Species richness in agroecosystems: the effect of landscape, habitat and farm management

It has been suggested that biodiversity in agroecosystems depends on both landscape heterogeneity and farm management, but at the same time, studies of biodiversity in relation to both landscape variables and farm management are rare. We investigated the species richness of plants, butterflies, carabids, rove beetles and the diversity of spiders in cereal fields, leys (grass and clover crop) and semi-natural pastures at 16 farms in Central East Sweden. The farms were divided into eight pairs of one conventional and one organic farm to enable us to separate the effects of landscape and farm management on biodiversity. The pairing was based on land use, location, and landscape features. Species richness of different taxonomic groups was generally not correlated. There were no differences in species richness between the farming systems, except for carabids that had higher numbers of species on conventional farms. The species richness generally increased with landscape heterogeneity on a farm scale. Habitat type had a major effect on the species richness for most groups, with most species found in pastures and leys. The correlations between species richness and landscape variables on a farm scale, and not on a scale of multiple farms, identify farmers as the important decision-maker in conservation issues for these taxonomic groups. We discuss the role of species richness of pests' natural enemies for biological control and conservation strategies of the more common species in the agricultural landscape.     

Steppe-like grassland as a refuge of the wild edible and medicinal plant species in anthropogenic landscape in northeastern Croatia

Habitat fragmentation and modification have been recognized as the most serious threats to biodiversity. In Croatia, the intensive human activities resulted in the landscape transformation, reduction of grassland areas and loss of species number. Increasingly threatened are the wild edible and medicinal plants which contribute remarkably to the biodiversity of different grassland types. In northeastern Croatia, one of the last remnants of the steppe-like grassland remained preserved in a very complex anthropogenic landscape. We examined the diversity of vascular plants occurring on this small habitat patch. Among the relatively high number of plant taxa (in a total of 177) found during the two years of investigations, most could be used both for human nutrition (28 taxa), and for the treatment of a wide range of diseases (60 taxa), while some taxa have nutritional value for livestock (17 taxa). All 23 recorded taxa with edible and medicinal properties are threatened in Croatia, 21 are threatened across Europe, and two are Red Listed taxa. Our results showed that small and isolated steppe-like grassland represents an important refuge for variety of valuable plant species in intensively used landscapes. Accordingly, such small habitats should be, at least locally, effectively protected.     

Organic farming improves pollination success in strawberries

Pollination of insect pollinated crops has been found to be correlated to pollinator abundance and diversity. Since organic farming has the potential to mitigate negative effects of agricultural intensification on biodiversity, it may also benefit crop pollination, but direct evidence of this is scant. We evaluated the effect of organic farming on pollination of strawberry plants focusing on (1) if pollination success was higher on organic farms compared to conventional farms, and (2) if there was a time lag from conversion to organic farming until an effect was manifested. We found that pollination success and the proportion of fully pollinated berries were higher on organic compared to conventional farms and this difference was already evident 2-4 years after conversion to organic farming. Our results suggest that conversion to organic farming may rapidly increase pollination success and hence benefit the ecosystem service of crop pollination regarding both yield quantity and quality.     

A global synthesis of the effects of diversified farming systems on arthropod diversity within fields and across agricultural landscapes

Agricultural intensification is a leading cause of global biodiversity loss, which can reduce the provisioning of ecosystem services in managed ecosystems. Organic farming and plant diversification are farm management schemes that may mitigate potential ecological harm by increasing species richness and boosting related ecosystem services to agroecosystems. What remains unclear is the extent to which farm management schemes affect biodiversity components other than species richness, and whether impacts differ across spatial scales and landscape contexts. Using a global metadataset, we quantified the effects of organic farming and plant diversification on abundance, local diversity (communities within fields), and regional diversity (communities across fields) of arthropod pollinators, predators, herbivores, and detritivores. Both organic farming and higher in‐field plant diversity enhanced arthropod abundance, particularly for rare taxa. This resulted in increased richness but decreased evenness. While these responses were stronger at local relative to regional scales, richness and abundance increased at both scales, and richness on farms embedded in complex relative to simple landscapes. Overall, both organic farming and in‐field plant diversification exerted the strongest effects on pollinators and predators, suggesting these management schemes can facilitate ecosystem service providers without augmenting herbivore (pest) populations. Our results suggest that organic farming and plant diversification promote diverse arthropod metacommunities that may provide temporal and spatial stability of ecosystem service provisioning. Conserving diverse plant and arthropod communities in farming systems therefore requires sustainable practices that operate both within fields and across landscapes.     

Do differences in food web structure between organic and conventional farms affect the ecosystem service of pest control?

While many studies have demonstrated that organic farms support greater levels of biodiversity, it is not known whether this translates into better provision of ecosystem services. Here we use a food-web approach to analyse the community structure and function at the whole-farm scale. Quantitative food webs from 10 replicate pairs of organic and conventional farms showed that organic farms have significantly more species at three trophic levels (plant, herbivore and parasitoid) and significantly different network structure. Herbivores on organic farms were attacked by more parasitoid species on organic farms than on conventional farms. However, differences in network structure did not translate into differences in robustness to simulated species loss and we found no difference in percentage parasitism (natural pest control) across a variety of host species. Furthermore, a manipulative field experiment demonstrated that the higher species richness of parasitoids on the organic farms did not increase mortality of a novel herbivore used to bioassay ecosystem service. The explanation for these differences is likely to include inherent differences in management strategies and landscape structure between the two farming systems.     

Benefits of habitat restoration to small mammal diversity and abundance in a pastoral agricultural landscape in mid-Wales

Changes in agricultural practice are predicted across the UK following agricultural reform driven by government policy. The suitability of agri-environment schemes for many species is currently debated because limited quantitative data are collected. In order to understand the changes to biodiversity due to agri-environment schemes, there is a need for studies to not just compare biodiversity and species composition in and out of agri-environment areas, but to factor in the influence of temporal habitat changes. In this study, we investigate the suitability of an agri-environment initiative to support and enhance a small mammal fauna among pastoral hill farms in mid-Wales. Grazed and ungrazed woodlands, riparian habitats, and broadleaf plantations, were compared for small mammal abundance and diversity following a trapping study. Mammal diversity was similar across habitats, though abundance varied significantly. A principle component analysis identified that mammal abundance clustered into three main habitat groups separated by seral stage (early, mid, late). No relationship between mammal abundance and stock grazing was found. A canonical correspondence analysis confirmed that vegetation structure was important in explaining the distribution of captures of mammal species across the landscape. The results for habitat type, and habitat context, suggest that a mix of vegetation seral stages, reflecting a varied vegetation structure, is important to maintain small mammal diversity and abundance across the study area. Heterogeneity in structural diversity at the landscape scale is important to maintain a variety of ground-dwelling mammal species, and particularly because trends in countryside surveys show that woodlands are skewed towards late seral stages. Habitat heterogeneity can be maintained because the hill farms neighbour each other, and the farmers co-operate as a group to manage the landscape. Habitat diversity is therefore possible. These results help us to advocate, and anticipate, the benefits of groups of farms within a landscape.     

Impacts of organic and conventional dairy farmer attitude,behaviour and knowledge on farm biodiversity in Ireland

Agricultural intensification has caused significant declines in biodiversity. Agri-environmental schemes (AES), including organic farming, are thought to benefit biodiversity. However, under similar production conditions and in comparable locations and schemes, farms are not managed in the same way, with variable consequences for production and the environment. Understanding farmer attitude, behaviour and knowledge of the environment and their impacts on biodiversity may help us understand the variable impacts of AES on biodiversity. We combined a sociological survey of nine organic and eight conventional dairy farmers (measuring environmental and achievement attitudes; environmental and production-orientated behaviours and environmental knowledge), in Ireland, with a biodiversity (using plant richness as an indicator) assessment of their farms. We found higher plant richness on organic farms than conventional. Organic and conventional farmers had similar attitudes to farming achievement and the environment but organic farmers were better informed about environmental issues and carried out more environmentally orientated behaviours. Biodiversity was positively related to the amount of environmentally orientated behaviours and negatively related to production-orientated behaviours carried out on farms. Organic farmers who had more positive attitudes to the environment and were better informed about it had higher biodiversity on their farms compared to less positively inclined, less informed organic farmers. We show disparities between attitude and actual behaviour in relation to the environment, with organic farmers sharing similar attitudes to conventional farmers, but more prepared to inform themselves about and carry out environmentally friendly farming. Results indicate that organic farming and environmentally orientated behaviours benefit biodiversity and that there is an important link between farmer environmental attitudes and knowledge and the beneficial effects of organic farming on biodiversity. We encourage conservation orientated thinking and better environmental education among farmers, including those who already participate in an AES. This way, the benefits of the AES for the environment may be maximised.     

Organic Farming: Biodiversity Impacts Can Depend on Dispersal Characteristics and Landscape Context

Organic farming, a low intensity system, may offer benefits for a range of taxa, but what affects the extent of those benefits is imperfectly understood. We explored the effects of organic farming and landscape on the activity density and species density of spiders and carabid beetles, using a large sample of paired organic and conventional farms in the UK. Spider activity density and species density were influenced by both farming system and surrounding landscape. Hunting spiders, which tend to have lower dispersal capabilities, had higher activity density, and more species were captured, on organic compared to conventional farms. There was also evidence for an interaction, as the farming system effect was particularly marked in the cropped area before harvest and was more pronounced in complex landscapes (those with little arable land). There was no evidence for any effect of farming system or landscape on web-building spiders (which include the linyphiids, many of which have high dispersal capabilities). For carabid beetles, the farming system effects were inconsistent. Before harvest, higher activity densities were observed in the crops on organic farms compared with conventional farms. After harvest, no difference was detected in the cropped area, but more carabids were captured on conventional compared to organic boundaries. Carabids were more species-dense in complex landscapes, and farming system did not affect this. There was little evidence that non-cropped habitat differences explained the farming system effects for either spiders or carabid beetles. For spiders, the farming system effects in the cropped area were probably largely attributable to differences in crop management; reduced inputs of pesticides (herbicides and insecticides) and fertilisers are possible influences, and there was some evidence for an effect of non-crop plant species richness on hunting spider activity density. The benefits of organic farming may be greatest for taxa with lower dispersal abilities generally. The evidence for interactions among landscape and farming system in their effects on spiders highlights the importance of developing strategies for managing farmland at the landscape-scale for most effective conservation of biodiversity.     

Parasitoid diversity reduces the variability in pest control services across time on farms

Recent declines in biodiversity have increased interest in the link between biodiversity and the provision and sustainability of ecosystem services across space and time. We mapped the complex network of interactions between herbivores and parasitoids to examine the relationship between parasitoid species richness, functional group diversity and the provision of natural pest control services. Quantitative food webs were constructed for 10 organic and 10 conventional farms. Parasitoid species richness varied from 26 to 58 species and we found a significant positive relationship between parasitoid species richness and temporal stability in parasitism rates. Higher species richness was associated with lower variation in parasitism rate. A functional group analysis showed significantly greater parasitoid species complementarity on organic farms, with on average more species in each functional group. We simulated parasitoid removal to predict whether organic farms experienced greater robustness of parasitism in the face of local extinctions. This analysis showed no consistent differences between the organic and conventional farm pairs in terms of loss of pest control service. Finally, it was found that the different habitats that make up each farm do not contribute equally to parasitoid species diversity, and that hedgerows produced more parasitoid species, significantly more so on organic farms.     

Species Richness and Assemblages in Landscapes of Different Farming Intensity – Time to Revise Conservation Strategies?

Worldwide conservation goals to protect biodiversity emphasize the need to rethink which objectives are most suitable for different landscapes. Comparing two different Swedish farming landscapes, we used survey data on birds and vascular plants to test whether landscapes with large, intensively managed farms had lower richness and diversity of the two taxa than landscapes with less intensively managed small farms, and if they differed in species composition. Landscapes with large intensively managed farms did not have lower richness than smaller low intensively managed farms. The landscape types were also similar in that they had few red listed species, normally targeted in conservation. Differences in species composition demonstrate that by having both types of agricultural landscapes regional diversity is increased, which is seldom captured in the objectives for agro-environmental policies. Thus we argue that focus on species richness or red listed species would miss the actual diversity found in the two landscape types. Biodiversity conservation, especially in production landscapes, would therefore benefit from a hierarchy of local to regional objectives with explicit targets in terms of which aspects of biodiversity to focus on.     

Sustained functional composition of pollinators in restored pastures despite slow functional restoration of plants

Habitat restoration is a key measure to counteract negative impacts on biodiversity from habitat loss and fragmentation. To assess success in restoring not only biodiversity, but also functionality of communities, we should take into account the re‐assembly of species trait composition across taxa. Attaining such functional restoration would depend on the landscape context, vegetation structure, and time since restoration. We assessed how trait composition of plant and pollinator (bee and hoverfly) communities differ between abandoned, restored (formerly abandoned) or continuously grazed (intact) semi‐natural pastures. In restored pastures, we also explored trait composition in relation to landscape context, vegetation structure, and pasture management history. Abandoned pastures differed from intact and restored pastures in trait composition of plant communities, and as expected, had lower abundances of species with traits associated with grazing adaptations. Further, plant trait composition in restored pastures became increasingly similar to that in intact pastures with increasing time since restoration. On the contrary, the trait composition of pollinator communities in both abandoned and restored pastures remained similar to intact pastures. The trait composition for both bees and hoverflies was influenced by flower abundance and, for bees, by connectivity to other intact grasslands in the landscape. The divergent responses across organism groups appeared to be mainly related to the limited dispersal ability and long individual life span in plants, the high mobility of pollinators, and the dependency of semi‐natural habitat for bees. Our results, encompassing restoration effects on trait composition for multiple taxa along a gradient in both time (time since restoration) and space (connectivity), reveal how interacting communities of plants and pollinators are shaped by different trait–environmental relationships. Complete functional restoration of pastures needs for more detailed assessments of both plants dispersal in time and of resources available within pollinator dispersal range.     

Differential responses of plants,reptiles and birds to grazing management,fertilizer and tree clearing

The relative effects of tree clearing, increased livestock densities and nutrient enrichment have rarely been compared across markedly different organism types, but negative effects are generally predicted. In contrast, adoption of rotational grazing is thought to benefit biodiversity in pastures but there are few supporting data. We examined the response of native plants, birds and reptiles to livestock management in south‐eastern Australia. We selected 12 pairs of rotationally and continuously grazed farms. Two 1‐ha plots were established in native pastures on each farm, one cleared and the second still retaining woodland tree cover. Stocking rates, fertilizer histories and landscape tree cover varied among farms. The abundance and richness of all taxa was lower in cleared pastures. The less mobile organisms (reptiles and plants) were positively correlated with tree cover at landscape scales, but only when trees were present at the plot scale. This pattern was driven by a few observations in landscapes with approximately 50% tree canopy cover. Neither bird abundance nor richness was correlated with stocking rates or nutrient enrichment, but plant richness responded negatively to both. The response of reptiles varied, declining with nutrient enrichment but positively correlated with livestock densities. These responses may be partly interpreted within the context of prior filtering of species pools through long‐term grazing pressure. No taxa responded positively to rotational grazing management. We predict that reductions in livestock density and soil nutrients will directly benefit plants and less so reptiles, but not birds. Indirect benefits are predicted for birds and reptiles if management increases persistence of trees within paddocks. Although some forms of rotational grazing can increase woodland tree recruitment, rotational grazing in itself is unlikely to enhance diversity.     

The effect of fragment area on site‐level biodiversity

Habitat fragmentation accompanies habitat loss, and drives additional biodiversity change; but few global biodiversity models explicitly analyse the effects of both fragmentation and loss. Here we propose and test the hypothesis that, as fragment area increases, species density (the number of species in a standardised plot) will scale with an exponent given by the difference between the exponents of the species–area relationships for islands (z ~ 0.25) and in contiguous habitat (z ~ 0.15), and test whether scaling varies between land uses. We also investigate the scaling of overall abundance and rarefaction‐based richness, as some mechanisms make different predictions about how fragment area should affect them. The relevant data from the taxonomically and geographically broad PREDICTS database were used to model the three diversity measures, testing their scaling with fragment area and whether the scaling exponent varied among land uses (primary forest, secondary forest, plantation forest, cropland and pasture). In addition, the consistency of the response of species density to fragment area was tested across three well represented taxa (Magnoliopsida, Hymenoptera and ‘herptiles’). Species density and total abundance showed area‐scaling exponents of 0.07 and 0.16, respectively, and these exponents did not vary significantly among land uses; rarefaction‐based richness by contrast did not increase consistently with area. These results suggest that the area‐scaling of species density is driven by the area‐scaling of total abundance, with additive edge effects (species moving into the small fragments from the surroundings) opposing – but not fully overcoming – the effect of fragment area on overall density of individuals. The interaction between fragment area and higher taxon (plants, vertebrates and invertebrates), which remained in the rarefied richness model, indicates that mechanisms may vary among groups.     

The living heart: Climate gradients predict desert mountain endemism

Mountain regions are centers of biodiversity endemism at a global scale but the role of arid‐zone mountain ranges in shaping biodiversity patterns is poorly understood. Focusing on three guilds of taxa from a desert upland refugium in Australia, we sought to determine: (a) the relative extent to which climate, terrain or geological substrate predict endemism, and (b) whether patterns of endemism are complimentary across broad taxonomic guilds. We mapped regional endemism for plants, land snails, and vertebrates using combined Species Distribution Models (SDMs) for all endemic taxa (n = 82). We then modelled predictors of endemism using Generalised Additive Models (GAMs) and geology, terrain, and climate variables. We tested for the presence of inter‐ and intraguild hotspots of endemism. Many individual plant and land snail taxa were tightly linked with geology, corresponding to small distributions. Conversely, most vertebrate taxa were not constrained to specific geological substrates and occurred over larger areas. However, across all three guilds climate was the strongest predictor of regional endemism, particularly for plants wherein discrete hotspots of endemism were buffered from extreme summer temperatures. Land snail and vertebrate endemism peaked in areas with highest precipitation in the driest times of the year. Hotspots of endemism within each guild poorly predicted endemism in other guilds. We found an overarching signal that climatic gradients play a dominant role in the persistence of endemic taxa in an arid‐zone mountain range system. An association with higher rainfall and cooler temperatures indicates that continuing trends toward hotter and drier climates may lead to range contractions in this, and potentially other, arid‐zone mountain biotas. Contrasting patterns of endemism across guilds highlight the need to couple comprehensive regional planning for the protection of climate refugia, with targeted management of more localized and habitat specialist taxa.     

BIOFRAG – a new database for analyzing BIOdiversity responses to forest FRAGmentation

Habitat fragmentation studies have produced complex results that are challenging to synthesize. Inconsistencies among studies may result from variation in the choice of landscape metrics and response variables, which is often compounded by a lack of key statistical or methodological information. Collating primary datasets on biodiversity responses to fragmentation in a consistent and flexible database permits simple data retrieval for subsequent analyses. We present a relational database that links such field data to taxonomic nomenclature, spatial and temporal plot attributes, and environmental characteristics. Field assessments include measurements of the response(s) (e.g., presence, abundance, ground cover) of one or more species linked to plots in fragments within a partially forested landscape. The database currently holds 9830 unique species recorded in plots of 58 unique landscapes in six of eight realms: mammals 315, birds 1286, herptiles 460, insects 4521, spiders 204, other arthropods 85, gastropods 70, annelids 8, platyhelminthes 4, Onychophora 2, vascular plants 2112, nonvascular plants and lichens 320, and fungi 449. Three landscapes were sampled as long-term time series (>10 years). Seven hundred and eleven species are found in two or more landscapes. Consolidating the substantial amount of primary data available on biodiversity responses to fragmentation in the context of land-use change and natural disturbances is an essential part of understanding the effects of increasing anthropogenic pressures on land. The consistent format of this database facilitates testing of generalizations concerning biologic responses to fragmentation across diverse systems and taxa. It also allows the re-examination of existing datasets with alternative landscape metrics and robust statistical methods, for example, helping to address pseudo-replication problems. The database can thus help researchers in producing broad syntheses of the effects of land use. The database is dynamic and inclusive, and contributions from individual and large-scale data-collection efforts are welcome.     

Woody plants and the prediction of climate-change impacts on bird diversity

Current methods of assessing climate-induced shifts of species distributions rarely account for species interactions and usually ignore potential differences in response times of interacting taxa to climate change. Here, we used species-richness data from 1005 breeding bird and 1417 woody plant species in Kenya and employed model-averaged coefficients from regression models and median climatic forecasts assembled across 15 climate-change scenarios to predict bird species richness under climate change. Forecasts assuming an instantaneous response of woody plants and birds to climate change suggested increases in future bird species richness across most of Kenya whereas forecasts assuming strongly lagged woody plant responses to climate change indicated a reversed trend, i.e. reduced bird species richness. Uncertainties in predictions of future bird species richness were geographically structured, mainly owing to uncertainties in projected precipitation changes. We conclude that assessments of future species responses to climate change are very sensitive to current uncertainties in regional climate-change projections, and to the inclusion or not of time-lagged interacting taxa. We expect even stronger effects for more specialized plant–animal associations. Given the slow response time of woody plant distributions to climate change, current estimates of future biodiversity of many animal taxa may be both biased and too optimistic.     

Does organic farming benefit farmland birds in winter?

The generally higher biodiversity on organic farms may be influenced by management features such as no synthetic pesticide and fertilizer inputs and/or by differences in uncropped habitat at the site and landscape scale. We analysed bird and habitat data collected on 48 paired organic and conventional farms over two winters to determine the extent to which broad-scale habitat differences between systems could explain overall differences in farmland bird abundance. Density was significantly higher on organic farms for six out of 16 species, and none on conventional. Total abundance of all species combined was higher on organic farms in both years. Analyses using an information-theoretic approach suggested that both habitat extent and farm type were important predictors only for starling and greenfinch. Organic farming as currently practised may not provide significant benefits to those bird species that are limited by winter food resources, in particular, several declining granivores.     

The value of woody hedgerows for moth diversity on organic and conventional farms

Habitat destruction and degradation are important drivers of biodiversity loss within agro-ecosystems. However, little is known about the effect of farming practices and the value of woody hedgerows on Lepidoptera in North America. The purpose of this work was to study moth diversity in woody hedgerows and croplands of organic and conventional farms. In addition, the influence of vegetation composition and abiotic variables on species richness, abundance, and composition was examined. Moths were sampled with light traps during six weeks in the summer of 2001. Vegetation data and abiotic variables were obtained for all sites. In total, 26,020 individuals from 12 families and 408 species were captured. Most species were uncommon. Only 35 species included >100 individuals while for 71% of species <10 individuals were found. The Noctuidae represented 221 species and 85% of all individuals captured. Woody hedgerows harbored more species and in greater number than croplands. There was no significant difference in moth diversity between organic and conventional farms, except that the Notodontidae were significantly more species rich in organic than in conventional sites. Results show that species richness, abundance, and composition were greatly influenced by habitat types (hedgerow versus crop field) and abiotic variables (minimum temperature which was correlated to moon illumination, rainfall, and cloud cover). Moth species composition was significantly correlated to vegetation composition. This study broadens our understanding of the factors driving moth diversity and expands our knowledge of their geographic range. The maintenance of noncrop habitats such as woody hedgerows within agro-ecosystems seems paramount to preserving the biodiversity and abundance of many organisms, including moths.     

No escape: The influence of substrate sodium on plant growth and tissue sodium responses

1. As an essential micronutrient for many organisms, sodium plays an important role in ecological and evolutionary dynamics. Although plants mediate trophic fluxes of sodium, from substrates to higher trophic levels, relatively little comparative research has been published about plant growth and sodium accumulation in response to variation in substrate sodium. Accordingly, we carried out a systematic review of plants'' responses to variation in substrate sodium concentrations.
2. We compared biomass and tissue‐sodium accumulation among 107 cultivars or populations (67 species in 20 plant families), broadly expanding beyond the agricultural and model taxa for which several generalizations previously had been made. We hypothesized a priori response models for each population''s growth and sodium accumulation as a function of increasing substrate NaCl and used Bayesian Information Criterion to choose the best model. Additionally, using a phylogenetic signal analysis, we tested for phylogenetic patterning of responses across taxa.
3. The influence of substrate sodium on growth differed across taxa, with most populations experiencing detrimental effects at high concentrations. Irrespective of growth responses, tissue sodium concentrations for most taxa increased as sodium concentration in the substrate increased. We found no strong associations between the type of growth response and the type of sodium accumulation response across taxa. Although experiments often fail to test plants across a sufficiently broad range of substrate salinities, non‐crop species tended toward higher sodium tolerance than domesticated species. Moreover, some phylogenetic conservatism was apparent, in that evolutionary history helped predict the distribution of total‐plant growth responses across the phylogeny, but not sodium accumulation responses.
4. Our study reveals that saltier plants in saltier soils proves to be a broadly general pattern for sodium across plant taxa. Regardless of growth responses, sodium accumulation mostly followed an increasing trend as substrate sodium levels increased.