Taze dut yapraklarında yetiştirilen koza ören ve koza öremeyen ipekböceği larvalarının Bombyx mori (L.,1758) (Lepidoptera: Bombycidae) hemolenfinde biyokimyasal değişimler: koza örememe sendromunun olası nedenleri

Öz

Bombyx mori (L., 1758) (Lepidoptera: Bombycidae)’de görülen koza örememe sendromu, ipekböcekçiliği sektöründe ciddi bir sorun olarak görülmektedir. Üre metabolizmasının koza örme başlangıcında önemli olması, bu metabolizmanın koza örememe sendromunda olası rolünün incelenmesi gereğini doğurmuştur. Bu çalışmanın amacı, ipekböceği hemolenfinde ve dut yapraklarında üreaz aktivitesi ve L-arginin konsantrasyonlarını saptayarak, koza örememe sendromunda üre metabolizmasının rolünü araştırmaktır. Bu parametreler ek olarak hemolenf örneklerinde üre konsantrasyonları da ölçülmüştür. Koza ören grubun hemolenfinde üreaz aktivitesi (25 ± 5,8 units/l), koza örmeyen gruba (10,9 ± 2,4 units/l) göre iki kat daha yüksek bulunmuştur (P < 0.05). Koza ören larvaların hemolenfinde üre konsantrasyonu 5. dönemin 5. gününden (137 ± 13 mg/l) 7. gününe önemli seviyede azalırken (97 ± 17 mg/l; P < 0.01), koza öremeyen larvalarda hemen hemen sabit kalmıştır (149 ± 19 mg/l’den 166 ± 4 mg/l’ye). Koza öremeyen larvaların hemolenf örneklerinde L-arginin konsantrasyonu (4.55 ± 0.48 mM), koza ören larva grubundan anlamlı ölçüde yüksek bulunmuştur (2.72 ± 0.45 mM; P < 0.01). Hemolenfteki üreaz aktivitesi ve L-arginin konsantrasyonlarında gözlemlenen değişiklikler benzer şekilde dut yapraklarında da saptanmıştır. Bu sonuçlar, üre metabolizmasındaki olası bir baskılanmanın ipekböceklerinde görülen koza örememe sendromuna katkıda bulunabileceği ve/veya bunun sonucu olabileceğini düşündürmektedir.

Anahtar Kelimeler

• L-arginin
• koza örememe sendromu
• ipekböceği
• üre
• üreaz

Biochemical changes in hemolymph of spinning and non-spinning silkworm larvae, Bombyx mori (L., 1758) (Lepidoptera: Bombycidae), reared on fresh mulberry leaves: possible reasons for non-spinning syndrome

Abstract

Non-spinning syndrome in Bombyx mori (L., 1758) (Lepidoptera: Bombycidae) is a serious issue for the sericulture industry. Determination of urea metabolism as an important parameter at the onset of spinning has shown the need for examining the role of urea metabolism in the non-spinning syndrome. The aim of this study was to investigate role of urea metabolism in the non-spinning syndrome by evaluating urease activity and L-arginine concentrations in the silkworm hemolymph and mulberry leaves. Additionally, urea concentrations were determined in hemolymph samples. Urease activities in hemolymph samples were almost twice as high in spinning larvae (SL) than in non-spinning larvae, 25 ± 5.8 vs 10.9 ± 2.4 units/l (P < 0.05). Urea concentrations in the SL hemolymph decreased significantly from day 5 (137 ± 13 mg/l) to day 7 (97 ± 17 mg/l) of the fifth instar (P < 0.01), it remained almost constant in NSL hemolymph (149 ± 19 to 167 ± 4 mg/l). Additionally, L-arginine concentrations in hemolymph samples obtained from NSL of 4.55 ± 0.48 mM were significantly higher than in SL at 2.72 ± 0.45 mM (P < 0.01). Changes in urease activity and L-arginine concentrations in hemolymph were similarly observed in mulberry leaves. These results suggested that changes in urea metabolism may cause or contribute to non-spinning syndrome in silkworms.

Keywords

• L- arginine
• non-spinning syndrome
• silkworm
• urea
• urease

References

1. Chakrabarty, S. & B. B. Kaliwal, 2012. Application of arginine, histidine and their mixture on economic traits of the silkworm, Bombyx mori L. DAV International Journal of Science, 1 (2): 107-111.
2. Cochran, D. G., 1985. “Nitrogen Excretion, 467-576”. In: Comprehensive Insect Physiology Biochemistry and Pharmacology (Eds. G. A. Kerkut & L. I. Gilbert). Pergamon Press, Oxford, 891pp.
3. Dong, H. L., S. X. Zhang, H. Tao, Z. H. Chen, X. Li, J.F. Qiu, W. Z. Cui, Y. H. Sima, W. Z. Cui & S. Q. Xu, 2017. Metabolomics differences between silkworms (Bombyx mori) reared on fresh mulberry (Morus) leaves or artificial diets. Scientific Reports, 7 (1): 10972.
4. Etebari, K., A. R. Bizhannia, R. Sorati & L. Matindoost, 2007. Biochemical changes in haemolymph of silkworm larvae due to pyriproxyfen residue. Pesticide Biochemistry and Physiology, 88 (1): 14-19.
5. Hirayama, C., K. Konno & H. Shinbo, 1997. The pathway of ammonia assimilation in the silkworm Bombyx mori. Journal of Insect Physiology, 43 (10): 959-964.
6. Hirayama, C., M. Sugimura, H. Saito & M. Nakamura, 2000a. Purification and properties of urease from the leaf of mulberry, Morus alba. Phytochemistry, 53 (3): 325-330.
7. Hirayama, C., M. Sugimura, H. Saito & M. Nakamura, 2000b. Host plant urease in the hemolymph of the silkworm, Bombyx mori. Journal of Insect Physiology, 46 (10): 1415-1421.
8. Jyothi, M., M. Pratap & N. S. Thimma, 2014. Studies on biochemical constituents of different genotypes of Morus alba L. International Journal of Pharma and Bio Sciences, 5 (2): 835-840.

9. Krishnaswami, S., M. N. Narasimhanna, S. K. Suryanarayan & S. Kumararaj, 1973. “Manual of sericulture. Vol. 2. silkworm rearing, 68-71”. In Proceedings FAO Conference (24-27 June 1973, Rome, Italy), 350 pp.
10. Kurahashi, H., P. Atiwetin, S. Nagaoka, S. Miyata, S. Kitajima & Y. Sugimura, 2005. Absorption of mulberry root urease to the hemolymph of the silkworm, Bombyx mori. Journal of Insect Physiology, 51 (9): 1055-1061.
11. Leonardi, M. G., P. Marciani, P. G. Montorfono, S. Cappellozza, B. Giordana & G. Monticalli, 2001. Effects of fenoxycarb on leucine uptake and lipid composition of midgut brush border membrane in the silkworm, Bombyx mori (Lepidoptera: Bombycidae). Pesticide Biochemistry and Physiology, 70 (1): 42-51.
12. Malik, M. A. & F. A. M. Malik, 2009. Ontogenic changes in haemolymph biochemical composition in the Silkworm, Bombyx mori L under thermal stress. Academic Journal of Entomology, 2 (1): 16-21.
13. Megias, C., I. Cortes-Giraldo, J. Giron-Calle, J. Vioque & M. Alaiz, 2015. Determination of l-canavanine and other free amino acids in Vicia disperma (Fabaceae) seeds by precolumn derivatization using diethyl ethoxymethylenemalonate and reversed-phase high-performance liquid chromatography. Talanta, 131 (1): 95-98.
14. Mobley, H. L. T., M. D. Island & R. P. Hausinger, 1995. Molecular biology of microbial ureases. Microbiological Reviews, 59 (3): 451-480.
15. Monconduit, H. & B. Mauchamp, 1998. Effects of ultra-low doses of fenoxycarb on juvenile hormone regulated physiological parameters in the silkworm, Bombyx mori. Archives of Insect Biochemistry Physiology, 37 (2): 178-189.
16. Sahan, U., 2011. İpekböcekçiliği, İpekböceği Yetiştirme ve Islahı, Koza Üretimi, Ham İpek, Yumurta Üretimi, Hastalıklar ve Dut Yetiştirme. Dora Publisher, Bursa, Turkey, 150 pp.
17. Sahan, U., R. L. Buyukuysal & A. Erdemir, 2019. “A survey possible reasons of non-spinning syndrome in sericulture industry and its incidence in Turkey, 142-143”. 9th Bacsa International Conference “Sericulture preservation and revival - problems and prospects (7-12 April, Batumi, Georgia), 171 pp.
18. Savithri, G., P. Sujathamma & K. V. Asha, 2013. Silkworm Bombyx mori an economic insect. International Journal of Scientific Research, 2 (7): 535-537.
19. Sheliakina, M., V. Arkhypova, O. Soldatkin, O. Saiapina, B. Akatad & S. Dzyadevych, 2014. Urease-based ISFET biosensor for arginine determination. Talanta, 121 (4): 18-23.
20. Shen, W., W. Han, Y. Li, Z. Meng, L. Cai & L. Li, 2016. Development of chemical isotope labeling liquid chromatography mass spectrometry for silkworm hemolymph metabolomics. Analytica Chimica Acta, 942: 1-11.
21. Sirko, A. & R. Brodzik, 2000. Plant ureases: Roles and regulation. Acta Biochimica Polonica, 47 (4): 1189-95.
22. Sugimura, M., C. Hirayama & M. Nakamura, 2001. Selective transport of the mulberry leaf urease from the midgut into the larval hemolymph of the silkworm, Bombyx mori. Journal of Insect Physiology, 47 (10): 1133-1138.
23. Sumida, M., K. Haga, Y. Tanaka, J. Shimabukuro, M. Ichida & F. Matsubara, 1993. Developmental changes in urea in the hemolymph (determined by a urease-indophenol method) in hybrid strains of the silkworm, Bomhyx mori and the effect of starvation in the fifth instar larvae, fed an artificial diet on urea level in subsequent development. Comparative Biochemistry and Physiology, 105 (3): 563-570.
24. Sumida, M., Y. Yamada, Y. Tanaka, I. Shimabukuro, M. Ohnishi, H. Mori & F. Matsubara, 1990. Changes in urea in the haemolymph of the silkworm, silkworm, Bombyx mori in the fourth and the fifth larval instars and effects of starvation in the fifth instar on the level of urea in the pharate adults. Comparative Biochemistry and Physiology, 97 (3): 373-379.
25. Sumida, M., H. Yoshio, Y. Tanaka & F. Matsubara, 1995. Developmental changes in urea concentrations in the haemolymph of Daizo (T), an original strain of the silkworm, Bombyx mori reared on an artificial diet and on fresh mulberry leaves. Comparative Biochemistry and Physiology, 110 (1): 33-38.
26. Sun, X., H. Van-Der-Valk, H. Jiang, X. Wang, S. Yuan, Y. Zhang, I. Roessink & X. Gao, 2012. Development of a standard acute dietary toxicity test for the silkworm (Bombyx mori L.). Crop Protection, 42: 260-267.
27. Yamada, M., K. Nakamura & T. Inokuchi, 1983. Effects of diets on urea contents in the haemolymph and silk glandof the last instar larvae of the silkworm, Bombyx mori (Lepidoptera: Bombycidae). Applied Entomology and Zoology, 27 (2): 92-98.
28. Zhou, L., H. Li, F. Hao, N. Li, X. Liu, G. Wang, Y. Wang & H. Tang, 2015. Developmental changes for the hemolymph metabolome of silkworm (Bombyx mori L.). Journal of Proteome Research, 14 (5): 2331-2347.