Synthesis of chitosan biocomposites loaded with pyrrole-2-carboxylic acid and assessment of their antifungal activity against Aspergillus niger
A wide variety of chitosan (CS) biomaterials have been loaded with different antimicrobial agents to improve the activity of CS against phytopathogenic fungi. Recently, the antimicrobial activity of 1H-pyrrole-2-carboxylic acid (PCA) has been reported as a secondary metabolite of Streptomyces griseus, which was identified as the main bioactive compound in the biological control. However, it is sensitive to light and its activity against filamentous fungi has not yet been reported. The aim of the present research work was to evaluate the biological activity of CS-PCA biocomposites for the control of Aspergillus niger. CS-PCA biocomposites were obtained through nanoprecipitation. In vitro antifungal activity was determined by viability assay, spore germination, morphometric analysis of spores and hyphae, and the analysis of cellular components by fluorescence microscopy. CS-PCA showed an average size and Z potential of 502 ± 72 nm and + 54.7 ± 15 mV, respectively. Micrographs demonstrated well-distributed biocomposites with an apparently spherical shape. A new signal at 1473 cm−1 in the FT-IR spectrum of the CS-PCA biocomposite was observed, confirming the presence of PCA in the composition of the CS-PCA nanosystem. CS-PCA biocomposites reduced the spores’ viability by up to 58%. Effects on fungi morphometry, observed as an increase in the spores’ average diameter, swelling, distortion, and an increase in the branching of hyphae, were observed. Fluorescence analysis showed oxidative stress and membrane and cell wall damage, mainly at early growth stages. The inhibitory effect against CS-resistant fungi, such as A. niger, opens a door for the control of CS-sensitive fungi.
KeywordsAntifungal Aspergillus niger Chitosan biocomposites Secondary metabolite Nanoprecipitation
The study was funded by the Mexican Council for Science and Technology (CONACyT) through the project 219786 at the University of Sonora and for the scholarship to Alma Carolina Gálvez-Iriqui. Special thanks also go to the anonymous referees whose critical analysis of the original manuscript helped to improve the work substantially.
This study was funded by CONACyT (grant number 219786).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Aloui H, Khwaldia K, Licciardello F, Mazzaglia A, Muratore G, Hamdi M, Restuccia C (2014) Efficacy of the combined application of chitosan and locust bean gum with different citrus essential oils to control postharvest spoilage caused by Aspergillus flavus in dates. Int J Food Microbiol 170:21–28. https://doi.org/10.1016/J.IJFOODMICRO.2013.10.017 CrossRefGoogle Scholar
- Chávez-Magdaleno ME, Luque-Alcaraz AG, Gutiérrez-Martínez P, Cortez-Rocha MO, Burgos-Hernández A, Lizardi-Mendoza J, Plascencia-Jatomea M (2018) Effect of chitosan-pepper tree (Schinus molle) essential oil biocomposites on the growth kinetics, viability and membrane integrity of Colletotrichum gloeosporioides. Rev Mex Ing Quim 17:29–45. https://doi.org/10.24275/uam/izt/dcbi/revmexingquim/2018v17n1/Chavez CrossRefGoogle Scholar
- Cota-Arriola O, Cortez-Rocha MO, Ezquerra-Brauer JM, Lizardi-Mendoza J, Burgos-Hernández A, Robles-Sánchez RM, Plascencia-Jatomea M (2013) Ultrastructural, morphological, and antifungal properties of micro and nanoparticles of chitosan crosslinked with sodium tripolyphosphate. J Polym Environ 21:971–980. https://doi.org/10.1007/s10924-013-0583-1 CrossRefGoogle Scholar
- Cota-Arriola O, Plascencia-Jatomea M, Lizardi-Mendoza J, Robles-Sánchez RM, Ezquerra-Brauer JM, Ruíz-García J, Vega-Acosta JR, Cortez-Rocha MO (2017) Preparation of chitosan matrices with ferulic acid: physicochemical characterization and relationship on the growth of Aspergillus parasiticus. CyTA-J Food 15(1):65–74. https://doi.org/10.1080/19476337.2016.1213317 Google Scholar
- Dalpiaz A, Leo E, Vitali F, Pavan B, Scatturin A, Bortolotti F, Manfredini S, Durini E, Forni F, Brina B, Vandelli MA (2005) Development and characterization of biodegradable nanospheres as delivery systems of anti-ischemic adenosine derivatives. Biomaterials 26:1299–1306. https://doi.org/10.1016/J.BIOMATERIALS.2004.04.033 CrossRefGoogle Scholar
- Dananjaya SHS, Erandani WKCU, Kim C-H, Nikapitiya C, Lee J, De Zoysa M (2017) Comparative study on antifungal activities of chitosan nanoparticles and chitosan silver nano composites against Fusarium oxysporum species complex. Int J Biol Macromol 105:478–488. https://doi.org/10.1016/J.IJBIOMAC.2017.07.056 CrossRefGoogle Scholar
- Degenkolb T, Vilcinskas A (2016) Metabolites from nematophagous fungi and nematicidal natural products from fungi as alternatives for biological control. Part II: metabolites from nematophagous basidiomycetes and non-nematophagous fungi. Appl Microbiol Biotechnol 100:3813–3824. https://doi.org/10.1007/s00253-015-7234-5 CrossRefGoogle Scholar
- Hanaor D, Michelazzi M, Leonelli C, Sorrell CC (2012) The effects of carboxylic acids on the aqueous dispersion and electrophoretic deposition of ZrO2. J Eur Ceram Soc 32:235–244. https://doi.org/10.1016/J.JEURCERAMSOC.2011.08.015 CrossRefGoogle Scholar
- Hernández-Téllez CN, Rodríguez-Córdova FJ, Rosas-Burgos EC, Cortez-Rocha MO, Burgos-Hernández A, Lizardi-Mendoza J, Torres-Arreola W, Martínez-Higuera A, Plascencia-Jatomea M (2017) Activity of chitosan–lysozyme nanoparticles on the growth, membrane integrity, and β-1,3-glucanase production by Aspergillus parasiticus. 3 Biotech 7:279. https://doi.org/10.1007/s13205-017-0913-4 CrossRefGoogle Scholar
- Hernández-Téllez CN, Cortez-Rocha MO, Hernández-Burgos A, Rosas-Burgos EC, Lizardi-Mendoza J, Torres-Arreola W, Burboa-Zazueta MG, Plascencia-Jatomea M (2018) Chitosan/carrageenan/lysozyme particles: synthesis, characterization and antifungal activity against Aspergillus parasiticus. Rev Mex Ing Quim 17:897–912. https://doi.org/10.24275/uam/izt/dcbi/revmexingquim/2018v17n3/Hernandez CrossRefGoogle Scholar
- Luque-Alcaraz AG, Cortez-Rocha MO, Velázquez-Contreras CA, Acosta-Silva AL, Santacruz-Ortega H del C, Burgos-Hernández A, Argüelles-Monal WM, Plascencia-Jatomea M (2016a) Enhanced antifungal effect of chitosan/pepper tree (Schinus molle) essential oil bionanocomposites on the viability of Aspergillus parasiticus spores. J Nanomater 2016:1–10. https://doi.org/10.1155/2016/6060137 CrossRefGoogle Scholar
- Martínez-Camacho AP, Cortez-Rocha MO, Graciano-Verdugo AZ, Rodríguez-Félix F, Castillo-Ortega MM, Burgos-Hernández A, Ezquerra-Brauer JM, Plascencia-Jatomea M (2013) Extruded films of blended chitosan, low density polyethylene and ethylene acrylic acid. Carbohydr Polym 91:666–674. https://doi.org/10.1016/J.CARBPOL.2012.08.076 CrossRefGoogle Scholar
- Meshulam T, Levitz SM, Christin L, Diamond RD (1995) A simplified new assay for assessment of fungal cell damage with the tetrazolium dye, (2,3)-bis-(2-methoxy-4-nitro-5-sulphenyl)-(2H)-tetrazolium-5-carboxanilide (XTT). J Infect Dis 172:1153–1156. https://doi.org/10.1093/infdis/172.4.1153 CrossRefGoogle Scholar
- Owusu-Ansah E, Yavari A, Banerjee U (2008) A protocol for in vivo detection of reactive oxygen species. Protoc Exch 10. https://doi.org/10.1038/nprot.2008.23
- Plascencia-Jatomea M, Yépiz-Gómez MS, Vélez-Haro JM (2014) Aspergillus spp. (black Mold). In: Bautista- Baños S (ed) Post harvest decay - control strategies, vol 1. Academic Press, USA, pp 267–282. https://doi.org/10.1016/B978-0-12-411552-1.00008-9 Google Scholar
- Socrates G (2001) Infrared and Raman characteristic group frequencies: tables and charts, Third ed. Wiley, ChichesterGoogle Scholar
- Varan NY (2017) Characterization of chitosan particles via attenuated total reflection Fourier transform infrared spectroscopy, conductometric titration, viscosity average molecular weight and X-ray photoelectron spectroscopy. Asian J Chem 29:825–828. https://doi.org/10.14233/ajchem.2017.20324 CrossRefGoogle Scholar
- Vishu Kumar AB, Varadaraj MC, Gowda LR, Tharanathan RN (2005) Characterization of chito-oligosaccharides prepared by chitosanolysis with the aid of papain and pronase, and their bactericidal action against Bacillus cereus and Escherichia coli. Biochem J 391:167–175. https://doi.org/10.1042/BJ20050093 CrossRefGoogle Scholar
- Zapata-Cuartas JC (2012) Obtención de una nueva serie de compuestos pirrólicos y estudio teórico de su potencial actividad antioxidante. Master Science thesis, Universidad Nacional de Colombia, Medellín, Colombia. http://bdigital.unal.edu.co/6912/