Impact of environment changes induced immunological modulation in the fresh water fis
#1

PRESENTED BY:
S. Binu Kumari and H.Kavitha

[attachment=10812]
Abstract
Fish culturists are very aware that a variety of environment stressors including pesticide contaminated run off waters affect the immune system (Anderson, 1990).
Pesticides induced stress causes elevation in blood glucose, corticosteroid hormones and catacholamines that have been linked to impairment of immune and circulating functions and to lowered disease resistance (Melamed et al., 1999). Knowledge of how a pesticide might affect the immunology of a fish species can help in development and management tactics that lessen the impact of pesticide induced stress or even in the development of therapeuties.
Keywords:
Immunesystem, Perticicde, Channa striata.
Introduction:
Teleost fish show analogy with other vertebrates concerning humoral, cellular and non – specific responses (Manning, 1994). The study of immune system of fish is thus having a great impetus both for a better knowledge of the evoluation of the vertebrate immune system and to improve the control of fish health in aquaculture channa striata is a commonly available, commercially valuable south- Indian air- breathing fish. The fish is frequently exposed to pesticide polluted waters (Natarajan et al., 1998). Knowledge on the effect of commonly used pesticide, Metasystox, on the immune system of fish will provide information in understanding the various specific and non-specific defense reaction which will in turn pave the way for the development of effective therapeutic methods.
Material and Methods:
Snake head of species channa striata (Weight: 20-25g; length10-13cm)
Were purchased and allowed to acclimatize to the laboratory conditions for 15 days before using in the experiments. Technical grade metasystox (Oxydemeton-methyl, 0, 0-dimethyl – S-2 (ethyl sulfinyl) ethyl phosphorothioate) of 95 % purity was obtained from Bayer LTD, Bombay. A stock solution of Metasystox was made in acetone (l mg/ml) and suitable quantities of this solution were added to the aquaria to obtain the desired concentrations. Lethal (Lc 50 / 48hr) concentration (5mg / 1) was calculated by a probit method (Finney, 1964) and approximately 1/3 of the LC 50 / 48 hr concentration (1.7 mg / 1) was chosen for sublethal exposure. At this concentration, the fish survived even after prolonged periods of exposure (Natarajan, 1984). Fish in batches of 12 were exposed to sublethal concentration of 1.7 mg/1 for 15 and 30 days.
The test solutions were renewed every 24 hr. Twelve snake head used as controls, were kept in clean tap water with the same concentration of acetone. The blood from the control and Metasystox exposed fishes was collected into 1 ml syringe and 24 gause needle previously rinsed with 10 % sodium heparin. The blood was centrifuged (500 x 5g, 5 min) and the plasma obtained was stored frozen along with serum and mucus samples at – 20oC.
The lysozyme activity in plasma was measured in 1 % agarose(BDH) using a heat killed Micrococcus lysodeikticus (BDH, M. 2500) to perform a standard calibration curve. The ability of fish phagocytes to ingest particles is done with an in vitro phagocytosis assay (Duda, 1996). Phagocyctes from the head kidney were stimulated with phorbol 12-myristate13-enhanced CL method (Scott and Klesius, 1981). The capability of head kidney neutrophils to move was assayed by a migration under agarose technique described by Nelson et al., (1975). The amount of total IgM and specific antibovine y – globulin (BCG) antibody in the snake head plasma was measured by an Enzyme Linked Immuno Sorbent Assay (ELISA) described by Aaltonen et al., (1994). The enzyme linked immunospot assay was used for the enumeration of total immunoglobulin secreting cells (ISC) and antigen (BCG) –specific antibody secreting cells (ASC) in the spleen and the blood (Aaltonen et al., Averages for each measurement of each immunological characteristic of control and experimental groups were compared by one-way ANOVA. If significant results were found, student ‘t’ test was used to determine which specific means differed from the others.
Results and Discussion:
It is evident from the tables 1-5 that Metasystox modulated the immunological parameters significantly at all times of exposure. The lysozyme activity increased in the spleen, kidney, plasma and mucus. The number of yeast cells phagocytized increased significantly in the 15 days treatment. However, in the 30 days exposure. Greater accumulation of IgM was evident at 30 day (+80.90%; P < 0.05) treatment. The anti BGG specific antibodies accumulated significantly at 15 day (+30.63 ; P < 0.05) and 30 day (+36.25 %; P<0.05) treatment.
Metasystox exposure induces both immunosuppression and immunostimulation in channa striata. Lysozyme is a leucocyte enzyme important in inflammation and bacterial killing. Mock and peters (1990) found that plasma lysozyme levels increased in rainbow trout, Onchorhynchus mykiss exposed to a 30 min. confinement stress. While lysozyme is a generally recognized as being part of the humoral non-specific defense system it could be argued that it emanates from phagocytes. The present study confirms that Metasystox include increase in the activities of both mucus and serum lysozyme perhaps providing a defense against pesticide stress. Microphages and the cells of reticulo-histicyte-system cells participate in phagocytosis and in immunogensis in fish (Schaperclaus, 1991).
Suppression of phagocytic activity in the 30 day treatment in turn may lower the resistance of the fish to bacterial infection (Kakuta, 1997). Metasystox causes moderate accumulation of plasma lg profile and anti-BGG specific antibodies. This is an indication of pesticide stress response. Similar findings were reported for bleaching factory effluents (Aaltonen et al., 2000). The number of ISC and the blood ASC response have proved to be sensitive indicator of metasysstox stress. Greater reduction in circulating lg titer after 15 day pesticide exposure indicates that, initially metasystox seriously impairs the immune system of C. striata. However the subsequent increase in lg titer suggest that after initial inhibitions of immune functions, C.striata start to activate some protective or compensatory mechanism. This may be correlated to the capacity of lymphocytes to synthesize metallothionenins in response to exposure of Metasystox.
Metasystox, probably interacts with disulfide groups of immunoglobulins, causing conformational modifications and thus interfering with the antigen-antibody bond in the ELISA. Conformational modifications may also cause acclerated turnover of immunoglobulins.
The lg of telecosts posses the same functions as the lg of mammals against the pathogenic agents (Manning, 1994). However, fish are more susceptible to bacterial infection than terrestrial mammals, because the aquatic environment is a rich source of exposure suppress immune function rendering C.striata easily vulnerable to diseases and there by causing huge economic losses to fish culturists.
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#3
(23-03-2011, 11:45 AM)seminar class Wrote: PRESENTED BY:
S. Binu Kumari and H.Kavitha


Abstract
Fish culturists are very aware that a variety of environment stressors including pesticide contaminated run off waters affect the immune system (Anderson, 1990).
Pesticides induced stress causes elevation in blood glucose, corticosteroid hormones and catacholamines that have been linked to impairment of immune and circulating functions and to lowered disease resistance (Melamed et al., 1999). Knowledge of how a pesticide might affect the immunology of a fish species can help in development and management tactics that lessen the impact of pesticide induced stress or even in the development of therapeuties.
Keywords:
Immunesystem, Perticicde, Channa striata.
Introduction:
Teleost fish show analogy with other vertebrates concerning humoral, cellular and non – specific responses (Manning, 1994). The study of immune system of fish is thus having a great impetus both for a better knowledge of the evoluation of the vertebrate immune system and to improve the control of fish health in aquaculture channa striata is a commonly available, commercially valuable south- Indian air- breathing fish. The fish is frequently exposed to pesticide polluted waters (Natarajan et al., 1998). Knowledge on the effect of commonly used pesticide, Metasystox, on the immune system of fish will provide information in understanding the various specific and non-specific defense reaction which will in turn pave the way for the development of effective therapeutic methods.
Material and Methods:
Snake head of species channa striata (Weight: 20-25g; length10-13cm)
Were purchased and allowed to acclimatize to the laboratory conditions for 15 days before using in the experiments. Technical grade metasystox (Oxydemeton-methyl, 0, 0-dimethyl – S-2 (ethyl sulfinyl) ethyl phosphorothioate) of 95 % purity was obtained from Bayer LTD, Bombay. A stock solution of Metasystox was made in acetone (l mg/ml) and suitable quantities of this solution were added to the aquaria to obtain the desired concentrations. Lethal (Lc 50 / 48hr) concentration (5mg / 1) was calculated by a probit method (Finney, 1964) and approximately 1/3 of the LC 50 / 48 hr concentration (1.7 mg / 1) was chosen for sublethal exposure. At this concentration, the fish survived even after prolonged periods of exposure (Natarajan, 1984). Fish in batches of 12 were exposed to sublethal concentration of 1.7 mg/1 for 15 and 30 days.
The test solutions were renewed every 24 hr. Twelve snake head used as controls, were kept in clean tap water with the same concentration of acetone. The blood from the control and Metasystox exposed fishes was collected into 1 ml syringe and 24 gause needle previously rinsed with 10 % sodium heparin. The blood was centrifuged (500 x 5g, 5 min) and the plasma obtained was stored frozen along with serum and mucus samples at – 20oC.
The lysozyme activity in plasma was measured in 1 % agarose(BDH) using a heat killed Micrococcus lysodeikticus (BDH, M. 2500) to perform a standard calibration curve. The ability of fish phagocytes to ingest particles is done with an in vitro phagocytosis assay (Duda, 1996). Phagocyctes from the head kidney were stimulated with phorbol 12-myristate13-enhanced CL method (Scott and Klesius, 1981). The capability of head kidney neutrophils to move was assayed by a migration under agarose technique described by Nelson et al., (1975). The amount of total IgM and specific antibovine y – globulin (BCG) antibody in the snake head plasma was measured by an Enzyme Linked Immuno Sorbent Assay (ELISA) described by Aaltonen et al., (1994). The enzyme linked immunospot assay was used for the enumeration of total immunoglobulin secreting cells (ISC) and antigen (BCG) –specific antibody secreting cells (ASC) in the spleen and the blood (Aaltonen et al., Averages for each measurement of each immunological characteristic of control and experimental groups were compared by one-way ANOVA. If significant results were found, student ‘t’ test was used to determine which specific means differed from the others.
Results and Discussion:
It is evident from the tables 1-5 that Metasystox modulated the immunological parameters significantly at all times of exposure. The lysozyme activity increased in the spleen, kidney, plasma and mucus. The number of yeast cells phagocytized increased significantly in the 15 days treatment. However, in the 30 days exposure. Greater accumulation of IgM was evident at 30 day (+80.90%; P < 0.05) treatment. The anti BGG specific antibodies accumulated significantly at 15 day (+30.63 ; P < 0.05) and 30 day (+36.25 %; P<0.05) treatment.
Metasystox exposure induces both immunosuppression and immunostimulation in channa striata. Lysozyme is a leucocyte enzyme important in inflammation and bacterial killing. Mock and peters (1990) found that plasma lysozyme levels increased in rainbow trout, Onchorhynchus mykiss exposed to a 30 min. confinement stress. While lysozyme is a generally recognized as being part of the humoral non-specific defense system it could be argued that it emanates from phagocytes. The present study confirms that Metasystox include increase in the activities of both mucus and serum lysozyme perhaps providing a defense against pesticide stress. Microphages and the cells of reticulo-histicyte-system cells participate in phagocytosis and in immunogensis in fish (Schaperclaus, 1991).
Suppression of phagocytic activity in the 30 day treatment in turn may lower the resistance of the fish to bacterial infection (Kakuta, 1997). Metasystox causes moderate accumulation of plasma lg profile and anti-BGG specific antibodies. This is an indication of pesticide stress response. Similar findings were reported for bleaching factory effluents (Aaltonen et al., 2000). The number of ISC and the blood ASC response have proved to be sensitive indicator of metasysstox stress. Greater reduction in circulating lg titer after 15 day pesticide exposure indicates that, initially metasystox seriously impairs the immune system of C. striata. However the subsequent increase in lg titer suggest that after initial inhibitions of immune functions, C.striata start to activate some protective or compensatory mechanism. This may be correlated to the capacity of lymphocytes to synthesize metallothionenins in response to exposure of Metasystox.
Metasystox, probably interacts with disulfide groups of immunoglobulins, causing conformational modifications and thus interfering with the antigen-antibody bond in the ELISA. Conformational modifications may also cause acclerated turnover of immunoglobulins.
The lg of telecosts posses the same functions as the lg of mammals against the pathogenic agents (Manning, 1994). However, fish are more susceptible to bacterial infection than terrestrial mammals, because the aquatic environment is a rich source of exposure suppress immune function rendering C.striata easily vulnerable to diseases and there by causing huge economic losses to fish culturists.

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#4
Impact of environment changes induced immunological modulation in the fresh water fis

Abstract
Fish culturists are very aware that a variety of environment stressors including pesticide contaminated run off waters affect the immune system (Anderson, 1990).
Pesticides induced stress causes elevation in blood glucose, corticosteroid hormones and catacholamines that have been linked to impairment of immune and circulating functions and to lowered disease resistance (Melamed et al., 1999). Knowledge of how a pesticide might affect the immunology of a fish species can help in development and management tactics that lessen the impact of pesticide induced stress or even in the development of therapeuties.
Keywords:
Immunesystem, Perticicde, Channa striata.
Introduction:
Teleost fish show analogy with other vertebrates concerning humoral, cellular and non – specific responses (Manning, 1994). The study of immune system of fish is thus having a great impetus both for a better knowledge of the evoluation of the vertebrate immune system and to improve the control of fish health in aquaculture channa striata is a commonly available, commercially valuable south- Indian air- breathing fish. The fish is frequently exposed to pesticide polluted waters (Natarajan et al., 1998). Knowledge on the effect of commonly used pesticide, Metasystox, on the immune system of fish will provide information in understanding the various specific and non-specific defense reaction which will in turn pave the way for the development of effective therapeutic methods.
Material and Methods:
Snake head of species channa striata (Weight: 20-25g; length10-13cm)
Were purchased and allowed to acclimatize to the laboratory conditions for 15 days before using in the experiments. Technical grade metasystox (Oxydemeton-methyl, 0, 0-dimethyl – S-2 (ethyl sulfinyl) ethyl phosphorothioate) of 95 % purity was obtained from Bayer LTD, Bombay. A stock solution of Metasystox was made in acetone (l mg/ml) and suitable quantities of this solution were added to the aquaria to obtain the desired concentrations. Lethal (Lc 50 / 48hr) concentration (5mg / 1) was calculated by a probit method (Finney, 1964) and approximately 1/3 of the LC 50 / 48 hr concentration (1.7 mg / 1) was chosen for sublethal exposure. At this concentration, the fish survived even after prolonged periods of exposure (Natarajan, 1984). Fish in batches of 12 were exposed to sublethal concentration of 1.7 mg/1 for 15 and 30 days.
The test solutions were renewed every 24 hr. Twelve snake head used as controls, were kept in clean tap water with the same concentration of acetone. The blood from the control and Metasystox exposed fishes was collected into 1 ml syringe and 24 gause needle previously rinsed with 10 % sodium heparin. The blood was centrifuged (500 x 5g, 5 min) and the plasma obtained was stored frozen along with serum and mucus samples at – 20oC.
The lysozyme activity in plasma was measured in 1 % agarose(BDH) using a heat killed Micrococcus lysodeikticus (BDH, M. 2500) to perform a standard calibration curve. The ability of fish phagocytes to ingest particles is done with an in vitro phagocytosis assay (Duda, 1996). Phagocyctes from the head kidney were stimulated with phorbol 12-myristate13-enhanced CL method (Scott and Klesius, 1981). The capability of head kidney neutrophils to move was assayed by a migration under agarose technique described by Nelson et al., (1975). The amount of total IgM and specific antibovine y – globulin (BCG) antibody in the snake head plasma was measured by an Enzyme Linked Immuno Sorbent Assay (ELISA) described by Aaltonen et al., (1994). The enzyme linked immunospot assay was used for the enumeration of total immunoglobulin secreting cells (ISC) and antigen (BCG) –specific antibody secreting cells (ASC) in the spleen and the blood (Aaltonen et al., Averages for each measurement of each immunological characteristic of control and experimental groups were compared by one-way ANOVA. If significant results were found, student ‘t’ test was used to determine which specific means differed from the others.
Results and Discussion:
It is evident from the tables 1-5 that Metasystox modulated the immunological parameters significantly at all times of exposure. The lysozyme activity increased in the spleen, kidney, plasma and mucus. The number of yeast cells phagocytized increased significantly in the 15 days treatment. However, in the 30 days exposure. Greater accumulation of IgM was evident at 30 day (+80.90%; P < 0.05) treatment. The anti BGG specific antibodies accumulated significantly at 15 day (+30.63 ; P < 0.05) and 30 day (+36.25 %; P<0.05) treatment.
Metasystox exposure induces both immunosuppression and immunostimulation in channa striata. Lysozyme is a leucocyte enzyme important in inflammation and bacterial killing. Mock and peters (1990) found that plasma lysozyme levels increased in rainbow trout, Onchorhynchus mykiss exposed to a 30 min. confinement stress. While lysozyme is a generally recognized as being part of the humoral non-specific defense system it could be argued that it emanates from phagocytes. The present study confirms that Metasystox include increase in the activities of both mucus and serum lysozyme perhaps providing a defense against pesticide stress. Microphages and the cells of reticulo-histicyte-system cells participate in phagocytosis and in immunogensis in fish (Schaperclaus, 1991).
Suppression of phagocytic activity in the 30 day treatment in turn may lower the resistance of the fish to bacterial infection (Kakuta, 1997). Metasystox causes moderate accumulation of plasma lg profile and anti-BGG specific antibodies. This is an indication of pesticide stress response. Similar findings were reported for bleaching factory effluents (Aaltonen et al., 2000). The number of ISC and the blood ASC response have proved to be sensitive indicator of metasysstox stress. Greater reduction in circulating lg titer after 15 day pesticide exposure indicates that, initially metasystox seriously impairs the immune system of C. striata. However the subsequent increase in lg titer suggest that after initial inhibitions of immune functions, C.striata start to activate some protective or compensatory mechanism. This may be correlated to the capacity of lymphocytes to synthesize metallothionenins in response to exposure of Metasystox.
Metasystox, probably interacts with disulfide groups of immunoglobulins, causing conformational modifications and thus interfering with the antigen-antibody bond in the ELISA. Conformational modifications may also cause acclerated turnover of immunoglobulins.
The lg of telecosts posses the same functions as the lg of mammals against the pathogenic agents (Manning, 1994). However, fish are more susceptible to bacterial infection than terrestrial mammals, because the aquatic environment is a rich source of exposure suppress immune function rendering C.striata easily vulnerable to diseases and there by causing huge economic losses to fish culturists.
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