The probiotic SLAB51 as agent to counteract BPA toxicity on zebrafish gut microbiota -liver-brain axis.

A plethora of studies have so far described the toxic effects of bisphenol A (BPA) on organism health, highlighting the urgent need to find new strategies not only to reduce the presence of this toxicant but also to counteract its adverse effects. In this context, probiotics emerged as a potential tool since they promote organism welfare. Using a multidisciplinary approach, this study explores the effects of SLAB51 dietary administration to counteract BPA toxicity using zebrafish as a model. Adult males and females were maintained under standard conditions (control group; C), exposed for 28 days via the water to an environmental relevant dose of BPA (10 µg/L; BPA), dietary treated with SLAB51 (109 CFU/g of body weight; P) and co-treated with BPA plus SLAB51 (BPA + P). In the gut, exposure to BPA resulted in altered architecture in both males and females, with females also experiencing an increase of pathogenic bacterial species. Co-administration of BPA + P led to the restoration of normal gut architecture, favored beneficial bacteria colonization, and decreased the abundance of pathogenic species. In the liver, male BPA exposure led to steatosis and glycogen depletion, which was partially mitigated by SLAB51 co-administration. In contrast, in females exposed to BPA, the lack of steatosis along with the greater glycogen depletion, suggested an increase in energy demand as supported by the metabolomic phenotype. The analysis of liver metabolites in BPA + P males revealed increased levels of anserine and reduced levels of glutamine, which could lie behind the counteraction of the brain histopathological damage caused by BPA. In BPA + P females, a reduction of retinoic acid was found in the liver, suggesting an increase in retinoids responsible for BPA detoxification. Overall, these results demonstrate that SLAB51 exerts its beneficial effects on the gut microbiota-brain-liver axis through distinct molecular pathways, effectively mitigating the pleiotropic toxicity of BPA.

Metabolomic and Transcript Analysis Revealed a Sex-Specific Effect of Glyphosate in Zebrafish Liver.

Glyphosate is a component of commonly used herbicides for controlling weeds in crops, gardens and municipal parks. There is increasing awareness that glyphosate-based herbicides, in addition to acting on plants, may also exert toxicity in wildlife and humans. In this study, male and female adult zebrafish were exposed to 700 µg/L of glyphosate (GLY), for 28 days. We used the metabolomic approach and UHPLC-ESI-MS to analyze liver samples to investigate the adverse effects of glyphosate on hepatic metabolism. The impact of GLY was found to be sex-specific. In female, GLY exposure affected purine metabolism by decreasing the levels of AMP, GMP and inosinic acid, consequently increasing uric acid levels with respect to the control (CTRL). Exposure to GLY also caused a decrease of UMP levels in the pyrimidine metabolism pathway. In male, GLY exposure decreased the aminoadipic acid within the lysine degradation pathway. Transcript analysis of genes involved in stress response, oxidative stress and the immune system were also performed. Results demonstrated an increased stress response in both sexes, as suggested by higher nr3c1 expression. However, the hsp70.2 transcript level was increased in female but decreased in male. The results demonstrated reduced sod1, sod2, and gpx1a in male following exposure to GLY, indicating an impaired oxidative stress response. At the same time, an increase in the cat transcript level in female was observed. mRNA levels of the pro-inflammatory interleukins litaf and cxcl8b.1 were increased in female. Taken together, the results provide evidence of disrupted nucleotide hepatic metabolism, increased stress inflammatory response in female and disruption of oxidative stress response in male.

Metabolic Changes During Growth and Reproductive Phases in the Liver of Female Goldfish (Carassius auratus).

Hormones of the brain-pituitary-peripheral axis regulate metabolism, gonadal maturation, and growth in vertebrates. In fish, reproduction requires a significant energy investment to metabolically support the production of hundreds of eggs and billions of sperms in females and males, respectively. This study used an LC-MS-based metabolomics approach to investigate seasonally-related changes in metabolic profile and energy allocation patterns in female goldfish liver. We measured basal metabolic profile in female goldfish at three phases of the reproductive cycle, including 1) Maximum growth period in postovulatory regressed phase, 2) mid recrudescence in fish with developing follicles, and 3) late recrudescence when the ovary contains mature ovulatory follicles. We also investigated changes in the liver metabolism following acute treatments with GnRH and GnIH, known to be involved in controlling reproduction and growth in goldfish. Chemometrics combined with pathway-driven bioinformatics revealed significant changes in the basal and GnRH/GnIH-induced hepatic metabolic profile, indicating that metabolic energy allocation is regulated to support gonadal development and growth at different reproductive cycles. Overall, the findings support the hypothesis that hormonal control of reproduction involves accompanying metabolic changes to energetically support gonadotropic and somatotropic activities in goldfish and other oviparous vertebrates.

Seasonally related metabolic changes and energy allocation associated with growth and reproductive phases in the liver of male goldfish (Carassius auratus).

Reproduction and growth follow a seasonal pattern in many fish species involving changes in gonadal development, growth, and metabolism. Significant metabolic energy is needed during gametogenesis in both female and male to produce hundreds of eggs and billions of sperms. Seasonal variations are controlled by the hormones of brain-pituitary-peripheral axis and are accompanied by significant metabolic changes. There is evidence that GnRH and GnIH are among the key neurohormones that regulate the reciprocal control of growth and reproduction. The objective of this study was to investigate changes in metabolic profile and energy allocation patterns at different stages of reproduction, using goldfish as a model organism and LC-MS as analytical platform for metabolic analysis. Goldfish undergoes a clear seasonal cycle of growth and reproduction. In vivo experiments were conducted at three different time point of the annual cycle: regressed gonadal phase (peak growth phase), mid gametogenesis and late gametogenesis. Emphasis is placed on changes in liver metabolic pathways to energetically sustain the physiological processes related to growth and reproduction. Moreover, we tested the hypothesis that GnRH and GnIH may play a role in the regulation of metabolism by investigating acute effects of these peptides at different stages of reproductive cycle. SIGNIFICANCE: The findings in this paper provide novel information on the seasonal changes in basal metabolism during different stages of reproductive cycle, and evidence for differential allocation of energy during reciprocal control of reproduction and growth in goldfish. Chemometrics combined with pathway-driven bioinformatics elucidated a shift in the metabolic profile, indicating distinct patterns of energy allocation in the reproductive and growth seasons. Furthermore, to our knowledge this is the first study to provide evidence for a possible regulatory role of GnRH and GnIH in liver metabolism and energy allocation patterns associated with growth and reproductive processes. Together our findings present a framework for better understanding of the hormonally induced changes in metabolism to energetically sustain growth and reproduction in fish and other oviparous species undergoing seasonal cycle.

Seasonal Related Multifactorial Control of Pituitary Gonadotropin and Growth Hormone in Female Goldfish: Influences of Neuropeptides and Thyroid Hormone.

Female reproduction is under multifactorial control of brain-pituitary-peripheral origin. The present study provides information on seasonal changes in circulating LH and GH concentrations, as well as transcript levels for a number of genes involved in the regulation of reproduction and growth in female goldfish. We also provide information on the effects of treatments with GnRH and/or GnIH, and their interaction with T3, at three stages of gonadal recrudescence. Maximum basal concentration of LH was observed at late recrudescence (Spring) while no seasonal changes in basal serum GH levels was detected. Serum LH and GH levels were stimulated by GnRH as expected, depending on the season. GnIH stimulated basal GH concentrations in gonadally regressed fish. GnIH inhibitory action on GnRH-induced LH response was observed in late, but not in mid recrudescence. T3 actions on basal and GnRH- or GnIH-induced GH secretion were generally inhibitory, depending on season. Administration of T3 attenuated GnRH-induced LH responses in mid and late stages of gonadal recrudescence, and the presence of GnIH abolished inhibitory actions of T3 in fish at mid recrudescence. Our results also demonstrated seasonal patterns in basal and GnRH- and/or GnIH-induced transcript levels for ERα, ERßI, FSHR, aromatase, TRαI, TRß, IGF-I, and Vtg in the liver and ovary. However, there were no clear correlations between changes in transcript levels and circulating levels of LH and GH. The results support the hypothesis that GnRH, GnIH, and T3 are contributing factors in complex reciprocal control of reproduction and growth in goldfish.

In-vitro and in-vivo biological activity of recombinant yellowtail kingfish (Seriola lalandi) follicle stimulating hormone.

Biologically active recombinant yellowtail kingfish follicle stimulating hormone (rytkFsh) was produced in yeast Pichia pastoris and its biological activity was demonstrated by both in-vitro and in-vivo bioassays. Incubation of ovarian and testicular fragments with the recombinant hormone stimulated E2 and 11-KT secretion, respectively. In-vivo trial in immature female YTK resulted in a significant increase of plasma E2 levels and development of oocytes. In males at the early stages of puberty, advancement of spermatogenesis was observed, however plasma 11-KT levels were reduced when administered with rytkFsh.