Tropical Animal Health and Production

, Volume 50, Issue 1, pp 97–104 | Cite as

Evaluation of immunostimulatory and immunotherapeutic effects of tropical mushroom (Lentinus edodes) against eimeriasis in chicken

  • Muhammad Irfan UllahEmail author
  • Masood Akhtar
  • Mian Muhammad Awais
  • Muhammad Irfan Anwar
  • Kashfa Khaliq
Regular Articles


Mushrooms (Lentinus edodes) were processed for hot water (HWE), methanolic (ME), and polysaccharide (PSE) extracts. Polysaccharides were isolated through ion exchange (DEAE cellulose) and size exclusion (Sephadex G-100) chromatography. Monosaccharides including maltose (0.282%), glucose (0.113%), and mannose (0.451%) were identified, qualitatively and quantitatively, from the isolated polysaccharides through high-performance liquid chromatography. The whole study was divided into two experiments. Experiment 1 was meant for the evaluation of HWE and ME; whereas, experiment 2 was meant for the evaluation of PSE for immunostimulatory and immunotherapeutic activities. The cellular and humoral immune responses were demonstrated through lymphoproliferative response to phytohemagglutinin-P (PHA-P) and anti-body response to sheep red blood cells (SRBCs), respectively. The immunotherapeutic effects of these extracts were demonstrated against eimeriasis in terms of lesion scoring, oocysts per gram of droppings, and percent protection. Cell-mediated immune responses observed at 24, 48, and 72 h post-PHA-P injection were significantly higher (P < 0.05) in chickens administered with any of the three extracts (PSE, ME, and HWE), when compared with the controls. Humoral immune response in terms of anti-SRBCs anti-body titers was also observed higher in chickens administered with mushroom extracts. In the challenge experiment, significantly higher (P < 0.05) OPG and lesion scores were observed in controls as compared to the groups administered with mushroom extracts (HWE, ME, and PSE). Significantly higher (P < 0.05) percent protection against eimeriasis was observed in all groups administered with different extracts of L. edodes as compared to controls. In conclusion, L. edodes extracts showed immunostimulatory potential which persisted against eimeriasis in chicken.


Mushroom Lentinus edodes Eimeriasis Chicken Immunostimulation Immunotherapeutic 



Funds for this project were sponsored by Higher Education Commission (HEC), Islamabad, Pakistan, under the scheme of PhD indigenous scholarship Batch V already enrolled PhDs Pin No. 085-12327-Av5-22.

Compliance with ethical standards

All procedures in animal experimentation were conducted in accordance with the guidelines of institutional animal care and use committee after approval from advance studies research board University of Agriculture, Faisalabad, Pakistan.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Ahmad, I., 2008. Spatio-temporal variation in ecophysiological attributes of some medicinal plants indigenous to Soon Valley. (University of Agriculture, Faisalabad)Google Scholar
  2. Awais, M. M., Akhtar, M., Muhammad, F., Haq, A. U., Anwar, M. I., 2011. Immunotherapeutic effects of some sugar cane (Saccharum officinarum L.) extracts against coccidiosis in industrial broiler chickens. Experimental Parasitology, 128, 104–110.CrossRefGoogle Scholar
  3. Barros, L., Baptista, P., Correia, D. M., Casal, S., Oliveira, B. and Ferreira, I. C. F. R., 2007. Fatty acid and sugar compositions, and nutritional value of five wild edible mushrooms from Northeast Portugal. Food Chemistry, 105, 140–145.CrossRefGoogle Scholar
  4. Bohn, J. A. and BeMiller, J. N., 1995. (1-3)-β-D-Glucans as biological response modifiers: a review of structure – funtional activity relationships. Carbohydrate Polymer, 28, 3–14.CrossRefGoogle Scholar
  5. Burgh, M. A., 1978. Simple method for recording and analyzing serological data. Avian Diseases, 362–365.Google Scholar
  6. Chen, H. L., Li, D. F., Chang, B. Y., Gong, L. M., Piao, X. S., Yi, G. F. and Zhang, J. X., 2003. Effects of Lentinas on broiler splenocyte proliferation, interleukin-2 production, and signal transduction. Poultry Science, 760–766.Google Scholar
  7. Corrier, D. E., 1990. Comparison of phytohemagglutinin-induced cutaneous hypersensitivity reaction in the interdigital skin of broiler and layer chicks. Avian Diseases, 369–373.Google Scholar
  8. Cui, H. L., Chen, Y., Wang, S. S., Kai, G. Q. and Fang, Y. M., 2011. Isolation, partial characterization and immunomodulatory activities of polysaccharide from Morchella esculenta. Journal of the Science of Food and Agricultrue, 91, 2180–2185.Google Scholar
  9. Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A. and Smith, F., 1956. Colorimetric methods for determination of sugars and related substances. Analytical Chemistry, 28, 350–356.CrossRefGoogle Scholar
  10. Guillamon, E., Gracia-Lafuente, A., Lozano, M., Arrigo, M. D., Rostagno, M. A., Villares, A. and Alfredo, J., 2010. Edible mushrooms: Role in the prevention of cardiovascular diseases. Fitoterapia, 81, 715–723.CrossRefGoogle Scholar
  11. Guo, F. C., Kwakkel, R. P., Williams, B. A., Suo, X., Li, W. K. and Verstegen, M. W. A., 2005. Coccidiosis immunization: Effects of mushroom and herb polysaccharides on immune responses of chicken infected with Eimeria tenella. Avian Diseases, 49, 70–73.CrossRefGoogle Scholar
  12. Guo, F. C., Savelkoul, H. F. J., Kwakkel, R. P., Williams, B. A. and Verstegen, M. W. A., 2003. Immunoactive, medicinal properties of mushroom and herb polysaccharides and their potential use in chicken diets. World’s Poultry Science Journal, 59, 427–440.CrossRefGoogle Scholar
  13. Guo, F. C., Williams, B. A., Kwakkel, R. P., Li, H. S. Li, X. P., Luo, J. Y., Li, W. K. and Verstegen, M. W. A., 2004. Effects of mushroom and herb polydaccharides, as alternatives for antibiotic on the cecal microbial ecosystem in broiler chickens. Poultry Science, 83, 175–182.CrossRefGoogle Scholar
  14. Han, S. R., Cho, C., Lee, Y. and Yoo, H., 2009. Antimetastatic and immunomodulatory effect of water extracts from various mushrooms. Journal of Acupuncture and Meridian Studies, 2, 218–227.CrossRefGoogle Scholar
  15. Jackson, J., Hurley, S. L. and Anike, F., 2012. Effect of different mushrooms fed to eimeria-challenged broilers on rearing performance. International Journal of Poultry Science, 11, 433–437.CrossRefGoogle Scholar
  16. Johnson, J. and Reid, W. M., 1970. Anticoccidial drugs: Lesion scoring techniques in battery and floor pen experiments with chickens. Experimental Parasitology, 28, 30–36.CrossRefGoogle Scholar
  17. Kaneno, R., Fontanari, L. M., Santos, S. A., Di Stasi, L. C., Rodrigues-Filho, E. and Eira, A. F., 2004. Effects of extracts from Brazilian sun-mushroom (Agaricus blazei) on the NK activity and lymphoproliferative responsiveness of Ehrlich tumor-bearing mice. Food and Chemical Toxicology, 42, 909–916.CrossRefGoogle Scholar
  18. Levine, N. D. and others, 1961. Protozoan parasites of domestic animals and of man (Minneapolis, Minn: Burgess Publishing Company).Google Scholar
  19. Lindequest, U., Niedermeye, T. H. J. and Ju, W., 2005. The pharmacological potential of mushrooms. Evidence-Based Complementary and Alternative Medicine, 2, 285–299.CrossRefGoogle Scholar
  20. Maji, P. K., Sen, I. K., Devi K. S. P., Maiti T. K., Sikdar S. R. and Islam, S. S., 2013. Structural elucidation of a biologically active heteroglycan isolated from a hybrid mushroom of Pleurotus florida and Lentinula edodes. Carbohydrate Research, 368, 22–28.CrossRefGoogle Scholar
  21. Martin, A. G., Danforth, H. D., Brta, J. R. and Fernando, M. A., 1997. Analysis of immunological cross-protection and sensitivities to anticoccidial drugs among five geographical and temporal strains of Eimeria maxima. International Journal for Parasitology, 27, 527–533.CrossRefGoogle Scholar
  22. Moro, C., Palacios, I., Lozano, M., Arrigo, M. D., Guillamon, E. Villares, A., Martinez, J. A. and Garcia-lafuente, A., 2012. Anti-inflammatory activity of methanolic extracts from edible mushrooms in LPS activated RAW 264.7 macrophages. Food Chemistry, 130, 350–355.CrossRefGoogle Scholar
  23. Osborn, H. M. I., Lochey, F., Mosley, L., and Read, D., 1999. Analysis of polysaccharides and monosaccharides in the root mucilage of maize (Zea mays L.) by gas chromatography. Journal of Chromatography A 831(2):267–276.Google Scholar
  24. Ouzouni, P. K., Petridis, D., Koller, W. D. and Riganakos, K. A., 2009. Nutritional value and metal content of wild mushrooms collected from West Macedonia and Epirus Greece. Food Chemistry, 115, 1575–1580.CrossRefGoogle Scholar
  25. Qureshi, M. A. and Havenstein, G. B., 1994. A comparison of the immune performance of a 1991 commercial broiler with a 1957 randombred strain when fed “typical” 1957 and 1991 broiler diets Poultry Science, 73, 1805–1812.CrossRefGoogle Scholar
  26. Rowan, N. J., Smith, J. E. and Sullivan, R., 2003. Immunomodulatory activities of mushroom glucans and polysaccharide-protein complexes in animals and humans (a review). International Journal of Medicinal Mushrooms, 5.Google Scholar
  27. Sarangi, I., Ghosh, D., Bhutia, S. K., Mallick, S. K. and Maiti, T. K., 2006. Anti-tumor and immunomodulating effects of Pleurotus ostreatus mycelia-derived proteoglycans. International Immunopharmacology, 6, 1287–1297.CrossRefGoogle Scholar
  28. Shamtsyan, M., Konusova, V., Maksimova, Y., Goloshchev, A., Panchenko, A., Simbirtsev, A., Petrishchev, N. and Denisova, N., 2004. Immunomodulating and anti-tumor action of extracts of several mushrooms. Journal of Biotechnology, 113, 77–83.CrossRefGoogle Scholar
  29. Smiderle, F. R., Olsen, L. M., Carbonero, E. R., Baggio, C. H., Feritas, C. S., Marcon, R., Santos, A. R. S., Gorin, P. A. J. and Iacomini, M., 2008. Anti-inflammatory and analgesic properties in a rodent model of a (1-3) (1-6)-linked β-glucan isolated from Pleurotus pulmonaris. European Journal of Pharmacology, 597, 86–91.CrossRefGoogle Scholar
  30. Ullah, M. I., Akhtar, M., Iqbal, Z., Shahid, M. and Awais, M. M., 2015. Immunomodulating and antiprotozoal effects of different extracts of oyster culinary-medicinal mushroom Pleurotus ostreatus (Higher Basidiomycetes) against coccidiosis in broiler. International Journal of Medicinal Mushrooms, 17, 309–317.CrossRefGoogle Scholar
  31. Xu, X., Yan, H., Tang, J., Chen, J. and Zhang, X., 2014. Polysaccharides in Lentinus edodes: isolation, structure, immunomodulating activity and future prospective. Critical Review in Food Science and Nutrition, 54, 474–487.CrossRefGoogle Scholar
  32. Xue, M. and Meng, X. S., 1996. Review on research progress and prosperous of immune activities of bio-active polysaccharides. Journal of Traditional Veterinary Medicine, 3, 15–18Google Scholar
  33. Yang, J. H., Lin, H. C. and Mau, J. L., 2002. Antioxidant properties of several commercial mushrooms. Food Chemistry, 77, 229–235.CrossRefGoogle Scholar
  34. Yang, Q., Wang, S., Xie, Y., Sun, J. and Wang, J., 2010. HPLC analysis of Ganoderma lucidum polysaccharides and its effect on antioxidant enzymes activity and Bax, BCL-2 expression. International Journal of Biological Macromolecules, 46, 167–172.CrossRefGoogle Scholar
  35. Zhao, L., Dong, Y., Chen, G. and Hu, Q., 2010. Extraction, purification, characterization and antitumor activity of polysaccharides from Ganoderma lucidum. Carbohydrate Polymer, 80, 783–789.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  • Muhammad Irfan Ullah
    • 1
    • 2
    Email author
  • Masood Akhtar
    • 1
    • 2
  • Mian Muhammad Awais
    • 1
  • Muhammad Irfan Anwar
    • 1
  • Kashfa Khaliq
    • 3
  1. 1.Department of Pathobiology, Faculty of Veterinary SciencesBahauddin Zakariya UniversityMultanPakistan
  2. 2.Immunoparasitology Laboratory, Department of Parasitology, Faculty of Veterinary SciencesUniversity of AgricultureFaisalabadPakistan
  3. 3.Department of MicrobiologyGovernment College UniversityFaisalabadPakistan

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