استفاده از روش های جدید برای به ثمر رساندن اثرات لاکتوباسیل ها به عنوان نسل جدید واکسن ها در صنعت آبزی پروری

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه بهداشت و بیماری های آبزیان، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران

2 گروه علوم دامی، پردیس کشاورزی و منابع طبیعی، دانشگاه تهران، کرج، ایران

3 گروه ژنتیک و مهندسی ژنتیک، موسسه تحقیقات واکسن و سرم سازی رازی، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرج، ایران

4 گروه بهداشت مواد غذایی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران

10.22034/aej.2021.280194.2496

چکیده

بیماری نکروز عفونی پانکراس (IPN) یک بیماری ویروسی قابل توجه در صنعت آبزی‌پروری است که گاها با وجود عدم مشاهده علایم بالینی منجر به ایجاد تلفات گسترده‌ای در ماهی می‌گردد، فلذا استفاده از روش‌های پیشگیری از جمله واکسیناسیون در این بیماری اهمیت بسیاری میابد. در این مطالعه قالب و الگوی DNA مورد استفاده برای تکثیر ژن VP2 محصول PCR، یک ایزوله ایرانی IPNV (IR-IPNV) بود و از Lactococcus lactis NZ9000 به عنوان میزبان کلونینگ و بیان ژن استفاده گردید. نتایج این مطالعه نشان دهنده افزایش معنی دار سطح آنتی‌بادی‌ها و علایم علایم بیماری در نتیجه مصرف لاکتوباسیلوس لاکتیس حاوی ژن در ماهی‌های بیمار بود (P<0/05). در این مطالعه برای نخستین بار جهت تولید واکسن نسل جدید نوترکیب از باکتری لاکتوباسیلوس لاکتیس برای کلون کردن ژن کاندید از پاتوژن عفونی در ماهی استفاده شد، بر اساس نتایج به دست آمده به نظر می‌رسد می‌توان از این سیستم بیانی به عنوان یک پلتفرم جدید جهت توسعه واکسن‌های خوراکی در ماهی‌ استفاده نمود که در صنعت آبزی پروری بسیار حائز اهمیت می باشد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Use of Lactococcus- expression system as oral recombinant vaccines as a new strategy

نویسندگان [English]

  • Asma Mohamadi 1
  • Mohammad Afsharnasab 1
  • Hamid Kohram 2
  • Khosrow Aghayipour 3
  • Abbass Ali Mottalebi 4
1 Department of Health, Aquatic Animal Health and Disease, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 Department of animal science, University college of agriculture and national resources, University of Tehran, Karaj, Iran.
3 Department of Genomics and Genetic Engineering, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran
4 Department of Food Hygiene, Science and Research Unit, Islamic Azad University, Tehran, Iran
چکیده [English]

Infectious pancreatic necrosis (IPN) is a significant viral disease in the aquaculture industry that sometimes leads to widespread mortality in fish despite the lack of clinical signs, so the use of prevention methods including Vaccination is very important in this disease. In this study, the DNA template and template used to amplify the VP2 gene of PCR product from an Iranian IPNV isolate (IR-IPNV). NZ9000 L. lactis strain was also used as host for cloning and as host for expression. The results showed that cloning and gene expression were well done and after the use of lactobacillus containing genes in the patient's fishes, it increased the antibody level and the viral infection decreased significantly. In this study, for the first time, L. lactis has been used to clone a candidate gene from an infectious pathogen of fish for the production of recombinant live vaccine vaccine. It seems that this expression system can be used to deliver an oral vaccine, Potential is considered as a new platform for the development of oral vaccines in fish.

کلیدواژه‌ها [English]

  • Recombinant live vector vaccine
  • Lactococcus lactis
  • Fish

مراجع

  1. Byrne, N., Castric, J., Lamour, F., Cabon, J. and Quentel, C., 2008. Study of the viral interference between infectious pancreatic necrosis virus (IPNV) and infectious haematopoietic necrosis virus (IHNV) in rainbow trout (Oncorhynchus mykiss). Fish & Shellfish Immunology. 24(5): 489-497.
  2. Bermúdez-Humarán, L.G., Aubry, C., Motta, J.P., Deraison, C., Steidler, L., Vergnolle, N., Chatel, J.M. and Langella, P., 2013. Engineering lactococci and lactobacilli for human health. Current Opinion in 16(3): 278-283.
  3. Biering, E., Villoing, S., Sommerset, I. and Christie, K., 2005. Update on viral vaccines for fish. Developments in Biologicals. 121: 97-113.
  4. Block, H., Maertens, B., Spriestersbach, A., Brinker, N., Kubicek, J., Fabis, R., Labahn, J. and Schäfer, F., 2009. Immobilized-metal affinity chromatography (IMAC): a review. Methods in Enzymology. 463: 439-473.
  5. Bowden, T., Smail, D. and Ellis, A., 2002. Development of a reproducible infectious pancreatic necrosis virus challenge model for Atlantic salmon, Salmo salar Journal of Fish Diseases. 25(9): 555-563.
  6. Cano-Garrido, O., Seras-Franzoso, J. and Garcia-Fruitós, E., 2015. Lactic acid bacteria: reviewing the potential of a promising delivery live vector for biomedical purposes. Microbial Cell Factories. 14(1): 1-12.
  7. Claassen, E., Winsen, R.V., Posno, M. and Boersma, W., 1995. New and safe" oral" live vacines based on lactobacillus. Advances in Experimental Medicine and Biology. 371: 1553.
  8. Crane, M.S.J., Hardy-Smith, P., Williams, L.M., Hyatt, A.D., Eaton, L.M., Gould, A., Handlinger, J., Kattenbelt, J. and Gudkovs, N., 2000. First isolation of an aquatic birnavirus from farmed and wild fish species in Australia. Diseases of Aquatic Organisms. 43(1): 1-14.
  9. Cuesta, A., Chaves-Pozo, E., de Las Heras, A., Saint-Jean, S.R., Perez-Prieto, S. and Tafalla, C., 2010. An active DNA vaccine against infectious pancreatic necrosis virus (IPNV) with a different mode of action than fish rhabdovirus DNA vaccines. Vaccine. 28(19): 3291-3300.
  10. Dadar, M., Rajabi Memari, H., Peyghan, R. and Seifi Abad Shapouri, M., 2014. Isolation and expression of recombinant viral protein (VP2) from Iranian isolates of Infectious Pancreatic Necrosis Virus (IPNV) in Escherichia coli. Iranian Journal of Fisheries Sciences. 13(4): 856-868.
  11. Davies, K.R., McColl, K.A., Wang, L.F., Yu, M., Williams, L.M. and Crane, M.S.J., 2010. Molecular characterisation of Australasian isolates of aquatic birnaviruses. Diseases of Aquatic Organisms. 93(1): 1-15.
  12. De Ruyter, P., Kuipers, O.P., Beerthuyzen, M.M., van Alen-Boerrigter, I. and de Vos, W.M., 1996. Functional analysis of promoters in the nisin gene cluster of Lactococcus lactis. Journal of Bacteriology. 178(12): 3434-3439.
  13. Dhar, A.K., Bowers, R.M., Rowe, C.G. and Allnutt, F.T., 2010. Expression of a foreign epitope on infectious pancreatic necrosis virus VP2 capsid protein subviral particle (SVP) and immunogenicity in rainbow trout. Antiviral Research. 85(3): 525-531.
  14. Dobos, P., 1995. The molecular biology of infectious pancreatic necrosis virus (IPNV). Annual Review of Fish Diseases. 5: 25-54.
  15. Dobos, P., Hill, B., Hallett, R., Kells, D., Becht, H. and Teninges, D., 1979. Biophysical and biochemical characterization of five animal viruses with bisegmented double-stranded RNA genomes'. Journal of Virology. 32(2): 593-605.
  16. Dolatbadi, S., Falsafi, T. and Mahmoudi, M., 2015. Lactococcus lactis as an oral vector for cloning of heat shock protein a from Helicobacter pylori. International Journal of Biosciences. 6: 410-415.
  17. Duncan, R., Mason, C.L., Nagy, E., Leong, J.A. and Dobos, P., 1991. Sequence analysis of infectious pancreatic necrosis virus genome segment B and its encoded VP1 protein: a putative RNA-dependent RNA polymerase lacking the Gly-Asp-Asp motif. Virology. 181(2): 541-552.
  18. Embregts, C.W. and Forlenza, M., 2016. Oral vaccination of fish: Lessons from humans and veterinary species. Developmental & Comparative Immunology. 64: 118-137.
  19. Evensen, Ø. and Leong, J.A., 2013. DNA vaccines against viral diseases of farmed fish., Fish & Shellfish Immunology. Vol. 35, No. 6, pp. 1751-1758.
  20. Fridholm, H., Eliasson, L. and Everitt, E., 2007. Immunogenicity properties of authentic and heterologously synthesized structural protein VP2 of infectious pancreatic necrosis virus. Viral Immunology. 20(4): 635-648.
  21. Frost, P., Börsheim, K. and Endresen, C., 1998. Analysis of the antibody response in Atlantic salmon against recombinant VP2 of infectious pancreatic necrosis virus (IPNV). Fish & Shellfish Immunology. 8(6): 447-456.
  22. Frost, P. and Ness, A., 1997. Vaccination of Atlantic salmon with recombinant VP2 of infectious pancreatic necrosis virus (IPNV), added to a multivalent vaccine, suppresses viral replication following IPNV challenge. Fish & Shellfish Immunology. 7(8): 609-619.
  23. Ahmadivand, S., Soltani, M., Behdani, M., Evensen, Ø., Alirahimi, E., Hassanzadeh, R. and Soltani, E., 2017. Oral DNA vaccines based on CS-TPP nanoparticles and alginate microparticles confer high protection against infectious pancreatic necrosis virus (IPNV) infection in trout. Developmental & Comparative Immunology. 74: 178-189.
  24. Ahmadi, N., Oryan, A., Akhlaghi, M. and Hosseini, A., 2013. Tissue distribution of infectious pancreatic necrosis virus serotype S p in naturally infected cultured rainbow trout, Oncorhynchus mykiss (Walbaum): an immunohistochemical and nested‐PCR study. Journal of Fish Diseases. 36(7): 629-637.
  25. Akhlaghi, M. and Hosseini, A., 2007. First report on the detection of infectious pancreatic necrosis virus (IPNV) by RT-PCR in rainbow trout fry cultured in Iran. Bulletin-European Association of Fish Pathologists. 27(5): 205.
  26. Allnutt, F.T., Bowers, R.M., Rowe, C.G., Vakharia, V.N., LaPatra, S.E. and Dhar, A.K., 2007. Antigenicity of infectious pancreatic necrosis virus VP2 subviral particles expressed in yeast. Vaccine. 25(26): 4880-4888.
  27. Ana, I., Prieto, S.I.P. and Saint-Jean, S.R., 2009. In vitro and in vivo immune responses induced by a DNA vaccine encoding the VP2 gene of the infectious pancreatic necrosis virus. Fish & Shellfish Immunology. 27(2): 120-129.
  28. Heras, A., Saint-Jean, S. and Pérez-Prieto, S., 2010. Immunogenic and protective effects of an oral DNA vaccine against infectious pancreatic necrosis virus in fish. Fish Shellfish Immunol. 28(4): 562-570.
  29. Ballesteros, N.A., Saint-Jean, S.R. and Perez-Prieto, S.I., 2015. Immune responses to oral pcDNA-VP2 vaccine in relation to infectious pancreatic necrosis virus carrier state in rainbow trout Oncorhynchus mykiss. Veterinary Immunology and Immunopathology. 165(3-4): 127-137.
  30. Benson, L.M., Null, A.P. and Muddiman, D.C., 2003. Advantages of Thermococcus kodakaraenis (KOD) DNA polymerase for PCR-mass spectrometry based analyses. Journal of the American Society for Mass Spectrometry. 14(6): 601-604.
  31. Galdeano, C.M. and Perdigon, G., 2006. The probiotic bacterium Lactobacillus casei induces activation of the gut mucosal immune system through innate immunity. Clinical and Vaccine Immunology. 13(2): 219-226.
  32. Gareau, M.G., Sherman, P.M. and Walker, W.A., 2010. Probiotics and the gut microbiota in intestinal health and disease. Nature Reviews Gastroenterology & Hepatology. 7(9): 503.
  33. Gudding, R., Lillehaug, A. and Evensen, Ø., 2014. Fish vaccination, Wiley Online Library, New York.
  34. Hall, T.A. 2000. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT'. Information Retrieval Ltd., c1979-c2000. London. 95-98.
  35. Hølvold, L.B., Myhr, A.I. and Dalmo, R.A., 2014. Strategies and hurdles using DNA vaccines to fish. Veterinary Research. 45(1): 1-11.
  36. Hou, X.L., Yu, L.Y., Liu, J. and Wang, G.H., 2007. Surface-displayed porcine epidemic diarrhea viral (PEDV) antigens on lactic acid bacteria. Vaccine. 26(1): 24-31.
  37. Kleerebezem, M., Beerthuyzen, M.M., Vaughan, E.E., De Vos, W.M. and Kuipers, O.P., 1997. Controlled gene expression systems for lactic acid bacteria: transferable nisin-inducible expression cassettes for Lactococcus, Leuconostoc, and Lactobacillus Applied and environmental Microbiology. 63(11): 4581-4584.
  38. Konings, W.N., Kok, J., Kuipers, O.P. and Poolman, B., 2000. Lactic acid bacteria: the bugs of the new millennium. Current Opinion in Microbiology. 3(3): 276-282.
  39. Kurath, G., 2008. Biotechnology and DNA vaccines for aquatic animals. Revue Scientifique et Technique-Office International des Épizooties. 27(1): 175.
  40. Labus, M.B., Breeman, S., Ellis, A.E., Smail, D.A., Kervick, M. and Melvin, W.T., 2001. Antigenic comparison of a truncated form of VP2 of infectious pancreatic necrosis (IPN) virus expressed in four different cell types. Fish & Shellfish Immunology. 11(3): 203-216.
  41. Lam, D.M.K. and Xu, Y., 2013. Oral vaccines produced and administered using edible micro-organisms including lactic acid bacterial strains. United States Patent Application US, New York.
  42. LeBlanc, J.G., Aubry, C., Cortes-Perez, N.G., de Moreno de LeBlanc, A., Vergnolle, N., Langella, P., Azevedo, V., Chatel, J.M., Miyoshi, A. and Bermúdez-Humarán, L.G., 2013. Mucosal targeting of therapeutic molecules using genetically modified lactic acid bacteria: an update. FEMS Microbiology Letters. 344(1): 1-9.
  43. Li-Li, Z., Min, L., Jun-Wei, G., Xin-Yuan, Q., Yi-Jing, L. and Di-Qiu, L., 2012. Expression of infectious pancreatic necrosis virus (IPNV) VP2–VP3 fusion protein in Lactobacillus casei and immunogenicity in rainbow trouts. Vaccine. 30(10): 1823-1829.
  44. Liu, C.Q., Su, P., Khunajakr, , Deng, Y.M., Sumual, S., Kim, W., Tandianus, J. and Dunn, N., 2005. Development of food‐grade cloning and expression vectors for Lactococcus lactis. Journal of Applied Microbiology. 98(1): 127-135.
  45. Liu, D., Wang, X., Ge, J., Liu, S. and Li, Y., 2011. Comparison of the immune responses induced by oral immunization of mice with Lactobacillus casei-expressing porcine parvovirus VP2 and VP2 fused to Escherichia coli heat-labile enterotoxin B subunit protein. Comparative Immunology, Microbiology and Infectious Diseases. 34(1): 73-81.
  46. Lorenzen, N. and LaPatra, S., 2005. DNA vaccines for aquacultured fish. International Office of Epizootics. 24(1): 201-213.
  47. Manríquez, R.A., Vera, T., Villalba, M.V., Mancilla, A., Vakharia, V.N., Yañez, A.J. and Cárcamo, J.G., 2017. Molecular characterization of infectious pancreatic necrosis virus strains isolated from the three types of salmonids farmed in Chile. Virology Journal. 14(1): 1-16.
  48. Mierau, I. and Kleerebezem, M., 2005. 10 years of the nisin-controlled gene expression system (NICE) in Lactococcus lactis'. Applied Microbiology and Biotechnology. 68(6): 705-717.
  49. Mierau, I., Leij, P., Van Swam, I., Blommestein, B., Floris, E., Mond, J. and Smid, E.J., 2005. Industrial-scale production and purification of a heterologous protein in Lactococcus lactis using the nisin-controlled gene expression system NICE: the case of lysostaphin. Microbial Cell Factories. 4(1): 1-9.
  50. Min, L., Li-Li, Z., Jun-Wei, G., Xin-Yuan, Q., Yi-Jing, L. and Di-Qiu, , 2012. Immunogenicity of Lactobacillus-expressing VP2 and VP3 of the infectious pancreatic necrosis virus (IPNV) in rainbow trout. Fish & Shellfish Immunology. 32(1): 196-203.
  51. Molloy, S.D., Pietrak, M.R., Bricknell, I. and Bouchard, D.A., 2013. Experimental transmission of infectious pancreatic necrosis virus from the blue mussel, Mytilus edulis, to cohabitating Atlantic salmon (Salmo salar) smolts. Applied and Environmental Microbiology. 79(19): 5882-5890.
  52. Munang’andu, H.M. and Evensen, Ø., 2015. A review of intra-and extracellular antigen delivery systems for virus vaccines of finfish. Journal of Immunology Research. 2015:960859. doi: 10.1155/2015/960859.
  53. Null, A.P. and Muddiman, D.C., 2001. Perspectives on the use of electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry for short tandem repeat genotyping in the post‐genome era. Journal of Mass Spectrometry. 36(6): 589-606.
  54. Offerman, J.D. and Rychlik, W., 2003. Oligo primer analysis software. Introduction to Bioinformatics, Springer, New York. 345-355.
  55. Ogawa, T., Asai, Y., Sakamoto, H. and Yasuda, K., 2006. Oral immunoadjuvant activity of Lactobacillus casei subsp. casei in dextran-fed layer chickens. British Journal of Nutrition. 95(2): 430-434.
  56. Oryan, A., Ahmadi, N., Akhlaghi, M. and Hosseini, A., 2012. A comparative histopathological, immunohistochemical and nested-PCR study for diagnosis of infectious pancreatic necrosis in the naturally infected cultured rainbow trout (Oncorhynchus mykiss). Aquaculture International. 20(4): 725-734.
  57. Petit, S., Lejal, N., Huet, J.C. and Delmas, B., 2000. Active residues and viral substrate cleavage sites of the protease of the birnavirus infectious pancreatic necrosis virus. Journal of Virology. 74(5): 2057-2066.
  58. Platteeuw, C., van Alen-Boerrigter, I., van Schalkwijk, S. and De Vos, W., 1996. Food-grade cloning and expression system for Lactococcus lactis. Applied and Environmental Microbiology. 62(3): 1008-1013.
  59. Pouwels, P.H., Leer, R.J. and Boersma, W.J., 1996. The potential of Lactobacillus as a carrier for oral immunization: development and preliminary characterization of vector systems for targeted delivery of antigens. Journal of Biotechnology. 44(1-3): 183-192.
  60. Raissy, M., Momtaz, H., Ansari, M., Moumeni, M. and Hosseinifard, M., 2010. Distribution of infectious pancreatic necrosis virus (IPNV) in two major rainbow trout fry producing provinces of Iran with respect to clinically infected farms. Journal of Food, Agriculture & Environment. 8(3/4): 614-615.
  61. Ramstad, A. and Midtlyng, P., 2008. Strong genetic influence on IPN vaccination‐and‐challenge trials in Atlantic salmon, Salmo salar Journal of Fish Diseases. 31(8): 567-578.
  62. Ramstad, A., Romstad, A., Knappskog, D. and Midtlyng, P., 2007. Field validation of experimental challenge models for IPN vaccines. Journal of Fish Diseases. 30(12): 723-731.
  63. Roberts, R. and Pearson, M., 2005. Infectious pancreatic necrosis in Atlantic salmon, Salmo salar Journal of Fish Diseases. 28(7): 383-390.
  64. Rønneseth, A., Wergeland, H.I., Devik, M., Evensen, Ø. and Pettersen, E.F., 2007. Mortality after IPNV challenge of Atlantic salmon (Salmo salar) differs based on developmental stage of fish or challenge route. Aquaculture. 271(1-4): 100-111.
  65. Ross, R.P., Morgan, S. and Hill, C., 2002. Preservation and fermentation: past, present and future. International Journal of Food Microbiology. 79(1-2): 3-16.
  66. Shivappa, R., McAllister, P., Edwards, G. and Santi, N., 2005. Using a baculovirus insect/larvae. Developments in Biologicals. 121: 165-174.
  67. Smail, D., Bain, N., Bruno, D., King, J., Thompson, F., Pendrey, D., Morrice, S. and Cunningham, C., 2006. Infectious pancreatic necrosis virus in Atlantic salmon, Salmo salar, post‐smolts in the Shetland Isles, Scotland: virus identification, histopathology, immunohistochemistry and genetic comparison with Scottish mainland isolates. Journal of Fish Diseases. 29(1): 31-41.
  68. Smail, D., Bruno, D., Dear, G., McFarlene, L. and Ross, K., 1992. Infectious pancreatic necrosis (IPN) virus Sp serotype in farmed Atlantic salmon, Salmo salar, post-smolts associated with mortality and clinical disease. Journal of Fish Diseases. 15(1): 77-83.
  69. Smith, H.A., 1994. Regulatory considerations for nucleic acid vaccines. Vaccine. 12(16): 1515-1519.
  70. Stagg, R. and Skjelstad, B., 2003. Control of IPN', IPN in salmonids: a review. Fisheries and Aquaculture Industries Research Fund Oslo. 4: 10-16.
  71. Takagi, M., Nishioka, M., Kakihara, H., Kitabayashi, M., Inoue, H., Kawakami, B., Oka, M. and Imanaka, T., 1997. Characterization of DNA polymerase from Pyrococcus strain KOD1 and its application to PCR. Applied and Environmental Microbiology. 63(11): 4504-4510.
  72. Tapia, D., Eissler, Y., Torres, P., Jorquera, E., Espinoza, J. and Kuznar, J., 2015. Detection and phylogenetic analysis of infectious pancreatic necrosis virus in Chile. Diseases of Aquatic Organisms. 116(3): 173-184.
  73. Tarrab, E., Berthiaume, L., Grothe, S., O'Connor-McCourt, M., Heppell, J. and Lecomte, J., 1995. Evidence of a major neutralizable conformational epitope region on VP2 of infectious pancreatic necrosis virus. Journal of General Virology. 76(3): 551-558.
  74. Temin, H., 1998. Overview of biological effects of addition of DNA molecules to cells. Developments in Biological Standardization. 93: 37-44.
  75. Tonheim, T.C., Bøgwald, J. and Dalmo, R.A., 2008. What happens to the DNA vaccine in fish? A review of current knowledge. Fish & Shellfish Immunology. 25(1-2): 1-18.
  76. Villatoro-Hernández, J., Kuipers, O.P., Saucedo-Cárdenas, O. and Montes-de-Oca-Luna, R., 2012. 'Heterologous protein expression by Lactococcus lactis', in Recombinant gene expression, Springer, New York. 155-165.
  77. Villena, J., Salva, S., Agüero, G. and Alvarez, S., 2011. Immunomodulatory and protective effect of probiotic Lactobacillus casei against Candida albicans infection in malnourished mice. Microbiology and Immunology. 55(6): 434-445.
  78. Wells, J., 2011. Mucosal vaccination and therapy with genetically modified lactic acid bacteria. Annual Review of Food Science and Technology. 2: 423-445.
  79. Wolf, K., Snieszko, S., Dunbar, C. and Pyle, E., 1960. Virus nature of infectious pancreatic necrosis in trout. Proceedings of the Society for Experimental Biology and Medicine. 104(1): 105-108.
  80. Wolff, J.A., Ludtke, J.J., Acsadi, G., Williams, P. and Jani, A., 1992. Long-term persistence of plasmid DNA and foreign gone expression in mouse muscle. Human Molecular Genetics. 1(6): 363-369.
  81. Zhu, L., Wang, X., Wang, K., Yang, Q., He, J., Qin, Z., Geng, Y., Ouyang, P. and Huang, X., 2017. Outbreak of infectious pancreatic necrosis virus (IPNV) in farmed rainbow trout in China. Acta Tropica. 170: 63-69.
فصلنامه محیط زیست جانوری
دوره 14، شماره 1
شماره بهار
اردیبهشت 1401
صفحه 441-450

فایل ها

هم رسانی

ارجاع به این مقاله

آمار