Predicting diversity in benthic macro-scale communities associated with mussel matrices in three Pacific ecoregions

Marine mussels are ubiquitous and their tough byssal threads allow for the formation of expansive, age-aggregated mats known as mussel matrices which can host marine invertebrates and algal macro-benthic communities playing an important role in food-web dynamics. Yet, despite the significant implications for biodiversity and intertidal ecosystem functioning, the role of mussel size, individual morphology and community arrangement in determining the structure of the associated community has never been investigated. Species representative of the green, brown and red marine algal phyla as well as polychaetes, crustaceans and gastropods were sampled from mussel matrices in the Guayaquil (GUAY), Humboldtian (HWS, HNS) and North American Pacific Fjordland (NAPF) marine ecoregions. Linear mixed effects modelling (LMM) and linear effects modelling (LEM) were used to determine the effect of mussel matrices as a predictor of species diversity by incorporating variability due to ecoregion and sampling site. Shell length and stratum index demonstrated significant effect on species richness (S obs ) and Menhinick's richness (D), while stratum index demonstrated an effect on species diversity (H). In the linear mixed model analysis, shell length explained most of the variation in Menhinick's richness (D) and observed species richness (S obs ), while stratum index explained most of the variation in (D) in the linear effects model. Our findings reveal that the level of complexity in mussel assemblages plays a major role in determining species diversity.

. On the contrary, some studies even seem to suggest that the faunal abundance associated with mussel beds is independent of mussel bed structure or species composition (Hammond & Griffiths, 2006).The majority of studies though suggest that mussel bed identity and characteristics do influence levels of epifaunal diversity (Cole, 2009;Kelaher, 2003).Specifically, mussel bed geometry is known to influence ecosystem function (Commito et al., 2014;Paquette et al., 2019), but, to date, few studies have focused specifically on the relationship between mussel bed geometry and species diversity and none have specifically dealt with the characteristics and relationship between mussel bed geometry and species diversity, which this study focuses on.Furthermore, developing an understanding of how the mussel bed structure influences diversity in different marine ecoregions is also of importance, especially when taking into account the demonstrated impact of latitude on epifaunal community characteristics (Sepúlveda et al., 2016).

| Mussel stratum index
Mussels are limited by space and food availability with the problem of space mitigated through mussel layering, also known as the mussel matrix.Guiñez and Castilla (1999) have derived a simple mathematical method for assigning a stratum index to mussel samples based on the height of the sampled mussel bed, the length of the shell, the area of the sampled matrix and the total area that would be covered by mussels if they formed a monolayer.Their model is derived from the research on predicting mussel matrix volume and area by Hosomi (1985).In this study, we model the relationship between species richness in the mussel communities under study using linear mixed effects modelling with the objective of identifying a biomarker of species richness for the ecoregions under study, where shell length, matrix depth and stratum index are the fixed effects and site and ecoregion are the random effects.Furthermore, we consider the practical use of matrix depth versus stratum index as a field application for differentiating matrix layers and for determining species richness.
A comprehensive understanding of these benthic macro-scale communities as they relate to mussel assemblages may provide a framework for coastal management (Bateman & Bishop, 2017;Engle, 2008).This study was conducted with the primary aim of understanding the role of mussel size and mussel matrix structure on benthic macro-scale community diversity and richness in different marine ecoregions.

| Study site and ecoregions
The current study borrows from the Census of Marine Life's concept of mapped classifications of marine areas (Marine Ecoregions of the World (MEOW) -Census of Marine Life Maps and Visualization 2010) and put forward by Spalding et al. (2007) for the purpose of representing ecologically important coastal areas for comparisons on multiple geographic scales.Spalding et al. (2007) comprised a list of definitions of the classification system that is parallel to the classification established by the International Union for the Conservation of Nature (Fosberg, 1976), which ranges from largest in scale to smallest consisting of Realms, Provinces and Ecoregions.All of the ecoregions included in this study contain mussel species that are broadly endemic to each Province.The ecoregions under consideration include the North American Fjordland (NAPF) between ~50° and 59° N, the Guayaquil (GUAY) ecoregion between ~0° and 6° S and the Humboldtian ecoregion between ~12° and 26° S (Figure 1).
In order to increase the spatial resolution of the Humboldtian ecoregion, two divisions are proposed based on the width of the continental margin: the Humboldtian wide-shelf (HWS) ecoregion between 12° S and 13° S is approximately 225 km from the edge of the continental shelf to the coastline; and the Humboldtian narrow-shelf (HNS) ecoregion between 15° S and 16° S, and 23° S and 24° S, with a continental shelf approximately half that of the HWS (Krabbenhöft et al., 2004;Strub, 1998).Four sites within the NAPF ecoregion, three sites within the HWS ecoregion and five sites within the HNS ecoregion were chosen based on the presence of endemic mussel species (Figure 1).The focus on areas with endemic mussel species was to ensure that the epifaunal communities and diversity patterns will better reflect the natural state of these ecoregions.Geographic coordinates for the NAPF, GUAY, HWS, and HNS ecoregions and sites within those ecoregions are shown in Table 1.
Mussel plots were selected at each site where mussels were present in the high-to-low intertidal zone.A minimum of 5 replicate plots with a minimum of 20% coverage of a 70 × 50 cm quadrat were chosen either along a transect vertical to the waterline or where mussels formed narrow but continuous bands.Where it was possible to sample the same site, care was taken to not re-sample the same quadrats by marking the location of each sampled assemblage using a reel transect.The total number of replicate quadrats (n) by ecoregion is shown in Table 2. Mussel plots were sampled in the GUAY ecoregion, in the HWS ecoregion and at Reserva Punta San Juan (PSJ) in the HNS ecoregion in the austral summer of 2017, mussel plots were sampled at one new site in the HNS ecoregion in 2018 and additional mussel plots were sampled at the PSJ site in the HNS ecoregion in 2019.Mussel plots at sites within the NAPF ecoregion were sampled separately in the boreal summers of 2017 and 2018.No re-sampling of any of the plots occurred during any of the sampling events.

| Mussel species and macrofaunal sampling
The two most commonly found species of mussels (family considered to be "bioengineer" species (Jones et al., 1997;Prado & Castilla, 2006).In Peru, P. purpuratus is found in the mid-to-low intertidal zone while S. algosus is typically found overlapping with P. purpuratus in the mid zone, becoming abundant and the predominant mussel species in the low zone, with some spatial overlap between the two species (Tokeshi & Romero, 2000).Assemblages of P. purpuratus and S. algosus were the targets for measuring abundances of invertebrates and algae that exist as microcosms within the mussel assemblages at Playa Ensenada, Playa Farallones and Playa Palmeras respectively.
In the NAPF ecoregion, two species of mussels found in the intertidal zone are Mytilus trossulus and Mytilus californianus, with the introduced species Mytilus galloprovincialis and Mytilus edulis reported for the Pacific Northwest.M. trossulus is typically found in concentrated assemblages well above the zero-foot tidal height assigned to the mean low water soundings, whereas M. californianus typically occurs in stands of one to few individuals well below the zone where assemblages of M. trossulus are found (L.Wilbur, unpublished data).As M. edulis and M. galloprovincialis occur in intertidal areas in Sitka Sound alongside stands of M. trossulus, identification between the species using shell morphology technique is not considered reliable (Hilbish et al., 2002); therefore, a consideration of the M. edulis/trossulus/galloprovincialis complex has been suggested when designating species identification in the field in the absence of genetic analysis and mussels from the NAPF ecoregion will be referred henceforth as MYTCOM.
Intertidal assemblages associated with mussel communities were sampled by counting invertebrates and the thalli of algae attached to the outer shells of mussels at each site with a 70 × 50 cm quadrat strung to provide 10 transects with 100 intersections (Engle, 2008).
In order to include algal sporelings and invertebrates that might be missed living below the top layers of mussels, we sub-sampled the quadrat by selecting 10 intersections across the quadrat with a random numbers calculator.First, a plastic Vernier calliper with a retractable stainless-steel rod was inserted into the mussel assemblage to measure the depth of the matrix to the nearest 0.5 mm from the tip of the highest mussel to where the rod touched the rock substrate (Prado & Castilla, 2006).Similar to the methods used by Guiñez and Castilla (1999), mussels were then removed from under each intersection using a 20 × 60 mm stainless steel spatula to fill a 100 mm 2 frame.
The mussel matrices were then placed in a tray and covered with a shallow layer of seawater for approximately 30 min to allow mobile animals to emerge from the shells.Invertebrates and algal sporelings ≥1 mm in length living inside and on the mussels were examined using a hand lens and Nikon field 20x stereoscope and counted.Reference samples were preserved in 70% EtOH (invertebrates) and seawater TA B L E 1 Geographic coordinates for all ecoregions, subecoregions within ecoregions, and sites within ecoregions and subecoregions where mussel assemblages were sampled for this study.TA B L E 2 Units of central tendency for (a) fixed effects mean ± SD and range and (b) response variables (median denoted by *) or mean ± SD calculated from mussel plot data pooled for each ecoregion.GUAY (n = 11 5.9 (± 0.10) Abbreviations: NAPF, North American Pacific Fjordland; GUAY, Guayaquil; HWS, Humboldtian wide-shelf; HNS, Humboldtian narrow-shelf.(algae) for transport back to a field-based laboratory, where they were identified to the lowest taxonomic classification possible using identification keys and the available regional reference materials (Dawson et al., 1964;Howe, 1914;Kozloff & Price, 1987;Romero, 2002).
Additional information on the identification of polydorid species including a possible invasive polychaete can be found in Appendix A.

| Calculating the stratum index of mussel layers
The mussel matrices that were sampled from the intersections of the transects were used for calculating the stratum index of each respective plot.Mussels were measured lengthwise to the nearest 0.5 mm using a Vernier calliper starting with the mussels that served as the centre of attachment followed by mussels attached to the central mussel >1.5 mm in length (dead mussel shells and empty shells were discarded) for a total of n = 15 per plot.Mussels <1.5 mm were considered to recruit and can cause a skew in the population data (Guiñez & Castilla, 1999).For each mussel plot, the appropriate unit of central tendency was calculated based on the distribution of the variables shell length (L) and matrix depth (M).The total area under the matrix (total occupation area (S), Hosomi (1985) for shell lengths equal to or greater than 10 mm in length was calculated according to the formula: and for shell lengths less than 10 mm in length according to the formula: The results from the calculations from each of the raw shell lengths were summed using the formula: where phi is a ratio used to measure body proportions in plants and animals, and phi = 1.6180339887495 (Hosomi, 1985).The summed values were used to represent the variable stratum index (SI) for each corresponding plot (Guiñez & Castilla, 1999;Hosomi, 1987).The distribution of the values for shell length (L), matrix depth (M), stratum index (SI) and species richness values calculated for all plots in this study were evaluated for normal distributions with the Anderson-Darling test (p > .05).We calculated mean for observed species richness (S obs ) and Shannon-Wiener diversity (H) using randomized re-sampling with bootstrapped standard deviations (SD) from algorithms provided in EstimateS software version 9.1 (Gotelli & Colwell, 2011).

| Statistical analysis
We sought to use several measures of diversity that would account for the number of species, abundance and evenness that would allow us to explore multiple dependent variables in our analysis.We used Menhinick's (D) index as one of the species diversity measures because this method accounts for abundance in a sample and effectively compares samples of different sizes (Menhinick, 1964).The formula is D = S /√N, where N = the number of individuals sampled and S is the observed species richness; and Shannon-Wiener diversity index ∑ pi ln(pi) (Shannon, 1948), where pi = the relative abundance of individuals for each species.Because shell length was used to calculate the stratum index, we standardized the independent variables in order to adjust for collinearity (Allen, 1997) using the formula: Frequency distributions for all of the variables were analysed for normality using the Anderson-Darling test.
Linear mixed effects and linear effects modelling were performed on the mussel plot data with stratum index, matrix depth, and shell length as the fixed effects, Mehinick's richness (D), and Shannon-Wiener diversity (H') as the response variables, and ecoregion and/or site as the random effects.Akaike information criteria (AIC) scores were calculated for each model to determine the random effects that were the best fit; lower AIC scores indicated the better model (Akaike, 1974).The linear mixed effect model (LMEM) we used is a random-slope/random-intercept model, which measures the variation in the fixed effects versus the response variables for random effects (Bates et al., 2015) using the following formula: To provide a visualization of fixed effects on diversity, independent variables from LMEM models that resulted in statistically significant results (p < .05)were then modelled using linear effects (LEM) with the random intercept/fixed slope formula: Imer (dependent variable~independent variable + (1|random effect 1) + (1|random effect 2)) and plotted using the ggPlot package to show a random intercept and fixed slope plot (Wickham, 2011), for comparison among ecoregions.All linear effects modelling was performed using the lmerTest (Kuznetsova et al., 2017) in R 4.0.2(R Core Team, 2021) using RStudio v.1.3.1093(RStudio Team, 2009).

| Macro-scale patterns of intertidal benthic species
A total of 72 species representing 12 phyla were found in mussel plots in the NAPF ecoregion; 9 species from 5 phyla were found within the GUAY ecoregion, 49 species from 12 phyla were found  et al., 2011).A table of species encountered in the mussel plots by ecoregion and site can be found in Appendix B.

| Mussel shell length and stratum index as predictors of diversity in mussel aggregations
We tested the appropriateness of mussel matrices' physical characteristics in predicting species diversity.A total of 89 mussel plots were sampled, with a total of 89 mussel matrix replicates sampled for the biodiversity analysis, and 1118 mussels were measured lengthwise to calculate the stratum indices.Due to omissions in the sampling of shell lengths in 3 of the 89 plots, 86 plots were included in the analysis.Summary statistics were calculated for mean values of the fixed effects that had a normal distribution, and the range of values from minimum to maximum for stratum index, matrix depth and shell length, along with sample size (number of mussel plots sampled) for each respective ecoregion (Table 2).
The lowest AIC scores from the random slope/random intercept LMEM models included ecoregion as the random effect and site as the nested random effect.The random slope/random intercept LMEM produced significant effects of varying magnitude for the standardized shell length and stratum index across all three diversity measures used, with shell length accounting for most of the variance (Table 3).Satterthwaite's method for approximating degrees of freedom is used by the lmer command to produce p-values for the t-test with significance at alpha adjusted to ≤0.02 to correct for multiple comparisons that showed significant differences in the S obs model for shell length (estimate = 4.91, p = .004),in the D model for shell length (estimate = 0.39, p = .02)and the H model for shell length (estimate = 0.18, p = .0).The conditional r-squared values indicated that the strength of the relationship of the random and fixed effects for all models was strong; however, when the raw values for shell length and stratum index were analysed individually in the random intercept/fixed slope LEM model, only shell length was significant for variation in both richness S obs (Est.= 0.20, p = .02)and richness D (Est.= 0.03, p = .001)(Table 4).The slopes for the LEM suggest that S obs increases with increasing shell length, with ecoregion explaining much of the variation.The slopes also suggest that Menhinick's (D) increases with increasing shell length with an increase in the stratum index (with intercepts varying across ecoregions).Although the AIC score for the model of shell length v D is the lowest for both models, the model for stratum index explained most of the variation in D.
The results from the LEM suggest that species richness can be predicted from shell length and stratum index with regional variation (Figure 2).

| DISCUSS ION
Shell size, matrix depth and the complexity of mussel aggregations are linked to age class and various environmental factors that exist where the mussel aggregations are found (Alvarado & Castilla, 1996).
Here, we examined the physical geometry of the mussel matrices and their associated benthic macro-scale communities across a broad TA B L E 3 Estimates of variances and standard errors with conditional r-squared from the random slope/random intercept linear mixed models to analyse the effects of matrix depth, shell length and stratum index on species richness (S), Menhinick's richness (D) and Shannon-Wiener (H') diversity in mussel plots sampled from the GUAY, HWS, HNS and NAPF ecoregions.Note: In these models, the slopes of the dependent variables versus the independent variables are the same among all ecoregions, while the intercepts vary.Of the random effects, ecoregion explained more of the variation than did site for both models  approaches (Aronson & Precht, 1995), fractal analysis (Commito & Rusignuolo, 2000) or three-dimensional surface rugosity (Parravicini et al., 2006), we suggest that mussel shell length (an individual-based measure) is potentially the most important component of mussel bed complexity and can be used as a biomarker for predicting species richness in mussel matrices across different marine ecoregions.That perhaps is not surprising as shell length is an important component of SI, but also perhaps shell length is a strong indicator of shell surface area available to epifaunal organisms.Larger mussels are generally older mussels, which suggests a longer period of time for colonization by other species.Furthermore, larger (thus potentially older) mussels are the main locus of attachment for younger, smaller mussels (Commito et al., 2014).As new mussels recruit onto the existing mussels, this leads to an increase in surface area for epifaunal organisms to colonize on, as well as increasing the spatial complexity, which may lead to a further increase in species richness.While species richness (S obs ) may have a stronger positive correlation with shell length, species richness (D) may be predicted with a higher level of precision based on model parameters.
Our study has demonstrated that species richness is higher in multi-layered matrices regardless of whether the matrix is defined by the depth or by the more complicated stratum index formula, suggesting that by providing more habitable substrate, the surface area of the matrix may be a function of species richness.This result is confirmed by other studies that suggest that mussel bed complexity is a more important aspect in determining levels of diversity compared to the availability of nutrients, indicating perhaps a more top-down effect of community composition than a bottom-up one (Firstater et al., 2011).Along the same lines, it has been shown that invasive mussel species that contribute to an increase in mussel bed complexity contribute to an increase in diversity, more so than native mussel species (Gestoso et al., 2013).Here, we show the importance of stratum index as a predictor of community heterogeneity within mussel matrices.Prado and Castilla (2006) recognized the relevance of a stratum index for defining environmental complexity in mussel matrices and utilized matrix depth as an appropriate field method for quantifying mono-and multi-layered matrices.We have shown that measuring matrix depth in the field was indeed found to be an efficient method for describing matrix layers, nevertheless, calculating the stratum index is useful due to the incorporation of area and shell length, both of which assist in determining the level of structural complexity and predominant age class of the mussels.
Here, we have successfully recruited these easily observational factors (matrix depth and shell length) as well as the formula for stratum index developed by Hosomi (1985) to demonstrate its usefulness for predicting species richness in complex mussel aggregations in the four ecoregions under study.
Mytelidae) in the Humboldtian ecoregion are Perumytilus purpuratus (Lamarck 1819) and Semimytilus algosus (Gould 1850), with Brachidontes spp. as the dominant genus north of 5 degrees latitude in the GUAY ecoregion (L.Wilbur unpubl.data).P. purpuratus and S. algosus are associated with high biodiversity and are thus 14390485, 2023, 2, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/maec.12741 by University Of Aberdeen, Wiley Online Library on [24/04/2023].See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions)on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License F I G U R E 1 Overview map of the western portion of the northern and southern hemispheres in the eastern Pacific and associated ecoregions (insets) and the study sites (black dots) where mussel plots were sampled.
14390485, 2023, 2, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/maec.12741 by University Of Aberdeen, Wiley Online Library on [24/04/2023].See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions)on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License 14390485, 2023, 2, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/maec.12741 by University Of Aberdeen, Wiley Online Library on [24/04/2023].See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions)on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License within the HWS ecoregion and 101 species from 11 phyla were found within the HNS ecoregion.Filamentous and foliose green algae were predominantly found in the transects in the GUAY and HNS ecoregions, while corticated red algae were the most predominant in the HWS ecoregion and leathery brown macrophytes were the most predominant in the NAPF ecoregion.Other than mytilids, molluscs were dominant in the GUAY, HWS and HNS ecoregions, followed by arthropods and annelids.Of the molluscs, the limpet Scurria viridula was the most common species in the HWS and HNS ecoregions while the periwinkle Echinolittorina paytensis was most commonly found in the GUAY ecoregion.Of the arthropods, the barnacle Jehlius cirratus was the most commonly found, and among polychaete annelids, Perinereis was commonly found in the HWS and HNS ecoregions, followed by a spionid polychaete (Boccardia cf.wellingtonensis) at Reserva Punta San Juan in the HNS ecoregion.In the NAPF ecoregion, arthropods dominated the mussel plots with Balanus glandula as the most commonly found species, followed by Semibalanus cariosus and Amphibalanus sp., a genus of barnacle known to be invasive in the eastern north Pacific(Carlton ecoregional scale in an effort to better understand the characteristics of mussel matrices, particularly when attempting to predict patterns in diversity.It has been suggested that environmental factors often play a role in the abundance and presence of certain species, and because the marine ecoregions are partly defined by local conditions(Ibanez- Erquiaga et al., 2018; Valqui et al., 2021), "ecoregions" as random effects were incorporated into the modelling analysis.Therefore, our results support small-scale experimental results which show that mussel bed complexity is an important factor in determining species composition and abundance(Koivisto & Westerbom, 2010).Although over time, studies have utilized increasingly complex approaches in an effort to characterize mussel bed structure, for example, chain-length TA B L E 4 Variances and significant values from the fixed slope/random intercept linear effects (LEM) models of the respective relationships between the fixed effect shell length versus species richness (S) and Menhinick's (D) and stratum index versus (S) and (D), calculated from mussel plots from all ecoregions in this study.
(S) and (D) (ss = 3.02 and sD = 0.23).A comparison of the models that demonstrated significance at p < .05(in bold) (shell length vs. D and stratum index vs.D) from the lmer test was performed with a post hoc Chi-squared ANOVA.The two models were significantly different (p < .001);corresponding AIC scores are given in the right-hand column of the table.

F I G U R E 2
Line graphs of the fixed slope/random intercept model from the linear effects modelling (LEM) fixed (shell length and stratum index) and random (ecoregion and site) effects versus species richness for mussel plots sampled from all ecoregions.a) Shell length and observed species richness (S obs ); b) shell length and Menhinick's richness (D); and c) stratum index and Menhinick's richness (D).Models were chosen from an alpha <0.05 with the conditional r-squared (fixed and random effects) value provided in the bottom right corner of each graph.Each ecoregion is represented by the coloured line indicated in the legend.Abbreviations for each ecoregion are as follows: GUAY = Guayaquil, HNS = Humboldtian Narrow-shelf, HWS = Humboldtian Wide-Shelf and NAPF = North American Pacific Fjordland.
of invertebrates and algae encountered in intertidal mussel communities at sites within the Guayaquil (GUAY), Humboldtian Wide-Shelf (HWS) and Humboldtian Narrow-Shelf (HNS) ecoregions in this study.Site abbreviations are given where particular species were found.

Table B .2. Table of invertebrates and algae encountered in intertidal mussel communities at sites within the NAPF ecoregion. Site abbreviations are given where particular species were found. North American Pacific Fjordland Chlorophyta
Stereoscopic images of the specimen proposed as the polychaete Boccardia wellingtonensis (family Spionidae, Read, 1975) collected from Reserva Punta San Juan in the Marcona district in the Humboldtian ecoregion of Peru.Counter-clockwise from top left, (a)-(b) dorsal view of prostomium, showing the prostomial incision characteristic of B. wellingtonensis, (c) bristles visible at the top of spines on setiger 5, and (d) notopodial spines present on last segments.Site abbreviations are as follows: ACA, Playa Acapulco; ENU, El Nuro; PEN, Playa Ensenada; PFA, Playa Farallones; PPA, Playa Palmeras; PSJ, Punta San Juan (with sites N5n, N5s, S4 and S5) UOA, Universidad de Antofagasta., 2023, 2, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/maec.12741 by University Of Aberdeen, Wiley Online Library on [24/04/2023].See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions)on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/maec.12741 by University Of Aberdeen, Wiley Online Library on [24/04/2023].See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions)on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License F I G U R E A . 1 14390485, 2023, 2, Downloaded from https://onlinelibrary.wiley.com/doi/10.1111/maec.12741 by University Of Aberdeen, Wiley Online Library on [24/04/2023].See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions)onWiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License | 13 of 15 WILBUR et al.14390485