Molecular identification of sharks from the genus Sphyrna (Elasmobranchii: Chondrichthyes) in Maranhão Coast (Brazil).

Sharks of the genus Sphyrna are under intense exploitation globally. In Brazil's northern coast, this genus represents a high proportion of fisheries landings and comprises four species. However, due to difficulty of specific identification when specimens are landed, most of the records are limited to the genus level. Here we analyzed the effectiveness of ITS2 (Internal Transcribed Spacer 2 of rDNA) fragment length protocol (Abercrombie et al., 2005) for identifying hammerhead shark species, comparing with the analysis of COI (Cytochrome oxidase subunit I) and ITS2 sequences. We evaluated samples of muscle tissue acquired in the main fishing ports of Maranhão: Carutapera, Raposa e Tutóia. Sampling was conducted between March 2017 to March 2018 and complemented with material deposited in collection (2015). COI results indicated the occurrence of endangered species which are prohibited to be landed. These include Sphyrna mokarran (67%), S. lewini (15%), S. tudes (3%), and S. tiburo (15%). For the ITS2 marker, we investigated the optimization of the protocol developed by Abercrombie (2005) for to improve the use in this geographical area througout design of a new primers.

Gill and hepatic histological alterations in Sciades herzbergii resulting from trace element contamination in the Port of São Luiz, Brazil.

The objective of this study was to evaluate, through changes in the gills and livers of Sciades herzbergii, the environmental contamination to which estuarine organisms are exposed in two areas in São Marcos Bay - MA. Two collection areas located in São Marcos Bay were selected for this study: A1, an area close to the Port Complex, and A2, an area on Caranguejos Island (included in the Environmental Protection Area of Baixada Maranhense). Collections were carried out during rainy and dry periods. Sediments (for trace element analyses), surface water (for physico-chemical analyses), and specimens of S. herzbergii (for biometric measurements and identification of branchial and hepatic histopathology) were collected. Physico-chemical parameters (pH, dissolved oxygen, temperature and salinity) were within limits established by Brazilian legislation. Arsenic (in A1) and nickel (in A1 and A2) were above the legal standards in both periods. The highest percentage of histological alterations in the gills (aneurysms, lamellar fusion and detachment of the epithelium) occurred in the port area, in the rainy (93%) and dry (74%) periods. Liver alterations (melanomacrophage centers and necrosis) occurred only in specimens from the same area, in the rainy (41%) and dry (36%) periods. The highest histological indices of gill and liver changes were recorded in A1. This result was further supported by the total HI value of the lesions, which was higher in the port area compared to A2 (less impacted area), suggesting that the environmental conditions in that location are less favorable for the well-being of these organisms. Permanent environmental monitoring of the area is necessary to control environmental impacts efficiently.

Impacted estuaries on the Brazilian Amazon coast near port regions influence histological and enzymatic changes in Sciades herzbergii (Ariidae, Bloch, 1794).

Enzymatic (glutathione S-transferase, GST and catalase, CAT) and histological biomarkers in S. herzbergii are important for the analysis of impacted estuaries in port regions of the Brazilian Amazon coast. Fish specimens were collected in two areas in the rainy and dry seasons: Porto Grande (potentially impacted region) and Ilha dos Caranguejos (less impacted region). Sediment samples were collected for chemical analysis. Morphometric, histological, and enzymatic biomarker analyzes were performed. The analysis of the sediments collected in the potentially impacted region showed levels of iron, aluminum and polycyclic aromatic hydrocarbons above the limits allowed by CONAMA legislation. Histological changes in the gills and liver, as well as GST and CAT activities, were high in fish collected at the port. Analyzes suggest that fish in the potentially impacted region are subject to pollutants that compromise their health.

Aquatic pollutants are associated with reproductive alterations and genotoxicity in estuarine fish (Sciades herzbergii - Bloch, 1794) from the Amazon Equatorial Coast.

Estuaries are constantly subject to continuous environmental impacts of human activities, such as fisheries, port or industry, and domestic sewage, with fish being one of the most affected aquatic animals, reflecting the impacts directly on their bodies. Thus, the aim of this study was to carry out the biomonitoring of an estuary located on the Amazonian Equatorial Coast through analysis of PAHs (Polycyclic Aromatic Hydrocarbons) in the water, in addition to trace metals, histopathological alterations and analysis of erythrocyte micronuclei in Sciades herzbergii. S. herzbergii was used as a model species, due to its estuarine-resident behavior. Gonad and gill samples were subjected to histopathological evaluations. The quantification of trace metals was performed in samples of skeletal muscles of the animals collected, where concentrations of Lead (Pb), Copper (Cu), Zinc (Zn), Cadmium (Cd), Magnesium (Mg), Iron (Fe) and Aluminum (Al) were found. Except for Cadmium (Cd), all the concentrations were above the recommended limits. The PAHs analysis revealed the presence of Naphthalene and Acenaphthene in the water samples Histopathological and genotoxic analyses revealed of lesions in 100% of the study specimens. Thus, the histological and genotoxic alterations found in 100% of S. herzbergii specimens captured in São José Bay-MA are potentially associated with PAH concentrations present in the water. These results are potentially associated with the presence of PAH and trace metals, both in water and in animal tissues, inferring a general scenario of environmental contamination which directly implies a risk to the health and survival of the local biota. This study shows the relevance of continuous biomonitoring of estuarine ecosystems, in order to guide authorities regarding sewage management and ensure the evolutionary development of estuarine species, especially fishes of importance in the local cuisine, therefore related to human food security.

Histological biomarkers and biometric data on trahira Hoplias malabaricus (Pisces, Characiformes, Erythrinidae): a bioindicator species in the Mearim river, Brazilian Amazon.

The objective of this study was to evaluate the levels of environmental contamination in a protected area in the Brazilian Amazon. For this, two areas were chosen along the Mearim River: the reference area (A1) and the potentially contaminated area (A2), where water samples were collected, for physicochemical and microbiological analyses, as well as specimens of Hoplias malabaricus, for the evaluation of biometric data and incidence of branchial lesions. The physicochemical analyzes of the water from both areas showed contamination (low levels of dissolved oxygen, tubidity and high iron concentrations, especially in A2). The microbiological analyzes showed that all water samples showed total coliform values higher than those acceptable by CONAMA and WHO (with higher values in A2), in addition to E. coli values higher than those allowed by legislation in A2. Regarding biometric data, male and female fishes were significantly longer and heavier in A1 during the dry and rain seasons and the gonadosomatic index also showed higher values in A1 than in A2 in both seasons. H. malabaricus showed gill lesions of minimal to moderate pathological importance in A1 and A2, indicating that specimens from both areas of the Mearim River showed biological responses to contamination. The observed changes in the water quality, bimetic parameters and the histological analyzes of the specimens of H. malabaricus directly reflect on the quality and health of the fishes in the Mearim River, and point to the urgent need for prevention and remediation of contamination in these ecosystems.

Aggregation of hepatic melanomacrophage centers in S. herzbergii (Pisces, Ariidae) as indicators of environmental change and well-being / [Agregação de centros de melanomacrófagos hepáticos em S. herzbergii (Pisces, Ariidae) como indicadores de mudança ambiental e de bem-estar]

The melanomacrophage centers (MMCs) in the liver of fish are indicators of environmental conditions, as they are involved in xenobiotic biotransformation. The objective of this study was to evaluate the number of MMC in the liver of juveniles and adults of Sciades herzbergii from areas with different levels of contamination. The fish were caught at three points (reference - A1, potentially impacted - A2 and contaminated - A3), in São José bay (Maranhão, Brazil), in four samples. The livers were subjected to the standard histological procedure and 5µm sections were stained with hematoxylin-eosin. In livers of A2 adult individuals (260.50±161.50 MMCs / mm²) they presented a greater number of MMCs when compared to A3 adults (60.00 ± 30.10 MMCs / mm²). Juveniles showed considerable values in A1 (100.00 ± 0.00 MMCs/mm²) and A2 (95.33 ± 33.00 MMCs / mm²) compared to juveniles in A3 (49.00±0.00 MMCs/mm²). These high values are unexpected for young people. The average number of MMC correlated with the rainy season in the region. The use of hepatic MMCs as a biomarker of exposure to pollutants, in particular substances from fisheries systems, such as ammonia and nitrite, proved to be adequate to differentiate areas with different levels of impacts.(AU)

Os centros melanomacrófagos (MMCs) no fígado de peixes são indicadores das condições ambientais, pois estão envolvidos na biotransformação xenobiótica. O objetivo deste estudo foi avaliar o número de MMC no fígado de juvenis e adultos de Sciades herzbergii de áreas com diferentes níveis de contaminação. Os peixes foram capturados em três pontos (referência - A1; potencialmente impactado - A2; e contaminado - A3), na baía de São José (Maranhão, Brasil), em quatro amostras. Os fígados foram submetidos ao procedimento histológico padrão e cortes de 5µm foram corados com hematoxilina-eosina. Em fígados de indivíduos adultos A2 (260,50±161,50 MMCs/mm²), eles apresentaram maior número de MMCs quando comparados aos adultos A3 (60,00±30,10 MMCs/mm²). Os juvenis apresentaram valores elevados em A1 (100,00 ± 0,00 MMCs/mm²) e A2 (95,33±33,00 MMCs/mm²) quando comparados aos juvenis em A3 (49,00±0,00 MMCs/mm²). Esses altos valores são inesperados para os jovens. O número médio de MMC correlacionou-se com a época chuvosa na região. A utilização de MMCs hepáticos como biomarcador de exposição a poluentes, em particular substâncias provenientes de sistemas pesqueiros, como amônia e nitrito, mostrou-se adequada para diferenciar áreas com diferentes níveis de impactos.(AU)

Biomarcadores enzimáticos e histológicos em brânquias de Ucides cordatus (Linnaeus, 1763) (Crustacea, Brachyura, Ucididae) indicativos de impactos ambientais em uma região portuária do nordeste do Brasil / [Enzymatic and histological biomarkers in Ucides cordatus (Linnaeus, 1763) (Crustacea, Brachyura, Ucididae) gills indicative of environmental impacts in a port region of northeastern Brazil]

Objetivou-se neste estudo analisar biomarcadores histológicos e bioquímicos em brânquias de U. cordatus indicativos de impactos na Baía de São Marcos. Caranguejos foram coletados em quatro áreas na Baía de São Marcos: A1= Ilha dos Caranguejos (com baixo impacto); A2= Coqueiro, A3= Porto Grande, A4= Cajueiro (áreas potencialmente impactadas). Mediram-se os dados biométricos de cada exemplar de caranguejo. Amostras de brânquias foram submetidas à técnica histológica padrão e homogeneizadas em tampão fosfato, e o sobrenadante foi utilizado para análise das enzimas glutationa-S-transferase (GST) e catalase (CAT). A biometria indicou que os caranguejos de A1 são significativamente (P<0,05) maiores e mais pesados do que os caranguejos das áreas A2, A3 e A4. As alterações branquiais (rompimento das células pilastras, deformação do canal marginal, deslocamento da cutícula e necrose) foram significativamente (P˂0,05) mais frequentes em caranguejos de A2, A3 e A4 do que nos caranguejos de A1. As atividades enzimáticas da GST e CAT nos caranguejos apresentaram diferença significativa (P<0,05) entre as áreas de coletas, com padrão similar ao observado para as alterações branquiais. Os biomarcadores analisados mostraram que os caranguejos estão sob diferentes níveis de impactos (A4>A3>A2>A1) ao longo da Baía de São Marcos.(AU)

The objective of this study was to analyze histological and biochemical biomarkers in U. cordatus gills indicative of impacts in São Marcos Bay. Crabs were collected from four areas in São Marcos Bay: A1=Ilha dos Caranguejos (with low impact); A2=Coqueiro, A3=Porto Grande, A4=Cajueiro (potentially impacted areas). The biometric data of each specimen was measured. Gill samples were submitted to standard histological technique and homogenized in phosphate buffer, and the supernatant was used for analysis of glutathione S-transferase (GST) and catalase (CAT) enzyme activity. Biometric data indicated that crabs found in A1 are significantly (P<0.05) larger and heavier than crabs found in A2, A3 and A4 areas. Gill alterations (rupture of pilaster cells, Dilation of the marginal channel, Cuticle Rupture and necrosis) were significantly (P˂0.05) more frequent in the crabs in A2, A3 and A4 than crabs in A1. The enzymatic activities of GST and CAT showed significant difference (P<0.05) between the sampling areas, similar to that observed for gill alterations. The biomarkers analyzed showed that the crabs are under different impact levels (A4> A3> A2> A1) along the São Marcos Bay.(AU)

Biomarcadores histológicos em Sciades herzbergii (Pisces, Ariidae) para avaliação de impactos em ambientes estuarinos da Baía de São Marcos, Maranhão / [Biomarkers histological in Sciades herzbergii (Pisces, Ariidae) for impact assessment in estuarine environments of São Marcos Bay, Maranhão]

Neste estudo, objetivou-se avaliar o impacto ambiental em áreas estuarinas da Baía de São Marcos, Maranhão, utilizando-se alterações histológicas nas brânquias e no fígado de Sciades herzbergii como biomarcadores de contaminação aquática. Foram realizadas coletas no período chuvoso (março de 2018) e de estiagem (julho de 2018), em que foram capturados 40 exemplares de S. herzbergii em duas regiões distintas da Baía de São Marcos: A1) Ilha dos Caranguejos (área referência); A2) Terminal de Pesca Industrial de Porto Grande (área potencialmente contaminada). As estações foram georreferenciadas por GPS e, em cada local de captura dos peixes, registraram-se as seguintes variáveis abióticas: temperatura, salinidade, pH e oxigênio dissolvido. Em campo, os organismos foram pesados e medidos, as brânquias e os fígados foram retirados e fixados. Posteriormente, foram mantidos em álcool a 70% até o procedimento histológico padrão. Resultados indicam que as duas áreas são semelhantes em termos de características físico-químicas da água e do sedimento. Todavia, a análise semiquantitativa das alterações histológicas observadas revelou que as brânquias foram o órgão mais afetado nos peixes de A2, indicando que os peixes desse local apresentam danos teciduais maiores resultantes de estresse acentuado por impactos antrópicos.(AU)

This study aimed to evaluate the environmental impact in estuarine areas of São Marcos Bay, Maranhão, using histological changes in the gills and liver of Sciades herzbergii as biomarkers of aquatic contamination. Samples were collected during the rainy (March 2018) and dry (July 2018) seasons, in which 40 specimens of S. herzbergii were captured in two distinct regions of São Marcos Bay: A1) Crab Island (reference area); A2) Porto Grande Industrial Fishing Terminal (potentially contaminated area). The stations were georeferenced by GPS and at each fish capture site the following abiotic variables were recorded: temperature, salinity, pH and dissolved oxygen. In the field, the organisms were weighed and measured and the gills and livers removed and fixed. Afterwards, they were kept in 70% alcohol until the standard histological procedure. Results indicate that the two areas are similar in terms of physicochemical characteristics of water and sediment. However, the semi-quantitative analysis of the observed histological alterations revealed that the gills were the most affected organ in A2 fish, indicating that the fish in this site have greater tissue damage resulting from stress accentuated by anthropic impacts.(AU)

Biomarcadores histológicos em brânquias de Sciades herzbergii (Siluriformes, Ariidae) capturados no Complexo Estuarino de São Marcos, Maranhão / Histological biomarkers in gills of Sciades herzbergii (Siluriformes, Ariidae) caught in the Estuarine Complex of São Marcos, Maranhão

Neste estudo, objetivou-se avaliar a qualidade ambiental de duas áreas do complexo estuarino de São Marcos, utilizando-se as alterações branquiais em Sciades herzbergii como biomarcadores de contaminação aquática. Foram realizadas coletas no período de estiagem (setembro/dezembro de 2015) e no período chuvoso (março/junho de 2016). Foram capturados 40 exemplares de S. herzbergii em dois locais distintos do complexo estuarino: A1) Porto do Itaqui Bacanga (área potencialmente impactada); A2) Igarapé do Puca, Alcântara (área controle). Foram registradas as variáveis abióticas: salinidade, temperatura, pH e oxigênio dissolvido. No laboratório, as brânquias de cada espécime foram fixadas em formol a 10% e mantidas em álcool a 70% até a execução da técnica histológica usual. As alterações branquiais identificadas nos peixes da área potencialmente contaminada foram: deslocamento do epitélio (DE), congestão (CO), aneurisma (AN), fusão lamelar (FL), hiperplasia (HI), dilatação capilar (DC), necrose (NE) e proliferação de células epiteliais (PCE). As alterações na região controle foram: deslocamento do epitélio (DE), fusão lamelar (FL), necrose (NE) e proliferação de células epiteliais (PCE). As alterações branquiais foram mais frequentes na região portuária (84%) do que na região controle (16%). Esses resultados indicam que os peixes da área potencialmente contaminada estão apresentando respostas biológicas ao estresse local.(AU)

This study aimed to compare an environmental quality of two areas of the estuarine complex of São Marcos using gills of changes in Sciades herzbergii as biomarkers of aquatic contamination. Samples were made during the dry season (September/ December 2015) and rainy season (March/ June 2016). They were captured 40 specimens of S. herzbergii at two different points of the estuarine complex: A1) Itaqui port (potentially contaminated area); A2) Igarapé do Puca, Alcântara (control area). Abiotic variables were recorded: salinity, temperature, pH and dissolved oxygen. In the laboratory, the gills of each species were fixed in 10% formaldehyde and kept in 70% alcohol until the usual histological technique was performed. The gill alterations identified in the fish of the potentially contaminated area were: epithelial displacement (DE), congestion (CO), aneurysm (AN), lamellar fusion (FL), hyperplasia (HI), capillary dilation (DC), necrosis and epithelial cell proliferation (PCE). The changes in the control region were: epithelial displacement (DE), lamellar fusion (FL), necrosis (NE) and epithelial cell proliferation (PCE). Gill alterations were more frequent in the port region (84%) than in the control region (16%). These results indicate that fish from the potentially contaminated area are presenting biological responses to local stress.(AU)

Integrated assessment of biomarker responses and microbiological analysis of oysters from São Luís Island, Brazil.

This study was conducted to evaluate the use of biochemical biomarkers and microbiological analysis to identify levels of oyster contamination at different ports in São Luís Island (Maranhão), Brazil. Oysters were analyzed for total coliforms, thermotolerant coliforms, Escherichia coli and Aeromonas spp. In addition, tissue was removed from the digestive gland to determine the glutathione-S-transferase (GST) and catalase (CAT) activity. The highest percentage of microbiological contamination of oyster samples occurred during the rainy season. The activity of GST and catalase in oysters was also higher in the rainy season, coinciding with the greatest abundance of total and thermotolerant coliforms. Among the prospective biomarkers, GST showed the best results for identification of areas with higher levels of contamination.

Biomarcadores histopatológicos na espécie Hoplias malabaricus (Pisces, Osteichthyes, Erythrinidae) em uma Unidade de Conservação de São Luís (MA) / Histopathological biomarkers in Hoplias malabaricus (Pisces, Osteichthyes, Erythrinidae) species in a protected area of São Luís (MA)

Neste estudo objetivou-se validar dois órgãos (brânquia e fígado) de Hoplias malabaricus para análise de biomarcadores de contaminação aquática na Área de Proteção Ambiental (APA) do Maracanã em São Luís-MA. Exemplares de traíra foram capturados em uma lagoa de policultivo e no Rio Ambude, localizados na APA do Maracanã. A biometria foi realizada em laboratório. Brânquias e o fígado de cada espécime foram fixados em formol a 10% e mantidos em álcool a 70% até a execução da técnica histológica usual. Cortes de aproximadamente 5µm de espessura foram corados em hematoxilina e eosina (HE). Os resultados referentes à biometria dos peixes capturados durante o período de estiagem indicaram que o comprimento total e padrão dos exemplares foram maiores quando comparados com os indivíduos do período chuvoso. As alterações hepáticas identificadas foram: lipidose, necrose, infiltração leucocitária, vacuolização do citoplasma, hemossiderina e centro de melanomacrófagos. Constataram-se as seguintes lesões branquiais: fusão lamelar, aneurisma lamelar, deslocamento e necrose do epitélio, proliferações de células do muco e dilatação capilar. As respostas biológicas de H. malabaricus, registradas em nível branquial e hepático, podem ser caracterizadas como biomarcadores de contaminação aquática. As brânquias mostraram-se como os melhores órgãos para esse tipo de análise na espécie. Esses resultados indicam que os peixes da região apresentam a saúde comprometida por poluentes presentes nos ambientes analisados...

The aim of the present work was to validate two organs (liver and gill) of Hoplias malabaricus for analysis of aquatic contamination biomarkers in the Maracanã Environmental Protected Area (APA) in São Luís-MA. Trahira animals were captured in a pond polyculture and Ambude River located in Maracanã APA. Biometric analysis was performed in the laboratory. Gills and liver of each specimen were fixed in 10% formalin and kept in 70% alcohol to running the usual histological technique. Cuts of approximately 5μm thick were stained with hematoxylin and eosin (HE). The results concerning the biometrics of the fish caught during the dry season indicated that the total and standard length were higher when compared to the rainy season. The liver changes identified were: lipidosis, necrosis, leukocyte infiltration, vacuolization of the cytoplasm, hemosiderin and melanomacrophages centres. We found the following gill lesions: melting lamellar, lamellar aneurysm displacement, necrosis of the epithelium, proliferation of mucous cells and capillary dilation. The biological responses of H. malabaricus, recorded in gill and liver level, can be characterized as biomarkers of aquatic contamination. The gills appeared as the best organs for this kind of analysis in the species. These results indicate that the fish have health problems caused by pollutants present in the analyzed environments...

Biomarcadores histológicos em duas espécies de bagres estuarinos da Costa Maranhense, Brasil / Histology biomarkers in two estuarine catfish species from the Maranhense Coast, Brazil

Neste estudo objetivou-se validar lesões branquiais como biomarcadores em peixes de importância econômica (Sciades herzbergii e Bagre bagre) capturados pela pesca artesanal em dois locais diferenciados da Baía de São Marcos, MA, a fim de selecionar um táxon como bioindicador capaz de ser utilizado em programas de biomonitoramento. O primeiro ponto (S1) foi utilizado como uma área de referência - Ilha dos Caranguejos -, e o segundo ponto (S2) foi considerado potencialmente impactado - Complexo Portuário de São Luís, MA. Encontraram-se várias alterações histológicas para as duas espécies em S2, destacando-se: estreitamento lamelar, teleangectasia, fusão e descolamento do epitélio da lamela secundária. Lesões branquiais em B. bagre coletados em S1 também foram observadas, não sendo possível diferenciar os indivíduos das duas áreas analisadas. Os dados indicam que S. herzbergii é um bioindicador mais apropriado para análise de biomarcadores de contaminação aquática.

This study aimed to validate branchial lesions as a biomarker in fish (Sciades herzbergii and Bagre bagre) caught by traditional fishing in two different places at São Marcos Bay, MA, in order to select a bioindicator taxon which can be used in biomonitoring programs. The first point (S1) was used as a reference area - Caranguejos (Crab) Island and the second point (S2) was considered potentially impacted - Port Complex of São Luis, MA. Several histological changes were found for both species in S2, including: narrowing lamellar, teleangectasy, fusion and separation of secondary lamellar epithelium. Gill lesions in B. bagre collected were also observed in S1, it is not possible to differentiate between individuals of the two areas analyzed. The data indicate that S. herzbergii is more appropriate as a bioindicator for analysis of biomarkers of aquatic contamination.

Hábito alimentar do bagre sciades herzbergii (Siluriformes, Ariidae) da ilha dos caranguejos, Maranhão, Brasil / Feeding habits of the catfish sciades herzbergii (Siluriformes, Ariidae) in caranguejos island, Maranhão, Brazil

The article describes the feeding habits of the catfish Sciades herzbergii caught in the Crabs Island (Ilha dos Caranguejos), an important coastal area of the Maranhão state, Brazil. A total of 167 individuals were collected: 110 females and 57 males. All the stomach contents were identified to the closest taxonomic level. The frequency of occurrence, biomass, numerical frequency and index of relative importance (IRI) of food items were determined for both genders. Stomach repletion was predominant, demonstrating good feeding conditions for S. herzbergii. The main alimentary items in the diet of this catfish were crustaceans, fish, oligochaets and polychaets, and the decapods crustaceans were the most representative.