Methods and compositions to protect aquatic invertebrates from disease
Compositions and methods of protecting aquatic invertebrates from disease is shown. In one embodiment, dsRNA or antisense RNA to a nucleic acid molecule of the disease-causing microorganism is prepared and delivered to the animal. In another embodiment, a nucleic acid molecule of the disease-causing microorganism is delivered to the animal. In another embodiment, the RNA or nucleic acid molecule is delivered to the animal by replicon particle. In a further embodiment, the protective molecule is delivered to the digestive tract of the animal. Protection from disease is obtained.
1. A method of producing a composition for protecting shrimp from disease, comprising,
a) identifying a target nucleic acid molecule of a microorganism causing said disease;
b) preparing a protective molecule selected from the group consisting of:
(i) a dsRNA molecule comprising at least one RNA molecule that is capable of hybridizing to said target nucleic acid molecule of said microorganism;
(ii) an antisense RNA molecule comprising a sequence that is the full or partial complement of said target nucleic acid molecule of said microorganism; and
c) preparing an orally acceptable composition comprising said protective molecule with feed, wherein said composition does not comprise an expression cassette or vector comprising said protective molecule, and wherein when orally administered to said shrimp said composition protects said shrimp from said disease.
2. The method of claim 1 , wherein said at least one RNA molecule comprising a sequence that corresponds to all or a portion of said target nucleic acid molecule is selected from the group consisting of an RNA molecule of at least 10 bp, at least 20 bp, at least 30 bp and at least 50 bp.
3. The method of claim 1 , wherein said animal is protected from said disease for a period of time selected from the group consisting of at least 20 days, at least 25 days, at least 30 days, at least 40 days and at least 60 days following administration of said protective molecule.
4. The method of claim 1 , wherein said dsRNA comprises at least one RNA molecule that is the full or partial complement of all or a portion of said target nucleic acid molecule.
5. The method of claim 1 , wherein said disease comprises White Spot Syndrome Virus (WSSV) or Infectious Myonecrosis Virus (IMNV).
6. The method of claim 1 , wherein said target nucleic acid molecule comprises a nucleic acid molecule encoding White Spot Syndrome Virus (WSSV) VP19 or VP28 or a fragment thereof, or Infectious Myonecrosis Virus (IMNV) or a fragment thereof, wherein said nucleic acid molecule or fragment thereof produces a protective molecule that when administered to said shrimp, protects said shrimp from said WSSV or IMNV.
7. The method of claim 1 , wherein said target molecule comprises a nucleic acid molecule encoding White Spot Syndrome Virus (WSSV) VP19 or VP28.
8. The method of claim 1 , wherein said target nucleic acid molecule comprises SEQ ID NO: 29 or a fragment thereof or SEQ ID NO: 30 or a fragment thereof.
9. The method of claim 1 , wherein said sequence that corresponds to all or a portion of said target nucleic acid comprises SEQ ID NO: 47 or a fragment thereof, SEQ ID NO: 49 or a fragment thereof, and said RNA that is the full or partial complement of all or a portion of said target nucleic acid molecule comprises SEQ ID NO: 48 or a fragment thereof or SEQ ID NO: 50 or a fragment thereof.
10. The method of claim 1 , wherein said dsRNA is produced by a nucleic acid molecule comprising SEQ ID NO: 32, or the complement thereof, or a fragment of said nucleic acid molecule, wherein said dsRNA produces a protective response.
11. The method of claim 1 , wherein said disease comprises Infectious Myonecrosis Virus (IMNV).
12. The method of claim 1 , wherein said sequence that corresponds to all or a portion of said target nucleic acid molecule is selected from the group consisting of SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 52, SEQ ID NO: 57, and SEQ ID NO: 62 and a fragment thereof, and said RNA that is the full or partial complement of all or a portion of said target nucleic acid molecule is selected from the group consisting of SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 53, SEQ ID NO: 58, and SEQ ID NO: 63 and a fragment thereof.
13. The method of claim 1 , wherein said dsRNA is produced by a nucleic acid molecule selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 32, SEQ ID NO: 51, SEQ ID NO: 56, SEQ ID NO: 61 and SEQ ID NO: 76 or the complement thereof, and a fragment of said nucleic acid molecule wherein said dsRNA produces a protective response.
14. The method of claim 1 , wherein said dsRNA comprises a sequence that corresponds to all or a portion of said target nucleic acid molecule and a sequence that is the full or partial complement of said target nucleic acid molecule, wherein said sequence that corresponds to all or a portion of said target nucleic acid molecule is selected from the group consisting of SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 52, SEQ ID NO: 57, and SEQ ID NO: 62 and a fragment thereof, and said RNA that is the full or partial complement of all or a portion of said target nucleic acid molecule is selected from the group consisting of SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 53, SEQ ID NO: 58, and SEQ ID NO: 63 and a fragment thereof.
15. The method of claim 1 , wherein said target molecule comprises a nucleotide sequence encoding SEQ ID NO: 68 or fragment thereof wherein said protective molecule when administered to said animal, protects said animal from IMNV.
16. The method of claim 1 , wherein said target molecule comprises SEQ ID NO: 72 or fragment thereof wherein said protective molecule when administered to said shrimp, protects said shrimp from IMNV.
17. A method of producing a composition for protecting shrimp from White Spot Syndrome Virus (WSSV) or Infectious Myonecrosis Virus (IMNV), comprising,
a) identifying a target nucleic acid molecule of said WSSV or IMNV;
b) preparing a protective molecule selected from the group consisting of:
(i) a dsRNA molecule comprising at least one RNA molecule that is capable of hybridizing to said target nucleic acid molecule of said WSSV or IMNV;
(ii) an antisense RNA molecule comprising a sequence that is the full or partial complement of said target nucleic acid molecule of said WSSV or IMNV; and
c) preparing an orally acceptable composition comprising said protective molecule with feed, wherein does not comprise an expression cassette or vector comprising said protective molecule, and wherein when orally administered to said shrimp said composition protects said shrimp from said WSSV or IMNV.
18. A method of producing a composition for protecting shrimp from disease, comprising,
a) identifying a target nucleic acid molecule of a microorganism causing said disease;
b) preparing a protective molecule selected from the group consisting of:
(i) a dsRNA molecule comprising at least one RNA molecule that is capable of hybridizing to said target nucleic acid molecule of said microorganism;
(ii) an antisense RNA molecule comprising a sequence that is the full or partial complement of said target nucleic acid molecule of said microorganism; and
c) preparing an orally acceptable composition for delivery to the digestive tract of said shrimp, said composition comprising said protective molecule with feed, wherein said composition does not comprise an expression cassette or vector comprising said protective molecule, and wherein when orally administered to said shrimp said composition protects said shrimp from said disease.
19. The method of claim 17 wherein said orally acceptable composition comprising said protective molecule with feed comprises feed coated with said protective molecule.
20. The method of claim 1 wherein said orally acceptable composition comprising said protective molecule with feed comprises feed coated with said protective molecule.