Acute Silver Catfish (Rhamdia quelen) Exposure to Chlorantraniliprole Insecticide.

The aim of the current study is to investigate whether silver catfish (Rhamdia quelen) individuals exposed to commercial formulation of the chlorantraniliprole insecticide used in rice crops present changes in biochemical parameters. Fifty-four (54) silver catfish individuals were distributed in six units per tank (n = 6/repetition; triplicate/treatment) and subjected to the following treatments: T1-control, without insecticide; T2 (0.02 µg/L of insecticide) and T3 (0.20 µg/L of insecticide). Exposure time lasted 24 or 96 h, and it was followed by 96 h recovery in pesticide-free water. Results have indicated biochemical changes in cortisol, glucose, lactate and plasma protein levels, as well as few ionic changes in animals' gills during the exposure and recovery periods. Chlorantraniliprole incidence in water resulted in some biochemical changes in silver catfish specimens' plasma and gills throughout the acute exposure protocol (sub-lethal dose). Thus, chlorantraniliprole insecticide has caused osmoregulatory and/or biochemical imbalance in the investigated species under the herein adopted laboratory conditions; these changes did not get back to normal levels even after specimens were left to recover for 96 h in clean water.

Toxicological response of silver catfish (Rhamdia quelen) after acute exposure to a commercial insecticide containing thiamethoxam.

This study assessed the hematological, enzymatic and osmoregulatory responses of silver catfish (Rhamdia quelen) exposed to sublethal concentrations (1.125 and 3.750 µg/L) of a commercial thiamethoxam-containing insecticide used on rice crops. Groups of 6 fish per tank (in triplicate, n = 3, total 54 fish) were exposed for up to 96 h to different concentrations of the compound. After this period, fish were placed in clean water for 48 h. Two fish from each tank (6 per treatment) that had been exposed to the insecticide for 24 h were anesthetized with eugenol and blood was collected to evaluate hematological and biochemical parameters. Blood, liver and muscle were collected for determination of metabolic parameters, plasma cortisol, Cl-, Na+ and K+ levels and H+-ATPase and Na+/K+-ATPase activity in the gill. H+-ATPase activity was higher in fish exposed to 1.125 µg/L insecticide at 24 h compared to control (0.0 µg/L). Differences in cortisol levels were evidenced throughout the experimental period. These results indicated that exposure to the insecticide changed the hematological, biochemical and metabolic profile of the animals, suggesting concern about environmental safety. Therefore, we discourage the use of this pesticide in areas that come into contact with water bodies inhabited by fish.

The survival and hepatic and muscle glucose and lactate levels of Rhamdia quelen inoculated with Aeromonas hydrophila and treated with terpinen-4-ol, carvacrol or thymol.

The aim of this study was to evaluate the resistance of Rhamdia quelen (silver catfish) to Aeromonas hydrophila infection after treatment with pure and nanoencapsulated forms either terpinen-4-ol, thymol, or carvacrol and the effects of these treatments on fish metabolic responses. After A. hydrophila inoculation, fish were treated with 30 min daily baths for 6 consecutive days with terpinen-4-ol, thymol, or carvacrol in their pure or nanoencapsulated forms at concentrations of 5, 10, 15 or 25 mg L-1. A positive control group, negative control group and saline group were also included. Survival was evaluated at the end of treatment for six consecutive days. Muscle and liver were collected to determine glucose and lactate levels. The fish treated with the nanoencapsulated form of the compounds had a high survival rate, similar to saline group and negative control groups. The carvacrol, thymol and terpinen-4-ol nanoencapsulated forms improved survival of silver catfish infected with A. hydrophila. Muscle and liver glucose and lactate levels are not indicated as biomarkers because they did not present any correlation between the metabolic state of the fish and the bacterial infection.

Gill bioenergetics dysfunction and oxidative damage induced by thiamethoxam exposure as relevant toxicological mechanisms in freshwater silver catfish Rhamdia quelen.

Thiamethoxam is a neonicotinoid pesticide utilized on a worldwide scale, it has been reported in freshwater ecosystems, and detected in fishery products. Nevertheless, there is a lack of information about thiamethoxam sublethal effects on the gills of freshwater fish, principally linked to energetic metabolism. In this context, creatine kinase (CK) is an enzyme of the phosphoryl transfer network that provides a temporal and spatial energy buffer to maintain cellular energy homeostasis in tissues with high energy requirements, such as gills. Based on this evidence, the aim of this study was to evaluate whether exposure to thiamethoxam impairs the cytosolic and mitochondrial CK activities in gills of Rhamdia quelen, and the involvement of oxidative stress in the energetic imbalance. Branchial CK (cytosolic and mitochondrial) activity and sodium­potassium pump (Na+, K+-ATPase) were inhibited, and adenosine triphosphate (ATP) levels decreased after 96 h exposure to 1.125 and 3.75 µg/L thiamethoxam compared to the control group. Moreover, levels of branchial thiobarbituric acid reactive substances (TBARS) and protein carbonylation increased at 3.75 µg/L thiamethoxam after 96 h of exposure compared to the control group, while the non-protein thiol (NPSH) content did not differ between groups. It is important to emphasize that all evaluated parameters did not recover after 48 h in clean water. To summarize, the data presented here clearly demonstrated that thiamethoxan exposure severely impairs cytosolic and mitochondrial CK activities, a key enzyme for gill energy buffering to maintain cellular energy homeostasis, and this effect appears to be mediated by oxidation of lipid and protein molecules, which consequently thereby induces oxidative stress.

Thiamethoxam induced hepatic energy changes in silver catfish via impairment of the phosphoryl transfer network pathway: Toxicological effects on energetics homeostasis.

Precise coupling of spatially separated intracellular adenosine triphosphate (ATP)-producing and ATP-consuming processes exerts a pivotal role in bioenergetic homeostasis of living organisms, and the phosphotransfer network pathway, catalyzed by adenylate kinase (AK) and pyruvate kinase (PK), is fundamental in cellular and tissue energetic homeostasis. Measurement of the phosphotransfer network can provide new information for understanding the alterations in hepatic energetic metabolism during exposition to insecticides, such as thiamethoxam. Therefore, the aim of this study was to evaluate whether exposition to thiamethoxam negatively affects the hepatic enzymes of the phosphotransfer network in silver catfish (Rhamdia quelen). Hepatic AK and PK activities were inhibited at 3.75 µg L-1 after 24 h of exposure and at 1.125 and 3.75 µg L-1 after 96 h of exposure compared with the control group. The hepatic ATP levels were decreased following 3.75 µg L-1 thiamethoxam treatment after 24 h of exposure and at 1.125 and 3.75 µg L-1 after 96 h of exposure compared with the control group. The enzymatic activity of the phosphotransfer network and ATP levels did not recover after 48 h of recovery in clean water. Thus, the inhibition of hepatic AK and PK activities by thiamethoxam caused impairment of energy homeostasis in liver tissue, decreasing hepatic ATP availability. Moreover, the absence of a mutual compensatory mechanism between these enzymes directly contributes to ATP depletion and to a severe energetic dysregulation, which may contribute to toxic effects caused by thiamethoxam.

Purinergic signaling as potential target of thiamethoxam-induced neurotoxicity using silver catfish (Rhamdia quelen) as experimental model.

Thiamethoxam is a broad-spectrum pesticide widely used in agricultural practice throughout the world. Worryingly, this pesticide is considered a potential contaminant on the surface and underground water, being a significant risk to aquatic ecosystems and humans. In this sense, we decided to evaluate the activity of enzymes belonging to purinergic system, which is linked with regulation of extracellular nucleotides and nucleosides, as adenosine triphosphate (ATP) and adenosine (Ado) molecules involved in the regulation of immune and inflammatory responses. Such as the neurotoxic effects of thiamethoxam remain poorly understood, the aim of this study was to evaluate whether purinergic signaling may be considered a potential target of thiamethoxam-induced neurotoxicity in silver catfish (Rhamdia quelen). Brain ectonucleoside triphosphate diphosphohydrolase (ATP as substrate) and 5'-nucleotidases activities were inhibited at 3.75 µg L-1 after 24 h of exposure and at 1.125 and 3.75 µg L-1 after 96 h of exposure compared with the control group. On the other hand, brain adenosine deaminase activity was stimulated at 3.75 µg L-1 after 24 h of exposure and at 1.125 and 3.75 µg L-1 after 96 h of exposure compared with the control group. Brain ATP levels increased at 3.75 µg L-1 after 24 h of exposure and at 1.125 and 3.75 µg L-1 after 96 h of exposure compared with the control group, while the Ado levels decreased. The enzymatic activity of the purinergic signaling did not return to control levels after a 48-h recovery period, revealing the potential neurotoxic effects of thiamethoxam. In summary, the brain purinergic signaling may be considered a potential target for thiamethoxam-induced neurotoxicity in silver catfish.

Essential oils from Citrus x aurantium and Citrus x latifolia (Rutaceae) have anesthetic activity and are effective in reducing ion loss in silver catfish ( Rhamdia quelen )

This study investigated the anesthetic effect of the essential oils (EOs) from the peel of Citrus x aurantium (EOCA) and Citrus x latifolia (EOCL) on silver catfish Rhamdia quelen. Fish were exposed to different concentrations of EOCA and EOCL to determine time of anesthesia induction and recovery. Induction of anesthesia was observed in all fish exposed to 400, 600 or 800 µL L−1 EOCA and 300, 400 or 500 µL L−1 EOCL. Another group of fish were exposed for 8 h to 50, 100, or 200 µL L−1 of either EOs. Overall, fish exposed to ethanol and both EOs presented higher ventilatory frequencies (VF) than the control group throughout the 8 h of exposure. Net ion (Na+, K+ and Cl−) effluxes and ammonia excretion were significantly lower in fish exposed to 50, 100 or 200 µL L−1 of either EOs compared to control fish. Mortality was 37% in fish exposed to 200 µL L−1 of either EOs after 8 h. These findings suggest that EOCA and EOCL are useful anesthetics and sedatives for Rhamdia quelen, but their usefulness as alternatives to reduce stress in fish transportation at the lower concentrations tested (50-100 µL L−1) deserves further study.(AU)

O efeito anestésico dos óleos essenciais (OEs) da casca de Citrus x aurantium (OECA) e Citrus x latifolia (OECL) em jundiá Rhamdia quelen foi investigado. Os peixes foram expostos a diferentes concentrações de OECA e OECL para determinar o tempo de indução e recuperação da anestesia. Todos peixes expostos a 400, 600 ou 800 µL L−1 OECA e 300, 400 ou 500 µL L−1 OECL foram anestesiados. Outro grupo de peixes foi exposto aos OEs durante 8 h a 50, 100 ou 200 µL L−1. Peixes expostos ao etanol e aos OEs apresentaram VF maior que o grupo controle durante as 8 h de exposição. Os efluxos líquidos de Na+, K+, Cl− e a excreção de amônia foram significativamente menores nos peixes expostos a 50, 100 ou 200 µL L−1 dos OEs em comparação com o grupo controle. A mortalidade foi de 37% nos peixes expostos a 200 µL L−1 de ambos os OEs após 8 h. Os resultados sugerem que OECA e OECL são anestésicos e sedativos úteis para o jundiá, mas sua utilidade como alternativa para reduzir o estresse no transporte de peixes nas concentrações mais baixas testadas (50-100 µL L−1) necessita de estudos adicionais.(AU)

Anesthesia and sedation of map treefrog (Hypsiboas geographicus) tadpoles with essential oils / Anestesia e sedação de girinos da perereca geográfica Hypsiboas geographicus com óleos essenciais

ABSTRACT: The goal of this study was to investigate the sedative and anesthetic properties of essential oils (EOs) in map treefrog tadpoles (Hypsiboas geographicus) and to determine the sedation and deep anesthesia induction times as well as the recovery time. The tadpoles were exposed to one of the EOs from three plant species: Aniba rosaeodora (EOAR - 25, 50, 100 or 200µL L-1), Lippia origanoides (EOLO - 13, 25, 50, 100 or 200µL L-1), and Lippia alba (either chemotype citral [EOL-C - 25, 50, 100 or 200µL L-1] or linalool [EOL-L - 50, 75, 100 or 200µL L-1]) (n = 8 per replicate). The tadpoles exposed to 25 and 50µL L-1 EOL-C and EOL-L, respectively, were not anesthetized within 30min (the maximum time of observation), and those exposed to 200µL L-1 EOLO did not recover within 30min. Sedation, deep anesthesia and recovery times showed a concentration-dependent relationship for all EOs tested, with the exception of the recovery with EOLO. The results allowed concluding that all investigated EOs can be used to anesthetize tadpoles of H. geographicus, but the use of EOLO must not exceed 100µL L-1.

RESUMO: O objetivo deste estudo foi investigar as propriedades sedativas e anestésicas de óleos essenciais (OEs) em girinos da perereca Hypsiboas geographicus e determinar os tempos de indução à sedação e anestesia profunda, bem como o de recuperação. Os girinos foram expostos a um dos OEs de três espécies de plantas: Aniba rosaeodora (OEAR - 25, 50, 100 ou 200µL L-1), Lippia origanoides (OELO - 13, 25, 50, 100 ou 200µL L-1 ) ou Lippia alba quimiotipos citral (OEL-C - 25, 50, 100 ou 200µL L-1) ou linalol (OEL-L - 50, 75, 100 ou 200µL L-1) (n = 8 cada repetição). Girinos expostos a 25 e 50µL L-1 OEL-C e OEL-L, respectivamente, não foram anestesiados dentro de 30min (tempo máximo de observação) e aqueles expostos a 200µL L-1 OELO não recuperaram dentro de 30min. Os tempos de sedação, anestesia profunda e recuperação apesentaram uma relação concentração-resposta para todos os OEs testadas, exceto a recuperação com OELO. Os resultados permitem concluir que todos os OEs investigados podem ser usados para anestesiar os girinos de H. geographicus, mas o uso de OELO não deve ser superior a 100µL L-1.

Oxidative and biochemical responses in Brycon amazonicus anesthetized and sedated with Myrcia sylvatica (G. Mey.) DC. and Curcuma longa L. essential oils.

OBJECTIVE: To investigate the effects of rapid anesthesia and long-term sedation with the essential oils (EOs) of Myrcia sylvatica (EOMS) and Curcuma longa (EOCL) on biochemical and oxidative parameters in matrinxã. STUDY DESIGN: Prospective, randomized, laboratory experiment. ANIMALS: A total of 72 matrinxã (Brycon amazonicus) adults weighing 404.8 ± 27.9 g were divided into eight groups of nine fish. METHODS: Biochemical and oxidative effects were investigated in plasma and tissues of matrinxã subjected to rapid anesthesia (5 minutes) or long-term sedation (360 minutes, simulating the practice of transport) with EOMS (200 µL L-1 and 10 µL L-1, respectively) and EOCL (500 µL L-1 and 40 µL L-1, respectively). RESULTS: Transport simulation without sedation or anesthesia increased lipid peroxidation levels in the gills and kidney of fish in the control group. Anesthesia and sedation with EOs decreased cortisol concentrations and increased lactate concentrations compared with controls. Lipid peroxidation was lower in the brain, gills, liver and kidney of sedated and anesthetized fish, than in the control group. Anesthesia with EOs increased the activity of superoxide dismutase and glutathione-S-transferase in the brain, and catalase in the liver and gills, compared with controls. Long-term sedation with EOs increased superoxide dismutase, glutathione peroxidase and glutathione reductase activities in the brain, catalase in the liver, glutathione peroxidase and glutathione reductase in the gills and superoxide dismutase in the kidney. In general, nonprotein thiols content and total reactive antioxidant potential of tissues were higher after anesthesia and sedation with EOs compared with the control group. CONCLUSIONS AND CLINICAL RELEVANCE: The concentrations of EOMS and EOCL used were effective at preventing a stress response and excess of reactive oxygen species formation. For these reasons, these substances may be recommended for use in the transportation of fish to improve survival and animal welfare.

Physiological responses of Rhamdia quelen (Siluriformes: Heptapteridae) to anesthesia with essential oils from two different chemotypes of Lippia alba

The aim of this study was to evaluate if Lippia alba has different chemotypes according to the chemical composition of the essential oil (EO) considering collection site, and if the EO may have different effects on blood and plasma parameters in silver catfish, Rhamdia quelen, during and immediately after anesthesia. The citral (EO-C) and linalool (EO-L) chemotypes were identified, and both presented similar anesthetic effects for silver catfish. Fish were exposed to two concentrations of each EO, which induced slow and fast anesthesia (100 and 300 µL L-1, respectively). Blood ions did not change at any time of anesthesia induction and recovery and, therefore, the electrolyte balance was not altered. Blood gases oscillated through all exposure and recovery times, but there was an increase in pO2 after 10 min recovery in fish anesthetized with EO-C. Glucose increased in fish exposed to both EOs when compared with the control group. Overall, exposure to both EOs (except 100 µL L-1 EO-L at most times) reduced plasma cortisol levels compared to the control and/or ethanol groups. However, as plasma creatinine levels in fish anesthetized with EO-C were higher than control fish, the use of EO-L is preferable.(AU)

O objetivo deste estudo foi avaliar se Lippia alba apresenta diferentes quimiotipos de acordo com a composição química do óleo essencial (OE), considerando local de coleta e se o OE causa diferentes efeitos nos parâmetros sanguíneos e plasmáticos em jundiá, Rhamdia quelen, durante e imediatamente após a anestesia. Os quimiotipos citral (OE-C) e linalol (OE-L) foram identificados e ambos apresentaram efeito anestésico semelhante para jundiá. Os peixes foram expostos a duas concentrações de cada OE, que induziram anestesia lenta e rápida (100 e 300 mL L-1, respectivamente). Íons sanguíneos não se alteraram em nenhum tempo e consequentemente, o equilíbrio eletrolítico não foi alterado. Os gases sanguíneos oscilaram durante todo tempo de exposição e recuperação, mas houve aumento na pO2 após 10 min de recuperação em peixes anestesiados com OE-C. Níveis sanguíneos de glicose aumentaram nos peixes expostos a ambos OEs quando comparados com o grupo controle. De um modo geral, a exposição a ambos OEs (exceto 100 µL L-1 OE-L na maioria dos tempos) reduziu o cortisol plasmático comparado aos grupos controle e etanol. No entanto, como os níveis de creatinina plasmática em peixes anestesiados com OE-C foram maiores que nos peixes controle, é preferível o uso do OE-L.(AU)

Essential oils of Cunila galioides and Origanum majorana as anesthetics for Rhamdia quelen: efficacy and effects on ventilation and ionoregulation

This study evaluated anesthetic efficacy and possible effects of the essential oils (EOs) of Cunila galioides (EOC) and Origanum majorana (EOO) on ventilatory rate (VR) and ionoregulation in Rhamdia quelen. In the anesthesia assessments, 50, 100, 200 and 300 μL L-1 EOC and 50, 100, 200, 300, 400 and 500 μL L-1 EOO were tested, and time for induction to sedation and anesthesia stages, as well as recovery, were taken. A second trial employed lower concentrations of both EOs, 10, 25, 50 and 100 μL L-1, in order to verify VR and Na+, K+ and Cl- whole body net fluxes. Sedation was achieved with both oils at 100 µL L-1, and anesthesia at ≥ 200 µL L-1. There was no significant difference between control and EO-treated groups regarding VR, but all fish subjected to 100 µL L-1 EOC died within 2 h of exposure. Overall, ionic loss declined in the presence of the EOs. The EOC at 200 - 300 μL L-1 and EOO at 400 - 500 μL L-1 present the potential to promote fast anesthesia in R. quelen.(AU)

No presente estudo foi avaliada a eficácia da anestesia e possíveis efeitos dos óleos essenciais (EOs) de Cunila galioides (EOC) e Origanum majorana (EOO) sobre a taxa ventilatória (VR) e regulação iônica em Rhamdia quelen. Nas avaliações de anestesia, as concentrações de 50, 100, 200 e 300 μL L-1 EOC e 50, 100, 200, 300, 400 e 500 μL L-1 EOO foram testadas, e os tempos para a indução às fases de sedação e de anestesia, assim como recuperação, foram mensurados. Um segundo ensaio empregou concentrações mais baixas de ambos EOs: 10, 25, 50 e 100 μL L-1 a fim de verificar a VR e o fluxo líquido corporal dos íons Na+, K+ and Cl- . A sedação foi alcançada para ambos os óleos em 100 μL L-1, e a anestesia em concentrações ≥ 200 µL L-1. Não houve diferença significativa entre o controle e grupos tratados com EOs em relação a VR, mas todos os peixes submetidos a 100 µL L-1 do EOC morreram dentro de 2 h de exposição. No geral, a perda iônica declinou na presença dos EOs. O EOC em 200 - 300 μL L-1 e o EOO em 400 - 500 μL L-1 apresentam potencial para anestesia rápida em R. quelen.(AU)

Imazapyr+imazapic herbicide determines acute toxicity in silver catfish Rhamdia quelen.

Imazapyr (IMY) and imazapic (IMI) are imidazolinone herbicides which have been associated in a commercial formulation (Kifix(®)). To date, there are no studies on the toxicity of an IMY+IMI herbicide in fish. This work aimed to assess the acute toxicity (24 and 96 h) of IMY+IMI (0, 0.488 and 4.88 µg/L) towards Rhamdia quelen through hematological, biochemical, immunological, ionoregulatory and enzymatic indexes. Red blood cell count was lower at 4.88 than at 0.488 µg/L (24 and 96 h); mean corpuscular volume was lower than control at both concentrations (24 h) and at 0.488 µg/L (96 h); lymphocytes declined at 4.88 µg/L comparing to control (96 h); and monocytes increased at 4.88 µg/L (96 h) in comparison with the respective control and with 4.88 µg/L at 24h. Aspartate aminotransferase was higher at 0.488 µg/L (96 h) than the respective control and the respective concentration at 24 h; uric acid reduced at 4.88 µg/L comparing with 0.488 µg/L (96 h); and cortisol was lower at 4.88 µg/L compared to 0.488 µg/L and control (96 h). Herbicide exposure lowered plasma bactericidal activity at both concentrations (24 h) and at 0.488 µg/L (96 h); and plasma complement activity declined at 4.88 µg/L comparing with 0.488 µg/L and control (96 h), and was lower at all concentrations at 96 h than at 24 h. Plasma K(+) levels were higher at 4.88µg/L than in the remaining groups (24 and 96h); and Na(+) levels decreased at 4.88 µg/L compared to control (96 h). Na(+)/K(+)-ATPase and H(+)-ATPase activities in gills were lower at 4.88 µg/L comparing with control (24 h) and with the respective concentration at 96 h; and AChE activity in brain was higher at 0.488 and 4.88 µg/L than control (24 h) and the respective concentrations at 96 h, while in muscle it was higher at 0.488 and 4.88 µg/L than control (96 h) and the respective concentrations at 24 h. The present findings demonstrate that, despite IMY+IMI targets the animal-absent AHAS enzyme, such formulation displayed an acute toxic effect upon R. quelen homeostasis by impacting on vital functions such as immune defense, metabolism, ionoregulation and neurotransmission.

Exposure to different glyphosate formulations on the oxidative and histological status of Rhamdia quelen.

Due to the wide use of glyphosate (GLY) in soybean cultivation, their residues in the environment may affect non-target organisms such as fish, developing toxic effects. Despite GLY being widely used in Brazil, there are few studies comparing the effects of commercial formulations in native freshwater fish species. Silver catfish (Rhamdia quelen) were exposed to three different commercial formulations of GLY 48% (Orium(®), Original(®) and Biocarb(®)) at 0.0, 2.5 and 5.0 mg/L for 96 h. The effects in thiobarbituric acid-reactive substances (TBARS), catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST) and histological alterations were analysed in the liver, whereas alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were studied in the plasma. In the liver, TBARS levels increased and CAT decreased in all treatments and herbicides tested in comparison with the control group. The SOD increased at 2.5 mg/L of Orium(®), Original(®) and 5.0 mg/L Orium(®) and Biocarb(®), whereas GST increased at 2.5 mg/L Orium(®) and decreased at 2.5 mg/L Biocarb(®) when compared to the control group. The main histopathological alterations in hepatic tissue were vacuolisation, leucocyte infiltration, degeneration of cytoplasm and melanomacrophage in all GLY treatments. The ALT decreased after exposure to 2.5 mg/L of Biocarb(®) and AST increased at 2.5 mg/L of Orium(®), Original(®) and 5.0 mg/L of Biocarb(®) in comparison with the control group. In summary, the oxidative damage generated by GLY may have caused the increased formation of free radicals that led to the histological alterations observed in hepatocytes.

Pre-sedation and transport of Rhamdia quelen in water containing essential oil of Lippia alba: metabolic and physiological responses.

The effects of transporting silver catfish (Rhamdia quelen) for 6 h in plastic bags containing 0 (control), 30 or 40 µL/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 µL/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. PvO2, PvCO2 and HCO3(-) were higher after transporting fish in 40 µL/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 µL/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 µL/L concentration range of EO and lack of pre-sedation before transport are more effective.

Rhamdia quelen (Quoy & Gaimard, 1824), submitted to a stressful condition: effect of dietary addition of the essential oil of Lippia alba on metabolism, osmoregulation and endocrinology

The aim of this study was to evaluate the effect of the essential oil of Lippia alba (EOLA) as a feed additive on ionoregulatory and metabolic parameters and pituitary hormones expression in silver catfish, Rhamdia quelen , submitted to a stressful condition (stocking density of 10.6 kg m-3 and limited space). Fish were fed with different concentrations of EOLA (0.0 - control, 0.25 and 0.50 mL kg food-1) for 20 days. Metabolic parameters were not affected by the diet, with the exception of alanine aminotransferase, which was higher in the liver of fish fed 0.50 mL EOLA kg food-1. Plasma ions and activity of H+-ATPase did not change, but fish fed 0.25 mL EOLA kg food-1 presented higher Na+/K+-ATPase activity. Somatolactin expression in the pituitary was higher in the fish fed 0.25 mL EOLA kg food -1, but the expression of growth hormone and prolactin did not change. Therefore, dietary EOLA does not exert a protective effect in R. quelen submitted to a stressful situation because it did not alter most measured parameters. The use of 0.25 mL EOLA kg food-1 seems to be more suitable than 0.50 mL EOLA kg food-1 since the latter may be related to liver damage.

O objetivo deste estudo foi avaliar o efeito do óleo essencial de Lippiaalba (OELA) como aditivo em rações na ionoregulação, parâmetros metabólicos e expressão de hormônios hipofisários em jundiás, Rhamdiaquelen, submetidos a uma situação estressante (densidade de estocagem de 10,6 kg m-3 e espaço limitado). Os peixes foram alimentados com diferentes concentrações de OELA (0,0 - controle, 0,25 e 0,50 mL kg de ração-1) durante 20 dias. Parâmetros metabólicos não foram afetados pela dieta, com a exceção da alanina aminotransferase, que foi mais elevada no fígado dos peixes alimentados com 0,50 mL de OELA kg de ração-1. Íons plasmáticos e a atividade da H+-ATPase não apresentaram nenhuma alteração, mas os peixes alimentados com 0,25 mL OELA kg de ração-1 apresentaram maior atividade da Na+/K+-ATPase. A expressão da somatolactina na hipófise de peixes alimentados com 0,25 mL OELA kg de ração-1 aumentou, porém a expressão do hormônio de crescimento e da prolactina não mudou. Portanto, a adição do OELA na ração não tem um efeito protetor em jundiás submetidos a uma situação estressante, pois não influiu na maioria dos parâmetros medidos. O uso de 0,25 mL OELA kg de ração-1 parece ser mais adequado que 0,50 mL OELA kg de ração-1, uma vez que este nível de inclusão pode estar relacionado a danos hepáticos.

Anesthetic activity and bio-guided fractionation of the essential oil of Aloysia gratissima (Gillies & Hook.) Tronc. in silver catfish Rhamdia quelen.

This work aimed to determine the efficacy of the essential oil of A. gratissima as anesthetic for silver catfish, and to perform the bio-guided fractionation of essential oil aiming to isolate compounds responsible for the noted effects. Fish were submitted to anesthesia bath with essential oil, its fractions and isolated compounds to determine time of anesthetic induction and recovery. Eugenol (50 mg L(-1)) was used as positive control. Essential oil of A. gratissima was effective as an anesthetic at concentrations of 300 to 900 mg L(-1). Fish presented involuntary muscle contractions during induction and recovery. The bio-guided fractionation of essential oil furnished E-(-)-pinocamphone, (-)-caryophyllene oxide, (-)-guaiol and (+)-spathulenol. E-(-)-pinocamphone caused the same side effects observed for essential oil. (-)-Caryophyllene oxide, (-)-guaiol and (+)-spathulenol showed only sedative effects at proportional concentrations to those of the constituents in essential oil. (+)-Spathulenol (51.2 mg L(-1)) promoted deep anesthesia without side effects. A higher concentration of (+)-spathulenol, and lower or absent amounts ofE-(-)-pinocamphone could contribute to increase the activity and safety of the essential oil of A. gratissima. (+)-Spathulenol showed potent sedative and anesthetic activities in silver catfish, and could be considered as a viable compound for the development of a new anesthetic.

The essential oil from Lippia alba induces biochemical stress in the silver catfish (Rhamdia quelen) after transportation

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.

Glyphosate on digestive enzymes activity in piava (Leporinus obtusidens) / Glifosato sobre a atividade de enzimas digestivas em piavas (Leporinus obtusidens)

The effects of glyphosate, a nonselective herbicide (1.0 or 5.0mg L-1) on digestive enzymes activity (stomach and intestine) were evaluated in juveniles of piava (Leporinus obtusidens) after 90 days of exposure. The activity of acid protease, trypsin, chymotrypsin and amylase increased with the increase of glyphosate concentration. These results indicate that glyphosate affects digestive enzyme activities in this species, and may be an indicator of poor nutrient availability when fish survive in herbicide-contaminated water.

Os efeitos do glifosato, um herbicida não seletivo (1,0 ou 5,0mg L-1), sobre a atividade de enzimas digestivas (estômago e intestino) foram avaliadas em juvenis de piava (Leporinus obtusidens) após 90 dias de exposição. A atividade da protease ácida, tripsina, quimiotripsina e amilase aumentaram com a elevação da concentração de glifosato. Esses resultados indicam que o glifosato afeta a atividade de enzimas digestivas nesta espécie e pode ser indicador da reduzida disponibilidade de nutrientes, quando peixes sobrevivem em água contaminada com este herbicida.

The effects of ammonia and water hardness on the hormonal, osmoregulatory and metabolic responses of the freshwater silver catfish Rhamdia quelen.

The aim of the present study was to assess the effects of ammonia and water hardness on endocrine, osmoregulatory and metabolic parameters in silver catfish (Rhamdia quelen). The specimens (60-120g) were subjected to six treatments in triplicate, combining three levels of un-ionized ammonia (NH3) (0.020±0.008mg/L [1.17±0.47µM], 0.180±0.020mg/L [10.57±1.17µM] and 0.500±0.007mg/L [29.36±0.41µM]) and two levels of water hardness (normal: 25mgCaCO3/L and high: 120mgCaCO3/L), and sampled after two exposure times (1 and 5 days post-transfer). Plasma cortisol, metabolites, osmolality and ionic values were determined concomitantly with the mRNA expression levels of different adenohypophyseal hormones (growth hormone, GH; prolactin, PRL; and somatolactin, SL). Previously, full-length PRL and SL as well as ß-actin cDNAs from R. quelen were cloned. Exposure to high NH3 levels enhanced plasma cortisol levels in fish held under normal water hardness conditions but not in those kept at the high hardness value. The increase in water hardness did not alter plasma metabolites, whereas it modulated the osmolality and ion changes induced by high NH3 levels. However, this hardness increase did not lead to the decreased GH expression that was observed 5 days after exposure to 0.18mg/L NH3 in fish held at the normal water hardness level, whereas PRL expression was enhanced after one day of exposure under the increased hardness conditions. Additionally, SL expression decreased in specimens exposed for 5 days to 0.18mg/L NH3 and maintained at the high water hardness level. The results showed that increasing water hardness attenuated the hormonal parameters evaluated in R. quelen specimens exposed to high NH3 levels, although plasma metabolism do not appear to suffer major changes.

Exposure to sublethal concentrations of copper changes biochemistry parameters in silver catfish, Rhamdia quelen, Quoy & Gaimard.

The effects of Cu exposure on catalase (CAT) and acetylcholinesterase (AChE) activity, formation of thiobarbituric acid-reactive species (TBARS) and metabolic parameters were evaluated in silver catfish (Rhamdia quelen). The fish were exposed for 45 days to 0, 16 and 29 µg/L Cu. The fish that were exposed to Cu exhibited lower TBARS levels in the muscle and higher TBARS levels in the liver. They also showed lower CAT activity in the liver and lower AChE activity in the brain and muscle. Higher glucose and lactate and lower protein plasma levels were observed in the fish exposed to Cu. The changes in the hepatic metabolic parameters were Cu concentration dependent. In the muscle, lower glycogen and higher lactate levels were observed in the fish exposed to Cu. Alterations in the metabolic parameters showed a preference for the anaerobic pathway of energy production and liver protein catabolism to supply the energy demand.