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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 Opportunities:Scholarships abroad - Research
Effective date (Start): February 01, 2019
Effective date (End): January 31, 2020
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal Investigator:Vitor Manuel Cardoso Figueiredo Balcão
Grantee:Vitor Manuel Cardoso Figueiredo Balcão
Host Investigator: Maria Adelaide de Pinho Almeida
Host 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 (27)
(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)
EZE, OSITA C.; BEREBON, DINEBARI P.; EMENCHETA, STEPHEN C.; EVURANI, SOMTOCHUKWU A.; OKORIE, CHIBUNDO N.; BALCAO, VICTOR M.; VILA, MARTA M. D. C.. Therapeutic Potential of Marine Probiotics: A Survey on the Anticancer and Antibacterial Effects of Pseudoalteromonas spp.. PHARMACEUTICALS, v. 16, n. 8, p. 35-pg., . (18/05522-9, 22/10775-9)
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.; et al. Non-invasive Transdermal Delivery of Human Insulin Using Ionic Liquids: In vitro Studies. FRONTIERS IN PHARMACOLOGY, v. 11, . (16/08884-3, 18/05522-9, 16/12234-4)
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, . (16/12234-4, 16/08884-3, 18/05522-9)
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, . (18/05522-9, 16/12234-4, 16/08884-3)
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, . (16/12234-4, 16/08884-3, 18/05522-9)
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, . (16/08884-3, 16/12234-4, 18/05522-9)
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, . (13/03181-6, 18/05522-9, 16/08884-3, 12/15651-4)
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, . (16/08884-3, 18/05522-9, 16/12234-4)
CALDANA, CRISTIANE R. G.; HANAI-YOSHIDA, VALQUIRIA M.; PAULINO, THAIS H.; BALDO, DENICEZAR A.; FREITAS, NOBEL P.; ARANHA, NORBERTO; VILA, MARTA M. D. C.; BALCAO, VICTOR M.; OLIVEIRA JR, JOSE M.. Evaluation of urban tree barks as bioindicators of environmental pollution using the X-ray fluorescence technique. Chemosphere, v. 312, p. 13-pg., . (21/03388-6, 18/05522-9)
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, . (16/16641-3, 18/05522-9, 16/08884-3, 16/12234-4)
VILA, MARTA M. D. C.; BOSCARIOL, RODRIGO; BALCAO, VICTOR M.. NANOSTRUCTURED CURCUMIN WITH CHOLINE AND GERANIC ACID IONIC LIQUID (CAGE-IL): POTENTIAL FOR INCORPORATION INTO PHARMACEUTICAL GEL FORMULATIONS. Química Nova, v. N/A, p. 7-pg., . (16/12234-4, 16/08884-3, 18/05522-9)
BALCAO, VICTOR M.; BELLINE, BIANCA G.; SILVA, ERICA C.; ALMEIDA, PABLO F. F. B.; BALDO, DENICEZAR A.; AMORIM, LARA R. P.; OLIVEIRA JUNIOR, JOSE M.; VILA, MARTA M. D. C.; DEL FIOL, FERNANDO S.. Isolation and Molecular Characterization of Two Novel Lytic Bacteriophages for the Biocontrol of Escherichia coli in Uterine Infections: In Vitro and Ex Vivo Preliminary Studies in Veterinary Medicine. PHARMACEUTICS, v. 14, n. 11, p. 40-pg., . (16/12234-4, 21/11716-3, 16/08884-3, 18/05522-9)
LILIAN VIEIRA VAZ; VICTOR MANUEL BALCÃO; JOSÉ MARTINS OLIVEIRA JR; MATTHIEU TUBINO; ANGELA JOZALA; VALQUÍRIA MIWA HANAI YOSHIDA; MARTA MARIA DUARTE CARVALHO VILA. Development and Characterization of a Hydrogel Containing Silver Sulfadiazine for Antimicrobial Topical Applications. Part II: Stability, Cytotoxicity and Silver Release Patterns. Brazilian Journal of Pharmaceutical Sciences, v. 58, . (18/05522-9, 16/08884-3, 16/12234-4)
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, . (16/12234-4, 17/13328-5, 16/08884-3, 18/05522-9)
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, . (18/05522-9)
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, . (18/05522-9)
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, . (18/05522-9)
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, . (18/05522-9)
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, . (18/05522-9)
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, . (16/12234-4, 16/08884-3, 18/05522-9)
BOSCARIOL, RODRIGO; OLIVEIRA JUNIOR, JOSE M.; BALDO, DENICEZAR A.; BALCAO, VICTOR M.; VILA, MARTA M. D. C.. Transdermal permeation of curcumin promoted by choline geranate ionic liquid: Potential for the treatment of skin diseases. SAUDI PHARMACEUTICAL JOURNAL, v. 30, n. 4, p. 16-pg., . (16/12234-4, 18/05522-9, 16/08884-3)
BOSCARIOL, RODRIGO; CAETANO, ERIKA A.; GROTTO, DENISE; ROSA-CASTRO, RAQUEL M.; OLIVEIRA JUNIOR, JOSE M.; VILA, MARTA M. D. C.; BALCAO, VICTOR M.. Transdermal Permeation Assays of Curcumin Aided by CAGE-IL: In Vivo Control of Psoriasis. PHARMACEUTICS, v. 14, n. 4, p. 21-pg., . (16/12234-4, 16/08884-3, 18/05522-9)
BALCAO, VICTOR M.; BASU, AAKASH; CIEZA, BASILIO; ROSSI, FERNANDO N.; PEREIRA, CARLA; VILA, MARTA M. D. C.; SETUBAL, JOAO C.; HA, TAEKJIP; DA SILVA, ALINE M.. Pseudomonas-tailed lytic phages: genome mechanical analysis and putative correlation with virion morphogenesis yield. FUTURE MICROBIOLOGY, v. 17, n. 13, p. 18-pg., . (16/08884-3, 18/05522-9, 16/12234-4)
BALCAO, VICTOR M.; MORELI, FERNANDA C.; SILVA, ERICA C.; BELLINE, BIANCA G.; MARTINS, LAYLA F.; ROSSI, FERNANDO P. N.; PEREIRA, CARLA; VILA, MARTA M. D. C.; DA SILVA, ALINE M.. Isolation and Molecular Characterization of a Novel Lytic Bacteriophage That Inactivates MDR Klebsiella pneumoniae Strains. PHARMACEUTICS, v. 14, n. 7, p. 21-pg., . (16/08884-3, 18/05522-9, 16/12234-4)
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, . (16/16641-3, 18/05522-9, 16/12234-4, 16/08884-3)
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.; et al. 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, . (18/05522-9, 16/12234-4, 16/08884-3)
PEREIRA, ARTHUR O.; BARROS, NICOLE M. A.; GUERRERO, BRUNA R.; EMENCHETA, STEPHEN C.; BALDO, DENICEZAR A.; OLIVEIRA JR., JOSE M.; VILA, MARTA M. D. C.; BALCAO, VICTOR M.. An Edible Biopolymeric Microcapsular Wrapping Integrating Lytic Bacteriophage Particles for Salmonella enterica: Potential for Integration into Poultry Feed. ANTIBIOTICS-BASEL, v. 12, n. 6, p. 33-pg., . (18/05522-9, 22/10775-9, 16/12234-4, 16/08884-3)

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