Effect of Methylene Blue and Effect of Methylene Blue and Sodium - - PowerPoint PPT Presentation

Effect of Methylene Blue and Effect of Methylene Blue and Sodium Chloride on the Bacterial Sodium Chloride on the Bacterial Load of Nile Tilapia ( Load of Nile Tilapia (Oreochromis Oreochromis niloticus niloticus) Fingerlings ) Fingerlings During Transport During Transport

Remedios B. Bolivar, Michael Angelo D. Remedios B. Bolivar, Michael Angelo D. Aragones Aragones and Gemerlyn G. Garcia and Gemerlyn G. Garcia*

*

College of Fisheries College of Fisheries-

• Freshwater Aquaculture Center and

Freshwater Aquaculture Center and

* *College of Veterinary Science and Medicine

College of Veterinary Science and Medicine Central Luzon State University, Science City of Mu Central Luzon State University, Science City of Muñ ñoz, Nueva Ecija

• z, Nueva Ecija

Philippines Philippines

Introduction Introduction

! ! Tilapia as a source of

Tilapia as a source of protein protein

! ! Contribution of tilapia to

Contribution of tilapia to Philippine aquaculture is Philippine aquaculture is 8.4% 8.4%

! ! Fingerlings production is

Fingerlings production is vital to the tilapia industry vital to the tilapia industry

! ! Survival of fingerlings

Survival of fingerlings during transport is critical during transport is critical

Significance of the Study Significance of the Study

! ! Use of methylene blue and sodium chloride

Use of methylene blue and sodium chloride in the reduction of bacterial load during in the reduction of bacterial load during fingerlings transport fingerlings transport

Statement of the Problem Statement of the Problem

! ! Maintenance of healthy tilapia fingerlings

Maintenance of healthy tilapia fingerlings for grow for grow-

• out
• ut

! ! Effective disinfectants in fish transport

Effective disinfectants in fish transport must be identified must be identified

Objectives of the Study Objectives of the Study

! !

To determine the effect of using two To determine the effect of using two concentrations of Sodium chloride and concentrations of Sodium chloride and Methylene blue on the bacterial load of the Methylene blue on the bacterial load of the transport water of Nile tilapia fingerlings transport water of Nile tilapia fingerlings

! !

To compare which concentration of the To compare which concentration of the two chemicals was more effective in the two chemicals was more effective in the reduction of bacterial load of the transport reduction of bacterial load of the transport water of Nile tilapia fingerlings water of Nile tilapia fingerlings

Bacterial Diseases of Fish Bacterial Diseases of Fish

! ! Columnaris disease

Columnaris disease

! ! Edwardsiella septicemia or Edwardsiellosis

Edwardsiella septicemia or Edwardsiellosis

! ! Vibriosis

Vibriosis

! ! Motile Aeromonad Septicemia

Motile Aeromonad Septicemia

! ! Pseudomonad Septicemia or Red Spot

Pseudomonad Septicemia or Red Spot disease disease

! ! Mycobacterios or Piscine Tuberculosis

Mycobacterios or Piscine Tuberculosis

Salt Salt

! ! Considered as “aspirin” of aquaculture

Considered as “aspirin” of aquaculture

! ! Commonly used to treat many external

Commonly used to treat many external parasites of fishes parasites of fishes

! ! Provides additional treatment in several

Provides additional treatment in several cases of bacterial disease of fishes cases of bacterial disease of fishes

Methylene blue Methylene blue

! ! It has inhibitory action on bacteria due to

It has inhibitory action on bacteria due to its binding effect with cytoplasmic structure its binding effect with cytoplasmic structure within the cell within the cell

! ! Reduces water mold infection

Reduces water mold infection

! ! Effective for treating external parasites in

Effective for treating external parasites in fishes fishes

Materials and Methods Materials and Methods

Treatments Treatments

Treatment Treatment Concentration Concentration

I I control control II II 1 g/l of sodium chloride 1 g/l of sodium chloride III III 2 g/l of sodium chloride 2 g/l of sodium chloride IV IV 3 mg/l methylene blue 3 mg/l methylene blue V V 1 mg/l of methylene blue 1 mg/l of methylene blue

Chemicals used in the study Chemicals used in the study

Salt Methylene blue

Test fish Test fish

! ! Size of fingerlings:

Size of fingerlings: size # 24 Nile tilapia size # 24 Nile tilapia

! ! Source of fingerlings:

Source of fingerlings: BFAR BFAR-

• NFFTC, CLSU

NFFTC, CLSU

Transport Procedure Transport Procedure

! ! Conditioning of fingerlings

Conditioning of fingerlings

! ! No feeding was done

No feeding was done before transport before transport

! ! Transport bags (20 x 30 x

Transport bags (20 x 30 x 0.003 in) 0.003 in)

! ! Loading rate of 1,100

Loading rate of 1,100 fingerlings/bag fingerlings/bag

! ! Duration of transport: 12

Duration of transport: 12 hours hours

! ! The bags were oxygenated

The bags were oxygenated

Collection of water samples Collection of water samples

! ! 100 ml of transport water was collected in

100 ml of transport water was collected in each bag each bag

! ! Samples of transport water were placed in

Samples of transport water were placed in test tubes for bacterial load and water test tubes for bacterial load and water quality analyses quality analyses

Bacterial counting Bacterial counting

! ! Serial dilution up to 10

Serial dilution up to 10-7 was done

! ! 0.01 ml from the bacterial suspension was

0.01 ml from the bacterial suspension was placed in petri plates and incubated for 24 placed in petri plates and incubated for 24 hours hours

! ! Number of Colony Forming Units per

Number of Colony Forming Units per milligram (CFU/ml) were counted milligram (CFU/ml) were counted

Data gathered Data gathered

! ! Bacterial count (CFU/ml)

Bacterial count (CFU/ml)

! ! Mortality (%)

Mortality (%)

! ! Water quality parameters

Water quality parameters

! ! pH

pH

! ! Temperature (

Temperature (o

• C

C) )

! ! D.O. (mg/L)

D.O. (mg/L)

Results Results

Bacterial count in the transport water at 0, 6 and 12 Bacterial count in the transport water at 0, 6 and 12 hours of transport of Nile tilapia fingerlings hours of transport of Nile tilapia fingerlings Bacterial count (CFU/ml) Bacterial count (CFU/ml) 4.6 x 10 4.6 x 108 c

8 c

3.7 x 10 3.7 x 109 c

9 c

4.7 x 10 4.7 x 109 c

9 c

V V 4.3 x 10 4.3 x 108 c

8 c

1.6 x 10 1.6 x 1010 c

10 c

3.3 x 10 3.3 x 1010 c

10 c

IV IV 2.1 x 10 2.1 x 108 b

8 b

3.5 x 10 3.5 x 108 b

8 b

4.5 x 10 4.5 x 108 b

8 b

III III 2.8 x 10 2.8 x 108 b

8 b

3.8 x 10 3.8 x 108 b

8 b

5.1 x 10 5.1 x 108 b

8 b

II II 5.5 x 10 5.5 x 1010 a

10 a

5.6 x 10 5.6 x 109 a

9 a

3.5 x 10 3.5 x 109 a

9 a

I I 12 12-

• hr

hr 6 6-

• hr

hr 0-

• hr

hr Treatment Treatment

Changes in bacterial count in the five treatments at Changes in bacterial count in the five treatments at every 6 every 6-

• hour intervals

hour intervals

0.00E+00 1.00E+10 2.00E+10 3.00E+10 4.00E+10 5.00E+10 6.00E+10

1 2 3 4 5

Treatment Bacterial count (CFU/ml)

Water quality parameters at 0, 6 and 12 hours of transport

15.4 15.4 15.4 15.4 14.3 14.3 14.5 14.5 15.0 15.0 12 12 14.1 14.1 14.7 14.7 14.9 14.9 14.4 14.4 14.7 14.7 6 6 18.2 18.2 18.6 18.6 18.5 18.5 18.2 18.2 18.6 18.6 Dissolved Dissolved

• xygen
• xygen

(mg/l) (mg/l) 30.7 30.7 30.3 30.3 30.3 30.3 30.2 30.2 30.4 30.4 12 12 29.1 29.1 29.1 29.1 29.5 29.5 29.3 29.3 29.4 29.4 6 6 25.3 25.3 25.8 25.8 25.5 25.5 25.4 25.4 25.2 25.2 Temperature Temperature ( (o

• C)

C) 7.6 7.6 7.7 7.7 7.7 7.7 7.8 7.8 7.7 7.7 12 12 7.5 7.5 7.7 7.7 7.5 7.5 7.3 7.3 7.7 7.7 6 6 8.1 8.1 8.2 8.2 7.7 7.7 7.6 7.6 8.0 8.0 pH pH 5 5 4 4 3 3 2 2 1 1 Treatment Treatment Duration Duration

• f
• f

transport transport (hour) (hour) Parameter Parameter

Mortality (number and percent) and percent survival

• f size # 24 Nile tilapia fingerlings after 12 hours of

transport 0.4 0.4 5 5 0.7 0.7 4 4 0.7 0.7 3 3 0.6 0.6 2 2 0.8 0.8 1 1 Mortality (%) Mortality (%) Treatment Treatment

Discussion Discussion

! ! Bacterial count in Treatment 1 was highest

Bacterial count in Treatment 1 was highest among all treatments at 0 to 12 hour of among all treatments at 0 to 12 hour of transport transport

! ! Bacterial count using methylene blue

Bacterial count using methylene blue showed highest decrease in bacterial count showed highest decrease in bacterial count compare to sodium chloride compare to sodium chloride

Conclusion Conclusion

! ! Methylene blue and sodium chloride were

Methylene blue and sodium chloride were both effective in reducing the bacterial load both effective in reducing the bacterial load in the transport water compared with the in the transport water compared with the control control

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