Circovirus sequences associated with piglet weight loss disease (pwd)

Abstract

The genome sequences and the nucleotide sequences coding for the PWD circovirus polypeptides, such as the circovirus structural and non-structural polypeptides, vectors including the sequences, and cells and animals transformed by the vectors are provided. Methods for detecting the nucleic acids or polypeptides, and kits for diagnosing infection by a PWD circovirus, also are provided. Method for selecting compounds capable of modulating the viral infection is further provided. Pharmaceutical, including vaccines, compositions for preventing and/or treating viral infections caused by PWD circovirus and the use of vectors for preventing and/or treating diseases also are provided.

1 . A vaccine for the prevention or treatment of a viral infection by a piglet weight loss disease circovirus. 
     
     
         2 . A vaccine for the prevention or treatment of a viral infection by a piglet weight loss disease circovirus, wherein said vaccine comprises a nucleotide sequence from the genome of porcine circovirus type B, or a homolog or fragment thereof 
     
     
         3 . The vaccine of  claim 2 , wherein said nucleotide sequence is selected from SEQ ID NO:15, SEQ ID NO:19, SEQ ID NO:23, and SEQ ID NO:25. 
     
     
         4 . The vaccine of  claim 2 , wherein said homolog has at least 80% sequence identity to SEQ ID NO:15, SEQ ID NO:19, SEQ ID NO:23, or SEQ ID NO:25. 
     
     
         5 . A vaccine comprising a polypeptide encoded by a nucleotide sequence from the genome of porcine circovirus type B. 
     
     
         6 . The vaccine of  claim 5 , further comprising a pharmaceutical or veterinary vehicle. 
     
     
         7 . The vaccine of  claim 5 , where said polypeptide has the amino acid sequence selected from SEQ ID NO:24 and SEQ ID NO:26. 
     
     
         8 . A method of immunizing an animal against piglet weight loss disease, comprising administering to an animal an effective amount of a vaccine, wherein said vaccine comprises a nucleotide sequence from the genome of porcine circovirus type B. 
     
     
         9 . A method of immunizing an animal against piglet weight loss disease, comprising administering to an animal an effective amount of a vaccine, wherein said vaccine comprises a polypeptide encoded by a nucleotide sequence from the genome of porcine circovirus type B. 
     
     
         10 . An immunogenic agent comprising a polypeptide selected from SEQ ED NO:29, SEQ ID NO:30, SEQ ID NO:31, and SEQ ID NO:32. 
     
     
         11 . A medicament comprising the immunogenic agent of  claim 10 . 
     
     
         12 . The vaccine of  claim 1 , comprising at least one antigen of a porcine circovirus type B. 
     
     
         13 . A multivalent combination vaccine, comprising an immunological agent effective for reducing the incidence of or lessening the severity of porcine circovirus type B (PVCB) infections in comparison to a swine not receiving said multivalent combination vaccine and at least one immunogenic active component effective against another disease-causing organism in swine, wherein said multivalent combination vaccine is effective at reducing the incidence of or lessening the severity of PCVB infections for at least 25 days after the administration of a single dose of said multivalent combination vaccine. 
     
     
         14 . The multivalent combination vaccine of  claim 13 , wherein said immunological agent effective for reducing the incidence of or lessening the severity of PCVB infection is a PCVB antigen. 
     
     
         15 . The multivalent combination vaccine of  claim 14 , wherein the PCVB antigen is inactivated PCVB antigen, a live modified or attenuated PCVB, a chimeric virus that comprises at least an immunogenic amino acid sequence of PCVB, or any other polypeptide or component that comprises at least an immunogenic amino acid sequence of PCVB. 
     
     
         16 . The multivalent combination vaccine of  claim 14 , wherein said PCVB antigen is a protein encoded by a DNA sequence having at least 80% sequence identity with ORF′2 of a PCVB. 
     
     
         17 . The multivalent combination vaccine of  claim 14 , wherein said PCVB antigen is a baculovirus expressed ORF′2 antigen. 
     
     
         18 . The multivalent combination vaccine of  claim 13 , wherein said other disease-causing organism is a virus, bacteria, parasite, or prion. 
     
     
         19 . The multivalent combination vaccine of  claim 13 , wherein said other disease-causing organism is a virus. 
     
     
         20 . The multivalent combination vaccine of  claim 13 , wherein said other disease-causing organism is porcine circovirus type A (PVCA). 
     
     
         21 . The multivalent combination vaccine of  claim 13 , wherein said other disease-causing organism is a pestivirus or arterivirus. 
     
     
         22 . The multivalent combination vaccine of  claim 13 , wherein said other disease-causing organism causes Aujesky's disease or porcine dysgenic and respiratory syndrome (PDRS) in swine. 
     
     
         23 . The multivalent combination vaccine of  claim 13 , wherein said effective component comprises an attenuated living microorganism, an inactivated microorganism, a cellular extract, a recombined protein, or a polysaccharide. 
     
     
         24 . The multivalent combination vaccine of  claim 13 , wherein said other disease-causing organism is  M. bovis, Bordatella pertussis  or  Pneumococcus.    
     
     
         25 . The multivalent combination vaccine of  claim 13 , wherein said other disease-causing organism is a hepatitis virus or an influenza virus. 
     
     
         26 . The multivalent combination vaccine of  claim 13 , wherein said immunogenic active component effective against another disease-causing organism in swine is selected from the group consisting of an antigen of:  Actinobacillus pleuropneumonia ; Adenovirus; Alphavirus such as Eastern equine encephalomyelitis viruses;  Bordetella bronchiseptica; Brachyspira  spp., preferably  B. hyodyentheria; B. piosicoli, Brucella suis , preferably biovars 1, 2, and 3: Clasical swine fever virus;  Clostridium  spp., preferably  Cl. difficile, Cl. perfringens  types A, B, and C,  Cl. novyi, Cl. septicum, Cl. tetani ; Coronavirus, preferably Porcine Respiratory Corona virus; Eperythrozoonosis suis; Erysipelothrix rhsiopathiae;  Escherichia coli; Haemophilus parasuis , preferably subtypes 1, 7 and 14; Hemagglutinating encephalomyelitis virus; Japanese Encephalitis Virus;  Lawsonia intracellularis; Leptospira  spp., preferably  Leptospira australis; Leptospira canicola; Leptospira grippotyphosa; Leptospira icterohaemorrhagicae ; and  Leptospira intenogans; Leptospira pomona; Leptospira tarassovi; Mycobacterium  spp., preferably  M. avium, M. intracellulare  and  M. bovis; Mycoplasma hyopneumoniae  (M hyo);  Pasteurella multocida; Porcine cytomegalovirus; Porcine Parvovirus ; Porcine Reproductive and Respiratory Syndrome (PRRS) Virus, Pseudorabies virus; Rotavirus;  Salmonella  spp., preferably  S. thyhimurium  and  S. choleraesuis; Staph. hyicus; Staphylococcus  spp., preferably  Streptococcus  spp., preferably  Strep. suis ; Swine herpes virus; Swine Influenza Virus; Swine pox virus; Swine pox virus. Vesicular stomatitis virus; Virus of vesicular exanthema of swine, and combinations thereof 
     
     
         27 . A multivalent immunogenic composition comprising:
 an antigen of PCVB; and   at least one antigen from another disease-causing organism in swine, wherein said antigen of PCVB reduces the incidence of or lessens the severity of PCVB infections in comparison to an animal not receiving said immunogenic composition for at least 25 days after the administration of a single dose of said multivalent immunogenic composition.   
     
     
         28 . A multivalent immunogenic composition comprising:
 an antigen of PCVB; and   at least one antigen from another disease-causing organism in swine, wherein said antigen of PCVB reduces the incidence of tissues positive for signs of PCVB infection in comparison to an animal not receiving said multivalent immunogenic composition as determined by immunohistochemical testing for at least 25 days after the administration of a single dose of said multivalent immunogenic composition.   
     
     
         29 . A multivalent immunogenic composition comprising:
 i) an antigenic protein of PCVB;   ii) at least one further antigen from another disease-causing organism in swine; and   iii) an adjuvant.   
     
     
         30 . The multivalent immunogenic composition according to  claim 29 , wherein said protein of PCVB reduces the incidence of tissues positive for the signs of PCVB infection in comparison to an animal not receiving said multivalent immunogenic composition as determined by immunohistochemical testing for at least 25 days after the administration of a single dose of said multivalent immunogenic composition. 
     
     
         31 . The multivalent immunogenic composition according to  claim 29 , wherein said antigenic protein of PCVB reduces the incidence of or lessens the severity of PCVB infections when administered to an animal, wherein said reduction of the incidence of or lessening the severity is in comparison to an animal not receiving said immunogenic composition for at least 25 days after the administration of a single dose of said multivalent immunogenic composition. 
     
     
         32 . The immunogenic composition according to  claim 29 , wherein said adjuvant is aluminum hydroxide, a muramyl peptide, a bacterial lysate, Freund's incomplete adjuvant, AIF™, or AIF SEPPIC. 
     
     
         33 . The immunogenic composition according to  claim 29 , wherein said antigenic protein is selected from the group consisting of:
 i) a polypeptide comprising the sequence of SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:55, SEQ ID NO:58, or SEQ ID NO:123;   ii) a polypeptide that is at least 90% homologous to the polypeptide of i);   iii) a polypeptide that is encoded by a DNA comprising the sequence of SEQ ID NO:15, SEQ ID NO:19, or SEQ ID NO:25; and   iv) a polypeptide that is encoded by a polynucleotide that is at least 90% homologous to the polynucleotide of iii).   
     
     
         34 . The immunogenic composition according to  claim 32 , wherein said antigenic protein is selected from the group consisting of:
 i) a polypeptide comprising the sequence of SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:55, SEQ ID NO:58, or SEQ ID NO:123;   ii) a polypeptide that is at least 90% homologous to the polypeptide of i);   iii) a polypeptide that is encoded by a DNA comprising the sequence of SEQ ID NO:15, SEQ ID NO:19, or SEQ ID NO:25; and   iv) a polypeptide that is encoded by a polynucleotide that is at least 90% homologous to the polynucleotide of iii).   
     
     
         35 . The immunogenic composition according to  claim 33  or  claim 34 , wherein said antigenic protein of PCVB is selected from the group consisting of an inactivated PCVB antigen, a live modified or attenuated PCVB, and combinations thereof. 
     
     
         36 . The immunogenic composition according to  claim 33  or  claim 34 , wherein said antigenic protein of PCVB is expressed by recombinant baculovirus. 
     
     
         37 . The immunogenic composition according to  claim 33  or  claim 34 , wherein said further antigen from another disease-causing organism in swine is selected from the group consisting of an antigen of:  Actinobacillus pleuropneumonia ; Adenovirus; Alphavirus such as Eastern equine encephalomyelitis viruses;  Bordetella bronchiseptica; Brachyspira  spp., preferably  B. hyodyentheria; B. piosicoli, Brucella suis , preferably biovars 1, 2, and 3: Clasical swine fever virus;  Clostridium  spp., preferably  Cl. difficile, Cl. perfringens  types A, B, and C,  Cl. novyi, Cl. septicum, Cl. tetani ; Coronavirus, preferably Porcine Respiratory Corona virus; Eperythrozoonosis suis; Erysipelothrix rhsiopathiae;  Escherichia coli; Haemophilus parasuis , preferably subtypes 1, 7 and 14; Hemagglutinating encephalomyelitis virus;  Japanese Encephalitis Virus;  Lawsonia intracellularis; Leptospira  spp., preferably  Leptospira australis; Leptospira canicola; Leptospira grippotyphosa; Leptospira icterohaemorrhagicae ; and  Leptospira interrogans; Leptospira pomona; Leptospira tarassovi; Mycobacterium  spp., preferably  M. avium, M. intracellulare  and  M. bovis; Mycoplasma hyopneumoniae  (M hyo);  Pasteurella multocida ; Porcine cytomegalovirus; Porcine Parvovirus; Porcine Reproductive and Respiratory Syndrome (PRRS) Virus, Pseudorabies virus; Rotavirus;  Salmonella  spp., preferably  S. thyhimurium  and  S. choleraesuis; Staph. hyicus; Staphylococcus  spp., preferably  Streptococcus  spp., preferably  Strep. suis ; Swine herpes virus; Swine Influenza Virus;  Swine pox virus; Swine pox virus. Vesicular stomatitis virus; Virus of vesicular exanthema of swine, and combinations thereof 
     
     
         38 . A multivalent vaccine comprising:
 i) an antigenic protein of PCVB;   ii) at least one further antigen from another disease-causing organism in swine; and   iii) an adjuvant.   
     
     
         39 . The multivalent vaccine according to  claim 38 , wherein said adjuvant is aluminum hydroxide, a muramyl peptide, a bacterial lysate, Freund's incomplete adjuvant, AIF™, or AIF SEPPIC. 
     
     
         40 . The multivalent vaccine according to  claim 38  or  claim 39 , wherein said antigenic protein of PCVB reduces the incidence of tissues positive for the signs of PCVB infection in comparison to those tissues in an animal not receiving said immunogenic composition as determined by immunohistochemical testing for at least 25 days after the administration of a single dose of said multivalent vaccine. 
     
     
         41 . The multivalent vaccine according to  claim 38  or  claim 39 , wherein said antigenic protein of PCVB reduces the incidence of or lessens the severity of PCVB infections in comparison to those tissues in an animal not receiving said immunogenic composition for at least 25 days after the administration of a single dose of said multivalent vaccine. 
     
     
         42 . The multivalent vaccine according to  claim 38  or  claim 39 , wherein said antigenic protein of PCVB is selected from the group consisting of:
 i) a polypeptide comprising the sequence of SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:55, SEQ ID NO:58, or SEQ ID NO:123;   ii) a polypeptide that is at least 90% homologous to the polypeptide of i);   iii) a polypeptide that is encoded by a DNA comprising the sequence of SEQ ID NO:15, SEQ ID NO:19, or SEQ ID NO:25; and   iv) a polypeptide that is encoded by a polynucleotide that is at least 90% homologous to the polynucleotide of iii).   
     
     
         43 . The multivalent vaccine according to  claim 42 , wherein said antigenic protein of PCVB is included in inactivated PCVB antigen or any live modified or attenuated PCVB. 
     
     
         44 . The multivalent vaccine according to  claim 42 , wherein said antigenic protein of PCVB is expressed by recombinant baculovirus. 
     
     
         45 . The multivalent vaccine according to  claim 42 , wherein said further antigen from another disease-causing organism in swine is selected from the group consisting of an antigen of:  Actinobacillus pleuropneumonia ; Adenovirus; Alphavirus such as Eastern equine encephalomyelitis viruses;  Bordetella bronchiseptica; Brachyspira  spp., preferably  B. hyodyentheria; B. piosicoli, Brucella suis , preferably biovars 1, 2, and 3: Clasical swine fever virus;  Clostridium  spp., preferably  Cl. difficile, Cl. perfringens  types A, B, and C,  Cl. novyi, Cl. septicum, Cl. tetani ; Coronavirus, preferably Porcine Respiratory Corona virus; Eperythrozoonosis suis; Erysipelothrix rhsiopathiae;  Escherichia coli; Haemophilus parasuis , preferably subtypes 1, 7 and 14; Hemagglutinating encephalomyelitis virus; Japanese Encephalitis Virus;  Lawsonia intracellularis; Leptospira  spp., preferably  Leptospira australis; Leptospira canicola; Leptospira grippotyphosa; Leptospira icterohaemorrhagicae ; and  Leptospira interrogans; Leptospira pomona; Leptospira tarassovi; Mycobacterium  spp., preferably  M. avium, M. intracellulare  and  M. bovis; Mycoplasma hyopneumoniae  (M hyo);  Pasteurella multocida ; Porcine cytomegalovirus; Porcine Parvovirus; Porcine Reproductive and Respiratory Syndrome (PRRS) Virus, Pseudorabies virus; Rotavirus;  Salmonella  spp., preferably  S. thyhimurium  and  S. choleraesuis; Staph. hyicus; Staphylococcus  spp., preferably  Streptococcus  spp., preferably  Strep. suis ; Swine herpes virus; Swine Influenza Virus; Swine pox virus; Swine pox virus. Vesicular stomatitis virus; Virus of vesicular exanthema of swine, and combinations thereof. 
     
     
         46 . A multivalent immunogenic composition comprising:
 an antigenic protein of PCVB; and   at least one further antigen from another disease-causing organism in swine.   
     
     
         47 . The multivalent immunogenic composition according to  claim 46 , further comprising an adjuvant. 
     
     
         48 . The multivalent immunogenic composition according to  claim 46  or 47, wherein said antigenic protein of PCVB is ORF′2 (SEQ ID NO:26). 
     
     
         49 . The multivalent immunogenic composition according to  claim 46  or  47 , wherein said antigenic protein of PCVB is encoded by a polynucleotide sequence having at least 80% identity to the polynucleotide sequence encoding ORF′2 (SEQ ID NO:25). 
     
     
         50 . A multivalent vaccine comprising:
 an antigenic protein of PCVB; and   at least one further antigen from another disease-causing organism in swine.   
     
     
         51 . The multivalent vaccine according to  claim 50 , further comprising an adjuvant. 
     
     
         52 . The multivalent vaccine according to  claim 50  or  51 , wherein said antigenic protein of PCVB is ORF′2 (SEQ ID NO:26). 
     
     
         53 . The multivalent vaccine according to  claim 50  or  51 , wherein said antigenic protein of PCVB is encoded by a polynucleotide sequence having at least 80% identity to the polynucleotide sequence encoding ORF′2 (SEQ ID NO:25). 
     
     
         54 . A method for lessening and/or reduction of lymphadenopathy in combination with at least one symptom selected from the group consisting of: (1) interstitial pneumonia with interlobular edema, (2) cutaneous pallor or icterus, (3) mottled atrophic livers, (4) gastric ulcers, (5) nephritis and (6) reproductive disorders, e.g. abortion, stillbirths, mummies in pigs, comprising administering to a pig at least one dose of an immunogenic composition of at least 2 μg of recombinant PCVB ORF′2 protein. 
     
     
         55 . A method for reducing or lessening lymphadenopathy, lymphoid depletion and/or multinucleated/giant histiocytes in pigs infected with PCVB, comprising administering to a pig at least one dose of an immunogenic composition of at least 2 μg of recombinant PCVB ORF′2 protein. 
     
     
         56 . A method for reducing the overall circovirus load in a pig, comprising administering to that pig at least one dose of an immunogenic composition containing at least 2 μg of recombinant PCVB ORF′2 protein. 
     
     
         57 . A method for reducing the incidence of PCVB associated diseases and symptoms in pigs comprising administering to said pig at least one dose of an immunogenic composition containing at least 2 μg of recombinant PCVB ORF′2 protein. 
     
     
         58 . A method for the prevention, lessening the severity of and/or reduction of Pia like lesions, normally known to be associated with  Lawsonia intracellularis  infections in swine, comprising administering to a swine at least one dose of an immunogenic composition of at least 2 μg of recombinant PCVB ORF′2 protein. 
     
     
         59 . A method for reducing the immunosuppressive effect of porcine circovirus infection in pigs, comprising administering to a pig at least one dose of an immunogenic composition containing at least 2 μg of recombinant PCVB ORF′2 protein. 
     
     
         60 . A method of reducing or lessening the severity of clinical symptoms associated with porcine circovirus type B (PCVB) infection or lessening overall porcine circovirus load of an animal, said method selected from the group consisting of:
 (a) a method for lessening and/or reducing lymphadenopathy in combination with at least one symptom selected from the group consisting of: (1) interstitial pneumonia with interlobular edema, (2) cutaneous pallor or icterus, (3) mottled atrophic livers, (4) gastric ulcers, (5) nephritis and (6) reproductive disorders in pigs;   (b) a method for reducing or lessening lymphadenopathy, lymphoid depletion and/or multinucleated/giant histiocytes in pigs infected with PCVB;   (c) a method for reducing the overall circovirus load in a pig;   (d) a method for reducing the incidence of PCVB associated diseases and symptoms in pigs;   (e) a method for the prevention, lessening the severity of and/or reduction of Pia like lesions, normally known to be associated with  Lawsonia intracellularis  infections in swine; and   (f) a method for reducing the immunosuppressive effect of porcine circovirus infection in pigs;   comprising administering to a pig at least one dose of an immunogenic composition of at least 2 μg of recombinant PCVB ORF′2 protein.   
     
     
         61 . The method according to  claim 60 , wherein said PCVB ORF′2 protein is
 i) a polypeptide comprising the sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 55, SEQ ID NO: 58 and combinations thereof;   ii) any polypeptide that is at least 80% homologous to the polypeptide of i);   iii) any immunogenic portion of the polypeptides of i) and/or ii);   iv) the immunogenic portion of iii), comprising at least 10 contiguous amino acids included in a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 55, SEQ ID NO: 58 and combinations thereof;   v) a polypeptide that is encoded by a DNA comprising the sequence of SEQ ID NO: 15, SEQ ID NO: 19; SEQ ID NO: 23, SEQ ID NO: 25; SEQ ID NO: 27, or combinations thereof;   vi) any polypeptide that is encoded by a polynucleotide that is at least 80% homologous to the polynucleotide of v);   vii) any immunogenic portion of the polypeptides encoded by the polynucleotide of v) and/or vi); or   viii) the immunogenic portion of vii), wherein the polynucleotide coding for said immunogenic portion comprises at least 30 contiguous nucleotides included in the sequences of SEQ ID NO: 15, SEQ ID NO: 19; SEQ ID NO: 23, SEQ ID NO: 25; SEQ ID NO: 27, or combinations thereof.   
     
     
         62 . The method according to  claim 60 , wherein said composition further comprises an additional component selected from the group consisting of an inactivated viral vector, cell culture supernate, BEI, sodium thiosulfate, carriers, adjuvants, media, viral inactivators, diluents, isotonic agents, immunomodulatory agents, antibiotics, pharmaceutical acceptable salt, and combinations thereof. 
     
     
         63 . The method according to  claim 60 , wherein said inactivated viral vector is a recombinant baculovirus coding for the PCVB ORF′2 protein. 
     
     
         64 . The method according to  claim 60 , wherein the additional component comprises one or more adjuvants. 
     
     
         65 . The method according to  claim 60 , wherein one dose of said immunogenic composition comprising at least 4 μg of recombinant PCVB ORF′2 protein is administered to the pig. 
     
     
         66 . The method according to  claim 60 , wherein one dose of said immunogenic composition comprising at least 8 μg of recombinant PCVB ORF′2 protein is administered to the pig. 
     
     
         67 . The method according to  claim 60 , wherein one dose of said immunogenic composition comprising at least 16 μg of recombinant PCVB ORF′2 protein is administered to the pig. 
     
     
         68 . The method according to  claim 60 , wherein said immunogenic composition is effective for lessening the severity of clinical symptoms associated with PCVB infection after a single dose administration of said recombinant PCVB ORF′2 antigen. 
     
     
         69 . The method according to  claim 60 , wherein said immunogenic composition is administered to pigs of 2 weeks of age or older. 
     
     
         70 . The method according to  claim 60 , wherein said immunogenic composition is administered to pigs not older than 15 weeks of age. 
     
     
         71 . The method according to  claim 60 , wherein one dose of the immunogenic composition has a volume of 1 ml. 
     
     
         72 . The method according to  claim 60 , wherein said one dose of the immunogenic composition is administered to said pig in 1 ml. 
     
     
         73 . The method according to  claim 60 , wherein said one dose of the immunogenic composition is retained in a container. 
     
     
         74 . The method according to  claim 60 , wherein said immunogenic composition is effective for lessening the severity of clinical symptoms associated with PCVB infection after a single dose administration of said recombinant PCVB ORF′2 protein. 
     
     
         75 . A multivalent combination vaccine comprising an immunogenic composition that comprises at least 2 μg recombinant PCVB ORF′2 protein and at least one immunogenic active component against another disease-causing organism in swine, wherein said immunogenic composition is effective for lessening the severity of clinical symptoms associated with PCVB infection. 
     
     
         76 . The multivalent combination vaccine of  claim 75 , wherein said PCVB ORF′2 protein is effective for lessening the severity of clinical symptoms associated with PCVB infection after a single dose administration of said recombinant PCVB ORF′2 protein. 
     
     
         77 . The multivalent combination vaccine of  claim 75 , wherein said PCVB ORF′2 protein is
 i) a polypeptide comprising a sequence selected from the group consisting of SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:55, and SEQ ID NO:58;   ii) any polypeptide that is at least 90% homologous to the polypeptide of i);   iii) any immunogenic portion of the polypeptides of i) and/or ii);   iv) the immunogenic portion of iii), comprising at least 10 contiguous amino acids included in a sequence selected from the group consisting of SEQ ED NO:26, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:55, and SEQ ID NO:58;   v) a polypeptide that is encoded by a DNA comprising the sequence of SEQ ID NO:15, SEQ ID NO:19, or SEQ ID NO: 25;   vi) any polypeptide that is encoded by a polynucleotide that is at least 90% homologous to the polynucleotide of v);   vii) any immunogenic portion of the polypeptides encoded by the polynucleotide of v) and/or vi); or   viii) the immunogenic portion of vii), wherein polynucleotide coding for said immunogenic portion comprises at least 30 contiguous nucleotides included in the sequences of SEQ ID NO:15, SEQ ID NO:19, or SEQ ID NO: 25.   
     
     
         78 . The multivalent combination vaccine of  claim 75 , wherein said immunogenic composition that comprises recombinant PCVB ORF′2 protein further comprises an additional component selected from the group consisting of an inactivated viral vector, cell culture supernate, BEI, sodium thiosulfate, carriers, adjuvants, media, viral inactivators, diluents, isotonic agents, immunomodulators, agents, antibiotics, pharmaceutical acceptable salt, and combinations thereof. 
     
     
         79 . The multivalent combination vaccine of  claim 75 , wherein said immunogenic composition comprises at least 4 μg recombinant PCVB ORF′2 protein. 
     
     
         80 . The multivalent combination vaccine of  claim 75 , wherein one dose of said immunogenic composition has a volume of at least 1 ml. 
     
     
         81 . The multivalent combination vaccine of  claim 75 , wherein one dose of said immunogenic composition has a volume of at least 2 ml. 
     
     
         82 . The multivalent combination vaccine of  claim 75 , wherein said immunogenic composition is retained within a container. 
     
     
         83 . The multivalent combination vaccine of  claim 75 , wherein said further immunogenic active component is selected from the group consisting of  Mycoplasma hyopneumoniae , Porcine Reproductive and Respiratory Syndrome Virus, and combinations thereof. 
     
     
         84 . The multivalent combination vaccine of  claim 83 , wherein said further immunogenic active component is  Mycoplasma hyopneumoniae.    
     
     
         85 . The multivalent combination vaccine of  claim 83 , wherein said further immunogenic active component is Porcine Reproductive and Respiratory Syndrome Virus. 
     
     
         86 . The multivalent combination vaccine of  claim 83 , wherein said further immunogenic active components are  Mycoplasma hyopneumoniae  and Porcine Reproductive and Respiratory Syndrome Virus. 
     
     
         87 . The multivalent combination vaccine of  claim 83 , wherein said PCVB ORF′2 protein comprising immunogenic composition is effective for lessening the severity of clinical symptoms associated with PCVB infection after a single dose administration of said recombinant PCVB ORF′2 antigen. 
     
     
         88 . The multivalent combination vaccine of  claim 83 , wherein said combination vaccine comprises 3-10 logs of Porcine Reproductive and Respiratory Syndrome Virus. 
     
     
         89 . The multivalent combination vaccine of  claim 83 , wherein said immunogenic composition further comprises an additional component selected from the group consisting of an inactivated viral vector, cell culture supernate, BEI, sodium thiosulfate, carriers, adjuvants, media, viral inactivators, diluents, isotonic agents, immunomodulatory agents, antibiotics, pharmaceutical acceptable salt, and combinations thereof 
     
     
         90 . The multivalent combination vaccine of  claim 83 , wherein one dose of said immunogenic composition has a volume of about 1 ml. 
     
     
         91 . The multivalent combination vaccine of  claim 83 , wherein one dose of said immunogenic composition has a volume of about 2 ml. 
     
     
         92 . The multivalent combination vaccine of  claim 86 , wherein one dose of said immunogenic composition has a volume of about 2 ml.