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Human activity can put non-game fishes at higher risk of extinction because of inappropriate management action. Eastern sand darter (Ammocrypta pellucida), a small benthic fish classified as threatened across much of its northern range, inhabits increasingly fragmented sandy habitats and, as a non-game fish, may be easily overlooked in conservation efforts. In this study, the authors use genotype data from nine microsatellite loci and cytochrome oxidase I (COI) sequencing data across its northern native range to re-assess genetic structure and to characterize a newly discovered, geographically disjunct population. Previous microsatellite marker analyses had identified seven distinct population genetic clusters across the region sampled; the analysis of this study showed that the newly discovered population (West Lake, Ontario) exhibits a divergent structure. COI haplotype analysis suggests that a single haplotype recolonized the Great Lakes and surrounding water bodies after the Wisconsinan glacial period, and subsequent fluctuation in water levels and habitat fragmentation resulted in divergence of genetic clusters. Although the novel West Lake population has a common ancestral source with other populations in the broader region, its divergent genetic signature merits its consideration as a separate conservation unit. The analyses of this study highlight the potential conservation implications of the discovery of new populations, particularly those of at-risk species, even within a region that has been genetically well characterized.

Movement is a fundamental aspect of fish ecology, and it therefore represents an important trait to monitor for the management and conservation of fish populations. This is especially true for small benthic fish, as they often inhabit part of the catchment where their movement may be restricted by alterations to river connectivity due to human activity. Still, the movement of these small benthic fish remains poorly understood, partly because of their small size and their cryptic nature. This applies to Percilia irwini, an endangered small darter native to the south-central region of Chile. Its habitat has been affected by the presence of large hydroelectric dams and is currently threatened by the construction of several others. In this study, the authors investigated movement patterns of P. irwini from populations inhabiting different parts of the Biobío catchment, with different levels of connectivity due to natural and/or human-induced features. The authors combined chronological clustering with random forest classification to reconstruct lifelong movements from multi-elemental otolith microchemistry transects. The majority of the movements detected occurred in an undisturbed part of the catchment. These were directional upstream movements occurring between capture sites from the lower and the middle reaches of the river, representing a distance of nearly 30 km, a distance much larger than previously thought. Nonetheless, in the part of the catchment where connectivity was affected by human activity, no such movements were identified. This study shows that connectivity alteration could impede naturally occurring movement and further threaten the resilience of populations of P. irwini. Furthermore, the results presented are used to discuss advantages and disadvantages of microchemistry analysis for studying movement of small benthic fish.

Understanding a species' historical and current distribution is critical when making conservation and management decisions. Recent observations in headwater streams of northern Illinois, USA, where no previous records of Iowa Darters Etheostoma exile occurred, revealed the need to re-evaluate its state-wide distribution. We conducted a series of species distribution modelling procedures coupled with targeted field surveys to generate historical and contemporary distribution models. The historical distribution model indicated E. exile distributions were concentrated across extreme northern Illinois. Both contemporary models included results of the historical model as a model variable. Based on the initial contemporary model, 30 potential Iowa Darter sites, 10 each of three groups representing low, medium and high model scores, were sampled during the summer of 2016. Field surveys provided nine new E. exile localities and presence-absence data from field surveys were then applied to revise the contemporary model. The revised contemporary model suggests a decrease in certain areas of its historic Illinois range, as well as areas where it has potentially expanded its range.

To test hypotheses explaining variation in elaborate male colouration across closely related species groups, ancestral-state reconstructions and tests of phylogenetic signal and correlated evolution were used to examine the evolution of male body and fin colouration in a group of sexually dichromatic stream fishes known as darters (Percidae: Etheostomatinae). The presence or absence of red-orange and blue-green male colour traits were scored across six body regions in 99 darter species using a recently estimated amplified fragment length polymorphism (AFLP) phylogeny for comparative analyses. Ancestral-state reconstructions infer the most recent common ancestor of darters to lack red-orange colour and possess blue-green colour on different body regions, suggesting variation between species is due to independent gains of red-orange and losses of blue-green. Colour traits exhibit substantial phylogenetic signal and are highly correlated across body regions. Comparative analyses were repeated using an alternative phylogenetic hypothesis based on one mitochondrial and two nuclear genes, yielding similar results to analyses based on the AFLP phylogeny. Red-orange colouration in darters appears to be derived; whereas, blue-green appears to be ancestral, which suggests that different selection mechanisms may be acting on these two colour classes in darters.

Robust reproductive isolation was found between the rainbow darter Etheostoma caeruleum and the orangethroat darter Etheostoma spectabile, as more offspring were produced when conspecific males and females were crossed as compared with heterospecific crosses. Furthermore, fewer eggs resulted from heterospecific crosses involving sympatric E. spectabile females than those using allopatric E. spectabile females, while a similar pattern was not observed in heterospecific crosses using E. caeruleum females. These results suggest that reinforcement, i.e. selection for pre-zygotic reproductive barriers driven by reduced hybrid fitness, may have contributed to the evolution and maintenance of reproductive barriers between these potentially hybridizing species in sympatry.

This study examined how colour varies across season and sex in the fantail darter Etheostoma flabellare and the banded darter Etheostoma zonale. Etheostoma flabellare has male-only parental care and exhibited slight sexual dimorphism in overall colour, with no discernible effect of season on colour; whereas E. zonale does not have parental care and exhibited substantial sexual dimorphism in colour, but only in the breeding season. Additionally, antipredator behaviour of E. zonale was compared between males that were fully coloured during the breeding season and males that were partially coloured at that time, but the effects of colour and season were not consistent across males.

In this study, female preferences and behavioural isolation were estimated in a pair of allopatric sister species, Etheostoma duryi and Etheostoma flavum. Dichotomous mate preference trials were conducted to determine whether females prefer to associate with conspecific over heterospecific males and free-spawning assays were conducted to determine whether those preferences translated into behavioural isolation. Dichotomous mate choice trials revealed asymmetric female preference, as female E. flavum preferred conspecific males, whereas female E. duryi showed no preference. Free-spawning assays indicated that behavioural isolation remains incomplete between E. duryi and E. flavum (IB = 0·19). In addition to female mating preferences, male behaviour also appeared to influence mating outcomes as male E. flavum consistently courted conspecific females more often in free-spawning assays whereas male E. duryi did not. The data therefore suggest that despite marked divergence in male nuptial colour, divergence in female preferences between these species may not be sufficient to maintain species boundaries upon secondary contact. These results contrast with similar work in a sympatric darter species pair and may be explained by considering the contributions of reinforcement and differences in colour pattern as well as colour value.

To inquire how male size interacts with alloparental behaviour and mating success in the tessellated darter Etheostoma olmstedi, males were given a choice of nests with or without eggs; subsequent nest occupancy, takeovers and egg deposits were monitored. Subordinate males readily occupied available nests with eggs but were often evicted by dominant males, suggesting that males of all sizes compete for the opportunity to provide allopaternal care in this species.

Incongruence between recognized taxonomy and phylogenetic relationships between two species from a diverse clade (Percidae: Etheostomatinae) of stream fishes was found in a mitochondrial (mt) DNA gene tree. Two darters in subgenus Oligocephalus, Etheostoma uniporum current darter and Etheostoma caeruleum rainbow darter were sampled throughout their sympatric distribution in the Ozark Highlands of the central United States. Sequences from cytochrome (cyt) b and the first intron of the nuclear marker S7 were analysed separately using maximum parsimony and Bayesian methods. Cyt b recovered both species as polyphyletic; E. uniporum haplotypes were interspersed within E. caeruleum. However, both species were monophyletic and non-sister taxa based on S7. The cyt b gene tree pattern is caused by introgressive hybridization resulting in the mtDNA replacement of E. uniporum haplotypes by those of E. caeruleum. Some E. uniporum haplotypes are shared with geographically proximate E. caeruleum, and this is consistent with recent hybridization, while other E. uniporum haplotypes indicate historical sorting of introgressed lineages. The mechanism of introgression is likely asymmetric sneaking behaviour by male E. uniporum, a mating tactic observed in related species. MtDNA replacement may have occurred in E. uniporum due to drift fixation in a historically small female effective population. Additional evidence for darter hybridization will likely be discovered in future molecular genetic surveys of the nearly 200 species in eastern North America.

The a priori null hypotheses that responses to donor, orangethroat darter Etheostoma spectabile, johnny darter Etheostoma nigrum, logperch Percina caprodes, slenderhead darter Percina phoxocephala and blackside darter Percina maculata, extracts would be absent in the three test species, orangethroat darter, johnny darter and logperch, was rejected. This was because orangethroat and johnny darter responded with decreased activity to skin extracts derived from conspecifics and from other members of the assemblage. In contrast, logperch were notable by their lack of a detectable response to any species. Similarities and differences in habitat utilization are seen as a possible basis for these responses.

We tested two hypotheses that describe previously untested assumptions about the appropriate stage(s) for use in determining clutch sizes in darters. Three sequential stages of clutch/ovarian development are recognized: mature oocyte/ovary, ripening oocytc/ovary, and ripe oocyte/ovary. Mean ratios of clutch size to female length (relative clutch size, RCS) were significantly smaller and variances of RCS were significantly greater in ripe females when compared to mature and ripening females. Correlation coefficients for the relationship between clutch size and standard length (S.L.) were significantly greater for mature and ripening females than for ripe females. Mean clutch size, adjusted for the S.L. covariate, was significantly greater in mature and ripening females than in ripe females. Thus, we conclude that counts of eggs from ripe females yield fecundity estimates that are lower and more variable than estimates from counts of oocytes from mature and ripening females collected at the same time. Based on this conclusion, we discuss methods for developing fecundity estimates in darters. Our results may be broadly applicable to other taxa of fishes with group-synchronous ovum development.

Dull coloured, non-breeding males of the darter Etheostoma flabellare‘freeze’ in response to simulated predatory threat. As they enter breeding condition, colour changes are minor and antipredator behaviour does not change. Male E. blennioides, which develop bright green colours in the breeding season, also freeze in response to threats in both breeding and non-breeding states. Male E. spectabile change colours dramatically when breeding, developing intense and contrasting orange, blue, yellow and red colours on their sides, fins and throats. In non-breeding colouration, they freeze when threatened, but in breeding colouration they become less likely to freeze and more likely to flee. These results are discussed both with respect to the hypothesis that changing colours may require changing antipredator behaviour, and in terms of the probable degree of crypticity of these species. Details of the duration of immobility of ‘freezing’ fish and the pattern of subsequent recovery of activity level are also presented.