{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T11:39:56Z","timestamp":1782819596596,"version":"3.54.5"},"reference-count":27,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2013,3,21]],"date-time":"2013-03-21T00:00:00Z","timestamp":1363824000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/2.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Vet Res"],"published-print":{"date-parts":[[2013,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Infectious ovine epididymitis results in substantial economic losses worldwide due to reproductive failure and culling of breeders. The most common causative agents of these infections are<jats:italic>Brucella ovis<\/jats:italic>,<jats:italic>Actinobacillus seminis<\/jats:italic>, and<jats:italic>Histophilus somni.<\/jats:italic>The aim of this study was to develop a multiplex PCR assay for simultaneous detection of<jats:italic>Brucella ovis<\/jats:italic>,<jats:italic>Actinobacillus seminis<\/jats:italic>, and<jats:italic>Histophilus somni<\/jats:italic>with species-specific primers applied to biological samples for molecular diagnosis of these infections.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The multiplex assay was capable of detecting<jats:italic>B. ovis<\/jats:italic>,<jats:italic>A. seminis<\/jats:italic>, and<jats:italic>H. somni<\/jats:italic>DNA simultaneously from genomic bacterial DNA samples and pool of semen samples from experimentally infected rams. The method was highly specific since it did not amplify DNA from other bacterial species that can potentially cause epididymitis in rams as well as species phylogenetically related to<jats:italic>B. ovis.<\/jats:italic>All negative control samples were negative in PCR multiplex assay. Urine can be used as an alternative to semen samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The species-specific multiplex PCR assay developed in this study can be successfully used for the detection of three of the most common bacterial causes of ovine epididymitis.<\/jats:p><\/jats:sec>","DOI":"10.1186\/1746-6148-9-51","type":"journal-article","created":{"date-parts":[[2013,3,21]],"date-time":"2013-03-21T07:14:34Z","timestamp":1363850074000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Species-specific multiplex PCR for the diagnosis of Brucella ovis, Actinobacillus seminis, and Histophilus somni infection in rams"],"prefix":"10.1186","volume":"9","author":[{"given":"Val\u00e9ria S","family":"Moustacas","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teane MA","family":"Silva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luciana F","family":"Costa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariana N","family":"Xavier","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"suffix":"Jr","given":"Cust\u00f3dio A","family":"Carvalho","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c9rica A","family":"Costa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatiane A","family":"Paix\u00e3o","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renato L","family":"Santos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2013,3,21]]},"reference":[{"key":"616_CR1","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1016\/0378-1135(82)90049-9","volume":"7","author":"GW Burgess","year":"1982","unstructured":"Burgess GW: Ovine contagious epididymitis: a review. Vet Microbiol. 1982, 7: 551-575. 10.1016\/0378-1135(82)90049-9.","journal-title":"Vet Microbiol"},{"key":"616_CR2","first-page":"160","volume":"34","author":"CA Carvalho J\u00fanior","year":"2010","unstructured":"Carvalho J\u00fanior CA, Xavier MN, Costa LF, Silveira SS, Sant\u2019Anna FM, Borges AM, Gouveia AMG, Santos RL: Agentes infecciosos que podem promover infertilidade em machos da esp\u00e9cie ovina [Infectious agents that can cause infertility in rams]. Rev Bras Reprod Anim 2010, 34:160\u2013167. In Portuguese. Abstract in English.","journal-title":"Rev Bras Reprod Anim"},{"key":"616_CR3","doi-asserted-by":"crossref","first-page":"1928","DOI":"10.2460\/ajvr.1986.47.09.1928","volume":"47","author":"LR Walker","year":"1986","unstructured":"Walker LR, Leamaster RB: Prevalence of Histophilus ovis and Actinobacillus seminis in the genital tract of sheep. Am J Vet Res. 1986, 47: 1928-1930.","journal-title":"Am J Vet Res"},{"key":"616_CR4","first-page":"157","volume":"73","author":"E D\u00edaz-Aparicio","year":"2009","unstructured":"D\u00edaz-Aparicio E, Tenorio-Guti\u00e9rrez VR, Arellano-Reynoso B, Enr\u00edquez-Verdugo I, Aguilar-Romero F: Pathogenicity of different strains of Histophilus somni in the experimental induction of ovine epididymitis. Can J Vet Res. 2009, 73: 157-160.","journal-title":"Can J Vet Res"},{"key":"616_CR5","first-page":"173","volume":"59","author":"AC Ward","year":"1995","unstructured":"Ward AC, Jaworski MD, Eddow JM, Corbeil LB: A comparative study of bovine and ovine Haemophilus somnus isolates. Can J Vet Res. 1995, 59: 173-178.","journal-title":"Can J Vet Res"},{"key":"616_CR6","first-page":"42","volume":"47","author":"RL Santos","year":"2005","unstructured":"Santos RL, Poester FP, Lage AP: Infec\u00e7\u00e3o por Brucella ovis [Brucella ovis infection]. Cad T\u00e9c Vet Zootec. 2005, 47: 42-56. (In Portuguese).","journal-title":"Cad T\u00e9c Vet Zootec"},{"key":"616_CR7","doi-asserted-by":"publisher","first-page":"65","DOI":"10.1016\/S0378-1135(02)00310-3","volume":"92","author":"L Manterola","year":"2003","unstructured":"Manterola L, Tejero-Garces A, Ficapal A, Shopayeva G, Blasco JM, Marin CM, L\u00f3pez-Go\u00f1i I: Evaluation of a PCR test for the diagnosis of Brucella ovis infection in semen samples from rams. Vet Microbiol. 2003, 92: 65-72. 10.1016\/S0378-1135(02)00310-3.","journal-title":"Vet Microbiol"},{"key":"616_CR8","doi-asserted-by":"publisher","first-page":"72","DOI":"10.1111\/j.1751-0813.2006.00098.x","volume":"85","author":"VF Saunders","year":"2007","unstructured":"Saunders VF, Reddacliff LA, Berg T, Hornitzky M: Multiplex PCR for the detection of Brucella ovis, Actinobacillus seminis and Histophilus somni in ram semen. Aust Vet J. 2007, 85: 72-77. 10.1111\/j.1751-0813.2006.00098.x.","journal-title":"Aust Vet J"},{"key":"616_CR9","doi-asserted-by":"publisher","first-page":"158","DOI":"10.1016\/j.vetmic.2010.02.037","volume":"145","author":"MN Xavier","year":"2010","unstructured":"Xavier MN, Silva TMA, Costa EA, Paix\u00e3o TA, Moustacas VS, Carvalho CA, Sant\u2019Anna FM, Robles CA, Gouveia AMG, Lage AP, Tsolis R, Tsolis R, Santos RL: Development and evaluation of a species-specific PCR assay for detection of Brucella ovis infection in rams. Vet Microbiol. 2010, 145: 158-164. 10.1016\/j.vetmic.2010.02.037.","journal-title":"Vet Microbiol"},{"key":"616_CR10","doi-asserted-by":"publisher","first-page":"941","DOI":"10.1111\/j.1365-2672.1998.tb05257.x","volume":"85","author":"S Appuhamy","year":"1998","unstructured":"Appuhamy S, Low JC, Parton R, Coote JG: Specific PCR primers from the 16S\u201323S rRNA spacer region for the rapid detection and identification of Actinobacillus seminis. J Appl Microbiol. 1998, 85: 941-948. 10.1111\/j.1365-2672.1998.tb05257.x.","journal-title":"J Appl Microbiol"},{"key":"616_CR11","doi-asserted-by":"publisher","first-page":"304","DOI":"10.1136\/vr.129.14.304","volume":"129","author":"PJ Heath","year":"1991","unstructured":"Heath PJ, Davies IH, Morgan JH, Aitken IA: Isolation of Actinobacillus seminis from rams in the United Kingdom. Vet Rec. 1991, 129: 304-307. 10.1136\/vr.129.14.304.","journal-title":"Vet Rec"},{"key":"616_CR12","doi-asserted-by":"crossref","first-page":"983","DOI":"10.2460\/javma.1987.190.08.983","volume":"190","author":"TE Carpenter","year":"1987","unstructured":"Carpenter TE, Berry SL, Glenn JS: Economics of Brucella ovis control in sheep: computerized decision-tree analysis. J Am Vet Med Assoc. 1987, 190: 983-987.","journal-title":"J Am Vet Med Assoc"},{"key":"616_CR13","doi-asserted-by":"publisher","first-page":"435","DOI":"10.1016\/S0378-1135(02)00228-6","volume":"90","author":"BJ Bricker","year":"2002","unstructured":"Bricker BJ: PCR as a diagnostic tool for brucellosis. Vet Microbiol. 2002, 90: 435-436. 10.1016\/S0378-1135(02)00228-6.","journal-title":"Vet Microbiol"},{"key":"616_CR14","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1136\/vr.157.5.143","volume":"157","author":"WA Al-Katib","year":"2005","unstructured":"Al-Katib WA, Dennis SM: Experimental transmission of Actinobacillus seminis infection to rams. Vet Rec. 2005, 157: 143-147.","journal-title":"Vet Rec"},{"key":"616_CR15","first-page":"27","volume":"54","author":"EL Biberstein","year":"1964","unstructured":"Biberstein EL, McGowan B, Olander H, Kennedy PC: Epididimytis in ram: Studies on pathogenesis. Cornell Vet. 1964, 54: 27-41.","journal-title":"Cornell Vet"},{"key":"616_CR16","doi-asserted-by":"publisher","first-page":"262","DOI":"10.1111\/j.1751-0813.1978.tb05558.x","volume":"54","author":"BL Clark","year":"1978","unstructured":"Clark BL, Dufty JH: Isolation of Campylobacter fetus from bulls. Aust Vet J. 1978, 54: 262-263. 10.1111\/j.1751-0813.1978.tb05558.x.","journal-title":"Aust Vet J"},{"key":"616_CR17","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1016\/j.smallrumres.2011.08.004","volume":"102","author":"CA Carvalho J\u00fanior","year":"2012","unstructured":"Carvalho J\u00fanior CA, Moustacas VS, Xavier MN, Costa EA, Costa LF, Silva TMA, Paix\u00e3o TA, Borges AM, Gouveia AMG, Santos RL: Andrological, pathologic, morphometric, and ultrasonographic findings in rams experimentally infected with Brucella ovis. Small Ruminant Res. 2012, 102: 213-222. 10.1016\/j.smallrumres.2011.08.004.","journal-title":"Small Ruminant Res"},{"key":"616_CR18","doi-asserted-by":"publisher","first-page":"480","DOI":"10.1590\/S1517-83822009000300010","volume":"40","author":"M Matrone","year":"2009","unstructured":"Matrone M, Keid LB, Rocha VCM, Vejarano MP, Ikuta CY, Rodriguez CAR, Ferreira F, Dias RA, Ferreira Neto JS: Evaluation of DNA extraction protocols for Brucella abortus PCR detection in aborted fetuses or calves born from cows experimentally infected with strain 2308. Braz J Microbiol. 2009, 40: 480-489. 10.1590\/S1517-83822009000300010.","journal-title":"Braz J Microbiol"},{"key":"616_CR19","doi-asserted-by":"publisher","first-page":"1016","DOI":"10.1590\/S0102-09352011000400031","volume":"63","author":"MN Xavier","year":"2011","unstructured":"Xavier MN, Sant\u2019Anna FM, Silva TMA, Costa EA, Moustacas VS, Merlo FA, Carvalho CA, Dasso MG, Mathias LA, Gouveia AMG, Santos RL, Lage AP: A comparison of two agar gel immunodiffusion (AGID) and a complement fixation (CF) assays for serologic diagnosis of Brucella ovis infection in experimentally infected rams. Arq Bras Med Vet Zootec. 2011, 63: 1016-1021. 10.1590\/S0102-09352011000400031.","journal-title":"Arq Bras Med Vet Zootec"},{"key":"616_CR20","doi-asserted-by":"publisher","first-page":"751","DOI":"10.1590\/S0102-09352012000300029","volume":"64","author":"EA Costa","year":"2012","unstructured":"Costa EA, Sant\u2019Anna FM, Junior Carvalho CA, Moustacas VS, Silva SMMS, Paix\u00e3o TA, Santos RL: Diagnosis of Brucella ovis infection by serology and PCR in urine samples from naturally infected rams in the State of Piau\u00ed. Arq Bras Med Vet Zootec. 2012, 64: 751-754. 10.1590\/S0102-09352012000300029.","journal-title":"Arq Bras Med Vet Zootec"},{"key":"616_CR21","doi-asserted-by":"publisher","first-page":"1706","DOI":"10.1128\/IAI.01109-10","volume":"79","author":"TMA Silva","year":"2011","unstructured":"Silva TMA, Paix\u00e3o TA, Costa EA, Xavier MN, S\u00e1 JC, Moustacas VS, Hartigh AB, Carvalho Neta AV, Tsolis R, Oliveir SC, Santos R: Putative ATP-Binding Cassette Transporter is Essential for Brucella ovis pathogenesis in Mice. Infec Immun. 2011, 79: 1706-1717. 10.1128\/IAI.01109-10.","journal-title":"Infec Immun"},{"key":"616_CR22","doi-asserted-by":"publisher","first-page":"e5519","DOI":"10.1371\/journal.pone.0005519","volume":"4","author":"RM Tsolis","year":"2009","unstructured":"Tsolis RM, Seshadri R, Santos RL, Sangari FJ, Garc\u00eda Lobo JM, Ren Q, Myers G, Brinkac LM, Nelson WC, Deboy RT, Angiuoli S, Khouri H, Dimitrov G, Robinson JR, Mulligan S, De Jong MF, Walker RL, Elzer PE, Halling SL, Hassan KA, Paulsen IT: Genome degradation in Brucella ovis corresponds with narrowing of its host range and tissue tropism. PLoS One. 2009, 4: e5519-10.1371\/journal.pone.0005519.","journal-title":"PLoS One"},{"key":"616_CR23","doi-asserted-by":"publisher","first-page":"2252","DOI":"10.1590\/S0103-84782009005000167","volume":"39","author":"MN Xavier","year":"2009","unstructured":"Xavier MN, Costa EA, Paix\u00e3o TA, Santos RL: The genus Brucella and clinical manifestations of brucellosis. Cienc Rural. 2009, 39: 2252-2260. 10.1590\/S0103-84782009005000167.","journal-title":"Cienc Rural"},{"key":"616_CR24","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1590\/S0102-09352013000100009","volume":"65","author":"LF Costa","year":"2013","unstructured":"Costa LF, Nozaki CN, Lira NSC, Antunes JMAP, Xavier MN, Costa EA, Paix\u00e3o TA, Santos RL, Megid J: Species-specific nested PCR as diagnostic tool for Brucella ovis infection in rams. Arq Bras Med Vet Zootec. 2013, 65: 55-10.1590\/S0102-09352013000100009. 60","journal-title":"Arq Bras Med Vet Zootec"},{"key":"616_CR25","doi-asserted-by":"publisher","first-page":"751","DOI":"10.1016\/S1473-3099(04)01205-8","volume":"4","author":"RP Peters","year":"2004","unstructured":"Peters RP, Van Agtmael MA, Danner SA, Savelkoul PH, Vandenbroucke-Grauls CM: New developments in the diagnosis of bloodstream infections. Lancet Infect Dis. 2004, 4: 751-760. 10.1016\/S1473-3099(04)01205-8.","journal-title":"Lancet Infect Dis"},{"key":"616_CR26","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1053\/rmed.2000.0977","volume":"95","author":"GW Waterer","year":"2001","unstructured":"Waterer GW, Wunderink RG: The influence of the severity of community-acquired pneumonia on the usefulness of blood cultures. Respir Med. 2001, 95: 78-82. 10.1053\/rmed.2000.0977.","journal-title":"Respir Med"},{"key":"616_CR27","doi-asserted-by":"publisher","first-page":"1437","DOI":"10.1067\/mob.2000.106135","volume":"182","author":"DA Wing","year":"2000","unstructured":"Wing DA, Park AS, Debuque L, Millar LK: Limited clinical utility of blood and urine cultures in the treatment of acute pyelonephritis during pregnancy. Am J Obstet Gynecol. 2000, 182: 1437-1440. 10.1067\/mob.2000.106135.","journal-title":"Am J Obstet Gynecol"}],"container-title":["BMC Veterinary Research"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/link.springer.com\/content\/pdf\/10.1186\/1746-6148-9-51.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/link.springer.com\/article\/10.1186\/1746-6148-9-51\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/1746-6148-9-51.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,4,30]],"date-time":"2025-04-30T01:17:55Z","timestamp":1745975875000},"score":1,"resource":{"primary":{"URL":"https:\/\/bmcvetres.biomedcentral.com\/articles\/10.1186\/1746-6148-9-51"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,3,21]]},"references-count":27,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2013,12]]}},"alternative-id":["616"],"URL":"https:\/\/doi.org\/10.1186\/1746-6148-9-51","relation":{},"ISSN":["1746-6148"],"issn-type":[{"value":"1746-6148","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,3,21]]},"assertion":[{"value":"26 January 2013","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"18 March 2013","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 March 2013","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}}],"article-number":"51"}}