Skip to main content

Advertisement

SpringerLink
Log in

Comparative effects of Aloe vera gel and Freund’s adjuvant in vaccination of common carp (Cyprinus carpio L.) against Aeromonas hydrophila

  • Published:
Aquaculture International Aims and scope Submit manuscript

Abstract

Natural compounds of Aloe vera gel are well known for their immunostimulatory properties. In this study, adjuvant effects of Aloe vera in combination with formalin-killed A. hydrophila bacterin in immunization of common carp were evaluated and compared with Freund’s adjuvant. A total of 480 juvenile carp were randomly divided into four groups. Groups 1–4 were injected on days 0 and 14 with normal saline (as control), bacterin, bacterin + Freund’s adjuvant and bacterin + Aloe vera gel, respectively. Blood samples were taken on days 14, 28 and 42. Hematoimmunological parameters including white blood cell (WBC) count, alternative complement activity, serum lysozyme and bactericidal activity, respiratory burst activity (NBT) and antibody titer were evaluated and compared among different groups. At the end of experiment, all groups were challenged with live A. hydrophila and relative percentage survival (RPS) compared among groups. According to the results, administration of both adjuvants caused a positive change in fish immune response and most of the aforementioned parameters significantly improved in vaccinated groups compare with control group (P < 0.05). Moreover, WBC count, alternative complement activity, serum lysozyme activity and NBT activity were significantly higher in Aloe adjuvant group compared with Freund’s adjuvant (P < 0.05). Finally after challenge, groups 3 and 4 showed significantly lesser mortality than groups 1 and 2. Overall, our results indicated that combination of A. hydrophila bacterin with adjuvant can improve the vaccine efficacy and resistance against A. hydrophila infection, and in comparison with traditional adjuvants, Aloe vera gel could be used as a natural adjuvant with similar or even greater positive effects on vaccination of common carp.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Abdy E, Alishahi M, Tabandeh MR, Ghorbanpoor M, Jafari H (2016) Comparative effect of Freund’s adjuvant and Aloe vera L. gel on the expression of TNF-α and IL-1β in vaccinated Cyprinus carpio L. with Aeromonas hydrophila C. bacterin. Aquac Res 47:2543–2552

    Article  CAS  Google Scholar 

  • Alexander CP, Kirubakaran CJ, Michael RD (2010) Water soluble fraction of Tinospora cordifolia leaves enhanced the non-specific. Fish Shellfish Immunol 29:765–772

    Article  PubMed  Google Scholar 

  • Alishahi M, Abdy E (2013) Effects of different levels of Aloe vera L. extract on growth performance, hemato-immunological indices of Cyprinus carpio L. Iran J Vet Sci Technol 5(2):33–44

    Google Scholar 

  • Alishahi M, Ranjbar MM, Ghorbanpour M, Peyghan R, Mesbah M, Razijalali M (2010) Effects of dietary Aloe vera on specific and nonspecific immunity of common carp (Cyprinus carpio). Iran J Vet Res 4(3):85–91

    Google Scholar 

  • Amend DF (1981) Potency testing of fish vaccines. In: Anderson DP, Hennessen H (eds) Fish biologies: serodiagnostics and vaccines. Development in biological standardization, vol 49. Karger, Basel, pp 447–454

    Google Scholar 

  • Anderson DP, Siwicki AK (1994) Duration of protection against Aeromonas salmonicida in brook trout immuno stimulated with glucan or chitosan by injection and immersion. Prog Fish Cultur 56:258–261

    Article  Google Scholar 

  • Anderson DP, Moritomo T, de Grooth R (1992) Neutrophil, glass-adherent, nitroblue tetrazolium assay gives early indication of immunization effectiveness in rainbow trout. Vet Immunol Immunopathol 30(4):419–429

    Article  CAS  PubMed  Google Scholar 

  • Austin B, Austin DA (1989) Microbiological examination of fish and shellfish. Ellis Horwood, Chichester, pp 217–488

    Google Scholar 

  • Azza MM (2009) Antagonism of Aeromonas hydrophila by propolis and its effect on the performance of Nile tilapia, Oreochromis niloticus. Fish Shellfish Immunol 27:454–459

    Article  Google Scholar 

  • Bastardo A, Ravelo C, Castro N, Calheiros J, Romalde L (2012) Effectiveness of bivalent vaccines against Aeromonas hydrophila and Lactococcus garvieae infections in rainbow trout Oncorhynchus mykiss (Walbaum). Fish Shellfish Immunol 32(5):756–761

    Article  CAS  PubMed  Google Scholar 

  • Carter SJ (2008) Cooper and Gunn’s dispensing pharmacy, 12th edn. CBS publisher, New Delhi, pp 541–572

    Google Scholar 

  • Chinnah AD, Baig MA, Tizard IR, Kemp MC (1992) Antigen dependent adjuvant activity of a polydispersed beta-(1,4)-linked acetylated mannan (acemannan). Vaccine 10:551–557

    Article  CAS  PubMed  Google Scholar 

  • Chu WH (2006) Adjuvant effect of propolis on immunisation by inactivated Aeromonas hydrophila in carp Carassius auratus gibelio. Fish Shellfish Immunol 21:113–117

    Article  CAS  PubMed  Google Scholar 

  • Costa A, Leef M, Bridle AR, Carson J, Nowak BF (2011) Effect of vaccination against yersiniosis on the relative percent survival, bactericidal and lysozyme response of Atlantic salmon, Salmo salar. Aquaculture 315:201–206

    Article  CAS  Google Scholar 

  • Divyagnaneswari M, Christybapita D, Michael RD (2007) Enhancement of nonspecific immunity and disease resistance in Oreochromis mossambicus by Solanum trilobatum leaf fractions. Fish Shellfish Immunol 23:249–259

    Article  CAS  PubMed  Google Scholar 

  • Djeraba A, Quere P (2000) In vivo macrophage activation in chickens with acemannan, a complex carbohydrate extracted from Aloe vera. Int J Immunopharmacol 22:365–372

    Article  CAS  PubMed  Google Scholar 

  • Dotta G, de Andrade JI, Tavares Gonçalves EL, Brum A, Mattos JJ, Maraschin M, Martins ML (2014) Leukocyte phagocytosis and lysozyme activity in Nile tilapia fed supplemented diet with natural extracts of propolis and Aloe barbadensis. Fish Shellfish Immunol 39:280–284

    Article  CAS  PubMed  Google Scholar 

  • Ebanks RO, Dacanay A, Goguen M, Pinto DM, Ross NW (2004) Differential proteomic analysis of Aeromonas salmonicida outer membrane proteins in response to low iron and in vivo growth conditions. Proteomics 4:1074–1085

    Article  CAS  PubMed  Google Scholar 

  • Egger S, Brown GS, Kelsey LS, Yates KM, Rosenberg LJ, Talmadge JE (1996) Studies on optimal dose and administration schedule of a hematopoietic stimulatory (1,4)-linked mannan. Int J Immunopharmacol 18:113–126

    Article  CAS  PubMed  Google Scholar 

  • Ellis AE (1990) Lysozyme assays. In: Stolen JS, Fletcher TC, Anderson DP, Roberson BS, van Muiswinkel WB (eds) Techniques in fish immunology. SOS Publications, New Jersey, pp 101–113

    Google Scholar 

  • Evensen O, Brudeseth BE, Mutoloki S (2005) The vaccine formulation and its role in inflammatory processes in fish effects and adverse effects. Dev Biol Stand 121:117–125

    CAS  Google Scholar 

  • Fang HM, Ge R, Sin YM (2004) Cloning, characterisation and expression of Aeromonas hydrophila major adhesin. Fish Shellfish Immunol 16:645–658

    Article  CAS  PubMed  Google Scholar 

  • Freund J (1956) The mode of action of immunological adjuvants. Adv Tuber Res 7:130–148

    Google Scholar 

  • Gabriel NN, Qiang J, He J, Ma XY, Kpundeh MD, Xu P (2015) Dietary Aloe vera supplementation on growth performance, some haemato-biochemical parameters and disease resistance against Streptococcus iniae in tilapia (GIFT). Fish Shellfish Immunol 44:504–514

    Article  CAS  PubMed  Google Scholar 

  • Gjessing MC, Falk K, Weli SC, Koppang EO, Kvellestad A (2012) A sequential study of incomplete Freund’s adjuvant-induced peritonitis in Atlantic cod. Fish Shellfish Immunol 32:141–150

    Article  CAS  PubMed  Google Scholar 

  • Green P (1996) Aloe vera extracts in equine clinical practice. Vet Times 26:16–18

    Google Scholar 

  • Gudding R, Lillehaug A, Evensen O (1999) Recent developments in fish vaccinology. Vet Immunol Immunopathol 72:203–212

    Article  CAS  PubMed  Google Scholar 

  • Gupta RK, Rost BE, Relyveld E, Siber GR (1995) Adjuvant properties of aluminium and calcium compounds. In: Powell MF, Newman MJ (eds) Vaccine design: the subunit and adjuvant approach. Plenum Press, New York, pp 229–248

    Chapter  Google Scholar 

  • Habeeb F, Stables G, Bradbury F, Nong S (2007) The inner gel component of Aloe vera suppresses bacterial-induced pro-inflammatory cytokines from human immune cells. Methods 42:388–393

    Article  CAS  PubMed  Google Scholar 

  • Harikrishnan R, Balasundaram C, Heo MS (2011) Impact of plant products on innate and adaptive immune system of cultured finfish and shellfish. Aquaculture 317:1–15

    Article  CAS  Google Scholar 

  • Harikrishnan R, Kim JS, Balasundaram C, Heo MS (2012) Vaccination effect of liposomes entrapped whole cell bacterial vaccine on immune response and disease protection in Epinephelus bruneus against Vibrio harveyi. Aquaculture 342:69–74

    Article  Google Scholar 

  • Jang SI, Marsden MJ, Kim YG, Choi MS, Secombes CJ (1995) The effect of glycyrrhizin on rainbow trout, Oncorhynchus mykiss, leucocyte responses. J Fish Dis 18:307–315

    Article  CAS  Google Scholar 

  • Jiao XD, Cheng S, Hu YH, Sun L (2010) Comparative study of the effects of aluminum adjuvants and Freund’s incomplete adjuvant on the immune response to an Edwardsiella tarda major antigen. Vaccine 28:1832–1837

    Article  CAS  PubMed  Google Scholar 

  • Kamilya D, Maiti T, Joardar S, Mal B (2006) Adjuvant effect of mushroom glucan and bovine lactoferrin upon Aeromonas hydrophila vaccination in catla, Catla catla (Hamilton). J Fish Dis 29:331–337

    Article  CAS  PubMed  Google Scholar 

  • Kim KH, Hwang YJ, Bai S (1999) Resistance to Vibrio alginolyticus in juvenile rockfish (Sebastes schlegeli) fed diets containing different doses of aloe. Aquaculture 180:13–21

    Article  Google Scholar 

  • Kwon SR, Nam YK, Kim SK, Kim KH (2006) Protection of tilapia (Oreochromis mosambicus) from edwardsiellosis by vaccination with Edwardsiella tarda ghosts. Fish Shellfish Immunol 20(4):621–626

    Article  CAS  PubMed  Google Scholar 

  • Lee YC (1988) Mannose-binding proteins of animal origin. In: The molecular immunology of complex carbohydrates. Springer, US, pp 105–121

    Chapter  Google Scholar 

  • Lee CK (2006) Efficacy of aloe. In: New perspectives on aloe. Springer, Berlin, pp 63–167

  • Lee JK, Lee MK, Yun YP, Kim Y, Kim JS (2001) Acemannan purified from Aloe vera induces phenotypic and functional maturation of immature dendritic cells. Int J Immunopharmacol 1:1275–1284

    Article  CAS  Google Scholar 

  • Lorenzen N, Cupit PM, Einer-Jensen K, Lorenzen E, Ahrens P, Secombes CJ (2000) Protection against a viral disease in fish following injection of plasmid DNA encoding neutralising single chain antibody. Nat Biotechnol 18:1177–1180

    Article  CAS  PubMed  Google Scholar 

  • Magnadottir B (2006) Innate immunity of fish, overview. Fish Shellfish Immunol 20:137–151

    Article  CAS  PubMed  Google Scholar 

  • Misra CK, Das BK, Mukherjee SC, Meher PK (2006a) The immunomodulatory effects of tuftsin on the non-specific immune system of Indian Major carp, Labeo rohita. Fish Shellfish Immunol 20:728–738

    Article  CAS  PubMed  Google Scholar 

  • Misra CK, Das BK, Mukherjee SC, Pattnaik P (2006b) Effect of multiple injections of beta-glucan on non-specific immune response and disease resistance in Labeo rohita fingerlings. Fish Shellfish Immunol 20(3):305–319

    Article  CAS  PubMed  Google Scholar 

  • Nordgren RM, Stewart-Brown B, Rodeberg JH (1992) The role of acemannan as an adjuvant for Marek’s disease vaccine. In: Proceedings of the XIX worlds poultry congress, Amsterdam, The Netherlands

  • Ortuno J, Esteban MA, Mulero V, Meseguer J (1998) Methods for studying the haemolytic, chemoattractant and opsornic activities of seabream serum. In: Barnes AC, Davidson GA, Hiney MP, Mclntosh D (eds) Methodology in fish diseases research, vol I. Fisheries Research Services, Aberdeen, pp 97–100

    Google Scholar 

  • Peddie S, Zou J, Secombes CJ (2002) Immunostimulation in the rainbow trout (Oncorhynchus mykiss) following intraperitoneal administration of Ergosan. Vet Immunol Immunopathol 86:101–113

    Article  CAS  PubMed  Google Scholar 

  • Peng SY, Norman J, Curtin G, Corrier D, McDaniel HR, Busbee D (1991) Decreased mortality of Norman murine sarcoma in mice treated with the immunomodulator, acemannan. Mol Biother 3:79–87

    CAS  PubMed  Google Scholar 

  • Perez O, Romeu B, Cabrera O, González E, Batista-Duharte A, Labrada A, Pérez R, Reyes L, Ramírez W, Sifontes S, Fernández N, Lastre M (2013) Adjuvants are key factors for the development of future vaccines: lessons from the finlay adjuvant platform. Front Immunol 4:407

    Article  PubMed  PubMed Central  Google Scholar 

  • Petrovsky N, Aguilar JC (2004) Vaccine adjuvants: current state and future trends. Immunol Cell Biol 82:488–496

    Article  CAS  PubMed  Google Scholar 

  • Raa J (1996) The use of immuno-stimulatory substances in fish and shellfish farming. Rev Fish Sci 4:229–288

    Article  Google Scholar 

  • Reed LV, Muench H (1938) A simple method of estimating fifty percent end points. Am J Hyg 27:493–497

    Google Scholar 

  • Reynolds T, Dweck AC (1999) Aloe vera leaf gel: a review update. J Ethnopharmacol 68:3–37

    Article  CAS  PubMed  Google Scholar 

  • Sakai M (1999) Current research status of fish immunostimulants. Aquaculture 172:63–92

    Article  CAS  Google Scholar 

  • Saurabh S, Sahoo PK (2008) Lysozyme: an important defence molecule of fish innate immune system. Aquac Res 39:223–239

    Article  CAS  Google Scholar 

  • Schaperclaus W, Kulow H, Schreckenbach K (1991) Hematological and serological technique. In: Kothekar VS (ed) Connaught circus, vol 1, 2nd edn. Gulab primlani, Oxonian press Pvt. Ltd, New Delhi, pp 71–108

    Google Scholar 

  • Soltani M, Kalbassi MR (2001) Protection of Persian sturgeon (Acipenser persicus) fingerling against Aeromonas hydrophila septicemia using three different antigens. Bull Eur Assoc Fish Pathol 21(6):235

    Google Scholar 

  • Stuart RW, Lefkowitz DL, Lincoln JA, Howard K, Gelderman MP, Lefkowitz SS (1997) Upregulation of phagocytosis and candidicidal activity of macrophages exposed to the immunostimulant, acemannan. Int J Immunopharmacol 19:75–82

    Article  CAS  PubMed  Google Scholar 

  • Sun Y, Liu CS, Sun L (2011a) A multivalent killed whole-cell vaccine induces effective protection against Edwardsiella tarda and Vibrio anguillarum. Fish Shellfish Immunol 31:595–599

    Article  CAS  PubMed  Google Scholar 

  • Sun Z, Wei K, Yan Z, Zhu X, Wang X (2011b) Effect of immunological enhancement of aloe polysaccharide on chickens immunized with Bordetella avium inactivated vaccine. Carbohydr Polym 86:684–690

    Article  CAS  Google Scholar 

  • Swain P, Dash S, Sahoo PK, Routray P, Sahoo SK, Gupta SD, Meher PK, Sarangi N (2006) Non-specific immune parameters of brood Indian major carp Labeo rohita and their seasonal variations. Fish Shellfish Immunol 22:38–43

    Article  PubMed  Google Scholar 

  • Tafalla C, Bogwald J, Dalmo RA (2013) Adjuvants and immunostimulants in fish vaccines: current knowledge and future perspectives. Fish Shellfish Immunol 35:1740–1750

    Article  CAS  PubMed  Google Scholar 

  • Talmadge J, Chavez J, Jacobs L, Munger C, Chinnah T, Chow JT, Williamson D, Yates K (2004) Fractionation of Aloe vera L. inner gel, purification and molecular profiling of activity. Int Immunopharmacol 4:1757–1773

    Article  CAS  PubMed  Google Scholar 

  • Wang N, Yang Z, Zang M, Liu Y, Lu C (2013) Identification of Omp38 by immunoproteomic analysis and evaluation as a potential vaccine antigen against Aeromonas hydrophila in Chinese breams. Fish Shellfish Immunol 34:74–81

    Article  PubMed  Google Scholar 

  • Wanman CH, Tanmark N, Supamattaya K (2007) Production of killed vaccine from Streptococcus sp. and its application in sea bass (Lates calcarifer). Songklanakarin J Sci Technol 29:1251–1261

    Google Scholar 

  • Wonbel D, Helderman H (1988) Enhancement of allo responsiveness of human lymphocytes by acemannan (carrisyn2). Int J Immunopharmacol 10:967–976

    Article  Google Scholar 

  • Wu N, La-Patra SE, Li J, Sunyer JO, Zhang YA (2014) Complement C5a acts as molecular adjuvant in fish by enhancing antibody response to soluble antigen. Fish Shellfish Immunol 40:616–623

    Article  CAS  PubMed  Google Scholar 

  • Xia C, Ma ZH, Rahman MH, Wu ZG (2004) PCR cloning and identification of the β-haemolysin gene of Aeromonas hydrophila from freshwater fishes in China. Aquaculture 229:45–53

    Article  CAS  Google Scholar 

  • Yates KM, Rosenberg LJ, Harris CK, Bronstad DC, King GK, Biehle GA, Walker B, Ford CR, Hall JE, Tizard IR (1992) Pilot study of the effect of acemannan in cats infected with feline immunodeficiency virus. Vet Immunol Immunopathol 35:177–189

    Article  CAS  PubMed  Google Scholar 

  • Yin G, Ardo L, Thompson K, Adams A, Jeney Z, Jeney G (2009) Chinese herbs (Astragalus radix and Ganoderma lucidum) enhance immune response of carp, Cyprinus carpio, and protection against Aeromonas hydrophila. Fish Shellfish Immunol 26:140–145

    Article  PubMed  Google Scholar 

  • Zanuzzo FS, Urbinati EC, Nash GW, Gamperl AK (2015a) Steelhead trout Oncorhynchus mykiss metabolic rate is affected by dietary Aloe vera inclusion but not by mounting an immune response against formalin-killed Aeromonas salmonicida. J Fish Biol 87(1):43–53

    Article  CAS  PubMed  Google Scholar 

  • Zanuzzo FS, Urbinati EC, Rise ML, Hall JR, Nash GW, Gamperl AK (2015b) Aeromonas salmonicida induced immune gene expression in Aloe vera fed steelhead trout, Oncorhynchus mykiss (Walbaum). Aquaculture 435:1–9

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was funded by a Grant from Shahid Chamran University of Ahvaz research Council. We would like to thank Prof. Soltani (Department of Aquatic Animal Health, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran) for providing us with Aeromonas hydrophila AH-04.

Author information

Author notes

  1. Esmail Abdy

    Present address: Department of Aquatic Sciences, Faculty of Veterinary Medicine, Tehran University, Tehran, Iran

Authors and Affiliations

  1. Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Khuzestan, 61357-831351, Iran

    Esmail Abdy, Mojtaba Alishahi & Takavar Mohammadian

  2. Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

    Masoud Ghorbanpour

  3. Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

    Morteza Tollabi

Authors
  1. Esmail Abdy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mojtaba Alishahi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Morteza Tollabi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Masoud Ghorbanpour
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Takavar Mohammadian
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Esmail Abdy.

Ethics declarations

Conflict of interest

The authors declare that there is no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Abdy, E., Alishahi, M., Tollabi, M. et al. Comparative effects of Aloe vera gel and Freund’s adjuvant in vaccination of common carp (Cyprinus carpio L.) against Aeromonas hydrophila . Aquacult Int 25, 727–742 (2017). https://doi.org/10.1007/s10499-016-0074-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10499-016-0074-1

Keywords

Search

Navigation