Jayson J. A. Rose

The obligate necrotrophic parasite, Candidatus Mycosynbacter amalyticus, a member of the Patescibacteria has been isolated from wastewater. Subsequent efforts have been directed toward unravelling its biological lifecycle and attachment mechanism facilitating infection and subsequent lysis of its Actinobacterial host, Gordonia amarae. Here, using electron cryo-tomography (CryoET), we reveal the molecular anatomy of parasitic Mycosynbacter amalyticus cells, uncovering an unusual infection process. Through laboratory-based evolution experiments, we generated eleven slow-growing independent spontaneous Gordonia amarae resistant mutants. Mycolic acids (MA) are key components of the outer cellular envelope of G. amarae and other Actinobacteria, with MA being the physical attribute implicated in G. amarae associated wastewater foaming. CryoET and genome sequencing exposed absence of intact MA and an associated suite of mutations predominantly occurring within the pks13 and pptT genes of the MA biosynthetic pathway. Our findings suggest that MA structural integrity is critical for attachment of Ca. Mycosynbacter amalyticus to its host.

Pantoea stewartii , a plant pathogen, is primarily transmitted through contaminated seeds and insect vectors, with the corn flea beetle (Chaetocnema pulicaria) being the primary carrier. P. stewartii is a bacterium belonging to the order Enterobacterales and can lead to crop diseases that have a significant economic impact worldwide. Due to its high potential for spread, P. stewartii is classified as a quarantine organism in numerous countries. Despite its impact on agriculture, the limited genome sequences of P. stewartii hamper understanding of its pathogenicity and host specificity, and the development of effective control strategies. In this study, a P. stewartii strain (C10109_Jinnung) was discovered in the faecal matter of the Critically Endangered western ground parrot/kyloring (Pezoporus flaviventris) in Australia, which to our knowledge is the first reported P. stewartii genome from a bird source. Whole-genome sequencing and phylogenomic analysis of strain C10109_Jinnung, obtained from a captive psittacine, provides new insights into the genetic diversity and potential transmission route for the spread of P. stewartii beyond insects and plants, where P. stewartii is typically studied. Our findings provide new insights into the potential transmission route for spread of P. stewartii and expand the known transmission agents beyond insects and plants. Expanding the catalogue of P. stewartii genomes is fundamental to improving understanding of the pathogenicity, evolution and dissemination, and to develop effective control strategies to reduce the substantial economic losses associated with P. stewartii in various crops and the potential impact of endangered animal species.

Soil contains an enormous diversity of microorganisms and can act as a reservoir of antibiotic resistance determinants. This study identified and compared the bacterial diversity and the antimicrobial resistance profile of clinically-relevant isolates around a newly developed hospital and university precinct. Eight soil samples were collected, genomic DNA was extracted and 16S rRNA gene sequencing was performed. Bacterial isolates cultured from the soil were identified using MALDI-TOF. Antibiotic sensitivity testing (AST) was performed on a subset of isolates. The soil from both precincts were similarly diverse. Phylum Proteobacteria was prevalent in all samples and was the most abundant in one of the hospital sites. Cyanobacteria was abundant in two hospital sites closer to a sewage treatment plant. Bacterial diversity was only significantly different between two of the hospital sites. A total of 22 Gram-negative organisms were isolated by culture. AST revealed that the soil isolates from both precincts exhibited low resistance. The unidentified bacteria closer to the hospital precinct with human interactions possibly hints at the role of anthropogenic activities on the soil microbial diversity. The abundance of Proteobacteria (causing majority of human infections) and Cyanobacteria nearer to the hospital premises, comprising more immunocompromised and immunocompetent individuals, is concerning.

The production of a stable foam on the surfaces of reactors is a global operating problem in activated sludge plants. In many cases, these foams are stabilized by hydrophobic members of the Mycolata, a group of Actinobacteria whose outer membranes contain long-chain hydroxylated mycolic acids. There is currently no single strategy which works for all foams. One attractive approach is to use lytic bacteriophages specific for the foam stabilizing Mycolata population. Such phages are present in activated sludge mixed liquor and can be recovered readily from it. However, no phage has been recovered which lyses Gordonia amarae and Gordonia pseudoamarae, probably the most common foaming Mycolata members. Whole genome sequencing revealed that both G. amarae and G. pseudoamarae from plants around the world are particularly well endowed with genes encoding antiviral defence mechanisms. However, both these populations were lysed rapidly by a parasitic nanobacterium isolated from a plant in Australia. This organism, a member of the Saccharibacteria, was also effective against many other Mycolata, thus providing a potential agent for control of foams stabilized by them.

Many wastewater treatment plants around the world suffer from the operational problem of foaming. This is characterized by a persistent stable foam that forms on the aeration basin, which reduces effluent quality. The foam is often stabilized by a highly hydrophobic group of Actinobacteria known as the Mycolata¹. Gordonia amarae is one of the most frequently reported foaming members¹. With no currently reliable method for treating foams, phage biocontrol has been suggested as an attractive treatment strategy². Phages isolated from related foaming bacteria can destabilize foams at the laboratory scale^3,4; however, no phage has been isolated that lyses G. amarae. Here, we assemble the complete genomes of G. amarae and a previously undescribed species, Gordonia pseudoamarae, to examine mechanisms that encourage stable foam production. We show that both of these species are recalcitrant to phage infection via a number of antiviral mechanisms including restriction, CRISPR–Cas and bacteriophage exclusion. Instead, we isolate and cocultivate an environmental ultrasmall epiparasitic bacterium from the phylum Saccharibacteria that lyses G. amarae and G. pseudoamarae and several other Mycolata commonly associated with wastewater foams. The application of this parasitic bacterium, ‘Candidatus Mycosynbacter amalyticus’, may represent a promising strategy for the biocontrol of bacteria responsible for stabilizing wastewater foams.

Bacteriophages (phages) are biological entities that have attracted a great deal of attention in recent years. They have been reported as the most abundant biological entities on the planet and their ability to impact the composition of bacterial communities is of great interest. In this review, we aim to explore where phages exist in natural and artificial environments and how they impact communities. The natural environment in this review will focus on the human body, soils, and the marine environment. In these naturally occurring environments there is an abundance of phages suggesting a role in the maintenance of bacterial community homeostasis. The artificial environment focuses on wastewater treatment plants, industrial processes, followed by pharmaceutical formulations. As in natural environments, the existence of bacteria in manmade wastewater treatment plants and industrial processes inevitably attracts phages. The presence of phages in these environments can inhibit the bacteria required for efficient water treatment or food production. Alternatively, they can have a positive impact by eliminating recalcitrant organisms. Finally, we conclude by describing how phages can be manipulated or formulated into pharmaceutical products in the laboratory for use in natural or artificial environments.