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 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.     

New data on aetiology of nodular gill disease in rainbow trout, Oncorhynchus mykiss

We studied amoebae associated with nodular gill disease (NGD) outbreaks in rainbow trout Oncorhynchus mykiss (Walbaum) in fish farms in South-Western Germany. Gills of 12 diseased rainbow trout were examined in fresh, by isolation attempts, histologically and using in situ hybridisation (ISH). A total of nine amoeba strains of the genera Acanthamoeba (1), Hartmannella (2), Naegleria (1), Protacanthamoeba (1) and Vannella (4) were isolated and determined using light microscopical, ultrastructural and molecular methods. Specific molecular probes designed from the SSU rDNA sequences of individual amoeba strains were used for non-radioactive ISH in histological sections. Association of Naegleria sp. with NGD and a direct ISH proof of Naegleria trophozoites attached to hyperplastic gill epithelium are novel findings, expanding the number of possible agents of NGD and supporting the hypothesis on multicausal aetiology of this disease.     

Different immune response of pigs to Mycobacterium avium subsp. avium and Mycobacterium avium subsp. hominissuis infection

Mycobacterium avium subsp. avium (MAA) and Mycobacterium avium subsp. hominissuis (MAH) are the most common mycobacterial species isolated from granulomatous lesions in swine in countries with controlled bovine tuberculosis. This study is focused on the immunological aspect of MAA and MAH infection in pigs. We detected induction of humoral and cell-mediated immunity in experimentally infected pigs. Specific antibodies were analyzed in serum by ELISA and the IFN-γ release assay was used for evaluation of cell-mediated immunity. While MAA induced a significant increase of both types of immune responses, MAH-infected pigs had an unvarying level of specific antibodies and showed low cell-mediated immunity with high individual variability. The subsequent in vitro experiment confirmed the lower immunogenicity of the MAH strain in comparison to MAA. MAH-infected porcine monocyte-derived macrophages showed a weaker induction of pro-inflammatory mediators in comparison to MAA, which included mRNA for IL-1β, TNF-α, IL-23p19, IL-18 and chemokines CCL-3, CCL-5, CXCL-8 and CXCL-10. Additionally, qualitative proteomic analysis revealed 28 proteins exclusively in MAA and 7 proteins unique to MAH. In conclusion, closely related M. avium subspecies MAA and MAH showed different capacities to stimulate the porcine immune system. From a diagnostic point of view, the IFN-γ release assay showed higher sensitivity than the detection of specific antibodies by ELISA and seems to be an effective tool for discrimination of MAA-infected pigs. In the case of MAH infection, the IFN-γ release assay could fail because of the low immunogenic capacity of the MAH strain.     

Characterization of <Emphasis Type="Italic">Agrobacterium</Emphasis> species by capillary isoelectric focusing

Tumorigenic and non-tumorigenic strains of Agrobacterium tumefaciens, A. rhizogenes, A.rubi, and A.vitis were examined using capillary isoelectric focusing, phenotypic determinative tests, PCR and fatty acid analysis. The isoelectric points (pI) of the 40 strains investigated clearly differentiated the strains according to their respective species. The different species were characterized with the following pI values: A. tumefaciens 2.2, A. rhizogenes 4.0, A. rubi 2.15, and A. vitis 2.6. This differentiation corresponded to the phenotypic, PCR and fatty acid characterizations. Strains with the similar chromosomal background but different plasmid content, e.g. A.vitis strain S4, and F2/5 gave the same pI values. Strains of Rhizobium species differed from Agrobacterium strains in their pI values. The advantage of capillary isoelectric focusing over the phenotypic determinative tests, PCR and fatty acid analysis is its speed (15 min), relative simplicity, and the very small amount of chemicals used. This rapid and simple method is a major improvement over the classical methods of separation of Agrobacterium species and should prove useful for rapid characterization of Agrobacterium-like colonies isolated from plant tumours for epidemiological and generic diversity studies.     

Functional Properties of Dunaliella salina and Its Positive Effect on Probiotics

The unicellular green microalga Dunaliella is a potential source of a wide range of nutritionally important compounds applicable to the food industry. The aim of this study was to assess the effect of Dunaliella salina dried biomass on the growth and adherence of 10 strains of Lactobacillus, Lacticaseibacillus, and Bifidobacterium. The immunomodulatory, antioxidant, and cytotoxic effects of D. salina on human peripheral mononuclear cells and simulated intestinal epithelial cell lines Caco-2 and HT-29 were evaluated. Furthermore, the hypocholesterolemic effects of the microalgae on lipid metabolism in rats fed a high-fat diet were analyzed. The addition of D. salina biomass had a positive effect on the growth of nine out of 10 probiotics and promoted the adherence of three bifidobacteria strains to human cell lines. The antioxidant and immunomodulatory properties of D. salina were concentration-dependent. The inflammatory cytokines (TNF-α and IL-6) were significantly increased following Dunaliella stimulation at the lowest concentration (0.5% w/v). Eight week supplementation of D. salina to the diet of hypercholesteromic rats significantly decreased the serum concentrations of LDL-C, VLDL, IDL-B, and IDL-C. D. salina is not cytotoxic in intestinal cell models; it promotes adherence of selected bifidobacteria, it affords immunomodulatory and antioxidant effects, and its addition to diets may help decrease atherosclerosis risk factors.     

Combining ability under drought stress relative to SSR diversity in common wheat

Eight-parental diallel cross and SSR molecular markers were used to determine the combining ability of common wheat lines grown under well-watered (WW) and water-stress (WS) conditions. Analysis of variance of yield indicated highly significant differences among the progenies. General combining ability (GCA) determined most of the differences among the crosses. Specific combing ability (SCA) was also significant but less important. The estimates of GCA effects indicated that one line was the best general combiner for grain yield under drought. Neis genetic distance, measured using SSR markers, differed from 0.20 to 0.48 among the eight genotypes. The correlation of Neis genetic distance with SCA for grain yield and heterosis ranged from 0.4 to 0.5. These results indicate that the level of SCA and heterosis depends on the level of genetic diversity between the wheat genotypes examined. Microsatellite markers were effective in predicting the mean and the variance of SCA in various cultivars combinations. However, selection of crosses solely on microsatellite data would miss superior combinations.