Rapid detection of white spot syndrome virus (WSSV) of Penaeus monodon by latex agglutination test using monoclonal antibodies

Latex beads were sensitized with monoclonal antibodies (MAb) rose against VP28 of WSSV. The optimum concentration of MAb required to sensitize the latex beads was 125 µg/ml. The sensitized latex beads were used to detect WSSV from PCR-positive stomach tissue homogenates obtained from infected shrimp. Stomach tissue homogenates from WSSV-infected shrimp agglutinated the sensitized latex beads within 10 minutes, while uninfected samples did not produce any agglutination, although non-specific agglutinations were observed in some samples. The analytical sensitivity, analytical specificity, diagnostic sensitivity and diagnostic specificity of the (LAT) agglutination test were assessed. The analytical sensitivity of the test was 40 ng of purified WSSV (2 µg/ml). The sensitized latex beads did not agglutinate with normal shrimp tissue or MBV-infected tissue homogenate. The test has a diagnostic sensitivity of 70 and 45%, respectively, compared to single-step and nested PCR. The diagnostic specificity of the test was 82%. This test is a simple and rapid on-farm test which can be used to corroborate clinical signs for the detection of WSSV in grow-out ponds.

Feasibility of bi-culture of mud crab (Scylla serrata) and shrimp (Penaeus monodon)

A study on the feasibility of bi-culture of mud crab (Scylla serrata) and shrimp (Penaeus monodon) in brackishwater earthen ponds (0.1 ha each) was carried out for a period of five months (March-August). Nursed shrimp juvenile (ABL:· 3.36±0.23 em and ABW: 0.26±0.04 g) and crab juvenile (ACL: 2.61±0.22 cm, ACW: 4.63±0.11 cm and ABW: 43±2.64 g) were stocked following the experimental design of shrimp 2/m2 (Treatment-1), shrimp 2/m2 and mud crab l/m2 (Treatment-2) and shrimp 2/m2 and mud crab 0.5/m2 (Treatment-3). Crabs were fed with chopped trash tilapia @ 10~5%, while shrimp were fed with Saudi-Bangla shrimp feed @ 3~5% of biomass twice daily. Significantly (p<0.05) higher specific growth rate (SGR) of shrimp and mud crab was 1.86% (g/day) in T2 and 0.83% (g/day) in T3, respectively. The survival of shrimp and mud crab also varied significantly (p<0.05) with a higher mean value of74.63% in Tl and 51.04% in T3, respectively. The production of shrimp (424.09 kg/ha) was significantly (p<0.05) higher in Tl and that of mud crab (568.80 kg/ha) in T2. Significantly (p<0.05) highest total production of 871.29 kg!ha was in T2 followed by 708.52 kg/ha in T3 and 424.09 kg/ha in Tl. The results indicate that mud crab can be cultured at a stocking rate of 1/m2 together with shrimp at 2/m2 •

Genetic analysis of east and west coast populations of Penaeus monodon from India based on random amplified polymorphic DNA

The east and west coast populations of wild Penaeus monodon in India were genetically characterized by RAPD analysis using six highly polymorphic primers reported earlier. The average genetic similarities within populations, based on profiles generated by all the six primers, were 0.828 and 0.851 for the east and west coast populations, respectively, values with individual primers ranging from 0.744 to 0.889. The average genetic similarity between populations across all the primers was 0.774. The number of bands found to be polymorphic were 38 (51.35%) and 37 (50.68%) in the east and west coast populations, respectively. Primer 5 yielded the highest level of polymorphism (63.63%) in the east coast population whereas primer 3 yielded the lowest level of polymorphism (36.36%) in the west coast population. The study reveals the existence of genetic variation in P. monodon stocks providing scope for genetic improvement through selective breeding. It also provides baseline data for future work on population structure analysis of P. monodon.

Utilization of trash fish for human consumption. 3: Studies on the development of fish pickle from Nemipterus japonicus

A method for the preparation of fish pickles from a lean variety of fish namely pink perch (Nemipterus japonicus) is described. Dipping the fish in 10% sodium chloride solution containing 6% acetic acid before pickling, was found desirable for retaining the meaty texture of the product. The product has no fish smell or flavour and has a shelf life of more than six months at ambient temperatures and scored very well in organoleptic tests.

Preliminary study on the rearing of Chilka prawn ('Kantala') Penaeus indicus H. Milne-Edwards in the brackish water ponds at Keshpur (Orissa)

Results of experimental rearing of Penaeus indicus without supplemental feeding in unfertilized brackish water ponds in fore-shore of Chilka lake are presented in this paper. Higher rate of survival was recorded where advanced juveniles were stocked and lower rate of survival was recorded where early juveniles were stocked. The average growth rate recorded from the rearing experiments was higher than the average growth rate of Penaeus indicus recorded from the adjoining Chilka Lake which indicates the promising future of prawn farming in and around the lake. Analysis of gut contents revealed that Penaeus indicus could efficiently utilise algal and higher plant matter present in the ponds. No noticeable differences in daily growth rates were noticed between early and advanced juveniles in rearing experiments.

Late Pleistocene divergence and subsequent population expansion of two closely related fish species, Japanese anchovy (Engraulis japonicus) and Australian anchovy (Engraulis australis)

Climatic oscillations during the Pleistocene ice ages produced great changes in species' geographical distribution and abundance, which could be expected to have genetic consequences. Living in the temperate upwelling zones of the northwestern Pacific, Ja

Differential population structuring and demographic history of two closely related fish species, Japanese sea bass (Lateolabrax japonicus) and spotted sea bass (Lateolabrax maculatus) in Northwestern Pacific

The Quaternary cold periods in the Northwestern Pacific are thought to have heavily influenced the amount and distribution of intraspecific genetic variation in marine fishes. To estimate the demographic history and genetic structure of Lateolabrax macula

Succession of bacterial genera during iced storage of three species of tropical prawns, Penaeus indicus, Matapenaeus dobsoni and M. affinis

The native bacterial flora of ocean fresh tropical prawns, Penaeus indicus, Metapenaeus dobsoni and M. affinis was more or less similar, mainly consisting of Pseudomonas, Acinetobacter, Moraxella and Arthrobacter. A definite succession of bacterial genera during iced storage was observed in these prawns. As the day of ice storage increased, the proportion of Acinetobacter and Moraxella also increased considerably and constituted 70-78% of the flora at the time of spoilage. Spoilage by Pseudomonas was very not significant in prawns under iced storage.

Identification and culture of common diatoms as possible feed for Penaeus monodon

Diatoms were collected from Buyuan Bay, and from the hatchery tanks at Tigbauan, to determine the commonly occurring species, the feasibility of culturing these species, and the potential of these selected species as food for larval P. monodon. The commonly occurring diatoms were identified as Chaetoceros calcitrans, Navicula grimmei, Nitzchia seriata, Nitzchia closterium and Amphiprora sp. These diatoms were isolated and unialgal cultures prepared. Protein content analysis using the micro-Kjildahl method gave the following result: C. calcitrans, 11 . 78%; Nitzchia seriata, 25%; Nitzchia closterium, 30 . 5%; Navicula grimmei, 9 . 06% and Amphiprora sp. 8 . 96%. Feeding experiments were conducted to determine acceptability of the different diatom species and percentage survival of larval stages Z SUB-1 -M SUB-2 . Larvae were placed in 4-l capacity plastic containers with a stocking density of 10/l. The results of several feeding trials using the different mass-produced diatoms are summarized. From the data gathered, C. calcitrans appears to be the most promising candidate as feed for zoea and mysis stages of P. monodon. The average percentage survival of C. calcitrans was 63 . 76% for the 3 trials, and as high as 82 . 22% in the third trial. Comparatively high percentage survival of larvae was also recorded when Nitzchia seriata (48 . 17%) and Nitzchia closterium (67 . 6%) were given as feed, while both Amphiprora sp. and Navicula grimmei gave 0% survival. The poor results with Amphiprora sp. and Navicula grimmei may be due to their low protein content (8 . 96% and 9 . 06%, respectively) and the inability of the larvae to ingest them. Navicula and Amphiprora were observed to cling to the appendages of the larvae and to settle down in the medium making them unavailable to the larvae. Low survival was also noted when frozen C. calcitrans was used (14 . 25%). This may be due partly to the effect of the floculating agent (ALSO SUB-4 . 25 g/l) used in concentrating the diatoms. When protein contents of C. calcitrans, N. seriata and N. closterium are compared, the 2 Nitzchia species have relatively higher protein contents than C. calcitrans and, therefore, could be the more desirable feed candidates. However, few feeding trials were made using Nitzchia so that additional investigations will have to be done on this aspect.

A suctorean parasite of Penaeus monodon larvae

A pathogenic suctorean, identified as Ephelota gemmipara was observed in P. monodon larvae spawned and reared in tanks. Commonly found to inhabit hydroid colonies, E. gemmipara has a stalked body with two types of tentacles, the sucking and piercing types, and was observed to reproduce by multiple exogenous budding.

Effect of furanace on the development of larval stages of Penaeus monodon Fabricius

Zoea 2(Z SUB-2 ) Mysis 1 (M SUB-1 ) and Postlarva 1 (P SUB-1 ) of P. monodon artificially spawned in closed-system concrete hatchery tanks were bioassayed for their tolerance to the antibiotic furanace. The setup consisted of four 20-liter capacity plastic basins previously conditioned for 15 days with freshwater in full sunlight. During the experiment, each basin was filled with 5 liters of seawater to which was added filtered Chaetoceros and Brachionus to give densities of 5 . 0-7 . 5 x 10 SUP-4 cells/ml and 10-20 individuals/ml, respectively. The following are the properties of the water used throughout the experiments: salinity, 26-32%; pH, 7 . 3-8 . 4; temperature, 25-30 degree C; dissolved oxygen, 4 . 5-8 . 4 ppm; nitrite, 0 . 36-0 . 99 ppm; and ammonia, 0 . 10-0 . 30 ppm. To each basin were added 50 healthy larvae of specific stages of P. monodon. After an initial acclimation of one hour in the medium, preweighed amounts of the antibiotic were added and thoroughly dissolved. The concentrations tested were 1 . 0, 2 . 0 and 3 . 0 ppm. One basin always served as control. After 24 hours of exposure, the surviving population in each basin was counted. The survivors were then examined thoroughly under the microscope for unusual behavior and morphological defects brought about by the exposure. To minimize wide variations in the medium as a result of feeding and other manipulations, the systems were all prepared at 9:00 a.m. each time, and the feeds on two instances, one at 5:00 p.m. and another at 5:00 a.m. Fifteen trials conducted with Z SUB-2 showed survival ranges of 68% to 98% with a mean of 77 . 6% in the controls; 32% to 94% with a mean of 65 . 7% at 1 ppm, and 0% to 56% with a mean of 36 . 5% at 2 ppm. There were no survivors at 3 ppm. Interpolation from the survival-dose curve gave a 24-hr LC SUB-50 of approximately 1 . 6 ppm.

Tolerance of Penaeus monodon larvae to cupric sulfate added in bath

Copper is used to deter the growth of bacterial, fungal and protozoan disease organism in fishes. Zoeae (Z SUB-1 ), myses (M SUB-1 ) and postlarvae (P SUB-1 ) were exposed to copper sulfate at concentrations of 0 . 025, 0 . 05, 0 . 75, 0 . 1 and 0 . 2 ppm from 24 to 96 hours. The number of surviving larvae were counted at the end of each 24-hour period and the percentage of survival is determined for each dose level. The LC SUB-50 for each of the larval stages was interpolated from the data whenever possible. Three trials with 2 replicates per trial were conducted. The physico-chemical characteristics of the bath taken before and at the end of the experimental period show insignificant differences between initial and final values in each trial. Results indicate that mortality rates of all larval stages increased with exposure time and that mortality rates of the experimental group is higher than the control. Interpolation of the LC SUB-50 is possible only for the 48-h and 72-h exposure times for both zoeae and myses and for the 48-h exposure time for the postlarvae. This is due to the high survival percentage of the 24-h group and the low survival percentage (below 50%) of the larvae exposed for 96 hours. The 48-hour LC SUB-50 for Z SUB-1 , M SUB-1 and P SUB-1 are 0 . 225, 0 . 350 and 0 . 125 ppm respectively. Postlarvae seem to be more sensitive than either of the 2 larval stages having a lower 48-h LC SUB-50 and a low survival rate after 72 hours. The larvae were observed to lose their balance and were lethargic, producing few swimming movements so that they were mostly confined to the bottom of the aquaria. Moribund larvae observed under the microscope had a faster but weak heartbeat compared to healthy larvae. Slight or complete loss of feeding ability indicated by empty guts and delayed molting of Z SUB-1 to Z SUB-2 were also noted.

The effect of various protein levels on the growth and survival rates of Penaeus monodon Fabricius

Postlarvae of tiger prawns, P. monodon , were fed with various protein levels of 0; 30; 40 and 50%, fish meal and rice bran were combined and pelletized for 16 wks. Prawns used had an average mean length of 1.2 mm and an average mean weight of 5.5 mg. Nitrite, ammonia, dissolved oxygen content, temperature and pH were monitored. After one month of feeding, growth increments of postlarvae fed with 30% protein diet were significantly different from those given 0; 40 and 50%. Survival rate of the group fed 40% protein was higher (81.7%) but was not significantly different from those fed 30% (68.0%). The highest feed efficiency was obtained from the 30% protein pellets (4.6). At the end of 16 weeks, growth increment of 40% protein level was significantly different from the 30% and 50% protein levels, but the survival rate was not signficantly different from the 30% protein level. Feed conversion for the 40% protein diet was 2.8.

Notes on the induced maturation and spawning in four-month-old Penaeus monodon Fabricius by eyestalk ablation

The ablation technique consisted of making an incision across the eyeball to allow free flow of fluids while holding the prawn under water, squeezing the eyeball contents outwards, and pinching hard the eyestalk tissue. The cut area heals completely in about a week; no application of antibiotics is necessary. Spent spawners were tagged with thin brass rings (Rodriguez, 1976) around the unablated eyestalk for a separate experiment on rematuration. Two spawning yielding approximately 277,000 eggs were obtained three weeks after ablation, followed four days later by two more spawnings with 160,000 eggs; all four spawners weighed more than 100 g. With a hatching rate of 98% and 78% for the first and second batch, respectively, the spawnings produced viable nauplii. Water temperatures as low as 23 degree C due to a delayed cold spell in March depressed molting; weakened larvae had to be discharged at the mysis stage. Although ovarian development continued, no further spawnings were obtained due mainly to the onset of bacterial and fungal disease. Infection is initiated in injured portions of the exoskeleton, sometimes penetrating right through the muscles to the ovarian tissues. The non-flowthrough conditions and mussel meat feeding led to fouling of the culture water resulting in consecutive mortalities caused by disease. Female P.monodon held in maturation pens were ablated at the age of 15 months (Santiago, et al., 1976); they averaged only 16 g body weight after four months growth in ponds. In another experiment, pond-reared P.monodon females ranging from 50 to 80 g were ablated at approximately seven months (Aquacop, 1977). The present results show a minimum age of four months from postlarve that P.monodon is capable of ovarian development and spawning upon ablation. However, maturation is probably affected by size as well as age - the four-month old females weighed an average of 100 g in contrast to the smaller animals in the earlier experiments.