Cold-Pressed Sacha Inchi Oil: High in Omega-3 and Prevents Fat Accumulation in the Liver.

The ability of oil supplementation to inhibit various metabolic syndromes has been recognized. However, there are currently no studies determining the effects of oil supplements on healthy conditions. Plukenetia volubilis L., also known as Sacha inchi, is a seed rich in essential unsaturated fatty acids that improves metabolic syndrome diseases, such as obesity and nonalcoholic fatty liver. However, the health benefits and effects of Sacha inchi oil (SIO) supplementation remain unclear. This study aims to evaluate the chemical effects and properties of Sacha inchi oil. The results of the chemical compound analysis showed that Sacha inchi is an abundant source of ω-3 fatty acids, with a content of 44.73%, and exhibits scavenging activity of 240.53 ± 11.74 and 272.41 ± 6.95 µg Trolox/g, determined via DPPH and ABTS assays, respectively, while both olive and lard oils exhibited lower scavenging activities compared with Sacha inchi. Regarding liver histology, rats given Sacha inchi supplements showed lower TG accumulation and fat droplet distribution in the liver than those given lard supplements, with fat areas of approximately 14.19 ± 6.49% and 8.15 ± 2.40%, respectively. In conclusion, our findings suggest that Sacha inchi oil is a plant source of ω-3 fatty acids and antioxidants and does not induce fatty liver and pathology in the kidney, pancreas, and spleen. Therefore, it has the potential to be used as a dietary supplement to improve metabolic syndrome diseases.

The combined recombinant cathepsin L1H and cathepsin B3 vaccine against Fasciola gigantica infection.

Protections against Fasciola gigantica infection in mice immunized with the individual and combined cathepsin L1H and cathepsin B3 vaccines were assessed. The vaccines comprised recombinant (r) pro-proteins of cathepsin L1H and B3 (rproFgCatL1H and rproFgCatB3) and combined proteins which were expressed in Pichia pastoris. The experimental trials were performed in ICR mice (n = 10 per group) by subcutaneous injection with 50 µg of the recombinant proteins combined with Alum or Freund's adjuvants. At two weeks after the third immunization, mice were infected with 15 F. gigantica metacercariae per mouse by oral route. The percents of protection of rproFgCatL1H, rproFgCatB3 and combined vaccines against F. gigantica were approximately 58.8 to 75.0% when compared with adjuvant-infected control. These protective effects were similar among groups receiving vaccines with Alum or Freund's adjuvants. By determining the levels of IgG1 and IgG2a in the immune sera, which are indicative of Th1 and Th2 immune responses, it was found that both Th1 and Th2 humoral immune responses were significantly increased in vaccinated groups compared with the control groups, with higher levels of IgG1 (Th2) than IgG2a (Th1). Mice in vaccinated groups showed reduction in liver pathological lesions when compared with control groups. This study indicates that the combined rproFgCatB3 and rproFgCatL1H vaccine had a high protective potential than a single a vaccine, with Alum and Freund's adjuvants showing similar level of protection. These results can serve as guidelines for the testing of this F. gigantica vaccine in larger economic animals.

A novel Thioredoxin-related protein 14 from Fasciola gigantica has an immunodiagnostic potential for fasciolosis.

In the definitive host, a trematode parasite can survive and evade the damage by reactive oxygen species that are generated from its metabolism and the host immune cells. Several anti-oxidant proteins are found in Fasciola spp. which play essential roles in cellular redox balance. One of them is thioredoxin-related protein 14 (TRP14) that has a highly conserved WCPDC motif and serves as a disulfide reductase-like thioredoxin (Trx). In the present study, a cDNA encoding TRP14 from F. gigantica (FgTRP14) was selected and cloned by immunoscreening with a rabbit infected serum. Phylogenetic analysis was performed by MEGA X program showed that FgTRP14 was most highly related to the Fasciola hepatica. Immunoblotting analysis of the polyclonal antibody rabbit serum against recombinant FgTRP14 (rFgTRP14) revealed that the molecular weight of natural FgTRP14 was at 14 kDa from metacercariae, NEJ, 4-week old juvenile and adult stage. The native FgTRP14 was expressed in caecal epithelial cells and preferentially localized on the cells' surface lamellae of adult stage. By sandwich ELISA assay, the circulating FgTRP14 could be recognized in sera of experimentally F. gigantica metacercariae infection in mice. The native FgTRP14 in the excretory-secretory (ES) and whole body (WB) of adult F. gigantica were detected at the concentrations 6.3 ng/ml, and 45 ng/ml, respectively. Therefore, it could be considered for immunodiagnostic candidate for fasciolosis.

Monoclonal antibody against Fasciola gigantica glutathione peroxidase and their immunodiagnosis potential for fasciolosis.

Glutathione peroxidases (GPx), major antioxidant enzymes, secreted by Fasciola spp., are important for the parasite evasion and protection against the host's immune responses. In the present study, a monoclonal antibody (MoAb) against recombinant F. gigantica glutathione peroxidase (rFgGPx) was produced by hybridoma technique using spleen cells from BALB/c mice immunized with rFgGPx. This MoAb (named 7B8) is IgG1 with κ light chains, and it reacted specifically with rFgGPx at a molecular weight 19 kDa as shown by immunoblotting, and reacted with the native FgGPx in the extracts of whole body (WB), metacercariae, newly excysted juveniles (NEJs), 4 week-old juveniles and adult F. gigantica as shown by indirect ELISA. It did not cross react with antigens in WB fractions from other adult trematodes, including Fischoederius cobboldi, Paramphistomum cervi, Setaria labiato-papillosa, Eurytrema pancreaticum, Gastrothylax crumenifer and Gigantocotyle explanatum. By immunolocalization, MoAb against rFgGPx reacted with the native protein in the tegument, vitelline cells, and eggs of adult F. gigantica. In addition, the sera from mice experimentally infected with F. gigantica were tested positive by this indirect sandwich ELISA. This result indicated that FgGPx is an abundantly expressed parasite protein that is secreted into the tegumental antigens (TA), therefore, FgGPx and its MoAb may be used for immunodiagnosis of both early and late fasciolosis gigantica in animals and humans.

Expression and characterization of glutathione peroxidase of the liver fluke, Fasciola gigantica.

Glutathione peroxidase (GPx) is a key member of the family of antioxidant enzymes in trematode parasites including Fasciola spp. Because of its abundance and central role as an anti-oxidant that helps to protect parasites from damage by free radicals released from the host immune cells, it has both diagnostic as well as vaccine potential against fasciolosis. In this study, we have cloned, characterized, and detected the expression of the GPx protein in Fasciola gigantica (Fg). FgGPx (582 bp) was cloned by polymerase chain reaction (PCR) from complementary DNA (cDNA) from an adult fluke. Its putative peptide has no signal sequence and is composed of 168 amino acids, with a molecular weight of 19.1 kDa, and conserved sequences at NVACKUG, FPCNQFGGQ, and WNF. Phylogenetic analysis showed that GPx is present from protozoa to mammals and FgGPx was closely related to Fasciola hepatica GPx. A recombinant FgGPx (rFgGPx) was expressed in Escherichia coli BL21 (DE3) and used for immunizing mice to obtain polyclonal antibodies (anti-rFgGPx) for immunoblotting and immunolocalization. In immunoblotting analysis, the FgGPx was expressed in all stages of F. gigantica (eggs, metacercariae, newly excysted juveniles (NEJ), 4-week-old juveniles, and adults). This mouse anti-rFgGPx reacted with the native FgGPx at a molecular weight of 19.1 kDa in adult whole body (WB) and tegumental antigens (TA) as detected by immunoblotting. The FgGPx protein was expressed at a high level in the tegument, vitelline glands, and eggs of the parasite. Anti-rFgGPx exhibited no cross-reactivity with the other parasite antigens, including Eurytrema pancreaticum, Cotylophoron cotylophorum, Fischoederius cobboldi, Gastrothylax crumenifer, Paramphistomum cervi, and Setaria labiato papillosa. The possibility of using rFgGPx for immunodiagnosis and/or as a vaccine for fasciolosis in animals of economic importance will be explored in the future.