Evaluation of the anti-mycobacterial activity and composition of Carlina acaulis L. root extracts.

Nontuberculous mycobacteria (NTM) have recently emerged as important bacterial pathogens of animals and humans. Of particular concern is the high level of antimicrobial resistance displayed by these organisms, which complicates treatment and potential successful outcomes. Here, we evaluated the potential of Carlina acaulis L. as a source of novel anti-mycobacterial agents. Our goal was to measure the activity of aqueous, ethanol, and chloroform C. acaulis root extracts against 99 NTM strains. GC-MS spectroscopy analyses were performed to deliver qualitative and quantitative data on the composition of C. acaulis extract. In our study, we have shown for the first time the activity of C. acaulis extracts against NTM. The highest activity was exhibited by the chloroform extract, which inhibited the growth of more than 90% of the strains at the dose of 100 µg/mL (MIC90 = 100 µg/mL). The results of the GC-MS analysis of the C. acaulis chloroform extract contributed to the identification of 37 compounds, with carlina oxide as the most representative compound (69.52%) followed by 3,4-dihydro-2H-phenanthren- -1-one (6.54%) and stigmast-5-en-3-ol (4.14%). Our results indicate that C. acaulis chloroform and ethanol extracts have potential for treatment of NTM infections and that this plant contains anti-mycobacterial compounds.

Parasiticidal effects of Tanacetum vulgare extract against Ichthyophthirius multifiliis.

Ichthyophthiriasis, which is caused by Ichthyophthirius multifiliis (Ich) infections, has a severe impact on productivity in freshwater aquaculture. These infections were previously treated effectively with malachite green, a compound that is now banned on fish farms due to its carcinogenicity. To find efficacious drugs to control Ich, flowers of tansy Tanacetum vulgare were evaluated for their antiprotozoal activity. Tanacetum vulgare extract significantly reduced the survival of Ich trophonts and theronts. In vitro, the extract killed all trophonts at 3200 mg l-1, terminated tomont reproduction at 50 mg l-1, and caused mortality of all theronts at 100 mg l-1. T. vulgare extract may be a new and efficacious drug for the control of Ich.

Anti-babesial potential and chemical composition of essential oil from yarrow Achillea millefolium.

Essential oils from plants used in traditional medicine are known as a rich source of chemically diverse compounds with specific biological activities. Achillea millefolium essential oil (AEO) was screened for in vitro activity against Babesia canis. The AEO was obtained by hydrodistillation and analysed by gas chromatography coupled to mass spectrometry (GC-MS). GC-MS revealed the presence of 47 compounds in the essential oil. Those present in the highest concentrations were chamazulene (34.45%), ß-caryophyllene (8.93%), (E)-germacrene D (7.55%), patchoulene (7.27%), ß-guaiene (4.62%), α-humulene (4.59%), santolina epoxide (4.41%), ethyl iso-allocholate (2.97%), aromadendrene (2.62%), and neoclovenoxid-alkohol (2.46%). AEO was found to be active in vitro against B. canis, with 50% inhibitory concentration (IC50) values of 0.06 mg/mL, as compared to imidocarb, with IC50 = 0.007 mg/mL. The study confirms that essential oil from A. millefolium has anti-babesial properties in vitro.

Inhibitory activities of essential oils against Babesia canis.

The in vitro anti-Babesia canis activities of nine essential oils were investigated. Among the tested essential oils Achillea millefolium, Eugenia caryophyllus and Citrus grandis were the most active (IC50 values of 51.0, 60.3 and 61.3 µg/mL, respectively). The oils from Abies sibirica, Rosmarinus officinalis, Eucalyptus globulus, Cinnamonum zeylanicum, Mentha piperita and Pinus sylvestris were less active (IC50 values of 134.3, 237.3, 239.3, 367.9, 837.5 and 907.3 µg/mL, respectively). The results support the concept that some essential oil constituents may be useful in the clinical management of babesiosis.

In vitro antioxidant and antibabesial activities of the extracts of Achillea millefolium.

Despite many phytochemical and pharmacological investigations, to date, there are no reports concerning the antibabesial activity of extracts of A. millefolium against B. canis. This study was aimed at investigating the biological activities of A. millefolium against the Babesia canis parasite and to identify its chemical ingredients. The water (WE), ethanol (EE) and hexane/acetone (H/AE) extracts of plant aerial parts were screened for total phenolic content (TPC), total flavonoid compound (TFC), DPPH free radical-scavenging activity and its antibabesial activity assay. In this study, imidocarb diproprionate was used as a positive control. The H/AE and EE extracts were analysed using gas chromatography-mass spectroscopy (GC-MS). In the EE extract, the main compounds were 17.64% methyl octadec-9-ynoate, 16.68% stigmast-5-en-3-ol(3α,24S) and 15.17% hexadecanoic acid. In the H/AE extract, the main compounds were 34.55% 11-decyldocosane, 14.31% N-tetratetracontane, 8.22% ß-caryophyllene, and 7.69% N-nonacosane. Extract of EE contained the highest content of phenolics followed by H/AE and WE. The concentration of flavonoids in EE, H/AE and WE extracts showed that TFC was higher in the EE samples followed by H/AE and WE. The antioxidant activities were highest for AA, followed by EE, WE and H/AE. The antibabesial assay showed that the WE, EE and H/AE extracts of A. millefolium were antagonistic to B. canis. At a 2 mg/mL concentration, it showed 58.7% (± 4.7%), 62.3% (± 5.5%) and 49.3% (± 5.1%) inhibitory rate in an antibabesial assay, respectively. Considering these results, the present findings suggest that A. millefolium extracts may be a potential therapeutic agent and that additional studies including in vivo experiments are essential.

Effect of dietary supplementation with Echinacea purpurea on vaccine efficacy against infection with Flavobacterium columnare in zebrafish (Danio rerio).

The effect of dietary Echinacea purpurea (EP) on the response of zebrafish (Danio rerio) to a Flavobacterium columnare vaccine was investigated. Two hundred D. rerio with an average weight of 290 ± 40 g were selected and fed different levels of E. purpurea (5 g kg(-1) diet--group 1, 10 g kg(-1) diet--group 2, 20 g kg(-1) diet--group 3, 30 g kg(-1) diet--group 4, and 0 g kg(-1) diet--group 5). Experimental feeding was begun 3 weeks prior to bath immunization and continued until the end of the experiment. Twenty-eight days after immunization the fish were challenged by bath immersion with F. columnare at a concentration of 1 x 10(6) CFU/ml. The relative percent survival of the experimental groups (1, 2, 3, 4 and 5) was 5.0, 6.0, 30.0, 36.0 and 5.0, respectively. In conclusion, diet supplementation with E. purpurea may effectively enhance the response of zebrafish to a F. columnare vaccine.

Humoral antibody response of Atlantic salmon, Salmo salar L., vaccinated against Moritella viscosa.

Immunization of Atlantic salmon, Salmo salar L., with experimental vaccine for protection against M. viscosa was tested. The antigen preparation used for the comparative studies was sonicate formalin killed M. viscose unwashed bacteria. Vaccination was carried out by intraperitoneal injection of bacterin in Biojec mineral oil as an adjuvant. When accumulated mortality in the control group with PBS of fish challenged by i.p. injection of M. viscose reached 100% with a challenge dose of 2.3 x 10(6) CFU, mortality in the experimental group was 71.23%. A positive correlation between the survival rates and corresponding median antibody levels of each group was found at the challenge time.