Immunostimulant Effects of Geophyte Plant Extract on Non-specific Defence Mechanisms of Rainbow Trout (Oncorhynchus mykiss)

Abstract

The aim of this study was to determine the immunostimulant effects of geophyte plant extract on non-specific defence mechanisms of rainbow trout (Oncorhynchus mykiss). For this purpose, Muscari comosum was collected in Muğla region and extracted in ethanol. Then, The plant extract applied into fish by intraperitonal injection in two different concentrations (0.5mg/ fish and 2.0 mg/fish). The average fish weight was 140 g. Following the injection on the 1st, 7th, 14th, 21st, 28th days the blood and serum samples were collected from fish in each group and examined for various parameters including percentage of hematocrit, the counts of nitroblue tetrazolium (NBT) positive neutrophils, total leukocyte counts, percentage of white blood cells and serum lysozyme activity. The results indicated that the counts of NBT (+)  neutrophils, percentage of monocyte and neutophil and total leukocyte counts increased in the group with injected 0.5mg plant extract /fish compared to control group (P<0.05).  It has been revealed that this dose of M. comosum can be suggested to use to enhance non-specific immune system for rainbow trout aquaculture. However, immunostimulant effects of oral administration of this plant extract in rainbow trout are needed to be determine in future studies.

Keywords

• Fish,immune system,Muscari comosum,Tassel hyacinth,blood

Gökkuşağı Alabalığı (Oncorhynchus mykiss)’nın Spesifik Olmayan Savunma Mekanizması Üzerine Geofit Bitki Ekstraktının İmmunostimulant Etkisi

Abstract

Bu çalışmada amacımız, gökkuşağı alabalığının spesifik olmayan savunma mekanizması üzerine geofit bitki ektraktının immunostimulant etkisinin belirlenmesidir. Bu amaçla geofit bir bitki olan M. comosum Muğla bölgesinden toplandı ve etanolde ekstrakte edildi. Daha sonra farklı dozlarında (0,5 mg/balık ve 2,0 mg/balık) balıklara intraperitonal olarak uygulandı. Balıklar ortalama 140 g ağırlığa sahipti. Deneme süresince uygulamayı takiben 1, 7, 14, 21, 28. günlerde herbir gruptaki balıktan kan ve serum örnekleri alınarak farklı parametreler (hematokrit yüzdesi, nitroblue tetrazolium (NBT) pozitif nötrofil sayısı, toplam lökosit sayısı, beyaz kan hücrelerinin yüzdesi ve serum lizozim aktivitesi) açısından incelendi. Bu çalışmanın sonucunda NBT pozitif nötrofillerin sayısı, monosit ve nötrofil yüzdeleri ve toplam lökosit sayısı 0,5 mg/balık dozunda injeksiyon yapılan grupta kontrol grubuna kıyasla artış gösterdiği (P<0,05) belirlendi.  Bu nedenle gökkuşağı alabalığı yetiştiriciliğinde spesifik olmayan immun sistemi uyarmak için bu dozdaki M. comosum’un kullanılması önerilebilir. Ancak, bitki ekstraktının oral yolla yeme ilave edilerek balıklara uygulanması ile immunostimulant etkisinin belirlenmesi için gelecekte yeni çalışmalara ihtiyaç duyulmaktadır.

Keywords

• Balık,immun sistem,Muscari comosum,arap sümbülü,kan

References

1. Aly SM, Mohamed MF. 2010. Echinacea purpurea and Allium sativum as immunostimulants in fish culture using Nile tilapia (Oreochromis niloticus). J Anim Physiol Anim Nutr (Berl). 94 (5), e31-e39 doi: 10.1111/j.1439-0396.2009.00971.x
2. Anderson DP, Moritomo T, Grooth RD. 1992. Neutrophile, glass-adherent, nitroblue tetrazolium assay gives early indication of immunization effectiveness in rainbow trout. Vet. Immunol.Immunopathol.30(4):419-429. doi:10.1016/0165-2427(92)90110-C
3. Awad E, Awaad A. 2017. Role of medicinal plants on growth performance and immune status in fish. Fish Shellfish Immunol. 67(1):40-54. doi: 10.1016/j.fsi.2017.05.034
4. Awad E, Austin D, Lyndon AR. 2013. Effect of black cumin seed oil (Nigella sativa) and nettle extract (Quercetin) on enhancement of immunity in rainbow trout, Oncorhynchus mykiss (Walbaum). Aquaculture 388-391(1):193–197 doi: 10.1016/j.aquaculture.2013.01.008
5. Baba E, Acar Ü, Öntaş C, Kesbiç OS, Yilmaz S. 2016. The use of Avena sativa extract against Aeromonas hydrophila and its effect on growth performance, hematological and immunological parameters in common carp (Cyprinus carpio). Ital. J. Anim. Sci. 15 (2): 325-333. doi: 10.1080/1828051X.2016.1185977
6. Baba E, Uluköy G, Mammadov R. 2014. Effects of Muscari comosum Extract on Nonspecific Immune Parameters in Gilthead Seabream, Sparus aurata (L. 1758). J World Aquacult. Soc. 45 (2): 173-182 doi: 10.1111/jwas.12100
7. Christybapita D, Divyagnaneswari M, Michael RD. 2007. Oral administration of Eclipta alba leaf aqueous extract enhances the non-specific immune responses and disease resistance of Oreochromis mossambicus. Fish Shellfish Immunol. 23(4):840-852 doi: 10.1016/j.fsi.2007.03.010
8. Deivasigamani B, Subramanian V. 2016. Applications of Immunostimulants in Aquaculture: A Review. Int. J. Curr. Microbiol. App. Sci. 5(9): 447-453. doi: 10.20546/ijcmas.2016.509.048

9. Dügenci SK, Arda N, Candan A. 2003. Some medicinal plants as immunostimulant for fish. J Ethnopharmacol.88 (1), 99-106 doi: 10.1016/S0378-8741(03)00182-X
10. Engstad RE, Robertsen B, Frivold E. 1992. Yeast glucan induces increase in lysozyme and complement-mediated haemolytic activity in Atlantic salmon blood. Fish Shellfish Immunol. 2 (4): 287-297 doi: 10.1016/S1050-4648(06)80033-1
11. Immanue G, Uma RP, Iyapparaj P, Citarasu T, Punitha Peter SM, Michael Babu M, Palavesam A. 2009. Dietary medicinal plant extracts improve growth, immune activity and survival of tilapia Oreochromis mossambicus. J Fish Biol 74 (7): 1462-1475 doi: 10.1111/j.1095-8649.2009.02212.x
12. Lee TH, Qiu F, Waller GR, Chou CH. 2000. Three new flavonol galloylglycosides from leaves of Acacia confusa. J. Nat. Prod. 63(5): 710-712 doi: 10.1021/np990482w
13. Logambal SM, Venkatalakshmi S, Michael RD. 2000. Immunostimulatory effect of leaf extract of Ocimum sanctum Linn. in Oreochromis mossambicus (Peters). Hydrobiologia, 430 (1-3):113-120. doi: 10.1023/A:1004029332114
14. Loizzo MR, Tundis R, Menichini F. 2010. Chelating, antioxidant and hypoglycaemic potential of Muscari comosum (L.) Mill. bulb extracts. Int J Food Sci Nutr. 61(8):780-791. doi: 10.3109/09637486.2010.482521
15. Magnadottir B. 2006. Innate immunity of fish (Overview). Fish Shellfish Immunol. 20 (2): 137-151. doi: 10.1016/j.fsi.2004.09.006
16. Mehana EE, Rahmani AH, Aly SM. 2015. Immunostimulants and Fish Culture: An Overview. Annu Res Rev Biol. 5 (6):477-489 doi: 10.9734/ARRB/2015/9558
17. Nasrabadi, M, Halimi M, Nadaf M. 2013. Phytochemical screening and chemical composition of extract of Muscari neglectum. Middle-East J Sci Res 14(4):566–569. doi: 10.5829/idosi.mejsr.2013.14.4.71191
18. Pieroni, A, Janiak V, Durr CM, Ludeke S, Trachsel E, Heinrich M. 2002. In vitro antioxidant activity of non-cultivated vegetables of ethnic Albanians in southern Italy. Phytother. Res. 16 (5):467–473. doi: 10.1002/ptr.1243
19. Plumb, J. A., and Hanson, LA. 2011. Health Maintenance and Principal Microbial Diseases of Cultured Fishes. Hoboken, NJ: Wiley-Blackwell. doi: 10.1002/9780470958353
20. Pratheepa V, Ramesh S, Sukumaran N. 2010. Immunomodulatory effect of Aegle marmelos leaf extract on freshwater fish Cyprinus carpio infected by bacterial pathogen Aeromonas hydrophila. Pharm. Biol.48 (11): 1224-1239 doi: 10.3109/13880201003713598
21. Schaperclaus W, Kulow H, Schreckenbach K. (1991) Hematological and serological technique. In: Fish Disease, Vol. 1, 2nd edn, ed. by V.S.Kothekar), pp. 71–108. Gulab primlani, Oxonian Press, New Delhi, India.
22. Steinhagen D, Kruse P, Körting W. 1990. Some heamatological observations on carp, Cyprinus carpio L. experimentally infected with Trypanoplasma borreli Laveron. J Fish Dis. 13(2) :157–162 doi: 10.1111/j.1365-2761.1990.tb00768.x
23. Sudhakaran DS, Srirekha P, Devasree LD, Premsingh S, Michael RD. 2006. Immunostimulatory effect of Tinospora cordifolia Miers leaf extract in Oreochromis mossambicus. Indian J Exp Biol. 44 (9):726-732 URL: https://www.ncbi.nlm.nih.gov/pubmed/16999027
24. Sharma A, Deo AD, Riteshkumar ST, Chanu TI, Das A. 2010. Effect of Withania somnifera (L. Dunal) root as a feed additive on immunological parameters and disease resistance to Aeromonas hydrophila in Labeo rohita (Hamilton) fingerlings. Fish Shellfish Immunol. 29 (3):508-512 doi: 10.1016/j.fsi.2010.05.005
25. Tanker M, Tanker N. 1991. Farmakognozi Cilt 1. Ankara Universitesi, Eczacılık Fakültesi yayını, No:66, Ankara. (In Turkish) URL: http://kitaplar.ankara.edu.tr/dosyalar/pdf/313.pdf
26. Villa F, Borgonovo G, Cappitelli F, Giussani B, Bassoli A. 2012. Sub-lethal concentrations of Muscari comosum bulb extract suppress adhesion and induce detachment of sessile yeast cells. Biofoling 28(10):1107–1117. doi: 10.1080/08927014.2012.734811