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)

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.

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.

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.

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)

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)

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.

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.

Hematological and immune changes in Piaractus mesopotamicus in the sepsis induced by Aeromonas hydrophila.

The pathogenesis of sepsis involves complex systems and multiple interrelationships between the host and pathogen producing high mortality rates in various animal species. In this study, hematological disturbances, innate immunity and survival during the septic process in Piaractus mesopotamicus inoculated with Aeromonas hydrophila were studied. For this aim, fish blood samples were taken from control and infected groups 1, 3, 6, and 9 h post-inoculation (HPI). Leukogram showed reduction in the number of leukocytes and thrombocytes, followed by cessation of leukocyte chemotaxis 6 HPI and severe morphological changes in leukocytes and erythrocytes. At 3 HPI production of reactive oxygen species increased and at 6 HPI decreased. There was no change in serum lysozyme concentration and lytic activity of the complement system, despite the progressive increase in serum lytic activity and bacterial agglutination. Finally, the changes in clinical signs due to aeromonosis and increasing septicemia resulted in a reduction in survival to 57.14% after 36 HPI. It was possible concluded that these hematological and immune are crucial event in the worsening of sepsis in P. mesopotamicus, and these findings are utility for diagnosing and understanding the pathophysiology sepsis in pacu induced by A. hydrophila.

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.

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.

Distinct ß-glucan molecules modulates differently the circulating cortisol levels and innate immune responses in matrinxã (Brycon amazonicus).

This study investigated the effects of two ß-glucan molecules with different purities and isolated by different biotechnological processes on the immune response of matrinxã (Brycon amazonicus) prior and after challenge with Aeromonas hydrophila. In this sense, we evaluated serum cortisol and plasma glucose levels, the number of leukocytes (lymphocytes, neutrophils and monocytes), as well as the respiratory activity of leukocytes prior to, 6 and 24 h post infection (hpi). During 15 days, fish were fed with diets containing 0.1% of two ß-glucans (ß-G 1 and ß-G 2, with 71 and 62% of purity, respectively) and then submitted to challenge. Results were compared with a positive control group fed with a ß-glucan-free diet. A negative control group, also fed with ß-glucan-free diet but inoculated with PBS, was established to evaluate the effect of handling during injection. Our results showed that different ß-glucans affected differently the biological responses of matrinxã. The ßG 2 modulated the cortisol profile prior to and after the acute infection with A. hydrophila, and increased the mobilization and activity of leukocytes. The infection promoted lymphopenia at 6 hpi and both ß-glucans increased the circulating lymphocyte population 24 hpi. Moreover, the ß-G 2 prevented the infection-induced neutrophilia at 6 and 24 hpi. Finally, the ß-G 2 caused a marked increase in the circulating monocytes prior to infection, and a reduction at 6 hpi that was reversed at 24 hpi. In summary, our study demonstrates that ß-G 2 was more efficient on the induction of the cell-mediate immunity in matrinxã.

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.

Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus).

The pacu is one of the most important species for Brazilian fish farming and is considered emerging in the global aquaculture. Despite its importance, no effective tool for evaluation of the adaptive immune response of this species has been developed. Therefore, this study aimed the development and standardization of indirect ELISA for the measurement of pacu antigen-specific antibodies using polyclonal rabbit anti-pacu IgM used as detector antibody. For this purpose was isolated and purificated of pacu IgM using mannose-binding protein affinity chromatography and produced specific polyclonal antibodies against heavy and light chains pacu IgM, that showed a molecular weight of 72 kDa and 26 kDa, respectively. Polyclonal antibodies obtained demonstrated specificity with heavy and light Ig chains of pacu serum in western blotting. These polyclonal antibodies allowed the development of an indirect ELISA assay of high sensitivity and specificity for the detection and quantification of pacu IgM antibodies immunized with bovine IgG. In conclusion, this approach has great potential to improve the monitoring of vaccine-induced immune responses and help develop immunodiagnostic and epidemiological studies of infectious diseases in pacu systems.

Glucan-MOS® improved growth and innate immunity in pacu stressed and experimentally infected with Aeromonas hydrophila.

We tested the efficacy of a commercial product (Glucan-MOS®) derived from yeast Saccharomyces cerevisiae, containing two combined products, ß-1,3-1,6 glucans and mannans on the growth, feed efficiency, stress and innate immune responses of juvenile pacu (Piaractus mesopotamicus) after a stressful handling and bacterial inoculation. For this, we evaluated the serum cortisol and plasma glucose levels, the respiratory activity of leukocytes, the serum lysozyme levels, as well as the number of circulating erythrocytes and leukocytes of fish fed during 30 days with diets containing increased levels of Glucan-MOS (0.0, 0.1, 0.2, 0.4 and 0.8%). The supplementation of 0.1% improved weight gain, feed conversion and the protein efficiency ratio compared to a control diet. The 0.2 and 0.4% Glucan-MOS® diets were sufficient to increase the respiratory burst of leukocytes and lysozyme activity, the number of thrombocytes, neutrophils and monocytes in the blood after a stressful handling and bacterial challenge, and minimized stress response as shown by decreased cortisol and glucose levels when compared to the control. The results of this work reinforce the benefits of the adoption of feeding strategies including combination of both ß-1,3-1,6 glucans and mannans as a dietary supplement in periods prior to intensive management. The 30-day period was sufficient to stimulate growth performance, improve nutrient utilization, minimize stress response and modulate innate immunity responses.

Erythrocyte Nuclear Abnormalities in Astyanax lacustris in Response to Landscape Characteristics in Two Neotropical Streams.

The objective of this research was to evaluate land use and occupation in the landscape surrounding two neotropical streams of the Upper Paraná River in Brazil and to investigate if the water from these streams induces erythrocyte nuclear abnormalities in Astyanax lacustris. We observed that the areas surrounding the streams are predominantly used for agricultural purposes (73.46% of the total area), followed by buildings (15.21%). Only 8.78% of the area consisted of forest fragments. We observed higher frequencies of micronuclei in erythrocytes of A. lacustris exposed to water from impacted sites (IMP) compared with less impacted sites and the negative control (NC) (p < 0.05). Analysis of nuclear alterations in A. lacrustis erythrocytes showed significant differences in the frequency of notched nuclei and nuclear pyknosis only between the NC and the IMP sites (p < 0.05). Our results indicate that water from the most impacted sites of the microbasin can induce mutagenic and genotoxic effects in A. lacustris.

Hepatotoxicity and metabolic effects of cellular extract of cyanobacterium Radiocystis fernandoi containing microcystins RR and YR on neotropical fish (Hoplias malabaricus).

The toxicological effect of cellular extract of cyanobacterium Radiocystis fernandoi strain R28 containing RR and YR microcystins was analyzed in the fish Hoplias malabaricus with emphasis on the liver structure and energetic metabolism, after short-term exposure. Fish were intraperitoneally (i.p.) injected with 100 µg of equivalent MC-LR kg-1 body mass containing in the cellular extract of R. fernandoi strain R28. Twelve and 96 h post-injection, the plasma, liver and white muscle were sampled for biochemical analyses and liver was also sampled for morphological analyses. After i.p. injection, the activity of acid phosphatase (ACP), alanine aminotransferase (ALT) and direct bilirubin increased in the plasma, while ALT and aspartate aminotransferase (AST) decreased in the liver. Glucose, lactate and pyruvate increased while protein decreased in the plasma; glycogen, pyruvate and lactate decreased in the liver; and glycogen and glucose increased in the muscle. Ammonia increased in the plasma, liver and muscle. The hepatocyte cell shape changed from polyhedral to round after cellular extract injection; there was loss of biliary canaliculus organization, but the biliary duct morphology was conserved in the liver parenchyma. In conclusion, microcystins present in the cellular extract of R. fernandoi strain R28 affect the liver structure of H. malabaricus, but the liver was able to continuously produce energy by adjusting its intermediate metabolism; glycogenolysis and gluconeogenesis maintained glucose homeostasis and energy supply.