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A Study of Ultra-Violet Irradiation on Epithelial Tissue of Fresh Water Fish, “Puntius Sophore”

Volume 32, Number 4

Vivek Sharma* and  Abhishek Srivastava

Department of Chemistry GLA University, Mathura, India.

Corresponding Author E-mail: sharmavivekgla1@gmail.com

DOI : http://dx.doi.org/10.13005/ojc/320447

Article Publishing History
Article Received on :
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Article Published : 06 Aug 2016
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ABSTRACT:

The small indigenous fishes (SIFs) are known to be micronutrient rich. The gene Puntius comprises of about 134 beautiful species, out of which PuntiusSophore is an important species. Comprehensive nutrient profile of PuntiusSophore showed that it is rich in protein and minerals. In the present study we are investigating the effect of UV irradiation on epithelial tissue of fish, PuntiusSophore. The quantities of various mineral constituents, fat and amino acids were analyzed at different time intervals. This study was undertaken to evaluate the mineral and organic chemical constituents present in scale of fish, PuntiusSophore.  The minerals (Na, Ca, Mg, P, and F), CO2 and organic constituents (fat, protein and nitrogen) were determined before and after the time intervals of 1 hour, 10 hours and 20 hours of U.V. irradiation. All mineral constituents remain unchanged after U.V. irradiation. Total nitrogen, crude fat and crude protein showed continuous decrease with increase in the time of exposure. Percentage decrease in total nitrogen, crude fat and crude protein after 20 hour exposure was 2.784, 7.79 and 2.88 respectively. Likewise Amino acids content in fish scale decreases, maximum being in case of valine after 20 hours exposure. The reason for this decrease is the C-C bond cleavage with the formation of free radicals and evolution of ammonia. Exposure also reduced the weight of the scale powder.

KEYWORDS:

U.V. Irradiation; epithelial tissue; amino acid; crude fat; crude protein; fish Puntius Sophore

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Sharma V, Srivastava A. A Study of Ultra-Violet Irradiation on Epithelial Tissue of Fresh Water Fish, “Puntiussophore”. Orient J Chem 20016;32(4).

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Sharma V, Srivastava A. A Study of Ultra-Violet Irradiation on Epithelial Tissue of Fresh Water Fish, “Puntiussophore”. Orient J Chem 20016;32(4). Available from: http://www.orientjchem.org/?p=20087

Introduction

Puntius Sophore is one of the nutritionally superior SIFs [11, 16 – 18]. Nutritional composition of fish varies with the variables as zoogeography, size, season etc. The present study was undertaken to investigate the effect of U.V. radiation on essential mineral constituents, fat and amino acids present in scales of Puntius Sophore. Ultraviolet Radiations may be understood as radiation beyond the violet region. Wavelength range of this region is 4000 to 20A0. Radiations near ultra-violet region corresponding to 2000 Å (frequency 1.51X1015 cycles / seconds) have energy equal to 1.43X1015 calories and U.V. frequency 1.5X1015 cycles/ seconds have energy 1.43X107 calories (Bajpai& Mishra, 1990) [5], (Gurdeepraj, 1991)[12] William Kemp (1986) [15]. Ultraviolet light has three wave length designations UV-A UV-B and UV-C. It has been proved that U.V. light is healthiest when it has trace amount of ultraviolet radiation. UV-B plays a vital role in the production of vitamin D3 in our skin and is essential for the absorption of calcium into bones [4]. Tuberculosis and skin lesions, too, are cured by U.V. radiation [21]. In fact before the invention of penicillin in 1938, varieties of infectious disease were cured through exposure to the sun. The ultraviolet light was found very effective in stimulating the patient’s immune system [20]. The amino acids are very important to life.  In fact some of them are absolutely essential. The results of nutritional experiments on laboratory animals show that certain amino  acids must be present in the diet, to obtain the normal growth of young and to maintain the natural state of health in adults. Some amino acids are synthesized in the animal body. The omission of any of essential amino acids from the diet leads to one or more manifestations of malnutrition. Argine, for example, is synthesized in the organism, but the rate of its formation is insufficient to permit normal growth. Some quantity of it must be provided in the food supply. [Ray Q. Brewster & William E. Mcewen (1968)] [23].

Material and Methods

A good quality Puntius Sophore fish were provided by Al-Shah enterprises Delhi.  Scales from the body of fish, Puntius Sophore were pulled out with the help of forceps. To obviate the effect of habitat and environment on the result, samples for the study were obtained from the same place during the same period of the year i.e. the first fortnight of January. Scales of fish were washed thoroughly several times with distilled water and air dried at room temp for 36 hours. Whole scales of fish were ground separately in an electrical grinder.
Moisture and Ash
Moisture was determined by keeping 1 gram of the air dried material in an aluminum cup in an electric oven at 1100C for 24 hours and then weighing the oven dried material. Ash content was determined by keeping 1gram of the material in a muffle furnace at 900oC till the weight of the ash become constant.
Minerals 
About 100 mg of ash was dissolved in the minimum quantity of HCl, calcium was precipitated as calcium oxalate. Calcium was determined volumetrically using standard KMno4 solution after liberating free oxalic acid by dissolving the precipitate in dilute H2SO4 . Fluorine was determined by the method reported by Snell & Snell, 1967 [26]. Mg was determined calorimetrically after removing calcium as calcium sulphate precipitate by taking about 500 mg of ash (Snell and Snell, 1967)26. Na was determined as sodium zinc uranyl acetate precipitate.  The colour of precipitate was intensified with a little H2O2. Sodium was estimated in Eel’s colorimeter using the green filter (Snell and Snell, 1967)26. Phosphorus was determined by dissolving 20 mg of ash in minimum quantity of HNO3. Phosphorus in canary yellow ammonium phosphomolybdate was determined by alkali metric method (Cumming and Kay, 1956) [7]. CO2 was determined with the help of Schrotter’s apparatus (Cumming and Kay, 1956 revised)7.
Fat, Protein and Amino Acids
Crude fat was extracted in a soxhlet extractor using petroleum ether (40-600C), time of extraction being 6-8 hrs7. Total nitrogen was determined by Kjeldhal method2. Crude protein was determined by multiplying the total N content by 6.25 and amino acids were determined using high performance liquid chromatography [1, 10].

Results and Discussion

Minerals are required for the normal life processes, and all animals including fish, need these inorganic elements. Fish may derive these minerals from the diet and also from ambient water. Although the minerals are required in very trace amounts but, they are vital for maintaining proper homeostasis inside the body. Puntius Sophore has significantly higher level of calcium concentration [16]which if quite high in comparison to Indian major carps (L. rohita, C. catla, C mrigala) [24]. SIFs are the rich source of calcium and other micronutrients as they are eaten whole with bone head and eye [11]. The bioavailability of calcium in Puntius Sophore is as high as that from milk [13]. Deficiency of calcium may be associated with rickets and osteomalacia. Na and F are present in traces, Mg content is too low and ash accounted for 0.2065 (Table. 1&2), calcium and phosphorus are the major constituents of the fish scale. CO2 content is also low as compared to calcium and phosphorus oxides (Table. 1). After 1 hour U.V. irradiation moisture content of air dried scale decreases extensively while the minerals (Na, Ca, P, Mg, and F) and CO2 remain unchanged. After 20 hours of U.V. exposure appreciable decrease in moisture content is observed in air dried scales but minerals (Na, Ca, P, Mg and F) and CO2 still remain unchanged (Table. 2).
Table 1: Mineral Composition in Epithelial Tissue (Scales) of Fish Puntius Sophore Before Irradiation (Gm/100gm Of The Dry Matter)

 

Average of 10 Fish

 

Moisture

Ash

Mineral

Values

Mineral

Oxide / Fluoride

Values

Ash (unaccounted)

 

Length

(cm)

Girth

(cm)

Weight

(gm)

8.66

8.52

89.9

10.74

27.61

Ca

Mg

Na

P

F

10.49

0.33

0.0026

4.68

0.0016

CaO

MgO

Na2O

P2O5

CaF2

CO2

      14.6

0.55

0.0035

12.25

0.2065

Total=

27.4035

0.003

1.003

Table 2: Mineral composition in Epithelial Tissue (Scales) of Fish Puntius Sophore after irradiation (gm/100 gm of the dry matter)

Time of irradiation

Moisture

Ash

Minerals

 

Mineral

oxides

Ash (unaccounted)

1 HOUR

10.01

27.61

Ca

Mg

Na

P

F

10.49

0.33

0.0026

4.68

0.0016

CaO

MgO

Na2O

P2O5

              CaF2

CO2

14.6

0.55

0.0035

12.25

0.2065

TOTAL =27.4035

0.0033

1.003

10 HOURS

9.30

27.61

Ca

Mg

Na

P

F

10.49

0.33

0.0026

4.68

0.0016

CaO

MgO

Na2O

P2O5

CaF2

CO2

14.6

0.55

0.0035

12.25

0.2065

TOTAL=27.4035

0.0033

1.0003

20 HOURS

9.01

27.61

Ca

Mg

Na

P

F

10.49

0.33

0.0026

4.68

0.0016

CaO

MgO

Na2O

P2O5

CaF2

CO2

14.6

0.55

0.0035

12.25

0.2065

TOTAL=27.4035

0.0033

1.003

The fish is a rich source of fat soluble vitamins like A, D, E and K. Puntius Sophore  contains saturated fatty acids (20.02%), monounsaturated fatty acids (37.12) and high amount of polyunsaturated fatty acids (PUFAs)[16]. PUFAs have potential in reducing coronary heart disease, atherosclerosis, cancer, arthritis, hypertension and Alzheimer [3, 8, 14, 22, 25, 29, 30,]. The percentage decrease in fat content after 1, 10 and 20 hours of exposure are 2.72, 5.51 and 7.79 respectively. This decrease is observed due to degradation of fat through free radical formation (Table. 4).

Table 3: Crude Fat, Nitrogen and Crude Protein Contents of Fish Puntius Sophore before irradiation (gm/100gm of dry matter)   

Average of 10 fish

Crude Fat

Total Nitrogen

Crude Protein

Length (cm)

Girth (cm)

Weight (gm)

8.66

3.91

89.9

0.308

11.53

72.08

Total of Ash + Crude Protein + Crude Fat= 99.998

Table 4: Crude fat, Total Nitrogen and Crude protein content in scales of Fish Puntius Sophore after U.V. irradiation (gm/100 gm of the dry matter)

Time of irradiation

Crude Fat

% decrease of crude fat

Total Nitrogen

Decrease of nitrogen

After irradiation

Crude protein

(N×6.25)

Crude protein

(after irradiation)

% decrease of crude protein

1 hour

0.301

2.27

11.474

0.520

72.08

71.68

0.554

10 hours

0.291

5.51

11.294

2.047

72.8

70.56

2.108

20 hours

0.284

7.79

11.209

2.784

72.8

70.00

2.88

Total (Ash+ Crude fat + Crude protein) = 99.991 (After 1 hour of exposure)

Total (Ash+ Crude fat + Crude protein) = 99.98 (After 10 hour of exposure)

Total (Ash+ Crude fat + Crude protein) = 99.97 (After 20 hour of exposure)

Proteins are essential to all life. In animals they help from supporting and protective structures such as cartilage, skin, nail, hairs. They are major constituents of enzymes, antibodies, hormones and body fluids. Essential amino acids meet the demand of growth during childhood. Fish protein contains essential amino acids that are required for the human nutrition and thus improve the overall protein quality of a diet [19]. The predominant amino acids present in Puntius Sophore are histidine followed by glutamic acid, aspartic acid, serine, threonine and lucine[16].   Several reports are available to check the effect of U.V radiation on the fish collagen; the U.V radiation causes the progressive degradation of the collagen molecules into smaller molecular fragments [6, 27]. Collagen peptide is beneficial as a dietary supplement to suppress UV-B-induced skin damage and photo aging [28].Continuous decrease in protein content is observed after 1 (0.554), 10 (2.108) and 20 (2.88) hours of radiation (Table. 3, 4). Amount of different amino acids in fish scales have also been determined after 1, 10 and 20 hours of U.V. radiation, after one hour of exposure maximum decrease is observed for glutamic acid (1.704%) while cystine shows minimum decrease (0.369%), after 10 hours of exposure the maximum percentage of phenylalanine (2.821) decreases maximum and percentage of lysine (1.754) decreases minimum, valine decreases maximum (4.736%) while cystine decreases minimum (3.367%) after the exposure of 20 hours of radiation (Table. 6). Table 2 clearly indicates that there is no change in mineral constituents after 1, 10 and 20 hours of radiation; the change is only in moisture content. Amino acids are decreased continuously with the time of exposure (Figure. 1). U.V. radiation was found useful to improve the gel strength of fish gelatin [9].

Table 5: Percentage of Amino Acids in scales of fish Puntius Sophore before irradiation (gm/100gm of the dry matter).

S.No

Name of amino acid

Value

1

Cystine

8.35

2

Lysine

2.41

3

Histidine

3.71

4

Arginine

4.95

5

Hydroxyproline

5.28

6

Cysteine

2.46

7

*  Serine

24.44

8

Aspartic Acid

4.91

9

Glycine

4.14

10

Glutamic Acid

3.71

11

Threonine*

6.01

12

Alanine

4.64

13

Tyrosine

1.92

14

Valine

3.85

15

Proline

4.13

16

Methionine

2.78

17

Isoleucine

3.66

18

Leucine

2.46

19

Phenyl Alanine

3.17

20

Tryptophan

3.02

Total

100.00
Table 6: Percentage of Amino Acids of Scales of Fish Puntius Sophore After Irradiation (Gm/100gm Of The Dry Matter).

Amino Acid

Values after irradiation at different time intervals

1 hour

10 hours

20 hours

 

value

% decrease

Value

% decrease

Value

% decrease

Cystine

8.319

0.369

8.147

2.431

8.069

3.367

Lysine

2.393

0.697

2.368

1.754

2.327

3.441

Histidine

3.672

1.021

3.615

2.554

3.541

4.550

Arginine

4.913

0.747

4.837

2.277

4.755

3.933

HydroxyProline

5.217

1.195

5.143

2.598

5.059

4.188

Cysteine

2.431

1.171

2.393

2.716

2.352

4.375

*Serine

24.312

0.524

23.881

2.288

23.524

3.749

Aspartic Acid

4.888

0.454

4.786

2.518

4.730

3.665

Glycine

4.103

0.903

4.048

2.221

3.971

4.077

Glutamic acid

3.647

1.704

3.615

2.554

3.541

4.550

Threonine*

5.926

1.397

5.856

2.568

5.767

4.042

Alanine

4.609

0.665

4.532

2.332

4.452

4.056

Tyrosine

1.899

1.075

1.871

2.538

1.846

3.829

Valine

3.824

0.674

3.768

2.130

3.668

4.736

Proline

4.075

1.337

4.023

2.601

3.971

3.845

Methionine

2.748

1.160

2.704

2.741

2.666

4.100

Isoleucine

3.621

1.053

3.577

2.266

3.529

3.591

Leucine

2.431

1.171

2.393

2.716

2.352

4.375

Phenyl Alanine

3.140

0.937

3.081

2.821

3.035

4.249

Tryptophan

2.988

1.049

2.953

2.209

2.884

4.518

TOTAL

99.157

97.591

96.040

  N- Terminal   residue not determined

-CONH2   not  determined

*Corrected for the loss during hydrolysis

Figure 1: Comparative study of amino acid content before and after U.V. irradiation of different time exposure Figure 1: Comparative study of amino acid content before and after U.V. irradiation of different time exposure                  

Click here to View Figure

 

Conclusion

The entire study leads to the conclusion that moisture content shows a decrease as result of U.V. irradiation. Crude protein, crude fat and total nitrogen decrease with the time of exposure. Minerals remain unchanged as a result of U.V. irradiation. The individual amino acids also decrease with the time of exposure.

 Acknowledgement

The author acknowledge the management of GLA University, Mathura for financial support and Principal K. R. (PG) College, Mathura for allowing to conduct experiment in their laboratory .The author also acknowledge the vender Al shah Enterprises for providing fish.

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