Levels of Total Mercury and Health Risk Assessment of Consuming Freshwater Stingrays (Chondrichthyes: Potamotrygoninae) of the Brazilian Amazon.

Mercury is an element with potential risk to fish and those who consume it. Thus, this study aimed to determine the levels of total mercury (THg), carry out a health risk assessment related to the consumption of the freshwater stingrays Potamotrygon motoro, and determine the physical and chemical properties of the water where stingrays occur. Stingrays of the species P. motoro were obtained from the Amazon River, and samples of the animals' musculature were collected to determine THg levels. Risk assessment was conducted using pre-established formulas of estimated monthly intake (EMI), maximum monthly intake rate (IRmm), and hazard quotient (HQ). Three population scenarios were evaluated, considering both sexes and differences between rural and urban areas. There was no relationship between weight and THg concentration nor between total length and THg concentration. Higher EMI values were observed in rural children; for the IRmm, male children had the lowest consumption levels. For the hazard quotient, there was a similarity between the three age groups when comparing the male and female sexes. In addition, the representatives of the rural area always had lower values than the urban area. Freshwater stingrays, like other elasmobranchs, can be crucial animal species because they act as sentinels in studies that assess harmful chemicals like mercury.

Dietary lysine requirements of Colossoma macropomum (Cuvier, 1818) based on growth performance, hepatic and intestinal morphohistology and hematology.

This study aimed to determine the dietary lysine requirements of juvenile Colossoma macropomum tambaqui based growth performance, hepatic and intestinal morphohistology and hematology. Tambaqui were fed with levels of 6.60, 9.72, 12.84, 15.96, 19.08, and 22.20 g/kg of total lysine and those who received 15.96 g/kg dietary lysine showed higher final weight (p= 0.001) and optimized feed conversion ratio (p= 0.001). Morphohistological modifications (presence of fat droplets) were present in livers of fish fed with low levels of lysine. In the morphohistology of the intestine, the diets influenced the density of the intestinal, cell layers, height and perimeters of the villi and acid mucin secretion by the goblet cells. In the proximal intestine, mucosa layer density was greater at the level of 15.96 g/kg (p= 0.001). In the middle intestine, height (p= 0.001) and perimeter (p= 0.001) of the villi were greater at low levels of lysine (respectively, 9.72 and 12.84 g/kg dietary lysine). No differences were found for the hemoglobin, hematocrit, erythrocytes, MCV, MCH, MCHC, thrombocytes and leukocytes differentiated of fish fed with experimental diets (p>0.05). The biochemical responses were affected by the diet, both in plasma concentration (protein and cholesterol; p<0.05), and in muscle (ash and moisture; p<0.05). In conclusion, the dietary requirement for lysine for tambaqui juveniles was estimated to be 15.4 - 15.6 g/kg of diet (5.7% -5.8% dietary protein).

Morphological, cytochemical and ultrastructural aspects of blood cells in freshwater stingray species in the middle Rio Negro basin of Amazonian Brazil.

In the present work, we examined the morphology, dimensions, cytochemical staining reactions and ultrastructure of blood cells from three freshwater stingray species, Potamotrygon wallacei, Potamotrygon motoro and Paratrygon aiereba, living in the waters of the middle Rio Negro basin (Barcelos, Amazonas, Brazil). We identified erythrocytes, erythroblasts, thrombocytes and four types of leukocytes (basophils, heterophils, lymphocytes and monocytes) in the blood of these stingray species. In all the freshwater stingray species studied, the shapes and dimensions of these cells were similar to those of marine elasmobranchs. Positive PAS staining occurred in heterophils and thrombocytes, and weak staining occurred in lymphocytes and monocytes, while metachromasia only occurred in basophils. Positive Sudan Black B staining was observed in thrombocytes and lymphocytes, and weak staining occurred in heterophils. Basophils and heterophils were the only cells with positive bromophenol blue staining, while no peroxidase staining was observed in any of the four leukocyte types. This is the first study to establish the dimensions and cytochemical staining profiles of blood cells in Amazonian stingray species. Because these elasmobranch species are exported as ornamental fish to countries worldwide, this study can contribute to establishing standards for blood constituents that may be helpful in assessing the health and welfare of these fish in artificial systems.

HEMATOLOGYCAL AND BLOOD BIOCHEMISTRY PARAMETERS OF CAPTIVE BIG-HEADED AMAZON RIVER TURTLES, Peltocephalus dumerilianus (TESTUDINES: PODOCNEMIDIDAE) / Parámetros hematológicos y bioquímica sanguínea de tortugas amazónicas cautivas de cabeza grande, Peltocephalus dumerilianus (Testudinas: Podocnemididae)

ABSTRACT The determination of hematological values is used to obtain knowledge about the health conditions of animal species. The big-headed Amazon River turtles, (Peltocephalus dumerilianus) are considered one of the least known testudine species concerning their biology and health status. Herein, we determined the hematological and plasma biochemical parameters of17 (eight males and nine females) adult P. dumerilianus to provide reference interval values for clinically healthy individuals. We collected the blood samples by puncturing the femoral vein using long heparinized hypodermic syringes. Sexual dimorphism for individuals was determined by external observation of the shape of the plastron. The average values obtained for the ten hematological and biochemical parameters analyzed were red blood cell count = 0.32 million μL-1; hematocrit = 20.6 %; hemoglobin = 8.5 g dL-1; mean corpuscular volume = 681.6 fL; mean corpuscular hemoglobin = 267.8 pg; mean corpuscular hemoglobin concentration = 41.9 g dL-1; glucose = 80.6 mg dL-1, total protein = 4.1 g dL-1, triglycerides = 388.9 mg dL-1, and total cholesterol = 79.3 mg dL-1. Despite the sexual dimorphism evidenced for the species, there was no significant statistical difference between males and females for both hematological and biochemical parameters analyzed herein. Based on these results, the population is considered healthy, with parameter values coinciding with previously reported reference ranges for testudines species in the region. The results obtained in this study can be used for assessing the health status of other Amazonian turtle populations, especially in actions aimed at cultivation strategies, management, and species conservation.

RESUMEN La determinación de valores hematológicos se ha utilizado para conocer las condiciones sanitarias de algunas especies animales. La tortuga cabezona del río Amazonas, Peltocephalus dumerilianus, se considera una de las especies de testudines menos conocidas en relación a su biología y estado de salud. Aquí, determinamos los parámetros bioquímicos hematológicos y plasmáticos de 17 adultos (ocho machos y nueve hembras) de P. dumerilianus con el fin de proporcionar valores de intervalo de referencia sobre los individuos clínicamente sanos. Recolectamos las muestras de sangre perforando la vena femoral con jeringas hipodérmicas largas heparinizadas. El dimorfismo sexual de los individuos se determinó mediante la observación externa de la forma del plastrón. Los valores medios obtenidos para los diez parámetros hematológicos y bioquímicos analizados fueron: recuento de glóbulos rojos = 0,32 millones μL-1; hematocrito = 20,6 %; hemoglobina = 8,5 g dL-1; volumen corpuscular medio = 681,6 fL; hemoglobina corpuscular media = 267,8 pg; concentración media de hemoglobina corpuscular = 41,9 g dL-1; glucosa = 80,6 mg dL-1, proteína total = 4,1 g dL-1, triglicéridos = 388,9 mg dL-1 y colesterol total = 79,3 mg dL-1. A pesar del dimorfismo sexual evidente para la especie, no hubo diferencia estadística significativa entre machos y hembras para los parámetros hematológicos y bioquímicos analizados aquí. Con base en estos resultados, la población se considera saludable y los valores de los parámetros coinciden con los rangos de referencia reportados previamente de las especies de testudines en la región. Los resultados obtenidos en este estudio pueden utilizarse en la evaluación del estado de salud de otras poblaciones de tortugas amazónicas, considerando especialmente aquellas acciones dirigidas al manejo, conservación y estrategias de cultivo de la especie.

Gene Expression Changes in Epinephelus marginatus (Teleostei, Serranidae) Liver as Molecular Biomarker of Iron Ore Contamination.

The aim of this study was to determine selected molecular biomarkers of iron ore contamination in Epinephelus marginatus. Molecular biomarkers were tested by checking the relative expression of genes involved in oxidative stress, trace element regulation, and cellular damage, by RT-qPCR. Iron ore exposure caused the downregulation of ferroportin (FP) gene expression and a significant upregulation in superoxide dismutase (SOD) and cytochrome P450 1A (CYP1A) genes. Iron ore affects gene expression in E. marginatus liver, indicating that it can potentially induce toxic effects in fish. Moreover, this altered gene expression pattern may be applied in monitoring iron ore contamination in marine environments.

Growth performance, hematological responses and economic indexes of Colossoma macropomum (Cuvier, 1818) fed graded levels of glycerol.

The aim of this study was to evaluate the performance, hematological responses and economic indicators of juvenile tambaqui Colossoma macropomum fed different levels of partial replacement of corn by glycerol (0%, 25%, 50%, 75%, and 100%). The experiment was conducted for 90 days in the production of aquatic organisms lab at the Nilton Lins University, Manaus, Brazil, in a completely randomized design, and consisted of four treatments, four repetitions and two sampling times. In total, 240 juveniles were used with initial average weight and standard length of 15.32 ± 1,61 g and 8.03 ± 0.22 cm, respectively. The fish were maintained in twenty 310 L water tanks that had a closed system with no reuse, continuous aeration, siphoning and replacement of water every 48 h. Feeding was twice a day with the experimental diets, which contained 28% crude protein. The following parameters were considered: zootechnical checks (weight gain + survival %), welfare (health) (hepatosomatic index + condition factor + viscerosomatic index), economic (Economic efficiency rate + economic profitability index) and hematology (hematology + metabolites + ions). The results of the study show that diets for juvenile tambaqui can contain up to 50% replacement of corn by glycerol without compromising the fishes' development under the conditions studied.

Ichthyological ethnoknowledge of the "piabeiros" from the Amazon region, Brazil.

BACKGROUND: The capture of ornamental fish is one of the main economic activities of riverine families in the Amazon. However, studies regarding the local ecological knowledge of workers in this activity are still incipient. In view of this, we have studied and explored the local ecological knowledge of artisanal fishers who specialize in the capture of fish for the aquarium trade in the middle part of the Negro River basin and investigated issues related to the ecological aspects of the fish species that are targeted by this trade in the region. METHODS: Therefore, we conducted semi-structured interviews and applied questionnaires to artisanal fishers of ornamental fish (N = 89), from the municipality of Barcelos, from January to April 2016. RESULTS: In total, 41 popular names were cited, which correspond to four ethnocategories and 10 families. The main species were Paracheirodon axelrodi (12.5%), Hemigrammus bleheri (8.3%), Ancistrus dolichopterus (6.4%), Symphysodon discus (5.3%), and Potamotrygon motoro (3.8%). According to the fishers, the species of fish known in the region as "piabas" have a preference for living in clusters (28.9%) and carry out migratory movements (26.1%). The diet of local fish species reported by fisheries is diverse, though mainly based on periphyton (42.2%), and the reproductive cycle directly influenced by the period of flooding of rivers in the region (37.6%) CONCLUSION: Our study revealed that the fishers possess information on the ecological aspects of local ornamental fish species, many of which are consistent with scientific literature. The information presented may assist in the decision-making process for the management of local fishery resources and contribute to the resumption of growth and sustainability in the capture of ornamental fish.

Qualidade microbiológica da água e dos mexilhões Perna perna (Linnaeus, 1758) cultivados em Piúma, Espírito Santo, Brasil / Microbiological quality of water and Perna perna (Linnaeus, 1758) mussels cultivated in Piúma, Espírito Santo, Brazil

RESUMO Este estudo objetivou verificar a qualidade microbiológica da água e dos mexilhões cultivados pela Associação dos Maricultores de Piúma (AMPI), Espírito Santo, Brasil. Foram realizadas sete coletas de água e mexilhões, mensalmente, entre outubro de 2016 e maio de 2017. Os mexilhões foram coletados nos long lines da AMPI, e em cada mês foram coletados 40 mexilhões Perna perna e 100 mL de água do local. O material coletado foi destinado ao laboratório para a realização das análises microbiológicas em duplicata, número mais provável de coliformes totais (CT) e termotolerantes (Ctt), presença ou ausência de Salmonella sp e número de unidades formadoras de colônias de Staphylococcus aureus. Os resultados mostraram que o número de Ctt nas amostras de água no mês de janeiro estavam acima do permitido pela Resolução n° 357 do Conselho Nacional do Meio Ambiente (CONAMA). Já os níveis de Ctt e Staphylococcus aureus na carne dos mexilhões mostraram-se dentro do limite aceitável pela RDC n° 12 da Agência Nacional de Vigilância Sanitária (ANVISA). Entretanto, foram encontradas bactérias com características do gênero Salmonella sp nos meses de dezembro e março nos mexilhões, impossibilitando sua comercialização e seu consumo. Durante esses meses, a cidade tem alto fluxo de turistas. Por fim, recomenda-se a realização das análises microbiológicas continuamente, principalmente no período do verão, época que tem grande fluxo de turistas no município de Piúma e que registrou presença de Salmonella na carne dos mexilhões e níveis de Ctt na água acima do permitido pelas legislações vigentes.

ABSTRACT This study aimed to verify the microbiological quality of water and mussels cultivated by the Piúma Farmers Association (Associação dos Maricultores de Piúma — AMPI), Espírito Santo, Brazil. Seven samples of water and mussels were collected monthly from October 2016 to May 2017. The mussels were collected from the long lines of the AMPI, and in each month 40 mussels Perna perna and 100 mL of water were sampled from the site. The collected material was sent to the laboratory for duplicate microbiological analysis, Most Probable Number of Total (CT) and Thermotolerant (Ctt) coliforms, presence or absence of Salmonella sp and number of Staphylococcus aureus colony forming units. The results showed that the number of Ctt in the water samples in January was higher than that allowed by Resolution 357 of the National Environment Council (Conselho Nacional do Meio Ambiente — CONAMA). The levels of Ctt and Staphylococcus aureus in mussel meat were within the acceptable range by the Brazilian National Environment Council (Agência Nacional de Vigilância Sanitária — ANVISA) Resolution RDC No. 12. However, colonies with characteristics of Salmonella sp were found in December and March in the mussels, making it impossible to sell and consume. This period coincides with a high flow of tourists in the municipality. Finally, it is recommended to perform microbiological analyzes continuously, especially in the summer, where there are a lot of tourists in the city of Piúma, period that showed the presence of Salmonella in the meat of mussels and Ctt levels in water above the allowed current legislation.

Toxicity of Copper in Epinephelus marginatus (Perciformes; Serranidae) After Ingestion of Contaminated Food.

The dusky grouper Epinephelus marginatus is predator fish subjected to be impacted due to the contamination of their habitats. A viable source of metal contamination, i.e., copper (Cu), in this species is the ingestion of contaminated food. The objective of this work was to verify the toxic effects of Cu contaminated feed in dusky grouper. A 15 days trial was conducted with three treatments: control, 1 g and 2 g Cu/kg of fish feed. After the trial, the gut was analyzed for Cu concentration and the liver for SOD and GST activity. The Cu concentration in the intestinal tract was significantly greater in fish from contaminated treatments when compared with control. The SOD was significantly lower in contaminated fish, and the GST did not show differences among treatments. Copper showed to be toxic for the species, as evidenced by gut accumulation and suggested by SOD response.

Efeito da quantidade de proteína na dieta e treinamento físico sobre parâmetros fisiológicos e zootécnicos de matrinchã (Brycon amazonicus, Günther 1869) / Dietary protein and exercise training effects on the physiological and zootechnical parameters of matrinchã (Brycon amazonicus, Günther 1869)

The aim of this study was to evaluate the combined effects of physical training and the dietary protein level on the hematological parameters, body proteins, lipids and ash content, food intake, conversion efficiency and growth ratio together with the swimming performance and the resistance to hypoxic stress in specimens of matrinchã. The results indicate that the swimming performance and hematology are not altered by physical training or by the diet protein content; moreover, the resistance to hypoxia is increased by the exercise, as indicated by the lower levels of plasma glucose. Increased amount of protein in the diet and training improve feed conversion and mass gain in the animals, while enhanced food intake is related to training. The simultaneous application of those two factors causes a combined and improved effect on the mass gain and the conversion rates. Due to higher fat accumulation in the fillet, future assessment of stored fat type is suggested, as well as evaluation of market acceptance of the fish product.

O objetivo do presente estudo foi avaliar a interação entre os efeitos do treinamento físico e da quantidade de proteína na dieta sobre a hematologia; a composição corpórea de proteínas, lipídeos e cinzas; o consumo alimentar, o ganho de massa e fator de conversão; o desempenho natatório e a resistência ao estresse causado pela hipóxia em exemplares de matrinchã. Os resultados indicam que a hematologia e o desempenho natatório não são alterados pelo treinamento físico ou pela concentração de proteína na dieta, porém, a resistência à hipóxia é aumentada pelo treinamento, como indicado por menores elevações dos níveis de glicose. O aumento da quantidade de proteína na dieta e o treinamento melhoram a conversão alimentar e o ganho de massa dos animais, enquanto que o aumento na ingestão de ração é causado apenas pelo treinamento. A aplicação concomitante dos dois fatores, embora cause efeito aditivo nos índices de ganho de massa e conversão alimentar, gera também os maiores acúmulos de gorduras no filé, o que sugere uma avaliação do tipo de gordura acumulada e da aceitação desta carne no mercado.

Tambaqui growth and survival when exposed to different photoperiods

The use of different photoperiods (light) were investigated during tambaqui (Colossoma macropomum) juvenile growth under captivity. Light intensity tested was: continuous dark (24hrs without light), natural photoperiod simulation (10hrs of light and 14hrs without light) and continuous light (24 with light). No mortality was recorded among treatments. Significant differences was observed after 50 days of experiment among mean fish weight, fish kept under a continuous darkness showed a better specific growth rate (6.02 percent) when compared to control fish (natural photo period, 3.67 percent). Fish exposed to continuous light presented the lowest mean specific growth rate (2.04 percent). It is possible to improve tambaqui juvenile weight gain performance when kept under continuous darkness.

Investigou-se o uso de diferentes fotos períodos (iluminação) durante o crescimento de juvenis de tambaqui (Colossoma macropomum) em cativeiro. As intensidades testadas foram: escuro contínuo (24hs sem luz), simulação do foto período natural (10hs de luz e 14hs sem luz) e iluminação contínua (24hs de luz). Não houve mortalidade nos diferentes tratamentos. Diferença significativa entre as médias do peso foi observada após 50 dias de experimento, a exposição ao escuro contínuo apresentou o melhor índice de crescimento específico (6,02 por cento) em relação ao controle (foto período natural, com 3,67 por cento). O menor índice de crescimento médio foi observado na exposição à iluminação contínua (2,04 por cento). Assim, pode-se melhorar o desempenho de ganho de peso para juvenis de tambaqui quando mantidos no escuro contínuo.