Culture and Management of Scylla Species (CAMS)

Work Package 1: Bacterial Disease Control

Objective: To improve the reliability and sustainability of crab hatchery systems through the use of probiotic bacteria as an
alternative to microbials in disease control

PHILIPPINES

For 2002 and 2003, monitoring activity of problems in large-scale hatchery rearing of crab larvae was
done to identify windows of opportunity for probiotic application. These led to identification of various
microbial fouling and disease-causing organisms in spawned eggs and hatchery-reared larvae.

Microbial populations in eggs and larvae

Disease problems in mud crab larval rearing include severe fouling of eggs and larvae with filamentous
algae, protozoan infestation, and sporadic fungal infection in the egg masses. Mortality of larvae in the
hatchery is due to systemic bacterial infection and occasionally, fungal infection. Based on several runs,
the main sources of bacterial pathogens in the hatchery are the following: developing eggs, untreated
water supply, and natural food, mainly Brachionus and Artemia. Although the water supply can be
effectively rid of bacterial pathogens by chlorination (10-15 ppm active chlorine), such technique does
not remove pathogenic bacteria from other sources.

Severe fouling of eggs with filamentous alga Protozoan infestation in eggs

Fungi that were isolated from mud crab eggs and larvae are of two general types, vesicle-formers
(Lagenidium-like) and non-vesicle-formers (Sirolpidium-like). For both fungal types, spore formation
and release were inhibited by 10 ppm formalin bath exposure for 24-40 h, although the fungi remained
viable and grew when replanted on peptone yeast extract agar. Fungal hyphae exposed to 50 and 100
ppm formalin for the same period did not produce nor release spores and were not visible after exposure.

Crab egg with fungal infection Zoe infected with fungi

White spot syndrome virus (WSSV) in megalopae and pond-reared juveniles

The WSSV carrier status of hatchery-reared crab megalopae was determined using one-step and nested
PCR tests using the WSSV-specific primer pairs designed by Kimura et al. (1996). Two batches of
hatchery-reared megalopae were negative for WSSV after one-step and nested PCR tests. However,
moribund samples of pond-reared crabs were positive after nested PCR test. Pooled samples
representing hatchery and wild-sourced crabs obtained from ponds during harvest were negative for
WSSV by one-step and nested PCR.
 Work Package 2: Broodstock and Larval Quality

Objective: To improve larval quality and survival through manipulation of broodstock and larval nutrition

PHILIPPINES

Histological and biochemical analyses

Ovarian maturation in wild-sourced and pond-grown Scylla serrata was studied in terms of gross
morphology, histological appearance, and nutrient profiles. Ovarian maturation was classified into six
stages (immature to spent). During maturation, protein, lipids, minerals, and ascorbic acid were
accumulated in the ovaries, which suggests that these nutrients are needed in reproduction. In wild-
sourced crabs, the essential amino acids arginine, leucine, methionine, and valine increased between the
immature and mature ovaries; triglycerides and phosphotidylcholine were accumulated; and the
concentrations of highly unsaturated fatty acids (docosahexaenoic acid, eicosapentaenoic acid, and
arachidonic acid) were high. Lipids decreased as eggs hatch to zoea indicating the importance of lipids
during embryonic development. Information on nutrient profiles will serve as basis for the improvement
of crab broodstock and larval diets.

Reproductive performance of S. serrata fed graded levels of dietary lipids with or without natural
food supplement

Adult pond-sourced S. serrata (625 + 53.3 g

body weight) were fed artificial diets with
squid oil and soybean lecithin (2:1 ratio) at
three levels (6, 9 and 12%) with or without
supplementation of natural foods (mussel,
squid or fish). Broodstock fed natural food
served as the control. After three months,
broodstock fed natural food alone and diets
with 6, 9 and 12% lipid with natural food
supplementation had better performance than S
those fed artificial diet alone. . serrata broodstock

Alternative live and artificial feeds for Scylla spp. larvae

Rotifers are routinely used as feed for crab larvae and as an alternative; the efficacy of oyster
(Crassostrea iridalei) trocophores was evaluated. Trocophores were acceptable as food but survival and
growth were better in larvae fed rotifers alone or trocophores in combination with Artemia.

Growth and survival of mud crab larvae fed artificial diets of various physical preparations (crumbled,
granulated, drum-dried and window refractance dried) were evaluated. All stages of zoea ingested the
artificial diets within 30 min of initial feeding with no difference to those given live food as control
(rotifers from zoea 1 to 5 + Artemia from zoea 3 to megalopa). The survival was higher in larvae fed
crumbled and granulated diets in two experiments using S. serrata and S. tranquebarica.

Functional and physiological maturity of Scylla spp.

Hatchery-reared S. serrata, S. tranquebarica and S. olivacea juveniles were grown to maturity in ponds,
meristic measurements were made, and the gonads and external sexual characters were monitored. The
abdomen width, carapace length, and propodus height, thickness and length were each plotted against
the internal carapace width (ICW). There is an increase in the abdominal width per unit increase in ICW
of females in the three species. The propodus height and thickness are greater in males than females per
unit increase in ICW. Mating readiness of S. olivacea occurs in smaller body sizes than those in S.
tranquebarica and S. serrata.

B. AQUASILVICULTURE Objective: To refine systems for integrated aquasilviculture of mud crabs within
mangroves, specifically by reducing fish biomass requirement through low cost incomplete feeds and utilization of natural
productivity
 PHILIPPINES

Feeding treatments: Wild S. olivacea

Grow-out mangrove pen systems suitable for coastal communities have been developed by AQD over
the years. Dependence on "trash fish" or fish biomass, used as food by low income groups, remains a
problem in farming mud crabs. To reduce if not completely eliminate such dependence, a low cost,
incomplete pellet was formulated. Wild S. olivacea (30-70 g BW) were stocked in pens with feeding
treatments as follows: A) no feeding (natural productivity), B) no feeding for 1 month followed by
supplementary feeding with cow/carabao hide, C) fish biomass, and D) incomplete pellets. Growth rates
peaked after three months then stabilized therafter, suggesting at this point selective harvest can be
started. Crabs fed fish and cowhide had significantly larger sizes after 3 months. They also showed the
highest total production, number of berried females and survival rates, although the latter was not
significantly different among treatments. Crabs with supplementary feeding survived on natural food in
the mangroves for the first month, indicating that artificial feeding of wild-sourced juveniles is not
necessary during this period. Pellet stability and crab retrieval need improvement.

Grow-out mangrove pen system for mudcrab

Varieties of S. olivacea

Two varieties of wild S. olivacea juveniles tentatively identified as 'morongsod' and 'sinaw' were stocked
separately in pens. 'Morongsod' are smaller, darker red and more aggressive than 'sinaw'. After four
months, body size and weight were generally higher for 'sinaw' than 'morongsod'. However, the number
of mature, berried and spawned females was higher for 'morongsod' compared to 'sinaw'. Genetic profile
of the two S. olivacea varieties needs to be confirmed.

Two varieties of Scylla olivacea juveniles-'sinaw' (left) and 'morongsod' (right)