The Use of Essential Oil and Hydrosol Extracted from Cuminum cyminum Seeds for the Control of Meloidogyne incognita and Meloidogyne javanica

The essential oil (EO) and hydrosol (HL) isolated from Cuminum cyminum (cumin) seeds were evaluated against the root-knot nematodes Meloidogyne incognita and M. javanica. The efficacy of extracts on the motility, hatching, and survival in soil of second-stage juveniles (J2s), and the activity on egg differentiation were tested. All J2s were paralyzed after immersion in the EO at 62.5 μL/L concentration for 96 h. Encouraging results were recorded using HL equal to or higher than 10% concentration for both Meloidogyne species tested. More than 70% paralyzed J2s were recorded after immersion for 48 h, while the percentage was increased to higher than 90% after 96 h of immersion. A clear effect on egg differentiation was observed after immersion in EO or HL. A significant decrease in egg differentiation was revealed at even low concentrations of EO while an evident decrease in egg differentiation was recorded after immersion of eggs in 50% HL dilution. Decreased hatching of M. incognita and M. javanica J2s was observed with the increase in concentration. The lowest numbers of hatched J2s were recorded when EO was used at 1000 and 2000 μL/L concentrations. A constant reduction in root-knot nematode J2 hatching was observed upon increasing the concentration of HL from 5% to 50%. The EO of C. cyminum is characterized by the presence of γ-terpinene-7-al (34.95%), cumin aldehydes (26.48), and α-terpinene-7-al (12.77%). The above constituents were observed in HL following the same order as that observed in EO. The components γ-terpinene (11.09%) and ο-cymene (6.56%) were also recorded in EO while they were absent in HL

Additional file 1: Figure S1. of Nogo receptor complex expression dynamics in the inflammatory foci of central nervous system experimental autoimmune demyelination

Bielschowsky staining, immunofluorescence, and real-time PCR quality controls. (A1–4) Typical images of scores 0–3 in Bielschowsky silver staining. (B, C) Preabsorption assay for LINGO-1 and TROY in serial sections of chronic and acute phases, respectively (the phase where signal is most abundant). (B1) LINGO-1 antibody specifications: rabbit polyclonal, Abcam ab23631, LOT N/A, dilution 1:300. (B2) LINGO-1 peptide specifications: rabbit, Abcam ab25890, LOT #GR41007-1, incubation with ab in 10× molecular ratio. (C1) TROY antibody specifications: goat polyclonal, Santa Cruz sc-13711 (E-19), LOT #H0707, epitope mapping near the C-terminus of TROY of mouse origin, dilution 1:100. (C2) TROY peptide specifications: goat, Santa Cruz sc-13711 P, LOT #B0402, incubation with ab in 10× molecular ratio. (D, E) Antibody specificity test for p75 and NgR with the use of another antibody (different company) recognizing a different epitope in serial sections of chronic and acute phases, respectively (the phase where signal is most abundant). (D1) p75 antibody #1 specifications: mouse monoclonal, Abcam ab8877, LOT GR136825-1, ME20.4, dilution 1:400. (D2) p75 antibody #2 specifications: mouse monoclonal, Santa Cruz p75 (B-1) sc-271708, LOT #J0611, epitope mapping between amino acids 393–427 at the C-terminus of NGFR p75 of human origin, 1:100. (E1) NgR antibody #1 specifications: rabbit polyclonal, Santa Cruz sc-25659 (H-120), LOT E1209, epitope corresponding to amino acids 31–150 mapping near the N-terminus of Nogo-R of human origin, dilution 1:100. (E2) NgR antibody #2 specifications: rabbit polyclonal, Abcam ab26291, LOT N/A, epitope from within residues 150–250 of rat Nogo receptor, dilution 1:100. (F) β-actin real-time PCR quality control showing the specific amplification products on agarose gel and the melting curves of their respective genes; curve identifier: light green TROY, orange p75, dark green LINGO-1. (G) mRNA levels of coreceptors LINGO-1, p75, and TROY in the spinal cord of EAE animals by real-time PCR analysis using GAPDH as a second, quality control, house-keeping gene. The levels of mRNA expression for the coreceptors (G1) LINGO-1, (G2) p75, and (G3) TROY, followed the same pattern with those that underwent β-actin analysis. Error bars indicate the standard statistical error of the mean (SEM), ***p < 0.001, **p < 0.01. Black scale bar = 20 μm