Biochemical structure and antibacterial activity of hemolymph in male and female of fresh water prawn (Macrobrachium nipponense (
Subject Areas : Journal of Comparative PathobiologyKarimzadeh, K.*, Zahmatkesh, A., Pormehr, M. . 1
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Keywords: antibacterial activity, hemolymph, electrophoresis, disk diffusion,
Abstract :
The aim of this study was to characterize the biochemical structure and bactericidal activity of hemolymph Microbrachium in different doses on several human bacterial pathogenFirst 80 individual (40 for male and 40 for female) of prawn (M. nipponense) with an average length of 8.1 ± 0.12 were collected from Anzali wetland. After separation of males and female from each other, hemolymph of samples were taken from ventral sinus of the prawn and structure of collected hemolymph was determined through an electrophoresis and FTIR analysis.The antibacterial activity of M. nipponense hemolymph was investigated against five pathogenic bacteria (Vibrio cholera, Klebsiella pneumonia, Bacillus subtilis, Staphylococcus aureus, Escherichia coli) in doses of 25 to 100 micrograms per liter by disk diffusion method.The electrophoretic pattern of males and females hemolymph revealed presence of proteins with molecular weights between 22-100 Kda. Hemolymph structure contained both secondary regular and irregular protein structures by FTIR analysis. Different doses of hemolymph, indicated inhibitory activity against different bacterial strains. Also significant difference was observed between male and female hemolymph in case of antibacterial activity p <0.05).Highest inhibitory effect of hemolymph was shown against Staphylococcus aureus, Bacillus subtilis and Vibrio cholera in male. While the lowest level of antibacterial activity was recorded in female against Bacillus subtilis and Escherichia coli with zone of inhibition 5.9 ± 0.12 and 6.5 ± 0.08 mmResults of this study indicated that fresh water prawn hemolymph was shown well bactericidal activity and can be a substitute for antibiotic-based chemical drugs.
1. Baure, A.W., Kirby, W.M.M., Sherris, M.J.C., Turck, M. (1996): Antibiotic susceptibility testing by a standardized single disc method. Am J Clin Pathol. 45:493-49.
2. Bradford, M.M. (1976): A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal .Biochem. 72:248-54.
3. Chisholm, J.R.S, Smith, V.J. (1992): Antibacterial activity in the hemocytes of the shore crab, Carcinus maenas. J. Ma.r Biol .Assoc. U.K. 72: 529-42.
4. Chodsang, S., Benjakul, S., Visessanguan, W., Hara, K., Yoshida, A., Liang, X. (2012): Low molecular weight trypsin from hepatopancreas of freshwater prawn (macrobrachium rosenbergii): characteristics and biochemical properties. J. Food. Chem. 134:351-8.
5. Cuthbertson, B. J, Bullesbach, E.E., Gross, P.S. (2006): Discovery of synthetic penaeid in activity against antibiotic resistant. Chem. Biol .Drug Des .68:120-7.
6. De'Grave,S., Ghane, A. (2006) :The establishment of the Oriental River Prawn, Macrobrachium nipponense (de Haan, 1849) in Anzali Lagoon, Iran. Aquat. Invasions. 1(4):204-208.
7. Destoumieux, D., Bulet, P., Loew, D., Dorsselaer, V. A., Rodriguea, J., Bachere, E. (1997): Penaedine a new family or antimicrobial peptides isolated from the shrimp penaeus vannamei(Decapoda). J. Biol. Chem.; 72: 28398-406.
8. Hajirasouli, M., Pazooki,J. (2014) :Antimicrobial potential of haemolymph and hepatopancreas of portunussegnis crabs . Int. J .Pharm. Pharm .Sci. 6(8): 601-603.
9. Kawababa, S.I., Nagayama, R., Hirata, M., Shigenaga, T., Agarwala, K.L., Saito, T. (1996):Tachykinin, a small granular component in horseshore crab hemocytes is an antimicrobial protein with chitin-binding activity. J. Biochem. 120:1253-60.
10. Kim, J.K., Kraemer G.P., Neefus, C.DChung. I.K., Yarish, C. (2007): Development and iologicalactivities of marine derived bioactive peptides. J. Funct. Foods. 2(1): 1-9.
11. Laemmle, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680-685.
12. Li, Y., Su, X.R., Li, T.W. (2005): Study on antimicrobial peptides from Bullacta exarata. J. Ocean Taiwa. 24: 145-149.
13. Loker, E.S., Adema, C.M., Zhang, S.M., Kepler, T.B. (2004): Invertebrate immune systems—not homogeneous, not simple, not well understood. Immunol Rev. 198:10–24.
14. Miyata, T., Tokunaga, F., Yoneya, T., Yoshikawa, K., Iwanaga, S., Niwa, M., Takao, T., Shimonishi, Y. (1998): Antimicrobial peptides, isolated from horseshoe crab hemocytes, tachyplesinmII and polyphemisinsI and II:chemical structures and biological activity. J Biochem .106:663-8.
15. Noga, E.J., Arroll, T.W., Fan, Z. (1996): Specificity and some physiochemical characteristics of the antibacterial activity from the blue crabs Callinectes sapidus. Fish Shellfish Immunol . 6(6):403-12.
16. Priya, E.R., Kohilam , Ravichandran S. (2015):Antimicrobial activity from the hemolymph of the hermit crab Clibanarius clibanarius (Herbst 1791). World. J. Fish. Mar .Sci . 7 (4): 263-267.
17. Rameshkurmar, G.S., Ravichandran, M., Kaliyavarathan, G., Ajithkumar,T.T. (2009) :Antimicrobial peptide from the crab, Thalamita crenata(Latreille, 1829). World J. Fish Mar Sci. 1(2):74-9.
18. Rameshkumar, G., AravindhanT., Ravichandran, S. (2009) :Antimicrobial proteins from the crab Chrybdislucifera (Fabricius, 1798). Middle East J of Sci Res .4(1):40-3.
19. Ravichandran, S.S., Wahidulla, L. D., Souza, Rameshkumar, G. (2010) Antimicrobial lipids from the hemolymph of brachyuran crabs. J. Appl. Biochem. Biotechnol .162(4):1039-51.
20. Ravichandran, S., Jeyalakshmi , S., Sudha, S., Anbuchezhian, R.(2010): Antimicrobial peptides from the haemolymph of the prawn Macrobrachium rosenbergii. Bangladesh J Pharmocol . 5: 62–67
21.Ravichandran, S., Sivasubramanian, K., Anbuchezhian, R.M. (2010): Antimicrobial activity the hemolymph of crab Ocypoda macrocera. J. Wor. Appl. Sci. 11: 578-581
22.Smith V.J., Chisholm, J.R. (2001): Antimicrobial protein in crustaceans. J. Adv .Exp. Med. Biol. 484:95-112.
23.Soundrapandian, P. (2009): Nutritive value of hard and soft shell crabs of Portunus sanguinolentus (Herbst). Int J Anim Vet Adv. 1(2): 44-48.
24. Soundarapandian, P., Ananthan, G. (2008): Effect of unilateral eyestalk ablation and diets on the biochemical composition of commercially important juveniles of Macrobrachium malcomsonii (H. Milne Edwards). Int. J. Zool. Res. 4(2): 106-112.
25.Stewart, J. E., Zwicker, B. M. (1972): Natural and induced bactericidal activities in the hemolymph of the lobster; Homarus americanus: products of hemocyte-plasma interactions. Can. J. Microbiol. 18:1499–1509.
26.Tincu, J.A., Taylor, S.W. (2004): Antimicrobial peptides from marine invertebrates. Antimicrob. Agents Chemother. 48(10): 3645-3654.
27. Veeruraj, A., Ravichandran,S., Rameshkuma,G. ( 2008):Antibacterial activity of crab hemolymph on clinical pathogens. J Trend in Applied Sci Res. 3: 174-81.
28.Youqin, K., Liqiao, C., Zhili, D. (2016): Molecular cloning, characterization, and mRNA expression of hemocyanin Subunit in Oriental River Prawn Macrobrachium nipponense. Int J Genomics. 13: 6404817.
29. Zhao, D., Song, S., Wang, Q., Zhang, X., Hu, S., Chen, L. (2009): Discovery of immune-related genes in chinese mitten crab (Eriocheir sinensis) by expressed sequence tag analysis of haemocytes,” Aquaculture. 287(3-4): 297–303.
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