Integrating uterine microbiome and metabolome to advance the understanding of the uterine environment in dairy cows with metritis.

Dairy cows Metabolome Metritis Microbiome Multi-omics Uterine disease

Journal

Animal microbiome
ISSN: 2524-4671
Titre abrégé: Anim Microbiome
Pays: England
ID NLM: 101759457

Informations de publication

Date de publication:
27 May 2024
Historique:
received: 25 01 2024
accepted: 02 05 2024
medline: 28 5 2024
pubmed: 28 5 2024
entrez: 27 5 2024
Statut: epublish

Résumé

Metritis is a prevalent uterine disease that affects the welfare, fertility, and survival of dairy cows. The uterine microbiome from cows that develop metritis and those that remain healthy do not differ from calving until 2 days postpartum, after which there is a dysbiosis of the uterine microbiome characterized by a shift towards opportunistic pathogens such as Fusobacteriota and Bacteroidota. Whether these opportunistic pathogens proliferate and overtake the uterine commensals could be determined by the type of substrates present in the uterus. The objective of this study was to integrate uterine microbiome and metabolome data to advance the understanding of the uterine environment in dairy cows that develop metritis. Holstein cows (n = 104) had uterine fluid collected at calving and at the day of metritis diagnosis. Cows with metritis (n = 52) were paired with cows without metritis (n = 52) based on days after calving. First, the uterine microbiome and metabolome were evaluated individually, and then integrated using network analyses. The uterine microbiome did not differ at calving but differed on the day of metritis diagnosis between cows with and without metritis. The uterine metabolome differed both at calving and on the day of metritis diagnosis between cows that did and did not develop metritis. Omics integration was performed between 6 significant bacteria genera and 153 significant metabolites on the day of metritis diagnosis. Integration was not performed at calving because there were no significant differences in the uterine microbiome. A total of 3 bacteria genera (i.e. Fusobacterium, Porphyromonas, and Bacteroides) were strongly correlated with 49 metabolites on the day of metritis diagnosis. Seven of the significant metabolites at calving were among the 49 metabolites strongly correlated with opportunistic pathogenic bacteria on the day of metritis diagnosis. The main metabolites have been associated with attenuation of biofilm formation by commensal bacteria, opportunistic pathogenic bacteria overgrowth, tissue damage and inflammation, immune evasion, and immune dysregulation. The data integration presented herein helps advance the understanding of the uterine environment in dairy cows with metritis. The identified metabolites may provide a competitive advantage to the main uterine pathogens Fusobacterium, Porphyromonas and Bacteroides, and may be promising targets for future interventions aiming to reduce opportunistic pathogenic bacteria growth in the uterus.

Sections du résumé

BACKGROUND BACKGROUND
Metritis is a prevalent uterine disease that affects the welfare, fertility, and survival of dairy cows. The uterine microbiome from cows that develop metritis and those that remain healthy do not differ from calving until 2 days postpartum, after which there is a dysbiosis of the uterine microbiome characterized by a shift towards opportunistic pathogens such as Fusobacteriota and Bacteroidota. Whether these opportunistic pathogens proliferate and overtake the uterine commensals could be determined by the type of substrates present in the uterus. The objective of this study was to integrate uterine microbiome and metabolome data to advance the understanding of the uterine environment in dairy cows that develop metritis. Holstein cows (n = 104) had uterine fluid collected at calving and at the day of metritis diagnosis. Cows with metritis (n = 52) were paired with cows without metritis (n = 52) based on days after calving. First, the uterine microbiome and metabolome were evaluated individually, and then integrated using network analyses.
RESULTS RESULTS
The uterine microbiome did not differ at calving but differed on the day of metritis diagnosis between cows with and without metritis. The uterine metabolome differed both at calving and on the day of metritis diagnosis between cows that did and did not develop metritis. Omics integration was performed between 6 significant bacteria genera and 153 significant metabolites on the day of metritis diagnosis. Integration was not performed at calving because there were no significant differences in the uterine microbiome. A total of 3 bacteria genera (i.e. Fusobacterium, Porphyromonas, and Bacteroides) were strongly correlated with 49 metabolites on the day of metritis diagnosis. Seven of the significant metabolites at calving were among the 49 metabolites strongly correlated with opportunistic pathogenic bacteria on the day of metritis diagnosis. The main metabolites have been associated with attenuation of biofilm formation by commensal bacteria, opportunistic pathogenic bacteria overgrowth, tissue damage and inflammation, immune evasion, and immune dysregulation.
CONCLUSIONS CONCLUSIONS
The data integration presented herein helps advance the understanding of the uterine environment in dairy cows with metritis. The identified metabolites may provide a competitive advantage to the main uterine pathogens Fusobacterium, Porphyromonas and Bacteroides, and may be promising targets for future interventions aiming to reduce opportunistic pathogenic bacteria growth in the uterus.

Identifiants

pubmed: 38802977
doi: 10.1186/s42523-024-00314-7
pii: 10.1186/s42523-024-00314-7
doi:

Types de publication

Journal Article

Langues

eng

Pagination

30

Subventions

Organisme : U.S. Department of Agriculture
ID : Grant # 2019-67015-29836, Accession No: 1019435
Organisme : U.S. Department of Agriculture
ID : Grant # 2019-67015-29836, Accession No: 1019435
Organisme : U.S. Department of Agriculture
ID : Grant # 2019-67015-29836, Accession No: 1019435
Organisme : U.S. Department of Agriculture
ID : Grant # 2019-67015-29836, Accession No: 1019435
Organisme : U.S. Department of Agriculture
ID : Grant # 2019-67015-29836, Accession No: 1019435
Organisme : U.S. Department of Agriculture
ID : Grant # 2019-67015-29836, Accession No: 1019435
Organisme : U.S. Department of Agriculture
ID : Grant # 2019-67015-29836, Accession No: 1019435

Informations de copyright

© 2024. The Author(s).

Références

Pinedo P, Santos JEP, Chebel RC, Galvão KN, Schuenemann GM, Bicalho RC, et al. Early-lactation diseases and fertility in 2 seasons of calving across US dairy herds. J Dairy Sci. 2020;103:10560–76.
pubmed: 32896394 doi: 10.3168/jds.2019-17951
Figueiredo CC, Merenda VR, de Oliveira EB, Lima FS, Chebel RC, Galvão KN, et al. Failure of clinical cure in dairy cows treated for metritis is associated with reduced productive and reproductive performance. J Dairy Sci. 2021;104:7056–70.
pubmed: 33741169 doi: 10.3168/jds.2020-19661
Barragan AA, Piñeiro JM, Schuenemann GM, Rajala-Schultz PJ, Sanders DE, Lakritz J, et al. Assessment of daily activity patterns and biomarkers of pain, inflammation, and stress in lactating dairy cows diagnosed with clinical metritis. J Dairy Sci. 2018;101:8248–58.
pubmed: 29937269 doi: 10.3168/jds.2018-14510
Pérez-Báez J, Risco CA, Chebel RC, Gomes GC, Greco LF, Tao S, et al. Association of dry matter intake and energy balance prepartum and postpartum with health disorders postpartum: part I. Calving disorders and metritis. J Dairy Sci. 2019;102:9138–50.
pubmed: 31326177 doi: 10.3168/jds.2018-15878
Jeon SJ, Cunha F, Ma X, Martinez N, Vieira-Neto A, Daetz R et al. Uterine microbiota and immune parameters associated with fever in dairy cows with metritis. PLoS ONE. 2016;11.
Jeon SJ, Vieira-Neto A, Gobikrushanth M, Daetz R, Mingoti RD, Parize ACB, et al. Uterine microbiota progression from calving until establishment of metritis in dairy cows. Appl Environ Microbiol. 2015;81:6324–32.
pubmed: 26150453 pmcid: 4542247 doi: 10.1128/AEM.01753-15
Galvão KN, Bicalho RC, Jeon SJ. Symposium review: The uterine microbiome associated with the development of uterine disease in dairy cows. J Dairy Sci. 2019;102:11786–97.
Casaro S, Prim JG, Gonzalez TD, Bisinotto RS, Chebel RC, Marrero MG, et al. Unraveling the immune and metabolic changes associated with metritis in dairy cows. J Dairy Sci. 2023;106:9244–59.
pubmed: 37641354 doi: 10.3168/jds.2023-23289
Casaro S, Prim J, Gonzalez T, Figueiredo C, Bisinotto R, Chebel R, et al. Blood metabolomics and impacted cellular mechanisms during transition into lactation in dairy cows that develop metritis. J Dairy Sci. 2023;106:8098–8109.
doi: 10.3168/jds.2023-23433 pubmed: 37641354
Figueiredo CC, Balzano-Nogueira L, Bisinotto DZ, Ruiz AR, Duarte GA, Conesa A, et al. Differences in uterine and serum metabolome associated with metritis in dairy cows. J Dairy Sci. 2023;106:3525–36.
pubmed: 36894419 doi: 10.3168/jds.2022-22552
Tan ZL, Nagaraja TG, Chengappa’ MM. Selective enumeration of Fusobacterium necrophorum from the Bovine Rument. Appl Environ Microbiol. 1994;60:1387–9.
pubmed: 8017925 pmcid: 201489 doi: 10.1128/aem.60.4.1387-1389.1994
Lee JH, Wood TK, Lee J. Roles of indole as an interspecies and interkingdom signaling molecule. Trends Microbiol. 2015;23:707–18.
pubmed: 26439294 doi: 10.1016/j.tim.2015.08.001
Pan T, Pei Z, Fang Z, Wang H, Zhu J, Zhang H, et al. Uncovering the specificity and predictability of tryptophan metabolism in lactic acid bacteria with genomics and metabolomics. Front Cell Infect Microbiol. 2023;13:1154346.
pubmed: 36992687 pmcid: 10040830 doi: 10.3389/fcimb.2023.1154346
Hailemariam D, Zhang G, Mandal R, Wishart DS, Ametaj BN. Identification of serum metabolites associated with the risk of metritis in transition dairy cows. Can J Anim Sci. 2018;98:525–37.
doi: 10.1139/cjas-2017-0069
National Research Council. 2001. Nutrient Requirements of Dairy Cattle: Seventh Revised Edition, 2001. Washington, DC: The National Academies Press. https://doi.org/10.17226/9825 .
Ferguson JD, Galligan DT, Thomsen N. Principal descriptors of body Condition score in Holstein cows. J Dairy Sci. 1994;77:2695–703.
pubmed: 7814740 doi: 10.3168/jds.S0022-0302(94)77212-X
Vieira-Neto A, Lima FS, Santos JEP, Mingoti RD, Vasconcellos GS, Risco CA, et al. Vulvovaginal laceration as risk factor for uterine disease in postpartum dairy cows. J Dairy Sci. 2016;99:4629–37.
pubmed: 27016827 doi: 10.3168/jds.2016-10872
Caporaso JG, Lauber CL, Walters WA, Berg-Lyons D, Huntley J, Fierer N, et al. Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms. ISME J. 2012;6:1621–4.
pubmed: 22402401 pmcid: 3400413 doi: 10.1038/ismej.2012.8
Fiehn O, Wohlgemuth G, Scholz M, Kind T, Lee DY, Lu Y, et al. Quality control for plant metabolomics: reporting MSI-compliant studies. Plant J. 2008;53:691–704.
pubmed: 18269577 doi: 10.1111/j.1365-313X.2007.03387.x
Fiehn O. Metabolomics by gas chromatography-mass spectrometry: combined targeted and untargeted profiling. Curr Protoc Mol Biol. 2016;2016 April:1–32.
Singh A, Shannon CP, Gautier B, Rohart F, Vacher M, Tebbutt SJ, et al. DIABLO: an integrative approach for identifying key molecular drivers from multi-omics assays. Bioinformatics. 2019;35:3055–62.
pubmed: 30657866 pmcid: 6735831 doi: 10.1093/bioinformatics/bty1054
Gley K, Hadlich F, Trakooljul N, Haack F, Murani E, Gimsa U, et al. Multi-transcript Level Profiling revealed distinct mRNA, miRNA, and tRNA-Derived fragment bio-signatures for coping Behavior Linked haplotypes in HPA Axis and Limbic System. Front Genet. 2021;12:635794.
pubmed: 34490028 pmcid: 8417057 doi: 10.3389/fgene.2021.635794
Lê Cao KA, Boitard S, Besse P. Sparse PLS discriminant analysis: biologically relevant feature selection and graphical displays for multiclass problems. BMC Bioinformatics. 2011;12:1–17.
doi: 10.1186/1471-2105-12-253
González I, Cao KAL, Davis MJ, Déjean S. Visualising associations between paired omics data sets. BioData Min. 2012;5:1–23.
doi: 10.1186/1756-0381-5-19
Karnovsky A, Weymouth T, Hull T, Glenn Tarcea V, Scardoni G, Laudanna C, et al. Metscape 2 bioinformatics tool for the analysis and visualization of metabolomics and gene expression data. Bioinformatics. 2012;28:373–80.
pubmed: 22135418 doi: 10.1093/bioinformatics/btr661
Russell WR, Duncan SH, Scobbie L, Duncan G, Cantlay L, Calder AG, et al. Major phenylpropanoid-derived metabolites in the human gut can arise from microbial fermentation of protein. Mol Nutr Food Res. 2013;57:523–35.
pubmed: 23349065 doi: 10.1002/mnfr.201200594
Mayrand D. Identification of clinical isolates of selected species of Bacteroides: production of phenylacetic acid. Can J Microbiol. 1979;25:927–8.
pubmed: 526889 doi: 10.1139/m79-138
Musthafa KS, Sivamaruthi BS, Pandian SK, Ravi AV. Quorum sensing inhibition in Pseudomonas aeruginosa PAO1 by antagonistic compound phenylacetic acid. Curr Microbiol. 2012;65:475–80.
pubmed: 22782469 doi: 10.1007/s00284-012-0181-9
Kim Y, Cho JY, Kuk JH, Moon JH, Cho J, Il, Kim YC, et al. Identification and antimicrobial activity of Phenylacetic Acid produced by Bacillus licheniformis isolated from fermented soybean, Chungkook-Jang. Curr Microbiol. 2004;48:312–7.
pubmed: 15057459 doi: 10.1007/s00284-003-4193-3
Hicks JL, Oldham KEA, Mcgarvie J, Walker EJ. Combatting antimicrobial resistance via the cysteine biosynthesis pathway in bacterial pathogens. Biosci Rep. 2022;:20220368.
Turnbull AL, Surette MG. Cysteine biosynthesis, oxidative stress and antibiotic resistance in Salmonella typhimurium. Res Microbiol. 2010;161:643–50.
pubmed: 20600858 doi: 10.1016/j.resmic.2010.06.004
Sturgill G, Toutain CM, Komperda J, O’toole GA, Rather PN. Role of CysE in production of an Extracellular Signaling Molecule in Providencia stuartii and Escherichia coli: loss of cysE enhances Biofilm formation in Escherichia coli. J Bacteriol. 2004;186:7610–7.
pubmed: 15516574 pmcid: 524891 doi: 10.1128/JB.186.22.7610-7617.2004
He M. Pipecolic acid in microbes: biosynthetic routes and enzymes. Ind Microbiol Biotechnol. 2006;33:401–7.
doi: 10.1007/s10295-006-0078-3
Neshich IA, Kiyota E, Arruda P. Genome-wide analysis of lysine catabolism in bacteria reveals new connections with osmotic stress resistance. ISME J. 2013;7:2400–10.
pubmed: 23887172 pmcid: 3834855 doi: 10.1038/ismej.2013.123
Sicsic R, Goshen T, Dutta R, Kedem-Vaanunu N, Kaplan-Shabtai V, Pasternak Z, et al. Microbial communities and inflammatory response in the endometrium differ between normal and metritic dairy cows at 5–10 days post-partum. Vet Res. 2018;49:77.
pubmed: 30068391 pmcid: 6071394 doi: 10.1186/s13567-018-0570-6
Satoh Y, Tajima K, Munekata M, Keasling JD, Lee TS. Engineering of a Tyrosol-Producing Pathway, Utilizing Simple Sugar and the Central Metabolic Tyrosine, in Escherichia coli. J Agric Food Chem. 2012;60(4):979–984.
Tsikopoulos K, Bidossi A, Drago L, Petrenyov DR, Givissis P, Mavridis D, et al. Is Implant Coating with Tyrosol- and antibiotic-loaded Hydrogel Effective in reducing Cutibacterium (Propionibacterium) acnes Biofilm formation? A preliminary in Vitro Study. Clin Orthop Relat Res. 2019;477:1736.
pubmed: 31135555 pmcid: 6999983 doi: 10.1097/CORR.0000000000000663
Arias LS, Delbem ACB, Fernandes RA, Barbosa DB, Monteiro DR. Activity of tyrosol against single and mixed-species oral biofilms. J Appl Microbiol. 2016;120:1240–9.
pubmed: 26801208 doi: 10.1111/jam.13070
Abdel-Rhman SH, El-Mahdy AM, El-Mowafy M. Effect of Tyrosol and Farnesol on Virulence and Antibiotic Resistance of Clinical isolates of Pseudomonas aeruginosa. Biomed Res Int. 2015;2015:456463.
pubmed: 26844228
Amini A, Liu M, Ahmad Z. Understanding the link between antimicrobial properties of dietary olive phenolics and bacterial ATP synthase. Int J Biol Macromol. 2017;101:153–64.
pubmed: 28322962 pmcid: 5884633 doi: 10.1016/j.ijbiomac.2017.03.087
Srinivasan R, Santhakumari S, Poonguzhali P, Geetha M, Dyavaiah M, Xiangmin L. Bacterial biofilm inhibition: a focused review on recent therapeutic strategies for combating the Biofilm mediated infections. Front Microbiol. 2021;12:676458.
pubmed: 34054785 pmcid: 8149761 doi: 10.3389/fmicb.2021.676458
Saito Y, Sato T, Nomoto K, Tsuji H. Identification of phenol- and p-cresol-producing intestinal bacteria by using media supplemented with tyrosine and its metabolites. FEMS Microbiol Ecol. 2018;94:125.
doi: 10.1093/femsec/fiy125
Wang ZY, Yin Y, Li DN, Zhao DY, Huang JQ. Biological activities of p-Hydroxycinnamic acids in maintaining Gut Barrier Integrity and function. Foods. 2023;12.
Yasuma T, Toda M, Abdel-Hamid AM, D’alessandro-Gabazza C, Kobayashi T, Nishihama K et al. Degradation products of Complex arabinoxylans by Bacteroides intestinalis enhance the host Immune Response. Microorganisms. 2021;9.
Xia X, Zhu L, Lei Z, Song Y, Tang F, Yin Z et al. Feruloylated oligosaccharides alleviate Dextran Sulfate Sodium-Induced colitis in vivo. J Agric Food Chem. 2019;67:9522–9531.
Lan H, Zhang LY, He W, Li WY, Zeng Z, Qian B et al. Sinapic acid alleviated inflammation-induced intestinal epithelial barrier dysfunction in lipopolysaccharide- (LPS-) Treated Caco-2 cells. Mediators Inflamm. 2021;5514075.
Dong L, Qin C, Li Y, Wu Z, Liu L. Oat phenolic compounds regulate metabolic syndrome in high fat diet-fed mice via gut microbiota. Food Biosci. 2022;50:101946.
Bicalho MLS, Machado VS, Higgins CH, Lima FS, Bicalho RC. Genetic and functional analysis of the bovine uterine microbiota. Part I: Metritis versus healthy cows. J Dairy Sci. 2017;100:3850–62.
pubmed: 28259404 doi: 10.3168/jds.2016-12058
Cheng J, Zhang Y, Huang M, Chen P, Zhou X, Wang D, et al. Enhanced 5-aminovalerate production in Escherichia coli from l-lysine with ethanol and hydrogen peroxide addition. J Chem Technol Biotechnol. 2018;93:3492–501.
doi: 10.1002/jctb.5708
Lin HM, Barnett MPG, Roy NC, Joyce NI, Zhu S, Armstrong K, et al. Metabolomic analysis identifies inflammatory and noninflammatory metabolic effects of genetic modification in a mouse model of Crohn’s disease. J Proteome Res. 2010;9:1965–75.
pubmed: 20141220 doi: 10.1021/pr901130s
Chatterjee B, Mondal D, Bera S. Diaminopimelic acid and its analogues: synthesis and biological perspective. Tetrahedron. 2021;100:132403.
Plata-Salaman CR, Oomura Y, Shimizu N. Endogenous Sugar Acid Derivative Acting as a feeding suppressant. Physiol Behav. 1986;38:359–73.
pubmed: 3786517 doi: 10.1016/0031-9384(86)90107-1
Zhang H, Wu L, Xu C, Xia C, Sun L, Shu S. Plasma metabolomic profiling of dairy cows affected with ketosis using gas chromatography/mass spectrometry. BMC Vet Res. 2013;9:186.
pubmed: 24070026 pmcid: 3849279 doi: 10.1186/1746-6148-9-186
Duan Y, Lu Z, Zeng S, Dan X, Zhang J, Li Y. Effects of dietary arachidonic acid on growth, immunity and intestinal microbiota of Litopenaeus vannamei under microcystin-LR stress. Aquaculture. 2022;549:737780.
doi: 10.1016/j.aquaculture.2021.737780
Krischer SM, Eisenmann M, Mueller MJ. Transport of Arachidonic Acid across the Neutrophil plasma membrane via a protein-facilitated mechanism †. Biochemistry. 1998;37:12884–91.
pubmed: 9737867 doi: 10.1021/bi980696x
Bermúdez MA, Rubio JM, Balboa MA, Balsinde J, Bermúdez MA, Rubio JM et al. Differential Mobilization of the Phospholipid and Triacylglycerol Pools of Arachidonic Acid in Murine Macrophages. Biomolecules 2022, Vol 12, Page 1851. 2022;12:1851.
Hu C, Zhao S, Li K, Yu H. Microbial Degradation of Nicotinamide by a strain Alcaligenes sp. P156. Sci Rep. 2019;9.
Ren Z, Xu Y, Li T, Sun W, Tang Z, Wang Y et al. NAD+ and its possible role in gut microbiota: insights on the mechanisms by which gut microbes influence host metabolism. Anim Nutr. 2022;10:360–371.
Hugenholtz J. Citrate metabolism in lactic acid bacteria. FEMS Microbiol Rev. 1993;12:165–78.
doi: 10.1111/j.1574-6976.1993.tb00017.x
Rodríguez MC, Viadas C, Seoane A, Sangari FJ, López-Goñi I, García-Lobo JM. Evaluation of the effects of Erythritol on Gene expression in Brucella abortus. PLoS ONE. 2012;7:e50876.
pubmed: 23272076 pmcid: 3522698 doi: 10.1371/journal.pone.0050876
Ur-Rehman S, Mushtaq Z, Zahoor T, Jamil A, Murtaza MA, Xylitol. A review on Bioproduction, Application, Health benefits, and Related Safety issues. Crit Rev Food Sci Nutr. 2015;55:1514–28.
pubmed: 24915309 doi: 10.1080/10408398.2012.702288
Tu-sekine B, Kim SF. The inositol phosphate System—A coordinator of metabolic adaptability. Int J Mol Sci. 2022;23.
Krings E, Krumbach K, Bathe B, Kelle R, Wendisch VF, Sahm H, et al. Characterization of myo-inositol utilization by Corynebacterium glutamicum: the stimulon, identification of transporters, and influence on L-lysine formation. J Bacteriol. 2006;188:8054–61.
pubmed: 16997948 pmcid: 1698185 doi: 10.1128/JB.00935-06
Leland KM, McDonald TL, Drescher KM. Effect of creatine, creatinine, and creatine ethyl ester on TLR expression in macrophages. Int Immunopharmacol. 2011;11:1341–7.
pubmed: 21575742 pmcid: 3157573 doi: 10.1016/j.intimp.2011.04.018
Geistlinger K, Schmidt JDR, Beitz E. Human monocarboxylate transporters accept and relay protons via the bound substrate for selectivity and activity at physiological pH. PNAS Nexus. 2023;2:1–8.
doi: 10.1093/pnasnexus/pgad007
Sud M, Fahy E, Cotter D, Azam K, Vadivelu I, Burant C, et al. Metabolomics Workbench: an international repository for metabolomics data and metadata, metabolite standards, protocols, tutorials and training, and analysis tools. Nucleic Acids Res. 2016;44:D463–70.
pubmed: 26467476 doi: 10.1093/nar/gkv1042

Auteurs

S Casaro (S)

Department of Large Animal Clinical Sciences, University of Florida, Gainesville, FL, USA.

J G Prim (JG)

Department of Clinical Sciences, Auburn University, Auburn, AL, USA.

T D Gonzalez (TD)

Department of Large Animal Clinical Sciences, University of Florida, Gainesville, FL, USA.

F Cunha (F)

Department of Large Animal Clinical Sciences, University of Florida, Gainesville, FL, USA.

R S Bisinotto (RS)

Department of Large Animal Clinical Sciences, University of Florida, Gainesville, FL, USA.

R C Chebel (RC)

Department of Large Animal Clinical Sciences, University of Florida, Gainesville, FL, USA.

J E P Santos (JEP)

Department of Animal Sciences, University of Florida, Gainesville, FL, USA.
D. H. Barron Reproductive and Perinatal Biology Research Program, University of Florida, Gainesville, FL, USA.

C D Nelson (CD)

Department of Animal Sciences, University of Florida, Gainesville, FL, USA.

S J Jeon (SJ)

Department of Veterinary Biomedical Sciences, Long Island University, Brookville, NY, USA.

R C Bicalho (RC)

FERA Diagnostics and Biologicals, College Station, TX, USA.

J P Driver (JP)

Division of Animals Sciences, University of Missouri, Columbia, MO, USA.

Klibs N Galvão (KN)

Department of Large Animal Clinical Sciences, University of Florida, Gainesville, FL, USA. galvaok@ufl.edu.
D. H. Barron Reproductive and Perinatal Biology Research Program, University of Florida, Gainesville, FL, USA. galvaok@ufl.edu.

Classifications MeSH