PAPER • OPEN ACCESS
Low Salinity Application to Improve Biofloc System in Early Grow-Out Stage of White Shrimp
(Litopenaeus vannamei)
To cite this article: M Syaichudin et al 2022 IOP Conf. Ser.: Earth Environ. Sci. 1118 012012
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Low Salinity Application to Improve Biofloc System in Early Grow-Out Stage of White Shrimp ( Litopenaeus vannamei )
M Syaichudin1*, Jumriadi2, A Gafur1, Akmal1, Rahmi3, Lideman1, Sadat 2, NM Juniyanto2, S Sujaka2
1 Research Centre of Fishery, Research Organization on Earth Sciences and Maritime, National Research and Innovation Agency, Cibinong, West Java Province, Indonesia. 16911
2 Brackishwater Aquaculture Development Centre, Takalar, South Sulawesi Province, Indonesia. 92254
3 Aquaculture Study Program, Faculty of Agriculture, University of Muhammadiyah Makassar, South Sulawesi Province, Indonesia. 90221
Abstract. Extreme time for the failure of white shrimp culture often occurs at the beginning of rearing, especially during disease attacks and environmental degradation, where high salinity is also a trigger. This research aimed to examine the robustness of shrimp culture with environmental design at low salinity at the beginning of rearing. Methodology: this study was conducted in two plastic ponds (900 m2), each with a seed stocking density (PL-8) of 250 indv/m2, where treatment A (control/initial salinity 27 ppt), while treatment B (low initial salinity 15 ppt) with rearing for 80 days. According to the results from both regimens, salinity changed from the beginning of stocking until the day of 43(27-28 ppt). There began to be similarities, where the addition of water always used normal seawater (33 ppt) until the end.
Vibrio harveyii disease attack in DOC 41 occurred in treatment A (control) which was marked by luminescence light from the water rearing, this was different in treatment B which was clean from the luminescence light of pond water. This is also thought to cause the survival rate of control to be lower. It appears that the survival rate (SR) calculation is different, whereas, in treatment A (control), it is only 86%, while in treatment B is 98%. The average body weight in treatment A averaged 8.19±1,36 g/indv, while treatment B averaged 8.69±1,55 g/indv. The results of the FCR calculation in treatment A (control) was around 1.57, while treatment B was 1.42 with total biomass on A 1.402 kg and for treatment B 1.624 kg. The implementation of a low salinity environmental design at the start of rearing can boost the white shrimp culture biofloc system's robustness, it can be concluded.
1. Introduction
Due to the decline in fish stocks brought on by predatory fishing, aquaculture is necessary to supply the demand for seafood [1]. The aquaculture industry's environmental impact grows as it expands rapidly, and the discharges from aquaculture damage the surrounding environment [2]. Although aquaculture is a sustainable seafood option, shrimp farming is frequently associated with eutrophication/water pollution, mangrove destruction/deforestation, disease transmission, high antibiotic consumption, and a decline in biodiversity in coastal regions, where these environmental problems result in financial losses [1, 3].
Producers employ bio-floc technology and commercial probiotics to improve water quality and be more productive in order to minimize the effects on shrimp farms. The biofloc technique offers a low-cost feed source and improved nutrient conservation. Compared to other methods, bio-floc technology has the advantage of being relatively inexpensive, making it a workable financial strategy for sustainable aquaculture [2].
The main problem in the cultivation of white shrimp (Litopenaeus vannamei) arises from various
diseases in the initial maintenance period (0-60 days). From global high demand for Pacific white shrimp (L. vannamei) has led to intensified cultivation and a wide range of disease problems, including bacterial diseases due to vibrios. According to a study done by [4], the shrimp aquaculture business has seen a few significant diseases that are primarily brought on by pathogenic bacteria and viruses. Over the past two decades, these illnesses have played a significant role in the decline of shrimp output. One of the worst illnesses in shrimp farming is linked to bacterial species, particularly those produced by Vibrio spp., which are thought to cause significant financial losses in shrimp farming across the globe [5].
Shrimp aquaculture sectors have been significantly influenced by infectious diseases, and most diseases in cultured shrimp are bacterial or viral in origin, with a few significant protozoal and fungal diseases [6]. Acute hepatopancreatic necrosis disease (AHPND), formerly known as early mortality syndrome (EMS), is a bacterial shrimp disease due to the action of PirABvp toxin secreted by Vibrio parahaemolyticus. AHPND can also be caused by other strains of Vibrios such as Vibrio harveyii, Vibrio punensis, Vibrio owensii, Vibrio campbellii, and Vibrio alginolyticus which possess pVA1-type plasmid where the genes encoding the toxin are located [7]. V. alginolyticus is an opportunistic pathogen of shrimp known to cause mortalities under poor environmental conditions [8]. Vibrio harveyii, which causes luminous vibriosis, is another etiological agent for mass mortalities in Penaeid shrimp [9]. The two most significant viral diseases affecting shrimp farming are the White Spot Syndrome Virus (WSSV) and Infectious Myonecrosis Virus (IMNV). The highly pathogenic WSSV kills shrimp completely three to ten days after the onset of clinical signs.
Vibriosis disease is typically thought to result from opportunistic infection in shrimp with weakened immune systems, harsh environmental factors, and high bacterial concentration in the water column. The results of studies conducted by [10] have suggested that Vibrio spp. White shrimp L. vannamei experience mass extinction when there are concentrations in the water equivalent to or higher than 104 CFU/ml. Additionally, there is a growing understanding that factors including water temperature, pH, salinity, and nutrients in the water column have an impact on the prevalence of Vibrio spp. [11].
According to [12], the Vibrio genus expands quickly when the aquatic environment is improved by the buildup of organic waste.
According to certain research, AHPND Vibrio is more pathogenic in high-salinity environments than in low-salinity environments. According to research done by [13], less survival was seen in infected and uninfected shrimp kept at salinities of 28 ppt and 10 ppt as opposed to those at 20 ppt. The maximum survival rate was seen in shrimp maintained at 20 ppt (78.70%), followed by those at ten ppt (69.45%), among the infected group. Shrimp survival was noticeably low in those maintained at 28 ppt (54.89%);
The mortality of uninfected shrimp maintained at various salinities did not differ significantly. The mechanism might explain how salinity affects P. monodon infection with VPAHPND. When shrimp are stressed and exposed to greater salinities [14], V. parahaemolyticus harmful genes are more effectively produced.
When moved to environments with high or low salinity levels, P. monodon loses some of its immunological capacity and disease resistance [15]. The observed mortalities in the present study demonstrate that deaths from VPAHPND appear to be age-related, supporting arguments that excessive salt is a risk factor for the disease. Shrimp at DOC 150 were not affected by exposure to concentrations as high as 107 CFU/mL, but younger shrimp subjected to concentrations of 106 and 107 CFU/mL had significant mortality. The survival of infected shrimp in the two age groups demonstrates that as shrimp get older, their susceptibility to VPAHPND decreases in the absence of a stressor. The results supported [16], who reported 40–60% mortality rates for penaeids at DOC 46 and 96, respectively.
Low salinity (20 ppt) water sources appear to lower the prevalence of the illness. Peak incidence seems to occur from April to July during the hot and dry seasons [17]. [18] explored the correlation between salinity and AHPND in P. vannamei. Vp AHPND broth has challenged pathogen-free shrimp cultures (5, 10, 15, and 20 g L−1 of NaCl). Histological damage and the detection of pirABVp toxin genes by PCR analysis showed that Vp AHPND infected shrimp in all salinity treatments. But cumulative mortality showed a higher survival rate in shrimp kept at lower salinities. Given that P. vannamei reproduces more efficiently in high-saline environments, it is likely that more pirABVp toxin was created, leading to increased cumulative mortality in P. vannamei when maintained in these conditions [19].
An effort must be made to maintain the stability of the maintenance environment to reduce stress levels and stop the spread of epidemic illnesses. One such attempt involves the use of biofloc technology.
Sustainable aquaculture development is necessary to safeguard the environment and natural resources.
Due to its sustainability, the method for raising aquatic organisms known as "Biofloc Technology" (BFT) has recently gained prominence. Maintaining a healthy carbon-nitrogen balance, recycling nutrients, and increasing natural production in the farming system, this practice leads to improvements in water quality [20]. Heterotrophic bacteria can make protein from organic carbon, and ammonia gains from absorbing ammonia nitrogen when the balance of carbon and nitrogen (C: N) is between 15-20:1. [21].[22] claims that when raising Litopenaeus vannamei in BFT systems, the inclusion of organic carbon sources, like sugar cane molasses, can limit rises in the concentration of total ammoniacal nitrogen. Bacteria and other microbes use carbohydrates (sugars, starch, and cellulose) as food to generate energy, and to grow, i.e., to produce proteins and new cells. The reduction of inorganic nitrogen accumulation in ponds is based on carbon metabolism and nitrogen-immobilizing microbial activities. [21]. As a waste product from tapioca flour production, cassava dregs can be supplied in considerable amounts and sustainably as a molasses substitute. It is necessary to undertake fermentation to make the carbohydrate structure of cassava heaps simpler for fish to digest [23].
The growth rates and bacterial biomass output of heterotrophic bacteria are ten times higher than those of nitrifying bacteria. Autotrophic bacteria carry out nitrification by converting ammonia to nitrite and nitrate. Nitrite tends to build up at substantial concentrations in rearing conditions without water renewal because of the sluggish development of these bacteria [24]. Because heterotrophic bacteria use organic carbon from molasses as an energy source and present faster growth than nitrifying bacteria, they eventually reduce the nitrification rate in the system [25]. [26] reported a reduction of 50 % in the nitrification rate when the C/N ratio was increased from 0 to 1. However, they did not verify any difference in the nitrification rate between C/N ratios of 1 and 2, thus confirming the negative impact of organic matter on the nitrification rate.
Compared to white shrimp culture without biofloc, biofloc technology can boost productivity, lower feed consumption, stabilize conditions, and maintain shrimp health in intensive systems of Litopenaeus vannamei [27]. According to some information, white shrimp (Litopenaeus vannamei) can consume biofloc [28]. The microbial floc produced, according to [29], comprises a variety of nutrients, including protein (19%-32%), fat (17%-39%), carbs (27%-59%), and ash (2%-7%) that is suitable for the growth of vaname shrimp. It is essential to enhance waste management in intensive shrimp ponds since the technology for intensive shrimp culture is evolving quickly, producing aquaculture waste that could harm the aquatic environment. In extensive shrimp pond sewage treatment facilities, sedimentation tanks can assist reduce waste organic matter. In contrast, liquid waste that goes through the bioremediation route is transformed into simple nutrients for the growth of phytoplankton and macroalgae before being returned to the seas [30].
Environmental management is improved to reduce the level of disease outbreaks at the beginning of maintenance, especially vibriosis disease (AHPND/EMS); an ecological design test is carried out by reducing the initial salinity of care. This maintenance is carried out to strengthen or strengthen the cultivation of white vannamei shrimp (L. vannamei) reared in plastic ponds using a bio-floc system compared to the control without reducing salinity (standard).
2.Materials and Methods 2.1. Research Preparation
This research about the environmental design with low salinity at the beginning of rearing as an effort to improve the robustness of white shrimp (L vannamei) culture by the bio-floc system was carried out at the White Shrimp Production Installation organized by Brackishwater Aquaculture Development Centre (BADC Takalar) at Mappakalompo Village, Galesong District, Takalar Regency, South Sulawesi. The equipment used in this study included two plastic ponds with a combined area of about 900 m2, a root blower with a 3 HP motor, 12 paddle wheels with a 1 HP motor, a cast net, a bottom harvest net, a probiotic culture container, a sampling bottle, an Imhoff cone, and tools for measuring the water quality.
At the same time, the ingredients include molasses, yeast, PL8 seeds (domestic white shrimp), shrimp feed (D0-PV2), probiotics (bacillus and lactobacillus), lime, and vitamin C.
2.2. Research Design
This research employs an environmental design that applies low salinity at the beginning of rearing to increase the robustness of white shrimp (L. vannamei) cultivation via a biofloc system. The research activity will compare Treatment A (the control) and Treatment B (the standard initial salinity treatment)
both of which had initial salinities of 15 ppt and 27 ppt, respectively, at the first rearing. The addition of water exchange always used regular seawater with a salinity of around 33 ppt. The stocking density of each treatment for this research was 250 indv/m2, with the cultivation period being carried out for 80 days.
Monitoring and observation activities on shrimp health and maintenance of water quality level from the beginning of rearing to the end. Feed management in the initial 30 days of rearing applies a blind feeding system, then the level of feeding (Feeding Rate) is based on the average body weight from the day 31 until the end of the research. Artificial feed was administered four times each day, where feeding took place at 6:00 a.m, 11:00 a.m, 18:00 p.m, and 22:00 p.m.
2.3.Parameters Measurement
The parameters measured in this research activity, such as 1). Shrimp health: growth performance:
average body weight (ABW), average daily growth (ADG), and survival rate (SR), 2). Measurements of biofloc level were made using an Imhoff cone and the quality of biofloc using microscopic analysis, 3).
Productivity: total biomass and Feed Conversion Ratio (FCR), 4). Water quality parameters: pH, temperature, salinity, alkalinity, dissolved oxygen, and ammonia. The measurements of water quality for temperature (thermometer), salinity (refractometer), pH (pH meter), and DO (dissolved oxygen meter) were carried out in situ, but ammonia was analyzed in the laboratory using a spectrophotometer.
Parameter observation for productivity that is collected uses this formula. Average ABW is the average weight of shrimp (gr/shrimp) from the sampling results. It can be calculated by this formula:
ABW = shrimp weight/number of shrimp. FCR is the amount of feed given to produce one kg of fish meat. FCR calculation used is as follows pattern: FCR = F / (BT + BD) – B0, where F = total feed (kg), BT
= total biomass final (kg), BD = dead fish biomass (kg), and B0 = total initial biomass (kg). SR is the survival of shrimp compared to the number of stockings and expressed by percent. SR can be calculated by the following formula: SR = (Nt /N0) x 100%, where Nt = the final number of shrimp and
N0 = the initial number of shrimps.
2.4. Data Analysis
All data collected during this research were analyzed by descriptive statistics and then presented using charts, such as water quality, biofloc performance, shrimp growth, and survival rate, while data on productivity were presented in graph and table form.
3. Results and Discussion
3.1. Observation of The Maintenance of Water Salinity Dynamics
Based on the experiment result, the dynamics of salinity (Figure 1) showed that at the first treatment A (control) at 27 ppt and for treatment B at 15 ppt. The salinity of treatment B gradually increases coincide with water exchange from the reservoir (33 ppt), then began to be similar on the day of culture (DOC) 42nd (27-28 ppt). Water salinity keep increasing at the same time to the end of the research that achieved 33-34 ppt.
Figure 1. The dynamics of water salinity at the experiment pond 35
30 25 20 15 10
33 33 34
32 33 34
27 28
26 25 33 34
32 32 33
29 27 23
Treatmen A (Salinity - ppt) Treatmen B (Salinity - ppt)
15 16
2 7 28 35 42 58 64 70 77 80
Day of Culture / DOC (day)
Salinity (ppt)
The problems of culture that arise in the first 40 days of culture, with the emergence of various shrimp diseases, especially Acute Hepatopancreatic Necrosis Disease (AHPND) or Early Mortality Syndrome (EMS) diseases that caused by Vibrio parahaemolyticus, and since there is information that the virulence level of the disease decreases at low salinity, need to carry out by maintaining with low salinity at the beginning of rearing. According to [13]'s research, the survival rate of AHPND-infected and uninfected shrimp maintained at salinities of 28 ppt, and ten ppt was observed to be lower than that of shrimp held at 20 ppt. Among the infected group, the highest survival rate was seen in shrimp maintained at 20 ppt (78.70%), followed by those at ten ppt (69.45%).
According to the research done by [31], Litopenaeus vannamei juveniles and postlarvae can be successfully cultivated at low salinity levels of 4 ppt, where there are no changes in growth or survival rates between this level and 30 ppt. Salinities below four ppt showed a detrimental effect on postlarval development and survival. Juveniles, however, were able to endure at both two ppt and 30 ppt.
Shrimp are well adapted to low salinities, according to glucose and protein values found in their hemolymph throughout the study [32]. (1, 10, and 15 ppt). Although only 83.3% of shrimp raised at salinity ten ppt survived, shrimp at this salinity showed greater glucose concentrations in hemolymph at the start and conclusion of the trial.
3.2. Observation of The Water Microbiology Dynamics
Based on the experiment result, the dynamics of bacteria (Figure 2) showed that on the day of culture 01- 28th total bacteria for treatment B quickly increased compared with treatment A (control). After slightly decreasing, this treatment B condition continued to highly increase on the day of culture (DOC) 42-64th to get the top of total bacteria (3,95x104 CFU). Observation of total vibrio showed from the first DOC until 58th at treatment A (control) highly if total vibrio compares with treatment B.
Vibrio spp. infections, which have been causing significant losses and mass mortality in intensive shrimp farming, have been a major problem. One of the pathogenic organisms, Luminescent V. harveyiii, can infect penaeid shrimp at several phases of development, including early larvae (Nauplius, Zoea, and Mysis), postlarvae, juveniles, and grow-outs [33]. The primary pathogen in marine fish and crustaceans is luminescent Vibrio harveyii [34].
Figure 2. The dynamics of total bacteria and total vibrio at the experiment pond
Based on the experiment result, the dynamics of the ratio of total bacteria and total vibrio (Figure 3) showed that at the day of culture (DOC) 2nd - 58th at treatment B more constant with a ratio average of 0,02 – 0,19 if compare treatment A (control) with ratio average 0,03 – 0,43. At DOC 64th both treatments A and B same at the highest ratio around 0,40-0,42 then quickly decreased at around 0,19 – 0,21 (DOC 77th).
40000 35000 30000 25000 20000 15000 10000 5000 0
Treatmen A (Total Bacteria) Treatmen B (Total Bacteria) Treatmen A (Total Vibrio) Treatmen B (Total Vibrio)
39.500 39.000
28.500
15.900
8150 9.750
6.250 6.680 7.100
1600
2 7 28 35 42 58 64 70 77 80 Day of Culture / DOC (day)
Total Plate Count (Bacteria) CFU
Figure 3. The dynamics of total bacteria and total vibrio ratio at the experiment pond
Observation of the luminous vibriosis disease with the visual performance of water luminescence in ponds showed in Table 1 below. From the first day of culture until the 40th both of treatments none of luminescence light appears in the experiment ponds but starting from DOC 41st at treatment A (control pond) appears slight luminescence light for 3 days then moderate luminescence light for 2 days, since this day was done treatment to overcome it. The treatment had been done that are water exchange and the application of garlic on the feed. The peak or heavy luminescence light occur since on the day of culture 45th for three days, while treating for this luminescence light disease, the luminescence light gradually decreased from moderate to slight, and none of the luminescence light for 12-15 days.
Table 1. Observation data of luminous vibriosis in experiment ponds No
Day of Culture / DOC (day)
Luminous Vibriosis in Ponds
Treatment A Control / normal initial salinity 27 ppt
Treatment B Low initial salinity 15 ppt
1 40 None of the Luminescence Light None of the Luminescence Light 2 41 Slight Luminescence Light None of the Luminescence Light 3 42 Slight Luminescence Light None of the Luminescence Light 4 43 Moderate Luminescence Light None of the Luminescence Light 5 44 Moderate Luminescence Light None of the Luminescence Light 6 45 Heavy Luminescence Light None of the Luminescence Light 7 46 Heavy Luminescence Light None of the Luminescence Light 8 47 Heavy Luminescence Light None of the Luminescence Light 9 48 Moderate Luminescence Light None of the Luminescence Light 10 49 Moderate Luminescence Light None of the Luminescence Light 11 50 Moderate Luminescence Light None of the Luminescence Light 12 51 Slight Luminescence Light None of the Luminescence Light 13 52 Slight Luminescence Light None of the Luminescence Light 14 53 Slight Luminescence Light None of the Luminescence Light 15 54 None of the Luminescence Light None of the Luminescence Light
0,50 0,45 0,40 0,35 0,30 0,25 0,20 0,15
Treatmen A (Ratio Total Vibrio/Total Bacteria) Treatmen B (Ratio Total Vibrio/Total Bacteria)
0,43 0,40 0,42 0,43
0,35 0,34
0,28
0,19 0,19
0,21
0,12 0,23
0,19 0,10 0,03
0,05 0,00
2
0,18
0,11 0,09
0,02 0,09 0,06
7 28 35 42 58 64 70 77 80
Day of Culture / DOC (day)
Ratio Tot. Bac / Tot. Vibrio
Based on the total vibrio data from the experiment the to analyze of percentage ratio of yellow and green vibrio for treatment A (control pond) at the beginning of culture, there is a high percentage of green vibrio (75%), then decreases until the day of culture 35th. The problem was from DOC 42nd, a high percentage of green vibrio was coming (83%) until DOC 58th, this condition coincides with the appearance of luminescence light diseases at DOC DOC 41st that were caused by Vibrio harveyii. Good progress for a high percentage of yellow vibrio from the day of culture 64th until the end of the experiment. But analyzing the percentage ratio of yellow and green vibrio for treatment B at the beginning of culture, there is a high percentage of green vibrio (94%), then highly decrease on the day of culture 7th, and for the short term increase again on DOC 28th. Unfortunately, from the day of culture 35Th, the percentage of green vibrio until the end of the experiment is still on the low level of green vibrio percentage, even none of the green vibrio at DOC 64th. DOC 80th.
Immunomodulators are now widely utilized and studied to improve fish immunity worldwide.
Probiotics, prebiotics, and synbiotics have become familiar to boost fish resistance to vibriosis. They have been approved as viable substitutes for the current practice of limiting antibiotic use in aquaculture, resulting in decreased mortality and improved health and welfare of aquatic organisms. Numerous studies hypothesized that probiotics, prebiotics, and synbiotics could be employed as food or water additives to boost immunity and decrease the mortality brought on by various fish infections like Vibrio species [35].
3.3. Observation of Shrimp Growth and Productivity
Based on the observation of growth performance during the 80th day rearing period that is shown in Figure 4 below, it appears that both treatments at the first crop until the day of culture (DOC) 18 average body weight tend to be same, just still in 0,11g/indv. It’s very low, we use local or domestic seeds that tend to have disease resistance, not fast-growth seeds. Then the average body weight gradually increased until at the end of the experiment, it seems at treatment A gets ABW around 8,19 g/indv while treatment B is slightly higher with an average of 8,70g/indv. Recycling nutrients that enter the crop system is the primary concept of biofloc technology (Biofloc Technology/BFT), which makes their utilization more effective. Excess nutrients derived from the excretions of cultivated organisms and feces and feed residues that are not consumed will be converted by heterotrophic bacteria into bacterial biomass so that water quality is maintained and bacterial biomass which then forms flocs that can be consumed by cultivated organisms [29].
Figure 4. Average body weight (ABW) obtained from the experiment treatments A and B The results of monitoring the average daily growth (ADG) are shown in Figure 5 below, it appears that both treatments at the first experiment crop until the day of culture (DOC) 28 average daily growth tend to same, then until at the end of research in treatment A and B slightly fluctuating from 0,04-0,40 g/day.
Treatment B Low initial salinity 15 ppt
Treatment A Control / normal initial salinity 27 ppt
8,70 8,05 7,18 5,93 5,16 3,91 3,01 3,27 2,04 0,010,010,030,080,110,32
0,75 1,141,39 5,617,207,43
8,19 5,23
1,99 3,073,364,09
1 7 11 14 18 21 25 28 30 35 39 42 45 51 56 60 66 80 Day of Culture / DOC (day)
ABW Shrimp (g/ind)
Figure 5. Average daily growth (ADG) obtained from the experiment treatments A and B Table 2 below showed that at the end of the harvest that was preceded by two partial harvests, the total harvest was carried out on the day of culture (DOC) 80 days obtained shrimp biomass in treatment A was 1.402 kg, while for treatment B slightly better it was around 1.624 kg, with a feed conversion ratio (FCR) respectively for treatment A around 1,57 and treatment B 1,42. The survival rate (SR) at the end of the experiment for treatment A obtained 86%, while treatment B was better with a value of 98%. These results from treatment B (survival rate and FCR) were better than the results that had been done by [36] also concerned about the rearing of white shrimp (L. vannamei) in low salinity (10 ppt) with different stocking densities from 175 indv/m2 – 275 indv/m2 at circle tarpaulin pond for 60 days rearing had given data about Survival Rate (SR) from 55.03% - 91,39% and the best Feed Conversion Ratio (FCR) was 1.6.
An experiment about the productivity performance of the domestic vannamei shrimp in an intensive system with the application of biofloc is quite feasible and provides a good profit for the sustainability of aquaculture [37], with 100 days of rearing, getting a survival rate of 75.3% and a feed conversion value (FCR) of around 1.48.
Table 2. Data of the pond productivity from the experiment in treatment A and treatment B.
Research Treatment
No. Description Unit Treatment A Control
initial salinity 27 ppt
Treatment B Low initial salinity 15 ppt
1 Ponds Area m2 900 900
2 Number of PL Stock individu 225.000 225.000
3 Stocking Density indv/m2 250 250
4 Day Of Culture Days 80 80
5 Survival Rate % 86 98
6 Population Individu 193.476 221.592
7 Average Body Weight g/indv 8,19±1,36 8,69±1,55
8 Size indv/kg 122 115
9 Total Biomass kg 1402 1624
10 Total Feed kg 2.200 2.300
11 FCR Ratio 1,57 1,42
0,45 0,4 0,35 0,3 0,25 0,2 0,15 0,1 0,05 0
Treatment A Control/normal initial salinity 27 ppt Treatment B Low initial salinity 15 ppt
0,40
0,27 0,31
0,24 0,24
0,18 0,21
0,13 0,11
0,11 0,13 0,07
0,19 0,15
0,13 0,15
0,06 0,05
0 0,00 0,01 0,02 0,10 0,09 0,08
0,01 0,04
1 7 11 14 18 21 25 28 30 35 39 42 45 51 56 60 66 80 Day of Culture / DOC (Day)
ADG Shrimp (g/day)
5 4,5 4 3,5 3 2,5 2 1,5 1 0,5 0
Treatment A Control / normal initial salinity 27 ppt
Treatment B Low initial salinity 15 ppt
2
4,5 4 3,1
2 1,6 1,5 1,4
2,3 1,6
2,2 1,7 2
1 1,1
1,5
0,1 0,2 1,3 1,1 1,3
14 20 28 0,1
31 40 47 50 54 62 70 77 80 Day of Culture / DOC (day)
According to research cited in [13], white shrimp (L. vannamei) infected with the AHPND have a greater survival rate when raised in salinities of 10 and 20 ppt as compared to 28 ppt. The bacterial community at 28 days was dominated by Proteobacteria, Bacteroidetes, Planctomycetes, Chlamydiae, and Firmicutes, where to get a proportion of 81% inferring KEGG functions of this bacterial community is associated with metabolism. This consisted of the result of rearing L. vannamei postlarvae in a bio- floc system in low salinity media 5.0 ppt. The observations offer a fresh perspective on how the bacterial population functions in the biofloc system, while L. vannamei PL is being maintained at low salinity [38].
Fish farming operations and the aquaculture sector may be very interested in the positive effects of synbiotics. However, several parameters, such as the interactions between the probiotic strain and prebiotic substance present in the synbiotic product, should be considered when choosing a "synbiont."
For the usage of synbiont in aquaculture to be established, there must be synergism or synergistic interaction. The researcher should conduct additional research investigations to ascertain the right relationship and processes of synbiotics on fish's immunity and disease resistance against the challenge of pathogenic microorganisms [35].
3.4. Observation of The Biofloc Formation Dynamics
Based on the observation of biofloc formation dynamics at the experiment ponds in Figure 6 below, both treatments showed that at the beginning of biofloc formation on the day of crop 14th to 20th at around 0,1 ml/L, then for treatment B formation of biofloc gradually increase until at the end of experiment reach 4.5 ml/L. According to [39], the typical volume of floc is 2-4 ml/l by observation using an Imhoff cone. This is different from treatment A (control) where biofloc formation from the day of crop 40th to 54th gradually decreased coinciding appearance of luminescence light caused by Vibrio harveyiii and the treatment program with water exchange 20-25% daily until none of the luminescence light at the experiment pond A.
The biofloc formation process involves the earliest phases of raising marine shrimp utilizing biofloc technology (BFT). Total suspended solids are increasing simultaneously, and alkalinity and pH levels are falling. The alkalinity and pH are reduced by autotrophic bacteria in biofilms and bio-floc as they consume inorganic carbon [40].
Figure 6. Observation of the biofloc formation dynamics at the experiment ponds
Observations of the biofloc at the beginning of growth the biofloc particles were small andtransparent and increased with the increase in the maintenance period [37]. [41] claims that for every gram of ammonia nitrogen transformed into microbial biomass, 8.07 g of microbial biomass and 9.65 g of carbon dioxide are produced, whereas 4.71 g of dissolved oxygen, 3.57 g of alkalinity, and 15.17 g of carbohydrates are consumed. Considering the possible stratification of alkalinity and pH in biofilms, alkalinity higher than 200 mgCaCO3/L is recommended [42], especially when the water renewal rate is minimal or nonexistent, as in BFT systems. There is no optimal amount of alkalinity, and in systems with limited water exchange, the only alkalinity values accessible are recommended levels. There must be between 100 and 150 mg CaCO3/L of alkalinity. However, [43] shows that alkalinity concentrations
Biofloc Volume (ml/L)
8,2 8,0 7,8 7,6 7,4 7,2 7,0
8,0 8,1
7,9 8,0
7,7
Treatment A Control initial salinity 27 ppt Treatment B Low initial salinity 15 ppt 7,9
7,5 7,6 7,6
7,6 7,6
7,6 7,4 7,5
7,4 7,2
7,4 7,4 7,5
7,2
7,2 7,0
1 7 14 21 28 35 42 58 64 70 77 80 Day of Culture / DOC (Day)
below 100 mg CaCO3/L and pH concentrations below seven negatively affect the nitrification rate in BFT systems.
3.5. Observation of The Water Quality Parameters
The observation of water quality parameters in this experiment for 80 days of rearing covers some parameters such as pH, alkalinity, dissolved oxygen, water salinity, ammonia, nitrite, and phosphate.
Water quality management and biosecurity protocols are frequently used to treat vibriosis. [35]. In observing the pH parameters of the water during research, as shown in Figure 7 below, both treatments at the beginning of the experiment were slightly higher (7,6-8,1) and then gradually decreased until the end of the research (7,0 – 7,2).
The biofloc formation process involves the earliest phases of raising marine shrimp utilizing biofloc technology (BFT). Total suspended solids levels are rising simultaneously while alkalinity and pH levels are falling. The autotrophic bacteria found in biofilms and bio-floc consume inorganic carbon, which results in the reduction of alkalinity and pH parameters [40]
Figure 7. Observation of the dynamics of pH parameters at the experiment ponds
Observation of the water alkalinity parameters during the experiment can be seen the Figure 8 below, at the beginning of the culture both treatments gradually increased until the day of culture 35th (128,96- 152,52). For treatment A (control) at DOC 42nd which coincides with the appearance of luminescence light with water exchange treatment, the value of alkalinity highly increased based on the water added from the reservoir to reduce Vibrio harveyiii in pond A. After a day of crop 58th, both of treatments showed that alkalinity gradually decreased until at the end of the experiment.
Figure 8. Observation of the dynamics of alkalinity parameter at the experiment ponds The experiment done by [40] showed that higher alkalinity (150-300 mg CaCO3/L) favors biofloc formation and the establishment of nitrifying bacteria, whereas low-level alkalinity (75 CaCO3/L). The study presented the highest levels of ammonia and nitrite compared to the 150 and 300 treatments due to the nitrifying and heterotrophic bacteria that cause the bio floc to absorb inorganic carbon, the alkalinity is reduced (41).
180 160 140 120 100 80 60 40
156,61 149,2 152,52 131,83 129,58 116,88
102,18 109,96
133,75 140,28 139,93 129,68
140,28 137,17122,41 108,74
128,96 124,62
113,53 106,04 95,86
81,63 86,87 Treatment A Control initial salinity 27 ppt
Treatment B Low initial salinity
81,53
15 ppt
1 7 14 21 28 35 42 58 64 70 77 80 Day of Culture / DOC (day)
Alkalinity (mg/l) Acidity / pH
Figure 9. Observation of the dynamics of dissolved oxygen parameters at the experiment ponds The results of the observation of dissolved oxygen parameters during the experiment showed that in Figure 9 above, both treatment ponds from the beginning of culture until the end of the experiment, are in the range of optimum for normal rearing, at around 5,81 – 8,54 mg/l. According to [41], 8.07 g of microbial biomass and 9.65 g of carbon dioxide are created for every gram of ammonia nitrogen transformed in microbial biomass, along with 4.71 g of dissolved oxygen, 3.57 g of alkalinity, and 15.17 g of carbohydrates.
Observation of water temperature parameters during the experiment, both treatments at around 270- 280C. This condition was quite optimal. The investigation was reported by [44]. Higher temperature is a risk factor for AHPND, as evidenced by the more significant mortality of infected shrimp kept at higher temperatures. This might result from the disease and shrimp being affected by high temperatures in concert. Penaeids show effects at higher temperatures; in addition, most bacteria's hazardous gene becomes more virulent as temperature rises.
The level of ammonia content during rearing periods in treatment A slightly higher compared to treatment B, while the level of nitrite content in treatment A (41,444-47,268 mg/l) was higher compared to treatment B (21,906-25,345 mg/l), but for nitrate level on treatment B (2,001-2,144 mg/l) slightly higher than treatment A (1,584-1,641 mg/l). To convert one gram of total ammoniacal nitrogen (TAN) into nitrate, roughly 4.18 grams of oxygen, 7.07 grams of alkalinity, and 0.17 grams of bacterial biomass are used up [42].
4.Conclusion
Based on the results of the experiment can be concluded that in both treatments, there were fluctuations of salinity at the beginning of stocking up to the day of 43 (27-28 ppt), then there began to be similarities, where the addition of water always used normal seawater (33 ppt) until the end. Vibrio harveyii disease attack in DOC 41 occurred in treatment A (control) which was marked by luminescence light from the water rearing, this was different in treatment B which was clean from the luminescence light of pond water. This is also thought to cause the survival rate of control to be lower. It appears, that the calculation of Survival Rate (SR) is different, where in treatment A (control) it is only 86%, while in treatment B is 98%. The average body weight in treatment A averaged 8.19±1,36 g/indv while treatment B averaged 8.69±1,55 g/indv. The results of the FCR calculation in treatment A (control) was around 1.57, while treatment B was 1.42 with total biomass on A 1.402 kg and for treatment B 1.624 kg. The conclusion can be drawn that the application of an environmental design with low salinity at the beginning of rearing can increase the robustness of the white shrimp culture bio-floc system.
Acknowledgments
Grateful thanks addressed to my colleagues and technician for the assistance and encouragement in conducting the research and in preparing this article.
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