Hemocyte Types Based on Total and Differential Counts in Samia cynthia ricini (Lepidoptera; Saturniidae) Reared on Host Plants Versus an Artificial Diet


Jatuporn Tungjitwitayakul Nujira Tatun


     Hemocytes are an important component of the insect immune system because of their involvement in coagulation, phagocytosis, and encapsulation. The larvae of the eri silkworm, Samia cynthia ricini, has been successfully used as insect model to study the innate immune response and antibacterial activity. This insect grows on host plants, such as cassava and castor leaves, and on artificial diets. Several studies have revealed that artificial diets alter the insect’s immune responses. Hence, it is important to assess the concentration of hemocytes in S. cynthia ricini reared on the host plant (castor leaves) and an artificial diet. The hemocytes of S. cynthia ricini are classified into five types: prohemocytes (PRs), plasmatocytes (PLs), granulocytes (GRs), spherulocytes (SPs), and oenocytoids (OEs). The total hemocyte count (THC) was studied in four developmental stages, including 3rd-, 4th-, and 5th-instar larvae, in addition to pupae. The results indicated that the THC in larvae reared on castor leaves decreased gradually in later developmental stages. In contrast, the THC in larvae reared on an artificial diet were at lower levels in all stages. The differential hemocyte count (DHC) indicated that ratio of each hemocyte type was comparable during larval and pupal stages in which PLs were most abundant, followed by GRs, SPs, PRs, and OEs in both groups. Furthermore, the diet had different effects on the percentage of PLs, GRs, and PRs during larval and pupal stages. Since the immune system of eri silkworm was interfered, this artificial diet may not suitable for rearing system in immunological study aspect.

Keywords: eri-silkworm, hemocytes, castor leave, artificial diet, immunity


Ajamhassani, M., Jalali, S. J., Zibaee, A., Askary, H., & Farsi, M. J. (2013). Immunological responses of Hyphantria cunea (Drury) (Lepidoptera: Arctiidae) to entomopathogenic fungi, Beauveria bassiana (Bals.-Criy) and Isaria farinosae (Holmsk.). Fr. Journal of Plant Protection Research, 53 (2), 110-118.
Andrade, F. G., Negreiro, M. C. C., Gregório, E. A., Moscardi, F., & Falleiros, A. M. F. (2003). Hemocytes of Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae) larvae: morphological and quantitative studies. Acta Microscopica, 12, 59-64.
Bao, Y., Yamano, Y., & Morishima, I. (2005). A novel lebocin-like gene from eri-silkworm, Samia cynthia ricini, that does not encode the antibacterial peptide lebocin. Comparative Biochemistry and Physiology- Part B: Biochemistry and Molecular Biology, 140(1), 127-131.
Bao, Y., Yamano, Y., & Morishima, I. (2007). Induction of hemolin gene expression by bacterial cell wall components in eri-silkworm, Samia cynthia ricini. Comparative Biochemistry and Physiology- Part B: Biochemistry and Molecular Biology, 146(1), 147-151.
Bhagawati, N., & Mahanta, R. (2014). Variation in haemocyte count in haemolymph of different larval stages of eri silkworm on application of malathion: an organophosphorous pesticide. India Journal of Scientific Research and Technology, 2(5), 59-63.
Deka, M., Dutta, S., & Devi, D. (2011). Impact of feeding of Samia cynthia ricini Boisduval (red variety) (Lepidoptera: Saturniidae) in respect of larval growth and spinning. International Journal of Pure and Applied Sciences and Technology, 5(2), 131-140.
Falleiros, A. M. F., Bombonata, M. T. S., & Gregorio, E. A. (2003). Ultrastructural and quantitative studies of hemocytes in the sugarcane borer, Diatraea saccharalis (Lepidoptera: Pyralidae). Brazilian Archives of Biology and Technology, 46, 287-294.
Fukuzawa, M., Tatsuki, S., & Oshikawa, Y. (2004). Rearing of Ostinia palustralis (Lepidoptera: Crambidae) larvae with a switchover of two kinds of artificial diets. Applied Entomology and Zoology, 39(3), 363-366.
Ghasemi, V., Moharramipour, S., & Sendi, J. J. (2014). Impact of pyriproxyfen and methoxyfenozide on hemocytes of the Mediterranean flour moth, Ephestia keuhniella (Lepidoptera: Pyralidae). Journal of Crop Protection, 3(4), 449-458.
Gupta, A.P. (1991). Insect immunocytes and other hemocytes: roles in cellular and humoral immunity, In Gupta, A. P. (Ed.). Immunology of insects and other arthropods (pp. 19-118). CRC Press: Boca Raton.
Han, S. S., Lee, M. H., Kim, W. K., Wago, H., & Yoe, S. M. (1998). Hemocytic differentiation in hemopoietic organ of Bombyx mori larvae. Zoological Science, 15, 371-379.
Hashimoto, K., Yamano, Y., & Morishima, I. (2008a). Induction of tyrosine hydroxylase gene expression by bacteria in the fat body of eri-silkworm, Samia cynthia ricini. Comparative Biochemistry and Physiology- Part B: Biochemistry and Molecular Biology, 149(3), 501-506.
Hashimoto, K., Yamano, Y., & Morishima, I. (2008b). Cloning and expression of a gene encoding gallerimycin, a cysteinerich antifungal peptide, from eri-silkworm, Samia cynthia ricini. Comparative Biochemistry and Physiology- Part B: Biochemistry and Molecular Biology, 150(2), 229-232.
Hirayama, C., Konno, K., Wasano, N., & Nakamura, M. (2007). Differential effects of sugar-mimic alkaloids in mulberry latex on sugar metabolism and disaccharidases of Eri and domesticated silkworms: Enzymatic adaptation of Bombyx mori to mulberry defense. Insect Biochemistry and Molecular Biology, 37, 1348-1358.
Huang, F., Yang, Y. Y., Shi, M., Li, J. Y., Chen, Z. Q., Chen, F. S., & Chen, X. X. (2010). Ultrastructural and functional characterization of circulating hemocytes from Plutella xylostella larva: Cell types and their role in phagocytosis. Tissue and Cell, 42(6), 360-364.
Jalali, J., & Salehi, R. (2008). The hemocyte types, differential and total count in Papilio demoleus L. (Lepidoptera: Papilionidae) during post-embryonic development. Munis Entomology& Zoology, 3(1), 199-216.
Kataoka, K., & Imai, T. (1986). Cocoon quality and physiological properties of the cocoon filament produced by silkworm reared on mulberry leaves and on artificial diet. The Journal of Sericultural Science of Japan, 55(2), 112.
Khosravi, R., Jalali, S., & Ghasemi, V. (2012). Identification of hemocytes in carob moth, Ectomoyelois ceratoniae Zeller (Lepidoptera: Pyralidae) larvae. Journal of Plant Pests Research, 2, 29-39.
Kishimoto, K., Fujimoto, S., Matsumoto, K., Yamano, Y., & Morishima, I. (2002). Protein purification, cDNA cloning and gene expression of attacin, an antibacterial protein, from eri-silkworm, Samia cynthia ricini. Insect Biochemistry and Molecular Biology, 32(8), 881-887.
Konno, K., Ono, H., Nakamura, M., Tateishi, K., Hirayama, C., Tamura, Y., … Kohno, K. (2006). Mulberry latex rich in anti-diabetic sugar-mimic alkaloids forces dieting on caterpillars. Proceedings of National Academy of Sciences of the United States of America, 103, 1337-1341.
Kurihara, Y., Shimazu, T., & Wago, H. (1992). Classification of hemocytes in the common cutworm Spodoptera litura (Lepidoptera: Noctuidae) II. Possible roles of granular plasmatocytes and oenocytoids in the cellular defense reactions. Applied Entomology and Zoology, 27, 237-242.
Lavine, M. D., & Strand, M. R. (2002). Insect haemocytes and their role in immunity. Insect Biochemistry Molecular and Biology, 32, 1295-1309.
Ling, E., & Yu, X. Q. (2006). Hemocytes from the tobacco hornworm Manduca sexta have distinct functions in phagocytosis of foreign particles and self dead cells. Developmental and Comparative Immunology, 30, 301-309.
Mahalingam, V., & Muralirangan, M. C. (1998). Age and sex correlated haemocytic profile of Atractomorpha crenulata (Fabricius) (Insecta: Orthoptera: Pyrgomorphidae) in laboratory cultures reared on Ricinus communis. Journal of Orthoptera Research, 7, 29-32.
Onoe, H., Matsumoto, A., Hashimoto, K., Yamano, Y., & Morishima, I. (2007). Peptidoglycan recognition protein (PGRP) from eri-silkworm, Samia cynthia ricini; protein purification and induction of the gene expression. Comparative Biochemistry and Physiology- Part B: Biochemistry and Molecular Biology, 147(3), 512-519.
Pletcher, S. D., Macdonald, S. J., Marguerie, R., Certa, U., Stearns, S. C, Goldstein, D. B., & Patridge, L. (2002). Genome-wide transcript profiles in aging and calorically restricted Drosophila melanogaster. Current Biology, 12, 712-723.
Ponton, F., Wilson, K., Cotter, S. C., Raubenheimer, D., & Simpson, S. J. (2011). Nutritional immunology: a multi-dimensional approach. PLoS Pathogens, 7(12), e1002223.
Ribeiro, C., & Brehélin, M. (2006). Insect haemocytes: What type of cell is that? Journal of Insect Physiology, 52, 417-429.
Richards, E. H., & Edwards, J. P. (1999). Parasitization of Lacanobia oleracea (Lepidoptera: Noctuidae) by the ectoparasitic wasp Eulophus pennicornis: effects of parasitization, venom and starvation on host haemocytes. Journal of Insect Physiology, 45, 1073-1083.
Ruiz, E., López, M. C., Rivas, F. A., Sánchez, A. Y., & Moncada, L. I. (2015). Comparison of hemocytes of V-instar nymphs of Rhodnius prolixus (Stål) and Rhodnius robustus (Larousse 1927), before and after molting. Revista de la Faculted de Medicina, 63(1), 11-17.
Russo, J., Brehelin, M., & Carton, Y. (2001). Hemocyte changes in resistant and susceptible strains of D. melanogaster caused by virulent and avirulent strains of the parasitic wasp Leptopilina bouladi. Journal of Insect Physiology, 47, 167-172.
Schmitz, A., Anselme, C., Ravallec, M., Rebuf, C., Simon, J. C., Gatti, J. L., & Poirié, M. (2012). The cellular immune response of the Pea Aphid to foreign intrusion and symbiotic challenge. PLoS One, 7, e42114.
Siva-Jothy, M. T., & Thompson, J. J. W. (2002). Short-term nutrient deprivation affects immune function. Physiological Entomology, 27, 206-212.
Shapiro, M. (1979). Changes in hemocyte populations, In Gupta, A.P. (Ed.). Insect hemocytes, development, forms, functions and techniques. New York: Cambridge university press.
Strand, M. R., & Pech, L. L. (1995). Immunological basis for compatibility in parasitoid-host relationships. Annual Review of Entomology, 40, 31-56.
Suzuki, Y., Kawanishi, S., Yamazaki, T., Aoki, A., Saito, H., & Asakura, T. (2015). Structural determination of the tandem repeat motif in Samia cynthia ricini liquid silk by solution NMR. Macromolecules, 48, 6574-6579.
Takahashi, S., & Enomoto, G. (1995). The initial phase of encapsulation of silicone oil injected in Samia cynthia ricini (Lepidoptera, Saturniidae): The innermost structure of the developing capsule. Zoological Science, 12(3), 303-309.
Tan, J., Xu, M., Zhang, K., Wang, X., Chen, S., Li, T., … Cui, H. (2013). Characterization of hemocytes proliferation in larval silkworm, Bombyx mori. Journal of Insect Physiology, 59, 595-603.
Tojo, S., Naganuma, F., Arakawa, K., & Yokoo, S. (2000). Involvement of both granular cells and plasmatocytes in phagocytic reactions in the greater wax moth, Galleria mellonella. Journal of Insect Physiology, 46, 1129-1135.
Tungjitwitayakul, J., & Tatun, N. (2017). Comparison of biological and biochemical parameters of eri-silkworms, Samia cynthia ricini (Lepidoptera: Saturniidae), reared on artificial and natural diets. Journal of Entomology and Zoology Studies, 5(2), 314-319.
Vogelweith, F., Moret, Y., Monceau, K., Thiéry, D., & Moreau, J. (2016). The relative abundance of hemocyte types in polyphagous moth larva depends on diet. Journal of Insect Physiology, 88, 33-39.
Yamashita, M., & Iwabuchi, K., (2001). Bombyx mori prohemocyte division and differentiation in individual microcultures. Journal of Insect Physiology, 47, 325-331.
Yang, S. Y., Ruuhola, T., Haviola, S., & Rantala, M. J. (2008). Effects of host-plant shift on immune and other key life-history traits of an eruptive Geometrid, Epirrita autumnata (Borkhausen). Ecological Entomology, 33, 510-516.
Zhou, Z. H., Yang, H. J., Chen, M., Lou, C. F., Zhang, Y. Z., Chen, K. P., … Zhong, B. X. (2008). Comparative proteomic analysis between the domesticated silkworm (Bombyx mori) reared on Fresh mulberry leaves and on artificial diet. Journal of Proteome Research, 7, 5103-5111.

Research Articles


How to Cite
TUNGJITWITAYAKUL, Jatuporn; TATUN, Nujira. Hemocyte Types Based on Total and Differential Counts in Samia cynthia ricini (Lepidoptera; Saturniidae) Reared on Host Plants Versus an Artificial Diet. Naresuan University Journal: Science and Technology (NUJST), [S.l.], v. 27, n. 3, p. 82-94, july 2019. ISSN 2539-553X. Available at: <https://www.journal.nu.ac.th/NUJST/article/view/Vol-27-No-3-2019-82-94>. Date accessed: 26 feb. 2024. doi: https://doi.org/10.14456/nujst.2019.28.