In this study, the PHB-accumulating Bacillus sp. JL47 strain (capable of accumulating 55% PHB on cell dry weight) was investigated for its effects on the immune response of giant tiger shrimp (Penaeus monodon) postlarvae (PL) before and after the Vibrio campbellii challenge. Briefly, shrimp PL were cultured and fed with Artemia nauplii enriched with Bacillus sp. JL47. Shrimp receiving the Artemia nauplii without JL47 enrichment were used as control. After 15 days of feeding, the shrimp were challenged with pathogenic V. campbellii LMG 21363 at 10⁶ cells mL^-1 by immersion. Relative expression of the immune related genes encoding for prophenoloxidase (proPO), transglutaminase (TGase) and heat shock protein 70 (Hsp70) in the shrimp were measured before (0 h) and after (3, 6, 9, 12, 24 h) the Vibrio challenge by quantitative real-time PCR using β-actin as the reference gene. The expressions of TGase and proPO were significantly up-regulated (p < 0.05) within 9 h and 12 h, respectively after challenge in shrimp receiving the Bacillus sp. JL47 as compared to the challenged and non-challenged controls. Hsp70 expression was significantly increased (p < 0.05) at 3 h post-challenge in all challenged shrimp. Interestingly, proPO and TGase genes were significantly up-regulated (p < 0.05) in Bacillus sp. JL47 treated shrimp even before the Vibrio challenge was applied. No up-regulation in the Hsp70 gene, however, was observed under these conditions. The data suggest that the protective effect of the PHB-accumulating Bacillus sp. JL47 in shrimp was due to its capacity to stimulate the innate immune related genes of the shrimp, specifically the proPO and TGase genes. The application of probiotic Bacillus species, capable of accumulating a significant amount of PHB, is suggested as potential immunostimulatory strategy for aquaculture.

The addition of artificial shelters of various materials has been used in Macrobrachium rosenbergii tanks and ponds as a means of increasing productivity. The present study investigated the shelter colour preference of M. rosenbergii postlarvae (age 15–18 days after metamorphosis) in the laboratory. Shelter occupancy tests were performed on four groups of 200 postlarvae in four 57-l aquaria, into which were placed shelters made of rigid coloured netting sewn into four-layered cubes open on two sides. The shelters were presented in six colour pairs: black vs dark green, black vs light green, black vs blue, dark green vs light green, dark green vs blue, and light green vs blue. Colour preference was tested six times for each colour pair. The data obtained were analyzed using Thurstone’s law of comparative judgment. The mean z-score was significantly highest (i.e., highest preference) for the black shelter. Vision was involved in the detection of and approach to the shelter. When released in aquaria, intact-eyed individual postlarvae directly approached the shelter straight away, whereas blind postlarvae (paint over the eyes) swam around randomly and occupied the shelter only after accidental contact much later. Postlarvae in open water out of the shelter exhibited frequent aggressive contact, while those in the shelter were quiescent. The use of black shelters in rearing tanks is thus recommended for reducing stress and aggression among M. rosenbergii postlarvae in the hatchery.

Luminous vibriosis is a devastating infection of penaeid shrimp larvae and juveniles causing heavy mortalities. To counter the bacterial pathogen, Vibrio harveyi, shrimp farmers in the Philippines modified their growout culture method of the black tiger shrimp, Penaeus monodon, juveniles and developed the greenwater culture technique. This culture method involves the use of pond water of all-male, saline-tolerant Oreochromis hornorum as rearing water for the culture of shrimp juveniles in ponds. Such a modified culture of P. monodon juveniles was found effective in preventing the onset of luminous vibriosis. Basic studies revealed that antiluminous Vibrio factors are inherent in the bacterial, fungal, and microalgal flora of the tilapia water, dermal mucus, and gut that singly or collectively inhibit the growth of V. harveyi, in vitro. The skin mucus studies of other brackishwater fish species showed that the siganids, Siganus guttatus, and red hybrid tilapia (Oreochromis niloticus × Oreochromis mossambicus), as well as sea bass, Lates calcarifer, are promising alternative fish species for this novel shrimp culture method. A review of pond-simulated studies in tanks and ponds, similarly, confirmed these findings and the impact of the greenwater culture technique on water quality, including its economic benefits to the farmer. The greenwater culture of shrimp can sustain the successful production of shrimp juveniles by inhibition of the luminous Vibrio. This culture method is also currently used in the growout culture of the white shrimp, Litopenaeus vannamei.

Changes in innate immunity parameters and epinecidin mRNA transcript levels were examined to characterize the non-specific immune response of E. coioides to pathogenic V. harveyi JML1 isolated from affected cage-cultured fish. After fish had been injected with bacteria at a dose causing 30% mortality, blood and tissue samples were collected at 0, 6, 12, 24, 48, 72, 96, 120, and 240 h post-infection (hpi) for assessment of indices such as the oxidative burst (OB) and phagocytic index (PI) of head kidney cells, and lysozyme activity (LYS) and total immunoglobulin (Total Ig) levels of the plasma. The epinecidin mRNA transcript levels (EGE) from skin, gills, liver, kidney, and spleen tissues were also determined by gelbased RT-PCR. Lastly, daily mortality (DM), liver total bacterial load (TBC), and presumptive Vibrio count (TVC) were monitored up to 240 hpi. The results revealed that bacteria proliferated rapidly in fish tissue, reaching peak densities at 24 hpi for both TBC and TVC but was on a downward trend thereafter. The pattern in fish mortality closely correlated with TBC and TVC. Total Ig, OB, and PI in E. coioides were suppressed in the early part of infection when V. harveyi load was high but recovered and later increased as bacterial density declined. LYS and EGE were consistently high and their activities were not hampered by bacterial infection. The study demonstrated that V. harveyi JML1 interacts with E. coioides by transiently inhibiting some immune parameters resulting in mortalities. However, consistently high LYS, upregulated EGE, and resurgent PI, OB and Total Ig conferred resistance and subsequent recovery in the fish. The study provides new insights on the interaction between E. coioides and V. harveyi JML1 that can aid in formulating health management strategies for groupers. Further studies on prophylactic interventions to enhance the innate immune response in grouper during infection with V. harveyi JML1 are suggested.

A 90-day feeding trial was conducted on milkfish, Chanos chanos with an initial mean body weight of 3.07 ± 0.17 g (mean ± standard error of mean). Six treatment diets were formulated to contain 0 g/kg (Diet 1), 150 g/kg (Diet 2), 25 g/kg (Diet 3), 300 g/kg (Diet 4), 350 g/kg (Diet 5) and 450 g/kg (Diet 6) distiller's dried grain with soluble (DDGS). All the dietary treatments were isonitrogenous (350 g/kg crude protein) and isolipidic (6% crude lipid). Result of the feeding trial indicated that growth rates, feed intake and feed efficiency were not significantly (p > .05) affected by inclusion levels of DDGS by up to 450 g/kg in the feed. Proximate body composition (crude protein, crude lipid, ash, fibre) in fish fed the dietary treatments were not significantly (p > .05) affected as well. The DDGS when used as a milkfish ingredient has a protein digestibility of 910 g/kg, fat disgetsibility of 850 g/kg, carbohydrate digestibility of 750 g/kg and a dry matter digestibility of 520 g/kg Results from the intestinal morphology displayed no apparent pathological changes in the digestive tract of fish fed all dietary treatments. These results indicate that DDGS can be efficiently utilized by milkfish by up to 450 g/kg without negatively affecting performance parameters and intestinal morphology.

Starvation and exposure to formalin were investigated as possible stress tests for evaluating the quality of mud crab, Scylla serrata, larvae. For the starvation stress test, newly hatched zoeae stocked in 150-ml containers were either starved or fed rotifers. Similarly, newly hatched zoeae were stocked in containers with seawater of 0 (control), 20, 30 and 40 mg/L formalin for the formalin stress test. The zoeae from the same batches were used for seed production to monitor their performance and validate the results of stress tests. Starvation was found to be unsuitable for larval quality evaluation. However, the impact of initial food deprivation on the newly hatched larvae indicates that feeding immediately after hatching is necessary for mud crab larvae. Exposure of larvae to 40 mg/L formalin for 3 hr appeared to be a reliable and practical method for larval quality assessment as the survival of larvae in the mass production tanks validated the classification of good and poor quality batches in the stress tests. On this basis, a hatchery operator can decide which batch should be cultured further. Finally, there appears to be a link between the quality of larvae and the performance at the megalopa and early juvenile crabs.

The abdominal segment deformity disease (ASDD) is a new shrimp disease reported only in cultured Penaeus vannamei in Thailand. Shrimp with ASDD have deformed abdominal segment, jagged gut line and bumpy surfaces. Similar signs were observed in cultured P. indicus in the Philippines. However, aside from the signs described for ASDD, some P. indicus showing abdominal segment deformity syndrome (ASDS) had more severe deformities up to the extent that the number of body segments was reduced due to fusion. Shrimp with fused body segment deformity (FBSD) had four instead of five pairs of legs. To account the prevalence of the deformities in P. indicus, shrimp were classified into grossly normal shrimp (NS), shrimp with abdominal segment deformity syndrome (ASDS) and shrimp with fused segments (FBSD). Out of the shrimp sampled, 83.4 ± 5.4% was NS, 10.9 ± 6.2% was ASDS and 5.7 ± 3.0% was FBSD. Morphometric characteristics of the shrimp were measured. There was no significant difference in body weight (BW) among male and female NS, ASDS and FBSD. In both sexes, total length (TL) of FBSD was significantly shorter compared to NS and ASDS. Shrimp samples were also screened to be negative for known infectious viral diseases including white spot syndrome virus (WSSV), infectious hypodermal and haematopoietic necrosis virus (IHHNV), infectious myonecrosis virus (IMNV), P. vannamei nodavirus (PvNV), Macrobrachium rosenbergii nodavirus (MrNV) and Taura syndrome virus (TSV). Occurrence of ASDS and FBSD in post-larvae (PL) produced from captive and wild spawners were also determined. Based on a tank experiment, no significant difference was detected between the percentages of ASDS in PL produced from wild or captive spawners but FBSD was only noted in PL produced from the latter. Deformities generally did not affect the size of P. indicus except for the reduced length of shrimp with FBSD which when coupled with missing pleopods could lead to major economic loss for shrimp farmers if not addressed properly.

Development of economically viable techniques for growing hatchery-reared juvenile crabs to suitable sizes will address the problem on the source of seed stocks for soft-shell crab farming. This paper reports the production of hatchery-reared mud crab Scylla serrata from juveniles in the nursery to 73-106 g body weight (BW) crabs in the grow-out pond for the individual system soft-shell crab farming. Likewise, the performance of hatchery-reared S. serrata, and wild S. tranquebarica and S. olivacea juveniles was determined in the soft-shell crab production set-up.

The BW increased from 1.8-1.9 g to 78-113.7 g when stocked at 0.5 ind m^-2 and from 1.6-2.3 g to 73-106.7 g at 1.0 ind m^-2 after 75 days. Growth rates at both stocking densities were comparable. However, survival was significantly higher (P<0.05) in lower (63.6~c1.01%) than in higher (35.6~c3.34%) stocking density. Male S. serrata (46.0 ~c 1.75%) had significantly higher BW increase than females (39.4 ~c 2.05%). Crabs stocked at sizes of 51-60 g showed significantly greater percent increase in BW (43.26~c 0.98%) compared with those at 61-70 g (40.98~c1.33%), 71-80 g (38.55~c 1.04%), 81-90 g (36.34 ~c 1.27%) and 91-100 g (38.52 ~c 1.67%). Among the three species, hatchery-reared S. serrata (42.14 ~c 1.34%) had significantly higher mean percent BW increase compared with S. olivacea (38.23 ~c 0.49%) and S. tranquebarica (36.16 ~c 0.78%). S. serrata had significantly shorter mean culture period (24.11 ~c 0.95 days) than S. tranquebarica (28.48 ~c 0.54 days) and S. olivacea (28.75 ~c 0.34 days).

Studies were conducted towards the pilot scale production of stable and nutritionally adequate pellets suitable for mud crab Scylla serrata to help in efforts to increase mud crab production in the Philippines. Preliminary studies showed that mud crabs preferred pellets which were spherical or cylindrical and contained marine based attractants. Two practical diets were formulated and prepared in the Pilot Feed Mill of SEAFDEC/AQD. The extruded diets, along with a commercial mud crab grow-out diet, were subjected to proximate and Ca/P analyses as well as cholesterol analysis and water stability tests. The diets were fed to crabs in a controlled laboratory experiment using fiberglass tanks with flow-through, aerated seawater, and monitored from initial molt (M0) up to the second molt (M2). Tests revealed that the crabs fed Diet 1 (CP 48.28 %, CF 7.74 %, Ca 5.23%, P 12.93 %, cholesterol 0.173 %) showed the highest growth and better survival than those fed Diet 2 or commercial diet. Crabs fed Diet 1 took a shorter time to attain two molt cycles from M0. Survival was 70% after M2. Diet 1 was water stable within two hours. Abnormalities were observed including absent swimming legs or chelae, exposed gills, sores on exoskeleton, incomplete molting, or soft exoskeleton, but none that could be directly attributed to a specific treatment. Incomplete molting and slow hardening of the exoskeleton in crabs that accounted for the mortalities could be due to the low cholesterol levels in the diets especially in the commercial diet which had the lowest cholesterol level. Cholesterol supplementation of the artificial diet would be needed to improve molting success and survival of mud crabs in grow-out culture. The results of these experiments will be used to formulate a cost-effective grow-out diet for mud crab (supported by DOST-PCAARRD Grant in Aid).

This study aims to improve the handling, storage and transport conditions of mud crabs. The usual practice of the crab growers in the Philippines is to bring the market size crabs to middlemen, brokers or operators of small trading centers immediately after harvest; then the crabs are brought to municipal traders or operators of bigger buying station. The crabs are sent to the exporters in Manila or Cebu. The crabs are exported mostly to Singapore, Taiwan, Hongkong and mainland China. The crabs are classified based on the species, body weight, sex, gonad maturity and intactness of limbs. Crabs are rejected for export when they are soft-shelled, very lean, or have incomplete limbs and abnormalities. Crabs with emaciated muscle (‘hagas’), ammoniacal odor (strong urine-like smell) and in an undesirable state are also rejected.

This paper presents the initial results of the project on the Improvement in the handling, storage and transport of mud crabs under Sub-program C entitled Improvement of Feeds and Stock Management Practices in Mud Crab Grow-out Culture under the National Mud Crab Science and Technology Program. It also presents issues and concerns on the present practices, and presents possible recommendations.