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Adaptation to extreme environments often involves the evolution of dramatic physiological changes. To better understand how organisms evolve these complex phenotypic changes, the repeatability and predictability of evolution, and possible constraints on adapting to an extreme environment, it is important to understand how adaptive variation has evolved. Poeciliid fishes represent a particularly fruitful study system for investigations of adaptation to extreme environments due to their repeated colonization of toxic hydrogen sulfide-rich springs across multiple species within the clade. Previous investigations have highlighted changes in the physiology and gene expression in specific species that are thought to facilitate adaptation to hydrogen sulfide-rich springs. However, the presence of adaptive nucleotide variation in coding and regulatory regions and the degree to which convergent evolution has shaped the genomic regions underpinning sulfide tolerance across taxa are unknown. By sampling across seven independent lineages in which nonsulfidic lineages have colonized and adapted to sulfide springs, we reveal signatures of shared evolutionary rate shifts across the genome. We found evidence of genes, promoters, and putative enhancer regions associated with both increased and decreased convergent evolutionary rate shifts in hydrogen sulfide-adapted lineages. Our analysis highlights convergent evolutionary rate shifts in sulfidic lineages associated with the modulation of endogenous hydrogen sulfide production and hydrogen sulfide detoxification. We also found that regions with shifted evolutionary rates in sulfide spring fishes more often exhibited convergent shifts in either the coding region or the regulatory sequence of a given gene, rather than both.

Predation threat is a major driver of behavior in many prey species. Animals can recognize their relative risk of predation based on cues in the environment, including visual and/or chemical cues released by a predator or from its prey. When threat of predation is high, prey often respond by altering their behavior to reduce their probability of detection and/or capture. Here, we test how a clonal fish, the Amazon molly (Poecilia formosa), behaviorally responds to predation cues. We measured aggressive and social behaviors both under 'risk', where chemical cues from predatory fish and injured conspecifics were present, and control contexts (no risk cues present). We predicted that mollies would exhibit reduced aggression towards a simulated intruder and increased sociability under risk contexts as aggression might increase their visibility to a predator and shoaling should decrease their chance of capture through the dilution effect. As predicted, we found that Amazon mollies spent more time with a conspecific when risk cues were present, however they did not reduce their aggression. This highlights the general result of the 'safety in numbers' behavioral response that many small shoaling species exhibit, including these clonal fish, which suggests that mollies may view this response as a more effective anti-predator response compared to limiting their detectability by reducing aggressive conspecific interactions.

This study examines the impact of two types of environmental enrichment on the growth of the Amazon molly (Poecilia formosa), a clonal fish species. We investigated the effects of two welfare-related enrichment factors: physical enrichment (presence of a halfpipe PVC and gravel substrate, E) and social enrichment (presence of visible neighbor fish, N). Fish were divided into four treatment groups: (1) both physical and social enrichment (EN), (2) no physical enrichment but social enrichment (nEN), (3) physical enrichment without social enrichment (EnN,), and (4) no enrichment (nEnN). Results showed that individuals, which received both types of enrichment, exhibited faster growth rates and lower early life mortality compared to the other groups, indicating improved health and welfare. However, this initial advantage did not persist into adulthood, suggesting that the benefits of enrichment may be stage specific. These findings highlight the importance of environmental factors in promoting welfare in a particular species, especially during early developmental stages. This study may have implications for early life-stage fish research indicating that some environmental factors may play a role in enhancing growth, reproductive competence, and reduced mortality. Enrichment effects on growth, health, and reproduction in a single clone of the asexual Amazon molly, Poecilia formosa. [ABSTRACT FROM AUTHOR]

"El Aerolito" is the most biodiverse cenote cave globally, home to Poecilia velifera and Gambusia yucatana. However, their trophic niches and interactions remain undescribed. A total of 53 P. velifera and 38 G. yucatana were analyzed for stable isotopes, along with water samples for chlorophyll, silicates, and physical–chemical parameters. Data were collected during the norths, rainy, and dry seasons in the year 2023. Isotopic niches were calculated, and a PCA analysis was used to assess environmental factors. P. velifera showed δ13C values of −25.4‰, −25.7‰, and −24.5‰, whereas G. yucatana exhibited −24.8‰, −24.0‰, and −24.2‰ across seasons. Niche size varied, with the largest during the norths and smallest during the rainy season, indicating seasonality. PCA linked environmental parameters (e.g., temperature and dissolved oxygen) to resource availability, suggesting niche shifts due to rainfall. Mangroves and insects contributed to the isotopic signals, with shifts observed during the dry and rainy seasons. The highest water parameter values occurred during the dry and rainy seasons, while the lowest was during the norths. The system, nutrient-poor and lacking chlorophyll production, relies on external sources. The study concludes that isotopic niches are dynamic and seasonally influenced by abiotic factors, especially rainfall. [ABSTRACT FROM AUTHOR]

Context: Spinal cord injury (SCI) is a devastating condition that results in severe disability and significant comorbidities. The complex pathophysiology of SCI repair and difficulties understanding neural regeneration are treatment challenges. Objectives: The aim of this study is to systematically review the diverse applications of various fish species as models for SCI research. Evidence Acquisition: PRISMA guidelines were used to review observational and interventional studies that utilized fish species as SCI models, published from inception to July 2023. Two independent reviewers screened and performed the data extraction. One independent reviewer assessed the risk of bias for the included studies. Results: Five thousand six hundred and thirty-three records were reviewed, and 144 met the inclusion criteria and were categorized by fish species. The majority of studies employed complete transection injuries, with the remainder being crush injuries, laser injuries, electro-ablations, and demyelination with substances. Zebrafish (Danio rerio) were most commonly utilized 102/144 (71%), primarily with larval models. Other models included Lamprey (Petromyzon marinus and Lethenteron reissued); Goldfish (Carassius auratus); European eel (Anguilla Anguilla); Knifefish (Apteronotus leptorhynchus and Apteronotus albifrons); Sailfin Molly (Poecilia latipinna); and African turquoise killifish (Nothobranchiusfurzeri). Conclusions: This systematic review highlights that fish models, particularly zebrafish, goldfish, and European eels, are important models for further defining SCI pathophysiology and regenerative processes. These models provide a less complex model to gain insights into apoptosis and glial networks. [ABSTRACT FROM AUTHOR]

Background: It is widely reported that exotic larvicidal fishes like Gambusia and Poecilia have adverse impact on the native aquatic fauna. The present study highlights the efficacy of two colourful native murrels, primarily designated as ornamental fish, Channa bleheri and Channa stewartii as biocontrol agent of mosquito larvae. Methods: Live specimens of Channa bleheri and C. stewartii, collected from the wetlands of Tinsukia district of Upper Assam, were assessed for their larvivorous potential at individual and group levels during day and night by dividing the specimens into two size groups. After 12 hour and 24 hour starvation, the test specimens (mean size for small group 8.67-9.17 cm and that of large group 11.63-13.27 cm) were given known number of mosquito larvae and recorded the consumption rate. Result: The predation rate varied from 33.3±4.36 to 71.6±5.15/min for Channa bleheri and that of C. stewartii from 16.3±0.95 to 68.2±2.77/min. In both species, smaller sized specimens were better performers as predators. Predation rate at 12 and 24 hrs of starvation and between day and night proved that these native murrels are excellent predators of mosquito larvae. [ABSTRACT FROM AUTHOR]

Coloration facilitates evolutionary investigations in nature because the interaction between genotype, phenotype and environment is relatively accessible. In a landmark set of studies, Endler addressed this complexity by demonstrating that the evolution of male Trinidadian guppy coloration is shaped by the local balance between selection for mate attractiveness versus crypsis. This became a textbook paradigm for how antagonistic selective pressures may determine evolutionary trajectories in nature. However, recent studies have challenged the generality of this paradigm. Here, we respond to these challenges by reviewing five important yet underappreciated factors that contribute to colour pattern evolution: (i) among-population variation in female preference and correlated variation in male coloration, (ii) differences in how predators versus conspecifics view males, (iii) biased assessment of pigmentary versus structural coloration, (iv) the importance of accounting for multi-species predator communities, and (v) the importance of considering the multivariate genetic architecture and multivariate context of selection and how sexual selection encourages polymorphic divergence. We elaborate these issues using two challenging papers. Our purpose is not to criticize but to point out the potential pitfalls in colour research and to emphasize the depth of consideration necessary for testing evolutionary hypotheses using complex multi-trait phenotypes such as guppy colour patterns.

Evaluating the fitness of hybrids can provide important insights into genetic differences between species or diverging populations. We focused on surface- and cave-ecotypes of the widespread Atlantic molly Poecilia mexicana and raised F1 hybrids of reciprocal crosses to sexual maturity in a common-garden experiment. Hybrids were reared in a fully factorial 2 × 2 design consisting of lighting (light vs. darkness) and resource availability (high vs. low food). We quantified survival, ability to realize their full reproductive potential (i.e. completed maturation for males and 3 consecutive births for females) and essential life-history traits. Compared to the performance of pure cave and surface fish from a previous experiment, F1s had the highest death rate and the lowest proportion of fish that reached their full reproductive potential. We also uncovered an intriguing pattern of sex-specific phenotype expression, because male hybrids expressed cave molly life histories, while female hybrids expressed surface molly life histories. Our results provide evidence for strong selection against hybrids in the cave molly system, but also suggest a complex pattern of sex-specific (opposing) dominance, with certain surface molly genes being dominant in female hybrids and certain cave molly genes being dominant in male hybrids. [ABSTRACT FROM AUTHOR]

The role of hybridization as a formative process in evolution has received much attention in the past few decades. A particularly fascinating outcome of hybrid speciation is the formation of asexual hybrid species. The Amazon molly (Poecilia formosa) is such a hybrid and originated from a P. mexicana mother and a P. latipinna father. Consequently, a heterospecific mating must have occurred leading to the Amazon molly, indicating a breakdown of any potential prezygotic isolation between parental species. Here we studied the female mate preferences of extant P. mexicana and P. latipinna from several populations using standard binary choice tests with males of both sexual species that were matched for size. Poecilia mexicana and P. latipinna can be crossed in the lab, however, the offspring are not asexual, but sexual F1s. In our study, we generated F1s and tested their mating preferences with sexual males of both P. mexicana and P. latipinna against F1 males. Overall, our results show that in extant P. mexicana and P. latipinna no female preference for conspecific males was detectable. Consequently, heterospecific matings are possible and not hindered by any apparent behavioral prezygotic isolation. If female preferences in these species were comparable around the time the Amazon molly originated as a hybrid species ca. 100,000 years ago, matings leading to hybrids would be very likely. F1 females also have no discernable mating preferences for either sexual males or F1 males. Such lack of prezygotic behavioral isolation could potentially lead to F2 individuals, backcrosses, and introgression. [ABSTRACT FROM AUTHOR]

Phenotypic plasticity, the ability of a single genotype to produce different phenotypes under different environmental conditions, plays a profound role in several areas of evolutionary biology. One important role is as an adaptation to a variable environment. While plasticity is extremely well documented in response to many environmental factors, there is controversy over how much of that plasticity is adaptive. Evidence is also mixed over how often conspecific populations display qualitative differences in the nature of plasticity. We present data on the reaction norms of growth and maturation to variation in temperature and salinity in male and female sailfin mollies (Poecilia latipinna) from three locally adjacent populations from South Carolina (SC). We compare these reaction norms to those previously reported in locally adjacent populations from north Florida (NF). In general, patterns of plasticity in fish from SC were similar to those in fish from NF. The magnitude of plasticity differed; fish from SC displayed less plasticity than fish from NF. This was because SC fish grew faster and matured earlier at the lower temperatures and salinities compared to NF fish. This is a countergradient pattern of variation, in which SC fish grew faster and matured earlier in conditions that would otherwise slow growth and delay maturity. Among fish from both regions, males were much less plastic than females, especially for length at maturity. While there was no detectable heterogeneity among populations from NF, males from one of the SC populations, which is furthest from the other two, displayed a qualitatively different response in age at maturity to temperature variation than did males from the other two SC populations. The pattern of population variation in plasticity within and among regions suggests that gene flow, which diminishes with distance in sailfin mollies, plays a critical role in constraining divergence in norms of reaction. [ABSTRACT FROM AUTHOR]

Fish live in aquatic areas with varying salinity from fresh water to seawater. Gills are one of the most important osmoregulatory organs for maintaining the salt–water balance. In this study, changes in the gills of velifera (Poecilia sphenops) acclimated to 20, 30, 40, and 50 ppt of saline in the laboratory were examined histologically and immunohistochemically. Mucus cells (MCs), proliferating cell nuclear antigen (PCNA), mitochondria-rich cells (MRCs), and heat shock proteins (HSP70) + cells were marked in the gill tissue, which underwent routine histological procedures, and the severity of histological alterations increase with the increase of the salinity. When the gills were examined histologically, it was determined that there was an increase in cells due to hyperplasia, especially in the primary lamellae. A rapid increase in MC and HSP70 + cell numbers was observed in the 20 and 30 ppt groups. Although PCNA + cells were present in each salinity group, they were observed in the salinity group up to 50 ppt. An increase in the number and volume of MRCs was observed with increasing salinity. Based on the histological and immunohistochemical results, it is thought that the cytological and molecular arrangements of cells in gills of sailfin velifera under different levels of salinity are essential for the regulation of ion and salt balance. [ABSTRACT FROM AUTHOR]

Mucuna pruriens, a rich source of L-dihydroxyphenylalanine, commonly known as L-DOPA and a precursor to dopamine, holds potential as a natural nutritional supplement. This study aimed to delve into the impact of incorporating M. pruriens seed powder (MpSP) into the feed on growth parameters and gonadal development of mono-sex common molly (Poecilia sphenops). The fish population was divided into three experimental groups, such as G1, G2, and G3, and a control group (C), each comprising 20 individuals. Over 45 days, the experimental groups were nourished with a commercial diet bolstered by MpSP in different concentrations (5, 7 and 10g/kg of feed, respectively). In contrast, the control group was provided with a regular diet devoid of the supplement. At the end of the experiment, MpSP demonstrated significant modulation (p<0.05) of growth performance metrics, including specific growth rate (SGR), length gain rate (LGR), body mass gain (BMG), and feed conversion ratio (FCR). Impressively, even the lower concentration of MpSP (5g/kg diet) yielded substantial increments in sperm count (p <0.05) and gonadosomatic index (GSI). These findings were corroborated by histological changes that reflected enhanced testicular development, consistently outperforming the control group. These outcomes collectively suggest the potential of velvet bean seed powder as a feasible, natural, and costeffective dietary supplement for enhancing growth and testicular development in mono-sex P. sphenops. [ABSTRACT FROM AUTHOR]

Teratoma is a rare tumour in fish consisting of tissues from more than one germ layer, that may be located in either the gonads or extragonadal sites. Teratomas in many fish species remain poorly understood. In this work, we performed the first histological examinations of extragonadal teratomas in Poecilia wingei and also examined the influence of a large teratoma on male sexual activity. The studied teratomas shared general organizational features, but they also had variations in both external and internal features. In teratomas, the most common and highly differentiated tissues were striated muscle and nervous tissue. Despite the tumour, the male P. wingei exhibited normal mating behaviour and retained the ability for successful copulation. The structural features of extragonadal teratomas in guppy fish indicate a possible connection between these tumours and a failure of conserved processes operating in the embryonic germline. [ABSTRACT FROM AUTHOR]

In order to search of viable alternative protein source for aquaculture, a feeding trial of six-months was carried out to test the effect of mealworm (Tenebrio molitor) meal relacing fishmeal protein at 0% (control), 25%, 50%, 75% and 100% in practical diets (named D0, D25, D50, D75 and D100 respectively) on the growth performance, feed utilization and survival of black molly (Poecilia sphenops). At the end of six-months, significantly higher final weight and length were observed in D75 diet fed set followed by D50 set and then in D0 set, whereas lowest value was recorded in D100. The specific growth rate (SGR) in percent of control fishes (D0) was 2.25±0.02, whereas the result of SGR (%) was noted best in D75 set (2.44±0.02) and poorest in the D100 (1.97±0.02). Although D75 diet fed set showed lower feed conversion ratio (FCR), black molly fed with D25 and D50 diets still utilized the diets as well as control (D0) as their values did not vary significantly. Significantly higher survival percentage was 96.67%, observed in D75 set, in contrast lowest found in D100 set (64.44%). Hence, mealworm could be successfully incorporated by replacing fishmeal from 50% to 75% in the diet of black molly with optimal growth, feed utilization and better survival. [ABSTRACT FROM AUTHOR]

Visual sensitivity and body pigmentation are often shaped by both natural selection from the environment and sexual selection from mate choice. One way of quantifying the impact of the environment is by measuring how traits have changed after colonization of a novel habitat. To do this, we studied Poecilia mexicana populations that have repeatedly adapted to extreme sulphidic (H2 S-containing) environments. We measured visual sensitivity using opsin gene expression, as well as body pigmentation, for populations in four independent drainages. Both visual sensitivity and body pigmentation showed significant parallel shifts towards greater medium-wavelength sensitivity and reflectance in sulphidic populations. Altogether we found that sulphidic habitats select for differences in visual sensitivity and pigmentation. Shifts between habitats may be due to both differences in the water's spectral properties and correlated ecological changes.

Behavioral individuality is a ubiquitous phenomenon in animal populations, yet the origins and developmental trajectories of individuality, especially very early in life, are still a black box. Using a high-resolution tracking system, we mapped the behavioral trajectories of genetically identical fish (Poecilia formosa), separated immediately after birth into identical environments, over the first 10 weeks of their life at 3 s resolution. We find that (i) strong behavioral individuality is present at the very first day after birth, (ii) behavioral differences at day 1 of life predict behavior up to at least 10 weeks later, and (iii) patterns of individuality strengthen gradually over developmental time. Our results establish a null model for how behavioral individuality can develop in the absence of genetic and environmental variation and provide experimental evidence that later-in-life individuality can be strongly shaped by factors pre-dating birth like maternal provisioning, epigenetics and pre-birth developmental stochasticity.

Three experiments were conducted examining whether an artificially selected "gold" color variant in female "models" affects mate choice copying behavior in sailfin mollies (Poecilia latipinna). Experiment I consisted of a pair of female preference assays, first assessing preference for male body size, followed by a mate choice copying assay that paired a model female with the smaller, non-preferred male from the initial preference test. Female subjects were divided into three groups that used either a wildtype female model, an artificially selected "gold" variant (cultivated within the aquarium fish trade) model, or control wherein no model was presented. Results showed females consistently copied the model's choice, switching preferences from the larger to smaller male when paired with a model regardless of color. In the second experiment wildtype females were presented with a pair of size-matched dummy males both of which paired with model females (one gold and the other wild type). Subjects consistently preferred the male previously paired with the gold- over the male with the wildtype-model, suggesting pre-existing sensory/perceptual biases may have affected their mate choice copying behavior. Previous studies have offered evidence for the spread of novel traits in males via sensory exploitation. However, these results indicate such biases may influence courtship behavior in circumstances where the novel trait is expressed in females as well. For the third experiment, wildtype females were presented with a choice between gold vs wildtype dummy males, the results of which revealing significant preferences for gold. In a follow-up assay pairing a wild type model with the non-preferred wildtype male, females maintained their preference for gold males despite the conflicting social driver of mate choice copying. These data offer evidence for the existence of a perceptual/cognitive bias in the context of mate choice copying, favoring the gold phenotype and/or novelty in general. [ABSTRACT FROM AUTHOR]

Simple Summary: This study scrutinized the retinal composition of Molly fish (Poecilia sphenops), revealing a complex neuronal structure. The retina showcased a distinctive square mosaic layout of cones, incorporating double cones alongside two variants of single cones. Within the inner nuclear layer, a diverse array of cells and Müller cell processes, expressing GFAP, traversed the retina. Notably, astrocyte cell processes expressing GFAP were discernible in both the inner and outer plexiform layers. Overall, Molly fish's retina displayed a sophisticated structure characterized by a high density of photoreceptors, bipolar, amacrine, horizontal, Müller cells, and astrocytes, suggesting robust photopic visual capabilities. The specific arrangement and distribution of photoreceptors in the retina can vary among different fish species, with each species exhibiting adaptations related to its habitat, behavior, and visual requirements. Poecilia sphenops, a diurnal fish, was the focus of this study. The retinas of a total of eighteen Molly fish were investigated utilizing light and electron microscopy. The retina exhibited a square mosaic pattern of the inner segments of cones. This pattern comprised double cones positioned along the sides of a square, with two types of single cones situated at the center and corners of the square arrangement across the entire retina. The corner cones were slightly shorter than the central ones. Additionally, the outer plexiform layer contained both cone pedicles and rod spherules. The rod spherule consisted of a single synaptic ribbon arranged in a triad or quadrat junctional arrangement within the invaginating free ends of the horizontal and bipolar cell processes. On the other hand, cone pedicles have more than one synaptic ribbon in their junctional complex. The inner nuclear layer consisted of the amacrine, bipolar, Müller, and horizontal cell bodies. Müller cell processes, expressing GFAP, extended across all retinal layers, segmenting the deeper retina into alternating fascicles of optic axons and ganglion cells. The outer and inner plexiform layers showed many astrocyte cell processes expressing GFAP. In conclusion, the current study is the first record of the retinal structures of Molly fish. This study illustrated the mosaic arrangement of photoreceptors and GFAP expression patterns of astrocytes and Müller cells. The presence of three cone types, coupled with a sufficient number of rods, likely facilitates motion awareness for tasks like finding food and performing elaborate mating ceremonies. [ABSTRACT FROM AUTHOR]

Behavioural plasticity allows organisms to respond to environmental challenges on short time scales. But what are the ecological and evolutionary processes that underlie behavioural plasticity? The answer to this question is complex and requires experimental dissection of the physiological, neural and molecular mechanisms contributing to behavioural plasticity as well as an understanding of the ecological and evolutionary contexts under which behavioural plasticity is adaptive. Here, we discuss key insights that research with Trinidadian guppies has provided on the underpinnings of adaptive behavioural plasticity. First, we present evidence that guppies exhibit contextual, developmental and transgenerational behavioural plasticity. Next, we review work on behavioural plasticity in guppies spanning three ecological contexts (predation, parasitism and turbidity) and three underlying mechanisms (endocrinological, neurobiological and genetic). Finally, we provide three outstanding questions that could leverage guppies further as a study system and give suggestions for how this research could be done. Research on behavioural plasticity in guppies has provided, and will continue to provide, a valuable opportunity to improve understanding of the ecological and evolutionary causes and consequences of behavioural plasticity. [ABSTRACT FROM AUTHOR]

This research study explores the effects of Tribulus terrestris on the testicular enzyme activities in Poecilia sphenops, (a fish species) with the aim of reproductive manipulation. The study entails incorporating the plant extract into preprepared fish feed, which is then administered to the fish. A comprehensive phytochemical and Gas Chromatography-Mass Spectrometry (GC-MS) analysis was conducted to quantitatively analyze the phytochemical composition in the Tribulus Terrestris extract. The fish feed was prepared using various concentrations of Tribulus terrestris and provided for Poecilia sphenops. The fish were observed and analyzed for three months. The fish's growth performance and body composition were assessed. Subsequently, the testis and liver were individually homogenized and subjected to centrifugation to obtain enzyme extracts for analysis. The testicular enzyme activities of lactate dehydrogenase (LDH), sorbitol dehydrogenase (SDH), acid phosphatases (ACP) and alkaline phosphatase (ALP) were evaluated as markers for variations in testicular function, cellular stress and energy metabolism. The study determined that the incorporation of different concentrations of Tribulus terrestris into the feed improved the rate of growth, overall health and reproductive success of Poecilia sphenops. The findings provide helpful insights into the potential influence of Tribulus terrestris on the reproductive behaviors of fish. [ABSTRACT FROM AUTHOR]

Environmental factors can promote phenotypic variation through alterations in the epigenome and facilitate adaptation of an organism to the environment. Although hydrogen sulfide is toxic to most organisms, the fish Poecilia mexicana has adapted to survive in environments with high levels that exceed toxicity thresholds by orders of magnitude. Epigenetic changes in response to this environmental stressor were examined by assessing DNA methylation alterations in red blood cells, which are nucleated in fish. Males and females were sampled from sulfidic and nonsulfidic natural environments; individuals were also propagated for two generations in a nonsulfidic laboratory environment. We compared epimutations between the sexes as well as field and laboratory populations. For both the wild-caught (F0) and the laboratory-reared (F2) fish, comparing the sulfidic and nonsulfidic populations revealed evidence for significant differential DNA methylation regions (DMRs). More importantly, there was over 80% overlap in DMRs across generations, suggesting that the DMRs have stable generational inheritance in the absence of the sulfidic environment. This is an example of epigenetic generational stability after the removal of an environmental stressor. The DMR-associated genes were related to sulfur toxicity and metabolic processes. These findings suggest that adaptation of P. mexicana to sulfidic environments in southern Mexico may, in part, be promoted through epigenetic DNA methylation alterations that become stable and are inherited by subsequent generations independent of the environment.

Throughout their distribution range, sailfin and shortfin mollies of the genus Poecilia inhabit overlapping habitats; however, little is known about its behavioural coexistence. We first describe the ethogram of Poecilia velifera and P. mexicana in a mangrove of Yucatan. Furthermore, we identify behavioural strategies of each size morphotype/species. We describe reproductive, territorial, and cleaning behaviours in two sympatric species of mollies in their natural context, with emphasis of their behavioural interactions and coexistence. Specifically, here we document (i) specific reproductive strategies in male P. velifera morphotypes (large, intermediate-ornamented and 'cryptic-intermediate'), suggesting that cryptic-intermediate males have advantages that suggest consequences for their reproductive success; (ii) frequent courtship and sexual behaviours from P. mexicana males directed to P. velifera females, that deserve future attention to determine the possible generation of hybrids; and (iii) increased aggressive behaviour and territoriality of P. mexicana relative to those observed in P. velifera. [ABSTRACT FROM AUTHOR]

Plastic pollution is now a ubiquitous feature of freshwater systems and the majority of this is fibrous. Here, we test the effects of plastic and cellulose-based fibres (polyester, cotton, and bamboo from commercial clothing) on fish host-parasite interactions using a freshwater fish host-parasite model system (guppy Poecilia reticulata-Gyrodactylus turnbulli). For uninfected fish, polyester exposure was associated with significantly higher mortality rates compared with the other two fibre types. For infected fish, whilst polyester and cotton exposure were not associated with any significant changes to parasite burdens, fish exposed to bamboo fibres had significantly reduced maximum parasite burdens compared with fish not exposed to any fibres, indicating that the bamboo fibres and/or associated dyes conferred some degree of resistance or tolerance. Whilst unable to determine the exact nature of the chemical dyes, when testing off-host parasite survival on exposure to the fibre dyes, cotton and particularly polyester dyes were associated with higher parasite mortality compared to bamboo. Overall, we add to the growing body of evidence which shows that polyester microplastic fibres and their associated dyes can be detrimental for both fish and parasite survival, and we highlight the need for increased transparency from textile industries on the chemical identity of fabric dyes. [ABSTRACT FROM AUTHOR]

The Amazon Molly (Poecilia formosa) reproduces by gynogenesis, a relatively rare form of asexual reproduction where sperm is required to trigger embryogenesis, but male genes are not incorporated into the genome of the embryo. Studying gynogenesis could isolate paternal non-genetic effects on reproduction. This study explored which of eleven related species can produce sperm to trigger gynogenesis through natural mating in P. formosa, and whether sympatry affects reproductive success in P. formosa. Reproductive outcomes measured were relative reproductive output (number of offspring in the first brood divided by female standard length), relative embryo output (number of embryos in the first brood divided by female standard length) and combined relative reproductive output (sum of relative reproductive output and relative embryo output). For large (>4 cm) P. formosa, combined relative reproductive output was higher with sympatric Atlantic Molly (Poecilia mexicana) males than with allopatric P. mexicana males. P. formosa produced live offspring or late-stage embryos with all species tested in the genera Poecilia and Limia but did not produce offspring or embryos with males from the genera Gambusia, Girardinus, Heterandria, Poeciliopsis, or Xiphophorus. This information, as well as the limitations characterized in this study, will set a foundation for use of P. formosa as a model for paternal effects and the species specificity of sperm on fertilization, embryogenesis, and reproductive success. [ABSTRACT FROM AUTHOR]

Extreme environments test the limits of life; yet, some organisms thrive in harsh conditions. Extremophile lineages inspire questions about how organisms can tolerate physiochemical stressors and whether the repeated colonization of extreme environments is facilitated by predictable and repeatable evolutionary innovations. We identified the mechanistic basis underlying convergent evolution of tolerance to hydrogen sulfide (H2S)-a toxicant that impairs mitochondrial function-across evolutionarily independent lineages of a fish (Poecilia mexicana, Poeciliidae) from H2S-rich springs. Using comparative biochemical and physiological analyses, we found that mitochondrial function is maintained in the presence of H2S in sulfide spring P. mexicana but not ancestral lineages from nonsulfidic habitats due to convergent adaptations in the primary toxicity target and a major detoxification enzyme. Genome-wide local ancestry analyses indicated that convergent evolution of increased H2S tolerance in different populations is likely caused by a combination of selection on standing genetic variation and de novo mutations. On a macroevolutionary scale, H2S tolerance in 10 independent lineages of sulfide spring fishes across multiple genera of Poeciliidae is correlated with the convergent modification and expression changes in genes associated with H2S toxicity and detoxification. Our results demonstrate that the modification of highly conserved physiological pathways associated with essential mitochondrial processes mediates tolerance to physiochemical stress. In addition, the same pathways, genes, and-in some instances-codons are implicated in H2S adaptation in lineages that span 40 million years of evolution.

Eye regression occurs across cave-dwelling populations of many species and is often coupled with a decrease or loss in eye function. Teleost fishes are among the few vertebrates to undergo widespread colonization of caves and often exhibit eye regression with blindness. Cave populations of the poeciliid fish Poecilia mexicana (cave molly) exhibit reduced-albeit functional-eyes, offering the opportunity to investigate partial eye regression. We sequenced eye transcriptomes of cave and surface populations of P. mexicana to identify differentially expressed genes that potentially underlie eye regression in cave mollies. We identified 28 significantly differentially expressed genes, 20 of which were directly related to light sensitivity, eye structure and visual signaling. Twenty-six of these genes were downregulated in cave compared to surface populations. Functional enrichment analysis revealed eye-related gene ontologies that were under-represented in cave mollies. In addition, a set of co-expressed genes related to vision and circadian rhythm was correlated with habitat type (cave versus surface). Our study suggests that differential gene expression plays a key role in the beginning evolutionary stages of eye regression in P. mexicana, shedding further light on regressive evolution in cavefish.

The molecular basis of convergent phenotypes is often unknown. However, convergence at a genomic level is predicted when there are large population sizes, gene flow among diverging lineages or strong genetic constraints. We used whole-genome resequencing to investigate genomic convergence in fishes ( Poecilia spp.) that have repeatedly colonized hydrogen sulfide (H2S)-rich environments in Mexico. We identified genomic similarities in both single nucleotide polymorphisms (SNPs) and structural variants (SVs) among independently derived sulfide spring populations, with approximately 1.2% of the genome being shared among sulfidic ecotypes. We compared these convergent genomic regions to candidate genes for H2S adaptation identified from transcriptomic analyses and found that a significant proportion of these candidate genes (8%) were also in regions where sulfidic individuals had similar SNPs, while only 1.7% were in regions where sulfidic individuals had similar SVs. Those candidate genes included genes involved in sulfide detoxification, the electron transport chain (the main toxicity target of H2S) and other processes putatively important for adaptation to sulfidic environments. Regional genomic similarity across independent populations exposed to the same source of selection is consistent with selection on standing variation or introgression of adaptive alleles across divergent lineages. However, combined with previous analyses, our data also support that adaptive changes in mitochondrially encoded subunits arose independently via selection on de novo mutations. Pressing questions remain on what conditions ultimately facilitate the independent rise of adaptive alleles at the same loci in separate populations, and thus, the degree to which evolution is repeatable or predictable. This article is part of the theme issue 'Convergent evolution in the genomics era: new insights and directions'.

Accurate knowledge of species colour discrimination is fundamental to explain colour based behaviours and the evolution of colour patterns. We tested how the receptor noise limited model, widely used in behavioural ecology, matched actual colour discrimination thresholds obtained using behavioural tests. Guppies (Poecilia reticulata) were first trained to push a target coloured disk placed among eight grey disks of various luminances on a grey plate. Guppies were then tested to find target disks, which varied in colour contrast from the plate. The target disks followed a gradient going from high contrast to inconspicuous against the grey background. We plotted the percentage of correct choices of each colour in the gradient against the model prediction and determined the discrimination thresholds using the inflection point of the fitted sigmoid curve. We performed the experiment on six colour gradients: red, orange, yellow, green, blue and purple. Four colour gradients: red, orange, green and blue, showed a discrimination threshold that matched the model predictions. However, deviations of the model for the yellow and purple gradients suggest that ecological relevance of some colours could affect decision-making in behavioural tests and that we can no longer assume that the rules for colour discrimination are independent of colours.

Visual pigments can vary across the retina in many vertebrates, but the behavioural consequences of this retinal heterogeneity are unknown. Guppies (Poecilia reticulata) vary dorsoventrally in visual pigments and forage both on the ground and at the water surface, exposing different retinal regions to two very different visual environments. We tested guppy behaviour towards a moving stimulus presented below or above the guppy. We used 12 different narrow-band wavelength stimuli matching each of the opsin peak sensitivities presented either at the top or the bottom of our experimental apparatus. We analysed behaviours of 50 male and 50 female guppies over 4800 trials where a moving stimulus pattern was presented to each guppy. We found that wavelength, position and speed of the stimuli influenced male and female behaviour and seems to be mediated by the long wavelength sensitive photoreceptors. Males also had stronger behavioural responses than females whereas females performed more foraging-related pecking behaviour. Our results suggest that the spatial requirement of visual tasks and their ecological context are important and appear to be partly correlated with photoreceptor arrangement in the retina.

Twenty-four adult molly fish (Poecilia sphenops, Valenciennes 1846) were collected to study the morphology and distribution of ganglia using histological, immunohistochemical, and electron microscopy and focusing on their relation to the immune cells. The ganglia were classified spatially into cranial and spinal, and functionally into sensory and autonomic. Spinal ganglia (dorsal root ganglia, DRG) contained large close ganglionic cells, enclosed by satellite cells, as well as bundles of both myelinated and non-myelinated nerve fibers. There are glial cells, immune cells and telocytes close to the ganglion. In addition, oligodendrocytes were closely related to myelinated axons. Glial fibrillary acidic protein (GFAP) expression was confined to the glia cells and the nerve fibers in the cervical ganglia next to the gills, and surprisingly, in the large ganglionic cells of the DRG. The vestibular ganglia were large, connected to the hind brain, and contained numerous neurons packed in columns. The cervical ganglia were large and observed around the pseudobranch, head kidney, and thymus. Their neurons are randomly distributed, and nerve fibers are peripherally situated. CD3-positive T-lymphocytes, dendritic cells, and CD68-positive macrophages were in close contact with the ganglia. Furthermore, the ganglia around the head kidney showed positive Iba1-expressing cells. Most ganglion cells and nerve fibers in the DRG, autonomic, and vestibular ganglia showed moderate to strong S-100 immunoreactivity. The enteric glia, CD68-expressing macrophages, and acetylcholine (Ach)-expressing neurons were observed along the muscular layer of the intestinal wall. In conclusion, different ganglia of molly fish displayed direct communication with immune cells which support and maintain healthy ganglionic cells. [ABSTRACT FROM AUTHOR]

The pseudobranch is a gill-like structure that exhibits great variations in structure and function among fish species, and therefore, it has remained a topic of investigation for a long time. This study was conducted on adult Molly fish (Poecilia sphenops) to investigate the potential functions of their pseudobranch using histological, histochemical, immunohistochemical analysis, and scanning electron microscopy. The pseudobranch of Molly fish was of embedded type. It comprised many rows of parallel lamellae that were fused completely throughout their length by a thin connective tissue. These lamellae consisted of a central blood capillary, surrounded by large secretory pseudobranch cells (PSCs). Immunohistochemical analysis revealed the expression of PSCs for CD3, CD45, iNOS-2, and NF-κB, confirming their role in immunity. Furthermore, T-lymphocytes-positive CD3, leucocytes-positive CD45, and dendritic cells-positive CD-8 and macrophage- positive APG-5 could be distinguished. Moreover, myogenin and TGF-β-positive PSCs were identified, in addition to nests of stem cells- positive SOX-9 were detected. Melanocytes, telocytes, and GFAP-positive astrocytes were also demonstrated. Scanning electron microscopy revealed that the PSCs were covered by microridges, which may increase the surface area for ionic exchange. In conclusion, pseudobranch is a highly specialized structure that may be involved in immune response, ion transport, acid–base balance, as well as cell proliferation and regeneration. [ABSTRACT FROM AUTHOR]

Collective behaviour is widely accepted to provide a variety of antipredator benefits. Acting collectively requires not only strong coordination among group members, but also the integration of among-individual phenotypic variation. Therefore, groups composed of more than one species offer a unique opportunity to look into the evolution of both mechanistic and functional aspects of collective behaviour. Here, we present data on mixedspecies fish shoals that perform collective dives. These repeated dives produce water waves capable of delaying and/or reducing the success of piscivorous bird attacks. The large majority of the fish in these shoals consist of the sulphur molly, Poecilia sulphuraria, but we regularly also found a second species, the widemouth gambusia, Gambusia eurystoma, making these shoals mixed-species aggregations. In a set of laboratory experiments, we found that gambusia were much less inclined to dive after an attack as compared with mollies, which almost always dive, though mollies dived less deep when paired with gambusia that did not dive. By contrast, the behaviour of gambusiawas not influenced by the presence of diving mollies. The dampening effect of less responsive gambusia on molly diving behaviour can have strong evolutionary consequences on the overall collective waving behaviour as we expect shoals with a high proportion of unresponsive gambusia to be less effective at producing repeated waves. [ABSTRACT FROM AUTHOR]

Mexico is a megadiverse region with a complex geological history, but it remains unclear to what extent the distribution of freshwater fish has been influenced by geographic barriers. This study examines the population level genetic divergence and phylogenetic relationships of species in the shortfin group of the subgenus Mollienesia (genus Poecilia), a group of live-bearing fishes that are widely distributed across Mexico, with sampling at a small geographic scale. Samples from over 50 locations were analyzed for six species by using phylogenetic and haplotype network approaches to assess genetic diversity across geographic ranges and to refine the distributions of species in this group. The results indicate that Mexican species have diversified following multiple, independent invasions from Middle America. Two species found north of the Trans-Mexican Volcanic Belt (TMVB) and one transversal species exhibited weak phylogenetic structure, likely due to the lack of physiographic barriers, recent colonization, and high dispersal rates among regions. In contrast, three species found south of the TMVB exhibited strong phylogenetic structure, reflecting a longer presence in the area and multiple physiographic barriers that isolated populations. This study identified mechanisms driving divergence and speciation, expanded the known range of several species, and resolved taxonomic uncertainties of populations. [ABSTRACT FROM AUTHOR]

Velifera molly (Poecilia velifera) has an ecophysiological interest related to euryhalinity. To evaluate its tolerance and performance in different salinities, and the ability to be reared at diverse locations, Median Lethal Salinity (LS50) and growth performance were investigated. In experiment 1, 120 juveniles were tested through direct transfer to salinities 0, 24, 48, 60, 72, and 78 psu for 96 h. In experiment 2, 160 juveniles were gradually acclimatized to salinities 0, 12, 24, and 36 psu. The experiment was conducted using recirculation aquaculture systems (RAS) for 75 d. LS50-96 h was estimated at 71.1 psu and safe salinity at 7.1 psu. The final weight, weight gain, and feed intake had an inverse relationship to salinity, reflecting the sublethal effects. The apparent feed conversion rate was directly related to salinity, presented a better performance in freshwater conditions. There was no mortality in experiment 2, allowing the species to be maintained for long periods in the analyzed salinities. [ABSTRACT FROM AUTHOR]

The sensory drive hypothesis predicts that across different light environments sexually selected colour patterns will change to increase an animal's visual communication efficiency within different habitats. This is because individuals with more efficient signal components are likely to have more successful matings and hence produce more offspring. However, how colour pattern signals change over multiple generations under different light environmental conditions has not been tested experimentally. Here, we manipulated colour pattern signal efficiency by providing different ambient light environments over multiple generations to examine whether male colour pattern components change within large replicated populations of guppies (Poecilia reticulata). We report that colour patches change within populations over time and are phenotypically different among our three different light environments. Visual modelling suggests that the majority of these changes can be understood by considering the chroma, hue and luminance of each colour patch as seen by female guppies under each light environment. Taken together, our results support the hypothesis that different environmental conditions during signal reception can directly or indirectly drive the phenotypic diversification of visual signals within species.

Sensory drive proposes that natural selection on nonmating behaviours (e.g. foraging preferences) alters sensory system properties and results in a correlated effect on mating preferences and subsequently sexual traits. In colour-based systems, we can test this by selecting on nonmating colour preferences and testing for responses in colour-based female preferences and male sexual coloration. In guppies (Poecilia reticulata), individual functional links of sensory drive have been demonstrated providing an opportunity to test the process over more than one link. We measured male coloration and female preferences in populations previously artificially selected for colour-based foraging behaviour towards two colours, red and blue. We found associated changes in male coloration in the expected direction as well as weak changes in female preferences. Our results can be explained by a correlated response in female preferences due to artificial selection on foraging preferences that are mediated by a shared sensory system or by other mechanisms such as colour avoidance, pleiotropy or social experiences. This is the first experimental evidence that selection on a nonmating behaviour can affect male coloration and, more weakly, female preferences.

The degree of divergence between the sex chromosomes is not always proportional to their age. In poeciliids, four closely related species all exhibit a male heterogametic sex chromosome system on the same linkage group, yet show a remarkable diversity in X and Y divergence. In Poecilia reticulata and P. wingei , the sex chromosomes remain homomorphic, yet P. picta and P. parae have a highly degraded Y chromosome. To test alternative theories about the origin of their sex chromosomes, we used a combination of pedigrees and RNA-seq data from P. picta families in conjunction with DNA-seq data collected from P. reticulata , P. wingei , P. parae , and P. picta. Phylogenetic clustering analysis of X and Y orthologs, identified through segregation patterns, and their orthologous sequences in closely related species demonstrates a similar time of origin for both the P. picta and P. reticulata sex chromosomes. We next used k-mer analysis to identify shared ancestral Y sequence across all four species, suggesting a single origin to the sex chromosome system in this group. Together, our results provide key insights into the origin and evolution of the poeciliid Y chromosome and illustrate that the rate of sex chromosome divergence is often highly heterogenous, even over relatively short evolutionary time frames. [ABSTRACT FROM AUTHOR]

Objective: Some cationic anti-microbial peptides show a wide range of cytotoxic action versus malignant cells, which may lead to developing a novel group of antitumor medications. In the present study, the anticancer activity of pleurocidin-like peptide WF3 isoform X2 (AMP-WF3), from the Poecilia Mexicana fish, against leukemic cell line Jurkat was evaluated, and the cytotoxicity compared with the effects on normal cells, including peripheral blood mononuclear cells (PBMCs) and human dermal fibroblast (HDF) cells. Materials and Methods: In this experimental study, cells were treated with various dosages of AMP-WF3 for 24 hours. Using methyl thiazole tetrazolium salt reduction (MTT test), the effects of the AMP-WF3 on cell viability and toxicity were evaluated. The impact of this peptide on apoptotic pathways was examined using flow cytometry and Annexin V-PI stains. Additionally, the relative expression of the P53, P21, and BCL-2 genes was evaluated using a real-time polymerase chain reaction. Results: The Jurkat cell line was more susceptible to AMP-WF3 cytotoxicity [half-maximal inhibitory concentration (IC50)=50 μM], while normal cells (PBMCs and HDF) were less susceptible. Flow cytometry verified that the apoptotic activity of AMP-WF3 on Jurkat cells was significantly higher than that of HDF and PBMCs. Peptide-treated Jurkat cells were associated with increased expression of P21, and P53 genes. In contrast, the changes in P21, P53, and BCL-2 genes differed in PBMCs and HDF cells. In HDF cells, simultaneous increase of P21, P53, and BCL-2, and in PBMCs, only the increase of BCL-2 was observed. Conclusion: Our research showed that AMP-WF3 could be developed as a novel treatment agent with minimum side effects for ALL patients. [ABSTRACT FROM AUTHOR]

Members of the fish family Poeciliidae (livebearing 'tooth-carps') have historically been used as models in medical research, behavior ecology, and biological control. This group of primarily freshwater fishes is highly tolerant to environmental factors such as salinity and warm temperatures and includes some invasive species. Here, we present the mitochondrial genome of Poecilia parae. A representative of this species was obtained from Suriname. The complete mitochondrial genome was sequenced using Oxford Nanopore technology and is 16,559 bp long. The genome contains 13 protein-coding genes, two ribosomal RNAs (rRNAs), 22 transfer RNAs (tRNAs), and one control region (D-loop). Phylogenetic analysis yielded topologies similar to those previously published. The data generated here will be useful in future studies of comparative biology and those utilizing environmental DNA (eDNA). [ABSTRACT FROM AUTHOR]

The present study examined the effects of environmentally realistic exposure to atrazine (ATZ) on the behavior of sailfin mollies, Poecilia latipinna. ATZ is one of the most commonly used pesticides in the US and a known endocrine disrupting chemical (EDC). The behavior of sailfin mollies is well documented in the scientific literature. Moreover, they are ecologically important indicators of environmental health, widely distributed among the mangroves along the Gulf Coast of the Southeastern US where significant amounts of ATZ are introduced via runoff and storm water drainage from coastal households, businesses, and farms. Four sets of experiments designed to assess ATZ's impact on various aspects of male and female reproductive behavior, aggression, anxiety, and boldness were conducted following 12 weeks of exposure to 1 or 15 ppb water-borne ATZ, along with a no-exposure control group. Results indicated that the behavior of ATZ-exposed individuals differed from those of controls: ATZ exposure affected which stimulus fish (a male vs. a female) subject females preferred to associate with while also affecting female strength of preference for males of larger body size and their sexual receptivity to conspecific males in general. ATZ-exposed males also showed reduced overall responsiveness to conspecific stimuli and directed significantly less aggression toward their mirror image compared with controls. Finally, ATZ exposure affected multiple aspects of male and female behavior that are often used as proxies for boldness and anxiety. Overall, ATZ exposure resulted in alterations across a variety of behaviors attributed to sexual receptivity, mate choice and motivation to mate, aggression, as well as boldness and anxiety. These ATZ-induced behavioral changes may adversely affect the long-term health of natural populations exposed to similar, environmentally realistic concentrations and add to a growing body of empirical data demonstrating substantial fitness consequences of exposure to sublethal concentrations of this known EDC. [ABSTRACT FROM AUTHOR]

Poecilia parae, or penta, stands out for its vibrant colors, livebearing nature, and ecological importance. Penta is important for understanding ecology, evolution, and behavior due to its adaptability, reproductive strategies, ecological role, genetic diversity, and relevance to conservation efforts. The knowledge gained from studying this species contributes to a broader understanding of freshwater ecosystems and the factors influencing the dynamics of aquatic communities. Understanding the diversity of body colors and morphs in P. parae provides valuable insights into the species' ecology, evolutionary history, and adaptation strategies. Additionally, the visual appeal of these fish makes them popular choices for aquarium enthusiasts, leading to further exploration of selective breeding and the development of distinct color variants in captivity. This paper presents the alternative of using a new poeciliid, in addition to the well-known guppy, for behavioral studies, behavioral ecology and evolutionary ecology. [ABSTRACT FROM AUTHOR]

Iron oxide nanoparticles (IONPs) are being increasingly recognized as viable materials for environmental remediation due to their capacity to adsorb contaminants such as glyphosate (GLY) on their surfaces. Nevertheless, the ecotoxicological implications of IONPs associated with GLY necessitate thorough evaluation to ascertain the safety of such remediation strategies. In this context, the present investigation was conducted to examine hepatic biomarkers pertinent to the redox system, as well as ultrastructural hepatic alterations in Poecilia reticulata, following a 21-day exposure to environmentally relevant concentrations of IONPs, iron ions (Fe), and glyphosate in its pure form (GLY) as well as a commercial glyphosate-based herbicide (GBH). After this exposure, the fish underwent a 21-day recovery in uncontaminated water. The results indicated an increase in the activity of catalase (CAT) and glutathione S-transferase (GST) and in the concentration of glutathione (GSH) in the animals subjected to IONP+GBH and IONP+GLY treatments. This biochemical response persisted for the duration of both the exposure and recovery phases. Concurrently, hepatocytes displayed mitochondria with increased electron density, augmented lipid droplet accumulation, and expanded necrotic areas within the hepatic tissue. In contrast, fish exposed solely to IONPs exhibited sustained redox homeostasis throughout the investigative timeline. These findings suggest that the coexposure toxicity of IONP+GLY and IONP+GBH is attributable to the agent adsorbed onto the IONPs and that P. reticulata could maintain an active antioxidant defense mechanism throughout the entire study period., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Studies of sex chromosome systems at early stages of divergence are key to understanding the initial process and underlying causes of recombination suppression. However, identifying signatures of divergence in homomorphic sex chromosomes can be challenging due to high levels of sequence similarity between the X and the Y. Variations in methodological precision and underlying data can make all the difference between detecting subtle divergence patterns or missing them entirely. Recent efforts to test for X‐Y sequence differentiation in the guppy have led to contradictory results. Here, we apply different analytical methodologies to the same data set to test for the accuracy of different approaches in identifying patterns of sex chromosome divergence in the guppy. Our comparative analysis reveals that the most substantial source of variation in the results of the different analyses lies in the reference genome used. Analyses using custom‐made genome assemblies for the focal population or species successfully recover a signal of divergence across different methodological approaches. By contrast, using the distantly related Xiphophorus reference genome results in variable patterns, due to both sequence evolution and structural variations on the sex chromosomes between the guppy and Xiphophorus. Changes in mapping and filtering parameters can additionally introduce noise and obscure the signal. Our results illustrate how analytical differences can alter perceived results and we highlight best practices for the study of nascent sex chromosomes. [ABSTRACT FROM AUTHOR]

The impacts of relying on stored sperm were evaluated in the sailfin molly, Poecilia latipinna. Females reliant on stored sperm had fewer offspring compared to remated females, but offspring size and short‐term growth rate did not differ. Thus, females may use stored sperm in cases such as previous mating with a preferred male, lack of access to mating opportunities during a reproductive cycle, or to maximize egg fertilization. Females do not compensate for producing fewer offspring however, by allocating more resources to offspring relative to their size or initial growth. [ABSTRACT FROM AUTHOR]

Addition of canals and levees to wetlands is common in hydrological management. Permanently flooded canals provide fishes with refuge from desiccation and corridors for long-distance movement, but also may present high risk of predation. Levees create barriers to movement. We evaluated the effect of canals and levees on the movement of fish in seasonally fluctuating marshes in Everglades National Park between 2003 and 2016. We used directional traps to quantify activity and directional movement of seven species of fishes moving through marshes near canals and levees, and farther into the wetland. Eastern mosquitofish (Gambusia holbrooki) were ubiquitous, were active at all sites throughout the year, and moved towards canals consistent with their use as refuge habitat. Sailfin mollies (Poecilia latipinna) and bluefin killifish (Lucania goodei) were also likely to swim towards canals as water levels dropped in the dry season. Flagfish (Jordanella floridae), golden topminnows (Fundulus chrysotus), and marsh killifish (F. confluentus) used wetlands associated with anthropogenic structures but showed no directional bias with respect to canals and these species did not appear to use them as a drought refuge. Non-native African Jewelfish (Hemichromis letourneauxi) displayed directed movement related to canals, but were most active at sites distant from canals. This study provides evidence for interspecific differences among seven fishes in how they move through a marsh near different anthropogenic structures and in different seasons of the hydrologic year. Modification of wetland landscapes with structures has implications for species sorting and metacommunity dynamics filtered by species-specific behavioral traits. Monitoring efforts like this support understanding how important members of the community, like small fishes, respond to environmental and anthropogenic factors that are subject to management decisions. [ABSTRACT FROM AUTHOR]

The colonization of novel environments often involves changes in gene expression, protein coding sequence, or both. Studies of how populations adapt to novel conditions, however, often focus on only one of these two processes, potentially missing out on the relative importance of different parts of the evolutionary process. In this study, our objectives were 1) to better understand the qualitative concordance between conclusions drawn from analyses of differential expression and changes in genic sequence and 2) to quantitatively test whether differentially expressed genes were enriched for sites putatively under positive selection within gene regions. To achieve this, we compared populations of fish (Poecilia mexicana) that have independently adapted to hydrogen-sulfide-rich environments in southern Mexico to adjacent populations residing in nonsulfidic waters. Specifically, we used RNA-sequencing data to compare both gene expression and DNA sequence differences between populations. Analyzing these two different data types led to similar conclusions about which biochemical pathways (sulfide detoxification and cellular respiration) were involved in adaptation to sulfidic environments. Additionally, we found a greater overlap between genes putatively under selection and differentially expressed genes than expected by chance. We conclude that considering both differential expression and changes in DNA sequence led to a more comprehensive understanding of how these populations adapted to extreme environmental conditions. Our results imply that changes in both gene expression and DNA sequence-sometimes at the same loci-may be involved in adaptation.

This study aims to propose new fish model organisms for experimental studies on anisakis and other fish pathogens as alternatives to the commonly used fish model Danio rerio (Hamilton, 1822). For this purpose, the sailfin molly (Poecilia latipinna (Lesueur, 1821)) and the gilthead seabream (Sparus aurata Linnaeus, 1758) were tested by experimental infections with Anisakis simplex s.l. larvae. Preliminary susceptibility challenges on mollies showed successful infections, with larvae observed up to 384 h post-infection (hpi). Histological analysis showed encapsulation of larvae and mild tissue damage in the infected fish, providing insight into the infection process. Two infection methods, oral ingestion and orogastric inoculation, were tested in both fish species. The results indicate successful infections with both methods, with higher prevalence and recovery rates observed with orogastric inoculation (especially in gilthead seabream). In addition, the study examined the chronology of the infective process, histological changes, and encapsulation patterns in infected fish. This comprehensive study highlights the suitability of sailfin molly as a model organism for experimental studies on anisakid infections and provides valuable insights into infection dynamics and host-parasite interactions. Further research in this area could contribute to the development of effective prevention and control measures for anisakid infections, and thus reduce the health risks associated with fish consumption. • Sparus aurata and Poecilia latipinna, are susceptible to Anisakis spp. infections. • Anisakis infection by orogastric inoculation was more successful than oral ingestion. • Mollies are proposed as valuable model organisms for experimental infections. [ABSTRACT FROM AUTHOR]

Variation in gene expression can provide insights into organismal responses to environmental stress and physiological mechanisms mediating adaptation to habitats with contrasting environmental conditions. We performed an RNA-sequencing experiment to quantify gene expression patterns in fish adapted to habitats with different combinations of environmental stressors, including the presence of toxic hydrogen sulphide (H2 S) and the absence of light in caves. We specifically asked how gene expression varies among populations living in different habitats, whether population differences were consistent among organs, and whether there is evidence for shared expression responses in populations exposed to the same stressors. We analysed organ-specific transcriptome-wide data from four ecotypes of Poecilia mexicana (nonsulphidic surface, sulphidic surface, nonsulphidic cave and sulphidic cave). The majority of variation in gene expression was correlated with organ type, and the presence of specific environmental stressors elicited unique expression differences among organs. Shared patterns of gene expression between populations exposed to the same environmental stressors increased with levels of organismal organization (from transcript to gene to physiological pathway). In addition, shared patterns of gene expression were more common between populations from sulphidic than populations from cave habitats, potentially indicating that physiochemical stressors with clear biochemical consequences can constrain the diversity of adaptive solutions that mitigate their adverse effects. Overall, our analyses provided insights into transcriptional variation in a unique system, in which adaptation to H2 S and darkness coincide. Functional annotations of differentially expressed genes provide a springboard for investigating physiological mechanisms putatively underlying adaptation to extreme environments.

Population variation in trophic niche is widespread among organisms and is of increasing interest given its role in both speciation and adaptation to changing environments. Trinidadian guppies (Poecilia reticulata) inhabiting stream reaches with different predation regimes have rapidly evolved divergent life history traits. Here, we investigated the effects of both predation and resource availability on guppy trophic niches by evaluating their gut contents, resource standing stocks, and δ15N and δ13C stable isotopes across five streams during the wet season. We found that guppies from low predation (LP) sites had a consistently higher trophic position and proportion of invertebrates in their guts and assimilate less epilithon than guppies from high predation (HP) sites. Higher trophic position was also associated with lower benthic invertebrate availability. Our results suggest that LP guppies could be more efficient invertebrate consumers, possibly as an evolutionary response to greater intraspecific competition for higher quality food. This may be intensified by seasonality, as wet season conditions can alter resource availability, feeding rates, and the intensity of intraspecific competition. Understanding how guppy diets vary among communities is critical to elucidating the role of niche shifts in mediating the link between environmental change and the evolution of life histories.

Behavioural individuality is thought to be caused by differences in genes and/or environmental conditions. Therefore, if these sources of variation are removed, individuals are predicted to develop similar phenotypes lacking repeatable individual variation. Moreover, even among genetically identical individuals, direct social interactions are predicted to be a powerful factor shaping the development of individuality. We use tightly controlled ontogenetic experiments with clonal fish, the Amazon molly (Poecilia formosa), to test whether near-identical rearing conditions and lack of social contact dampen individuality. In sharp contrast to our predictions, we find that (i) substantial individual variation in behaviour emerges among genetically identical individuals isolated directly after birth into highly standardized environments and (ii) increasing levels of social experience during ontogeny do not affect levels of individual behavioural variation. In contrast to the current research paradigm, which focuses on genes and/or environmental drivers, our findings suggest that individuality might be an inevitable and potentially unpredictable outcome of development.