{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,15]],"date-time":"2025-12-15T13:40:31Z","timestamp":1765806031252,"version":"build-2065373602"},"reference-count":25,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,2,19]],"date-time":"2022-02-19T00:00:00Z","timestamp":1645228800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Life"],"abstract":"<jats:p>In this study, we cultured the Bacillus anthracis vaccine strain Sterne 34F2 in a medium containing EDTA, and we assessed the best conditions to inhibit the activity of zinc-dependent metalloproteases to obtain a secretome containing a high concentration of non-degraded PA (PA83), as evaluated by the SDS-PAGE analysis. Then, we used this secretome as the antigen in a Complement Fixation Test (CFT) to monitor the production of antibodies against PA83 in the sera of rabbits vaccinated with Sterne 34F2 and then infected with a B. anthracis virulent strain to evaluate the potency of the vaccine. The PAS-based CFT results were compared with those obtained by using a commercial ELISA kit. The two serological tests gave similar results in terms of specificity and sensitivity, as the kinetics of the antibodies production was very similar. The Sterne 34F2 vaccine induced an antibody response to PA83, whose titer was not inferior to 1:8 in PAS-based CFT and 42 kU\/mL in PA83-based ELISA, respectively, in all vaccinated rabbits. Our opinion is that the PAS-based CFT can be successfully employed in humans and in animals for epidemiological retrospective studies or post-vaccination monitoring. We also suggest the use of our method to test the efficacy of veterinary anthrax vaccines.<\/jats:p>","DOI":"10.3390\/life12020312","type":"journal-article","created":{"date-parts":[[2022,2,21]],"date-time":"2022-02-21T08:22:19Z","timestamp":1645431739000},"page":"312","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Production of a Bacillus anthracis Secretome with Suitable Characteristics as Antigen in a Complement Fixation Test"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1159-2801","authenticated-orcid":false,"given":"Domenico","family":"Galante","sequence":"first","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Anthrax Reference Institute of Italy, Via Manfredonia 20, 71121 Foggia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8342-5375","authenticated-orcid":false,"given":"Viviana","family":"Manzulli","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Anthrax Reference Institute of Italy, Via Manfredonia 20, 71121 Foggia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adelia","family":"Donatiello","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Anthrax Reference Institute of Italy, Via Manfredonia 20, 71121 Foggia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonio","family":"Fasanella","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Anthrax Reference Institute of Italy, Via Manfredonia 20, 71121 Foggia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8403-1829","authenticated-orcid":false,"given":"Barbara","family":"Chirullo","sequence":"additional","affiliation":[{"name":"Department of Food Safety, Nutrition and Veterinary Public Health, Istituto Superiore di Sanit\u00e0, Viale Regina Elena 299, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Massimiliano","family":"Francia","sequence":"additional","affiliation":[{"name":"Department of Food Safety, Nutrition and Veterinary Public Health, Istituto Superiore di Sanit\u00e0, Viale Regina Elena 299, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7842-8697","authenticated-orcid":false,"given":"Valeria","family":"Rondinone","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Anthrax Reference Institute of Italy, Via 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Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5241-8044","authenticated-orcid":false,"given":"Michela","family":"Tarantino","sequence":"additional","affiliation":[{"name":"Department of Food Safety, Nutrition and Veterinary Public Health, Istituto Superiore di Sanit\u00e0, Viale Regina Elena 299, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rosanna","family":"Adone","sequence":"additional","affiliation":[{"name":"Department of Food Safety, Nutrition and Veterinary Public Health, Istituto Superiore di Sanit\u00e0, Viale Regina Elena 299, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3684","DOI":"10.1002\/pmic.200401218","article-title":"The extracellular and cytoplasmic proteomes of the non-virulent Bacillus anthracis strain UM23C1-2","volume":"5","author":"Antelmann","year":"2005","journal-title":"Proteomics"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1016\/j.mib.2006.02.009","article-title":"Regulatory networks for virulence and persistence of Bacillus anthracis","volume":"9","author":"Fouet","year":"2006","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1016\/j.mam.2009.07.004","article-title":"Bacillus anthracis physiology and genetics","volume":"30","author":"Koehler","year":"2009","journal-title":"Mol. Aspects Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1146\/annurev.cellbio.19.111301.140655","article-title":"Anthrax toxin","volume":"19","author":"Collier","year":"2003","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1146\/annurev.biochem.75.103004.142728","article-title":"Anthrax toxin: Receptor binding, internalization, pore formation, and translocation","volume":"76","author":"Young","year":"2007","journal-title":"Annu. Rev. Biochem."},{"key":"ref_6","unstructured":"World Health Organization (WHO) (2008). Anthrax in Animal and Humans, 4th ed, World Health Organization. Available online: http:\/\/www.who.int\/csr\/resources\/publications\/anthrax_webs.pdf."},{"key":"ref_7","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_8","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"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"4768","DOI":"10.1016\/S0264-410X(01)00234-1","article-title":"In vitro correlate of immunity in a rabbit model of inhalational anthrax","volume":"19","author":"Pitt","year":"2001","journal-title":"Vaccine"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"4409","DOI":"10.1016\/S0264-410X(01)00197-9","article-title":"Passive transfer of protection against Bacillus anthracis infection in a murine model","volume":"19","author":"Beedham","year":"2001","journal-title":"Vaccine"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2888","DOI":"10.1128\/IAI.69.5.2888-2893.2001","article-title":"Search of correlates of protective immunity conferred by anthrax vaccine","volume":"69","author":"Reuveny","year":"2001","journal-title":"Infect. Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"544","DOI":"10.1128\/IAI.70.2.544-550.2002","article-title":"Efficiency of protection of guinea pigs against infection with Bacillus anthracis spores by passive immunization","volume":"70","author":"Kobiler","year":"2002","journal-title":"Infect. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1077","DOI":"10.1515\/BC.2011.208","article-title":"Cysteine proteinase SpeB from Streptococcus pyogenes a potent modifier of immunologically important host and bacterial proteins","volume":"392","author":"Nelson","year":"2011","journal-title":"Biol. Chem."},{"key":"ref_14","first-page":"31408","article-title":"Secreted neutral metalloproteases of Bacillus anthracis as candidate pathogenic factors","volume":"281","author":"Chung","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1128\/JB.00690-13","article-title":"Modulation of the Bacillus anthracis secretome by the immune inhibitor A1 protease","volume":"196","author":"Pflughoeft","year":"2014","journal-title":"J. Bacteriol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fmicb.2016.00019","article-title":"Development of a Sterne-based complement fixation test to monitor the humoral response induced by anthrax vaccines","volume":"7","author":"Adone","year":"2016","journal-title":"Front. Microbiol."},{"key":"ref_17","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. Sci. Anim. Ind."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"4214","DOI":"10.1016\/S0264-410X(01)00159-1","article-title":"Detection of anthrax vaccine virulence factors by polymerase chain reaction","volume":"19","author":"Fasanella","year":"2001","journal-title":"Vaccine"},{"key":"ref_19","unstructured":"EDQM Council of Europe (2017). European Pharmacopoeia, EDQM Council of Europe. [9th ed.]."},{"key":"ref_20","unstructured":"OIE Manual (2016). Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, OIE, World Organisation for Animal Health. Part 2, Chapter 2.1.4."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1111\/j.1365-2672.2008.03787.x","article-title":"Brucella melitensis B115-based complement fixation test to detect antibodies induced by Brucella rough strains","volume":"10","author":"Adone","year":"2008","journal-title":"J. Appl. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1016\/j.devcel.2008.08.002","article-title":"Regulated secretion of a protease activates intercellular signaling during fruiting body formation in M. xanthus","volume":"15","author":"Rolbetzki","year":"2008","journal-title":"Dev. Cell."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"764","DOI":"10.1016\/j.wasman.2009.12.008","article-title":"Functional changes in culturable microbial communities during a co-composting process: Carbon source utilization and co-metabolism","volume":"30","author":"Albrecht","year":"2010","journal-title":"Waste Manag."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2131","DOI":"10.1128\/JB.00143-12","article-title":"Working together for the common good: Cell-cell communication in bacteria","volume":"194","author":"Stevens","year":"2012","journal-title":"J. Bacteriol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/S1369-5274(03)00003-1","article-title":"IdeS and SpeB: Immunoglobulin-degrading cysteine proteinases of Streptococcus pyogenes","volume":"6","author":"Bjorck","year":"2003","journal-title":"Curr. Opin. Microbiol."}],"container-title":["Life"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-1729\/12\/2\/312\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:23:01Z","timestamp":1760134981000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-1729\/12\/2\/312"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,19]]},"references-count":25,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["life12020312"],"URL":"https:\/\/doi.org\/10.3390\/life12020312","relation":{},"ISSN":["2075-1729"],"issn-type":[{"type":"electronic","value":"2075-1729"}],"subject":[],"published":{"date-parts":[[2022,2,19]]}}}