Time after seawater transfer influences immune cell abundance and responses to SAV3 infection in Atlantic salmon

Pancreas disease (PD) caused by salmonid alphavirus (SAV) severely affects salmonid aquaculture during the seawater phase. To characterize immune cells in target tissues for SAV infection, heart, pancreas and pyloric caeca were analysed from two groups of fish adapted to seawater for 2 and 9 weeks. The sections were scored for the relative abundance of cells expressing MHC class II, IgM, CD3, CD8 or neutrophil/granulocyte markers using immuno‐histochemical techniques. In general, necrosis of tissue was more severe in fish infected at 2 weeks post‐seawater transfer (wpt) compared with those infected at 9 wpt. At 9 wpt, there were higher numbers of MHC II⁺ cells in heart, pancreas and pyloric caeca, IgM⁺ cells in heart and pancreas, and CD3⁺ cells in pancreas compared to those infected at 2 wpt. The majority of the immune cells infiltrating PD‐affected tissues were MHC II⁺ and CD3⁺ cells suggesting that antigen‐presenting cells and T lymphocytes are the main types of immune cells responding to SAV infection. All the investigated cell types were also observed in pyloric caeca of infected fish, suggesting that this tissue may play a role in the immune response to SAV.     

Evaluation of the ability of altrenogest to control the equine estrous cycle

In our experience, altrenogest has not always been able to exert predictable control over the estrous cycle of the mare. Therefore, we examined 12 mares that were treated with altrenogest to identify reasons for its failure to control the estrous cycle. The mares were fed altrenogest for 15 to 20 days and were examined for follicle development, ovulation, and corpus luteum formation during treatment. Through the use of real-time ultrasonography and radioimmunoassay for progesterone, we concluded that altrenogest was unable to suppress the growth of follicles to preovulatory size in some mares, leading to ovulation during treatment or earlier than expected after the end of treatment. In addition, altrenogest did not appear to shorten the life-spans of the corpora lutea that were formed during treatment; in 4 mares, this resulted in the persistence of corpora lutea after the end of the suggested 15-day periods of treatment. The latter findings led us to suggest that if a luteolytic dose of prostaglandin had been given at the end of altrenogest treatment, there would have been improved control over the estrous cycle. The results of our study confirmed our clinical impressions that altrenogest may be satisfactory to control the equine estrous cycle under some circumstances, but it should not be used when precise control over ovulation is required.     

Effect of Untreated, Roasted and Germinated Black Gram (Phaseolus mungo) Flours on the Physico-chemical and Biscuit (Cookie) Making Characteristics of Soft Wheat Flour

Determination of the physico-chemical characteristics of composite soft wheat flours in which 5–25% (w/w) of the wheat flour was replaced with untreated, roasted and germinated black gram (Phaseolus mungo) flours (BGF) showed that when roasted BGF comprised 20% (w/w) of the blend, the increases in the ash and protein contents were 123% and 35%, respectively. The values for the gluten contents and the Zeleny and sodium dodecyl sulphate sedimentation test volumes for the composite flours indicated a weakening effect of BGF on the quality of soft wheat flour proteins, which could be beneficial for the preparation of biscuits (cookies). The alkaline water retention capacity values increased with the increasing addition of differently processed BGFs. Biscuit baking studies indicated that the diameter and spread ratio of biscuits were reduced, while the thickness increased, with increasing addition of all three BGFs, but the maximum reduction in diameter was observed with the addition of germinated BGF. The hardness value for biscuits increased with the addition of BGFs, but the effect was minimal with roasted BGF and maximal with germinated BGF. The surface grain score was reduced with increasing addition of BGF, but, in general, the roasted BGF showed the minimum adverse effect. From the overall biscuit making quality, addition of untreated BGF at the 15% level and of roasted and germinated BGFs at the 20% level were considered optimal for supplementing wheat flour.     

Permeability determination and pharmacokinetic study of nobiletin in rat plasma and brain by validated high-performance liquid chromatography method

In the present study, we are reporting permeability and pharmacokinetics of nobiletin in rat plasma and brain, using a validated reverse phase high performance liquid chromatographic method. Protein precipitation method was used for the extraction of nobiletin and coumarin (IS) from rat plasma and brain tissue. The system was run in isocratic mode with mobile phase consisting of potassium dihydrogen ortho-phosphate (pH 4.5; 0.04 mM) and acetonitrile in ratio of 50:50, v/v. The total chromatographic run time was 9.0 min. The method was proved to be accurate and precise at linearity range of 0.05–10 μg/mL with a correlation coefficient (r) of ≥ 0.994 in rat plasma and ≥ 0.995 in rat brain. The intra- and inter-day precision and accuracy values are found to be within the assay variability limits as per the FDA guidelines. Nobiletin was found stable in the battery of stability studies viz., bench-top, auto-sampler, freeze/thaw cycles and long term storage in a freezer at − 70 ± 10 °C. Maximum concentrations of nobiletin in both plasma and brain were observed at 1 h after single oral dosing (50 mg/kg). The maximum concentration in plasma and brain were 1.78 and 4.20 μg/mL, respectively. The AUC_0–t in plasma and brain were 7.49 and 20.66 μg·h/mL, respectively. The mean elimination half life (t_1/2) in plasma and brain were 1.80 and 11.42 h, respectively. The Parallel Artificial Membrane Permeability Assay (PAMPA) permeability of nobiletin was found to be high at both pH 4.0 and 7.0.     

Characterization and application of recombinant β-glucosidase (BglH) from <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> KCTC 1918

β-Glucosidase (β-1,4-D-glucoside glucohydrolase: EC.3.2.1.21) catalyzes the hydrolysis of β-glucosidic bonds between saccharides and aryl or alkyl groups. A gene encoding β-glucosidase from Bacillus licheniformis KCTC 1918, an anaerobic spore-forming soil bacterium, was cloned and characterized. The structural gene for the β-glucosidase consists of 1410 bp encoding 469 amino acid residues, and has a molecular weight of 53.4 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis with 12% separating gel. The enzyme activity was determined against pNPG as a substrate. The enzyme was optimally active at pH 6.0 (citrate-phosphate buffer) and 47°C. β-Glucosidase retained 100% of its original activity for 24 h. The activity of the enzyme was stimulated by glycerol and urea and was decreased by Ca²⁺, Cu²⁺, Hg²⁺, Mg²⁺, and Mn²⁺. In particular, Cu²⁺ had the strongest negative effect on β-glucosidase activity. The purified β-glucosidase was active against pNPG and cellobiose. When the β-glucosidase was tested for cellulose hydrolysis, the supplement of β-glucosidase with cellulose increased the glucose yield from pine wood powder by 139.8%.     

Tissue culture micropropagation of Douglas-fir

A procedure was developed for plantlet production from embryos of Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco]. The Medium for Conifer Morphogenesis, used at full strength, and supplemented with 10 M benzyladenine for 17 days produced an average of 6.8 shoots on more than 90% of the embryos. The percentage of shoot-forming embryos as well as the average number of shoots per embryo varied significantly among eight seedlots. For secondary multiplication, 89% of the adventitious shoots produced axillary buds on MCM with 5 M benzyladenine. However, 0.5 M BA was more suitable for the elongation of axillaries. Rooting ranged from 0–87% depending upon the treatment. The highest percentage was obtained with a 7-week incubation in peat:perlite containing 1/5-strength medium, 1% sucrose and 2.7 M naphthaleneacetic acid, followed by 5 weeks on peat:perlite with 1/5-strength major and minor salts, full iron and organics, 1% sucrose and 0.1% charcoal.