroinvertebrate biodiversity Despite our previous prediction indicating the drying disturbance would prevent macroinvertebrate reestablishment post-refilling, alpha diversity metrics were not significantly different between pre- and post-drying. However, while no significant differences were found in alpha diversity, a trend emerged indicating that species losses were more prevalent than species gains, with losses attributing 77% of the dissimilarity coefficient compared to 23% for gains. These losses were observed consistently across wetlands, although we did not identify the specific taxa lost or their functional traits, which could provide further insights into community responses to drying. Similarly, when examining beta diversity, our analysis found no significant differences pre- and post-drying between temporal (changes in species composition) and spatial (differences between ponds) beta diversity. The lack of observable differences suggests that the drying disturbance did not disrupt the patterns of species turnover and nestedness more than what would be expected from natural variability. The consistent patterns present across both temporal and spatial scales indicate that the drying event did not introduce significant changes in macroinvertebrate community structure. These findings contrast with previous research indicating significant shifts in macroinvertebrate communities post hydrological disruption. For instance, several studies found considerable alterations to macroinvertebrate community structure and trophic status following similar drying disturbances in temporary wetlands (Pérez-Bilbao et al., 2015 ; Schneider & Frost, 1996 ; Whiles et al., 1999 ; Williams, 1996 ). The lack of observable differences in diversity between pre- and post-drying in our study could be attributed to several factors. First, these communities may have evolved strategies to survive periodic disturbances, enabling them to persist despite environmental stress. Various macroinvertebrate groups, including trichopterans (Flint, 1958 ; Whiles et al., 1999 ) and coleopterans (Batzer & Wissinger, 1996 ), have developed strategies to migrate to more permanent habitats before drying. Other strategies could be attributed to the length of the life cycle where species with a short life span could be more likely to survive hydrological disruption than species with longer life cycles (Strachan et al., 2015 ). Site-specific characteristics, such as the presence of refugia, could have mitigated the effects of drying by providing saturated sediment or leaf litter (Batzer & Palik, 2007 ). Lastly, the duration and intensity of the drying disturbance may not have been severe enough to trigger substantial shifts in diversity. Both the timing and duration of drying have been found to be crucial in influencing the reestablishment of species post-refilling (Strachan et al., 2015 ); however, in the context of our research, these factors do not seem to have played a significant role in the observed changes in biodiversity. Additionally, it is important to note that a limitation of our study is the relatively low sample size due to the fact that four of our initial study sites did not rewet, which may have constrained our ability to detect subtle shifts in macroinvertebrate communities. Future research with larger sample sizes and increased replications would be beneficial to more robustly assess the impact of hydrological disturbances on macroinvertebrate diversity. Another possibility is that potential dispersal from adjacent permanent ponds might also have led to quick recovery. In our case, the presence of nearby ponds or more permanent water bodies could enhance the recovery of macroinvertebrate communities by providing a source of species that reintroduce individuals to disturbed ponds. The dispersal potential from adjacent ponds may thus increase the probability of species reestablishment and contribute to the observed stability in community structure. This potential for recovery through metacommunity dynamics could help explain why significant changes in diversity metrics were not detected, as the potential for more permanent systems may have mitigated more pronounced shifts in community composition. Our findings underscore the importance of understanding the implications of drying disturbances on macroinvertebrate biodiversity inhabiting vernal ponds. Although no significant changes in alpha and beta diversity were detected, the observed trend toward greater species losses highlight potential vulnerability of macroinvertebrate communities. Conversely, macroinvertebrate communities that inhabit vernal pond ecosystems may have evolved to survive in variably inundated environments and may be very resilient to alterations in hydroperiod. Moving forward, it is essential to conduct long-term studies and incorporate detailed assessments of drying intensity and site-specific variables to better understand the full implications of climate change on vernal pond ecosystems. In addition, considering temporal changes in species diversity following multiple refilling and drying events should be explored. Enhancing our knowledge in these areas will support the development of effective, holistic conservation strategies aimed at preserving the integrity and biodiversity of these crucial habitats in the face of ongoing climatic changes. Statements and Declarations The authors have no competing interests to declare that are relevant to the content of this article. ACKNOWLEDGEMENTS Author contributions: conceptualization : MSW, AMB, SLH, JNS;developing methods:MSW, JNS; conducting the research:MSW, JNS; data analysis: MSW, AMB, SLH, JNS; data interpretation: MSW, AMB, SLH, JNS; preparation figures & tables:MSW, AMB, SLH, JNS; writing: MSW, AMB, SLH, JNS. We thank undergraduate technicians C. Simonsen and T. Krivitski for critical contributions to fieldwork and processing laboratory samples and C. Smith of Normandeau Associates, Stowe, PA, USA for verification of macroinvertebrate samples. FUNDING This research was supported by the USDA National Institute of Food and Agriculture and McIntire Stennis appropriations under Project #PEN0 4787 and Accession #7003691. MSW is supported by the National Science Foundation Graduate Research Fellowship Program under Grant No. DGE1255832. SLH is supported by the Tombros Early Career Professorship and the Pests and Beneficial Species in Agricultural Production Systems (A1112) program, project award no. 2024-67013-42318, from the U.S. Department of Agriculture’s National Institute of Food and Agriculture. JNS is supported by the USDA National Institute of Food and Agriculture under Project #PEN04819 and Accession #7003691. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation. DATA AVAILABILITY Data and code are publicly available at (https://doi.org/10.26207/2x3h-5w21) and (https://github.com/masward/macro_dry.git) and upon request. This work was supported by the USDA National Institute of Food and Agriculture and McIntire-Stennis Appropriations under Project #PEN04787 and Accession #1027680. 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Cite Share Download PDF Status: Published Journal Publication published 20 Sep, 2025 Read the published version in Wetlands Ecology and Management → Version 1 posted Editorial decision: Revision requested 28 Jul, 2025 Reviews received at journal 28 Jul, 2025 Reviews received at journal 14 Jul, 2025 Reviewers agreed at journal 01 Jul, 2025 Reviewers agreed at journal 01 Jul, 2025 Reviewers invited by journal 26 Jun, 2025 Editor assigned by journal 26 Jun, 2025 Submission checks completed at journal 26 Jun, 2025 First submitted to journal 24 Jun, 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. 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-6968044","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":478430812,"identity":"785c6df2-6a83-4ec3-98e4-21224ec5521a","order_by":0,"name":"Mason S. Ward","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAuUlEQVRIiWNgGAWjYHAC9g8fKmzkIGw24rSwMc44k2ZMmhZmzrbDiQ1Ea9HtP2P2mOFMWvp2/jMGDB/KDhPWYnYjx9y4oMImd+eMHAPGGeeI0sK7QRrol9wNN3gMmHnbiNFy/uwGaaDKdIPzZwyY/xKl5UDuNpCWBIMDOQbMjERpuZH/2RDoMMOdM9IKDvacSyfGYccSHwCjUt6c//DGBz/KrAlrgQMDID5AgnqollEwCkbBKBgFWAEA3FlAz4w4zLwAAAAASUVORK5CYII=","orcid":"","institution":"The Pennsylvania State University","correspondingAuthor":true,"prefix":"","firstName":"Mason","middleName":"S.","lastName":"Ward","suffix":""},{"id":478430813,"identity":"72192b03-16d8-435a-8aab-0ad526ed5e56","order_by":1,"name":"Alice M. Belskis","email":"","orcid":"","institution":"The Pennsylvania State University","correspondingAuthor":false,"prefix":"","firstName":"Alice","middleName":"M.","lastName":"Belskis","suffix":""},{"id":478430818,"identity":"484c6fee-12e4-40b4-b7dc-5b8b5b656307","order_by":2,"name":"Sara L. Hermann","email":"","orcid":"","institution":"The Pennsylvania State University","correspondingAuthor":false,"prefix":"","firstName":"Sara","middleName":"L.","lastName":"Hermann","suffix":""},{"id":478430819,"identity":"2934101f-6c61-413e-9989-5409c5a39754","order_by":3,"name":"Jon N. Sweetman","email":"","orcid":"","institution":"The Pennsylvania State University","correspondingAuthor":false,"prefix":"","firstName":"Jon","middleName":"N.","lastName":"Sweetman","suffix":""}],"badges":[],"createdAt":"2025-06-24 17:23:17","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6968044/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6968044/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11273-025-10090-z","type":"published","date":"2025-09-20T15:57:37+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":85765725,"identity":"3170f2ea-f305-4adc-8015-2d62e048ab70","added_by":"auto","created_at":"2025-07-01 12:26:32","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":75432,"visible":true,"origin":"","legend":"\u003cp\u003eLocation of the ponds sampled in Bald Eagle State Forest (BDEG; n = 2), Moshannon State Forest (MSHN; n = 2), and Sproul State Forest (SPRL; n= 1). Each individual pond is represented by a black dot and a unique site identification label. Inset in the map is another map displaying the location of the three State Forests within the state of Pennsylvania, USA\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6968044/v1/1b8941f57f415c673bbab48a.png"},{"id":85765726,"identity":"7e78eae1-a972-4978-b513-3891e2044bdb","added_by":"auto","created_at":"2025-07-01 12:26:32","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":44290,"visible":true,"origin":"","legend":"\u003cp\u003eAlpha diversity, measured by species richness \u003cstrong\u003e(A)\u003c/strong\u003e and Shannon diversity \u003cstrong\u003e(B) \u003c/strong\u003ebetween the pre-drying and post-drying periods (pre-drying, \u003cem\u003en = 5\u003c/em\u003e; post-drying, \u003cem\u003en= 5\u003c/em\u003e). The boxplots display the median as a bolded horizontal line, the first and third quartiles, the minimum and maximum as “whiskers”, and the individual data points\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6968044/v1/acc43188603bab084d1989a8.png"},{"id":85765727,"identity":"538858c9-b991-49a0-ae4b-8d43272a9771","added_by":"auto","created_at":"2025-07-01 12:26:32","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":77293,"visible":true,"origin":"","legend":"\u003cp\u003eSpecies loss and gain plot for temporal beta diversity index (TBI). Colored squares represent where losses \u0026gt; gains, and the circle represents where gains \u0026gt; losses. The 1-1 line represents where species losses and gains would be equal with the area above denoting that species gains are greater than losses while the area below is where losses outweigh gains. Colors denote sites located within state forests (Bald Eagle = BDEG; Moshannon = MSHN; Sproul = SPRL). Size of points represent relative TBI values\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6968044/v1/ead09a88c3bdbfe213bb5e7d.png"},{"id":85766004,"identity":"3f7df734-2005-45fd-98be-16220d163e7a","added_by":"auto","created_at":"2025-07-01 12:34:32","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":26452,"visible":true,"origin":"","legend":"\u003cp\u003eSpatial and temporal turnover components of β-diversity for pre- and post-drying disturbance. Boxplots display the median as a bolded horizontal line, the first and third quartiles, the minimum and maximum as “whiskers”\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6968044/v1/7a1c086c1964b0d7703670ae.png"},{"id":85765730,"identity":"43287295-ba16-4257-89dd-4ddb7a94bac2","added_by":"auto","created_at":"2025-07-01 12:26:32","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":27485,"visible":true,"origin":"","legend":"\u003cp\u003eSpatial and temporal nestedness components of β-diversity for pre- and post-drying disturbance. Boxplots display the median as a bolded horizontal line, the first and third quartiles, the minimum and maximum as “whiskers”\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6968044/v1/5c327eb93d88761bdb5a598e.png"},{"id":85766005,"identity":"dc796e63-25f3-4c0f-ab1d-dc1576f4995a","added_by":"auto","created_at":"2025-07-01 12:34:32","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":25701,"visible":true,"origin":"","legend":"\u003cp\u003eSpatial and temporal total dissimilarity (β-diversity) with the median as a bolded horizontal line, the first and third quartiles, the minimum and maximum as “whiskers”\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-6968044/v1/9841a0e3a18bc19dc4fed6e0.png"},{"id":91889906,"identity":"7821e541-c317-49bf-b760-ba28265eed91","added_by":"auto","created_at":"2025-09-22 16:03:30","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":661286,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6968044/v1/b86cd6a5-af52-4d41-a9b7-ae2816c28eca.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Stability of macroinvertebrate communities in vernal ponds following natural drying and refilling events","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eVernal ponds are isolated depressional wetlands that play a crucial role in forested ecosystems across the eastern United States. They provide several key ecosystem services, including natural filtration (Capps et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2014\u003c/span\u003e), important sites for faunal biodiversity (Colburn et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2008\u003c/span\u003e), and regulating flooding during heavy rainfall events (Leibowitz \u0026amp; Brooks, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Vernal ponds experience a cyclic water regime where inundation often occurs in the spring following snowmelt and drawdown typically during the summer (Colburn, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). The period of inundation can vary between intermittent (e.g., days to weeks) to semipermanent (e.g., months; Cowardin et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e1979\u003c/span\u003e). Vernal ponds are an important component of nutrient cycling within forested environments as a main contributor to biogeochemical processes, such as leaf-litter decomposition and denitrification (Capps et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Additionally, vernal ponds provide critical habitats for amphibian reproduction and macroinvertebrate community assemblages due to the lack of established fish populations (Brown \u0026amp; Jung, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2005\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eMacroinvertebrates, such as aquatic insects, are important contributors to pond ecosystems through trophic-level interactions and cycling of nutrients. Macroinvertebrates in vernal ponds comprise multiple functional feeding groups, often serving as predators, prey, grazers, and recyclers of organic matter (Plenzler \u0026amp; Michaels, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Pond hydroperiod is influential in affecting macroinvertebrate richness (Spencer et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e1999\u003c/span\u003e), abundance (Leeper \u0026amp; Taylor, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e1998a\u003c/span\u003e), and reproductive success (Leeper \u0026amp; Taylor, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e1998b\u003c/span\u003e). While drawdown presents challenges for aquatic or semi-aquatic organisms, many taxa have evolved to encompass multiple adaptations to ensure survival during the this phase of the hydroperiod (Kenk, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1949\u003c/span\u003e; Wiggins et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e1980\u003c/span\u003e). However, more frequent and extreme climatic events due to global climate change could lead vernal pools to become inundated or drawdown more frequently which could impact overall macroinvertebrate biodiversity.\u003c/p\u003e \u003cp\u003eThe fluctuating hydrological conditions of vernal ponds due to climate change could result in shifts in macroinvertebrate community composition, which can be assessed by examining changes in biodiversity metrics, including both alpha diversity (local scale species richness) and beta diversity (change in species between habitats). Diversity metrics can be sensitive measures of variation in communities at different spatial scales (e.g. α- versus β-diversity) but also over time (temporal β-diversity). Temporal β-diversity provides a metric for the changes in community composition within a habitat or ecosystem or across different locations, often considering changes from one specific time period to another (Legendre, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Temporal beta diversity comprises two key components: (1) species turnover, which measures the turnover or loss/gain of species between timepoints and (2) nestedness, which is when species-poor communities are subsets of species-rich communities (Anderson et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Baselga, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Several recent studies have shown that partitioning beta diversity into turnover and nestedness components can provide additional insights into the factors that structure biodiversity patterns (Granath et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Hill et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). These ecological measurements of communities can be powerful in deepening our understanding of the impact of environmental disturbances such as droughts or extreme rainfall, on macroinvertebrates in sensitive or temporary environments (Bae et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eVernal ponds, which lack connections to permanent water sources, and usually dry substantially or completely during the summer months due to increases in temperature and evapotranspiration (Brooks, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Since evapotranspiration is strongly temperature dependent, increasing temperatures are expected to exasperate/intensify drying rates and alter vernal pond hydrology and length of hydroperiod (Cartwright et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). This instability of pond hydroperiod has been shown to alter food web dynamics and food-chain lengths (Greig et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Schriever \u0026amp; Williams, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), taxon succession (Dev\u0026aacute;nov\u0026aacute; et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), and functional diversity (Coccia et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Despite this, our understanding of how macroinvertebrate communities respond to drying in these pond ecosystems is limited. The goal of this paper is to investigate how macroinvertebrate biodiversity may differ between the pre-drying period and following refilling (post-drying) in vernal ponds. We predict that the drying acts as a disturbance that would prevent community reestablishment post-refilling. This research will provide valuable information on how climactically driven disturbances in vernal ponds might influence functionally important organisms, like macroinvertebrates.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy design\u003c/h2\u003e \u003cp\u003eWe conducted a field survey to assess the impact of drying disturbances on macroinvertebrate communities in five vernal ponds located across three adjacent state forests in Central Pennsylvania: Bald Eagle State Forest (n\u0026thinsp;=\u0026thinsp;2), Moshannon State Forest (n\u0026thinsp;=\u0026thinsp;2), and Sproul State Forest (n\u0026thinsp;=\u0026thinsp;1, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Initially, nine ponds were selected, but four ponds remained dry throughout the study period due to insufficient precipitation and were excluded from analysis. The drying event occurred mid-season and lasted approximately six weeks. Once ponds refilled, we immediately conducted sampling to capture macroinvertebrate community composition post-drying. Over a 10-week period from late May to the end of July 2023, we monitored these ponds weekly for water depth, pond length and width, and macroinvertebrate biodiversity. Only ponds that refilled after drying events were included in our analysis.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMacroinvertebrate sampling\u003c/h3\u003e\n\u003cp\u003eMacroinvertebrates were sampled during periods of pond inundation both before drawdown and after refilling using a 500 \u0026micro;m D frame net from various pond microhabitats (e.g., emergent vegetation, substrate, limnetic zone, etc.; Batzer et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). A time-limited kick net sampling technique was employed to ensure consistency in sampling effort across ponds. Sampling time was standardized based on pond area, with a duration of 3 minutes for smaller ponds and up to 5 minutes for larger ponds (Nicolet et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). The collected samples were pooled to serve as a composite sample and stored in 80% ethanol (Batzer et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). All ethanol preserved specimens were identified to the lowest feasible taxonomic level, typically family or genus, and voucher specimens were sent to be verified by an independent taxonomist (Normandeau Associates, Stowe, PA, USA).\u003c/p\u003e\n\u003ch3\u003eStatistical methods\u003c/h3\u003e\n\u003cp\u003eAll data analyses were performed in R version 4.2.3. To understand how pre and post drying events affected macroinvertebrate communities, we assessed macroinvertebrate diversity one week prior to (pre-drying) and one week following a drying event (post-drying). Alpha diversity was calculated as both species richness and Shannon diversity based on raw taxa counts using the \u003cem\u003evegan\u003c/em\u003e package (version 2.6-4, Oksanen et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). To test whether alpha diversity metrics changed between pre- and post-drying disturbance, we used a Wilcoxen-ranked sums test to account for a low sample size.\u003c/p\u003e \u003cp\u003eTemporal beta diversity was calculated on a presence-absence matrix using multiple methods (Lawson et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). First, the Temporal Beta diversity Index (TBI) was calculated using the \u003cem\u003eadespatial\u003c/em\u003e R package (version 0.3.22, Legendre, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Jaccard\u0026rsquo;s dissimilarity index was used to quantify differences in species composition between pre- and post-drying disturbance. To account for changes in TBI values, we followed with a permutation test based on 9999 permutations and corrected with a Holm correction. TBI also returns the mean contribution of species losses and gains to the total dissimilarity, as described by Legendre (Legendre, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSecondly, we applied the concept of partitioning beta diversity (here referred to as total beta diversity, i.e., Jaccard\u0026rsquo;s dissimilarity index) into species turnover and nestedness using the \u003cem\u003ebetapart\u003c/em\u003e package (version 1.6, Baselga \u0026amp; Orme, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Spatial beta diversity was calculated as the mean of pairwise dissimilarities between ponds while temporal beta diversity is the pairwise dissimilarity between pre- and post-drying disturbance (Granath et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). We then compared spatial and temporal beta diversity for each component of beta diversity (e.g., turnover, nestedness, and total dissimilarity) using a Wilcoxon-ranked sums test.\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cp\u003eA total of 507 macroinvertebrate individuals comprising 18 families were collected and identified from our study ponds (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). Mean alpha diversity metrics did not change significantly following the observed drying disturbance compared to pre-drying (Richness: \u003cem\u003eW\u0026thinsp;=\u0026thinsp;20, p-value\u0026thinsp;=\u0026thinsp;0.1425;\u003c/em\u003e Shannon Diversity: \u003cem\u003eW\u0026thinsp;=\u0026thinsp;19, p-value\u0026thinsp;=\u0026thinsp;0.2101\u003c/em\u003e; Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Across all sites sampled, there was a general trend of species losses post-drying on average than gains (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Averaged losses (0.773) accounted for more of the dissimilarity coefficient than gains (0.223) (i.e., species losses averaged 77% and gains averaged 23%; Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). One site located in Bald Eagle State Forest had a significant change (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in species prior to Holm correction \u003cem\u003e(p-value\u0026thinsp;=\u0026thinsp;0.0075; p-adjusted\u0026thinsp;=\u0026thinsp;0.0375\u003c/em\u003e). Thus, there is weak evidence to suggest that the ponds showed changes in community composition between the two timepoints examined.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eTo compare spatial and temporal beta diversity, we applied the concepts of turnover, nestedness, and total dissimilarity pre- and post-drying disturbance. Our results indicate there were no observed significant differences between either spatial or temporal beta diversity, regardless of the component examined. Specifically, for turnover, the comparison between pre- and post-drying disturbance to the temporal turnover index yielded test statistics that were not significant (pre-turnover: \u003cem\u003eW\u0026thinsp;=\u0026thinsp;5, p-value\u0026thinsp;=\u0026thinsp;0.1425\u003c/em\u003e; post-turnover: \u003cem\u003eW\u0026thinsp;=\u0026thinsp;5, p-value\u0026thinsp;=\u0026thinsp;0.1425;\u003c/em\u003e Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Similarly, the comparison of the nestedness component between pre- and post-drying disturbance showed no significant differences when compared to the temporal metric (pre-nestedness: \u003cem\u003eW\u0026thinsp;=\u0026thinsp;10, p-value\u0026thinsp;=\u0026thinsp;0.6905\u003c/em\u003e; post-nestedness: W\u0026thinsp;\u003cem\u003e=\u0026thinsp;11, p-value\u0026thinsp;=\u0026thinsp;0.8413\u003c/em\u003e; Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e). Lastly, total dissimilarity did not show any significance across the timepoints studied (pre-total: \u003cem\u003eW\u0026thinsp;=\u0026thinsp;16, p-value\u0026thinsp;=\u0026thinsp;0.5476\u003c/em\u003e; post-total: \u003cem\u003eW\u0026thinsp;=\u0026thinsp;5, p-value\u0026thinsp;=\u0026thinsp;0.1508\u003c/em\u003e; Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). The patterns of beta diversity remained consistent across both spatial and temporal aspects, suggesting that the drying disturbance did not significantly alter the macroinvertebrate community structure.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eIn this study, we investigated the response of macroinvertebrate communities in vernal ponds to drying disturbances, focusing on biodiversity metrics pre- and post-drying. We found that alpha diversity metrics remained relatively stable, despite observable fluctuations in species composition post-drying, indicating possible resilience to temporary drying disturbances. The resilience could be due to communities possessing adaptive strategies or preferences in habitat that mitigate the impacts of drying events (Kenk, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1949\u003c/span\u003e; Schiel \u0026amp; Buchwald, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Wiggins et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e1980\u003c/span\u003e). Our findings highlight the importance of understanding species-specific responses to environmental disturbances, as significant species losses following drying events imply potential shifts in community structure. Moreover, spatial and temporal beta diversity analyses revealed consistent turnover and nestedness patterns across sites and time, indicating stable community structure despite fluctuations in the hydroperiod. However, the short-term nature of our study and the limited number of sample sites may reduce the generalizability of our findings. Nevertheless, these findings highlight the ecological resilience of macroinvertebrate communities in unstable hydrological conditions.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eEffects of drying disturbances on macroinvertebrate biodiversity\u003c/h2\u003e \u003cp\u003eDespite our previous prediction indicating the drying disturbance would prevent macroinvertebrate reestablishment post-refilling, alpha diversity metrics were not significantly different between pre- and post-drying. However, while no significant differences were found in alpha diversity, a trend emerged indicating that species losses were more prevalent than species gains, with losses attributing 77% of the dissimilarity coefficient compared to 23% for gains. These losses were observed consistently across wetlands, although we did not identify the specific taxa lost or their functional traits, which could provide further insights into community responses to drying. Similarly, when examining beta diversity, our analysis found no significant differences pre- and post-drying between temporal (changes in species composition) and spatial (differences between ponds) beta diversity. The lack of observable differences suggests that the drying disturbance did not disrupt the patterns of species turnover and nestedness more than what would be expected from natural variability. The consistent patterns present across both temporal and spatial scales indicate that the drying event did not introduce significant changes in macroinvertebrate community structure.\u003c/p\u003e \u003cp\u003eThese findings contrast with previous research indicating significant shifts in macroinvertebrate communities post hydrological disruption. For instance, several studies found considerable alterations to macroinvertebrate community structure and trophic status following similar drying disturbances in temporary wetlands (P\u0026eacute;rez-Bilbao et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Schneider \u0026amp; Frost, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1996\u003c/span\u003e; Whiles et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Williams, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e1996\u003c/span\u003e). The lack of observable differences in diversity between pre- and post-drying in our study could be attributed to several factors. First, these communities may have evolved strategies to survive periodic disturbances, enabling them to persist despite environmental stress. Various macroinvertebrate groups, including trichopterans (Flint, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e1958\u003c/span\u003e; Whiles et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e1999\u003c/span\u003e) and coleopterans (Batzer \u0026amp; Wissinger, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1996\u003c/span\u003e), have developed strategies to migrate to more permanent habitats before drying. Other strategies could be attributed to the length of the life cycle where species with a short life span could be more likely to survive hydrological disruption than species with longer life cycles (Strachan et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Site-specific characteristics, such as the presence of refugia, could have mitigated the effects of drying by providing saturated sediment or leaf litter (Batzer \u0026amp; Palik, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Lastly, the duration and intensity of the drying disturbance may not have been severe enough to trigger substantial shifts in diversity. Both the timing and duration of drying have been found to be crucial in influencing the reestablishment of species post-refilling (Strachan et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2015\u003c/span\u003e); however, in the context of our research, these factors do not seem to have played a significant role in the observed changes in biodiversity. Additionally, it is important to note that a limitation of our study is the relatively low sample size due to the fact that four of our initial study sites did not rewet, which may have constrained our ability to detect subtle shifts in macroinvertebrate communities. Future research with larger sample sizes and increased replications would be beneficial to more robustly assess the impact of hydrological disturbances on macroinvertebrate diversity.\u003c/p\u003e \u003cp\u003eAnother possibility is that potential dispersal from adjacent permanent ponds might also have led to quick recovery. In our case, the presence of nearby ponds or more permanent water bodies could enhance the recovery of macroinvertebrate communities by providing a source of species that reintroduce individuals to disturbed ponds. The dispersal potential from adjacent ponds may thus increase the probability of species reestablishment and contribute to the observed stability in community structure. This potential for recovery through metacommunity dynamics could help explain why significant changes in diversity metrics were not detected, as the potential for more permanent systems may have mitigated more pronounced shifts in community composition.\u003c/p\u003e \u003cp\u003eOur findings underscore the importance of understanding the implications of drying disturbances on macroinvertebrate biodiversity inhabiting vernal ponds. Although no significant changes in alpha and beta diversity were detected, the observed trend toward greater species losses highlight potential vulnerability of macroinvertebrate communities. Conversely, macroinvertebrate communities that inhabit vernal pond ecosystems may have evolved to survive in variably inundated environments and may be very resilient to alterations in hydroperiod. Moving forward, it is essential to conduct long-term studies and incorporate detailed assessments of drying intensity and site-specific variables to better understand the full implications of climate change on vernal pond ecosystems. In addition, considering temporal changes in species diversity following multiple refilling and drying events should be explored. Enhancing our knowledge in these areas will support the development of effective, holistic conservation strategies aimed at preserving the integrity and biodiversity of these crucial habitats in the face of ongoing climatic changes.\u003c/p\u003e \u003c/div\u003e"},{"header":"Statements and Declarations","content":"\u003cp\u003eThe authors have no competing interests to declare that are relevant to the content of this article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eACKNOWLEDGEMENTS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions:\u0026nbsp;\u003c/strong\u003econceptualization\u003cstrong\u003e:\u0026nbsp;\u003c/strong\u003eMSW, AMB, SLH, JNS;developing methods:MSW, JNS; conducting the research:MSW, JNS; data analysis: MSW, AMB, SLH, JNS; data interpretation: MSW, AMB, SLH, JNS; preparation\u0026nbsp;figures \u0026amp; tables:MSW, AMB, SLH, JNS; writing: MSW, AMB, SLH, JNS.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWe thank undergraduate technicians C. Simonsen and T. Krivitski for critical contributions to fieldwork and processing laboratory samples and C. Smith of\u0026nbsp;Normandeau Associates, Stowe, PA, USA for verification of macroinvertebrate samples.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFUNDING\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was supported by the USDA National Institute of Food and Agriculture\u0026nbsp;and McIntire Stennis appropriations under Project #PEN0 4787 and Accession #7003691. MSW is supported by the National Science Foundation Graduate Research Fellowship Program under Grant No. DGE1255832. SLH is supported by the Tombros Early Career Professorship and the Pests and Beneficial Species in Agricultural Production Systems (A1112) program, project award no. 2024-67013-42318, from the U.S. Department of Agriculture\u0026rsquo;s National Institute of Food and Agriculture. JNS is supported by the\u0026nbsp;USDA National Institute of Food and Agriculture under Project #PEN04819 and Accession #7003691. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDATA AVAILABILITY\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData and code are publicly available at (https://doi.org/10.26207/2x3h-5w21) and (https://github.com/masward/macro_dry.git) and upon request. This work was supported by the USDA National Institute of Food and Agriculture and McIntire-Stennis Appropriations under Project #PEN04787 and Accession #1027680.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAnderson MJ, Crist TO, Chase JM, Vellend M, Inouye BD, Freestone AL, Sanders NJ, Cornell HV, Comita LS, Davies KF, Harrison SP, Kraft NJB, Stegen JC, Swenson NG (2011) Navigating the multiple meanings of \u0026beta; diversity: A roadmap for the practicing ecologist: Roadmap for beta diversity. Ecol Lett 14\u003cem\u003e:\u003c/em\u003e9\u0026ndash;28. https://doi.org/10.1111/j.1461-0248.2010.01552.x\u003c/li\u003e\n \u003cli\u003eBae M, Chon T, Park Y (2014) Characterizing differential responses of benthic macroinvertebrate communities to floods and droughts in three different stream types using a Self‐Organizing Map. 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J N Y Entomol Soc 66:59\u0026ndash;62.\u003c/li\u003e\n \u003cli\u003eGranath G, Hyseni C, Bini LM, Heino J, Ortega JCG, Johansson F (2024) Disentangling drivers of temporal changes in urban pond macroinvertebrate diversity. Urban Ecosyst 27:1027\u0026ndash;1039. https://doi.org/10.1007/s11252-023-01500-2\u003c/li\u003e\n \u003cli\u003eGreig HS, Wissinger SA, McIntosh AR (2013) Top‐down control of prey increases with drying disturbance in ponds: A consequence of non‐consumptive interactions\u003cem\u003e?\u0026nbsp;\u003c/em\u003eJ Anim Ecol 82:598\u0026ndash;607. https://doi.org/10.1111/1365-2656.12042\u003c/li\u003e\n \u003cli\u003eHill MJ, Heino J, Thornhill I, Ryves DB, Wood PJ (2017) Effects of dispersal mode on the environmental and spatial correlates of nestedness and species turnover in pond communities. 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Freshw Sci 32:964\u0026ndash;975.\u0026nbsp;https://doi.org/10.1899/13-008.1\u003c/li\u003e\n \u003cli\u003eSpencer M, Blaustein L, Schwartz SS, Cohen JE (1999) Species richness and the proportion of predatory animal species in temporary freshwater pools: Relationships with habitat size and permanence. Ecol Lett 2:157\u0026ndash;166.\u0026nbsp;https://doi.org/10.1046/j.1461-0248.1999.00062.x\u003c/li\u003e\n \u003cli\u003eStrachan SR, Chester ET, Robson BJ (2015) Freshwater invertebrate life history strategies for surviving desiccation. Springer Sci Rev 3:57\u0026ndash;75.\u0026nbsp;https://doi.org/10.1007/s40362-015-0031-9\u003c/li\u003e\n \u003cli\u003eWhiles MR, Goldowitz BS, Charlton RE (1999) Life history and production of a semi-terrestrial limnephilid caddisfly in an intermittent Platte River wetland. J N Am Benthol Soc 18:533\u0026ndash;544.\u0026nbsp;https://doi.org/10.2307/1468385\u003c/li\u003e\n \u003cli\u003eWiggins GB, Mackay RJ, Smith IM (1980) Evolutionary and ecological strategies of animals in annual temporary pools. Arch Hydrobiol Suppl 58:97\u0026ndash;206\u003c/li\u003e\n \u003cli\u003eWilliams DD (1996) Environmental constraints in temporary fresh waters and their consequences for the insect fauna. J N Am Benthol Soc 15:634\u0026ndash;650.\u0026nbsp;https://doi.org/10.2307/1467813\u003cem\u003e\u003cbr\u003e \u003c/em\u003e\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-ecology-and-management","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"wetl","sideBox":"Learn more about [Wetlands Ecology and Management](https://www.springer.com/journal/11273)","snPcode":"11273","submissionUrl":"https://submission.nature.com/new-submission/11273/3","title":"Wetlands Ecology and Management","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"variable inundation, hydroperiod, aquatic invertebrates, biodiversity, wetland ecology","lastPublishedDoi":"10.21203/rs.3.rs-6968044/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6968044/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eVernal ponds are vital components of forest ecosystems in the eastern United States, providing biodiversity support, water filtration, and flood regulation. Climate change may exacerbate hydrological fluctuations, altering the communities these seasonal wetlands support. This study examines the effects of drying disturbances on macroinvertebrate communities in vernal ponds, focusing on comparing biodiversity metrics before and after hydrological drawdown. We conducted weekly monitoring of pond inundation and macroinvertebrate sampling in five vernal ponds Central Pennsylvania during 2023. We measured alpha diversity using species richness and Shannon diversity, and calculated temporal beta diversity with Jaccard\u0026rsquo;s dissimilarity index, examining turnover and nestedness. We found no significant changes in alpha diversity metrics between pre- and post-drying periods. However, we observed a trend toward greater species loss (77% of dissimilarity) compared to gains (23%). Beta diversity patterns of turnover and nestedness were stable across temporal and spatial scales, suggesting that drying disturbances did not significantly affect community structure. These findings contrast with previous studies reporting significant shifts in community composition, potentially due to the adaptive strategies of macroinvertebrates. This research highlights the need for long-term studies to assess drying intensity and informs conservation strategies for vernal pond ecosystems in the context of climate change.\u003c/p\u003e","manuscriptTitle":"Stability of macroinvertebrate communities in vernal ponds following natural drying and refilling events","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-01 12:26:27","doi":"10.21203/rs.3.rs-6968044/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-07-28T18:03:37+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-07-28T17:21:50+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-07-14T15:43:03+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"337680908858472894227879622369386548059","date":"2025-07-01T15:04:11+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"331343023619219971544235394236873704174","date":"2025-07-01T13:47:38+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-06-26T13:40:28+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-06-26T13:37:18+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-06-26T13:31:26+00:00","index":"","fulltext":""},{"type":"submitted","content":"Wetlands Ecology and Management","date":"2025-06-24T17:21:31+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"wetlands-ecology-and-management","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"wetl","sideBox":"Learn more about [Wetlands Ecology and Management](https://www.springer.com/journal/11273)","snPcode":"11273","submissionUrl":"https://submission.nature.com/new-submission/11273/3","title":"Wetlands Ecology and Management","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"0551f522-3538-4f6d-b907-eca7993be03e","owner":[],"postedDate":"July 1st, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-09-22T16:02:18+00:00","versionOfRecord":{"articleIdentity":"rs-6968044","link":"https://doi.org/10.1007/s11273-025-10090-z","journal":{"identity":"wetlands-ecology-and-management","isVorOnly":false,"title":"Wetlands Ecology and Management"},"publishedOn":"2025-09-20 15:57:37","publishedOnDateReadable":"September 20th, 2025"},"versionCreatedAt":"2025-07-01 12:26:27","video":"","vorDoi":"10.1007/s11273-025-10090-z","vorDoiUrl":"https://doi.org/10.1007/s11273-025-10090-z","workflowStages":[]},"version":"v1","identity":"rs-6968044","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6968044","identity":"rs-6968044","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}