{"paper_id":"1df6b590-2e6a-4d4c-b1af-4cdc48b260f4","body_text":"Litter invertebrates display greater differences among locations than grass species in a temperate grassland | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Litter invertebrates display greater differences among locations than grass species in a temperate grassland Allison M. Wall, Philip S. Barton, Nick L. Schultz This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5486939/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 07 Apr, 2025 Read the published version in Journal of Insect Conservation → Version 1 posted 14 You are reading this latest preprint version Abstract Grasslands comprise a significant portion of terrestrial ecosystems, contributing an estimated 20% of global carbon stores. Biomass is recycled in these systems by photodegradation, biotic decomposition, and through disturbances such as fire or grazing. Yet the diversity of invertebrates and their potential role in biotic decomposition remains unclear in many grasslands worldwide. To help close this knowledge gap we conducted a litter bag experiment to test for the effects of four grass species (two native and two exotic) and two mesh sizes on invertebrate assemblages surveyed at three grassland sites in southeastern Australia. We collected 7,933 invertebrates across twelve arthropod orders and found that all sites had a diverse and abundant invertebrate community that readily interacted with detached grass litter. Study site had the largest effect on invertebrate composition, with significant differences found for Acari, Poduromorpha and Thysanoptera. Grass species identity also had a small but significant effect on invertebrate composition, but there was no effect of litter bag mesh size on the invertebrates. Conservation implications : We found that both geographic and floristic factors were important drivers of variation in grassland invertebrate communities linked to decomposing litter. Further research should focus on quantifying the role of invertebrates in grass decomposition rates and nutrient cycling to improve knowledge of grassland ecology. decomposition grassland ecology invertebrates litterbag Themeda triandra Poa labillardierei Figures Figure 1 Figure 2 Introduction Decomposition in grasslands is a crucial process contributing to soil formation and global carbon cycles (Hungate et al. 2017 ; Yang et al. 2019 ), with an estimated 34% of terrestrial carbon stored in grassland ecosystems (White et al. 2000 ). Up to 90% of terrestrial plant matter is decomposed rather than consumed in some ecosystems (Gessner et al. 2010 ). The factors that drive plant decomposition include climate variability (Wall et al. 2008 ), photodegradation (Austin & Vivanco 2006 ), nutrient availability (Bradford et al. 2016 ; Cuevas & Medina 1988 ), and the interacting effects of these factors (Aerts 1997 ; Garcia-Palacios et al. 2013 ; Makkonen et al. 2012 ). Invertebrates also play a key role in accelerating plant litter decomposition (Heděnec et al. 2022 ; Wall et al. 2008 ), but we know little about the local factors influencing invertebrate community composition in many grasslands (Gibson & New 2007 ). Invertebrates are an often-overlooked component of fauna due to their overwhelming diversity and lack of expertise among conservation practitioners (Cardoso et al. 2011 ). Invertebrate decomposers are a particularly important group due to their contribution to ecosystem function by facilitating biomass comminution, conversion and nutrient exchange (Handa et al. 2014 ; Hättenschwiler et al. 2005 ; Swift et al.1979). Invertebrates play an essential role in breaking down plant litter, dung, and carrion and returning nutrients to the soil and wider biosphere (Barton & Evans 2017 ; Bradford et al. 2016 ; Griffiths et al. 2021 ; Lavelle et al. 2006 ). Temperate grasslands are disproportionately rich in decomposer invertebrates and can support up to 100 tonnes per hectare of earthworms, microarthropods, fungi and bacteria (Orgiazzi et al. 2016 ), driving high rates of litter consumption (Heděnec et al. 2022 ). The stratum of leaf litter covering the topsoil is the initial habitat in which macro arthropods break down litter (Coleman et al. 2017 ). Tearing, shredding, ingesting and excreting mass results in weakened litter and smaller particles and a more labile state (Gongalsky 2021 ; Patoine et al. 2017 ). Acari and Collembola are considered the main litter transformers responsible for the break down and mixing of plant litter (Culliney 2013 ), with Millipedes, Isopods, and Insects also contributing. Detritivores produce enzymes to break down plant material, weakening and fragmenting plant litter with ingestion (Griffiths et al. 2021 ). These can include mites, millipedes and the larvae of Diptera and Coleoptera (Curry 1987 ), as well as up to 20,000 Collembola per square metre (Greenslade 2007 ). The movement of these arthropods as they forage and consume dead organic matter results in the subsoil becoming aerated and soil and nutrient mixing (Bagyaraj et al. 2016 ). Microarthropods most frequently inhabit the top 5cm of grassland ecosystems (Coleman et al. 2017 ). A loss of diversity in invertebrates and grass species has been shown to slow the cycling of carbon and nitrogen from leaf litter across several biomes (Handa et al. 2014 ) and in controlled laboratory experiments (Delgado-Baquerizo et al. 2020 ). The sensitivity of litter-dwelling arthropods has suggested they are a valuable measure of ecosystem health in temperate grasslands (Solascasas et al. 2022 ), with population fluxes rapidly responding to human management practices (Greenslade 2007 ). Despite the importance of arthropods in the initial breakdown of leaf litter, this is a rarely studied topic, and specific data on local scales is absent from many ecosystems (Hättenschwiler et al. 2005 ). Comparing invertebrate assemblages in native and exotic grass species can predict how ecosystems will respond to the invasion of alien grass species or be affected by degradation. Belnap & Phillips ( 2001 ) showed how the invasion of an exotic grass resulted in lower species richness and overall invertebrate abundance. A similar decline in local insect diversity was recorded by Litt and Steidl ( 2010 ) in Eragrostis lehmanniana -invaded native pastures in Arizona, though a few orders favoured the change. Similar trends in response to invasive species have been noted in South Africa (Samways et al. 1996 ), and South Australia (Clay 2014 ) as well as the increased susceptibility of specialist invertebrate species compared to generalists (Yoshioka et al. 2010 ). In Australian temperate grasslands, Miller and New ( 1997 ) explored the change in ant assemblages in intact and degraded sites and found that while the composition of the ant community varied, the composition of functional groups did not (Miller & New 1997 ). Little research has been published on grassland invertebrates in southern Australia, and no systematic study has been undertaken on Victoria’s grasslands (Yen 1999 ). This knowledge gap concerns ecologists as 70% of insect endemism is estimated in Australia (Chapman 2009 ), and less than half of the insect fauna is described (Saunders et al. 2021 ; Zborowski & Storey 2017 ). Further evidence is needed on the role of invertebrates in decomposition and how the nutrient status of grass species mediates this (Delgado-Baquerizo et al. 2020 ). We cannot effectively manage our current remnants without understanding the influence of invertebrates, and how transitions towards a more alien assemblage of plants might impact the efficacy of plant breakdown and associated nutrient cycling. Furthermore, knowledge gaps remain regarding the identity of the invertebrates that break down litter (Rosenberg et al. 2023 ). This shortfall has management repercussions, making us unable to predict the impact of exotic plant invasions, exotic invertebrates, and climate change (Decaëns 2010 ). The range of natural grasslands in southern Australia is heavily depleted, with < 5% of the once extensive ecosystem now considered intact (Roy & Delpratt 2006 ). Many grasslands in Australia now feature naturalised, self-sustaining populations of exotic grass (Morgan 1998 ; New 2019 ), which has several implications for grasslands’ function and ecological processes, including decomposition. The native grasses of Australia are adapted to low-nutrient and shallow soils (Fay et al. 2015 ; King & Hutchinson 1983 ; Orians & Milewski 2007 ). The grass decomposition trajectories may differ from those of higher-nutrient systems, with the photodegradation of high-lignin grasses acting as a primer for further decomposition (Butler et al. 2023 ). Nevertheless, the influence of leaf litter types on the local invertebrate decomposer community has not been tested. Aims We aimed to determine which invertebrate orders are present in the initial breakdown of detached grass litter. Within this context, we asked: Do invertebrate orders exhibit a preference for particular grass species? Do invertebrates respond differently to native and exotic grass species? Does the mesh aperture exclude elements of the invertebrates present? Does the composition of invertebrates vary between grassland locations? Methods Our research took place in the temperate grasslands of southeast Australia, approximately 50 km northwest of Melbourne. We used three grassland sites within a 15 km radius. The sites are named by location, and all are temperate grassland, varying in vegetation structure, aspect and management. Ingliston was located on a 25-metre roadside strip of remnant grassland that consisted of short Themeda triandra amongst an array of forbs and herbs. Myrniong was a grazing exclusion plot with dense swards of Themeda triandra. Parwan was a low rainfall plains grassland site on private property that receives ecological burns to reduce biomass, with tussocks of Austrostipa, Rytidosperma, Themeda and other grasses and forbs. Our experiment involved deploying 15 x 15 cm mesh bags (constructed sensu Butler et al. 2023 ; see Supplementary Figure S1 ) containing different grass species to aid in sampling the invertebrate community that colonises the detached grass litter. Mesh bags provide a means to observe natural processes such as litter breakdown and decomposition (Kitz et al. 2015 ; Schädler & Brandl 2005 ; Wall et al. 2008 ), and the aperture size of the bags determines the size classes of invertebrates that can access the litter they contain (Knops et al. 2001 ; Milcu & Manning 2011 ). Hitherto, grassland studies with litterbags have focussed on the decomposition of litter. This study is the first recorded instance (to the author’s knowledge) of using litter bags to sample invertebrates present. We chose four grass species, two native to the grassland study sites ( Themeda triandra Forssk., Poa labillardierei Steud. ) and two locally abundant introduced and naturalised species ( Dactylis glomerata L., Holcus lanatus L. ) . We harvested the grass near the Myrniong and Ingliston sites. We used secateurs to cut blades approximately five centimetres from the ground’s surface. We then removed the culms to a practicable degree before bagging samples so that most collected samples comprised leaf blades. We then refrigerated the grass collections at 4ºC until needed. We prepared each mesh bag for deployment by adding 10 g of air-dried grass to each bag. We added a plastic ‘pizza table’ to each bag to maintain airflow and prevent compaction, and we stapled the bags shut. We deployed 24 grass-filled mesh bags in each grassland on 9th November 2022, including six replicates of each of the four species per site. Half of the bags for each species used small-aperture mesh (2 mm) and half used large-aperture mesh (5 mm). When deploying each bag we trimmed the existing growth to ground-level to allow contact of the bag with the soil surface. Each bag was secured to the ground with a metal pin. A sturdy mesh cage was secured over groups of four bags to prevent disturbance from animals (see Supplementary Figure S1 ). We collected the bags on 8th December 2022, a month after deployment, using a small serving board under each bag to prevent loss of invertebrates when transferring litter bags to a paper bag. We used a Tullgren-Funnel apparatus in a glasshouse at Federation University, Mount Helen, to extract invertebrates from the grass samples (See Supplementary Figure S1 ). We placed the litterbags atop a zinc plate on top of the funnels. Invertebrates fell through the zinc plate as they moved away from light and were collected in a specimen container with 30 mL ethanol affixed at the bottom of the funnel. We allowed five days for this process to occur, with the alcohol levels checked after two days. We then removed the specimen jars from the funnels and stored them upright with complete lids. We used a Nikon SMZ 745 stereomicroscope with a 10–40 x magnification range for identification. We then transferred each sample to a Petri dish via pipette and tallied individuals along the sections of a lined grid. A clicker counter was used for larger samples. We recounted the first five samples at the end to ensure methodology had not evolved, but no significant differences were noted. Specimens were identified to order using Zborowski and Storey ( 2017 ), Daley and Ellingsen ( 2012 ) and Gooderham and Tsyrlin ( 2002 ). Data was compiled in a Microsoft Excel spreadsheet. See Supplementary Data 2 for raw data. All data analyses and figure production were conducted in R (R Core Team 2023 ) except where stated. We constructed non-metric multidimensional scaling (nMDS) ordination diagrams using the vegan package (Oksanen et al. 2019 ) based on a square-root transformation of the ordinal abundance data and a Bray-Curtis similarity matrix. We used ellipses projected onto the nMDS plots that grouped sites according to the sample variables (grass species, grass origin, mesh size and site) to observe how they influenced invertebrate composition. Using the statistical software PRIMER (Clarke & Gorley 2013 ), we tested for the influence of sample variables on invertebrate composition using analysis of similarity (ANOSIM) and similarity percentage (SIMPER) partitions (Clarke 1993 ). PRIMER was also used to calculate the following univariate diversity indices for each site: Pielou’s evenness, Shannon and Simpson’s diversity indices, and Margalef richness. Box-and-whisker plots were constructed using the package ggpubr (Kassambara 2023 ) to visualise the differences in diversity indices among the sites. Non-parametric Kruskal-Wallis tests were used to test for differences in diversity indices among sites, as the data failed the Shapiro-Wilk normality tests. Results We sampled a total of 7,933 individual invertebrates from the three sites. These invertebrates belonged to twelve different orders of insects and insect allies (Table 1). The number of captured invertebrates varied from 15 to 535 individuals per litter bag, with a mean of 110 ± 10.1. The overall abundance of invertebrates differed significantly among locations (Fig. 2 a), but the ordinal richness was similar among the grassland sites, with the sample means of 6.6 ± 0.3 for Ingliston, 7.8 ± 0.4 for Myrniong, and 6.5 ± 0.3 for Parwan. Evenness among sites varied significantly and was highest at Myrniong (Fig. 2 c). We excluded the larvae present from further analysis. The most abundant groups collected were Poduromorpha, Acari and Thysanoptera, with a combination of these three groups being the most abundant at each location. Acari numbers were similar between the Ingliston and Parwan locations (Table 1). Poduromorpha were most abundant at Parwan, accounting for two-thirds of the total invertebrates recorded from the site (Table 1). Notably, we found that Psocoptera were absent from Parwan, and no Araneae were recorded at Ingliston. There was a significant difference in the invertebrate composition among grass species and grass origin (Fig. 1 b and 1 c, respectively). Coleoptera & Diplopoda were present in exotic grasses in greater numbers. Conversely, Thysanoptera, Symphypleona and Entobryomorpha appeared in native grasses more frequently. Table 1. Mean abundance (± s.e.) of 12 invertebrate taxa, grouped by site and grass species. There was no significant difference in invertebrate composition between the small and large mesh bags (Fig. 1 a; Global R: 0.019; Pseudo p = 0.133). Figure 1 b illustrates the similarity of invertebrate composition based on the species of grass used in the mesh litterbags and shows that Themeda litter bags are more tightly grouped than other grass species, suggesting less variability in invertebrate composition. ANOSIM results provide evidence for a significant difference in invertebrate composition among the grass species (Global R = 0.134, Pseudo p = 0.001). In Fig. 1 c, the overlap of invertebrate composition between native and exotic grasses is clear. Nevertheless, the ANOSIM results provide evidence for a significant difference in invertebrate composition between the native and exotic grasses (Global R = 0.091, Pseudo p = 0.002). A clear grouping is evident among the three sites (Fig. 1 d), indicating that the site has the most substantial influence on the invertebrate composition (Global R = 0.491; pseudo p < 0.001). The Ingliston samples were more tightly clustered than the other two sites, suggesting greater similarity in invertebrate composition among the samples. SIMPER analysis (Table 2) revealed that Acari and Poduromorpha contributed the most to the dissimilarity between the Myrniong and Ingliston locations. Poduromorpha contributed the most dissimilarity between Myrniong and Parwan, and Ingliston and Parwan. Table 2 . Similarity percentage (SIMPER) partitions comparing taxa dissimilarity among sites. As the site was the most significant driver of invertebrate assemblages, the differences between the sites were further explored with abundance and ordinal diversity indices (Fig. 2 ). The mean invertebrate abundance at Parwan was significantly greater than the other two locations (Fig. 2 a), driven by several high-abundance litter bags. In contrast, Myrniong had significantly greater species diversity, evenness, and Margalef richness than the other two sites (Fig. 2 , p < 0.001 for each). Discussion Our study showed that in temperate grassland there is a diverse suite of invertebrate orders that will readily associate with detached grass litter. We found that the invertebrate community was most strongly influenced by the study location, suggesting there is an important level of spatial variation among litter-dwelling communities that may be driven by climate and abiotic differences among sites. Invertebrate community composition was also affected to a lesser extent by the species of grass and its origin but was not affected by the gauge of mesh bags used to collect samples. Grassland location We found that the sampling location had the strongest effect on invertebrate community composition. The literature suggests that vegetation structure (New 2000; Schultz et al. 2017), landscape context (Barton et al. 2024; Oliver et al. 2006), management history (Abraham & Morgan 2018; Bromham et al. 1999) and climate (Barnett & Facey 2016; Garcia-Palacios et al. 2013) may all exude a degree of influence on shaping invertebrate habitat. Several studies have suggested the sward structure or microhabitat influences the suitability for grassland fauna (Barton et al. 2024; Lindsay & Cunningham 2009; Price et al. 2019). Our three locations varied considerably in sward structure and available niches. Myrniong had a closed structure with dense Themeda growth, providing a sheltered aspect on a slight slope with good drainage. Ingliston had a short semi-open structure of grass and forbs, which was seasonally waterlogged in moist depressions. Parwan had an open sward structure, providing thermophilic habitat niches. Small forbs and rocks amongst tussocks added a layer of habitat complexity. These differences may explain the variation in invertebrate communities recorded. It would be useful to test for differences in litter-dwelling invertebrates among a larger range of sites that span a broad range of site variables to directly test these impacts. Grass origin The influence of native and exotic grass species on litter invertebrate community composition was significant in this study. This result was driven by the preferences of a select few orders: Coleoptera & Diplopoda were more frequently captured in exotic grass. Thysanoptera, Symphypleona and Entobryomorpha appeared in native grasses in higher numbers. As only five of the twelve orders demonstrated a preference towards grass origin, this result suggests generalists and specialists in the invertebrate fauna. Similar invertebrate responses in alien and native grass species have been noted elsewhere, albeit within the context of alien plant invasions (Belnap & Phillips 2001; Litt & Steidl 2010). Local observations note that exotic grass invasions can lead to a homogenised structure, and decreased habitat diversity (New 2019), whereas overall invertebrate abundance had a positive association with native grass cover in Australia’s grassy woodlands (Lindsay & Cunningham 2009). Grass species We found a significant difference in the invertebrates captured from the four grass species. Invertebrate responses to single grass species are poorly understood, at least locally. One Australian grassland study noted that some invertebrate species vacuum-sampled from live Themeda triandra and Poa labillardierei tussocks exhibited strong grass species preferences (Reid & Hochuli 2007). Plant traits such as nutrient status that influence litter variables can influence decomposition rate (Bradford et al. 2016; Cornwell et al. 2008). Australia’s native grasses have evolved for millennia in isolation (Bryceson & Morgan 2022) with dry climate, nutrient-poor soils, and frequent fire leading to a distinct assemblage of native flora (Orians & Milewski 2007). High lignin and low nitrogen content result in a recalcitrant litter that does not appear to follow the same decomposition trajectory as its northern hemisphere counterparts (Butler et al. 2023). Themeda and Poa differ in chemical composition, with Butler et al. (2023) demonstrating that Themeda decomposed at a greater rate than Poa over a 39-week period (Butler et al. 2023). Functional traits and nutrient status may have affected their suitability for invertebrate decomposers. Thysanoptera and Collembola (Entobryomorpha, Symphypleona and Poduromorpha) are two groups that have exhibited specialist plant associations, and this may have influenced our results. The endemic grass-dwelling members of Thysanoptera have been noted to have an association with Australian native grasses, including Themeda and Poa species, whereas exotic Thysanoptera taxa were associated with exotic grasses (Mound 2011). In Australia, Collembola has a high rate of endemism (Greenslade 2007), with exotic Collembola used as an indicator of disturbance. Exotic Collembola are largely absent from intact, low-nutrient, environments, suggesting disturbance and landscape change facilitate invasion (Greenslade 2018). Specialist fauna that has co-adapted to Australian flora may have impacted the invertebrates’ preferences for grass species within the study. The photosynthetic pathways of the grasses may also have influenced the invertebrate communities observed. Our study species were all C3 grasses except for Themeda , which is a C4 grass. We observed that Themeda showed lower variability in the invertebrate community among samples than the other grasses. This may be because C4 plants are generally thought to provide poorer food quality to decomposers due to low nutrient and high fibre content (Barbehenn & Bernays 1992; Scheirs et al. 2001; Scheunemann et al. 2010). Mesh size The use of two different mesh sizes in litter bags did not influence the invertebrate community sampled. Comparing our use of litterbags with other studies is challenging as previous research has focused on the disappearance rate of plant material rather than invertebrates associated with this process. While some studies have suggested that using a smaller mesh size excludes larger invertebrates (Delgado-Baquerizo et al. 2020; Handa et al. 2014; Kitz et al. 2015; Makkonen et al. 2012; Milcu & Manning 2011) and leads to slower decomposition rates, results from other studies (Cates et al. 2021; Schädler & Brandl 2005; Wall et al. 2008) make it difficult to draw definitive conclusions. Kampichler and Bruckner (2009) noted that comparing studies is challenging due to variations in mesh aperture, bag fillings, and the specific aims of the research. Our results suggest that the 2-mm mesh (which was easier to work with than the larger mesh size) was sufficient for observing the invertebrate community associated with decomposing grass litter that were the focus of this study. Conclusions Our study shows that the invertebrate community associated with grass litter is influenced by grass identity and grass origin. However, this influence is slight when compared to the differences emerging from separate locations. The differential response to the addition of exotic grass suggests that specialist invertebrates are present in the grass litter communities, and the encroachment of weeds may threaten the local persistence of invertebrates such as members of Entobryomorpha, Symphypleona and Thysanoptera orders, which appear to favour native grasses. The community variation among locations, even at a basic taxonomic level, emphasises the need for greater recognition of the role that invertebrates play in grassland ecosystems. Strategies aiming to improve grassland condition, such as maintaining structural diversity and managing grazing levels, generally benefit grassland invertebrates. However, our study shows significant variability within sites, making it challenging to assess these benefits at specific locations. Therefore, improving monitoring of grassland invertebrates will help us measure the outcomes of management practices. In turn, this will allow us to anticipate how grasslands might respond to global change and develop strategies for conserving invertebrate diversity. Declarations Conflict of Interest. The authors declare no conflict of interest. Author Contribution All authors contributed to the study conception and design, material preparation, data collection and analysis. AMW wrote the original draft manuscript and prepared table 2. NLS prepared figures 1-2. All authors reviewed and edited the manuscript and approved the final manuscript. Acknowledgement Allison would like to acknowledge the memory of Roslynn Shearer, whose encouragement, friendship, and support contributed to the fruition of this research. Thanks to Tom Mills for assistance with fieldwork, and Penelope Greenslade for input into initial discussions. We are grateful to Simon & Lorraine Jolly for allowing the use of their property. Thanks to Tom Miller and the Moorabool Shire for allowing the use of further study locations. Finally, we acknowledge and pay our respects to the Wadawurrung and Wurundjeri Traditional Owners of the land on which this research took place. Data Availability Data is provided within the supplementary information files. References Abraham J, Morgan JW (2018) Effects of time-since-fire on the invertebrate communities of Kangaroo grass “Themeda triandra” -dominated grasslands in Melbourne, Victoria. The Vic Naturalist 135:36-46. https://search.informit.org/doi/10.3316/informit.350603692010878 Aerts R (1997) Climate, leaf litter chemistry and leaf litter decomposition in terrestrial ecosystems: a triangular relationship. Oikos 79:439-449. https://doi.org/10.2307/3546886 Antos M, Williams N (2015) The wildlife of our grassy landscape. In: N Williams, A Marshall & J Morgan (eds), Land of Sweeping Plains: Managing and Restoring the Native Grasslands of south-eastern Australia. CSIRO Publishing, Melbourne, pp 87-114 Austin AT, Vivanco L (2006) Plant litter decomposition in a semi-arid ecosystem controlled by photodegradation. Nat 442:555-558. https://doi.org/10.1038/nature05038 Bagyaraj DJ, Nethravathi CJ, Nitin KS (2016) Soil Biodiversity and Arthropods: Role in Soil Fertility. In: AK Chakravarthy & S Sridhara (eds), Economic and Ecological Significance of Arthropods in Diversified Ecosystems: Sustaining Regulatory Mechanisms. Springer Singapore, Singapore, pp 17-51 Barbehenn RV, Bernays EA (1992) Relative nutritional quality of C 3 and C 4 grasses for a graminivorous lepidopteran, Paratrytone melane (Hesperiidae). Oecologia 92:97-103. https://doi.org/10.1007/BF00317268 Barnett KL, Facey SL (2016) Grasslands, Invertebrates, and Precipitation: A Review of the Effects of Climate Change. Front Plant Sci 7:1196. https://doi.org/10.3389/fpls.2016.01196 Barton PS, Evans MJ (2017) Insect biodiversity meets ecosystem function: differential effects of habitat and insects on carrion decomposition. Ecol Entomol 42:364-74. https://doi.org/10.1111/een.12395 Barton PS, Evans MJ, Lewis J (2024) Microhabitats shape ant community structure in a spatially heterogeneous grassy woodland. Ecosphere 15:4798. https://doi.org/10.1002/ecs2.4798 Belnap J, Phillips SL (2001) Soil biota in an ungrazed grassland: response to annual grass ( Bromus tectorum ) invasion. Ecol Appl 11:1261-75. https://doi.org/10.1890/1051-0761(2001)011[1261:SBIAUG]2.0.CO;2 Bradford MA, Berg B, Maynard DS, Wieder WR, Wood, SA (2016) Future Directions: Understanding the dominant controls on litter decomposition. J Ecol 104:229-38. http://www.jstor.org/stable/24762886 Bromham L, Cardillo M, Bennett AF, Elgar, MA (1999) Effects of stock grazing on the ground invertebrate fauna of woodland remnants. Aust J Ecol 24:199-207. https://doi.org/10.1046/j.1442-9993.1999.00963.x Bryceson SR, Morgan JW (2022) The Australasian grass flora in a global context. J Syst Evol 60:675-690. https://doi.org/10.1111/jse.12839 Butler F, Good M, Morgan J, Schultz N (2023) Relative contribution of photodegradation to litter breakdown in Australian grasslands. Ecol Evol 13:10710. https://doi.org/10.1002/ece3.10710 Cardoso P, Erwin TL, Borges PA, New TR (2011) The seven impediments in invertebrate conservation and how to overcome them. Biol Conserv 144:2647-2655. https://doi.org/10.1016/j.biocon.2011.07.024 Cates AM, Wills BD, Kim TN, Landis DA, Gratton C, Read HW, Jackson RD (2021) No evidence of top-down effects by ants on litter decomposition in a temperate grassland. Ecosphere 12:03638. https://doi.org/10.1002/ecs2.3638 Chapman AD (2009) Numbers of living species in Australia and the World, 2nd edn. Department of the Environment, Water, Heritage and the Arts, Australian Government, Canberra. Clarke K, Gorley R (2013) Primer 6: Version 6.1.16. User Manual/Tutorial. Primer-E Ltd, Plymouth Clarke KR (1993) Non‐parametric multivariate analyses of changes in community structure. Aust J Ecol 181:117-43. https://doi.org/10.1111/j.1442-9993.1993.tb00438.x Clay RE (2014) Potential Effects of the Loss of Native Grasses on Grassland Invertebrate Diversity in Southeastern Australia. Int J Ecol 2014:202056. https://doi.org/10.1155/2014/202056 Coleman DC, Callaham MA, Crossley DA (2017) Fundamentals of Soil Ecology, 3rd edn. Academic Press, London. Cornwell WK, Cornelissen JH, Amatangelo K, Dorrepaal E, Eviner VT, Godoy O, Hobbie SE, Hoorens B, Kurokawa H, Perez-Harguindeguy N (2008) Plant species traits are the predominant control on litter decomposition rates within biomes worldwide, Ecol Lett 11:1065-1071. https://doi.org/10.1111/j.1461-0248.2008.01219.x Cuevas E, Medina E (1988) Nutrient dynamics within amazonian forests: II. Fine root growth, nutrient availability and leaf litter decomposition. Oecologia 76:222-235. https://doi.org/10.1007/BF00379956 Culliney TW (2013) Role of Arthropods in Maintaining Soil Fertility. Agriculture 3:629-659 https://doi.org/10.3390/agriculture3040629 Curry J (1987) The invertebrate fauna of grassland and its influence on productivity. III. Effects on soil fertility and plant growth. Grass Forage Sci 42:325-341 https://doi.org/10.1111/j.1365-2494.1987.tb02121.x Daley A, Ellingsen K (2012) Insects of Tasmania: An online field guide. https://tasmanianinsectfieldguide.com/. Accessed 25 April 2023 Decaëns T (2010) Macroecological patterns in soil communities. Global Ecol Biogeogr 19:287-302. https://doi.org/10.1111/j.1466-8238.2009.00517.x Delgado-Baquerizo M, Reich PB, Trivedi C, Eldridge DJ, Abades S, Alfaro FD, Bastida F, Berhe AA, Cutler NA, Gallardo A, García-Velázquez L, Hart SC, Hayes PE, He J-Z, Hseu Z-Y, Hu H-W, Kirchmair M, Neuhauser S, Pérez CA, Reed SC, Santos F, Sullivan BW, Trivedi P, Wang J-T, Weber-Grullon L, Williams MA, Singh BK (2020) Multiple elements of soil biodiversity drive ecosystem functions across biomes. Nat Ecol Evol 4:210-20. https://doi.org/10.1038/s41559-019-1084-y Fay PA, Prober SM, Harpole WS, Knops JMH, Bakker JD, Borer ET, Lind EM, MacDougall AS, Seabloom EW, Wragg PD, Adler PB, Blumenthal DM, Buckley YM, Chu C, Cleland EE, Collins SL, Davies KF, Du G, Feng X, Firn J, Gruner DS, Hagenah N, Hautier Y, Heckman RW, Jin VL, Kirkman KP, Klein J, Ladwig LM, Li Q, McCulley RL, Melbourne BA, Mitchell CE, Moore JL, Morgan JW, Risch AC, Schütz M, Stevens CJ, Wedin DA, Yang LH (2015) Grassland productivity limited by multiple nutrients. Nat Plants 1:15080. http://dx.doi.org/10.1038/nplants.2015.80 Garcia-Palacios P, Maestre FT, Kattge J, Wall DH (2013) Climate and litter quality differently modulate the effects of soil fauna on litter decomposition across biomes. Ecol Lett 16:1045-1053. https://doi.org/10.1111/ele.12137 Gessner MO, Swan CM, Dang CK, McKie BG, Bardgett RD, Wall DH, Hättenschwiler S (2010) Diversity meets decomposition. Trends Ecol Evol 25:372-880. https://doi.org/10.1016/j.tree.2010.01.010 Gibson L, New T (2007) Characterising insect diversity on Australia’s remnant native grasslands: ants (Hymenoptera: Formicidae) and beetles (Coleoptera) at Craigieburn Grasslands Reserve, Victoria. J Insect Conserv 11:409-413. https://doi.org/10.1007/s10841-006-9051-8 Gongalsky KB (2021) Soil macrofauna: Study problems and perspectives. Soil Biol Biochem 159:108281. https://doi.org/10.1016/j.soilbio.2021.108281 Gooderham J, Tsyrlin E (2002) The Waterbug Book: A Guide to the Freshwater Macroinvertebrates of Temperate Australia. CSIRO Publishing, Melbourne. Greenslade P (1997) Short term effects of a prescribed burn on invertebrates in grassy woodland in southeastern Australia. Memoirs of Museum Victoria 56:305-312. https://doi.org/10.24199/j.mmv.1997.56.18 Greenslade P (2007) The potential of Collembola to act as indicators of landscape stress in Australia. Aust J Exp Agr 47: 424-34. https://doi.org/10.1071/EA05264 Greenslade P (2018) Why are there so many exotic Springtails in Australia? A review. Soil organisms 90:141-156. https://doi.org/10.25674/y9tz-1d49 Griffiths HM, Ashton LA, Parr CL, Eggleton P (2021) The impact of invertebrate decomposers on plants and soil. New Phytol 231:2142-2149. https://doi.org/10.1111/nph.17553 Handa IT, Aerts R, Berendse F, Berg MP, Bruder A, Butenschoen O, Chauvet E, Gessner MO, Jabiol J, Makkonen, M (2014) Consequences of biodiversity loss for litter decomposition across biomes. Nat 509:218-221. https://doi.org/10.1038/nature13247 Hättenschwiler S, Tiunov AV, Scheu S (2005) Biodiversity and Litter Decomposition in Terrestrial Ecosystems. Annu Rev Ecol Evol S 36:191-218. https://doi.org/10.1146/annurev.ecolsys.36.112904.151932 Heděnec P, Jiménez JJ, Moradi J, Domene X, Hackenberger D, Barot S, Frossard A, Oktaba L, Filser, J, Kindlmann P (2022) Global distribution of soil fauna functional groups and their estimated litter consumption across biomes. Sci Rep 12:17362. https://doi.org/10.1038/s41598-022-21563-z Hungate BA, Barbier EB, Ando AW, Marks SP, Reich PB, Van Gestel N, Tilman D, Knops JM, Hooper DU, Butterfield, BJ (2017) The economic value of grassland species for carbon storage. Sci Advances 3:1601880 DOI: 10.1126/sciadv.1601880 Kampichler C, Bruckner A (2009) The role of microarthropods in terrestrial decomposition: a meta‐analysis of 40 years of litterbag studies. Biol Rev 84:375-389. https://doi.org/10.1111/j.1469-185X.2009.00078.x Kassambara A (2023) ggpubr: 'ggplot2' Based Publication Ready Plots, R package version 0.6.0, https://rpkgs.datanovia.com/ggpubr/ Accessed 8 May 2023 King KL, Hutchinson KJ (1983) The effects of sheep grazing on invertebrate numbers and biomass in unfertilized natural pastures of the New England Tablelands (NSW). Aust J Ecol 8: 245-255. https://doi.org/10.1111/j.1442-9993.1983.tb01322.x Kitz F, Steinwandter M, Traugott M, Seeber J (2015) Increased decomposer diversity accelerates and potentially stabilises litter decomposition. Soil Biol Biochem 83:138-141. https://doi.org/10.1016/j.soilbio.2015.01.026 Knops JM, Wedin D, Tilman D (2001) Biodiversity and decomposition in experimental grassland ecosystems. Oecologia 126:429-433. https://doi.org/10.1007/s004420000537 Lavelle P, Decaëns T, Aubert M, Barot Sb, Blouin M, Bureau F, Margerie P, Mora P, Rossi, J-P (2006) Soil invertebrates and ecosystem services. Eur J Soil Biol 42:S3-S15. https://doi.org/10.1016/j.ejsobi.2006.10.002 Lindsay EA, Cunningham SA (2009) Livestock grazing exclusion and microhabitat variation affect invertebrates and litter decomposition rates in woodland remnants. Forest Ecol Manag 258:178-187. https://doi.org/10.1016/j.foreco.2009.04.005 Litt AR, Steidl RJ (2010) Insect assemblages change along a gradient of invasion by a nonnative grass. Biol Invasions 12:3449-3463. https://doi.org/10.1007/s10530-010-9743-6 Makkonen M, Berg MP, Handa IT, Hättenschwiler S, van Ruijven J, van Bodegom PM, Aerts R (2012) Highly consistent effects of plant litter identity and functional traits on decomposition across a latitudinal gradient. Ecol Lett 15:1033-1041. https://doi.org/10.1111/j.1461-0248.2012.01826.x Marshall D, Fitzsimons JA (2008) Challenges for native grassland conservation on Victoria's Northern Plains. Australas Plant Conserv 16:24-5. https://search.informit.org/doi/10.3316/informit.023279248905647 Milcu A, Manning P (2011) All size classes of soil fauna and litter quality control the acceleration of litter decay in its home environment. Oikos 120:1366-1370. https://doi.org/10.1111/j.1600-0706.2010.19418.x Miller L, New T (1997) Mount Piper grasslands: pitfall trapping of ants and interpretation of habitat variability. Memoirs of the Museum of Victoria 56:377-381 Morgan JW (1998) Patterns of invasion of an urban remnant of a species-rich grassland in southeastern Australia by non-native plant species. J Veg Sci 9:181-190. https://doi.org/10.2307/3237117 Morris MG (2000) The effects of structure and its dynamics on the ecology and conservation of arthropods in British grasslands. Biol Conserv 95:129-142. https://doi.org/10.1016/S0006-3207(00)00028-8 Mound LA (2011) Grass-dependent Thysanoptera of the family Thripidae from Australia, Zootaxa 3064:1-40. https://doi.org/10.11646/zootaxa.3064.1.1 New TR (2000) How Useful are Ant Assemblages for Monitoring Habitat Disturbance on Grasslands in South Eastern Australia? J Insect Conserv 4:153-159. https://doi.org/10.1023/A:1009668817271 New TR (2019) Insect Conservation and Australia’s Grasslands. Springer Nature, Cham, Switzerland. https://doi.org/10.1007/978-3-030-22780-7 Oksanen J, Blanchet FG, Friendly M, Kindt R, Legendre P, McGlinn, D, Minchin PR, O’Hara RB, Simpson GL, Solymos P, Stevens MHH, Szoecs E, Wagner H (2019) Vegan: Community Ecology Package. R Package Version 2.5–6. https://CRAN.R-project.org/package=vegan. Oliver I, Pearce S, Greenslade PJ & Britton, DR (2006) Contribution of paddock trees to the conservation of terrestrial invertebrate biodiversity within grazed native pastures. Austral Ecol 31:1-12. https://doi.org/10.1111/j.1442-9993.2006.01537.x Orgiazzi A, Bardgett R, Barrios E, Behan-Pelletier V, Briones M, Chotte J, De Deyn G, Eggleton P, Fierer N, Fraser T, Hedlund K, Jeffery S, Johnson N, Jones A, Kandeler E, Kaneko N, Lavelle P, Lemanceau P, Miko L, Montanarella L, Moreira F, Ramirez K, Scheu S, Singh B, Six J, van der Putten W, Wall DE (2016) Global Soil Biodiversity Atlas, Publications Office of the European Union, Luxembourg. Orians GH, Milewski AV (2007) Ecology of Australia: the effects of nutrient-poor soils and intense fires. Biol Rev 82:393-423. https://doi.org/10.1111/j.1469-185X.2007.00017.x Patoine G, Thakur MP, Friese J, Nock C, Hönig L, Haase J, Scherer-Lorenzen M, Eisenhauer N (2017) Plant litter functional diversity effects on litter mass loss depend on the macro-detritivore community. Pedobiologia 65:29-42. https://doi.org/10.1016/j.pedobi.2017.07.003 Price JN, Good MK, Schultz NL, Guja LK, Morgan JW (2019) Multivariate drivers of diversity in temperate Australian native grasslands. Aust J Bot 67:367-380. https://doi.org/10.1071/BT18190 R Core Team (2023) R: A Language and Environment for Statistical Computing, The R Foundation for Statistical Computing, Vienna, Austria. https://www.r-project.org/ Reid, AM & Hochuli, DF (2007) Grassland invertebrate assemblages in managed landscapes: Effect of host plant and microhabitat architecture. Austral Ecol 32:708-718. https://doi.org/10.1111/j.1442-9993.2007.01767.x Rosenberg Y, Bar-On YM, Fromm A, Ostikar M, Shoshany A, Giz O, Milo R (2023) The global biomass and number of terrestrial arthropods. Sci Advances 9:eabq4049 https://doi.org/10.1126/sciadv.abq4049 Ross CE, Barton PS, McIntyre S, Cunningham SA, Manning AD (2017) Fine‐scale drivers of beetle diversity are affected by vegetation context and agricultural history. Austral Ecol 42:831-843. https://doi.org/10.1111/aec.12506 Roy PG, Delpratt J (2006) Victorian Basalt Plains Grasslands - an Overview. Australas Plant Conserv 15:4-6. https://search.informit.org/doi/10.3316/informit.054675805475714 Samways MJ, Caldwell P, Osborn R (1996) Ground-living invertebrate assemblages in native, planted and invasive vegetation in South Africa. Agr Ecosyst Environ 59:19-32. https://doi.org/10.1016/0167-8809(96)01047-X Saunders ME, Barton PS, Bickerstaff JRM, Frost L, Latty T, Lessard BD, Lowe EC, Rodriguez J, White TE, Umbers KDL (2021) Limited understanding of bushfire impacts on Australian invertebrates. Insect Conserv Diver 14:285-293. https://doi.org/10.1111/icad.12493 Schädler M, Brandl R (2005) Do invertebrate decomposers affect the disappearance rate of litter mixtures? Soil Biol Biochem 37:329-337. https://doi.org/10.1016/j.soilbio.2004.07.042 Scheirs J, De Bruyn L, Verhagen R (2001) A test of the C3–C4 hypothesis with two grass miners. Ecol. 82:410-421 https://doi.org/10.1016/j.soilbio.2004.07.042 Scheunemann N, Scheu S, Butenschoen O (2010) Incorporation of decade old soil carbon into the soil animal food web of an arable system. Appl Soil Ecol 46:59-63. https://doi.org/10.1016/j.apsoil.2010.06.014 Schultz N, Keatley M, Antos M, Wong N, Moxham C, Farmilo B, Morgan JW (2017) The golf ball method for rapid assessment of grassland structure. Ecol Manag Restor 18:134-140. https://doi.org/10.1111/emr.12254 Solascasas P, Azcárate FM, Hevia V (2022) Edaphic arthropods as indicators of the ecological condition of temperate grassland ecosystems: A systematic review. Ecol Indic 142:109277. https://doi.org/10.1016/j.ecolind.2022.109277 Swift MJ, Heal OW, Anderson, JM (1979) Decomposition in terrestrial ecosystems, vol. 5, Blackwell Scientific, Oxford. Taylor GS, Braby MF, Moir ML, Harvey MS, Sands DP, New TR, Kitching RL, McQuillan PB, Hogendoorn K, Glatz RV, Andren M, Cook JM, Henry SC, Valenzuela I, Weinstein P (2018) Strategic national approach for improving the conservation management of insects and allied invertebrates in Australia. Austral Entomol 57:124-149. https://doi.org/10.1111/aen.12343 Wall DH, Bradford MA, St. John MG, Trofymow JA, Behan‐Pelletier V, Bignell DE, Dangerfield JM, Parton WJ, Rusek J, Voigt W (2008) Global decomposition experiment shows soil animal impacts on decomposition are climate‐dependent. Global Change Biol 14:2661-2677. https://doi.org/10.1111/j.1365-2486.2008.01672.x White RP, Murray S, Rohweder M, Prince S, Thompson K (2000) Grassland Ecosystems, World Resources Institute, Washington, DC, USA. Yang Y, Tilman D, Furey G, Lehman C (2019) Soil carbon sequestration accelerated by restoration of grassland biodiversity. Nat Commun 10:718. https://doi.org/10.1038/s41467-019-08636-w Yen A (1999) Grassland invertebrates of the western Victorian basalt plains: plant crunchers or forgotten lunches. In ‘The Great Plains crash: Proceedings of a conference on the grasslands and grassy woodlands of Victoria’. Victorian Institute of Technology (Ed. RN Jones) pp. 57-68. Indigenous Flora and Fauna Association/ Victorian National Parks Association, East Melbourne, Victoria Yoshioka A, Kadoya T, Suda S-I, Washitani I (2010) Impacts of weeping lovegrass ( Eragrostis curvula ) invasion on native grasshoppers: responses of habitat generalist and specialist species. Biol Invasions 12:531-539. https://doi.org/10.1007/s10530-009-9456-x Zborowski P, Storey R (2017) A Field Guide to Insects in Australia, 4 edn. New Holland, Sydney. Zeng X, Gao H, Wang R, Majcher BM, Woon JS, Wenda C, Eggleton P, Griffiths HM, Ashton LA (2024) Global contribution of invertebrates to forest litter decomposition. Ecol Lett 27:e14423. https://doi.org/10.1111/ele.14423 Tables Table 1 and 2 are available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files SuppinfoS1.docx Suppinfo2GrasslandinvertrawdataWall.csv Table1AND2.docx Cite Share Download PDF Status: Published Journal Publication published 07 Apr, 2025 Read the published version in Journal of Insect Conservation → Version 1 posted Editorial decision: Revision requested 06 Jan, 2025 Reviews received at journal 23 Dec, 2024 Reviews received at journal 19 Dec, 2024 Reviews received at journal 17 Dec, 2024 Reviewers agreed at journal 17 Dec, 2024 Reviewers agreed at journal 16 Dec, 2024 Reviewers agreed at journal 16 Dec, 2024 Reviewers agreed at journal 16 Dec, 2024 Reviews received at journal 15 Dec, 2024 Reviewers agreed at journal 10 Dec, 2024 Reviewers invited by journal 10 Dec, 2024 Editor assigned by journal 20 Nov, 2024 Submission checks completed at journal 20 Nov, 2024 First submitted to journal 19 Nov, 2024 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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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-5486939\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":380532670,\"identity\":\"04e4f8c9-f366-4d31-b6a6-dfcec5ae358e\",\"order_by\":0,\"name\":\"Allison M. Wall\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Federation University Australia\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Allison\",\"middleName\":\"M.\",\"lastName\":\"Wall\",\"suffix\":\"\"},{\"id\":380532671,\"identity\":\"4b88bd1d-f01e-4eca-a14a-f61510cdc7a3\",\"order_by\":1,\"name\":\"Philip S. 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Schultz\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABEklEQVRIie3QwUrDMBjA8a8EmstXev1Kkb5Cx2AqU/YqKQV7EVQE2WFIvLSXwq4O9xA+ghCol+IjSIew0w6DHQfD0A1Pqe7oIX9IIIQfIR+AzfZPYwBnAPypATwcf/ZfCBGgiluCxxNKDwT+IKfAF5u7nB6j2ZfTrMaf0YizqoHxMJG8jk3kXGI/nOVEzkvKevP6vlcyN4uhzhKJ10YSv6HLPE1YeFOFXi6ckuGAnFwlEroIX7bEDRTfejsx2pOdJv6qg8CgJUhMPydFsidSE+p4Rem/4AcFz5iyYF6JtFTuFYkq6+e0vDWS92KxwYcLPyqUs15NxGUxVRWtJ8OTqZ++Gqdsnr/QyzXe2Gw2m+2YvgEPiU6ynIlLKAAAAABJRU5ErkJggg==\",\"orcid\":\"\",\"institution\":\"Federation University Australia\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Nick\",\"middleName\":\"L.\",\"lastName\":\"Schultz\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2024-11-20 02:23:24\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-5486939/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-5486939/v1\",\"draftVersion\":[],\"editorialEvents\":[{\"content\":\"https://doi.org/10.1007/s10841-025-00668-6\",\"type\":\"published\",\"date\":\"2025-04-07T16:04:58+00:00\"}],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":70451502,\"identity\":\"77da783e-95d6-4fb1-8f8e-96200efb348f\",\"added_by\":\"auto\",\"created_at\":\"2024-12-03 09:48:05\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":235460,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eNon-metric multidimensional scaling ordination diagrams showing the compositional difference among invertebrate communities grouped by (a) mesh aperture, (b) grass species, (c) native versus exotic grass species origin, and (d) the three grassland sites. The global R and pseudo p values returned by ANOSIM are provided for each panel, reflecting the significance of group differences.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5486939/v1/0a7b4aeb8746759a6fb37c74.png\"},{\"id\":70452658,\"identity\":\"40abb899-22d1-4551-a61a-0a0f686b5989\",\"added_by\":\"auto\",\"created_at\":\"2024-12-03 09:56:05\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":66587,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eBoxplot graphs showing the mean, interquartile range, and minimum/maximum values of invertebrate abundance at each site (A), and three measures of ordinal diversity (B, C, D) across the study sites. Dots represent individual values from the 24 samples at each site.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"OnlineFigure2divmeasures.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5486939/v1/eae883b3ec0a34020e07f131.png\"},{\"id\":80559076,\"identity\":\"3f715ca0-2ce7-4d08-9909-003c6e57b819\",\"added_by\":\"auto\",\"created_at\":\"2025-04-14 16:17:41\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":801869,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5486939/v1/5c13a555-0370-443c-ab3f-f28728e2e0a5.pdf\"},{\"id\":70451506,\"identity\":\"2423d002-362b-493c-9dcf-a2350bb3baea\",\"added_by\":\"auto\",\"created_at\":\"2024-12-03 09:48:06\",\"extension\":\"docx\",\"order_by\":1,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":3242843,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"SuppinfoS1.docx\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5486939/v1/42f6d8579d94ce612b0c83bd.docx\"},{\"id\":70451505,\"identity\":\"455eba98-d2ef-4b36-9a29-6e7bcb801cfe\",\"added_by\":\"auto\",\"created_at\":\"2024-12-03 09:48:05\",\"extension\":\"csv\",\"order_by\":2,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":43893,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"Suppinfo2GrasslandinvertrawdataWall.csv\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5486939/v1/efc99deecdf5253a8d65ccdb.csv\"},{\"id\":70453024,\"identity\":\"fd4e9e8c-0da9-49e1-bafb-0de5f70ac9e1\",\"added_by\":\"auto\",\"created_at\":\"2024-12-03 10:04:05\",\"extension\":\"docx\",\"order_by\":3,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":421443,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"Table1AND2.docx\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-5486939/v1/ea54bf6157d70558be95fb15.docx\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"Litter invertebrates display greater differences among locations than grass species in a temperate grassland\",\"fulltext\":[{\"header\":\"Introduction\",\"content\":\"\\u003cp\\u003eDecomposition in grasslands is a crucial process contributing to soil formation and global carbon cycles (Hungate et al. \\u003cspan citationid=\\\"CR42\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e; Yang et al. \\u003cspan citationid=\\\"CR82\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e), with an estimated 34% of terrestrial carbon stored in grassland ecosystems (White et al. \\u003cspan citationid=\\\"CR81\\\" class=\\\"CitationRef\\\"\\u003e2000\\u003c/span\\u003e). Up to 90% of terrestrial plant matter is decomposed rather than consumed in some ecosystems (Gessner et al. \\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e). The factors that drive plant decomposition include climate variability (Wall et al. \\u003cspan citationid=\\\"CR80\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e), photodegradation (Austin \\u0026amp; Vivanco \\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e2006\\u003c/span\\u003e), nutrient availability (Bradford et al. \\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e; Cuevas \\u0026amp; Medina \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e1988\\u003c/span\\u003e), and the interacting effects of these factors (Aerts \\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e1997\\u003c/span\\u003e; Garcia-Palacios et al. \\u003cspan citationid=\\\"CR30\\\" class=\\\"CitationRef\\\"\\u003e2013\\u003c/span\\u003e; Makkonen et al. \\u003cspan citationid=\\\"CR51\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e). Invertebrates also play a key role in accelerating plant litter decomposition (Heděnec et al. \\u003cspan citationid=\\\"CR41\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e; Wall et al. \\u003cspan citationid=\\\"CR80\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e), but we know little about the local factors influencing invertebrate community composition in many grasslands (Gibson \\u0026amp; New \\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e2007\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eInvertebrates are an often-overlooked component of fauna due to their overwhelming diversity and lack of expertise among conservation practitioners (Cardoso et al. \\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e2011\\u003c/span\\u003e). Invertebrate decomposers are a particularly important group due to their contribution to ecosystem function by facilitating biomass comminution, conversion and nutrient exchange (Handa et al. \\u003cspan citationid=\\\"CR39\\\" class=\\\"CitationRef\\\"\\u003e2014\\u003c/span\\u003e; H\\u0026auml;ttenschwiler et al. \\u003cspan citationid=\\\"CR40\\\" class=\\\"CitationRef\\\"\\u003e2005\\u003c/span\\u003e; Swift et al.1979). Invertebrates play an essential role in breaking down plant litter, dung, and carrion and returning nutrients to the soil and wider biosphere (Barton \\u0026amp; Evans \\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e; Bradford et al. \\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e; Griffiths et al. \\u003cspan citationid=\\\"CR38\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; Lavelle et al. \\u003cspan citationid=\\\"CR48\\\" class=\\\"CitationRef\\\"\\u003e2006\\u003c/span\\u003e). Temperate grasslands are disproportionately rich in decomposer invertebrates and can support up to 100 tonnes per hectare of earthworms, microarthropods, fungi and bacteria (Orgiazzi et al. \\u003cspan citationid=\\\"CR62\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e), driving high rates of litter consumption (Heděnec et al. \\u003cspan citationid=\\\"CR41\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eThe stratum of leaf litter covering the topsoil is the initial habitat in which macro arthropods break down litter (Coleman et al. \\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e). Tearing, shredding, ingesting and excreting mass results in weakened litter and smaller particles and a more labile state (Gongalsky \\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; Patoine et al. \\u003cspan citationid=\\\"CR64\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e). Acari and Collembola are considered the main litter transformers responsible for the break down and mixing of plant litter (Culliney \\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e2013\\u003c/span\\u003e), with Millipedes, Isopods, and Insects also contributing. Detritivores produce enzymes to break down plant material, weakening and fragmenting plant litter with ingestion (Griffiths et al. \\u003cspan citationid=\\\"CR38\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e). These can include mites, millipedes and the larvae of Diptera and Coleoptera (Curry \\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e1987\\u003c/span\\u003e), as well as up to 20,000 Collembola per square metre (Greenslade \\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e2007\\u003c/span\\u003e). The movement of these arthropods as they forage and consume dead organic matter results in the subsoil becoming aerated and soil and nutrient mixing (Bagyaraj et al. \\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e). Microarthropods most frequently inhabit the top 5cm of grassland ecosystems (Coleman et al. \\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e). A loss of diversity in invertebrates and grass species has been shown to slow the cycling of carbon and nitrogen from leaf litter across several biomes (Handa et al. \\u003cspan citationid=\\\"CR39\\\" class=\\\"CitationRef\\\"\\u003e2014\\u003c/span\\u003e) and in controlled laboratory experiments (Delgado-Baquerizo et al. \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). The sensitivity of litter-dwelling arthropods has suggested they are a valuable measure of ecosystem health in temperate grasslands (Solascasas et al. \\u003cspan citationid=\\\"CR77\\\" class=\\\"CitationRef\\\"\\u003e2022\\u003c/span\\u003e), with population fluxes rapidly responding to human management practices (Greenslade \\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e2007\\u003c/span\\u003e). Despite the importance of arthropods in the initial breakdown of leaf litter, this is a rarely studied topic, and specific data on local scales is absent from many ecosystems (H\\u0026auml;ttenschwiler et al. \\u003cspan citationid=\\\"CR40\\\" class=\\\"CitationRef\\\"\\u003e2005\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eComparing invertebrate assemblages in native and exotic grass species can predict how ecosystems will respond to the invasion of alien grass species or be affected by degradation. Belnap \\u0026amp; Phillips (\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e2001\\u003c/span\\u003e) showed how the invasion of an exotic grass resulted in lower species richness and overall invertebrate abundance. A similar decline in local insect diversity was recorded by Litt and Steidl (\\u003cspan citationid=\\\"CR50\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e) in \\u003cem\\u003eEragrostis lehmanniana\\u003c/em\\u003e-invaded native pastures in Arizona, though a few orders favoured the change. Similar trends in response to invasive species have been noted in South Africa (Samways et al. \\u003cspan citationid=\\\"CR71\\\" class=\\\"CitationRef\\\"\\u003e1996\\u003c/span\\u003e), and South Australia (Clay \\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e2014\\u003c/span\\u003e) as well as the increased susceptibility of specialist invertebrate species compared to generalists (Yoshioka et al. \\u003cspan citationid=\\\"CR84\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e). In Australian temperate grasslands, Miller and New (\\u003cspan citationid=\\\"CR54\\\" class=\\\"CitationRef\\\"\\u003e1997\\u003c/span\\u003e) explored the change in ant assemblages in intact and degraded sites and found that while the composition of the ant community varied, the composition of functional groups did not (Miller \\u0026amp; New \\u003cspan citationid=\\\"CR54\\\" class=\\\"CitationRef\\\"\\u003e1997\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eLittle research has been published on grassland invertebrates in southern Australia, and no systematic study has been undertaken on Victoria\\u0026rsquo;s grasslands (Yen \\u003cspan citationid=\\\"CR83\\\" class=\\\"CitationRef\\\"\\u003e1999\\u003c/span\\u003e). This knowledge gap concerns ecologists as 70% of insect endemism is estimated in Australia (Chapman \\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e2009\\u003c/span\\u003e), and less than half of the insect fauna is described (Saunders et al. \\u003cspan citationid=\\\"CR72\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; Zborowski \\u0026amp; Storey \\u003cspan citationid=\\\"CR85\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e). Further evidence is needed on the role of invertebrates in decomposition and how the nutrient status of grass species mediates this (Delgado-Baquerizo et al. \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). We cannot effectively manage our current remnants without understanding the influence of invertebrates, and how transitions towards a more alien assemblage of plants might impact the efficacy of plant breakdown and associated nutrient cycling. Furthermore, knowledge gaps remain regarding the identity of the invertebrates that break down litter (Rosenberg et al. \\u003cspan citationid=\\\"CR68\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e). This shortfall has management repercussions, making us unable to predict the impact of exotic plant invasions, exotic invertebrates, and climate change (Deca\\u0026euml;ns \\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003eThe range of natural grasslands in southern Australia is heavily depleted, with \\u0026lt;\\u0026thinsp;5% of the once extensive ecosystem now considered intact (Roy \\u0026amp; Delpratt \\u003cspan citationid=\\\"CR70\\\" class=\\\"CitationRef\\\"\\u003e2006\\u003c/span\\u003e). Many grasslands in Australia now feature naturalised, self-sustaining populations of exotic grass (Morgan \\u003cspan citationid=\\\"CR55\\\" class=\\\"CitationRef\\\"\\u003e1998\\u003c/span\\u003e; New \\u003cspan citationid=\\\"CR59\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e), which has several implications for grasslands\\u0026rsquo; function and ecological processes, including decomposition. The native grasses of Australia are adapted to low-nutrient and shallow soils (Fay et al. \\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e; King \\u0026amp; Hutchinson \\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e1983\\u003c/span\\u003e; Orians \\u0026amp; Milewski \\u003cspan citationid=\\\"CR63\\\" class=\\\"CitationRef\\\"\\u003e2007\\u003c/span\\u003e). The grass decomposition trajectories may differ from those of higher-nutrient systems, with the photodegradation of high-lignin grasses acting as a primer for further decomposition (Butler et al. \\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e). Nevertheless, the influence of leaf litter types on the local invertebrate decomposer community has not been tested.\\u003c/p\\u003e\\n\\u003ch3\\u003eAims\\u003c/h3\\u003e\\n\\u003cp\\u003eWe aimed to determine which invertebrate orders are present in the initial breakdown of detached grass litter. Within this context, we asked:\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e\\u003col\\u003e \\u003cspan\\u003e \\u003cli\\u003e \\u003cp\\u003eDo invertebrate orders exhibit a preference for particular grass species?\\u003c/p\\u003e \\u003c/li\\u003e \\u003c/span\\u003e \\u003cspan\\u003e \\u003cli\\u003e \\u003cp\\u003eDo invertebrates respond differently to native and exotic grass species?\\u003c/p\\u003e \\u003c/li\\u003e \\u003c/span\\u003e \\u003cspan\\u003e \\u003cli\\u003e \\u003cp\\u003eDoes the mesh aperture exclude elements of the invertebrates present?\\u003c/p\\u003e \\u003c/li\\u003e \\u003c/span\\u003e \\u003cspan\\u003e \\u003cli\\u003e \\u003cp\\u003eDoes the composition of invertebrates vary between grassland locations?\\u003c/p\\u003e \\u003c/li\\u003e \\u003c/span\\u003e \\u003c/ol\\u003e \\u003cp\\u003e\\u003c/p\\u003e \"},{\"header\":\"Methods\",\"content\":\"\\u003cp\\u003eOur research took place in the temperate grasslands of southeast Australia, approximately 50 km northwest of Melbourne. We used three grassland sites within a 15 km radius. The sites are named by location, and all are temperate grassland, varying in vegetation structure, aspect and management. Ingliston was located on a 25-metre roadside strip of remnant grassland that consisted of short \\u003cem\\u003eThemeda triandra\\u003c/em\\u003e amongst an array of forbs and herbs. Myrniong was a grazing exclusion plot with dense swards of \\u003cem\\u003eThemeda triandra.\\u003c/em\\u003e Parwan was a low rainfall plains grassland site on private property that receives ecological burns to reduce biomass, with tussocks of \\u003cem\\u003eAustrostipa, Rytidosperma, Themeda\\u003c/em\\u003e and other grasses and forbs.\\u003c/p\\u003e\\u003cp\\u003eOur experiment involved deploying 15 x 15 cm mesh bags (constructed \\u003cem\\u003esensu\\u003c/em\\u003e Butler et al. \\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e; see Supplementary Figure \\u003cspan refid=\\\"MOESM1\\\" class=\\\"InternalRef\\\"\\u003eS1\\u003c/span\\u003e) containing different grass species to aid in sampling the invertebrate community that colonises the detached grass litter. Mesh bags provide a means to observe natural processes such as litter breakdown and decomposition (Kitz et al. \\u003cspan citationid=\\\"CR46\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e; Schädler \\u0026amp; Brandl \\u003cspan citationid=\\\"CR73\\\" class=\\\"CitationRef\\\"\\u003e2005\\u003c/span\\u003e; Wall et al. \\u003cspan citationid=\\\"CR80\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e), and the aperture size of the bags determines the size classes of invertebrates that can access the litter they contain (Knops et al. \\u003cspan citationid=\\\"CR47\\\" class=\\\"CitationRef\\\"\\u003e2001\\u003c/span\\u003e; Milcu \\u0026amp; Manning \\u003cspan citationid=\\\"CR53\\\" class=\\\"CitationRef\\\"\\u003e2011\\u003c/span\\u003e). Hitherto, grassland studies with litterbags have focussed on the decomposition of litter. This study is the first recorded instance (to the author’s knowledge) of using litter bags to sample invertebrates present.\\u003c/p\\u003e\\u003cp\\u003eWe chose four grass species, two native to the grassland study sites (\\u003cem\\u003eThemeda triandra\\u003c/em\\u003e Forssk., \\u003cem\\u003ePoa labillardierei\\u003c/em\\u003e Steud.\\u003cem\\u003e)\\u003c/em\\u003e and two locally abundant introduced and naturalised species (\\u003cem\\u003eDactylis glomerata\\u003c/em\\u003e L., \\u003cem\\u003eHolcus lanatus\\u003c/em\\u003e L.\\u003cem\\u003e)\\u003c/em\\u003e. We harvested the grass near the Myrniong and Ingliston sites. We used secateurs to cut blades approximately five centimetres from the ground’s surface. We then removed the culms to a practicable degree before bagging samples so that most collected samples comprised leaf blades. We then refrigerated the grass collections at 4ºC until needed. We prepared each mesh bag for deployment by adding 10 g of air-dried grass to each bag. We added a plastic ‘pizza table’ to each bag to maintain airflow and prevent compaction, and we stapled the bags shut.\\u003c/p\\u003e\\u003cp\\u003eWe deployed 24 grass-filled mesh bags in each grassland on 9th November 2022, including six replicates of each of the four species per site. Half of the bags for each species used small-aperture mesh (2 mm) and half used large-aperture mesh (5 mm). When deploying each bag we trimmed the existing growth to ground-level to allow contact of the bag with the soil surface. Each bag was secured to the ground with a metal pin. A sturdy mesh cage was secured over groups of four bags to prevent disturbance from animals (see Supplementary Figure \\u003cspan refid=\\\"MOESM1\\\" class=\\\"InternalRef\\\"\\u003eS1\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003e We collected the bags on 8th December 2022, a month after deployment, using a small serving board under each bag to prevent loss of invertebrates when transferring litter bags to a paper bag. We used a Tullgren-Funnel apparatus in a glasshouse at Federation University, Mount Helen, to extract invertebrates from the grass samples (See Supplementary Figure \\u003cspan refid=\\\"MOESM1\\\" class=\\\"InternalRef\\\"\\u003eS1\\u003c/span\\u003e). We placed the litterbags atop a zinc plate on top of the funnels. Invertebrates fell through the zinc plate as they moved away from light and were collected in a specimen container with 30 mL ethanol affixed at the bottom of the funnel. We allowed five days for this process to occur, with the alcohol levels checked after two days. We then removed the specimen jars from the funnels and stored them upright with complete lids.\\u003c/p\\u003e\\u003cp\\u003eWe used a Nikon SMZ 745 stereomicroscope with a 10–40 x magnification range for identification. We then transferred each sample to a Petri dish via pipette and tallied individuals along the sections of a lined grid. A clicker counter was used for larger samples. We recounted the first five samples at the end to ensure methodology had not evolved, but no significant differences were noted. Specimens were identified to order using Zborowski and Storey (\\u003cspan citationid=\\\"CR85\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e), Daley and Ellingsen (\\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e) and Gooderham and Tsyrlin (\\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e2002\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003eData was compiled in a Microsoft Excel spreadsheet. See Supplementary Data 2 for raw data. All data analyses and figure production were conducted in R (R Core Team \\u003cspan citationid=\\\"CR66\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e) except where stated. We constructed non-metric multidimensional scaling (nMDS) ordination diagrams using the vegan package (Oksanen et al. \\u003cspan citationid=\\\"CR60\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e) based on a square-root transformation of the ordinal abundance data and a Bray-Curtis similarity matrix. We used ellipses projected onto the nMDS plots that grouped sites according to the sample variables (grass species, grass origin, mesh size and site) to observe how they influenced invertebrate composition. Using the statistical software PRIMER (Clarke \\u0026amp; Gorley \\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e2013\\u003c/span\\u003e), we tested for the influence of sample variables on invertebrate composition using analysis of similarity (ANOSIM) and similarity percentage (SIMPER) partitions (Clarke \\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e1993\\u003c/span\\u003e). PRIMER was also used to calculate the following univariate diversity indices for each site: Pielou’s evenness, Shannon and Simpson’s diversity indices, and Margalef richness. Box-and-whisker plots were constructed using the package ggpubr (Kassambara \\u003cspan citationid=\\\"CR44\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e) to visualise the differences in diversity indices among the sites. Non-parametric Kruskal-Wallis tests were used to test for differences in diversity indices among sites, as the data failed the Shapiro-Wilk normality tests.\\u003c/p\\u003e\"},{\"header\":\"Results\",\"content\":\"\\u003cp\\u003eWe sampled a total of 7,933 individual invertebrates from the three sites. These invertebrates belonged to twelve different orders of insects and insect allies (Table\\u0026nbsp;1). The number of captured invertebrates varied from 15 to 535 individuals per litter bag, with a mean of 110\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;10.1. The overall abundance of invertebrates differed significantly among locations (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003ea), but the ordinal richness was similar among the grassland sites, with the sample means of 6.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.3 for Ingliston, 7.8\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.4 for Myrniong, and 6.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.3 for Parwan. Evenness among sites varied significantly and was highest at Myrniong (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003ec). We excluded the larvae present from further analysis. The most abundant groups collected were Poduromorpha, Acari and Thysanoptera, with a combination of these three groups being the most abundant at each location. Acari numbers were similar between the Ingliston and Parwan locations (Table\\u0026nbsp;1). Poduromorpha were most abundant at Parwan, accounting for two-thirds of the total invertebrates recorded from the site (Table\\u0026nbsp;1). Notably, we found that Psocoptera were absent from Parwan, and no Araneae were recorded at Ingliston. There was a significant difference in the invertebrate composition among grass species and grass origin (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eb and \\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003ec, respectively). Coleoptera \\u0026amp; Diplopoda were present in exotic grasses in greater numbers. Conversely, Thysanoptera, Symphypleona and Entobryomorpha appeared in native grasses more frequently.\\u003c/p\\u003e \\u003cp\\u003e \\u003cb\\u003eTable\\u0026nbsp;1.\\u003c/b\\u003e Mean abundance (\\u0026plusmn;\\u0026thinsp;s.e.) of 12 invertebrate taxa, grouped by site and grass species.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eThere was no significant difference in invertebrate composition between the small and large mesh bags (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003ea; Global R: 0.019; Pseudo p\\u0026thinsp;=\\u0026thinsp;0.133). Figure\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eb illustrates the similarity of invertebrate composition based on the species of grass used in the mesh litterbags and shows that \\u003cem\\u003eThemeda\\u003c/em\\u003e litter bags are more tightly grouped than other grass species, suggesting less variability in invertebrate composition. ANOSIM results provide evidence for a significant difference in invertebrate composition among the grass species (Global R\\u0026thinsp;=\\u0026thinsp;0.134, Pseudo p\\u0026thinsp;=\\u0026thinsp;0.001). In Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003ec, the overlap of invertebrate composition between native and exotic grasses is clear. Nevertheless, the ANOSIM results provide evidence for a significant difference in invertebrate composition between the native and exotic grasses (Global R\\u0026thinsp;=\\u0026thinsp;0.091, Pseudo p\\u0026thinsp;=\\u0026thinsp;0.002). A clear grouping is evident among the three sites (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003ed), indicating that the site has the most substantial influence on the invertebrate composition (Global R\\u0026thinsp;=\\u0026thinsp;0.491; pseudo p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.001). The Ingliston samples were more tightly clustered than the other two sites, suggesting greater similarity in invertebrate composition among the samples. SIMPER analysis (Table\\u0026nbsp;2) revealed that Acari and Poduromorpha contributed the most to the dissimilarity between the Myrniong and Ingliston locations. Poduromorpha contributed the most dissimilarity between Myrniong and Parwan, and Ingliston and Parwan.\\u003c/p\\u003e \\u003cp\\u003e \\u003cb\\u003eTable\\u0026nbsp;2\\u003c/b\\u003e. Similarity percentage (SIMPER) partitions comparing taxa dissimilarity among sites.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eAs the site was the most significant driver of invertebrate assemblages, the differences between the sites were further explored with abundance and ordinal diversity indices (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e). The mean invertebrate abundance at Parwan was significantly greater than the other two locations (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003ea), driven by several high-abundance litter bags. In contrast, Myrniong had significantly greater species diversity, evenness, and Margalef richness than the other two sites (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e, p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.001 for each).\\u003c/p\\u003e\"},{\"header\":\"Discussion\",\"content\":\"\\u003cp\\u003eOur study showed that in temperate grassland there is a diverse suite of invertebrate orders that will readily associate with detached grass litter. We found that the invertebrate community was most strongly influenced by the study location, suggesting there is an important level of spatial variation among litter-dwelling communities that may be driven by climate and abiotic differences among sites. Invertebrate community composition was also affected to a lesser extent by the species of grass and its origin but was not affected by the gauge of mesh bags used to collect samples.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cem\\u003eGrassland location\\u003c/em\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eWe found that the sampling location had the strongest effect on invertebrate community composition. The literature suggests that vegetation structure (New 2000; Schultz et al. 2017), landscape context\\u0026nbsp;(Barton et al. 2024; Oliver et al. 2006), management history (Abraham \\u0026amp; Morgan 2018; Bromham et al. 1999) and climate (Barnett \\u0026amp; Facey 2016; Garcia-Palacios et al. 2013) may all exude a degree of influence on shaping invertebrate habitat. Several studies have suggested the sward structure or microhabitat influences the suitability for grassland fauna (Barton et al. 2024; Lindsay \\u0026amp; Cunningham 2009; Price et al. 2019). Our three locations varied considerably in sward structure and available niches. Myrniong had a closed structure with dense \\u003cem\\u003eThemeda\\u003c/em\\u003e growth, providing a sheltered aspect on a slight slope with good drainage. Ingliston had a short semi-open structure of grass and forbs, which was seasonally waterlogged in moist depressions. Parwan had an open sward structure, providing thermophilic habitat niches. Small forbs and rocks amongst tussocks added a layer of habitat complexity. These differences may explain the variation in invertebrate communities recorded. It would be useful to test for differences in litter-dwelling invertebrates among a larger range of sites that span a broad range of site variables to directly test these impacts.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cem\\u003eGrass origin\\u003c/em\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe influence of native and exotic grass species on litter invertebrate community composition was significant in this study. This result was driven by the preferences of a select few orders: Coleoptera \\u0026amp; Diplopoda were more frequently captured in exotic grass. Thysanoptera, Symphypleona and Entobryomorpha appeared in native grasses in higher\\u0026nbsp;numbers. As only five of the twelve orders demonstrated a preference towards grass origin, this result suggests generalists and specialists in the invertebrate fauna. Similar invertebrate responses in alien and native grass species have been noted elsewhere, albeit within the context of alien plant invasions (Belnap \\u0026amp; Phillips 2001; Litt \\u0026amp; Steidl 2010). Local observations note that exotic grass invasions can lead to a homogenised structure, and decreased habitat diversity (New 2019), whereas overall invertebrate abundance had a positive association with native grass cover in Australia\\u0026rsquo;s grassy woodlands (Lindsay \\u0026amp; Cunningham 2009).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cem\\u003eGrass species\\u003c/em\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eWe found a significant difference in the invertebrates captured from the four grass species. Invertebrate responses to single grass species are poorly understood, at least locally. One Australian grassland study noted that some invertebrate species vacuum-sampled from live \\u003cem\\u003eThemeda triandra\\u003c/em\\u003e and \\u003cem\\u003ePoa labillardierei\\u003c/em\\u003e tussocks exhibited strong grass species preferences (Reid \\u0026amp; Hochuli 2007). Plant traits such as nutrient status that influence litter variables can influence decomposition rate (Bradford et al. 2016; Cornwell et al. 2008). Australia\\u0026rsquo;s native grasses have evolved for millennia in isolation (Bryceson \\u0026amp; Morgan 2022) with dry climate, nutrient-poor soils, and frequent fire leading to a distinct assemblage of native flora (Orians \\u0026amp; Milewski 2007). High lignin and low nitrogen content result in a recalcitrant litter that does not appear to follow the same decomposition trajectory as its northern hemisphere counterparts (Butler et al. 2023). \\u003cem\\u003eThemeda\\u003c/em\\u003e and \\u003cem\\u003ePoa\\u003c/em\\u003e differ in chemical composition, with Butler et al. (2023) demonstrating that \\u003cem\\u003eThemeda\\u003c/em\\u003e decomposed at a greater rate than \\u003cem\\u003ePoa\\u0026nbsp;\\u003c/em\\u003eover a 39-week period (Butler et al. 2023). Functional traits and nutrient status may have affected their suitability for invertebrate decomposers.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003eThysanoptera and Collembola (Entobryomorpha, Symphypleona and Poduromorpha) are two groups that have exhibited specialist plant associations, and this may have influenced our results. The endemic grass-dwelling members of Thysanoptera have been noted to have an association with Australian native grasses, including \\u003cem\\u003eThemeda\\u0026nbsp;\\u003c/em\\u003eand \\u003cem\\u003ePoa\\u003c/em\\u003e species, whereas exotic Thysanoptera taxa were associated with exotic grasses (Mound 2011). In Australia, Collembola has a high rate of endemism (Greenslade 2007), with exotic Collembola used as an indicator of disturbance. Exotic Collembola are largely absent from intact, low-nutrient, environments, suggesting disturbance and landscape change facilitate invasion (Greenslade 2018). Specialist fauna that has co-adapted to Australian flora may have impacted the invertebrates\\u0026rsquo; preferences for grass species within the study.\\u003c/p\\u003e\\n\\u003cp\\u003eThe photosynthetic pathways of the grasses may also have influenced the invertebrate communities observed. Our study species were all C3 grasses except for \\u003cem\\u003eThemeda\\u003c/em\\u003e, which is a C4 grass. We observed that \\u003cem\\u003eThemeda\\u0026nbsp;\\u003c/em\\u003eshowed lower variability in the invertebrate community among samples than the other grasses. This may be because C4 plants are generally thought to provide poorer food quality to decomposers due to low nutrient and high fibre content (Barbehenn \\u0026amp; Bernays 1992; Scheirs et al. 2001; Scheunemann et al. 2010).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cem\\u003eMesh size\\u003c/em\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe use of two different mesh sizes in litter bags did not influence the invertebrate community sampled. Comparing our use of litterbags with other studies is challenging as previous research has focused on the disappearance rate of plant material rather than invertebrates associated with this process. While some studies have suggested that using a smaller mesh size excludes larger invertebrates (Delgado-Baquerizo et al. 2020; Handa et al. 2014; Kitz et al. 2015; Makkonen et al. 2012; Milcu \\u0026amp; Manning 2011) and leads to slower decomposition rates, results from other studies (Cates et al. 2021; Sch\\u0026auml;dler \\u0026amp; Brandl 2005; Wall et al. 2008) make it difficult to draw definitive conclusions. Kampichler and Bruckner (2009) noted that comparing studies is challenging due to variations in mesh aperture, bag fillings, and the specific aims of the research. Our results suggest that the 2-mm mesh (which was easier to work with than the larger mesh size) was sufficient for observing the invertebrate community associated with decomposing grass litter that were the focus of this study.\\u003c/p\\u003e\"},{\"header\":\"Conclusions\",\"content\":\"\\u003cp\\u003eOur study shows that the invertebrate community associated with grass litter is influenced by grass identity and grass origin. However, this influence is slight when compared to the differences emerging from separate locations. The differential response to the addition of exotic grass suggests that specialist invertebrates are present in the grass litter communities, and the encroachment of weeds may threaten the local persistence of invertebrates such as members of Entobryomorpha, Symphypleona and Thysanoptera orders, which appear to favour native grasses.\\u003c/p\\u003e \\u003cp\\u003eThe community variation among locations, even at a basic taxonomic level, emphasises the need for greater recognition of the role that invertebrates play in grassland ecosystems. Strategies aiming to improve grassland condition, such as maintaining structural diversity and managing grazing levels, generally benefit grassland invertebrates. However, our study shows significant variability within sites, making it challenging to assess these benefits at specific locations. Therefore, improving monitoring of grassland invertebrates will help us measure the outcomes of management practices. In turn, this will allow us to anticipate how grasslands might respond to global change and develop strategies for conserving invertebrate diversity.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\" \\u003cp\\u003e \\u003cstrong\\u003eConflict of Interest.\\u003c/strong\\u003e \\u003cp\\u003eThe authors declare no conflict of interest.\\u003c/p\\u003e \\u003c/p\\u003e\\u003ch2\\u003eAuthor Contribution\\u003c/h2\\u003e\\u003cp\\u003eAll authors contributed to the study conception and design, material preparation, data collection and analysis. AMW wrote the original draft manuscript and prepared table 2. NLS prepared figures 1-2. All authors reviewed and edited the manuscript and approved the final manuscript.\\u003c/p\\u003e\\u003ch2\\u003eAcknowledgement\\u003c/h2\\u003e\\u003cp\\u003eAllison would like to acknowledge the memory of Roslynn Shearer, whose encouragement, friendship, and support contributed to the fruition of this research. Thanks to Tom Mills for assistance with fieldwork, and Penelope Greenslade for input into initial discussions. We are grateful to Simon \\u0026amp; Lorraine Jolly for allowing the use of their property. Thanks to Tom Miller and the Moorabool Shire for allowing the use of further study locations. Finally, we acknowledge and pay our respects to the Wadawurrung and Wurundjeri Traditional Owners of the land on which this research took place.\\u003c/p\\u003e\\u003ch2\\u003eData Availability\\u003c/h2\\u003e\\u003cp\\u003eData is provided within the supplementary information files.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eAbraham J, Morgan JW (2018) Effects of time-since-fire on the invertebrate communities of Kangaroo grass \\u003cem\\u003e\\u0026ldquo;Themeda\\u003c/em\\u003e \\u003cem\\u003etriandra\\u0026rdquo;\\u003c/em\\u003e-dominated grasslands in Melbourne, Victoria. The Vic Naturalist 135:36-46. https://search.informit.org/doi/10.3316/informit.350603692010878 \\u003c/li\\u003e\\n\\u003cli\\u003eAerts R (1997) Climate, leaf litter chemistry and leaf litter decomposition in terrestrial ecosystems: a triangular relationship. Oikos 79:439-449. https://doi.org/10.2307/3546886 \\u003c/li\\u003e\\n\\u003cli\\u003eAntos M, Williams N (2015) The wildlife of our grassy landscape. In: N Williams, A Marshall \\u0026amp; J Morgan (eds), Land of Sweeping Plains: Managing and Restoring the Native Grasslands of south-eastern Australia. CSIRO Publishing, Melbourne, pp 87-114\\u003c/li\\u003e\\n\\u003cli\\u003eAustin AT, Vivanco L (2006) Plant litter decomposition in a semi-arid ecosystem controlled by photodegradation. Nat 442:555-558. https://doi.org/10.1038/nature05038\\u003c/li\\u003e\\n\\u003cli\\u003eBagyaraj DJ, Nethravathi CJ, Nitin KS (2016) Soil Biodiversity and Arthropods: Role in Soil Fertility. In: AK Chakravarthy \\u0026amp; S Sridhara (eds), Economic and Ecological Significance of Arthropods in Diversified Ecosystems: Sustaining Regulatory Mechanisms. Springer Singapore, Singapore, pp 17-51\\u003c/li\\u003e\\n\\u003cli\\u003eBarbehenn RV, Bernays EA (1992) Relative nutritional quality of C 3 and C 4 grasses for a graminivorous lepidopteran, Paratrytone melane (Hesperiidae). Oecologia 92:97-103. https://doi.org/10.1007/BF00317268 \\u003c/li\\u003e\\n\\u003cli\\u003eBarnett KL, Facey SL (2016) Grasslands, Invertebrates, and Precipitation: A Review of the Effects of Climate Change. Front Plant Sci 7:1196. https://doi.org/10.3389/fpls.2016.01196 \\u003c/li\\u003e\\n\\u003cli\\u003eBarton PS, Evans MJ (2017) Insect biodiversity meets ecosystem function: differential effects of habitat and insects on carrion decomposition. Ecol Entomol 42:364-74. https://doi.org/10.1111/een.12395 \\u003c/li\\u003e\\n\\u003cli\\u003eBarton PS, Evans MJ, Lewis J (2024) Microhabitats shape ant community structure in a spatially heterogeneous grassy woodland. Ecosphere 15:4798. https://doi.org/10.1002/ecs2.4798 \\u003c/li\\u003e\\n\\u003cli\\u003eBelnap J, Phillips SL (2001) Soil biota in an ungrazed grassland: response to annual grass (\\u003cem\\u003eBromus tectorum\\u003c/em\\u003e) invasion. Ecol Appl 11:1261-75. https://doi.org/10.1890/1051-0761(2001)011[1261:SBIAUG]2.0.CO;2 \\u003c/li\\u003e\\n\\u003cli\\u003eBradford MA, Berg B, Maynard DS, Wieder WR, Wood, SA (2016) Future Directions: Understanding the dominant controls on litter decomposition. J Ecol 104:229-38. http://www.jstor.org/stable/24762886 \\u003c/li\\u003e\\n\\u003cli\\u003eBromham L, Cardillo M, Bennett AF, Elgar, MA (1999) Effects of stock grazing on the ground invertebrate fauna of woodland remnants. Aust J Ecol 24:199-207. https://doi.org/10.1046/j.1442-9993.1999.00963.x \\u003c/li\\u003e\\n\\u003cli\\u003eBryceson SR, Morgan JW (2022) The Australasian grass flora in a global context. J Syst Evol 60:675-690. https://doi.org/10.1111/jse.12839 \\u003c/li\\u003e\\n\\u003cli\\u003eButler F, Good M, Morgan J, Schultz N (2023) Relative contribution of photodegradation to litter breakdown in Australian grasslands. Ecol Evol 13:10710. https://doi.org/10.1002/ece3.10710 \\u003c/li\\u003e\\n\\u003cli\\u003eCardoso P, Erwin TL, Borges PA, New TR (2011) The seven impediments in invertebrate conservation and how to overcome them. Biol Conserv 144:2647-2655. https://doi.org/10.1016/j.biocon.2011.07.024 \\u003c/li\\u003e\\n\\u003cli\\u003eCates AM, Wills BD, Kim TN, Landis DA, Gratton C, Read HW, Jackson RD (2021) No evidence of top-down effects by ants on litter decomposition in a temperate grassland. Ecosphere 12:03638. https://doi.org/10.1002/ecs2.3638 \\u003c/li\\u003e\\n\\u003cli\\u003eChapman AD (2009) Numbers of living species in Australia and the World, 2nd edn. Department of the Environment, Water, Heritage and the Arts, Australian Government, Canberra.\\u003c/li\\u003e\\n\\u003cli\\u003eClarke K, Gorley R (2013) Primer 6: Version 6.1.16. User Manual/Tutorial. Primer-E Ltd, Plymouth\\u003c/li\\u003e\\n\\u003cli\\u003eClarke KR (1993) Non‐parametric multivariate analyses of changes in community structure. Aust J Ecol 181:117-43. https://doi.org/10.1111/j.1442-9993.1993.tb00438.x \\u003c/li\\u003e\\n\\u003cli\\u003eClay RE (2014) Potential Effects of the Loss of Native Grasses on Grassland Invertebrate Diversity in Southeastern Australia. Int J Ecol 2014:202056. https://doi.org/10.1155/2014/202056\\u003c/li\\u003e\\n\\u003cli\\u003eColeman DC, Callaham MA, Crossley DA (2017) Fundamentals of Soil Ecology, 3rd edn. Academic Press, London.\\u003c/li\\u003e\\n\\u003cli\\u003eCornwell WK, Cornelissen JH, Amatangelo K, Dorrepaal E, Eviner VT, Godoy O, Hobbie SE, Hoorens B, Kurokawa H, Perez-Harguindeguy N (2008) Plant species traits are the predominant control on litter decomposition rates within biomes worldwide, Ecol Lett 11:1065-1071. https://doi.org/10.1111/j.1461-0248.2008.01219.x \\u003c/li\\u003e\\n\\u003cli\\u003eCuevas E, Medina E (1988) Nutrient dynamics within amazonian forests: II. Fine root growth, nutrient availability and leaf litter decomposition. Oecologia 76:222-235. https://doi.org/10.1007/BF00379956\\u003c/li\\u003e\\n\\u003cli\\u003eCulliney TW (2013) Role of Arthropods in Maintaining Soil Fertility. Agriculture 3:629-659 https://doi.org/10.3390/agriculture3040629\\u003c/li\\u003e\\n\\u003cli\\u003eCurry J (1987) The invertebrate fauna of grassland and its influence on productivity. III. Effects on soil fertility and plant growth. Grass Forage Sci 42:325-341 https://doi.org/10.1111/j.1365-2494.1987.tb02121.x\\u003c/li\\u003e\\n\\u003cli\\u003eDaley A, Ellingsen K (2012) Insects of Tasmania: An online field guide. https://tasmanianinsectfieldguide.com/. Accessed 25 April 2023\\u003c/li\\u003e\\n\\u003cli\\u003eDeca\\u0026euml;ns T (2010) Macroecological patterns in soil communities. Global Ecol Biogeogr 19:287-302. https://doi.org/10.1111/j.1466-8238.2009.00517.x\\u003c/li\\u003e\\n\\u003cli\\u003eDelgado-Baquerizo M, Reich PB, Trivedi C, Eldridge DJ, Abades S, Alfaro FD, Bastida F, Berhe AA, Cutler NA, Gallardo A, Garc\\u0026iacute;a-Vel\\u0026aacute;zquez L, Hart SC, Hayes PE, He J-Z, Hseu Z-Y, Hu H-W, Kirchmair M, Neuhauser S, P\\u0026eacute;rez CA, Reed SC, Santos F, Sullivan BW, Trivedi P, Wang J-T, Weber-Grullon L, Williams MA, Singh BK (2020) Multiple elements of soil biodiversity drive ecosystem functions across biomes. Nat Ecol Evol 4:210-20. https://doi.org/10.1038/s41559-019-1084-y\\u003c/li\\u003e\\n\\u003cli\\u003eFay PA, Prober SM, Harpole WS, Knops JMH, Bakker JD, Borer ET, Lind EM, MacDougall AS, Seabloom EW, Wragg PD, Adler PB, Blumenthal DM, Buckley YM, Chu C, Cleland EE, Collins SL, Davies KF, Du G, Feng X, Firn J, Gruner DS, Hagenah N, Hautier Y, Heckman RW, Jin VL, Kirkman KP, Klein J, Ladwig LM, Li Q, McCulley RL, Melbourne BA, Mitchell CE, Moore JL, Morgan JW, Risch AC, Sch\\u0026uuml;tz M, Stevens CJ, Wedin DA, Yang LH (2015) Grassland productivity limited by multiple nutrients. Nat Plants 1:15080. http://dx.doi.org/10.1038/nplants.2015.80 \\u003c/li\\u003e\\n\\u003cli\\u003eGarcia-Palacios P, Maestre FT, Kattge J, Wall DH (2013) Climate and litter quality differently modulate the effects of soil fauna on litter decomposition across biomes. Ecol Lett 16:1045-1053. https://doi.org/10.1111/ele.12137 \\u003c/li\\u003e\\n\\u003cli\\u003eGessner MO, Swan CM, Dang CK, McKie BG, Bardgett RD, Wall DH, H\\u0026auml;ttenschwiler S (2010) Diversity meets decomposition. Trends Ecol Evol 25:372-880. https://doi.org/10.1016/j.tree.2010.01.010 \\u003c/li\\u003e\\n\\u003cli\\u003eGibson L, New T (2007) Characterising insect diversity on Australia\\u0026rsquo;s remnant native grasslands: ants (Hymenoptera: Formicidae) and beetles (Coleoptera) at Craigieburn Grasslands Reserve, Victoria. J Insect Conserv 11:409-413. https://doi.org/10.1007/s10841-006-9051-8 \\u003c/li\\u003e\\n\\u003cli\\u003eGongalsky KB (2021) Soil macrofauna: Study problems and perspectives. Soil Biol Biochem 159:108281. https://doi.org/10.1016/j.soilbio.2021.108281\\u003c/li\\u003e\\n\\u003cli\\u003eGooderham J, Tsyrlin E (2002) The Waterbug Book: A Guide to the Freshwater Macroinvertebrates of Temperate Australia. CSIRO Publishing, Melbourne.\\u003c/li\\u003e\\n\\u003cli\\u003eGreenslade P (1997) Short term effects of a prescribed burn on invertebrates in grassy woodland in southeastern Australia. Memoirs of Museum Victoria 56:305-312. https://doi.org/10.24199/j.mmv.1997.56.18 \\u003c/li\\u003e\\n\\u003cli\\u003eGreenslade P (2007) The potential of Collembola to act as indicators of landscape stress in Australia. Aust J Exp Agr 47: 424-34. https://doi.org/10.1071/EA05264\\u003c/li\\u003e\\n\\u003cli\\u003eGreenslade P (2018) Why are there so many exotic Springtails in Australia? A review. Soil organisms 90:141-156. https://doi.org/10.25674/y9tz-1d49\\u003c/li\\u003e\\n\\u003cli\\u003eGriffiths HM, Ashton LA, Parr CL, Eggleton P (2021) The impact of invertebrate decomposers on plants and soil. New Phytol 231:2142-2149. https://doi.org/10.1111/nph.17553\\u003c/li\\u003e\\n\\u003cli\\u003eHanda IT, Aerts R, Berendse F, Berg MP, Bruder A, Butenschoen O, Chauvet E, Gessner MO, Jabiol J, Makkonen, M (2014) Consequences of biodiversity loss for litter decomposition across biomes. Nat 509:218-221. https://doi.org/10.1038/nature13247\\u003c/li\\u003e\\n\\u003cli\\u003eH\\u0026auml;ttenschwiler S, Tiunov AV, Scheu S (2005) Biodiversity and Litter Decomposition in Terrestrial Ecosystems. Annu Rev Ecol Evol S 36:191-218. https://doi.org/10.1146/annurev.ecolsys.36.112904.151932\\u003c/li\\u003e\\n\\u003cli\\u003eHeděnec P, Jim\\u0026eacute;nez JJ, Moradi J, Domene X, Hackenberger D, Barot S, Frossard A, Oktaba L, Filser, J, Kindlmann P (2022) Global distribution of soil fauna functional groups and their estimated litter consumption across biomes. Sci Rep 12:17362. https://doi.org/10.1038/s41598-022-21563-z \\u003c/li\\u003e\\n\\u003cli\\u003eHungate BA, Barbier EB, Ando AW, Marks SP, Reich PB, Van Gestel N, Tilman D, Knops JM, Hooper DU, Butterfield, BJ (2017) The economic value of grassland species for carbon storage. Sci Advances 3:1601880 DOI: 10.1126/sciadv.1601880\\u003c/li\\u003e\\n\\u003cli\\u003eKampichler C, Bruckner A (2009) The role of microarthropods in terrestrial decomposition: a meta‐analysis of 40 years of litterbag studies. Biol Rev 84:375-389. https://doi.org/10.1111/j.1469-185X.2009.00078.x\\u003c/li\\u003e\\n\\u003cli\\u003eKassambara A (2023) ggpubr: \\u0026apos;ggplot2\\u0026apos; Based Publication Ready Plots, R package version 0.6.0, https://rpkgs.datanovia.com/ggpubr/ Accessed 8 May 2023\\u003c/li\\u003e\\n\\u003cli\\u003eKing KL, Hutchinson KJ (1983) The effects of sheep grazing on invertebrate numbers and biomass in unfertilized natural pastures of the New England Tablelands (NSW). Aust J Ecol 8: 245-255. https://doi.org/10.1111/j.1442-9993.1983.tb01322.x \\u003c/li\\u003e\\n\\u003cli\\u003eKitz F, Steinwandter M, Traugott M, Seeber J (2015) Increased decomposer diversity accelerates and potentially stabilises litter decomposition. Soil Biol Biochem 83:138-141. https://doi.org/10.1016/j.soilbio.2015.01.026 \\u003c/li\\u003e\\n\\u003cli\\u003eKnops JM, Wedin D, Tilman D (2001) Biodiversity and decomposition in experimental grassland ecosystems. Oecologia 126:429-433. https://doi.org/10.1007/s004420000537 \\u003c/li\\u003e\\n\\u003cli\\u003eLavelle P, Deca\\u0026euml;ns T, Aubert M, Barot Sb, Blouin M, Bureau F, Margerie P, Mora P, Rossi, J-P (2006) Soil invertebrates and ecosystem services. Eur J Soil Biol 42:S3-S15. https://doi.org/10.1016/j.ejsobi.2006.10.002 \\u003c/li\\u003e\\n\\u003cli\\u003eLindsay EA, Cunningham SA (2009) Livestock grazing exclusion and microhabitat variation affect invertebrates and litter decomposition rates in woodland remnants. Forest Ecol Manag 258:178-187. https://doi.org/10.1016/j.foreco.2009.04.005 \\u003c/li\\u003e\\n\\u003cli\\u003eLitt AR, Steidl RJ (2010) Insect assemblages change along a gradient of invasion by a nonnative grass. Biol Invasions 12:3449-3463. https://doi.org/10.1007/s10530-010-9743-6 \\u003c/li\\u003e\\n\\u003cli\\u003eMakkonen M, Berg MP, Handa IT, H\\u0026auml;ttenschwiler S, van Ruijven J, van Bodegom PM, Aerts R (2012) Highly consistent effects of plant litter identity and functional traits on decomposition across a latitudinal gradient. Ecol Lett 15:1033-1041. https://doi.org/10.1111/j.1461-0248.2012.01826.x \\u003c/li\\u003e\\n\\u003cli\\u003eMarshall D, Fitzsimons JA (2008) Challenges for native grassland conservation on Victoria\\u0026apos;s Northern Plains. Australas Plant Conserv 16:24-5. https://search.informit.org/doi/10.3316/informit.023279248905647 \\u003c/li\\u003e\\n\\u003cli\\u003eMilcu A, Manning P (2011) All size classes of soil fauna and litter quality control the acceleration of litter decay in its home environment. Oikos 120:1366-1370. https://doi.org/10.1111/j.1600-0706.2010.19418.x \\u003c/li\\u003e\\n\\u003cli\\u003eMiller L, New T (1997) Mount Piper grasslands: pitfall trapping of ants and interpretation of habitat variability. Memoirs of the Museum of Victoria 56:377-381\\u003c/li\\u003e\\n\\u003cli\\u003eMorgan JW (1998) Patterns of invasion of an urban remnant of a species-rich grassland in southeastern Australia by non-native plant species. J Veg Sci 9:181-190. https://doi.org/10.2307/3237117 \\u003c/li\\u003e\\n\\u003cli\\u003eMorris MG (2000) The effects of structure and its dynamics on the ecology and conservation of arthropods in British grasslands. Biol Conserv 95:129-142. https://doi.org/10.1016/S0006-3207(00)00028-8 \\u003c/li\\u003e\\n\\u003cli\\u003eMound LA (2011) Grass-dependent Thysanoptera of the family Thripidae from Australia, Zootaxa 3064:1-40. https://doi.org/10.11646/zootaxa.3064.1.1 \\u003c/li\\u003e\\n\\u003cli\\u003eNew TR (2000) How Useful are Ant Assemblages for Monitoring Habitat Disturbance on Grasslands in South Eastern Australia? J Insect Conserv 4:153-159. https://doi.org/10.1023/A:1009668817271 \\u003c/li\\u003e\\n\\u003cli\\u003eNew TR (2019) Insect Conservation and Australia\\u0026rsquo;s Grasslands. Springer Nature, Cham, Switzerland. https://doi.org/10.1007/978-3-030-22780-7 \\u003c/li\\u003e\\n\\u003cli\\u003eOksanen J, Blanchet FG, Friendly M, Kindt R, Legendre P, McGlinn, D, Minchin PR, O\\u0026rsquo;Hara RB, Simpson GL, Solymos P, Stevens MHH, Szoecs E, Wagner H (2019) Vegan: Community Ecology Package. R Package Version 2.5\\u0026ndash;6. https://CRAN.R-project.org/package=vegan.\\u003c/li\\u003e\\n\\u003cli\\u003eOliver I, Pearce S, Greenslade PJ \\u0026amp; Britton, DR (2006) Contribution of paddock trees to the conservation of terrestrial invertebrate biodiversity within grazed native pastures. Austral Ecol 31:1-12. https://doi.org/10.1111/j.1442-9993.2006.01537.x\\u003c/li\\u003e\\n\\u003cli\\u003eOrgiazzi A, Bardgett R, Barrios E, Behan-Pelletier V, Briones M, Chotte J, De Deyn G, Eggleton P, Fierer N, Fraser T, Hedlund K, Jeffery S, Johnson N, Jones A, Kandeler E, Kaneko N, Lavelle P, Lemanceau P, Miko L, Montanarella L, Moreira F, Ramirez K, Scheu S, Singh B, Six J, van der Putten W, Wall DE (2016) Global Soil Biodiversity Atlas, Publications Office of the European Union, Luxembourg. \\u003c/li\\u003e\\n\\u003cli\\u003eOrians GH, Milewski AV (2007) Ecology of Australia: the effects of nutrient-poor soils and intense fires. Biol Rev 82:393-423. https://doi.org/10.1111/j.1469-185X.2007.00017.x\\u003c/li\\u003e\\n\\u003cli\\u003ePatoine G, Thakur MP, Friese J, Nock C, H\\u0026ouml;nig L, Haase J, Scherer-Lorenzen M, Eisenhauer N (2017) Plant litter functional diversity effects on litter mass loss depend on the macro-detritivore community. Pedobiologia 65:29-42. https://doi.org/10.1016/j.pedobi.2017.07.003 \\u003c/li\\u003e\\n\\u003cli\\u003ePrice JN, Good MK, Schultz NL, Guja LK, Morgan JW (2019) Multivariate drivers of diversity in temperate Australian native grasslands. Aust J Bot 67:367-380. https://doi.org/10.1071/BT18190\\u003c/li\\u003e\\n\\u003cli\\u003eR Core Team (2023) R: A Language and Environment for Statistical Computing, The R Foundation for Statistical Computing, Vienna, Austria. https://www.r-project.org/\\u003c/li\\u003e\\n\\u003cli\\u003eReid, AM \\u0026amp; Hochuli, DF (2007) Grassland invertebrate assemblages in managed landscapes: Effect of host plant and microhabitat architecture. Austral Ecol 32:708-718. https://doi.org/10.1111/j.1442-9993.2007.01767.x \\u003c/li\\u003e\\n\\u003cli\\u003eRosenberg Y, Bar-On YM, Fromm A, Ostikar M, Shoshany A, Giz O, Milo R (2023) The global biomass and number of terrestrial arthropods. Sci Advances 9:eabq4049 https://doi.org/10.1126/sciadv.abq4049 \\u003c/li\\u003e\\n\\u003cli\\u003eRoss CE, Barton PS, McIntyre S, Cunningham SA, Manning AD (2017) Fine‐scale drivers of beetle diversity are affected by vegetation context and agricultural history. Austral Ecol 42:831-843. https://doi.org/10.1111/aec.12506 \\u003c/li\\u003e\\n\\u003cli\\u003eRoy PG, Delpratt J (2006) Victorian Basalt Plains Grasslands - an Overview. Australas Plant Conserv 15:4-6. https://search.informit.org/doi/10.3316/informit.054675805475714 \\u003c/li\\u003e\\n\\u003cli\\u003eSamways MJ, Caldwell P, Osborn R (1996) Ground-living invertebrate assemblages in native, planted and invasive vegetation in South Africa. Agr Ecosyst Environ 59:19-32. https://doi.org/10.1016/0167-8809(96)01047-X \\u003c/li\\u003e\\n\\u003cli\\u003eSaunders ME, Barton PS, Bickerstaff JRM, Frost L, Latty T, Lessard BD, Lowe EC, Rodriguez J, White TE, Umbers KDL (2021) Limited understanding of bushfire impacts on Australian invertebrates. Insect Conserv Diver 14:285-293. https://doi.org/10.1111/icad.12493 \\u003c/li\\u003e\\n\\u003cli\\u003eSch\\u0026auml;dler M, Brandl R (2005) Do invertebrate decomposers affect the disappearance rate of litter mixtures? Soil Biol Biochem 37:329-337. https://doi.org/10.1016/j.soilbio.2004.07.042 \\u003c/li\\u003e\\n\\u003cli\\u003eScheirs J, De Bruyn L, Verhagen R (2001) A test of the C3\\u0026ndash;C4 hypothesis with two grass miners. Ecol. 82:410-421 https://doi.org/10.1016/j.soilbio.2004.07.042 \\u003c/li\\u003e\\n\\u003cli\\u003eScheunemann N, Scheu S, Butenschoen O (2010) Incorporation of decade old soil carbon into the soil animal food web of an arable system. Appl Soil Ecol 46:59-63. https://doi.org/10.1016/j.apsoil.2010.06.014 \\u003c/li\\u003e\\n\\u003cli\\u003eSchultz N, Keatley M, Antos M, Wong N, Moxham C, Farmilo B, Morgan JW (2017) The golf ball method for rapid assessment of grassland structure. Ecol Manag Restor 18:134-140. https://doi.org/10.1111/emr.12254 \\u003c/li\\u003e\\n\\u003cli\\u003eSolascasas P, Azc\\u0026aacute;rate FM, Hevia V (2022) Edaphic arthropods as indicators of the ecological condition of temperate grassland ecosystems: A systematic review. Ecol Indic 142:109277. https://doi.org/10.1016/j.ecolind.2022.109277 \\u003c/li\\u003e\\n\\u003cli\\u003eSwift MJ, Heal OW, Anderson, JM (1979) Decomposition in terrestrial ecosystems, vol. 5, Blackwell Scientific, Oxford.\\u003c/li\\u003e\\n\\u003cli\\u003eTaylor GS, Braby MF, Moir ML, Harvey MS, Sands DP, New TR, Kitching RL, McQuillan PB, Hogendoorn K, Glatz RV, Andren M, Cook JM, Henry SC, Valenzuela I, Weinstein P (2018) Strategic national approach for improving the conservation management of insects and allied invertebrates in Australia. Austral Entomol 57:124-149. https://doi.org/10.1111/aen.12343 \\u003c/li\\u003e\\n\\u003cli\\u003eWall DH, Bradford MA, St. John MG, Trofymow JA, Behan‐Pelletier V, Bignell DE, Dangerfield JM, Parton WJ, Rusek J, Voigt W (2008) Global decomposition experiment shows soil animal impacts on decomposition are climate‐dependent. Global Change Biol 14:2661-2677. https://doi.org/10.1111/j.1365-2486.2008.01672.x \\u003c/li\\u003e\\n\\u003cli\\u003eWhite RP, Murray S, Rohweder M, Prince S, Thompson K (2000) Grassland Ecosystems, World Resources Institute, Washington, DC, USA.\\u003c/li\\u003e\\n\\u003cli\\u003eYang Y, Tilman D, Furey G, Lehman C (2019) Soil carbon sequestration accelerated by restoration of grassland biodiversity. Nat Commun 10:718. https://doi.org/10.1038/s41467-019-08636-w \\u003c/li\\u003e\\n\\u003cli\\u003eYen A (1999) Grassland invertebrates of the western Victorian basalt plains: plant crunchers or forgotten lunches. In \\u0026lsquo;The Great Plains crash: Proceedings of a conference on the grasslands and grassy woodlands of Victoria\\u0026rsquo;. Victorian Institute of Technology (Ed. RN Jones) pp. 57-68. Indigenous Flora and Fauna Association/ Victorian National Parks Association, East Melbourne, Victoria\\u003c/li\\u003e\\n\\u003cli\\u003eYoshioka A, Kadoya T, Suda S-I, Washitani I (2010) Impacts of weeping lovegrass (\\u003cem\\u003eEragrostis\\u003c/em\\u003e \\u003cem\\u003ecurvula\\u003c/em\\u003e) invasion on native grasshoppers: responses of habitat generalist and specialist species. Biol Invasions 12:531-539. https://doi.org/10.1007/s10530-009-9456-x \\u003c/li\\u003e\\n\\u003cli\\u003eZborowski P, Storey R (2017) A Field Guide to Insects in Australia, 4 edn. New Holland, Sydney.\\u003c/li\\u003e\\n\\u003cli\\u003eZeng X, Gao H, Wang R, Majcher BM, Woon JS, Wenda C, Eggleton P, Griffiths HM, Ashton LA (2024) Global contribution of invertebrates to forest litter decomposition. Ecol Lett 27:e14423. https://doi.org/10.1111/ele.14423 \\u003c/li\\u003e\\n\\u003c/ol\\u003e\"},{\"header\":\"Tables\",\"content\":\"\\u003cp\\u003eTable 1 and 2 are available in the Supplementary Files section.\\u003c/p\\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\":\"info@researchsquare.com\",\"identity\":\"journal-of-insect-conservation\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"jico\",\"sideBox\":\"Learn more about [Journal of Insect Conservation](http://link.springer.com/journal/10841)\",\"snPcode\":\"10841\",\"submissionUrl\":\"https://submission.nature.com/new-submission/10841/3\",\"title\":\"Journal of Insect Conservation\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false},\"keywords\":\"decomposition, grassland ecology, invertebrates, litterbag, Themeda triandra, Poa labillardierei\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-5486939/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-5486939/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eGrasslands comprise a significant portion of terrestrial ecosystems, contributing an estimated 20% of global carbon stores. Biomass is recycled in these systems by photodegradation, biotic decomposition, and through disturbances such as fire or grazing. Yet the diversity of invertebrates and their potential role in biotic decomposition remains unclear in many grasslands worldwide. To help close this knowledge gap we conducted a litter bag experiment to test for the effects of four grass species (two native and two exotic) and two mesh sizes on invertebrate assemblages surveyed at three grassland sites in southeastern Australia. We collected 7,933 invertebrates across twelve arthropod orders and found that all sites had a diverse and abundant invertebrate community that readily interacted with detached grass litter. Study site had the largest effect on invertebrate composition, with significant differences found for Acari, Poduromorpha and Thysanoptera. Grass species identity also had a small but significant effect on invertebrate composition, but there was no effect of litter bag mesh size on the invertebrates.\\u003c/p\\u003e \\u003cp\\u003e \\u003cem\\u003eConservation implications\\u003c/em\\u003e: We found that both geographic and floristic factors were important drivers of variation in grassland invertebrate communities linked to decomposing litter. Further research should focus on quantifying the role of invertebrates in grass decomposition rates and nutrient cycling to improve knowledge of grassland ecology.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Litter invertebrates display greater differences among locations than grass species in a temperate grassland\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2024-12-03 09:48:00\",\"doi\":\"10.21203/rs.3.rs-5486939/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"decision\",\"content\":\"Revision requested\",\"date\":\"2025-01-06T19:25:49+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2024-12-23T17:53:16+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2024-12-19T20:25:20+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2024-12-17T20:22:39+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"191288029815857636899413263694505600859\",\"date\":\"2024-12-17T19:06:52+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"112125983909100412863181660009847710214\",\"date\":\"2024-12-17T03:56:43+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"215434777123412248503864391631989611988\",\"date\":\"2024-12-16T14:08:49+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"122383641250084091910658893122433448757\",\"date\":\"2024-12-16T13:54:05+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2024-12-15T17:00:51+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"51996499206285390435377339196839140553\",\"date\":\"2024-12-10T09:37:28+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2024-12-10T09:31:14+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2024-11-20T14:51:19+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"checksComplete\",\"content\":\"\",\"date\":\"2024-11-20T14:49:55+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Journal of Insect Conservation\",\"date\":\"2024-11-20T02:21:13+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"journal-of-insect-conservation\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"jico\",\"sideBox\":\"Learn more about [Journal of Insect Conservation](http://link.springer.com/journal/10841)\",\"snPcode\":\"10841\",\"submissionUrl\":\"https://submission.nature.com/new-submission/10841/3\",\"title\":\"Journal of Insect Conservation\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false}}],\"origin\":\"\",\"ownerIdentity\":\"970bf9b2-ba65-4d4e-9834-86ce0c9b26a4\",\"owner\":[],\"postedDate\":\"December 3rd, 2024\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"published-in-journal\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2025-04-14T16:15:20+00:00\",\"versionOfRecord\":{\"articleIdentity\":\"rs-5486939\",\"link\":\"https://doi.org/10.1007/s10841-025-00668-6\",\"journal\":{\"identity\":\"journal-of-insect-conservation\",\"isVorOnly\":false,\"title\":\"Journal of Insect Conservation\"},\"publishedOn\":\"2025-04-07 16:04:58\",\"publishedOnDateReadable\":\"April 7th, 2025\"},\"versionCreatedAt\":\"2024-12-03 09:48:00\",\"video\":\"\",\"vorDoi\":\"10.1007/s10841-025-00668-6\",\"vorDoiUrl\":\"https://doi.org/10.1007/s10841-025-00668-6\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-5486939\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-5486939\",\"identity\":\"rs-5486939\",\"version\":[\"v1\"]},\"buildId\":\"qtupq5eGEP_6zYnWcrvyt\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}