Kansas State University Research Foundation

Tiruvoor G Nagaraja

Muckatira M Chengappa

Sanjeev K Narayanan

Amit Kumar

George C Stewart

The present invention relates to protecting against, treating, and detecting Fusobacteria infections. Compositions and methods derived from nucleic acid and protein sequences of a 40 kDa Adhesin protein are provided to protect against, treat, and detect Fusobacteria infections in a subject. In one aspect, vaccines capable of inducing an immune response to a 40 kDa Adhesin protein are used to protect against Fusobacteria infection. Also, nucleic acid molecules, proteins, immunogens, antibodies, and antisense molecules derived from the sequences of the 40 kDa Adhesin protein may be used to protect against, treat, and detect Fusobacteria infections in a subject.

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The present invention relates to protecting against, treating, and detectinginfections. Compositions and methods derived from nucleic acid and protein sequences of a 40 kDa Adhesin protein are provided to protect against, treat, and detectinfections in a subject. In one aspect, vaccines capable of inducing an immune response to a 40 kDa Adhesin protein are used to protect againstinfection. Also, nucleic acid molecules, proteins, immunogens, antibodies, and antisense molecules derived from the sequences of the 40 kDa Adhesin protein may be used to protect against, treat, and detectinfections in a subject.

Compositions and methods for detecting, treating, and protecting against fusobacterium infection

Theexpressing leukotoxin was sequenced and cloned. The leukotoxin open reading frame (lktA) is part of a multi-gene operon containing 9,726 bp, and encoding a protein containing 3,241 amino acids with an overall molecular weight of 335,956 daltons. The protein encoded by the gene was truncated into five polypeptides having overlapping regions by truncating the full length gene into five different sections and amplifying, expressing, and recovering the protein encoded by each of these sections. Additionally, a region upstream of the gene was sequenced and the polypeptide encoded by that nucleotide sequence was purified and isolated. These polypeptides along with the full length protein are then tested to determine their immunogenicity and protective immunity in comparison to the efficacy of immunization conferred by inactivated native leukotoxin inculture supernatant.

The gene expressing leukotoxin was sequenced and cloned. The leukotoxin open reading frame (lktA) is part of a multi-gene operon containing 9,726 bp, and encoding a protein containing 3,241 amino acids with an overall molecular weight of 335,956 daltons. The protein encoded by the gene was truncated into five polypeptides having overlapping regions by truncating the full length gene into five different sections and amplifying, expressing, and recovering the protein encoded by each of these sections. Additionally, a region upstream of the gene was sequenced and the polypeptide encoded by that nucleotide sequence was purified and isolated. These polypeptides along with the full length protein are then tested to determine their immunogenicity and protective immunity in comparison to the efficacy of immunization conferred by inactivated native leukotoxin in culture supernatant.

Recombinant fusobacterium necrophorum leukotoxin vaccine and preparation thereof

Novel inocula for administration to ruminant animals such as cattle or sheep are provided in order to immunize the animals and lessen the incidence of liver abscesses and/or foot rot therein. In one aspect, the invention pertains to an A. pyogenes-derived vaccine including an inactivated cell culture product (e.g., cell-elaborated supernatant) from A. pyogenes cell culture in a suitable carrier. In another aspect, the invention relates to a multivalent vaccine including at least first and second bacterial components in a carrier; the first component comprises an inactivated cell culture product of A. pyogenes whereas the second component comprises an inactivated cell culture product of F. necrophorum. The inocula of the present invention find particular utility in incidences where ruminant animals are particularly subject to A. pyogenes infection leading to liver abscesses and/or foot rot, e.g., where the animals are regularly treated with an antibiotic or where cattle are fed a high grain content concentrate diet.

Multivalent inocula for lessening incidence of liver abscesses in cattle

A method is provided for the enhanced elaboration of leukotoxin from F. necrophorum, and subsequent production of an inactivated leukotoxoid ruminant animal vaccine against F. necrophorum infection and consequent liver abscesses and/or foot rot in such animals. The method involves forming a culture of F. necrophorum bacteria in growth media, allowing the bacteria to grow therein and to simultaneously elaborate leukotoxin in a supernate; the culturing is preferably carried out at a temperature of from about 35.degree.-41.degree. C., a pH of from about 6.5-8, and for a period of from about 4-9 hours. At the end of the culturing, bacterial growth and leukotoxin elaboration are terminated, preferably by separating the leukotoxin supernate, whereupon the vaccine is produced by inactivation of at least the supernate.

Fusobacterium leukotoxoid vaccine

A method is provided for the enhanced elaboration of leukotoxin from F. necrophorum, and subsequent production of an inactivated leukotoxoid ruminant animal vaccine against F. necrophorum infection and consequent liver abscesses and/or foot rot in such animals. The method involves forming a culture of F. necrophorum bacteria in growth media, allowing the bacteria to grow therein and to simultaneously elaborate leukotoxin in a supernate; the culturing is preferably carried out at a temperature of from about 35.degree.-41.degree. C., a pH of from about 6.5-8, and for a period of from about 4-9 hours. At the end oil of the culturing, bacterial growth and leukotoxin elaboration are terminated, preferably by separating the leukotoxin supernate, whereupon the vaccine is produced by inactivation of at least the supernate.

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Manhattan, KS, United States of America

Tiruvoor G Nagaraja