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Abstract
THESIS ABSTRACT
Effect of Black seed (Nigella sativa) on the Immune Status of Catfish (Clarias gariepinus)
Abeer Ezzat Mahmoud Ali
B.V.Sc. Assiut University 1992
Diploma of Fish Diseases Assiut University 1994
M.V.Sc. Assiut University 2000
Supervised By
Prof. Dr. Shaban M. Ahmed
Prof. Dr.Imam A.A. Mekkawy
Prof. Dr. Abu El-Magd M.Mohamed
The aim of this study is to investigate the effect of black seed Nigella sativa on the immune status on Nile catfish, Clarias gariepinus, evaluate the effect of Nigella sativa on the haematological parameters of Clarias gariepinus and evaluate the overall effect of Nigella sativa as immunostimulant in case of Aeromonas hyrophila infection. During present experiments, 160 healthy Clarias gariepinus with an average weight 250 50gm. were collected from the River Nile and El-Ibrahemia canal in Assiut City. Fish have been divided into seven groups, each of twenty five fish except the seventh group was ten fish, group I [control], group II [injected intraperitoneal with saline solution (0.65 % NaCL)], group III [injected i.p. with 0.05ml Nigella sativa oil/100g fish], group IV [injected i.p. with 0.1ml Nigella sativa oil/100g fish], group V [injected i.p. with 0.2ml Nigella sativa oil/100g fish], group VI [injected i.p. with 0.3ml Nigella sativa oil/100g fish] and group VII [auto control]. After three weeks, all the first six groups were intraperitoneally challenged by Aeromonas hyDROPhila isolate (0.2ml of 2 107 bacterial cells/ml3/100g fish). Three fish were sacrificed weekly from each of the first sex groups [i.e. at the end of the 1st, 2nd and 4th weeks], where the haematological, biochemical, and histopathological investigations have been carried out.
In the present study, the fish injected i.p. with the highest doses of Nigella sativa oil (0.2ml and 0.3ml/100g fish), showed ascites and signs of peritonitis and the peritoneal cavity was full with bloody fluids and adhesions. All viscera were severly congested but all the fish were appeared normal in gross appearance and in postmortum examination at the end of the 1st week.
The haematological changes in the blood of Clarias gariepinus induced post the i.p. injection of Nigella sativa oil by one week were characterized by a insignificant increase in the total number of RBCs, haemoglobin concentration and PCV values in group III and group IV, while there were significant decrease in the RBCs counts, haemoglobin concentration and PCV values in group V and group VI. At the end of the 2nd week, all these blood parameters were nearly normal except the total counts of RBCs which were normal in groups III and IV, while increased in group V and VI. Post-bacterial infection by one week [i.e. at the end of the 4th week], the haematological changes in the blood were characterized by decrease in the total number of RBCs, haemoglobin concentration and PCV values in all living groups (groups III, IV, V and VI).
The mean cell volume (MCV) values were significantly increased post-injection with Nigella sativa oil by one week in groups III, V and VI. The mean cell haemoglobin (MCH) values were variable in all groups. While the mean cell haemoglobin concentration (MCHC) values were insignificantly decreased at the end of the 1st week post i.p. injection with Nigella sativa oil in group III and group IV, at the end of 2nd week in groups V and VI and in all groups post-bacterial infection, while, MCHC values were significantly decreased after i.p. injection with Nigella sativa oil by one week in groups V & VI.
The total leucocyte counts were increased at the end of the 1st & 2nd weeks in groups III & IV, while they decreased in groups V & VI. After the 4th week, the WBCs counts were also increased in groups III & IV and became normal in group V, while they still decreased than normal in group VI.
Evaluation the effect of Nigella sativa oil on the protein fractions revealed that the value of the total protein, albumin and globulin in the serum of Clarias gariepinus were fluctuated along the experimental period. Also, in the studying the effect of Nigella sativa oil on the serum electrolytes, we observed that the analysis of the fish serum for determination of the levels of Na+, K+ and Cl-, showed fluctuated results.
In investigation the effect of Nigella sativa oil on the electrophoretic patterns of the catfish serum refered also fluctuated results.
The microscopic examination of fish injected with Nigella sativa oil revealed, that Nigella sativa has immunostimulant effects which represented by activation of haemopoietic tissues of the spleen and kidney with proliferation of the melanomacphage centers. This effect was more prominent in lower dose (0.05ml Nigella sativa oil / 100g fish). However, hepato-renal damages in the from of degernerative changes in both hepatocytes and renal tubular epithelium were also recorded only in higher doses. Moreover, after challenge with Aeromonas. hyDROPhila, the severity of recorded histopathological changes was varied from mild changes in lower doses and sever in higher doses particularly in the liver where hepato-portal congestion, hepatic degeneration and necrosis with hemosiderosis were observed.
This study support the traditional use of Nigella sativa when used in lower doses [0.05ml Nigella sativa oil/100g fish] and indicate a relative toxicity of this plant extract in higher doses.