Aquaculture International

, Volume 27, Issue 1, pp 105–123 | Cite as

Environmental stress tolerance and immune response for the small abalone hybrids

  • Weiwei YouEmail author
  • Bo Wang
  • Xuan Luo
  • Caihuan Ke


Recently, mass mortality affected the cultured small abalone, Haliotis diversicolor diversicolor, which was the dominant cultured abalone species in southern China. Prior studies revealed that survivorship varied significantly between different stocks and crosses. However, the immunological basis for differences in susceptibility has not been well understood to date. Herein, low temperature, air exposure tolerance tests, and pull-off force measurement were assessed in the three groups (Japan, Taiwan, and their Hybrid stock). The results showed that the critical thermal minimum (CTMin) at 50% was 15.6 °C for the Taiwan stock, 12.1 °C for the Japan stock, and 13.2 °C for the Hybrid stock. Upon air exposure challenge, 100% abalones from the Taiwan group died after 8 h at 24 °C, while the survival rate in the Japan and Hybrid groups were 37.8% and 29.4%, respectively. The detachment stress for the Japan group was 42.3 kPa, which was 2.78-fold and 1.43-fold higher compared to the Taiwan and Hybrid groups, respectively. Variation in susceptibility to disease may be based on the effectiveness of the innate immune responses. Therefore, total hemocyte count, phagocytosis, respiratory burst, superoxide dismutase activity, acid phosphatase activity, alkaline phosphatase activity, and myeloperoxidase activity were determined for the healthy abalones in each group. Positive mid-parent heterosis on immunological parameters was consistent with the prior knowledge on the positive mid-parent heterosis for survival rate, which indicated the improvement on immune reaction and disease resistance through hybridization methods. The current study will be useful in efficient design of breeding programs for the development of sustainable abalone aquaculture.


Haliotis diversicolor diversicolor Heterosis Immune response Pull-off force Stress tolerance 



Critical thermal minimum


Reactive oxygen species


Acid phosphatase


Alkaline phosphatase


Superoxide dismutase


Weight loss rate


Total hemocyte count


First fluorescence


Phagocytic rate


2′,7′-Dichlorfluorescein diacetate








Sodium-dodecyl sulfate


Mid-parent heterosis


Critical thermal maximum




Funding information

This work was supported by grants from National Natural Science Foundation of China (No. 31472277, U1605213), Key S&T Program of Fujian & Shandong Province (No. 2016NZ01010006 and 2016GGH4513), and Earmarked Fund for Modern Agro-industry Technology Research System (No. CARS-49).


  1. Adema CM, Van der Knaap WPW, Siminia T (1991) Molluscan haemocyte-mediated cytotoxicity: the role of reactive oxygen intermediates. Rev Aquat Sci 4:201–223Google Scholar
  2. Babior BM (1978) Oxygen-dependent microbial killing by phagocytes (first of two parts). N Engl J Med 298:659–668CrossRefGoogle Scholar
  3. Bachère E, Hervio D, Miahle E (1991) Luminol-dependant chemiluminescence by haemocytes of the two marine bivalves, Ostrea edulis and Crassostrea gigas. Dis Aquat Org 11:173–180CrossRefGoogle Scholar
  4. Bansemer MS, Harris JO, Qin JG, Adams LR, Duong DN, Stone DA (2006) Growth and feed utilisation of juvenile greenlip abalone (Haliotis laevigata) in response to water temperatures and increasing dietary protein levels. Aquaculture 436:13–20CrossRefGoogle Scholar
  5. Bass DA, Parce JW, Dechatelet LR, Szejda P, Seeds MC, Thomas M (1983) Flow cytometric studies of oxidative product formation by neutrophils: a graded response to membrane stimulation. J Immunol 130:1910–1917Google Scholar
  6. Becker CD, Genoway RG (1979) Evaluation of the critical thermal maximum for determining thermal tolerance of freshwater fish. Environ Biol Fish 4(3):245–256CrossRefGoogle Scholar
  7. Buestel D, Ropert M, Prou J, Goulletquer P (2009) History, status, and future of oyster culture in France. J Shellfish Res 28(4):813–820CrossRefGoogle Scholar
  8. Cai W, Li S, Ma J (2004) Diseases resistance of Nile tilapia (Oreochromis niloticus), blue tilapia (Oreochromis aureus) and their hybrid (female Nile tilapia×male blue tilapia) to Aeromonas sobria. Aquaculture 229(1):79–87CrossRefGoogle Scholar
  9. Cai J, Han Y, Wang Z (2006) Isolation of Vibrio parahaemolyticus from abalone (Haliotis diversicolor supertexta L.) postlarvae associated with mass mortalities. Aquaculture 257(1):161–166CrossRefGoogle Scholar
  10. Cardinaud M, Offret C, Huchette S, Moraga D, Paillard C (2014) The impacts of handling and air exposure on immune parameters, gene expression, and susceptibility to vibriosis of European abalone Haliotis tuberculata. Fish Shellfish Immunol 36(1):1–8CrossRefGoogle Scholar
  11. Chang PH, Kuo ST, Lai SH, Yang HS, Ting YY, Hsu CL, Chen HC (2005) Herpes-like virus infection causing mortality of cultured abalone Haliotis diversicolor supertexta in Taiwan. Dis Aquat Org 65(1):23–27CrossRefGoogle Scholar
  12. Chang PH, Yang MC, Kuo ST, Chen MH, Cheng CH (2008) Occurrence of a rickettsia-like prokaryote in the small abalone, Haliotis diversicolor supertexta, cultured in Taiwan. Bull Eur Assoc Fish Pathol 28(2):52–57Google Scholar
  13. Chen M, Yang H, Delaporte M, Zhao SJ, Xing K (2007) Immune responses of the scallop Chlamys farreri after air exposure to different temperatures. J Exp Mar Biol Ecol 345(1):52–60CrossRefGoogle Scholar
  14. Cheng W, Hsiao IS, Hsu CH, Chen JC (2004a) Change in water temperature on the immune response of Taiwan abalone Haliotis diversicolor supertexta and its susceptibility to Vibrio parahaemolyticus. Fish Shellfish Immunol 17:235–243CrossRefGoogle Scholar
  15. Cheng W, Hsiao IS, Chen JC (2004b) Effect of ammonia on the immune response of Taiwan abalone Haliotis diversicolor supertexta and its susceptibility to Vibrio parahaemolyticus. Fish Shellfish Immunol 17(3):193–202CrossRefGoogle Scholar
  16. Cheng W, Juang FM, Chen JC (2004c) The immune response of Taiwan abalone Haliotis diversicolor supertexta and its susceptibility to Vibrio parahaemolyticus at different salinity levels. Fish Shellfish Immunol 16:295–306CrossRefGoogle Scholar
  17. Cowles RB, Bogert CM (1944) A preliminary study of the thermal requirements of desert reptiles. Bull Am Mus Nat Hist 83(5):261–296Google Scholar
  18. Deng Y, Liu X, Zhang G (2007) Fertilization, hatching, metamorphosis and growth oftwo Pacific abalone populations and their reciprocal crosses. Aquaculture 272:S319–S320Google Scholar
  19. Denny MW (1984) Mechanical properties of pedal mucus and their consequences forgastropod structure and performance. Am Zool 24(1):23–36CrossRefGoogle Scholar
  20. Denny MW, Gosline JM (1980) The physical properties of the pedal mucus of theterrestrial slug, Ariolimax columbianus. J Exp Biol 88(1):375–394Google Scholar
  21. Di G, Zhang Z, Ke C (2013) Phagocytosis and respiratory burst activity of haemocytes from the ivory snail, Babylonia areolata. Fish Shellfish Immunol 35(2):366–374CrossRefGoogle Scholar
  22. Di G, Kong X, Zhu G, Liu S, Zhang C, Ke C (2016) Pathology and physiology of Haliotis diversicolor, with withering syndrome. Aquaculture 453:1–9CrossRefGoogle Scholar
  23. Dı́az F, del Rı́o-Portı́lla MA, Sierra E, Aguilar M, Re-Araujo AD (2000) Preferred temperature and critical thermal maxima of red abalone Haliotis rufescens. J Therm Biol 25(3):257–261CrossRefGoogle Scholar
  24. Díaz F, Re AD, Medina Z, Re G, Valdez G, Valenzuela F (2006) Thermal preference and tolerance of green abalone Haliotis fulgens (Philippi, 1845) and pink abalone Haliotis corrugata (Gray, 1828). Aquac Res 37(9):877–884CrossRefGoogle Scholar
  25. Dweyer JJ, Burnett LE (1996) Acid-base status of the oyster Crassostrea virginica in response to air exposure and to infections by Perkinsus marinus. Biol Bull 190(1):139–147CrossRefGoogle Scholar
  26. Falconer DS, Mackay TFC (1996) Introduction to quantitative genetics, 4th edn. Longman Essex, EnglandGoogle Scholar
  27. Foley DA, Cheng TC (1975) A quantitative study of phagocytosis by hemolymph cells of the pelecypods Crassostrea virginica and Mecenaria mercenaria. J Invertebr Pathol 25:189–197CrossRefGoogle Scholar
  28. Ford T, Beitinger TL (2005) Temperature tolerance in the goldfish, Carassius auratus. J Therm Biol 30(2):147–152CrossRefGoogle Scholar
  29. Gilroy A, Edwards SJ (1998) Optimum temperature for growth of australian abalone: preferred temperature and critical thermal maximum for blacklip abalone Haliotis rubra (Leach), and greenlip abalone Haliotis laevigata (Leach). Aquac Res 29:481–485CrossRefGoogle Scholar
  30. Hahn KO (1989) Biotic and abiotic factors affecting the culture abalone. In: Hahn KO (ed) Handbook of culture of abalone and other marine gastropods. CRC Press, Boca RatonGoogle Scholar
  31. He L, Xin Z, Ying H, Yang H, Wang Y, Zhang Z (2017) The characterization of RHEB, gene and its responses to hypoxia and thermal stresses in the small abalone Haliotis diversicolor. Comp Biochem Physiol B Biochem Mol Biol 210:48–54CrossRefGoogle Scholar
  32. Hecht T (1994) Behavioural thermoregulation of the abalone Haliotis midae, and the implications for intensive culture. Aquaculture 26:171–181CrossRefGoogle Scholar
  33. Hégaret H, Wikfors GH, Soudant P (2003) Flow cytometric analysis of haemocytes from eastern oysters, Crassostrea virginica, subjected to a sudden temperature elevation: II. Haemocyte functions: aggregation, viability, phagocytosis, and respiratory burst. J Exp Mar Biol Ecol 293(2):249–265CrossRefGoogle Scholar
  34. Hooper C, Day R, Slocombe R, Handlinger J, Benkendorff K (2007) Stress and immune responses in abalone: limitations in current knowledge and investigative methods based on other models. Fish Shellfish Immunol 22(4):363–379CrossRefGoogle Scholar
  35. Hooper C, Day R, Slocombe R, Benkendorff K, Handlinger J, Goulias J (2014) Effects of severe heat stress on immune function, biochemistry and histopathology in farmed Australian abalone (hybrid Haliotis laevigata × Haliotis rubra). Aquaculture 432:26–37CrossRefGoogle Scholar
  36. Hsu TH, Gwo JC (2017) Genetic diversity and stock identification of small abalone (Haliotis diversicolor) in Taiwan and Japan. PLoS One 12(6):e0179818CrossRefGoogle Scholar
  37. Iwanaga S, Lee BL (2005) Recent advances in the innate immunity of invertebrate animals. BMB Rep 38(2):128–150CrossRefGoogle Scholar
  38. Jia X, Zhang Z, Wang S, Lin P, Zou ZH, Huang BQ, Wang YL (2009) Effects of tributyltin (TBT) on enzyme activity and oxidative stress in hepatopancreas and hemolymph of small abalone, Haliotis diversicolor supertexta. Chin J Oceanol Limnol 27:816–824CrossRefGoogle Scholar
  39. Knight JA (2000) Review: free radicals, antioxidants, and the immune system. Ann Clin Lab Sci 30:145–159Google Scholar
  40. Kumazawa NH, Morimoto N, Okamoto Y (1993) Luminol-dependent chemilumin-escence of haemocytes derived from marine and estuarine mollusks. J Vet Med Sci 55:287–290CrossRefGoogle Scholar
  41. Lafarga de la Cruz F, Gallardo-Escárate C (2011) Intraspecies and interspecies hybrids in Haliotis: natural and experimental evidence and its impact on abalone aquaculture. Rev Aquac 3:74–99CrossRefGoogle Scholar
  42. Lamkey KR, Edwards JW (1999) The quantitative genetics of heterosis. In: JG Coors and S Pandey (eds) Proceedings of the International Symposium on the Genetics and Exploitation of Heterosis in Crops, CIMMYT. Mexico City, Mexico. Aug. 17–22, 1997. pp 31–48Google Scholar
  43. Lee KK, Liu PC, Chen YC, Huang CY (2001) The implication of ambient temperature with outbreak of vibriosis in cultured small abalone Haliotis diversicolor supertexta Lischke. J Therm Biol 26:585–587CrossRefGoogle Scholar
  44. Leighton D, Lewis C (1983) Experimental hybridization in abalone. Invertebr Reprod Dev 5:273–282CrossRefGoogle Scholar
  45. Liang S, Luo X, You W, Luo X, Ke C (2014) The role of hybridization in improving the immune response and thermal tolerance of abalone. Fish Shellfish Immunol 39(1):69–77CrossRefGoogle Scholar
  46. Liang S, Luo X, You W, Luo X, Ke C (2018) Hybridization improved bacteria resistance in abalone: evidence from physiological and molecular responses. Fish Shellfish Immunol 72:679–689CrossRefGoogle Scholar
  47. Lin AYM, Brunner R, Chen PY, Talke FE, Meyers MA (2009) Underwater adhesion of abalone: the role of van der Waals and capillary forces. Acta Mater 57(14):4178–4185CrossRefGoogle Scholar
  48. Liu S, Mai K (2003) The progress of studies on molluscs immunological system and mechanism—a review. Acta Oceanol Sin 25:95–105Google Scholar
  49. Liu X, Yan Y, Wang Z, Cai M, Ke C (2008) A preliminary study on tolerance to high temperature and low salinity of Haliotis diversicolor Reeve. J Jimei Univ Natur Sci 4:301–303Google Scholar
  50. Livingstone DR (2001) Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms. Mar Pollut Bull 42:656–666CrossRefGoogle Scholar
  51. Lv ZM, Yang AG, Wang QY, Liu ZH, Zhou LQ (2006) Preliminary cytological identification and immunological traits of hybrid scallop from Chlamys farreri (♀) × Patinopecten yessoensis (♂). J Fish Sci China 13(4):597–602Google Scholar
  52. Marshall DJ, McQuaid CD (1993) Differential physiological and behavioural responses of the intertidal mussels, Choromytilus meridionalis (Kr.) and Perna perna L., to exposure to hypoxia and air: a basis for spatial separation. J Exp Mar Biol Ecol 171(2):225–237CrossRefGoogle Scholar
  53. Matozzo V, Ballarin L, Marin MG (2004) Exposure of the clam Tapes philippinarum to 4-nonylphenol: changes in anti-oxidant enzyme activities and re-burrowing capability. Mar Pollut Bull 48:563–571CrossRefGoogle Scholar
  54. Michaelidis B, Haas D, Grieshaber MK (2005) Extracellular and intracellular acid-base status with regard to the energy metabolism in the oyster Crassostrea gigas during exposure to air. Physiol Biochem Zool 78(3):373–383CrossRefGoogle Scholar
  55. Nie ZQ, Wang SP (2004) The status of abalone culture in China. J Shellfish Res 23(4):941–945Google Scholar
  56. Ospina AF, Mora C (2004) Effect of body size on reef fish tolerance to extreme low and high temperatures. Environ Biol Fish 70(4):339–343CrossRefGoogle Scholar
  57. Paladino FV, Spotila JR, Schubauer JP, Kowalski KT (1980) The critical thermal maximum: a technique used to elucidate physiological stress and adaptation in fishes. Rev Can Biol 39(2):115–122Google Scholar
  58. Park KI, Donaghy L, Kang HS, Hong HK, Kim YO, Choi KS (2012) Assessment of immune parameters of manila clam Ruditapes philippinarum in different physiological conditions using flow cytometry. Ocean Sci J 47(1):19–26CrossRefGoogle Scholar
  59. Pipe RK, Coles JA (1995) Environmental contaminants influencing immune function in marine bivalve molluscs. Fish Shellfish Immunol 5(8):581–595CrossRefGoogle Scholar
  60. Pipe RK, Coles JA, Thomas ME, Fossato VU, Pulsfor AL (1995) Evidence for environmentally derived immunomodulation in mussels from the Venice Lagoon. Aquat Toxicol 32(1):59–73CrossRefGoogle Scholar
  61. Rahman MF, Siddiqui MK (2004) Biochemical effects of vepacide (from Azadirachta indica) on Wistar rats during subchronic exposure. Ecotoxicol Environ Saf 59:332–339CrossRefGoogle Scholar
  62. Reynolds WW (1979) Perspective and introduction to the symposium: thermoregulation in ectotherms. Am Zool 19(1):193–194CrossRefGoogle Scholar
  63. Roch P (1999) Defense mechanisms and disease prevention in farmed marine invertebrates. Aquaculture 172(1):125–145CrossRefGoogle Scholar
  64. Rothe G, Valet G (1990) Flow cytometric analysis of respiratory burst activity in phagocytes with hydroethidine and 2′, 7′-dichlorofluorescin. J Leukocyte Boil 47(5):440–448CrossRefGoogle Scholar
  65. Soares-da-Silva IM, Ribeiro J, Valongo C, Pinto R, Vilanova M, Bleher R, Machado J (2002) Cytometric, morphologic and enzymatic characterisation of haemocytes in Anodonta cygnea. Comp Biochem Phys A 132(3):541–553CrossRefGoogle Scholar
  66. Song ZR, Ji RX, Yan SF, Chen CS, Zhong YP, Jiang YH, Ni ZM (2000) A spherovirus resulted in mass mortality of Haliotis diversicolor aquatilis. J Fisheries China 24(5):463–467Google Scholar
  67. Wang SH, Wang YL, Zhang ZX (2004) Different response of innate immune factors in abalone Haliotis diversicolor supertexta to E. coli or Vibrio parahaemolyticus infection. J Shellfish Res 23:1173–1177Google Scholar
  68. Wells RMG, Baldwin J (1995) A comparison of metabolic stress during air exposure in two species of New Zealand abalone, Haliotis iris and Haliotis australis: implications for the handling and shipping of live animals. Aquaculture 134(3):361–370CrossRefGoogle Scholar
  69. Xue QG, Renault T, Chilmonczyk S (2001) Flow cytometric assessment of haemocyte sub-populations in the European flat oyster, Ostrea edulis, haemolymph. Fish Shellfish Immunol 11:557–567CrossRefGoogle Scholar
  70. Yakovleva NV, Samoilovich MP, Gorbushin AM (2001) The diversity of strategies of defense from pathogens in molluscs. J Evol Biochem Physiol 37:358–367CrossRefGoogle Scholar
  71. Yang C, Kong J, Wang Q, Liu QH, Tian Y, Luo K (2007) Heterosis of haemolymph analytes of two geographic populations in Chinese shrimp Fenneropenaeus chinensis. Fish Shellfish Immunol 23(1):62–70CrossRefGoogle Scholar
  72. You WW, Ke CH, Luo X, Wang DX (2009) Growth and survival of three small abalone Haliotis diversicolor populations and their reciprocal crosses. Aquac Res 40(13):1474–1480CrossRefGoogle Scholar
  73. You WW, Luo X, Wang DX, Lin ZB, Lin HY, Ke CH (2010) Comparisons of morphological characteristics and grow-out performance in new variety Dongyou No. 1 and its parental populations of small abalone Haliotis diversicolor. J Fisheries China 12:1837–1843Google Scholar
  74. You WW, Lin HY, Luo X, Wang DX, Lin ZB, Ke CH (2011a) Effects of temperature on the growth rates and survival rates of Haliotis diversicolor among different populations. J Oceanography Taiwan Strait 30(4):583–588Google Scholar
  75. You WW, Zhan X, Wang DX, Li WD, Luo X, Ke CH (2011b) Genetic variation analysis in wild and cultured subpopulations of small abalone Haliotis diversicolor estimated by microsatellite markers. N Am J Aquac 73(4):445–450CrossRefGoogle Scholar
  76. Yu RH, Wang ZP, Kong LF, Li Q, Zheng XD (2006) A study on the survival rate of Pacific oysters in different exposure states at different development stages. J Ocean Univ China 36(4):617–620Google Scholar
  77. Zhang GF, Que HY, Liu X, Xu HS (2004) Abalone mariculture in China. J Shellfish Res 23(4):947–950Google Scholar
  78. Zheng HP, Zhang GF, Guo XM, Liu X (2006) Heterosis between two stocks of the bay scallop, Argopecten irradians irradians Lamarck. J Shellfish Res 25:807–812CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2018

Authors and Affiliations

  • Weiwei You
    • 1
    • 2
    • 3
    Email author
  • Bo Wang
    • 1
    • 4
  • Xuan Luo
    • 1
    • 3
  • Caihuan Ke
    • 1
    • 2
    • 3
  1. 1.College of Ocean and Earth ScienceXiamen UniversityXiamenChina
  2. 2.State Key Laboratory of Marine Environmental ScienceXiamen UniversityXiamenChina
  3. 3.Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological ResourcesXiamenPeople’s Republic of China
  4. 4.Yantai Marine Property Rights Exchange CenterYantaiChina

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