Metataxonomic Analysis of Bacteria Entrapped in a Stalactite’s Core and Their Possible Environmental Origins

Much is known about microbes originally identified in caves, but little is known about the entrapment of microbes (bacteria) in stalactites and their possible environmental origins. This study presents data regarding the significant environmental distribution of prokaryotic bacterial taxa of a Greek stalactite core. We investigated the involvement of those bacteria communities in stalactites using a metataxonomic analysis approach of partial 16S rRNA genes. The metataxonomic analysis of stalactite core material revealed an exceptionally broad ecological spectrum of bacteria classified as members of Proteobacteria, Actinobacteria, Firmicutes, Verrucomicrobia, and other unclassified bacteria. We concluded that (i) the bacterial transport process is possible through water movement from the upper ground cave environment, forming cave speleothems such as stalactites, (ii) bacterial genera such as Polaromonas, Thioprofundum, and phylum Verrucomicrobia trapped inside the stalactite support the paleoecology, paleomicrobiology, and paleoclimate variations, (iii) the entrapment of certain bacteria taxa associated with water, soil, animals, and plants such as Micrococcales, Propionibacteriales, Acidimicrobiales, Pseudonocardiales, and α-, β-, and γ-Proteobacteria.

Evidence for biosurfactant-induced flow in corners and bacterial spreading in unsaturated porous media

The spread of pathogenic bacteria in unsaturated porous media, where air and liquid coexist in pore spaces, is the major cause of soil contamination by pathogens, soft rot in plants, food spoilage, and many pulmonary diseases. However, visualization and fundamental understanding of bacterial transport in unsaturated porous media are currently lacking, limiting the ability to address the above contamination- and disease-related issues. Here, we demonstrate a previously unreported mechanism by which bacterial cells are transported in unsaturated porous media. We discover that surfactant-producing bacteria can generate flows along corners through surfactant production that changes the wettability of the solid surface. The corner flow velocity is on the order of several millimeters per hour, which is the same order of magnitude as bacterial swarming, one of the fastest known modes of bacterial surface translocation. We successfully predict the critical corner angle for bacterial corner flow to occur based on the biosurfactant-induced change in the contact angle of the bacterial solution on the solid surface. Furthermore, we demonstrate that bacteria can indeed spread by producing biosurfactants in a model soil, which consists of packed angular grains. In addition, we demonstrate that bacterial corner flow is controlled by quorum sensing, the cell–cell communication process that regulates biosurfactant production. Understanding this previously unappreciated bacterial transport mechanism will enable more accurate predictions of bacterial spreading in soil and other unsaturated porous media.

Soil protists can actively redistribute beneficial bacteria along Medicago truncatula roots

The rhizosphere is the region of soil directly influenced by plant roots. The microbial community in the rhizosphere includes fungi, protists, and bacteria, all of which play a significant role in plant health. The beneficial bacterium Sinorhizobium meliloti infects growing root hairs on nitrogen starved leguminous plants. Infection leads to the formation of a root nodule, where S. meliloti converts atmospheric nitrogen to ammonia, the usable form of nitrogen for plants. However, S. meliloti, often found in biofilms, travels slowly; whereas infectible root hairs are found at the growing root tip, potentially causing many root hairs to remain uninfected by S. meliloti when it is delivered as a seed inoculant. Soil protists are an important component of the rhizosphere system who prey on soil bacteria and have been known to egest undigested phagosomes. We show that the soil protist, Colpoda sp., plays an important role in transporting S. meliloti down Medicago truncatula roots. By using pseudo-3D soil microcosms we directly observed the presence of fluorescently labelled S. meliloti along M. truncatula roots and track the displacement of bacteria over time. In the presence of Colpoda sp., S. meliloti was detected 44 mm, on average, farther down the roots, compared with the Bacteria Only Treatment. Facilitating bacterial transport may be an important mechanism whereby soil protists promote plant health. Protist facilitated transport as a sustainable agriculture biotechnology has the potential to boost efficacy of bacterial inoculants, avoid overuse of nitrogen fertilizers, and enhance performance of no-till farming practices.

Comparative Transport of Legionella and E. coli through Saturated Porous Media in a Two-Dimensional Tank

This study investigated bacterial transport in a two-dimensional (2-D) tank to evaluate the bacterial behavior of Legionella pneumophila as compared to Escherichia coli under saturated flow to simulate aquifer conditions. The experiments were performed in a 2-D tank packed with 3700 in3 (60,632 cm3) of commercially available bagged play sand under saturated conditions. The tank was disinfected by backwashing with 10% chlorine solution and subsequently neutralized by backwashing with tap water containing sodium thiosulphate (Na2S2O3) to ensure no chlorine residual. Bacterial transport was measured using samples collected from ports located at vertical transport distances of 5, 15 and 25 inches (12.7, 38.1 and 63.5 cm, respectively) below the sand surface along two vertical sections in the tank. An influent concentration of 105 CFU/mL was used for bacterial cells and the vertical fluid transport rate was 10.3 in/day (26.2 cm/day), controlled using a peristaltic pump at the bottom outlet. Legionella breakthroughs were recorded at 8, 22 and 35 h for the ports on the right side and 9, 24 and 36 h for the ports on the left side, at 5, 15 and 25 inch depths, respectively. At the same depths, E. coli breakthroughs were recorded at 5, 17 and 30 h for the ports on the right side and 7, 19 and 31 h for the ports on the left sides. The delay in Legionella transport compared to E. coli is homologous to Legionella’s pleomorphic nature. This study provides evidence of the mobility of both E. coli and Legionella in saturated aquifer conditions at a scale more representative of actual aquifer conditions. This study also provides a substantive basis for the premise that cell characteristics affect transport characteristics under those conditions.

A Kayvirus Distant Homolog of Staphylococcal Virulence Determinants and VISA Biomarker Is a Phage Lytic Enzyme

Staphylococcal bacteriophages of the Kayvirus genus are candidates for therapeutic applications. One of their proteins, Tgl, is slightly similar to two staphylococcal virulence factors, secreted autolysins of lytic transglycosylase motifs IsaA and SceD. We show that Tgl is a lytic enzyme secreted by the bacterial transport system and localizes to cell peripheries like IsaA and SceD. It causes lysis of E. coli cells expressing the cloned tgl gene, but could be overproduced when depleted of signal peptide. S. aureus cells producing Tgl lysed in the presence of nisin, which mimics the action of phage holin. In vitro, Tgl protein was able to destroy S. aureus cell walls. The production of Tgl decreased S. aureus tolerance to vancomycin, unlike the production of SceD, which is associated with decreased sensitivity to vancomycin. In the genomes of kayviruses, the tgl gene is located a few genes away from the lysK gene, encoding the major endolysin. While lysK is a late phage gene, tgl can be transcribed by a host RNA polymerase, like phage early genes. Taken together, our data indicate that tgl belongs to the kayvirus lytic module and encodes an additional endolysin that can act in concert with LysK in cell lysis.