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Kyrkou, I.

Publications and source records attributed to Kyrkou, I..

3 recordsLinked to original sources

Detecting transcriptional responses and comparing the virulence of Pseudomonas aeruginosa cystic fibrosis isolates in a mung bean model

A mung bean infection model has previously been shown to differentiate between non-virulent and virulent Pseudomonas aeruginosa bacteria. However, it remains unclear how plant and bacteria adjust their gene expression during infection and whether the mung bean model can be used to compare the virulence of clinical cystic fibrosis (CF) P. aeruginosa lung isolates. Here, we first explored temporal transcriptomics of P. aeruginosa PAO1 and mung bean during an infection. We found that bacterial gene expression followed temporal changes, with an increase in the expression of O-antigen biosynthetic genes, chemotaxis, phosphate intake and phenazine production. Mung bean responded by upregulating genes associated with defence mechanisms and downregulating genes involved in the plant development. From the PAO1 perspective, the core-transcriptomic responses in the mung bean were similar to its responses previously observed in wound and excision and in in vitro media and sputum models, while differed from those observed in the bronchial cell model. Furthermore, we used the mung bean to assess the virulence of 119 clinical P. aeruginosa CF strains originating from the Copenhagen CF clinic. By quantifying bacterial virulence as a reduction in root and shoot growth and weight of the seeds, we found that CF strains isolated at later compared to early stages of lung infections showed higher virulence. This difference corresponded with the higher number of immune modulation-associated virulence genes and lower number of motility and effector genes, present in the genomes of late compared to early isolated CF strains. IMPORTANCEOur results demonstrate that based on PAO1 transcriptional profile, the mung bean model is similar to in vitro and wound infection models but differs from cell and bronchial models. Moreover, the mung bean model can detect virulence differences between clinical P. aeruginosa CF strains, making it a potentially useful high-throughput in vivo model for bacterial virulence screening.

microbiology↗

Isolation and characterisation of novel phages infecting Lactobacillus plantarum and proposal of a new genus, \"Silenusvirus\".

Bacteria of Lactobacillus sp. are very useful to humans. However, the biology and genomic diversity of their (bacterio)phage enemies remains understudied. Knowledge on Lactobacillus phage diversity should broaden to develop efficient phage control strategies. To this end, organic waste samples were screened for phages against two wine-related Lactobacillus plantarum strains. Isolates were shotgun sequenced and compared against the phage database and each other by phylogenetics and comparative genomics. The new isolates had only three distant relatives from the database but displayed a high overall degree of genomic similarity amongst them. The latter allowed for the use of one isolate as a representative to conduct transmission electron microscopy and structural protein sequencing, and to study phage adsorption and growth kinetics. The microscopy and proteomics tests confirmed the observed diversity of the new isolates and supported their classification to the family Siphoviridae and the proposal of the new phage genus \"Silenusvirus\".

microbiology↗

Seasonal microbial dynamics on grapevine leaves under biocontrol and copper fungicide treatments

Winemakers have long used copper as a fungicide on grapevine. However, the potential of copper to accumulate on soil and affect the biota poses a challenge to achieving sustainable agriculture. One recently developed option is the use of biocontrol agents to replace or complement traditional methods. In the present study, a field experiment was conducted in South Africa in which the leaves in two blocks of a vineyard were periodically treated with either copper sulphate or sprayed with Lactobacillus plantarum MW-1 as a biocontrol agent. This study evaluated the impact of the two treatments on the bacterial and fungal communities as they changed during the growing season. To do this, NGS was combined with quantitative strain-specific and community qPCRs. The results revealed the progression of the microbial communities throughout the season and how the different treatments affected the microbiota. Bacteria appeared to be relatively stable at the different time points, with the only taxa that systematically changed between treatments being Lactobacillaceae, which included reads from the biocontrol agent. Cells of Lactobacillus plantarum MW-1 were only detected on treated leaves using strain-specific qPCR, with its amount spanning from 103to 105cells/leaves. Conversely the fungal community was largely shaped by a succession of different dominant taxa over the months. Between treatments, only a few fungal taxa appeared to change significantly and the number of ITS copies was also comparable. In this regards, the two treatments seemed to affect the microbial community similarly, revealing the potential of this biocontrol strain as a promising alternative among sustainable fungicide treatments, although further investigation is required.

microbiology↗