Efficacy of essential oils from plants cultivated in the Amazonian Biome against gastrointestinal nematodes in sheep.

The excessive use of anthelmintics to control nematodes has resulted in anthelminthic resistance. Essential oils (EOs) are a rich source of bioactive molecules that can be assessed for their ability to control resistant parasite populations. The aims of this study were to screen EOs from 10 plant species in vitro for anthelmintic activity against Haemonchus contortus, evaluate the cytotoxicity of those EOs in a human immortalized keratinocyte cell line (HaCaT), and test the most promising EO candidate in vivo in Santa Inês sheep. The efficacy was investigated in vitro using an egg hatch test (EHT) and a larval development test (LDT). EO cytotoxicity was evaluated with the sulforhodamine-B assay. In the in vivo experiment, 28 Santa Inês sheep naturally infected were distributed into groups: G1-Mentha arvensis (EO5), 200 mg kg-1; G2-menthol, 160 mg kg-1; G3-negative control; and G4-positive control (monepantel). EO5, from M. arvensis (86.7% menthol), had the lowest LC50 and LC90 values in the EHT (0.10, 0.27 mg mL-1, respectively), good performance in the LDT (0.015, 0.072 mg mL-1, respectively), and the lowest cytotoxicity (190.9 µg mL-1) in HaCaT cells. In the in vivo test, a single dose of the EO5 (200 mg kg-1 BW) had an efficacy of approximately 50% on days 1, 14, and 21; however, values were not significantly from day 0. Conversely, pure menthol at a dose of 160 mg kg-1 BW showed no in vivo efficacy. This can be attributed to key factors related to bioavailability and pharmacology of terpenes in the host organism, as well as to the fact that menthol is mainly excreted as glucuronides in urine. Thus, further studies should be conducted with formulation systems that deliver bioactives directly to the abomasum, focusing on terpenes, whose excretion route is mainly via faeces.

Piper Essential Oils Inhibit Rhizopus oryzae Growth, Biofilm Formation, and Rhizopuspepsin Activity.

Piper is the largest genus of the Piperaceae family. The species of this genus have diverse biological activities and are used in pharmacopeia throughout the world. They are also used in folk medicine for treatment of many diseases in several countries including Brazil, China, India, Jamaica, and Mexico. In Brazil, Piper species are distributed throughout the national territory, making this genus a good candidate for biological activity screening. During our studies with Piper essential oils, we evaluated its activity against Rhizopus oryzae, the main agent of mucormycosis. The main compounds of seven Piper essential oils analyzed were Piper callosum-safrole (53.8%), P. aduncum-dillapiole (76.0%), P. hispidinervum-safrole (91.4%), P. marginatum-propiopiperone (13.2%), P. hispidum-γ-terpinene (30.9%), P. tuberculatum-(E)-caryophyllene (30.1%), and Piper sp.-linalool (14.6%). The minimum inhibitory concentration of Piper essential oils against R. oryzae ranged from 78.12 to >1250 µg/mL. The best result of total inhibition of biofilm formation was obtained with Piper sp. starting from 4.88 µg/mL. Considering the bioactive potential of EOs against planktonic cells and biofilm formation of R. oryzae could be of great interest for development of antimicrobials for therapeutic use in treatment of fungal infection.

Oxidation of monoterpenes in Protium heptaphyllum oleoresins.

Protium heptaphyllum (Burseraceae) oleoresins are rich in volatile monoterpenes, exhibiting a chemical composition that can be strongly altered with time. The present work aimed to discuss the temporal change of the volatile composition of these oleoresins, and search for related supporting evidence. Samples of P. heptaphyllum oleoresin were collected separately for fresh (n = 10) and aged (n = 8) oleoresins, with the essential oils obtained by hydrodistillation analyzed by GC-FID and GC-MS. Fresh oleoresins were characterized by a high content of terpinolene (28.2-69.7%), whereas aged ones contained large amounts of p-cymene (18.7-43.0%) and p-cymen-8-ol (8.2-31.8%). Multivariate analyses were performed based on the yield and major essential oil components to clearly demonstrate the existence of two subsets (fresh and aged oleoresins). In addition, an analysis of the partial genome sequencing of the species was carried out, producing the largest amount of data for the genus Protium. Subsequently, were searched for nucleotide sequences responsible for the enzymes involved in the biosynthesis of monoterpenes. Two hypotheses were formulated to understand the oxidation process during aging of the oleoresins: (i) a natural chemical oxidation of terpenes and (ii) an oxidation catalyzed by enzymes produced by microorganisms associated with the plant. The results suggested that terpinolene was most likely oxidized to p-cymene, which, in turn, was oxidized into p-cymen-8-ol during natural aging of the exudate due to abiotic factors.