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.     

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.     

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.     

Evaluations of lactic acid bacteria as probiotics for juvenile seabass Lates calcarifer

Lactic acid bacteria (LAB) were isolated from adult, wild‐caught and farmed seabass (Lates calcarifer) intestines for evaluation as possible probiotics using the well agar diffusion method. Five LAB isolates (designated as LAB‐1–5) were found to inhibit Aeromonas hydrophila, a known seabass pathogen. Median lethal concentrations (LC₅₀) of A. hydrophila on juvenile seabass were measured in aquaria. Median lethal concentration values of 7.76, 7.47 and 7.26 log₁₀ CFU mL^?1 for 72, 96 and 120 h, respectively, were found. Juvenile seabass (0.6±0.2 g) were cultured in aquaria and fed individual LAB‐1–5 fortified feeds with 7 log₁₀ CFU g^?1 LAB. Seabass fed LAB‐4 fortified feed had significantly greater growth (P<0.05) than fish fed other feeds. Seabass fed LAB‐4 also had greater survival, but this was non‐significant (P<0.05). Challenge tests of LAB‐4 fed seabass with A. hydrophila at ～7 log₁₀CFU mL^?1 yielded significantly greater survival compared with control seabass (P<0.05). Aeromonas hydrophila infections in seabass were confirmed by observing disease manifestation and by immunohistochemistry techniques. LAB‐4 was preliminarily identified using lactic acid analysis, biochemical and physical characteristics. It was further identified using 16S rDNA sequencing. LAB‐4 was identified as Weissella confusa (identity of 99%). GenBank accession number for the 16S rDNA sequence for LAB‐4 was AB023241.     

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.     

Effects of Broodstock Size and Source on Ovarian Maturation and Spawning of Penaeus monodon Fabricius from the Gulf of Thailand

The effects of broodstock source (pond-reared and wildcaught from shallow waters) and prawn size (large—weight >120 g, and small-weight < 110 g) on ovarian maturation and reproductive success of Penueus monodon were evaluated. Large females underwent stage IV ovarian maturation and spawned with greater success than did small prawns. Both pond-reared and wildcaught females of the same size exhibited comparable maturation and spawning success. Large females rematured and spawned more frequently than small ones. Total eggs produced by large prawns was significantly greater than for small prawns; nevertheless, prawn source and size had no significant influence on amount of eggs spawned per spawning event. Egg quality varied greatly in terms of percent fertilization, hatch rate, and metamorphosis to first protozwa stage.     

Effects of dietary calcium and phosphorus supplementation on the growth performance of juvenile spotted babylon <Emphasis Type="Italic">Babylonia areolata</Emphasis> culture in a recirculating culture system

A feeding experiment was conducted to determine the effects of dietary calcium and phosphorus, and the interaction between calcium and phosphorus, on the growth of juvenile spotted babylon, Babylonia areolata, cultured in a recirculating culture system. Nine isonitrogenous experimental diets supplemented with three levels of calcium (1, 4, and 7%) for each of three levels of phosphorus (1, 3, and 5%) were prepared using fish meal, squid meal, and shrimp meal as the main protein sources. Juveniles with an initial average body weight of 0.59 ± 0.09 g were fed to satiation once daily with one of the nine diets for 180 days. Absolute and specific growth rates were calculated for both shell length and whole wet body weight. Results showed that dietary calcium and phosphorus supplementation significantly affected the growth of juvenile spotted babylon (P < 0.05), but not survival and feed-conversion ratio. The specific growth rate in shell length (SGRL) ranged from 0.32 to 0.39% day⁻¹. No significant difference among phosphorus levels and no significant interaction between calcium and phosphorus in SGRL of the spotted babylon (P > 0.05) was found, but significant differences were observed among calcium levels, irrespective of phosphorus levels (P < 0.05). For 1 and 7% supplemental calcium, the spotted babylon had significantly higher SGRL than those fed diets supplemented with 4% calcium. However, the specific growth rate in body weight (SGRW) ranged from 0.91 to 1.19% day⁻¹ with no significant difference among calcium and phosphorus levels and no significant interaction between calcium and phosphorus (P > 0.05). Survival and feed-conversion ratio were not significantly affected by dietary calcium and phosphorus levels with ranges from 91.00 to 95.00% and 2.43 to 2.76, respectively. At the end of the experiment, shell abnormality of B. areolata was found for all feeding trials.     

Differences in fatty acid composition of egg capsules from broodstock spotted babylon, <Emphasis Type="Italic">Babylonia areolata</Emphasis>, fed a local trash fish and formulated diet under hatchery conditions

This study is the first attempt to condition broodstock Babylonia areolata using formulated diets under hatchery conditions. Samples of spotted babylon egg capsules from broodstock fed either a formulated diet or a local trash fish, carangid fish (Seleroides leptolepis) for 120 days were analyzed for proximate composition and fatty acid composition. The formulated diet contained significantly higher levels of arachidonic acid (20:4n − 6; ARA), eicosapentaenoic acid (20:5n − 3; EPA) and docosahexaenoic acid (22:6n − 3; DHA) than those of the local trash fish. The formulated diet also had significantly higher ratios of DHA/EPA and (n − 3)/(n − 6) PUFA than those of local trash fish but not for the ARA/EPA ratio. The compositions of egg capsules produced from broodstock fed formulated diet contained significantly more ARA, EPA and DHA compared to broodstock fed the local trash fish. The ARA/EPA and DHA/EPA ratios in egg capsules were significantly higher in the trash fish—fed group compared to those fed the formulated diet. However, (n − 3)/(n − 6) PUFA ratios in egg capsules produced from broodstock fed the formulated diet did not differ significantly compared to those from broodstock fed the local trash fish. The relatively low DHA/EPA, ARA/EPA and (n − 3)/(n − 6) ratios in the egg capsules produced from the formulated diet—fed broodstock B. areolata suggested that this diet is inferior, when compared to the traditional food of trash fish.     

Effect of dietary supplementation of brewers yeast and nucleotide singularly on growth,survival and vibriosis resistance on juveniles of the gastropod spotted babylon (<Emphasis Type="Italic">Babylonia areolata</Emphasis>)

This study was undertaken to evaluate the use of brewers yeast and nucleotides as a growth promoter and to provide vibriosis resistance for the juveniles of gastropod spotted babylon (Babylonia areolata). Juvenile spotted babylon (0.3 g initial weight) were randomly distributed at a density of 50 snails in 45-L aquaria and fed a basic diet (40% crude protein) containing two incremental levels of 1 and 2% brewers yeast and nucleotides singularly for 4 months. After the feeding trial, snails from each treatment were challenged by pathogenic bacteria Vibrio alginolyticus given by intramuscular injection and kept under observation for 5 days to record clinical signs and daily mortality rates. Results indicated that the snails fed with diets supplemented with brewers yeast or nucleotides exhibited significantly greater growth than those fed the basic diet (P < 0.05) and significantly better food conversion ratios compared to snails fed the basic diet (P < 0.05). These results indicated that dietary supplementation of brewers yeast or nucleotides, at least at the tested dosages, enhanced spotted babylon growth. Supplementing the diet with 1% brewers yeast promises to provide appropriate resistance to V. alginolyticus.     

Freeze‐dried forms of mosquito larvae for feeding of Siamese fighting fish (Betta splendens Regan, 1910)

Live diets are preferably used for rearing Siamese fighting fish (Betta splendens Regan, 1910) since they provide superior growth over the dry feed. In the current study, three different preparations of mosquito larvae were prepared (frozen at ?20°C, F–20; freeze‐dried and kept at 4°C, FD4; freeze‐dried and kept at ambient temperature, FDAT) and were individually fed to 1‐month‐old red male fighting fish (1.18 ± 0.01 g initial body weight) over 6 weeks duration. At the end of experiment, there were no significant differences in growth performance and feed utilization across three dietary treatments (p ? 0.05). Specific activity of lipase was significantly lower in fish fed FD4 and FDAT than with the F–20 diet, while no differences in other enzymes were observed. The fish fed with FDAT diet significantly increased in viscerosomatic index relative to F–20 and FD4 treatments. Significant improvements in skin redness and flesh quality (RNA and RNA/protein ratio) were observed in the fish fed with FDAT diet relative to the other treatments. This preferred FDAT treatment also maintained the carcass composition. Analysis of digestive enzymes in FDAT mosquito larvae demonstrated the presence of protein‐, carbohydrate‐, and lipid‐digesting enzymes after 1 month of storage. The findings from our experiments indicate that the freeze‐dried form (FDAT) of mosquito larvae is suitable for rearing Siamese fighting fish. However, effective preparation protocol and appropriate storage times should be further studied.