Maria João Rocha

University of Porto | UP · Department of Microscopy

Neurotransmitters modulate gonadal maturation in bivalves. However, it remains unclear whether there are differences in the nervous system structure between sexes, maturation, and ganglia. Therefore, a stereological study was conducted on the ganglia of adult peppery furrow shell (Scrobicularia plana). Equal-sized males, females, and undifferentiat...

The concentrations of seventeen endocrine disruptor compounds (EDCs) that included oestrogens, phytoestrogens, sitosterol, and banned industrial pollutants were investigated at ten sites of the Douro River estuary. Surface waters were collected during 2019. After evaluating the physicochemical data (ammonia, nitrates, nitrites and phosphates), the...

Synthetic progestins (PGs) are a large family of hormones used in continuously growing amounts in human and animal contraception and medicinal therapies. Because wastewater treatment plants (WWTPs) are unable to eradicate PGs after excretion, they are discharged into aquatic systems, where they can also be regenerated from conjugated PG metabolites...

Anthropogenic activities can introduce polycyclic aromatic hydrocarbons (PAHs) in coastal waters. Here, 16 priority PAHs were quantified by gas chromatography-mass spectroscopy (GC–MS) in seawater samples from three Iberian Atlantic habitats. Results showed global concentrations of ≅9 ng/L in the aqueous phase (DAP) and ≅94 ng/g, dw in suspended pa...

Herein, sixteen priority sixteen priority polycyclic aromatic hydrocarbons (PAHs) and seven polychlorinated biphenyls (PCBs), considered seawater quality indicators, were quantified in four beaches of the Iberian (Portuguese) northwest coastline, including one ornithological reserve. These seashores are close to an important industrial region and a...

The main aim of this study was to histochemically estimate the relative copper content in the liver during the breeding cycle of female Ohrid trout, a wild population endemic to the Ohrid Lake. The histology of the oogenetic cycle in this species was documented for the first time. Histochemistry with semi-quantification revealed positive correlatio...

The goldfish is a model organism showing great potential for research, particularly in comparative endocrinology concerning the neuroendocrine signalling and regulation of vertebrate reproduction. Furthermore, this teleost is increasingly stressed as a relevant alternative to more common fish model organisms, namely zebrafish. However, quality desc...

The liver hosts numerous vital functions, such as biotransformation and excretion of xenobiotics. Synthetic oestrogens influence liver structure and function, leading to adaptations or to dysfunctions/injury. They are often stated to induce increases in fish liver weight, but there is controversy regarding how: if by changes in hepatocyte size (hyp...

Estrone, 17β-estradiol, 17α-ethynylestradiol, alkylphenols and alkylphenol ethoxylates, are endocrine disruptor compounds. In Portugal, the report of male fish feminization in Minho, Ave and Mondego River estuaries made pertinent their evaluation in surface water samples. Data, got by specific enzyme-linked immunosorbent assays, showed that estroge...

Iron is an essential micronutrient for the normal metabolism of all cells. Intracellular iron varies within normality ranges, and depletion or overload of brings serious consequences. As such, iron can also be deleterious when appearing as an environmental toxicant. The aim of this study was to estimate semi-quantitatively the liver iron content in...

Pharmaceuticals contamination (e.g., synthetic progestins), and global climate change, represent two of the most stressful factors affecting aquatic species. To our knowledge, there is huge gap of data regarding the combined effects of both stressors on vertebrates’ reproduction. Thus, it is crucial to implement rapid screenings of measurable histo...

A novel histological and ultrastructural study of the three ganglia types of Scrobicularia plana was made. Adults of both sexes and immature specimens were collected at Ria Formosa lagoon, Portugal. For light microscopy, animals fixed in formalin were processed for various staining, including serotonin and dopamine immunohistochemistry. For transmi...

As a case study, the estuary of the international Douro River (Iberian Peninsula) was sampled over a year (2010) at six sampling sites to determine the presence of 56 pesticides of different categories (insecticides, herbicides, and fungicides). 96% of measured pesticides were detected in 79% of the quantified samples. Individual average pesticide...

Given the lack of knowledge regarding climate change-chemical exposure interactions, it is vital to evaluate how these two drivers jointly impact aquatic species. Thus, for the first time, we aimed at investigating the combined effects of increased temperature, pCO2 and the synthetic progestin levonorgestrel on survival, growth, consumption rate an...

Interactive effects between multiple stressors, namely climate drivers (e.g., temperature) and chemical pollution (e.g., endocrine disruptors) are poorly studied. Here, it was for the first time evaluated the combinatory effects of temperature and a synthetic progestin, levonorgestrel (LNG), on the fitness and reproductive-related endpoints of zebr...

The nervous system of bivalves is bilaterally symmetrical and consists of interconnected cerebropleural, pedal and visceral ganglia, which may be partially to totally fused. We studied the microanatomy of the ganglia of Scrobicularia plana using three-dimensional (3D) reconstruction. We also examined whether intersex differences in the neural struc...

This study investigated the presence of 16 priority polycyclic aromatic compounds (PAHs) in waters from the Douro River estuary and nearby Atlantic seacoast, which both bath the Porto metropolis. In the area, there is an oil refinery, an important harbour, an intense maritime traffic, small marinas and highly inhabited cities. For the analysis of P...

This study on wild female Ohrid trout was primarily designed to provide a general overview of the breeding cycle influence upon selected aspects of hepatocytes. According with a semiquantitatively evaluation, some of these cell's structural compartments change during the breeding cycle. Structural modifications were disclosed in the relative occurr...

A method for quantitative determination of 55 pesticides in a bivalve matrix was established, based on QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction and using gas chromatography (GC)-ion trap (IT) mass spectrometry (MS/MS). Accomplishing the European SANCO guidelines, this method was validated using 5 g of homogenized soft t...

The Mira River is a Portuguese water body widely known for its wilderness and is advertised as one of the less polluted European rivers. On this presumption, the levels of endocrine-disrupting compounds (EDCs) in Mira waters were never measured. However, because environmentalists have claimed that the Mira could be moderately polluted, a range of 1...

The origin and diversification of the metazoan endocrine systems represents a fundamental research issue in biology. Nuclear receptors are critical components of these systems. A particular group named VDR/PXR/CAR (NR1I/J) is central in the mediation of detoxification responses. While orthologues have been thoroughly characterized in vertebrates, a...

Ria de Aveiro is a Portuguese lagoon renowned for its ecological and economic importance. Nonetheless, in literature, few data exist about its organic pollution. Accordingly, this study chemically monitored for the first time a series of 17 endocrine disruptor compounds (EDCs). The target EDCs include natural (17β-oestradiol, oestrone) and pharmace...

This study describes the simultaneous quantification of 56 pesticides in surface coastal water, supported by the development and validation of a gas chromatography (GC)–ion trap (IT) mass spectrometry (MS) method. Samples (500 mL) were pre-concentrated 2500 times by solid phase extraction (OASISTMHLB). The compounds were identified and quantified,...

Two novel methods were developed to extract and quantify 56 pesticides in surface waters, considering their content in both dissolved aqueous phase (DAP) and suspended particulate matter (SPM) fractions. These procedures were applied to coastal samples taken seasonally during 2012-2013, from three strategic sampling sites along the Ria Formosa Lago...

Oogenesis in zebrafish (Danio rerio) is controlled by the hypothalamus- pituitary gonadal axis and reproductive hormones. In addition, an interference of stress hormones is known with reproductive biology. In the presented work, we aimed to explore the hypothesis that cortisol (Cort) and adrenocorticotropic hormone (ACTH) may affect early oogenesis...

The Tagus is the longest Iberian river, notwithstanding, the levels of natural and xenoestrogenic endocrine-disrupting compounds (EDCs) were never measured in its estuary. Suspecting for their presence, we made a major survey of 17 EDCs that include: (i) natural (17β-oestradiol and estrone) and pharmaceutical oestrogens (17α-ethynylestradiol); (ii)...

A Stereological Estimation of the Nervous Ganglia Volumes and Number of Neurons in the Peppery Furrow Shell Scrobicularia plana (da Costa, 1778) - Volume 21 Issue S5 - Sukanlaya Tantiwisawaruji, Ana Silva, Uthaiwan Kovitvadhi, Miguel A. Pardal, Maria J. Rocha, Eduardo Rocha

Kinetics of the Metabolic and Morphological Alterations in Brown Trout Hepatic Peroxisomes Under Estradiol Influence - Volume 21 Issue S5 - Carla Batista-Pinto, Alexandre Lobo-da-Cunha, Fernanda Malhão, Albina D. Resende, Maria J. Rocha, Eduardo Rocha

Ethinylestradiol Exposure of Primary Culture Brown Trout Hepatocytes Induce Morphological Changes in Peroxisomes - Volume 21 Issue S5 - Ivone Pinheiro, Fernanda Malhão, Tânia V. Madureira, Maria João Rocha, Rogério A.F. Monteiro, Eduardo Rocha

Stereology of Brown Trout Liver Peroxisomes at Vitellogenesis and Pre-spawning Strengthens the Hypothesis of Their Regulation by Sex Steroids - Volume 21 Issue S5 - Eduardo Rocha, Albina D. Resende, Maria J. Rocha, Fernanda Malhão, Rogério A.F. Monteiro, Alexandre Lobo-da-Cunha

Here, it is exposed the environmental concentrations of sixteen oestrogenic EDCs, which were chosen taking in account the following features: (i) in vivo and in vitro potency, such as the natural oestrogens and EE2 [12]; (ii) abundance in terms of incidence and concentration), such as bisphenol A and the alkylphenols (and their ethoxylates) [13, 14...

The basic pathway of oocyte development and its regulation is evolutionarily conserved among vertebrates; however, little is known about the role of hormones at the first stages (Stages I and II) of follicle development in fish. In the present study, zebrafish follicles at Stages I and II were exposed in vitro to the reproductive hormones 17β-oestr...

The Lima River is a Spanish-Portuguese water body. Notwithstanding the fact that the river incorporates protected natural areas, levels of endocrine-disrupting compounds (EDCs) within its waters have never been measured; such EDCs include the following: natural and pharmaceutical oestrogens (17β-estradiol, E1, and 17α-ethynylestradiol), industrial...

The Douro River lower basin water quality was studied regarding its concentration on 18 trace elements (Be, Al, Cr, Mn, Co, Ni, Cu, Zn, As, Se, Mo, Ag, Cd, Sb, Ba, Tl, Pb and U, measured by ICP-MS). Other physicochemical parameters, such as pH, conductivity, dissolved oxygen and water temperature were also determined in situ. To take into account t...

A new and fully validated QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction and gas chromatography-tandem mass spectrometry methodology was developed and subsequently implemented for the quantification of 16 polycyclic aromatic hydrocarbons (PAHs) in wild (from Matosinhos Beach, Portugal) and commercial (from Ria de Arousa, Spai...

Fish are bioindicators of water pollution, and an increased rate of their erythrocyte nuclear morphological abnormalities (ENMAs)-and particularly of erythrocyte micronuclei (EMN)-is used as a genotoxicity biomarker. Despite the potential value of ENMAs and MN, there is scarce information about fish captured in Iberian estuaries. This is the case o...

The Mondego River estuary demonstrates signs of pollution, but the levels of endocrine disrupting compounds (EDCs), such as the natural (17β-estradiol and estrone) and pharmaceutical (17α-ethynylestradiol) estrogens, xenoestrogenic industrial pollutants (4-octylphenol, 4-nonylphenol, and their mono- and diethoxylates and bisphenol A), phytoestrogen...

Liver lesions in wild fish have been associated with xenobiotic exposure. Facing reports of pollution in the Douro River estuary (north of Portugal), we have been making field surveys using fishes and targeting histopathological biomarkers of exposure and effect. Herein, we intended to better characterize and report the rate of one poorly understoo...

Abstract To achieve more information about growth and development of oocytes in teleost fish or concerning toxicity testing, it is necessary to develop adequate in vitro oocyte culture conditions. Herein, initial stages of zebrafish oocytes (I, primary; II, cortical; III, vitellogenic) were analysed under serum-free medium conditions as gonadal fra...

A newly analytical method based on QuEChERS extraction followed by gas chromatography with mass spectrometry (GC-MS) analysis was developed and validated for the quantification of 18 PCBs in wild (from Matosinhos Beach, Portugal) and cultivated (from Ria de Arousa, Spain) mussel samples, pooled by sex. Wild animals showed higher PCB levels than cul...

A gas chromatography with mass spectrometric detection (GC-MS) method was developed and optimized for the determination of 17 endocrine disrupting compounds (EDCs) in coastal water samples. The evaluated EDCs were from different origins and included estrogens, bisphenol A, alkylphenolethoxylates, alkylphenols, phytoestrogens and sitosterol (SITO)....

The purpose of this study was to evaluate biochemical and morphological responses induced by carbaryl in the liver of Nile tilapia (Oreochromis niloticus) exposed during 21 days to sublethal concentrations (0.25 and 0.5 mg L−1), testing also recover for 14 days in clean water, after 14 days exposure. The activities of the following enzymes were mea...

In spite of its outstanding ecological and touristic importance the Ria Formosa Lagoon shows signs of anthropogenic pollution. Nonetheless, until the present survey no studies had ever documented the measurement of natural and pharmaceutical estrogens (17β-estradiol, estrone, and 17α-ethynylestradiol), xenoestrogenic industrial pollutants (4-octylp...

The Ave River flows through the most urbanized and industrialized Porto district areas. Despite ovotestis was recently reported in male fish from the estuary, no data exist on the water levels of natural and pharmaceutical estrogens (17β-estradiol, estrone, and 17α-ethynylestradiol), xenoestrogenic industrial pollutants (4-octylphenol, 4-nonylpheno...

The Sado waters show signs of anthropogenic pollution. However, hardly any information exists on the levels of endocrine disrupters (EDCs) such as estrogens (17β-estradiol, estrone and 17α-ethynylestradiol) and there is no information at all on xenoestrogenic industrial pollutants (4-octylphenol, 4-nonylphenol and their mono and diethoxylates and b...

A negative correlation between female gonadal maturation kinetics and size variations of hepatic peroxisomes was earlier documented in brown trout, as a probable impact of serum estrogen changes during the reproductive cycle. Herein, we investigated whether the organelle volume/surface dynamics seen in female brown trout liver peroxisomes - without...

In natural environments fish populations are exposed to many potential xenoestrogens, whereby understanding the impacts of mixtures continue to be of great interest. The main objective of this study was, therefore, to understand whether and how an environmentally relevant mixture of xenoestrogens found in the Douro River estuary can disrupt the nor...

Qualitative and quantitative approaches were tested to assess zebrafish liver effects after sub-acute exposures of certain pharmaceuticals. Carbamazepine, fenofibric acid, propranolol, sulfamethoxazole and trimethoprim were tested individually and in mixtures, including low environmental levels. Overall, data showed sex specific reactions in liver,...

In natural environments fish are exposed to endocrine disrupting compounds (EDCs) present at low concentrations and with different modes of actions. Here, adult zebrafish of both sexes were exposed for 21 days to an estrogenic mixture (Mix) of eleven EDCs previously quantified in Douro River estuary (Portugal) and to 100 ng/L 17α-ethinylestradiol (...

The Leça River and two beaches close to the north and south margins of the estuary, nowadays an important seaport harbor, show signs of contamination. However, the chemical nature of that contamination is uncertain. Therefore, this study checked for the presence of 11 endocrine disrupting compounds (EDCs) of animal (17β-estradiol and estrone), phar...

Changes in fish macrophages (Macs) are useful indicators of environmental pressures, but responses due to chemical and nonchemical stresses may be confounded by natural sources of variability. These may include sex and gonadal stage. In this study, we start addressing the following question: is the seasonally dependent ovary stage a factor to consi...

Quantitative microscopic studies of season- and breeding-related changes in the liver of farm-raised fish are very scarce, and those of wild fish populations are virtually nonexistent. Moreover, none of the available studies investigated breeding-related changes in hepatic stroma, although it is already known that changes might exist in the parench...

Fish reproduction is characterized by a puzzling diversity of strategies regarding how different species determine their sex, produce male and female fertile gametes, and mate. Maybe the success of the teleost group in conquering so many different habitats around the world are owed, at least in part, to these special features. Sex in fishes may be...

A simple low-cost, analytical method based on microwave-assisted extraction of sediments, followed by solid phase micro-extraction and gas chromatography mass spectrometry, was developed and validated for the quantification of sixteen polycyclic aromatic hydrocarbons (PAHs) in marine and estuarine sediment samples. The PAHs were those included in t...

This paper describes the development and validation of a GC-MS method which allows the simultaneous quantification of 11 endocrine disrupting compounds (EDCs) in surface water samples from both estuary and sea. The analysed EDCs are oestrone (E1), 17β-estradiol (E2), 17α-ethynylestradiol (EE2), 4-tert-octylphenol, 4-n-octylphenol, 4-nonylphenol, bi...

Fish embryos are a particularly vulnerable stage of development, so they represent optimal targets for screening toxicological effects of waterborne xenobiotics. Herein, the toxicity potential of two mixtures of pharmaceuticals was evaluated using a zebrafish embryo test. One of the mixtures corresponds to an environmentally realistic scenario and...

Many environmental pollutants can exert adverse effects on exposed organisms, including fish, leading to disruption of the endocrine system. Enzymes involved in the sex steroid biosynthesis are potential targets for the toxic action of pollutants. In this context, we investigated the hypothesis that selected estrogenic chemicals-the pharmaceutical...

Concerns associated with pharmaceuticals in aquatic systems demand the establishment of links between xenobiotics and their respective concentrations and impacts on aquatic organisms. Herein, effects of non-steroidal pharmaceuticals in the gonadal maturation of zebrafish (Danio rerio) were evaluated by histopathological and stereological analyses a...

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Previous studies in the Douro River estuary, based on occasional sampling, showed the presence of several estrogenic disrupting chemicals (EDCs). In sequence, we hypothesized that such type of pollution was more likely an enduring issue than an occasional phenomenon, and that it may even affect recreational beaches in each side of the estuarine mou...

An analytical method based on solid-phase extraction followed by gas chromatography mass spectrometry (GC-MS) was developed and validated for the quantification of 39 pesticides with distinct physico-chemical characteristics (including some degradates), in estuarine water samples. The method includes pre-concentration of samples (2500-fold factor),...

The amount and distribution of six pharmaceutical compounds belonging to distinct therapeutic classes were investigated along the navigation channel of the Douro River estuary. Distinct spatial and temporal trends were considered and a total of 87 water samples were pre-concentrated by solid-phase extraction (SPE) and analyzed by liquid chromatogra...