Abstract
Bacteriocin-producing bacteria with probiotic character are known as nutritional supplements mainly for livestock. Among those beneficial bacteria we also found enterococci. Because the species strains Enterococcus mundtii also can produce bacteriocins, this study was focused on fecal strains E. mundtii from horses and their bioactivity with a view to their possible future use in breeding. Rectal removal from 47 horses (40 mares and 7 stallions), the Norik breed from Muráň were sampled in eastern Slovakia during November 2019 year. Horses age ranged from five months up to 23 years. Using MALDI-TOF mass spectrometry and 16S rRNA sequences analysis, 14 strains were allotted to the species E. mundtii. Bacteriocin substances produced by the strains EMKD 38/1, EMKD 40/2, EMKD 34/2 and EMKD 41/3 showed inhibitory activity against the most susceptible (principal) indicator strain Enterococcus avium EA5 and against listeriae as well (inhibitory activity from 100 up to 1 600 AU/mL). Only strain EMKD 41/3 possess Ent P and Mundticin KS genes and showed the broadest inhibitory activity. Ent B gene possessing strain EMKD 24/1 inhibited a growth of only indicator strain EA5. Identified E. mundtii tolerate low pH 3 and oxgall/bile. They were hemolysis, gelatinase and DNase negative and mostly susceptible to clinical antibiotics which are properties requested for application potential of strain. Substance from the strain with the broadest antimicrobial spectrum showed its practical/application potential, e.g. for optimizing the host microbiota which is important regarding the maintenance of animal`s health status.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
Not applicable.
References
Alatoom AA, Cunningham SA, Ihde S, Mandrekar J, Patel R (2011) Comparison of direct colony method versus extraction method for identification of Gram-positive cocci by use of Bruker Biotyper matrix-assissted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 49: 2868-2873. ISSN: 0095-1137.
Arboleya S, Ruas-Madiedo P, Margolles A, Solis G, Salminen S, de los Reyes-Gávilan C, Gueimonde M (2011) Characterization and in vitro properties of potentially Bifidobacterium strains isolated from breast-milk. Int J Food Microbiol 149:28–33. https://doi.org/10.1016/j.ijfoodmicro.2010.10.036
Bennik MHJ, Vanloo BR, Gorris LGM, Smid EJ (1998) A novel bacteriocin with a YGNGV motif from vegetable-associated Enterococcus mundtii: full characterization and interaction with target organisms. Biochimica et biophysica acta-protein structure and molecular enzymology 1373:47–58. https://doi.org/10.1016/S0005-2736(98)00086-8
Bhardwaj A, Malik RK, Chauhan P (2008) Functional and safety aspects of enterococci in dairy foods. Ind J Microbiol 48:317–325. https://doi.org/10.1007/s12088-008-0041-2
Bigwood T, Hudson JA, Heineman JA (2015) Enterococcus mundtii as a biocontrol agent Poster paper, www.esr.cri.nz.
Campos CA, Rodríguez O, Calo-Mata P, Prado M, Barros-Velázquez J (2006) Preliminary characterization of bacteriocin from Lactococcus lactis, Enterococcus faecium and Enterococcus mundtii strains isolated from turbot (Psetta maxima). Food Res Int 39:356–364. https://doi.org/10.1016/j.foodres.2005.08.008
Clinical Guidelines for Veterinarians Treating the Performance Horse (2017)
Cooke GC, Gibb Z, Harnett JE (2021) The safety, tolerability and efficacy of probiotic bacteria for equine use. J Equine Vet Sci 99:103407. https://doi.org/10.1016/j.jevs.2021.103407
De Vuyst L, Callewaert R, Pot B (1996) Characterization of antagonistic activity of Lactobacillus amylovorus DCE471 and large-scale isolation of its bacteriocin amylovorin L471. Syst Appl Microbiol 1:9–20. https://doi.org/10.1016/S0723-2020(96)80003-8
Dougal K, de la Fuente G, Harris PA, Girdwood SE, Pinloche E, Newbold CJ(2013) Identification of a core bacterial community within the large intestine of the horse. PLoS One 8 (10), e77660. doi: https://doi.org/10.1371/journal.pone.0077660. eCollection 2013.
Feng G, Guron GKP, Churey JJ, Worobo RW (2009) Characterization of Mundticin L, a Class IIa anti-listerial bacteriocin from Enterococcus mundtii CUGF08. Appl Environ Microbiol 75:5708–5713. https://doi.org/10.1128/AEM.00752-09
Ferreira A, Insfeld E, Canal N, Morales D, Fuentefria DB, Corcao G (2007) Characterization of enterocins produced by Enterococcus mundtii isolated from humans feces. Braz Arch Biol Technol 50:249–258. https://doi.org/10.1590/S1516-89132007000200010
Freiwald A, Sauer S (2009) Phylogenetic classification and identification of bacteria by mass spectrometry. Nat Protoc 4:723–742. https://doi.org/10.1038/nprot.2009.37
Franz CHMAP, van Belkum MJ, Holzapfel WH, Abriouel H, Gálvez A (2007) Diversity of enterococcal bacteriocins and their grouping in a new classification scheme. FEMS Microbiol Rev 31:293–310. https://doi.org/10.1111/j.1574-6976.2007.0004.x
Franz CHMAP, Huch M, Abriouel H, Holzapfel W, Galvez A (2011) Enterococci as probiotics and their implications in food safety. Int J Food Microbiol 15:125–140. https://doi.org/10.1016/j.ijfoodmicro.2011.08.014
Gilmore MS, Coburn PS, Nallapareddy SR, Murray BE (2002) Enterococcal Virulence In: The Enterococci: Pathogenesis, Molecular Biology, and Antibiotic Resistance. ASM Press, Washington, DC, pp 301–354
Gilliland SE, Walker DK (1990) Factors to consider when selecting a culture of Lactobacillus acidophilus as a dietary adjunct to produce a hypocholesterolemic effect in humans. J Dairy Sci 73:905–911. https://doi.org/10.3168/jds.S0022-0302(90)78747-4
Chikindas ML, Weeks R, Drider D, Chistyako, VA, Dicks LMT (2018) Functions and emerging applications of bacteriocins. Curr Opin Biotechnol 4: 23-28. doi.org/10.1016/j.copbio.2017.07.011.
Kauter A, Epping L, Semmler T, Antao EM, Kannapin D, Stoeckle SD, Gehlen H, Lübke-Becker A, Günther S, Wieler LH, Walther B (2019) The gut microbiome of horses: current research on equine enteral microbiota and future perspectives. Anim Microbiome 1:14. https://doi.org/10.1186/s42523-019-0013-3
Kawamoto S, Shima J, Sato R (2002) Biochemical and genetic characterization of mundticin KS, an antilisterial peptide produced by Enterococcus mundtii NFRI 7393. Appl Environ Microbiol 68:3830–3840. https://doi.org/10.1128/aem.68.8.3830-3840.2002
Kubašová I, Diep DB, Ovchinnikov KV, Lauková A, Strompfová V (2020) Bacteriocin production and distribution of bacteriocin-encoding genes in enterococci from dogs. Int J Antimcrob Agents 55:105859. https://doi.org/10.1016/j.ijantimiag.2019.11.016
Lauková A, Mareková M, Javorský P (1993) Detection and antimicrobial spectrum of a bacteriocin-like substance produced by Enterococcus faecium CCM4231. Lett Appl Microbiol 16:257–260. https://doi.org/10.1111/j.1472-765X.1993.tb01413.x
Lauková A, Simonová M, Strompfová V, Štyrtiak I, Ouwehand AC, Várady M (2008) Potential of enterococci isolated from horses. Anaerobe 14:234–236. https://doi.org/10.1016/j.anaerobe.2008.04.002
Lauková A, Focková V, Pogány Simonová M (2020) Enterococcus mundtii isolated from Slovak raw goat milk and its bacteriocinogenic potential. Int J Environ Res Pub Health 17:9504 doi:https://doi.org/10.3390/ijerph17249504.
Morrison PK, Newbold CJ, Jones E, Worgan HJ, Grove-White DH, Dugdale AH, Barfoot C, Harris PA, McGargo C (2018) The Equine Gastrointestinal Microbiome: Impacts of Age and Obesity. Front Microbiol 9:3017. https://doi.org/10.3389/fmicb.2018.03017
Semedo-Lemsaddek T, Santos MA, Lopes MFS, Figueiredo Marques JJ, Barreto Crespo MT, Tenreiro R (2003) Virulence factors in food, clinical and reference Enterococci: a common trait in the genus? Syst Appl Microbiol 26:13–22. https://doi.org/10.1078/072320203322337263
Skalka B, Pillich J, Pospíšil L (1983) Further observation on Corynebacterium renale as an indicator organism in the detection of exfoliation-positive strains of Staphylococcus aureus. Zentrbl Bacteriol Hyg A256:168–174
Strompfová V, Lauková A, Pogány Simonová M, Marciňáková M (2008) Occurrence of the structural enterocin A, P, B, L50B genes in enterococci of different origin. Vet Microbiol 132:293–301. https://doi.org/10.1016/j.vetmic.2008.05.001
Theelen MJP, Roosmarijn ECL, Wagenaar J A, Sloet van Oldriutenborgh-Oosterbaan MM, Rossen JWA, Zomer AL (2021) The equine faecal microbiota of healthy horses and ponies in the Netherlands:Impact of host and environmental factors. Animals 11:1762. doi: 10.3390/ani11061762.
The EUCAST (European Committee on Antimicrobial Susceptibility testing (2016)
Todorov SD, Wachsman MB, Knoetze H, Meincken M, Dicks LM (2005) An antibacterial and antiviral peptide produced by Enterococcus mundtii ST4V isolated from soya beans. Int J Antimicrob Agents 25:508–513. https://doi.org/10.1016/j.ijantimicag.2005.02.005
Thakkar P, Modi H, Dabhi B, Prajapati J (2014) Bile tolerance, bile deconjugation and cholesterol reducing properties of Lactobacillus strains isolated from traditional fermented foods. Int J Ferm Foods 3:157-165. doi:https://doi.org/10.5958/2321-712X.2014.01316.7.
Van Zyl WF, Deane SM, Dicks LMT (2016) Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 excludes Listeria monocytogenes from the GIT, as shown by bioluminescent studies in mice. Benef Microbes 27: 227-235. doi:https://doi.org/10.3920/BM2015.0082.
Ward MP, Alinovi CA, Couetil LL, Glickman LT, Wu CC (2004) A randomized clinical trial using probiotics to prevent salmonella fecal shedding in hospitalized horses. J Equine Vet Sci 24:242–247
Zhong Z, Zhang W, Song Y, Liu W, Xu H, Xi X, Menghe B, Sun Z (2017) Comparative genomic analysis of the genus Enterococcus. Microbiol Res 196:95–105. https://doi.org/10.1016/j.micres.2016.12.009 Epub 2017 Jan 3
Acknowledgements
The authors would like to thank Mrs. Dana Melišová for her laboratory assistance. The language was checked by English speaking person.
Author`s contributions
V. Focková: Methodology, Investigation, Writing, E. Styková: Sampling, M. Pogány Simonová: Methodology, M. Maďar:Methodology, J.Kačírová: Methodology, A. Lauková:Conceptualization, Data curation, Funding acquisition, Investigation, Project administration, Validation, Writing-editing.
Funding
This study was financially supported by the project APP0253 and partially by the joint Slovak-Polish (SK-PL) project 2019-2022 by Slovak Academy of Sciences.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Ethics approval
All care and experimental procedures involving animals followed the guidelines in the Guide for the care and use of Animals approved by the Slovak Veterinary and Food Administration and the Ethical Committee of both institutions (permission code: SK U 0716).
Consent to participate
All authors participated voluntarily in the research.
Consent for publication
All authors read and approved the final manuscript.
Conflict of interest
The authors declare they don’t have conflicts of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Focková, V., Styková, E., Simonová, M.P. et al. Horses as a source of bioactive fecal strains Enterococcus mundtii. Vet Res Commun 46, 739–747 (2022). https://doi.org/10.1007/s11259-022-09893-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11259-022-09893-9