The Anti-Inflammatory Effect of Algae-Derived Lipid Extracts on Lipopolysaccharide (LPS)-Stimulated Human THP-1 Macrophages

Algae contain a number of anti-inflammatory bioactive compounds such as omega-3 polyunsaturated fatty acids (n-3 PUFA) and chlorophyll a, hence as dietary ingredients, their extracts may be effective in chronic inflammation-linked metabolic diseases such as cardiovascular disease. In this study, anti-inflammatory potential of lipid extracts from three red seaweeds (Porphyra dioica, Palmaria palmata and Chondrus crispus) and one microalga (Pavlova lutheri) were assessed in lipopolysaccharide (LPS)-stimulated human THP-1 macrophages. Extracts contained 34%–42% total fatty acids as n-3 PUFA and 5%–7% crude extract as pigments, including chlorophyll a, β-carotene and fucoxanthin. Pretreatment of the THP-1 cells with lipid extract from P. palmata inhibited production of the pro-inflammatory cytokines interleukin (IL)-6 (p < 0.05) and IL-8 (p < 0.05) while that of P. lutheri inhibited IL-6 (p < 0.01) production. Quantitative gene expression analysis of a panel of 92 genes linked to inflammatory signaling pathway revealed down-regulation of the expression of 14 pro-inflammatory genes (TLR1, TLR2, TLR4, TLR8, TRAF5, TRAF6, TNFSF18, IL6R, IL23, CCR1, CCR4, CCL17, STAT3, MAP3K1) by the lipid extracts. The lipid extracts effectively inhibited the LPS-induced pro-inflammatory signaling pathways mediated via toll-like receptors, chemokines and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling molecules. These results suggest that lipid extracts from P. lutheri, P. palmata, P. dioica and C. crispus can inhibit LPS-induced inflammatory pathways in human macrophages. Therefore, algal lipid extracts should be further explored as anti-inflammatory ingredients for chronic inflammation-linked metabolic diseases.     

T-2 toxin induced Salmonella Typhimurium intoxication results in decreased Salmonella numbers in the cecum contents of pigs, despite marked effects on Salmonella-host cell interactions

ABSTRACT: The mycotoxin T-2 toxin and Salmonella Typhimurium infections pose a significant threat to human and animal health. Interactions between both agents may result in a different outcome of the infection. Therefore, the aim of the presented study was to investigate the effects of low and relevant concentrations of T-2 toxin on the course of a Salmonella Typhimurium infection in pigs. We showed that the presence of 15 and 83 μg T-2 toxin per kg feed significantly decreased the amount of Salmonella Typhimurium bacteria present in the cecum contents, and a tendency to a reduced colonization of the jejunum, ileum, cecum, colon and colon contents was noticed. In vitro, proteomic analysis of porcine enterocytes revealed that a very low concentration of T-2 toxin (5 ng/mL) affects the protein expression of mitochondrial, endoplasmatic reticulum and cytoskeleton associated proteins, proteins involved in protein synthesis and folding, RNA synthesis, mitogen-activated protein kinase signaling and regulatory processes. Similarly low concentrations (1-100 ng/mL) promoted the susceptibility of porcine macrophages and intestinal epithelial cells to Salmonella Typhimurium invasion, in a SPI-1 independent manner. Furthermore, T-2 toxin (1-5 ng/mL) promoted the translocation of Salmonella Typhimurium over an intestinal porcine epithelial cell monolayer. Although these findings may seem in favour of Salmonella Typhimurium, microarray analysis showed that T-2 toxin (5 ng/mL) causes an intoxication of Salmonella Typhimurium, represented by a reduced motility and a downregulation of metabolic and Salmonella Pathogenicity Island 1 genes. This study demonstrates marked interactions of T-2 toxin with Salmonella Typhimurium pathogenesis, resulting in bacterial intoxication.     

O157:H7 and O104:H4 Vero/Shiga toxin-producing Escherichia coli outbreaks: respective role of cattle and humans

ABSTRACT: An enteroaggregative Verotoxin (Vtx)-producing Escherichia coli strain of serotype O104:H4 has recently been associated with an outbreak of haemolytic-uremic syndrome and bloody diarrhoea in humans mainly in Germany, but also in 14 other European countries, USA and Canada. This O104:H4 E. coli strain has often been described as an enterohaemorrhagic E. coli (EHEC), i.e. a Vtx-producing E. coli with attaching and effacing properties. Although both EHEC and the German O104:H4 E. coli strains indeed produce Vtx, they nevertheless differ in several other virulence traits, as well as in epidemiological characteristics. For instance, the primary sources and vehicles of typical EHEC infections in humans are ruminants, whereas no animal reservoir has been identified for enteroaggregative E. coli (EAggEC). The present article is introduced by a brief overview of the main characteristics of Vtx-producing E. coli and EAggEC. Thereafter, the O104:H4 E. coli outbreak is compared to typical EHEC outbreaks and the virulence factors and host specificity of EHEC and EAggEC are discussed. Finally, a renewed nomenclature of Vtx-producing E. coli is proposed to avoid more confusion in communication during future outbreaks and to replace the acronym EHEC that only refers to a clinical condition.     

Evaluation of beta3-adrenoceptor-mediated relaxation in intact and endotoxin-treated equine digital veins

OBJECTIVE: To investigate the functional expression of beta3-adrenoceptors (beta3-ARs) in equine digital veins (EDVs) and to examine whether beta3-AR relaxation was altered in EDVs incubated with endotoxin. SAMPLE POPULATION: Forelimbs obtained from 30 horses. PROCEDURE: Forelimbs were obtained from horses in an abattoir. Equine digital veins were carefully removed from distal portions of the forelimbs. Rings of dissected EDVs were mounted in 5-mL organ baths to record isometric tension in the presence of various beta3-AR agonists (SR 58611A, ZD 2079, and ZM 215001). RESULTS: In intact EDVs, isoprenaline, SR 58611A, ZD 2079, and ZM 215001 induced concentration-dependent relaxation. Isoprenaline and SR 58611A-induced relaxations were reduced or unaffected by nadolol, respectively. In intact EDVs, SR 58611A-induced relaxation was significantly reduced in the presence of 2 microM ZM 215001 (used as a beta3-AR antagonist). In endothelium-denuded EDVs or intact EDVs in the presence of a nitric oxide synthase inhibitor, isoprenaline and SR 58611A-induced relaxations were significantly decreased. The endothelium-independent relaxation to SR 58611A was significantly inhibited in the presence of ZM 215001. In endotoxin-treated EDV, isoprenaline- and SR 58611A-induced relaxations were significantly reduced. In these conditions, cycloheximide (a protein synthesis inhibitor) and ibuprofen (a cyclooxygenase inhibitor) restored the relaxant response to SR 58611A. CONCLUSIONS AND CLINICAL RELEVANCE: Beta3-adrenoceptors are functionally expressed in EDVs. Incubation in the presence of endotoxin, used as an in vitro model of laminitis, induced an alteration of beta-AR-mediated relaxations in EDVs, which could be the consequence of cyclooxygenase induction and subsequent prostanoid production.     

Recovery of Moraxella ovis from the bovine respiratory tract and differentiation of Moraxella species by tDNA-intergenic spacer PCR

The purpose of the present study was to identify Moraxella (M.)—like organisms recovered from calves suffering from respiratory disease down to species level by means of tDNA-intergenic spacer length polymorphism analysis (tDNA-PCR), and to perform antimicrobial susceptibility testing of these isolates using an agar dilution technique. A total of 16 isolates originating from 12 unrelated occasions were identified as Moraxella ovis, and tDNA fingerprinting showed clear delineation from other Moraxella species. The minimal inhibitory concentrations (in μg/mL) for 90% of the investigated isolates were ≤0.03 for ampicillin; 0.25 for ceftiofur; 0.5 for oxytetracycline; 8 for gentamicin; 64 for spectinomycin; 0.5/9.5 for the combination trimethoprim-sulfonamides; 4 for erythromycin; 8 for tilmicosin; 1 for florfenicol and 0.125 for enrofloxacin.     

Multiplex PCR assay for the detection of high virulence rabbit Staphylococcus aureus strains

High virulence rabbit Staphylococcus aureus strains, which are clonal in origin, are responsible for the spread of chronic staphylococcosis at the rabbit flock level. The aim of the present study was to develop a multiplex PCR assay that can be used for the identification of these high virulence strains. Two targets of the assay were the bbp and the selm genes, which have recently been shown to occur specifically in high virulence isolates. A third target was a sequence designated "flank", which was derived from a previously generated high virulence specific RAPD pattern. Furthermore, the femA gene, which is specific for S. aureus, was incorporated in order to avoid false negative results due to insufficient DNA preparation. The multiplex PCR was successful at differentiating the 26 typical high virulence and 50 low virulence rabbit S. aureus strains incorporated in the present study. Therefore it is useful for the initial screening of newly acquired breeding stock, in order to prevent the intake of high virulence strains in rabbitries.     

Pinpointing the role of Aeromonas salmonicida in the development of skin ulcerations in common dab (Limanda limanda)

Aeromonas salmonicida was isolated from ulcerations in common dab (Limanda limanda). An experiment was performed to pinpoint its role in ulceration development, considering the importance of the skin barrier and the pigmented and non-pigmented sides. The skin of dab was treated in three zones, one where scales and epidermis were removed, one where mucus was discarded and one non-treated zone. Fish were tagged to allow individual identification and challenged with A. salmonicida. Mortality and severity of the developing lesions were recorded for 21 days post-inoculation. Starting 12 days post-inoculation, mortality occurred gradually in challenged fish; however, no direct cause could be established. Both control fish and challenged fish developed ulcerations containing A. salmonicida. Sequencing of vapA gene revealed that isolates retrieved from both groups were distinct, suggesting the presence of A. salmonicida prior to the trial. Most ulcerations developed in zones where skin was removed, suggesting that abrasion might be a predisposing factor in ulceration development. Ulcerations were also observed at the insertion site of the tag, where exposed muscle tissue might have favoured the development of ulcerations. In conclusion, A. salmonicida seems to be involved in the development of skin ulcerations in dab, although the exact pathogenesis needs to be elucidated.     

Moisture content variability in kiln-dried <Emphasis Type="Italic">Gmelina arborea</Emphasis> wood: effect of radial position and anatomical features

Gmelina arborea is one of the most important species for plantation in tropical areas. However, high variability in final moisture content (MC_f) is a problem in the drying process. This study sought to determine the variability in MC_f in relation to distance from the pith (DP), and anatomic elements. Boards of 1.2 cm in thickness obtained from bark to the pith from 15-year-old plantation trees were dried in a pilot kiln. Anatomical characteristics, initial moisture content (MC_i), and specific gravity (SG) were also determined. Pearson correlation matrix was conducted between MC_f and the other variables. DP, fiber length, and ray width were correlated negatively with MC_f, while fiber diameter was positively correlated. Other studied anatomical properties showed no statistical correlations. Other wood properties such as SG and MC_i showed some influence on MC_f. DP can be considered as a good practical predictor for MC_f in G. arborea because some wood properties are related to MC_f. However, other wood characteristics that are not related with DP also influence MC_f, such as the presence of sapwood, MC_i variation between and within trees, fiber diameter, and ray width. These results suggest that MC_f variation in G. arborea is difficult to determine.     

Heritability of quantitative traits in segregating common bean families using a Bayesian approach

Genetic parameters for six quantitative traits in the early generation of segregating populations of common beans (Phaseolus vulgaris L.) were evaluated. A Bayesian approach was used for estimating the variance components, breeding values and broad sense heritability of the quantitative traits under analysis. The Markov Chain Monte Carlo method was utilized to analyze the contribution of genes affecting complex traits. Twenty-four F₃ families were evaluated in the field during 2005 in Santa Catarina, southern Brazil. With regard to the grain yield and yield components, the additive variances were relatively similar to the dominance variances. This result is confirmed by the 95% credible set from the posterior distribution. The mean estimates of broad-sense heritability (H²) varied from 11.5% to 64.2%. The heritability estimates of yield and yield components were higher than the estimates for the number of days until flowering and reproductive period. However, for grain yield, the 95% heritability credible set included the heritability estimates from point of crop duration. The predicted genetic gain reached the highest value for the number of pods per plant (10.95%). Days to flowering and reproductive period had the lowest values of genetic advance. One hundred seed-weight, grain yield and seeds per pod exhibited a similar predictable level of genetic gain: GA = 5.73%, 5.81% and 4.77%, respectively. The Bayesian framework provided information that is useful for a breeding program, since it contributes to the understanding of how quantitative traits are genetically controlled.     

Distribution patterns and population structure of the blue shark (Prionace glauca) in the Atlantic and Indian Oceans

The blue shark (Prionace glauca) is the most frequently captured shark in pelagic oceanic fisheries, especially pelagic longlines targeting swordfish and/or tunas. As part of cooperative scientific efforts for fisheries and biological data collection, information from fishery observers, scientific projects and surveys, and from recreational fisheries from several nations in the Atlantic and Indian Oceans was compiled. Data sets included information on location, size and sex, in a total of 478,220 blue shark records collected between 1966 and 2014. Sizes ranged from 36 to 394 cm fork length. Considerable variability was observed in the size distribution by region and season in both oceans. Larger blue sharks tend to occur in equatorial and tropical regions, and smaller specimens in higher latitudes in temperate waters. Differences in sex ratios were also detected spatially and seasonally. Nursery areas in the Atlantic seem to occur in the temperate south‐east off South Africa and Namibia, in the south‐west off southern Brazil and Uruguay, and in the north‐east off the Iberian Peninsula and the Azores. Parturition may occur in the tropical north‐east off West Africa. In the Indian Ocean, nursery areas also seem to occur in temperate waters, especially in the south‐west Indian Ocean off South Africa, and in the south‐east off south‐western Australia. The distributional patterns presented in this study provide a better understanding of how blue sharks segregate by size and sex, spatially and temporally, and improve the scientific advice to help adopt more informed and efficient management and conservation measures for this cosmopolitan species.