RESULTS

Result of chemical and gaseous parameter changes

with probiotic (Protexin Aquatech Program)

application :

The Protexin Aquatech Program has been introduced in

which four different beneficial microbial products

containing different concentrations are taken in

combinations. Aqua media is used as a specifically

designed nutrient for the activation and

multiplication of the Bacillus spores.

4.1 Probioitic or beneficial bacterial population

reduces the ammonia concentration in shrimp

culture:

The study undertaken revealed that the Probiotic

application at the rate of 15 ml with 40 gm media

(T3) reduced ammonia concentration significantly

(P<0.01) in water in comparison with the control

pond T4. This is exhibited in the result of T3 pond

where the ammonia level was 0.0475 mg/liter which is

58

comparatively better than that of traditional

farming system as shown in control pond

T4(0.07mg/l).

The protexin treated other pond T1 where Protexin

was applied as 30 gm media + 10 ml Protexin Aqua

following the rotation technique and found that

ammonia level was reduced to 0.056 mg/l and

treatment pond T2 where Protexin application

was 35 gm Media +13 ml Protexin Aqua Program the

reduced ammonia level was 0.061 mg / litre.

Table 1 : Influence of Protexin on the ammonia content in pond water

TreatmentGiven in ponds

Ammonia concentration

MeanNH4

Standard Deviation

T1 0.0560bc

0.0587 0.097

T2 0.0612ab

0.06 0.091

T3 **0.0475c

0.046 0.09

T4 0.0702 a

0.071 0.093

59

Normal level <0.05 - -Max 0.08 -Coefficient of variation

9.93 %

Significancelevel

1 %

(T1 = 30gm media + 10 ml protexin , T2 = (35 gm media + 13 ml protexin),T3 = T3(40 gm media + 15 ml protexin), T4(without protexin)

Values are mean of 4 replications and data were analyzed after

appropriate transformations. In H2S column, means followed by same

latter(s) are statistically identical by DMRT at 1% (*) and 1% (**)

level of significance.)

4.2 Probioitic or beneficial bacterial population reduces the H2S concentration in shrimp culture:In respect of H2S content in pond water, it is

indicated that T2 treatment obviously presented the

lowest level, followed by T1 & T3. In Table 2 it

demonstrates the H2S content in pond water. It is

clearly evidenced that post pond water Probiotic

application in the T2 showed the most desirable and

significant (P<0.01) result in relation to H2S

content. Although the hydrogen sulphide content in

T3 was slightly high but this is negligible in

comparison to other probiotic treated ponds (T1 &

T2); as the ammonia content of T1 & T2 were higher

60

than T3. It is interesting to note that H2S

content in control pond T4 remain in peak most

concentration.

Table 2 : Influence of Protexin on the H2S content in pond water

TreatmentGiven in ponds

Hydrogen sulfideconcentration

MeanH2S

Standard Deviation

T1 0.015 b 0.029 0.029

T2 *0.012 b 0.021 0.02

T3 0.017 b 0.015 0.02

T4 0.027 a 0.016 0.015

Normal level <0.02 - -

Max 0.04 -

Coefficient of variation

29.44 %

Significance level

1%

61

(T1 = 30gm media + 10 ml protexin , T2 = (35 gm media + 13 ml protexin), T3 = T3(40 gm media + 15 ml protexin), T4(without protexin)

Values are mean of 4 replications and data were analyzed

after appropriate transformations. In H2S column, means followed

by same latter(s) are statistically identical by DMRT at 1% (*)

and 1% (**) level of significance.)

Although the hydrogen sulphide content in T3

was slightly high but this is negligible in

comparison to other probiotic treated ponds (T1 &

T2). It is interesting to note that Ammonia and

H2S content in control pond remain in peak most

concentration.

4.3 Probioitic or beneficial bacterial population to maintain the DO in shrimp culture:Table 3: Estimation of Dissolve Oxygen level in all probiotic(Protexin) treated pond.

62

Treatment

DO concentration mg/l

Mean Standard Deviation

DO

T1 7.50a

7.125 1.258

T2 7.00a

6.99 1.24

T3 **8.50b

8.48 1.21

T4 5.50a

5.56 1.24

63

Normal Level

- 7.0-9.0 -

Max - 9.5 -Min - 5.00 -Coefficient of variation

15.19%

Significance level

1 %

(T1 = 30gm media + 10 ml protexin , T2 = (35 gm media + 13 mlprotexin), T3 = T3(40 gm media + 15 ml protexin), T4(withoutprotexin)

Values are mean of 4 replications and data were analyzed after

appropriate transformations. In H2S column, means followed by same

latter(s) are statistically identical by DMRT at 1% (*) and 1%

(**) level of significance.)

The data as obtained are presented in Table 3 which evidenced that the DO concentration after Probioticapplication in T3 pond was found significantly higher (P<0.01) which indicate that the BOD had reduced appreciably. The result has stimulated to promote recognition of increased level of Oxygen concentration generated in pond water. The other ponds held the required marginal oxygen concentration which was 7 mg/l.(T1 : 7.50 mg/l and T2 : 7.00 mg/litre).

64

4.4 Probioitic or beneficial bacterial population reduces and PH in shrimp culture:Table 4: Estimation of Dissolve Oxygen level in all probiotic(Protexin) treated pond.

Treatment PH Mean Standard DeviationPH

T1 8.625 ab 8.525 0.751T2 8.95 a 8.95 0.73T3 *7.625 b 7.66 0.035T4 8.90 a 8.87 0.744Normal Level 7.5-8.5 - -Max 9.00 - -Min 7.5 - -Coefficient of variation

3.02 %

Significancelevel

1%

T1 = 30gm media + 10 ml protexin , T2 = (35 gm media + 13 mlprotexin), T3 = T3(40 gm media + 15 ml protexin), T4(withoutprotexin)

Values are mean of 4 replications and data were analyzed after appropriate transformations. In H2S column, means followed by samelatter(s) are statistically identical by DMRT at 1% (*) and 1% (**) level of significance.

For growth of Paenus monodon the ideal PH is from 7.5 to 8.5 , which indicated that T3 pond served to possess most suitable (P<0.05) water quality environment for shrimp culture. Although Protexin treated pond T1,T2 and control pond with out Protexin T4 were statistically identical with littledifference in T1 ; the dose recommendation of Protexin Aquatech Program in T3 showed the desired effect.

65

4.5 Effect of Probiotic to maintain Salinity Table 4: Estimation of salinity level in all probiotic(Protexin) treated pond.

Treatment Salinity Concn

Mean Standard deviation

T1 *18.00a 15.875 2.604T2 17.00a 16.12 2.514T3 16.00ab 15.89 2.221T4 12.50 12.14 2.134Normal Level 12-40 -Max 45 -Min 10 -Coefficient of variation

10.34

Significancelevel

1%

Salinity is itself a natural phenomenon in shrimp

culture pond water. In the all three Probiotic

treatment ponds T1, T2, and T3, there were increase

66

in salinity in comparison to control pond

T4 .Specially the occurrence of a

significantly(P<0.05) higher salinity was found in

T1, followed by T2 & T3. The Protexin has an optimum

effect on the bioavailability of water salinity as

found in all treatment ponds.

4.5 Effect of Probiotic to increase weight gain Table 4: Estimation of weight gain in all probiotic(Protexin) treated pond and control pond.

Treatment Weight gain(Gram)

Mean Standard deviation

T1 *31a 30.125 3.052T2 *31a 31.00 3.022T3 *33a 32.9 3.011T4 25b 25.89 3.051Normal Level 30-35 -

67

Max 45 -Min 10 -Coefficient of variation

3.54%

Significancelevel

1%

It is obvious from the above study that application

of probiotic(Protexin Aquatech) directly

interrelated with shrimp’s live weight gain. In all

probiotic(Protexin Aquatech Program) treatment ponds

the weight gain of shrimp were 31g, 31g and 33g

in T1, T2 and T3 respectively. It is noteworthy that

the body weight gain was found considerably

(P<0.01) maximum in shrimps of T3 pond, while the

shrimp of control pond were found to weigh 8 gm less

than shrimps of T3 pond. This means that shrimps

grown in probiotic(Protein Aquatech) treated ponds

exhibited appreciable higher weight gain.

68

Influence of probiotic interrelated with shrimp weoght gain potential weight gain potential and reduced ammonia content in pondwater :

Ammonia was found to have reduced up to a

desirable level in all protexin treated ponds T1,T2 and

T3 except the control pond only. The coefficient of

variance in ammonia content was found up to 9.93%, while

the correlation with weight gain was significant

(p<0.01) as revealed in experiment.

69

70

Ammoni

a

concen

tratio

n

weight gain

Graph 1: Interrelatedness between ammonia reduction

and weight gain

In relation to ammonia reduction intensity, the

weight gain was reported up to 35 gm in trial pond T3.

The data as obtained showed that the shrimp growth

promotion in T3 pond was best and this ocurred at the

ammonia concentration ranging from 0.04 to 0.05. Effect

on shrimp yield loss was found in trial pond T4 where

25 gram weight gain was evidenced in shrimp, analogously

ammonia concentration in which the pond water ranged

from 0.06 to 0.08 mg/l. The effect of high ammonia

concentration on low shrimp yield is clearly evident

in trial pond T4 and the result is presented in the graph

no 1, which shows a significant negative correlation (r

= -.767) on the rate of weight reduction relating to

actual shrimp yield. It is noteworthy that

probiotic(Protexin) application reduced the the ammonia,

71

which consequently increased shrimp yield. However the

increased yield in the treated plot was found to be due

to the combined effect of the reduction of other toxic

gas like H2S as will be indicated later. therefore the

effect of this particular reduction of ammonia on

shrimp yield enhancement could not be singled out.

Though in terms of correlation between ammonia and

weight gain , Correlation is significant at the 0.01

level (2-tailed). The other correlation result between

H2S and weight gain is quoted below.

Influence of probiotic(Protexin) interrelated

with shrimp weight gain potential and reduced

H2S content in pond water :

The record of H2S content of water was observed

throughout the entire crop cycle of shrimp culture. It

was demonstrated that the rate of H2S content varied at

different level points showing negative correlation (r =

-.767) with shrimp weight gain . As shown in graph

No 2 below when H2S exists between 0.01 to

72

0.03 mg/l the highest shrimp yield was

obtained in trial pond T3 and this was

significant at 0.01 level (2-tailed).

Graph 2 : Interrelation between H2S reduction of

pond water and shrimp weight gain

At the begening of the study the H2S concentration

of water was 0.03mg/l. During the experimental period

when H2S dropped to 0.02 mg/l in trial pond T3 , the

occurrence of weight gain in shrimp took place. Likw T3

treatment probiotic(Protexin) also influenced other

73

H2S content measurement

Weight

gain

potent

ial

treatments T1 & T2 significantly. On the contrary in

control pond T4 where shrimp weight gain was

appreciably low (25 g), showed that H2S content of pond

water remained in the rang from 0.03 to 0.08. This

level like NH4 has certainly played a role in the

shrimp weight gain and health.

Influence of probiotic(Protexin) interrelatedwith shrimp weight gain potential and increased oxygen content in pond water :

Although primarily probiotic bacteria lactobacilli

utilize DO from pond water for their own metabolism, but

it was observed afterwards that starting from the

stocking to crop maturity stage, DO oxygen increased as

probiotic was applied in pond water. The DO

concentration recorded in all treatment ponds also

varied at different time within the wide range, the

highest being at the T3 pond. As shown in graph no 3

74

that while DO reached at 8.00 mg/l, the weight gain in

T3 attained highest level.

Graph no. 3 : Interrelation between DO enhancement in pond water and weight gain

The Dissolve Oxygen resulted that there was

significant increase of dissolve oxygen in

probiotic(Protexin) treated pond water as

evidenced by reduction of BOD. The concentration of

of DO and weight gain shows a significant positive

correlation (r = -.682) . The probiotic application

is directly proportional to increase the DO, which

consequently increase shrimp yield.

Result of organoleptic traits of dressed shrimp:

75

Dissolve oxygen

Weight

gain

potent

ial

Organileptic evaluation consist in finding out

the attributes of food, in this special case of

shrimp flesh of dressed shrimp that can be

perceived by sense organs. The attributes, which

were evaluated are color, flavor, juiciness,

tenderness, texture and overall acceptability.

The six organoleptic quality traits were judged

by panel of experts and scores were given. For

each trait, highest score was 10 marks(highly

acceptable0 and lowest score was 1 mark(titally

unacceptable or spoiled). Total panel score

marks was 60. The results of test panel scores at

10 days, 20 days and 30 days were shown in tables

No X, Y and Z and in figures a to g. It is

clearly evident from tables X, Y and Z that T3 (40

gm Media +(15 ml Protexin Aqua) secured the

highest score in color, flavour, juiciness,

tenderness, texture and overall acceptability. T3

shrimp flesh secured 86.00% marks at 10 days,

76.66 % at 20 days and 71.00 marks at 30 days of

76

storage. These scores were the highest of all

other scores. On the other hand T4(with out

protexin) shrimp flesh secured 68.66 % marks at

10 days, 61.33 % marks at 20 days and 53.00 marks

at 30 days of frozen storage. These scores were

the lowest of all other scores obtained by given

different treatments.

The color attributes of the treatment pond

where 40 gm Media and 15 ml Protexin Aqua were

used (T3) obtained the highest values in

comparison to all other treatments and was found

significantly the best(P<0.01). No significant

difference was estimated among treatments other

than T3 . However, significant difference(P<0.01)

was observed when compared with no use of

Protexin Aquatech Program T4.The flavor

attributes of T3 significantly better (P<0.05)

than treatment of no use of Protexin Aquatech

Program(T4), treatment of 35 gm Media +13 ml

77

Protexin Aquatech Program(T2) and the treatment of

30 gm media + 10 ml Protexin Aquatech

Program(T1). In relation of color and flavour

there is no significant difference (P<0.05) among

T1,T2, T3 and T4. It is therefore evident from

the data that flavour score does not correspond

with color index.

The tenderness attribute of shrimp flesh from T3

(40 gm Media +15 ml Protexin Aquatech Program)

shoed significantly the best(P<0.01) of all other

treatments. Similar to tenderness attributes the

sensory scores obtained validates in favor of

juiciness, texture and overall acceptability. It

is thus concluded that the overall acceptability

of dressed shrimp flesh in respect to

organoleptic quality is very interrelated to the

sensory attributes perceived by panelists.

78

Table X: Sensory scores(color and flavour) of

microoven cooked shrimp evaluated by panel of

experts(scoring based on headonic scale:

Excellent 9-10, Very Good 7-8, Good 5-6,

Marginally accepted 3-4, Usually not

acceptable 2, Totally unacceptable or

Spoiled 0-1):

Treatment

s

Media +

Protexin

Aquatech

Program

Color Flavor

Storage(S) Mean Storage(S) Mean

10

day

s

20

day

s

30

days

10

days

20

days

30

days

T1

30 g +

10 ml

7.4 7.0 6.0 6.80b 7.4 7.0 6.0 6.74

b

T2 7.4 7.2 6.2 6.93b 7.4 7.0 6.0 6.73

79

35 g +13

ml

b

T3 = 40 g

M +15 ml

PAP

8.4 7.8 6.8 7.66a 8.4 7.6 6.8 7.60

a

T4

No protexin

7.0 6.8 5.8 6.53b

c

7.0 6.8 5.8 6.47

bc

Mean 7.5

5a

7.2

0b

6.20

c

6.98 7.55

a

7.10b 6.15c 6.47

Marks parameter: Excellent 9-10, Very Good 7-8, Good 5-6, Marginally accepted 3-4, Usuallynot acceptable 2, Spoiled 0-1Total Marks parameter: 0-20 = Unacceptable,21-30 = Acceptable, 31-40 = Good quality, 41-50 = Best quality

Table Y: Sensory scores(juiciness and tenderness)

of micro oven cooked shrimp evaluated by panel of

experts(scoring based on hedonic scale:

Excellent 9-10, Very Good 7-8, Good 5-6,

Marginally accepted 3-4, Usually not

80

acceptable 2, Totally unacceptable or

Spoiled 0-1):

Treatment

s

Media +

Protexin

Aquatech

Program

Juiciness Tenderness

Storage(S) Mean Storage(S) Mean

10

days

20

days

30

days

10

days

20

days

30

days

T1

30 g +

10 ml

7.4 7.2 6.0 7.00b

c

7.6 7.2 5.6 6.80

bc

T2

35 g +13

ml

7.8 7.4 6.4 7.20b 7.8 7.4 6.4 7.20

b

T3 = 40 g

M +15 ml

PAP

8.8 8.2 7.4 8.13a 8.8 8.0 7.2 8.00

a

T4

No protexin

7.2 7.0 5.8 6.66b

c

7.4 7.0 5.6 6.67

bc

Mean 7.80a 7.45a 6.40b 7.24 7.90 7.40a 6.20b 7.16

81

a

Marks parameter: Excellent 9-10, Very Good 7-8, Good 5-6, Marginally accepted 3-4, Usuallynot acceptable 2, Spoiled 0-1Total Marks parameter: 0-20 = Unacceptable,21-30 = Acceptable, 31-40 = Good quality, 41-50 = Best quality

Table Z: Sensory scores(texture and overall

acceptability) of micro oven cooked shrimp

evaluated by panel of experts(scoring based on

hedonic scale: Excellent 9-10, Very Good 7-8,

Good 5-6, Marginally accepted 3-4, Usually

not acceptable 2, Totally unacceptable or

Spoiled 0-1):

Treatmen

ts

Texture Overall acceptability

Storage(S) Mean Storage(S) Mean

82

Media +

Protexin

Aquatech

Program

10

days

20

days

30

days

10

days

20

days

30

days

T1

30 g +

10 ml

7.4 7.2 5.8 6.73b

c

7.4 7.2 5.6 6.73

bc

T2

35 g +13

ml

7.8 7.4 6.4 7.20b 7.8 7.4 6.4 7.20

b

T3 = 40

g M +15 ml

PAP

8.6 8.0 7.2 7.93a 8.6 8.0 7.2 7.93

a

T4

No protexin

7.0 7.0 5.6 6.53b

c

7.0 7.0 5.8 6.60

bc

Mean 7.70a 7.40a 6.25b 7.09 7.70

a

7.40a 6.25b 7.11

Marks parameter: Excellent 9-10, Very Good 7-8, Good 5-6, Marginally accepted 3-4, Usuallynot acceptable 2, Spoiled 0-1

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Total Marks parameter: 0-20 = Unacceptable,21-30 = Acceptable, 31-40 = Good quality, 41-50 = Best quality

The result of Organoleptic Quality examination

of harvested raw shrimp of Trial pond T1 which

was conducted by questionnaire among the three

groups such as Group A = high school/

college/university goers, Group B = service

holders and Group C = retired persons it was

revealed that

In trial pond T1 (Where probiotic is applied @

Protexin 10 ml + 30 gm media) was acceptable in

terms of grading standard enumerated in

methodology.

The result of Organoleptic Quality examination

of cooked shrimp of Trial pond T1 which was

conducted by questionnaire among the three groups

such as Group A = high school/ college/university

goers, Group B = service holders and Group C =

retired persons it was revealed that In trial

84

pond T1 (Where probiotic is applied @ Protexin 10

ml + 30 gm media) was good quality in terms of

taste and savor grading standard enumerated in

methodology.

85