Gymnotiform Research Articles

Gymnotiform electric fishes of the Tres Fronteras region of the western Amazon

Gymnotiform electric knifefishes are an important yet undersampled component of the Neotropical aquatic biota. We report on the gymnotiform fauna of the Tres Fronteras region located at the triple border of Brazil, Colombia, and Peru in the biodiverse western Amazon. The presence of at least 33 species of gymnotiforms in the Tres Fronteras region is validated from recent sampling efforts and the review of previously collected materials. A key is provided for the identification of the species that have been collected from the region. We comment on the diversity of habitat utilization and intraspecific colour variation of some species.

Exploring potential drivers of brain size variation in the electric fish Brachyhypopomus occidentalis

Characterizing the factors that shape variation in brain size in natural populations is crucial to understanding the evolution of brain size in animals. Here, we explore how relative brain size and brain allometry vary with drainage, predation risk and sex in natural populations of the electric knifefish Brachyhypopomus occidentalis. Fish were sampled from high and low predation risk sites within two independent river drainages in eastern and central Panamá. Overall, we observed low variation in brain-body size allometric slopes associated with drainage, predation risk and sex category. However, we observed significant differences in allometric intercepts between predation risk sites. We also found significant differences in relative brain mass associated with drainage, as well as significant differences in absolute brain mass associated with drainage, predation risk and sex category. Our results suggest potential constraints in brain-body allometry across populations of B. occidentalis. However, both drainage and predation risk may be playing a role in brain mass variation among populations. We suggest that variation in brain mass in electric fishes is affected by multiple extrinsic and intrinsic factors, including geography, environmental complexity, social interaction and developmental or functional constraints.

Electrocommunication signals and aggressive behavior vary among male morphs in an apteronotid fish, Compsaraia samueli.

Within-species variation in male morphology is common among vertebrates and is often characterized by dramatic differences in behavior and hormonal profiles. Males with divergent morphs also often use communication signals in a status-dependent way. Weakly electric knifefish are an excellent system for studying variation in male morphology and communication and its hormonal control. Knifefish transiently modulate the frequency of their electric organ discharge (EOD) during social encounters to produce chirps and rises. In the knifefish Compsaraia samueli, males vary extensively in jaw length. EODs and their modulations (chirps and rises) have never been investigated in this species, so it is unclear whether jaw length is related to the function of these signals. We used three behavioral assays to analyze EOD modulations in male C. samueli: (1) artificial playbacks, (2) relatively brief, live agonistic dyadic encounters, and (3) long-term overnight recordings. We also measured circulating levels of two androgens, 11-ketotestosterone and testosterone. Chirp structure varied within and across individuals in response to artificial playback, but was unrelated to jaw length. Males with longer jaws were more often dominant in dyadic interactions. Chirps and rises were correlated with and preceded attacks regardless of status, suggesting these signals function in aggression. In longer-term interactions, chirp rate declined after 1week of pairing, but was unrelated to male morphology. Levels of circulating androgens were low and not predictive of jaw length or EOD signal parameters. These results suggest that communication signals and variation in male morphology are linked to outcomes of non-breeding agonistic contests.

A new taxonomist-curated reference library of DNA barcodes for Neotropical electric fish (Teleostei: Gymnotiformes)

AbstractDNA barcoding is a useful tool for identifying species; however, successful barcode-based identification requires a reference library of barcode sequences from accurately identified specimens. Here we present a reference library of COI barcode sequences for the Neotropical electric knifefish, order Gymnotiformes (Teleostei: Ostariophysi), a model taxon for studies of tropical diversification and biogeography, genomics, behaviour and neurobiology. Our library contains barcodes for 167 of the c. 270 valid species of gymnotiforms derived from geo-referenced museum voucher specimens, and includes sequences from 26 type specimens and 21 specimens from type localities, most of which we collected. To assess the state of gymnotiform barcodes in two main public barcode repositories, GenBank and BOLD, we compared the barcodes in these databases to our reference library. Our analysis shows that a considerable proportion of gymnotiform barcodes in GenBank and BOLD are mis- or unidentified. We encourage taxonomists to develop and publish barcode reference libraries composed of carefully curated barcode sequences.

Mosaic Evolution of Craniofacial Morphologies in Ghost Electric Fishes (Gymnotiformes: Apteronotidae)

Ghost electric knifefishes (Gymnotiformes: Apteronotidae) are a dominant component of the species diversity and biomass of large lowland rivers in Greater Amazonia, including 77 species that exhibit diverse craniofacial morphologies associated with trophic and secondary sexual traits. Here we use open-source computed micro-tomography (µCT) scans to generate 3D reconstructions for a majority of apteronotid species and almost all valid genera, and geometric morphometric and phylogenetic analyses to explore patterns of skull evolution. As with most vertebrates, principal component 1 (PC1) summarizes variance from brachycephalic to dolichocephalic morphologies, previously described as heterocephaly, and PC2 summarizes variance from recurved (upturned) to decurved (downturned) snout morphologies, described here as heterorhynchy. Phylomorphospace and traitogram analyses found instances of both convergent and divergent evolution along both of the first two PC axes, as well as a preponderance of clades characterized by morphological stasis or phylogenetic conservatism. Certain phenotypic combinations predominate among species and clades, including elongated-downturned snouts and foreshortened-upturned snouts, while other phenotypic combinations are not observed, including elongated-upturned snouts and foreshortened-downturned snouts. These results highlight the power of 3D geometric morphometrics in the study of craniofacial evolution and indicate developmental or functional constraints in the evolution of craniofacial phenotypes in an ecologically dominant clade of riverine Amazonian fishes.

Reproductive effort and terminal investment in a multispecies assemblage of Amazon electric fish

AbstractThe terminal investment hypothesis (TIH) predicts that individuals with favorable prospects for future reproduction (i.e., high residual reproductive value, RRV) should moderate current reproductive investment in favor of growth, survival, and future reproduction, whereas those with low RRV should “terminally invest” by diverting somatic resources towards current reproduction at the expense of future reproduction. However, support for the TIH in wild animal populations is fragmentary, and the ecological contexts of terminal investment remain poorly known. We report a remarkable case of simultaneous terminal investment involving five sympatric species of the electric knifefish genus Brachyhypopomus, from Amazonian floodplain and terra firme stream habitats. We found that terminal investment is synchronized by seasonal breeding, in response to circannual environmental variation in mortality risk. Four species exhibit a uniseasonal iteroparous (annual) life history with complete post‐reproductive mortality after a single breeding season. One species (Brachyhypopomus beebei) exhibits a 2‐year multiseasonal iteroparous life history with breeding in two seasons and post‐reproductive mortality after the second. In mature females and (most) males of the annual species, as well as in both mature female and male second‐year (but not first‐year) B. beebei, we documented an increase in two metrics of reproductive effort (size‐adjusted gonad mass and electric signal amplitude) and a concomitant reduction in somatic condition (size‐adjusted somatic mass), all in response to proximity to the end of the common breeding season, when RRV approximates zero. In mature first‐year B. beebei, we documented neither an increase in reproductive effort nor a decline in somatic condition, implying an alternative strategy of reproductive restraint. Our findings support Kirkwood's disposable soma theory, which posits that death by reproductive exhaustion can be delayed if terminal investment is replaced by reproductive restraint, allowing individuals to survive and breed in a subsequent season. Deferral of the terminal investment response in annual species, and the origin of a gonadal regression‐regeneration sequence, may open pathways for rapid evolutionary transitions to multiseasonal iteroparity. Excepting the age (year‐group) dependency of terminal investment in B. beebei, we were unable to identify intrinsic cues or extrinsic environmental cues for the terminal investment response in Brachyhypopomus.

The weakly electric fish, Apteronotus albifrons, actively avoids experimentally induced hypoxia

Anthropogenic environmental degradation has led to an increase in the frequency and prevalence of aquatic hypoxia (low dissolved oxygen concentration, DO), which may affect habitat quality for water-breathing fishes. The weakly electric black ghost knifefish, Apteronotus albifrons, is typically found in well-oxygenated freshwater habitats in South America. Using a shuttle-box design, we exposed juvenile A. albifrons to a stepwise decline in DO from normoxia (> 95% air saturation) to extreme hypoxia (10% air saturation) in one compartment and chronic normoxia in the other. On average, A. albifrons actively avoided the hypoxic compartment below 22% air saturation. Hypoxia avoidance was correlated with upregulated swimming activity. Following avoidance, fish regularly ventured back briefly into deep hypoxia. Hypoxia did not affect the frequency of their electric organ discharges. Our results show that A. albifrons is able to sense hypoxia at non-lethal levels and uses active avoidance to mitigate its adverse effects.

Myxobolus freitasi n. sp. (Myxozoa: Bivalvulida), a parasite of the brain of the electric knifefish in the Brazilian Amazon region.

A total of 30 specimens of the Amazonian electric knifefish, Brachyhypopomus beebei Schultz, 1944 (Gymnotiformes: Hypopomidae), were collected from the Peixe-Boi River in the state of Pará, Brazil (1°06'59" S; 47°18'26" W). Fragments of the brain tissue were extracted for analysis via optical microscopy, and 18 specimens (60%) presented microparasites of the genus Myxobolus, with unequal capsules. The spores were 18.6 µm (17.7-19.8 µm) long and 8.6 µm (8.4-9.0 µm) wide; the largest polar capsule was 13.0 µm (12.4-13.4 µm) long and 5.6 µm (5.3-6.0 µm) wide, and the smallest capsule was 5.0 µm (4.5-5.3 µm) long and 2.5 µm (2.3-2.6 µm) wide. Infected brain fragments were extracted for histological processing and staining with hematoxylin-eosin and Ziehl-Neelsen. Some fragments were conserved in ethanol for molecular genetics analysis. A partial sequence of the 18S DNA gene was obtained from the spores, which did not correspond to any other sequences deposited in GenBank, although it did form a clade with other Myxobolus parasites of the nervous system. The morphological data, together with molecular phylogeny, supported the designation of a new species Myxobolus freitasi n. sp.

Acute toxicity of a deltamethrin based pesticide (DBP) to the Neotropical electric fish Microsternarchus cf. bilineatus (Gymnotiformes)

ABSTRACT Deltamethrin is a pyrethroid insecticide widely used in pest control in Brazilian agriculture. The intensive and disordered use of this pesticide in the Amazon region can drive it into aquatic ecosystems in several ways, but mainly by runoff and leaching. The present study was conceived to determine the acute toxicity (LC50) of a deltamethrin based pesticide (DBP) and to characterize its effects on two biochemical biomarkers, glutathione-S-transferase (GST) and acetylcholinesterase (AChE), in tissues of the electric fish Microsternarchus cf. bilineatus. Fishes were exposed to concentrations of 1, 2, 3, 4 and 5 μg L-1 of DBP for up to 96 hours. For each treatment, the absolute activity of the enzymes GST (muscle and liver) and AChE (muscle and nervous tissue) were analyzed. The LC50-96 h for Microsternarchus cf. bilineatus was 2.15 μg L-1, the lowest concentration registered for an Amazonian fish species so far. None of the concentrations tested of this insecticide affected AChE activity for the exposure period tested. A significant increase in muscle GST activity was detected only for concentrations of 2 and 3 μg L-1.

Parasitism by Clinostomum marginatum (Clinostomidae) in neotropical electric fish (Gymnotiformes) in the Brazilian Amazon

Gymnotiformes are currently recognized as electric fish, the vast majority of which emit low power electrical discharges. Clinostomum trematodes have been the subject of numerous investigations in several host species due to the ability of metacercariae of this genus to infect the muscles, fins and internal organs of fish. The present study aimed to record, for the first time, the occurrence of parasitism by Clinostomum marginatum in Gymnotiformes from the natural environments of two tributaries of the Amazon River in the Brazilian Amazon. Two specimens of Gymnotus coropinae, ten specimens of Brachyhypopomus brevirostris, two specimens of Gymnorhamphycthys petiti and one specimen of Microsternarchus bilineatus were captured. Infection with C. marginatum was identified in the form of 12 encysted metacercariae in the epidermis, with a prevalence of 0.1%, a mean infection intensity of 12/fish and an average abundance of 1.2. This is the first record of parasitism by the C. marginatum in South American Gymnotiformes of the B. brevirostris species.

Electric eels galore: microsatellite markers for population studies

Abstract Fourteen novel microsatellite loci are described and characterized in two species of electric eels, Electrophorus variiand E. voltaifrom floodplains and rivers of the Amazon rainforest. These loci are polymorphic, highly informative, and have the capacity to detect reliable levels of genetic diversity. Likewise, the high combined probability of paternity exclusion value and low combined probability of genetic identity value obtained demonstrate that the new set of loci displays suitability for paternity studies on electric eels. In addition, the cross-amplification of electric eel species implies that it may also be useful in the study of the closely related E. electricus, and to other Neotropical electric fishes (Gymnotiformes) species as tested herein.

Reproductive life-history strategies in a species-rich assemblage of Amazonian electric fishes.

The reproductive biology of only a small fraction of Neotropical freshwater fishes has been described, and detailed comparative studies of reproductive life-history variation in the Neotropical ichthyofauna are lacking. Here we describe interspecific variation in reproductive life history for a multi-species assemblage of the electric knifefish genus Brachyhypopomus (Hypopomidae: Gymnotiformes: Ostariophysi) from Amazonian floodplain and terra firme stream systems. During a year-round quantitative sampling program, we collected and measured key life-history traits from 3,410 individuals. Based on oocyte size distributions, and on circannual variation in gonadosomatic indices, hepatosomatic indices, and capture-per-unit-effort abundance of reproductive adults, we concluded that all species exhibit a single protracted annual breeding season during which females spawn fractionally. We found small clusters of post-larval individuals in one floodplain species and one terra firme stream species, but no signs of parental care. From analyses of body size-frequency distributions and otolith growth increments, we concluded that five species in our study area have approximately one-year (annual) semelparous life history with a single reproductive period followed by death, while two species have a two-year iteroparous life history, with breeding in both year-groups. Despite predictions from life-history theory we found no salient correlations between life history strategy (semelparity or iteroparity) and habitat occupancy (floodplain or terra firme stream). In the iteroparous species B. beebei, we documented evidence for reproductive restraint in the first breeding season relative to the second breeding season and argue that this is consistent with age-regulated terminal investment.

A shocking discovery of threat risks on newly described species of weakly electric fishes.

This study aims to investigate relationships between species traits and publication date in the weakly electric osteoglossiform Mormyroidea (African knifefish and elephantfishes) and the ostariophysan Gymnotiformes (Neotropical knifefishes). It is investigated whether body size and geographic distribution area are correlated with publication date and whether extinction risk differs between both phylogenetically distant and geographically isolated clades. Statistical modelling indicates that the number of new species described annually is stable in mormyroids and clearly increasing in gymnotiforms. Best-fitting generalised linear models (GLM) indicate that the newly discovered species are more often of small-bodied, predominantly narrowly distributed and more likely to be threatened with extinction. These characteristics are more pronounced in mormyroids when compared with gymnotiforms, suggesting that some African electric fishes may live an ephemeral existence after formal description. Despite taxonomic work has been more intense in the Neotropics than in Africa in the recent decades, there is evidence that the African continent represents the next frontier of species descriptions. Taxonomic studies are fundamental for the understanding of richness and distribution and hence extinction risk assessment and conservation, of these remarkable convergent fish clades.

Bony Patchwork: Mosaic Patterns of Evolution in the Skull of Electric Fishes (Apteronotidae: Gymnotiformes).

Mosaic evolution refers to the pattern whereby different organismal traits exhibit differential rates of evolution typically due to reduced levels of trait covariation through deep time (i.e., modularity). These differences in rates can be attributed to variation in responses to selective pressures between individual traits. Differential responses to selective pressures also have the potential to facilitate functional specialization, allowing certain traits to track environmental stimuli more closely than others. The teleost skull is a multifunctional structure comprising a complex network of bones and thus an excellent system for which to study mosaic evolution. Here we construct an ultrametric phylogeny for a clade of Neotropical electric fishes (Apteronotidae: Gymnotiformes) and use three-dimensional geometric morphometrics to investigate patterns of mosaic evolution in the skull and jaws. We find strong support for a developmental, three-module hypothesis that consists of the face, braincase, and mandible, and we find that the mandible has evolved four times faster than its neighboring modules. We hypothesize that the functional specialization of the mandible in this group of fishes has allowed it to outpace the face and braincase and evolve in a more decoupled manner. We also hypothesize that this pattern of mosaicism may be widespread across other clades of teleost fishes.

Molecular phylogeny of the ghost knifefishes (Gymnotiformes: Apteronotidae)

Ghost knifefishes (Gymnotiformes: Apteronotidae) are weakly electric fishes that possess a high-frequency, neurogenic electric organ discharge. They are found throughout the humid Neotropics from Panama to Argentina and are most diverse and abundant in the channels of large lowland rivers. Apteronotidae is the most species-rich family of Neotropical electric knifefishes with 96 valid species in 15 genera. We present a phylogenetic hypothesis based on molecular sequence data from three mitochondrial genes (16S, coi, cytb) and four nuclear loci (glyt, rag2, ryr3, zic1). Our analysis includes sequence data for 203 samples in 54 species and 14 genera, making it the most densely-sampled and data-rich phylogeny of the Apteronotidae to date. Our results corroborate previous phylogenetic hypotheses with the placement of Orthosternarchus + Sternarchorhamphus sister to all other apteronotids, a non-monophyletic Apterontotus, and a sister relationship between Sternarchorhynchus and the Navajini. We also report several novel relationships, particularly within the Navajini and among several species of the nominal genus Apteronotus not previously included in phylogenetic analyses. We additionally provide a new classification for the family.

First chromosomal analysis of Gymnorhamphichthys britskii: the remarkable lowest diploid value within the family Rhamphichthyidae (Gymnotiformes)

ABSTRACT Gymnorhamphichthys britskii is a Neotropical electric fish of family Rhamphichthyidae described from the Paraná-Paraguay system. This study reports the first karyotypic description of G. britskii collected from the upper Paraná river basin, which presented 2n=38 chromosomes, karyotype composed of 14 metacentric, 8 submetacentric, 2 subtelocentric and 14 acrocentric chromosomes, and fundamental number as 62 for both sexes. Heteromorphic sex chromosomes were absent. A single pair of nucleolar organizing regions (NORs) was detected in the submetacentric chromosome pair number 9 by silver staining and confirmed by the 18S rDNA probe. The 5S rDNA was located in a single chromosome pair. Heterochromatic regions were clearly observed in the short arms of the NOR-bearing chromosome pair and in the telomeric positions of most acrocentric chromosomes. Besides the present data are valuable to help in understanding karyotypic evolution in Rhamphichthyidae, data from NORs confirmed the tendency of this family in presenting simple NORs sites, similar to the other Gymnotiformes clades. Yet, the presence of a large heterochromatic block in the NOR-bearing chromosome can be used as cytogenetic markers for G. britskii, and that centric fusions appear to be an important mechanism in the karyotype evolution and differentiation among Gymnotiformes species.

Ecomorphology of Neotropical Electric Fishes: An Integrative Approach to Testing the Relationships between Form, Function, and Trophic Ecology

SynopsisThe relationship between form and function is thought to play an integral role in structuring broad-scale patterns of morphological evolution and resource utilization. In ecomorphological studies, mechanical performance is widely understood to constrain the evolution of form and function. However, the relationship between form, function, and resource utilization is less clear. Additionally, seasonal fluctuations in resource availability may further complicate patterns of resource use. How organisms cope with these complexities, and the effect of these factors on broadscale patterns of morphological evolution is also poorly understood. Here we use three-dimensional geometric morphometrics, biomechanics, stable isotope analysis, and gut-content analysis to study trophic evolution in a clade of riverine-adapted electric fishes from a region with high seasonal variability; the Amazon River. We find significant and phylogenetically structured relationships among measures of trophic ecology and skull shape. We also recover a significant relationship between the mechanical advantage of the mandible and trophic position, where species feeding at higher trophic levels have narrower jaws with lower mechanical advantages, and species feeding at lower trophic levels have deeper jaws with higher mechanical advantages. Our results indicate that selection is driving the evolution of mandible shape and performance toward specialization on different trophic ecologies.

Resolving Deep Nodes in an Ancient Radiation of Neotropical Fishes in the Presence of Conflicting Signals from Incomplete Lineage Sorting.

Resolving patterns of ancient and rapid diversifications is one of the most challenging tasks in evolutionary biology. These difficulties arise from confusing phylogenetic signals that are associated with the interplay of incomplete lineage sorting (ILS) and homoplasy. Phylogenomic analyses of hundreds, or even thousands, of loci offer the potential to resolve such contentious relationships. Yet, how much useful phylogenetic information these large data sets contain remains uncertain and often goes untested. Here, we assess the utility of different data filtering approaches to maximize phylogenetic information and minimize noise when reconstructing an ancient radiation of Neotropical electric knifefishes (Order Gymnotiformes) using ultraconserved elements. We found two contrasting hypotheses of gymnotiform evolutionary relationships depending on whether phylogenetic inferences were based on concatenation or coalescent methods. In the first case, all analyses inferred a previously-and commonly-proposed hypothesis, where the family Apteronotidae was found as the sister group to all other gymnotiform families. In contrast, coalescent-based analyses suggested a novel hypothesis where families producing pulse-type (viz., Gymnotidae, Hypopomidae, and Rhamphichthyidae) and wave-type electric signals (viz., Apteronotidae, Sternopygidae) were reciprocally monophyletic. Nodal support for this second hypothesis increased when analyzing loci with the highest phylogenetic information content and further increased when data were pruned using targeted filtering methods that maximized phylogenetic informativeness at the deepest nodes of the Gymnotiformes. Bayesian concordance analyses and topology tests of individual gene genealogies demonstrated that the difficulty of resolving this radiation was likely due to high gene-tree incongruences that resulted from ILS. We show that data filtering reduces gene-tree heterogeneity and increases nodal support and consistency of species trees using coalescent methods; however, we failed to observe the same effect when using concatenation methods. Furthermore, the targeted filtering strategies applied here support the use of "gene data interrogation" rather than "gene genealogy interrogation" approaches in phylogenomic analyses, to extract phylogenetic signal from intractable portions of the Tree of Life.

Why the long face? Static allometry in the sexually dimorphic phenotypes of Neotropical electric fishes

Abstract The evolution of sexually dimorphic traits is thought to have marked effects on underlying patterns of static allometry. These traits can negatively affect organismal survivability by creating trade-offs between trait size and performance. Here we use three-dimensional geometric morphometrics to study the static allometry of two species of sexually dimorphic electric fishes (Apteronotus rostratus and Compsaraia samueli) in which mature males grow elongate jaws used in agonistic male–male interactions. We also estimate jaw-closing performance between the sexes of both species to track changes in kinematic transmission associated with the development of sexual weaponry. We find significantly different patterns of static allometry between the sexes of both species, with males exhibiting more positive allometric slopes relative to females. We also find a negative relationship between skull shape and mandibular kinematic transmission in C. samueli, suggesting a trade-off where males with longer faces exhibit lower mechanical advantages, suggesting weaker jaw leverage. In contrast, males and females of A. rostratus exhibit no difference between sexes in mechanical advantage associated with facial elongation.

Particular Chromosomal Distribution of Microsatellites in Five Species of the Genus Gymnotus (Teleostei, Gymnotiformes).

Microsatellites show great abundance in eukaryotic genomes, although distinct chromosomal distribution patterns might be observed, from small dispersed signals to strong clustered motifs. In Neotropical fishes, the chromosome mapping of distinct microsatellites was employed several times to uncover the origin and evolution of sex and supernumerary chromosomes, whereas a detailed comparative analysis considering different motifs at the chromosomal level is scarce. Here, we report the chromosomal location of several simple sequence repeats (SSRs) in distinct electric knife fishes showing variable diploid chromosome numbers to unveil the structural organization of several microsatellite motifs in distinct Gymnotus species. Our results showed that some SSRs are scattered throughout the genomes, whereas others are particularly clustered displaying intense genomic compartmentalization. Interestingly, the motifs CA, GA, and GAG exhibited a band-like pattern of hybridization, useful for the identification of homologous chromosomes. Finally, the colocalization of SSRs with multigene families is probably related to the association of microsatellites with gene spacers in this case.