Ocimum gratissimum essential oil in the transport water of Brycon hilarii: implications at water quality, blood parameters and residues in tissue and plasma.

Transporting live fish is a common practice in fish farming, and is certainly one of the main problems that affect fish homeostasis. In this scenario, the use of natural additives has shown promise in improving fish resistance to adverse situations. This study aimed to assess the impact of Ocimum gratissimum L. essential oil (OGEO) on water quality, hematological parameters, and residue levels in the plasma, fillet, and liver of juvenile piraputanga (Brycon hilarii) during a two-hour transportation period. The fish were divided into plastic bags (4 L) and exposed to three different OGEO concentrations (10, 20, and 30 mg L-1), while a control group received no OGEO (three repetitions each). After the two-hour transportation, blood samples were collected, as well as portions of the fillet and liver for quantifying essential oil compounds, which were also measured in the plasma. Oxygen levels remained high throughout the transportation period, in all groups, while the pH decreased. Hemoglobin, MCHC, and MCH increased in fish exposed to OGEO concentrations of 20 and 30 mg L-1, compared to the control group. However, lymphocyte counts and the concentrations of essential oil compounds in plasma, fillet, and liver increased with higher OGEO concentrations. The use of 10 mg L-1 OGEO in the two-hour transport water is promising to ensure the survival and well-being of Brycon hilarii juveniles (weighing 16 g), showing to be safe and effective. The residual concentration of eugenol the major compound of OGEO in the fillet remains below the maximum limit of the recommended daily intake.

Hematological, biochemical and oxidative responses induced by thermal shock in juvenile Tambaqui (Colossoma macropomum) and its hybrid Tambatinga (Colossoma macropomum x Piaractus brachypomus).

The effects of thermal shock on hematological, biochemical and antioxidant responses were evaluated in liver tissue of juvenile tambaqui (Colossoma macropomum) and tambatinga (♀ C. macropomum × â™‚ Piaractus brachypomus). Forty juveniles of tambaqui and 40 juveniles of tambatinga, of the same age and with an initial weight of 23.3 ± 6.7 g, were randomly distributed in eight 28L circular tanks. A tank (n = 10 fish) of tambaqui and a tank (n = 10 fish) of tambatinga were then used to obtain basal data. The other animals were subjected to thermal shock with sudden temperature reduction from 28 to 18 ºC. Blood and tissue were then collected after 1, 6 and 24 h from the onset of thermal shock. No mortality was observed during the experimental period. Thermal shock increased triglyceride levels after 24 h of stress for tambaqui and reduced values for tambatinga. There was an effect on plasma glucose only for fish group (P < 0.0001) and collection time (P < 0.0001) with a peak observed for the hybrid after 6 h. The interaction of factors for SOD indicated greater activity for tambatinga at the 6 h collection and lower at basal and 1 h collections. There was an interaction for CAT (P = 0.0020) with less activity for tambatinga at 1 h. However, thermal shock and hybridization did not influence GST and TBARS levels in liver tissue. Therefore, the results suggest that the hybrid, tambatinga, is more efficient at promoting adjustments of biochemical responses and antioxidant enzymes during thermal shock.

Morphological and cytochemical characterization of the peripheral blood cells of farmed streaked prochilod Prochilodus lineatus (Characiformes, Prochilodontidae) / [Caracterização morfológica e citoquímica das células do sangue periférico do curimatã Prochilodus lineatus (Characiformes, Prochilodontidae) em cultivo]

Morphological and cytochemical studies of peripheral blood cells of fish have improved the understanding of their functions and cell types. The present study performed the Morphological and cytochemical analysis of the peripheral blood of Prochilodus lineatus, Characiform native to South America, which has been gaining space in local aquaculture and as a species introduced in Asia. Our analysis provided information on the morphological and cytochemical characteristics of the leukocytes, for the formulation of hypothesis about their role in the immune system of the species. It was found that Prochilodus lineatus has morphological and cytochemical features in common with other fish species, mainly of the Characiformes order. However, we detected the presence of heterophils and PAS positive granulocytes simultaneously with neutrophils. We also found that heterophils and PAS positive granulocytes are very similar, both morphologically and cytochemically.(AU)

O estudo da morfologia e da citoquímica das células do sangue periférico dos peixes tem sido eficaz para o entendimento de suas funções e dos tipos celulares. Este estudo realizou a análise morfológica e citoquímica do sangue periférico de Prochilodus lineatus, caracídeo nativo da América do Sul que vem ganhando espaço na aquicultura local e como espécie introduzida na Ásia. Essa análise forneceu informações sobre a morfologia e as características citoquímicas dos leucócitos, visando a hipóteses sobre suas funções. Verificou-se que estas são semelhantes em vários aspectos a outras espécies, principalmente da ordem Characiformes. No entanto, neste estudo detectou-se a presença dos heterofilos e da célula granulocítica especial, simultaneamente à presença dos neutrófilos. Ainda, foi verificado que os heterofilos e a célula granulocítica especial são muito semelhantes morfológica e citoquimicamente.(AU)

Low temperature stress in a cultured fish (Piaractus mesopotamicus) fed with Pyropia columbina red seaweed-supplemented diet.

This study aimed to analyze the cold stress effects (in terms of hematology, energy reserves, and oxidative stress) in Piaractus mesopotamicus (pacú) and their mitigation by a Pyropia columbina red seaweed-supplemented diet. For this purpose, juvenile fish were fed with a control (CD) or a red seaweed-supplemented diet (RD) for 60 days, and then, the animals were exposed to a low temperature (14 °C) and a control temperature (24 °C) for 24 h. The cold shock generated an increase of hemoglobin levels in fish fed with both diets. In CD-fed fish, plasmatic triglycerides, cholesterol, and hepatic glycogen decreased after the thermal shock; meanwhile, the animals fed with RD showed decreased hepatic proteins, but increased cholesterol and hepatic glycogen. Regarding oxidative stress, antioxidant enzymes augmented their activity in the liver, intestine, and gills; meanwhile, lipid oxidative damage was observed in the liver and intestine of fish exposed to 14 °C and fed with both diets. Pacú was sensitive to cold shock, but no mitigation effects were observed in fish fed with the supplemented diet. Further research should target higher concentrations of P. columbina in supplemented diets to take advantage of this valuable resource.

Modulation of the innate immune response, antioxidant system and oxidative stress during acute and chronic stress in pacu (Piaractus mesopotamicus).

Stress is an energy-demanding process, as well as the responses of the innate immune system, that impose a metabolic overload on cellular energy production, which can affect the cellular redox balance, causing oxidative damage. We evaluated the role of stress in the modulation of innate immune and oxidative/antioxidant mechanisms in juvenile pacu exposed to acute and chronic stressors. The experimental period lasted 30 days, and fish (113.7 ± 35.1 g) were fed commercial feed. During this period, half of the fish were not manipulated (Condition A), and the other half were chased with a dip net for 5 min twice a day (Condition C). After the 30-day period, fish from both groups were sampled (baseline sampling), and the remainders (not sampled) were air exposed for 3 min (acute stressor), returned to the tanks, and were sampled again 30 min, 3 h, 6 h, and 24 h after air exposure. We evaluated biomarkers of stress (circulating cortisol and glucose), the innate immune system (respiratory burst activity/RBA, hemolytic activity of the complement system (HA-AP) and serum concentration of lysozyme), oxidative damage (lipid peroxidation/LPO), and antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; and glutathione peroxidase, GSH-Px). Our results showed that stress, acutely or chronically, caused a transient reduction of RAL and activated the HA-AP. Acutely, stress increased the lysozyme concentration. Furthermore, both conditions caused oxidative stress in the liver, and differently they modulated the antioxidant system, enhancing SOD activity and impairing CAT and GSH-Px activity.

Integrated use of biomarkers to evaluate the reproductive physiology of Astyanax fasciatus and Hoplias malabaricus males (Teleostei: Characiformes) in polluted reservoirs.

The reproductive physiology of fish can be changed by the presence of pollutants in the water, which act as endocrine disrupting compounds (EDC). We evaluated the impacts of water contaminants in polluted reservoirs acting as possible EDC on the reproductive physiology of Astyanax fasciatus and Hoplias malabaricus males. We used biomarkers with different levels of biological organization. Hoplias malabaricus adult males were collected in the summer and winter at five different sites in the Tietê River Basin: the Ponte Nova reservoir (PN), considered a reference site due to the low anthropogenic influence; the Billings reservoir (BIL) at two different branches; and the Guarapiranga reservoir (GUA) at two different branches. Astyanax fasciatus adult males were collected at PN and BIL. BIL and GUA are subjected to great anthropogenic action. We analyzed gonadal histomorphology, testosterone (T), 11-ketotestosterone (11-KT), estradiol (E2) plasma levels, and gene expression of hepatic vitellogenin (vtgA) and pituitary follicle stimulating hormone (fshß). In the PN reservoir (reference), the biomarkers analyzed in both species did not differ between the periods analyzed. This is an evidence that the animals keep the same reproductive activity during both seasons. The changes in the plasma concentration of gonadal steroids in both species in polluted reservoirs suggest the presence of EDC compounds in the water and/or adjusts of the physiological setpoint to allow the reproduction in such adverse conditions. The use of vtgA as biomarker suggests the presence of estrogenic compounds, mainly in BIL, but with a more evident response of H. malabaricus. However, even considering physiological changes, both species present testes during the maturation phase that allow the reproduction in an environment with a high degree of pollution.

Age-associated hemogram and ultrastructural leukocyte morphology in Pygocentrus nattereri (Characiformes: Serrasalmidae) from the Brazilian Pantanal

Pygocentrus nattereri is a carnivorous fish widely distributed in the Brazilian wetland and occupies different river strata. Based on the sexual maturity, the standard length (SL) has been used to categorize age and physiological attributes. However, hematological parameters and their relationship with age and growth features are unknow. Here, the aim was to evaluate the hematological profile and leukocyte data of juvenile and adult specimens, associating it with SL. Specimens of both sexes were weighted, and SL were measured for classify as juveniles (SL ≤16.0 cm) and adults (SL ≥16.1 cm). Blood was collected to perform the hemogram, for leukocyte quantification and ultrastructural descriptions. In general, hemogram values and leukocyte, lymphocyte, and monocyte counts were high in juveniles. Hemogram variables were negatively correlated with SL. Thrombocyte and neutrophil counts were higher in adults than in juveniles. SL was correlated with leukogram variables. Ultrastructural analysis showed that granular leukocyte count in P. nattereri was similar another fishes. PAS-positive granular leukocytes are slightly smaller than neutrophils and are characterized by dense elongated and semi-lunar shapes in the cytoplasm. Our results suggest that age-based SL may be useful for the analysis of the relationship between P. nattereri health status and wetland aquatic environments.(AU)

Pygocentrus nattereri é um peixe carnívoro amplamente distribuído no pantanal brasileiro e ocupa diferentes estratos de rios. Com base na maturidade sexual, o comprimento padrão (CP) tem sido usado para categorizar a idade e os atributos fisiológicos. No entanto, os parâmetros hematológicos e sua relação com a idade e características de crescimento são desconhecidos. O objetivo foi avaliar o perfil hematológico e leucocitário de espécimes juvenis e adultos, associando-o ao CP. Espécimes de ambos os sexos foram pesados e os CPs foram medidos para classificação em juvenis (CP ≤16,0 cm) e adultos (CP ≥16,1 cm). Foi coletado sangue para realização do hemograma, perfil e morfologia dos leucócitos. Em geral, os valores do hemograma e as contagens de leucócitos, linfócitos e monócitos foram maiores nos jovens. A contagem de trombócitos e neutrófilos foi maior em adultos do que em jovens. A análise ultraestrutural mostrou que a contagem de leucócitos granulares em P. nattereri foi semelhante a outros peixes. Os leucócitos granulares PAS-positivos são menores do que os neutrófilos, sendo caracterizados por formas alongadas e semilunares densas no citoplasma. Nossos resultados sugerem que CP é útil para a análise da relação entre o estado de saúde de P. nattereri em ambientes aquáticos alagados.(AU)

Age-associated hemogram and ultrastructural leukocyte morphology in Pygocentrus nattereri (Characiformes: Serrasalmidae) from the Brazilian Pantanal

Pygocentrus nattereri is a carnivorous fish widely distributed in the Brazilian wetland and occupies different river strata. Based on the sexual maturity, the standard length (SL) has been used to categorize age and physiological attributes. However, hematological parameters and their relationship with age and growth features are unknow. Here, the aim was to evaluate the hematological profile and leukocyte data of juvenile and adult specimens, associating it with SL. Specimens of both sexes were weighted, and SL were measured for classify as juveniles (SL ≤16.0 cm) and adults (SL ≥16.1 cm). Blood was collected to perform the hemogram, for leukocyte quantification and ultrastructural descriptions. In general, hemogram values and leukocyte, lymphocyte, and monocyte counts were high in juveniles. Hemogram variables were negatively correlated with SL. Thrombocyte and neutrophil counts were higher in adults than in juveniles. SL was correlated with leukogram variables. Ultrastructural analysis showed that granular leukocyte count in P. nattereri was similar another fishes. PAS-positive granular leukocytes are slightly smaller than neutrophils and are characterized by dense elongated and semi-lunar shapes in the cytoplasm. Our results suggest that age-based SL may be useful for the analysis of the relationship between P. nattereri health status and wetland aquatic environments.(AU)

Pygocentrus nattereri é um peixe carnívoro amplamente distribuído no pantanal brasileiro e ocupa diferentes estratos de rios. Com base na maturidade sexual, o comprimento padrão (CP) tem sido usado para categorizar a idade e os atributos fisiológicos. No entanto, os parâmetros hematológicos e sua relação com a idade e características de crescimento são desconhecidos. O objetivo foi avaliar o perfil hematológico e leucocitário de espécimes juvenis e adultos, associando-o ao CP. Espécimes de ambos os sexos foram pesados e os CPs foram medidos para classificação em juvenis (CP ≤16,0 cm) e adultos (CP ≥16,1 cm). Foi coletado sangue para realização do hemograma, perfil e morfologia dos leucócitos. Em geral, os valores do hemograma e as contagens de leucócitos, linfócitos e monócitos foram maiores nos jovens. A contagem de trombócitos e neutrófilos foi maior em adultos do que em jovens. A análise ultraestrutural mostrou que a contagem de leucócitos granulares em P. nattereri foi semelhante a outros peixes. Os leucócitos granulares PAS-positivos são menores do que os neutrófilos, sendo caracterizados por formas alongadas e semilunares densas no citoplasma. Nossos resultados sugerem que CP é útil para a análise da relação entre o estado de saúde de P. nattereri em ambientes aquáticos alagados.(AU)

Physiological and metabolic responses in juvenile Colossoma macropomum exposed to hypoxia.

This study aimed to evaluate hematological, biochemical, and gasometric parameters of tambaqui juveniles (Colossoma macropomum) exposed to hypoxia and subsequent recovery. Six animals were subjected to normoxia (basal) treatment with dissolved oxygen (DO) 6.27 ± 0.42 mg L-1. Water flow and aeration were reduced for 3 days (hypoxia), during which DO was 0.92 ± 0.37 mg L-1. Water flow and aeration were then reestablished with DO remaining similar to basal. The treatments were as follows: normoxia (basal); 24 h after initiating hypoxia (24H); 72 h after initiating hypoxia (72H); 24 h after reestablishing normoxia (24R); 48 h after reestablishing normoxia (48R); and 96 after reestablishing normoxia (96R). The highest glucose level was recorded at 24H (P < 0.05); the highest lactate level was at 72R; and the highest blood pH was at 24H and 72H (P < 0.05). The highest concentration of PvCO2 was at 24H (P < 0.05), while at 96R it was equivalent to basal (P > 0.05). The variable PvO2 was only higher than basal at 24R (P < 0.05). Juvenile C. macropomum managed to reestablish the main stress indicators (glucose and lactate) at 96R, while the other indicators varied during the study, with homeostatic physiology being reestablished during the recovery period.

Hematological profile of Astyanax bimaculatus under different offer of Lactobacillus sp / Perfil hematológico de Astyanax bimaculatus sob diferentes ofertas do Lactobacillus sp

The objective of this study was to evaluate the hematological and immunological parameters of yellowtail lambaris (Astyanax bimaculatus), fed with different frequencies of the probiotic (Lactobacillus spp.). Fishes were distributed into 20 experimental units and divided in five treatments: control (0%), 25%, 50%, 75% and 100% of probiotic supply. A higher presence of total leukocytes (47.70 103cell µl-1), lymphocytes (36.11 103cell µl-1) and monocytes (11.58 103cell µl-1) was verified in fish fed 100% of probiotic, showing a directly proportional ratio among the frequencies of the probiotic supply on the availability of circulating cells in the circulatory system (R² 094-0.97). Hematocrit (27.30-34.63%), hemoglobin (7.00-10.90g dl-1), mean corpuscular volume (4.21-5.45 10-5.pg), mean corpuscular hemoglobin (3.45-5.40 10-6.pg), mean corpuscular hemoglobin concentration, (2.99-4.35g dl-1), total protein (44.32-50.26mg ml-1) and total plasma immunoglobulin (27.96-34.08mg ml-1) did not diverge among treatments. The frequency of the probiotic supply interferes with the hematological profile, although lactic acid bacteria were present in the same concentrations in the intestinal tract, regardless of the probiotic supply, there was an increase in circulating leukocytes, especially lymphocytes and monocytes, in lambari fed probiotic with more frequency.(AU)

O objetivo do presente trabalho foi avaliar os parâmetros hematológicos e imunológicos do lambari-do-rabo-amarelo (Astyanax bimaculatus) alimentado com diferentes frequências de probiótico (Lactobacillus spp.). Os peixes foram distribuídos em 20 unidades experimentais e divididos em cinco tratamentos: controle (0%), 25%, 50%, 75% e 100% de frequência na suplementação probiótica. Alta presença de leucócitos totais (47,70 10³ células µ-1), linfócitos (36,11 10³ células µ-1) e monócitos (11,58 10³ células µ-1) em peixes alimentados com 100% de probiótico apresenta uma taxa diretamente proporcional entre as frequências da suplementação probiótica na disponibilidade das células no sistema circulatório (R² 094-0,97). Hematócrito (27,30-34,63%), hemoglobina (7,00-10,90g dL-¹), volume corpuscular médio (4,21-5,45 10-5.pg), hemoglobina corpuscular média (3,45-5,40 10-6.pg), concentração de hemoglobina corpuscular média (2,99-4,35g dl-1), proteína total (44,32-50,26mg ml-1) e imunoglobulina plasmática total (27,96-34,08mg ml-1) não divergiram entre os tratamentos. A frequência da suplementação probiótica interferiu no perfil hematológico. Embora as bactérias ácido láticas estejam presentes na mesma concentração no trato intestinal, independentemente da oferta de probiótico, houve um aumento na circulação de leucócitos, especialmente linfócitos e monócitos, nos lambaris alimentados com maior frequência.(AU)

Hematological profile of Astyanax bimaculatus under different offer of Lactobacillus sp / Perfil hematológico de Astyanax bimaculatus sob diferentes ofertas do Lactobacillus sp

The objective of this study was to evaluate the hematological and immunological parameters of yellowtail lambaris (Astyanax bimaculatus), fed with different frequencies of the probiotic (Lactobacillus spp.). Fishes were distributed into 20 experimental units and divided in five treatments: control (0%), 25%, 50%, 75% and 100% of probiotic supply. A higher presence of total leukocytes (47.70 103cell µl-1), lymphocytes (36.11 103cell µl-1) and monocytes (11.58 103cell µl-1) was verified in fish fed 100% of probiotic, showing a directly proportional ratio among the frequencies of the probiotic supply on the availability of circulating cells in the circulatory system (R² 094-0.97). Hematocrit (27.30-34.63%), hemoglobin (7.00-10.90g dl-1), mean corpuscular volume (4.21-5.45 10-5.pg), mean corpuscular hemoglobin (3.45-5.40 10-6.pg), mean corpuscular hemoglobin concentration, (2.99-4.35g dl-1), total protein (44.32-50.26mg ml-1) and total plasma immunoglobulin (27.96-34.08mg ml-1) did not diverge among treatments. The frequency of the probiotic supply interferes with the hematological profile, although lactic acid bacteria were present in the same concentrations in the intestinal tract, regardless of the probiotic supply, there was an increase in circulating leukocytes, especially lymphocytes and monocytes, in lambari fed probiotic with more frequency.(AU)

O objetivo do presente trabalho foi avaliar os parâmetros hematológicos e imunológicos do lambari-do-rabo-amarelo (Astyanax bimaculatus) alimentado com diferentes frequências de probiótico (Lactobacillus spp.). Os peixes foram distribuídos em 20 unidades experimentais e divididos em cinco tratamentos: controle (0%), 25%, 50%, 75% e 100% de frequência na suplementação probiótica. Alta presença de leucócitos totais (47,70 10³ células µ-1), linfócitos (36,11 10³ células µ-1) e monócitos (11,58 10³ células µ-1) em peixes alimentados com 100% de probiótico apresenta uma taxa diretamente proporcional entre as frequências da suplementação probiótica na disponibilidade das células no sistema circulatório (R² 094-0,97). Hematócrito (27,30-34,63%), hemoglobina (7,00-10,90g dL-¹), volume corpuscular médio (4,21-5,45 10-5.pg), hemoglobina corpuscular média (3,45-5,40 10-6.pg), concentração de hemoglobina corpuscular média (2,99-4,35g dl-1), proteína total (44,32-50,26mg ml-1) e imunoglobulina plasmática total (27,96-34,08mg ml-1) não divergiram entre os tratamentos. A frequência da suplementação probiótica interferiu no perfil hematológico. Embora as bactérias ácido láticas estejam presentes na mesma concentração no trato intestinal, independentemente da oferta de probiótico, houve um aumento na circulação de leucócitos, especialmente linfócitos e monócitos, nos lambaris alimentados com maior frequência.(AU)

Under stress conditions, pacu Piaractus mesopotamicus modulates the metabolic allostatic load even after Dolops carvalhoi challenge to maintain self-protection mechanisms.

Fish metabolic allostatic dynamics, when animal present physiological modifications that can be strategies to survive, are important for promoting changes to ensure whole body self-protection and survival in chronic states of stress. To determine the impact of sequential stressors on pacu (Piaractus mesopotamicus), fish were subjected to two trials of stressful treatments, administration of exogenous dietary cortisol, and parasite challenge. The first experiment consisted of a two-day acute stress trial and the second, an eight-day chronic stress trial, and after both experiments, fish parasite susceptibility was assessed with the ectoparasite Dolops carvalhoi challenge. Physiological changes in response to acute trial were observed in glycogen, cortisol, glucose, osmolarity, sodium, calcium, chloride, potassium, hematocrit, hemoglobin, red blood cells and mean corpuscular volume, and white blood cell (P < 0.05), whereas response to chronic trial were observed in glycogen, osmolarity, potassium, calcium, chloride, mean corpuscular volume, white blood cell, neutrophil, and lymphocyte (P < 0.05). Acute trials caused physiological changes, however those changes did not induce the consumption of hepatic glycogen. Chronic stress caused physiological changes that induced hepatic glycogen consumption. Under acute trial, stress experience was important to fish to achieve homeostasis after chronic stress. Changes were important to modulate the response to stressor, improve body health status, and overcome the extra stressor with D. carvalhoi challenge. The experiments demonstrate that pacu initiate strategic self-protective metabolic dynamics in acute states of stress that ensure the maintenance of important life processes in front of sequential stressors.

In vivo phagocytosis and hematology in Astyanax altiparanae, a potential model for surrogate technology

Although the potential of surrogate propagation technology for aquaculture and conservation of Neotropical fish, the poor understanding of the host immune system may results in rejection and destruction of the donor material. Thus, it is necessary to study and to develop methods to evaluate the effects of immunosuppressive drugs employment and to evaluate the immunocompatibility between donor and receptor. Thus, the present study aimed to optimize a methodology to assess in vivo phagocytosis in Astyanax altiparanae using Saccharomyces cerevisiae and to evaluate their hematological response resultant from the inflammatory induction. To this, S. cerevisiae were labeled with Congo red and injected in the coelomic cavity of A. altiparanae at the concentration of 2.5 x 106 cells mL-1. A PBS solution and a non-injected group were kept as control. Fish blood was sampled and the phagocytic capacity and index were determined at 1, 2, 3 and 6 h post-injection (hpi). The yeast injection successfully stimulated phagocytosis, with the best result for phagocytosis assessment after 2 hpi. Moreover, it was achieved a high traceability of phagocytized and non-phagocytized yeast under optic microscopy analysis due to the Congo red labeling. The hematological profile was similar to usually observed in early infections, indicating lymphocyte migration to inflammatory site and increase in number of circulating phagocytes due to natural response to inflammatory stimulus. In conclusion, our method was efficient to assess in vivo phagocytosis in A. altiparanae and will be an important tool to evaluate the efficacy of immunosuppressive drugs in this species. Additionally, these results may serve as support for further studies in fish immunocompetence, both in laboratory and in field conditions.(AU)

Apesar do potencial apresentado pela tecnologia de propagação mediada para a aquicultura e conservação de peixes Neotropicais, o pobre entendimento do sistema imune do hospedeiro pode resultar na rejeição e destruição do material do doador. Com isso, se fazem necessários o estudo e o desenvolvimento de métodos para análise tanto dos efeitos de drogas imunossupressoras quanto para a avaliação da imunocompatibilidade entre doadores e receptores. Logo, o presente estudo teve como objetivo aperfeiçoar um método para analisar a fagocitose in vivo em Astyanax altiparanae usando Saccharomyces cerevisiae marcado e avaliar seu perfil hematológico resultante da indução inflamatória. Para isso, S. cerevisiae foram marcados com vermelho Congo e injetados na cavidade celomática dos A. altiparanae na concentração de 2,5 x 106 células.mL-1. Peixes injetados com PBS e peixes não injetados foram mantidos como controle. Sangue foi colhido e a capacidade fagocítica e o índice fagocítico foram determinados após 1, 2, 3 e 6 horas após à injeção (hpi). A injeção de levedura estimulou a fagocitose com sucesso, com o melhor resultado atingido após 2 hpi. Ainda, foi observada uma alta rastreabilidade das leveduras fagocitadas e não fagocitadas sob microscopia óptica devido à marcação com vermelho Congo. O perfil hematológico foi similar ao observado usualmente em infecções recém-induzidas, indicando migração de linfócitos ao sítio inflamatório e aumento no número de fagócitos circulantes devido à resposta natural ao estímulo inflamatório. Como conclusão, nosso método foi eficiente para analisar a fagocitose in vivo em A. altiparanae e será uma ferramenta importante para a avaliação de eficácia de drogas imunossopressoras para esta espécie. Em adição, estes resultados podem contribuir para futuros estudos em imunocompetência em peixes, tanto em âmbito laboratorial quanto a campo.(AU)

Osmoregulatory disturbance in Neotropical fish exposed to the crude extracts of the cyanobacterium, Radiocystis fernandoi.

Blooms of cyanobacteria, a common event in eutrophic environments, result in the release of potentially toxic substances into the water. The cyanobacterium Radiocystis fernandoi produces microcystin (MC) and other peptides that may disturb homeostasis. This study evaluated the effect of intraperitoneal injections containing the crude extract (CE) of R. fernandoi strain R28 on the gills and kidneys of neotropical fish, Piaractus mesopotamicus, 3, 6 and 24 h post-injection. CE contained MC-RR, MC-YR and minor other oligopeptides. Plasma ions and the activities of the enzymes PP1 and PP2A, Na+/K+-ATPase (NKA), H+-ATPase (HA) and carbonic anhydrase (CA) were determined and morphological changes in both the gills and kidneys were characterized. Compared to controls, the concentration of Na+ within the plasma of P. mesopotamicus decreased after treatment with CE 3 h post treatment and increased after 24 h; the concentration of K+ decreased after 6 h. The activity of the endogenous PP1 and PP2A was unchanged in the gills and was inhibited in the kidneys 6 h after i.p. injection. In the gills, NKA activity increased after 3 h and decreased 6 h post i.p. exposure. Further, NKA activity did not differ from the controls 24-h post injection. In the kidneys, NKA, HA and CA activities were unaffected by treatment. The mitochondria-rich cell (MRC) density in the gills decreased after 3 h in the filament and 3 and 6 h in the lamellae and was restored to the control levels 24 h post-exposure. Filament epithelial hyperplasia and hypertrophy, lamellar atrophy and rupture of the lamellar epithelium were the most common effects of treatment in the gills. No histopathological changes occurred in the kidneys. This study demonstrates that a single dose of toxic CE from R. fernandoi can cause a transitory ion imbalance in P. mesopotamicus which is related to the changes in MRC levels and NKA activity. Ionic balance was recovered 24 h post i.p. injection, however, morphological changes that occurred in the gills took a longer amount of time to return to normal. To conclude, the effects of components contained within the CE of R. fernandoi may be harmful to P. mesopotamicus. In particular, the recovery of ionic regulation depends on MRC responses and histopathological changes produced by CE may affect gas exchange and other gill functions.

Overview of the toxic effects of titanium dioxide nanoparticles in blood, liver, muscles, and brain of a Neotropical detritivorous fish.

The toxicity of titanium dioxide nanoparticles (TiO2 -NP) in the blood, liver, muscle, and brain of a Neotropical detritivorous fish, Prochilodus lineatus, was tested. Juvenile fish were exposed to 0, 1, 5, 10, and 50 mg L-1 of TiO2 -NP for 48 hours (acute exposure) or 14 days (subchronic exposure) to evaluate changes in hematology, red blood cell (RBC) genotoxicity/mutagenicity, liver function (reactive oxygen species (ROS) production, antioxidant responses, detoxification, and histopathology), acetylcholinesterase (AChE) activity in muscles and brain, and Ti bioaccumulation. TiO2 -NP did not cause genetic damage to RBC, but acutely decreased white blood cells (WBC) and increased monocytes. Subchronically, RBC decreased, mean cell volume and hemoglobin increased, and WBC and lymphocytes decreased. Therefore, NP has the potential to affect immune system and increase energy expenditure, reducing the fish's ability to avoid predator and to resist pathogens. In the liver, acute exposure decreased ROS and increased glutathione (GSH) content, while subchronic exposure decreased superoxide dismutase activity and increased glutathione-S-transferase (GST) activity and GSH content. GSH and GST seem to play an essential role in metabolizing NP and ROS, likely increasing hepatocytes' metabolic rate, which may be the cause of observed cell hypertrophy, disarrangement of hepatic cords and degenerative morphological alterations. Although most studies indicate that the kidney is responsible for metabolizing and/or eliminating TiO2 -NP, this study shows that the liver also has a main role in these processes. Nevertheless, Ti still accumulated in the liver, muscle, and brain and decreased muscular AChE activity after acute exposure, showing neurotoxic potential. More studies are needed to better understand the biochemical pathways TiO2 -NP are metabolized and how its bioaccumulation may affect fish homeostasis and survival in the environment.

Hematological parameters of the hybrid serrasalmids farmed in Central-Western Brazil / Parâmetros hematológicos de serrasalmídeos híbridos cultivados na região Centro-Oeste do Brasil

This study evaluated the hematological parameters of tambacu and patinga, two the hybrid serrasalmid fishes farmed in the state of Mato Grosso do Sul, Brazil, with emphasis on the influence of seasons on them. Two-hundred forty hybrid fishes, 120 tambacus and 120 patingas, were collected over 12 months to assess their hematological parameters and compare them between the warm (October-March) and cold seasons (April-September). Water quality parameters were also measured weekly to test for the influence of environmental conditions on the results. The dissolved oxygen content in the water decreased in the warm season, and increased in the cold season. In the warm season, tambacus presented significantly higher values (p<0.05) of the total plasmatic protein (TPP), hemoglobin, mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and eosinophil count parameters. In the cold season, counts of erythrocytes, thrombocytes, total leukocytes, lymphocytes, and neutrophils were higher in tambacus (p<0.05). For patinga, MCV was higher in the warm season, while hemoglobin, erythrocytes, MCHC, total leukocytes, and basophil counts were higher (p<0.05) in the cold season. These results demonstrated the influences of seasonality on the hematological parameters of the hybrids tambacu and patinga when held under normal farming conditions in the region.

Este estudo avaliou os parâmetros hematológicos de híbridos tambacu e patinga cultivados no Mato Grosso do Sul, Brasil, com ênfase na influência das épocas do ano. Duzentos e quarenta peixes híbridos, 120 tambacus e 120 patingas, foram coletados ao longo de 12 meses para análises hematológicas, comparando a estação quente (Outubro-Março) com a estação fria (Abril-Setembro). Os parâmetros de qualidade de água foram medidos semanalmente, para verificar sua influência sobre os parâmetros hematológicos. A estação quente mostrou tendência ao declínio nos níveis de oxigênio dissolvido, enquanto na estação fria ocorreu o inverso. Na estação quente, tambacus apresentaram valores mais altos (p<0,05) de proteína plasmática total (TPP), concentração de hemoglobina, volume corpuscular médio (MCV), concentração de hemoglobina corpuscular média (MCHC) e número de eosinófilos. Na estação fria, as contagens de eritrócitos, trombócitos, leucócitos totais, linfócitos e neutrófilos foram mais altas (p<0,05). Em patingas, valores mais altos de MCV foram observados na estação quente, enquanto a concentração de hemoglobina, eritrócitos, MCHC, leucócitos totais e basófilos foram mais altos (p<0,05) na estação fria. Os resultados mostram influência da sazonalidade sobre os parâmetros hematológicos dos híbridos tambacu e patinga mantidos em condições típicas de cultivo na região.

Hematological parameters of the hybrid serrasalmids farmed in Central-Western Brazil / Parâmetros hematológicos de serrasalmídeos híbridos cultivados na região Centro-Oeste do Brasil

This study evaluated the hematological parameters of tambacu and patinga, two the hybrid serrasalmid fishes farmed in the state of Mato Grosso do Sul, Brazil, with emphasis on the influence of seasons on them. Two-hundred forty hybrid fishes, 120 tambacus and 120 patingas, were collected over 12 months to assess their hematological parameters and compare them between the warm (October-March) and cold seasons (April-September). Water quality parameters were also measured weekly to test for the influence of environmental conditions on the results. The dissolved oxygen content in the water decreased in the warm season, and increased in the cold season. In the warm season, tambacus presented significantly higher values (p<0.05) of the total plasmatic protein (TPP), hemoglobin, mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and eosinophil count parameters. In the cold season, counts of erythrocytes, thrombocytes, total leukocytes, lymphocytes, and neutrophils were higher in tambacus (p<0.05). For patinga, MCV was higher in the warm season, while hemoglobin, erythrocytes, MCHC, total leukocytes, and basophil counts were higher (p<0.05) in the cold season. These results demonstrated the influences of seasonality on the hematological parameters of the hybrids tambacu and patinga when held under normal farming conditions in the region.(AU)

Este estudo avaliou os parâmetros hematológicos de híbridos tambacu e patinga cultivados no Mato Grosso do Sul, Brasil, com ênfase na influência das épocas do ano. Duzentos e quarenta peixes híbridos, 120 tambacus e 120 patingas, foram coletados ao longo de 12 meses para análises hematológicas, comparando a estação quente (Outubro-Março) com a estação fria (Abril-Setembro). Os parâmetros de qualidade de água foram medidos semanalmente, para verificar sua influência sobre os parâmetros hematológicos. A estação quente mostrou tendência ao declínio nos níveis de oxigênio dissolvido, enquanto na estação fria ocorreu o inverso. Na estação quente, tambacus apresentaram valores mais altos (p<0,05) de proteína plasmática total (TPP), concentração de hemoglobina, volume corpuscular médio (MCV), concentração de hemoglobina corpuscular média (MCHC) e número de eosinófilos. Na estação fria, as contagens de eritrócitos, trombócitos, leucócitos totais, linfócitos e neutrófilos foram mais altas (p<0,05). Em patingas, valores mais altos de MCV foram observados na estação quente, enquanto a concentração de hemoglobina, eritrócitos, MCHC, leucócitos totais e basófilos foram mais altos (p<0,05) na estação fria. Os resultados mostram influência da sazonalidade sobre os parâmetros hematológicos dos híbridos tambacu e patinga mantidos em condições típicas de cultivo na região.(AU)

Changes in hematological and biochemical parameters of tambaqui (Colossoma macropomum ) parasitized by metazoan species / Alterações nos parâmetros hematológicos e bioquímicos de tambaqui (Colossoma macropomum) parasitado por espécies de metazoários

Abstract The aim of this study was to evaluate the impacts of metazoan parasites on hematological and biochemical parameters and relative condition factor of tambaqui (Colossoma macropomum ) farmed in northern Brazil. A total of 32 juvenile fish were captured from a commercial fish farm located in the municipality of Rio Preto da Eva, Amazonas state, Brazil. Parasite prevalence was 100% for Anacanthorus spathulatus, Mymarothecium boegeri and Notozothecium janauachensis, 100% for Neoechinorhynchus buttnerae and 53.13% for Dolops geayi. The greatest mean parasite intensity was found in acantocephalans followed by monogeneans and branchiuran crustaceans. A negative correlation was observed between abundance of N. buttnerae and hematocrit percentage, hemoglobin concentration, total thrombocyte count and glucose and between abundance of the monogenean and glucose concentration. Parasitic infections caused damage in tambaqui in terms of the observed hematological parameters that were characterized by hypochromic anemia and thrombocytopenia, which are important parameters to be used in parasitic diagnosis. This study is the first record of the occurrence of Dolops geayi in farmed tambaqui in the Amazon.

Resumo O objetivo deste estudo foi avaliar o impacto do parasitismo por metazoários sobre os parâmetros hematológicos, bioquímicos e fator de condição relativo de tambaquis (Colossoma macropomum) cultivados na região Norte do Brasil. Foram coletados 32 tambaquis em fase de engorda numa piscicultura comercial no município de Rio Preto da Eva, estado do Amazonas, Brasil. A prevalência de parasitos foi de 100% para Anacanthorus spathulatus, Mymarothecium boegeri e Notozothecium janauachensis, 100% para Neoechinorhynchus buttnerae e 53,13% para Dolops geayi. A maior intensidade média foi de acantocéfalos, seguida por monogenéticos e branquiúros. Correlação negativa significativa foi observada entre abundância de N. buttnerae e os parâmetros de hematócrito, concentração de hemoglobina, trombócitos totais e glicose, bem como entre a abundância de monogenéticos e glicose. A infecção por parasitos metazoários foi capaz de debilitar os tambaquis como observado por meio das alterações hematológicas; sendo este quadro de anemia hipocrômica e trombocitopenia importante para ser utilizado no diagnóstico destas parasitoses. Este estudo foi o primeiro registro da ocorrência de Dolops geayi em tambaquis cultivados na Amazônia.

Changes in hematological and biochemical parameters of tambaqui (Colossoma macropomum ) parasitized by metazoan species.

The aim of this study was to evaluate the impacts of metazoan parasites on hematological and biochemical parameters and relative condition factor of tambaqui (Colossoma macropomum ) farmed in northern Brazil. A total of 32 juvenile fish were captured from a commercial fish farm located in the municipality of Rio Preto da Eva, Amazonas state, Brazil. Parasite prevalence was 100% for Anacanthorus spathulatus, Mymarothecium boegeri and Notozothecium janauachensis, 100% for Neoechinorhynchus buttnerae and 53.13% for Dolops geayi. The greatest mean parasite intensity was found in acantocephalans followed by monogeneans and branchiuran crustaceans. A negative correlation was observed between abundance of N. buttnerae and hematocrit percentage, hemoglobin concentration, total thrombocyte count and glucose and between abundance of the monogenean and glucose concentration. Parasitic infections caused damage in tambaqui in terms of the observed hematological parameters that were characterized by hypochromic anemia and thrombocytopenia, which are important parameters to be used in parasitic diagnosis. This study is the first record of the occurrence of Dolops geayi in farmed tambaqui in the Amazon.

Manipulation of growth of the Amazonian fish tambaqui, Colossoma macropomum (Characiformes: Serrasalmidae): analysis of physiological and zootechnical aspects / Manipulação do crescimento do peixe amazônico tambaqui, Colossoma macropomum (Characiformes: Serrasalmidae): análises fisiológicas e zootécnicas

Colossoma macropomum, known locally as tambaqui, is the native fish most farmed in Brazil, however, technological advances are needed to reach efficient production rates. Modulating growth factors, such as growth hormone, may be associated with improved growth rate and feed efficiency. The use of exogenous hormone for fish rearing is prohibited in Brazil, yet the experimental use of bovine hormone can be useful in research aimed at understanding how to stimulate endogenous growth hormones in fish. Therefore, the present study had the strict objective of understanding the effects of growth hormone on the physiology and zootechnical parameters of C. macropomum under experimental conditions. The animals were intraperitoneally injected every fifteen days with 1, 10 and 100 g g-1 of bGH using 0.9% NaCl saline as diluent. The 10 and 100 g g-1 bolus had a positive effect on the performance indexes of C. macropomum: weight gain (g), growing length (cm), daily weight gain (g), feed conversion and specific growth rate (% per day). The bGH promoted a greater increase in length than in mass, which caused a reduction in condition factor of the individuals receiving a bolus of 10 and 100 g g-1. Furthermore, bGH caused no changes in glucose levels, cortisol, hematological parameters, plasma levels of Na+ and K+, and activity of gills H+-ATPase and Na+, K+-ATPase, at least during the experimental period considered in the present study.

O tambaqui, Colossoma macropomum, é o peixe nativo mais cultivado no Brasil. No entanto, avanços tecnológicos são necessários para incrementar as taxas de produção. Fatores de crescimento moduladores, como o hormônio do crescimento, podem estar associados a uma melhoria na taxa de crescimento e eficiência alimentar. O uso de hormônio exógeno para a criação de peixes é proibido por lei no Brasil, porém, o uso experimental do hormônio bovino pode ser útil em pesquisas que visam determinar mecanismos de estímulo dos hormônios de crescimento endógenos em peixes. Portanto, o presente estudo teve como objetivo estrito a compreensão dos efeitos do hormônio de crescimento sobre a fisiologia e os parâmetros zootécnicos de C. macropomum em condições experimentais . Os animais foram injetados intraperitonealmente a cada quinze dias com 1, 10 e 100 g g-1 de bGH, utilizando solução salina a 0,9% de NaCl como diluente. Observou-se que as concentrações 10 e 100 g g-1 tiveram um efeito positivo sobre os índices de desempenho de C. macropomum em ganho de massa (g), crescimento em comprimento (cm), ganho de massa diário (g), conversão alimentar e taxa de crescimento específico (% por dia). O bGH promoveu ganho maior em comprimento do que em massa, o que causou diminuição do fator de condição nos indivíduos que receberam 10 e 100 g g-1. Além disso, o bGH não causou alterações nos níveis de glicose, cortisol, parâmetros hematológicos, níveis plasmáticos de Na+ e K+ e na atividade de H+-ATPase e Na+, K+-ATPase nas brânquias durante o período experimental.