The effects of anti-Streptococcosis vaccine by bath method on the levels of cortisol and lactate dehydrogenase (LDH) and the expression of immune-related genes in Asian seabass (Lates calcarifer)

Document Type : (original research)

Authors

1 Department of Biology, Faculty of Biological Sciences, Varamin-Pishva Branch, Islamic Azad University, Pishva, Iran

2 Department of Fisheries, Persian Gulf Research Institute, Persian Gulf University, Bushehr, Iran

3 Iranian Fisheries Science Research Institute, Agriculture Research Education and Organization , Tehran, Iran

4 Faculty of Veterinary Medicine, Science and Research Unit, Islamic Azad University, Tehran, Iran

5 Department of Biology, Faculty of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran

10.22034/aej.2024.453286.3123

Abstract

The aim of this study was to investigate the effects of immersion vaccine (Garovak) against streptococcosis and lactococcosis in different weights on the growth, feeding and survival rates of Asian seabass (Lates calcarifer). Fish with an average weight of 5 gr were studied in 300-liter round polyethylene tanks in 2 treatments and 3 repetitions, including the vaccinated treatments at a weight of 5 grams (ST) and the control group without vaccination. Feeding was done twice a day until satiety for 8 weeks. The results related to the bacterial challenge showed that the death rate in the vaccinated treatment was significantly lower than the control group (P<0.05). According to the general results of this study, the administration of Garovak immersion vaccine has been able to improve the survival rate after bacterial challenge in Asian seabass weighing 5 grams. In the statistical analysis of the results of the present study using SPSS software tests, the expression of genes related to immunity using Real-time (RT)-PCR, which has a significant difference in the gene expression of treatment samples was observed with control samples. Also, the average levels of cortisol and lactate dehydrogenase (LDH) enzymes in the tested treatments were 107 ng/g and 1279 u/kg, which had no significant difference with the control group. Also, in the statistical analysis, no significant difference was observed between gene expression data (Lysozymes, TNF, MHCI, and IGF-I). But in 1L-IB and MHCII, was seen significant difference between treatments and controls. However, the average gene expression (Lysozymes, 1L-IB, TNF, MHCI, MHCII and IGF-I) in the vaccinated treatment was higher than the average values of the control treatment. The results obtained in the current study showed that the vaccine had a better effect on the health of Asian sea bass weighing 5 gr. According to the general results of this study, the vaccine has been able to have a positive effect on fish health indicators, but on cortisol and lactate dehydrogenase enzymes (LDH) and also gene expression (Lysozymes, TNF, MHCI, and IGF-I) had no significant effect. The results obtained in the current study showed that the vaccine had a better effect on the health of Asian sea bass weighing 5 grams. But it had no significant effect on the levels of cortisol and lactate dehydrogenase (LDH). While it had a significant effect on MHC1 and interleukin 1B gene expression.

Keywords

Main Subjects


  1. FAO. 2022. State of World Fisheries and Aquaculture (SOFIA), Food and Agricultural Organization of the United Nations, Rome.
  2. Azhdahakoshpour, A., Payghan, R. and Ahangarzadeh, M., 2017. Most important diseases of cage cultured marine fish with emphasis on Asian sea bass (lates calcarifer) and prevention methods. Journal of Marine Fishes. 1(1): 11-22. (In Persian)
  3. Garza-Gil, M.D., Varela-Lafuente, M. and Caballero-Miguez, G., 2009. Price and production trends in the marine fish aquaculture in Spain. Aquaculture Research. 40: 274-281.
  4. Hajirezaee, S., Ajdar, D., Matinfar, A., Hosseini Aghuzbeni, S. and Rafiee, G., 2015. A 1earthen ponds of Gwadar region, Iran: an assessment of growth parameters feed intake efficiency and survival rate. Journal of Applied Animal Research. 43(3): 309-313. https://doi.org/10.1080/ 09712119.2014.963105
  5. FAO. 2024. Globefish Highlights International markets for fisheries and aquaculture products. Food and Agricultural Organization of the United Nations, Rome.
  6. Gibson‐Kueh, S., Chee, D., Chen, J., Wang, Y.H., Tay, S., Leong, L.N., Ng, M.L., Jones, J.B., Nicholls, P.K. and Ferguson, H.W., 2012. The pathology of ‘scale drop syndrome in Asian seabass, Lates calcarifer Bloch, a first description. Journal of fish diseases. 35(1): 19-27. https://doi.org/10.1111/j.1365-2761.2011.01319.x
  7. Bromage, E.S., Thomas, A. and Owens, L., 1999. Streptococcus iniae, a bacterial infection in barramundi Lates calcarifer. Diseases of aquatic organisms. 36(3): 177-181. doi:10.3354/dao036177
  8. Harikrishnan, R., Balasundaram, C. and Heo, M.S., 2012. Poly D, L-lactide-co-glycolic acid (PLGA)- encapsulated vaccine on immune system in Epinephelus bruneus against Uronema marinum. Experimental Parasitology. 131: 325-332. https://doi.org/10.1016/j. exppara.2012.04.0176
  9. Raa, J., 1992. The use of immunostimulants to increase resistance of aquatic organism to microbial infections. Diseases in Asian aquaculture. 39-50. https://doi.org/10. 11233/aquaculturesci.57.167
  10. Mazandarani, M., Hoseinifar, S.H., Reza Gholitebar, Z., Soudagar, M. and Safari, R., 2022. Evaluation of Yersinia ruckeri vaccine performance in rainbow trout (Oncorhynchus mykiss). Journal of Utilization and Cultivation of Aquatics. 11(2): 37-48. (In Persian)
  11. Mohammadi, Y., Mesbah, M., Dezfoulnejad, M.C., Mehrgan, M.S. and Islami, H.R., 2021. Growth performance, blood biochemical parameters, immune response, and antioxidant defense of Asian seabass (Lates calcarifer) fingerlings exposed to monovalent and bivalent vaccines against Streptococcus iniae and Vibrio harveyi. Aquaculture International. 29:2751-2767. https://doi.org/10.1007/s 10499-021-00776-5.
  12. Alishahi, M., Halimi, M., Ghorbanpoor Najaf abadi, M. and Erfanmanesh, A., 2020. Comparison of efficacy and immunogenicity of three administration routs of Streptococosis/lactococosis vaccine in reinbow trout. Journal of Animal Environment. 11(4): 187-196. (In Persian)
  13. Khaj, H., Mesbah, M., Tabandeh, M.R., Mohammadian, T. and Dadar, M., 2018. Comparative effects of Iranian streptococcus/lactococcus vaccine and Aquavac vaccine on growth performance and immunity in rainbow trout (Onchorhynchus mykiss). Iranian Veterinary Journal. 13(4): 28-42. (In Persian)
  14. Poursaeid, S., Falahatkar, B., Mojazi Amiri, B. and Van Der Kraak, G., 2012. Effects of long -term cortisol treatments on gonadal development, sex steroids levels and ovarian cortisol content in cultured great sturgeon (Huso huso). Comparative Biochemistry and Physiology. 163: 111-119.
  15. Thomas, P., Woodin, B.R. and Neff, J.M., 1980. Biochemical responses of the striped mullet Mugil cephalus to oil exposure: I. Acute responses-interrenal activations and secondary stress responses. Mar. Biol. 59: 114-141.
  16. Sambrook, J. and Russell, D.W., 2001. Molecular Cloning: A Laboratory Manual. 3rd Edition, Vol. 1, Cold Spring Harbor Laboratory Press, New York.
  17. Livak, K.J. and Schmittgen, T.D., 2001. Analysis of Relative Gene Expression Data Using Real Time Quantitative PCR and the 22DDCT Method. Methods. 25: 402-408.
  18. Pfaffl, M.W., 2001. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 1: 29(9): e45. doi: 10.1093/nar/29.9.e45. PMID: 11328886; PMCID: PMC55695.
  19. Vendrell, D., Balcázar, J.L., Ruiz-Zarzuela, I., de Blas, I., Gironés, O. and Múzquiz, J.L., 2006. Lactococcus garvieae in fish: a review. Comp Immunol Microbiol Infect Dis. 29(4): 177-198. doi: 10.1016/j.cimid.2006.06.003. PMID: 16935332.
  20. Gudding, R., Lillehaug, A. and Evensen, Ø., 1999. Recent developments in fish vaccinology. Vet Immunol Immunopathol. 72(1-2): 203-212.
  21. Hastein, T., Scarfe, A.D. and Lund, V.L., 2005. Science-based assessment of welfare: Aquatic animals. Rev. Sci. Tech. 24(2): 529.
  22. Otatake, M., Kiryu, I. and Nakanishi, T., 2002. Development of vaccine delivery method for fish: Parcutaneous administration by immersion with application of multiple puncture instrument. Journal of Vaccine. 1: 3764-3769.
  23. Sakai, T., Sakamoto, T., Hallaert, J. and Vandamme, E.J., 1993. Pectin, pectinase and protopectinase: production, properties, and applications. Adv Appl Microbiol. 39: 213-294. doi: 10.1016/s0065-2164(08)70597-5. PMID: 8213306.
  24. Bayunova, L., Barannikova, I. and Semenkova, T., 2002. Sturgeon stress reactions in aquaculture. Journal of Applied Ichthyology. 18(4-6): 397-404.
  25. Morshedi, V., Bakhshi, N., Ebrahimi, H. and Yousefi Siahkalroudi, S., 2023. Effects of using GaroVak immersion vaccine in rearing the juvenile Asian sea bass (Lates calcarifer) and experimentally infected with Streptococcus iniae. Journal of Animal Environment. 15(1): 191-200. (In Persian)
  26. Akrami, R., Ahmadi, Z., Chitsaz, H., Shamloofar, M., Habibi Nodeh, F., Sadeghi Asl, F. and Zarrini, N., 2018. Effect of feed supplemented with ginger (Zingiber officinale) extract on the growth, biochemical and hemato-immunological parameters of rainbow trout (Oncorhynchus mykiss). Journal of Veterinary Research. 73(2): 155-163. (In Persian)
  27. Makvandi, H., Khodadadi, M., Keyvan Shokoh, S. and Makvandi, Z., 2012. Effect of salinity stress on cortisol hormone and glucose in grass carp fingerlings (Ctenopharyngodon idella). Journal of Aquatic Animals and Fisheries. 2(8): 77-84.
  28. Lim, C., Yildirim-Aksoy, M. and Welker, T., Effect of feeding duration of sodium chloride containing diets on growth performance and some osmoregulatory parameters of Nile tilapia (Oreochromis niloticus) after transfer to water of different salinities. In: burright, j., Flemming, C. and Egna, H., (eds). Twenty-Second Annual Technical Report. Aquaculture CRSP, Oregon State University, Corvallis, Oregon. 411-420.
  29. Navidpour Aghjeh Mashhad, F., Mirdar Harijani, J., Gharaei, A. and Rahdari, A., 2016. Effect of different levels of tomato waste powder on blood serum biochemical parameters and growth indices of Common Carp Cyprinus carpio Linnaeus, 1758. Journal of Applied Ichthyological Research. 3(4): 101-114. (In Persian)
  30. Chitsaz, H. and Akrami, R., 2016. The effect of different levels of garlic skin on the activity of some liver enzymes of common carp (Cyprinus carpio), the first national conference of aromatic and spicy medicinal plants. Gonbad Kavos University. (In Persian)
  31. Maricchiolo, , Caruso, G. and Genovese, L., 2008. Haematological and Immunological Responses in Juvenile Sea Bass (Dicentrarchus labrax L.) After Short-Term Acute Stress, The Open Fish Science Journal. 1: 28-35
  32. Tort, L., Balasch J.C. and MacKenzie, S., 2004. Fish health challenge after stress. Indicators of immunocompetence. Contrib Sci. 2: 443-454.
  33. Mock, A. and Peters, G., 1990. Lysozyme activity in rainbow trout, Oncorhynchus mykiss (Walbaum), stressed by handling, transport and water pollution. J Fish Biol. 37: 873-885.
  34. Sabouri, S., Falahatkar, B., Khoshkholgh, M.R., Poursaeid, S. and Abtahi, B., 2017. Cortisol and Lactate dehydrogenase alternation in Caspian Kutum (Rutilus frisii) fingerlings exposed to crude oil pollution. Journal of Animal Research. 30(1): 79-89. (In Persian)
  35. Nezhadmoghadam, S., Imanpoor, M.R., Jafari, V. and Safari, R., 2018. Effect of Achillea millefolium on mucosal immune response and immune related (TNF-alfa) gene expression in Goldfish Carassius auratus (Linnaeus, 1758). Journal of Applied Ichthyological Research. 5(4): 87-100. (In Persian)
  36. Paterson,D., Rimmer, M.A., Keike, G.M. and Semmens, G.I., 2003. Physiological responses of the Asian sea bass, Lates calcarifer, to water quality deterioration during simulated live transport: acidosis, red-cell swelling, and levels of ions and ammonia in the plasma. Aquaculture. 218: 717-728. 10.1016/S0044-8486(02)00564-1.
  37. Raida, M.K. and Buchmann, K., 2008. Development of adaptive immunity in rainbow trout, Oncorhynchus mykiss (Walbaum) surviving an infection with Yersinia ruckeri. Fish Shellfish Immunol. 25(5):533-541. doi: 10.1016/j.fsi.2008.07.008. PMID: 18706505.
  38. Ackerman, P.A., Iwama, G.K. and Thornton, J.C., 2000. Physiological and immunological effects of adjuvanted Aeromonas salmonicida vaccines on juvenile rainbow trout. Journal of Aquatic Animal Health. 12(2): 157-164.
  39. Juha, K., Riitta, R., Marja, P. and Heikki, K., 2004. Effect of immunization with two commercial vaccines on feed intake, growth, and lysozyme activity in European whitefish (Coregonus lavaretus). Aquaculture. 234(1-4): 41-50.
  40. Huang, H.Y., Chena, Y.C., Wanga, P.C., Tsaia, M.A., Yeha, S.C., Liangb, H.J. and Chena, S.C., 2014. Efficacy of a formalin-inactivated vaccine against Streptococcus iniae infection in the farmed grouper Epinephelus coioides by intraperitoneal immunization. Vaccine. 32(51): 7014-7020.
  41. Safari, R., Moghadamfar, S., Imanpour, M.R., Shabani, A. and Jafar Nodeh, A., 2016. Antioxidant and immune gene expression in zebra fish (Danio rerio). Journal of Aquatic Ecology. 6(1): 93-101. (In Persian)
  42. Iwama, G. and Nakanishi, T., 1996. The Fish Immune System. Organism, Pathogen and Environment. Academic Press. 395 p.
  43. Salighehzadeh, R., Yavari, V., Mosavi, S. and Zakeri, M., 2016. Effect of Spirulina platensis as feed additive on immune factors in Mesopotamichthys sharpeyi. Journal of Aquatic Ecology. 5(1): 44-50. (In Persian)
  44. Grayfer, L., Walsh, J.G. and Belosevic, M., 2008. Characterization and functional analysis of goldfish tumornecrosis factor alfa. Developmental and Comparative Immunology. 32: 532-543.
  45. Jokinen, E.I.,Vielma, J., Aaltonen, T.M. and Koskela, J., 2003. The effect of dietary phosphorus deficiency on the immune responses of European whitefish (Coregonus lavaretus L.). Fish and Shellfish Immunology. 15(2): 159-168.
  46. Balfry, S.K., Maule, A.G. and Iwama, G.K., 2001. Coho salmon Oncorhynchus kisutch strain differences in disease resistance and non-specific immunity, following immersion challenges with Vibrio anguillarum, Diseases of Aquatic Organisms. 47(1): 39-48.
  47. Barnes, A.C., Young, F.M., Horne, M.T. and Ellis, A.E., 2003. Streptococcus iniae:  serological differences, presence of capsule and resistance to immune serum killing.  Inter-Research Diseases of Aquatic Organisms. 53(3): 241-247.
  48. Faghani, T., Azari Takami, Gh., Kousha, A. and Faghani, S., 2008. Surveying on Alginic Acid and Anti-Streptococcus Vaccine Effects on the Growth Performance, Survival Rate, Hematological Parameters in Rainbow Trout (Oncorhynchus mykiss). J. Zool. 3(2): 54-58.
  49. Badzohreh, Gh.R., Soltani, M., Shah Hoseini, Gh.R. and Nafisi Bahabadi, M., 2012. Effects of β-glucan on the growth, survival, and the efficacy of Anti-streptococcus iniae vaccine in rainbow trout (Onchorhynchus mykiss). Journal of Veterinary Research.67(1): 11-17. (In Persian)
  50. Rastiannasab, A., Mousavi, S., Zolgharnein, H. and Hosseinzadeh Sahafi, H., 2017. Effect of the food supplemented probioenzyme on expression of immune-related genes and redmouth disease (yersiniosis) control in rainbow trout (Oncorhynchus mykiss). Iranian Scientific Fisheries Journal. 26 (1) :153-166. (In Persian)
  51. Sheikhzadeh, N., Nootash, Sh., Khani Oushani, A., Mousavi, Sh. and Tahapour, K., 2019. Expression analysis of IFN-γ, MX1, MX2, MX3 and HSP70 genes in rainbow trout (Oncorhynchus mykiss) administrated with green tea (Camellia sinensis). Iranian Veterinary Journal. 15(3): 32-38. (In Persian)
  52. Hosseinifar, S.H., Hosseini, M., Paknezhad, H., Safari, R. and Jafer, A., 2018. The effect of retene and Lactobacillus acidophilus on nonspecific immune related gene expression in carp (cyprinus carpio). Journal of Utilization and Cultivation of Aquatics. 7(1): 35-41. (In Persian)
  53. Feng, M., Wang, Z.S., Zhou, A.G. and Ai, D.W., 2009. The effects of different sizes of nanometer zinc oxide on the proliferation and cell integrity of mice duodenum-epithelial cells in primary culture. Pakistan Journal of Nutrition. 8: 1164-1166.
  54. Iliev, D.B., Castellana, B., MacKenzie, S., Planas, J.V. and Goetz, F.W., 2007. Cloning and expression analysis of an IL-6 homolog in rainbow trout (Oncorhynchus mykiss). Molecular immunology 44: 1803-1807.
  55. Starkie, R., Ostrowski, S.R., Jauffred, S., Febbraio, M. and Pedersen, B.K., 2003. Exercise and IL‐6 infusion inhibit endotoxin‐induced TNF‐α production in humans. The Federation of American Societies for Experimental Biology. 17: 1-10.
  56. Jiang, , Chen, T., Sun, H., Tang, Z., Yu, J., Lin, Z., Ren, P.,  Zhou, X.,  Huang, Y., Li, X. and Yu, X., 2017. Immune response induced by oral delivery of Bacillus subtilis spores expressing enolase of Clonorchis sinensis in grass carps (Ctenopharyngodon idellus). Fish Shellfish Immunol. 60: 318-325. doi: 10.1016/j.fsi.2016.10.011. 
  57. Reyes-Becerril, M., Salinas, I., Cuesta, A., Meseguer, J., Tovar-Ramirez, D., Ascencio-Valle, F. and Ángeles Esteban, M., 2008. Oral delivery of live yeast Debaryomyces hansenii modulates the main innate immune parameters and the expression of immune-relevant genes in the gilthead seabream (Sparus aurata), Fish & Shellfish Immunology. 25(6): 731-739.
  58. Pérez-Sánchez, T., Balcázar, J.L., Merrifield, D.L., Carnevali, O., Gioacchini, G., de Blas, I. and Ruiz Zarzuela, I., 2011. Expression of immune-related genes in rainbow trout (Oncorhynchus mykiss) induced by probiotic bacteria during Lactococcus garvieae Fish & Shellfish Immunology. 31: 196-201.