{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T01:51:26Z","timestamp":1791337886745,"version":"4.3.1"},"reference-count":28,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T00:00:00Z","timestamp":1672876800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100009647","name":"Ministry of Health","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100009647","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2023,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Water buffalo (<jats:italic>Bubalus bubalis<\/jats:italic>) has a prominent position in the livestock industry worldwide but still suffers from limited knowledge on the mechanisms regulating the immune against infections, including brucellosis (BRC), one of the most significant neglected zoonotic diseases of livestock. Seventy\u2010three buffalo were recruited for the study. Thirty\u2010five were naturally infected with<jats:italic>Brucella spp<\/jats:italic>. The aims of the study were to (i) verify the cross\u2010reactivity of 16 monoclonal antibodies (mAbs) developed against human, bovine, and ovine antigens; (ii) evaluate lymphocyte subset alterations in BRC positive buffalo; (iii) evaluate the use of the canonical discriminant analysis (CDA), with flow cytometric data, to discriminate BRC positive from negative animals. A new set of eight mAbs (anti CD3e, CD16, CD18, CD45R0, CD79a; CD172a) were shown to cross\u2010react with water buffalo orthologous molecules. BRC positive animals presented a significant (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.0001) decrease in the percentage of PBMC (29.5 vs. 40.3), total, T and B lymphocytes (23.0 vs. 35.5, 19.2 vs. 28.9, 2.6 vs. 5.7, respectively). In contrast, they showed an increase in percentage of granulocytes (65.2 vs. 55.1;<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.0001) and B lymphocytes CD21<jats:sup>neg<\/jats:sup>(22.9 vs. 16.1;<jats:italic>p<\/jats:italic>\u00a0=\u00a00.0067), a higher T\/B lymphocyte ratio (10.3 vs. 6.4;<jats:italic>p<\/jats:italic>\u00a0=\u00a00.0011) and CD3<jats:sup>+<\/jats:sup>\/CD21<jats:sup>+<\/jats:sup>(14.7 vs. 8.3;<jats:italic>p<\/jats:italic>\u00a0=\u00a00.0005) ratio. The CDA, applied to 33 different flow cytometric traits, allowed the discrimination of all BRC positive from negative buffalo. Although this is a preliminary study, our results show that flow cytometry can be used in a wide range of applications in livestock diseases, including in support of uncertain BRC diagnoses.<\/jats:p>","DOI":"10.1002\/cyto.a.24710","type":"journal-article","created":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T10:55:42Z","timestamp":1672916142000},"page":"528-536","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["In\u2010depth immunophenotyping reveals significant alteration of lymphocytes in buffalo with brucellosis"],"prefix":"10.1002","volume":"103","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0650-7908","authenticated-orcid":false,"given":"Francesco","family":"Grandoni","sequence":"first","affiliation":[{"name":"CREA\u2010Consiglio per la Ricerca in Agricoltura e l'Analisi dell'Economia Agraria Centro di ricerca Zootecnia e Acquacoltura (Research Centre for Animal Production and Aquaculture) Monterotondo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Federica","family":"Signorelli","sequence":"additional","affiliation":[{"name":"CREA\u2010Consiglio per la Ricerca in Agricoltura e l'Analisi dell'Economia Agraria Centro di ricerca Zootecnia e Acquacoltura (Research Centre for Animal Production and Aquaculture) Monterotondo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandra","family":"Martucciello","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale del Mezzogiorno National Reference Centre for Hygiene and Technologies of Water Buffalo Farming and Productions Salerno Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"Napolitano","sequence":"additional","affiliation":[{"name":"CREA\u2010Consiglio per la Ricerca in Agricoltura e l'Analisi dell'Economia Agraria Centro di ricerca Zootecnia e Acquacoltura (Research Centre for Animal Production and Aquaculture) Monterotondo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Immacolata","family":"De Donato","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale del Mezzogiorno National Reference Centre for Hygiene and Technologies of Water Buffalo Farming and Productions Salerno Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Donniacuo","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale del Mezzogiorno National Reference Centre for Hygiene and Technologies of Water Buffalo Farming and Productions Salerno Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriele","family":"Di Vuolo","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale del Mezzogiorno National Reference Centre for Hygiene and Technologies of Water Buffalo Farming and Productions Salerno Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2963-3365","authenticated-orcid":false,"given":"Giovanna","family":"De Matteis","sequence":"additional","affiliation":[{"name":"CREA\u2010Consiglio per la Ricerca in Agricoltura e l'Analisi dell'Economia Agraria Centro di ricerca Zootecnia e Acquacoltura (Research Centre for Animal Production and Aquaculture) Monterotondo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7272-7776","authenticated-orcid":false,"given":"Genny","family":"Del Zotto","sequence":"additional","affiliation":[{"name":"Dipartimento Integrato dei Servizi e Laboratori U.O.C. Laboratorio Analisi, IRCCS Istituto Giannina Gaslini Genoa Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William C.","family":"Davis","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology and Pathology Washington State University Pullman Washington USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Esterina","family":"De Carlo","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale del Mezzogiorno National Reference Centre for Hygiene and Technologies of Water Buffalo Farming and Productions Salerno Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,1,5]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12889-017-5016-y"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(07)70286-4"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001929"},{"issue":"2","key":"e_1_2_8_5_1","first-page":"233","article-title":"Brucellosis in humans: etiology, diagnostics, clinical forms","volume":"20","author":"Gali\u0144ska EM","year":"2013","journal-title":"Ann Agric Environ Med"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens10111484"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijfoodmicro.2018.12.009"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2021.649252"},{"key":"e_1_2_8_9_1","unstructured":"World organisation for animal health (OIE)Terrestrial manual 2016. Chapter 3.1.4. \u2010Brucellosis(B. abortus B. melitensis B. suis)"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20435"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.dci.2017.04.013"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2020.574434"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines9040355"},{"key":"e_1_2_8_14_1","unstructured":"Commission delegated regulation (EU) 2020\/689 of 17 December 2019 supplementing regulation (EU) 2016\/429 of the European parliament and of the council as regards rules for surveillance eradication programmes and disease\u2010free status for certain listed and emerging diseases. OJ L 174 3.6.2020 p. 211\u2013340. Available online:http:\/\/data.europa.eu\/eli\/reg_del\/2020\/689\/oj(accessed on 17 January 2021)."},{"key":"e_1_2_8_15_1","unstructured":"Italian ministry of health decree no 651 of 271994.Regulation on the national plan for the eradication of Brucellosis in cattle herds. Gazzetta ufficiale della repubblica Italiana\u2010 serie generale 277."},{"key":"e_1_2_8_16_1","unstructured":"Italian ministry of health order 92012.Extraordinary veterinary police measures of tuberculosis bovine and buffalo brucellosis ovine and caprine brucellosis leucosis in the regions of Calabria Campania Apulia and Sicily. Gazzetta ufficiale della repubblica Italiana serie generale 212."},{"key":"e_1_2_8_17_1","unstructured":"\"Extraordinary plan for the control of infectious diseases of the Italian mediterranean buffalo for the Campania region\" with relative attachment for BRC\/TBC\/LEB. Delibera of G.R. No 207 of 20 May 2019."},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.21273\/HORTSCI.29.10.1115"},{"key":"e_1_2_8_19_1","volume-title":"SAS\/STAT\u00ae 9.4 user's guide","author":"SAS Institute","year":"2012"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/150.3.419"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20490"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.20527"},{"issue":"1","key":"e_1_2_8_23_1","doi-asserted-by":"crossref","DOI":"10.1002\/cyto.a.10019","article-title":"Automated analysis of morphometric parameters for accurate definition of erythrocyte cell shape","volume":"52","author":"Albertini MC","journal-title":"Cytometry Part A"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.leu.2404675"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.3390\/vetsci7030114"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1128\/CDLI.12.1.141-151.2005"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2016.05.009"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetimm.2019.03.010"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.24492"}],"container-title":["Cytometry Part A"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.a.24710","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cyto.a.24710","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.a.24710","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,12,4]],"date-time":"2023-12-04T02:38:59Z","timestamp":1701657539000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/cyto.a.24710"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,5]]},"references-count":28,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2023,6]]}},"alternative-id":["10.1002\/cyto.a.24710"],"URL":"https:\/\/doi.org\/10.1002\/cyto.a.24710","archive":["Portico"],"relation":{},"ISSN":["1552-4922","1552-4930"],"issn-type":[{"value":"1552-4922","type":"print"},{"value":"1552-4930","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,5]]},"assertion":[{"value":"2022-01-26","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-12-10","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2023-01-05","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}