Multiple Euryhaline Lineages of Centrohelids (Haptista: Centroplasthelida) in Inland Saline Waters Revealed with Metabarcoding

The diversity of centrohelids in inland saline waters was studied with metabarcoding for the first time. The fragment of V6–V7 regions of 18S rDNA was sequenced with newly designed primers. Obtained OTUs were identified with molecular phylogenetic analysis and comparison of the signatures in 39es9 hairpin of V7. The obtained data included some OTUs, which could be attributed to four described species, but the majority belonged to previously established or novel environmental clades. Along with some presumably marine/brackish clades and freshwater/low salinity (0–2 ppt) clades, seven presumable species demonstrating broad (from 1–2 up to 78 ppt) salinity tolerance were detected. A number of OTUs belonged to Raphidocystis contractilis, which is known from three independent findings in brackish habitats only. Thus, it was assumed that this species is stenohaline and specifically adapted to salinity 5–15 ppt. The high level of salinity tolerance was suggested for centrohelids before based on morphology, which was used to justify their cosmopolitan distribution. Later these views were criticized based on environmental sequencing, but the results of the current survey indicate, that at least some species are present at salinities from almost freshwater (1–2 ppt) to twice oceanic (78 ppt) and are presumably capable of overcoming oceanic salinity barriers for their distribution.     

The effects of salinity and temperature on the development and survival of fish parasites

In brackish water the variety of marine and freshwater parasite species is considerably reduced. The distribution in brackish water of most marine endoparasites is restricted by the salinity tolerance of their hosts, most of the parasite species are more tolerant than their hosts. The influence of salinity and temperature on nine species has been examined; first stage larvae of Contracaecum aduncum develop in 0-32‰ salinity; Cryptocotyle lingua proved to be infective at salinities down to 4‰. The greatest resistance was found in Anisakis larvae from herring Clupea harengus , which survived for more than half a year. Parasites in the fish intestines appear to be unaffected by changing water salinities, as the osmolarity in the intestines stays nearly constant. Marine ectoparasites ( Acanthochondria depressa, Lepeophtheirus pectoralis ) survive about three times longer than freshwater species ( Piscicola geometra, Argulus foliaceus ) when salinity is 16‰. High temperature increases the effects of adverse salinities on parasites. There is evidence that none of these ecto-parasitic species can develop within the range of 7-20‰ salinity.     

Testing for beneficial reversal of dominance during salinity shifts in the invasive copepod Eurytemora affinis,and implications for the maintenance of genetic variation

Maintenance of genetic variation at loci under selection has profound implications for adaptation under environmental change. In temporally and spatially varying habitats, non‐neutral polymorphism could be maintained by heterozygote advantage across environments (marginal overdominance), which could be greatly increased by beneficial reversal of dominance across conditions. We tested for reversal of dominance and marginal overdominance in salinity tolerance in the saltwater‐to‐freshwater invading copepod Eurytemora affinis. We compared survival of F1 offspring generated by crossing saline and freshwater inbred lines (between‐salinity F1 crosses) relative to within‐salinity F1 crosses, across three salinities. We found evidence for both beneficial reversal of dominance and marginal overdominance in salinity tolerance. In support of reversal of dominance, survival of between‐salinity F1 crosses was not different from that of freshwater F1 crosses under freshwater conditions and saltwater F1 crosses under saltwater conditions. In support of marginal overdominance, between‐salinity F1 crosses exhibited significantly higher survival across salinities relative to both freshwater and saltwater F1 crosses. Our study provides a rare empirical example of complete beneficial reversal of dominance associated with environmental change. This mechanism might be crucial for maintaining genetic variation in salinity tolerance in E. affinis populations, allowing rapid adaptation to salinity changes during habitat invasions.     

Tolerance to salinity and thermal stress by larvae and adults of the serpulid annelid Ficopomatus enigmaticus

The serpulid annelid Ficopomatus enigmaticus is a widely distributed invader of shallow‐water, brackish habitats in subtropical and temperate regions, where it has numerous damaging ecological and economic effects. Its distributional pattern suggests that temperature and salinity play important roles in limiting its distribution, but because other factors often covary with these, drawing strong conclusions from these patterns is difficult. In an effort to more clearly identify the effects of these factors, we examined tolerance to acute thermal (16–28°C) and salinity (0–35 psu) stress by larvae (5‐day exposure, unfed) and adults (14‐day exposure, unfed) of F. enigmaticus in the laboratory experiments. Larvae showed higher mortality at the highest temperature tested 28°C; adult survival was unaffected by temperature. Neither larvae nor adults survived exposure to pure freshwater (0 psu), but survived well at salinities ranging 3.5–35 psu. In addition, high salinity did not slow tube growth in adults. These results suggest that salinity stress, in particular, does not directly limit the distribution of F. enigmaticus to low‐salinity habitats. Experimental work on the distribution of F. enigmaticus is uncommon in the literature, but is likely needed to identify the abiotic or biotic factors that limit the distribution of this frequently invasive species.     

Influence of density and salinity on larval development of salt‐adapted and salt‐naïve frog populations

Environmental change and habitat fragmentation will affect population densities for many species. For those species that have locally adapted to persist in changed or stressful habitats, it is uncertain how density dependence will affect adaptive responses. Anurans (frogs and toads) are typically freshwater organisms, but some coastal populations of green treefrogs (Hyla cinerea) have adapted to brackish, coastal wetlands. Tadpoles from coastal populations metamorphose sooner and demonstrate faster growth rates than inland populations when reared solitarily. Although saltwater exposure has adaptively reduced the duration of the larval period for coastal populations, increases in densities during larval development typically increase time to metamorphosis and reduce rates of growth and survival. We test how combined stressors of density and salinity affect larval development between salt‐adapted (“coastal”) and nonsalt‐adapted (“inland”) populations by measuring various developmental and metamorphic phenotypes. We found that increased tadpole density strongly affected coastal and inland tadpole populations similarly. In high‐density treatments, both coastal and inland populations had reduced growth rates, greater exponential decay of growth, a smaller size at metamorphosis, took longer to reach metamorphosis, and had lower survivorship at metamorphosis. Salinity only exaggerated the effects of density on the time to reach metamorphosis and exponential decay of growth. Location of origin affected length at metamorphosis, with coastal tadpoles metamorphosing slightly longer than inland tadpoles across densities and salinities. These findings confirm that density has a strong and central influence on larval development even across divergent populations and habitat types and may mitigate the expression (and therefore detection) of locally adapted phenotypes.     

The phytosociology of British sea-cliff vegetation with special reference to the ecophysiology of some maritime cliff plants

The major factors thought to control the distribution of associations on the sea cliffs in Britain are discussed in relation to the zonation of cliff vegetation, and some experiments to investigate the effects of these factors are described.The seeds of the maritime cliff species are able to germinate in higher salinities than those of closely related inland species.Relative growth rates of maritime cliff species indicate stimulation at low salinities over non-saline conditions, and less reduction in growth at higher salinities than those of closely related inland species.Increasing salinity reduces both photosynthesis and dark respiration in Lavatera arborea. Mesophyll and stomatal resistances are increased while transpiration is reduced.The distribution of ions within Lavatera arborea grown at different salinities indicates differential accumulation between young leaves, old leaves and roots.Nomenclature follows Clapham, Tutin & Warburg (1962).     

Broad Salinity Tolerance as a Refuge from Predation in the Harmful Raphidophyte Alga Heterosigma akashiwo (Raphidophyceae)

The ability of harmful algal species to form dense, nearly monospecific blooms remains an ecological and evolutionary puzzle. We hypothesized that predation interacts with estuarine salinity gradients to promote blooms of Heterosigma akashiwo (Y. Hada) Y. Hada ex Y. Hara et M. Chihara, a cosmopolitan toxic raphidophyte. Specifically, H. akashiwo's broad salinity tolerance appears to provide a refuge from predation that enhances the net growth of H. akashiwo populations through several mechanisms. (1) Contrasting salinity tolerance of predators and prey. Estuarine H. akashiwo isolates from the west coast of North America grew rapidly at salinities as low as six, and distributed throughout experimental salinity gradients to salinities as low as three. In contrast, survival of most protistan predator species was restricted to salinities >15. (2) H. akashiwo physiological and behavioral plasticity. Acclimation to low salinity enhanced H. akashiwo's ability to accumulate and grow in low salinity waters. In addition, the presence of a ciliate predator altered H. akashiwo swimming behavior, promoting accumulation in low‐salinity surface layers inhospitable to the ciliate. (3) Negative effects of low salinity on predation processes. Ciliate predation rates decreased sharply at salinities <25 and, for one species, H. akashiwo toxicity increased at low salinities. Taken together, these behaviors and responses imply that blooms can readily initiate in low salinity waters where H. akashiwo would experience decreased predation pressure while maintaining near‐maximal growth rates. The salinity structure of a typical estuary would provide this HAB species a unique refuge from predation. Broad salinity tolerance in raphidophytes may have evolved in part as a response to selective pressures associated with predation.     

Origins and functional diversification of salinity‐responsive Na+, K+ ATPase α1 paralogs in salmonids

The Salmoniform whole‐genome duplication is hypothesized to have facilitated the evolution of anadromy, but little is known about the contribution of paralogs from this event to the physiological performance traits required for anadromy, such as salinity tolerance. Here, we determined when two candidate, salinity‐responsive paralogs of the Na⁺, K⁺ ATPase α subunit (α1a and α1b) evolved and studied their evolutionary trajectories and tissue‐specific expression patterns. We found that these paralogs arose during a small‐scale duplication event prior to the Salmoniform, but after the teleost, whole‐genome duplication. The ‘freshwater paralog’ (α1a) is primarily expressed in the gills of Salmoniformes and an unduplicated freshwater sister species (Esox lucius) and experienced positive selection in the freshwater ancestor of Salmoniformes and Esociformes. Contrary to our predictions, the ‘saltwater paralog’ (α1b), which is more widely expressed than α1a, did not experience positive selection during the evolution of anadromy in the Coregoninae and Salmonine. To determine whether parallel mutations in Na⁺, K⁺ ATPase α1 may contribute to salinity tolerance in other fishes, we studied independently evolved salinity‐responsive Na⁺, K⁺ ATPase α1 paralogs in Anabas testudineus and Oreochromis mossambicus. We found that a quarter of the mutations occurring between salmonid α1a and α1b in functionally important sites also evolved in parallel in at least one of these species. Together, these data argue that paralogs contributing to salinity tolerance evolved prior to the Salmoniform whole‐genome duplication and that strong selection and/or functional constraints have led to parallel evolution in salinity‐responsive Na⁺, K⁺ ATPase α1 paralogs in fishes.     

Ecological correlates of the distribution limits of two poeciliid species along a salinity gradient

Identifying the environmental factors responsible for the formation of a species' distribution limit is challenging because organisms interact in complex ways with their environments. However, the use of statistical niche models in combination with the analysis of phenotypic variation along environmental gradients can help to reduce such complexity and identify a subset of candidate factors. In the present study, we used such approaches to describe and identify factors responsible for the parapatric distribution of two closely‐related livebearer fish species along a salinity gradient in the lowlands of Trinidad, West Indies. The downstream distribution limits of Poecilia reticulata were strongly correlated with the brackish–freshwater interface. We did not observe significant phenotypic variation in life‐history traits for this species when comparing marginal with more central populations, suggesting that abrupt changes in conditions at the brackish–freshwater interface limit its distribution. By contrast, Poecilia picta was present across a wide range of salinities, although it gradually disappeared from upstream freshwater localities. In addition, P. picta populations exhibited an increase in offspring size in localities where they coexist with P. reticulata, suggesting a role for interspecific competition. The parapatric distribution of these two species, suggests that P. reticulata distributions are limited by an abiotic factor (salinity), whereas P. picta is limited by a biotic factor (interspecific competition). Similar parapatric patterns have been previously described in other systems, suggesting they might be a common pattern in nature. © 2013 The Linnean Society of London     

Comparative survival and growth of embryos,larvae, and juveniles of pejerrey Odontesthes bonariensis and O. hatcheri* at different salinities

The hatching of fertilized eggs and the survival and growth of larvae and juveniles of the inland‐water atherinids Odontesthes bonariensis (Valenciennes 1835) and O. hatcheri (Eigenmann 1909) were examined at salinities of 0, 5, 10, 20, and 30 ppt. In addition, a limited study compared the salinity responses of O. bonariensis eggs and larvae from different origins. Overall, embryos, larvae, and juveniles of both species were euryhaline, although best survival and growth rates were obtained at the intermediate salinities. Survival of O. bonariensis at 0 ppt varied from very good to very poor. Comparison of the salinity responses of eggs and larvae of O. bonariensis from the current Japanese strain with newly introduced strains from three locations in Argentina did not reveal a clearly superior strain for freshwater culture. In general, O. hatcheri showed higher survival and growth rates and better adaptability to fresh water compared with O. bonariensis. Although both species are commonly regarded as freshwater species, the results of this study emphasize the importance of millimolar quantities of salts in the rearing water for improved survival and growth.     

Evolution of salinity tolerance in the diving beetle tribe Hygrotini (Coleoptera,Dytiscidae)

Some species of the diving beetle tribe Hygrotini (subfamily Hydroporinae) are among the few insects able to tolerate saline concentrations more than twice that of seawater. However, the phylogenetic relationships of the species of Hygrotini, and the origin and evolution of tolerance to salinity in this lineage, are unknown. In this work, we aim to reconstruct how many times salinity tolerance did evolve in Hygrotini, whether this evolution was gradual or if tolerance to hypersalinity could evolve directly from strictly freshwater (FW) species, and to estimate the probabilities of transition between habitats. We build a phylogeny with ca. 45% of the 137 species of Hygrotini, including all major lineages and almost all of the known halophile or tolerant species. We used sequence data of four mitochondrial (COI‐5′, COI‐3′, 16S + tRNA and NADH1) and three nuclear (28S, 18S and H3) gene fragments, plus ecological data to reconstruct the history of the salinity tolerance using Bayesian inference. Our results demonstrate multiple origins of the tolerance to salinity, although most saline and hypersaline species were concentrated in two lineages. The evolution of salinity was gradual, with no direct transitions from FW to hypersaline habitats, but with some reversals from tolerant to FW species. The oldest transition to saline tolerance, at the base of the clade with the highest number of saline species, was dated in the late Eocene‐early Oligocene, a period with decreasing temperature and precipitation. This temporal coincidence suggests a link between increased aridity and the development of tolerance to saline waters, in agreement with recent research in other groups of aquatic Coleoptera.     

Ecophysiological adaptability of tropical water organisms to salinity changes]

Physiological response of tropical organisms to salinity changes was studied for some marine, estuarine and freshwater fishes (Astyanax bimaculatus, Petenia karussii, Cyprinodon dearborni, and Oreochromis mossambicus), marine and freshwater crustaceans (Penaeus brasiliensis, Penaeus schmitti and Macrobrachium carcinus), and marine bivalves (Perna perna, Crassostrea rhizophorae, and Arca zebra) collected from Northeast Venezuela. They were acclimated for four weeks at various salinities, and (1) placed at high salinities to determine mean lethal salinity, (2) tested by increasing salinity 5@1000 per day to define upper lethal salinity tolerance limit, or (3) observed in a saline gradient tank to determine salinity preference. Acclimation level was the most significant factor. This phenomenon is important for tropical aquatic organisms in shallow waters, where they can adapt to high salinity during the dry season and cannot lose their acclimation level at low salinity during abrupt rain. For saline adaptation of tropical organisms, this behavior will contribute to their proliferation and distribution in fluctuating salinity environments.     

Influence of Salinity Concentration on Aquatic Insect Community Structure: A Mesocosm Experiment in the Dead Sea Basin Region

Salinity varies considerably among temporary pools in the Dead Sea Basin, Israel. We experimentally assessed the effects of four salinity levels (0, 10, 20 and 30 g NaCl per liter) on the aquatic insect community in this basin in an artificial pool experiment. Each salinity level was randomly assigned to six pools (total=24 pools). Salinity did not affect total insect abundance but strongly affected abundance and distributions of different species, and consequently, community structure. Of 13 taxa colonizing the pools, 12 were Diptera including 10 mosquito species. Five taxa were sufficiently common to assess abundance in relation to salinity. Polypedilum nubiferum Skuse (Diptera: Chironomidae) was largely salinity intolerant being abundant only in the freshwater. Ephydra flavipes Macquart (Diptera: Ephydridae) was most abundant at the highest salinity level and was rare in freshwater. Ochlerotatus caspius Pallas (Diptera: Culicidae) abundance tended to be highest at 10 g/l and lowest at 30 g/l although the differences were not statistically significant. Anopheles multicolor Cambouliu (Diptera: Culicidae) was relatively euryhaline although numbers dropped significantly at the highest salinity. Cleon dipterum Linnaeus (Baetidae: Ephemeroptera) was also euryhaline and showed no significant differences in abundance across salinities. For the mosquito species, we also estimated survival to pupation. Survival to pupation was significantly lower for O. caspius in freshwater, but was not statistically significantly different across salinities for A.␣multicolor. Species diversity was highest at the two lowest salinities tested and then dropped with increasing salinity. Evenness was not significantly different across salinities. Community similarity generally decreased with increasing salinity differences though dissimilarity was greatest when comparing freshwater to other salinities. Thus, regional diversity is likely increased when there is a range of salinities among pools.     

Effects of salinity on the growth and morphology of the invasive,euryhaline hydroid Cordylophora (Phylum Cnidaria,Class Hydrozoa)

The invasive, euryhaline hydroid Cordylophora sp. is a colonial cnidarian present in both freshwater and brackish water habitats. Individuals contend with osmotic stress at the tissue and cellular level. It has been suggested that this hydroid's ability to expand its range of distribution by invading new habitats is due in large part to an ability to acclimate to new salinities. The purpose of this study was to assess colony growth and morphological changes at various salinities in freshwater and brackish genotypes of Cordylophora sp. Single genotypes from a known freshwater clade (0.5 psu; Des Plaines River) and a known brackish clade (16 psu; Napa River) were cultured and gradually transitioned to 12 different salinities ranging 0.5–22 psu, and we characterized the growth rates and hydranth morphological features at each salinity. Colony growth was optimal at 0.5 psu for the freshwater genotype and 10 psu for the brackish genotype. Changes in hydranth morphology in the freshwater genotype were primarily observed at higher salinities, while morphological changes in the brackish genotype primarily occurred at lower salinities. Our results for the brackish genotype generally concur with previous work, but this study is the first to document the response of a freshwater genotype of Cordylophora sp. to various salinities. Differences in growth between these two genotypes strongly support the previously proposed existence of multiple cryptic species. Furthermore, because this hydroid is quite prevalent in freshwater and brackish systems as a fouling organism, understanding the effects of various salinities on the successful establishment of Cordylophora sp. is an important contribution to the understanding of the ecophysiology and management of this invasive hydroid.     

Reduced salinities compromise the thermal tolerance of hypersaline specialist diving beetles

Inland saline waters are globally threatened habitats that harbour unique assemblages of specialist invertebrates. In many Mediterranean regions, irrigation associated with intensive agriculture is lowering the salinity of these habitats, resulting in the loss of their specialist biota, although the mechanisms by which reductions in salinity lead to species loss are poorly understood. In the present study, the effects of reduced salinity on the temperature tolerance and thermal acclimatory abilities of two related species of hypersaline water beetles, Nebrioporus baeticus (Schaum) and Nebrioporus ceresyi (Aubé), are explored. Both upper (UTL) and lower thermal limits (LTL) are assessed, and both salinity and temperature are found to influence the thermal biology of Nebrioporus. Mean UTLs are greater in individuals of both species acclimated at high salinities, with salinity appearing to be more important than acclimation temperature in determining UTL. In both taxa, the lowest mean LTLs are recorded in individuals acclimated at the highest salinities and lowest temperatures; temperature‐dependent acclimation is only reported after exposure to relatively high salinities. The data show that salinity influences the thermal tolerance and acclimatory ability of these hypersaline beetles, and that lowered salinity compromises the ability of adult Nebrioporus to cope with both heat and cold. Such an effect may partly explain why specialist species are lost from hypersaline habitats subject to salinity reductions, and suggests that ongoing reduction in salinity may compromise the ability of such specialist taxa to cope with rapid climate change.     

Evolution of physiological tolerance and performance during freshwater invasions

Invasive species that penetrate habitat boundaries are likelyto experience strong selection and rapid evolution. This studydocuments evolutionary shifts in tolerance and performance followingthe invasion of fresh water by the predominantly estuarine andsalt marsh copepod Eurytemora affinis. Common-garden experimentswere performed on freshwater-invading (Lake Michigan) and ancestralsaline (St. Lawrence marsh) populations to measure shifts inadult survival (at 0, 5, and 25 PSU), and survival during developmentand development time (both using full-sib clutches split across0, 5, 15, and 25 PSU). Results showed clear evidence of heritableshifts in tolerance and performance associated with freshwaterinvasions. The freshwater population exhibited a gain in low-salinitytolerance and a reduction in high-salinity tolerance relativeto the saline population, suggesting tradeoffs. These tradeoffswere supported by negative genetic correlations between survivalat fresh (0 PSU) versus higher salinities. Mortality in responseto salinity occurred primarily before metamorphosis, suggestingthat selection in response to salinity had acted primarily onthe early life-history stages. The freshwater population exhibitedcurious patterns of life-history evolution across salinities,relative to the saline population, of retarded development tometamorphosis but accelerated development from metamorphosisto adulthood. This pattern might reflect tradeoffs between developmentrate and survival in fresh water at the early life-history stages,but some other selective force acting on later life-historystages. Significant effects of clutch (genotype) and clutch-by-salinityinteraction (G x E) on survival and development time in bothpopulations indicated ample genetic variation as substrate fornatural selection. Variation for high-salinity tolerance waspresent in the freshwater population despite negative geneticcorrelations between high- and low-salinity tolerance. Resultsimplicate the importance of natural selection and document theevolution of reaction norms during freshwater invasions.     

Characterization of halotolerant Bicosoecida and Placididea (Stramenopila) that are distinct from marine forms,and the phylogenetic pattern of salinity preference in heterotrophic stramenopiles

Recent culture‐based studies demonstrate the distinctiveness of the microbial eukaryote biota of very hypersaline environments. In contrast, microscopy‐based faunistic studies suggest that the biota of habitats of more moderate hypersalinity (60–150‰) overlaps substantially with that of marine environments, but this has barely been studied with modern techniques. To investigate the diversity and salinity tolerance range of these organisms, eight cultures of heterotrophic stramenopiles were established from (or from nearby) moderately hypersaline locations. These isolates represent five independent groups; Groups A, B and C are bicosoecids; Groups D and E belong to Placididea. One isolate (Group A) is a strain of the widespread marine species Cafeteria roenbergensis, and cannot grow above 100‰ salinity. The other isolates – Groups B–E – can all grow at 150–175‰ salinities and are probably moderate halophiles. Groups B–E all represent previously unsequenced species or even genera, although Group B is the sister group of the borderline extreme halophile Halocafeteria. The high level of novelty en countered suggests that moderately hypersaline environments may harbour a heterotrophic stramenopile biota distinct from that of marine environments. Interestingly, our new isolates are all most closely related to marine or halophilic forms, and our phylogenies show large clades defined by saline/non‐saline habitats within bicosoecids, placidomonads and related lineages. In particular, most freshwater/soil bicosoecids form one well‐supported clade. The sole major exception is Bicosoeca, which is intermixed with marine environmental sequences originally referred to as ‘MAST‐13’, which are from brackish water, not typical seawater. It seems that the freshwater/marine barrier has been crossed very few times in the evolutionary history of these heterotrophic stramenopile flagellates.     

Salinity tolerance of diapausing eggs of freshwater zooplankton

1. Many freshwater zooplankton produce diapausing eggs capable of withstanding periods of adverse environmental conditions, such as anoxia, drought and extreme temperature. These eggs may also allow oligostenohaline species to survive increased salinity during periods of tidal flux or evaporation, and here we test the ability of diapause eggs to withstand such conditions. 2. Salinity tolerance may also enable organisms to invade new environments. The increased rate of introduction of non‐indigenous species to the Laurentian Great Lakes since 1989, when ballast water exchange regulations (to replace fresh/brackish water at sea with full seawater) were first implemented for transoceanic vessels, has stimulated studies that explore mechanisms of introduction, other than of active animals, in ballast water. One hypothesis proposes that freshwater organisms transported in ballast tanks as diapausing eggs may be partially responsible for the increased rate of species introduction, as these eggs may tolerate a wide array of adverse environmental conditions, including exposure to saline water. 3. We collected ballast sediments from transoceanic vessels entering the Great Lakes, isolated diapausing eggs of three species (Bosmina liederi, Daphnia longiremis and Brachionus calyciflorus), and measured the effect of salinity on hatching rate. In general, exposure to salinity significantly reduced the hatching rate of diapausing eggs. However, as non‐indigenous species can establish from a small founding population, it is unclear whether salinity exposure will be effective as a management tool.