Neurotoxic effects of carbamazepine on the mosquitofish Gambusia affinis.

In recent years, the presence of pharmaceuticals in the aquatic environment has gained a significant attention. Carbamazepine, a commonly prescribed antiepileptic drug, has been consistently found in aquatic environments at concentrations ranging from nanograms to micrograms, raising concerns about its potential negative impacts on aquatic organisms. The study examined the acute and chronic neurotoxic effects of environmentally relevant and sublethal concentrations of carbamazepine in the mosquitofish Gambusia affinis. After a 96-hour exposure period, the median lethal concentration (LC50) of carbamazepine for G. affinis was determined as 24 mg L - 1. For the current study, sublethal concentrations i.e., one-tenth (2.4 mg L - 1) and one-fifth (4.8 mg L - 1) of the LC50 value were chosen for assessing the neurotoxic effects along with the environmentally relevant concentration (13 ng L - 1). The research findings indicated that carbamazepine had a disruptive impact on the typical growth and behavior of the fish. During the acute exposure phase, physical deformities were observed in the fish, resulting in neonatal and postneonatal fatalities. Furthermore, the neurotoxic effects of carbamazepine were clearly demonstrated through alterations in various neurological parameters, including acetylcholinesterase, dopamine, gamma-aminobutyric acid, serotonin, monoamine oxidase, 5-hydroxyindole acetic acid, adrenaline, and nor-adrenaline. These findings raise concerns about the survival of fish populations in their natural environment.

Battle of the Bites: The Effect of Sewage Effluent Exposure on Mosquitofish Biocontrol of Mosquitoes in Residential Louisiana.

Mosquitofish, Gambusia affinis, are eponymous larval mosquito predators. Their ability to colonize and survive in habitats that are uninhabitable by other potential predators allows them to naturally manage larval mosquito populations in most ground pools they are present in. However, effluent from residential onsite wastewater treatment systems (OWTSs) appears to limit the presence of fish predators. This is especially problematic in Louisiana, where regulations allow the discharge of OWTS effluent into open drainage conveyances. To determine the effect of effluent on the capacity of mosquitofish for biocontrol in contaminated areas, we assessed the body condition metrics of populations from two effluent-exposed sites and two sites not exposed to effluent, determined the lethal effect of effluent-contaminated drainage water on fish, and measured the prey consumption rates in the presence of effluent. Female fish collected from effluent-impacted sites had a reduced somatic body condition and most females examined displayed masculinized anal fins resembling the male gonopodium structure. This trait was not seen in fish collected from the control sites and has not yet been documented in association with OWTSs or in the state of Louisiana. Fish from the control sites survived at effluent-contaminated water levels < 70%, and the prey clearance rates increased with dilution. Onsite wastewater treatment system effluent has significant effects on both the short- and long-term persistence of mosquitofish, their body composition, reproductive health, and larval mosquito consumption. These effects likely release mosquito larvae from suppression and may increase the threat of mosquito-transmitted pathogens in effluent-contaminated locations.

Species distribution modeling combined with environmental DNA analysis to explore distribution of invasive alien mosquitofish (Gambusia affinis) in China.

China has become one of the most serious countries suffering from biological invasions in the world. In the context of global climate change, invasive alien species (IAS) are likely to invade a wider area, posing greater ecological and economic threats in China. Western mosquitofish (Gambusia affinis), which is known as one of the 100 most invasive alien species, has distributed widely in southern China and is gradually spreading to the north, causing serious ecological damage and economic losses. However, its distribution in China is still unclear. Hence, there is an urgent need for a more convenient way to detect and monitor the distribution of G. affinis to put forward specific management. Therefore, we detected the distribution of G. affinis in China under current and future climate change by combing Maxent modeling prediction and eDNA verification, which is a more time-saving and reliable method to estimate the distribution of species. The Maxent modeling showed that G. affinis has a broad habitat suitability in China (especially in southern China) and would continue to spread in the future with ongoing climate change. However, eDNA monitoring showed that occurrences can already be detected in regions that Maxent still categorized as unsuitable. Besides temperature, precipitation and human influence were the most important environmental factors affecting the distribution of G. affinis in China. In addition, by environmental DNA analysis, we verified the presence of G. affinis predicted by Maxent in the Qinling Mountains where the presence of G. affinis had not been previously recorded.

Non-concordant epigenetic and transcriptional responses to acute thermal stress in western mosquitofish (Gambusia affinis).

Climate change is intensifying the frequency and severity of extreme temperatures. Understanding the molecular mechanisms underlying the ability to cope with acute thermal stress is key for predicting species' responses to extreme temperature events. While many studies have focused on the individual roles of gene expression, post-transcriptional processes and epigenetic modifications in response to acute thermal stress, the relative contribution of these molecular mechanisms remains unclear. The wide range of thermal limits of western mosquitofish (Gambusia affinis) provides an opportunity to explore this interplay. Here, we quantified changes in gene expression, alternative splicing, DNA methylation and microRNA (miRNA) expression in muscle tissue dissected from mosquitofish immediately after reaching high (CTmax) or low thermal limit (CTmin). Although the numbers of genes showing expression and splicing changes in response to acute temperature stress were small, we found a possibly larger and non-redundant role of splicing compared to gene expression, with more genes being differentially spliced (DSGs) than differentially expressed (DEGs), and little overlap between DSGs and DEGs. We also identified a small proportion of CpGs showing significant methylation change (i.e. differentially methylated cytosines, DMCs) in fish at thermal limits; however, there was no overlap between DEGs and genes annotated with DMCs in both CTmax and CTmin experiments. The weak interplay between epigenetic modifications and gene expression was further supported by our discoveries of no differentially expressed miRNAs. These findings provide novel insights into the relative role of different molecular mechanisms underlying immediate responses to extreme temperatures and demonstrate non-concordant responses of epigenetic and transcriptional mechanisms to acute temperature stress.

Gestodene causes masculinization of the western mosquitofish (Gambusia affinis): Insights from ovary metabolomics.

Gestodene (GES) is a common synthetic progesterone frequently detected in aquatic environments. Chronic exposure to GES can cause masculinization of a variety of fish; however, whether metabolism is closely related to the masculinization has yet to be explored. Hence, the ovary metabolome of adult female western mosquitofish (Gambusia affinis) after exposing to GES (0.0, 5.0, 50.0, and 500.0 ng/L) for 40 days was analyzed by using high-performance liquid chromatography ionization with quadrupole time-of-flight tandem mass spectrometry (HPLC-QTOF-MS). The results showed that GES increased the levels of cysteine, taurine, ophthalmic acid and cAMP while decreased methionine, these metabolites changes may owing to the oxidative stress of the ovaries; while taurcholic acid and uric acid were decreased along with induced oocyte apopotosis. Steroids hormone metabolism was also significantly affected, with progesterone and cortisol being the most affected. Enzyme-linked immunoassay results showed that estradiol levels were decreased while testosterone levels were increased with GES exposure. In addition, correlation analysis showed that the differential metabolites of some amino acids (e.g. leucine) were strongly correlated with the levels of steroids hormones secreted by the pituitary gland. The results of this study suggest that GES affects ovarian metabolism via the hypothalamus-pituitary-gonad and hypothalamic-pituitary-adrenal axes, impair antioxidant capacity, induce apoptosis in the ovary of G. affinis, and finally caused masculinization.

Effects of norethindrone on the growth, behavior, and thyroid endocrine system of adult female western mosquitofish (Gambusia affinis).

Progestins are mainly used in pharmacotherapy and animal husbandry and have received increasing attention as they are widely detected in various aquatic ecosystems. In this study, adult female western mosquitofish (Gambusia affinis) were exposed to different concentrations of norethindrone (NET) (solvent control, 5.0 (L), 50.0 (M), and 500.0 (H) ng/L) for 42 days. Behaviors, morphological parameters, histology of the thyroid, thyroid hormone levels (TSH, T3, and T4), and transcriptional levels of nine genes in the hypothalamic-pituitary-thyroid (HPT) axis were examined. The results showed that NET decreased sociality but increased the anxiety of G. affinis. Sociality makes fish tend to cluster, and anxiety may cause G. affinis to reduce exploration of new environments. Female fish showed hyperplasia, hypertrophy, and glial depletion in their thyroid follicular epithelial cells after NET treatment. The plasma levels of TSH and T4 were significantly reduced, but T3 concentrations were significantly increased in the fish from the H group. In addition, the transcripts of genes (tshb, tshr, tg, dio1, dio2, thrb) in the brains of fish in the M and H treatments were significantly stimulated, while those of trh and pax2a were suppressed. Our results suggest that NET may impact key social behaviors in G. affinis and interfere with the entire thyroid endocrine system, probably via affecting the transcriptional expression of upstream regulators in the HPT axis.

Transfer of Se from sediments to the western mosquitofish Gambusia affinis: Tissue distribution, accumulation, and effects on the antioxidant physiology.

Selenium (Se) has been shown to cause various toxicities in predatory species (i.e., fish and birds) in Se-contaminated aquatic environments. However, trophic transfer of Se from abiotic environments to freshwater fish has been relatively less addressed. In this study, 2-month-old mosquitofish (Gambusia affinis) were fed Se-enriched oligochaete (Lumbriculus variegatus, exposed to different concentrations of Se(IV) at 0.0, 3.0, 10.0, and 30.0 µg/g dry weight for 7 days) for 45 days. Tissue distribution, Se speciation, and effects on the antioxidant physiology in G. affinis were assessed. The results showed Se was rapidly accumulated in the oligochaete, with 6.30 ± 1.20, 16.20 ± 2.10, and 34.50 ± 2.40 µg/g dw of total Se levels in the worms exposed to 3.0, 10.0, and 30.0 µg/g of Se(IV), respectively. Total Se levels were increased in a dose-dependent manner in fish tissues and Se(IV) from sediments was maternally transferred to the fish embryos. Se-Met-and Se-Cys-were the predominant Se species in the worm and fish tissues, accounting for a minimum of 91.01% of the total Se. Furthermore, increased lipid peroxidation and altered the activities of antioxidant enzymes and levels of GSH were noticed in G. affinis fed the Se-enriched L. variegatus. This study has demonstrated that Se(IV) is transferred from an abiotic vector to freshwater organisms, disturbing the antioxidant physiology in G. affinis and potentially their offspring. This study highlights the importance of dietary exposure on the accumulation and toxicity of Se in aquatic organisms.

Effects of TLR2/4 signalling pathway in western mosquitofish (Gambusia affinis) after Edwardsiella tarda infection.

Gambusia affinis is regarded as an important animal model. Edwardsiella tarda is one of the most serious pathogens affecting aquaculture. The study explores the effects of TLR2/4 partial signalling pathway in G. affinis of E. tarda infection. The study collected the brain, liver, and intestine after E. tarda LD50 and 0.85% NaCl solution challenge at different times (0 h, 3 h, 9 h, 18 h, 24 h, and 48 h). In these three tissues, the mRNA levels of PI3K, AKT3, IRAK4, TAK1, IKKß, and IL-1ß were substantially enhanced (p < .05) then returned to normal levels. Additionally, Rac1 and MyD88 in liver had different trend with other genes in brain and intestine, which displayed significantly indifference. The overexpression of IKKß, and IL-1ß indicated that E. tarda also caused immune reaction in intestine and liver, which would be consistent with delayed edwardsiellosis, which causes intestinal lesions and liver and kidney necrosis. Additionally, MyD88 plays a smaller role than IRAK4 and TAK1 in this signalling pathways. This study could enrich the understanding of the immune mechanism of the TLR2/4 signalling pathway in fish and might help to prescribe preventive measures against E. tarda to prevent infectious diseases in fish.

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.

Comprehensive evaluation of the toxicity of the flame retardant (decabromodiphenyl ether) in a bioindicator fish (Gambusia affinis).

In recent years, concerns have increased about the adverse effects on health and the environment of polybrominated diphenyl ethers (PBDEs), especially BDE-209, the most widely PBDE used globally. These pollutants derive from e-waste and present different adverse effects on biota. In this work, a toxicological study on mosquitofish (Gambusia affinis) using BDE-209 (2,2',3,3',4,4',5,'5',6,6'-decabromodiphenyl ether) was carried out. Acute toxicity bioassays were conducted with daily renewal of solutions, using different concentrations of environmental relevance, ranged between 10 and 100 µg L-1 of BDE-209. At 48 and 96 h of exposure, several parameters were evaluated, such as mortality, individual activity (swimming), biochemical activity (catalase; thiobarbituric acid-reactive substances; and acetylcholinesterase), and cytotoxic responses (micronucleus frequencies). In addition, integrated biomarker response and multivariate analyses were conducted to study the correlation of biomarkers. The calculated Lethal Concentration-50 remained constant after all exposure times (24 to 96 h), being the corresponding value 27.79 µg L-1 BDE-209. Furthermore, BDE-209 induced effects on the swimming activity of this species in relation to acetylcholine, since BDE-209 increased, producing oxidative damage at the biochemical level and genotoxicity after 48 h of exposure to 10 and 25 µg L-1 BDE-209. The results indicate that BDE-209 has biochemical, cytotoxic, neurotoxic, and genotoxic potential on G. affinis. In addition, mosquitofish could be used as a good laboratory model to evaluate environmental stressors since they could represent a risk factor for Neotropical species.

Color Discrimination Provides Insight into the Relationship between Personality Cognition and Brain Morphology in the Western Mosquitofish (Gambusia affinis).

Recent studies on the differences in cognitive ability between individuals focused on two aspects: one is whether the individual differences in cognitive ability are related to brain size, the other is whether they pertain to certain personality traits. To explore these two hypotheses, we tested the personality traits, cognitive abilities, and brain volumes of western mosquitofish (Gambusia affinis). First, a color preference test was conducted to select two unbiased colors for G. affinis for subsequent cognitive tests. The results showed that G. affinis had a great preference for red and green to yellow and blue; therefore, the red-green combination was selected for the study of cognitive abilities. Then, we explored the relationship among cognition, personality, and brain morphology through cognitive abilities tests, personality traits, and brain volume measurements. We found that there was a trade-off among cognition, personality, and brain morphology. For example, more active individuals found food faster, but had also poor memory; Those individuals with larger corpus cerebelli were bolder while they were less likely to find food; The individuals that found food faster were more active and had a smaller inferior lobe. The color preference test provides a reliable way for selecting unbiased colors for behavioral studies in G. affinis. Meanwhile, our study indicates that there exists a balance mechanism among cognition, personality, and brain morphology.

Drivers and Cascading Ecological Consequences of Gambusia affinis Trait Variation.

AbstractPhenotypic trait differences among populations can shape ecological outcomes for communities and ecosystems. However, few studies have mechanistically linked heritable and plastic components of trait variation to generalizable processes of ecology, such as trophic cascades. Here, we assess morphological and behavioral trait variation in nine populations of common garden-reared western mosquitofish (Gambusia affinis) from three distinct ancestral predator environments (three populations per environment), each reared in the presence and absence of predator cues. We then use a pond mesocosm experiment to examine the ecological consequences of mosquitofish trait variation and density variation. Our results show significant among-population trait variation, but this variation was generally unrelated to ancestral predator environment. When traits did vary congruently with respect to ancestral predator environment, this trait variation was driven by gene-by-environment interactions. Variation in several mosquitofish traits altered the cascading effects of mosquitofish on zooplankton and primary producers, but the effect of any given trait was typically weaker than that of density. We note that the relatively stronger ecological effects of density may mask the effects of traits in some systems. Our example here shows that trait variation can be highly noncongruent with respect to a perceived selective agent, phenotypic change is a product of complex interactions between genes and the environment, and numerous interacting phenotypes generate significant but potentially cryptic cascading ecological change.

The legacy of predator threat shapes prey foraging behaviour.

Predators exert strong selection on prey foraging behaviour such that prey responses may reflect a combination of ancestral effects of predators (genetic and nongenetic transgenerational effects), past individual experience with predators (phenotypic plasticity), and current exposure to predators (behavioural response). However, the importance of these factors in shaping prey foraging behaviour is not well understood. To test the relative effects of ancestry, prior experience, and current exposure, we measured foraging rates and food size preference of different ancestry and exposure groups of Western mosquitofish in the presence and absence of immediate threat from predatory largemouth bass. Our results confirm that mosquitofish had lower foraging rate in the immediate presence of predator threat. Mosquitofish also foraged at a lower rate if they had ancestry with predators, regardless of immediate threat. In contrast, individual prior experience with predators only caused reduced foraging rates in the immediate presence of a predator. This suggests that phenotypic plasticity could carry a lower risk of maladaptive antipredator responses-i.e., reduced food intake-in the complete absence of a predator. Finally, in the presence of a predator, mosquitofish with both ancestry and experience with predators consumed larger, presumably more energetically valuable, food items. Overall, our results show that non-consumptive effects of predators on prey behaviour can persist within and across generations, such that the legacy of past predator exposure-or "the ghost of predation past"-may continue to shape prey behaviour even when predators are no longer around.

Invasive fish retain plasticity of naturally selected, but diverge in sexually selected traits.

Invasive alien species (IAS) have become a major threat to ecosystems worldwide. From an evolutionary ecological perspective, they allow teasing apart the relative contributions of plasticity and evolutionary divergence in driving rapid phenotypic diversification. When IAS spread across extensive geographic ranges, climatic variation may represent a source of strong natural selection through overwinter mortality and summer heat stress. This could favour local adaptation, i.e., evolutionary divergence of certain traits. IAS, however, are likely to show plasticity in survival-related traits, and environmental fluctuation in their new distribution range could favour the maintenance of this pre-existing phenotypic plasticity. By contrast, sexually selected traits are more likely to undergo evolutionary divergence when components of sexual selection differ geographically. Here, using data from a common-garden rearing experiment of Western mosquitofish (Gambusia affinis Baird and Girard, 1853) from five populations across the species' invasive range in China, we show that invasive mosquitofish have retained plasticity in key physiological (thermal tolerances), morphological and life-history traits even 100 years after their introduction to China, but exhibit heritable population differences in several sexually selected traits, including the shape of the male copulatory organ. Adaptive plasticity of traits linked to immediate survival in different thermal environments-while likely responsible for the species' extraordinary invasion success-could slow down genetic evolution. Several sexually selected traits could diverge geographically and show rapid evolutionary change, e.g., because climate alters selective landscapes arising from mate competition as an indirect consequence of variation in overwinter mortality.

Consumer trait responses track change in resource supply along replicated thermal gradients.

Rising temperatures may alter consumer diets through increased metabolic demand and altered resource availability. However, current theories assessing dietary shifts with warming do not account for a change in resource availability. It is unknown whether consumers will increase consumption rates or consume different resources to meet increased energy requirements and whether the dietary change will lead to associated variation in morphology and nutrient utilization. Here, we used populations of Gambusia affinis across parallel thermal gradients in New Zealand (NZ) and California (CA) to understand the influence of temperature on diets, morphology and stoichiometric phenotypes. Our results show that with increasing temperature in NZ, mosquitofish consumed more plant material, whereas in CA mosquitofish shifted towards increased consumption of invertebrate prey. In both regions, populations with plant-based diets had fuller guts, longer relative gut lengths, better-orientated mouths and reduced body elemental %C and N/P. Together, our results show multiple pathways by which consumers may alter their feeding patterns with rising temperatures, and they suggest that warming-induced changes to resource availability may be the principal determinant of which pathway is taken.

Serotonin (5-hydroxytryptamine)-immunoreactive neurons in the brain of the viviparous fish Gambusia affinis.

The monoaminergic neurotransmitter serotonin (5-HT) acts as a neuromodulator and is associated with a wide range of functions in fish. In this investigation, 5-HT immunoreactivity was studied in the central nervous system (CNS) of the viviparous mosquitofish Gambusia affinis. 5-HT-immunoreactive (5-HT-ir) cells/fibres were observed throughout the subdivisions of ventral and dorsal telencephalon including the olfactory bulb. Several intensely stained 5-HT-ir cells and/or fibres were detected in different areas of the hypothalamus as well as the proximal pars distalis of the pituitary gland. 5-HT-ir cells were restricted to the dorsal and ventral part of the pretectal diencephalic cluster, but only fibres were detected in the anterior, ventromedial and posterior subdivisions of the thalamic nucleus and in the preglomerular complex. In the mesencephalon, 5-HT-ir perikarya, and fibres were seen in the optic tectum, midbrain tegmentum and torus semicircularis. A cluster of prominently labelled 5-HT-ir neurons was observed in the superior raphe nucleus, whereas numerous 5-HT-ir fibres were distributed throughout the rhombencephalic divisions. In addition, a bundle of rostrocaudally running 5-HT-ir fibres was noticed in the spinal cord. This is the first detailed neuroanatomical study in a viviparous teleost, reporting a widespread distribution of 5-HT-ir somata and fibres in the CNS. The results of this study provide new insights into the evolutionarily well conserved nature of the monoaminergic system in the CNS of vertebrates and suggest a role for 5-HT in regulation of several physiological, behavioural and neuroendocrine functions in viviparous teleosts.

Male size and reproductive performance in three species of livebearing fishes (Gambusia spp.): A systematic review and meta-analysis.

The genus Gambusia represents approximately 45 species of polyandrous livebearing fishes with reversed sexual size dimorphism (i.e. males smaller than females) and with copulation predominantly via male coercion. Male body size has been suggested as an important sexually selected trait, but despite abundant research, evidence for sexual selection on male body size in this genus is mixed. Studies have found that large males have an advantage in both male-male competition and female choice, but that small males perform sneaky copulations better and at higher frequency and thus may sire more offspring in this coercive mating system. Here, we synthesized this inconsistent body of evidence using pre-registered methods and hypotheses. We performed a systematic review and meta-analysis of summary and primary (raw) data combining both published (n = 19 studies, k = 106 effect sizes) and unpublished effect sizes (n = 17, k = 242) to test whether there is overall selection on male body size across studies in Gambusia. We also tested several specific hypotheses to understand the sources of heterogeneity across effects. Meta-analysis revealed an overall positive correlation between male size and reproductive performance (r = 0.23, 95% confidence interval: 0.10-0.35, n = 36, k = 348, 4,514 males, three Gambusia species). Despite high heterogeneity, the large-male advantage appeared robust across all measures studied (i.e. female choice, mating success, paternity, sperm quantity and quality), and was considerably larger for female choice (r = 0.43, 95% confidence interval: 0.28-0.59, n = 14, k = 43). Meta-regressions found several important factors explaining heterogeneity across effects, including type of sperm characteristic, male-to-female ratio, female reproductive status and environmental conditions. We found evidence of publication bias; however, its influence on our estimates was attenuated by including a substantial amount of unpublished effects, highlighting the importance of open primary data for more accurate meta-analytic estimates. In addition to positive selection on male size, our study suggests that we need to rethink the role and form of sexual selection in Gambusia and, more broadly, to consider the ecological factors that affect reproductive behaviour in livebearing fishes.

The pivotal protein profile between the conjoined twins and normal mosquitofish Gambusia affinis based on iTRAQ proteomic analysis.

The fish abnormal embryonic development has attracted public attention in the recent few years. In this study, an iTRAQ proteomic analysis of mosquitofish between conjoined twins and normal fishes is applied for the first time by using the genome database of mosquitofish. Three thousand four hundred ninety proteins were identified with 304 differentially expressed proteins (DEPs). One hundred six differentially upregulated proteins (DUPs) and 198 differentially downregulated proteins (DDPs) were identified between the conjoined twins and normal mosquitofish groups. Notably, the proteins related to lipid and proteolysis were the important GO terms for the DUPs while response to light stimulus and response to radiation were the most enriched GO terms for the DDPs. The proteins related to lysosome, apoptosis, autophagy, and phagosome were the functional KEGG pathway for the DUPs while most of the pathways were related to cardiovascular for the DDPs. This study expatiated a pivotal protein profile between the conjoined twins and normal mosquitofish which can provide a conference for fish embryonic development.

Synonymous single nucleotide polymorphism in arsenic (+3) methyltransferase of the Western mosquitofish (Gambusia affinis) and its gene expression among field populations.

Naturally occurring arsenic is toxic at extremely low concentrations, yet some species persist even in high arsenic environments. We wanted to test if these species show evidence of evolution associated with arsenic exposure. To do this, we compared allelic variation across 872 coding nucleotides of arsenic (+3) methyltransferase (as3mt) and whole fish as3mt gene expression from three field populations of Gambusia affinis, from water sources containing low (1.9 ppb), medium-low (3.3 ppb), and high (15.7 ppb) levels of arsenic. The high arsenic site exceeds the US EPA's Maximum Contamination Level for drinking water. Medium-low and high populations exhibited homozygosity, and no sequence variation across all animals sampled. Eleven of 24 fish examined (45.8%) in the low arsenic population harbored synonymous single nucleotide polymorphisms (SNPs) in exons 4 and/or 10. SNP presence in the low arsenic population was not associated with differences in as3mt transcript levels compared to fish from the medium-low site, where SNPs were noted; however, as3mt expression in fish from the high arsenic concentration site was significantly lower than the other two sites. Low sequence variation in fish populations from sites with medium-low and high arsenic concentrations suggests greater selective pressure on this allele, while higher variation in the low population suggests a relaxed selection. Our results suggest gene regulation associated with arsenic detoxification may play a more crucial role in influencing responses to arsenic than polymorphic gene sequence. Understanding microevolutionary processes to various contaminants require the evaluation of multiple populations across a wide range of pollution exposures.

Effects of dexamethasone on the morphology, gene expression and hepatic histology in adult female mosquitofish (Gambusia affinis).

Glucocorticoids (GCs), including natural hormones as well as synthetic chemicals, can pose influences on physiological performance, development and reproduction of fish. Dexamethasone (DEX) is a synthetic glucocorticoid widely used as pharmaceutical and usually exists in effluents with varying degrees of concentrations. In this study, adult female mosquitofish (Gambusia affinis) were treated by DEX at concentrations of 0, 0.5, 5 and 50 µg/L for 60 days. Morphological parameters of anal fin and skeleton, mRNA expression abundance, and histological alterations of liver were investigated to assess effects of DEX on mosquitofish. The results showed that DEX increased number of sections of ray 3 in anal fin and decreased 16L, 15D and 16D in skeletal parameters, which indicates DEX could potentially lead to weak masculinization. Furthermore, transcriptional expression levels of ARα, ARß, ERß, VTGC and CYP19A genes were notably down-regulated by DEX, which will contribute to weak masculinization in females. In addition, the damage to liver tissue was also induced by DEX. Taken together, this research demonstrated that aquatic environments contaminated by DEX have negative effects on mosquitofish at a population level.