Abstract
Actinobacillus seminis is an autochthonous gram-negative bacterium that affects reproductive organs, causing epididymitis, low fertility, and occasional abortions in ovine and goats. The virulence factors and the pathogenicity mechanisms of A. seminis have not been clearly elucidated yet. In this work, biofilm production by A. seminis in in vitro assays is described and characterized. After 48-h incubation at 37 °C in trypticase soy broth, A. seminis formed biofilms containing an extracellular matrix comprised mainly of fibrillar material. Microaerophilia or the presence of calcium diminished biofilm formation in approximately 50% and 70%, respectively, but low iron concentrations increased it 40%. Through enzymatic digestion, it was found that proteins were the main component of these biofilms. Structural observations through scanning electron microscopy indicated the presence of a high amount of fibrillar material in which bacteria were immersed. Antibodies against different bacterial surface proteins, such as anti-biofilm matrix and anti-adhesin, diminished biofilm formation in 70% and 25%, respectively; whereas furanone C-30 and LED-209, compounds described as quorum-sensing inhibitors, completely inhibited biofilm formation. In conclusion, environmental conditions can influence strongly biofilm formation in A. seminis, and this could be an advantageous strategy that allows bacteria to persist inside a host.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Al-Katib WA, Dennis SM (2009) Ovine genital actinobacillosis: a review. N Z Vet J 57:352–358
Bello-Ortí B, Deslandes V, Tremblay YDN, Labrie J, Howell KJ, Tucker AW, Maskell DJ, Aragon V, Jacques M (2014) Biofilm formation by virulent and non-virulent strains of Haemophilus parasuis. Vet Res 45:104. https://doi.org/10.1186/s13567-014-0104-9
Boukahil I, Czuprynski CJ (2015) Characterization of Mannheimia haemolytica biofilm formation in vitro. Vet Microbiol 175:114–122. https://doi.org/10.1016/j.vetmic.2014.11.012
Chiba A, Sugimoto S, Sato F, Hori S, Mizunoe Y (2015) A refined technique for extraction of extracellular matrices from bacterial biofilms and its applicability. Microb Biotechnol 8:392–403. https://doi.org/10.1111/1751-7915.12155
Claessen D, Rozen DE, Kuipers OP, Søgaard-Andersen L, van Wezel GP (2014) Bacterial solutions to multicellularity: a tale of biofilms, filaments and fruiting bodies. Nat Rev Microbiol 12:115–124. https://doi.org/10.1038/nrmicro3178
Coraça-Huber DC, Dichtl S, Steixner S, Nogler M, Weiss G (2018) Iron chelation destabilizes bacterial biofilms and potentiates the antimicrobial activity of antibiotics against coagulase-negative Staphylococci. Pathog Dis. https://doi.org/10.1093/femspd/fty052
Curtis MM, Russell R, Moreira CG, Adebesin AM, Wang C, Williams NS, Taussig R, Stewart D, Zimmern P, Lu B, Prasad RN, Zhu C, Rasko DA, Huntley JF, Falck JR, Sperandio V (2014) QseC inhibitors as an antivirulence approach for Gram-negative pathogens. mBio 5(6):e02165. https://doi.org/10.1128/mBio.02165-14
Dewenter L, Volkmann TE, Maier B (2015) Oxygen governs gonococcal microcolony stability by enhancing the interaction force between type IV pili. Integr Biol (Camb) 7:1161–1170. https://doi.org/10.1039/c5ib00018a
Di Bonaventura G, Piccolomini R, Paludi D, D’Orio V, Vergara A, Conter M, Ianieri A (2008) Influence of temperature on biofilm formation by Listeria Monocytogenes on various food-contact surfaces: Relationship with motility and cell surface hydrophobicity. J Appl Microbiol 104:1552–1561. https://doi.org/10.1111/j.1365-2672.2007.03688.x
Dragoš A, Kiesewalter H, Martin M, Hsu CY, Hartmann R, Wechsler T, Eriksen C, Brix S, Drescher K, Stanley-Wall N, Kümmerli R, Kovács ÁT (2018) Division of labor during biofilm matrix production. Curr Biol 28:1903-1913.e5. https://doi.org/10.1016/j.cub.2018.04.046
Fong JNC, Yildiz FH (2015) Biofilm matrix proteins. Microbiol. Spectrum 3(2):MB-0004-2014. https://doi.org/10.1128/microbiolspec.MB-0004-2014
Fredheim EG, Klingenberg C, Rohde H, Frankenberger S, Gaustad P, Flaegstad T, Sollid JE (2009) Biofilm formation by Staphylococcus haemolyticus. J Clin Microbiol 47:1172–1180. https://doi.org/10.1128/JCM.01891-08
García-Contreras R, Peréz-Eretza B, Jasso-Chávez R, Lira-Silva E, Roldán-Sánchez JA, González-Valdez A, Soberón-Chávez G, Coria-Jiménez R, Martínez-Vázquez M, Alcaraz LD, Maeda T, Wood TK (2015) High variability in quorum quenching and growth inhibition by furanone C-30 in Pseudomonas aeruginosa clinical isolates from cystic fibrosis patients. Pathog Dis 73(6):ftv040. https://doi.org/10.1093/femspd/ftv040
Gupta P, Subhasis Sarkar S, Bannhi Das B, Bhattacharjee S, Tribedi P (2016) Biofilm, pathogenesis and prevention—a journey to break the wall: a review. Arch Microbiol 198:1–15. https://doi.org/10.1007/s00203-015-1148-6
Healey MC, Hwang HH, Elsner YY, Johnston AV (1991) A model for demonstrating the adhesion of Actinobacillus seminis to epithelial cells. Can J Vet Res 55:121–127
Heath PJ, Davies IH, Morgan JH, Aitken IA (1991) Isolation of Actinobacillus seminis from rams in the United Kingdom. Vet Rec 129:304–307
He Z, Wang Q, Hu Y, Liang J, Jiang Y, Ma R, Tang Z, Huang Z (2012) Use of the quorum sensing inhibitor furanone C-30 to interfere with biofilm formation by Streptococcus mutans and its luxS mutant strain. Int J Antimicrob Agents 40:30–35. https://doi.org/10.1016/j.ijantimicag.2012.03.016
Inzana TJ, Swords WE, Sandal I, Siddaramappa S (2008) Biofilms and quorum sensing in Pastuerellaceae. In: Kuhnert P, Christensen H (eds) Pastuerellaceae: biology, genomics and molecular aspects. Caister Academic Press, Norfolk, pp 177–196
Jain S, Chen J (2007) Attachment and biofilm formation by various serotypes of Salmonella as influenced by cellulose production and thin aggregative fimbriae biosynthesis. J Food Prod 70:2473–2479
Jang YJ, Choi YJ, Lee SH, Jun HK, Choi BK (2013) Autoinducer 2 of Fusobacterium nucleatum as a target molecule to inhibit biofilm formation of periodontopathogens. Arch Oral Biol 58:17–27. https://doi.org/10.1016/j.archoralbio.2012.04.016
Jansen BJ (1980) The aetiology of ram epididymitis. Onderstepoort J Vet Res 47:101–107
Jiménez-Pacheco A, Núñez-del-Arco A, Salas-Téllez E (2008) Microvesicles of Actinobacillus seminis show proteolytic activity. J Anim Vet Adv 7:616–619
Kannan A, Karumanchi SL, Ramalingam S, Gautam P (2016) Quantitative study on the effect of calcium and magnesium palmitate on the formation of Pseudomonas aeruginosa biofilm. J Microbiol Immunol Infect 49:988–991. https://doi.org/10.1016/j.jmii.2015.06.001
Labrie J, Pelletier-Jacques G, Deslandes V, Ramjeet M, Auger E, Nash JH, Jacques M (2010) Effects of growth conditions on biofilm formation by Actinobacillus pleuropneumoniae. Vet Res 41(1):3. https://doi.org/10.1051/vetres/2009051
López-Ochoa J, Montes-García JF, Vázquez C, Sánchez-Alonso P, Pérez-Márquez VM, Blackall PJ, Vaca S, Negrete-Abascal E (2017) Gallibacterium elongation factor-Tu possesses amyloid-like protein characteristics, participates in cell adhesion, and is present in biofilms. J Microbiol 55:745–752. https://doi.org/10.1007/s12275-017-7077-0
McCrate OA, Zhou X, Reichhardt C, Cegelski L (2013) Sum of the parts: composition and architecture of the bacterial extracellular matrix. J Mol Biol 425:4286–4294. https://doi.org/10.1016/j.jmb.2013.06.022
Montes-García JF, Vaca S, Vazquez-Cruz C, Soriano-Vargas E, Aguilar-Romero F, Blackall PJ, Negrete-Abascal E (2016) Identification of a hemagglutinin from Galllibacterium anatis. Curr Microbiol 72:450–456. https://doi.org/10.1007/s00284-015-0969-5
Montes-García JF, Chincoya Martinez DA, Vaca Pacheco S, Vázquez Cruz C, Sánchez Alonso P, Xicohtencatl Cortes J, Trujillo-Ruiz H, Negrete-Abascal E (2018a) Identification of two adhesins of Actinobacillus seminis. Small Rum Res 167:100–103. https://doi.org/10.1016/j.smallrumres.2018.08.013
Montes-García JF, Vaca S, Delgado NL, Uribe-García A, Vázquez C, Sánchez Alonso P, Xicohtencatl Cortes J, Cruz Cordoba A, Negrete Abascal E (2018b) Mannheimia haemolytica OmpP2-like is an amyloid-like protein, forms filaments, takes part in cell adhesion and is part of biofilms. Antonie Van Leeuwenhoek 111:2311–2321. https://doi.org/10.1007/s10482-018-1122-9
Montes-García JF, Delgado-Tapia WA, Vazquez-Cruz C, Vaca S, Cruz-Córdova A, Negrete-Abascal E (2019) Actinobacillus seminis GroEL-homologous protein agglutinates sheep erythrocytes. Antonie Van Leeuwenhoek 112:1655–1662. https://doi.org/10.1007/s10482-019-01292-6
Moustacas VS, Silva TM, Costa LF, Carvalho Júnior CA, Santos RL, Paixäo TA (2014) Clinical and pathological changes in rams experimentally infected with Actinobacillus seminis and Histophilus somni. Sci World J 2014:241452. https://doi.org/10.1155/2014/241452
Negrete-Abascal E, Montes-Garcia F, Vaca-Pacheco S, Leyto-Gil AM, Fragoso-Garcia E, Carvente-Garcia R, Perez-Agueros S, Castelan-Sanchez HG, Garcia-Molina A, Villamar TE, Sánchez-Alonso P, Vazquez-Cruz C (2018) Genome Sequence of Actinobacillus seminis Strain ATCC 15768, a reference strain of ovine pathogens that causes infections in reproductive organs. Genome Announc 6(2):e01453–e01417. https://doi.org/10.1128/genomeA.01453-17
Nuñez-del-Arco A, Salas-Téllez E, De la Garza M, Díaz-Aparicio E, Tenorio-Gutiérrez V (2006) Identification of an inmunogenic protein of Actinobacillus seminis that is present in microvesicles. Can J Vet Res 70:43–49
Ochoa SA, Cruz-Córdova A, Rodea GE, Cázares-Domínguez V, Escalona G, Arellano-Galindo J, Hernández-Castro R, Reyes-López A, Xicohtencatl-Cortes J (2015) Phenotypic characterization of multidrug-resistant Pseudomonas aeruginosa strains isolated from pediatric patients associated to biofilm formation. Microbiol Res 172:68–78. https://doi.org/10.1016/j.micres.2014.11.005
Pereira MF, Rossi CC, Seide LE, Martins Filho S, Dolinski CM, Bazzolli DMS (2018) Antimicrobial resistance, biofilm formation and virulence reveal Actinobacillus pleuropneumoniae strains’ pathogenicity complexity. Res Vet Sci 118:498–501. https://doi.org/10.1016/j.rvsc.2018.05.003
Petruzzi B, Inzana TJ (2016) Exopolysaccharide production and biofilm formation by Histophilus somni. Curr Top Microbiol Immunol 396:149–160. https://doi.org/10.1007/82_2015_5013
Sandal I, Hong W, Swords WE, Inzana TJ (2007) Characterization and comparison of biofilm development by pathogenic and commensal isolates of Histophilus somni. J Bacteriol 189:8179–8185
Sarkisova S, Patrauchan MA, Berglund D, Nivens DE, Franklin MJ (2005) Calcium-induced virulence factors associated with the extracellular matrix of mucoid Pseudomonas aeruginosa biofilms. J Bacteriol 187:4327–4337. https://doi.org/10.1128/JB.187.13.4327-4337.2005
Sarkisova SA, Lotlikar SR, Guragain M, Kubat R, Cloud J, Franklin MJ, Patrauchan MA (2014) A Pseudomonas aeruginosa EF-hand protein, EfhP(PA4107), modulates stress responses and virulence at high calcium concentration. PLoS ONE 9(2):e98985. https://doi.org/10.1371/journal.pone.0098985(eCollection 2014)
Schaller A, Kuhnert P, de la Puente-Redondo VA, Nicolet J, Frey J (2000) Apx toxins in Pasteurellaceae species from animals. Vet Microbiol 74:365–376
Shukla SK (2013) Effect of calcium on Staphylococcus aureus biofilm architecture: a confocal laser scanning microscopic study. Colloids Surf B Biointerfaces 103:448–454. https://doi.org/10.1016/j.colsurfb.2012.11.003
Szelestey BR, Heimlich DR, Raffel FK, Justice SS, Mason KM (2013) Haemophilus responses to nutritional immunity: epigenetic and morphological contribution to biofilm architecture, invasion, persistence and disease severity. PLoS Pathog 9(10):e1003709. https://doi.org/10.1371/journal.ppat.1003709
Taga ME, Semmelhack JL, Bassler BL (2001) The LuxS-dependent autoinducer AI-2 controls the expression of an ABC transporter that functions in AI-2 uptake in Salmonella typhimurium. Mol Microbiol 42:777–793
Vaca S, Monroy E, Rojas L, Vázquez C, Sánchez P, Soriano-Vargas E, Bojesen AM, Negrete Abascal E (2011) Adherence of Gallibacterium anatis to inert surfaces. J Anim Vet Adv 10:1688–1693. https://doi.org/10.3923/javaa.2011.1688.1693
Weigel WA, Demuth DR (2016) QseBC, a two-component bacterial adrenergic receptor and global regulator of virulence in Enterobacteriaceae and Pasteurellaceae. Mol Oral Microbiol 31:379–97. https://doi.org/10.1111/omi.12138
Funding
This study was funded by grant from DGAPA-UNAM PAPIIT IN219919. FMG is a doctoral student from the Programa de Doctorado en Ciencias de la Producción y Salud Animal, Universidad Nacional Autónoma de México (UNAM) and was supported by a scholarship from CONACYT (283503).
Author information
Authors and Affiliations
Contributions
EZ and ENA designed the experiments, FMG, LR and CVC performed the experiments. EZ, ENA and CVC wrote the drafts of the article. EZ, ENA, FMG and CVC revised this last version of the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict 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
García, J.F.M., Rojas, L., Zenteno, E. et al. Characterization of Actinobacillus seminis biofilm formation. Antonie van Leeuwenhoek 113, 1371–1383 (2020). https://doi.org/10.1007/s10482-020-01447-w
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10482-020-01447-w