Effect of pH-shifting on the water holding capacity and gelation properties of mung bean protein isolate.

In this study, alkaline pH-shifting modified the globular structure of mung bean protein isolate (MBPI) to form flexible and stretched structures. In contrast, acidic pH-shifting increased the rigidity of MBPI. The increased flexibility (at the level of the secondary structure) and newly exposed intermolecular amino acid groups induced by alkaline pH-shifting improved the water holding capacity and gelation properties of proteins. Specifically, MBPI treated at pH 12 (MP12) showed the most flexible structure and highest water holding capacity and gel formation properties (least gelation concentration). The water-holding capacity of native MBPI increased from 1.56 g/g to 4.81 g/g, and its least gelation concentration decreased from 22 % to 15 % by pH-shifting at pH 12. Furthermore, MP12 formed stronger and more elastic heat-induced gels than native MBPI. We identified significant differences in the structural properties and water holding capacity, and gelation properties of acidic and alkaline pH-shifted MBPI and investigated the gelation properties of MP12 including rheological and morphological analyses. Our findings can facilitate the use of mung beans as a protein source in a wide range of food applications, including plant-based and processed meats.

Impact of propiconazole fungicide on soil microbiome (bacterial and fungal) diversity, functional profile, and associated dehydrogenase activity.

Pesticides, protect crops but can harm the environment and human health when used without caution. This study evaluated the impact of propiconazole, a fungicide that acts on fungal cell membranes, on soil microbiome abundance, diversity, and functional profile, as well as soil dehydrogenase activity (DHA). The study conducted microcosm experiments using soil samples treated with propiconazole and employed next-generation sequencing (MiSeq) and chromatographic approaches (GC-MS/MS) to analyze the shift in microbial communities and propiconazole level, respectively. The results showed that propiconazole significantly altered the distribution of microbial communities, with notable changes in the abundance of various bacterial and fungal taxa. Among soil bacterial communities, the relative abundance of Proteobacteria and Planctomycetota increased, while that of Acidobacteria decreased after propiconazole treatment. In the fungal communities, propiconazole increased the abundance of Ascomycota and Basidiomycota in the treated soil, while that of Mortierellomycota was reduced. Fungicide application further triggered a significant decrease in DHA over time. Analysis of the functional profile of bacterial communities showed that propiconazole significantly affected bacterial cellular and metabolic pathways. The carbon degradation pathway was upregulated, indicating the microbial detoxification of the contaminant in the treated soil. Our findings suggest that propiconazole application has a discernible impact on soil microbial communities, which could have long-term consequences for soil health, quality, and function.

Deciphering key factors in pathogen-suppressive microbiome assembly in the rhizosphere.

In a plant-microbe symbiosis, the host plant plays a key role in promoting the association of beneficial microbes and maintaining microbiome homeostasis through microbe-associated molecular patterns (MAMPs). The associated microbes provide an additional layer of protection for plant immunity and help in nutrient acquisition. Despite identical MAMPs in pathogens and commensals, the plant distinguishes between them and promotes the enrichment of beneficial ones while defending against the pathogens. The rhizosphere is a narrow zone of soil surrounding living plant roots. Hence, various biotic and abiotic factors are involved in shaping the rhizosphere microbiome responsible for pathogen suppression. Efforts have been devoted to modifying the composition and structure of the rhizosphere microbiome. Nevertheless, systemic manipulation of the rhizosphere microbiome has been challenging, and predicting the resultant microbiome structure after an introduced change is difficult. This is due to the involvement of various factors that determine microbiome assembly and result in an increased complexity of microbial networks. Thus, a comprehensive analysis of critical factors that influence microbiome assembly in the rhizosphere will enable scientists to design intervention techniques to reshape the rhizosphere microbiome structure and functions systematically. In this review, we give highlights on fundamental concepts in soil suppressiveness and concisely explore studies on how plants monitor microbiome assembly and homeostasis. We then emphasize key factors that govern pathogen-suppressive microbiome assembly. We discuss how pathogen infection enhances plant immunity by employing a cry-for-help strategy and examine how domestication wipes out defensive genes in plants experiencing domestication syndrome. Additionally, we provide insights into how nutrient availability and pH determine pathogen suppression in the rhizosphere. We finally highlight up-to-date endeavors in rhizosphere microbiome manipulation to gain valuable insights into potential strategies by which microbiome structure could be reshaped to promote pathogen-suppressive soil development.

Spinach (Spinacia oleracea) as green manure modifies the soil nutrients and microbiota structure for enhanced pepper productivity.

Spinach has been suggested as a potential rotation crop for increasing crop yield by enhancing beneficial fungal microbes in continuous monocropping. However, no research on the use of spinach as a green manure has been reported. Thus, we tested the effects of spinach and Korean mustard cultivars (green and red mustards) (10 g pot -1) as green manure on soil chemical properties, pepper productivity, and soil microbiome of long-year pepper-monocropped soil. Spinach improved the soil nutrition (e.g., pH, SOM, TN, NH4+, and K), weed suppression, and pepper growth. Spinach had by far the highest fruit yield, over 100% pepper fruit yield increment over the mustard green manures and control. Our study showed that the major influencing factors to cause a shift in both bacterial and fungal community assemblies were soil pH, TC TN, and K. Following green manure amendment Bacillota, especially Clostridium, Bacillus and Sedimentibacter, were enriched, whereas Chloroflexi and Acidobacteriota were reduced. In addition, spinach highly reduced the abundance of Leotiomycetes and Fusarium but enriched Papiliotrema. FAPROTAX and FUNGuild analysis revealed that predicted functional profiles of bacterial and fungal communities in spinach-amended soil were changed. Spinach-treated soil was differentially abundant in function related to hydrocarbon degradation and functional guilds of symbiotrophs and ectomycorrhizal. This study contributes significantly to our understanding of how the soil fertility and soil microbiome alteration via spinach green manure application as a pre-plant soil treatment might help alleviate continuous cropping obstacles.

Soil amendment with cow dung modifies the soil nutrition and microbiota to reduce the ginseng replanting problem.

Ginseng is a profitable crop worldwide; however, the ginseng replanting problem (GRP) is a major threat to its production. Soil amendment is a non-chemical method that is gaining popularity for alleviating continuous cropping obstacles, such as GRP. However, the impact of soil amendment with either cow dung or canola on GRP reduction and the associated soil microbiota remains unclear. In the present study, we evaluated the effect of soil amendment with cow dung, canola seed powder, and without amendment (control), on the survival of ginseng seedling transplants, the soil bacterial and fungal communities, and their associated metabolic functions. The results showed that cow dung increased ginseng seedling survival rate by 100 percent and had a remarkable positive effect on ginseng plant growth compared to control, whereas canola did not. Cow dung improved soil nutritional status in terms of pH, electrical conductivity, NO 3 - , total carbon, total phosphorus, and available phosphorus. The amplicon sequencing results using Illumina MiSeq showed that canola had the strongest negative effect in reducing soil bacterial and fungal diversity. On the other hand, cow dung stimulated beneficial soil microbes, including Bacillus, Rhodanobacter, Streptomyces, and Chaetomium, while suppressing Acidobacteriota. Community-level physiological profiling analysis using Biolog Ecoplates containing 31 different carbon sources showed that cow dung soil had a different metabolic activity with higher utilization rates of carbohydrates and polymer carbon sources, mainly Tween 40 and beta-methyl-d-glucoside. These carbon sources were most highly associated with Bacillota. Furthermore, predicted ecological function analyses of bacterial and fungal communities showed that cow dung had a higher predicted function of fermentation and fewer functions related to plant pathogens and fungal parasites, signifying its potential to enhance soil suppressiveness. Co-occurrence network analysis based on random matrix theory (RMT) revealed that cow dung transformed the soil microbial network into a highly connected and complex network. This study is the first to report the alleviation of GRP using cow dung as a soil amendment, and the study contributes significantly to our understanding of how the soil microbiota and metabolic alterations via cow dung can aid in GRP alleviation.

Comparative analysis of scalp and gut microbiome in androgenetic alopecia: A Korean cross-sectional study.

Androgenetic alopecia (AGA) is a non-scarring and progressive form of hair loss occurring in both men and women. Although genetic predisposition and sex steroid hormones are the main causes, many factors remain unknown, and various extrinsic factors can negatively affect the lifespan of hair. We investigated skin-gut axis microorganisms as potential exogenous factors causing AGA, through comparative analyses of the scalp and gut microbiome in individuals with and without AGA in a Korean cohort. Using 16S rRNA gene sequencing, we characterized the scalp and gut microbiomes of 141 individuals divided into groups by sex and presence of AGA. Alpha diversity indices in the scalp microbiome were generally higher in individuals with AGA than in healthy controls. These indices showed a strong negative correlation with scalp-inhabitant bacteria (Cutibacterium and Staphylococcus), indicating that the appearance of non-inhabitant bacteria increases as hair loss progresses. No significant differences in diversity were observed between the gut microbiomes. However, bacterial functional differences, such as bile acid synthesis and bacterial invasion of epithelial cells, which are related to intestinal homeostasis, were observed. The networks of the scalp and gut microbiome were more complex and denser with higher values of the network topology statistic coefficient values (i.e., transitivity, density, and degree centrality) and more unique associations in individuals with AGA than in healthy controls. Our findings reveal a link between skin-gut microorganisms and AGA, indicating the former's potential involvement in the latter's development. Additionally, these results provide evidence for the development of cosmetics and therapeutics using microorganisms and metabolites involved in AGA.

Complete Genome Sequence of Bacillus amyloliquefaciens KNU-28 Isolated from Peach Leaves (Prunus Persica [L.] Batsch).

This study presented the complete genome sequence of B. amyloliquefaciens KNU-28 isolated from the leaves of the peach (Prunus persica [L.] Batsch). The genome of this strain comprised one chromosome with 4,238,926 bp and 45.9% of GC content.

Development of mixed starter culture for the fermentation of Ethiopian honey wine, Tej.

Ethiopian honey wine is one of the country's most popular spontaneously fermented traditional alcoholic beverages. However, the final product of this natural fermentation system is frequently of poor and inconsistent quality. Furthermore, it makes the process difficult to predict, control, and correct. Thus, the main aim of this study was to develop a direct fermentation system for Ethiopian honey wine, Tej. After isolating fermentative microbial strains from Tej samples, they were subjected to intensive screening to fit to its purpose. Later, phenotypic and genotypic characterization, and inoculation of isolates to honey-must were performed sequentially. Finally, microbial interaction and physicochemical analysis, including volatile compounds profiling, were done for the inoculated samples. The identified isolates were strains of Saccharomycetaceae and Lactobacillaceae families. These strains showed a good ability to tolerate osmotic stress and a lower pH environment. Tej sample produced by mixed culture inoculation of Saccharomyces and Lactobacillus species showed similar physicochemical, volatile compounds, and sensory attributes values with that of the control sample. Thus, a mixture of Saccharomyces and Lactobacillus strains could be used as a starter culture to produce Ethiopian honey, Tej, without scarifying of its major quality attributes.

Bioengineered Lab-Grown Meat-like Constructs through 3D Bioprinting of Antioxidative Protein Hydrolysates.

Lab-grown bovine meat analogues are emerging alternatives to animal sacrifices for cultured meat production. The most challenging aspect of the production process is the rapid proliferation of cells and establishment of the desired 3D structure for mass production. In this study, we developed a direct ink writing-based 3D-bioprinted meat culture platform composed of 6% (w/v) alginate and 4% (w/v) gelatin (Alg/Gel)-based hydrogel scaffolds supplemented with naturally derived protein hydrolysates (PHs; 10%) from highly nutritive plants (soybean, pigeon pea, and wheat), and some selected edible insects (beetles, crickets, and mealworms) on in vitro proliferation of bovine myosatellite cells (bMSCs) extracted from fresh meat samples. The developed bioink exhibited excellent shear-thinning behavior (n < 1) and mechanical stability during 3D bioprinting. Commercial proteases (Alcalase, Neutrase, and Flavourzyme) were used for protein hydrolysis. The resulting hydrolysates exhibited lower-molecular-weight bands (12-50 kDa) than those of crude isolates (55-160 kDa), as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The degree of hydrolysis was higher in the presence of Alcalase for both plant (34%) and insect (62%) PHs than other enzymes. The 3D-printed hydrogel scaffolds displayed excellent bioactivity and stability after 7 days of incubation. The developed prototype structure (pepperoni meat, 20 × 20 × 5 mm) provided a highly stable, nutritious, and mechanically strong structure that supported the rapid proliferation of myoblasts in a low-serum environment during the entire culture period. The 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay enhanced the free radical reduction of Alcalase- and Neutrase-treated PHs. Furthermore, the bioprinted bMSCs displayed early myogenesis (desmin and Pax7) in the presence of PHs, suggesting its role in bMSC differentiation. In conclusion, we developed a 3D bioprinted and bioactive meat culture platform using Alg/Gel/PHs as a printable and edible component for the mass production of cultured meat.

Unraveling the phase behavior of cricket protein isolate and alginate in aqueous solution.

The associative phase behavior of cricket protein isolate (CPI) and sodium alginate (AL) in aqueous solutions was explored using turbidimetry, methylene blue spectroscopy, zeta potentiometry, dynamic light scattering, and confocal microscopy as a function of pH, biopolymer ratio, total biopolymer concentration (CT), and ionic strength. When both biopolymers had net-negative charges, soluble complexes formed between pH 6.0 and 8.0, however when both biopolymers had opposing net charges, insoluble complexes formed as complex coacervates below pH 5.5, defined as pHφ1, followed by precipitates below another critical pH 3.0 (pHp). Increasing the CPI:AL weight ratio or CT facilitated complex formation, and the addition of salts (NaCl/KCl) had a salt-enhancement and salt-reduction impact at low and high salt concentrations, respectively. Ionic interactions between oppositely charged CPI and AL were mainly responsible for the formation of their insoluble complexes, while hydrogen bonding and hydrophobic interactions also played significant roles.

Microbiome dataset of spontaneously fermented Ethiopian honey wine, Tej.

This dataset contains raw and analyzed microbial data for the samples of spontaneously fermented Ethiopian honey wine, Tej, collected from three locations of Ethiopia. It was generated using culture independent amplicon sequencing technique. To gain a better understanding of microbial community variance and similarity across Tej samples from the same and different locations, the raw sequenced data obtained from the Illumina Miseq sequencer was subjected to a bioinformatics analysis. Lower diversity and richness of both bacterial and fungal communities were observed for all of the Tej samples. Besides, samples collected from Debre Markos area showed a significant discriminating tax for both bacterial and fungal communities. In nutshell, this amplicon sequencing dataset provides a useful collection of data for modernizing this spontaneous fermentation into a directed inoculated fermentation. Detail discussion on microbiome of Tej samples is given in [1].

Physicochemical properties, antioxidant activities and microbial communities of Ethiopian honey wine, Tej.

Ethiopian honey wine, Tej, is spontaneously fermented traditional alcoholic beverage, usually made from honey and "gesho" (Rhamnus prinoides). Till now, limited amount of information is available on the characterization of Tej. Thus, the aim of this paper is to reveal the microbiological diversity and physicochemical properties of Tej samples collected from different areas of Ethiopia. High-throughput sequencing, electrochemical and chromatographic techniques, and spectrophotometric methods were used to achieve these objectives. Although there was a statistical difference in the exact values of physicochemical properties between the collected Tej samples, the pH and titratable acidity values of the samples ranged from 2.8 to 3.8 and from 1.81 to 8.65 g/L, respectively. Similarly, the alcohol and sugar contents of the samples were in the range of 6.36-11.34 g/100 mL and from 0.37 to 31.6 g/L, respectively. Moreover, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 3-ethylbenzothiazoline-6-sulfonic acid diammonium salt (ABTS) values of the samples were in the range of 37.9-81.0% and 27.4-73.1%, respectively. Furthermore, microbial community structure was predominated by a few fermentative microorganisms. Specifically, the bacterial community structure was dominated by the genera of Lactobacillus (53.15%) and Zymomonas (38.41%). Whereas, the fungal community structure was exclusively dominated by genus of Saccharomyces (99.66%). Additionally, Lactobacillus, Zymomonas and Saccharomyces were the detected core microbiome for the collected Tej samples. Both bacterial and fungal communities had shown no statistically significant differences in alpha diversity analysis based on the area of sample collection. However, the bacterial communities had a statically significant difference in Unweighted Unifrac beta diversity analysis. Generally, the observed shared physicochemical characteristic features and the dominance by certain group of microorganisms might be seen as a boon for the development of direct fermentation system to this traditional alcoholic beverage.

Alterations and Prediction of Functional Profiles of Gut Microbiota After Fecal Microbiota Transplantation for Iranian Recurrent Clostridioides difficile Infection with Underlying Inflammatory Bowel Disease: A Pilot Study.

BACKGROUND AND PURPOSE: Fecal microbiota transplantation (FMT) has emerged for the therapeutic treatment of recurrent Clostridioides difficile infection (rCDI) with concurrent inflammatory bowel disease (IBD). As the first Iranian population cohort, we examined how gut microbiota and their functional profiles change in Iranian rCDI patients with underlying IBD before and after FMT. PATIENTS AND METHODS: FMT was performed to eight IBD patients via colonoscopy. Profiles of gut microbiota from donors and recipients were investigated using 16S rRNA gene sequence analysis. RESULTS: Patients experienced no IBD flare-ups or other adverse effects, and all recovered to full health. Moreover, all rCDI patients lacked the Bacteroidetes present in donor samples. After FMT, the proportion of Bacteroidetes increased until a normal range was achieved. More specifically, the relative abundance of Prevotella was found to increase significantly following FMT. Prevotella was also found to correlate negatively with inflammation metrics, suggesting that Prevotella may be a key factor for resolving CDI and IBD. Gut microbiota diversity was found to increase following FMT, while dysbiosis decreased. However, the similarity of microbial communities of host and recipients did not increase, and wide variation in the extent of donor stool engraftment indicated that the gut bacterial communities of recipients do not shift towards those of donors. CONCLUSION: FMT leads to significant alterations of the community structure of gut bacteria in rCDI patients with IBD. The change in relative abundance of Proteobacteria and bacterial diversity indicated that FMT promotes recovery from intestinal permeability and inflammation in rCDI patients. Moreover, strong negative correlation between Prevotella and inflammation index, and decreased dysbiosis index advocate that the improvement of CDI is possibly due to gut microbiome alteration. Collectively, our findings show that FMT would be a promising therapy to help reprogram the gut microbiome of Iranian rCDI patients with IBD.

Bacterial Profiles of Brain in Downer Cattle with Unknown Etiology.

Downer cow can be caused by muscular paralysis, neurological damage, metabolic disorder, and/or the complication of microbial infection. However, downer cow with unknown etiology is issued because of the non-detection of its bacterial etiological agent. In this study, differences in the bacterial community in brain tissues between downer cattle with unknown etiology and healthy slaughtered cattle are investigated. Bacterial diversity and representative genera between downer and normal cattle were significantly different (p < 0.05). There are significant differences in representative genera of downer and normal cattle, especially the significance, fold change, and area under the receiver operating characteristic curve score (p < 0.05). Furthermore, the prediction of functional genes in brain microbiota between the downer and normal cattle revealed differences in the cluster of orthologous gene categories, such as lipid transport and metabolism, secondary metabolite biosynthesis, and signal transduction (p < 0.05). This study revealed a significant difference in microbiota between the downer and normal cattle. Thus, we demonstrate that representative genera from downer cattle through 16S rRNA gene amplicon sequencing and microbiota analysis have the potential as candidates for bacterial etiological agents for downer cow.

Effect of Three Defatting Solvents on the Techno-Functional Properties of an Edible Insect (Gryllus bimaculatus) Protein Concentrate.

Edible insects have received global attention as an alternative protein-rich food. However, their structural characteristics make them difficult to digest. To overcome this obstacle, we assessed the techno-functional properties of three protein concentrates from the cricket Gryllus bimaculatus. Freeze-dried G. bimaculatus powder was defatted using ethanol, hexene, or acetone as solvents, and the techno-functional properties (protein solubility, water and oil holding capacity, foaming properties, emulsion capacity, and gel formation) of the protein concentrates were determined. Freeze-dried G. bimaculatus powder comprised approximately 17.3% crude fat and 51.3% crude protein based on dry weight. Ethanol was the most effective solvent for reducing the fat content (from 17.30% to 0.73%) and increasing the protein content (from 51.3% to 62.5%) of the concentrate. Techno-functionality properties drastically differed according to the defatting solvent used and foaming properties were most affected. Thus, the techno-functional and whole properties must be considered for proper application of edible insects to achieve global food sustainability.

Naturally-derived protein extract from Gryllus bimaculatus improves antioxidant properties and promotes osteogenic differentiation of hBMSCs.

Naturally-derived proteins or peptides are promising biopolymers for tissue engineering applications owing to their health-promoting activity. Herein, we extracted proteins (~90%) from two-spotted cricket (Gryllus bimaculatus) and evaluated their osteoinductive potential in human bone marrow-derived mesenchymal stem cells (hBMSCs) under in vitro conditions. The extracted protein isolate was analyzed for the amino acid composition and the mass distribution of the constituent peptide fraction. Fourier transform infrared (FTIR) spectroscopy was used to determine the presence of biologically significant functional groups. The cricket protein isolate (CPI) exhibited characteristic protein peaks in the FTIR spectrum. Notably, an enhanced cell viability was observed in the presence of the extracted proteins, showing their biocompatibility. The CPI also exhibited antioxidant properties in a concentration-dependent manner. More significant mineralization was observed in the CPI-treated cells than in the control, suggesting their osteoinductive potential. The upregulation of the osteogenic marker genes (Runx2, ALP, OCN, and BSP) in CPI treated media compared with the control supports their osteoinductive nature. Therefore, cricket-derived protein isolates could be used as functional protein isolate for tissue engineering applications, especially for bone regeneration.

Protaetia brevitarsis seulensis Derived Protein Isolate with Enhanced Osteomodulatory and Antioxidative Property.

The osteogenic differentiation of stem cells is profoundly affected by their microenvironmental conditions. The differentiation behavior of stem cells can be tuned by changing the niche environments. The proteins or peptides that are derived by living organisms facilitate the osteogenic differentiation of stem cells. Here, we have evaluated the osteoinductive and antioxidative potential of the Protaetia brevitarsis seulensis insect-derived protein for human bone marrow-derived mesenchymal stem cells (hBMSCs). The amino acid contents in the isolated protein were determined by an amino acid analyzer. Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) were used to analyze the extract's functional groups and surface morphology. The extracted protein exhibited 51.08% ß-sheet conformation. No adverse effects were observed in extract-treated cells, indicating their biocompatibility. The protein isolate showed an excellent antioxidative property. Besides this, an enhancement in the hBMSCs' mineralization has been observed in the presence of treated protein isolates. Notably, osteogenic marker genes and proteins were effectively expressed in the treated cells. These results indicated that the P. brevitarsis-derived protein isolate can be used as a potential antioxidative biomaterial for bone tissue engineering applications.

Gravitational Effect on the Advancing and Receding Angles of a Two-Dimensional Cassie-Baxter Droplet on a Textured Surface.

Advancing and receding angles are physical quantities frequently measured to characterize the wetting properties of a rough surface. Thermodynamically, the advancing and receding angles are often interpreted as the maximum and minimum contact angles that can be formed by a droplet without losing its stability. Despite intensive research on wetting of rough surfaces, the gravitational effect on these angles has been overlooked because most studies have considered droplets smaller than the capillary length. In this study, however, by combining theoretical and numerical modeling, we show that the shape of a droplet smaller than the capillary length can be substantially modified by gravity under advancing and receding conditions. First, based on the Laplace pressure equation, we predict the shape of a two-dimensional Cassie-Baxter droplet on a textured surface with gravity at each pinning point. Then, the stability of the droplet is tested by examining the interference between the liquid surface and neighboring pillars and analyzing the free energy change upon depinning. Interestingly, it turns out that the apparent contact angles under advancing and receding conditions are not affected by gravity, while the overall shape of a droplet and the position of the pinning point are affected by gravity. In addition, the advancing and receding of the droplet with continuously increasing or decreasing volume are analyzed, and it is shown that the gravitational effect plays a key role in the movement of the droplet tip. Also, the gravitational effect on the degree of the stability of the droplet upon the external effect such as vibration is discussed. Finally, the theoretical predictions were validated against line tension-based front tracking modeling (LTM) that seamlessly captures the attachment and detachment between the liquid surface and the solid substrate. Our findings provide a deeper understanding on the advancing and receding phenomena of a droplet and essential insight into designing devices that utilize the wettability of rough surfaces.