•Diets contaminated with NP, BPA, or t-OP affect lipid metabolism.•Xenobiotic-contaminated diets induce metabolic disorders.•Hepatic metabolic disorders may be related to environmental pollution.
The ...metabolic effects induced by feed contaminated with a lower or a higher concentration of -nonylpnenol (NP), 4-tert-octylphenol (t-OP) or bisphenol A (BPA), three environmental endocrine disruptors, were assessed in juvenile sea bream liver. Histological analysis demonstrated that all these three xenobiotics induced hepatic lipid accumulation and steatosis. These findings prompted analysis of the expression of the major molecules involved in lipid metabolism: peroxisome proliferator activated receptors (which is encoded by ppars), fatty acid synthase (encoded by fas), lipoprotein lipase (encoded by lpl) and hormone-sensitive lipase (encoded by hsl). The enzymes encoded by ppars and fas are in fact responsible for lipid accumulation, whereas lpl- and hsl- encoded proteins play a pivotal role in fat mobilization. The three xenobiotics modulated ppar mRNA expression: pparα mRNA expression was induced by the higher dose of each contaminant; pparβ mRNA expression was upregulated by the lower doses and in BPA2 fish ppary mRNA overexpression was induced by all pollutants. These data agreed with the lipid accumulation profiles documented by histology. Fas mRNA levels were modulated by the two NP doses and the higher BPA concentration. Lpl mRNA was significantly upregulated in all experimental groups except for BPA1 fish while hsl mRNA was significantly downregulated in all groups except for t-OP2 and BPA1 fish. The plasma concentrations of cortisol, the primary stress biomarker, were correlated with the levels of pepck mRNA level. This gene encodes phosphoenolpyruvate carboxykinase which is one of the key enzymes of gluconeogenesis. Pepck mRNA was significantly overexpressed in fish exposed to NP2 and both t-OP doses. Finally, the genes encoding cyclooxygenase 2 (cox2) and 5-lipoxygenase (5 lox), the products of which are involved in the inflammatory response, transcriptions were significantly upregulated in NP and BPA fish, whereas they were unchanged in t-OP specimens. The present findings suggest that dietary xenobiotic contamination can give rise to metabolic disorders also in fish and highlight the potential for their vertical transfer through the trophic levels and ultimately to humans.
•Diets contaminated with a mixture of xenoestrogens affected lipid metabolism.•Environmental pollution is a causal factor of hepatic steatosis.•FT-IR analysis showed the increase of lipids in hepatic ...tissue.•LC/ESI-QTRAP-MS/MS analysis showed the ability of NP to accumulate in the muscle of fish.
Many man-made chemical compounds are recognized as endocrine disruptors and once released into the environment are likely to spread and bioaccumulate in wild species. Due to their lipophilic nature, these substances pass through the cell membrane or bind to specific receptors activating physiological responses that in the long run can cause reproductive impairment, physiological disorders, including the occurrence of metabolic syndromes. One significant source of contamination is represented by the consumption of polluted food. As a consequence, different environmental pollutants, with similar or different modes of action, can accumulate in organisms and biomagnify along the food web, finally targeting humans. The aim of this study was to analyze, under controlled conditions, the effects induced by the consumption of contaminated diets, focusing on the effects exerted at hepatic level. Juvenile seabream were fed for 21days a diet enriched with different combinations of pollutants, nonylphenol (NP), tert-octylphenol (t-OP) and bisphenol A (BPA). The different diets containing 5mg/kg bw of each contaminant, were formulated as follows: NP+tOP, BPA+NP, BPA+tOP and NP+BPA+tOP (NBO). EDCs, at the doses administered, showed low biomagnification factor (BMF), suggesting that these pollutants hardly accumulate in muscles. The results obtained at hepatic level pinpointed the steatotic effect of all the administered diets, associated to a modulation of the expression of genes involved in lipid metabolism (ppars, fas, lpl, and hsl). Results were compared to those obtained in previous studies in which fish were fed single pollutants evidencing that the administration of mixture of contaminants exerts a milder lipogenic effect, highlighting the contrasting/antagonistic interaction establishing among chemicals. Noteworthy was the setup of a new chromatographic method to detect the presence of the selected chemical in fish muscle and the application of Fourier Transform Infrared (FT-IR) analysis to evaluate pollutant-induced changes in the liver macromolecular building.
Several complex processes are involved in the production of viable eggs. The aim of this review is to provide an overview on the role played by lysosomal enzymes, especially cathepsins B, D, and L, ...during ovarian follicle growth and maturation. Specific attention is focused on the relationship between the second proteolytic cleavage of yolk proteins (YP) and the resumption of the meiosis during germinal vesicle break down (GVBD). Maturation represents the final stage of oocytes development prior to ovulation. Oocytes in this phase appear translucent. In many teleosts GVBD is accompanied by water uptake and among marine teleosts with pelagic eggs, most of the final volume is reached by this process. The last phase of maturation in benthonic eggs also occurs concomitant to a second proteolytic cleavage and is related with a slight hydration process. In vitro maturation by 17α,20β-dihydroxy-4-pregnen-3one in class III
Danio rerio oocytes, induced 80% of GVBD. The maturation of these oocytes is known to be associated with proteolysis of their major yolk components. In the present study, we show that inhibition of specific enzymes (cathepsins) involved in the second YP processing, did not affect the occurrence of GVBD as the oocytes become translucent and display a slight increase in size. More specifically, in vitro incubation of the maturing oocytes with a cathepsin B inhibitor suppressed both cathepsin B and L activities and the proteolysis of YP. On the contrary, the addition of cathepsin L inhibitor, only affected cathepsin L activity, indicating that cathepsin B is probably involved in Cathepsin L activation, and this enzyme is probably responsible for the second YP processing. These results, together with previous studies, indicate that the GVBD process is independent of the occurrence of the second proteolytic process. It supports the hypothesis that the maturation process is under K
+ ion flux control, while yolk proteolysis is related to the temporal and specific activation of cathepsins by acidification of yolk spheres.
The aim of this study was to evaluate the potential use of the harpacticoid copepod Tisbe spp as prey in Amphiprion clarkii larviculture. After hatching, A. clarkii larvae were divided in four ...experimental groups for feeding studies as follows: group A (control group) fed rotifers (Brachionus plicatilis) followed by Artemia nauplii; group B fed a mixed diet of rotifers and Tisbe spp nauplii followed by a combination of A. nauplii and Tisbe spp copepodites/copepods; group C fed copepod nauplii solely followed by Tisbe spp copepodites and copepods; group D fed rotifers followed by Tisbe spp copepodites and copepods. In this study we observed a positive effect of feeding Tisbe spp copepods in A. clarkii larviculture as a supplement live food to the traditional diet based on rotifers and A. nauplii. In group B larvae, fed a combination of rotifers/Tisbe spp nauplii followed by a combination of A. nauplii/Tisbe spp copepodites-adults, a significant increase of IGF II and IGF I gene expression and a significant decrease of myostatin gene expression was evidenced by Real Time PCR compared to the other experimental groups. In this same group we also observed the best results in terms of growth (total length and weight) and survival.
In conclusion, the harpacticoid copepod Tisbe spp may be considered a suitable live prey for marine fish larvae larviculture when used as a supplement to the traditional diet based on rotifers and A. nauplii.
Research highlights ► EDs impair body weight. ► In the hypothalamus several peptidergic hormones and neurotransmitters controlling feeding behaviour are impaired by EDs. ► EDs derange lipid ...metabolism by targeting key metabolic sensors. ► The phthalate DEHP at low doses triggers lipid synthesis and decreases food intake stimulus.
A multi‐technique approach was used to study the changes occurring in European eel Anguilla anguilla ovaries during hormonally‐induced vitellogenesis. Aside from classic techniques used to monitor ...the vitellogenic process, such as ovary histology, fat content analysis, sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and vitellogenin enzyme linked immunosorbent assay (ELISA), a new technique, Fourier‐transform infrared (FT‐IR) microspectroscopy, was used to analyse A. anguilla ovaries. The results from the different techniques provided different ways of approaching the same process. Although it is considered a time consuming approach, of all the employed techniques, histology provided the most direct evidences about vitellogenesis. SDS–PAGE and ELISA were also useful for studying vitellogenesis, whereas fat analysis cannot be used for this purpose. The FT‐IR analysis provided a representative IR spectrum for each ovarian stage (previtellogenic stage, early vitellogenic stage, mid‐vitellogenic stage and late vitellogenic stage), demonstrating that it is a valid method able to illustrate the distribution of the oocytes within the ovary slices. The chemical maps obtained confirmed changes in lipid concentrations and revealed their distribution within the oocytes at different maturational stages. When the results and the accuracy of the FT‐IR analysis were compared with those of the traditional techniques commonly used to establish the vitellogenic stage, it became evident that FT‐IR is a useful and reliable tool, with many advantages, including the fact that it requires little biological material, the costs involved are low, analysis times are short and last but not least, the fact that it offers the possibility of simultaneously analysing various biocomponents of the same oocyte.
At present, clownfishes are the best example of successfully captive bred ornamental specimens but little is known about the relationship between food enrichment and both larval growth and ...development. In fact, it is well known that a certain percentage of these fishes cultured in captive conditions show a miss-band pigmentation. In the present study, the effects of live prey enrichment on growth and pigmentation in false percula clownfish (Amphiprion ocellaris) larvae were tested.
Newly hatched A. ocellaris larvae were divided in three different groups and fed as follows: group A fed on enriched (Algamac 2000) B. plicatilis (10 ind./mL) from day 1 to day 5 post-hatch (ph); group B fed on enriched (Algamac 2000) B. plicatilis (10 ind./mL) followed by Artemia nauplii (5 ind./mL;) and group C fed on Algamac 2000 enriched B. plicatilis and Algamac 2000 enriched Artemia nauplii.
Samples of the larvae were collected on day 5 from group A and on day 11 ph in group B and C for morphometric and molecular analysis. On day 11 ph food enrichment resulted in a better growth of group C larvae respect to those of group B fed on not enriched Artemia nauplii (15.8±0.2 mg and 8.78±0.02 mm vs. 6.8±0.2 mg and 6.93±0.01 mm). Moreover, 36±2% of the juveniles obtained from group B showed a miss-band pigmentation as compared to 29±1% of the juveniles obtained from group C. At molecular level, the results obtained by Real-Time PCR are in agreement with the morphometric ones: a positive induction of the Insulin-like Growth Factor II (IGFII) and Peroxisome Proliferator Activated Receptor α and β (PPARα and PPARβ) gene expression and a reduction of Myostatin (MSTN) was observed in group C larvae fed on enriched live prey. IGFI gene expression was higher in group B.
The present study provides clear evidences of the positive role of Algamac 2000 on growth and pigmentation of captive cultured false percula clownfish.
It is known that heavy metals can accumulate in tissues during aquatic organism growth (bioaccumulation) and often biomagnify up the food chain interfering with the health and reproduction of both ...wildlife and humans. Recently, cadmium (Cd) was included in the endocrine disruptors list, exerting its effect on gametes quality and reproductive functions; in addition, its role as apoptotic factor was evidenced in different cell types and tissues. In the present study, the effects of two different Cd doses on testis and liver of the black goby
Gobius niger were analyzed. Cd concentration in the water and its uptake by the gills were measured by differential pulse anodic stripping voltammetry. Toxic, apoptotic, and stressor Cd effects were analyzed using metallothionein (MTT), caspase 3 and heath shock protein 70 (HSP70), respectively, as bioindicators. The results of the present study suggested that, in the gills, the saturation of all specific metal sites was reached only with the highest Cd dose exposure. Either testis and liver showed an increase of MTT gene expression and protein synthesis in addition to HSP70 gene expression, related with Cd concentration in the water indicating that both tissues were affected by Cd exposure. In conclusion, the present study, not only shows the toxic effect of Cd on hepatic tissue, but also indicates its potency as apoptotic factor in the testis. This is supported by the increase of caspase 3 gene expression and the presence of its active form in testis of exposed fish.
In the present study, a cDNA for the hatching enzyme of a marine tropical fish, Chysiptera parasema, was cloned. This is the first demonstration of hatching enzyme cDNA from a marine tropical fish. ...The amino acid (aa) sequence deduced from the cDNA consisted of an 18-aa signal sequence, a 53-aa propeptide sequence and a 196-aa mature enzyme portion, having a consensus active site sequence for astacin family proteases. Phylogenetic analysis showed that the C. parasema enzyme was included in the clade of HCEs (high choriolytic enzymes), one of the hatching enzymes of freshwater fishes such as medaka (Oryzias latipes), masu salmon (Oncorhynchus masou) and zebrafish (Danio rerio), but not in the group of LCEs (low choriolytic enzymes), another type of hatching enzymes identified in the medaka. The developmental expression patterns of the C. parasema HCE gene were highly similar to that of the medaka HCE gene. The results suggested that the hatching enzyme system is highly conserved between these marine and freshwater fish species. PUBLICATION ABSTRACT
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