Genome-Scale Analysis of Mycoplasma agalactiae Loci Involved in Interaction with Host Cells

Mycoplasma agalactiae is an important pathogen of small ruminants, in which it causes contagious agalactia. It belongs to a large group of “minimal bacteria” with a small genome and reduced metabolic capacities that are dependent on their host for nutrients. Mycoplasma survival thus relies on intimate contact with host cells, but little is known about the factors involved in these interactions or in the more general infectious process. To address this issue, an assay based on goat epithelial and fibroblastic cells was used to screen a M. agalactiae knockout mutant library. Mutants with reduced growth capacities in cell culture were selected and 62 genomic loci were identified as contributing to this phenotype. As expected for minimal bacteria, “transport and metabolism” was the functional category most commonly implicated in this phenotype, but 50% of the selected mutants were disrupted in coding sequences (CDSs) with unknown functions, with surface lipoproteins being most commonly represented in this category. Since mycoplasmas lack a cell wall, lipoproteins are likely to be important in interactions with the host. A few intergenic regions were also identified that may act as regulatory sequences under co-culture conditions. Interestingly, some mutants mapped to gene clusters that are highly conserved across mycoplasma species but located in different positions. One of these clusters was found in a transcriptionally active region of the M. agalactiae chromosome, downstream of a cryptic promoter. A possible scenario for the evolution of these loci is discussed. Finally, several CDSs identified here are conserved in other important pathogenic mycoplasmas, and some were involved in horizontal gene transfer with phylogenetically distant species. These results provide a basis for further deciphering functions mediating mycoplasma-host interactions

Deep Sequencing Whole Transcriptome Exploration of the σE Regulon in Neisseria meningitidis

Bacteria live in an ever-changing environment and must alter protein expression promptly to adapt to these changes and survive. Specific response genes that are regulated by a subset of alternative σ70-like transcription factors have evolved in order to respond to this changing environment. Recently, we have described the existence of a σE regulon including the anti-σ-factor MseR in the obligate human bacterial pathogen Neisseria meningitidis. To unravel the complete σE regulon in N. meningitidis, we sequenced total RNA transcriptional content of wild type meningococci and compared it with that of mseR mutant cells (ΔmseR) in which σE is highly expressed. Eleven coding genes and one non-coding gene were found to be differentially expressed between H44/76 wildtype and H44/76ΔmseR cells. Five of the 6 genes of the σE operon, msrA/msrB, and the gene encoding a pepSY-associated TM helix family protein showed enhanced transcription, whilst aniA encoding a nitrite reductase and nspA encoding the vaccine candidate Neisserial surface protein A showed decreased transcription. Analysis of differential expression in IGRs showed enhanced transcription of a non-coding RNA molecule, identifying a σE dependent small non-coding RNA. Together this constitutes the first complete exploration of an alternative σ-factor regulon in N. meningitidis. The results direct to a relatively small regulon indicative for a strictly defined response consistent with a relatively stable niche, the human throat, where N. meningitidis resides

Effect of Ulvan on the Biocontrol Activity of <i>Debaryomyces hansenii</i> and <i>Stenotrophomonas rhizophila</i> against Fruit Rot of <i>Cucumis melo</i> L.

In the present study, the following was investigated: (a) The effect of ulvan on in vivo and in vitro biocontrol of Debaryomyces hansenii and Stenotrophomonas rhizophila against Fusarium proliferaum and (b) the effect of ulvan on in vivo and in vitro growth of D. hansenii and S. rhizophila and muskmelon quality parameters. The results showed that the biocontrol activity of D. hansenii and S. rhizophila could be enhanced by ulvan (5 g/L). The combination of ulvan and S. rhizophila resulted in a more effective control of fruit rot in comparison to fungicide benomyl. On in vitro growth of F. proliferatum, individual treatments of D. hansenii and S. rhizophila inhibited spore germination and mycelial growth with no statistical difference with the combined treatments. Ulvan does not have a direct effect on the in vivo and in vitro growth of D. hansenii and S. rhizophila. Furthermore, the combined treatments improve the natural disease incidence and quality parameters like weight, firmness, total soluble solids (TSS), and pH. These results suggest that the use of ulvan may be an effective method to improve the biological activity of D. hansenii and S. rhizophila

Biocontrol of Postharvest Fruit Fungal Diseases by Bacterial Antagonists: A Review

This review deals with the main mechanisms of action exerted by antagonistic bacteria, such as competition for space and nutrients, suppression via siderophores, hydrolytic enzymes, antibiosis, biofilm formation, and induction of plant resistance. These mechanisms inhibit phytopathogen growth that affects postharvest fruit since quality and safety parameters are influenced by the action of these microorganisms, which cause production losses in more than 50% of fruit tree species. The use of synthetic fungicide products has been the dominant control strategy for diseases caused by fungi. However, their excessive and inappropriate use in intensive agriculture has brought about problems that have led to environmental contamination, considerable residues in agricultural products, and phytopathogen resistance. Thus, there is a need to generate alternatives that are safe, ecological, and economically viable to face this problem. Phytopathogen inhibition in fruit utilizing antagonist microorganisms has been recognized as a type of biological control (BC), which could represent a viable and environmentally safe alternative to synthetic fungicides. Despite the ecological benefit that derives from the use of controllers and biological control agents (BCA) at a commercial level, their application and efficient use has been minimal at a global level