Microbial protein metabolism in the monogastric gastrointestinal tract: a review

Dietary and endogenous protein that become available for the microbiota in the hindgut can be metabolized via different routes. They can become building blocks for the microbial cells or enter different catabolic pathways. Protein degradation via fermentation pathways is seen as a non-preferred route as it results in the formation and release of metabolites that can interfere with biological systems in the host and can have deleterious outcomes. Reducing protein fermentation and guiding the metabolism towards less toxic end-products might be possible targets for improving host health. To do so, more knowledge on factors manipulating the process of microbial protein metabolism, including on substrate availability, microbial composition and segmental differences in the hindgut, is required.

Bionanotechnology Approaches to Combat Biofilms and Drug Resistance

This chapter deals with the formation of biofilms, their resistance to antibacterial agents, the importance and risk of biofilms, and nanotechnology methods for biofilm control in the food industry. Biofilm is a multi-layer cell cluster embedded in an organic polymer matrix, which protects microbial cells from environmental stress, antibiotics, and disinfectants. Microorganisms that live in contact points and the environment in food processing are mostly harmful because the microbial community in the wrong location can lead to contamination of the surfaces and products produced during the processing. When new nanomaterials (for example, silver or copper are incorporated) are used, the growth of surface biofilms can also be reduced. In recent years, new nanotechnology-based antimicrobials have been designed to kill planktonic, antibiotic-resistant bacteria, but additional requirements rather than the mere killing of suspended bacteria must be met to combat biofilm-infections.

Illuminating the Biochemical Interaction of Antimicrobial Few-Layer Black Phosphorus with Microbial Cells Using Synchrotron macro-ATR-FTIR

In the fight against drug-resistant pathogenic bacterial and fungal cells, low-dimensional materials are emerging as a promising alternative treatment method. Specifically, few-layer black phosphorus (BP) has demonstrated its effectiveness against...

Microbial associations of shallow-water Mediterranean marine cave Solenogastres (Mollusca)

The first cave-dwelling Solenogastres—marine shell-less worm-like mollusks—were sampled from Mediterranean marine caves floor silt in the Marseille area. The mollusks were 1.5 mm in length, had a transparent body with shiny spicules and appear to represent a new Tegulaherpia species. Electron microscopy revealed a high number of microbial cells, located on the surface of the spicules as well as in the cuticle of Tegulaherpia sp. The observed microbial cells varied in morphology and were unequally distributed through the cuticle, reaching a highest density on the dorsal and lateral sides and being practically absent on the ventral side. Next Generation Sequencing (NGS) of V4 region of 16S rRNA gene amplicons, obtained from the DNA samples of whole bodies of Tegulaherpia sp. revealed three dominating microorganisms, two of which were bacteria of Bacteroidetes and Nitrospirae phyla, while the third one represented archaea of Thaumarchaeota phylum. The Operational Taxonomic Unit (OTU), affiliated with Bacteroidetes was an uncultured bacteria of the family Saprospiraceae (93–95% of Bacteroidetes and 25–44% of the total community, depending on sample), OTU, affiliated with Nitrospirae belonged to the genus Nitrospira (8–30% of the community), while the thaumarchaeal OTU was classified as Candidatus Nitrosopumilus (11–15% of the community). Members of these three microbial taxa are known to form associations with various marine animals such as sponges or snails where they contribute to nitrogen metabolism or the decomposition of biopolymers. A similar role is assumed to be played by the microorganisms associated with Tegulaherpia sp.

Calculation and Interpretation of Substrate Assimilation Rates in Microbial Cells Based on Isotopic Composition Data Obtained by nanoSIMS

Stable isotope probing (SIP) combined with nano-scale secondary ion mass spectrometry (nanoSIMS) is a powerful approach to quantify assimilation rates of elements such as C and N into individual microbial cells. Here, we use mathematical modeling to investigate how the derived rate estimates depend on the model used to describe substrate assimilation by a cell during a SIP incubation. We show that the most commonly used model, which is based on the simplifying assumptions of linearly increasing biomass of individual cells over time and no cell division, can yield underestimated assimilation rates when compared to rates derived from a model that accounts for cell division. This difference occurs because the isotopic labeling of a dividing cell increases more rapidly over time compared to a non-dividing cell and becomes more pronounced as the labeling increases above a threshold value that depends on the cell cycle stage of the measured cell. Based on the modeling results, we present formulae for estimating assimilation rates in cells and discuss their underlying assumptions, conditions of applicability, and implications for the interpretation of intercellular variability in assimilation rates derived from nanoSIMS data, including the impacts of storage inclusion metabolism. We offer the formulae as a Matlab script to facilitate rapid data evaluation by nanoSIMS users.

Synergistic Effect of Ultrasound Cavitation and Gas in the Water Disinfection

The paper considers water purification processes from Bacillus bacteria type under the conditions of gases bubbling only (argon, helium, oxygen, and carbon dioxide), cavitation and combined action of gas and cavitation. The synergistic effect was found under conditions of simultaneous action of gas and cavitation (kd(gas/US ) >kd(gas) + kd(US) almost double) and it was shown that kd(gas/US) >kd(gas) by almost an order of magnitude. Relative series of effective destruction of microbial cells was established: Ar/US > О2/US >Не/US > СО2/US. Destruction degree of the cells reaches 70 %at the short-term Ar/US exposure (~8 min), which is 7 times more active than cavitation action and 13.5 times more than bubbling of Aralone.

Isolating and culturing of single microbial cells by laser ejection sorting technology

Single cell isolation and cultivation play an important role in studying physiology, gene expression and functions of microorganisms. A series of single-cell isolation technologies have been developed, among which single-cell ejection technology is one of the most promising. Single cell ejection technology has applied Laser Induced Forward Transfer Technique (LIFT) to isolate bacteria but the viability (or recovery rate) of cells after sorting has not been clarified in the current research progress. In this work, to keep the cells alive as much as possible, we propose a three-layer LIFT system (top layer: 25-nm aluminum film; second layer: 3 μm agar media; third layer: liquid containing bacterial) for the isolation and cultivation of single Gram-negative ( E. coli ), Gram-positive ( Lactobacillus rhamnosus GG, LGG), and eukaryotic microorganisms ( Saccharomyces cerevisiae ). The experiment results showed that the average survival rates for ejected pure single cells were 63% for Saccharomyces cerevisiae , 22% for E. coli DH5α, and 74% for LGG. In addition, we successfully isolated and cultured the GFP expressing E. coli JM109 from the mixture containing complex communities of soil bacteria by fluorescence signal. The average survival rate of E. coli JM109 was demonstrated to be 25.3%. In this study, the isolated and cultured single colonies were further confirmed by colony PCR and sequencing. Such precise sorting and cultivation technique of live single microbial cells could be coupled with other microscopic approaches to isolate single microorganisms with specific functions, revealing their roles in the natural community. Importance We developed a laser induced forward transfer (LIFT) technology to accurately isolate single live microbial cells. The cultivation recovery rates of the ejected single cells were 63% for Saccharomyces cerevisiae , 22% for E. coli DH5α, and 74% for Lactobacillus rhamnosus GG (LGG). Coupled LIFT with fluorescent microscope, we demonstrated that single cells of GFP expressing E. coli JM109 were sorted according to fluorescence signal from a complex community of soil bacteria, and subsequently cultured with 25% cultivation recovery rate. This single cell live sorting technology could isolate single microbes with specific functions, revealing their roles in the natural community.

Imaging Flow Cytometry to Study Biofilm-Associated Microbial Aggregates

The aim of the research was to design an advanced analytical tool for the precise characterization of microbial aggregates from biofilms formed on food-processing surfaces. The approach combined imaging flow cytometry with a machine learning-based interpretation protocol. Biofilm samples were collected from three diagnostic points of the food-processing lines at two independent time points. The samples were investigated for the complexity of microbial aggregates and cellular metabolic activity. Thus, aggregates and singlets of biofilm-associated microbes were simultaneously examined for the percentages of active, mid-active, and nonactive (dead) cells to evaluate the physiology of the microbial cells forming the biofilm structures. The tested diagnostic points demonstrated significant differences in the complexity of microbial aggregates. The significant percentages of the bacterial aggregates were associated with the dominance of active microbial cells, e.g., 75.3% revealed for a mushroom crate. This confirmed the protective role of cellular aggregates for the survival of active microbial cells. Moreover, the approach enabled discriminating small and large aggregates of microbial cells. The developed tool provided more detailed characteristics of bacterial aggregates within a biofilm structure combined with high-throughput screening potential. The designed methodology showed the prospect of facilitating the detection of invasive biofilm forms in the food industry environment.

Biofilms from Pathogenic Bacteria in Food Processing Environments: Formation and Preventive Disinfection Procedures

The ability of some pathogenic bacterial species to form biofilms on surfaces of equipment and utensils is of great concern to the food industry since they represent a continuous source of contamination in food processing environments. In this review, the factors involved in the formation of microbial biofilms are highlighted, along with a discussion on the preventive disinfection procedures recommended to avoid the attachment of microbial cells on surfaces of equipment and utensils in food processing areas. Relevant articles published in the last 10 years (2012-present) were selected in PubMed, Science Direct, and Google Scholar. Methods for assessing the adhesion and biofilm formation ability of strains isolated from surfaces in the food industry environment are also presented.