In Vitro Antimicrobial and Antibiofilm Activity of S-(-)-Limonene and R-(+)-Limonene against Fish Bacteria

Brazilian fish farming goes together with the emergence of numerous bacterial diseases, with Aeromonas hydrophila being the main bacterial pathogen. As a consequence, antimicrobials are excessively used. Considering that antimicrobials are relatively stable and nonbiodegradable, medicinal plants and their phytochemicals have been used as alternative treatments of bacteriosis in fish farming. Limonene is a monoterpene available in two enantiomers: R-(+)-limonene and S-(-)-limonene. This study analyzed the antibacterial activity of the phytochemicals S-(-)-limonene and R-(+)-limonene against some bacteria isolated from silver catfish (Rhamdia quelen). Furthermore, by means of spectrophotometry and atomic force microscopy, we also investigated the combination therapy of phytochemicals with antimicrobials and their activity in terms of inhibiting biofilm formation. Six clinical isolates and a standard strain were selected for antimicrobial activity testing. Biofilm formation was tested in 96-well plates and nylon cubes. The most sensitive of the strains tested was the A. hydrophila strain (MF 372510). S-(-)-limonene and R-(+)-limonene had high minimum inhibitory concentrations; however, they strongly inhibited A. hydrophila biofilm formation. R-(+)-limonene and S-(-)-limonene had an additive effect when combined with florfenicol and an antagonistic effect with oxytetracycline. In general, the phytochemicals tested showed strong antibiofilm activity against A. hydrophila, and when in combination therapy with florfenicol, they showed an additive effect against the treatment of A. hydrophila

Temporal profile of antimicrobial resistance exhibited by strains of Staphylococcus spp. isolated from cases of bovine mastitis for 20 years (1992-2011)

Records of in vitro susceptibility tests performed between 1992 and 2011 were retrospectively reviewed in order to evaluate the dynamic profiles of possible changes in antimicrobial resistance of Staphylococcus spp. isolated from milk samples of cows with mastitis during two decades. The results of 2,430 isolates tested by disk diffusion technique for susceptibility to oxacillin, penicillin, ampicillin, cephalexin, norfloxacin, tetracycline, sulfazotrim, gentamicin, and neomycin were analysed. Comparisons were performed between the percentages of resistance to antimicrobials and their classes and also between the decades studied. Additionally, the possible tendency or changes in the behaviour of these pathogens against the major drugs used in the last two decades were evaluated using regression analysis. The highest rates of resistance (P<0.0001) were observed for the beta-lactams (34.3%), with exception of cephalexin (6.9%), and for the tetracyclines (28%). Similar resistance rates (7.6% to 15.7%) were observed among the other drugs. Regression analysis showed a reduction in resistance to penicillin and ampicillin throughout the period, whilst for oxacillin and neomycin a decrease in the resistance was observed during the first decade, followed by an increase. A trend towards decreased resistance was found for sulfazotrim, whereas for the other antimicrobials no decrease was observed. The results indicated no trend towards increased resistance for most antimicrobials tested. Nevertheless, it is necessary to monitor the resistance patterns of these pathogens in order to save these drugs as a therapeutic reserv

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

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

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

ABSTRACT 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

Maclura tinctoria Extracts: In Vitro Antibacterial Activity against Aeromonas hydrophila and Sedative Effect in Rhamdia quelen

Maclura tinctoria is a tree species native from Brazil and rich in phenolic compounds. Since plant antibacterial activity is highly associated with phenolic compound concentration, we aim to evaluate the in vitro antimicrobial activity of different extracts against fish pathogenic bacteria. In addition, some phenolic compounds have central depressant effects and can be useful in aquaculture due to possible sedative and/or anesthetic effects. Four M. tinctoria extracts were extracted separately with ethanol; leaves (LE), bark (BE), heartwood (HE), and the sapwood (SE). In vitro antimicrobial activity was tested against Aeromonas strains at concentrations of 6400 to 3.125 μg/mL. The sedative effect was evaluated for 24 h with 30 and 100 mg/L concentrations. Chemical composition was analyzed by HPLC-DAD-MS. The HE extract had the best MIC (400 µg/mL) and MBC (800 µg/mL) compared to the LE, BE, and SE extracts. LE extract induced deep sedation and the BE, SE, and HE extracts induced light sedation. Additionally, BE, SE, and HE induced a normal behavior without side effects. Polyphenolic compounds with antimicrobial activity and sedative effects were identified mainly in HE. Thus, HE extract is safe and can be used as a sedative for silver catfish