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PsaPhageKill - isolation, characterisation and use of lytic bacteriophages against Pseudomonas syringae PV. actinidiae to fight the cancer of kiwifruit: an efficient and ecofriendly alternative

Grant number: 18/05522-9
Support type:Scholarships abroad - Research
Effective date (Start): February 01, 2019
Effective date (End): January 31, 2020
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal researcher:Vitor Manuel Cardoso Figueiredo Balcão
Grantee:Vitor Manuel Cardoso Figueiredo Balcão
Host: Maria Adelaide de Pinho Almeida
Home Institution: Pró-Reitoria Acadêmica. Universidade de Sorocaba (UNISO). Sorocaba , SP, Brazil
Research place: Universidade de Aveiro (UA), Portugal  

Abstract

The indiscriminate use of antibiotics combined with the adaptive capacity of microorganisms allowed the emergence of microbial strains with varying and increasing levels of antimicrobial resistance (Berquó et al., 2004; Davey et al., 2006; Silva, 2008). The increase in bacterial resistance against various antimicrobial agents limits the therapeutic options available for the effective treatment against infections.In the last decade, the worldwide production of kiwi fruit has been highly affected by Pseudomonas syringae pv. actinidiae (Psa), a phytopathogenic bacterium; this has led to severe economic losses that are seriously affecting the kiwi fruit trade. The available treatments for this disease are still scarce, with the most common involving frequently spraying the orchards with copper derivatives, in particular cuprous oxide (Cu2O). However, these copper formulations should be avoided due to their high toxicity; therefore, it is essential to search for new approaches for controlling Psa.The global population claim for new safe alternatives to antibiotics that would reduce the apparition of resistant bacteria and, at the same time, would permit a sustainable local agriculture production without the use of the traditional environmental toxic products. This research project proposal aims to develop and establish a highly accurate approach to combat and prevent the occurrence of the bacterial cancer in kiwifruit plants. To achieve that, one will use specific viruses of bacteria (bacteriophages) to eliminate the causative agent of this disease and responsible for substantial economic losses worldwide, the phytopathogen Pseudomonas syringae pv. actinidiae. To the best of my knowledge, this will be the first attempt to start the development and application of a phage therapy in the management of this emergent agricultural disease. Hence, it is expected that the results gathered will introduce novelty and significant advances in this research area. Moreover, the kiwifruit industry will benefit from this green technology, which will contribute to a sustainable growth in a highly globalized marketplace.The proposed concept is not new, existing from about 3.5 billion years, having been initiated when bacteria and bacteriophages established a predator-prey balance. Bacteriophages are the most abundant organisms on our planet (are estimated at more than 1031 particles) being completely safe for all organisms including humans, except for their target bacterial hosts. The idea embodied in the research project PsaPhageKill is unconventional and a highly creative approach, since it will join the solution (phage) with the problem (bacteria) in order to eliminate the latter. One intends to establish the proof-of-concept using either a single bacteriophage and a cocktail of bacteriophages lytic against Pseudomonas syringae pv. actinidiae, originating from environmental isolates. The physico-chemical and biological characterization of the isolated bacteriophage particles will include morphological analysis via SEM/TEM, DRX, FTIR, SDS-PAGE electrophoresis, thermal analysis by TGA and DSC, lytic spectra, detection of prophages in the host, stability and survival of the bacteriophage particles under abiotic stress (UV radiation), efficiency of plating, OSGC growth curve, adsorption curve, DNA analyses.

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Scientific publications (17)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
HARADA, LILIAM K.; SILVA, ERICA C.; ROSSI, FERNANDO P. N.; CIEZA, BASILIO; OLIVEIRA, THAIS J.; PEREIRA, CARLA; TOMAZETTO, GEIZECLER; SILVA, BIANCA B.; SQUINA, FABIO M.; VILA, MARTA M. D. C.; SETUBAL, JOAO C.; HA, TAEKJIP; DA SILVA, ALINE M.; BALCAO, VICTOR M. Characterization and in vitro testing of newly isolated lytic bacteriophages for the biocontrol of Pseudomonas aeruginosa. FUTURE MICROBIOLOGY, v. 17, n. 2, p. 111-141, JAN 2022. Web of Science Citations: 0.
HERNANDES, AUDREY N.; BOSCARIOL, RODRIGO; BALCAO, VICTOR M.; VILA, MARTA M. D. C. Transdermal Permeation of Caffeine Aided by Ionic Liquids: Potential for Enhanced Treatment of Cellulitis. AAPS PHARMSCITECH, v. 22, n. 3 APR 1 2021. Web of Science Citations: 0.
BOSCARIOL, RODRIGO; CAETANO, ERIKA A.; SILVA, ERICA C.; OLIVEIRA, THAIS J.; ROSA-CASTRO, RAQUEL M.; VILA, MARTA M. D. C.; BALCAO, VICTOR M. Performance of Choline Geranate Deep Eutectic Solvent as Transdermal Permeation Enhancer: An In Vitro Skin Histological Study. PHARMACEUTICS, v. 13, n. 4 APR 2021. Web of Science Citations: 0.
HARADA, LILIAM K.; JUNIOR, WALDEMAR BONVENTI; SILVA, ERICA C.; OLIVEIRA, THAIS J.; MORELI, FERNANDA C.; JUNIOR, JOSE M. OLIVEIRA; TUBINO, MATTHIEU; VILA, MARTA M. D. C.; BALCAO, VICTOR M. Bacteriophage-Based Biosensing of Pseudomonas aeruginosa: An Integrated Approach for the Putative Real-Time Detection of Multi-Drug-Resistant Strains. BIOSENSORS-BASEL, v. 11, n. 4 APR 2021. Web of Science Citations: 1.
ROSA MITELMAO, FABIANA CRISTINA; BERGAMASCHI, CRISTIANE DE CASSIA; GERENUTTI, MARLI; HACHEL, KARIN; SILVA, MARCUS TOLENTINO; BALCAO, VICTOR M.; DUARTE CARVALHO VILA, MARTA MARIA. The effect of probiotics on functional constipation in adults Double-blind, randomized, placebo-controlled study. MEDICINE, v. 100, n. 10 MAR 12 2021. Web of Science Citations: 1.
SILVA, ERICA C.; OLIVEIRA, THAIS J.; MORELI, FERNANDA C.; HARADA, LILIAM K.; VILA, MARTA M. D. C.; BALCAO, VICTOR M. Newly isolated lytic bacteriophages for Staphylococcus intermedius, structurally and functionally stabilized in a hydroxyethylcellulose gel containing choline geranate: Potential for transdermal permeation in veterinary phage therapy. Research in Veterinary Science, v. 135, p. 42-58, MAR 2021. Web of Science Citations: 1.
PEREIRA, CARLA; COSTA, PEDRO; PINHEIRO, LARINDJA; BALCAO, VICTOR M.; ALMEIDA, ADELAIDE. Kiwifruit bacterial canker: an integrative view focused on biocontrol strategies. PLANTA, v. 253, n. 2 JAN 27 2021. Web of Science Citations: 0.
PEREIRA, CARLA; COSTA, PEDRO; DUARTE, JOAO; BALCAO, VICTOR M.; ALMEIDA, ADELAIDE. Phage therapy as a potential approach in the biocontrol of pathogenic bacteria associated with shellfish consumption. International Journal of Food Microbiology, v. 338, JAN 2 2021. Web of Science Citations: 0.
CAMPOS, WELIDA F.; SILVA, ERICA C.; OLIVEIRA, THAIS J.; OLIVEIRA, JR., JOSE M.; TUBINO, MATTHIEU; PEREIRA, CARLA; VILA, MARTA M. D. C.; BALCAO, VICTOR M. Transdermal permeation of bacteriophage particles by choline oleate: potential for treatment of soft-tissue infections. FUTURE MICROBIOLOGY, v. 15, n. 10, p. 881-896, AUG 2020. Web of Science Citations: 0.
JORGE, LUDMILLA R.; HARADA, LILIAM K.; SILVA, ERICA C.; CAMPOS, WELIDA F.; MORELLI, FERNANDA C.; SHIMAMOTO, GUSTAVO; PEREIRA, JORGE F. B.; OLIVEIRA JR, JOSE M.; TUBINO, MATTHIEU; VILA, MARTA M. D. C.; BALCAO, VICTOR M. Non-invasive Transdermal Delivery of Human Insulin Using Ionic Liquids: In vitro Studies. FRONTIERS IN PHARMACOLOGY, v. 11, APR 23 2020. Web of Science Citations: 0.
BALCAO, VICTOR M.; HARADA, LILIAM K.; JORGE, LUDMILLA R.; OLIVEIRA JR, JOSE M.; TUBINO, MATTHIEU; VILA, MARTA M. D. C. Structural and Functional Stabilization of Sericin from Bombyx mori Cocoons in a Biopolysaccharide Film: Bioorigami for Skin Regeneration. Journal of the Brazilian Chemical Society, v. 31, n. 4, p. 833-848, APR 2020. Web of Science Citations: 0.
PINHEIRO, LARINDJA A. M.; PEREIRA, CARLA; ESTHER BARREAL, M.; PABLO GALLEGO, PEDRO; BALCAO, VICTOR M.; ALMEIDA, ADELAIDE. Use of phage phi 6 to inactivate Pseudomonas syringae pv. actinidiae in kiwifruit plants: in vitro and ex vivo experiments. Applied Microbiology and Biotechnology, v. 104, n. 3, p. 1319-1330, FEB 2020. Web of Science Citations: 0.
FABIANA CRISTINA ROSA MITELMÃO; SALVADOR MANGINI FILHO; MARCO VINÍCIUS CHAUD; MARTA MARIA DUARTE CARVALHO VILA; VICTOR MANUEL CARDOSO FIGUEIREDO BALCÃO; JOSÉ MARTINS DE OLIVEIRA JUNIOR. Caracterização física de Cateteres Centrais de Inserção Periférica (CCIP). MATERIA-RIO DE JANEIRO, v. 25, n. 1, p. -, 2020. Web of Science Citations: 0.
LUDMILLA R. JORGE; LILIAM K. HARADA; ERICA C. SILVA; WELIDA F. CAMPOS; JOSÉ M. OLIVEIRA JR.; MARTA M. D. C. VILA; MATTHIEU TUBINO; VICTOR M. BALCÃO. BACTERIAL NANOCELLULOSE BIOMEMBRANE AS A SUPPORT FOR HUMAN INSULIN AIMING AT TRANSDERMAL PERMEATION. Química Nova, v. 43, n. 5, p. 572-578, Maio 2020. Web of Science Citations: 0.
PINHEIRO, LARINDJA A. M.; PEREIRA, CARLA; FRAZAO, CAROLINA; BALCAO, VICTOR M.; ALMEIDA, ADELAIDE. Efficiency of Phage phi 6 for Biocontrol of Pseudomonas syringae pv. syringae: An in Vitro Preliminary Study. MICROORGANISMS, v. 7, n. 9 SEP 2019. Web of Science Citations: 1.
LIMA, RENATA; DEL FIOL, FERNANDO SA; BALCAO, VICTOR M. Prospects for the Use of New Technologies to Combat Multidrug-Resistant Bacteria. FRONTIERS IN PHARMACOLOGY, v. 10, JUN 21 2019. Web of Science Citations: 2.
ORSI, VALERIA C.; VILA, MARTA M. D. C.; HANAI-YOSHIDA, VALQUIRIA M.; CHAUD, V, MARCO; BALCAO, VICTOR M.; OLIVEIRA JR, JOSE M. Bran of cassava starch flour and bran of cassava flour as potential tablet excipients. ARS PHARMACEUTICA, v. 60, n. 4, p. 205-211, 2019. Web of Science Citations: 0.

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