{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T07:41:08Z","timestamp":1773819668163,"version":"3.50.1"},"reference-count":49,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2025,1,14]],"date-time":"2025-01-14T00:00:00Z","timestamp":1736812800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Brucellosis is still the most common zoonosis worldwide despite advanced technology and animal husbandry. Since there is still no effective Brucella vaccine for humans, it is crucial to control the disease in ruminants through eradication and vaccination. Although some countries around the world have achieved this circumstance, every country aims to become free of Brucellosis through vaccination, animal movements, and various eradication measures. For this purpose, the Brucella abortus S19 strain has been used safely for about 100 years. However, due to the O-polysaccharide (OPS) antigen in its structure, the antibody response created by the vaccine causes confusion in serological tests. For this purpose, researchers have provided both mucosal immunity and short-term antibody response by using the B. abortus S19 vaccine in conjunctival form instead of subcutaneous form. This study aimed to determine how long the post-vaccination titer levels persisted in animals vaccinated with vaccines from 3 different companies and different routes. In this study, a total of 115 calves aged 3 to 4.5 months were created in five groups, with 23 animals in each group: group 1 (vaccine brand A), group 2 (vaccine brand B), and group 3 (vaccine brand C) received the two-dose conjunctival vaccine, group 4 received the single-dose subcutaneous vaccine (vaccine brand C), and group 5 received the subcutaneous vaccine (vaccine brand C) plus the booster dose conjunctival vaccine (vaccine brand B). Brucellosis antibody titers were monitored each 21 days until the cattle were 26\u201328 months old. The collected sera were analyzed using the Rose Bengal Plate Test (RBPT), Serum Agglutination Test (SAT), and Complement Fixation Test (CFT), which are the preferred serological methods for Brucellosis eradication plans worldwide. In the conjunctival vaccination groups, only 3 (13%) of the animals in group 1 developed antibody titers one month after vaccination, and there was no antibody response detected against Brucellosis in group 2 and group 3. In animals that were stimulated conjunctivally, the threshold value of 30 International CFT Units (ICFTUs) (for distinguishing between infective titers and vaccination titers) was observed in one animal each in group 1 and group 2 and 0 animal in group 3. It was found that antibody titers turn to Brucellosis negative in all conjunctival vaccine groups at 7 months after vaccination. In groups 4 and 5, the first-month serological screening detected over 30 ICFTUs in 17 (89.47%) animals and 16 (69.5%) animals, respectively. In group 4, CFT titers were found to fall below 30 on the 17th month and 9.3 on the 22nd month. On the 14th month, the CFT titers of group 5 were found to be below 30, and all animals in this group turned negative after the 19th month. It was found that the single dose B. abortus S19 subcutaneous vaccination in calves caused persistent antibodies in 5% of the population. It is believed that persistent and high antibody titers created by subcutaneous vaccines will cause false positivity and create confusion in Brucellosis eradication programs. Therefore, although there is no clear distinction between vaccinated and infected animals, it has been observed that conjunctival Brucellosis vaccines create more stable antibody titers and decrease rapidly compared to subcutaneous vaccines. Based on the results of this study and the advantages of conjunctival vaccines, more effective eradication programs and antibody monitoring can be carried out in vaccinated herds where Brucellosis outbreaks are observed.<\/jats:p>","DOI":"10.3390\/ani15020212","type":"journal-article","created":{"date-parts":[[2025,1,14]],"date-time":"2025-01-14T11:28:31Z","timestamp":1736854111000},"page":"212","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Comparison of Long-Term Antibody Titers in Calves Treated with Different Conjunctival and Subcutaneous Brucella abortus S19 Vaccines"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8319-831X","authenticated-orcid":false,"given":"Ali","family":"Uslu","sequence":"first","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine, Selcuk University, 42130 Konya, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zafer","family":"Say\u0131n","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine, Selcuk University, 42130 Konya, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Asl\u0131","family":"Balevi","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine, Selcuk University, 42130 Konya, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yasin","family":"Gulcu","sequence":"additional","affiliation":[{"name":"Konya Veterinary Control and Research Institute, 42080 Konya, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"F\u0131rat","family":"Ergen","sequence":"additional","affiliation":[{"name":"G\u00f6zl\u00fc Agricultural Enterprise, 42430 Konya, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Islam","family":"Ak\u0131ner","sequence":"additional","affiliation":[{"name":"Konya Veterinary Control and Research Institute, 42080 Konya, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oguzhan","family":"Denizli","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine, Selcuk University, 42130 Konya, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Osman","family":"Erganis","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Faculty of Veterinary Medicine, Selcuk University, 42130 Konya, Turkey"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,14]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2014, January 19\u201320). The control of neglected zoonotic diseases: From advocacy to action. Proceedings of the Fourth International Meeting held at WHO Headquarters, Geneva, Switzerland."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.micinf.2018.06.001","article-title":"Zoonoses under our noses","volume":"21","author":"Cross","year":"2019","journal-title":"Microbes Infect."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1080\/01652176.2020.1868616","article-title":"Bovine brucellosis\u2014A comprehensive review","volume":"41","author":"Khurana","year":"2021","journal-title":"Vet. Q."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"451","DOI":"10.2460\/javma.23.01.0033","article-title":"A one-health review on brucellosis in the United States","volume":"261","author":"Frye","year":"2023","journal-title":"J. Am. Vet. Med. Assoc."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"915","DOI":"10.1586\/14760584.4.6.915","article-title":"Essential role of vaccines in brucellosis control and eradication programs for livestock","volume":"4","author":"Olsen","year":"2005","journal-title":"Expert Rev. Vaccines"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"P\u00e9rez-Sancho, M., Garc\u00eda-Seco, T., Dom\u00ednguez, L., and \u00c1lvarez, J. (2015). Control of animal brucellosis, The most effective tool to prevent human brucellosis. Updates on Brucellosis, IntechOpen.","DOI":"10.5772\/61222"},{"key":"ref_7","first-page":"193","article-title":"The story of John M. Buck\u2019s and Matilda\u2019s contribution to the cattle industry","volume":"102","author":"Graves","year":"1943","journal-title":"J. Am. Vet. Med. Assoc."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1007\/s11515-012-1196-0","article-title":"Progress in Brucella vaccine development","volume":"8","author":"Yang","year":"2013","journal-title":"Front. Biol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1080\/14787210.2022.2066521","article-title":"Brucellosis control methods with an emphasis on vaccination: A systematic review","volume":"20","author":"Darbandi","year":"2022","journal-title":"Expert Rev. Anti-Infect. Ther."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Heidary, M., Dashtbin, S., Ghanavati, R., Mahdizade Ari, M., Bostanghadiri, N., Darbandi, A., Navidifar, T., and Talebi, M. (2022). Evaluation of brucellosis vaccines: A comprehensive review. Front. Vet. Sci., 9.","DOI":"10.3389\/fvets.2022.925773"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/bs.mim.2019.11.003","article-title":"Immunodetection of bacteria causing brucellosis","volume":"Volume 47","author":"Barbuddhe","year":"2020","journal-title":"Methods in Microbiology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1136\/vr.108.5.90","article-title":"Characterisation of Brucella abortus strain 19 cultures isolated from vaccinated cattle","volume":"108","author":"Thomas","year":"1981","journal-title":"Vet. Rec."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1016\/S0378-1135(02)00229-8","article-title":"Diagnosis of brucellosis by serology","volume":"90","author":"Nielsen","year":"2002","journal-title":"Vet. Microbiol."},{"key":"ref_14","first-page":"159","article-title":"Persistence of Brucella abortus, strain 19 infection in immunized cattle","volume":"73","author":"Meyer","year":"1969","journal-title":"Proc. Annu. Meet. U. S. Anim. Health Assoc."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Blasco, J.M., Moreno, E., Mu\u00f1oz, P., Conde-\u00c1lvarez, R., and Moriyon, I. (2023). A review of three decades of use of the cattle brucellosis rough vaccine Brucella abortus RB51: Myths and facts. BMC Vet. Res., 19.","DOI":"10.1186\/s12917-023-03773-3"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.prevetmed.2018.10.002","article-title":"Animal brucellosis control or eradication programs worldwide: A systematic review of experiences and lessons learned","volume":"160","author":"Zhang","year":"2018","journal-title":"Prev. Vet. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.cimid.2019.03.015","article-title":"Immunogenic and protective antigens of Brucella as vaccine candidates","volume":"65","author":"Jezi","year":"2019","journal-title":"Comp. Immunol. Microbiol. Infect. Dis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1186\/1297-9716-43-29","article-title":"What have we learned from brucellosis in the mouse model?","volume":"43","author":"Moreno","year":"2012","journal-title":"Vet. Res."},{"key":"ref_19","first-page":"44","article-title":"Brucella Eradication Programs and Country Models in the World","volume":"3","year":"2014","journal-title":"INFOVET"},{"key":"ref_20","first-page":"1","article-title":"The role of university-public-industry collaboration in the development of animal vaccines","volume":"26","year":"2010","journal-title":"Eurasian J. Vet. Sci."},{"key":"ref_21","unstructured":"(2024, June 12). Ministry of Agriculture and Forestry, Available online: https:\/\/www.tarimorman.gov.tr\/Konular\/Bitkisel-Uretim\/Iyi-Tarim-Uygulamalari\/Istatistikler."},{"key":"ref_22","first-page":"e32","article-title":"Efficacy of Brucella abortus S19 and RB51 vaccine strains: A systematic review and meta-analysis","volume":"69","author":"Pereira","year":"2022","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1006\/biol.2000.0249","article-title":"In vitro markers and biological activity in mice of seed lot strains and commercial Brucella melitensis Rev 1 and Brucella abortus B19 vaccines","volume":"28","author":"Bosseray","year":"2000","journal-title":"Biologicals"},{"key":"ref_24","unstructured":"(2024, July 24). OIE. Brucellosis (Brucella abortus, B. melitensis and B. suis) (infection with B. abortus, B. melitensis and B. suis). Available online: https:\/\/www.woah.org\/en\/what-we-do\/standards\/codes-and-manuals\/terrestrial-code-online-access\/."},{"key":"ref_25","unstructured":"Manthei, C.A. (1959, January 15\u201318). Summary of controlled research with Strain 19. Proceedings of the 63rd Meeting U.S. Livestock Sanitary Association, San Francisco, CA, USA."},{"key":"ref_26","first-page":"877","article-title":"The degree and duration of immunity in cattle resulting from vaccination with S. 19 Br. abortus vaccine and its implication in the future control and eventual eradication of brucellosis","volume":"69","author":"McDiarmid","year":"1957","journal-title":"Vet. Rec."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3109\/10408419009105726","article-title":"Pathogenesis of brucella","volume":"17","author":"Smith","year":"1990","journal-title":"Crit. Rev. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1016\/0007-1935(85)90046-6","article-title":"The effect of reduced-dose Brucella abortus strain 19 vaccination in accredited dairy herds","volume":"141","author":"Beckett","year":"1985","journal-title":"Br. Vet. J."},{"key":"ref_29","first-page":"203","article-title":"Stability of reduced virulence exhibited by Brucella abortus strain 19","volume":"99","author":"Jasmin","year":"1941","journal-title":"J. Am. Vet. Med. Assoc."},{"key":"ref_30","first-page":"317","article-title":"The stability of the avirulent characters of Brucella abortus, strain 19 and strain 45\/20 in lactating and pregnant cows","volume":"61","author":"Taylor","year":"1949","journal-title":"Vet. Rec."},{"key":"ref_31","first-page":"9","article-title":"Vaccination against bovine brucellosis with a low dose of strain 19 administered by the conjunctival route. III.-Serological response and immunity in the pregnant cow","volume":"7","author":"Plommet","year":"1976","journal-title":"Ann. Rech. Vet."},{"key":"ref_32","first-page":"345","article-title":"Vaccination against bovine brucellosis with a low dose of strain 19 administered by the conjunctival route. I.-protection demonstrated in guinea pigs","volume":"6","author":"Plommet","year":"1975","journal-title":"Ann. Rech. V\u00e9t\u00e9rinaires"},{"key":"ref_33","first-page":"1","article-title":"Vaccination against bovine brucellosis with a low dose of strain 19 administered by the conjunctival route. II.-determination of the minimum dose leading to colonization of the regional lymph nodes of cattle","volume":"7","author":"Plommet","year":"1976","journal-title":"Ann. Rech. Vet."},{"key":"ref_34","first-page":"131","article-title":"Vaccination against bovine brucellosis with a low dose of strain 19 administered by the conjunctival route. IV. Comparison between two methods of vaccination","volume":"10","author":"Fensterbank","year":"1979","journal-title":"Ann. Rech. V\u00e9t\u00e9rinaires"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/0264-410X(84)90004-5","article-title":"Vaccination of cattle with Brucella abortus strain 19 administered by differing routes and doses","volume":"2","author":"Nicoletti","year":"1984","journal-title":"Vaccine"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Dorneles, E.M., Lima, G.K., Teixeira-Carvalho, A., Ara\u00fajo, M.S., Martins-Filho, O.A., Sriranganathan, N., Al Qublan, H., Heinemann, M.B., and Lage, A.P. (2015). Immune response of calves vaccinated with Brucella abortus S19 or RB51 and revaccinated with RB51. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0136696"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1128\/CMR.16.1.65-78.2003","article-title":"Molecular host-pathogen interaction in brucellosis: Current understanding and future approaches to vaccine development for mice and humans","volume":"16","author":"Ko","year":"2003","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_38","first-page":"19","article-title":"Brucella abortus vaccine strain RB51: A brief review","volume":"27","author":"Schurig","year":"1995","journal-title":"Arch. Med. Vet."},{"key":"ref_39","first-page":"4994","article-title":"Assessment of listing and categorisation of animal diseases within the framework of the Animal Health Law (Regulation (EU) No 2016\/429): Ovine epididymitis (Brucella ovis)","volume":"15","author":"Velarde","year":"2017","journal-title":"EFSA J."},{"key":"ref_40","unstructured":"Robinson, A., and Production, A. (2003). Guidelines for Coordinated Human and Animal Brucellosis Surveillance, FAO."},{"key":"ref_41","first-page":"61","article-title":"Resposta sorol\u00f3gica de bovinos adultos ap\u00f3s vacina\u00e7\u00e3o com as amostras 19 e RB51 de Brucella abortus","volume":"37","author":"Poester","year":"2000","journal-title":"Braz. J. Vet. Res. Anim. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/0378-1135(82)90028-1","article-title":"Serological response of cattle after vaccination and challenge with Brucella abortus","volume":"7","author":"Sutherland","year":"1982","journal-title":"Vet. Microbiol."},{"key":"ref_43","first-page":"11","article-title":"The serological response of cattle to vaccines against brucellosis, as measured by the brucellosis radioimmunoassay and other tests","volume":"88","author":"Chappel","year":"1982","journal-title":"Epidemiol. Infect."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1007\/s11250-014-0678-2","article-title":"Vaccination of adult animals with a reduced dose of Brucella abortus S19 vaccine to control brucellosis on dairy farms in endemic areas of India","volume":"47","author":"Chand","year":"2015","journal-title":"Trop. Anim. Health Prod."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.4102\/jsava.v89i0.1527","article-title":"Immunological response to Brucella abortus strain 19 vaccination of cattle in a communal area in South Africa","volume":"89","author":"Simpson","year":"2018","journal-title":"J. S. Afr. Vet. Assoc."},{"key":"ref_46","first-page":"556","article-title":"Investigating Field Efficacy and Safety of Conjunctival Brucella abortus S19 Vaccine in Cattle","volume":"43","author":"Gurbilek","year":"2023","journal-title":"Agric. Sci. Dig."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1016\/0007-1935(87)90025-X","article-title":"Persistent serological titres following reduced dose Brucella abortus strain 19 vaccination","volume":"143","author":"Beckett","year":"1987","journal-title":"Br. Vet. J."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1007\/s11250-023-03519-1","article-title":"Evaluation of serological diagnostic tests for bovine brucellosis in dairy cattle herds in an endemic area: A multicenter study","volume":"55","author":"Alamian","year":"2023","journal-title":"Trop. Anim. Health Prod."},{"key":"ref_49","unstructured":"USDA (2024, November 12). NVAP Reference Guide: Brucellosis (Control and Eradication), Available online: https:\/\/www.aphis.usda.gov\/nvap\/reference-guide\/control-eradication\/brucellosis."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/2\/212\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T10:28:40Z","timestamp":1759919320000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/2\/212"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,14]]},"references-count":49,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2025,1]]}},"alternative-id":["ani15020212"],"URL":"https:\/\/doi.org\/10.3390\/ani15020212","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,1,14]]}}}