Endocrine changes associated with overripening of ovulated eggs in goldfish, Carassius auratus L.

Changes in steroid hormone levels in the serum and ovarian fluid were studied during overripening in goldfish. Ovulated eggs retained in the ovarian cavity become overripe at around 12 h after ovulation and completely overripe 24 h after. Blood and ovarian fluid were taken at 0, 3, 6, 12, 18, and 24 h after ovulation. Estradiol-17ß (E 2) , testosterone (T), progesterone (P) and 17α,20ß-dihydroxy-4-pregnen-3-one (17α,20ß-P) in the serum were extracted directly with a solvent while those in the ovarian fluid were separated by HPLC before radioimmunoassay. Both serum and ovarian fluid P showed a highly significant decline at 18 h with a further decline at 24 h; P levels were higher in the ovarian fluid. Serum 17α,20ß-P showed a progressive and more rapid decline, decreasing significantly at 12 h with further decreases at 18 h and 24 h; the level was five-fold lower at 24 h compared to the 0 h level. Serum T increased significantly at 3 h which was maintained until 18 h, when it declined to 0 h level. No significant changes in E2 were observed in the serum, except for a significant difference between 6 and 24 h. There were no significant changes in E2, T and 17α,20ß-P in the ovarian fluid. Of the four steroids measured, only 17α,20ß-P and P showed changes which bear some correlation with the time course of overripening. The declines in the mean ratios of 17α,20ß-P/E2 in the serum and P/E2 in the ovarian fluid also appeared to have a good correlation with the time course of overripening. The postovulatory follicles (POFs) showed degenerative features which likewise correspond to the decline in P and 17α,20ß-P

OVERRIPENING OF OVULATED EGGS IN GOLDFISH, CARASSIUS AURATUS L

Ph.DDOCTOR OF PHILOSOPH

Cloning of mangrove red snapper (Lutjanus argentimaculatus) growth hormone cDNA and mRNA expression during early development

Growth hormone regulates growth and development in vertebrates. As a first step to understand the role of growth hormone in the regulation of growth and development of the mangrove red snapper Lutjanus argentimaculatus, the red snapper growth hormone (sGH) cDNA was cloned using reverse transcription - polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The expression of sGH mRNA in embryos and larvae was examined also by RT-PCR. Excluding the poly-A tail, the full-length red snapper GH cDNA is 945 base pairs (bp) long. It contains untranslated regions of 99 bp and 234 bp in the 5’ and 3’ ends, respectively. It has an open reading frame of 612 bp coding for a signal peptide of 17 amino acids and a mature hormone of 187 amino acid residues. Red snapper GH contains 4 cysteine residues and the typical polyadenylation site 16 bp upstream of the poly-A tail. Based on the amino acid sequence of the mature hormone, sGH shows higher sequence identity (>75%) to GHs of perciforms like grouper, seabass, tilapia and rabbitfish than to GHs of salmonids and carps. Semi-quantitative RT-PCR showed that expression of sGH mRNA commenced two days after hatching

Antibacterial properties of the microalgae Chaetoceros calcitrans

The antibacterial properties of the microalgae Chaetoceros calcitrans were assessed. Samples of C. calcitrans were first extracted in methanol, and then in different organic solvents of increasing polarity, n-hexane (n-Hex), dichloromethane (DCM) and ethyl acetate (EA) by liquid-liquid extraction. Solvent extracts were screened for antibacterial activity against four species of bacteria: Gram positive, Staphylococcus aureus and Bacillus subtilis; and Gram negative, Escherichia coli and Vibrio harveyi, with Amoxicillin as positive control, N-Hex extract, with significantly lower antibacterial activity than Amoxicillin, showed significantly higher activity than DCM and EA extracts, and least in methanolic extract. High antibacterial activity of n-Hex extract against all the microorganisms indicates that the bioactive components could be non-polar since the activity decreased as the solvent became more polar like methanol, and finally lost in aqueous extract. Results also showed that the extracts have a broad spectrum activity. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of all solvent extracts on all microorganisms tested ranged from 125 to 500 μg.mL-1. Partial purification and characterisation of the extracts confirmed the antibacterial activity in the non-polar fraction, which could be terpenes. The results suggest a good prospect in using C. calcitrans against Vibrio and other bacterial species.We thank the University of the Philippines (UP) System Creative and Research Scholarship Program and UP Visayas In-house Research Fund for funding the project, and SEAFDEC-AQD, Tigbauan, Iloilo, Philippines for the use of their facilities