formation of biofloc in fertilization pond at different salinities

In this research, Bio-Floc Growth in Fertilization Pond at Different Salinities was made in order to record the suitable salinity to bio-floc development and nutrient level to supply to

COLLEGE OF AQUACULTURE AND FISHERIES

FORMATION OF BIO-FLOC IN FERTILIZATION POND AT

DIFFERENT SALINITIES

TRAN HOANG CHIEN

A thesis submitted in partial fulfillment of the requirements for

The degree of Bachelor of Science for Aquaculture

Supervisor NGUYEN VAN HOA

COLLEGE OF AQUACULTURE AND FISHERIES

STUDY ON BIO-FLOC IN FERTILIZATION POND WITHOUT

ARTEMIA AT DIFFERENT SALINITIES

TRAN HOANG CHIEN

A thesis submitted in partial fulfillment of the requirements for

The degree of Bachelor of Science for Aquaculture

Supervisor NGUYEN VAN HOA

Can Tho, 01/2013

I sincerely thank Dr Nguyen Van Hoa dedicated to guide and help me in the process

of implementing the project

Thank you Mr Tran Huu Le, Mr Le Van Thong Thong and all my friends were enthusiastic help in the process of implementing this project

Today, raising Artemia biomass is very popular because it is an excellent food for aquaculture species Therefore, to enhance the production of Artemia biomass is always a hot topic e.g feed item, culture condition, culture system, etc

In this research, Bio-Floc Growth in Fertilization Pond at Different Salinities was made in order to record the suitable salinity to bio-floc development and nutrient level to supply to Artemia as a good natural food source

Research was conducted through experiments on the development of bio-floc at different salinity, there were 3 treatments in term of salinity difference (60‰, 80‰ and

100 ‰) and 3 replicate each Results showed that Bio-Floc contained highest value of crude protein at a salinity of 80‰ (21%) The suitable salinity of artemia is about 60‰ -100‰ (according toNguyen Van Hoa et al., 2007)

Acknowledgements 2

Abstract 4

Table of Contents 5

List of tables 6

List of figures 7

List of abbreviations 8

CHAPTER I INTRODUCTION 9

1.1 Introduction 9

1.2 Objectives 9

1.3 Studied contents 9

CHAPTER IILITERATURE REVIEW 10

A DISTRIBUTION 10

1 Classification 10

B Biological Characteristics 11

1 Habit 11

2 Feeding Characteristic 12

CHAPTER IIIMETHODOLOGY 13

3.1 Experimental Arrangement 13

3.2 parameters 13

3.3 data analysis method 16

CHAPTER IV RESULTS AND DISCUSSIONS 17

CHAPTER V CONCLUSIONS AND RECOMMENDATIONS 21

5.1 Conclusions 21

5.2 Recommendations 21

REFERENCE 22

APPENDIX 24

Table 5: pH level at 7am

List of figures

Figure 2: Artemia life cycle ( jumalon and et al,.1982) 11

Figure 6 Chart Shows the Development of the Bio-Floc at a Salinity of 60‰,

80‰, 100‰

18

List of abbreviations

VSS Volatile suspended solid

TSS Total suspended solid

CHAPTER I INTRODUCTION

1.1 Introduction

Bio-loc is defined as compose of diatom, macroalgae, exoskeleton, fecal pellet, remains of dead organism and bacteria a source of natural food rich in nutrition, bio-floc also have a role to improve water quality in ponds artemia

Aquaculture activities lead to a large amount of wastes, which has been carried into the environment (Fune-Smith and Briggs, 1998; Brune et al., 2003) Because of that, many researches and suggestions have been given to improve and protect the

environment Bio-floc technique has many advantages to solve the pollution problems and the environmental protection in aquaculture (Avnimelech, 2006; Hoang Tung, 2010) However, most of researches so far dealing with salinity around 30 ‰ or less as these are suitable for popular marine culture species (e.g tilapia, white leg shrimp)

There are not yet any researches or application on bio-floc that relates to culture Artemia in salt ponds where salinity is up to 80-120 ‰ (Nguyen Van Hoa et al., 2007) Normally, in Artemia pond culture, farmers applied about 2-5 tons of organic manure and 1-2 tons of supplemental feed (rice bran) for every crop (Nguyen Thi Ngoc Anh et al., 2009) per hectare every year, and thus with a thousand of hectare Artemia culture in the area, a huge amount of organic matter has been loaded into ponds every year Bio-floc is grown from this organic matter; its content is very high in nutrient value that is very useful for growth of aquatic organism

Therefore, research and application of bio-floc to ponds management play an

important role in cost decreasing and profit increasing Especially, it still gives apart of environmental protection and increases sustainability in culture

1.2 Objectives

 Recognizing the suitable salinity on the growth of bio-floc

 Recognizing the growth of bio-floc in saline water

 Recording nutrient level in bio-floc

1.3 Studied contents

 Formation and growth of bio_floc at saline water

 Influence of different salinity on formation and growth of bio-floc

CHAPTER II LITERATURE REVIEW

Species: Artemia, Leach (1819)

Depending on genetic characteristic and distribution that the scientists divide into some following six groups:

Artemia salina : Lymington (England, extinction)

Artemia tunisiana : Europ

Artemia franciscana : America (North, Central and South America)

Artemia persimilis : Achentina

Artemia urmiana : Iran

Artemia monica : Mono Lake, CA-USA

Figure.1: Map of Artemia Distribution (http://www.aquaculture.ugent.be/Education/coursematerial/online%20cours

- Salinity: 80-120 ‰

- Temperature: 22 - 35oC

- Dissolve oxygen: > 2 mg/l

- PH: 7.5 - 9.0

Figure 2: Artemia life cycle ((Jumalon and et al 1982)

Protein in feed can be absorbed by organism about 25-45%, and the remained amount stays on water environment under excessive food or waste of organism (according to National University of Ho Chi Minh City)

Besides bacteria, bio-floc also includes many other organisms such as fungi, algae; zooplankton…Bio-floc consists of living organisms about 2-20% and organic material about 60-70% Bio-floc not only plays an important role as a source of natural food but also improves water quality by metabolizing ammonia by heterotrophic bacteria

METHODOLOGY 3.1 Experimental Design

The experiment was carried out at Vinh Chau experimental station, Vinh chau

district, Soc Trang province There were 9 ponds with 150m2 /each (size) in the system; the experiment had 3 treatments with 3 replicates each

In the research, the rate of C: N was provided as 10:1 In Artemia ponds, heterotrophic bacteria can only grow well if the rate of C and N in the water is maintained at suitable ratio about 10/1 Supplying C helps the bacteria to develop, using all organic waste,

metabolizing ammonia, and cleaning environment

Chicken manure was used in the research to stimulate Bio-Floc formation Water level was 30 cm/pond Supplying manure in order to stimulate the water color (green water), of which a rate of 20kg/pond was used Raking to prevent lab-lab every day was necessary

Treatment 1: salinity of 60 ‰

Treatment 2: salinity of 80 ‰

Treatment 3: salinity of 100 ‰

POND T 3.1 POND T3.2 POND T3.3

POND T2.3 POND T2.2 POND T2.1

POND T1.1 POND T1.2 POND T1.3

Inlet

Figure 3.1 Diagram of position of ponds in Vinh Chau Experimental Farm Ocean

meter

- Turbidity was recorded every day

٭ Bio-floc sampling method:

- Sampling points were selected and marked randomly to sample during research period Samples were collected at every point that we marked, after that we transport them to Can Tho University for further analysis

٭ sample analyzing method:

- Weekly, every pond were sampled and put in 2 bottles (12/bottle) One bottle is used to analyze qualitative algae and another to analyze NO2, NO3, TOC, TVSS, and TSS

- Observing and recording the samples under microscope

- NO2- : colorimetric APHA et al., 1995

- NO3: cadmium removal APHA et al., 1995

- TSS, VSS analysis method:

Figure 4: water sampling

Figure 5: TSS and VSS analysis method

* Pond and brine preparation:

- Before the set-up, we prepared the ponds carefully and brine was prepared

about 1 month to let the water salinity increase up to the required salinity (i.e 60, 80 and 10 ‰)

- Pond management: to keep water level and salinity in sustainable

Filter paper after filting Water after filting

Clean filter paper

filting Water sample

Filter paper after filting To quantify TSS To quantify VSS

- Pond was observed to monitor the growth of Bio-Floc by recording water color

Bio-floc sample method:

- Bio-floc sampling to measure volume: 1 liter of water in the pond experiments on

a pyramid glass, after 20 minutes, recorded the volume of sediment to the bottom

of the cup

- Bio-floc sampling for nutrient analysis: in experimental ponds, water was pumped

into plastic barrels of 100 liters, let them 1 hour and collected sediment to the bottom of the barrel because the number of bio-floc enough to nutritional analysis

to reach 50 ml, so 100 liters of water was enough for analysis

3.3 data analysis method

- Using Excel spreadsheet to calculate average value, standard error and Statistical

Program to compare the mean among treatments, with the value p < 0.05)

RESULTS AND DISCUSSIONS

Environmental factor was remained in the research such as:

PH: during the culture period, pH varied and fluctuated between morning and in the

afternoon, the variation was recorded as in Table 1:

Table 1: pH fluctuation

(Note: Data with the same letters in column are not significant difference (p>0.05)

In the morning, pH fluctuates from 7.54 - 7.58 In the afternoon, pH fluctuated from 7.99 - 8.13 However, these differences were not statistically significant (p>0.05) except T1

These pH levels are suitable for Artemia growth, according to Nguyen Van Hoa et al (2007), Artemia Vinh Chau grow well on the pH level from 7 - 9

Table 2: Salinity (‰) fluctuation during research period:

During the experiment, salinity was maintained stable but the experimental period

Pond Morning (7 am) Afternoon (2 pm)

Table 3: The average volume (ml) of bio-loc obtained in all treatments at salinity of 60‰, 80‰, and 100‰:

At 100‰salinity, volume bio-floc obtained the highest in the first two days Results showed that bio-floc grow very fast in salinity 100‰ The next days the volume of bio-floc reduced due to deposition and death of algae

inoculation 17-Apr 22-Apr 29-Apr 5-May 7-May

60‰ 6.33±2.88 22.33±16.23 12.83±0.93 8.33±3.47 23.33±2.07

80‰ 8.83±2.70 24.33±16.06 18.00±10.16 9.83±2.54 24.50±3.49

100‰ 34.33±4.59 32.00±19.74 21.33±3.39 8.17±1.81 29.50±11.86

first eight days, at 60 ‰ and 80 ‰ bio-Floc increased continuously during the first 8 days, i.e from 6.33(ml) to 22.33(ml) and 8.83(ml) to 24.33(ml), respectively At the salinity of 60 ‰, 80‰ and 100 ‰ from day 8 to day 22 Bio-Floc decreases, the Biofloc was settled in the bottom and the algae density was decreased, and thus the bio-floc at the same time is decreased The research shows that bio-floc volume at the salinity of 100‰ dropped a lot after second, and then Bio-Floc increased again after day 22 because of algae development Bio-Floc developed very well in salinity 100 ‰ in time of 2-3 days of culture but in the next few days, there was a strong decline However, the volume of bio-floc obtained very high, therefore, salinity of 100 ‰ is suitable for Bio-Floc development

Table 4: The average protein obtained from samples in the research

Comparison of bio-floc volume obtained in this experiment, it was found that the volume

of bio-floc obtained in salinity 100‰ higher than the volume of bio-floc obtained at 80‰ However, based on the average amount of protein obtained, these results show that bio-floc engaged with highest protein value (10.90%) at salinity 80‰ Interestingly, this is also the suitable salinity for Artemia grows well in Vinh chau environmental conditions (Nguyen van Hoa et al., 2007)

Table 5: The average V(H2SO40.1N) obtained from samples in the research

Average

V (H2SO4 0.1 N)

Salinity of 60‰ 2.23±0.23 4.50±2.33 2.97±0.42 6.13±0.27 Salinity of 80‰ 3.07±0.46 4.20±0.31 5.93±3.02 7.50±0.79 Salinity of 100‰ 3.40±1.40 5.40±2.95 6.60±1.66 6.63±1.04

Salinity of 60 ‰ 7.52±0.44 7.95±1.27 9.01±1.24 9.58±1.10 Salinity of 80 ‰ 9.54±3.67 8.70±2.85 16.60±5.84 8.8±1.06 Salinity of 100 ‰ 6.87±0.88 9.08±1.37 15.7±3.05 8.60±0.33

Results showed that, at a salinity of 100‰, H2SO4 concentration increased

continuously from day 2 to day 15 and remained in high level until day 22 In the salinity

of 80‰, H2SO4 increased continuously from day 2 to day 22 this shows the salinity of 80‰, H2SO4 was born very much At the salinity of 100‰ and 80‰, bio-floc did grow very well, so H2SO4 concentration was also released in high level

CONCLUSIONS AND RECOMMENDATIONS

of bio-floc obtained very high

Based on the real farming conditions, in order to manage salinity at 100‰ is more difficult while bio-floc nutritional value is not high, thus encouraging farmers to maintain salinity 80‰ is the best

5.2 Recommendations

Research should be conducted in the fertilization ponds with the presence of Artemia

to appreciate the development and fecundity of Artemia when using bio-floc feed

Research should be conducted further experiments on the effects of temperature, pH and clarity to the development of bio-floc to accurately determine the impact of these conditions on the development of bio-floc

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