Small Benthic Fish Research Articles

The role of intraspecific competition in the dispersal of an invasive fish

Abstract Ponto‐Caspian gobies are among the most successful fish invaders in inland waters of Europe and in the Great Lakes of North America. Their invasions appear to comprise a combination of passive and active dispersal mechanisms, both natural and human‐mediated. Despite the significance of Ponto‐Caspian gobies as invasive species, there is little information about the mechanisms underpinning their dispersal. They are relatively small benthic fish, with high site affinity. Thus, actively dispersing individuals must express a behavioural motivation to engage in persistent directional movement. Several recent studies have suggested that inter‐individual behavioural differences in boldness, activity level and tendency to explore might underpin dispersal. In addition, because males are highly territorial, intraspecific competition may lead to density‐dependent dispersal of subordinate individuals. To date, studies on this subject have focused mainly on comparisons between individuals from the core of established populations with those at the margins and, thus, address the outcome of dispersal rather than the mechanism itself. We conducted a series of experiments on the racer goby Babka gymnotrachelus to address the question of what behavioural and physiological traits predict the tendency of an individual to perform dispersal behaviour, specifically considering the role of conspecifics in influencing the tendency to disperse. We used an artificial channel to measure dispersal tendency in this species in combination with behavioural trials. Our results showed that fish with a greater tendency to disperse in an experimental channel grew more slowly and were bolder; that is displayed a greater propensity to emerge from a cryptic background onto a white background and performed worse in prey capture trials. As predicted, intraspecific competition played a primary role in the dispersal of the racer goby. Dominant males showed a tendency to monopolise limited shelters with an outcome that subordinates were forced to disperse upstream. The specific growth rate of individuals appeared to be a good indicator of social position. Subordinate males expressed the lowest rates of growth, probably as a result of long‐term deprivation of food in the presence of dominant conspecifics. They were also quicker to explore novel environments, possibly to search for food. Contrary to expectations, subordinate individuals performed relatively poorly in feeding trials when tested individually. Our findings suggest that intraspecific competition in racer goby males is an important mechanism for active dispersal. It can also influence inter‐individual variation in traits such as boldness and tendency to explore novel environments. Similar responses to competitive interactions may have encouraged the invasive expansion of other Ponto‐Caspian gobies following establishment in new environments, as well as other fish species that exhibit territorial behaviour.

Asymmetric introgression between fishes in the Red River basin of Texas is associated with variation in water quality.

When ecologically divergent taxa encounter one another, hybrid zones can form when reproductive isolation is incomplete. The location of such hybrid zones can be influenced by environmental variables, and an ecological context can provide unique insights into the mechanisms by which species diverge and are maintained. Two ecologically differentiated species of small benthic fishes, the endemic and imperiled prairie chub, Macrhybopsis australis, and the shoal chub, Macrhybopsis hyostoma, are locally sympatric within the upper Red River Basin of Texas. We integrated population genomic data and environmental data to investigate species divergence and the maintenance of species boundaries in these two species. We found evidence of advanced‐generation asymmetric hybridization and introgression, with shoal chub alleles introgressing more frequently into prairie chubs than the reciprocal. Using a Bayesian Genomic Cline framework, patterns of genomic introgression were revealed to be quite heterogeneous, yet shoal chub alleles were found to have likely selectively introgressed across species boundaries significantly more often than prairie chub alleles, potentially explaining some of the observed asymmetry in hybridization. These patterns were remarkably consistent across two sampled geographic regions of hybridization. Several environmental variables were found to significantly predict individual admixture, suggesting ecological isolation might maintain species boundaries.

Feeding Interactions between Fishes in a Coastal Ecosystem in the Southern Gulf of St. Lawrence, Atlantic Canada

AbstractDiet composition, diet similarity, and major predator–prey interactions between the 15 most abundant demersal and pelagic fishes (N = 12,163 stomachs) during the period 1999–2003 were described for Northumberland Strait, a semi‐enclosed, marine coastal ecosystem. Of the five pelagic species, Rainbow Smelt Osmerus mordax ate benthic prey (shrimps and polychaetes); Atlantic Herring Clupea harengus and American Shad Alosa sapidissima consumed small copepods and crab zoeae; and Atlantic Mackerel Scomber scombrus and Alewives Alosa pseudoharengus ate small copepods and moderate amounts of benthic prey and small fishes. Three demersal species were strongly piscivorous but with minimal diet overlap: Sea Ravens Hemitripterus americanus consumed small benthic fishes (especially flounders); Winter Skate Leucoraja ocellata consumed sand lances Ammodytes sp. and Rainbow Smelt; and White Hake Urophycis tenuis consumed Atlantic Herring and Atlantic Mackerel. Cluster analysis revealed seven feeding guilds, of which four contained a single species; consequently, the potential for compensation of predatory function was nonexistent for these guilds. The three multispecies guilds comprised consumers of (1) small copepods and crab zoeae; (2) small benthic invertebrates and polychaetes; and (3) shrimps and crabs. Based on diets, the principal route of energy transfer seemed to be the benthic–detrital pathway; however, confirmation requires representative abundance estimates for the various fishes. Moreover, abundances of many fish species have changed (i.e., most have decreased) since this study was completed, with unknown consequences for food web structure and functioning.

Morphology, sex ratio, and diet of Bothus podas (Delaroche, 1809) in Heraklion Bay, Crete (Greece)

The aim of the present study is to provide morphometric relationships for the Mediterranean wide-eyed flounder Bothus podas (Delaroche, 1809), to identify possible sexual dimorphism, and to determine the feeding habits and the functional role of the species in the ecosystem. Sampling was carried out during a trawl survey off the coast of northern Crete in early May of 2015. A total of 425 specimens were examined (175 males and 250 females; total length (TL) range: 6.2-13.5 cm and 6.0-15.1 cm, respectively). The relationships between TL and weight, interorbital space (IO), body height (BH), and mouth dimensions (horizontal mouth opening: HMO, vertical: VMO mouth opening, and mouth area: MA) were all significant (P < 0.05) and of power types. The estimated value of the exponent b of the length-weight relationship (LWR) for sexes combined was 2.923. Sexual dimorphism was exhibited for the relationships between TL and IO, BH, VMO, HMO, and MA, whereas this was not true for the LWR. The diet spectrum of B. podas includes mainly benthic invertebrates and to a lesser extent small benthic fishes. On one occasion, an individual larva of Fistularia commersonii was recorded in the stomach contents of a male, and this is the first report of any predator of this Lessepsian invader in the Mediterranean.

First report of the parasitic copepod Lernaea cyprinacea (Copepoda: Lernaeidae) on gobioid fishes (Teleostei: Gobonellidae) in southern Europe

Lernaea cyprinacea is a non host-specific parasitic copepod known to infest many freshwater fish species. Outbreaks of infestations by this ectoparasite may cause mass mortality of parasitized fishes. L. cyprinacea has been found mostly on pelagic species. Records on small benthic fish species are less common. Especially rare are infestations of Gobioidei adapted to a benthic life style, with reports restricted to Asia and, in Europe, to the Ponto-Caspian region. Although it is cosmopolitan, L. cyprinacea has rarely been found in Italy. One of the few Italian localities with documented infestations is Lake Trasimeno, a lake with an economically important fishery. Although endoparasites of commercially interesting fish species in this lake are well documented, information about ectoparasites is rare. In May 2015, specimens of two gobioids − Knipowitschia panizzae and Pomatoschistus canestrinii − infested with L. cyprinacea were sampled at the south shore of Lake Trasimeno. Both gobies are not native to the lake. This is the first documentation of gobioid fishes as hosts of L. cyprinacea in Italy and in Europe (outside of the Ponto-Caspian region). Although both gobies are not optimal hosts (small size, short life expectancy) they have the potential to carry and to transmit the parasite in freshwater habitats, e.g. by unintentional introduction with fry of other fish species.

Anatomical Analysis of the Retinal Specializations to a Crypto-Benthic, Micro-Predatory Lifestyle in the Mediterranean Triplefin Blenny Tripterygion delaisi.

The environment and lifestyle of a species are known to exert selective pressure on the visual system, often demonstrating a tight link between visual morphology and ecology. Many studies have predicted the visual requirements of a species by examining the anatomical features of the eye. However, among the vast number of studies on visual specializations in aquatic animals, only a few have focused on small benthic fishes that occupy a heterogeneous and spatially complex visual environment. This study investigates the general retinal anatomy including the topography of both the photoreceptor and ganglion cell populations and estimates the spatial resolving power (SRP) of the eye of the Mediterranean triplefin Tripterygion delaisi. Retinal wholemounts were prepared to systematically and quantitatively analyze photoreceptor and retinal ganglion cell (RGC) densities using design-based stereology. To further examine the retinal structure, we also used magnetic resonance imaging (MRI) and histological examination of retinal cross sections. Observations of the triplefin’s eyes revealed them to be highly mobile, allowing them to view the surroundings without body movements. A rostral aphakic gap and the elliptical shape of the eye extend its visual field rostrally and allow for a rostro-caudal accommodatory axis, enabling this species to focus on prey at close range. Single and twin cones dominate the retina and are consistently arranged in one of two regular patterns, which may enhance motion detection and color vision. The retina features a prominent, dorso-temporal, convexiclivate fovea with an average density of 104,400 double and 30,800 single cones per mm2, and 81,000 RGCs per mm2. Based on photoreceptor spacing, SRP was calculated to be between 6.7 and 9.0 cycles per degree. Location and resolving power of the fovea would benefit the detection and identification of small prey in the lower frontal region of the visual field.

Regulation of pregnane-X-receptor and microRNAs on detoxification-related genes expressions in Mugilogobius abei under the exposure to diclofenac.

Diclofenac (DCF) has been recognized as an emerging contaminant in aquatic environments. Though many studies have investigated the toxic effects of DCF in human and mammals, limited information is available for the responses of genes associated with detoxification metabolisms in non-target aquatic organisms such as fish. In the present study, a small benthic fish Mugilogobius abei, was chosen as the test organism and the effects of DCF on detoxification-related genes at transcriptional level in M.abei were investigated. Partial cDNAs of pregnane-X-receptor (pxr), cytochrome P450 3A (cyp 3a) and alpha-gst were cloned firstly. The responses of cyp 1a, cyp 3a, alpha-gst and p-gp genes and associated microRNAs expressions were measured under different concentrations of DCF exposure (0.5, 5, 50, 500μg/L) for 24h and 168h. Induction of cyp 1a, cyp 3a, alpha-gst, p-gp and pxr mRNA expressions was observed under DCF exposure for different time. Positive concentration-response relationships between DCF concentrations and cyp 1a as well as alpha-gst mRNA expression were observed under DCF exposure for 24h. The similar trend between pxr mRNA expression and cyp 3a gene expression suggested the role of pxr in regulation of its downstream detoxification genes involved in DCF detoxification in M.abei. The negative correlation between miR-27a and p-gp expression under DCF exposure for 24h indicated the role of miRNA in post transcriptional regulation on detoxification-related genes mRNAs in M.abei exposed to DCF. Overall, DCF exposure, even at environmental levels, may interrupt the responses of the detoxification genes in M.abei, which may affect the response of the exposed organism to other pollutants. This work provides implications on the bio-monitoring and risk assessment of DCF in aquatic ecosystems by using of local native fish species.

Feeding Interactions between Fishes in a Coastal Ecosystem in the Southern Gulf of St. Lawrence, Atlantic Canada

AbstractFor the period 1999–2003, diet composition, diet similarity, and major predator–prey interactions between the 15 most abundant demersal and pelagic fishes (N = 12,163 stomachs) were described for Northumberland Strait - a semienclosed, marine, coastal ecosystem. Of the five pelagic species, Rainbow Smelt Osmerus mordax ate benthic prey (shrimps and polychaetes), Atlantic Herring Clupea harengus and American Shad Alosa sapidissima consumed small copepods and crab zoea, and Atlantic Mackerel Scomber scombrus and Alewife A. pseudoharengus ate small copepods and moderate amounts of benthic prey and small fishes. Three demersal species were strongly piscivorous but with minimal diet overlap: Sea Raven Hemitripterus americanus consumed small benthic fishes (especially flounders); Winter Skate Leucoraja ocellatus consumed sand lance (Ammodytes sp.) and Rainbow Smelt; and White Hake Urophycis tenuis consumed Atlantic Herring and Atlantic Mackerel. Cluster analysis revealed seven feeding guilds of which fo...

Effective Dispersal of Caribbean Reef Fish is Smaller than Current Spacing Among Marine Protected Areas

The oceans are deteriorating at a fast pace. Conservation measures, such as Marine Protected Areas, are being implemented to relieve areas from local stressors and allow populations to restore to natural levels. Successful networks of MPAs operate if the space among MPAs is smaller than the dispersal capacity of the species under protection. We studied connectivity patterns across populations in a series of MPAs in the common yellowhead Jawfish, Opistognathus aurifrons. Using the power of genome-wide variation, we estimated that the maximum effective dispersal is 8.3 km. We found that MPAs exchange migrants likely via intermediate unprotected habitats through stepping stone dispersal. At scales >50 km such connectivity is decreased, particularly across the Mona Passage. The MPA network studied would be unable to maintain connectivity of these small benthic fishes if habitat in between them is extirpated. Our study highlights the power of SNPs to derive effective dispersal distance and the ability of SNPs to make inferences from single individuals. Given that overall reef fish diversity is driven by species with life histories similar to that of the yellowhead jawfish, managers face a challenge to develop strategies that allow connectivity and avoid isolation of populations and their possible extinction.

Marine reserves indirectly affect fine-scale habitat associations, but not overall densities, of small benthic fishes.

Many large, fishery‐targeted predatory species have attained very high relative densities as a direct result of protection by no‐take marine reserves. Indirect effects, via interactions with targeted species, may also occur for species that are not themselves targeted by fishing. In some temperate rocky reef ecosystems, indirect effects have caused profound changes in community structure, notably the restoration of predator–urchin–macroalgae trophic cascades. Yet, indirect effects on small benthic reef fishes remain poorly understood, perhaps because of behavioral associations with complex, refuge‐providing habitats. Few, if any, studies have evaluated any potential effects of marine reserves on habitat associations in small benthic fishes. We surveyed densities of small benthic fishes, including some endemic species of triplefin (Tripterygiidae), along with fine‐scale habitat features in kelp forests on rocky reefs in and around multiple marine reserves in northern New Zealand over 3 years. Bayesian generalized linear mixed models were used to evaluate evidence for (1) main effects of marine reserve protection, (2) associations with habitat gradients, including complexity, and (3) differences in habitat associations inside versus outside reserves. No evidence of overall main effects of marine reserves on species richness or densities of fishes was found. Both richness and densities showed strong associations with gradients in habitat features, particularly habitat complexity. In addition, some species exhibited reserve‐by‐habitat interactions, having different associations with habitat gradients inside versus outside marine reserves. Two species (Ruanoho whero and Forsterygion flavonigrum) showed stronger positive associations with habitat complexity inside reserves. These results are consistent with the presence of a behavioral risk effect, whereby prey fishes are more strongly attracted to habitats that provide refuge from predation in areas where predators are more abundant. This work highlights the importance of habitat structure and the potential for fishing to affect behavioral interactions and the interspecific dynamic attributes of community structure beyond simple predator–prey consumption and archetypal trophic cascades.

Habitat preference of combtooth blennies (Actinopterygii: Perciformes: Blenniidae) in very shallow waters of the Ionian Sea, south-eastern Sicily, Italy

Background. In the Mediterranean Sea, habitat preferences of small benthic fishes of shallow waters have been poorly investigated. Although combtooth blennies are not of interest for fisheries, they are often the dominant fish community on rocky bottoms within the first meter of depth and in tidal pool environments, playing so an important role in the functioning of the coastal ecosystem. This study represents the first detailed investigation concerning habitat preference and depth distribution of combtooth blennies in the Ionian Sea. Materials and methods. Occurrence and abundance of species were investigated by a non-destructive visual census method using snorkelling or SCUBA diving. We investigated 5 habitat types (bathymetric intervals between 0 and 3 m of depth) to find difference in species richness and abundance in each one and between. Studies were conducted from June to October 2014 in the south-eastern coast of Sicily. Statistical method of the electivity index (EI) and principal component analysis (PCA) were utilized for the evaluation of the habitat preference of 11 species of combtooth blennies. Depth distribution for each species was box-plotted. For each habitat and depth, the Shannon–Wiener Index (H’) and the Simpson Index (D) were calculated. Diversity profiles were performed in order to give a better understanding of the correlation between diversity and both habitat types and depth. Results. During a total of 2485 observations of combtooth blennies a total of 11 species were recorded: Aidablennius sphynx (Valenciennes, 1836); Coryphoblennius galerita (Linnaeus, 1758); Lipophrys trigloides (Valenciennes, 1836); Microlipophrys canevae (Vinciguerra, 1880); Parablennius gattorugine (Linnaeus, 1758); Parablennius incognitus (Bath, 1968); Parablennius pilicornis (Cuvier, 1829); Parablennius sanguinolentus (Pallas, 1814); Parablennius zvonimiri (Kolombatović, 1892); Salaria pavo (Risso, 1810); Scartella cristata (Linnaeus, 1758). The maximum Shannon–Wiener Index (H’= 2.335) and Simpson Index (D = 0.898) values were recorded on “rocks with algal cover”; while, concerning the depth, the maximum values of both indices were recorded within the first meter. The diversity indices showed the highest values in the most heterogeneous habitat types. There was a negative correlation between depth and diversity indices. Results showed a clear habitat preference for the following species: A. sphynx, P. gattorugine, P. sanguinolentus, S. pavo and S. cristata. Conclusion. This study highlights the importance of depth and habitat heterogeneity in biological diversity, species richness and population abundances of combtooth blennies in the Mediterranean Sea.

Comparison of Visual Survey and Mark–Recapture Population Estimates of a Benthic Fish in Hawaii

AbstractVisual surveys are conducted to rapidly estimate population densities of stream fishes, often without calibration against more established or more widely used methods to determine precision and accuracy or to correct for potential biases. We compared population density estimates from a visual survey (VS) point quadrat method widely used in Hawaii with estimates from “in hand” individual and batch mark–recapture (BMR) methods. Visual survey sampling and individual mark–recapture (IMR) sampling were conducted in three watersheds that represent gradients of land use and prevalence of nonnative poeciliid fishes on the Island of Hawaii. Focusing on adult O‘opu Nākea Awaous stamineus, VSs were conducted prior to IMR events to allow direct comparisons of results independent of location and time. Density estimates of O‘opu Nākea from VS and IMR samplings were strongly correlated, although VS estimates were generally higher and underrepresented exceptionally large fish. Batch mark–recapture estimates of O‘opu Nākea densities were conducted for comparison with VSs at 13 sites across the archipelago. Estimates of VSs were not significantly different from BMR estimates. Estimates of VSs also exhibited less variance than did BMR estimates across sites. General linear models showed that the relationship between VS and IMR estimates varied significantly among watersheds but not seasons and that land use was associated with a greater mismatch between VS and BMR estimates of population density. These findings indicate that visual surveys using a point quadrat method are an efficient and accurate approach for estimating the abundance of small benthic fishes, such as O‘opu Nākea, in wadeable streams and that obtaining absolute densities or size distributions from VS methods would benefit from a calibration with IMR not BMR estimates.Received July 15, 2015; accepted February 23, 2016 Published online June 29, 2016

Seasonal and age-related migrations in the unexpectedly long-lived benthic fish the lesser weever, Echiichthys vipera (Cuvier, 1829)

A study of an inshore southern North Sea population of lesser weever, Echiichthys vipera, on the Suffolk coast, England, found this small, abundant, benthic fish to reach an age of 15 years and suffer an adult mortality rate of only 0.23 y−1. The maximum length observed of 195 mm Standard length (SL) (225 mm total length, TL) was the greatest yet reported and many individuals &gt;140 mm SL (163 mm TL) were caught between 2009 and 2012. Previous studies have reported a maximum of 160 mm TL and a von Bertalanffy asymptotic TL of 150.3 mm. Age structure analysis showed that recruitment into the local inshore Sizewell population continued until 5 or more years of age. A 6 year age of recruitment corresponds to the age when they have been reported to have disappeared from offshore locations and previously assumed to have died from old age. Regular seasonal changes in local abundance were observed with peak captures during May, presumably caused by seasonal immigration, followed by a summer minimum and a second, more variable, maximum in early autumn before the winter minimum. The winter minimum in captures may be due to either inactivity or offshore migration. Lesser weever has evolved a long-lived, slow growing, life history strategy unusual for small benthic fish in the southern North Sea. By spending long periods hidden in sand, using venom for defence and remaining inactive for an extended period each winter, lesser weever has adopted a strategy which favours high survival and increased longevity.

Movement of three stream‐resident balitoroid loaches and a goby in a Hong Kong hillstream

AbstractWe undertook a mark–recapture study of three pool‐dwelling balitoroid loaches and a nonmigratory goby in a Hong Kong hillstream, which extended over a 35‐week period. The target species were Pseudogastromyzon myersi and Liniparhomaloptera disparis (Balitoridae), Schistura fasciolata (Nemacheilidae) and Rhinogobius duospilus (Gobiidae). Recapture rates of tagged individuals were high (58% overall), especially for the abundant R. duospilus (70% of 412 tagged) and P. myersi (57% of 762 tagged). At the end of the study, most recaptured individuals (78% of gobies, 62% of P. myersi, 42% of L. disparis and 67% of S. fasciolata) were within the pools where they had been released. Of those that had moved elsewhere, maximum displacements ranged from 46 m (R. duospilus) to 101 m (P. myersi), giving rise to a strongly leptokurtic distribution of movement distances. Mean displacements after 35 weeks ranged from only 2.9 m (R. duospilus) to 10.0 m (L. disparis). The sedentary behaviour of these species was more marked than most other small benthic fishes, although our study excluded the breeding period and wet season when high flows might have stimulated movement. Among the mobile P. myersi individuals, most travelled upstream, whereas mobile gobies tended to move downstream. Juveniles of P. myersi and R. duospilus, as well as large adult P. myersi, were especially sedentary, while mobile adult P. myersi that travelled further showed smaller growth increments. The highly sedentary habits of all four fishes suggest that they may exert persistent local top‐down control of benthic communities within pools in Hong Kong streams.

Molecular pathways associated with the intersex condition in rainbow darter (Etheostoma caeruleum) following exposures to municipal wastewater in the Grand River basin, ON, Canada. Part B

Rainbow darter (Etheostoma caeruleum; RBD) is a small benthic fish found in North America. This species is sensitive to sewage effluent, and intersex is found in up to 80% of males in near-field areas in the Grand River, Ontario, Canada. To learn more about the molecular signaling cascades associated with intersex, a developed customized oligonucleotide microarray (4×180K) using next generation sequencing was developed to characterize the transcriptome in the gonad of male and female RBD. Gene expression profiling was performed in males and females from both a reference site and a polluted site. Males with and without intersex condition from the areas closest to effluent outfalls were compared to males and females from a reference site. Microarray analysis revealed that there was increased mRNA abundance for genes associated with oogenesis in intersex males (i.e. the presence of eggs within the testis), and a decrease in mRNA abundance for genes associated with spermatid development. In females exposed to effluent, cell processes related with hatching and ovulation were down-regulated, and genes involved in immune responses were increased in abundance. In the non-intersex males exposed to effluent, cell processes such as sperm cell adhesion were decreased at the transcript level relative to males from the reference site. Microarray analysis revealed that heat shock proteins (HSP) were significantly increased in non-intersex males exposed to effluent; however, HSPs were not differentially expressed in intersex males exposed to the effluent. Genes involved in sex differentiation (sox9, foxl2 and dmrt1) and reproduction (esr1, esrb, ar, vtg, cyp19a1 and cyp11a) were measured in males, females, and intersex individuals. Consistent with the intersex condition, many transcripts showed an intermediate expression level in intersex males when compared to phenotypic males and females. This study improves our knowledge regarding the molecular pathways that underlie the intersex condition and develops a suite of qPCR bioassays in RBD that are able to discriminate pollutant-exposed males without intersex from those males with intersex. Part A of this study reports on the effects of municipal wastewater effluents (MWWEs) on RBD in the Grand River and demonstrates that there are disruptions in higher level endpoints that include altered steroid levels. Here we develop a new tool for assessing and monitoring the intersex condition in RBD in polluted natural environments and begin to characterize gene networks that are associated with the condition.

Whole organism responses and intersex severity in rainbow darter (Etheostoma caeruleum) following exposures to municipal wastewater in the Grand River basin, ON, Canada. Part A

Municipal wastewater effluents (MWWEs) contain anthropogenic substances that can exhibit endocrine-disrupting activity. These complex mixtures have been observed to exert adverse effects on fish. Rainbow darter (Etheostoma caeruleum, RBD) is a small benthic fish that is widespread throughout the Grand River, Ontario, Canada, and has been previously shown to be adversely affected by MWWE exposure in this watershed. The objectives of this study were to quantify biological responses in this sentinel species and intersex severity in male fish, in relation to the area of urbanization. It focused on RBD populations adjacent to wastewater outfalls in the Grand River watershed. In May 2011, nine sites across the urban gradient were selected to evaluate the impact of MWWEs. Endpoints for energy storage (i.e. condition factor, k; liversomatic index, LSI) as well as reproductive endpoints (i.e. gonadosomatic index, GSI; gonad development, hormone production), and intersex were assessed in the fish. Rainbow darter showed a high incidence of intersex downstream of the wastewater outfalls, especially below the largest treatment plant outfall at Kitchener (∼85%). We applied an intersex index (score from 0 to 7) that considers the number of eggs within the testis and the stage of maturation of the egg. RBD exposed downstream of the largest wastewater outfall at Kitchener had a score of 3.81±0.37 compared to upstream to the urban areas where there were no intersex males found other than a single individual with a score 1 (average intersex score of site 0.06±0.06). In addition, several fish associated with the Kitchener outfall had macroscopic vitellogenic eggs in the testes (intersex scores 5 and 6). The sub-population of fish located at the wastewater outfall also showed a tendency towards skewed sex ratios (greater proportion of females to male fish) compared to the population at the reference sites. Male fish inhabiting the urbanized area of the Grand River also showed reduced levels of testosterone (T) and 11-ketotestosterone (11KT). Intersex males had the lower levels of 11KT relative to the upstream reference fish but could not be distinguished from normal males collected at the exposed sites. Despite the high levels of intersex at these sites, no relationships were evident among intersex severity and other measured endpoints such as GSI, LSI or in vitro steroid production. The effects observed appear to be associated with urbanization and exposure to treated MWWEs in the watershed. Although intersex incidence and severity was a very good indicator of wastewater exposure, intersex could not be directly linked to other effects in this wild population. The effects of MWWEs on transcriptional changes in adult RBD exposed to the effluents are reported in the corresponding report (Part B).

Extensive dispersal of Roanoke logperch (Percina rex) inferred from genetic marker data

AbstractThe dispersal ecology of most stream fishes is poorly characterised, complicating conservation efforts for these species. We used microsatellite DNA marker data to characterise dispersal patterns and effective population size (Ne) for a population of Roanoke logperch Percina rex, an endangered darter (Percidae). Juveniles and candidate parents were sampled for 2 years at sites throughout the Roanoke River watershed. Dispersal was inferred via genetic assignment tests (ATs), pedigree reconstruction (PR) and estimation of lifetime dispersal distance under a genetic isolation‐by‐distance model. Estimates of Ne varied from 105 to 1218 individuals, depending on the estimation method. Based on PR, polygamy was frequent in parents of both sexes, with individuals spawning with an average of 2.4 mates. The sample contained 61 half‐sibling pairs, but only one parent–offspring pair and no full‐sib pairs, which limited our ability to discriminate natal dispersal of juveniles from breeding dispersal of their parents between spawning events. Nonetheless, all methods indicated extensive dispersal. The AT indicated unrestricted dispersal among sites ≤15 km apart, while siblings inferred by the PR were captured an average of 14 km and up to 55 km apart. Model‐based estimates of median lifetime dispersal distance (6–24 km, depending on assumptions) bracketed AT and PR estimates, indicating that widely dispersed individuals do, on average, contribute to gene flow. Extensive dispersal of P. rex suggests that darters and other small benthic stream fishes may be unexpectedly mobile. Monitoring and management activities for such populations should encompass entire watersheds to fully capture population dynamics.

Molecular signatures in rainbow darter (Etheostoma caeruleum) inhabiting an urbanized river reach receiving wastewater effluents

Rainbow darter (Etheostoma caeruleum) is a small benthic fish species found in North America that are abundant and distributed throughout the Grand River watershed, ON, Canada. Rainbow darter exhibit intersex in males at sites adjacent to municipal wastewater effluents (MWWE). In October 2010, female and male rainbow darter were collected at 3 sites (1 upstream reference and 2 downstream exposed sites) in the Grand River near the cities of Kitchener and Waterloo. The primary objectives of this research were (1) to characterize the responses of whole organism endpoints (i.e. condition factor (K), liversomatic (LSI) and gonadosomatic index (GSI), histopathology) to MWWEs and (2) to identify transcripts showing altered steady state abundance with exposure to MWWE in fish inhabiting municipal wastewater effluent-exposed areas. Genes measured in this study included vitellogenin, Sry-box containing protein 9 (sox9), forkhead box L2 (foxl2), doublesex and mab-3 related transcription factor 1 (dmrt1), cytochrome P450, family 11, subfamily A, polypeptide 1 (cyp11a) as well as estrogen (esr1, esrb) and androgen (ar) receptors. There were no changes in condition factor; however, there was a significant increase in LSI and a decrease in GSI in fish inhabiting downstream environments when compared with fish collected from the reference site. Males had a high incidence (∼70%) of intersex in downstream sites; characterized by the presence of oocytes within the testis. In the gonad, there were sex specific differences for genes related to sexual differentiation; dmrt1 was only expressed in males whereas foxl2 and sox9 were highly expressed in females compared to males. Expression levels of ar and esr1 were higher in females than males. Conversely, esrb was not differentially expressed between sexes or among sites. There were no differences detected for the genes investigated within sex among sites. This study is the first to report on gene expression changes in the rainbow darter, with emphasis on the differences in transcript abundance between sexes and how these changes relate to exposures to MWWEs. Molecular approaches are being investigated for their potential application to field ecotoxicology, and molecular bioassays for relevant, sentinel species in environmental monitoring programs are required to better understand the impact of anthropogenic impacts on species at risk in river systems.

Trophic transfer and accumulation of mercury in ray species in coastal waters affected by historic mercury mining (Gulf of Trieste, northern Adriatic Sea)

Total mercury (Hg) and monomethylmercury (MMHg) were analysed in the gills, liver and muscle of four cartilaginous fish species (top predators), namely, the eagle ray (Myliobatis aquila), the bull ray (Pteromylaeus bovinus), the pelagic stingray (Dasyatis violacea) and the common stingray (Dasyatis pastinaca), collected in the Gulf of Trieste, one of the most Hg-polluted areas in the Mediterranean and worldwide due to past mining activity in Idrija (West Slovenia). The highest Hg and MMHg concentrations expressed on a dry weight (d.w.) basis were found in the muscle of the pelagic stingray (mean, 2.529 mg/kg; range, 1.179-4.398 mg/kg, d.w.), followed by the bull ray (mean, 1.582 mg/kg; range, 0.129-3.050 mg/kg d.w.) and the eagle ray (mean, 0.222 mg/kg; range, 0.070-0.467 mg/kg, d.w.). Only one specimen of the common stingray was analysed, with a mean value in the muscle of 1.596 mg/kg, d.w. Hg and MMHg contents in the bull ray were found to be positively correlated with species length and weight. The highest MMHg accumulation was found in muscle tissue. Hg and MMHg were also found in two embryos of a bull ray, indicating Hg transfer from the mother during pregnancy. The number of specimens and the size coverage of the bull rays allowed an assessment of Hg accumulation with age. It was shown that in bigger bull ray specimens, the high uptake of inorganic Hg in the liver and the slower MMHg increase in the muscle were most probably due to the demethylation of MMHg in the liver. The highest Hg and MMHg contents in all organs were found in the pelagic stingray, which first appeared in the northern Adriatic in 1999. High Hg and MMHg concentrations were also found in prey species such as the banded murex (Hexaplex trunculus), the principal prey of the eagle rays and bull rays, the anchovy (Engraulis encrasicholus) and the red bandfish (Cepola rubescens), which are preyed upon by the pelagic stingray, as well as in zooplankton and seawater. Based on previously published data, a tentative estimation of MMHg bioamagnification was established. The average increase in MMHg between seawater, including phytoplankton, and zooplankton in the Gulf was about 10(4), and MMHg in anchovy was about 50-fold higher than in zooplankton. The bioaccumulation of MMHg between seawater and small pelagic fish (anchovy) amounted to 10(6) and between water and the muscle of larger pelagic fish (pelagic stingray) to 10(7). The MMHg increase between surface sediment and benthic invertebrates (murex) and between benthic invertebrates and small benthic fish was 10(2). Ultimately, the trophic transfer resulted in a 10(3) accumulation of MMHg between water and muscle of larger benthic fish (bull ray, eagle ray, common stingray), suggesting lower bioaccumulation by benthic feeding species.

Movement patterns of small benthic fish in lowland headwater streams

Summary Understanding the mechanism governing the movements of stream‐dwelling fish is a fundamental issue associated with a diverse range of ecological processes. We examined the movement pattern of a small benthic species, Lefua echigonia, using a mark–recapture technique in small headwater streams. Our main objectives were as follows: (i) to assess the movement range and direction of L. echigonia; (ii) to examine how biotic and abiotic factors affect emigration and immigration patterns and (iii) to assess movement patterns at the stream scale. Stationary fish accounted for the greatest proportion of recaptured adult L. echigonia. Seventy‐five per cent of recaptured individuals moved &lt;100 m in 12 months after release, and the maximum movement distance was 500 m upstream. Moreover, adult L. echigonia showed significant upstream bias in movements. According to the results of the generalised linear mixed model GLMM analysis, movement of adult L. echigonia was associated mainly with the density of conspecifics, rather than with habitat quality and body size. Adult L. echigonia were mainly stationary in relatively high‐density sections, and emigration tended to increase with decreasing density. Moreover, mobile individuals were prone to move into relatively high‐density sections. The proportion of mobile fish showed a strong positive correlation with catchment area at the stream scale. This result indicates that stream discharge can influence the mobility of adult L. echigonia. During baseflow, the water was static in a number of stream sections, which suggests that the influence of catchment area on mobility may be more a function of freshets than normal water flow.