Cortisol enhances aerobic metabolism and locomotor performance during the transition to land in an amphibious fish.

Amphibious fishes on land encounter higher oxygen (O2) availability and novel energetic demands, which impacts metabolism. Previous work on the amphibious mangrove killifish (Kryptolebias marmoratus) has shown that cortisol becomes elevated in response to air exposure, suggesting a possible role in regulating metabolism as fish move into terrestrial environments. We tested the hypothesis that cortisol is the mechanism by which oxidative processes are upregulated during the transition to land in amphibious fishes. We used two groups of fish, treated fish (+metyrapone, a cortisol synthesis inhibitor) and control (-metyrapone), to determine the impact of cortisol during air exposure (0 and 1 h, 7 days) on O2 consumption, terrestrial locomotion, the phenotype of red skeletal muscle, and muscle lipid concentration. Metyrapone-treated fish had an attenuated elevation in O2 consumption rate during the water to air transition and an immediate reduction in terrestrial exercise performance relative to control fish. In contrast, we found no short- (0 h) or long-term (7 days) differences between treatments in the oxidative phenotype of red muscles, nor in muscle lipid concentrations. Our results suggest that cortisol stimulates the necessary increase in aerobic metabolism needed to fuel the physiological changes that amphibious fishes undergo during the acclimation to air, although further studies are required to determine specific mechanisms of cortisol regulation.

Nanosilica and copper ecotoxicity in Gambusia holbrooki fish.

When silica nanoparticles (SiNP) reach the water bodies interact with the already existing pollutants in the environments. This study aimed to evaluate the ecotoxicity of SiNP under the presence/absence of Cu in mosquitofish (Gambusia holbrooki). Fish were exposed to 0, 10 and 100 mg SiNP L-1, alone or mixed with Cu (0.25 mg L-1). After 96 h, the amount of colony forming units (CFU) of bacteria living on the skin mucus was analysed, and oxidative stress, tissue damage enzymes, and neurotoxicity were evaluated. We observed a reduction in CFU when Cu was present in the media. The liver was the target organ, evidencing a decrease in tissue damage enzymatic activities, activation of the antioxidant system in all treatments, and lipid oxidative damage when the SiNP and Cu were mixed. Overall, SiNP ecotoxicity was proved, which could also be enhanced by the presence of ubiquitous elements such as metals.

Social experience influences thermal sensitivity: lessons from an amphibious mangrove fish.

Understanding the factors affecting the capacity of ectothermic fishes to cope with warming temperature is critical given predicted climate change scenarios. We know that a fish's social environment introduces plasticity in how it responds to high temperature. However, the magnitude of this plasticity and the mechanisms underlying socially modulated thermal responses are unknown. Using the amphibious hermaphroditic mangrove rivulus fish Kryptolebias marmoratus as a model, we tested three hypotheses: (1) social stimulation affects physiological and behavioural thermal responses of isogenic lineages of fish; (2) social experience and acute social stimulation result in distinct physiological and behavioural responses; and (3) a desensitization of thermal receptors is responsible for socially modulated thermal responses. To test the first two hypotheses, we measured the temperature at which fish emerged from the water (i.e. pejus temperature) upon acute warming with socially naive isolated fish and with fish that were raised alone and then given a short social experience prior to exposure to increasing temperature (i.e. socially experienced fish). Our results did not support our first hypothesis as fish socially stimulated by mirrors during warming (i.e. acute social stimulation) emerged at similar temperatures to isolated fish. However, in support of our second hypothesis, a short period of prior social experience resulted in fish emerging at a higher temperature than socially naive fish suggesting an increase in pejus temperature with social experience. To test our third hypothesis, we exposed fish that had been allowed a brief social interaction and naive fish to capsaicin, an agonist of TRPV1 thermal receptors. Socially experienced fish emerged at significantly higher capsaicin concentrations than socially naive fish suggesting a desensitization of their TRPV1 thermal receptors. Collectively, our data indicate that past and present social experiences impact the behavioural response of fish to high temperature. We also provide novel data suggesting that brief periods of social experience affect the capacity of fish to perceive warm temperature.

An effect assessment of microplastics and nanoplastics interacting with androstenedione on mosquitofish (Gambusia affinis).

An increasing number of microplastics have been detected in aquatic environments, causing various damage to organisms. The size of microplastics affects the toxicity once they enter the organisms. Meanwhile, there is an increasing variety of Endocrine-disrupting chemicals (EDCs) present in aquatic environments. Androstenedione (AED) is a typical EDC. In this study, we used polystyrene microspheres of 80 nm (NPs) and 8 µm (MPs) as materials to simulate environmental contaminants in the aquatic environment with AED. We used female mosquitofish (Gambusia affinis) as the research object to investigate the effects of microplastics on fish in waters containing AED. We compared different sizes of particles accumulation in some tissues of fish and variation of enzyme activities (SOD, LDH, CAT), and the content of MDA in the gut. MPs, NPs, and AED combined exposure test investigated mRNA profiles of immune-related genes (IL-1ß, IL-6, IL-8, IL-10) and hormone receptor genes (ARα, ARß, ERα, ERß) in the liver of fish. Our results indicated that MPs emerged in various tissues (gill, gut, and liver) of mosquitofish. Besides, NPs and MPs caused enteric abnormal enzyme activity after 48 h of exposure, which was particularly pronounced in the MPs-AED group. MPs induced significant upregulation of inflammatory factors and gonadal factor genes after 96 h of exposure, which was more pronounced when co-exposed with AED. In conclusion, NPs and MPs caused mechanisms of immune damage and inflammatory response. MPs were found to be more likely to cause adverse reactions than NPs, and these responses were enhanced by the combined effects of AED. This study demonstrated that AED can exacerbate the negative effects of MPs and NPs on mosquitofish. It provided an important basis for the effective assessment of MPs and NPs on bioaccumulation and biochemical status of mosquitofish. Additionally, it serves as a foundation to investigate the interactive effects of microplastics and EDCs in living organisms.

Widespread psychoactive pollutant augments daytime restfulness and disrupts diurnal activity rhythms in fish.

Pharmaceutical pollution is a major driver of global change, with the capacity to alter key behavioural and physiological traits in exposed animals. Antidepressants are among the most commonly detected pharmaceuticals in the environment. Despite well-documented pharmacological effects of antidepressants on sleep in humans and other vertebrates, very little is known about their ecologically relevant impacts as pollutants on non-target wildlife. Accordingly, we investigated the effects of acute 3-day exposure of eastern mosquitofish (Gambusia holbrooki) to field-realistic levels (nominal concentrations: 30 and 300 ng/L) of the widespread psychoactive pollutant, fluoxetine, on diurnal activity patterns and restfulness, as indicators of disruptions to sleep. We show that exposure to fluoxetine disrupted diel activity patterns, which was driven by augmentation of daytime inactivity. Specifically, unexposed control fish were markedly diurnal, swimming farther during the day and exhibiting longer periods and more bouts of inactivity at night. However, in fluoxetine-exposed fish, this natural diel rhythm was eroded, with no differences in activity or restfulness observed between the day and night. As a misalignment in the circadian rhythm has been shown to adversely affect fecundity and lifespan in animals, our findings reveal a potentially serious threat to the survival and reproductive success of pollutant-exposed wildlife.

Whole-Brain Clearing and Immunofluorescence in Nothobranchius furzeri.

Tissue clearing techniques for three-dimensional reconstruction and imaging of entire organs and thick samples have become a popular and broadly used methodology, leading to the development of numerous protocols. Due to the complex cellular architecture of the brain and the wide spatial range of the connections that neurons may display, having the possibility to stain, image, and reconstruct neurons and/or neuronal nuclei in their entire extent can be crucial. However, this is hard to accomplish due to the natural opacity of the brain and the general thickness of the sample, posing a barrier to both imaging and antibody penetration. Nothobranchius furzeri has recently become a widely used model to study brain aging thanks to its short life span (3-7 mo), providing new opportunities to study the effects of aging on the brain and the role of aging in the development of neurodegenerative diseases. Here, we present a methodology to clarify and stain whole N. furzeri brains. This protocol is based on the ScaleA2 and ScaleS protocols developed and presented by Hama and colleagues, together with an in-house developed staining procedure for thick slices of tissues. ScaleS is a convenient and easy clearing technique based on sorbitol and urea that does not require particularly complex equipment, but due to the high urea concentration in some of the solutions, not all antigens are preserved. To overcome this issue, we developed a method that allows optimal staining of Nothobranchius furzeri brains before clarification.

Predation history has no effect on lateralized behavior in Brachyrhaphis rhabdophora.

Evolutionary biologists have grown increasingly interested in laterality, a phenomenon where bilaterally symmetrical organisms show a side bias in some trait. Lateralized behavior is particularly interesting because it is not necessarily tied to morphological asymmetry. What causes lateralized behavior remains largely unknown, although previous research in fishes suggest that fish might favor one eye over another to view potential food sources, mates, and to assess predation risk. Here we test the hypothesis that a history of predation risk predicts lateralized behavior in the livebearing fish Brachyrhaphis rhabdophora. To do this, we used a detour assay to test for eye bias when a focal fish approached various stimuli (predator, potential mate, novel object, and empty tank control). Contrary to our predictions, we found no differences in lateralized behavior between fish from populations that co-occurred with fish predators relative to those that do not co-occur with predators. In fact, we found no evidence for behavioral lateralization at all in response to any of the stimuli. We explore several possible explanations for why lateralized behavior is absent in this species, especially considering a large body of work in other livebearing fishes that shows that lateralized behavior does occur.

Nonlethal Blood Sampling from the Killifish Nothobranchius furzeri.

Blood withdrawal is a common procedure performed on laboratory animals to monitor key processes and indicators of fish health and physiology, such as hematopoiesis, hemostasis, and lipid and glucose metabolism. Moreover, the ability to extract blood with minimal invasiveness and without sacrificing animals enables repeated sampling, allowing both longitudinal studies of individual animals, as well as reducing the number of experimental animals needed in a study. The African turquoise killifish is an emerging animal model that is progressively being adopted worldwide for aging studies because of its naturally short life span. However, because of the small body size of this species, nonlethal blood collection is a challenging procedure. Here we present a detailed protocol enabling repeated blood sampling from the same individual fish. This method, if correctly executed, is minimally invasive and does not cause any lasting damage. The protocol has been tested on animals spanning from 6 to 24 wk of age and the amount of blood that could be extracted varied from 0.5 to 8 µL, greatly depending on specimen age, sex, and size. This volume is sufficient to perform analyses such as blood glucose measurement, blood cell counts, or histological stains on blood smears.

Functional and Behavioral Trade-Offs between Terrestrial and Aquatic Locomotion in the Amphibious Fish Kryptolebias marmoratus.

The mangrove rivulus (Kryptolebias marmoratus) is a phenotypically plastic teleost fish that can spend considerable time on land and traverse the terrestrial realm through a behavior termed the tail-flip jump. The tail-flip jump is a transitional stage between fully aquatic and terrestrial lifestyles. Therefore, understanding this behavior can provide insight into how organisms adapt to new environments over evolutionary time. Studies of K. marmoratus show that terrestrial acclimation and exercise improve tail-flip jumping performance due to muscle remodeling, but the implications of these muscular changes on aquatic locomotion are unknown. In the present study, we hypothesized that (1) terrestrial acclimation and exercise lead to physiological changes, such as changes to muscle fiber type, muscle mass distribution, or body shape, that optimize tail-flip jump distance and endurance while negatively impacting swimming performance in K. marmoratus, and (2) plasticity of the brain (which has been demonstrated in response to a variety of stimuli in K. marmoratus) allows terrestrial emersion and exercise to cause behavioral changes that promote survival and long-term reproductive success. To test these hypotheses, we measured the critical swimming speed (Ucrit), tail-flip jump distance, terrestrial endurance, and undisturbed aquatic behavior of age- and size-matched K. marmoratus before and after a terrestrial exercise period. This period consisted of six 3-min exercise sessions spread over 12 days, during which the fish were prompted to jump continuously. To isolate the effects of air exposure, a separate group was exposed to air for an equivalent period but not allowed to jump. Air exposure improved maximum jump distance but negatively affected swimming performance (Ucrit). Terrestrial endurance (number of jumps) improved in the exercised group, but Ucrit showed no significant change. Contrary to our first hypothesis, a trade-off exists between jump distance and Ucrit but not between jump endurance and Ucrit. Exercised individuals were more active following exercise, resulting either from the onset of dispersion behavior or a heightened stress response.

Thyroid endocrine disruption and neurotoxicity of gestodene in adult female mosquitofish (Gambusia affinis).

The frequent detection of progestins in various aquatic environments and their potential endocrine disruptive effects in fish have attracted increasing attention worldwide. However, data on their effects on thyroid function and neurotoxicity in fish are limited, and the underlying mechanisms remain unclear. Here, the effects of gestodene (GES, a common progestin) on the thyroid endocrine and nervous systems of mosquitofish (Gambusia affinis) were studied. Adult female fish were exposed to GES at environmentally relevant concentrations (4.4-378.7 ng/L) for 60 days. The results showed that exposure to 378.7 ng/L GES caused a significant decrease in fish growth compared with the control and a marked reduction in the total distance traveled (50.6%) and swimming velocity (40.1-61.9%). The triiodothyronine (T3) levels were significantly increased by GES in a dose-dependent manner, whereas those of tetraiodothyronine (T4) were significantly decreased only at the G500 concentration. The acetylcholinesterase (AChE) activity was decreased significantly in the 4.42 ng/L GES treatments, but increased significantly at 378.67 ng/L. In the brain, a strong increase in the transcriptional levels of bdnf, trh, and dio2 was observed in fish after the 378.7 ng/L treatment. In addition, chronic exposure to GES caused colloid depletion with a concentration-dependent manner in the thyroid, and angiectasis, congestion, and vacuolar necrosis in the brain. These findings provide a better understanding of the effects of GES and associated underlying mechanisms in G. affinis.

Increased superfetation precedes the evolution of advanced degrees of placentotrophy in viviparous fishes of the family Poeciliidae.

The causes and consequences of the evolution of placentotrophy (post-fertilization nutrition of developing embryos of viviparous organisms by means of a maternal placenta) in non-mammalian vertebrates are still not fully understood. In particular, in the fish family Poeciliidae there is an evolutionary link between placentotrophy and superfetation (ability of females to simultaneously bear embryos at distinct developmental stages), with no conclusive evidence for which of these two traits facilitates the evolution of more advanced degrees of the other. Using a robust phylogenetic comparative method based on Ornstein-Uhlenbeck models of adaptive evolution and data from 36 poeciliid species, we detected a clear causality pattern. The evolution of extensive placentotrophy has been facilitated by the preceding evolution of more simultaneous broods. Therefore, placentas became increasingly complex as an adaptive response to evolutionary increases in the degree of superfetation. This finding represents a substantial contribution to our knowledge of the factors that have shaped placental evolution in poeciliid fishes.

Are superficial neuromasts proprioceptors underlying fast copulatory behavior?

In male Poeciliid fishes, the modified anal fin (i.e., gonopodium) and its axial and appendicular support are repositioned within the axial skeleton, creating a novel sexually dimorphic ano-urogenital region. During copulation, the relative location of the gonopodium is crucial for successful insemination. Therefore, the repositioning of these structures and organ relied on the reorganization of the efferent circuitry that controls spinal motor neurons innervating appendicular muscles critical for the movement of the gonopodium, including the fast and synchronous torque-trust motion during insemination attempts. Copulation occurs when a male positions himself largely outside a female's field of view, circumducts his gonopodium, and performs a rapid, complex maneuver to properly contact the female urogenital sinus with the distal tip of the gonopodium and transfers sperm. Although understanding of the efferent circuitry has significantly increased in the last 24 years, nothing is known about the cutaneous receptors involved in gonopodium movement, or how the afferent signals are processed to determine the location of this organ during copulation. Using Western mosquitofish, Gambusia affinis, as our model, we attempt to fill this gap in knowledge. Preliminary data showed cutaneous nerves and sensory neurons innervating superficial neuromasts surrounding the base of adult male gonopodium; those cutaneous nerves projected ventrally from the spinal cord through the 14th dorsal root ganglion and its corresponding ventral root towards the base and fin rays of the gonopodium. We asked what role the cutaneous superficial neuromasts play in controlling the positioning and timing of the gonopodium's fast and synchronous movements for effective sperm transfer. First, we found a greater number of superficial neuromasts surrounding the base of the male's gonopodium compared to the base of the female's anal fin. Second, we systemically removed superficial neuromasts surrounding the gonopodium base and observed significant impairment of the positioning and timing of gonopodial movements. Our findings provide a first step to supporting the following hypothesis: during radical reorganization of the Poeciliid body plan, superficial neuromasts have been partially co-opted as proprioceptors that allow the gonopodium to control precise positioning and timing during copulatory attempts.

Administration of krill oil extends lifespan of fish Nothobranchius guentheri via enhancement of antioxidant system and suppression of NF-κB pathway.

Krill oil (KO) extracted from Antarctic krill (Euphausia superba) mainly comprises phospholipids and triglycerides. KO has been shown to prolong the median lifespan of the nematode Caenorhabditis elegans, but to shorten the lifespan of long-lived F1 mice; therefore, it remains controversial over the life-extending property of KO. In this study, we clearly demonstrated that dietary intake of KO extended both the mean and maximum lifespans of aged male Nothobranchius guentheri (p < 0.05), reduced the accumulation of lipofuscin (LF) (p < 0.05) in the gills and senescence-associated ß-galactosidase (SA-ß-Gal) (p < 0.05) in the caudal fins, and lowered the levels of protein oxidation (p < 0.05), lipid peroxidation (p < 0.01), and reactive oxygen species (ROS) (p < 0.01) in the muscles and livers, indicating that KO possesses rejuvenation and anti-aging activity. We also showed that KO enhanced the activities of antioxidant enzymes catalase (CAT) (p < 0.05), superoxide dismutase (SOD) (p < 0.05), and glutathione peroxidase (GPX) (p < 0.05) in aged male N. guentheri. In addition, KO administration effectively reversed histological lesions including inflammatory cell infiltration and structural collapse in the muscles and livers of aged N. guentheri and suppressed the nuclear factor kappa-B (NF-κB) signaling pathway (p < 0.05), a master regulator of inflammation. Altogether, our study indicates that KO has anti-aging and rejuvenation property. It also suggests that KO exerts its anti-aging and rejuvenation effects via enhancement of the antioxidant system and suppression of the NF-κB signaling pathway.

How maternal age and environmental cues influence embryonic developmental pathways and diapause dynamics in a North American annual killifish.

BACKGROUND: Temporary pools are variable environments with seasonal drought/flood phases. Annual killifish have adapted to life in temporary pools by producing embryos that undergo diapause to traverse the dry phase. To fill existing knowledge gaps about embryo diapause regulation and evolution in annual killifishes, we test the effect of maternal age, incubation temperature, and incubation medium on diapause induction and length in Millerichthys robustus, the only North American fish species that has evolved an annual life history. RESULTS: All embryos at extreme temperatures follow a defined developmental pathway: skipping diapause at 30°C, and entering diapause at 18°C, both regardless of maternal age, and incubation medium. However, maternal age, and incubation medium influenced whether diapause is entered, and time arrested in diapause for embryos incubated at 25°C. At 25°C, five-week-old, and 52-week-old females produced more embryos that entered diapause than 26-week-old females. Also, embryos incubated in aqueous medium skipped diapause more frequently at this intermediate temperature. CONCLUSIONS: Millerichthys developmental dynamics associated with maternal age under intermediate range of temperatures are likely adapted to the particular patterns of flood/drought in North American temporary pools. Millerichthys also exhibits developmental patterns largely comparable with other annual fishes, probably due to common seasonal patterns in temporary pools.

Increasing salinization of freshwater limits invasiveness of a live-bearing fish: Insights from behavioral and life-history traits.

Biological invasions and continued salinization of freshwater are two global issues with largely serious ecological consequences. Increasing salinity in freshwater systems, as an environmental stressor, may negatively affect normal life activities in fish. It has been documented that salinity limits the invasive success of alien species by mediating physiological and life-history performances, however, there are few studies on how salinity affects its invasive process via altered behaviors. Using wild-caught invasive western mosquitofish (Gambusia affinis) as animal model, in this study, we asked whether gradual increasing salinity affects behaviors (personality and mate choice decision here), life-history traits, as well as the correlation between them by exposing G. affinis to three levels salinity (freshwater, 10 and 20‰). Results showed that, with increased salinity, male tended to be shyer, less active, less sociable, and reduced desire to mate, and female tended to be shyer, less active and lost preferences for the larger male. Furthermore, across salinity treatments, male exhibited reduced body fat content and rising reproduction allocation, however, pregnant female revealed diametrically opposed trends. In addition, the correlation between life-history traits and behaviors was only identified in pregnant female. It seems that either salinity or life-history traits directly affects mosquitofish behaviors. In summary, our results partially emphasize the harmful consequences of salinity on both life-history traits and behavioral performances. These findings provide a novel perspective on how salinity potentially affect fish fitness via altering personalities, mate choice decisions, as well as body condition, and hence supports the idea that salinity could affect the spread of invasive mosquitofish.

Out of water in the dark: Plasticity in visual structures and function in an amphibious fish.

Many fishes encounter periods of prolonged darkness within their lifetime, yet the consequences for the visual system are poorly understood. We used an amphibious fish (Kryptolebias marmoratus) that occupies dark terrestrial environments during seasonal droughts to test whether exposure to prolonged darkness diminishes visual performance owing to reduced optic tectum (OT) size and/or neurogenesis. We performed a 3-week acclimation with a 2 ×$\times $ 2 factorial design, in which fish were either acclimated to a 12 h:12 h or 0 h:24 h light:dark photoperiod in water or in air. We found that water-exposed fish had poorer visual acuity when acclimated to the dark, while air-acclimated fish had poorer visual acuity regardless of photoperiod. The ability of K. marmoratus to capture aerial prey from water followed a similar trend, suggesting that good vision is important for hunting effectively. Changes in visual acuity did not result from changes in OT size, but air-acclimated fish had 37% fewer proliferating cells in the OT than water-acclimated fish. As K. marmoratus are unable to eat on land, reducing cell proliferation in the OT may serve as a mechanism to reduce maintenance costs associated with the visual system. Overall, we suggest that prolonged darkness and air exposure can impair vision in K. marmoratus, and that changes in visual performance may be mediated, in part, by OT neurogenesis. More broadly, we show that plastic changes to the visual system of fishes can have potential consequences for organismal performance and fitness.

Context-dependent relationships between swimming, terrestrial jumping and body composition in the amphibious fish Kryptolebias marmoratus.

Understanding the mechanisms that create phenotypic variation within and among populations is a major goal of physiological ecology. Variation may be a consequence of functional trade-offs (i.e. improvement in one trait comes at the expense of another trait) or alternatively may reflect the intrinsic quality of an organism (i.e. some individuals are simply better overall performers than others). There is evidence for both ideas in the literature, suggesting that environmental context may mediate whether variation results from trade-offs or differences in individual quality. We tested this overarching 'context dependence' hypothesis by comparing the aquatic and terrestrial athletic performance of the amphibious fish Kryptolebias marmoratus captured from two contrasting habitats, a large pond and small burrows. Overall, pond fish were superior terrestrial athletes but burrow fish were better burst swimmers, suggestive of a performance trade-off at the population level. Within each population, however, there was no evidence of a performance trade-off. In burrow fish, athletic performance was positively correlated with muscle content and body condition, consistent with the individual quality hypothesis. In pond fish, there was only a relationship between glycolytic white muscle and aquatic burst performance. Notably, pond fish were in better body condition, which may mask relationships between condition and athletic performance. Overall, our data highlight that population-level trends are insufficient evidence for the existence of phenotypic trade-offs in the absence of similar within-population patterns. Furthermore, we only found evidence for the individual quality hypothesis in one population, suggesting that patterns of phenotypic covariance are context dependent.

Environmentally relevant arsenic exposure affects morphological and molecular endpoints associated with reproduction in the Western mosquitofish, Gambusia affinis.

Arsenic (As) exposure, even at low environmentally relevant levels, may cause detrimental health outcomes through developmental toxicity and by acting as an endocrine disrupting compound (EDC). Although several studies indicate that wildlife bioaccumulate As, few evaluate the health impact on fish species in their natural environment. In the U.S., As has a drinking water regulatory limit of 10 µg/L. In many parts of Arizona, surface water and groundwater have naturally elevated levels of As from geologic deposits and contamination is exacerbated by anthropogenic activity. In aquatic environments, the Western mosquitofish, Gambusia affinis, is a good bioindicator for EDC exposure because of the distinct androgen-related development of an intromittent organ, the gonapodium, in males. We evaluated morphological and reproductive outcomes in mosquitofish exposed to As. In a laboratory experiment, juvenile male mosquitofish were exposed to sodium arsenite (0 µg/L, 0.75 µg/L, 7.50 µg/L, and 75 µg/L) for 30 days, and in a field study, populations of adult male mosquitofish were collected in Arizona waterways with As levels above and below the World Health Organization's regulatory limit. In both studies, higher As exposure was significantly associated with altered hepatosomatic indices, altered fish morphology, shortened gonopodia, and lower gonopodia-somatic indices. In the field experiment, populations from surface water with higher As concentrations exhibited lower condition factors, lower gonadal-somatic indices, distinct gonopodia shapes, and altered estrogen receptor alpha and vitellogenin gene expression; androgen receptor expression was unchanged. Together, laboratory and field results suggest that As exposure at environmentally-relevant levels affects general growth and reproductive development in mosquitofish. Observed effects may further influence individual health, mobility, or reproductive function, and because G. affinis is a species known to tolerate and adapt to a wide range of environments, it serves as a local bioindicator species as well as a model organism for parallel field and laboratory studies.

Neuronal Phenotype of col4a1 and col25a1: An Intriguing Hypothesis in Vertebrates Brain Aging.

Collagens are the most abundant proteins in vertebrates and constitute the major components of the extracellular matrix. Collagens play an important and multifaceted role in the development and functioning of the nervous system and undergo structural remodeling and quantitative modifications during aging. Here, we investigated the age-dependent regulation of col4a1 and col25a1 in the brain of the short-lived vertebrate Nothobranchius furzeri, a powerful model organism for aging research due to its natural fast-aging process and further characterized typical hallmarks of brain aging in this species. We showed that col4a1 and col25a1 are relatively well conserved during vertebrate evolution, and their expression significantly increases in the brain of N. furzeri upon aging. Noteworthy, we report that both col4a1 and col25a1 are expressed in cells with a neuronal phenotype, unlike what has already been documented in mammalian brain, in which only col25a1 is considered a neuronal marker, whereas col4a1 seems to be expressed only in endothelial cells. Overall, our findings encourage further investigation on the role of col4a1 and col25a1 in the biology of the vertebrate brain as well as the onset of aging and neurodegenerative diseases.

Starter feed for carnivorous species as a practical replacement of bloodworms for a vertebrate model organism in ageing, the turquoise killifish Nothobranchius furzeri.

The absence of a controlled diet is unfortunate in a promising model organism for ageing, the turquoise killifish (Nothobranchius furzeri Jubb, 1971). Currently captive N. furzeri are fed bloodworms but it is not known whether this is an optimal diet. Replacing bloodworms with a practical dry feed would reduce diet variability. In the present study, we estimated the nutritional value of the diet ingested by wild fish and determined the fish-body amino acid profile as a proxy for their nutritional requirements. We compared the performance of fish fed four commercial feeds containing 46%-64% protein to that achieved with bloodworms and that of wild fish. Wild fish target a high-protein (60%) diet and this is supported by their superior performance on high-protein diets in captivity. In contrast, feeds for omnivores led to slower growth, lower fecundity and unnatural liver size. In comparison to wild fish, a bloodworm diet led to lower body condition, overfeeding and male liver enlargement. Out of the four dry feeds tested, the fish fed Aller matched wild fish in body condition and liver size, and was comparable to bloodworms in terms of growth and fecundity. A starter feed for carnivorous species appears to be a practical replacement for bloodworms for N. furzeri. The use of dry feeds improved performance in comparison to bloodworms and thus may contribute to reducing response variability and improving research reproducibility in N. furzeri research.