Appl. Microbiol., Volume 1, Issue 2 (September 2021) – 14 articles

The bioprocessing strategy is an effective approach to improve bioavailability and stability of bioactive compounds for designing functional foods and ingredients. In this study, food barley was bio-transformed to improve functional bioactives by sprouting, coupled with beneficial lactic acid bacteria (LAB)-based fermentation. Dairy Kefir culture with mixed beneficial LAB strains was targeted to ferment aqueous slurries of sprouted hulless food barley flour (unpigmented, purple, and black barley) for 72 h, and modulation of phenolic-linked antioxidant and anti-hyperglycemic functionalities were evaluated using in vitro assay models. The biochemical parameters analyzed were total soluble phenolic (TSP) content, profile of phenolic compounds, total antioxidant activity, and anti-hyperglycemic property-relevant α-amylase and α-glucosidase enzyme inhibitory activities. Furthermore, human gut health benefits of relevant properties of fermented slurries of barley flour were also evaluated based on growth of Kefir culture and subsequent determination of anti-bacterial potential against pathogenic human ulcer causing bacteria Helicobacter pylori. Kefir culture-mediated fermentation of 48-h sprouted barley flours improved the TSP content and associated antioxidant and anti-hyperglycemic functionalities. Additionally, anti-bacterial potential against H. pylori and sustaining active growth of viable LAB cells above the minimum level required for probiotic activity were also observed in fermented food barley flour slurries. Full article

Due to the importance of microbes in soil health and crop production, manipulation of microbiomes provides a new strategy for improving crop growth and agricultural ecosystems. Current understanding is limited regarding the responses of soil and crop endophytic microbiomes to field management and microbiome programming. In this study, we investigated soil and tea root bacterial communities under conventional and organic cropping systems using 16S rRNA gene sequencing. A significant difference in soil and root bacterial community structure was observed under different field managements, leading to 43% and 35% variance, respectively. We also identified field management-sensitive species both in soils and tea roots that have great potential as bioindicators for bacterial microbiome manipulation. Moreover, through functional profile predictions of microbiomes, xenobiotics degradation in soil bacterial communities is enriched in organic farms, suggesting that biodegradation capabilities are enhanced under organic cropping systems. Our results demonstrate the effects of field management on both soil and tea root bacterial microbiomes and provide new insights into the reprogramming of microbial structures. Full article

Brettanomyces bruxellensis is a wine spoilage concern in wineries around the world. In order to maintain wine quality during storage and ageing, it is imperative to control and monitor this yeast. Being a fastidious slow growing yeast, which requires 5 to 14 days of incubation for visible growth in agar plates, it is difficult to detect growth (colonies) by conventional agar plate count method. Yeast enumeration by impedance was investigated because previous research using other microorganisms has shown that it is potentially faster than plate counting. The relationship between plate counting and impedance detection times was investigated for Brettanomyces inoculated in red wine samples. A linear relationship between log plate count concentrations and impedance detection times was found. Incubation time was reduced from 120 h down to 0.9 and 57.7 h for samples with 6.7 × 10⁷ and 1.8 × 10² cfu/mL, respectively, using the ‘indirect’ impedance method. The ‘direct’ method also reduced the incubation times to 9.5 and 81.9 h, for the same concentrations. The ‘indirect’ impedance method has the potential to be used by the wine industry to control and monitor the Brettanomyces numbers in wines. Full article

Soil contamination with petroleum hydrocarbons (PHCs) has become a global concern and has resulted from the intensification of industrial activities. This has created a serious environmental issue; therefore, there is a need to find solutions, including application of efficient remediation technologies or improvement of current techniques. Rhizoremediation is a green technology that has received global attention as a cost-effective and possibly efficient remediation technique for PHC-polluted soil. Rhizoremediation refers to the use of plants and their associated microbiota to clean up contaminated soils, where plant roots stimulate soil microbes to mineralize organic contaminants to H₂O and CO₂. However, this multipartite interaction is complicated because many biotic and abiotic factors can influence microbial processes in the soil, making the efficiency of rhizoremediation unpredictable. This review reports the current knowledge of rhizoremediation approaches that can accelerate the remediation of PHC-contaminated soil. Recent approaches discussed in this review include (1) selecting plants with desired characteristics suitable for rhizoremediation; (2) exploiting and manipulating the plant microbiome by using inoculants containing plant growth-promoting rhizobacteria (PGPR) or hydrocarbon-degrading microbes, or a combination of both types of organisms; (3) enhancing the understanding of how the host–plant assembles a beneficial microbiome, and how it functions, under pollutant stress. A better understanding of plant–microbiome interactions could lead to successful use of rhizoremediation for PHC-contaminated soil in the future. Full article

The anaerobic fermentation of glucose and fructose was performed by Actinobacillus succinogenes 130Z in batch mode using three different volume of bioreactors (0.25, 1 and 3 L). The strategy used was the addition of MgCO₃ and fumaric acid (FA) as mineral carbon and the precursor of succinic acid, respectively, in the culture media. Kinetics and yields of succinic acid (SA) production in the presence of sugars in a relevant synthetic medium were investigated. Work on the bench scale (3 L) showed the best results when compared to the small anaerobic reactor’s succinic acid yield and productivity after 96 h of fermentation. For an equal mixture of glucose and fructose used as substrate at 0.4 mol L⁻¹ with the addition of FA as enhancer and under proven optimal conditions (pH 6.8, T = 37 °C, anaerobic condition and 1% v/v of biomass), about 0.5 mol L⁻¹ of SA was obtained, while the theoretical production of succinic acid was 0.74 mol L⁻¹. This concentration corresponded to an experimental yield of 0.88 (mol-C SA/mol-C sugars consumed anaerobically) and a volumetric productivity of 0.48 g-SA L⁻¹ h⁻¹. The succinic acid yield and concentration obtained were significant and in the order of those reported in the literature. Full article

Background: the aim of this study was to improve the overall microbiological quality of five different sprouts (alfalfa, kale, lentil, sunflower, radish) using newly isolated bacteriophages. Method: in this study we had isolated from sewage 18 bacteriophages targeting bacteria dominant in sprouts. Five selected bacteriophage strains were photographed using a transmission electron microscope (TEM), and we analyzed the rate of attachment, resistance to chloroform, the burst size, and the latency period. Two methods of application of the phage cocktail were investigated: spraying, and an absorption pad. Results: the spraying method was significantly more efficient, and the maximum reduction effect after 48 h of incubation was 1.5 log CFU/g. Using pads soaked with phage lysate reduced the total number of bacteria to only about 0.27–0.79 log CFU/g. Conclusion: the reduction of bacteria levels in sprouts depended on the method of phage application. The blind strategy for searching phage targeting bacteria dominant in sprouts can be useful and economically beneficial as a starting point for further investigation in phage cocktail application for improving the overall microbiological quality of food. The main result of our research is to improve the overall quality of kale and radish sprouts by spraying them with a phage cocktail. Full article

Beneficial lactic acid bacteria (LAB)-based fermentation is an effective bioprocessing approach to improve human-health-targeted functional benefits of plant-based food substrates, such as cereal grains. Previously, we observed high phenolic bioactive-linked antioxidant and anti-hyperglycemic properties in whole grain Emmer (hulled). In this study, beneficial LAB (Lactiplantibacillus plantarum) was recruited to ferment (0–72 h) aqueous extracts (0.4 g/mL concentration) of previously optimized hulled Emmer wheat and conventional red spring wheat cv. Barlow. The fermented and unfermented (control) wheat extracts were analyzed for phenolic content, phenolic profile, antioxidant activity, and antihyperglycemic properties (α-amylase and α-glucosidase enzyme inhibitory activity) using in vitro assay models. Additionally, antimicrobial activity against pathogenic bacteria Helicobacter pylori, and potential prebiotic activity supporting the growth of beneficial Bifidobacterium longum were also investigated. Improvement in antioxidant activity and antihyperglycemic functional benefits were observed, while soluble phenolic content remained high after 72 h fermentation. Antimicrobial activity against H. pylori was also observed in 48 and 72 h fermented wheat extracts. This study provides an insight into the efficacy of LAB-based fermentation as a safe bioprocessing tool to design health-targeted functional foods and ingredients from underutilized whole grains like Emmer for targeting type 2 diabetes dietary benefits. Full article

Microbial attachment to surfaces is ubiquitous in nature. Most species of bacteria attach and adhere to surfaces via special appendages such as pili and fimbriae, the roles of which have been extensively studied. Here, we report an experiment on pilus-less mutants of Caulobacter crescentus weakly attached to polyethylene surface. We find that some individual cells transiently but repeatedly adhere to the surface in a stick-slip fashion in the presence of an electric field parallel to the surface. These bacteria move significantly slower than the unattached ones in the same field of view undergoing electrophoretic motion. We refer this behavior of repeated and transient attachment as “quasi-attachment”. The speed of the quasi-attached bacteria exhibits large variation, frequently dropping close to zero for short intervals of time. We propose a polymeric tethering model to account for the experimental findings. This study sheds light on bacteria–surface interaction, which is significant in broader contexts such as infection and environmental control. Full article

Recently, there has been a resurgence of interest in continuous bioprocessing as a cost-optimised production strategy, driven by a rising global requirement for recombinant proteins used as biological drugs. This strategy could provide several benefits over traditional batch processing, including smaller bioreactors, smaller facilities, and overall reduced plant footprints and investment costs. Continuous processes may also offer improved product quality and minimise heterogeneity, both in the culture and in the product. In this paper, a model protein, green fluorescent protein (GFP) mut3*, was used to test the recombinant protein expression in an Escherichia coli strain with industrial relevance grown in chemostat. An important factor in enabling stable productivity in continuous cultures is the carbon source. We have studied the viability and heterogeneity of the chemostat cultures using a chemically defined medium based on glucose or glycerol as the single carbon source. As a by-product of biodiesel production, glycerol is expected to become a sustainable alternative substrate to glucose. We have found that although glycerol gives a higher cell density, it also generates higher heterogeneity in the culture and a less stable recombinant protein production. We suggest that manipulating the balance between different subpopulations to increase the proportion of productive cells may be a possible solution for making glycerol a successful alternative to glucose. Full article

DNA double-strand breaks (DSBs) are the most deleterious form of DNA damage and are repaired through non-homologous end-joining (NHEJ) or homologous recombination (HR). Repair initiation, regulation and communication with signaling pathways require several histone-modifying and chromatin-remodeling complexes. In budding yeast, this involves three primary complexes: INO80-C, which is primarily associated with HR, SWR1-C, which promotes NHEJ, and RSC-C, which is involved in both pathways as well as the general DNA damage response. Here we identify ARP6 as a factor involved in DSB repair through an RSC-C-related pathway. The loss of ARP6 significantly reduces the NHEJ repair efficiency of linearized plasmids with cohesive ends, impairs the repair of chromosomal breaks, and sensitizes cells to DNA-damaging agents. Genetic interaction analysis indicates that ARP6, MRE11 and RSC-C function within the same pathway, and the overexpression of ARP6 rescues rsc2∆ and mre11∆ sensitivity to DNA-damaging agents. Double mutants of ARP6, and members of the INO80 and SWR1 complexes, cause a significant reduction in repair efficiency, suggesting that ARP6 functions independently of SWR1-C and INO80-C. These findings support a novel role for ARP6 in DSB repair that is independent of the SWR1 chromatin remodeling complex, through an apparent RSC-C and MRE11-associated DNA repair pathway. Full article

Sea urchins, in many instances, are collected from the wild, maintained in the laboratory aquaculture environment, and used as model animals for various scientific investigations. It has been increasingly evident that diet-driven dysbiosis of the gut microbiome could affect animal health and physiology, thereby impacting the outcome of the scientific studies. In this study, we compared the gut microbiome between naturally occurring (ENV) and formulated diet-fed laboratory aquaculture (LAB) sea urchin Lytechinus variegatus by amplicon sequencing of the V4 region of the 16S rRNA gene and bioinformatics tools. Overall, the ENV gut digesta had higher taxa richness with an abundance of Propionigenium, Photobacterium, Roseimarinus, and Flavobacteriales. In contrast, the LAB group revealed fewer taxa richness, but noticeable abundances of Arcobacter, Agarivorans, and Shewanella. However, Campylobacteraceae, primarily represented by Arcobacter spp., was commonly associated with the gut tissues of both ENV and LAB groups whereas the gut digesta had taxa from Gammaproteobacteria, particularly Vibrio spp. Similarly, the co-occurrence network displayed taxonomic organizations interconnected by Arcobacter and Vibrio as being the key taxa in gut tissues and gut digesta, respectively. Predicted functional analysis of the gut tissues microbiota of both ENV and LAB groups showed a higher trend in energy-related metabolisms, whereas amino acids, carbohydrate, and lipid metabolisms heightened in the gut digesta. This study provides an outlook of the laboratory-formulated diet-fed aquaculture L. variegatus gut microbiome and predicted metabolic profile as compared to the naturally occurring animals, which should be taken into consideration for consistency, reproducibility, and translatability of scientific studies. Full article

High-resolution X-ray computer tomography (microCT) is a well-established technique to analyse three-dimensional microstructures in 3D non-destructive imaging. The non-destructive three-dimensional analysis of lichens is interesting for many reasons. The examination of hidden structural characteristics can, e.g., provide information on internal structural features (form and distribution of fungal-supporting tissue/hypha), gas-filled spaces within the thallus (important for gas exchange and, thus, physiological processes), or yield information on the symbiont composition within the lichen, e.g., the localisation and amount of additional cyanobacteria in cephalodia. Here, we present the possibilities and current limitations for applying conventional laboratory-based high-resolution X-ray computer tomography to analyse lichens. MicroCT allows the virtual 3D reconstruction of a sample from 2D X-ray projections and is helpful for the non-destructive analysis of structural characters or the symbiont composition of lichens. By means of a quantitative 3D image analysis, the volume of internal cephalodia is determined for Lobaria pulmonaria and the external cephalodia of Peltigera leucophlebia. Nevertheless, the need for higher-resolution tomography for more detailed studies is emphasised. Particular challenges are the large sizes of datasets to be analysed and the high variability of the lichen microstructures. Full article

Viral infections are known to modulate the upper respiratory tract microbiome, but few studies have addressed differences in the nasopharyngeal microbiome following SARS-CoV-2 infection. Using nasopharyngeal swab medical waste samples from 79 confirmed SARS-CoV-2 positive and 20 SARS-CoV-2 negative patients, we assessed microbiome composition with metagenomic sequencing. COVID-19 status and breathing assistive device use was associated with differences in beta diversity, principal component analyses, community composition and abundance of several species. Serratia more frequently appeared in COVID-19 patient samples compared to negative patient samples, and Serratia, Streptococcus, Enterobacter, Veillonella, Prevotella, and Rothia appeared more frequently in samples of those who used breathing assistive devices. Smoking and age were associated with differences in alpha diversity. Cross-sectional differences in the microbiome were apparent with SARS-CoV-2 infection, but longitudinal studies are needed to understand the dynamics of viral and breathing treatment modulation of microbes. Full article

Solanum tuberosum, commonly known as potato, is the most important non-cereal crop in the world. However, its cultivation is prone to disease and other issues. In recent years, a newfound interest in the soil microbiome and the potential benefits it may convey has led researchers to study plant–microbe interactions in great detail and has led to the identification of putative beneficial microbial taxa. In this survey, we examined fungal and bacterial diversity using high-throughput sequencing in soils under a potato crop in southeastern Wyoming, USA. Our results show decreased microbial diversity in the rhizosphere, with increases in the abundances of arbuscular mycorrhizal fungi as well as pathogenic microbes. We show coarse taxonomic differences in microbial assemblages when comparing the bulk and rhizosphere soils for bacteria but not for fungi, suggesting that the two kingdoms respond differently to the selective pressures of the rhizosphere. Using cooccurrence network analysis, we identify microbes that may serve as keystone taxa and provide benefits to their host plants through competitive exclusion of detrimental pathogenic taxa and increased nutrient availability. Our results provide additional information on the structure and complexity of the potato rhizosphere microbiome and highlight candidate taxa for microbial isolation and inoculation. Full article