Effects of different feeding regimes on ovarian maturation and spawning of pond-reared giant tiger prawn in Thailand

Large pond-reared Penaeus monodon were used in three maturation and spawning trials lasting 60 days. One group of prawns was fed a fresh diet only; a second group was supplied a fresh diet combined with a formulated pelleted diet; a third group of prawns received the formulated diet only. Total maturation (N = 66 and 55) and spawning events (N = 63 and 48) were much greater for prawns receiving only the fresh diet, and the combined fresh with formulated diets, respectively. Prawns receiving only the formulated diet matured only 12 times, and spawned 11 times. Prawns fed a fresh diet produced significantly more eggs than those given a formulated diet alone. No significant differences were seen in eggs produced per spawning event, % fertilization, % hatch or % metamorphosis from egg to protozoea. Female prawns exhibited greater mortality than did males, while females on the fresh diet only had greater mortality than did other females, presumably due to greater handling and other stress factors. Female prawns on a combined diet had lower mortality than other females. The formulated diet used is suitable for supplementation, but not as a sole maturation food.     

Sperm Quality of Pond-Reared and Wild-Caught Penaeus monodon in Thailand1

Sperm quality, as determined by visual examination and by reaction with “egg-water” was not significantly different (P > 0.05) for sperm obtained by electro-ejaculation from ablated or non-ablated pond-reared Penaeus monodon. Nor was sperm quality different between pond-reared and wild-caught prawns. Normal sperm, determined by appearance, ranged from 17.1 to 21.0%, while reactive sperm ranged from 1.5 to 3.0%. There were, however, significant correlations (P < 0.01) between spermatophore weight and prawn weight (r= 0.73, N= 434). Male prawns weighing 4150 g produced on average spermatophores weighing 22.7 mg and containing 0.8 million sperm, while prawns weighing 61-90 g produced on average spermatophores weighing 56.6 mg with 2.5 million sperm. Ablation did not increase spermatophore size or sperm quality, although it significantly increased mortality of ablated males. Male prawns could be re-ejaculated at about weekly intervals with no change in sperm quality. Wild-caught female prawns artificially inseminated with spermatophores from electro-ejaculated males produced normal spawns with 51% average egg fertilization, and 41% nauplii hatch success. Nauplii hatch success following spawning increased from >60% for newly inseminated females to near zero after 30 days post-insemination, indicating spermatophore depletion and/or deteriorated sperm quality during spermatophore storage in the thelycum. The findings of the present study indicate that electro-ejaculation and artificial insemination are relatively simple and practical methods for improving captive reproduction performance of closed-thelycum prawns such as P. monodon, and that pond-reared and wild-caught males produced sperm of similar quality.     

Serotonin stimulates ovarian maturation and spawning in the black tiger shrimp Penaeus monodon

Serotonin (5-hydroxytryptamine, 5HT) has been reported to induce ovarian maturation and spawning in the crayfish Procambarus clarkii and white Pacific shrimp Litopenaeus vannamei. The aim of this study was to explore the role of exogenous 5HT on the reproductive performance of the black tiger shrimp Penaeus monodon. 5HT solution was injected into domesticated P. monodon broodstock at 50 μg/g body weight and ovarian maturation and spawning were recorded. The presence of 5HT in the ovary and oviduct of P. monodon was also studied by immunohistochemistry and its levels in the ovary by enzyme link immunoabsorbance assay (ELISA). The 5HT-injected P. monodon developed ovarian maturation and spawning rate at the level comparable to that of unilateral eyestalk-ablated shrimp. Hatching rate and the amount of nauplii produced per spawner were also significantly higher in the 5HT-injected shrimp, compared to the eyestalk-ablated shrimp. 5HT-positive reactions were found in the follicular cells of pre-vitellogenic oocytes, in the cytoplasm of early vitellogenic oocytes and on the cell membrane and cytoplasm of late vitellogenic oocytes. 5HT in the ovary was present at 3.53 ± 0.26 ng/mg protein level in previtellogenic stage and increased to 17.03 ± 0.57 ng/mg protein level in the mature stage of the ovary. The results suggest a significant role of 5HT, possibly directly on the ovary and oviduct, on the reproductive function of female P. monodon.     

Maturation and spawning performance of pond-reared Penaeus merguiensis in different combinations of temperature, light intensity and photoperiod

The combined effects of temperature (23 and 27 °C), light intensity (1100 and 2 lux or ≈ 21.5 and 0.04 μEinst m^?2 s respectively) and photoperiod (10 h light/14 h dark and 14 h light/10 h dark) on ovarian maturation and spawning performance of ablated pond‐reared Penaeus merguiensis were investigated in a 51‐day experiment. The results showed that temperature was the most influential factor, followed by light intensity, whereas the effect of photoperiod was minor. As the maturation process progressed, the effects of temperature and light intensity became stronger while that of photoperiod became less significant. Temperature significantly affected all the reproductive parameters assessed. Prawns in the 27 °C treatments outperformed those in the 23 °C treatments (P < 0.05). The effect of light intensity was found to have a significant effect (P < 0.05) only on the proportion of spawners (prawns that spawned) and spawning rate. More prawns spawned at a higher rate under dim light (2 lux) than under strong light (1100 lux). Photoperiod affected only the proportion of prawns reaching stage III of ovarian development (P < 0.05). There were interactions between temperature and light intensity affecting the proportion of prawns reaching stage III and, although not significantly, the proportion of spawners (P = 0.177), fecundity (P = 0.134) and survival (P = 0.061). Overall, it is recommended that a combination of 27 °C, 2 lux and 10 h light/14 h dark is suitable for the maturation of pond‐reared P. merguiensis. There were indications that temperature can be used to control the rate of ovarian maturation. Also, it is possible to increase light intensity up to 1100 lux in P. merguiensis hatcheries if prawns are ablated.     

Effects of dietary mannan oligosaccharide on growth,survival, physiological,immunological and gut morphological conditions of black tiger prawn (Penaeus monodon Fabricius 1798)

The aim of this experiment was to determine the effects of dietary inclusion with mannan oligosaccharide (Bio‐Mos, Alltech, Nicholasville, KY, USA) on growth, survival, physiological and immunological conditions and gut morphology of the black tiger prawn (Penaeus monodon). Five diets supplemented with MOS at 0 g kg^?1 (control diet), 1, 2, 4 and 8 g kg^?1 were fed to the prawn juveniles (0.4 ± 0.06 g, total weight) for the duration of 63 days. Growth was the highest (P < 0.05) when the prawns were fed the 1 g kg^?1 MOS included diet. Wet tail muscle index (Tw/B), dry tail muscle index (Td/B) and tail muscle protein (Tmp) were higher (P < 0.05) in the prawns fed MOS included diets MOS compared with the prawns fed the control diet. Total haemocyte counts (THCs) of the prawns fed MOS included diets were higher (P < 0.05) than THCs of the prawns fed the control diet. Epithelium layer and epidermal cell density of the gut of the prawns fed 1 g kg^?1 and 2 g kg^?1 MOS diets were better than the prawns fed the control and other MOS diets. The results imply a positive effect of dietary supplementation of 1–2 g kg^?1 MOS in the culture of black tiger prawns.     

New Modified Dipping Method Using 4-hexylresorcinol for Preventing Blackspot Formation in Prawns

Industry acceptance of alternative sulphite treatments for preventing blackspot in prawns is limited. This study demonstrated that dipping prawns (Penaeus monodon and Fenneropenaeus merguiensis) in a less concentrated solution for longer times provides better control of blackspot formation than the recommended 2 minute dip in 50 mg/L 4-hexylresorcinol. Protection improved as modified dipping times and solution concentrations increased. Blackspot protection provided by most of the modified dip methods was more effective than a 1 minute dip in 1% sodium metabisulphite. The recommended 2 minute dip method should be limited to 125 kg when used on consecutive 25-kg batches of prawns. Yields increased by less than 10% even after 72-hour exposure in the modified dip. The new dipping method meets most international residue standards when applying the modified treatments evaluated.     

A study of the arachidonic acid requirements of the giant tiger prawn, Penaues monodon

This study examined the dietary requirement of arachidonic acid (ARA) when that of linoleic acid (LOA), the natural precursor to ARA, was also satisfied with linolenic acid (LNA) and also with and without the other key dietary highly unsaturated fatty acids (HUFA). Growth by prawns fed diets supplemented with ARA was poorer than in diets where it was not present. Supplementation of ARA to diets with either optimized HUFA or just optimised poly unsaurated fatty acids (PUFA) (i.e. LOA, LNA) resulted in poorer growth. Growth was poorest by prawns (215 ± 13%) fed diets with ARA supplemented at 20% of the total fatty acids but including 7% LOA, 21% LNA and 4% of both eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Growth was best in prawns fed diets devoid of ARA but with 7% LOA and 21% LNA (350 ± 19%). Prawns fed the reference diet (348 ± 21%) and the other diet devoid of ARA but containing about 7% LOA, 21% LNA and 4% of both EPA and DHA (345 ± 18%) had similar growth. The growth responses were not effects of altered lipid or fatty acid digestibilities. Indeed supplementation of ARA to the diet marginally improved the digestibility of the total neutral lipid in the diet and the digestibilities of some other dietary fatty acids. The amount of lipid in the digestive glands of prawns fed with the diets was reduced by the inclusion of ARA in the dietary lipids. Composition of the lipids in the digestive gland (DG) of the prawns was almost directly related to the composition of their dietary lipids. The proportion of ARA in the total fatty acids increased with level of supplementation of dietary ARA. An increased level of dietary ARA reduced the proportion of EPA, DHA in the DG lipid and also the total n‐3 and n‐6 fatty acids in the DG lipid. The results of this study support that addition of ARA to the diet of Penaues monodon when the other key essential fatty acids (EFA) have been optimized, does not improve their growth performance. It is suggested that key cause for this response may lie in the importance of the balance of the n‐3 to n‐6 fatty acids in the diet of these animals.     

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.     

Haemagglutination as a low-cost detection method for gill-associated virus and by inference, yellowhead virus in Penaeus monodon Fabricius, 1798

A quantitative, low‐cost test based on haemagglutination (HA) using chicken erythrocytes was developed to indicate the viral load of Australian yellowhead‐like virus, gill‐associated virus (GAV), in Penaeus monodon. The study tested the haemolymph, gill, lymphoid organ, heart, sub‐cutaneous tissue, eye stalk, pleopods, uropods and the central nerve cord for agglutination activity in 100 prawns, with the haemolymph and gill tissue giving the highest end‐point titres of 1:1370 and 1:361 respectively. The sensitivity of the test was demonstrated by testing two different populations of P. monodon, which showed a highly significant difference (P<0.001) in HA activity, indicating a difference in viral load. By testing three other penaeid prawn species (n=20), Penaeus esculentus, Penaeus merguiensis and Penaeus longistylis, and the crayfish Cherax quadricarinatus, it was demonstrated that natural agglutinins were not causing the high agglutination in the population of P. monodon being tested. It was also demonstrated that there was no effect of freezing and thawing of samples on HA activity. The speed and low cost of this test makes it a very useful tool, particularly in the developing world, for on‐farm testing of penaeid prawns to indicate yellowhead virus and GAV loads which can contribute to management practices with respect to harvesting of ponds.     

Histological examination of final oocyte maturation and atresia in wild and domesticated Penaeus monodon (Fabricius) broodstock

Oocyte maturation and gonadosomatic index (GSI) of eyestalk ablated Penaeus monodon females collected from the wild and from two first‐generation domesticated lines were assessed. Frequency and diameter of the different oocytes, and the intensity of oocyte atresia, were compared among groups through histological assessments of the sections of the middle ovarian lobe. Digitized images from ovary sections were used to record the frequency and diameter of different oocyte types. Spawning performance of the three groups were expressed in terms of the percentage of females that spawned at least once (productive females), time from eyestalk ablation to first spawning (latency period) and the number of spawnings per female stocked. Final ovarian maturation was attained in all groups, as indicated by the presence of mature oocytes with cortical rods (cortical oocytes), dark‐green ovarian colour and high GSI values (5.83–6.86%). However, domesticated groups presented significant larger immature oocyte types (previtellogenic and yolky oocytes) and smaller cortical oocytes compared with wild females, indicating a reduced vitellogenic activity during final oocyte maturation. Additionally, the frequency of atresia was comparatively higher for both domesticated groups, which could be related to their inferior spawning performance. The implications of these results on the reproductive potential and development of domesticated P. monodon are discussed.