Cell Stress and Chaperones

, Volume 23, Issue 5, pp 1069–1078 | Cite as

Protective effects of zymosan on heat stress-induced immunosuppression and apoptosis in dairy cows and peripheral blood mononuclear cells

  • Yuhang Sun
  • Jin Liu
  • Gengping Ye
  • Fang Gan
  • Mohammed Hamid
  • Shengfa Liao
  • Kehe Huang
Original Paper


Dairy cows exposed to heat stress (HS) show decreased performance and immunity, but increased heat shock protein expressions and apoptosis. Zymosan, an extract from yeast cell walls, has been shown to modulate immune responses and defense against oxidative stress. However, few literatures are available about the effects of zymosan on immune responses and other parameters of the dairy cows under HS. Here, both primary peripheral blood mononuclear cell (PBMC) and dairy cow models were established to assess the effects of zymosan on performance, immunity, heat shock protein, and apoptosis-related gene expressions of dairy cows under HS. In vitro study showed that proliferation, IL-2 production, and Bcl-2/Bax-α ratio of cow primary PBMC were reduced, whereas hsp70 mRNA and protein expressions, as well as Annexin V-bing, were increased when PBMCs were exposed to heat. In contrast, zymosan significantly reversed these above changes induced by the HS. In the in vivo study, 40 Holstein dairy cows were randomly selected and assigned into zymosan group (supplemental zymosan; n = 20) and control group (no supplemental zymosan; n = 20). The results showed that zymosan improved significantly the dry matter intake and milk yield, increased IgA, IL-2, and tumor necrosis factor-α (TNF-α) contents in sera, as well as hepatic Bcl-2/Bax-α ratio, but decreased respiration rate and hepatic hsp70 expressions in the dairy cows under HS. Taken together, zymosan could alleviate HS-induced immunosuppression and apoptosis and improve significantly the productive performance and immunity of dairy cows under HS.


Zymosan Heat stress Dairy cows Peripheral blood mononuclear cells Immunity Apoptosis 


Funding information

This work was supported by the National Key R & D Program (2016YFD0501203), the National Natural Science Foundation of China (No. 31472253 and 31772811), and the Priority Academic Program Development of Jiangsu Higher Education Institutions (Jiangsu, China).

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interests.

Supplementary material

12192_2018_916_Fig6_ESM.png (960 kb)
Fig. S1

Daily air temperature in the barn during this study. This study was performed for 28 days, during which time the air temperature was recorded 3 times daily at 0700, 1400 and 2100 h, respectively. (PNG 31 kb)

12192_2018_916_MOESM1_ESM.tiff (162 kb)
High Resolution (TIFF 161 kb)


  1. Bhanuprakash V, Singh U, Sengar G, Sajjanar B, Bhusan B, Raja TV, Alex R, Kumar S, Singh R, Ashish Kumar, Alyethodi RR, Kumar S, Deb R (2016) Differential effect of thermal stress on HSP70 expression, nitric oxide production and cell proliferation among native and crossbred dairy cattle. J Therm Biol 59:18–25. CrossRefPubMedGoogle Scholar
  2. Brown BM, Stallings JW, Clay JS, Rhoads ML (2016) Correction: periconceptional heat stress of Holstein dams is associated with differences in daughter milk production and composition during multiple lactations. PLoS One 11:e0150049. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Carroll JA, Burdick NC, Chase CC Jr, Coleman SW, Spiers DE (2012) Influence of environmental temperature on the physiological, endocrine, and immune responses in livestock exposed to a provocative immune challenge. Domest Anim Endocrinol 43:146–153. CrossRefPubMedGoogle Scholar
  4. Carter LL, Zhang X, Dubey C, Rogers P, Tsui L, Swain SL (1998) Regulation of T cell subsets from naive to memory. J Immunother 21:181–187CrossRefPubMedGoogle Scholar
  5. Dahl GE, Tao S, Monteiro APA (2016) Effects of late-gestation heat stress on immunity and performance of calves. J Dairy Sci 99:3193–3198. CrossRefPubMedGoogle Scholar
  6. De Rensis F, Garcia-Ispierto I, Lopez-Gatius F (2015) Seasonal heat stress: clinical implications and hormone treatments for the fertility of dairy cows. Theriogenology 84:659–666. CrossRefPubMedGoogle Scholar
  7. Deja M, Ahlers O, Macguill M, Wust P, Hildebrandt B, Riess H, Kerner T (2010) Changes in hepatic blood flow during whole body hyperthermia. Int J Hyperth 26:95–100. CrossRefGoogle Scholar
  8. Do Amaral B, Connor E, Tao S, Hayen M, Bubolz J, Dahl G (2011) Heat stress abatement during the dry period influences metabolic gene expression and improves immune status in the transition period of dairy cows. J Dairy Sci 94:86–96CrossRefPubMedGoogle Scholar
  9. Du J et al (2017) Zymosan-a protects the hematopoietic system from radiation-induced damage by targeting TLR2 signaling pathway. Cell Physiol Biochem 43:457–464. CrossRefPubMedGoogle Scholar
  10. Elvinger F, Hansen PJ, Natzke RP (1991) Modulation of function of bovine polymorphonuclear leukocytes and lymphocytes by high temperature in vitro and in vivo. Am J Vet Res 52:1692–1698PubMedGoogle Scholar
  11. Fabris TF, Laporta J, Corra FN, Torres YM, Kirk DJ, McLean DJ, Chapman JD, Dahl GE (2017) Effect of nutritional immunomodulation and heat stress during the dry period on subsequent performance of cows. J Dairy Sci 100:6733–6742. CrossRefPubMedGoogle Scholar
  12. Frasnelli ME, Tarussio D, Chobaz-Peclat V, Busso N, So A (2005) TLR2 modulates inflammation in zymosan-induced arthritis in mice. Arthritis Res Ther 7:R370–R379. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Gan F, Zhang Z, Hu Z, Hesketh J, Xue H, Chen X, Hao S, Huang Y, Cole Ezea P, Parveen F, Huang K (2015) Ochratoxin A promotes porcine circovirus type 2 replication in vitro and in vivo. Free Radic Biol Med 80:33–47. CrossRefPubMedGoogle Scholar
  14. Jiang LI, Sternweis PC, Wang JE (2013) Zymosan activates protein kinase A via adenylyl cyclase VII to modulate innate immune responses during inflammation. Mol Immunol 54:14–22. CrossRefPubMedGoogle Scholar
  15. Ju XH, Xu HJ, Yong YH, An LL, Jiao PR, Liao M (2014) Heat stress upregulation of toll-like receptors 2/4 and acute inflammatory cytokines in peripheral blood mononuclear cell (PBMC) of Bama miniature pigs: an in vivo and in vitro study. Animal 8:1462–1468. CrossRefPubMedGoogle Scholar
  16. Kadokawa H, Sakatani M, Hansen PJ (2012) Perspectives on improvement of reproduction in cattle during heat stress in a future Japan. Anim Sci J 83:439–445. CrossRefPubMedGoogle Scholar
  17. Karimi MT, Ghorbani GR, Kargar S, Drackley JK (2015) Late-gestation heat stress abatement on performance and behavior of Holstein dairy cows. J Dairy Sci 98:6865–6875. CrossRefPubMedGoogle Scholar
  18. Kelley DW, Osborne CA, Evermann JF, Parish SM, Gaskins CT (1982) Effects of chronic heat and cold stressors on plasma immunoglobulin and mitogen-induced blastogenesis in calves. J Dairy Sci 65:1514–1528CrossRefPubMedGoogle Scholar
  19. Kishore A, Sodhi M, Kumari P, Mohanty AK, Sadana DK, Kapila N, Khate K, Shandilya U, Kataria RS, Mukesh M (2014) Peripheral blood mononuclear cells: a potential cellular system to understand differential heat shock response across native cattle (Bos indicus), exotic cattle (Bos taurus), and riverine buffaloes (Bubalus bubalis) of India. Cell Stress Chaperones 19:613–621. CrossRefPubMedGoogle Scholar
  20. Kogan G et al (2005) Antioxidant properties of yeast (1→3)-β-d-glucan studied by electron paramagnetic resonance spectroscopy and its activity in the adjuvant arthritis. Carbohydr Polym 61:18–28. CrossRefGoogle Scholar
  21. Lacetera N, Bernabucci U, Scalia D, Ronchi B, Kuzminsky G, Nardone A (2005) Lymphocyte functions in dairy cows in hot environment. Int J Biometeorol 50:105–110. CrossRefPubMedGoogle Scholar
  22. Li H, Gonnella P, Safavi F, Vessal G, Nourbakhsh B, Zhou F, Zhang GX, Rostami A (2013) Low dose zymosan ameliorates both chronic and relapsing experimental autoimmune encephalomyelitis. J Neuroimmunol 254:28–38. CrossRefPubMedGoogle Scholar
  23. Li L, Wu J, Luo M, Sun Y, Wang G (2016) The effect of heat stress on gene expression, synthesis of steroids, and apoptosis in bovine granulosa cells. Cell Stress Chaperones 21:467–475. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Liu J, Ye G, Zhou Y, Liu Y, Zhao L, Liu Y, Chen X, Huang D, Liao SF, Huang K (2014) Feeding glycerol-enriched yeast culture improves performance, energy status, and heat shock protein gene expression of lactating Holstein cows under heat stress. J Anim Sci 92:2494–2502. CrossRefPubMedGoogle Scholar
  25. Liu X, Li H, Lu A, Zhong Y, Hou X, Wang N, Jia D, Zan J, Zhao H, Xu J, Liu F (2012) Reduction of intestinal mucosal immune function in heat-stressed rats and bacterial translocation. Int J Hyperth 28:756–765. CrossRefGoogle Scholar
  26. Mishra A, Hooda OK, Singh G, Meur SK (2011) Influence of induced heat stress on HSP70 in buffalo lymphocytes. J Anim Physiol Anim Nutr 95:540–544. CrossRefGoogle Scholar
  27. Modrow JH, Preusse-Prange A, Meyer P, Harder M, Schwark T, von Wurmb-Schwark N (2012) Highly reliable quantification of proteins such as members of the HSP70 superfamily based on the grey scale index via immune detection stained bands on a western blot. Forensic Sci Int 222:256–258. CrossRefPubMedGoogle Scholar
  28. Monteiro AP, Tao S, Thompson IM, Dahl GE (2014) Effect of heat stress during late gestation on immune function and growth performance of calves: isolation of altered colostral and calf factors. J Dairy Sci 97:6426–6439. CrossRefPubMedGoogle Scholar
  29. Mosser DD, Caron AW, Bourget L, Denis-Larose C, Massie B (1997) Role of the human heat shock protein hsp70 in protection against stress-induced apoptosis. Mol Cell Biol 17:5317–5327CrossRefPubMedPubMedCentralGoogle Scholar
  30. Mymrikov EV, Daake M, Richter B, Haslbeck M, Buchner J (2017) The chaperone activity and substrate spectrum of human small heat shock proteins. J Biol Chem 292:672–684. CrossRefPubMedGoogle Scholar
  31. Nakamura H, Nagase H, Ogino K, Hatta K, Matsuzaki I (2001) Involvement of central, but not placental corticotropin releasing hormone (CRH) in heat stress induced immunosuppression during pregnancy. Brain Behav Immun 15:43–53. CrossRefPubMedGoogle Scholar
  32. Polsky L, von Keyserlingk MAG (2017) Invited review: effects of heat stress on dairy cattle welfare. J Dairy Sci 100:8645–8657. CrossRefPubMedGoogle Scholar
  33. Quinteiro-Filho W et al (2010) Heat stress impairs performance parameters, induces intestinal injury, and decreases macrophage activity in broiler chickens. Poult Sci 89:1905–1914CrossRefPubMedGoogle Scholar
  34. Radons J (2016) The human HSP70 family of chaperones: where do we stand? Cell Stress Chaperones 21:379–404. CrossRefPubMedPubMedCentralGoogle Scholar
  35. Ross GD, Cain JA, Lachmann PJ (1985) Membrane complement receptor type three (CR3) has lectin-like properties analogous to bovine conglutinin as functions as a receptor for zymosan and rabbit erythrocytes as well as a receptor for iC3b. J Immunol 134:3307–3315PubMedGoogle Scholar
  36. Rout PK, Kaushik R, Ramachandran N (2016) Differential expression pattern of heat shock protein 70 gene in tissues and heat stress phenotypes in goats during peak heat stress period. Cell Stress Chaperones 21:645–651. CrossRefPubMedPubMedCentralGoogle Scholar
  37. Sato M, Sano H, Iwaki D, Kudo K, Konishi M, Takahashi H, Takahashi T, Imaizumi H, Asai Y, Kuroki Y (2003) Direct binding of toll-like receptor 2 to zymosan, and zymosan-induced NF-B activation and TNF-secretion are down-regulated by lung collectin surfactant protein A. J Immunol 171:417–425. CrossRefPubMedGoogle Scholar
  38. Sellins KS, Cohen JJ (1991) Hyperthermia induces apoptosis in thymocytes. Radiat Res 126:88–95CrossRefPubMedGoogle Scholar
  39. Sherwood ER, Williams DL, McNamee RB, Jones EL, Browder IW, Di Luzio NR (1987) Enhancement of interleukin-1 and interleukin-2 production by soluble glucan. Int J Immunopharmacol 9:261–267CrossRefPubMedGoogle Scholar
  40. Shwartz G, Rhoads ML, VanBaale MJ, Rhoads RP, Baumgard LH (2009) Effects of a supplemental yeast culture on heat-stressed lactating Holstein cows1. J Dairy Sci 92:935–942. CrossRefPubMedGoogle Scholar
  41. Sodja C, Brown DL, Walker PR, Chaly N (1998) Splenic T lymphocytes die preferentially during heat-induced apoptosis: NuMA reorganization as a marker. J Cell Sci 111:2305–2313PubMedGoogle Scholar
  42. Song JS, Kim YJ, Han KU, Yoon BD, Kim JW (2015) Zymosan and PMA activate the immune responses of Mutz3-derived dendritic cells synergistically. Immunol Lett 167:41–46. CrossRefPubMedGoogle Scholar
  43. Tao S, Monteiro AP, Thompson IM, Hayen MJ, Dahl GE (2012) Effect of late-gestation maternal heat stress on growth and immune function of dairy calves. J Dairy Sci 95:7128–7136. CrossRefPubMedGoogle Scholar
  44. Taghavi M, Mortaz E, Khosravi A, Vahedi G, Folkerts G, Varahram M, Kazempour-Dizaji M, Garssen J, Adcock IM (2018) Zymosan attenuates melanoma growth progression, increases splenocyte proliferation and induces TLR-2/4 and TNF-alpha expression in mice. J Inflamm 15:5. CrossRefGoogle Scholar
  45. Umehara K, Hoshikawa M, Tochio N, Tate SI (2018) Substrate binding switches the conformation at the lynchpin site in the substrate-binding domain of human hsp70 to enable allosteric interdomain communication. Molecules 23.
  46. Wang L, Wang Z, Zou H, Peng Q (2016) Yeast culture and vitamin E supplementation alleviates heat stress in dairy goats. Asian Australas J Anim Sci 29:814–822. CrossRefPubMedGoogle Scholar
  47. Xu C, Sun Y, Yang W, Zhang H, Xia C (2016) Urine proteomics analysis of dairy cows with fatty liver. Pak Vet J 36:109–111Google Scholar
  48. Zhang P, Leu JI, Murphy ME, George DL, Marmorstein R (2014) Crystal structure of the stress-inducible human heat shock protein 70 substrate-binding domain in complex with peptide substrate. PLoS One 9:e103518. CrossRefPubMedPubMedCentralGoogle Scholar
  49. Zhuang T, Xu H, Hao S, Ren F, Chen X, Pan C, Huang K (2015) Effects of selenium on proliferation, interleukin-2 production and selenoprotein mRNA expression of normal and dexamethasone-treated porcine splenocytes. Res Vet Sci 98:59–65. CrossRefPubMedGoogle Scholar

Copyright information

© Cell Stress Society International 2018

Authors and Affiliations

  1. 1.College of Veterinary MedicineNanjing Agricultural UniversityNanjingChina
  2. 2.Shanghai Bright Holstein Co., Ltd.ShanghaiChina
  3. 3.Department of Animal and Dairy SciencesMississippi State UniversityStarkvilleUSA

Personalised recommendations