Detection of prostaglandin E<Subscript>2</Subscript> in polychaete <Emphasis Type="Italic">Perinereis</Emphasis> sp. and its effect on <Emphasis Type="Italic">Penaeus monodon</Emphasis> oocyte development in vitro

Prostaglandins are involved in the reproductive processes in a variety of animals, including crustaceans. It was found that polychaetes, the best maturation diet for shrimp broodstock, possessed the greatest variation of prostaglandin E₂ (PGE₂) when compared with other live feeds. The level of PGE₂ varied according to sizes, feed intake, sources and type of polychaete. The matured and also larger sand polychaete Perinereis sp. contained higher PGE₂ levels than younger and smaller sand polychaetes (18.16 ± 5.82 ng PGE₂ mg⁻¹ protein for polychaetes at an average length of 10 cm up to 160.8 ± 37.09 ng PGE₂ mg⁻¹ protein for polychaetes at an average length of 17 cm). The PGE₂ levels in ovaries and haemolymph of female shrimp fluctuated with the developmental stage of the ovaries. The highest concentration of PGE₂ in haemolymph was at stage 3 of ovarian development, whereas the highest concentration of PGE₂ in shrimp ovaries was at stage 4. In vitro incubation of Penaeus monodon pre-vitellogenic oocytes with polychaete extract and synthetic PGE₂ demonstrated that both PGE₂s enhanced oocyte development, especially during late development and ovulation. The putative role of PGE₂ from polychaetes or the presence of PGE₂ in polychaetes may be a factor in their role as a dietary constituent required for shrimp oocyte development.     

The effects of ozone and probiotics on the survival of black tiger shrimp (Penaeus monodon)

Total ozone production (TOP) from an ozonator, residual ozone concentration (ROC) in water, and the effects of ozone with or without probiotic supplemented feeds on bacterial growth, and shrimp (Penaeus monodon) survival were investigated. Minimal effective ROC to inhibit 3 log units of Vibrio harveyi D331 for 6 h and 2 log units of Bacillus S11 for 9 h was 0.38 g O₃/l of ROC from 5-min ozonation. Shrimp postlarvae exposed to 0.34–0.50 mg O₃/l ROC (8-h ozonation) caused loss of balance, immobility and destruction of gill lamellar epithelium. In vivo treatment of juvenile P. monodon reared on probiotic feed for 1 month revealed that 0.35 mg O₃/l ROC (30-min ozonation) effectively inhibited 3 log units of V. harveyi D331 for 24 h. At this ROC dosage, there was no effect either on shrimp or on intestinal probiotics (Bacillus S11). Shrimp survival from probiotic treatment, coupling with ozonation, increased significantly (P≤0.05) compared with controls.     

Maturation diet based on fatty acid content for male Penaeus monodon (Fabricius) broodstock

The contents of three essential fatty acids, arachidonic acid (AA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), from wild Penaeus monodon broodstock were evaluated in comparison with natural diet fed P. monodon. Spermatophores of wild male broodstock contained higher levels of AA than those of artificial diet fed males. Polychaetes had higher proportion of AA to EPA and DHA at 5.8:5.5:1 in mud polychaetes followed by 12:7:1 in sand polychaetes, while DHA was a preferential n‐3 highly unsaturated fatty acid (HUFA) in squids and fish. The experimental feed was constructed to simulate the HUFA profile of polychaetes (AA:EPA:DHA as 5:1:1) and then fed to farmed male black tiger prawn broodstock for 1 month. The results exhibited comparable reproductive characteristics to wild male suggesting the possibility of replacing wild males with pond‐reared males. Rearing farmed males in a test unit for a month did not reduce the quality of prawn sperm. Reproductive performance indices (sperm sac weight, total number of sperm, percentage of live sperm, percentage of abnormal sperm) from the males of all treatments were not statistically different except in males fed with pellets. Control (live feeds) and combined diet provided better reproductive performance in pond‐reared males. Analysis of AA, EPA and DHA in reproductive tissues, hepatopancreas and muscle of treated animals in each treatment revealed an accumulation of dietary HUFA into reproductive tissues. No evidence of transfer of HUFA from hepatopancreas or muscle to spermatophore was found.     

Identification of progesterone and 17α-hydroxyprogesterone in polychaetes (Perinereis sp.) and the effects of hormone extracts on penaeid oocyte development in vitro

A possible factor behind the successful use of polychaetes for prawn broodstock diet was investigated. Some steroid hormones in polychaetes have been suspected as being a potential component for prawn reproductive system development. Progesterone (P4) and 17-alpha hydroxyprogesterone (17α-OHP4) were extracted from polychaetes (Perinereis sp.) and female Penaeus monodon broodstock at various maturation stages. Concentration of both hormones was determined using RIA or HPLC. Immature natural polychaetes (2 months old) displayed the highest level of P4 and 17α-OHP4 compared to more mature ones (4, 6 and 8 months old polychaetes) (47.48 ± 6.70 ng P4/mg Protein and 143.74 ± 28.70 ng 17α-OHP4/mg Protein) whereas opposite results were received with cultured polychaetes in which a reverse relationship between P4 and its derivative with polychaete maturation was obtained. Diverse concentrations of both hormones may be linked with other factors such as types of polychaetes, habitats, seasons, and feed intakes. Twenty-four hours in vitro incubation of prawn previtellogenic oocytes with P4 or 17α-OHP4 extracted from polychaetes or with synthetic hormones, significantly increased percentages of vitellogenic oocytes and oocytes with cortical rod compared to the control with no hormones. P4 was more effective in enhancing the final maturation of oocytes while 17α-OHP4 had more effects on vitellogenic oocytes. Synthetic steroid hormones at equal hormone concentrations produced similar results to steroid hormones extracted from natural polychaetes.