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Gold mining is the largest source of mercury (Hg) pollution worldwide. The discharge of mercury in the environment bears direct human health risks and is likely to increase cascading effects throughout local food chains. In the Peruvian Amazon the mining process consists of slashing and burning trees, followed by extraction of gold-bearing sediment, amalgamation with Hg and gold recovery, leading each year to the degradation of 6,000-10,000 ha and the release of 180 metric tons of Hg per year to the enviroment. The purpose of this study was to determine soil Hg levels in soils of abandoned alluvial gold mine spoils and undisturbed forest in the Madre de Dios region, the epicenter of alluvial gold mining in Peru. We selected gold mine spoils of the two most important technologies locally applied for gold extraction, i.e., Minimally Mechanized Mining (MMM) and Highly Mechanized Mining (HMM), in the native communities of Laberinto and Kotzimba, respectively. We collected 127 and 35 soil samples (0-20cm depth) from potentially contaminated sites and undisturbed forest, respectively. Physicochemical analysis and determination of Hg levels were determined for all soil samples. None of the samples had Hg concentrations above Peruvian, Canadian and British Environmental Quality Standards for Agricultural Soil (6.6mg/kg). Hg levels in MMM and HMM were not significantly different between the two areas. The main variables explaining variation of soil Hg concentrations were the vegetation cover, soil organic matter, soil pH and clay particle content, which explained up to 80% of data set variation. Surprisingly, highest Hg concentrations were found in untouched old-growth forest bordering the mine spoils, but there was also a trend of increasing Hg concentrations with the regenerating vegetation. Our findings suggest that Hg concentrations in old mine spoils are low and shouldn't stand in the way of efforts to restore soil conditions and develop sustainable land uses. However, it is urgent to end the use of Hg in mining operation to decrease human and environmental risks., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Some heavy metals related to human activities were measured in the water of Chascomús lake. The maximum concentrations were: 0.23 μg/L for Cd, 4.28 μg/L for Cr, 22.09 μg/L for Cu, 2.49 μg/L for Ni, 3.24 μg/L for Pb and 210.76 μg/L for Zn. The values of Cd, Cr, Cr, Pb and Zn were above the Argentine National Guidelines for the Protection of the Aquatic life. The analysis of gonadal condition of pejerrey fish (Odontesthes bonariensis) from this lake did not revealed any reproductive damages. However, exposures with environmental concentrations of Cd, Cr, Cu and Zn under laboratory conditions of pejerrey males (14 days), caused a significant increase of the expression of the three variants of gnrh in the brain (within Cd exposure) and a decrease in cyp19a1b mRNA (within Cu exposure). Furthermore, at pituitary level, a decrease in fshb transcript levels was observed in the fish exposed to Cd and Cr and a decrease in the expression of both gonadotropin receptors at gonadal level in Zn exposure. Moreover, the gonads of the fish exposed to all the tested metals suffered structural damages showing shortness of the spermatic lobules, fibrosis, testis ova and the presence of piknotic cells. All these findings alert that heavy metals pollution affects the expression of key reproductive genes and gonadal structure of fish species that represent the predominant group of organisms and are considered sentinel species in the aquatic ecosystems., (Copyright © 2018 Elsevier Inc. All rights reserved.)

In the present study, we determined the hepatopancratic shbg transcript abundance and ovarian immunoreactive Shbg (ir-Shbg) localization in pejerrey females at different gonadal stages during an annual ovarian cycle. In the hepatopancreas, shbg expression remains was constant in pre-vitellogenic stages, decreased at final vitellogenesis to increase again in final maturation and atretic stages to previous levels at post-vitellogenic stages. Related to this, also we found a negative significant relation between sex steroid and shbg expression. On the other hand, in the ovary we found ir-Shbg inside of cortical alveoli, from previtellogenic stages to final maturation. This localization of Shbg in a teleost fish ovary suggests a new role for Shbg in oocytes, that may also extend the oocyte fertilization/development process., (Copyright © 2018 Elsevier Inc. All rights reserved.)

In vertebrates, the reproduction is controlled by the brain-pituitary-gonadal (BPG) axis and kisspeptin has emerged as a key player of this axis. In this study, we analyzed changes in the expression levels of kiss1, kiss2, and their receptors, kissr2 and kissr3 during gametogenesis in the BPG axis of feral Odontesthes bonariensis. In females, levels of brain kiss1 showed an increase at final maturation (Fm), while kiss2 levels were shown to be high at primary growth (Pg) stage, with no differences in the expression of their receptors. In the pituitary, kiss1 and kiss2 peaked at the cortical alveoli (Ca) stage, and kissr3 at initial vitellogenesis. In parallel, there was an increase of kiss1, kissr2 and kissr3 in the ovary during the Ca stage and both receptors again at Fm stage. In males, the four genes were highly expressed in the brain at the arrested (A) stage. In the pituitary, kiss2 peaked at spermatogonial (SG) and spermatocytary (SC) stages; while kissr3 reached a peak at the spermiogenic stage (SP). In testes, kiss1 and kiss2 significantly increased during the SG and SC stages; meanwhile, kissr2 increased at SG and SC, whereas kissr3 levels were significantly high at SC and SP stages. Taken together these results showed that the kisspeptin system in pejerrey is expressed in the three levels of the BPG axis with different expression profiles during the gonadal cycle. These findings pointed that kisspeptins have different roles in gametogenesis in this species., (Copyright © 2017. Published by Elsevier Inc.)

The endocrine mechanisms that regulate spermatogenesis and their interaction with environmental cues have been poorly studied compared with oogenesis in fish. The aim of this work was to study the spermatogenesis in pejerrey under the influence of photoperiod and water temperature fluctuation in the wild, evaluating the transcript levels of brain Gnrh variants and cyp19a1b, pituitary Gth subunits, gonadal Gth receptors, 11β-hsd, and 11-KT plasma levels. Males at spermiogenic stage were observed during spring and autumn, under a photoperiod above 11h of light and a water temperature below 23 °C. Most arrested males were observed in summer when water temperatures increased above 23 °C. Males at spermatogonial stage were mainly observed in autumn, while most males at spermatocytary stage were caught in winter. An increase of gnrh-I, cyp19a1b, fshb, gpha and 11β-hsd transcripts and 11-KT plasma levels was observed during spermatogonial and/or spermatocytary stage (early spermatogenesis). The spermiogenic stage was associated to the maximum gnrh-I gene expression level and a significant increase of Gth receptors transcripts, being this fact more evident for lhcgr. During this last gonadal stage, cyp19a1b transcript level remained high, while fshb mRNA and 11-KT plasma levels showed a significant decreased compared to that occurred at the spermatocytary stage. Also, gphα and 11β-hsd gene expression levels fell during spermiation up to similar values to those observed in arrested males. A significant correlation between 11-KT and gnrh-I, cyp19a1b, gphα, fshb, 11β-hsd transcripts, and the number of spermatocytes was observed during spermatogenesis. All these findings suggested that in pejerrey, the spermatocyte proliferation occurs mainly during winter under the stimulation of 11-KT induced by FSH through the stimulation of specific enzymes, including the 11β-hsd while spermiation occurs after photoperiod increase and with temperatures of the water below 23 °C, through the stimulation of gnrh-I, cyp19a1b and lhcgr., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The ongoing of global warming trend has led to an increase in temperature of several water bodies. Reproduction in fish, compared with other physiological processes, only occurs in a bounded temperature range; therefore, small changes in water temperature could significantly affect this process. This review provides evidence that fish reproduction may be directly affected by further global warming and that abnormal high water temperature impairs the expression of important genes throughout the brain-pituitary-gonad axis. In all fishes studied, gonads seem to be the organ more readily damaged by heat treatments through the inhibition of the gene expression and subsequent synthesis of different gonadal steroidogenic enzymes. In view of the feedback role of sex steroids upon the synthesis and release of GnRH and GtHs in fish, it is possible that the inhibition observed at brain and pituitary levels in treated fish is consequence of the sharp decrease in plasma steroids levels. Results of in vitro studies on the inhibition of pejerrey gonad aromatase expression by high temperature corroborate that ovary functions are directly disrupted by high temperature independently of the brain-pituitary axis. For the reproductive responses obtained in laboratory fish studies, it is plausible to predict changes in the timing and magnitude of reproductive activity or even the total failure of spawning season may occur in warm years, reducing annual reproductive output and affecting future populations., (Copyright © 2013 Elsevier Inc. All rights reserved.)

It has been traditionally accepted that the gonadotropins (GtHs), follicle stimulating hormone (FSH) and luteinizing hormone (LH), are synthesized and secreted only by the pituitary. However, the presence of theses hormones in extrapituitary tissues has been demonstrated in mammals, and more recently also in fish. In this study, we cloned the cDNAs and characterized the expression of FSH-β, LH-β, and glycoprotein hormone α (GPH-α) subunits from brain and gonads of male and female pejerrey Odontesthes bonariensis at different stages of gonadal maturation. In situ hybridization revealed that, in addition to their classical location in pituitary cells, the three GtH transcripts were also located in the gonads. FSH-β and GPH-α subunits were found in the cytoplasm of oogonia, previtellogenic and vitellogenic oocytes in ovaries. LH-β expression was detected in previtellogenic and vitellogenic oocytes but not in oogonia. In males, the three subunits were expressed in spermatogonia and to a lesser extent in spermatocytes. Exposure of fish to high water temperatures that impair pejerrey reproduction also induced a decrease of extrahypophyseal expression of GtH subunits., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The brain-type aromatase (CYP19A2) cDNA from pejerrey Odontesthes bonariensis was characterized. Its sequence differs from the ovarian-derived aromatase (CYP19A1) previously reported for the same species. The cDNA is 2305bp in length and the deduced protein comprises 501 amino-acids. The percentage of identity was higher when compared to other brain-derived aromatase proteins (85-63%) and lower with ovarian-derived aromatases (64-57%). Pejerrey aromatases share 61% of identity. The tissue expression analysis showed that CYP19A2 was expressed in the kidney, brain, and pituitary gland of both sexes and also in the ovary, but not in the eye, spleen, liver, gill, and testis. Semi-quantitative RT-PCR analysis of different brain areas revealed that CYP19A2 was expressed significantly higher in anterior male brain areas than in the corresponding female areas, and also when compared to posterior brain areas from both sexes. An immunological analysis using a polyclonal anti-teleost aromatase showed immunoreactive aromatase cells bordering the telencephalic ventricle and a strong signal in the ependymal cells of the preoptic area and the hypothalamus. In the optic tectum immunoreactive aromatase cells were labeled in the ventral wall and in the ependymal layer of the third and fourth ventricle with lateral projections. In the pituitary gland immunoreactive aromatase cells could be found in the rostral and proximal pars distalis. In this gland, aromatase fibers were also detected in different areas; many of them concentrated around blood vessels.

The development of gonadotropin-releasing hormone (GnRH) neurons was studied in relation to the sensitive period of thermolabile sex determination in the pejerrey Odontesthes bonariensis, an atherinid fish from South America. Fish were raised from hatching at three different temperatures: 17 degrees C (100% females), 24 degrees C (70% females), and 29 degrees C (100% males). Three groups of immunoreactive GnRH (ir-GnRH) neurons were identified at the terminal nerve ganglion (TNG), the midbrain tegmentum (MT), and the preoptic area (POA). Immunoreactive GnRH (ir-GnRH) neurons were identified in the TNG at hatching (day 0) and in the MT at day 3 at all the experimental temperatures. In the POA ir-GnRH neurons were identified in the nucleus preopticus periventricularis simultaneously with the first appearance of ir-GnRH fibers in the pituitary on days 11, 14, and 17 for larvae kept at 29, 24, and 17 degrees C, respectively. The number of ir-GnRH neurons in the TNG did not show any statistical difference between temperatures. The number of ir-GnRH neurons in the MT increased in number during the experiment for larvae kept at 17 and 24 degrees C but decreased between days 17 and 31 in larvae kept at 29 degrees C. The number of ir-GnRH neurons in the POA increased during development with a peak at day 28 for all temperatures studied and the magnitude of this peak showed a correlation with incubation temperature. These results reinforce the notion that the hypothalamus-pituitary-gonadal axis is active during sex determination in pejerrey suggesting a possible role of the central nervous system and GnRH in this process. It is also suggested that GnRH neurons located in the preoptic area might be the physiological transducers of temperature during the temperature sensitive period in this species.

The ontogeny of gonadotropin 1 (GtH1) and 2 (GtH2) cells and its possible link to gonadal sex differentiation were studied in pejerrey, Odontesthes bonariensis, by immunocytochemistry using anti-chum salmon beta-GtH1 and beta-GtH2 antisera. In adults, GtH1 cells were found in the proximalis pars distalls (PPD) close to the neurohypophysis, whereas GtH2 cells were identified surrounding GtH1 cells, at the external layer of the PPD and in the pars intermedia. Essentially the same distribution was observed in larvae. In pejerrey, the phenotypic sex is governed by the temperature during the critical period of sex determination (temperature-dependent sex determination, TSD). Female proportions vary gradually from 100% at 15-19 degrees to 0% at 29 degrees, and the critical time of TSD has been estimated to be 3-5, 2-4, and 1-4 weeks after hatching at 17, 19, and 27 degrees, respectively. Thus, the expression of both GtHs was examined weekly in larvae reared from hatching to week 11, at 17, 24, and 29 degrees. The proportion of females at 17, 24, and 29 degrees was, 100%, 78%, and 0%, respectively. Histological ovarian differentiation was noticed at 7 and 4 weeks at 17 and 24 degrees, whereas testicular differentiation occurred at 7 and 4 weeks at 24 and 29 degrees. GtH1 cells were first observed at week 3 at 29 degrees and at week 4 at 17 and 24 degrees. These cells increased in number until week 4 and then decreased, disappearing after week 6 at all temperature regimes. GtH2 cells appeared at week 2 at 24 and 29 degrees and at week 3 at 17 degrees. GtH2 cell number increased until week 3 at 29 degrees and until week 4 at 17 and 24 degrees and then temporarily decreased, thereafter increasing again. These results strongly suggest that GtH1 and GtH2 are expressed by different cells. The fact that GtH1 and GtH2 cells appear just before histological gonadal differentiation at all temperatures, together with the peak of GtH1 and GtH2 cell number during the temperature-sensitive period, suggests that GtHs are related to sex differentiation or TSD in O. bonariensis., (Copyright 2001 Academic Press.)

Changes in the pituitary cells involved in amphibian metamorphosis were studied in Bufo arenarum tadpoles with inhibited thyroid function. After hatching, larvae were treated for 5 months with potassium perchlorate (KClO4), a goitrogenic substance known to prevent absorption of iodine from water or food, resulting in impaired thyroid hormone synthesis. Treated larvae continued to grow but halted their development in premetamorphosis, showing hyperplastic thyroid glands with disorganized follicles lacking colloid. Thyrotrop (TSH), lactotrop (PRL), somatotrop (GH), and corticotrop (ACTH) cells were stained immunocytochemically and the changes observed were evaluated morphometrically using an automatic image analyzer. Pars distalis volume increased in treated larvae. Morphometric results showed that, in treated tadpoles, TSH and PRL cell populations and cell volumes increased compared to normal larvae at the same stage. Changes in the GH and ACTH cell morphometry were slight. These results indicate that in Bufo tadpoles, after chronic thyroid hormone withdrawal, TSH, PRL, GH, and ACTH cells are able to develop and that thyroid hormones exert a strong feedback control on the synthesis and storage of TSH and PRL.