{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T19:48:00Z","timestamp":1777578480738,"version":"3.51.4"},"reference-count":56,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2025,9,30]],"date-time":"2025-09-30T00:00:00Z","timestamp":1759190400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008736","name":"FONDEF","doi-asserted-by":"publisher","award":["ID21I10176"],"award-info":[{"award-number":["ID21I10176"]}],"id":[{"id":"10.13039\/501100008736","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Bovine Mastitis Laboratory at the Universidad Austral de Chile","award":["1121101426"],"award-info":[{"award-number":["1121101426"]}]}],"content-domain":{"domain":["www.mdpi.com"],"crossmark-restriction":true},"short-container-title":["Animals"],"abstract":"<jats:p>Background: Bovine mastitis caused by Prototheca spp. is the most significant animal disease of algal origin, with an increasing number of cases reported worldwide. Currently, there is no effective treatment, so control requires the culling of infected animals. In Chile, information is limited, and a discrepancy remains in the literature regarding the Prototheca species involved in bovine mastitis. Methods: This study aimed to molecularly type and phenotypically characterize Prototheca isolates associated with bovine mastitis in Chile. Sixty-six Prototheca isolates obtained from individual bovine mastitis milk samples and bulk tank milk samples were analyzed through cytochrome b gene (cytb) sequencing, Random Amplified Polymorphic DNA\u2013Polymerase Chain Reaction (RAPD-PCR) analysis, and phenotypic evaluation (morphology, antimicrobial susceptibility, and biofilm formation). Results: Sixty-five isolates were identified as P. bovis and one as P. ciferrii, marking the first report of the latter in bovine mastitis in Chile. RAPD analysis revealed a high genetic diversity in P. bovis. All strains exhibited resistance to the antibiotics tested from the Fluoroquinolone, \u03b2-lactam, and sulfonamide groups; however, 100% of the strains showed susceptibility to aminoglycosides, with gentamicin standing out as a potential therapeutic option. Most P. bovis strains formed weak (81.5%, 53\/65) or moderate (15.4%, 10\/65) biofilms, which could favor the persistence of infection. Conclusions: These findings provide novel insights into the molecular and phenotypic characteristics of Prototheca spp. in Chile, highlighting the predominance of P. bovis, the emergence of P. ciferri, and the implications for antimicrobial management and disease control.<\/jats:p>","DOI":"10.3390\/ani15192869","type":"journal-article","created":{"date-parts":[[2025,9,30]],"date-time":"2025-09-30T14:16:29Z","timestamp":1759241789000},"page":"2869","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Molecular and Phenotypic Characterization of Prototheca Species Isolates Associated with Bovine Mastitis Cases in Chile"],"prefix":"10.3390","volume":"15","author":[{"given":"Jaime","family":"Rodriguez","sequence":"first","affiliation":[{"name":"Instituto de Bioqu\u00edmica y Microbiolog\u00eda, Facultad de Ciencias, Universidad Austral de Chile, Valdivia 5090000, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4141-013X","authenticated-orcid":false,"given":"Paulina","family":"Sep\u00falveda-Garc\u00eda","sequence":"additional","affiliation":[{"name":"Instituto de Bioqu\u00edmica y Microbiolog\u00eda, Facultad de Ciencias, Universidad Austral de Chile, Valdivia 5090000, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nivia","family":"Canales","sequence":"additional","affiliation":[{"name":"Instituto de Bioqu\u00edmica y Microbiolog\u00eda, Facultad de Ciencias, Universidad Austral de Chile, Valdivia 5090000, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mat\u00edas","family":"Goddard","sequence":"additional","affiliation":[{"name":"Instituto de Bioqu\u00edmica y Microbiolog\u00eda, Facultad de Ciencias, Universidad Austral de Chile, Valdivia 5090000, Chile"},{"name":"Escuela de Graduados, Facultad de Ciencias, Universidad Austral de Chile, Valdivia 5090000, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carlo","family":"Cornuy","sequence":"additional","affiliation":[{"name":"Lecher\u00edas del Sur Spa, Osorno 5290000, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0883-5366","authenticated-orcid":false,"given":"\u00c1lvaro G.","family":"Morales","sequence":"additional","affiliation":[{"name":"Instituto de Ciencia Animal, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Valdivia 5090000, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9626-2675","authenticated-orcid":false,"given":"Luis","family":"Collado","sequence":"additional","affiliation":[{"name":"Instituto de Bioqu\u00edmica y Microbiolog\u00eda, Facultad de Ciencias, Universidad Austral de Chile, Valdivia 5090000, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7047-690X","authenticated-orcid":false,"given":"Armin","family":"Mella","sequence":"additional","affiliation":[{"name":"Instituto de Bioqu\u00edmica y Microbiolog\u00eda, Facultad de Ciencias, Universidad Austral de Chile, Valdivia 5090000, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1699","DOI":"10.5713\/ajas.20.0156","article-title":"Bovine Mastitis: Risk Factors, Therapeutic Strategies, and Alternative Treatments\u2014A Review","volume":"33","author":"Cheng","year":"2020","journal-title":"Asian-Australas. J. Anim. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1007\/s00284-015-0958-8","article-title":"Control of Bovine Mastitis: Old and Recent Therapeutic Approaches","volume":"72","author":"Gomes","year":"2016","journal-title":"Curr. Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"454","DOI":"10.3923\/ajava.2012.454.476","article-title":"Bovine Mastitis: An Asian Perspective","volume":"7","author":"Sharma","year":"2012","journal-title":"Asian J. Anim. Vet. Adv."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Libisch, B., Picot, C., Ceballos-garzon, A., Moravkova, M., Klimesov\u00e1, M., Telkes, G., Chuang, S.T., and Pape, P.L. (2022). Prototheca Infections and Ecology from a One Health Perspective. Microorganisms, 10.","DOI":"10.3390\/microorganisms10050938"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5635","DOI":"10.3168\/jds.2011-5106","article-title":"Herd Characteristics and Cow-Level Factors Associated with Prototheca Mastitis on Dairy Farms in Ontario, Canada","volume":"95","author":"Pieper","year":"2012","journal-title":"J. Dairy Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1007\/s11046-004-1819-3","article-title":"First Large-Scale Isolation of Prototheca zopfii from Milk Produced by Dairy Herds in Italy","volume":"158","author":"Buzzini","year":"2004","journal-title":"Mycopathologia"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"619","DOI":"10.3168\/jds.2018-15495","article-title":"A Survey on the Incidence of Protothecal Mastitis in Dairy Herds in Lublin Province, Poland","volume":"102","author":"Jagielski","year":"2019","journal-title":"J. Dairy Sci."},{"key":"ref_8","first-page":"459","article-title":"Occurrence of Prototheca spp. in Cow Milk Samples","volume":"37","author":"Bozzo","year":"2014","journal-title":"New Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1023\/A:1007005729711","article-title":"Evaluation of the Susceptibility of Prototheca zopfii to Milk Pasteurization","volume":"146","author":"Melville","year":"1999","journal-title":"Mycopathologia"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"101639","DOI":"10.1016\/j.algal.2019.101639","article-title":"The Genus Prototheca (Trebouxiophyceae, Chlorophyta) Revisited_ Implications from Molecular Taxonomic Studies","volume":"43","author":"Jagielski","year":"2019","journal-title":"Algal Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e00584-18","DOI":"10.1128\/JCM.00584-18","article-title":"Cytb as a New Genetic Marker for Differentiation of Prototheca Species","volume":"56","author":"Jagielski","year":"2018","journal-title":"J. Clin. Microbiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1007\/s11046-020-00508-y","article-title":"Protothecosis in Dogs and Cats\u2014New Research Directions","volume":"186","author":"Masuda","year":"2021","journal-title":"Mycopathologia"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Huilca-Ibarra, M.P., Vasco-Julio, D., Ledesma, Y., Guerrero-Freire, S., Zurita, J., Castillejo, P., Blasco, F.B., Yanez, L., Changoluisa, D., and Echeverr\u00eda, G. (2022). High Prevalence of Prototheca bovis Infection in Dairy Cattle with Chronic Mastitis in Ecuador. Vet. Sci., 9.","DOI":"10.20944\/preprints202209.0357.v1"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Tashakkori, N., Rahmani, H.K., and Khoramian, B. (2022). Genotypic and Phenotypic Diversity of Prototheca spp. Recovered from Bovine Mastitis in Terms of Antimicrobial Resistance and Biofilm Formation Ability. BMC Vet. Res., 18.","DOI":"10.1186\/s12917-022-03546-4"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1941","DOI":"10.1128\/JCM.00323-08","article-title":"Bovine Mastitis Associated with Prototheca blaschkeae","volume":"46","author":"Marques","year":"2008","journal-title":"J. Clin. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"642","DOI":"10.1111\/lam.12326","article-title":"Simultaneous Identification by Multiplex PCR of Major Prototheca spp. Isolated from Bovine and Buffalo Intramammary Infection and Bulk Tank","volume":"59","author":"Capra","year":"2014","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"4202","DOI":"10.3168\/jds.S0022-0302(06)72465-1","article-title":"Short Communication: In Vitro Antimicrobial Susceptibility of Prototheca Wickerhamii and Prototheca Zopfii Isolated from Bovine Mastitis","volume":"89","author":"Marques","year":"2006","journal-title":"J. Dairy Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1292\/jvms.10-0242","article-title":"Experimental Infection of Bovine Mammary Gland with Prototheca zopfii Genotype 1","volume":"73","author":"Ito","year":"2011","journal-title":"J. Vet. Med. Sci."},{"key":"ref_19","first-page":"64","article-title":"Eine Durch Algen (Prototheca) Hervorgerufene Mastitis Der Kuh","volume":"65","author":"Lerche","year":"1952","journal-title":"Berl. M\u00fcnchener Tier\u00e4rztliche Wochenschr."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1111\/j.1439-0450.1996.tb00321.x","article-title":"Prototheca sp. Outbreak of Bovine Mastitis","volume":"43","author":"Costa","year":"1996","journal-title":"J. Vet. Med. B"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Tomani\u0107, D., Bo\u017ei\u0107, D.D., Kladar, N., Samard\u017eija, M., Api\u0107, J., Baljak, J., and Kova\u010devi\u0107, Z. (2024). Clinical Evidence on Expansion of Essential Oil-Based Formulation\u2019s Pharmacological Activity in Bovine Mastitis Treatment: Antifungal Potential as Added Value. Antibiotics, 13.","DOI":"10.3390\/antibiotics13070575"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Kuczy\u0144ska, M., Kot, M., Stocki, M., Zapora, E., Jagielski, T., Perli\u0144ska-Teresiak, M., and Kali\u0144ska, A. (2025). In Vitro Determination of Cytotoxic Effects of Ten Essential Oils on Prototheca bovis, Which Causes Mastitis in Dairy Cows. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26125451"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1111\/lam.12497","article-title":"Biofilm Formation by Pathogenic Prototheca Algae","volume":"61","author":"Kwiecinski","year":"2015","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"6436","DOI":"10.3168\/jds.2016-10900","article-title":"Molecular Typing and Differences in Biofilm Formation and Antibiotic Susceptibilities among Prototheca Strains Isolated in Italy and Brazil","volume":"99","author":"Morandi","year":"2016","journal-title":"J. Dairy Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1007\/s11046-015-9951-9","article-title":"Characterization of Prototheca zopfii Genotypes Isolated from Cases of Bovine Mastitis and Cow Barns in China","volume":"181","author":"Shahid","year":"2016","journal-title":"Mycopathologia"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"173","DOI":"10.4067\/S0301-732X2011000200010","article-title":"Bovine Mastitis Caused by Prototheca zopfii: First Isolation in Chile","volume":"176","author":"Zaror","year":"2011","journal-title":"Arch. Med. Vet."},{"key":"ref_27","unstructured":"V\u00e1squez, M. (2011). Identificaci\u00f3n y Caracterizaci\u00f3n del Alga Prototheca spp. Aisladas en Un Brote de Mastitis Bovina, Universidad Austral de Chile."},{"key":"ref_28","unstructured":"National Mastitis Council (2017). Laboratory Handbook on Bovine Mastitis, National Mastitis Council. [3rd ed.]."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1186\/s13007-017-0228-9","article-title":"An Optimized Method for High Quality DNA Extraction from Microalga Prototheca wickerhamii for Genome Sequencing","volume":"13","author":"Jagielski","year":"2017","journal-title":"Plant Methods"},{"key":"ref_30","unstructured":"(2025, June 06). Available online: http:\/\/www.geneious.com."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1016\/S0022-2836(05)80360-2","article-title":"Basic Local Alignment Search Tool","volume":"215","author":"Altschul","year":"1990","journal-title":"J. Mol. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3059","DOI":"10.1093\/nar\/gkf436","article-title":"MAFFT: A Novel Method for Rapid Multiple Sequence Alignment Based on Fast Fourier Transform","volume":"30","author":"Katoh","year":"2002","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Criscuolo, A., and Gribaldo, S. (2010). BMGE (Block Mapping and Gathering with Entropy): A New Software for Selection of Phylogenetic Informative Regions from Multiple Sequence Alignments. BMC Ecol. Evol., 10.","DOI":"10.1186\/1471-2148-10-210"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"23D","DOI":"10.1093\/nar\/gkh045","article-title":"GenBank: Update","volume":"32","author":"Benson","year":"2004","journal-title":"Nucleic Acids Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1214\/aos\/1176344136","article-title":"Estimatibg the Dimension of a Model","volume":"6","author":"Schwarz","year":"1978","journal-title":"Ann. Stat."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"997","DOI":"10.1093\/sysbio\/syw037","article-title":"Terrace Aware Data Structure for Phylogenomic Inference from Supermatrices","volume":"65","author":"Chernomor","year":"2016","journal-title":"Syst. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1038\/nmeth.4285","article-title":"ModelFinder: Fast Model Selection for Accurate Phylogenetic Estimates","volume":"14","author":"Kalyaanamoorthy","year":"2017","journal-title":"Nat. Methods"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1530","DOI":"10.1093\/molbev\/msaa015","article-title":"IQ-TREE 2: New Models and Efficient Methods for Phylogenetic Inference in the Genomic Era","volume":"37","author":"Minh","year":"2020","journal-title":"Mol. Biol. Evol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1093\/molbev\/msu300","article-title":"IQ-TREE: A Fast and Effective Stochastic Algorithm for Estimating Maximum-Likelihood Phylogenies","volume":"32","author":"Nguyen","year":"2015","journal-title":"Mol. Biol. Evol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1093\/molbev\/msh123","article-title":"Bayesian Phylogenetic Model Selection Using Reversible Jump Markov Chain Monte Carlo","volume":"21","author":"Huelsenbeck","year":"2004","journal-title":"Mol. Biol. Evol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1093\/sysbio\/sys029","article-title":"MrBayes 3.2: Efficient Bayesian Phylogenetic Inference and Model Choice Across a Large Model Space","volume":"61","author":"Ronquist","year":"2012","journal-title":"Syst. Biol."},{"key":"ref_42","unstructured":"Clinical and Laboratory Standards Institute (2023). Performance Standards for Antimicrobial Susceptibility Testing. CLSI Supplement M100, Clinical and Laboratory Standards Institute. [33rd ed.]."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1016\/j.vetmic.2008.04.012","article-title":"Prototheca zopfii Genotypes Isolated from Cow Barns and Bovine Mastitis in Japan","volume":"131","author":"Osumi","year":"2008","journal-title":"Vet. Microbiol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1016\/j.vetmic.2006.10.039","article-title":"Prototheca zopfii Genotype 2-The Causative Agent of Bovine Protothecal Mastitis?","volume":"120","author":"Truyen","year":"2007","journal-title":"Vet. Microbiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1093\/mmy\/myx039","article-title":"Molecular Characterization of Prototheca Strains Isolated in China Revealed the First Cases of Protothecosis Associated with Prototheca zopfii Genotype 1","volume":"56","author":"Hirose","year":"2018","journal-title":"Med. Mycol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1177\/1040638720976423","article-title":"Molecular Characterization of Prototheca in 11 Symptomatic Dogs","volume":"33","author":"Falcaro","year":"2021","journal-title":"J. Vet. Diagn. Investig."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"e70025","DOI":"10.1111\/jvim.70025","article-title":"Protothecosis in Dogs: A Narrative Review","volume":"39","author":"Jagielski","year":"2025","journal-title":"Vet. Intern. Med."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1590\/S1516-8913201402116","article-title":"Application of RAPD-PCR for Determining the Clonality of Methicillin Resistant Staphylococcus aureus Isolated from Different Hospitals","volume":"57","author":"Idil","year":"2014","journal-title":"Braz. Arch. Biol. Technol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/j.mycmed.2008.09.001","article-title":"In Vitro Antimicrobial Susceptibility of Prototheca spp. Isolated from Bovine Mastitis in a Portugal Dairy Herd","volume":"18","author":"Lopes","year":"2008","journal-title":"J. Mycol. M\u00e9dicale"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"485","DOI":"10.2478\/bvip-2013-0084","article-title":"Antimicrobial Susceptibility of Prototheca zopfii Isolated from Bovine Mastitis","volume":"57","author":"Wawron","year":"2013","journal-title":"Bull. Vet. Inst. Pulawy"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"S9","DOI":"10.1086\/319370","article-title":"Mechanisms of Action of Antimicrobials: Focus on Fluoroquinolones","volume":"32","author":"Hooper","year":"2001","journal-title":"Clin. Infect. Dis."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1001\/archinte.163.4.402","article-title":"Trimethoprim-Sulfamethoxazole Revisited","volume":"163","author":"Masters","year":"2003","journal-title":"Arch. Intern. Med."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1007\/s11046-011-9510-y","article-title":"Characterization of Prototheca zopfii Associated with Outbreak of Bovine Clinical Mastitis in Herd of Beijing, China","volume":"173","author":"Gao","year":"2012","journal-title":"Mycopathologia"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1007\/s11046-013-9628-1","article-title":"Treatment with Gentamicin on a Murine Model of Protothecal Mastitis","volume":"175","author":"Chang","year":"2013","journal-title":"Mycopathologia"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"3613","DOI":"10.3168\/jds.2014-9248","article-title":"Biofilm-Producing Ability and Efficiency of Sanitizing Agents against Prototheca zopfii Isolates from Bovine Subclinical Mastitis","volume":"98","author":"Lee","year":"2015","journal-title":"J. Dairy Sci."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1016\/j.vetmic.2010.09.034","article-title":"Genotyping of Bovine Prototheca Mastitis Isolates from Poland","volume":"149","author":"Jagielski","year":"2011","journal-title":"Vet. Microbiol."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/19\/2869\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,9,30]],"date-time":"2025-09-30T14:28:44Z","timestamp":1759242524000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/19\/2869"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,30]]},"references-count":56,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2025,10]]}},"alternative-id":["ani15192869"],"URL":"https:\/\/doi.org\/10.3390\/ani15192869","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,30]]}}}