GABAa receptor subunits expression in silver catfish (Rhamdia quelen) brain and its modulation by Nectandra grandiflora Nees essential oil and isolated compounds

Studies using silver catfish (Rhamdia quelen) as experimental models are often applied to screen essential oils (EO) with GABAergic-mediated effects. However, the expression of GABAa receptors in the silver catfish brain remains unknown. Thus, we assessed whether silver catfish express GABAa receptor subunits associated with sedation/anesthetic process and/or neurological diseases. Additionally, we evaluated the brain expression of GABAa receptor subunits in fish sedated with Nectandra grandiflora EO and its isolated compounds, the fish anesthetic (+)-dehydrofukinone (DHF), and dehydrofukinone epoxide (DFX), eremophil-11-en-10-ol (ERM) and selin-11-en-4-α-ol (SEL), which have GABAa-mediated anxiolytic-like effects in mice. The expression of the subunits gabra1, gabra2, gabra3, gabrb1, gabrd and gabrg2 in the silver catfish brain were assessed after a 24h-sedation bath by real time PCR. Since qPCR data rarely describes mechanisms of action, which are usually found through interactions with receptors, we also performed an antagonist-driven experiment using flumazenil (FMZ). Real-time PCR detected the mRNA expression of all targeted genes in R. quelen brain. The expression of gabra1 was decreased in fish sedated with ERM; EO increased gabra2, gabra3, gabrb1 and gabrg2 expression; SEL increased gabrb1, gabrd and gabrg2 expression. EO and compounds DFX, SEL and ERM induced sustained sedation in fish and FMZ-bath prompted the recovery from ERM- and DFX-induced sedation. Our results suggest that the EO, SEL, ERM and DFX sedative effects involve interaction with the GABAergic system. Our findings support the use of the silver catfish as robust and reliable experimental model to evaluate the efficacy of drugs with putative GABAergic-mediated effects.This work was supported in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brazil (CAPES) - Finance Code 001 and by the National Council of Scientific and Technological Development (CNPq, Brazil) scholarship. BMH and BB are recipients of productivity fellowships of CNPq (grant numbers 454447/2014-0 and 301156/2012-3, respectively)

The Use of Cinnamon Essential Oils in Aquaculture: Antibacterial, Anesthetic, Growth-Promoting, and Antioxidant Effects

Cinnamon essential oils (EOs) are widely known for their pharmaceutical properties; however, studies investigating the use of these EOs in aquaculture are scarce. The aims of this study were to evaluate the anesthetic effect of bathing silver catfish (Rhamdia quelen) with Cinnamomum cassia EO (CCEO) and its nanoemulsion (NCCEO); the growth-promoting and antioxidant effects of dietary supplementation with CCEO in silver catfish; and the in vitro antibacterial effect of CCEO, NCCEO, and Cinnamomum zeylanicum EO (CZEO) against bacteria isolated from diseased silver catfish. The two cinnamon EOs showed promising antibacterial activity, which was potentiated by the nanoemulsion. CCEO showed satisfactory anesthetic activity in silver catfish, and its nanoemulsion intensified the sedative activity. Supplementation of 1.0 mL CCEO per kg of diet for 60 days increased weight, length, and weight gain when compared to the control group, evidencing the growth-promoting activity of this EO. Dietary supplementation of CCEO for 30 and 60 days also showed an antioxidant effect, as it decreased levels of thiobarbituric acid reactive species and increased the superoxide dismutase activity in the liver of silver catfish. Therefore, cinnamon EOs have a promising use in aquaculture

The Use of Cinnamon Essential Oils in Aquaculture: Antibacterial, Anesthetic, Growth-Promoting, and Antioxidant Effects

Cinnamon essential oils (EOs) are widely known for their pharmaceutical properties; however, studies investigating the use of these EOs in aquaculture are scarce. The aims of this study were to evaluate the anesthetic effect of bathing silver catfish (Rhamdia quelen) with Cinnamomum cassia EO (CCEO) and its nanoemulsion (NCCEO); the growth-promoting and antioxidant effects of dietary supplementation with CCEO in silver catfish; and the in vitro antibacterial effect of CCEO, NCCEO, and Cinnamomum zeylanicum EO (CZEO) against bacteria isolated from diseased silver catfish. The two cinnamon EOs showed promising antibacterial activity, which was potentiated by the nanoemulsion. CCEO showed satisfactory anesthetic activity in silver catfish, and its nanoemulsion intensified the sedative activity. Supplementation of 1.0 mL CCEO per kg of diet for 60 days increased weight, length, and weight gain when compared to the control group, evidencing the growth-promoting activity of this EO. Dietary supplementation of CCEO for 30 and 60 days also showed an antioxidant effect, as it decreased levels of thiobarbituric acid reactive species and increased the superoxide dismutase activity in the liver of silver catfish. Therefore, cinnamon EOs have a promising use in aquaculture

Involvement of HPI-axis in anesthesia with Lippia alba essential oil citral and linalool chemotypes: gene expression in the secondary responses in silver catfish

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.Authors are grateful to the Conselho Nacional de Desenvolvimento Tecnológico (CNPq), Comissão de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and INCT-ADAPTA 2 (CNPq)

Expression of Ion Transporters and Na+/K+-ATPase and H+-ATPase Activities in the Gills and Kidney of Silver Catfish (Rhamdia quelen) Exposed to Different pHs

Exposure to acidic and alkaline pHs results in an ionic imbalance. Cellular responses involved in osmoregulation in silver catfish exposed to different pHs (5.5, 7.5, and 9.0) for 24 h were evaluated. The gills and kidney were collected to measure Na+/K+-ATPase (NKA) and H+-ATPase (V-ATPase) activities and to evaluate the expression of ion transporter-related genes: NKA (atp1a1), H+-ATPases (atp6v0a1b, atp6v0a2a, atp6v0a2b), Na+/H+ antiporter (slc9a3), K+/Cl− symporters (slc12a4, slc12a6, slc12a7a, slc12a7b), Na+/K+/2Cl− symporter (slc12a2), and ammonium transporter Rh type b (rhbg). The gills presented greater responses to pH changes than the kidney. The pH alterations changed the atp1a1 gene expression and NKA activity, whereas the H+-ATPase activity increased in the gills in alkaline water, probably to maintain ionic balance. The slc9a3 and slc12a2 genes play more prominent roles in the ion uptake at acidic pH than H+-ATPase. The slc12a7a was the only isoform of this transporter affected by pH. The rhbg is apparently related to ammonia excretion through the gills and kidney (minor scale). Exposure to alkaline pH seems to be battled by impairment of NKA and H+-ATPase activities in the gills, whereas the expression of some ion transporters in silver catfish changes during both acidic and alkaline pHs

Plant essential oils against bacteria isolated from fish: an in vitro screening and in vivo efficacy of Lippia origanoides

ABSTRACT: The use of natural products, such as essential oils (EOs), is a potential novel approach to treat fish bacterial infections with a lower risk of developing resistance. There has been a number of studies reporting the activity of EOs as those obtained from the species Achyrocline satureioides, Aniba parviflora, Aniba rosaeodora, Anthemis nobilis, Conobea scoparioides, Cupressus sempervirens, Illicium verum, Lippia origanoides, and Melaleuca alternifolia against bacteria. However, there are few studies investigating the effect of these EOs against fish bacteria. Therefore, the aim of this study was to evaluate the in vitro antibacterial activity of EOs against the following fish bacteria, Aeromonas hydrophila, Citrobacter freundii, and Raoultella ornithinolytica. Additionally, the in vivo antibacterial activity of the EO L. origanoides was evaluated against experimentally induced A. hydrophila infection of silver catfish (Rhamdia quelen). The EO of L. origanoides was chosen as it showed the highest in vitro antibacterial activity, with minimum inhibitory concentrations ranging from 0.2 to 0.8 mg mL-1. This EO also presented a therapeutic success of 58.33%, on a 30 day A. hydrophila infection. Therefore, we suggested that the EO of L. origanoides may be a viable alternative as a treatment for A. hydrophila infection