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The physiological and behavioral responses of Pomacea canaliculata exposed to different pH values (4, 5, 6, 7, 8, 9, and 10) were evaluated. Survival, behavior (avoidance), metabolites (mantle), net ion fluxes (Na+ and K+), and ATPase activity (gills) were the parameters analyzed. The final survival rates were 100% (pH 4-9) and 90% (pH 10), and the groups did not differ significantly. Avoidance behavior was not identified in animals exposed to an extreme pH compared to pH 7. The main changes observed in the metabolites were in those exposed to an alkaline pH. Glucose (pH 9) and total protein (pH 9 and 10) levels increased, and lactate decreased (pH 9 and 10) compared to a neutral pH. There was an increase in Na+ efflux at pH 4, 5, and 8 and an influx at pH 9 and 10. Extreme pH values (4 and 10) also caused an increase in K+ efflux. At pH values outside the neutrality range (pH 7), there was a significant decrease in the activities of Na+/K+-ATPase (4, 5, 6, 9, and 10) and H+-ATPase (pH 4, 5, 9, and 10). Variations in environmental pH did not cause statistically significant mortality or avoidance behavior in P. canaliculata at the analyzed times. However, due to changes in energy metabolism (glucose and lactate, mainly) and ionoregulation, these can be considered sensitive biomarkers of stress in this species.

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Fish are useful animal models in research and have been employed in developing new pharmacological approaches. This study aimed to establish the use of silver catfish (Rhamdia quelen) as an animal model to evaluate antinociceptive activity. Initially, different concentrations of acetic acid (2.5-20%), formalin 1% (1-10 µL), menthol 0.5% (1-10 µL) or vehicle were injected in the lips to establish which concentration of each sample promotes nociceptive-like behavior in various parameters. The effect of morphine (0.5-10 mg/kg) on locomotion parameters was also evaluated for antinociceptive concentration determination. Morphine was administered intramuscularly immediately prior to algogen administration. The inhibition was evaluated with the antagonist naloxone (5 mg/kg), which was administered in the same way. Recording time varied according to the algogen used in each test and locomotor activity was evaluated by ANY-maze® software. Acid acetic at 15%, 10 µL of 1% formalin, and 1 µL of 0.5% menthol were chosen since they promoted nociceptive-like behavior in several parameters. Morphine (5 mg/kg) reversed the algogen-induced nociceptive-like behavior and naloxone inhibited this effect. Therefore, the proposed experimental model demonstrated specificity for nociception, since the reversion of the nociceptive-like behavior for a compound with well-described analgesic activity was observed. This new pharmacological model contributes to evaluating compounds with analgesic potential and developing new analgesic drugs, in addition to being a promising alternative to use with rodents.

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In teleost fish, stress initiates a hormone cascade along the hypothalamus-pituitary-interrenal (HPI) axis to provoke several physiological reactions in order to maintain homeostasis. In aquaculture, a number of factors induce stress in fish, such as handling and transport, and in order to reduce the consequences of this, the use of anesthetics has been an interesting alternative. Essential oil (EO) of Lippia alba is considered to be a good anesthetic; however, its distinct chemotypes have different side effects. Therefore, the present study aimed to investigate, in detail, the expression of genes involved with the HPI axis and the effects of anesthesia with the EOs of two chemotypes of L. alba (citral EO-C and linalool EO-L) on this expression in silver catfish, Rhamdia quelen. Anesthesia with the EO-C is stressful for silver catfish because there was an upregulation of the genes directly related to stress: slc6a2, crh, hsd20b, hspa12a, and hsp90. In this study, it was also possible to observe the importance of the hsd11b2 gene in the response to stress by handling. The use of EO-C as anesthetics for fish is not recommended, but, the use of OE-L is indicated for silver catfish as it does not cause major changes in the HPI axis.

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In aquaculture, nutrition and supplemented diets have been shown to affect broodstock reproductive performance. In this study, we investigated the effects of dietary supplementation with Cymbopogon flexuosus essential oil (CFEO) microcapsules on reproductive-related parameters in silver catfish (Rhamdia quelen) male broodfish. Adult male broodstocks were separated into three groups according to the concentrations of supplemented CFEO (0.0 = control; 1.0 or 3.0 mL per kg of diet). After 20 days under experimental conditions, the animals were euthanized and the gonads were harvested for gonadosomatic index, sperm analysis, oxidative stress, and histopathology; testosterone levels were measured in the plasma; gene expression of prl, smtl, pomca, and pomcb was assessed in the pituitary gland by real-time PCR. The results showed no alterations on reproductive parameters in R. quelen males treated with Cymbopogon flexuosus essential oil compared to the control-diet animals. In conclusion, CFEO microcapsules supplied for 20 days in the concentrations of 1.00 or 3.00 mL per kilogram of diet did not affect the reproduction criteria evaluated in this study in male silver catfish.

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The viability using Lippia alba essential oil as an anesthetic for fish was studied, particularly with respect to physiological effects during recovery. Anesthesia of silver catfish (Rhamdia quelen) using 100 and 300 µL L-1 of two different chemotypes of L. alba essential oil (citral EO-C and linalool EO-L) prevented the increase of plasma cortisol levels caused by handling, but did not avoid alterations in energetic metabolism. Silver catfish did not have increased the levels of thiobarbituric acid reactive species in the kidney and liver during recovery after anesthesia with either EO, avoiding lipid damage. On the other hand, fish anesthetized with EO-C showed higher protein carbonylation levels, superoxide dismutase, catalase, and glutathione S-transferase activities and non-protein thiol group levels in both tissues compared to controls. Our results suggest that both oils show antioxidant capacity, but anesthesia with EO-L does not cause damage to lipids or proteins, only temporary changes, typical of physiological adjustments during recovery from anesthesia. Therefore, EO-L is an effective anesthetic for silver catfish with fewer side effects than EO-C.

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This study evaluated the potential relaxing and/or molluscicidal effects of eugenol and essential oils of Origanum majorana, Ocimum americanum, Hesperozygis ringens, and Piper gaudichaudianum in the gastropod Pomacea canaliculata. Compounds were tested at concentrations of 100, 250, 500, and 750µL L-1 to evaluate the relaxing effects. In the second experiment, animals were exposed to 10, 25, and 50µL L-1 of essential oils of H. ringens and P. gaudichaudianum for a period of 24h for the evaluation of molluscicidal effects. Eugenol and essential oils of O. majorana and O. americanum showed relaxing effects at 250µL L-1, but the essential oils of H. ringens and P. gaudichaudianum did not promote relaxing or molluscicidal effects within the times and concentrations studied. Therefore, only eugenol and the essential oils of O. majorana and O. americanum can be used for relaxation purposes in P. canaliculata.

Este estudo teve como objetivo avaliar os possíveis efeitos relaxantes e/ou moluscicidas do eugenol e óleos essenciais de Origanum majorana, Ocimum americanum, Hesperozygis ringens e Piper gaudichaudianum no gastrópode Pomacea canaliculata. Os compostos foram testados nas concentrações de 100, 250, 500 e 750µL L-1 para avaliar os efeitos relaxantes. Em um segundo experimento, os animais foram expostos a 10, 25 e 50µL L-1 por 24 horas aos óleos essenciais de H. ringens e P. gaudichaudianum para avaliação dos efeitos moluscicida. O eugenol e os óleos essenciais de O. majorana e O. americanum apresentaram efeito relaxante nas concentrações 250µL L-1, mas os óleos essenciais de H. ringens e P. gaudichaudianum não promoveram efeito relaxante ou moluscicida no tempo e concentrações estudadas. Portanto, apenas o eugenol e os óleos essenciais de O. majorana e O. americanum podem serem usados para fins de relaxamento em P. canaliculata.

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This study evaluated the potential relaxing and/or molluscicidal effects of eugenol and essential oils of Origanum majorana, Ocimum americanum, Hesperozygis ringens, and Piper gaudichaudianum in the gastropod Pomacea canaliculata. Compounds were tested at concentrations of 100, 250, 500, and 750µL L-1 to evaluate the relaxing effects. In the second experiment, animals were exposed to 10, 25, and 50µL L-1 of essential oils of H. ringens and P. gaudichaudianum for a period of 24h for the evaluation of molluscicidal effects. Eugenol and essential oils of O. majorana and O. americanum showed relaxing effects at 250µL L-1, but the essential oils of H. ringens and P. gaudichaudianum did not promote relaxing or molluscicidal effects within the times and concentrations studied. Therefore, only eugenol and the essential oils of O. majorana and O. americanum can be used for relaxation purposes in P. canaliculata.(AU)

Este estudo teve como objetivo avaliar os possíveis efeitos relaxantes e/ou moluscicidas do eugenol e óleos essenciais de Origanum majorana, Ocimum americanum, Hesperozygis ringens e Piper gaudichaudianum no gastrópode Pomacea canaliculata. Os compostos foram testados nas concentrações de 100, 250, 500 e 750µL L-1 para avaliar os efeitos relaxantes. Em um segundo experimento, os animais foram expostos a 10, 25 e 50µL L-1 por 24 horas aos óleos essenciais de H. ringens e P. gaudichaudianum para avaliação dos efeitos moluscicida. O eugenol e os óleos essenciais de O. majorana e O. americanum apresentaram efeito relaxante nas concentrações 250µL L-1, mas os óleos essenciais de H. ringens e P. gaudichaudianum não promoveram efeito relaxante ou moluscicida no tempo e concentrações estudadas. Portanto, apenas o eugenol e os óleos essenciais de O. majorana e O. americanum podem serem usados para fins de relaxamento em P. canaliculata.(AU)

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ABSTRACT: This study evaluated the potential relaxing and/or molluscicidal effects of eugenol and essential oils of Origanum majorana, Ocimum americanum, Hesperozygis ringens, and Piper gaudichaudianum in the gastropod Pomacea canaliculata. Compounds were tested at concentrations of 100, 250, 500, and 750µL L-1 to evaluate the relaxing effects. In the second experiment, animals were exposed to 10, 25, and 50µL L-1 of essential oils of H. ringens and P. gaudichaudianum for a period of 24h for the evaluation of molluscicidal effects. Eugenol and essential oils of O. majorana and O. americanum showed relaxing effects at ≥250µL L-1, but the essential oils of H. ringens and P. gaudichaudianum did not promote relaxing or molluscicidal effects within the times and concentrations studied. Therefore, only eugenol and the essential oils of O. majorana and O. americanum can be used for relaxation purposes in P. canaliculata.

RESUMO: Este estudo teve como objetivo avaliar os possíveis efeitos relaxantes e/ou moluscicidas do eugenol e óleos essenciais de Origanum majorana, Ocimum americanum, Hesperozygis ringens e Piper gaudichaudianum no gastrópode Pomacea canaliculata. Os compostos foram testados nas concentrações de 100, 250, 500 e 750µL L-1 para avaliar os efeitos relaxantes. Em um segundo experimento, os animais foram expostos a 10, 25 e 50µL L-1 por 24 horas aos óleos essenciais de H. ringens e P. gaudichaudianum para avaliação dos efeitos moluscicida. O eugenol e os óleos essenciais de O. majorana e O. americanum apresentaram efeito relaxante nas concentrações ≥250µL L-1, mas os óleos essenciais de H. ringens e P. gaudichaudianum não promoveram efeito relaxante ou moluscicida no tempo e concentrações estudadas. Portanto, apenas o eugenol e os óleos essenciais de O. majorana e O. americanum podem serem usados para fins de relaxamento em P. canaliculata.