Joseânia Salbego

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 β-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.

Acetylcholinesterase enzyme activity in carp brain and muscle after acute exposure to diafuran

Exposicao a inseticidas em concentracoes elevadas no ambiente podem ocasionar efeitos adversos subletais em organismos aquaticos. Alevinos de carpa hungara (Cyprinus carpio, Linnaeus, 1758), carpa capim (Ctenopharyngodon idella, Valenciennes, 1844) e carpa cabeca grande (Aristichthys nobilis, Richardson, 1845) foram expostos ao diafuran, um inseticida utilizado na cultura do arroz no sul do Brasil. O objetivo deste estudo foi verificar a relacao entre concentracao letal mediana (CL50) do diafuran e a atividade da enzima acetilcolinesterase (AChE) em cerebro e musculo dessas especies, como um possivel biomarcador inicial da exposicao a este inseticida. A CL50 foi determinada com peixes expostos a concentracoes de diafuran em 96 h: carpa hungara: controle; 0,5; 1,0; 1,5; 2,0; 2,5 e 3,0 mg L-1; carpa capim: controle; 1,0; 2,0; 3,0 e 3,5 mg L-1 e carpa cabeca grande: controle; 0,5; 1,0; 1,5; 2,0; 3,0 e 4,0 mg L-1, bem como a determinacao da AChE em concentracoes proximas da CL50 para essas especies. Valores de CL50 (concentracoes nominais) foram de 1,81 mg L-1 para carpa hungara, 2,71 mg L-1 para carpa capim e 2,37 mg L-1 para carpa cabeca grande. Todas as carpas expostas ao diafuran estavam letargicas (menores concentracoes) ou imoveis. Diafuran inibiu significativamente a atividade da AChE em cerebro (~38 %) e musculo (~50 %) de todas as especies estudadas. Atividade da AChE em musculo para carpa cabeca grande foi mais alta que cerebro (14,44 contra 5,94 µmol min-1 g proteina-1, respectivamente). Este estudo demonstrou que concentracoes de diafuran utilizadas na cultura do arroz podem afetar o comportamento de Cyprinus carpio, Ctenopharyngodon idella e Aristichthys nobilis, e a atividade da acetilcolinesterase cerebral e muscular pode ser um biomarcador inicial de toxicidade deste inseticida.Exposicao a inseticidas em concentracoes elevadas no ambiente podem ocasionar efeitos adversos subletais em organismos aquaticos. Alevinos de carpa hungara (Cyprinus carpio, Linnaeus, 1758), carpa capim (Ctenopharyngodon idella, Valenciennes, 1844) e carpa cabeca grande (Aristichthys nobilis, Richardson, 1845) foram expostos ao diafuran, um inseticida utilizado na cultura do arroz no sul do Brasil. O objetivo deste estudo foi verificar a relacao entre concentracao letal mediana (CL50) do diafuran e a atividade da enzima acetilcolinesterase (AChE) em cerebro e musculo dessas especies, como um possivel biomarcador inicial da exposicao a este inseticida. A CL50 foi determinada com peixes expostos a concentracoes de diafuran em 96 h: carpa hungara: controle; 0,5; 1,0; 1,5; 2,0; 2,5 e 3,0 mg L-1; carpa capim: controle; 1,0; 2,0; 3,0 e 3,5 mg L-1 e carpa cabeca grande: controle; 0,5; 1,0; 1,5; 2,0; 3,0 e 4,0 mg L-1, bem como a determinacao da AChE em concentracoes proximas da CL50 para essas especies. Valores de CL50 (concentracoes nominais) foram de 1,81 mg L-1 para carpa hungara, 2,71 mg L-1 para carpa capim e 2,37 mg L-1 para carpa cabeca grande. Todas as carpas expostas ao diafuran estavam letargicas (menores concentracoes) ou imoveis. Diafuran inibiu significativamente a atividade da AChE em cerebro (~38 %) e musculo (~50 %) de todas as especies estudadas. Atividade da AChE em musculo para carpa cabeca grande foi mais alta que cerebro (14,44 contra 5,94 µmol min-1 g proteina-1, respectivamente). Este estudo demonstrou que concentracoes de diafuran utilizadas na cultura do arroz podem afetar o comportamento de Cyprinus carpio, Ctenopharyngodon idella e Aristichthys nobilis, e a atividade da acetilcolinesterase cerebral e muscular pode ser um biomarcador inicial de toxicidade deste inseticida.

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.

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.

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

No presente estudo foi avaliada a eficacia da anestesia e possiveis efeitos dos oleos essenciais (EOs) de Cunila galioides (EOC) e Origanum majorana (EOO) sobre a taxa ventilatoria (VR) e regulacao ionica em Rhamdia quelen. Nas avaliacoes de anestesia, as concentracoes 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 inducao as fases de sedacao e de anestesia, assim como recuperacao, foram mensurados. Um segundo ensaio empregou concentracoes mais baixas de ambos EOs: 10, 25, 50 e 100 μL L-1 a fim de verificar a VR e o fluxo liquido corporal dos ions Na+, K+ and Cl- . A sedacao foi alcancada para ambos os oleos em 100 μL L-1, e a anestesia em concentracoes ≥ 200 µL L-1. Nao houve diferenca significativa entre o controle e grupos tratados com EOs em relacao a VR, mas todos os peixes submetidos a 100 µL L-1 do EOC morreram dentro de 2 h de exposicao. No geral, a perda ionica declinou na presenca dos EOs. O EOC em 200 - 300 μL L-1 e o EOO em 400 - 500 μL L-1 apresentam potencial para anestesia rapida em R. quelen.

Biochemical parameters of silver catfish (Rhamdia quelen) after transport with eugenol or essential oil of Lippia alba added to the water

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 µL L-1) and essential oil of Lippia alba (EOL) (10 or 20 µL 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 µL L-1) and EOL (10 or 20 µL 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 µL 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.

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.

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.

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.

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)