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 ( Oreochromis Oreochromis Load of Nile Tilapia ( niloticus ) Fingerlings ) Fingerlings niloticus During Transport During Transport Remedios B. Bolivar, Michael Angelo D. Aragones Aragones Remedios B. Bolivar, Michael Angelo D. * and Gemerlyn G. Garcia * and Gemerlyn G. Garcia College of Fisheries- -Freshwater Aquaculture Center and Freshwater Aquaculture Center and College of Fisheries * College of Veterinary Science and Medicine * College of Veterinary Science and Medicine Central Luzon State University, Science City of Muñ ñoz, Nueva Ecija oz, Nueva Ecija Central Luzon State University, Science City of Mu 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- -out out for grow ! 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 Concentration Treatment Concentration I control I control II 1 g/l of sodium chloride II 1 g/l of sodium chloride III 2 g/l of sodium chloride III 2 g/l of sodium chloride IV 3 mg/l methylene blue IV 3 mg/l methylene blue V 1 mg/l of methylene blue V 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- -NFFTC, CLSU NFFTC, CLSU BFAR

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 -7 was done ! Serial dilution up to 10 ! ! 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 ! o C ! Temperature ( Temperature ( o 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 Treatment Bacterial count (CFU/ml) Treatment Bacterial count (CFU/ml) 0- -hr hr 6- -hr hr 12- -hr hr 0 6 12 9 a 9 a 10 a 3.5 x 10 9 a 5.6 x 10 9 a 5.5 x 10 10 a I I 3.5 x 10 5.6 x 10 5.5 x 10 8 b 8 b 8 b 5.1 x 10 8 b 3.8 x 10 8 b 2.8 x 10 8 b II II 5.1 x 10 3.8 x 10 2.8 x 10 8 b 8 b 8 b 4.5 x 10 8 b 3.5 x 10 8 b 2.1 x 10 8 b III III 4.5 x 10 3.5 x 10 2.1 x 10 10 c 10 c 8 c 3.3 x 10 10 c 1.6 x 10 10 c 4.3 x 10 8 c IV IV 3.3 x 10 1.6 x 10 4.3 x 10 9 c 9 c 8 c 4.7 x 10 9 c 3.7 x 10 9 c 4.6 x 10 8 c V V 4.7 x 10 3.7 x 10 4.6 x 10

Changes in bacterial count in the five treatments at Changes in bacterial count in the five treatments at every 6- -hour intervals hour intervals every 6 6.00E+10 Bacterial count (CFU/ml) 5.00E+10 4.00E+10 3.00E+10 0 hr 6 hr 12 hr 2.00E+10 1.00E+10 0.00E+00 1 2 3 4 5 Treatment

Water quality parameters at 0, 6 and 12 hours of transport Duration Duration Treatment Treatment of of Parameter transport Parameter transport (hour) 1 2 3 4 5 (hour) 1 2 3 4 5 0 8.0 7.6 7.7 8.2 8.1 0 8.0 7.6 7.7 8.2 8.1 pH pH 6 7.7 7.3 7.5 7.7 7.5 6 7.7 7.3 7.5 7.7 7.5 12 12 7.7 7.7 7.8 7.8 7.7 7.7 7.7 7.7 7.6 7.6 0 25.2 25.4 25.5 25.8 25.3 0 25.2 25.4 25.5 25.8 25.3 Temperature Temperature 6 29.4 29.3 29.5 29.1 29.1 6 29.4 29.3 29.5 29.1 29.1 ( o o C) C) ( 12 12 30.4 30.4 30.2 30.2 30.3 30.3 30.3 30.3 30.7 30.7 0 18.6 18.2 18.5 18.6 18.2 0 18.6 18.2 18.5 18.6 18.2 Dissolved Dissolved oxygen oxygen 6 14.7 14.4 14.9 14.7 14.1 6 14.7 14.4 14.9 14.7 14.1 (mg/l) (mg/l) 12 15.0 14.5 14.3 15.4 15.4 12 15.0 14.5 14.3 15.4 15.4

Mortality (number and percent) and percent survival of size # 24 Nile tilapia fingerlings after 12 hours of transport Treatment Mortality (%) Treatment Mortality (%) 1 0.8 1 0.8 2 0.6 2 0.6 3 0.7 3 0.7 4 0.7 4 0.7 5 0.4 5 0.4

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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