Vitellogenin Levels Research Articles

Polystyrene microplastics increase estrogenic effects of 17α-ethynylestradiol on male marine medaka (Oryzias melastigma)

Microplastics (MPs) and endocrine disrupting chemicals are ubiquitous pollutants in marine environments, but their combined ecological risk is unclear. This study exposed male marine medaka (Oryzias melastigma) to 10 ng/L 17α-ethynylestradiol (EE2) alone or EE2 plus 2, 20, and 200 μg/L polystyrene MPs for 28 days to investigate the impacts of MPs on the reproductive disruption of EE2. The results showed that 10 ng/L EE2 alone did not affect biometric parameters, while co-exposure to EE2 and 20, 200 μg/L MPs suppressed the growth and decreased gonadosomatic and hepatosomatic indices. Compared to EE2 alone, EE2 plus MPs exposure significantly increased plasma 17β-estradiol (E2) levels in a dose-dependent manner, and co-exposure to EE2 and 20, 200 μg/L MPs significantly increased the ratios of E2/testosterone (T). Moreover, EE2 plus MPs exposure elevated the transcription levels of estrogen biomarker genes vitellogenin and choriogenin, and estrogen receptor (ERα and ERβ). Morphological analysis also showed that co-exposure to EE2 and MPs induced more severe damage to the testes and livers, indicating that MPs increased the toxicity of EE2. The actual EE2 concentrations in the solution increased with the exposure concentrations of MPs, suggesting that MPs changed the fate and behavior of EE2 in the seawater. These findings demonstrate that MPs could increase the estrogenic effects of EE2 on marine fish, suggesting that the combined health risk of MPs and endocrine disrupting chemicals on marine organisms should be paid great attention.

Impact of bisphenol-A and synthetic estradiol on brain, behavior, gonads and sex hormones in a sexually labile coral reef fish

Endocrine disrupting chemicals, such as bisphenol A (BPA) and ethinylestradiol (EE2), are detected in the marine environment from plastic waste and wastewater effluent. However, their impact on reproduction in sexually labile coral reef fish is unknown. The objective of this study was to determine impacts of environmentally relevant concentrations of BPA and EE2 on behavior, brain gene expression, gonadal histology, sex hormone profile, and plasma vitellogenin (Vtg) levels in the anemonefish, Amphiprion ocellaris. A. ocellaris display post-maturational sex change from male to female in nature. Sexually immature, male fish were paired together and fed twice daily with normal food (control), food containing BPA (100 μg/kg), or EE2 (0.02 μg/kg) (n = 9 pairs/group). Aggression toward an intruder male was measured at 1, 3, and 6 months. Blood was collected at 3 and 6 months to measure estradiol (E2), 11-ketotestosterone (11-KT), and Vtg. At the end of the study, fish were euthanized to assess gonad morphology and to measure expression of known sexually dimorphic genes in the brain. Relative to control, BPA decreased aggression, altered brain transcript levels, increased non-vitellogenic and vitellogenic eggs in the gonad, reduced 11-KT, and increased plasma Vtg. In two BPA-treated pairs, both individuals had vitellogenic eggs, which does not naturally occur. EE2 reduced 11-KT in subordinate individuals and altered expression of one transcript in the brain toward the female profile. Results suggest BPA, and to a lesser extent EE2, pollution in coral reef ecosystems could interfere with normal reproductive physiology and behavior of the iconic sexually labile anemonefish.

Expression profiles of hepatic vitellogenin and gonadal zona pellucida subtypes in gray mullet (Mugil cephalus) with 17α-ethinylestradiol-induced gonadal abnormality.

The subtypes of zona pellucida (zp), primarily expressed in female gonads, are considered novel molecular markers for testis-ova (or intersex), a type of gonadal abnormality caused by environmental estrogens (EEs) in Japanese medaka (Oryzias latipes). However, the association between testis-ova and the expression of gonadal zp subtypes is unclear in other teleost species, particularly in species studied in field surveys. In this study, 17α-ethinylestradiol (EE2) was orally administrated at 4-4000ng/g body weight (BW)/day for 28 days to gray mullets (Mugil cephalus), and gonadal abnormalities were studied using histological analysis. The expression profiles of gonadal zp subtypes (zpb and zpc5) were analyzed to evaluate their suitability as gonadal abnormality markers by comparing with a hepatic vitellogenin (vtg) subtype (vtgAb). The oral administration of EE2 40 and 400ng/g BW/day for 28 days induced significant gonadal zpb expression, and the gonads showed moderate abnormality (testis-ova). Conversely, the gonadal zpc5 levels decreased significantly in response to the oral administration of EE2 at 4000ng/g BW/day for 28 days, and the gonads exhibited severe abnormalities. The hepatic vtgAb levels increased upon EE2 treatment regardless of gonadal abnormality. Therefore, the gonadal zpb levels and hepatic vtgAb levels served as appropriate markers for testis-ova and EE2 presence, respectively. However, the diagnosis of severe gonadal abnormality using gonadal zpc5 was moderately accurate. The findings suggest that the combination of vtgAb, zpb, and zpc5 is a potential marker for gonadal abnormality caused by EE contamination in gray mullet. That said, the potential of zpc5 should be reconsidered to determine if it shows greater accuracy in a larger or more diverse population.

Biodegradable polymers and their nano-composites for the removal of endocrine-disrupting chemicals (EDCs) from wastewater: A review

Endocrine-disrupting chemicals (EDCs) target the endocrine system by interfering with the natural hormones in the body leading to adverse effects on human and animal health. These chemicals have been identified as major polluting agents in wastewater effluents. Pharmaceuticals, personal care products, industrial compounds, pesticides, dyes, and heavy metals are examples of substances that could be considered endocrine active chemicals. In humans, these chemicals could cause obesity, cancer, Alzheimer's disease, autism, reproductive abnormalities, and thyroid problems. While in wildlife, dysfunctional gene expression could lead to the feminization of some aquatic organisms, metabolic diseases, cardiovascular risk, and problems in the reproductive system as well as its levels of hatchability and vitellogenin. EDCs could be effectively removed from wastewater using advanced technologies such as reverse osmosis, membrane treatment, ozonation, advanced oxidation, filtration, and biodegradation. However, adsorption has been proposed as a more promising and sustainable method for water treatment than any other reported technique. Increased attention has been paid to biodegradable polymers and their nano-composites as promising adsorbents for the removal of EDCs from wastewater. These polymers could be either natural, synthetic, or a combination of both. This review presents a summary of the most relevant cases where natural and synthetic biodegradable polymers have been used for the successful removal of EDCs from wastewater. It demonstrates the effectiveness of these polymers as favorable adsorbents for novel wastewater treatment technologies. Hitherto, very limited work has been published on the use of both natural and synthetic biodegradable polymers to remove EDCs from wastewater, as most of the studies focused on the utilization of only one type, either natural or synthetic. Therefore, this review could pave the way for future exploration of biodegradable polymers as promising and sustainable adsorbents for the removal of various types of pollutants from wastewater.

Chlorpyrifos Effect on Vitellogenin, Ovarian Steroid in Adult and NR5A1 Expression in Fry of the Freshwater Catfish, Heteropneustes fossilis (Bloch, 1794)

Aim: To study the interference of organophosphate pesticide chlorpyrifos (CPF) in gonad function at the steroidal and gene levels during prespawning adult and fry stage respectively of the freshwater stinging catfish, Heteropneustes fossilis. Methods: The matured adult fish of early prespawning stage were exposed to different combinations of CPF (0.174 μM, 10-1th of 96 h LC50), gonadotropin (hCG; 100 IU.fish-1) and natural antioxidants (curcumin; 10 mg.L-1 and ascorbic acid; 5 mg.L-1) for 24 h to measure vitellogenin and ovarian steroid. Twenty days post-hatch, fish fry were exposed to CPF for 96 h to understand the influence on NR5A1 gene expression pattern. Results: The CPF increased vitellogenin level at liver, ovary and serum. Among steroidal cascade CPF induced highest estradiol followed by a significant increase in progesterone and 17α, 20β-dihydroxyprogesterone as compared to control fish. The gonadotropin further increased CPF induced effects on vitellogenin and ovarian steroidal level. Though, antioxidants co-incubation with CPF did not produce any significant change on either vitellogenin or in ovarian steroids content. CPF triggers NR5A1 gene expression in a fry that indicates a role in early gonad differentiation and development. Conclusion: The present study clearly indicates the major influence of CPF in gonadal physiology of both adult and fry stages of freshwater catfish, Heteropneustes fossilis.

Study of Heavy Metals Pollution and Vitellogenin Levels in Brown Trout (Salmo trutta trutta) Wild Fish Populations

The objectives of this research were, first, to determine the concentrations of certain heavy metals in the edible tissue of wild brown trout (Salmo trutta trutta) from two different rivers located in the Abruzzi region (Italy), and then, to investigate the levels of variation in vitellogenin (VTG) associated with the presence of metalloestrogens. VTG is an effective indicator for endocrine disturbance, and an increase in the vitellogenin levels in male fish is widely employed as a biomarker of estrogenic contamination in the aquatic environment. The muscles of the trout were analyzed for As, Cd, Co, Cr, Ni, Pb, Al, and Zn using an inductively coupled plasma-mass spectrometer (ICP-MS), and Hg was measured using a direct mercury analyzer (DMA-80). The calculated values of the condition factor confirmed a healthy status for this species, indicating that the aquatic habit in both rivers is suitable for brown trout life. No significant difference in the concentrations of each metal were reported between the trout from the two rivers, and no significant difference for VTG levels were found between male and female fish. It is interesting to note the high concentrations of Al/Zn, while the Pb, Cd, and Hg concentrations in all of the samples were lower than those established by the European Commission.

Effects of continuous and intermittent cadmium exposure on HPGL axis, GH/IGF axis and circadian rhythm signaling and their consequences on reproduction in female zebrafish: Biomarkers independent of exposure regimes

Typical biomarkers of cadmium (Cd) pollution have well been confirmed in fish from continuous exposure pattern. However, in a natural environment, fish may be exposed to Cd intermittently. In this study, juvenile female zebrafish were exposed for 48 days to 10 μg/L Cd continuously, 20 μg/L for 1 day in every 2 days or 30 μg/L for 1 day in every 3 days. The toxic effects were evaluated using 8 various physiological and biochemical endpoints like specific growth rate (SGR), 17β-estradiol (E2) and vitellogenin (VTG) concentrations in plasma, reproductive parameters (gonadosomatic index (GSI), egg-laying amount, spawning percentage, and hatching and mortality rate of embryos). Transcription of 59 genes related to hypothalamic-pituitary-gonadal-liver (HPGL) axis, circadian rhythm signaling and insulin-like growth factor (IGF) system was examined. SGR, spawning percentage, E2 and VTG levels declined in fish exposed to 10 and 20 μg/L Cd but remained relatively stable in fish exposed to 30 μg/L Cd. Exposure to 10, 20 and 30 μg/L Cd significantly reduced GSI, hatching rate and mortality rate. Similarly, mRNA expression of 27 genes were sensitive to both continuous and intermittent Cd exposure. Among these genes, expression levels of 10 genes had more than 5-fold increase or decrease, including mRNA levels of vtg1, vtg2, vtg3, esr1, igf2a, igf2b, igfbp5b, nr1d1, gnrh3 and gnrhr4. The most sensitive molecular biomarker was vtg3 expression with 1500–3100 fold increase in the liver. The present study, for the first time, provides effective candidate biomarkers for Cd, which are independent of exposure regimes.

Morpho-meristics, maturity stages, GSI and gonadal hormone plasticity of African catfish Clarias gariepinus (Burchell 1822) that invaded into the Ganga River, India

BackgroundAfrican catfish Clarias gariepinus introduced to India has gravitated into the Ganga River as an invasive species. Morphological plasticity and reproductive adaptation are considered and reported as important manifestation contributing to evolution and persistence of an invasive species in the novel environment facilitating its expansion and establishment. African catfish in the Ganga River although documented to exist, it is yet to investigate if it elicits adaptation responses through morphological and reproductive plasticity in the riverine conditions. Therefore, morpho-meristic changes, plasticity in the reproductive stages, gonadosomatic index (GSI) and gonadal hormones were examined in C. gariepinus that invaded into the Ganga River so as to ascertain its invasion success.ResultsOut of 23 morpho-meristic characters examined, head length (HL), head depth (HD), anal fin length (AFL) and the pectoral fin rays (PECFR) were observed to differ significantly (p<0.05). The correlation coefficient ‘r’ between log length and log weight was found to be 0.9690 in culture and 0.8754 in river-caught specimens respectively. A distinct deviation in the maturity stages, GSI as well as gonadal hormones (testosterone, estradiol 17-β and vitellogenin) was further observed to change significantly in specimens of African catfish captured from the Ganga River as compared to those available in culture. Highest level of testosterone was found in males having gonadal stage V which was 184.82±10.4 pg/ml in culture and 204.82±21.34 pg/ml in river-captured specimens. The mean value of serum estradiol-17β was lowest (67.25±11.4 pg/ml) in gonadal maturity stage I and highest (328.73±24.5 pg/ml) in stage V in the river-captured C. gariepinus. The vitellogenin level in female C. gariepinus was detected in stage III, and it was maximum in stage V where it was 16.68±2.98 pg/ml in river-captured specimens and 12.63±2.12 pg/ml in cultured fish.ConclusionThe results of this study on morpho-meristic and gonadal plasticity provide first evidence of invasion success of the African catfish gravitated in the Ganga River which has now adapted to the river environments for breeding and establishing. The variations concerning different reproductive phases and the gonadal hormones in culture and river-caught C. gariepinus have been considered to contribute to the success of the colonisation and establishment. The knowledge generated on the phenotypic and reproductive plasticity of African catfish available in the Ganga River will help management and control programmes.

Effects of 2,4-dichlorophenol exposure on zebrafish: Implications for the sex hormone synthesis.

2,4-Dichlorophenol (2,4-DCP), an estrogenic endocrine disruptor, is widely spread in aquatic environments and may interfere with normal physiological functions in fish. However, the influence of this chemical on the synthesis of sex hormones is not well understood. In the present study, zebrafish (Danio rerio) were exposed to 2,4-DCP (80 and 160μg/L) with or without fadrozole (an aromatase inhibitor which inhibits the synthesis of estradiol) from 20 to 40 days post fertilization. Then, the sex ratio, the content of vitellogenin (VTG) and sex hormones (androstenedione (ASD), estrone (E1), 17β-estradiol (E2), estriol (E3), testosterone (T) and 11-ketotestosterone (11-KT)) were studied. Furthermore, the expression of genes involved in synthesis of sex hormones (cyp19a1a, cyp19a1b, 17β-hsd, 11β-hsd and cyp11b) along with the DNA methylation in cyp19a1a and cyp19a1b promoters was analyzed. The results showed that 2,4-DCP exposure led to female-biased ratio, increased the content of ASD, E2 and VTG, as well as the ratio of E2/11-KT, while decreased the levels of androgens (T and 11-KT). The sex hormonal change can be explained by the significant up-regulation of cyp19a1a, cyp19a1b, 17β-hsd and 11β-hsd genes. In addition, hypomethylation of cyp19a1a promoter was involved in this process. Notably, fadrozole can partly attenuate 2,4-DCP-induced feminization, and recover the levels of ASD, E2 and 11-KT. Thus, these results demonstrate that 2,4-DCP induces feminization in fish by disrupting the synthesis of sex hormones.

Fitness Costs of Chlorantraniliprole Resistance Related to the SeNPF Overexpression in the Spodoptera exigua (Lepidoptera: Noctuidae).

Spodopteraexigua, a multifeeding insect pest, has developed a high level of resistance to chlorantraniliprole, which is a benzoylurea insecticide that targets the ryanodine receptors (RyRs). Herein, the resistant strain (SE-Sel) and sensitive strain (SE-Sus) were obtained by bidirectional screening for six generations. The potential oviposited eggs and oviposition rate of the SE-Sel strain were dramatically lower than those of the SE-Sus strain; on the contrary, the weights of prepupae and preadult were significantly increased. As a post-mating response, the higher number of non-oviposited eggs in the SE-Sel strain was caused by a lower mating rate. In addition, the expression levels of vitellogenin (SeVg) and its receptor (SeVgR) in the SE-Sel strain were consistently lower than those in the SE-Sus strain. An RyRI4743M mutation, contributing to the resistance to chlorantraniliprole, was located in the S3 transmembrane segments and might have affected the release of calcium ions; it led to the upregulated expression of the neuropeptide SeNPF and its receptor SeNPFR, and the mating and oviposition rate were significantly recovered when the SeNPF was knocked down though RNA interference (RNAi) in the male adult of the SE-Sel strain. Moreover, the expression of the juvenile hormone-binding proteins SeJHBWDS3 and SeJHBAN in the male adult of the SE-Sel strain was significantly decreased, which proved the existence of a fitness cost from another angle. Therefore, these results indicate that the fitness cost accompanied by chlorantraniliprole resistance in S. exigua may be related to the decrease in mating desire due to SeNPF overexpression.

Effects of Wastewater Treatment Plant Effluent in a Receiving Stream on Reproductive Behavior of Fathead Minnows (Pimephales promelas)

Wastewater treatment plant effluents contain a variety of endocrine disrupting chemicals (EDCs), including chemicals with estrogenic activity such as 17β-estradiol (E2), 17α-ethinyl estradiol (EE2), and nonylphenols. These substances can affect both behavior and physiology in vertebrate animals. To explore the presence and effects of these EDCs in a natural setting, juvenile and adult male fathead minnows, Pimephales promelas, were held in cages upstream and downstream of the effluent site of a wastewater treatment plant for 21 days and subsequently tested for changes in reproductive behaviors and production of vitellogenin. Additionally, estrogenic activity in the stream was measured using a yeast bioassay. Estrogenicity was found to be significantly higher downstream of the wastewater effluent when compared to levels upstream. Vitellogenin levels did not show a correlational pattern with levels of estrogenicity in the water, but two measures of reproductive behaviors occurred significantly less often in downstream males than upstream males. This suggests that a brief (three-week) exposure to stream water containing wastewater treatment plant effluent can bring about changes in reproductive behavior of fish and that behavior may be more sensitive to low levels of environmental endocrine disruptors than vitellogenin production.

Evaluation of the effect of carbamazepine on the concentration of vitellogenin in Pseudoplatystoma magdaleniatum

Abstract Carbamazepine is a pharmaceutical used in patients with seizures and bipolar disorder, which has been found in wastewater and many water resources. This is due to the inadequate disposal of pharmaceutical waste and the lack of treatment of municipal wastewater, as is the case in Colombia. The two main hydrographic basins of Colombia are the Cauca and Magdalena rivers, which are inhabited by the endemic species Striped Catfish (Pseudoplatystoma magdaleniatum). This has become an endangered species for various reasons, including the high contamination level of these rivers. In 2019, mature adult P. magdaleniatum of both sexes were caught in the Cauca river in Colombia. This was in order to assess the concentration of vitellogenin, as a biomarker of endocrine disruption, resulting from exposure to different levels of concentration of the emerging contaminant carbamazepine for 4 months. These tests were carried out in a fish farm. A significant decrease in the vitellogenin concentration was verified in females at concentrations of 25 µg l-1 and 50 µg l-1, and in males at 50 µg l-1 of carbamazepine, with a significance level of p˂0.05. Carbamazepine could cause a negative feedback in gonadotropin secretion, acting as an estrogen mimicker that causes a decrease in the level of vitellogenin.

The Effect of Infertility on the Liver Structure, Endocrinology, and Gene Network in Japanese Flounder.

Simple SummaryInfertility is a serious disorder that is characterized by the absence of fecundity in an individual. Not only does infertility in human beings affect reproduction, it also impacts hormonal secretion, metabolism, and the immune system. The doubled haploid (DH) that is induced by artificial mitogynogenesis or androgenesis in fish often results in infertility. The causes of infertility and its effects on other organs in the DH Japanese flounder (Paralichthys olivaceus) have not yet been established. Due to many aspects, the reproductive biology of this species is largely unknown or incomplete. Here, we compared the liver structure and hormone levels between fertile and infertile Japanese flounders, constructed ceRNA networks, and performed the integrated analysis of transcriptomics and proteomics. We concluded that gonadal infertility is associated with not only changes in histological structure and hormone secretion, but also changes in metabolism, immunity, and signal transduction networks in the liver. We also identified and characterized the differently expressed non-protein coding transcripts that are involved in liver functions in response to infertility. This study expanded our understanding of the molecular mechanisms of infertility on fish.The liver can synthesize vitellogenin, the precursor of vitellin, which is needed for oocyte development and maturation. Here, we investigated the effects of infertility on liver structure, hormone regulation, and gene and protein networks in Japanese flounder (Paralichthys olivaceus). Results showed that the liver of infertile fish had fewer vacuoles and significantly lower serum vitellogenin (VTG) level than in liver of fertile fish. Whole transcriptomics analysis between infertile and fertile groups identified 2076 significantly differentially expressed (DE) mRNAs, 431 DE lncRNAs, 265 DE circRNAs, and 53 DE miRNAs. Proteomics analysis identified 838 DE proteins. Integrated analysis of whole transcriptomics and proteomics revealed 60 significantly DE genes and proteins associated with metabolism, immunity, signal transduction, and steroid biosynthesis. Moreover, non-coding RNA (miRNAs, circRNA, and lncRNA) transcripts involved in metabolism, immunity, and signal transduction in infertile liver were identified. In conclusion, this study shows that gonadal infertility is associated with not only changes in histological structure and hormone secretion but also changes in metabolism, immunity, and signal transduction networks in the liver. These results provide valuable information concerning the mechanism underlying infertility in fish.

Reduced Vitellogenesis and Female Fertility in Gper Knockout Zebrafish.

The role G-protein coupled estrogen receptor (GPER) plays in vertebrate reproduction remains controversial. To investigate GPER’s reproductive role, we generated a gper zebrafish mutant line (gper−/−) using TALENs. Gper mutant females exhibited reduced fertility with a 40.85% decrease in embryo production which was associated with a significant decrease in the number of Stage V (730–750 μm) ovulated oocytes. Correspondingly, the number of early vitellogenic follicles (Stage III, 400–450 µm) in gper−/− ovaries was greater than that in wildtypes (wt), suggesting that subsequent follicle development was retarded in the gper−/− fish. Moreover, plasma vitellogenin levels were decreased in gper−/− females, and epidermal growth factor receptor (Egfr) expression was lower in Stage III vitellogenic oocytes than in wt counterparts. However, hepatic nuclear estrogen receptor levels were not altered, and estrogen levels were elevated in ovarian follicles. These results suggest that Gper is involved in the control of ovarian follicle development via regulation of vitellogenesis and Egfr expression in zebrafish.

The effects of bisphenol A on liver proteome and mucus vitellogenin in comparison to plasma as a non-invasive biomarker in immature Siberian sturgeons (Acipenser baerii)

This study was done to evaluate the effects of Bisphenol A (BPA) on Siberian sturgeon (Acipenser baerii). As liver is the main organ in the homeostatic adjustments to stress, we used a proteomics method to address molecular response in this tissue. Also, we compared the levels of vitellogenin in plasma and mucus to propose that the last one be a non-invasive method to analyze this biomarker. The fish received 1, 10, and 100 μg g−1 week−1 BPA intraperitoneally for two weeks. The samples were taken on days 0, 7, and 14. Plasma vitellogenin level increased as the highest value was recorded in the group with 100 μg g−1 week−1 of BPA. Changes in the mucus and blood vitellogenin showed a similar pattern, suggesting that mucus could be used for evaluating the changes in blood vitellogenin. Comparative proteomics was used to determine the proteome of the liver of A. baerii in the highest dose of BPA in comparison with the control. Sixteen proteins were identified that their expression changed at least twice between the studied groups. The proteomic results showed that BPA increased the expression of proteins involved in the detoxification and metabolism, activated glycolysis, and produced necrosis in the liver.

A De Novo Transcriptomics Approach Reveals Genes Involved in Thrips Tabaci Resistance to Spinosad.

Simple SummaryInsect pest resistance to pesticides is a major problem that limits efficient management and thus decreases productivity for farmers and increases the use of harmful materials that contaminate the environment and endanger humans and beneficial organisms. A major approach for resistance management is understanding how insect pest field populations develop resistance at biological and molecular levels. In this manuscript, we studied the molecular and biological basis of resistance among tobacco thrips “onion thrips” resistant the susceptible populations to spinosad, a major insecticide that has been extensively used in recent years, by using de novo transcriptomics and bioassays. We found that resistance to spinosad can be metabolic by increasing the levels of detoxifying enzymes among the resistant population; however, resistant populations are more fecund compared to susceptible one, suggesting the lack of fitness cost of the resistance trait.The onion thrip, Thrips tabaci (Thysanoptera: Thripidae) is a major polyphagous pest that attacks a wide range of economically important crops, especially Allium species. The thrip’s damage can result in yield loss of up to 60% in onions (Allium cepa). In the past few decades, thrip resistance to insecticides with various modes of actions have been documented. These include resistance to spinosad, a major active compound used against thrips, which was reported from Israel. Little is known about the molecular mechanisms underlying spinosad resistance in T. tabaci. We attempted to characterize the mechanisms involved in resistance to spinosad using quantitative transcriptomics. Susceptible (LC50 = 0.6 ppm) and resistant (LC50 = 23,258 ppm) thrip populations were collected from Israel. An additional resistant population (LC50 = 117 ppm) was selected in the laboratory from the susceptible population. De novo transcriptome analysis on the resistant and susceptible population was conducted to identify differently expressed genes (DGEs) that might be involved in the resistance against spinosad. In this analysis, 25,552 unigenes were sequenced, assembled, and functionally annotated, and more than 1500 DGEs were identified. The expression levels of candidate genes, which included cytochrome P450 and vittelogenin, were validated using quantitative RT-PCR. The cytochrome P450 expression gradually increased with the increase of the resistance. Higher expression levels of vitellogenin in the resistant populations were correlated with higher fecundity, suggesting a positive effect of the resistance on resistant populations. This research provides a novel genetic resource for onion thrips and a comprehensive molecular examination of resistant populations to spinosad. Those resources are important for future studies concerning thrips and resistance in insect pests regarding agriculture.

In vivo and in silico analyses of estrogenic potential of equine estrogens in medaka (Oryzias latipes)

Equine estrogens (EEs) are widely used in hormone replacement therapy pharmaceuticals for postmenopausal women. Previous studies have shown that EEs occur in the aquatic environment; however, the potential estrogenicity and risk of EEs in aquatic organisms, including fish, have yet to be studied in detail. Therefore, we evaluated the estrogenic potential of major EEs, namely equilin (Eq), 17α-dihydroequilin (17α-Eq), 17β-dihydroequilin (17β-Eq), equilenin (Eqn), 17α-dihydroequilenin (17α-Eqn), and 17β-dihydroequilenin (17β-Eqn), on medaka (Oryzias latipes) using in vivo and in silico assays. Quantitative real-time RT-PCR analyses revealed that expression levels of choriogenin L (ChgL) and choriogenin H (ChgH) in medaka embryos responded to various types and concentrations of EEs in a concentration-dependent manner, whereas transcription levels of vitellogenin 1 were not significantly affected by any of the EEs in the concentration range tested. The order of the in vivo estrogenic potencies of EEs was as follows: 17β-Eq > Eq > 17β-Eqn > Eqn > 17α-Eqn > 17α-Eq. Additionally, the 50% effective concentrations (EC50) of 17β-Eq was lower than that of 17β-estradiol. We also investigated the interaction potential of EEs with medaka estrogen receptor (ER) subtypes in silico using a three-dimensional model of the ligand-binding domain (LBD) for each ER and docking simulations. All six EEs were found to interact with the LBDs of ERα, ERβ1, and ERβ2. The order of the in silico interaction potentials of EEs with each ER LBD was as follows: 17β-Eq > 17α-Eq > Eq > 17β-Eqn > 17α-Eqn > Eqn. Furthermore, we identified the key amino acids that interact with EEs in each ER LBD; our findings suggest that amino acids and/or their hydrogen bonding may be responsible for the ligand-specific interactions with each ER. This study is the first to comprehensively analyze the estrogenic potential of EEs in medaka both in vivo and in silico.

17\u03b1-ethynylestradiol prevents the natural male-to-female sex change in gilthead seabream (Sparus aurata L.)

Exposure to 17α-ethynylestradiol (EE2, 5 μg/g food) impairs some reproductive events in the protandrous gilthead seabream and a short recovery period does not allow full recovery. In this study, spermiating seabream males in the second reproductive cycle (RC) were fed a diet containing 5 or 2.5 μg EE2/g food for 28 days and then a commercial diet without EE2 for the remaining RC. Individuals were sampled at the end of the EE2 treatment and then at the end of the RC and at the beginning of the third RC, 146 and 333 days after the cessation of treatment, respectively. Increased hepatic transcript levels of the gene coding for vitellogenin (vtg) and plasma levels of Vtg indicated both concentrations of EE2 caused endocrine disruption. Modifications in the histological organization of the testis, germ cell proliferation, plasma levels of the sex steroids and pituitary expression levels of the genes coding for the gonadotropin β-subunits, fshβ and lhβ were detected. The plasma levels of Vtg and most of the reproductive parameters were restored 146 days after treatments. However, although 50% of the control fish underwent sex reversal as expected at the third RC, male-to female sex change was prevented by both EE2 concentrations.

2,4,6-Tribromophenol is toxic to Oreochromis niloticus (Linnaeus, 1758) after trophic and subchronic exposure

The presence of 2,4,6-Tribromophenol (TBP) in the environment increased the risk of exposure to aquatic organisms affecting the animal development or metabolism. The current study investigated the low, subchronic and trophic effect of TBP in both, male and female adult of Oreochromis niloticus. The fish were exposed to 0.5 or 50 ng g−1 of TBP every ten days for 70 days. Then, hepatosomatic (HSI) and gonadosomatic (GSI) indexes, erythrocyte parameters (hemoglobin content, nuclear morphology and morphometrical abnormalities), biochemical endpoints (glutathione S-Transferase and catalase activities, non-protein thiols, lipid peroxidation and protein carbonylation levels in the liver; and acetylcholinesterase activity in the brain and muscle), histopathological analysis (liver) and vitellogenin levels (plasma) were considered. TBP affected the HSI in male and female fish, but not the GSI. Principal Component Analysis revealed that erythrocytes from males are more sensitive to TBP exposure. Likewise, TBP induced the expression of vitellogenin, CAT activity and liver lesion in male fish comparatively with control group, but GST and NPT were influenced only by sex. Finally, the results showed that the antioxidant mechanism and cholinesterase activity effects were more pronounced in male than in female. The current data shows evidences of estrogenic endocrine disruption and toxicity in O. niloticus exposed to TBP, revealing the risk of exposure to biota.

Early gonadal maturation and vitellogenin mRNA expression in Siberian sturgeon Acipenser baerii cultured in a semi-closed water recirculating system in Korea

Changes of gonadal morphology and mRNA expression patterns of vitellogenin were investigated in Siberian sturgeon Acipenser baerii (Chondrostei) during its early gonadal maturation period. Early differentiations and morphological transitions of both ovaries and testes appeared to occur actively until the age of 3 years, however from then on, the maturation patterns to full maturity were largely gender-dependent, in which males showed a faster progression of maturation than did females while females experienced a steady-state progress with a lagged interval before entering the final maturation. Expression of vitellogenin mRNAs are closely correlated with transitional patterns of gonadal appearances. In both females and males, hepatic mRNA levels of vitellogenin exponentially increased in the earliest interval (up to 1-year-old). However, in subsequent periods, vitellogenin expression in females continued to increase with age, whereas in males, the expression stabilized at a younger age. Nevertheless, at the age older than or equal to 7-year-old, fully matured individuals showed a quite low level of vitellogenin expression in both females and males. Collectively, results from this study could be useful as a fundamental guideline to address the gonad maturation of this sturgeon species, which is helpful for making practical decisions about farming practices and management for caviar production on local sturgeon farms.