{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:44:28Z","timestamp":1760147068494,"version":"build-2065373602"},"reference-count":43,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2023,1,3]],"date-time":"2023-01-03T00:00:00Z","timestamp":1672704000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the Ministry of Education, University and Scientific Research (MIUR)","award":["PON 2014-2020"],"award-info":[{"award-number":["PON 2014-2020"]}]},{"name":"Department of Veterinary Medicine and Animal Production, University of Naples \u201cFederico II\u201d","award":["PON 2014-2020"],"award-info":[{"award-number":["PON 2014-2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>S. microti is a new species among non-aureus staphylococci (NAS) frequently found in bovine milk samples and associated with subclinical mastitis (SCM). The aim of this study was to analyze the presence of S. microti in 200 composite milk samples and 104 milking parlor surface swabs collected at a buffalo farm in Southern Italy to define its presence in milk and a milking parlor environment. The samples were inoculated onto different agar plates, and the isolates were identified by MALDI-TOF MS. The strains identified as S. microti (54\/304 samples, 17.8%) were collected, and their purified genomic DNA was subjected to PCR amplification and whole 16S rRNA gene sequencing. Furthermore, their phenotypic resistance profiles were evaluated by a disk diffusion method, and the genotypic characterization of the tetracycline resistance was performed for the tetM and tetK genes by multiplex PCR. Four and forty-seven S. microti isolates from milk samples of lactating animals with subclinical mastitis (SCM) and intramammary infection (IMI), respectively, and three isolates from milking parlor surfaces were recovered. The genomic DNA was purified from the bacterial isolates, and the amplification and sequencing of the 16S gene further supported the proteomic identification as S. microti. No clinical mastitis was detected in the herd during the study period. The antimicrobial susceptibility testing revealed a worrisome 100% resistance to tetracyclines, genotypically mediated by the tetM gene for all strains. This study highlights that S. microti may be commonly isolated from dairy buffalo milk and milking parlor equipment. Its association with SCM or IMI remains to be established.<\/jats:p>","DOI":"10.3390\/ani13010182","type":"journal-article","created":{"date-parts":[[2023,1,4]],"date-time":"2023-01-04T01:42:44Z","timestamp":1672796564000},"page":"182","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Staphylococcus microti Strains Isolated from an Italian Mediterranean Buffalo Herd"],"prefix":"10.3390","volume":"13","author":[{"given":"Monica","family":"Ambrosio","sequence":"first","affiliation":[{"name":"Department of Veterinary Medicine and Animal Production, University of Naples \u2018Federico II\u2019, Via Delpino 1, 80137 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1789-7603","authenticated-orcid":false,"given":"Francesca Paola","family":"Nocera","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine and Animal Production, University of Naples \u2018Federico II\u2019, Via Delpino 1, 80137 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9791-1411","authenticated-orcid":false,"given":"Francesca","family":"Garofalo","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale del Mezzogiorno, 80055 Portici, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4289-6784","authenticated-orcid":false,"given":"Pasquale","family":"De Luca","sequence":"additional","affiliation":[{"name":"Stazione Zoologica Anton Dohrn of Naples, Villa Comunale, 80121 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alex","family":"Grinberg","sequence":"additional","affiliation":[{"name":"School of Veterinary Science, Massey University, Palmerston North 4442, New Zealand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luisa","family":"De Martino","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine and Animal Production, University of Naples \u2018Federico II\u2019, Via Delpino 1, 80137 Naples, Italy"},{"name":"Task Force on Microbiome Studies, University of Naples \u2018Federico II\u2019, 80131 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1292\/jvms.18-0581","article-title":"The bacterial load in milk is associated with clinical severity in cases of bovine coliform mastitis","volume":"81","author":"Nagasawa","year":"2019","journal-title":"J. Vet. Med. Sci."},{"key":"ref_2","unstructured":"Middleton, J.R., Fox, L.K., and Pighetti, G. (2017). Laboratory Handbook on Bovine Mastitis (National Mastitis Council), National Mastitis Council. [3rd ed.]."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1016\/j.rvsc.2019.11.009","article-title":"Evaluation of a bovine cathelicidin ELISA for detecting mastitis in the dairy buffalo: Comparison with milk somatic cell count and bacteriological culture","volume":"128","author":"Puggioni","year":"2020","journal-title":"Res. Vet. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1016\/j.vetmic.2015.01.024","article-title":"Species-level identification of staphylococci isolated from bovine mastitis in Brazil using partial 16S rRNA sequencing","volume":"176","author":"Lange","year":"2015","journal-title":"Vet. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"998","DOI":"10.3168\/jds.S0022-0302(06)72165-8","article-title":"Relationships between Somatic Cell Count and Intramammary Infection in Buffaloes","volume":"89","author":"Moroni","year":"2006","journal-title":"J. Dairy Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"566","DOI":"10.1099\/ijs.0.011429-0","article-title":"Staphylococcus microti sp. nov., isolated from the common vole (Microtus arvalis)","volume":"60","author":"Falsen","year":"2010","journal-title":"Int. J. Syst. Evol. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"457","DOI":"10.1016\/j.syapm.2010.08.007","article-title":"Species distribution of staphylococci from small wild mammals","volume":"33","author":"Hauschild","year":"2010","journal-title":"Syst. Appl. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2042","DOI":"10.1099\/ijs.0.012443-0","article-title":"Staphylococcus rostri sp. nov., a haemolytic bacterium isolated from the noses of healthy pigs","volume":"60","author":"Riesen","year":"2010","journal-title":"Int. J. Syst. Evol. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.vetmic.2015.11.018","article-title":"Isolation of Staphylococcus microti from milk of dairy cows with mastitis","volume":"182","author":"Wanecka","year":"2016","journal-title":"Vet. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e00420-18","DOI":"10.1128\/genomeA.00420-18","article-title":"Draft genome sequence of Staphylococcus microti DSM 22147, isolated from the common vole","volume":"6","author":"Hu","year":"2018","journal-title":"Genome Announc."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"8371","DOI":"10.1038\/s41598-022-12297-z","article-title":"Peptidomic changes in the milk of water buffaloes (Bubalus bubalis) with intramammary infection by non-aureus staphylococci","volume":"12","author":"Addis","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"8685","DOI":"10.1128\/JB.01150-07","article-title":"Population genetic structure of the Staphylococcus intermedius group: Insights into agr diversification and the emergence of methicillin-resistant strains","volume":"189","author":"Bannoehr","year":"2007","journal-title":"J. Bacteriol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/S0168-6496(03)00026-6","article-title":"Molecular analysis of bacterial microbiota in the gut of the termite Reticulitermes speratus (Isoptera; Rhinotermitidae)","volume":"44","author":"Hongoh","year":"2003","journal-title":"FEMS Microbiol. Ecol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"5445","DOI":"10.1128\/AEM.00533-11","article-title":"Biodiversity and emerging biogeography of the neutrophilic iron-oxidizing Zetaproteobacteria","volume":"77","author":"McAllister","year":"2011","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_15","unstructured":"Clinical and Laboratory Standards Institute (2015). Performance Standards for Antimicrobial Disk and Diluition Susceptibility Tests for Bacteria Isolated From Animals. VET01S, Clinical and Laboratory Standards Institute."},{"key":"ref_16","unstructured":"European Committee on Antimicrobial Susceptibility Testing (2018). Breakpoint Tables for Interpretation of MICs and Zone Diameters, European Committee on Antimicrobial Susceptibility Testing. Available online: http:\/\/www.eucast.org."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1111\/j.1469-0691.2011.03570.x","article-title":"Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: An international expert proposal for interim standard definitions for acquired resistance","volume":"18","author":"Magiorakos","year":"2012","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1051\/vetres:2001120","article-title":"Use of antimicrobials in veterinary medicine and mechanisms of resistance","volume":"32","author":"Schwarz","year":"2001","journal-title":"Vet. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1571","DOI":"10.3168\/jds.2018-15136","article-title":"Molecular epidemiology and distribution of antimicrobial resistance genes of Staphylococcus species isolated from Chinese dairy cows with clinical mastitis","volume":"102","author":"Qu","year":"2019","journal-title":"J. Dairy Sci."},{"key":"ref_20","first-page":"925","article-title":"Investigation of the genetic basis of tetracycline resistance in Staphylococcus aureus from Pakistan","volume":"11","author":"Ullah","year":"2012","journal-title":"Trop. J. Pharm. Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1111\/j.1740-0929.2008.00573.x","article-title":"Evaluation of direct and indirect measures of quarter milk from crossbred buffaloes","volume":"79","author":"Dhakal","year":"2008","journal-title":"Anim. Sci. J."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1590\/s0100-736x2017000500004","article-title":"Species identification and antimicrobial susceptibility profile of bacteria causing subclinical mastitis in buffalo","volume":"37","author":"Godoy","year":"2017","journal-title":"Pesq. Vet. Bras."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1703","DOI":"10.1590\/1678-7259","article-title":"Prevalence and etiology of buffalo mastitis and milk somatic cell count in dry and rainy seasons in a buffalo herd from Anal\u00e2ndia, S\u00e3o Paulo State, Brazil","volume":"66","author":"Pizauro","year":"2014","journal-title":"Arq. Bras. Med. Vet. Zootec."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"5592","DOI":"10.3168\/jds.2016-12478","article-title":"Prevalence of non-aureus staphylococci species causing intramammary infections in Canadian dairy herds","volume":"100","author":"Condas","year":"2017","journal-title":"J. Dairy Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"4962","DOI":"10.3168\/jds.2009-2184","article-title":"Bovine subclinical mastitis caused by different types of coagulase-negative staphylococci","volume":"92","author":"Thorberg","year":"2009","journal-title":"J. Dairy Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2329","DOI":"10.3168\/jds.2010-3741","article-title":"Some coagulase-negative Staphylococcus species affect udder health more than others","volume":"94","author":"Haesebrouck","year":"2011","journal-title":"J. Dairy Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.ijvsm.2014.02.001","article-title":"Staphylococcal species associated with bovine mastitis in the North West of Iran: Emerging of coagulase-negative staphylococci","volume":"2","author":"Hosseinzadeh","year":"2014","journal-title":"Int. J. Vet. Sci. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1776","DOI":"10.3923\/pjbs.2013.1776.1780","article-title":"Prevalence and etiology of subclinical mastitis among buffaloes (Bubalus bubalus) in Namakkal, India","volume":"16","author":"Srinivasan","year":"2013","journal-title":"Pak. J. Biol. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2631","DOI":"10.1002\/vms3.942","article-title":"Antibiogram and virulence profiling reveals multidrug resistant Staphylococcus aureus as the predominant aetiology of subclinical mastitis in riverine buffaloes","volume":"8","author":"Hoque","year":"2022","journal-title":"Vet. Med. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1016\/j.rvsc.2021.04.021","article-title":"Global and countrywide prevalence of subclinical and clinical mastitis in dairy cattle and buffaloes by systematic review and meta-analysis","volume":"136","author":"Krishnamoorthy","year":"2021","journal-title":"Res. Vet. Sci."},{"key":"ref_31","first-page":"89","article-title":"Distribution of bacteria and seasonal and regional effects in a new database for mastitis pathogens in Finland","volume":"57","author":"Koivula","year":"2007","journal-title":"Acta Agric. Scand. Sect. A Anim. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1136\/vr.160.8.253","article-title":"Survey of the incidence and aetiology of mastitis on dairy farms in England and Wales","volume":"160","author":"Bradley","year":"2007","journal-title":"Vet. Rec."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1196\/annals.1373.007","article-title":"Status of mastitis as an emerging disease in improved and periurban dairy farms in India","volume":"1081","author":"Joshi","year":"2006","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1017\/S0022029921000698","article-title":"Occurrence and aetiology of subclinical mastitis in water buffalo in Bangladesh","volume":"88","author":"Singha","year":"2021","journal-title":"J. Dairy Res."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Catozzi, C., Sanchez Bonastre, A., Francino, O., Lecchi, C., De Carlo, E., Vecchio, D., Martucciello, A., Fraulo, P., Bronzo, V., and Cusc\u00f3, A. (2017). The microbiota of water buffalo milk during mastitis. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0184710"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"108594","DOI":"10.1016\/j.vetmic.2020.108594","article-title":"Non-S. aureus staphylococci (NAS) in milk samples: Infection or contamination?","volume":"242","author":"Hamel","year":"2020","journal-title":"Vet. Microbiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.micres.2010.02.003","article-title":"Microbial phylogeny and diversity: Small subunit ribosomal RNA sequence analysis and beyond","volume":"166","author":"Rajendhran","year":"2011","journal-title":"Microbiol. Res."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Yutin, N., Puigb\u00f2, P., Koonin, E.V., and Wolf, Y.I. (2012). Phylogenomics of prokaryotic ribosomal proteins. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0036972"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2137","DOI":"10.3168\/jds.2016-12020","article-title":"Identification of bovine-associated coagulase-negative staphylococci by matrix-assisted laser desorption\/ionization time-of-flight mass spectrometry using a direct transfer protocol","volume":"100","author":"Cameron","year":"2017","journal-title":"J. Dairy Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4521","DOI":"10.3168\/jds.2014-9137","article-title":"Phenotypic antimicrobial susceptibility and occurrence of selected resistance genes in gram-positive mastitis pathogens isolated from Wisconsin dairy cows","volume":"98","author":"Ruegg","year":"2015","journal-title":"J. Dairy Sci."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Mbindyo, C.M., Gitao, G.C., Plummer, P.J., Kulohoma, B.W., Mulei, C.M., and Bett, R. (2021). Antimicrobial Resistance Profiles and Genes of Staphylococci Isolated from Mastitic Cow\u2019s Milk in Kenya. Antibiotics, 10.","DOI":"10.3390\/antibiotics10070772"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e77","DOI":"10.4142\/jvs.2020.21.e77","article-title":"Analysis of virulence traits of Staphylococcus aureus isolated from bovine mastitis in semi-intensive and family dairy farms","volume":"21","year":"2020","journal-title":"J. Vet. Sci."},{"key":"ref_43","first-page":"7","article-title":"Influence of hygiene status of cows on somatic cell count and milk components during summer season","volume":"25","author":"Erdem","year":"2019","journal-title":"Large Anim. Rev."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/13\/1\/182\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T17:57:51Z","timestamp":1760119071000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/13\/1\/182"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,3]]},"references-count":43,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["ani13010182"],"URL":"https:\/\/doi.org\/10.3390\/ani13010182","relation":{},"ISSN":["2076-2615"],"issn-type":[{"type":"electronic","value":"2076-2615"}],"subject":[],"published":{"date-parts":[[2023,1,3]]}}}