Disclosed is a gene multiple insertion cassette set including rDNA NTS fragments and an auxotrophic selection marker having an incomplete promoter is developed, and a safe oral recombinant strain having no antibiotic resistant marker is constructed by multiple insertion of an optimum number of the developed gene multiple insertion cassette sets into chromosomes of a Saccharomyces cerevisiae strain, a vaccine composition including, as an active ingredient, the above strain, a culture product thereof, a cell lysate, or nodavirus capsid protein (NNVcp) isolated and purified therefrom, and a composition for feed addition including, as an active ingredient, the above strain, a culture product thereof, a cell lysate, or squalene or oxidosqualene isolated and purified therefrom.

A nucleic acid constructs including a heterologous nucleotide sequence operably linked to a constitutively active promoter; a 5 recombination sequence at a 5 end of the construct and a 3 recombination sequence at a 3 end of the construct, wherein the recombination sequences are configured for homologous recombination into a genome of a microalgal host cell; and optionally a bacterial 5 untranslated region (5UTR) between the 5 recombination sequence and the promoter, and a bacterial 3 untranslated region (3UTR) between the 3 recombination sequence and the heterologous nucleotide sequence.

An oral vaccine composition for fish, and related oral vaccine delivery system and methods for making and using such oral vaccine composition and delivery system are described. The oral vaccine composition includes virus-like particles effective to mediate an immunoprotective response in a target fish species, wherein the virus-like particles are contained in biomass of an organism or cell culture in which the virus-like particles have been expressed, optionally further including (i) adjuvant effective to enhance the immunoprotective response of the virus-like particles in the target fish species; (ii) encapsulant protectively associated with the virus-like particles for selective locus bioavailability in the target fish species; and/or (iii) binding agent aggregating the ingredients of the composition with one another.

The present invention relates to stable compositions comprising acellular pertussis antigens that have not been cross-linked with a cross-linking agent such as formaldehyde or glutaraldehyde and their use as acellular pertussis components in combination vaccines. Processes for preparing these antigens and compositions are also disclosed.

The present invention provides edible vaccines comprising transgenic microalgae expressing at least one exogenous antigen or an intervening organism comprising the transgenic microalgae. The antigen expressing microalgae are used for oral delivery of the antigen to a target organism in its intact and functional form. The exogenous antigen, expressed in the microalgae, is characterized by exerting at least one immunogenic response in the subject consuming the vaccine.

Provided are edible vaccines including transgenic microalgae expressing at least one exogenous antigen or an intervening organism including the transgenic microalgae. The antigen expressing microalgae are used for oral delivery of the antigen to a target organism in its intact and functional form. The exogenous antigen, expressed in the microalgae, is characterized by exerting at least one immunogenic response in the subject consuming the vaccine.

Disclosed is a gene multiple insertion cassette set including rDNA NTS fragments and an auxotrophic selection marker having an incomplete promoter is developed, and a safe oral recombinant strain having no antibiotic resistant marker is constructed by multiple insertion of an optimum number of the developed gene multiple insertion cassette sets into chromosomes of a Saccharomyces cerevisiae strain, a vaccine composition including, as an active ingredient, the above strain, a culture product thereof, a cell lysate, or nodavirus capsid protein (NNVcp) isolated and purified therefrom, and a composition for feed addition including, as an active ingredient, the above strain, a culture product thereof, a cell lysate, or squalene or oxidosqualene isolated and purified therefrom.

Methods of producing, propagating and multiplying viruses, methods of identifying the presence of a virus in a sample, specifically methods using Japanese eel cell cultures. Also provided are an isolated Japanese eel cell culture, optionally infected with a virus, vaccines comprising viruses grown according to the methods of the invention, and a kit for propagating viruses comprising the Japanese eel cell culture. The virus may be a fish virus such as nodavirus, Infectious Pancreatic Necrosis Virus (IPNV) and Salmon Pancreas Disease Virus (SPDV).

An oral vaccine composition for fish, and related oral vaccine delivery system and methods for making and using such oral vaccine composition and delivery system are described. The oral vaccine composition includes virus-like particles effective to mediate an immunoprotective response in a target fish species, wherein the virus-like particles are contained in biomass of an organism or cell culture in which the virus-like particles have been expressed, optionally further including (i) adjuvant effective to enhance the immunoprotective response of the virus-like particles in the target fish species; (ii) encapsulant protectively associated with the virus-like particles for selective locus bioavailability in the target fish species; and/or (iii) binding agent aggregating the ingredients of the composition with one another.

A nucleic acid constructs including a heterologous nucleotide sequence operably linked to a constitutively active promoter; a 5 recombination sequence at a 5 end of the construct and a 3 recombination sequence at a 3 end of the construct, wherein the recombination sequences are configured for homologous recombination into a genome of a microalgal host cell; and optionally a bacterial 5 untranslated region (5UTR) between the 5 recombination sequence and the promoter, and a bacterial 3 untranslated region (3UTR) between the 3 recombination sequence and the heterologous nucleotide sequence.