Enhancement of Bioactivity of Natural Extracts by Non-Thermal High Hydrostatic Pressure Extraction

Natural extracts, like those obtained from medicinal herbs, dietary plants and fruits are being recognized as important sources of bioactive compounds with several functionalities including antioxidant, anticancer, and antimicrobial activities. Plant extracts rich in phenolic antioxidants are currently being successfully used for several pharmaceutical applications and in the development of new foods (i.e., functional foods), in order to enhance the bioactivity of the products and to replace synthetic antioxidants. The extraction method applied in the recovery of the bioactive compounds from natural materials is a key factor to enhance the bioactivity of the extracts. However, most of the extraction techniques have to employ heat, which can easily lead to heat-sensitive compounds losing their biological activity, due to changes caused by temperature. Presently, high hydrostatic pressure (HHP) is being increasingly explored as a cold extraction method of bioactive compounds from natural sources. This non-thermal high hydrostatic pressure extraction (HHPE) technique allows one to reduce the extraction time and increase the extraction of natural beneficial ingredients, in terms of nutritional value and biological activities and thus enhance the bioactivity of the extracts. This review provides an updated and comprehensive overview on the extraction efficiency of HHPE for the production of natural extracts with enhanced bioactivity, based on the extraction yield, total content and individual composition of bioactive compounds, extraction selectivity, and biological activities of the different plant extracts, so far studied by extraction with this technique.     

The effect of adenosine on pro-inflammatory cytokine production by porcine T cells

Adenosine is a well described anti-inflammatory modulator of immune responses. The aim of the present study was to describe the role of common adenosine agonist 5'-N-ethylcarboxamidoadenosine (NECA) in cytokine production by main porcine T cell subpopulations. TNF-α, IFN-γ, IL-2 and IL-10 were detected by multicolor flow cytometry together with cell surface markers CD3, CD4 and CD8. It was found that NECA inhibits (in a dose-dependent manner) production of pro-inflammatory TNF-α and Th1-associated cytokines IFN-γ, IL-2 in all concanavalin A-stimulated T cell subpopulations. Moreover, production of IL-10 was potentiated in all T cell subpopulations tested. These corresponded well with the fact that all T cell subsets expressed mRNA for adenosine receptor (AR) subtypes to comparable extents. Contrary to concanavalin A-stimulated cells, NECA had a moderate effect on PMA-stimulated T cells, suggesting that AR in pigs acts via signaling pathways not associated with protein-kinase C. Non-selective antagonist CGS15943 as well as allosteric modulator SCH202676 failed to reverse the effect of NECA in pigs. In conclusion, NECA has an anti-inflammatory effect on porcine T cell subpopulations.     

Import and efflux of flubendazole in Haemonchus contortus strains susceptible and resistant to anthelmintics

Drug entry into the body of a helminth is a key factor in the efficacy of anthelmintics. The present project was designed to study the ex vivo uptake and efflux of the benzimidazole anthelmintic flubendazole (FLU) in four strains of H. contortus: the ISE strain (fully susceptible to anthelmintics), the ISE-S strain (resistant to ivermectin), the BR strain (resistant to benzimidazoles) and the WR strain (multi-resistant). The transport of FLU between dead and living nematodes was also compared as well as the effect of verapamil, an inhibitor of the main efflux ABCB1 transporter (P-glycoprotein), on FLU accumulation in nematodes. The obtained results showed that FLU is able to effectively enter H. contortus adults due to high FLU lipophilicity. Passive diffusion is probably the only mechanism in both FLU import and efflux from nematodes. No differences in FLU transport were found among four H. contortus strains with different sensitivity to anthelmintics. No active FLU efflux from H. contortus and no effect of verapamil were observed, indicating that H. contortus cannot protect itself against FLU by the active removal of this anthelmintic from its body.     

Sample size calculations for Bayesian prediction of bovine viral-diarrhoea-virus infection in beef herds

We used a Bayesian classification approach to predict the bovine viral-diarrhoea-virus infection status of a herd when the prevalence of persistently infected animals in such herds is very small (e.g. <1%). An example of the approach is presented using data on beef herds in Wyoming, USA. The approach uses past covariate information (serum-neutralization titres collected on animals in 16 herds) within a predictive model for classification of a future observable herd. Simulations to estimate misclassification probabilities for different misclassification costs and prevalences of infected herds can be used as a guide to the sample size needed for classification of a future herd.     

The efficacy of three mycotoxin adsorbents to alleviate aflatoxin B1-induced toxicity in Oreochromis niloticus

Aflatoxicosis, toxicity of aflatoxin, is of great concern in aquaculture. This study was conducted to assess the efficacies of three adsorbents, a hydrated sodium calcium aluminosilicates (HSCAS), Saccharomyces cerevisiae (S.C.) and an esterified glucomannan (EGM), against feed contaminated with contained 200 μg/kg (ppb) aflatoxin B₁ (AFB₁). A total of 240 Nile tilapia fingerlings, Oreochromis niloticus (15 ± 2 g), were randomly divided into eight experimental groups (30 fish per group) with three replicates. Group T₁ represented the negative control fed on a basal diet, and T₂ was the positive control group fed on a basal diet supplemented with 200 ppb AFB₁. Groups T₃, T₄ and T₅ were fed the AFB₁-contaminated diet (200 ppb) supplemented with 0.5 % HSCAS, 0.25 % S.C or 0.25 % EGM, respectively. Groups T₆, T₇ and T₈ were fed a basal diet supplemented with 0.5 % HSCAS, 0.25 % S.C or 0.25 % EGM, respectively. The reduction in AFB₁-bioavailability was judged by toxin residues in fish musculature throughout the study beginning at the second week of exposure. AFB₁ reduced the survivability, total weight gain, average daily gain and specific growth rate, evident as early as the second week of exposure. The total erythrocyte count, hemoglobin content and total leukocyte count were significantly decreased after AFB₁ exposure for 6, 8 and 10 weeks, respectively. Prolonged administration of AFB₁ led to significant increases in serum alanine transaminase, aspartate transaminase and creatinine activity, and produced significant decreases in plasma proteins, including serum globulin. The specific immune response was assessed by an agglutinating antibody titer after immunization of the fish with an Aeromonas hydrophila vaccine. The antibody titer and relative level of protection of fish challenged with Aeromonas hydrophila were reduced throughout the period of examination in AFB₁-exposed fish. Supplementation with HSCAS, S.C. or EGM significantly improved growth performance, blood parameters and immune status; in addition, these groups showed decreased AFB₁ residues in fish musculature when compared with AFB₁-treated fish. HSCAS effectively reduced AFB₁ toxicity, whereas S.C. and EGM were less efficacious.