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The juveniles of Macrobrachium rosenbergii (0.060 ± 0.004 g and 2.018 ± 0.071 cm) were fed during 60 days with diets containing different doses of Lippia alba essential oil (EOLA) (0.0—control, 1.0, or 2.0 ml EOLA/kg diet; in triplicate, with 20 prawns/replicate). After the experimental period, were verified the survival, growth parameters and the antioxidant enzymes, lipid peroxidation and Na/K‐ATPase activities in hepatopancreas and gills, respectively, of the animals. There were no significant differences on survival, growth parameters and gill Na/K‐ATPase activity. However, glutathione peroxidase and glutathione‐S‐transferase activities were lower in hepatopancreas of prawns fed with 1.0 ml EOLA/kg diet compared to the control. Moreover, the activity of the enzyme superoxide dismutase was higher in hepatopancreas of prawns that received 2.0 ml EOLA/kg diet compared to those fed with 1.0 ml EOLA/kg diet. Although the addition of both EOLA doses has not improved the survival and growth parameters of M. rosenbergii, these doses contributed to decrease lipid peroxidation. Additionally, the dose of 2.0 ml EOLA/kg diet contributed for increasing the antioxidant enzymes activities in the hepatopancreas, improving antioxidant status, and therefore, it can be recommended as diet supplementation for M. rosenbergii.

The contents of ions and organic compounds in freshwaters vary to a great extent. In turn, the body fluids of freshwater fish are hyperosmotic in relation to their habitat, making osmoregulatory mechanisms a key feature of their general well-being and survival. This chapter discusses the structure and function of osmoregulatory organs—epithelia, gills, gastrointestinal tract, kidney, and urinary bladder—of Neotropical fish, addressing effects of hypoxia, pH, water hardness, and dissolved organic matter on osmoregulation. Even though a consistent core of knowledge on the subject is recognizable, studies regarding osmoregulatory mechanisms in Neotropical fish remain necessary, given the scarcity of reports on species dwelling in alkaline waters and hard waters, independently of pH range. Studies on the ionic dynamics of the gastrointestinal tract of feral fish can add to the knowledge on osmoregulation of freshwater fish, especially in ion-poor waters, an environment in which it is assumed that a considerable part of the ion uptake derives from ingested food.

Among all organotin compounds , triphenyltin hydroxide (TPhTH) is widely used as fungicide and moluscicide in Brazil. However, the effects of TPhTH on the biochemical parameters of non-target organisms, such as fish, are little known. The aim of the present study is to assess the possible toxic effects of different concentrations of waterborne TPhTH on silver catfish belonging to species Rhamdia quelen. The fish were exposed to two different concentrations of TPhTH (1.08 and 1.70 µg/L as Sn) for 15 days and then compared to the control group (triplicate, n = 3). The antioxidant profile (catalase (CAT) and the glutathione S-transferase (GST)) and the oxidative stress parameters (TBARS-thiobarbituric acid-reactive substances and protein carbonyl (PC)) were set after the exposure to TPhTH. The TBARS level and the PC content increased in several organs of the Rhamdia quelen (brain, liver, muscle and gills) under the two concentrations of TPhTH in comparison to the control group. The CAT activity in the liver and gills has enhanced in all tested TPhTH concentrations. The GST activity increased in the brain, liver and muscle tissues under all the TPhTH concentrations. The significant changes in the biomarkers indicated that the investigated pesticide could have harmful effect on fish, in the field. However, these biomarkers were measured after the fish received doses lower than the recommended for use in agriculture.

The aim of this study was to investigate the anesthetic effect of the essential oils (EOs) of Lippia alba (citral and linalool chemotypes, EOLA-C and EOLA-L respectively) and Lippia origanoides (chemotype carvacrol, EOLO-Ca) on tambaqui (Colossoma macropomum) juveniles. The tested concentrations of the L. alba and L. origanoides EOs were: 25, 50, 100, and 200 μL/L. Time required to reach the stages of sedation, deep anesthesia, and recovery, as well as ventilatory frequency, were monitored for each EO concentration. The results indicated that all EO concentrations tested induced tambaqui anesthesia and that the ventilatory frequency was altered by the EOs. L. origanoides showed the shortest time to induce sedation and anesthesia, but had long recovery times. L. alba EO (citral 100 μL/L and linalool 200 μL/L) can be used to efficiently anaesthetise tambaqui juveniles with an efficient recovery. RESUMO: O objetivo deste estudo foi investigar o efeito anestésico dos óleos essenciais (EOs) de Lippia alba (quimiotipos citral e linalool - EOLA-C e EOLA-L respectivamente) e Lippia origanoides (quimiotipo carvacrol- EOLO-Ca) em juvenis de tambaqui (Colossoma macropomum). As concentrações testadas dos EOs de L. alba e L. origanoides foram: 25, 50, 100 ou 200 μL/L. O tempo para atingir os estágios de sedação, anestesia profunda, recuperação e frequência ventilatória foram monitorados em cada concentração de EO. Os resultados indicam que todas as concentrações testadas dos OE induziram a anestesia do tambaqui e que a frequência ventilatória foi alterada pelos EOs. L. origanoides mostrou o tempo mais rápido para induzir sedação e anestesia, mas os tempos de recuperação foram longos. EO L. alba (citral 100 μL/L e linalol 200 μL/L) pode ser utilizado como anestésico em juvenis de tambaqui, pela eficiência em anestesiar e recuperar.

This study investigated the efficacy of essential oils of Lippia alba (EOLA) citral chemotype and Lippia origanoides (EOLO) and their effects on ventilatory frequency (VF) of silver catfish, Rhamdia quelen. Fish were exposed to 50-300 μL L−1 of EOLA and 25-300 μL L−1 of EOLO to determine induction times to sedation, anesthesia, and recovery. Moreover, VF was determined in fish exposed to 5 or 10 μL L−1 of EOLA and of EOLO for 8 h. The increasing concentration of essential oils proportionally decreased sedation and anesthesia induction times. The highest EOLA concentration increased VF of fish from the control group at 1 h of exposure, but VF decreased at both EOLO concentrations after 2 h. The EOLA citral chemotype and EOLO were effective sedatives and anesthetics for silver catfish. However, EOLO was the most suitable sedative for additional studies regarding fish transport as it reduced VF and did not induce VF increase in silver catfish. The EOLA citral chemotype and EOLO are effective sedatives and anesthetics for silver catfish. Moreover, the EOLO is recommended for transport of silver catfish, because it maintains the ventilatory frequency constant, avoiding a possible metabolic stress.

The effects of the methanolic extract of Condalia buxifolia (MECB) were investigated in silver catfish ( Rhamdia quelen ) transported for 6 h in plastic bags at 0, 5 or 10 μL/L MECB. Prior to transport, the fish were sedated with 10 μL/L MECB for 5 min, except the control group. At the end of transport, dissolved oxygen, alkalinity, pH, temperature, and un-ionized ammonia levels in the transport water were not different between groups, but the control group presented the highest total ammonia levels. Net Na + , Cl − and K + effluxes were highest in fish from the control group compared to those transported with MECB. Pv O 2 , Pv CO 2 and HCO 3 − were higher after transport in fish transported with 5 μL/L MECB, but no significant difference between groups was found regarding blood pH and plasma cortisol levels. The metabolic parameters (glycogen, lactate, total amino acid, total ammonia and total protein) were lower or no significant difference was found in fish transported with MECB. There was no difference between treatments on the ecto-nucleoside triphosphate diphosphohydrolase, 5′-nucleotidase and acetylcholinesterase activities in the whole brain. The activity of all analyzed antioxidants increased in the gills and superoxide dismutase and glutathione-S-transferase also increased in the other analyzed tissues. In addition, lipoperoxidation (measured by thiobarbituric acid-reactive substances) and carbonylation of proteins (measured by protein carbonyl) decreased in most analyzed tissues, indicating lower ROS production. In conclusion, the use of MECB for the transport of silver catfish is advisable because MECB improves antioxidant defenses in several tissues and was effective in reducing waterborne total ammonia levels and ion loss.

The influence of stocking density and food deprivation on energy metabolism, stress processes and the pituitary endocrine system of silver catfish (Rhamdia quelen) was investigated after a period of 14 days, in which plasmatic and hepatic parameters and the mRNA expression of prolactin (PRL), growth hormone (GH) and somatolactin (SL) were assessed. The fish were subjected to four experimental conditions: (1) fed under high stocking density (32 kg/m3, HSD); (2) fed under mean stocking density (16 kg/m3, MSD); (3) fed under low stocking density (8 kg/m3, LSD); and (4) food-deprived under low stocking density (8 kg/m3, LSD-FD). After 14 days, plasma and liver samples were obtained to analyze the metabolite levels and enzymatic activities related to metabolism, and pituitary glands were obtained to analyze hormone expression (PRL, GH and SL). Liver weight and the hepato-somatic index (HSI) revealed that specimens maintained at HSD and/or MSD had higher hepatic stores, which were observed in the triglyceride and glycogen levels in this tissue, than animals submitted to the LSD and LSD-FD groups. Triglyceride levels in the plasma and liver revealed the consumption of fatty acid reserves in the fasting group. Enzymatic activities, such as glutamate dehydrogenase (GDH), phosphorylase (GPase), pyruvate kinase (PK), aspartate transaminase (AST) and glycerol-3-phosphate dehydrogenase (G3PDH), indicated an increase in gluconeogenic pathways in the HSD group and an increase in glycolitic metabolism in the LSD groups. The expression of PRL was not affected by stocking density and/or food deprivation and GH decreased with increased density and increased in fasting conditions. A negative effect of density and fasting was observed on the expression of SL. Overall, the data suggested that juvenile silver catfish reared at stocking densities of 16 to 32 kg/m3 were better maintained than those maintained at the lowest density., We would like to thank the financial support by CAPES (Ministry of Education of Brazil, Brazil) through the program PDSE-CAPES (Programa Institutional de Bolsas de Doutorado Sanduíche no Exterior — Process: 1585/12-6) to C.M., the CNPq research fellowships to V.L.L. and B.B., the project AGL2010-14876 (Ministry of Science and Education, Spain) to J.M.M. and the program FPU (Formación de Profesorado Universitario — Ref. AP2008-01194) (Ministry of Science and Education, Spain) to JAM-S.

The Lophiosilurus alexandri is an important species to the Brazilian aquaculture. Moreover, the studies about the effects of essential oil (EO) of Aloysia triphylla in fish are almost absent. Thus, the aims of this study were: 1) to investigate the anesthetic effect of the EO of A. triphylla and, 2) if this EO improves the physiological parameters of L. alexandri after transport. The experiment was divided in three steps: 1) the anesthesia induction with the EO of A. triphylla (in mu L/L): 25, 50, 100, 200, or 400; 2) transport for 4 h with the fish separated into three groups: control and two EO concentrations of A. triphylla (in mu L/L): 12.5 and 25; and, 3) determination of the ventilatory frequency (VF) in fish exposed to this EO at 12.5 or 25 mu L/L. The results showed that the increasing concentration of A. triphylla EO proportionally decreased the time required for sedation and anesthesia induction and increased recovery time. After transport, some alterations in water and blood parameters were observed independently of the treatment and lower VF was found in fish exposed to 25 mu L/L of A. triphylla EO. Taken together these results indicated that this EO decreased fish metabolism. Finally, A. triphylla EO was an efficient anesthetic in L. alexandri, mainly at 400 mu L/L and, both EO concentrations used in the transport contributed to improve water and physiological parameters. Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) Fundacao de Amparo a Pesquisa do Estado de Minas Gerais (FAPEMIG) CAPES-Brazil CNPq [246702/2012-5]

The transport of live fish is a routine practice in aquaculture and constitutes a considerable source of stress to the animals. The addition of anesthetic to the water used for fish transport can prevent or mitigate the deleterious effects of transport stress. This study investigated the effects of the addition of eugenol (EUG) (1.5 or 3.0 mu L L-1) and essential oil of Lippia alba (EOL) (10 or 20 mu L L-1) on metabolic parameters (glycogen, lactate and total protein levels) in liver and muscle, acetylcholinesterase activity (AChE) in muscle and brain, and the levels of protein carbonyl (PC), thiobarbituric acid reactive substances (TBARS) and nonprotein thiol groups (NPSH) and activity of glutathione-S-transferase in the liver of silver catfish (Rhamdia quelen; Quoy and Gaimard, 1824) transported for four hours in plastic bags (loading density of 169.2 g L-1). The addition of various concentrations of EUG (1.5 or 3.0 mu L L-1) and EOL (10 or 20 mu L L-1) to the transport water is advisable for the transportation of silver catfish, since both concentrations of these substances increased the levels of NPSH antioxidant and decreased the TBARS levels in the liver. In addition, the lower liver levels of glycogen and lactate in these groups and lower AChE activity in the brain (EOL 10 or 20 mu L L-1) compared to the control group indicate that the energetic metabolism and neurotransmission were lower after administration of anesthetics, contributing to the maintenance of homeostasis and sedation status. Fundacao de Amparo a Pesquisa do Estado do Rio Grande do Sul (FAPERGS/PRONEX) [10/0016-8]; Conselho Nacional de Pesquisa e Desenvolvimento Cientifico (CNPq) [470964/2009-0]; Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES); CNPq info:eu-repo/semantics/publishedVersion

This study investigated the anesthetic potential of the essential oil (EO) of Aloysia polystachya in juveniles of dusky grouper (Epinephelus marginatus). Fish were exposed to different concentrations of EO of A. polystachya to evaluate time of induction and recovery from anesthesia. In the second experiment, fish were divided into four groups: control, ethanol and 50 or 300 mu L L-1 EO of A. polystachya, and each group was submitted to induction for 3.5 min and recovery for 5 or 10 min. The blood gases and glucose levels showed alterations as a function of the recovery times, but Na+ and K+ levels did not show any alteration. In conclusion, the EO from leaves of A. polystachya is an effective anesthetic for dusky grouper, because anesthesia was reached within the recommended time at EO concentrations of 300 and 400 mu L L-1. However, most evaluated blood parameters showed compensatory responses due to EO exposure. Fundacao de Amparo a Pesquisa do Estado do Rio Grande do Sul/Programa de Apoio a Nucleos de Excelencia (FAPERGS/PRONEX) [10/0016-8]; Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) [470964/2009-0]; Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior, Brazil (CAPES) info:eu-repo/semantics/publishedVersion

The effects of transporting silver catfish (Rhamdia quelen) for 6 h in plastic bags containing 0 (control), 30 or 40 A mu L/L of essential oil (EO) from Lippia alba leaves were investigated. Prior to transport, the fish in the two experimental groups were sedated with 200 A mu L/L of EO for 3 min. After transport, dissolved oxygen, carbon dioxide, alkalinity, water hardness, pH, temperature and un-ionized ammonia levels in the transport water did not differ significantly among the groups. However, total ammonia nitrogen levels and net Na+, Cl- and K+ effluxes were significantly lower in the groups transported with EO of L. alba than those in the control group. PvO(2), PvCO(2) and HCO3 (-) were higher after transporting fish in 40 A mu L/L of EO of L. alba, but there were no significant differences between groups regarding blood pH or hematocrit. Cortisol levels were significantly higher in fish transported in 30 A mu L/L of EO of L. alba compared to those of the control group. The metabolic parameters (glycogen, lactate, total amino acid, total ammonia and total protein) showed different responses after adding EO to the transport water. In conclusion, while the EO of L. alba is recommended for fish transport in the conditions tested in the present study because it was effective in reducing waterborne total ammonia levels and net ion loss, the higher hepatic oxidative stress in this species with the same EO concentrations reported by a previous study led us to conclude that the 10-20 A mu L/L concentration range of EO and lack of pre-sedation before transport are more effective.

This study investigated the effects of the essential oil (EO) from Lippia alba on biochemical parameters related to oxidative stress in the brain and liver of silver catfish (Rhamdia quelen) after six hours of transport. Fish were transported in plastic bags and divided into three treatments groups: control, 30 µL L- 1 EO from L.alba and 40 µL L-1 EO from L.alba. Prior to transport, the fish were treated with the EO from L. alba (200 µL L -1 for three minutes), except for the control group. Fish transported in bags containing the EO did not have any alterations in acetylcholinesterase, ecto -nucleoside triphosphate diphosphohydrolase and 5'nucleotidase activity in the brain or superoxide dismutase activity in the liver. The hepatic catalase (CAT), glutathione-S-transferase (GST), glutathione peroxidase (GPx), nonprotein thiol and ascorbic acid levels were significantly lower compared to the control group. However, the hepatic thiobarbituric acid- reactive substances, protein oxidation levels and the lipid peroxidation/catalase+glutathione peroxidase (LPO/CAT+GPx) ratio were significantly higher in fish transported with both concentrations of the EO, indicating oxidative stress in the liver. In conclusion, considering the hepatic oxidative stress parameters analyzed in the present experiment, the transport of previously sedated silver catfish in water containing 30 or 40 µL L-1 of EO from L. alba is less effective than the use of lower concentrations. Este estudo investigou os efeitos do óleo essencial (OE) de Lippia alba sobre parâmetros bioquímicos relacionados ao estresse oxidativo em cérebro e fígado de jundiá (Rhamdia quelen), após seis horas de transporte. Os peixes foram transportados em sacos plásticos e divididos em três tratamentos: controle, 30 µL L-1 e 40 µL L-1 de OE de L.alba. Antes do transporte, os peixes foram tratados com o OE de L. alba (200 µL L-1 por três minutos), exceto para o grupo controle. Os peixes transportados em sacos contendo o OE não tiveram alterações na atividade da acetilcolinesterase (AChE), ecto-nucleosídeo trifosfato difosfohidrolase (NTPDase) e 5'nucleotidase, em cérebro ou superóxido dismutase (SOD) no fígado. O tiol não proteico (NPSH), os níveis de ácido ascórbico, catalase (CAT), glutationa-S-transferase (GST) e glutationa-peroxidase (GPx) hepáticos, foram significativamente mais baixos em comparação com o grupo controle. No entanto, as substâncias reativas ao ácido tiobarbitúrico (TBARS), os níveis de oxidação proteica e a taxa de peroxidação lipídica/catalase+glutationa peroxidase (LPO/ CAT+GPx) foram significativamente maiores nos peixes transportados com ambas as concentrações de OE, indicando estresse oxidativo no fígado. Em conclusão, considerando os parâmetros de estresse oxidativo do fígado analisados no presente experimento, o transporte de jundiás previamente sedados em água contendo 30 ou 40 µL L-1 de OE de L.alba é menos efetivo que utilizando concentrações menores.