Executive summary

TipTopp Aquaculture conducted a  trial to evaluate the efficacy of TIPTOPP’s products in post-larvae production of whiteleg shrimp  (P. vannamei, Boone, 1931) (1Holthuis, 1980). The objective of this trial is to demonstrate that the use of the products in combination with good hatchery protocols will significantly improve the production and quality of postlarvae (PL). 

The 22-day of larval rearing trial has been conducted at ShrimpVet hatchery. The trial consisted of three groups including control (T1), TIPTOPP fermentation (T2), and TIPTOPP direct use (T3). Post-larvae 10 (PL10) of all groups were taken to perform larvae health assessments and stress tolerance tests. 

The application of TIPTOPP direct use (T3) has a capable to enhance PLs production, reduce stress, and improve shrimp PLs quality. However, the innovative approach with fermenting the Tiptopp Pond also need to be adjusted and tested again to have better results in enhancing quality and cost effective in shrimp PLs production. 

1. Experimental products 

TIPTOPP provided ShrimpVet with test products (water bioremediation : “TIPTOPP POND” and in-feed probiotic: “TIPTOPP AQUAG”) for use in this trial. Guideline for the storage conditions and using instruction of the test products were also updated by TIPTOPP. 

• Fermented TIPTOPP POND Usage 

• • 2 liter fermented AquaFarm (1 gram per liter of mixture) per m3 tank volume before stocking Nauplii. 
  • 1 liter fermented AquaFarm (1 gram per liter of mixture) per m3 tank volume from Nauplii to Mysis 3. 
  • 2 liters fermented AquaFarm (1 gram per liter of mixture) per m3 tank volume from PL1 to PL10. 

• TIPTOPP POND Directly Usage 

• • 1 gram of TIPTOPP POND per m3 tank volume on daily basis. 

• TIPTOPP AQUAG Usage 
  • was mixed with artificial feed with dose rate 50 gram of TIPTOPP AQUAG per kg of artificial feed. 

2. Experimental design

The trial was set up as a completely randomized design (CRD) in which every treatment was randomly allocated with the different tanks (Table 1). 

The trial consisted of 3 treatment groups with 3 replicates per each. 

Table 1. Treatment definition 

Stocking a density of 900.000 Nauplii/tank. 

All treatments had the same Nauplii source (SIS Hawaii) and the same water exchange rate. 

Negative control (T1) was conducted as Conventional Current Practice without antibiotic, TIPTOPP POND and TIPTOPP AQUAG supplement. Water exchange will be performed to maintain quality. 

TIPTOPP Fermentation (T2) was conducted by applying fermented TIPTOPP POND at 2 Liter per m3 in one time before stocking, 1 Liter per m3 from N6 to Mysis3 and 2 Liter per m3 from PL1 to harvest. In addition, 50 gram of TIPTOPP AQUAG will be applied for each kg of artificial feed. 

TIPTOPP Direct use (T3) was conducted by directly applying TIPTOPP POND at 1 gram per m3 on daily basis. In addition, 50 gram if TIPTOPP AQUAG will be applied for each kg of artificial feed 

II. RESULTS 

1. Environmental conditions 

1.1. Water quality 

Water quality parameters were recorded during larval rearing period (day 0 through day 20) and presented as below. 

Table 2. Water quality parameters during the larval rearing period 

In general, all water quality parameters in this trial (Table 5) were kept in acceptable ranges for survival of larvae and post-larvae of Penaeus vannamei. 

1.2. Bacterial density in water samples 

Water samples were taken for total heterotrophic bacterial count (TBC) using PCA, TSA, TSA+, and TCBS media during the larval rearing period. 

a. Total bacteria count on PCA of water samples 

Results in Fig 5. indicated that enumeration of the total bacterial count (TBC) on PCA of water samples of different treatments has shown no significant differences among treatments from Days 0-5, Days 8-12 and Day 18. The TBC of T3 and T2 was significantly higher than the control on the remaining days of the trial (P<0.05). 

b. Yellow colony count on TCBS of water samples 

Results in Fig 6. indicated that most of the time the enumeration of yellow colony count (YCC) on TCBS of water samples of different treatments were no significant differences among treatments. 

c. Green colony count on TCBS of water samples 

Results in Fig 7. indicated that enumeration of the green colony count (GCC) on TCBS of water samples of different treatments were no significant differences among treatments. All groups have onlyGreen colonies were only appeared from Day 6 to Day 18 in all the groups. 

2. Larval health assessments & stress tests 

2.1. PLs performance

2.1.1. Survival rate 

Result in Fig 8. indicated that final survival rates of PLs of different treatments were no significant differences among treatments. 

The survival rates in treatment T1 and T3 were numerically higher than treatment T2.  

2.1.2. Larvae health evaluation 

PL10 samples were taken for larvae health evaluation at termination date of the larvae rearing and presented as below. 

Table 6. Summary results of post-larval quality assessent 

(Values are presented as mean (n = 3) ± standard deviation. The same letters on the same column were not significant differences (P > 0.05)) 

Result in Table 6 indicated that the PLs of  treatment T3 had higher quality than the control (T1) and treatment T2. 

2.2. Stress Tests

2.2.1. Salinity stress Test 

Result in Fig 9. indicated that survival rates of PLs in treatment T3 after salinity stress test was significant higher than treatment T2 (P>0.05). The methodology such as treatment T3 is seems to be mitigated the stress for the PLs in cases of salinity fluctuation. 

2.2.2. Formalin stress test 

Result in Fig 10. indicated that survival rates of PLs in treatment T3 and Control after formalin stress test were higher than treatment T2 (P>0.05).