Study of total hemocyte count and total plasma protein in immune system of freshwater Crayfish (Astacus leptodactylus)

Samnejhad, Arash; Afsharnasab, Mohammad

doi:10.22113/jmst.2016.8236

Study of total hemocyte count and total plasma protein in immune system of freshwater Crayfish (Astacus leptodactylus)

Document Type : Original Manuscript

Authors

arash samnejhad ¹

mohammad afsharnasab ²

¹ Iranian Artemia Research Center, Orumia, Iran

² Iranian Fisheries Research organization, Karajh, Iran

10.22113/jmst.2016.8236

Abstract

Abstract In this present study, hemocytes defence cells and total plasma protein had been studied in immune system of freshwater Crayfish Astacus leptodactylus. About one-hundered fresdhwater Crayfish (A.leptodactylus) with average weight of 25-40g were purchased from aras damreservours in west Azarbayjan province and transported to Iranian artemia research center of orumya province (25 pieces of Crayfish were considered surplus for possible losses). Before experimernt the Crayfish were acclimated for two days in the laboratory and then disinfected by oxytetracycline antibiotic and finally, an equal number (15 pieces of Crayfish in each glass aquariums) transported to 5 glass aquariums. In order to study hemocyte and plasma total protein, the hemolymph samples were withdrawn from abdominal second segments of Crayfish in interval hours (0, 6, 12, 24, 48, 96, 144, 240 and 336). Result showed that the mean of hemocytes was 27-114 CFU ml-1, and the highest and lowest value of hemocytes were 180 and 12 CFU ml-1 in 12 and 336 hours after experiment times, respectively. Also based on the results, the semigranular cells(SGC) and granular cells(GC) were approximately comprised about 58-65% and 28-37%, respectively and hyaline cells(HC) about was 3-6%. Based on total plasma protein results, the mean of total plasma protein was 1-2 Gr dl-1, and the highest and lowest value of total plasma protein were 2/8 and 0/5 Gr dl-1 in 12 and 336 hours after experiment times.

Keywords

Hemocytes

Granular cells

Semigranular cells

Hyaline cells

Total plasma protein

20.1001.1.20088965.1395.15.1.5.0

Subjects

سامکوکیائی، آ. 1390. بررسی تجربی بیماریزائی باکتری آئروموناس هیدروفیلا در شاهمیگوی آب شیرین (Astacus leptodactylus) با تا کید برتاثیر دما در روند بیماریزائی باکتری. پایان نامه دکتری تخصصی، گروه بهداشت و بیماریهای آبزیان، دانشکده تخصصی دامپزشکی، دانشگاه علوم و تحقیقات، 101 صفحه.

Battison, A., Cawthorn, R. and Horney, B. 2003.Classification of Homarus americanus hemocytes and the use of differential hemocyte counts in lobster infected with Aerococcus viridians Var. homari (Gaffkemia). Invert. pathol. 84: 177-197.

Coeurdacier, J.L., Dutto, G., Gasset, E. and blancheton, J.P. 2011.Is total serum protein a good indicator for welfare in reared sea bass (Dicentrarchus labrax)?. Aqua. Liv. Re. 24: 121-127.

Dawson, R. D. and Bortolotti, G.R. 1997.Total plasma protein level as an indicator of condition in wild American kestrels (Falco sparverius). J. Zool. 75: 680–686.

De le Court, C., Aguilera, E. and Recio, F. 1995.Plasma chemistry values of free-living white spoonbills (Platalea leucorodia). Com. Biochem Physiol. 112A: 137–141.

Gargioni, R. and Barracco, M.A. 1998. Hemocytes of the palaemonids Macrobrachium rosenbergii and M. acanthurus, and of the penaeid Penaeus paulensis. J. Morphol. 236: 209-221.

Gillespie, J.p., Kanost, M.R. and Trenczek, T. 1997.Biological mediators of insect immunity. Annual. Rev. Entomol. 42: 611-643.

Giulianini, P.G., Bierti, M., Lorenzon, S., Battistella, S. and Ferrero, E.A. 2007.Ultra structural and functional characterization of circulating hemocytes from the freshwater Crayfish Astacus leptodactylus: Cell types and their role after invivo artificial non-self challenge. Micron. 38: 49-57.

Gross, P.S., Bartle, T.C., Browdy, C.L., Chapman, R.W., Warr, G.W. 2001.Immunegene discovery by expressed sequence tag analysis of hemocytes and hepatopancreas in the pacific white shrimp, Litopenaeus vannamei, and the Atlantic white shrimp, L. setiferus. Dev. Com. Immunol. 25:565-577.

Holdich, D.M., Harlioglu, M.M. and Firkins, I. 1997.Salinity adaptations of crayfish in British waters with particular refrence to Austropotamobius pallipes, Astacus leptodactylus and Pasifastacus leniusculus, Estuarine, Coast. Shelf. sci. 44:147-154.

Hose, J.H., Martin, G.G. and Gerard, A.S. 1990.Adecapod hemocyte classification scheme integrating morphology, cytochemistry, and function. Biol. Bullet. 178: 33-45.

Jiravanichpaisal, P., Lee, B.L. and Soderhall, K. 2006a.cell – mediated immunity in arthropods: Hematopoiesis, coagulation, melanization and opsonization. Immunol. 211:213-236.

Johansson, M.W., Keyser, P., Sritunyalucksana, K. and Soderhall, K. 2000.Crustacean hemocytes and haematopoiesis. Aqua. 191:45-52.

Jussila, J., Jago, J., Tsvetnenko, E., Dunstan, B. and Evans, L.H. 1997.Total and differential hemocyte counts in western rock lobsters (Panulirus Cygnus George) under post-harvest stress. Mar. Fwater. Re. 48: 863-867.

Kakoolaki, S., Sharifpour, I., Soltani, M., Mousavi, H.A.E., Mirzargar, S., and Rostami, M. 2010.Selected morpho-chemical features of hemocytes in farmed shrimp. Fenneropenaeus indicus in iran. I. J Fish. Sci. 9(2):219-232.

Liu, H. 2008. Functional studies of some immune relevant genes in a crustacean. Acta universitatis upsaliensis. Digital comprehensive summaries of Uppsala Dissert. Facul. Sci. technol. 435: 61p.

Lin, X. 2010. Hematopoiesis in a crustacean. Acta universitatis Upsaliensis. Digital comprehensive summaries of Uppsala Dissert Faculty. Sci. Technol. 723:46pp.

Lorenzon, S., Francese, M., Smith, V.J., and Ferrero, E.A. 2001.Heavey metals affect the Circulating hemocyte number in the shrimp Palaemon elegans. Fish. Shell. Immunol. 11: 459-472.

Matinfar, A. 2007.Prancipal program of Freshwater Crayfish Astacus leptodactylus. Jahad agriculture organization. I. Fish. Re. Organ. 10:118p.

Mix, M.C. and Sparks, A.K. 1980. Hemocyte classification and differential counts in Dungeness Crab Cancer magister. J. Invert. Pathol. 35(2): 134-143.

Mohajeri, J., Afsharnasab, M., Jalali, B., Kakoolaki, S., Sharifrohani, M., and Haghighi, A. 2011.Immunological and Histopathological changes in Penaeus semisulcatus challenged with Vibrio harveyi. I. J. Fish. Sci. 10(2): 254-265.

Ozbay, G. and Riley, G.J. 2002.An analysis of refractometry as a method of determining blood total protein concentration in the American lobster Homarus americanus (Milne Edwards). Aqua. Re. 33: 557-562.

Riche, M. 2007.Analysis of refractometry for determining total plasma protein in hybrid striped bass (Morone chrysops & Morone saxatilis) at various salinities. Aqua. 264: 279−284.

Rodriguez, J. and Le Moullac, G. 2000.State of the are of immunological tools and health control of penaeid shrimp. Aqua. 191:109-119.

Samcookiyaei, A., Afsharnasab, M., Razavilar, V., Motalebi, A., Asadpor, Y., Yahyazade, M.Y. and Nekouifard, A. 2012. Experimentally pathogenesis of Aeromonas hydrophila in freshwater Crayfish (Astacus leptodactylus). I. J. Fish. Sci. 11(3): 644-656.

Schoech, S. T. and Bowman, R. 2003.Does differential access to protein influence differences in timing of breeding of Florida scrub-jays (Aphelocoma coerulescens) in suburban and wildland habitats? Auk. 120: 1114–1127.

Smith, V.J. 1991.Invertebrate immunology: phylogenetic, ecotoxicological and biomedial implications. Com. Haematol. Int. 1: 61-76.

Soderhall, K. and Smith, V.J. 1983.Separation of the haemocyte populations of Carcinus meanas and other marine decapods, and prophenoloxidase distribution. Dev. Com. Immunol. 7:229-239.

Soderhall, K., Wingren, A., Johansson, M.W. and bertheussen, K. 1985.The cytotoxic reaction on of hemocytes from the freshwater crayfish Astacus astacus. Cell. Immunol. 94(2): 326-332.

Soderhall, K. and Cerenius, L. 1998.Role of the prophenoloxidase-activating system in invertebrate immunity. Curr. Opinion. Immunol, 10: 23-28.

Sritunyalucksana, K. and Soderhall, K. 2000.The proPo and cloting system in crustaceans. Aqua. 191: 53-69.