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Dynamic social interactions and keystone species shape the diversity and stability of mixed-species biofilms – an example from dairy isolates

Identifying interspecies interactions in mixed-species biofilms is a key challenge in microbial ecology and is of paramount importance given that interactions govern community functionality and stability. We previously reported a bacterial four-species biofilm model comprising Stenotrophomonas rhizophila, Bacillus licheniformis, Microbacterium lacticum, and Calidifontibacter indicus that were isolated from the surface of a dairy pasteuriser after cleaning and disinfection. These bacteria produced 3.13-fold more biofilm mass compared to the sum of biofilm masses in monoculture. The present study confirms that the observed community synergy results from dynamic social interactions, encompassing commensalism, exploitation, and amensalism. M. lacticum appears to be the keystone species as it increased the growth of all other species that led to the synergy in biofilm mass. Interactions among the other three species (in the absence of M. lacticum) also contributed towards the synergy in biofilm mass. Biofilm inducing effects of bacterial cell-free-supernatants were observed for some combinations, revealing the nature of the observed synergy, and addition of additional species to dual-species combinations confirmed the presence of higher-order interactions within the biofilm community. Our findings provide understanding of bacterial interactions in biofilms which can be used as an interaction–mediated approach for cultivating, engineering, and designing synthetic bacterial communities.

Legacies of temperature fluctuations promote stability in marine biofilm communities

The increasing frequency and intensity of extreme climate events are driving significant biodiversity shifts across ecosystems. Yet, the extent to which these climate legacies will shape the response of ecosystems to future perturbations remains poorly understood. Here, we tracked taxon and trait dynamics of rocky intertidal biofilm communities under contrasting regimes of warming (fixed vs. fluctuating) and assessed how they influenced stability dimensions in response to temperature extremes. Fixed warming enhanced the resistance of biofilm by promoting the functional redundancy of stress-tolerance traits. In contrast, fluctuating warming boosted recovery rate through the selection of fast-growing taxa at the expense of functional redundancy. This selection intensified a trade-off between stress tolerance and growth further limiting the ability of biofilm to cope with temperature extremes. Anticipating the challenges posed by future extreme events, our findings offer a forward-looking perspective on the stability of microbial communities in the face of ongoing climatic change.

Exploration of vanoxerine analogues as antibacterial agents

Mycobacterium tuberculosis is a bacterial pathogen, responsible for approximately 1.3 million deaths in 2022 through tuberculosis infections. The complex treatment regimen required to treat tuberculosis and growing rates of drug resistance, necessitates the development of new anti-mycobacterial agents. One approach is to repurpose drugs from other clinical applications. Vanoxerine (GBR 12909) was previously shown to have anti-mycobacterial activity, through dissipating the membrane electric potential and hence, cellular energetics. Several vanoxerine analogues were synthesised in this study, which exhibited a range of activities against mycobacteria and enterococcus. All active analogues had similar impacts on the membrane electric potential and inhibition of ethidium bromide efflux. The most active compound displayed reduced inhibitory activity against the known human target of vanoxerine, the dopamine transporter. This work has identified a promising analogue, which could provide a starting point for further medicinal chemistry and drug development efforts to target mycobacteria.

Genomic and transcriptomic insights into complex virus–prokaryote interactions in marine biofilms

Marine biofilms are complex communities of microorganisms that play a crucial ecological role in oceans. Although prokaryotes are the dominant members of these biofilms, little is known about their interactions with viruses. By analysing publicly available and newly sequenced metagenomic data, we identified 2446 virus–prokaryote connections in 84 marine biofilms. Most of these connections were between the bacteriophages in the Uroviricota phylum and the bacteria of Proteobacteria, Cyanobacteria and Bacteroidota. The network of virus–host pairs is complex; a single virus can infect multiple prokaryotic populations or a single prokaryote is susceptible to several viral populations. Analysis of genomes of paired prokaryotes and viruses revealed the presence of 425 putative auxiliary metabolic genes (AMGs), 239 viral genes related to restriction–modification (RM) systems and 38,538 prokaryotic anti-viral defence-related genes involved in 15 defence systems. Transcriptomic evidence from newly established biofilms revealed the expression of viral genes, including AMGs and RM, and prokaryotic defence systems, indicating the active interplay between viruses and prokaryotes. A comparison between biofilms and seawater showed that biofilm prokaryotes have more abundant defence genes than seawater prokaryotes, and the defence gene composition differs between biofilms and the surrounding seawater. Overall, our study unveiled active viruses in natural biofilms and their complex interplay with prokaryotes, which may result in the blooming of defence strategists in biofilms. The detachment of bloomed defence strategists may reduce the infectivity of viruses in seawater and result in the emergence of a novel role of marine biofilms.

Dynamic and intricate regulation by the Csr sRNAs in the Arctic Pseudoalteromonas fuliginea

The Csr (Carbon Storage Regulator) system is pivotal in controlling various cellular functions in most bacteria, primarily through the CsrA protein and its antagonistic sRNAs. However, riboregulatory networks are less explored in non-model organisms, particularly those in extreme environments. In this study, we discovered two new sRNAs of the Csr system, Pf2 and Pf3, in the Arctic bacterium Pseudoalteromonas fuliginea BSW20308, along with the previously known Pf1. By studying the impact of these Pf sRNAs on CsrA targetomes and physiological processes, we found a significant influence on various cellular functions and a collective effect on the interaction dynamics between CsrA and RNAs. Furthermore, we identified additional sRNAs that can interact with CsrA and mRNAs. Overall, our results emphasize the growing influence of the Csr system on cellular physiology through intricate sRNA regulation of CsrA, revealing riboregulatory network complexity and significance in non-model organisms.

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