EFFICACY OF MICROBES IN BIOREMEDIATION OF TANNERY...

RESEARCH ARTICLE

EFFICACY OF MICROBES IN BIOREMEDIATION OF TANNERY EFFLUENT

1* Aneez Mohamed, M., 1 Sekar, P and 2 George John 1Department of Zoology, Jamal Mohamed College (Autonomous), Tiruchy –20 2PG Department of Zoology, Periyar E.V.R. College (Autonomous), Tiruchy -23

ARTICLE INFO ABSTRACT

The present research work has been carried out to analyze the physico-chemical characteristics of tannery effluent and to reduce the load of BOD and COD by the process of bioremediation using microorganisms. Among the physical chemical parameters analyzed TDS, TSS, BOD and COD were found to be very high. One bacterium, Bacillus cereus and one fungal species, Aspergillus niger were identified and isolated and used in the reduction of BOD and COD in the tannery effluent. Four different concentrations of untreated tannery effluents (control, 25, 50, 75 and 100%) were prepared. The experiment was carried out for 72 hrs (3 days). It is inferred from the results that Aspergillus niger found to be more effective in reducing BOD and COD than the bacterium, Bacillus sp. in the untreated tannery effluents.

INTRODUCTION Industrial revolution has generated unprecedented disturbances in the environment due to the introduction of anthropogenic pollutants such as organic, inorganic and xenobiotic chemicals in the form of untreated industrial waste water. With increasing population and industrial expansion, the need for the treatment and disposal of the waste has grown. The tannery effluent waste are ranked as high pollutants among all other industrial wastes(Eye and Lawrence 1971).Tannery industrial waste water is a serious consequence from the pollution point of view for streams, Freshwater and land used for agriculture. The lack of awareness in the modern industrial practice has resulted in the discharge of tannery effluents which exhibit very high value for Cr, Sulphide, and chloride, TDS, TSS, BOD and COD in the water stream or land. In the course of the last two decades a wide variety of technologies had been developed for cleanup operations of contaminated sites. Bioremediation has evolved as the most promising one because of its economical safety and environmental features, since organic contaminants become actually transferred and some of them are fully mineralized. Bioremediation of tannery effluents is an attractive environment friendly, safe and cost effective alternative technology to conventional methods. Microbes in the environment play an important role in cycling and destroying them through bio-degradation. The present investigation essentially deals with the characterization tannery effluents and to biodegrade certain important parameters viz. BOD and COD using microbes like fungus, Aspergillus niger and bacterium, Bacillus cereus in the untreated tannery effluents. *Corresponding author: [email protected]

MATERIAL AND METHODS

In the present study, effluent was collected from a tannary at Sembattu in Tiruchirappalli District, Tamil Nadu, India. The effluent samples were collected raw as well as from final discharge point where in effluent from all the stages of processing are released together.The effluent was collected in polythene containers [1 litres capacity] and were brought to the laboratory with due care and stored at 20oC. The physico-chemical parameters such as pH, Electrical conductivity (EC), Total dissolved solids (TDS), Total suspended solids (TSS), Biological oxygen demand (BOD), Chemical oxygen demand (COD), Oil and grease (OG) and Chloride (Cl) and sulphate of the effluent were analyzed following standard methods outlined by APHA (1989). The isolation of bacteria from the tannery effluent was done by serial dilution technique. (Charurvedi, 1992). After identification of the bateria, culture was carried out in the laboratory to be used for biotreatment of the tannery effluent. Bactrerium such as Bacillus sp. was used for the treatment of tannery effluent. Tannery effluents collected were used for the analysis of Mycoflora. The fungi were isolated using dilution plating method. The fungi were mounted in lactophenol cotton blue and observed through light microscope. They were identified using standard manuals. (Gilmann, 1967; Subramanian, 1971, Ellis, 1971; Barnett and Hunter, 1972). After identification of the fungi, culture was carried out in the laboratory to be used for biotreatment of the tannery effluent. Fungi such as Aspergillus niger, was separately used for the treatment of tannery effluent.

ISSN: 0975-833X

Available online at http://www.journalcra.com

International Journal of Current Research Vol. 33, Issue, 4, pp. 324-326, April, 2011

INTERNATIONAL JOURNAL OF CURRENT RESEARCH

Article History:

Received 27th January, 2011 Received in revised form 17th February, 2011 Accepted 19th March, 2011 Published online 27th April 2011

© Copy Right, IJCR, 2011 Academic Journals. All rights reserved.

Key Words: Microorganisms, Bacillus cereus, Aspergillus niger, Bioremediation, Tannery effluents, BOD, COD.

Bioremediation of Tannery Effluent Using Fungus and Bacterium Approximately 10 g (fresh weight) of mycelia of Aspergillus niger, and 10 ml (fresh weight) of Bacillus sp. were transferred to experimental jars containing 1000 ml of different concentrations (25%, 50%, 75% and 100%) of tannery effluent. They were kept in an orbit shaker for 72 hrs and maintained at 28 2 C. Two important physicochemical parameters such as BOD and COD were estimated before and after 72 hrs (3 days) to check the degradation process by the fungus and the bacterium.

RESULTS The water quality parameters of the tannery effluent were analyzed and its results are presented in the Table 1. The results of the analysis showed that the tannery effluent was grey coloured with a disagreeable odour, acidic pH, with high organic and inorganic load indicating high EC, BOD,COD,TSS,TDS, chloride and sulphate. Based upon the dominant fungus and bacterium, Aspergillus niger and Bacillus cereus were individually used to reduce the BOD and COD in the effluent. Laboratory scale biodegradation of

Table 1. Water quality parameters of Tannery effluent at two different units

Parameters Inlet. (Raw) Outlet – Final

(Treated)pH Electrical Conductivity (dsm-1) TSS (mg/1) TDS (mg/1) Chloride Sulphate BODCOD Oil & Grease

10.4 3.90 1600 2510 1600 570 947

2277 0.61

6.9 2.90 100 1480 1450 110 230520 0.02

Table 2. BOD of tannery of effluent before (control) and after biodegradation (72 hrs) using fungus and bacterium

Parameters Concentration of the effluent

Mean + SD & % Reduction

Control Aspergillus niger Bacillus cereus

BOD

100% Mean ± SD 947.5 ± 42.98 904.16 ±29.73 921.7 ± 35.30 % Reduction -4.57% -0.63%

75% Mean ± SD 834.16 ± 27.28 620 ± 50.49 658.3 ±44.45 % Reduction -25.67% -21.08%

50% Mean ± SD 635.83 ± 26.72 481.33 ±23.47 528.3 ± 44.00 % Reduction -24.29% -16.92%

25% Mean ± SD 311.66 ± 58.70 252 ± 59.74 280.8 ± 26.53 % Reduction -19.14% -9.91%

Table 3. COD of tannery of effluent before (control) and after biodegradation (72 hrs) using fungi and bacteria

Parameters Concentration of the effluent

Mean + SD & % Reduction Control Aspergillus niger Bacillus cereus

COD 100% Mean ± SD 2277.83±72.47 1835.5±39.97 1938.3±32.62 % Reduction -19.42% -14.91%

75% Mean ± SD 1772 ± 42.66 1576.7±31.25 1676.6 ±29.26 % Reduction -11.02% -5.38%

50% Mean ± SD 1452.5 ± 30.12 1362±22.36 1404.16±22.22 % Reduction -6.64% -3.33%

25% Mean ± SD 1355 ± 64.18 1117.5± 33.27 1216.6± 31.57 % Reduction -17.53% -10.22%

0100200300400500600700800900

1000

1 2 3 4

BOD

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Control

Aspergillus niger

Fig. 1. BOD of the tannery effluent before (control) and after biodegradation (72hrs) using fungi and bacteria.

0

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Fig. 2. COD of the tannery effluent before (control) and after biodegradation (72hrs) using fungus and bacterium

325 International Journal of Current Research, Vol. 3, Issue, 4, pp.324-326, April, 2011

tannery in different concentrations (viz 25%, 50%,75% and 100%) using above fungus and bacterium. Two important physicochemical parameters namely BOD and COD were analyzed in the biotreated tannery effluent after 72 hours of incubation. Results of the tannery effluent treated with the fungi Aspergillus niger and Bacterium Bacillus cereus at different concentration (25%, 50%, 75% and 100%) for 72 hrs is shown in the Table 2 & 3. The BOD of tannery effluent before (control) and after bioremediation (72 hrs) by individual fungus and bacteria is shown in Table 2. The COD of tannery effluent before (control) and after biodegradation (72 hrs) by individual fungus and bacteria is shown in Table 3. In all the concentrations of tannery effluent Aspergillus niger was found to reduce BOD and COD to maximum extent followed by the bacterium Bacillus cereus The decrease in BOD and COD values in the biotreated effluent is statistically significant P<0.001.

DISCUSSION

Recent developments in biotechnology now offers opportunities to modify organisms so that their basic biological processes are more efficient and can degrade more complex chemicals and higher volumes of waste materials. Notable accomplishments of the environmental biotechnology include the cleanup of waste water especially from tanneries and other industries. Bioremediation is addressed as one example of an environmental biotechnology. Due to its comparatively low cost and generally benign environmental impact, bioremediation offers an attractive alternative and/or supplement to more conventional clean-up technologies. . Use of microbes to concentrate pollutants is an emerging research area. Microorganisms, bacteria and fungi are nature’s original recyclers. Their capability to transform natural and synthetic chemicals into sources of energy and raw materials for their own growth suggests at hat expensive chemical or physical remediation processes might be replaced or supplemented with biological processes that are lower in cost and more environmentally benign. The present study on tannery effluent from Tiruchirappalli is aimed at analyzing the characteristics to tannery effluent and to reduce certain parameters that pose threat to the environment. In the present investigation, the physico-chemical characteristics of the untreated effluent have revealed that it is acidic with high BOD and COD, organic matter, unpleasant odour and colour. The present study has revealed that high levels of BOD in the tannery effluents indicating high organic load. Present investigation is in agreement with the studies on tannery effluent. (Kulkarni, 1992).

Further the presence of organic matter will promote anaerobic action leading to the accumulation of toxic compounds in the water bodies (Goel, 2000). In the present investigation, high level of COD in the tannery is recorded. Raj et al., 1996 have recorded higher values of COD from the tannery effluent of Chrompet. Further high COD may be due to high amount of inorganic compounds which are not affected by the bacterial decomposition (Nagarajan and Ramachandramoorthy, 2002). Microbes in the environment play an important role in the cycling and fate of organic chemicals and can destroy them through bioremediation. The present study reveals that Bacillus cereus and Aspergillus niger are able to reduce the BOD and COD in all concentrations of the tannery effluent. Studies also reveal that Aspergillus niger was more efficient and has high degrading ability to reduce BOD and COD than Bacillus sp. Hence it may be suggested that fungus will be beneficial for bioremediation and purification of effluents.

REFERENCES APHA, 1989. Standard methods for the examination of water

and waste water. American Public Health Association, Washington, 17th Edition. D.C. pp.1193.

Barnett, H.L. and Hunter, B.B. 1972, Illustrated genera of imperfect fungi. III edition, Burgers publishing company. Minneapolis, Minnesota.

Ellis, M.B. 1971. Dematiaceous Hyphomycetes, common wealth mycologists institute, kew, Surrey, England.

Eye, J.D. and Lawrence, L. 1971. Treatment of wastw from a sole leather industry. J.Wat. pollut. Cont. Fed., 43:2291-2302.

Gilman, J.C. 1967. A manual of soil fungi, oxford and IBH Pub. Co. Calcutta.

Goel, P.K. 2000. Water pollution causes,effects and control. New Age International (P) Ltd., Publ. New Delhi, 269

Jackson, M.C. 1958. Soil chemical analysis, Premise Halls Inc., Englewood cliffs, New Jersey : 498.

Kulkarni, T.T.1992. Source and characteristic of dairy wastes from a medium size effluent on microorganisms plant growth and their microbial change. Life.Sci.Adv.,3:26-78

Raj, E.M., Sankaran, D.P Sreenath, S.K . Kumaran, S. and Mohan, M. 1996. Studies on the treated effluent characteristics of a few tanneries at Chrompet, Madras. Ind, J. Environ.Prot., 6: 252-254

Subramanian, C.V. 1971. Hyphomycetes (An account of Indian species, except cercosporae) ICAR, New Delhi.

326 International Journal of Current Research, Vol. 3, Issue, 4, pp.324-326, April, 2011

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RESEARCH ARTICLE

PERFORMANCE EVALUATION OF COMMERCIAL FEEDS ON FEEDING AND GROWTH OF THE

PRAWN, MACROBRACHIUM ROSENBERGII AT PRAWN FARM, THANJAVUR.

1Sekar. P., 1George John., 1*Balasundaram. A ., 1Krishnamoorthy, R., 1Babunath, R., and 2Aneez Mohamed. M

1 P. G. Research Department of Zoology, Periyar E.V.R. College, Tiruchirappalli-620023, Tamil Nadu, India

2 P. G. Research Department of Zoology, Jamal Mohamed College, Tiruchirappalli-620023, Tamil Nadu, India


Feeding and growth experiments were conducted with reference to three commercial feeds namely, C.P.Scampi feed, Aqua Prime+ feed, and Higashimaru Aqua Feed. The experiments provided evidences to show that C.P.Scampi feed is better than other two commercial feeds under investigation. A body weight of 746 mg/day was gained in those juveniles fed with C.P.Scampi feed, whereas Higashimaru Aqua Feed promoted growth at the rate of 540 mg/day and Aqua Prime + Feed only 296 mg/day. A similar trend was recorded with reference to increase in length (0.0166 mm / day, 0.0141 mm / day, 0.0114 mm / day, and in width 0.13 mm/day, 0.086 mm/day and 0/.064 mm/day).

INTRODUCTION

In the Indian subcontinent, 75% of aquaculture development is on the east coast along the Bay of Bengal. India has recorded a dramatic increase, emerging as the third largest producer of cultured shrimp in the world. In 1991, the country’s aquaculture production, formed approximately 12 to 14% of the total world aquaculture production (Jhingran 1991). Driving forces in aquaculture development are the increasing demand for aquaculture produce, potential for earning foreign exchange, utilization of waste lands for productive purposes, reasonable profits and income, and the urgent need for sustainable food supply. The most important species from the point of commercial aquaculture is the giant long legged freshwater prawn Macrobrachium rosenbergii, commercially known as “Scampi”. It is able to grow both in freshwater and low saline waters. It is compatible for polyculture with carps. Omnivorous in feeding habit, Hardihood and grows fast. It is fairly well resistant to various diseases. Above all it has good consumer preference. It has ever increasing demand both in the domestic as well as export market.To increase freshwater prawn culture activity seed production needs to be enhanced in hatchery and aquaculture. Knowledge in this technique is to be improved by undertaking research on hatchery by local condition. Since seed production requires very delicate optimal conditions, rigorous experimental studies are essential to perfect the technology, suitable for local conditions. *Corresponding author: [email protected]

Hence the present work is undertaken to evaluate the performance of seed production techniques in EMK hatchery unit and to suggest ways and means to improve its efficiency. MATERIALS AND METHODS Macrobrachium rosenbergii is found extensively in the tropical and sub-tropical waters of the Indo-Pacific region in Malaysia, Thailand, Philippines, India, Srilanka, Bangladesh, Myanmar, Indonesia and Vietnam. They are generally found in freshwaters like ponds, rivers, lakes, ditches, canals, depressions, low lying flood plains and river mouths. Most of the species spend their early life in the brackishwater that is directly (or) indirectly connected with the sea. Some species complete their life cycles in freshwater, but these are not of commercial importance. Prawns move upstream, entering lakes and even paddy fields, upto about 200 km from the sea. This type of migration is observed not only in M. rosenbergii but also in other species of Macrobrachium.M. rosenbergii has been used in research more than any other species and has been introduced into many new countries for commercial culture. Breeding biology of Macrobrachium rosenbergii have been studied in the EMK fish and Prawn Seed Farm in Vialur, Thanjavur Dist., Broodstocks (brooders) are maintained in separate tank, periodic examination of males and females were carried out for observing sexual maturity. Observation on breeding biology was made in female for

ISSN: 0975-833X


International Journal of Current ResearchVol. 33, Issue, 3, pp.051-053, March, 2011


Article History:

Received 19th January, 2010 Received in revised form 17th February, 2011 Accepted 1st March, 2011 Published online 13th March, 2011

© Copy Right, IJCR, 2011, Academic Journals. All rights reserved.

Key words: Macrobrachium, prawn feed, Thanjavur.

maturity of eggs. The broodstock with fertilized eggs are selected for hatching purpose. The EMK prawn farm is using the commercially available formulated feed, namely, C.P.Scampi Feed, Aqua Prime+, and Higashimaru Aqua Feed. These commercial feeds were chosen for the evaluation of nutritional value inorder to promote the growth in prawn. For the present study about 25 nos. post larvae were introduced each into three separate fibre glass tank with the capacity of 95 lit. The water quality parameters such as temperature (Cº), pH, Dissolved Oxygen (DO) and Salinity (ppt) were periodically analysed. The post larvae of Macrobrachium rosenbergii were allowed to acclimatize in the laboratory condition for 10 days. After acclimation the post larvae were fed with the three specific commercially formulated feeds with 10% of its body weight. The total body length [tip of Antennae to end of the telson], body weight, and body width was periodically measured once in seven days. The experiment was carried out for 60 days during from Sep. to Nov. 97). The present study to increase the freshwater prawn culture activity, seed production needs to be enhanced in hatchery and aquaculture. The hatchery technology followed in EMK prawn farm is largely based on the techniques explained by Reddy (1997) and Rayes et al. (1985). However, a mass mortality of post larvae was experienced during seed production period under study. The reason for such mortality was attributed to the sudden shift in the salinity of medium of the PL from 14 ppt to freshwater. It was found out that gradual decrease of the salinity of the medium has to be ensured for prevention of mass mortality. The healthy post larvae were sold out. Twenty five post larvae were introduced in the tank of freshwater (Fibre glass tank measuring 54 cm x 46 cm x 30 cm). They were fed with 10% of their body weight of commercial feed. The juveniles were reared for a period of 60 days. The feeding and growth experiments were conducted with reference to three commercial feeds namely, C.P.Scampi feed, Aqua Prime+ feed, and Higashimaru Aqua Feed. The hydrological parameters of the water were examined. The air temperature of the experiment tanks ranged from 27.8°C to 33.6°C and the water temperature from 26.9°C to 30.3°C. The pH fluctuated within a narrow range (7 – 7.5). The dissolved oxygen level fluctuated from 8.6 – 9.0 mg/l, and the salinity did not show wider fluctuation (0.89 ppt to 0.91 ppt). RESULTS AND DISCUSSIONS At the interval of seven days, mean body weight, length, and width were measured in juveniles fed with three commercial feeds (Hossain & Parween 1992). The data are presented in Fig. 1-3, At the end of 60 days the juveniles fed with C.P.scampi feed registered a mean body weight of 4.720 g (initial weight of 0.244 g). Similarly the mean body length increased in 60 days from 4.36 to 11.34 cm, and the body width from 4.00 to 11.8 mm. The juveniles fed with Aqua prime+ feed registered a mean body weight in 60days from 1.900 g (initial weight from 0.120 g). Similarly the mean body length increased in 60 days from 2.82 to 9.70 cm, and the body width from 3.2 to 7.04 mm. The juveniles fed with Higashimaru Aqua Feed registered a mean body weight of 3.460 g (initial weight from 0.216 g) in 60 days. Similarly,

the mean body length increased in 60 days from 3.70 to 10.2 cm, and the body width from 3.4 mm to 8.6 mm. The evaluation experiments provided evidences to show that C.P.Scampi feed is better than other two commercial feeds under investigation. A body weight of 746 mg of body weight / day was gained in those juveniles fed with C.P.Scampi feed, whereas Higashimaru Aqua Feed promoted growth at the rate of 540 mg/day and Aqua Prime + Feed only 296 mg/day. A similar trend was recorded with reference to increase in length (0.0166 mm / day, 0.0141 mm / day, 0.0114 mm / day, and in width 0.13 mm/day, 0.086 mm/day and 0/.064 mm/day). The data on growth rate and amount of food supplied and consumed are used for calculation of food conversion ration with respect to each commercial feed. Feed intake Feed Conversion Ration = ------------- Weight gain The most important aspects of hatchery is the cost-benefit analysis (Shang 1981 & Fuller 1992). In the present

Fig.1. Body weight of juveniles of Macrobrachium rosenbergii in relation to three types of commercial feeds.

746

540

296

0

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200

300

400

500

600

700

800

C.P.Scampi Feed Higashimaru Aqua Feed Aqua Prime+ Feed

Fig.2. Body length of juveniles of Macrobrachium rosenbergii in relation

to three types of commercial feeds.

0.0166

0.0141

0.0114

0

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0.004

0.006

0.008

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Fig.3. Body width of juveniles of Macrobrachium rosenbergii in relation

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052 International Journal of Current Research, Vol. 3, Issue 3, pp.051-053, March, 2011

work annual profit is around 34.7% (Rs.10, 74,100). Hence the annual profit would be substantially increased. However, demand for seeds will not be uniform, unless more prawn culture farms are encouraged to operate by Government Agencies. REFERENCE

Hossain M A and Parween S 1992 Length, weight and width

relationships and condition factor of Macrobrachium

Jhingran V G 1991 Fishes and fisheries of India, IIIrd Pub. Hindustan Publishing Company of India, p 540- 52. malcolmsonii (E.M. Edw.). Fish. Res Lab., Dep. Zool. Rajshahi Univ. Rajshahi- 6205, Vol.24(3):268-69.

Rayes E and Torres P L Jr. 1985 Prawn hatchery design and construction SEAFDECAQD Training material on small scale prawn hatchery operations and management. Tiqbhavan, lloilo: SEAFEDEC Aquaculture Dept., 12p.

Reddy A K 1997 Management of freshwater prawn hatchery and future possibilities. Fishing Chimes., 2: 32- 37.


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RESEARCH ARTICLE

CATION CONCENTRATION IN MUSCLE, GONAD AND HEPATOPANCREAS OF THE FRESHWATER CRAB Paratelphusa hydrodromous

1Sekar, P., 1George John., 1*Balasundaram, A., 1Krishnamoorthy, R., 1Babunath, R and

2Aneez Mohamed, M. 1P.G. Research Department of Zoology, Periyar E.V.R. College, Tiruchirappalli-620023, Tamil Nadu, India 2P.G. Research Department of Zoology, Jamal Mohamed College, Tiruchirappalli-620023, Tamil Nadu, India


This study aimed to estimate the cations (calcium, potassium & sodium) in the fresh water field crab Paratelphusa hydrodromous. The sodium concentration in hepatopancreas gradually decreased are size of the crab increase and there was an inverse relationship between gonad and hepatopancreas in sodium ion concentration. In muscle and hepatopancreas the calcium ion concentration gradually decreased as size group advanced (increase). However in gonad it varied in different size groups and the variation in the calcium ion concentration is may be correlated with moulting cycle of the crab. The potassium ion concentration of P. hydroromous in selected tissues showed slight variation in different size group. However in gonad, hepatopancreas an inverse relationship was noticed.

INTRODUCTION

The fish and fishery products such as prawns, shrimps and crabs are chief sources of protein. Dietary protein deficiency can easily alleviated by consuming fish and fishery product. The superiority of fish and fishery product compared to other protein rich food such as chicken, mutton are suitable for human nutrition. Fish and fishery product is not only easily digestable but also containing all essential amino acids, like lycin, metheonin, histidine, etc., *Corresponding author: [email protected]

Vitamin such as A, B, C, D and B12, minerals like Ca, K, Fe, Na, Mg and Sulphur are present in sufficient quantity. The fishery product especially crabs are having high protein value along with minerals and vitamins. The cations such as Calcium, Potassium, Sodium are also distributed in considerable quantities. Ionic concentrations play an important role in the osmo-ionic regulation of the organisms and some extent in the enzymatic activities of the organisms. The cations are also influence in the normal physiological activities and nervous co-ordination and also in gametes production (Gross 1958; Lockwood 1969).

ISSN: 0975-833X


International Journal of Current ResearchVol. 33, Issue, 3, pp.047-050, March, 2011


Key words: Paratelphusa hydrodromous, Cations, Tissue study.

Article History:

Received 19th January, 2010 Received in revised form 17th February, 2011 Accepted 1st March, 2011 Published online 13th March, 2011

© Copy Right, IJCR, 2011, Academic Journals. All rights reserved.

The ions like Na+, K+, Ca++, and Mg++ are important in the composition of the tissues. The concentration are these ions vary in various tissues during reproductive cycle, nutritive cycles and moulting cycle like pre-moult, post-moult and inter-moult stages of the crab. They also play a vital role in the metabolism of the organism information on the distribution and concentration of cations in the aquatic organisms like fishes, prawns, shrimp and crab is require to assess accumulation of ions in the tissue and possibly transfer for human through food chain (Sather 1967). Hence, the present study aimed to estimate the cation such as calcium, potassium, sodium in the fresh water field crab Paratelphusa hydrodromous. MATERIALS AND METHODS The random collection of freshwater field crab Paratelphusa hydrodromous were made from the field and brought to the laboratory for acclimation. They weighed accurately and the carapace length and breath were recorded. The individuals were grouped into four size groups 10, 13, 15 & 19 gm according to their weight. The male and female were separated and they were sacrified to remove tissues of muscle, gonad and hepatopancreas quickly after dissect ion. The tissues were placed in the Petridisc separately and fixed in 5 percent TCA solution, to stop the enzymatic reaction. These tissues were taken for analysis of cation by using the standard procedure. RESULTS AND DISCUSSION The concentration of sodium, calcium and potassium ion in muscle, gonad and hepatopancreas of four different size groups of Paratelphusa hydrodromous, are summerised in Fig. 1-3. The concentration of sodium ion in the muscle, ranged from 98.6 ppm to 81.4 ppm wet weight and it was found to be maximum lower weight group and minimum. In third size group and it slightly increase in the fourth size group. In gonad the sodium ion concentration ranged from 91.6 ppm to 102.3 ppm wet weight basis. The high value of (102.3) ppm was a recorded in the third size group and lower value (91.6 ppm) were

Fig.1. Sodium concentration of P. hydrodromous in

relation to different size groups

Fig. 2. Calcium concentration of P. hydrodromous in

relation to different size groups

Fig. 3. Potassium concentration of P. hydrodromous

in relation to different size groups recorded in low size group. In gonad the sodium ion concentration slightly increased as size group

0

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Muscle Gonad Hepatopancreas

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Size Group



increased. In hepatopancreas it ranged from 83.7 ppm to 97.5 ppm wet weight. It was found to be high (97.5 ppm) in lower size group and low (83.7 ppm) in higher size group. The sodium concentration in hepatopancreas gradually decreased are size group increase, there was an inverse relationship notice between gonad and hepatopancreas in sodium ion concentration. It success that the transfer of sodium from hepatopancreas to gonad during gonad maturity (Fig. 1). The cations also undergo changes during the reproductive and moulting cycles. Sodium is the most abundant cation in sea water which is known to participate in several developmental processes (Lockwood, 1969; Shaw and Suttliffe, 1961). Sodium is the principal cation of extracellular fluids of most animals. In freshwater field crab Paratelphusa Hydrodromus the sodium level in gonad slightly increased as the size group advanced as in hepatopancrease in the muscle. Similar observation was noticed in sodium concentration of haemolymph in the crab Neptunus pelagicus and Scylla serrata. In these animals the sodium levels are possibly affected by increasing temperature of the medium (Senthilkumar and Desai, 1978). Seasonal variations in haemolymph composition with reference to age, sex and moult cycle of Oreconectus limosus (Andrews, 1967). Carcinus maenas (Greenway, 1976; Shaw and Suttifle (1961) reported that Gammarus duebeni and G. pulex pulex active uptake of sodium is involved in the distribution of this ion between blood and medium. Sodium ion was partially reabsorbed from the urine, in the crab Carcinus maenas. The cation distribution between blood and medium is mainly passive, hyper ion regulation in dilute sea water is affected largely by changes in the rates of uptake (Zander, 1980). The calcium ion concentration in the muscle, ranged from 8.8 ppm to 10.3 ppm on wet weight basis. It was found to be maximum (10.3) ppm in the lower size group and minimum group (8.8 ppm) in higher size group. In gonad the calcium ion concentration varied from 8.3 ppm to 10.3 ppm, third size group and higher value in second size group. In hepatopancreas the sodium ion concentration ranged from 7.0 to 8.7 ppm wet

weight, the value was found to be low (7.0 ppm) in high size group and (8.7 ppm) in the first size group. In muscle and hepatopancrease the calcium ion concentration gradually decreased as size group advanced (increase). However, in gonad it varied in variable to size group the variation in the calcium ion concentration is may be correlated with moulting cycle of the crab (Fig. 2). In depacod crustaceans wide variation of calcium concentration have been reported by many workers. Hormonal control and calcium level in the crab Scylla serrata have been observed (Menon and Sivadass, 1963). Dall (1965) reported that the calcium ion concentration differs, with body size, calcium ion concentration varied in the exoskeleton and tissues during moulting and producting cycles of the shrimp. In the present study calcium concentration of Paratelphus hydroromous gradually decrease in the muscles and hepatopancreas as size group advanced were as in gonad, it vary invariable to size group. The present study agrees with earlier observations in Neptunes pelagicus and Scylla serrats (Senthil kumar and Desai, 1978) Carcinus maenas (Roer, 1980) and Ozioterphusa senex senex (Srinivasulu Reddy, 1985). The potassium in the muscle ranged from 8.1 ppm to 9.3 ppm wet on weight basis. It was found to be maximum (9.3 ppm) in the small size group and minimum (8.1 ppm) in third size group. In gonad the potassium ion concentration ranged from 6.8 to 8.6 ppm wet weight. The lowest values was recorded (6.8 ppm) was recorded in first size group and highest value (8.6 ppm) was recorded in fourth size group. In hepatopancreas it varied from 7.5 to 8.9 ppm wet weight. The potassium ion concentration was found to be minimum (7.5 ppm) in second size group and (8.9 ppm) in first size group (Fig. 3). The potassium ion concentration in various tissues showed slight variation in different size group when compared to the calcium and potassium. Potassium is another important cation, which also influences the osmo-ionic regulation of marine animals (Dehnel and Carfoot, 1965; Skinner 1965; Sather 1967; Poat 1967 and Towle, 1976). The potassium ion concentration in Paratelphus hydroromous in various tissues showed slight variation in different size groups. However in gonad, hepatopancreas inverse


relationship notice in the size group. The present study agrees with earlier observation ( Towle 1991, Neufield (1980) studied the salinity adaptation of gill K+ ATP-ase in the blue crab Callinectus sapidus. According to Moran and Pierce (1984), K+ regulates the salinity tolerance.

REFERENCE

Andrews, P. 1967. Seasonal variation of the haemolymph composition in cray fish Orconectus limosus, Vergl. Physiol., 47(1): 7-43.

Dall, W. 1965. Studies on the physiology of a shrimp Metapenaeus sp. (Crustacea: Decapada: penaeidae) Calcium metabolism. Australian. J. Mar. Freshwat. Res., 16: 181-203.

Dehnel, P. A. and Carfoot, T. H. 1965. Ion exchange in two species of intertidal crabs. Comp. Biochem. Physiol., 15: 377-397.

Greenway P 1976 The regulation of haemolymph calcium concentration of the crab Carcinus maenas (L.), J. Exp. Biol., 64: 149-157.

Gross, W. J. 1958. Potassium and sodium regulation in an interdial crab. Biol. Bull. Woods. Hole., 114: 334-337.

Lockwood, A. P. M. 1969. Aspects of the physiology of crustacean. Oilver and Boyd, Edinburgh and London.

Menon, K. R. and Sivadass, P. 1963. Hormonal control of calcium and sugar levels of the estuarine crab Scylla serrata. Proc. Indian Acad. Sci. B., 67(3): 132 – 140.

Moran, W. M. and Pierce, S. K. 1984. The mechanism of crustacean salinity tolerance, cell volume regulation by K+ and glycine effluxes. Mar. Bio., 81:41-46.

Neufield, G. J. 1980. Salinity adaptation of gill Na, K, ATP-ase in the blue crap Callinectes sapidus. J. Exp. Zool., 211: 215-24.

Poat, P. C. 1967. Cation transport and metabolism as a function of salinity in the excised gill of Carcinus maenas. Comp. Biochem. Physiol., 39: 669-707.

Roer, R. D. 1980. Mechanisms of reabsorption and deposition of calcium in the carapace of the crab Carcinus maenas. J. Exp. Biol., 88: 205-18.

Sather, D. T. 1967. Studies on the calcium and phosphorous metabolism of the crab Podophthalmus wigill (Fabricius). Pacific Science. 21: 193-203.

Shaw, J. and Suttliffe, 1961. Studies on the ionic regulation of Carcinus maenas (L.), I. Sodium balance. J. Exp. Biol., 38: 135-152.

Skinner, D. M. 1965. Physiological salt solution for the land crab. Carcinus lateralis. Biol. Bull., 192: 355-365.

Srinivasulu Reddy, P. 1985. Post-moult calcification of the exoskeleton of Indian Rice field crab. Oziotelphusa senex senex. (Fabricius). Com. Physiol. Ecol., 10 (2): 74-7.

Towle, 1991. Molecular analysis of the electrogenic sodium hydrogen antiporter in crustacean gill. American. Zool., 31(5): 56A.

Zander, I. P. 1980. Regulation of blood ions in Carcinus maenas (L.), Comp. Biochem. Physiol., 65A: 97-108.


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RESEARCH ARTICLE

IMPACT OF LEAD NITRATE ON THE GILLS OF TIGER SHRIMP, PENAEUS MONODON EXPOSED TO SUBLETHAL CONCENTRATIONS LEADING TO HISTOLOGICAL CHANGES

1*Palani Kumar, S., 2Sharadhamma, A.S., 3Swamydoss Daniel, G., and M. Aneez Mohamed4

1Department of Zoology, A.V.V.M., Poondi Pushpam College, Thanjavur, Tamilnadu, India 2Assistant Surgeon, Government primary Health Centre, Thanjavur, Tamilnadu, India 3Controller of Examinations, PRIST University, Vallam, Thanjavur, Tamilnadu, India 4Department of Zoology, Jamal Mohamed College (Autonomous), Tiruchirappalli, Tamilnadu, India


In the present study the tiger shrimp, Penaeus monodon were exposed to three sublethal concentrations of Lead nitrate (1.66, 3.33 and 6.60 mg/L) for a period of 24 hrs,48 hrs,72 hrs,96 hrs respectively. The Gills of the shrimps were then dissected out and processed for light microscopy studies. Exposed shrimps were found to result in several alterations in the histoarchitecture of Gill. The alterations included: necrosis of the epithelial cells of the Gill, hemocytic infiltration in the interlamellar spaces, were observed in the Gills. The results obtained suggest that the Gills of shrimps exposed to sublethal concentrations of Lead nitrate were structurally altered. Such alterations-could affect vital physiological functions, such as absorption, storage and secretion of the Gills, which in turn could ultimately affect the survival and growth of Penaeus monodon.

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INTRODUCTION

Metal accumulation in the environment continuously increases owing to the anthropogenic activities and they tend to concentrate in all the aquatic matrices. The low-dose heavy metal exposure to aquatic organisms may result in various manifestations of biochemical, physiological, and histological alterations in primary tissues (Hinton et al., 1973; Ghate and Mulherkar 1979; White and Rainbow 1986; Kaliamurthy et al.,1994; Yamuna et al., 1996). Heavy metals occur naturally in the environment and are found in varying levels in all ground and surface waters (Martin and Coughtrey, 1982). According to Mason (1991), heavy metal pollution is one of the five major types of toxic pollutants commonly present in surface waters. The important environmental pollutants are those that tend to accumulate in organisms, those that are persistent because of their chemical stability or poor biodegradability, and those that are readily soluble and therefore environmentally mobile (Hellawell, 1986; Sanders, 1997). Histology, the microscopic study of tissue, provides essential knowledge of living cells as the basic building blocks of all living organisms. It examines the structural organisation of these cells in different tissue types allowing better understanding of morphological aspects and physiological *Corresponding author: [email protected]

processes of different organs. According to Short and Meyers (2001), histology is an important field regarding fish health that can often detect subtle conditions or early signs of disease not easily recognized on gross examination. Results from a histological assessment, can provide better insight into the environmental and/or physiological demands presented to fish in their natural environment (Short and Meyers, 2001). Heavy metals including lead are found in various tissues of fish and shrimps (Vazquez et al., 2001).The ultrastructural alterations are suggestive of the operation of compensatory mechanisms within the test prawns to enable it to tolerate Hg toxicity. However, these alterations would have an impact on the cellular integrity of the gills and hepatopancreas and such alterations can be taken as ‘biomarkers’ for assessing Hg pollution in the aquatic environmen (Yamuna et al., 2009). Therefore, it is likely that exposure to a noxious chemical, such as a Lead nitrate, would be reflected in alterations in the structure of the tubules and epithelial cells. Several such structural alterations were noted in the hepatopancreatic tubules of test shrimps, which had been exposed to lead nitrate in the present study. The aim of the present study is to assess the histological changes in the tiger shrimp, Penaeus monodon exposed to sublethal concentrations of Lead nitrate. Which is an economically important species cultured in India. The microscopic study of the tissue provides essential knowledge of living cells as the basic building blocks of all living

ISSN: 0975-833X


International Journal of Current ResearchVol. 3, Issue, 11, pp.045-048, October, 2011


Article History:

Received 5th July, 2011 Received in revised form 8th August, 2011 Accepted 7th September, 2011 Published online 15th October, 2011

Key words:

Histology, Gills, shrimp, Penaeus monodon, Lead nitrate.

organisms. It examines the structural organization of the cells in different tissue types allowing better understanding of morphological aspects and physiological process of different organs. MATERIAL AND METHODS Experimental animals The shrimps were collected from culture pond at Velankanni, Latitude: 10.6833, Longitude: 79.8333, Lat (DMS): 10° 40' 60N, Long (DMS): 79° 49' 60E, Time zone (est) of Nagapattinam District, South India. The shrimps were acclimatized to laboratory condition by using in-door fiber tanks, each with 1.5 m in diameter and 1 m in height, containing 0.80 m water with adequate aeration, and 20% of water volume changed daily. The shrimps were acclimated under the light–dark cycle of 12:12 for two weeks before the experiment. The water temperature was maintained at 27–28 °C. Salinity, 40 ppt; total hardness, 255.0 mg/l; pH, 8.2; nitrate, 1.6 mg/l; chloride, 27.0 mg/l; ammonia, 0.058 mg/l; dissolved oxygen, 6.7 mg/l; BOD, 5.8 mg/l; COD, 14.7 mg/l; and total solid, 1.7 g/l). Commercial shrimp feed was provided daily at 3% of the body weight. Tissue preparation for histological observation The five to eight posterior gills were dissected and immediately fixed in Bouin’s fixative for 48 h. The preserved tissues were processed by a routine histological method (Gurr 1962), dehydrated in an alcohol series, cleared in xylene, infiltrated with liquid paraffin at 58 °C, and finally embedded in paraffin blocks. The blocks were trimmed and sectioned at 5–8 µm thick cut on a rotary microtome (Weswox MT Chenai, India), were stained with Harris' Hematoxylin and counter-stained with Eosin (H&E stain). Then the slides mounted with DPX and observation under a light microscope (Woods and Ellis, 1994).

RESULTS

Control Inter lamellar space normal, No Haemocytes infiltration, Gill tips are normal Cells are normal, Pillar cells are normal, Haemocytes not clear , No cell necrosisiof gill lamellae and cells ,Epithelial cells are normal L, Lamellar epithelium are normal. The gills are well developed nourished and the primary gill filaments are attached to the branchial arch and each supported by an independent gill ray.A number of secondary lamellae are seen attached on both sides of primary filament any they are the main part of gaseous exchange The C.S of gill shows secondary lamellae comes out as fingershaped from the supportive rays .Histologically, the gill shows clearly the occurrence of connective tissues,core cartilage, glandular epithelial cells and filamentous cells excluding arterioles and veinules. The shape of the filament cells are spherical or oblong with a nuclei.The cytoplasm stains more with eosin than cuboidal cells.The cuboidal cells are glandular and responsible for secretion of mucous but the filament cells are essential for respiratory functions (Plate A).

Plate A. Penaeus monodon – Gill (Control)

Plate B. Damage in the basal attachment area (Branchial arch) Cells were damaged and shows sign of deformities SLC-I

Plate C. Lifting of lamellar epithelium Inter lamellar space seen nucleated cells seen SLC-II

Sublethal conc. I (1.66 mg/L) The impact of Lead nitrate shows mild structural changes and damage on the basal attachment of gill. Deformities in the (Branchial arch) basal gill attachment area .The cells show signs of damage and deformities. (Plate B) Sublethal Conc. II (3.33 mg/L) In this concentration the inter lamellar space seen, Lifting of Lamellar epithelium. Necrosis of gill lamellae and cells. Abnormal gill cells and gill tips were damaged. (Plate C)

046 International Journal of Current Research, Vol. 3, Issue, 11, pp.045-048, October, 2011

Plate D. Gill tip damage and deformities in cellular arrangements, necrosis

Plate E. Lacuna seen with abnormal gill cell tip damages – SLC-III

Plate F. Deformed cells showing damage in the gill basal attachment Lead nitrate treated SLC-III

Sublethal Conc. III (6.60 mg/L) Deformities in the (Branchial arch)basal gill attachment area. The cells show signs of damage and deformities.Abnormal gill cells and gill tips were damaged. Vaculation (Lacunae) is seen. Pillar cells damaged. Proliferation of epithelium. Lifting of Lamellar epithelium. Necrosis and erosion of the filaments are seen in the distal ends .The lead nitrate has impact over the gill cells. Primary gill filaments are attached to the .branchial

arch damaged. Extensive damage of gill structure and cellular alterations are observed. (Plate D) DISCUSSION Following exposure the gills exhibited rapid alterations that include detachment and lifting of the epithelial linings from the surfaces of the gill filament (primary, PL) and respiratory (secondary, SL) lamellae. This leads to extensive haemorrhage from the gills. Thus the quantity of blood flowing across the gills decreased substantially. Simultaneously, uncontrolled regeneration of the PL and SL occured, leading to extensive hyperplasia of the epithelial cells lining the PL, and SL. Consequently, the gill filaments appeared as a cylindrical solid mass of cells with very little or almost no free surface left on the SL for gaseous exchange. The goblet mucous cells also exhibited periodic fluctuations in their density and staining behaviour. The chloride cells showed periodic fluctuation in their number at different stages of exposure. The density of the chloride cells is inversely proportional to the thickness of the epithelial lining of the PL and SL. Due to prolonged exposure, the neighbouring SL fused together and the entire gills appeared as solid mass of undifferentiated cells. Subsequently, the ladder-like arrangement of the pillar cells-blood capillaries of the gills also collapsed, causing asphyxiation and the death of the fish.Similarities observed and coincides with Ram Sanehi Parashar, and Tarun Kumar Banerjee, (2002). CONCLUSION

Thus, the changes observed in the shrimps exposed to 1.66,3.33,6.60 mg/L lead nitrate in the present study were more likely to have represented a progressive loss of basic biological functions of the Gills and it ultimately affect the survival and growth of Penaeus monodon. REFERENCES Ghate, HV and Mulherkar, L. 1979. Histological changes in

the gills of two freshwater prawn species exposed to copper sulphate. Indian J Exp Biol., 17:838–840.

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Hinton, DE., Kemdall, MW, and Silver, BB,. 1973. Biological methods for the assessment of water quality. ASTM STP 528, USA, 194 pp

Kaliamurthy, J., Bhavan, PS. and Geraldine, P. 1994. Alterations in the protein profile and microstructure of gills and hepatopancreas in the prawn, Macrobrachium lamarrei following exposure to hospital wastes. Proceedings of the International Conference on Agrotechnology in the Commonwealth Focus for the 21st Century, Singapore National University, Singapore, pp 121–124

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Ram Sanchi Parashar and Tarun Kumar Banerjee, 2002. Toxic impact of lethal concentration of Lead nitrate on the gills of air-breathing catfish Heteropneustes fossilis (Bloch),Veterinarski Arhiv, 72 (3) : 167 – 183.

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Short, S. and Meyers, T.R. 2001. Histology for Finfish. NWFHS Laboratory Procedures Manual. Version 1 .0. Alaska Fish and Game C.F. Division.

Vazquez, F.G., Sharma V.K., Mandoza Q.A. and Hernandez R. 2001. Metal content of the Gulf of California Blue Shrimp Litopenaeus stylirostris (Stimpson). Bull Environ Contam. Toxicol., 67, 756-762.

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Yamuna, A., Kabila, V. and Geraldine, P. 1996. Biochemical and histological alterations in the prawn Macrobrachium lamarrei following exposure to automobile discharge. Geo J., 40:233–237.

Yamuna1,A., Saravana Bhavan, P. and P. Geraldine1. 2009. Ultrastructural observations in gills and hepatopancreas of prawn Macrobrachium malcolmsonii exposed to mercury., Journal of Environmental Biology. September 2009, 30(5) 693-699 .www.jeb.co.in.

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