Abstract
Aeromonas hydrophila is one of the most important pathogens of zoonotic importance globally, causing various diseases in humans and animals. With the increase of drug resistance and the emergence of multidrug-resistant strains, therapeutic effects of antibiotics against A. hydrophila are limited. Hence, novel agents and strategies are urgently needed to overcome these difficulties. Wavy pili (W-pili), one of the major adhesion molecules, play a crucial role in adhesion and hemagglutination in the pathogenesis of A. hydrophila and regarded as a potential target for therapy. In this study, cinnamaldehyde, a natural compound isolated from cinnamon, could inhibit the production of W-pili and its biological function at the sub-inhibitory concentration (sub-MIC). To determine the active mechanism of cinnamaldehyde, transmission electron microscopy, adhesion inhibition, hemagglutination, electrophoresis, semi-quantitative RT-PCR, biofilm assay, and fish immersion challenge were performed. The results showed that cinnamaldehyde can reduce the production of W-pili and the formation of biofilm without affecting the growth of A. hydrophila. Besides, cinnamaldehyde interfered with the synthesis of W-pili by downregulating the expression of the genes related to pili protein, thereby reducing the adhesion and hemagglutination ability of A. hydrophila and its adhesive ability to the gills and intestines of channel catfish. Therefore, cinnamaldehyde could be employed as a promising candidate for preventing and controlling A. hydrophila infection in aquaculture, especially in antibiotic-resistant strains.








Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Abdel-Latif H, Khafaga A (2020) Natural co-infection of cultured Nile tilapia Oreochromis niloticus with Aeromonas hydrophila and Gyrodactylus cichlidarum experiencing high mortality during summer. Aquac Res 51:1880–1892
Ahmed SAKS, Rudden M, Smyth TJ, Dooley JSG, Marchant R, Banat IM (2019) Natural quorum sensing inhibitors effectively downregulate gene expression of Pseudomonas aeruginosa virulence factors. Appl Microbiol Biotechnol 103:3521–3535
AL-Subol I, Youssef N (2015) Prevalence of CTX-M, TEM and SHV β-lactamases in clinical isolates of Escherichia coli and Klebsiella pneumoniae isolated from Aleppo university hospitals, Aleppo. Syria Rev Esp Cardiol 10:399–400
Atkinson HM, Trust TJ (1980) Hemagglutination properties and adherence ability of Aeromonas hydrophila. Infect Immun 27:938–946
Austin B, Austin DA (2007) Bacterial fish pathogens. Springer Netherlands 481:482
Barbieri R, Coppo E, Marchese A, Daglia M, Sobarzo-Sánchez E, Nabavi SF, Nabavi SM (2017) Phytochemicals for human disease: an update on plant-derived compounds antibacterial activity. Microbiol Res 196:44–68
Corral FD, Shotts EB Jr, Brown J (2010) Adherence, haemagglutination and cell surface characteristics of motile aeromonads virulent for fish. J Fish Dis 13:255–268
Dong J, Ding H, Liu YT, Yang Q, Xu N, Yang Y, Ai X (2017) Magnolol protects channel catfish from Aeromonas hydrophila infection via inhibiting the expression of aerolysin. Vet Microbiol 211:119–123
Dong J, Liu Y, Xu N, Yang QH, Ai XH (2018) Morin protects channel catfish from Aeromonas hydrophila infection by blocking aerolysin activity. Front Microbiol 9:2828
Dong J, Zhang LS, Liu YT, Xu N, Zhou S, Yang QH, Yang YB, Ai XH (2020) Thymol protects channel catfish from Aeromonas hydrophila infection by inhibiting aerolysin expression and biofilm formation. Microorganisms. 8:636
EI-Son MAM, Nofal MI, Abdel-Latif HMR (2021) Co-infection of Aeromonas hydrophila and Vibrio parahaemolyticus isolated from diseased farmed striped mullet (Mugil cephalus) in Manzala, Egypt – a case report. Aquaculture. 530:735738
Ferro TA, Araújo JM, Dos Santos Pinto BL, Dos Santos JS, Souza EB, Da Silva BL, Colares VL, Novais TM, Filho CM, Struve C, Calixto JB, Monteiro-Neto V, Da Silva LC, Fernandes ES (2016) Cinnamaldehyde inhibits Staphylococcus aureus virulence factors and protects against infection in a Galleria mellonella model. Front Microbiol 7:2052
Fey PD, Olson ME (2010) Current concepts in biofilm formation of Staphylococcus epidermidis. Future Microbiol 5:917–933
Furukawa S (2015) Studies on formation, control and application of biofilm formed by food related microorganisms. Biosci Biotechnol Biochem 79:1050–1056
Han AF, Li XL, Huang BX, Tsoi KH, Matinlinna JP, Chen ZF, Deng DM (2016) The effect of titanium implant surface modification on the dynamic process of initial microbial adhesion and biofilm formation. Int J Adhes Adhes 69:125–132
Hokama A, Honma Y, Nakasone N (1990) Pili of an Aeromonas hydrophila Strain as a possible colonization factor. Microbiol Immunol 34:901–915
Hong CK, Chen HC, Su WR, Lee PY (2018) Rapid progression and mortality of necrotizing fasciitis caused by Aeromonas hydrophila in a patient with type I open tibial fracture. Kaohsiung J Med Sci 34:307–309
Hossain MJ, Sun D, McGarey DJ, Wrenn S, Alexander LM, Martino ME, Xing Y, Terhune JS, Liles MR (2014) An Asian origin of virulent Aeromonas hydrophila responsible for disease epidemics in United States-farmed catfish. Mbio. 5:e00848–e00814
Kauppi AM, Nordfelth R, Uvell H, Wolf-Watz H, Elofsson M (2003) Targeting bacterial virulence: inhibitors of type III secretion in Yersinia. Chem Biol 10:241–249
Khalifa HO, Soliman AM, Ahmed AM, Shimamoto T, Nariya H, Matsumoto T, Shimamoto T (2019) High prevalence of antimicrobial resistance in gram-negative Bacteria isolated from clinical settings in Egypt: recalling for judicious use of conventional antimicrobials in developing nations. Microb Drug Resist 25:371–385
Kim YG, Lee JH, Gwon G, Kim S, Park JG, Lee J (2016) Essential oils and eugenols inhibit biofilm formation and the virulence of Escherichia coli O157:H7. Sci Rep-UK 6:36377
Lu CY, Xie J, Xi BW et al (2015) Infection kinetics of Aeromonas hydrophila in the blunt-snout bream, Megalobrama amblycephala (in Chinese). Journal of Fishery Sciences of China 5:1068–1074
Lu J, Wang ZN, Ren MR, Huang GR, Fang BC, Bu XJ, Liu YH, Guan S (2016) Antibacterial effect of gallic acid against Aeromonas hydrophila and Aeromonas sobria through damaging membrane integrity. Curr Pharm Biotechnol 17:1153–1158
Luo YH, Yang T, Dai JH (2014) Occurrence, diagnosis and treatment of Aeromonas hydrophila in channel catfish (in Chinese). Ocean and Fishery 12:66–68
Mary P, Sautour M, Chihib NE, Tierny Y, Hornez JP (2003) Tolerance and starvation induced cross-protection against different stresses in Aeromonas hydrophila. Int J Food Microbiol 87:121–130
Mooyottu S, Kollanoor-Johny A, Flock G, Bouillaut L, Upadhyay A, Sonenshein AL, Venkitanarayanan K (2014) Carvacrol and trans-cinnamaldehyde reduce Clostridium difficile toxin production and cytotoxicity in vitro. Int J Mol Sci 15:4415–4430
Muh U, Schuster M, Heim R, Singh A, Olson ER, Greenberg EP (2006) Novel Pseudomonas aeruginosa quorum-sensing inhibitors identified in an ultra-high-throughput screen. Antimicrob Agents Ch 50:3674–3679
Murakami K, Ono T, Noma Y, Minase I, Amoh T, Irie Y, Hirota K, Miyake Y (2017) Explorative gene analysis of antibiotic tolerance-related genes in adherent and biofilm cells of Pseudomonas aeruginosa. J Infect Chemother 23:271–277
Park SY, Nam HM, Park K, Park SD (2011) Aeromonas hydrophila sepsis mimicking Vibrio vulnificus infection. Ann Dermatol 23:S25–S29
Pridgeon JW, Klesius PH (2011) Molecular identification and virulence of three Aeromonas hydrophila isolates cultured from infected channel catfish during a disease outbreak in west Alabama (USA) in 2009. Dis Aquat Org 94:249–253
Rama Devi K, Srinivasan R, Kannappan A, Santhakumari S, Bhuvaneswari M, Rajasekar P, Prabhu NM, Veera Ravi A (2016) In vitro and in vivo efficacy of rosmarinic acid on quorum sensing mediated biofilm formation and virulence factor production in Aeromonas hydrophila. Biofouling. 32:1171–1183
Rasmussen-Ivey CR, Figueras MJ, McGarey D, Liles MR (2016) Virulence factors of Aeromonas hydrophila: in the wake of reclassification. Front Microbiol 7:1337
Seshadri R, Joseph SW, Chopra AK, Sha J, Shaw J, Graf J, Haft D, Wu M, Ren Q, Rosovitz MJ, Madupu R, Tallon L, Kim M, Jin S, Vuong H, Stine OC, Ali A, Horneman AJ, Heidelberg JF (2006) Genome sequence of Aeromonas hydrophila ATCC 7966T: jack of all trades. J Bacteriol 188:8272–8282
Shoosanglertwijit R, Kamrat N, Werawatganon D, Chatsuwan T, Chaithongrat S, Rerknimitr R (2020) Real-world data of Helicobacter pylori prevalence, eradication regimens, and antibiotic resistance in Thailand, 2013–2018. JGH Open 4:49–53
Shotts EB, Rimler R (1973) Medium for the isolation of Aeromonas hydrophila. Appl Microbiol 26:550–553
Song XH, Zhao J, Bo YX, Liu ZJ, Wu K, Gong CL (2014) Aeromonas hydrophila induces intestinal inflammation in grass carp (Ctenopharyngodon idella): An experimental model. Aquaculture. 434:171–178
Song C, Cui Y, Liu B, Xie J, Ge X, Xu P, Ren M, Miao L, Zhou Q, Lin Y (2018) HSP60 and HSP90β from blunt snout bream, Megalobrama amblycephala: molecular cloning, characterization, and comparative response to intermittent thermal stress and Aeromonas hydrophila infection. Fish Shellfish Immun 74:119–132
Stratev D, Odeyemi OA (2016) Antimicrobial resistance of Aeromonas hydrophila isolated from different food sources: a mini-review. J Infect Public Heal 9:535–544
Sule AA, Tai DYH, Boey R, Tay JC (2008) Case reports: Aeromonas hydrophila severe gastroenteritis in diabetic, elderly patients. Crit Care Shock 11:67–70
Susana M, Xavier R, Alicia A, Juan MT (2010) The O:34-antigen lipopolysaccharide as an adhesin in Aeromonas hydrophila. FEMS Microbiol Lett 139:97–101
Tomás J (2012) The main Aeromonas pathogenic factors. ISRN Microbiol 2012:256261
Ventura RJ, Muhi E, Reyes VC, Sucaldito MN, Tayag E (2015) A community-based gastroenteritis outbreak after Typhoon Haiyan, Leyte, Philippines, 2013. Western Pac Surveill Response J 6:1–6
Wang LH, Wang MS, Zeng XA, Gong DM, Huang YB (2017) An in vitro investigation of the inhibitory mechanism of β-galactosidase by cinnamaldehyde alone and in combination with carvacrol and thymol. BBA-Gen Subjects 1861:3189–3198
Welker TL, Lim C, Yildirim-Aksoy M, Klesius PH (2007) Effect of buffered and unbuffered tricaine methanesulfonate (MS-222) at different concentrations on the stress responses of channel catfish. Ictalurus punctatus rafinesque Journal of Applied Aquaculture 19:1–18
Yan YX, Sun JH, Chen HQ, Lu CP (1995) Purification and characterization of pili from Aeromonas hydrophila (in Chinese). J Nanjing Agric Univ 18:88–93
Yin LZ, Chen JH, Wang KY et al (2020) Study the antibacterial mechanism of cinnamaldehyde against drug-resistant Aeromonas hydrophila in vitro. Microb Pathog 145:104208
Zalas-Wiecek P, Gospodarek E, Piecyk K (2011) Influence of subinhibitory concentrations of cefotaxime, imipenem and ciprofloxacin on adhesion of Escherichia coli strains to polystyrene. Pol J Microbiol 60:345–349
Zhang B, Teng Z, Li X, Lu G, Deng X, Niu X, Wang J (2017) Chalcone Attenuates Staphylococcus aureus Virulence by targeting sortase A and alpha-hemolysin. Front Microbiol 8:1715
Zhu XG (2000) Purification, characterization and gene location of W pili from Aeromonas hydrophila J-1 Strain (in Chinese). Nanjing Agricultural University
Availability of data and material
All data generated or analyzed during this study are included in this published article and its supplementary information files.
Code availability
Not applicable.
Funding
This work was supported by an open project of a state key laboratory of fishery drug development (grant number 201702).
Author information
Authors and Affiliations
Contributions
Not applicable.
Corresponding author
Ethics declarations
Ethics approval
Not applicable.
Competing interests
The authors declare no competing interests.
Additional information
Handling Editor: Brian Austin
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
Ouyang, P., Chen, J., Yin, L. et al. The sub-inhibitory concentration of cinnamaldehyde resists Aeromonas hydrophila pathogenicity via inhibition of W-pili production. Aquacult Int 29, 1639–1655 (2021). https://doi.org/10.1007/s10499-021-00705-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10499-021-00705-6


