{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,8]],"date-time":"2026-02-08T22:40:08Z","timestamp":1770590408269,"version":"3.49.0"},"reference-count":50,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2020,7,10]],"date-time":"2020-07-10T00:00:00Z","timestamp":1594339200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Technology Innovation Agency (TIA)","award":["90052"],"award-info":[{"award-number":["90052"]}]},{"name":"National Research Foundation (NRF)","award":["98651"],"award-info":[{"award-number":["98651"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>The Sterne live spore vaccine (SLSV, Bacillus anthracis strain 34F2) is the veterinary vaccine of choice against anthrax though contra-indicated for use with antimicrobials. However, the use of non-living anthrax vaccine (NLAV) candidates can overcome the SLSV limitation. In this study, cattle were vaccinated with either of the NLAV (purified recombinant PA (PrPA) or crude rPA (CrPA) and formaldehyde-inactivated spores (FIS of B. anthracis strain 34F2) and emulsigen-D\u00ae\/alhydrogel\u00ae adjuvants) or SLSV. The immunogenicity of the NLAV and SLSV was assessed and the protective efficacies evaluated using a passive immunization mouse model. Polyclonal IgG (including the IgG1 subset) and IgM responses increased significantly across all vaccination groups after the first vaccination. Individual IgG subsets titres peaked significantly with all vaccines used after the second vaccination at week 5 and remained significant at week 12 when compared to week 0. The toxin neutralization (TNA) titres of the NLAV vaccinated cattle groups showed similar trends to those observed with the ELISA titres, except that the former were lower, but still significant, when compared to week 0. The opsonophagocytic assay indicated good antibody opsonizing responses with 75% (PrPA+FIS), 66% (CrPA+FIS) and 80% (SLSV) phagocytosis following spores opsonization. In the passive protection test, A\/J mice transfused with purified IgG from cattle vaccinated with PrPA+FIS+Emulsigen-D\u00ae\/Alhydrogel\u00ae and SLSV had 73% and 75% protection from challenge with B. anthracis strain 34F2 spores, respectively, whereas IgG from cattle vaccinated with CrPA+FIS+Emulsigen-D\u00ae\/Alhydrogel\u00ae offered insignificant protection of 20%. There was no difference in protective immune response in cattle vaccinated twice with either the PrPA+FIS or SLSV. Moreover, PrPA+FIS did not show any residual side effects in vaccinated cattle. These results suggest that the immunogenicity and protective efficacy induced by the NLAV (PrPA+FIS) in the cattle and passive mouse protection test, respectively, are comparable to that induced by the standard SLSV.<\/jats:p>","DOI":"10.3390\/pathogens9070557","type":"journal-article","created":{"date-parts":[[2020,7,10]],"date-time":"2020-07-10T09:25:28Z","timestamp":1594373128000},"page":"557","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Immunogenicity of Non-Living Anthrax Vaccine Candidates in Cattle and Protective Efficacy of Immune Sera in A\/J Mouse Model Compared to the Sterne Live Spore Vaccine"],"prefix":"10.3390","volume":"9","author":[{"given":"Solomon","family":"Jauro","sequence":"first","affiliation":[{"name":"Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Onderstepoort, Pretoria 0110, South Africa"},{"name":"Department of Veterinary Microbiology, Faculty of Veterinary Medicine, University of Maiduguri, Maiduguri 600230, Nigeria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2958-7151","authenticated-orcid":false,"given":"Okechukwu","family":"C. Ndumnego","sequence":"additional","affiliation":[{"name":"Moredun Scientific, Pentlands Science Park, Edinburgh EH26 0PZ, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Charlotte","family":"Ellis","sequence":"additional","affiliation":[{"name":"Design Biologix, Building 43b CSIR, Meiring Naude Road, Brummeria 0184, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angela","family":"Buys","sequence":"additional","affiliation":[{"name":"Design Biologix, Building 43b CSIR, Meiring Naude Road, Brummeria 0184, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6376-4230","authenticated-orcid":false,"given":"Wolfgang","family":"Beyer","sequence":"additional","affiliation":[{"name":"Department of Livestock Infectiology and Environmental Hygiene, Institute of Animal Science, University of Hohenheim, Stuttgart 70599, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3577-1273","authenticated-orcid":false,"given":"Henriette","family":"van Heerden","sequence":"additional","affiliation":[{"name":"Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Onderstepoort, Pretoria 0110, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,7,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/0264-410X(84)90003-3","article-title":"Anthrax: The disease in relation to vaccines","volume":"2","author":"Hambleton","year":"1984","journal-title":"Vaccine"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1016\/j.mam.2009.08.004","article-title":"Anthrax in animals","volume":"30","author":"Beyer","year":"2009","journal-title":"Mol. Asp. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1380","DOI":"10.1099\/jmm.0.045393-0","article-title":"Characterization of a multi-component anthrax vaccine designed to target the initial stages of infection as well as toxaemia","volume":"61","author":"Cote","year":"2012","journal-title":"J. Med. Microbiol."},{"key":"ref_4","first-page":"143","article-title":"Bacillus anthracis genetics and virulence gene regulation, in Anthrax","volume":"271","author":"Koehler","year":"2002","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1046\/j.1365-2958.1999.01137.x","article-title":"Germination of Bacillus anthracis spores within alveolar macrophages","volume":"31","author":"Mock","year":"1999","journal-title":"Mol. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"16409","DOI":"10.1073\/pnas.0508259102","article-title":"A model of anthrax toxin lethal factor bound to protective antigen","volume":"102","author":"Lacy","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3162","DOI":"10.1073\/pnas.79.10.3162","article-title":"Anthrax toxin edema factor: A bacterial adenylate cyclase that increases cyclic AMP concentrations of eukaryotic cells","volume":"79","author":"Leppla","year":"1982","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"7045","DOI":"10.1073\/pnas.052160199","article-title":"Biological Sciences\u2014Microbiology\u2014The lethal and edema factors of anthrax toxin bind only to oligomeric forms of the protective antigen","volume":"99","author":"Mogridge","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.3201\/eid1404.070969","article-title":"Conference report on public health and clinical guidelines for anthrax","volume":"14","author":"Stern","year":"2008","journal-title":"Emerg. Infect. Dis."},{"key":"ref_10","first-page":"307","article-title":"The use of anthrax vaccines prepared from avirulent (uncapsulated) variants of Bacillus anthracis","volume":"13","author":"Sterne","year":"1939","journal-title":"Onderstepoort J. Vet."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.vetimm.2016.06.005","article-title":"Comparative analysis of the immunologic response induced by the Sterne 34F2 live spore Bacillus anthracis vaccine in a ruminant model","volume":"178","author":"Ndumnego","year":"2016","journal-title":"Vet. Immunol. Immunopathol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"5684","DOI":"10.1016\/j.vaccine.2008.08.026","article-title":"Protective activity and immunogenicity of two recombinant anthrax vaccines for veterinary use","volume":"26","author":"Fasanella","year":"2008","journal-title":"Vaccine"},{"key":"ref_13","first-page":"25","article-title":"The immunization of laboratory animals against anthrax","volume":"13","author":"Sterne","year":"1939","journal-title":"Onderstepoort J. Vet."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1016\/0264-410X(91)90237-Z","article-title":"Anthrax vaccines: Past, present and future","volume":"9","author":"Turnbull","year":"1991","journal-title":"Vaccine"},{"key":"ref_15","unstructured":"Turnbull, P.C.B. (2008). Anthrax in Humans and Animals: Decisions on Treatment and\/or Vaccination of Livestock, WHO."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"16937","DOI":"10.1038\/s41598-018-35382-8","article-title":"Immunogenicity of anthrax recombinant peptides and killed spores in goats and protective efficacy of immune sera in A\/J mouse model","volume":"8","author":"Ndumnego","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1128\/IAI.70.2.661-664.2002","article-title":"Anthrax spores make an essential contribution to vaccine efficacy","volume":"70","author":"Brossier","year":"2002","journal-title":"Infect. Immun."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.ijmm.2003.12.002","article-title":"Protection of mice against challenge with Bacillus anthracis STI spores after DNA vaccination","volume":"294","author":"Hahn","year":"2004","journal-title":"Int. J. Med. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4569","DOI":"10.1016\/j.vaccine.2005.08.031","article-title":"DNA vaccination against anthrax in mice\u2014Combination of anti-spore and anti-toxin components","volume":"24","author":"Hahn","year":"2006","journal-title":"Vaccine"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1128\/iai.52.2.509-512.1986","article-title":"Comparative efficacy of Bacillus anthracis live spore vaccine and protective antigen vaccine against anthrax in the guinea pig","volume":"52","author":"Little","year":"1986","journal-title":"Infect. Immun."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4537","DOI":"10.1016\/j.vaccine.2009.05.045","article-title":"Comparability of ELISA and toxin neutralization to measure immunogenicity of Protective Antigen in mice, as part of a potency test for anthrax vaccines","volume":"27","author":"Parreiras","year":"2009","journal-title":"Vaccine"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3469","DOI":"10.1016\/j.vaccine.2006.02.013","article-title":"Comparative vaccine efficacy of different isoforms of recombinant protective antigen against Bacillus anthracis spore challenge in rabbits","volume":"24","author":"Ribot","year":"2006","journal-title":"Vaccine"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1016\/j.vaccine.2014.11.036","article-title":"Combination of two candidate subunit vaccine antigens elicits protective immunity to ricin and anthrax toxin in mice","volume":"33","author":"Vance","year":"2015","journal-title":"Vaccine"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"6379","DOI":"10.1016\/j.vaccine.2018.09.012","article-title":"Improving the stability of recombinant anthrax protective antigen vaccine","volume":"36","author":"Verma","year":"2018","journal-title":"Vaccine"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1186\/s12917-017-1140-2","article-title":"Protection of farm goats by combinations of recombinant peptides and formalin inactivated spores from a lethal Bacillus anthracis challenge under field conditions","volume":"13","author":"Koehler","year":"2017","journal-title":"BMC Vet. Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1096\/fj.09-138644","article-title":"Rabies DNA vaccine encoding lysosome-targeted glycoprotein supplemented with Emulsigen-D confers complete protection in preexposure and postexposure studies in BALB\/c mice","volume":"24","author":"Kaur","year":"2010","journal-title":"FASEB J."},{"key":"ref_27","unstructured":"McGonigle, J. (2006, January 16\u201319). Comparison of adjuvants for stimulation of HI antibody to SIV. Proceedings of the 19th IPVS Congress, Copenhage, Denmark."},{"key":"ref_28","first-page":"1","article-title":"Using Emulsigen-D as Recent Adjuvant in Trivalent Foot and Mouth Disease Vaccine","volume":"18","author":"Shabana","year":"2018","journal-title":"Glob. J. Med. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"492","DOI":"10.1016\/j.immuni.2010.10.002","article-title":"Vaccine adjuvants: Putting innate immunity to work","volume":"33","author":"Coffman","year":"2010","journal-title":"Immunity"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/S0169-409X(98)00008-8","article-title":"Aluminum compounds as vaccine adjuvants","volume":"32","author":"Gupta","year":"1998","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1038\/nri2510","article-title":"Towards an understanding of the adjuvant action of aluminium","volume":"9","author":"Marrack","year":"2009","journal-title":"Nat. Rev. Immunol."},{"key":"ref_32","first-page":"53","article-title":"The effect of large scale active immunization against anthrax","volume":"13","author":"Nicol","year":"1942","journal-title":"J. S. Afr. Vet. Assoc."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1111\/j.1751-0813.1973.tb02356.x","article-title":"Inhibiting effect of antibiotics on anthrax vaccination","volume":"49","author":"Webster","year":"1973","journal-title":"Aust. Vet. J."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Ascough, S., Ingram, R.J., Chu, K.K., Reynolds, C.J., Musson, J.A., Doganay, M., Metan, G., Metan, G., Ozkul, Y., and Baillie, L. (2014). Anthrax lethal factor as an immune target in humans and transgenic mice and the impact of HLA polymorphism on CD4+ T cell immunity. PLoS Pathog., 10.","DOI":"10.1371\/journal.ppat.1004085"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"5240","DOI":"10.1128\/IAI.00884-07","article-title":"Recombinant exosporium protein BclA of Bacillus anthracis is effective as a booster for mice primed with suboptimal amounts of protective antigen","volume":"75","author":"Brahmbhatt","year":"2007","journal-title":"Infect. Immun."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"5943","DOI":"10.4049\/jimmunol.0902795","article-title":"Contributions of edema factor and protective antigen to the induction of protective immunity by Bacillus anthracis edema toxin as an intranasal adjuvant","volume":"185","author":"Duverger","year":"2010","journal-title":"J. Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3103","DOI":"10.1099\/mic.0.28788-0","article-title":"Significant passive protective effect against anthrax by antibody to Bacillus anthracis inactivated spores that lack two virulence plasmids","volume":"152","author":"Enkhtuya","year":"2006","journal-title":"Microbiology"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.1128\/IAI.01217-08","article-title":"Efficacy of a vaccine based on protective antigen and killed spores against experimental inhalational anthrax","volume":"77","author":"Gauthier","year":"2009","journal-title":"Infect. Immun."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"7242","DOI":"10.1038\/s41598-018-25502-9","article-title":"A bivalent protein r-PB, comprising PA and BclA immunodominant regions for comprehensive protection against Bacillus anthracis","volume":"8","author":"Majumder","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"498","DOI":"10.3389\/fimmu.2019.00498","article-title":"A bivalent protein r-PAbxpB comprising PA domain IV and exosporium protein BxpB confers protection against B. anthracis spores and toxin","volume":"10","author":"Majumder","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"5171","DOI":"10.1128\/iai.65.12.5171-5175.1997","article-title":"Passive protection by polyclonal antibodies against Bacillus anthracis infection in guinea pigs","volume":"65","author":"Little","year":"1997","journal-title":"Infect. Immun."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1807","DOI":"10.1128\/iai.56.7.1807-1813.1988","article-title":"Production and characterization of monoclonal antibodies to the protective antigen component of Bacillus anthracis toxin","volume":"56","author":"Little","year":"1988","journal-title":"Infect. Immun."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1128\/CVI.00550-12","article-title":"Immunization of mice with formalin-inactivated spores from avirulent Bacillus cereus strains provides significant protection from challenge with Bacillus anthracis Ames","volume":"20","author":"Vergis","year":"2013","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/S0928-8244(03)00069-5","article-title":"Lipopeptides: Adjuvanticity in conventional and genetic immunization","volume":"37","author":"Heinevetter","year":"2003","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/0882-4010(88)90015-0","article-title":"Comparative safety and efficacy against Bacillus anthracis of protective antigen and live vaccines in mice","volume":"5","author":"Welkos","year":"1988","journal-title":"Microb. Pathog."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1086\/653495","article-title":"Select human anthrax protective antigen epitope-specific antibodies provide protection from lethal toxin challenge","volume":"202","author":"Crowe","year":"2010","journal-title":"J. Infect. Dis."},{"key":"ref_47","first-page":"28","article-title":"Antibody-based defense strategies against biological weapons","volume":"71","author":"Casadevall","year":"2005","journal-title":"ASM News"},{"key":"ref_48","unstructured":"OIE (2012). Manual of Diagnostic Tests and Vaccines for Terrestrial Animals (Mammals, Birds and Bees): Anthrax. Ch. 2.1.1, Office International des \u00c9pizooties."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"5978","DOI":"10.1128\/IAI.73.9.5978-5987.2005","article-title":"Immunogenicity of recombinant protective antigen and efficacy against aerosol challenge with anthrax","volume":"73","author":"Williamson","year":"2005","journal-title":"Infect. Immun."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1677","DOI":"10.1099\/00221287-147-6-1677","article-title":"The role of antibodies to Bacillus anthracis and anthrax toxin components in inhibiting the early stages of infection by anthrax spores","volume":"147","author":"Welkos","year":"2001","journal-title":"Microbiology"}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/9\/7\/557\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:49:47Z","timestamp":1760176187000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/9\/7\/557"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,10]]},"references-count":50,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2020,7]]}},"alternative-id":["pathogens9070557"],"URL":"https:\/\/doi.org\/10.3390\/pathogens9070557","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,7,10]]}}}