Pornthep Niamphithak, Siripavee Chareonwattanasak
and Sompong Doolgindachbaporn*

Department of Fisheries, Faculty of Agriculture, Khon Kaen University, Khon Kaen 40002, Thailand.

Abstract

This study was investigated the effect  of  Probiotic (L. plantarum) supplementation at 0 , 1×107,1×108,1×109cfug-1 diet  on growth performance, feed utilization, survival rate and carcass quality in Bocourti’s catfish(Pangasiusbocourti).An average initial weight of fish 132.59 ±1.20 gfish-1were divided into four treatments, three replication and fed for 8 weeks. The results showed that body weight, weight gain, daily growth rate, specific growth rate, survival rate, feed conversion ratio and feed conversion efficiency were not significant difference in all groups (P>0.05).However, Fish fed dietary supplementation of probitic at 1×107 cfug-1 diet  had trendy high in body weight , weight gain, daily growth rate  and specific growth rate with207.16±16.31 g., 74.30±15.13 gfish-1, 1.33±0.27gday-1 and  0.79±0.13%day-1respectively. The biochemical parameters in serum and carcass quality were did not significantly different for any of the diet treatment. Therefore, this study indicated that the optimum level of L. plantarum in diet for bocourti catfish culturing could be used for contains  1×107cfug-1diet

Keyword: Bocourti catfish (PangasiusbocourtiSauvage, 1880) growth performance.

Introduction

Bocourti catfish or Basa or Pla-mong (Thai name) is common name of PangasiusbocourtiSauvage 1880 that number is Pangasiidae. This fishes are native species in Mekong and Chao Phraya basins in Indochina. In Thailand found in Mae Kong river, Chao Phraya river and some other smaller river such as Mun river and Chi river and other river in Northeastern in Thailand23,25. This fish has a good taste and has been accepted of consumers and the demanded of globe has been increased in America and Europe15.In both foreign markets are mostly demanded in filet forms and has high price. In Thailand markets had price depended on size of fish, e.g, 0.7-1.0 kg had about up to 50 baht and 1.5 to 2.0 kg had about 150 baht21.

Recently, Aquaculture is important sector and spreading area to providing human consumption and fisheries. The growing of aquaculture practices  that  from many  factor:1) the demand of market  because protein form the aquatic animals is good quality, 2) the security and safety food  of consumer and 3) make income from aquaculture5.However, aquaculture practices in intensive system requires cultivation at high densities, which has caused destroyed the environment because the effluent water from the ponds containing  organic wastes in high volume,effect on water quality  such as reducing dissolved oxygen, giving the ammonia, nitrites  and hydrogen sulfide from the  metabolites of organics degradation that often areresponsible for mortality27.And, under these conditions of intensive system, aquatic animals has to high-stress, that easily infection by pathogens and high mortality rate and causing a decreasing productivity3.So, the owner of farm using antibiotic drug for preventing and treatingwhich has seen the previous research that can be used to prevent infection.But, the using antibiotics will kill the bacteria in the digestive tract of aquatic animals andthe accumulation of antibiotic residuesin fish products to be harmful for human consumption. Form the this problem, the farmer has turned to use probiotics in aquaculture that it is non-pathogenic microorganisms in aquatic animals that are many proportion , such asimproved growth of fish , preventing the pathogen , increasingutilization of nutrient , treat  thewater quality in pond, reduce stress of fish and improved reproductivesystem of aquatic animals5.

Probiotic is a relatively new term which is used to name microorganisms that are associated with the beneficial effects for the host. Probiotics have been introduced as a growth stimulant.Such as the study of Lara et al.17 that use of Streptococcus as growth promoters on  Nile tilapia (Oreochromisniloticus) the result showed probiotic strain,increasing significantly the content of crude protein and crude lipid in the fish, also weight has increased from 0.154 g to 6.164 g. in 9 weeks of culture. And ornamental fishesinclude swordtail (Xiphophorushelleri, X. maculatus) andguppy, (Poecilia reticulate, P. sphenops), their feed wassupplemented with Bacillus subtilisand Streptomyces, can be increases in growth and survival of Xiphophorusand Poeciliaafter 90 and 50 days of experimental period, respectively7,12.

Lactic acid bacteria (LAB) are most applied probioticsin foruse enhanced growth or immune systemfor aquaticspecies8,9,24.And they are producers of bacteriocins and organic acids (lactic and acetic acids) which have inhibitory effects in vitro on the growth of some pathogens in fish20.And  they have the previous study for use enhanced growth or immune system in fish . e.gNile tilapia,Oreochromis niloticus1,Grouper,Epinephalus coioides26, Cobia,Rachycentron canadum11 and European sea bass, Dientrachus labrax19.Therefore, The aim of this study was to determine the effect of Lactobacillus plantarumsupplementationon growth performance, survival rate, serum biochemical and carcass quality in Bocourti catfish (PangasiusbocourtiSauvage, 1880)

 

 Materials and Methods

Fish, diets and feeding protocol

This study was carried out with the used four different levels of Lactobacillus  plantarum(0,1×107, 1×108 and 1×109 cfug-1 diet ). A Completely Randomized Design (CRD) with three replication. The basal diet was developed from formula diet of Boonarsa and Doolgidachbaporn4and the nutritional value of basal diet had a crude protein and gross energy with 29.89% dry matter and 424.65 kcal per 100 g diet(Table 1).The experimental diets were prepared by mixing the dry ingredients with soybean oil and water by hands and then the wet dough was placed in a grinder and pelletedby mincer to pass through a 2-mm and finally the moist pellets were dried in hot air oven at 60 °C for 12 hrand then stored in the fridge at -20 0C for further uses.

LactobacillusplantarumCR1T5 were provided from Department of  Microbiology, Faculty of science, KhonKaenUnversity, Thailand.The bacterial suspension content was about 3.4×109 cfuml-1 and the bacterial were prepared stocking every twice a weeks. After that diluted the bacteria in concentration of experimental by used sterilized MRS  broth is dilutant. The dilution of bacteria sprayed on basal diet with 0, 1×107, 1×108 and 1×109 cfug-1 diet that test diets were daily prepared.

Bocourti catfish(P. bocourti) juvenile obtained from Phayao Inland Fisheries Research and Development Center, Phayao Province, Thailand . Fish were kept in 1000L fiber tank for acclimatization with commercial diet with 40% protein for four weeks. Then protein content in diet was decreased to 30 and fed for 8 weeks prior to feeding the experimental diets.After the acclimatization fish were randomly distributed into four groups with three replications  that  each replicate contained 10 fish (mean wet weight of 132.59±1.20gfish-1) in an aquarium ( 60 x 150 x 60 cm3) filled the water 450 L. which 12 aquaria tanks. Water quality parameters such as dissolved oxygen (6.05±0.28 mg L-1), temperature (26.98±0.19°C) and pH value (7.37±0.27) were measured every day during the experimental period.

During the trial, fish were fed ad-libitum by hand fed method twice a day (08:00 and 15:00 h) for 8 week. Feed consumption was recorded weekly and fish from each tankwere weighed to measure growth every two weeks until the end of the experiment; growth performance and feed utilization were calculated.

Evaluation of growth performance and feed utilization parameters

The collected data on weight gained  (WG), average daily gain (ADG), specific growth rate (SGR), feed conversion ratio (FCR), feed conversion efficiency (FCE) and  survival rate (SR) were calculated.

WG (g) = Final Weight –  initial weight

ADG (g/day) =( Final Weight –  initial weight)/period of experiment

SGR (% /day) = (Ln final weight – Ln initial weight)/days x100

FCR = total feed intake (g)/total wet weight gained (g)

FCE (%) = total wet weight gained (g) / total feed intake (g) x100

Survival rate (% ) =  (final fish number)/(initial fish number).x100

 

Serum biochemistry and carcass quality

At the end of the experiment, after fish were starved for 12 hours, randomized two fish per tank that were anesthetized with tricainemethanesulfonate (MS-222).And blood samples were collected from caudal vein that kept in non-heparinized tubes.Hematocrit was determined by using microhematocrit centrifuge. The serums were separated into aliquots and analyses of serum biochemistry. Serum was analyzed for total protein (TP) and alkalinephosphatase (ALP) activity. The serum biochemical indices were determined with an automatic biochemical analyser (BS-200; Mindray,Shenzhen, China). Then the fishes were killed andliver and  visceral were keeped and test investigate weighed carcass quality by Hepatic somatic index (HSI), Visceral Somatic index (VSI), Visceral Fat index ( VFI) and Muscle ratio (MR) by equation following:

HSI (%) = (Liver mass(g)/ body mass (g)) × 100

VSI (%) = (total inner organ mass (g) / body mass (g)) × 100

VFI  (%) = (Visceral Fat mass (g)/ body mass (g)) × 100

MR (%) = muscle mass (g)/ body mass (g)) × 100

Statistical analysis

The data were subjected to one-way analysis of variance (ANOVA) and if significant differences (p<0.05) were found, Duncan’s multiple range test was used to rank the groups.

 

Result

Growth performance, feed utilization and survival rate

The growth performance, feed utilization and survival rate of bocourti catfish (P.bocourti) with an initial average weight of 132.59±1.20 gfish-1 fed with diet containing Lactobacillus plantarumat 0, 1x 107, 1×108 and 1×109 cfug-1 diet for 8 weeks is shown in Table 2. At the end of experiment found that growth performance base on the weight gain (WG), average daily gain (ADG) and  specific growth rate (SGR) and feed utilization base on feed conversion ratio (FCR) and feed efficiency ratio (FCE) were not significant different (p>0.05). But fish fed with Lactobacillus plantarum at 1x 107 cfug-1 diet had maximum growth performance  and best feed utilization compared with fish in other group. The weight gain (WG), average daily gain (ADG), specific growth rate (SGR), feed conversion ratio (FCR) and feed efficiency ratio were rangedfrom  64.30±9.66 to 74.30±15.13 gfish-1. 1.15±0.17 to 1.33±0.27 dday-1, 0.70±0.09 to 0.79±0.13 %day-1, 1.91±.41 to 2.17±0.31 and 46.76±7.04 to 54.06±11.01 %, respectively. And survival rate was 100 % in all groups.

 

Table 1 Ingredient and nutritional value of basal diet

Ingredients (%crude protein) Content (gKg-1)
Fish meal (57.48) 300.00
Soybean meal (44.00) 180.20
Broken rice (6.62) 255.00
Rice bran (11.73) 11.00
Corn meal(7.22) 93.00
Full Fat Soybean(35.55) 68.16
Soybean oil 35.00
Alpha starch 50.00
Vitamin and mineral1 7.64
Proximate composition based on dry matter (%)
Crude Protein 29.89
Moisture 6.79
Fat 7.26
Fiber 2.52
Ash 7.91
Nitrogen free extract  (NFE)2 45.63
Gross Energy (GE ;Kcal/100 g)3 424.65

 

Vitamin and mineral mixture(g) :retinyl acetate ( 100 IU.g-1) 0.03 g., Cholecalciferol (20IU.g-1) 0.03 g. D,L-α-tocopherol acetate 0.009 g., menadione 0.00525 g., thiamine nitrate 0.00375 g., riboflavin 0.006 g. Pydroxinehydrochlorite, 0.006 g., D-calciumpantothenate 0.015 g., Niacin 0.015 g., Folic acid 0.0015 g. Cyanocobalamin 0.00045 g. ascorbyl acetate 0.03 g. CaCO3 6.00 g.MgSO4:7H2O 1.00 g. ZnSO4:7H2O 0.60 g. FeSO4:7H2O 0.03 g.  CuSO4:5H2O 0.06 g. and  KI 0.001 g.

2NFE = 100-(% Moisture + % Crude Protein + % Fat + % Fiber + % Ash) 18, NRC (1993)

3 Gross Energy = (%crude protein x 5.64) + (% NFE x 4.11) + (%fat x 9.44) 18; NRC (1993)

 

Table 2 Growth performance, feed utilization and survival rate of bocourti catfish
(P. bocourti) fed with experimental diets for 8 weeks

Parameter Levels of Probiotic( L. plantarum) supplement in diet (cfug-1) p-value
0 1×107 1×108 1×109
initial weight (g) 132.51±1.38 132.84±1.44 132.46±1.35 132.69±0.90 0.982
Final Weight (g) 197.86±9.56 207.16±16.31 198.25±8.64 196.99±10.35 0.690
WG (g) 65.37±8.23 74.30±15.13 65.80±8.50 64.30±9.66 0.660
ADG (gday-1) 1.17±0.15 1.33±0.27 1.17±0.15 1.15±0.17 0.657
SGR (%day-1) 0.72±0.07 0.79±0.13 0.72±0.08 0.70±0.09 0.661
FCR 2.13±0.26 1.91±0.41 2.11±0.28 2.17±0.31 0.747
FCE (%) 47.53±5.95 54.06±11.01 47.85±6.15 46.76±7.04 0.657
SR (%) 100 100 100 100

Means within row followed by a common letter are not significantly different   according to DMRT (P < 0.05)

 

Table 3 Carcass quality of bocourti catfish (P.bocourti) fed with experimental diets for 8 weeks

Parameter Levels of Probiotic( L. plantarum) supplement in diet (cfug-1) p-value
0 1×107 1×108 1×109
HSI(%) 1.81±0.20 1.63±0.35 1.78±0.30 2.08±0.34 0.393
VSI(%) 9.92±3.33 9.17±1.08 9.51±1.47 10.25±0.68 0.908
VFI(%) 3.50±1.24 2.96±1.41 3.67±1.63 3.02±0.64 0.879
MR(%) 34.97±1.70 33.81±3.07 34.69±1.66 32.68±0.82 0.522

Means within row followed by a common letter are not significantly different   according to DMRT (P < 0.05)

 

Table 4.Haematocrit and serum biochemistry ofbocourti catfish (P.bocourti) fed with experimental diets for 8 weeks

Parameter Levels of Probiotic( L. plantarum) supplement in diet (cfug-1) p-value
0 1×107 1×108 1×109
Hct(%) 32.25±0.50a 34.25±1.26ab 36.75±1.85b 35.13±0.25b 0.001
TP (gd-1)l 4.107±0.931 3.390±0.577 3.527±0.532 3.673±0.278 0.556
ALP (Ul-1) 51.000±29.597 61.333±50.402 82.667±31.943 91.000±38.158 0.579

Means within row followed by a common letter are not significantly different   according to DMRT (P < 0.05)

Carcass quality

The termination of experiment, thecarcass quality  was determine in Bocourti catfish (P. bocourti)  fed  withtest diets containing Lactobacillus plantarum  at 0, 1x 107, 1×108 and 1×109 cfug-1(Shown in Table 3). The Hepatosomatic index (HSI), vesicle somatic index (VSI), Vesicle fat index (VFI) and muscle ratio (MR)were not significant different (p > 0.05).  Hepatosomatic index (HSI), Visceral somatic index (VSI), Visceral fat index (VFI) and Muscle ratio (MR) were ranged from 1.63±0.35to 2.08±0.34%, 9.17±1.08 to 10.25±0.68%,2.96±1.41 to 3.67±1.63 % and 32.68±0.82to 34.97±1.70 %, respectively

Serum biochemistry

The hematocrite(Hct) of fish fed with diet containing at L.plantarum  at 0, 1x 107, 1×108 and 1×109 cfug-1were determined at the end of the experiment that was highly significant different (p<0.01).and shown in Table 4. The hematocrit of fish fed with diet supplementary L.plantarum  at 1×108 and 1×109 cfug-1diet differ control group ( 0 cfug-1diet) but did not differ with 1x 107 cfug-1diet. And there were 32.25±0.50, 34.25±1.26, 36.75±1.85 and 35.13±0.25 % of fish fed with diet containing at L. plantarum  at 0, 1x 107, 1×108 and 1×109 cfug-1.respectively. The fish serum from all groups were analysed  total protein (TP) and alkaline phosphatase (ALP) on the end of experiment period that were not significant different (p>0.05). The total protein was ranging from 3.390±0.577 to 4.107±0.931 gd-1 and alkaline phosphatase (ALP) was ranging from 51.000±29.597 to 91.000±38.158 Ul-1. (Table 4)

Discussion

This study wasevaluated the effect of using Lactobacillus plantarum levels on Growth performance, Survival rate, Serum biochemical and Carcass quality of bocourti catfish (PangasiusbocourtiSauvage, 1880). From the present study, the growth performance and feed utilizationsof fish were not significantly different between groups. But, the growth and feed utilization of fish fed with L. plantarumat1x107 cfug-1 diet was better than other groups. Therefore, it can be concluded that giving optimal level of probiotics improved growth of fish which was consistent with study of Piccolo et al.19 that use L.plantarum in diet of European sea bass (Dientrachuslabrax) and no did not significantly different between groups. However the additions L.plantarum to dietincreased growth in term of weight gain, specific growth rate and protein efficiency ratio were 3.6, 5.2 and 6.9 % respectively.Lara et al.17reported that use of Streptococcussp.as growth promoters on  Nile tilapia (Oreochromisniloticus) increasing significantly the content of crude protein and crude lipid in the fish, also weight has increased from 0.154 g to 6.164 g. in 9 weeks of culture.Similar resulted were conducted in swordtail (Xiphophorushelleri, X. maculatus) and guppy, (Poecilia reticulate, P. sphenops), their feed was supplemented with Bacillus subtilisand Streptomyces sp. can be improved in growth and survival of swordtail andguppyafter 90 and 50 days of experimental period. , respectively7,12. These effects have been reported a similar response and positive effect on productive performance in other species. e.g.Alatic cod, Gadus morhua13, Juvenile Dentex, Dentexdentex L.14.

The carcass quality of fish in these study found that hepatosomatic index (HSI), vesicle somatic index (VSI) vesicle fat index (VFI) and muscle ratio (MR) were not significantly different between groups (p>0.05). Likely, the study of Dawood et al.6 that used L. rhamnosus and Lactococcuslactis supplement in diet of red sea bream(Pagrus major) at 106 cellg-1was found the hepatosomatic index (HSI) and somatic index (VSI) were not significantly different (p>0.05) which was consistent with study of Piccolo et al.19 that use Lactobacillus plantarum in diet of European sea bass (Dientrachuslabrax).

Hematocrit has been recognized as valuable tools for monitoring of fish health2,18. From the present study the hematocrit value of fish from all groups and has significantly different(p<0.01). And the trend of hematocrit increased on probictics levels in diet.The results indicated that supplement of probiotic imporvedhealthy of fish.The blood protein, albumin and globulin are the major proteins, which play a significant role in the immune response22. From this study showed not significantly different of total protein in fish serum. Because when liver is damaged, it will total protein, albumin and globulin in serum decreased16.The alkaline phosphatase (ALP) in these study was not significantly different. which was consistent with study of Piccolo et al.19 that use Lactobacillus plantarum in diet of European sea bass (Dientrachuslabrax).However, the trend of ALP increased on supplementary Lactobacillus level in diet of bocourti catfish. From the reported ofPiccolo et al.19 that highest values of ALP in fish resultedof increased growth rate and feeding level.

Conclusion

In conclusion, this present study indicated that supplementation of different  concentrations Lactobacillus plantarum in diet of Bocourti catfish (P. bocourti) were not significantly different in growth performances, feed utilizations, carcass quality and serum biochemistry. The haematocrit values was increased with increased concentration of probiotics. Base on these results, use ofLactobacillus plantarum at 1x 107 cfug-1supplementation in diet had maximum growth performance  and best feed utilization.

 

Acknowledgement

This work was supported by the Higher Education Research Promotion and National Research UniversityProject of Thailand, Office of the Higher Education Commission, through the Food and Functional Food Research Cluster of KhonKaen University

 

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