Expression of reproduction and antioxidant-related genes in crayfish Cherax quadricarinatus fed with dietary feed additives

We evaluated the impact of a combined vitamin E (vit E), a mixture of nucleotide (mixed-NT), Haematococcus pluvialis (H. pluvialis), arachidonic acid (AA), and yeast extract powder (YEP) supplementations on the relative mRNA expression of reproduction and antioxidant-related genes in red claw crayfish Cherax quadricarinatus using an orthogonal design (L16 4⁵) plus control diet for 70 days feeding trial. Five factors with four levels: A, vit E ^{40, 160, 240, and 320 mg/kg diet}; B, mixed-NT ^{2, 4, 6, and 8 g/kg diet}; C, H. pluvialis ^{2, 4, 6, and 8 g/kg diet}; D, AA ^{5, 10, 15, and 20 g/kg diet}; E, YEP ^{2.5, 5, 10, and 15 g/kg diet} were studied. The relative mRNA expression of reproduction-related genes (vitellogenin (vtg) and fatty acid-binding protein (fabp)), and antioxidant-related genes (heat shock protein 70 (hsp70), glutathione peroxidase (gpx), glutathione S-transferase (gst), mitochondrial manganese superoxide dismutase (mmnsod), and ferritin) in hemocyte, hepatopancreas, and ovary tissues were measured. There was a major depression in gene expression levels throughout the experimental diet at high vit E ^320 mg/kg level with other factors. Expression of antioxidant enzyme genes significantly increased in treatments of T9, T10, T11, and T12, while significantly decreasing in treatment of T13 and other treatment groups with high vit E ^320 mg/kg levels with the other factors. Gene expression of vtg and fabp demonstrated similar patterns to those of antioxidant genes. These results provide evidence that combined supplementations of vit E, mixed-NT, H. pluvialis, AA, and YEP with vit E ^240 mg/kg, mixed-NT ^8 g/kg, H. pluvialis ^4 g/kg, AA ^5 g/kg, and YEP ^10 g/kg may present a novel strategy for improving reproduction performance and antioxidant activities in RC-crayfish, C. quadricarinatus by the positive impact on the molecular level of reproduction and antioxidant genes.

     

Effect of traditional methods of processing on the electrophoretic patterns of faba bean water soluble proteins

A number of national dishes are prepared from the dry seeds of faba bean by soaking, germination (nabet), cooking (bisara and medammis) or frying a dough (falafel). Soaking (12 h) decreased the intensity of bands in the standard PAGE and increased the charge heterogeneity of neutral proteins in PAGIF. Germination (24 or 60 h) changed the patterns for esterases, peroxidases and amylases, but not for phosphorylases. The charge distribution of proteins was markedly changed in the basic region. Roasting did not affect the distribution in standard PAGE. All bands in the basic region of PAGIF disappeared. High MW-proteins wre degraded to low MW-proteins as seen in SDS-PAGE. The cooking broth and the cooked beans of nabet soup and medammis had more or less similar patterns in the standard PAGE. The broth of nabet soup had larger number of low MW-proteins than the bean extract in SDS-PAGE. Bisara proteins showed almost the same patterns as the corresponding proteins of the cooking broth of nabet soup. Falafel proteins showed patterns very similar to that obtained from roasted beans in the SDS-PAGE. PAGIF gave a different but characteristic charge distribution for falafel.     

Genotyping of kappa-casein gene in Egyptian buffalo bulls

The present work is undertaken mainly for genotyping of kappa-casein gene in buffalo bulls. DNA was extracted from the frozen semen of nine buffalo bulls used in artificial insemination in addition to 40 blood samples of male buffalo calves for identification and genotyping of kappa-casein gene by PCR-RFLP assay. Different restriction endonucleases, including HindIII, HinfI and Taq1 were used. The PCR product of the primer specific for K2 and KY gives the specific band at size 379 and 280 bp respectively. Digestion of 379 bp fragment K2 by restriction endonuclease HindIII generated two fragments of 156 and 223 bp. For fragment 280 bp kappa-casein KY, the above mentioned endonuclease generated two fragments of 180 and 100 bp. Digestion of the 379 bp PCR product with HinfI resulted two restriction fragments of 288 and 91 bp. On the other hand, the amplified DNA (379 bp) from buffalo remained undigested by Taq1 restriction enzyme.In conclusion, the PCR-RFLP technique demonstrated that bulls were monomorphic for the kappa-casein gene; they possess only allele B in homozygosis form.