Two modified Breder traps for quantitative studies of tropical amphidromous migration

Amphidromy is a life cycle of fauna found throughout the tropics and subtropics, including representatives from three phyla. Amphidromy involves a life cycle of oceanic larval development, with postlarval migration into streams where growth and reproduction take place. With increasing industrial and urban development in tropical regions, demands for freshwater are continuously growing, resulting in the construction of dams and diversions that break the freshwater continuum necessary for the amphidromous life cycle; thus, jeopardizing many populations of native and endemic tropical stream fauna. Because of this, more quantitative studies are needed to better understand this unique life cycle, which heretofore, has been relatively little studied compared to tropical terrestrial communities. As such, new and quantitative methods are needed to study the immature stages (e.g., postlarvae) associated with amphidromy. In this paper, we introduce two modifications of the original Breder trap that have been designed specifically for standardized, quantitative monitoring of amphidromous postlarval migration. In addition, the two modified traps can be used in a variety of stream settings from natural bed substrates to modified channels with little heterogeneity. The first modification is made of acrylic, with a flat bottom useful in channelized streams or streams with relatively flat, unnatural benthic substrate (e.g., concrete). The second modification is an affordable trap made from polyvinylchloride (PVC) compression couplings, to be used in streams with natural benthic substrates, particularly those with large, embedded and immovable boulders and bedrock outcrops. Both traps were designed for continuous water flow through the traps, providing the necessary rheotactic cue for migrating amphidromous postlarvae, and ameliorating deficiencies of earlier traps not intended for tropical amphidromous fauna. We also make recommendations for standardized use of these traps to facilitate data comparisons among studies.     

Two modified Breder traps for quantitative studies of tropical amphidromous migration

Amphidromy is a life cycle of fauna found throughout the tropics and subtropics, including representatives from three phyla. Amphidromy involves a life cycle of oceanic larval development, with postlarval migration into streams where growth and reproduction take place. With increasing industrial and urban development in tropical regions, demands for freshwater are continuously growing, resulting in the construction of dams and diversions that break the freshwater continuum necessary for the amphidromous life cycle; thus, jeopardizing many populations of native and endemic tropical stream fauna. Because of this, more quantitative studies are needed to better understand this unique life cycle, which heretofore, has been relatively little studied compared to tropical terrestrial communities. As such, new and quantitative methods are needed to study the immature stages (e.g., postlarvae) associated with amphidromy. In this paper, we introduce two modifications of the original Breder trap that have been designed specifically for standardized, quantitative monitoring of amphidromous postlarval migration. In addition, the two modified traps can be used in a variety of stream settings from natural bed substrates to modified channels with little heterogeneity. The first modification is made of acrylic, with a flat bottom useful in channelized streams or streams with relatively flat, unnatural benthic substrate (e.g., concrete). The second modification is an affordable trap made from polyvinylchloride (PVC) compression couplings, to be used in streams with natural benthic substrates, particularly those with large, embedded and immovable boulders and bedrock outcrops. Both traps were designed for continuous water flow through the traps, providing the necessary rheotactic cue for migrating amphidromous postlarvae, and ameliorating deficiencies of earlier traps not intended for tropical amphidromous fauna. We also make recommendations for standardized use of these traps to facilitate data comparisons among studies.     

Migratory drift of larval freshwater shrimps in two tropical streams, Puerto Rico

1. Migratory shrimps are often major biotic components of tropical stream communities, yet spatial and temporal patterns of their migration have yet to be described. This information is of increasing importance given the continued fragmentation of tropical streams by damming and water abstraction/diversion, which can disrupt migratory life cycles. 2. Larval amphidromous shrimps are released by adult females in freshwater streams. They then drift passively to an estuarine habitat where they metamorphose before migrating back upstream. Drift of larval shrimps was sampled over two to five 24-h periods at each of three sites along two rivers that drain the Luquillo Experimental Forest in Puerto Rico: the Espíritu Santo (10, 135 and 335 m a.s.l.) and the Mameyes (10, 90 and 290 m a.s.l.). A total of seventeen diel samplings were conducted. 3. Shrimp drift increased in the downstream direction in both catchments, and had a significant positive exponential relationship with length of stream channel above each site. There was no significant difference between catchments with respect to mean daily drift rate per km of stream channel. Maximum observed larval shrimp density was 69 102 larvae 100 m^–3 (1.7 g dry mass 100 m^–3), which is high relative to published invertebrate drift studies. 4. The pattern of shrimp drift agreed with the ’risk of predation hypothesis‘. In stream reaches with predatory fish, drift of larval shrimps occurred at night and was slight during the day. A nocturnal peak in drift occurred between 19.00 and 22.00 h. At a high-altitude site, where predatory fish were absent, no diel pattern was discernible. 5. The present study provides information on the timing of migratory drift of larval shrimps, which can minimize the adverse effects of water abstraction from streams draining the Luquillo Experimental Forest. Elimination of water withdrawal during peak larval drift after dark will significantly reduce shrimp mortality.     

Experimental evaluation of a minnow trap for small lotic fish

A minnow trap that operates in various flow regimes in streams and allows sampling of small fish from stream bed microhabitats was developed. In laboratory and field tests, the most efficient trap design for capturing and retaining various species of fish had one funnel oriented downstream, a plexiglass body, and commercial trout food as bait. These lightweight traps can be set in a wide range of current velocities and depths, and can be useful in investigations that examine the microhabitat use, diel activity patterns or population densities of small lotic fish. Guidelines for the trap's use and for quick verification of capture success in new situations are suggested.     

Amphidromy links a newly documented fish community of continental Australian streams, to oceanic islands of the west Pacific

Background

Indo-Pacific high island streams experience extreme hydrological variation, and are characterised by freshwater fish species with an amphidromous life history. Amphidromy is a likely adaptation for colonisation of island streams following stochastic events that lead to local extirpation. In the Wet Tropics of north-eastern Australia, steep coastal mountain streams share similar physical characteristics to island systems. These streams are poorly surveyed, but may provide suitable habitat for amphidromous species. However, due to their ephemeral nature, common non-diadromous freshwater species of continental Australia are unlikely to persist. Consequently, we hypothesise that coastal Wet Tropics streams are faunally more similar, to distant Pacific island communities, than to nearby faunas of large continental rivers.

Methods/Principal Findings

Surveys of coastal Wet Tropics streams recorded 26 species, 10 of which are first records for Australia, with three species undescribed. This fish community is unique in an Australian context in that it contains mostly amphidromous species, including sicydiine gobies of the genera Sicyopterus, Sicyopus, Smilosicyopus and Stiphodon. Species presence/absence data of coastal Wet Tropics streams were compared to both Wet Tropics river networks and Pacific island faunas. ANOSIM indicated the fish fauna of north-eastern Australian coastal streams were more similar to distant Pacific islands (R = 0.76), than to nearby continental rivers (R = 0.98).

Main Conclusions/Significance

Coastal Wet Tropics streams are faunally more similar to distant Pacific islands (79% of species shared), than to nearby continental fauna due to two factors. First, coastal Wet Tropics streams lack many non-diadromous freshwater fish which are common in nearby large rivers. Second, many amphidromous species found in coastal Wet Tropics streams and Indo-Pacific islands remain absent from large rivers of the Wet Tropics. The evolutionary and conservation significance of this newly discovered Australian fauna requires clarification in the context of the wider amphidromous fish community of the Pacific.     

Similarities in Behavioral Ecology among Amphidromous and Catadromous Fishes on the Oceanic Islands of Hawai'i and Guam

About seven families of fishes occur routinely in fresh water on oceanic high islands of the tropical Pacific; others (sharks, jacks, bonefish, etc.) are occasional visitors. However, amphidromous fishes (freshwater adults, marine larvae) of the families Gobiidae and Eleotridae are predominant in island streams. Hawai'i, representing the northernmost extent of Polynesia, has five species of gobioid fishes whose adults are limited to fresh water, but Guam, in the Mariana Islands of the far Western Pacific, has more than four times that number. Hawaiian stream fishes are strikingly similar to their Guamanian relatives in their distribution, ecology, and behavior. At both localities, these fishes typically exhibit strong species specificity in the section of stream inhabited by adults, in the microhabitat selected, and in their food and feeding. Although incompletely understood, aspects of the life cycles of amphidromous island fishes (spawning, migrations into and from the sea, and others) are cued by seasonal and short-term changes in stream flow. In the Hawaiian Islands, water-use decisions based on the imperatives of allowing no net loss of habitat for aquatic animals and maintaining stream-ocean pathways for migrating animals have facilitated both management and conservation of diversity in island streams.     

Rapid acquisition of directional preferences by migratory juveniles of two amphidromous Hawaiian gobies, Awaous guamensis and Sicyopterus stimpsoni

The native freshwater fish fauna of the Hawaiian Islands is composed of four gobiid species and one eleotrid. All five species are amphidromous. They spawn in freshwater, go to sea as free embryos and, as juveniles (sometimes referred to as post-larvae), enter fresh water again. Juveniles of two of these species, Awaous guamensis and Sicyopterus stimpsoni, had been observed maintaining directed migratory activity in the absence of water flow during the early freshwater phase of their upstream migration. Our experiments demonstrate that these juveniles are strongly attracted to flowing water. But, when water flow stops the fish persist in migratory behaviour in the same compass direction even when the tank is rotated, controlling for local landmarks and chemical cues, or when the trap that was the original water source is moved. This indicates a rapid (10 min or less) learning of directional cues. Preliminary data indicates a similar ability in a third species, Lentipes concolor. Rapid acquisition of directional preferences may be critical during upstream migration, when these tiny fish traverse water falls and other barriers by climbing in intermittent water flows. We suggest that rapid directional learning may have evolved in gobies as part of their predator avoidance repertoire and been subsequently applied to migration in streams.     

Why be amphidromous: expatrial dispersal and the place of source and sink population dynamics?

Amphidromous fishes are found predominantly on the tropical and subtropical islands of the globe and there are few amphidromous species on continents. I suggest that this idiosyncratic distribution relates in part to problems in self-recruitment on islands that are often young or volcanic, and which may have streams with ephemeral flows across relatively short times scales. Amphidromy provides the ability to invade new habitats as these become available either on newly emergent (often volcanic) islands, or following perturbation after stream dewatering or the impacts of volcanism on older islands as a consequence of expatrial dispersal. Source/sink population dynamics may also be involved with islands ‘downstream’ in oceanic current systems behaving as sinks, with little or no self-recruitment. Streams in steep topography seem to be favoured by amphidromous species, perhaps because they provide more rapid transport to sea of the tiny, newly hatched larvae.     

Dispersal and upstream migration of an amphidromous neritid snail: implications for restoring migratory pathways in tropical streams

1. The amphidromous life cycle of several species of neritid snails, shrimp and gobies throughout the tropics includes juveniles that migrate from the ocean to breed in fresh water. In many Hawaiian streams, the decline of Neritina granosa, an endemic gastropod, has been associated with habitat degradation and water withdrawal, which are common factors affecting tropical rivers around the world. 2. We investigated the effects of water withdrawal and density on dispersal and upstream migration of N. granosa using three experimental treatments: (i) reduced flow (RF) owing to a stream diversion, (ii) natural flow (NF) and (iii) natural flow with artificially increased snail density. For each treatment, snails were differentially tagged and released in a stream without a natural, extant population of N. granosa. 3. Capture rates ranged from 17 to 65% over a 63‐day period following release. Captures on 2–6 days after release measured initial dispersal and migration, whereas longer‐term migration rates were calculated from snails captured 16–63 days after release. Snails under NF displayed positive rheotactic behaviour, with only 3–12% demonstrating initial downstream movement. Under RF, 22–77% of snails moved downstream or showed no bias either way. 4. Initial mean upstream migration rate (UMR) was 0.25, 0.66 and 1.16 m day^?1 under RF, NF and natural flow with increased snail density, respectively. Longer‐term migration rates did not differ significantly between treatments, and the overall mean was 0.62 m day^?1. 5. Principal component analysis and generalised linear models were used to identify habitat characteristics important to UMR, with habitat and reach‐scale hydraulics as the most important factors. The relationship between discharge and UMR suggested it would take 11–35 years for snails to migrate past the most upstream water diversion. However, rates from published studies of neritid snail species migrating en masse or in long lines under natural situations suggested that N. granosa could migrate above stream diversions within 72 days–2.5 years (when in an aggregation) and 29 days–1.1 years (when following in long lines of individuals). 6. An understanding of upstream neritid snail migration can be used for the management and conservation of this and other migratory species in tropical streams.     

Habitat utilization by Hawaiian stream fishes with reference to community structure in oceanic island streams

Synopsis Much of the freshwater biota on high Pacific islands is derived from marine ancestors. Traces of this marine origin are seen in the amphidromous life history patterns of these species. Investigation of the habitat utilization of an assemblage of freshwater gobies in the Hawaiian Islands indicates some resource partitioning within this group. The three major stream species appear to exhibit distinct habitat preferences, which in conjunction with their longitudinal zonation in streams may have served to facilitate their colonization or co-existence.     

Flow velocity underpins microhabitat selection by gobies of the Australian Wet Tropics

1. Water flow is a critical driver of aquatic ecosystem health and function. Amid rising concerns over changing flow regimes, there is an urgent need to understand the functional mechanisms by which flow influences patterns of freshwater biodiversity. 2. We explored the functional link between flow velocity and microhabitat specialisation in a speciose group of freshwater gobies (comprising over half the total fish species richness) within insular streams of the Australian Wet Tropics under base flow conditions. We addressed two particular questions: (i) What is the relative selectivity of species towards streambed composition and water flow velocity? and (ii) Can patterns of microhabitat occupation be explained by differences in intrinsic flow performance among species? To answer these questions, we combined visual field observations of microhabitat use with flow tank assessments of flow speed performance. 3. Tropical freshwater gobies displayed strong specificity towards flow velocities, while being relatively non‐selective towards streambed composition. At opposite extremes of the spectrum, we found Sicyopterus lagocephalus occupying high‐flow (>1.0 ms^?1) microhabitats while Redigobius bikolanus selected slower‐flow (<0.05 ms^?1) areas. These patterns of microhabitat flow specificity were largely explained by the different abilities of species to swim and/or cling to the substratum under these different flow settings. 4. Our findings confirm suggestions that predictable base flows in tropical streams support habitat specialists, which include one species capable of occupying areas of extremely high flow that very few other fishes can withstand. 5. The functional link between flow and gobioid fish distribution patterns could occur throughout tropical streams of the Indo‐Pacific and Caribbean as a widespread phenomenon that may help inform stream flow management guidelines to maintain this substantial component of tropical freshwater biodiversity around the globe.     

Maintaining Biodiversity in Freshwater Ecosystems on Oceanic Islands of the Tropical Pacific

Duringthepastfiveyears,aresearchgroupfromtheHawai'iDivisionofAquaticRe-sourcesandtheLouisianaStaeUniversityMuseumofNaturalSciencehascollaboratedonaseriesofstudiesconcerningthebiologyandconservaionofstreamanimalsintheHawaiianIs-lands.Fromtheviewpointsofbothgeologyandbiology,theseislandsrepresentthenorth-ernmostextensonofPolynesia.StreamanimalsinHawai'ihaverelativesattheleveloffami-ly,genus,andoccasionallyevenspeciesnotonlyinPolynesiabutthroughoutMelanesiaandMicronesiaaswell.Forthisreason…     

Hawaiian biogeography and the islands' freshwater fish fauna

Aim This paper describes known patterns in the distributions and relationships of Hawaiian freshwater fishes, and compares these patterns with those exhibited by Hawaii's terrestrial biota. Location The study is based in Hawaii, and seeks patterns across the tropical and subtropical Indo‐west Pacific. Methods The study is based primarily on literature analysis. Results The Hawaiian freshwater fish fauna comprises five species of goby in five different genera (Gobiidae). Four species are Hawaiian endemics, the fifth shared with islands in the western tropical Pacific Ocean. All genera are represented widely across the Indo‐west Pacific. All five species are present on all of the major Hawaiian islands. All five species are amphidromous – their larval and early juvenile life being spent in the sea. Although there has been some local phyletic evolution to produce Hawaiian endemics, there has been no local radiation to produce single‐island endemics across the archipelago. Nor is there evidence for genetic structuring among populations in the various islands. Main conclusions In this regard, the freshwater fish fauna of Hawaii differs from the well‐known patterns of local evolution and radiation in Hawaiian Island terrestrial taxa. Amphidromy probably explains the biogeographical idiosyncrasies of the fish fauna – dispersal through the sea initially brought the fish species to Hawaii, and gene flow among populations, across the archipelago, has hitherto inhibited the evolution of local island endemics, apparently even retarding genetic structuring on individual islands.     

Life History of the Hawaiian Fish Kuhlia sandvicensis as Inferred from Daily Growth Rings of Otoliths

Kuhlia sandvicensis, the aholehole, is a native Hawaiian fish found in both marine and freshwater habitats. In the lower reaches of streams, they are predators on stream fishes, invertebrates, and insects. Aholehole are an important food fish in the Hawaiian Islands and were often used by ancient Hawaiians in traditional ceremonies. Although aholehole are an important part of stream ecosystems and Hawaiian culture, little is known about their life history, specifically, whether a freshwater phase is obligatory. In this study, light microscopy and electron microprobe techniques were used to analyze otolith daily increments. The analysis estimated age of juveniles and provided information regarding salinity of a fish's habitat at specific points in its life. Sr/Ca profiles from otoliths of juvenile and adult fish from fresh and salt water indicated that this species' use of stream habitats is facultative. Unlike Hawaiian freshwater gobies and at least one other member of the Kuhliidae from the Western Pacific, there is no physiological requirement of fresh water at a specific point in the life cycle of K. sandvicensis. Future research will provide a greater understanding as to the importance of streams as nursery habitats for this species. The research is expected to bolster the argument for maintaining the stream-ocean corridor for access by amphidromous gobies and perhaps also for the aholehole.     

Invertébrés Sans Frontières: Large Scales of Connectivity of Selected Freshwater Species among Caribbean Islands

The freshwater fauna (crustaceans, molluscs, fish) of many tropical islands in the Caribbean and Pacific share an amphidromous life‐cycle, meaning their larvae need to develop in saline conditions before returning to freshwater as juveniles. This community dominates the freshwaters of much of the tropics, but is poorly known and at risk from development, in particular dam construction. Amphidromy can theoretically lead to dispersal between different freshwater areas, even to distant oceanic islands, via the sea. The extent and scale of this presumed dispersal, however, is largely unknown in the Caribbean. Recent genetic work in Puerto Rico has shown that many freshwater species have little or no population structure among different river catchments, implying high levels of connectivity within an island, whereas between‐island structure is unknown. We used genetic techniques to infer the geographic scales of population structure of amphidromous invertebrates (a gastropod and a number of crustacean species) between distant parts of the Caribbean, in particular Puerto Rico, Panama and Trinidad. We found virtually no geographic population structure across over 2000 km of open sea for these freshwater species. This implies that they are indeed moving between islands in sea currents as larvae, meaning that continued recruitment requires a continuum of healthy habitat from the freshwater to marine environment. We further discuss the role of amphidromy and suggest its ecological and biogeographic role may be more important than previously presumed.     

Biogeographic conundrum: Why so few stream nerite species (Gastropoda: Neritidae) in Australia?

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Elevated river discharge enhances the immigration of juvenile catadromous and amphidromous fishes into temperate coastal rivers

Anthropogenic alterations to river flow regimes threaten freshwater biodiversity globally, with potentially disproportionate impacts on species that rely on flow cues to trigger critical life history processes, such as migration for diadromous fishes. This study investigates the influence of river discharge on the abundance of juvenile fish moving into rivers by four temperate catadromous or amphidromous species (common galaxias Galaxias maculatus, spotted galaxias Galaxias truttaceus, climbing galaxias Galaxias brevipinnis and the threatened Australian grayling Prototroctes maraena). Fyke netting or fishway trapping was used to catch juvenile fish moving from estuaries into freshwater in five coastal waterways in south-eastern Australia during the spring migratory period. There was a positive relationship between the probability of high catch rates and mean discharge in September. We also found a positive relationship between discharge and the number of recruits captured 22–30 days later in a flow stressed system. In addition, day-of-year had a strong influence on catch rates, with the peak abundance of juveniles for three species most likely to occur midway through the sampling period (spotted galaxias in October, climbing galaxias in late October and Australian grayling in late October and early November). Our study shows that higher magnitudes of river discharge were associated with increased catches of juvenile catadromous and amphidromous fishes. With a limited supply of environmental water, environmental flows used to enhance immigration of these fishes may be best targeted to maintain small amounts of immigration into freshwater populations in waterways or years when discharges are low and stable. When there are natural, large discharge volumes, relatively large numbers of juvenile fish can be expected to enter coastal waterways and during these times environmental flows may not be required to promote immigration.     

Predator-dependent diel migration by <Emphasis Type="Italic">Halocaridina rubra</Emphasis> shrimp (Malacostraca: Atyidae) in Hawaiian anchialine pools

Diel migration is a common predator avoidance mechanism commonly found in temperate water bodies and increasingly in tropical systems. Previous research with only single day and night samples suggested that the endemic shrimp, Halocaridina rubra, may exhibit diel migration in Hawaiian anchialine pools to avoid predation by introduced mosquito fish, Gambusia affinis, and perhaps reverse migration to avoid the predatory invasive Tahitian prawn, Macrobrachium lar. To examine this phenomenon in greater detail, we conducted a diel study of H. rubra relative abundance and size at 2-h intervals in three anchialine pools that varied in predation regime on the Kona-Kohala Coast of Hawai‘i Island. We found two distinct patterns of diel migration. In two pools dominated by visually feeding G. affinis, the abundance of H. rubra present on the pool bottom or swimming in the water column was very low during the day, increased markedly at sunset and remained high until dawn. In contrast, in a pool dominated by the nocturnal predator M. lar, H. rubra density was significantly lower during the night than during the day (i.e., a pattern opposite to that of shrimp in pools containing fish). In addition, we observed that the mean body size of the shrimp populations varied among pools depending upon predator type and abundance, but did not vary between day and night in any pools. Our results are consistent with the hypothesis that H. rubra diel migratory behavior and size distributions are influenced by predation regime and suggest that diel migration may be a flexible strategy for predator avoidance in tropical pools where it may be a significant adaptive response of endemic species to introduced predators.     

Local adaptation despite high gene flow in the waterfall‐climbing Hawaiian goby,Sicyopterus stimpsoni

Environmental heterogeneity can promote the emergence of locally adapted phenotypes among subpopulations of a species, whereas gene flow can result in phenotypic and genotypic homogenization. For organisms like amphidromous fishes that change habitats during their life history, the balance between selection and migration can shift through ontogeny, making the likelihood of local adaptation difficult to predict. In Hawaiian waterfall‐climbing gobies, it has been hypothesized that larval mixing during oceanic dispersal counters local adaptation to contrasting topographic features of streams, like slope gradient, that can select for predator avoidance or climbing ability in juvenile recruits. To test this hypothesis, we used morphological traits and neutral genetic markers to compare phenotypic and genotypic distributions in recruiting juveniles and adult subpopulations of the waterfall‐climbing amphidromous goby, Sicyopterus stimpsoni, from the islands of Hawai'i and Kaua'i. We found that body shape is significantly different between adult subpopulations from streams with contrasting slopes and that trait divergence in recruiting juveniles tracked stream topography more so than morphological measures of adult subpopulation differentiation. Although no evidence of population genetic differentiation was observed among adult subpopulations, we observed low but significant levels of spatially and temporally variable genetic differentiation among juvenile cohorts, which correlated with morphological divergence. Such a pattern of genetic differentiation is consistent with chaotic genetic patchiness arising from variable sources of recruits to different streams. Thus, at least in S. stimpsoni, the combination of variation in settlement cohorts in space and time coupled with strong postsettlement selection on juveniles as they migrate upstream to adult habitats provides the opportunity for morphological adaptation to local stream environments despite high gene flow.     

Insular Freshwater Fish Faunas of Micronesia: Patterns of Species Richness and Similarity

Freshwater fishes are an important but relatively little known component of the highly diverse fish fauna of Micronesia. Localities supporting communities of freshwater fishes include large high islands, with considerable habitat complexity, and smaller low islands, such as atolls and raised coral islands, with limited freshwater habitat. Both types of islands may support species with adult life history styles that are (a) amphidromous and catadromous, (b) euryhaline (often estuarine), or (c) marine species which enter freshwater from time to time. We compared patterns of species richness and similarity between Micronesian localities for amphidromous and catadromous, euryhaline and marine species (ACEM) pooled, and for amphidromous and catadromous species (ACFW). Species richness of both ACEM and ACFW fishes was greatest on larger high islands compared to smaller high and all low islands. Cluster analysis of similarity indices for ACEM species between localities revealed two faunal components: high islands and low islands. High islands were further partitioned into a Caroline Islands cluster and a separate Mariana Islands cluster. Cluster analysis of ACFW species was more complex. One cluster consisted of a low island and a small high island, both in the Carolines chain and with limited freshwater habitat. The second cluster was partitioned into high islands and low islands that reflected influences of both size and geographical location.