Publications by authors named "Giovana Souza Branco"

6 Publications

  • Page 1 of 1

Nonsteroidal anti-inflammatory drugs act as endocrine disruptors in Astyanax lacustris (Teleostei: Characidae) reproduction: An ex vivo approach.

Aquat Toxicol 2021 Jan 29;232:105767. Epub 2021 Jan 29.

Escola de Ciências da Saúde, Universidade Anhembi Morumbi - R. Dr. Almeida Lima, 1134 - Parque da Mooca, 03164-000, São Paulo, SP, Brazil. Electronic address:

Pharmaceutical products can act as endocrine-disrupting compounds (EDCs), affecting the physiological processes of animals, such as development or reproduction. This study aimed to investigate the influence of different concentrations of nonsteroidal anti-inflammatory drugs (NSAIDs) diclofenac (DCF) and ibuprofen (IBU) alone and mixed (MIX) on gonadotropin gene expression and gonadal steroid release using Astyanax lacustris pituitary and testes explant systems, respectively. The explant organs were maintained for 12 h in Leibovitz (L-15) medium supplemented with 0, 0.1, 1, 10, 100, and 1000 ng L of DCF, IBU, and MIX (ratio 1:1 of the same concentrations of DCF and IBU alone) and gonadotropin releasing-hormone (cGnRH2) stimulation in pituitary explants and human chorionic gonadotropin (hCG) stimulation in testes explants. The pituitary glands and the media from the testicular explants were collected for gene expression analysis including the β subunit of the follicle-stimulating hormone (fshβ) and luteinizing hormone (lhβ) and secreted gonadal steroid concentration analysis, respectively. Both DCF and IBU (alone and mixed) decreased pituitary gene expression of fshβ and lhβ and this inhibitory effect was evident even at low concentrations. In the testes, DCF and IBU did not change the levels of estradiol, and both pharmaceuticals increased the release of 11-ketotestosterone at low doses, while only IBU decreased the levels of testosterone in all concentrations. IBU's inhibitory effect in the testes was not triggered by the mixture of the two drugs. These results suggest that NSAIDs, may interfere in fish reproduction by acting as EDCs, thereby negatively affecting A. lacustris spermatogenesis and maturation.
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http://dx.doi.org/10.1016/j.aquatox.2021.105767DOI Listing
January 2021

Aluminum at environmental concentrations affects the sperm quality of the freshwater teleost Astyanax altiparanae: An in vitro study.

Comp Biochem Physiol C Toxicol Pharmacol 2021 May 22;243:108983. Epub 2021 Jan 22.

Universidade de São Paulo, Instituto de Biociências, Departamento de Fisiologia, Laboratory of Metabolism and Reproduction of Aquatic Organisms - LAMEROA, Rua do Matão, trav. 14, 101, Cidade Universitária, São Paulo, SP, Brazil. Electronic address:

Aluminum (Al) is present in rivers and reservoirs in concentrations above that is allowed by regulatory agencies (e.g. 0.5 mg L Al), which can impair fish reproduction. The present study evaluated the in vitro effects on the sperm of Astyanax altiparanae upon Al exposure at different concentrations (0, 0.05, 0.1, 0.3, and 0.5 mg L) with various exposure periods (50 s, 10 min, and 30 min). The following biomarkers were evaluated: membrane vitality, DNA fragmentation, morphology, kinetics (10 s and 30 s after sperm activation), and sperm mitochondrial activity. Al damages the membrane vitality of gametes at 0.3 and 0.5 mg L after 50 s of exposure. After 30 min of exposure, there was a decrease in membrane vitality at 0.1 and 0.5 mg L, and the membrane vitality decreased with increased exposure time. Within 30 s after sperm activation, Al (0.3 and 0.5 mg L) reduced sperm motility by more than 50% at the longest exposure time, while at 0.1 and 0.5 mg L, Al exposure reduced motility over time. The average path speed (VAP; 10 s post-sperm activation) was reduced at longer exposure times at 0.05 and 0.5 mg L of Al. Increased exposure time had deleterious effects on mitochondrial activity at the highest concentrations tested. Al did not damage DNA and sperm morphology. In conclusion, Al negatively influences the sperm quality of A. altiparanae with a potential effect of exposure time and increasing concentrations.
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http://dx.doi.org/10.1016/j.cbpc.2021.108983DOI Listing
May 2021

Paternal exposure to aluminum, acidity, and temperature affect fatty acid seminal profile, embryonic and larval development of Astyanax altiparanae.

Chemosphere 2021 Mar 11;266:128935. Epub 2020 Nov 11.

Universidade de São Paulo, Instituto de Biociências, Departamento de Fisiologia, Laboratory of Metabolism and Reproduction of Aquatic Organisms - LAMEROA, Matão Street, 14 Lane, Number 101, Room 220, Cidade Universitária, São Paulo, SP, Brazil. Electronic address:

We investigated the effects of water acidity, temperature, and aluminum (Al) on the fatty acid (FA) seminal profile, reproductive parameters (fertilization and hatching) and embryonic development of Astyanax altiparanae. We treated males with different experimental treatments, corresponding to the combination of water temperature (20 °C; 25 °C), pH (neutral - 7.0; acidic - 5.5), and the absence or presence of Al (0.5 mg L). After 96 h, we analyzed the FA profile of semen and performed artificial fertilization in activating medium with neutral pH or activating medium in the same experimental conditions of the males (neutral pH, acidic pH, and Al) to evaluate fertilization and hatching rates and to monitor embryonic development. Polyunsaturated FA percentage decreased in semen of fish from the neutral group, while monounsaturated FA increased in all groups maintained at 20 °C compared to 25 °C. Aluminum exposure decreased the percentage of C20:4n6 and increased the percentage of C22:5n3 at 20 °C. Males exposed to acidic pH and Al showed lower fertilization and hatching rates, as well as increased mortality of embryos and larvae. Moreover, Al favoured a higher percentage of abnormal larvae. Fertilization in Al activating medium harmed the embryos and larvae since fertilization and hatching rates decreased. Finally, temperature influenced fertilization time, hatching rate, and the morphology of embryos and larvae. Males exposed to Al had lower fertilizing capacity, which negatively affected the embryonic development of the species. Furthermore, Al activating medium reduced the number of fertilized oocytes, hatched embryos, and normal larvae. All events were temperature dependent.
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http://dx.doi.org/10.1016/j.chemosphere.2020.128935DOI Listing
March 2021

Diclofenac and caffeine inhibit hepatic antioxidant enzymes in the freshwater fish Astyanax altiparanae (Teleostei: Characiformes).

Comp Biochem Physiol C Toxicol Pharmacol 2021 Feb 10;240:108910. Epub 2020 Oct 10.

Universidade de São Paulo, Instituto de Biociências, Departamento de Fisiologia, Laboratório de Metabolismo e Reprodução de Organismos Aquáticos, Cidade Universitária, São Paulo, Brazil.

Although concentrations of pharmaceutical compounds in aquatic ecosystems are low, they can cause toxic effects on organisms. The aim of this study was to evaluate the effects of diclofenac (DCF), a non-steroidal anti-inflammatory drug, and caffeine (CAF), a central nervous system stimulant, both alone or combined, in Astyanax altiparanae males under acute exposure (96 h), measuring neurotoxicity biomarkers, antioxidant response and damage at biochemical and cellular levels. DCF concentration in water, separated and combined, was 3.08 mg L and that of CAF was 9.59 mg L. To assess neurotoxicity, brain and muscle acetylcholinesterase (AChE) activities were measured. To evaluate oxidative stress, the enzymatic activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and glutathione S-transferase (GST), as well as lipoperoxidation (LPO), were analyzed in liver and gills. Activity of hepatic cyclooxygenase (COX) was also evaluated. Genotoxicity was assessed in blood using comet assay and micronucleus test, as well as nuclear abnormalities. DCF and CAF, alone or combined, had neither effect on AChE activity, nor in the activity of SOD, CAT, GPx and GST in gills. In liver, DCF inhibited SOD and GPx activity, CAF inhibited CAT activity, the mixture inhibited SOD and GST activity; although only fish exposed to CAF showed increased hepatic LPO. Under these experimental conditions, no effect on COX activity was observed, nor cytotoxic and genotoxic damage. The most pronounced effects were caused by the drugs separately, since both compounds altered the enzymes, but only CAF triggered LPO, showing more harmful effects.
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http://dx.doi.org/10.1016/j.cbpc.2020.108910DOI Listing
February 2021

Effects of GnRH and the dual regulatory actions of GnIH in the pituitary explants and brain slices of Astyanax altiparanae males.

Gen Comp Endocrinol 2019 03 9;273:209-217. Epub 2018 Aug 9.

Reproductive and Molecular Biology Group, Department of Morphology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu Campus, Botucatu, Brazil. Electronic address:

The pituitary gonadotropins, Fsh (follicle-stimulating hormone) and Lh (luteinizing hormone), regulate testicular development and functions in all vertebrates. At the pituitary, different signaling systems regulate the synthesis and secretion of the gonadotropins, such as the hypothalamic neuropeptides GnRH (gonadotropin-releasing hormone) and GnIH (gonadotropin-inhibitory hormone). While GnRH exerts stimulatory roles, the actions of GnIH remain controversial for many teleost species. Therefore, the aim of this study was to evaluate the in vitro effects of chicken GnRH2 (cGnRH2) and zebrafish GnIH-3 (zGnIH-3) on the male gonadotropin and GnRH system expression using pituitary explants and brain slices from a neotropical species with economical and ecological relevance, Astyanax altiparanae. Our results showed that in males, cGnRH2 increased fshb and lhb mRNA levels in the pituitary explants. Interestingly, zGnIH-3 has no effect on basal gonadotropin expression, however zGnIH-3 decreased the cGnRH2-induced fshb and lhb transcripts in male pituitary explants. In the male brain slices, zGnIH-3 showed stimulatory effects, increasing gnrh2 mRNA levels. Overall, our results suggested that GnIH seems to have dual regulatory actions on gonadotropin and GnRH2 expression of A. altiparanae males. This study provided basic information on endocrine regulation of A. altiparanae reproduction, and the obtained results will expand our knowledge, improving the reproductive management of this economically important freshwater species.
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http://dx.doi.org/10.1016/j.ygcen.2018.08.006DOI Listing
March 2019

Ovarian development and the reproductive profile of Astyanax altiparanae (Teleostei, Characidae) over one year: Applications in fish farming.

Theriogenology 2017 Aug 28;98:1-15. Epub 2017 Apr 28.

Fish Endocrinology Laboratory, Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of Sao Paulo, Av. Prof Lineu Prestes n. 1524, lab 426, CEP 05508-000, Sao Paulo, SP, Brazil.

Astyanax altiparanae is a Brazilian species of substantial commercial, environmental and scientific importance; however, existing studies on its reproduction do not seem to provide enough details. In light of the increasing use of this species in fish farming and the need for basic studies for the development of new production technologies, we describe the structural and ultrastructural characteristics of the ovaries of A. altiparanae, and characterize the species' reproductive cycle. Females were collected monthly from March 2013 to February 2014, and reproductive management began in October 2013. The ovaries were removed, fixed in Karnovsky's fixative, and prepared for light microscopy, transmission electron microscopy and immunohistochemistry anti-PCNA. These techniques enabled us to characterize the ovaries, the germ cells, and the somatic cells in detail, as well as their changes over time. The reproductive cycle was characterized based on the monthly variation in the gonadosomatic ratio, the proportion of germ cells, and the rate of oogonium proliferation. The macroscopic analysis of the ovaries suggests that the vascularization pattern and color of the ovaries vary according to development. There are new types of analyses that can be applied even in the fish farming industry, such as a comparison between ovaries staining and weight or the frequency distribution of these colors throughout the year. This study also provides details on microscopic characteristics that have never before been reported for species of Astyanax, such as the presence of annulate lamellae in oogonia, the development of the zona pellucida from oocytes in the one-nucleolus step, and the development of the micropylar apparatus in oocytes in the cortical alveolar step. When the reproductive cycle was analyzed, this species was found to have a long period of spawning, with a reproductive peak from October to February and multiple spawning events, confirming the period already described in the literature. Variations in reproductive periods and the ability to reproduce in lentic environments suggest that A. altiparanae has the ability to respond quickly to environmental changes and exhibits high reproductive flexibility. All of these characteristics confirm the great potential of this species in the fish farming industry.
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http://dx.doi.org/10.1016/j.theriogenology.2017.04.044DOI Listing
August 2017