Zebrafish (Danio rerio) as an animal model for bath infection by Flavobacterium psychrophilum

Flavobacterium psychrophilum is the causative agent of bacterial cold-water disease and rainbow trout syndrome in freshwater salmonid fish worldwide, generating injuries and high mortality rates. Despite several studies on this bacterium, the infection mechanism remains unknown due to limitations in the employed animal models. In this work, we propose using zebrafish (Danio rerio) as a model for studying bacterial pathogenicity. To substantiate this proposal, zebrafish infection by F. psychrophilum strain JIP 02/86 was characterized. Zebrafish larvae were infected using the bath method, and morphological changes and innate immune system activation were monitored using transgenic fish. Salmonid-like infection phenotypes were observed in 4.74% of treated larvae, as manifested by fin, muscle and caudal peduncle damage. Symptomatic and dead larvae accounted for 1.35% of all challenged larvae. Interestingly, infected larvae with no infection phenotypes showed stronger innate immune system activation than specimens with phenotypes. A failure of function assay for myeloid factor pu.1 resulted in more infected larvae (up to 43.5%), suggesting that low infection rates by F. psychrophilum would be due to the protective actions of the innate immune system against this bacterium in zebrafish larvae. Our results support the use of zebrafish as an infection model for studying F. psychrophilum. Furthermore, the percentage of infected fish can be modulated by disturbing, to varying extents, the differentiation of myeloid cells. Using this evidence as a starting point, different aspects of the infection mechanism of F. psychrophilum could be studied in vivo.     

Fortification of milk with calcium: effect on calcium bioavailability and interactions with iron and zinc

Calcium solubility, dialysability, and transport and uptake (retention + transport) by Caco-2 cells as indicators of calcium bioavailability have been estimated in the in vitro gastrointestinal digests of milk and calcium fortified milk. A significant linear correlation (p < 0.05) was obtained between calcium uptake and the amount of soluble calcium added to the cells, and also between percentage calcium uptake and the calcium measured in the analyzed samples. The solubility, dialysis, transport, and uptake values are higher (p < 0.05) for calcium fortified milks than for nonfortified milks; that is, calcium fortification increases not only calcium content but also its bioavailability. An inhibitory effect of calcium from fortified milks upon iron absorption was found. The observed effect of calcium from fortified milks upon zinc bioavailability depends on the in vitro method used, zinc solubility and dialysis decrease in calcium fortified milks, and percentage zinc uptake remains unchanged.     

Bioavailability of inorganic arsenic in cooked rice: practical aspects for human health risk assessments

Arsenic is present in rice grain mainly as inorganic arsenic. Little is known about the effect of cooking on inorganic arsenic content in rice and its bioavailability. This study evaluated total arsenic and inorganic arsenic in rice cooked with arsenic-contaminated water, the bioaccessibility of As(III) and As(V) after simulated gastrointestinal digestion, and the extent of arsenic retention and transport by Caco-2 cells used as a model of intestinal epithelia. After cooking, inorganic arsenic contents increase significantly. After simulated gastrointestinal digestion, the bioaccessibility of inorganic arsenic reached 63-99%; As(V) was the main species found. In Caco-2 cells, arsenic retention, transport, and total uptake (retention + transport) varied between 0.6 and 6.4, 3.3 and 11.4, and 3.9 and 17.8%, respectively. These results show that in arsenic endemic areas with subsistence rice diets, the contribution of inorganic arsenic from cooked rice should be considered in assessments of arsenic health risk.     

Chicken anemia virus and fowl adenoviruses: association to induce the inclusion body hepatitis/ hydropericardium syndrome

The effects of a simultaneous and/or a subsequent coinfection with chicken anemia virus (CAV) isolate 10343 and fowl adenovirus (FAV) isolate 341 in specific-pathogen-free light chickens were evaluated. The simultaneous coinfection was conducted by the intramuscular route, whereas the subsequent coinfection trial considered FAVs administered orally. In trial 1, 20-day-old chickens simultaneously coinfected with CAV (10343) and FAV (341) intramuscularly (i.m.) showed 55% mortality and characteristic signs and lesions of inclusion body hepatitis/hydropericardium (IBH/HPS). In contrast, birds singly infected with FAV i.m. showed 10% mortality due to IBH/HPS. Trial 2 showed that birds receiving FAV 341 orally at day 7 post-CAV intramuscular infection (group A) developed a mild form of IBH/HPS with presence of inclusion bodies (INIBs) in 60% of the group and virus-neutralizing antibodies against FAV 341. Group B (FAV orally 14 days after CAV) showed significant decreased weight gain, nonspecific microscopic lesions in the liver, spleen, bursa, and thymus, and an antibody response against FAV 341. However, no INIBs could be detected in the hepatocytes of these chickens. Group C (FAV orally 35 days after CAV) showed nonspecific histopathologic changes in the liver and no antibody response to FAV. The oral single infection with FAV isolate 341 induced neither mortality nor macroscopic lesions of IBH/HPS in the birds. The present results corroborate previous reports on pathogenicity of Chilean FAV isolates, which suggest that synergism with other viruses or prior immunosuppression is necessary to produce IBH/HPS in chickens. These results also suggest that the susceptibility of chickens to FAV oral infection resulting in IBH/HPS varies throughout the course of CAV infection.     

Ammonia reduction in seawater by Yucca schidigera extract: efficacy analysis and empirical modelling

Yucca schidigera is a plant native to southwestern USA and Mexico. Its extract has been used in the livestock industry to control ammonia accumulation in animal holding facilities, and to reduce ammonia concentration in animal excreta. This study investigated the potential and effectiveness of Y. schidigera extract (YUPE) for ammonia reduction in seawater. A dose–response experiment was conducted to determine the effect of different concentrations of YUPE at 0, 18, 36, 72 and 108 mg L⁻¹ on total ammonia nitrogen (TAN) at 1, 3 and 9 mg L⁻¹. At a higher YUPE dosage rate, higher TAN reduction was observed, and TAN reduction was highest during the first 12 h, and decreased thereafter. A stepwise multiple linear regression that included the initial TAN, YUPE concentration and time was developed, which accurately predicted empirical TAN concentrations. Applications of this model for ammonia management strategies were formulated for hypothetical tiger prawn (Penaeus monodon) rearing conditions. YUPE's efficacy for ammonia reduction, natural origin and safety make YUPE a potentially suitable compound for water quality management in mariculture.     

Live attenuated malaria vaccine designed to protect through hepatic CD8⁺ T cell immunity

Our goal is to develop a vaccine that sustainably prevents Plasmodium falciparum (Pf) malaria in ≥80% of recipients. Pf sporozoites (PfSPZ) administered by mosquito bites are the only immunogens shown to induce such protection in humans. Such protection is thought to be mediated by CD8(+) T cells in the liver that secrete interferon-γ (IFN-γ). We report that purified irradiated PfSPZ administered to 80 volunteers by needle inoculation in the skin was safe, but suboptimally immunogenic and protective. Animal studies demonstrated that intravenous immunization was critical for inducing a high frequency of PfSPZ-specific CD8(+), IFN-γ-producing T cells in the liver (nonhuman primates, mice) and conferring protection (mice). Our results suggest that intravenous administration of this vaccine will lead to the prevention of infection with Pf malaria.     

Arsenosugars in raw and cooked edible seaweed: characterization and bioaccessibility

The aim of this study was to examine arsenic species contents in raw and cooked edible seaweed and the bioaccessibility (maximum soluble concentration in gastrointestinal medium) of arsenosugars (glycerol ribose, phosphate ribose, sulfonate ribose, and sulfate ribose). For the analysis, a new chromatographic separation was developed in anion exchange, coupled with thermooxidation-hydride generation-atomic fluorescence spectrometry. An in vitro digestion (pepsin, pH 2; pancreatin-bile extract, pH 7) was applied to estimate arsenosugar bioaccessibility. Cooking of Undaria pinnatifida and Porphyra sp. did not alter the arsenic species present in the methanol-water extract, but it produced a substantial increase (2 and 5 times) in the As(V) extracted from Hizikia fusiforme. In all of the seaweeds analyzed, arsenosugar bioaccessibility was high (>80%) and did not vary as a result of cooking. Arsenosugar degradation as a result of in vitro digestion was not observed.     

Irbic acid, a dicaffeoylquinic acid derivative from Centella asiatica cell cultures

3,5-O-dicaffeoyl-4-O-malonilquinic acid (1) (irbic acid) has been isolated for the first time from cell cultures of Centella asiatica and till now it has never been reported to be present in the intact plant. Evidence of its structure was obtained by spectroscopic analyses (MS/NMR). Besides 1, cell cultures produce also the known 3,5-O-dicaffeoylquinic acid, chlorogenic acid, and the triferulic acid 2 (4-O-8′/4′-O-8″-didehydrotriferulic acid). Biological activities were evaluated for compound 1, which showed to have a strong radical scavenging capacity, together with a high inhibitory activity on collagenase. This suggests a possible utilization of this substance as a topical agent to reduce the skin ageing process.     

Physical,Compositional, and Wet‐Milling Characteristics of Mexican Blue Maize (Zea mays L.) Landrace

Mexico has the largest diversity of genetic resources for maize in the world, with about 59 different landraces. However, little is known about their wet‐milling characteristics. The aim of this study was to determine whether 15 Mexican blue maize (Zea mays L.) genotypes of Elotero de Sinaloa landrace collected in the northwestern region of Mexico have suitable wet‐milling properties. Great variability of physical, compositional, and wet‐milling characteristics among these blue maize genotypes was observed. The FAUAS‐457 and FAUAS‐488 maize genotypes had similar starch yield and starch recovery as reported for the wet‐milling industry, which indicated that they may be useful as a source of extractable starch. Residual protein levels in the starch fractions were in the range of 0.39–0.68%, and total solids recovery exhibited a mean value of 98.8%, indicating acceptable efficacy of the wet‐milling process. This process afforded starches from blue maize genotypes with low protein contents. Wet‐milling fractions correlated with the physical and chemical properties of the kernels. Our results indicate that Mexican blue maize genotypes contain characteristics that make them appropriate and utilizable at the industrial level, and they can also be valuable for improving wet‐milling characteristics of maize through breeding programs.