Greener Journal of Biological Sciences Vol. 7 (3), pp. 025-033, May 2017.
© 2017 Authors
Manuscript Number: 032017040
Qualitative and Quantitative Changes of Haemocytes from Honeybee, Apis mellifera (L.) (Hymenoptera:
Apidae) After Intrahaemocoelic Injection with Bacillus thuringiensis
Emad M.S. Barakat*1, Walaa A. Moselhy2, Zeinab A. Shouaib3, Mostafa A. Taha4
1Department of Entomology, Faculty of Science, Ain Shams University, Abbassia, Cairo, Egypt.
2, 3, 4Department of zoology, Faculty of Science, Al-Azhar University, Naser city, Cairo, Egypt.
The present work was carried out aiming to characterize the haemocytes from different developmental stages of honeybee, Apis mellifera (L.) after experimental injection of Bacillus thuringiensis (B.t.) into the haemocoel. Six haemocyte types were recognized in pupae: prohaemocytes (PRs), granulocytes (GRs), oenocytoides (OEs), spindle-shaped cells (SPLs), plasmatocytes (PLs), and adipohaemocytes (ADs). Eight types were found in larvae; in addition to the previous mentioned haemocyte types, spherulocytes (SPs) and eosinophil cells (EOs), while ten types were recognized in the adult stage; the previous mentioned eight types plus micronucleocytes (MIs) and macronucleocytes (MAs). Due to bacterial injection, certain pathological consequences were observed in the defined haemocytes such as vacuolization in the cytoplasm, distortion of the cell membrane and pycnosis in the nuclei. Moreover, the differential and total haemocyte counts were found to increased or decreased depending on the developmental stage and the time of haemolymph collection after bacterial injection. Other observed cellular responses against bacterial injection were phagocytosis and nodule formation.
Key words: Apis mellifera, Bacillus thuringiensis, haemolymph, haemocytes, THC, DHC, cellular immunity, phagocytosis, nodule formation.
Abdel-Rahman, M. F. 2014. Role of pollen and/or bee bread avail ability in brood rearing and cellular immune system of honey bees. J. Plant Prot. and Path., Mansoura Univ., Vol. 5 (12): 1125 – 1137.
Abrol, D.P. (1996). Effect of mite parasitosis on blood counts of honey bees Apis mellifera L. and Apis cerana indica F. (Hymenoptera : Apidae).Insect Environ., 2(1):18-19.
Agripina Sapcaliu, I. Radoi, Crengula Pavel, N. tudor, Eliza Caula, A. siceanu, F. meiu 2009. Research Regarding haemocte profile from Apis mellifera carpatica bee haemolymph originated in the south of Romania Medicina veterinara VOL. XLII.
AlFonso, T.B. and Jones, B.W. 2002. Gcm2 promotes glial cells differentiation and is required with glial cells missing for macrophage development in Drosophila. Devel. Biol. 248, 369-383.
Anita Giglio; Piero Giulio; Giulianini 2013. Phenoloxidase activity among developmental stages and pupal cell types of the ground beetle Carabus (Chaetocarabus) lefebvrei (Coleoptera, Carabidae) Department of Biology, Ecology and Earth Sciences, University of Calabria.
Arnold, J.W. and Hinks, C.F. 1979. Insect haemocytes under light microscopy: technique. In: Insect Haemocytes (A.P. Gupta, ed.). Cambridge Univ. Press, Cambridge.
Ashida, M., Ochiai, M. and Niki, T. 1988. Immunolocalization of prophenoloxidase among hemocytes of the silkworm, Bombyx mori. Tissue & Cell 20, 599-610.
Aubert, A., Richard, F.-J. 2008. Social management of LPS-induced inflammation in Formica polyctena ants. Brain, Behavior, and Immunity 22: 833–837.
Bailey and Ball (1991): Honey Bee Pathology 2nd edn. Academic Press, London.
Barakat, E.M.S. 2000. Haemocytic changes in honey bee, Apis mellifera (L.) following injection of bacteria. Ain Shams Sci. Bull., 38: 500-517.
Barakat, E.M.S. 2001. The cellular defence reactions of the honey bee, Apis mellifera in response to injected
bacterium, Pseudomonas aeruginosa. Ain Shams Sci. Bull., 39: 214-227.
Barakat, E.M.S.; AboKersh, M.O. Gomaa, S.A. 2016. Haemocyte Activity and Cellular Defense Reactions in Various Larval Instars of Honey Bee (Apis mellifera L.) following Natural and Experimental Bacterial Infections, Journal of Biological Sciences Vol. 6 (2), pp. 020-033.
Barakat, E.M.S.; Meshrif, W.S. and Shehata, M.G. 2002. Changes in the haemolymph of the desert locust, Schistocerca gregaria after injection Bacillus thuringiensis. J. Egypt. Acad. Soc. Environ. Develop. (A.Entomology), 2: 95-115.
Bardoloi, S. and Hazarika, L. K. 1992. Seasonal variation of body weight, lipid reserves, blood volumes, and
haemocyte population of Antheraea assama (Lepidoptera: Saturniidae). Environ. Entomol., 21(6): 1398-1403.
Bedick, J.C.; Tunaz, H.; Aliza, A.R.N.; Putnam, S.M.; Ellis, M.D. and Stanley, D.W. 2001. Eicosanoids act in nodulation reactions to bacterial infections in newly emerged adult honey bees, Apis mellifera, but not in older foragers. Comp. Biochem. Physiol., 130:107–117.
Bozena and Andrzej 2003. The influence of different diets on haemocytes of adult worker honey bees, Apis mellifera Apidologie 34 (2003) 97–102.
Brehelin, M. and Zachary, D. 1986. Insect haemocytes: a new classification to rule out the controversy. In: "Immunity invertebrates, cells, molecules and defense reactions" (Brehélin, M., ed.). Heidelberg: Spring Verlag. pp. 37-48.
Evans, J.D.; Aronstein, K. Chen, Y.P.; Hetru, C.; Imler, J-L.; Jiang, M.; Kanst, M.; Thompson, G.J.; Zou, Z. and Hultmark, D. 2006. The Immune pathways and defence mechanisms in honey bees Apis mellifera. Inse. Mol. Biol., 15(5): 645–656.
Faye, I. (1978): Insect immunity: early fate of bacteria injected in Saturniid pupae. J. Invertebr. Pathol., 31: 19-26.
Gilliam, M. and Shimanuki, H. 1970. Total hemocyte counts of honey bee larvae (Apis mellifera L.) from various elevations. J. Cellul. and Molec. Life Scienc. (CMLS), 26 (9).
Gilliam, M. 1973. Age-dependent variation of differential haemocyte counts of developing worker honeybees. J. Apic. Res., 12 (1): 61-64.
Glinski, Z. and Jarosz, J. 1995. Mechanical and biochemical defenses of honeybees. Bee World, 76(1): 110-118.
Glinski, Z. and K. Grazgorczyk, 1995. Cellular defense reactions in the honey bee in environment non-polluted with
heavy metals. Annales- Universitatis Mariae -Curie Skodwska -Sectio dd; Medicina Veterinaria, 50(13): 131-137.
Glinski, Z. and Klimont 1987a. Effect of Varroa jacobsoni infestation on the blood cells of worker bees. Medycyna Weteryunaryjna, 43(9): 546-549; (Review of Applied Entomology, 6730/88).
Glinski, Z. and Klimont 1987b. Activity of haemocytes of worker honeybee during nature infestation with Varroa jacobsoni. Medycyna Weteryunaryjna, 43(11): 664-667; (Apic. Abst., 251/90).
Godfrey, S.S., Bull, C.M., Murray, K., Gardner, M.G. 2006. Transmission mode and distribution of parasites among groups of the social lizard Egernia stokesii. Parasitol. Res., 99: 223–230. Gregore, A. and Bowen, I.D. 1998. Histopathological and histochemical changes in honey bee larvae (Apis mellifera) after infection with Bacillus larvae, the causative agent American foulbrood disease. Cell-Biol.- Internatio., 22(2): 137-144.
Gunnarsson, S.G. and Lackie, A.M. 1985. Haemocytic aggregation in Schistocerca gregaria and Periplaneta americana as a response to injected substances of microbial origin. J. Invertebr. Pathol., 46: 312-319.
Gupta, A.P., (1979): Insect haemocytes. Cambridge University Press, Cambridge. pp. 614.
Heike Gätschenberger, Olaf Gimple, Jürgen Tautz and Hildburg Beier 2011. Honey bee drones maintain humoral immune competence throughout all life stages in the absence of vitellogenin production BEEgroup, Biocentre, University of Würzburg, Am Hubland, D-97074 Würzburg, Germany.
Jones, J.C., 1962. Current concepts concerning insect haemocytes. Amer. Zool. 2, 209-246.
Kanost, M.R., H. Jiang, and X.Q. Yu, 2004. Innate immune responses of a lepidopteran insect, Manduca sexta. Immunol. Rev. 198, 97-105.
Lavine M.D. and Strand, M.R. 2002. Insect haemocytes and their role in immunity. Insect Biochem. Molec. Biol. 32, 1295-1309.
Liu F., Xu, Q., Zhang, Q., Lu, A., Beerntsen, B.T. and Ling, E. 2013. Hemocytes and hematopoiesis in the silkworm, Bombyx mori. I.S.J. 10, 102-109.
Maragarita M. lops Maragarita ez-Uripe, Warren B. Sconiers, Steven D. Frank and David R. Tarpy 2016. Reduced cellular immune response in social insects Dept. Entomology & Dept. of Applied Ecology.
Maria J. Kirrane, lilial de Guzaman, B.t.h Holloway Amand M. Frake, Thomas Rinder & Padriag M. 2015. Phenotypic & Genetic analysis of the varroa sensitive hygienic trait in Russian honeybee (Hymenoptera: Apidae) colonies.
Momen, S. A., Salem, D. A. M., Barakat, E. M. S. and salama, M. S. 2012. The role of prophenoloxidase Activation System IN Cellular Defense Mechanisms in the Haemolymph of the Desert Locust, Schistocerca gregaria (Forskal).M.Sc. Thesis, Sci. Facu., Ain Shams Univ.
Mori, H. 1979. Embryonic haemocytes. In: Origin and development in insects haemocytes. (Eds., Gupta, A.P.). pp. 3-27. Cambridge, Camridge university press.
Neuwrith, M., 1973. The structure of the haemocytes of Galleria mellonellaLepidoptera). J Morph. 139, 105-124.
Nour, M.N., 1998. Chronic bee paralysis virus(CPV) in honey bee colonies in Egypt. J. Agric. Sci. Mansoura Univ.,
Ottaviani, E. (2005). Insect immunorecognition. ISJ 2: 142-151. ISSN 1824-307X.
Papadopoulou-Karabela, K.; Lliads, N. and Liakos, V. 1993. Haemocyte changes in honeybee (Apis mellifer L.) artificially infected by Pseudomonas aeruginosa. Apidol., 24: 81-82.
Ratcliffe, N. A. and Rowley, A. F. 1979. Role of haemocytes in defense against biological agents. In Insect haemocytes: Development, Forms, Functions and Techniques (A.P> Gupta,) 331-414. London: Cambridge Univ. Press (GR& PLS phagocytosis).
Rowley, A.F. and Ratcliffe, N.A. 1980. Insect erythrocyte agglutinines. In vitro, opsonization experiments with Clitummus extradentatus and Periplaneta americana haemocytes. Immunol., 40: 483-492.
Salt, G. (1970). The cellular defense reactions of insects. Cambridge Monographs in Experimental Biology. No 16, pp. 118, Cambridge Univ. Press. London and New york.
Sanjayan K.P., Ravikumar, T. and Albert, S. 1996. Changes in the haemocyte profile of Spilostetethus hospes (Fab) (Heteroptera: Lygaeidae) in relation to eclosion, sex and mating. J. Biosci. 21(6), 781-788.
Sarag El-Dien, F.S. (1999): Studies on the varroatosis "Varroa jacobsoni Oud." Ph. D. Thesis, Agric. Fac., of Tanta Univ., Egypt.
Sawsan S. El Mohandes; Emad A. Nafea and Asmaa M. Fawzy 2010. Effect of different feeding diets on the haemolymph of the newly emerged honeybee workers Apis mellifera L. Acad. J. biolog. Sci., 3 (1): 213 – 22.
Schmidt R. Martin, Axel Brockmann, Christian W.W. Pirk, David W. Stanleyc and Jürgen Tautz 2008: Adult honeybees (Apis mellifera L.) abandon hemocytic, but not phenoloxidase-based immunity. Journal of Insect Physiology 54(2):439-44 ·
Shapiro, M. 1979. Changes in the haemocyte populations In: Insect Haemocytes (Gupta AP, ed) Cambridge Univ. Press, Cambridge.
Siddiqui M.I. and Al-Khalifa, M.S. 2012. Circulating haemocytes in insects: phylogenic review of their types. Pakistan J. Zool. 44(6), 1743-1750.
Sorescu, I. and Dragomir, C. 2003. Differential haemocyte count and the phagocytic index in experimental bacterial infections and in field ascosphaerosis of honey bee (Apis mellifera L.) at various ontogenetic stages. Studies-and-Researchers-in-Veterinary-Medicine, 9: 133-168.
Strand, M.R., 2008. The insect cellular immune response. Insect Sci. 15, 01-14.
Tauber, O.E. and Yeager, J.F. 1935. On the total blood counts of insects. I. Orthoptera, Odonata, Hemiptera and Homoptera. Ann. Entomol. Soc. Amer., 28: 229-240.
Tojo, S.; Naganuma, F.; Arakawa, K. and Yokoo S. 2000. Involvement of both granular cells and plasmatocytes in phagocytic reactions in the greater wax moth, Galleria mellonella. J. Insect Physiol., 46: 1129–1135.
Van-Steenkiste, D. 1988. The haemocytes of the honeybee: typology, blood spectrum and cellular defence reactions. Bdo.proefschrift, Doctor in de wetenschappen, Rijksuniversiteit Gent, Belgium, (Apic. Abst 1173/89).
Wienands, A.; Strich, H. and Madel, G. 1987. Haemocytic and bacteriological aspects of the haemolymph of Apis mellifera L. (Hymenoptera, Apidae). Zentrlblatt-fur-Bakteriologie,-Mikrobilogie-und-Hygiene,-A. pp. 265, 489; Ba (Apic. Abst. 1264/89).
Wille, H. and Veechi, M.A. 1974. Untersuchungen Uber die haemolymphe der honigbien (Apis mellifera L.) 5. Morphologie der Leukozyten und vier krankheits elementen. Mitt Schweiz Entomol. Ges., 47: 133-149.
Zakaria M.E. 2007. The Cellular Immunity Responses in The Haemolymph Of Honey Bee Workers Infected By American Foulbrood Disease (AFB), Journal of Applied Sciences Research, 3(1): 56-63.
Zakaria, M.E. 1997. Current concepts concerning, haemocytes of honey bees and their features according to varroa parasitism. M. Sc. Thesis, Agric. Fac., Cairo Univ. pp: 172.
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