Journal of Microbiology

Journal Articles

[Protocol] Use of Cas9 Targeting and Red Recombination for Designer Phage Engineering: Shin-Yae Choi , Danitza Xiomara Romero-Calle , Han-Gyu Cho , Hee-Won Bae , You-Hee Cho; J. Microbiol. 2024;62(1):1-10. Published online February 1, 2024; DOI: https://doi.org/10.1007/s12275-024-00107-2

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Bacteriophages (phages) are natural antibiotics and biological nanoparticles, whose application is significantly boosted by recent advances of synthetic biology tools. Designer phages are synthetic phages created by genome engineering in a way to increase the benefits or decrease the drawbacks of natural phages. Here we report the development of a straightforward genome engineering method to efficiently obtain engineered phages in a model bacterial pathogen, Pseudomonas aeruginosa. This was achieved by eliminating the wild type phages based on the Streptococcus pyogenes Cas9 (SpCas9) and facilitating the recombinant generation based on the Red recombination system of the coliphage λ (λRed). The producer (PD) cells of P. aeruginosa strain PAO1 was created by miniTn7-based chromosomal integration of the genes for SpCas9 and λRed under an inducible promoter. To validate the efficiency of the recombinant generation, we created the fluorescent phages from a temperate phage MP29. A plasmid bearing the single guide RNA (sgRNA) gene for selectively targeting the wild type gp35 gene and the editing template for tagging the Gp35 with superfolder green fluorescent protein (sfGFP) was introduced into the PD cells by electroporation. We found that the targeting efficiency was affected by the position and number of sgRNA. The fluorescent phage particles were efficiently recovered from the culture of the PD cells expressing dual sgRNA molecules. This protocol can be used to create designer phages in P. aeruginosa for both application and research purposes.

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Pilin regions that select for the small RNA phages in Pseudomonas aeruginosa type IV pilus
Hee-Won Bae, Hyeong-Jun Ki, Shin-Yae Choi, You-Hee Cho, Kristin N. Parent
Journal of Virology.2025;[Epub] CrossRef
Characteristics of bioaerosols under high-ozone periods, haze episodes, dust storms, and normal days in Xi’an, China
Yiming Yang, Liu Yang, Xiaoyan Hu, Zhenxing Shen
Particuology.2024; 90: 140. CrossRef
Airborne desert dust and aeromicrobiology over the Turkish Mediterranean coastline
Dale W. Griffin, Nilgün Kubilay, Mustafa Koçak, Mike A. Gray, Timothy C. Borden, Eugene A. Shinn
Atmospheric Environment.2007; 41(19): 4050. CrossRef

Candida krusei isolated from fruit juices ultrafiltration membranes promotes colonization of Escherichia coli O157:H7 and Salmonella enterica on stainless steel surfaces: María Clara Tarifa , Jorge Enrique Lozano , Lorena Inés Brugnoni; J. Microbiol. 2017;55(2):96-103. Published online January 26, 2017; DOI: https://doi.org/10.1007/s12275-017-6300-3

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Abstract

To clarify the interactions between a common food spoilage yeast and two pathogenic bacteria involved in outbreaks associated with fruit juices, the present paper studies the effect of the interplay of Candida krusei, collected from UF membranes, with Escherichia coli O157:H7 and Salmonella enterica in the overall process of adhesion and colonization of abiotic surfaces. Two different cases were tested: a) co-adhesion by pathogenic bacteria and yeasts, and b) incorporation of bacteria to pre-adhered C. krusei cells. Cultures were made on stainless steel at 25°C using apple juice as culture medium. After 24 h of co-adhesion with C. krusei, both E. coli O157:H7 and S. enterica increased their counts 1.05 and 1.11 log CFU cm2, respectively. Similar increases were obtained when incorporating bacteria to pre-adhered cells of Candida. Nevertheless C. krusei counts decreased in both experimental conditions, in a) 0.40 log CFU cm2 and 0.55 log CFU cm2 when exposed to E. coli O157:H7 and S. enterica and in b) 0.18 and 0.68 log CFU cm2, respectively. This suggests that C. krusei, E. coli O157:H7, and S. enterica have a complex relationship involving physical and chemical interactions on food contact surfaces. This study supports the possibility that pathogen interactions with members of spoilage microbiota, such as C. krusei, might play an important role for the survival and dissemination of E. coli O157:H7 and Salmonella enterica in food-processing environments. Based on the data obtained from the present study, much more attention should be given to prevent the contamination of these pathogens in acidic drinks.

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Effectiveness of sodium hypochlorite and benzalkonium chloride in reducing spoilage yeast biofilms on food contact surfaces
Manuel Alejandro Palencia Díaz, María Clara Tarifa, Patricia Liliana Marucci, Diego Bautista Genovese, Lorena Inés Brugnoni
Biofouling.2024; 40(10): 964. CrossRef
Application of natamycin and farnesol as bioprotection agents to inhibit biofilm formation of yeasts and foodborne bacterial pathogens in apple juice processing lines
María del Rosario Agustín, María Clara Tarifa, María Soledad Vela-Gurovic, Lorena Inés Brugnoni
Food Microbiology.2023; 109: 104123. CrossRef
Candida krusei is the major contaminant of ultrafiltration and reverse osmosis membranes used for cranberry juice production
Sherazade Fikri, Marie-Hélène Lessard, Véronique Perreault, Alain Doyen, Steve Labrie
Food Microbiology.2023; 109: 104146. CrossRef
Application of Natamycin and Farnesol as Biocontrol Agents of Multi-Species Biofilms on Industrial Surfaces in Apple Juice
María del Rosario Agustín, Maria Clara Tarifa, Maria Soledad Vela-Gurovic, Lorena Ines Brugnoni
SSRN Electronic Journal .2022;[Epub] CrossRef
Enhanced microbial inactivation by carbon dioxide through mechanical effects
Ratka Hoferick, Angelos Ntovas, Qasim Alhusaini, Mareike Müller, Stéphan Barbe, Holger Schönherr
The Journal of Supercritical Fluids.2021; 175: 105273. CrossRef
Yeast biofilm in food realms: occurrence and control
Giacomo Zara, Marilena Budroni, Ilaria Mannazzu, Francesco Fancello, Severino Zara
World Journal of Microbiology and Biotechnology.2020;[Epub] CrossRef
Disinfection efficacy over yeast biofilms of juice processing industries
María C. Tarifa, Jorge E. Lozano, Lorena I. Brugnoni
Food Research International.2018; 105: 473. CrossRef
Identification des espèces de levures isolées de l’attiéké commercialisé sur les marchés à Abidjan (Côte d’Ivoire) : étude préliminaire
C.G. Kouadio-Yapo, G.S.P. Dou, N.A.D. Aka, K.D. Zika, K.D. Adoubryn, M. Dosso
Journal de Mycologie Médicale.2018; 28(2): 305. CrossRef
Multispecies biofilms between Listeria monocytogenes and Listeria innocua with resident microbiota isolated from apple juice processing equipment
María del Rosario Agustín, Lorena Brugnoni
Journal of Food Safety.2018;[Epub] CrossRef

Research Support, Non-U.S. Gov't

Inactivation of Barotolerant Strains of Listeria monocytogenes and Escherichia coli O157:H7 by Ultra High Pressure and tert-Butylhydroquinone Combination: Yoon-Kyung Chung , Ahmed E. Yousef; J. Microbiol. 2008;46(3):289-294. Published online July 5, 2008; DOI: https://doi.org/10.1007/s12275-008-0090-6

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Abstract: Antimicrobial efficacy of ultra-high-pressure (UHP) can be enhanced by application of additional hurdles. The objective of this study was to systematically assess the enhancement in pressure lethality by TBHQ treatment, against barotolerant strains of Escherichia coli O157:H7 and Listeria monocytogenes. Two L. monocytogenes Scott A and the barotolerant OSY-328 strain, and two E. coli O157:H7 strains, EDL-933 and its barotolerant mutant, OSY-ASM, were tested. Cell suspensions containing TBHQ (50 ppm, dissolved in dimethyl sulfoxide) were pressurized at 200 to 500 MPa (23±2°C) for 1 min, plated on tryptose agar and enumerated the survivors. The TBHQ-UHP combination resulted in synergistic inactivation of both pathogens, with different degrees of lethality among strains. The pressure lethality threshold, for the combination treatment, was lower for E. coli O157:H7 (≥ 200 MPa) than for L. monocytogenes (> 300 MPa). E. coli O157:H7 strains were extremely sensitive to the TBHQ-UHP treatment, compared to Listeria strains. Interestingly, a control treatment involving DMSO-UHP combination consistently resulted in higher inactivation than that achieved by UHP alone, against all strains tested. However, sensitization of the pathogens to UHP by the additives (TBHQ in DMSO) was prominently greater for UHP than DMSO. Differences in sensitivities to the treatment between these two pathogens may be attributed to discrepancies in cellular structure or physiological functions.

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