Immunization of cultured shrimp against WSSV by use of produced recombinant vaccine(In Vitro)
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Seyed Mortezaei, S.R.
Mohseni Nejad, Lefteh
Seyfi Abadshapouri, Masoudreza
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AbstractShrimp diseases, especially viral diseases are the major limiting factors of shrimp farming activities in the world. White spot disease is one of the most pernicious viral disease that lead to serious loss in all shrimps of penaeids family. In Iran during 2002 to 2004 in Khuzestan province and in 2005 in Bushehr province, the most ponds and farms infected with white spot and the entire industry faced threat of closure. Surface proteins of virus has important role in the early stages of virus with host cell interaction, because these proteins usually enumerated as a candidate to produce a subunit and recombinant vaccines. On the other hand, these proteins can use as an important virus antigen and produce virus-specific antibodies in the preparation of diagnostic kits. In several studies proteins in the form of injection or oral vaccine has shown promising results. This study was designed in order to gain these proteins for immunization of white leg shrimp. To achieve the white spot virus genome, sampling and DNA extraction of moribund shrimps with clinical signs was done from Choebdeh farms. VP28 and VP19 protein-coding gene-specific primers were selected, and after amplification and purification were cloned in E.coli TG1. Protein expression evaluated and commercial plates were coated with inactivated bacteria containing the recombinant protein. White leg shrimp post larvae (PL30) were fed with recombinant food for 7 days then two times in 9th and 23th day after beginning of plate feeding and was challenged by white spot disease virus. The results of the first exposure (end of the seventh day after the shrimp fed with recombinant plate) showed that the lowest mortality at day 14 after exposure in different groups belong to VP28 group with cumulative mortality percent %30 ± 3.84 and the highest percentage of the Group TG1 equal to %72.22±2.93, also The highest and lowest relative percent survival, belong to the group of VP28 (60.29± 5.09%) and TG1 (4.41 ± 2.94%), respectively. In terms of mortality percent and relative percent survival, VP28 and VP 28+19 has significant difference with other groups while there was no significant difference between the groups VP19, TG1 and pMal. In the second exposure experiment (21 days after end of plate feeding) results showed that the lowest cumulative mortality in different groups belong to VP28 (%50 ± 5.09) and the highest percentage was owned by Group TG1 equal to %75.55±2.22, also the highest and the lowest of relative percent survival belong to VP28 (%32.86± 6.83) and TG1 (%0.00 ± 2.98) groups respectively. Similar to the first experiment, VP28 showed significant differences in cumulative mortality and relative percentage of survival in contrast to other group. As a general conclusion it can be noted that recombinant protein VP28 in any form that delivered to shrimp was able to protect shrimp against white spot virus. On the contrary VP19 has no this ability. The survival rate is directly related to the duration of the presence of recombinant protein in the diet.
Publisher or UniversityIranian Fisheries Science Research Institute
Series : Nr46114;