Characterization of bacterial biofilms developed on the biodegradable polylactide and polycaprolactone polymers containing birch tar in an aquatic environment.

Birch tar was added to polylactide (PLA) and polycaprolactone (PCL) to create films with antimicrobial properties. After incubating the films for seven days in lake water, the diversity of bacterial communities developed on the surfaces of PCL and PLA with embedded birch tar (1 %, 5 %, and 10 %, w/w) was assessed with amplicon sequencing of the 16S rRNA gene on a MiSeq platform (Illumina). Notably, Aquabacterium and Caulobacter were more abundant at the surface of PCL compared to PLA (13.4 % vs 0.2 %, p < 0.001 and 9.5 % vs 0.2 %, p < 0.001, respectively) while Hydrogenophaga was significantly more abundant at the surface of PLA compared to PCL (6.1 % vs 1.8 %, p < 0.01). Overall, lower birch tar concentrations (1 % and 5 % on both polymers) stimulated bacterial diversity in biofilms compared to the control. The number of reeds assigned to Flavobacterium and Aquabacterium showed a rising trend with the increase of birch tar concentration on the surface of both polymers.

Application Potential of Trichoderma in the Degradation of Phenolic Acid-Modified Chitosan.

The aim of the study was to determine the potential use of fungi of the genus Trichoderma for the degradation of phenolic acid-modified chitosan in compost. At the same time, the enzymatic activity in the compost was checked after the application of a preparation containing a suspension of the fungi Trichoderma (spores concentration 105/mL). The Trichoderma strains were characterized by high lipase and aminopeptidase activity, chitinase, and ß-1,3-glucanases. T. atroviride TN1 and T. citrinoviride TN3 metabolized the modified chitosan films best. Biodegradation of modified chitosan films by native microorganisms in the compost was significantly less effective than after the application of a formulation composed of Trichoderma TN1 and TN3. Bioaugmentation with a Trichoderma preparation had a significant effect on the activity of all enzymes in the compost. The highest oxygen consumption in the presence of chitosan with tannic acid film was found after the application of the consortium of these strains (861 mg O2/kg after 21 days of incubation). Similarly, chitosan with gallic acid and chitosan with ferulic acid were found after the application of the consortium of these strains (849 mgO2/kg and 725 mg O2/kg after 21 days of incubation). The use of the Trichoderma consortium significantly increased the chitinase activity. The application of Trichoderma also offers many possibilities in sustainable agriculture. Trichoderma can not only degrade chitosan films, but also protect plants against fungal pathogens by synthesizing chitinases and ß-1,3 glucanases with antifungal properties.

Carboxymethyl cellulose based films enriched with polysaccharides from mulberry leaves (Morus alba L.) as new biodegradable packaging material.

The objective of this study was to determine the properties of a new active packaging film developed by the addition of mulberry leaves polysaccharides (MLP) into carboxymethyl cellulose (CMC). Biodegradable CMC-MLP films were fabricated by casting method with various concentrations of MLP (1, 5 and 10 % w/w). The addition of MLP into the CMC matrix resulted increased thickness (0.126 to 0.163 mm) and roughness of the films. Also, the decline in moisture content from 27.91 to 14.12 %, water vapor permeability from 8.95 to 5.21 × 10-10 g-1 s-1 Pa-1, and a swelling degree from 59.11 to 37.45 % were observed. With the increasing concentration of MLP, the mechanical properties of the films were improved and higher dispersion of UV light were noted. Fourier transform - infrared spectroscopy (FT-IR) and X-ray diffraction revealed good inter-molecular interaction between CMC matrix and MLP. The prepared films showed excellent thermal stability, antioxidant and antibacterial properties as well as susceptibility to biodegradation in the soil environment. Moreover, it was proved that the films have ability to retard oil oxidation. Overall, it was concluded that CMC-MLP films constitute a promising biomaterial that may be applied as active food packaging.

Technogenic soil salinisation, vegetation, and management shape microbial abundance, diversity, and activity.

The importance of the microbiome in the functioning of degraded lands in industrialised zones is significant. However, little is known about how environmental parameters affect microbial abundance, structure, diversity, and especially specific guilds involved in the nitrogen cycle in saline soils influenced by the soda industry. To address this knowledge gap, our research focused on assessing the microbiota in relation to soil properties and plant species composition across two transects representing different types of land use: saline wasteland and arable fields. Our findings show that the microbial communities were the most affected not only by soil salinity but also by pH and the composition of plant species. Taxonomic variability was the most shaped by salinity together with management type and CaCO3 content. The impact of salinity on the soil microbiome was manifested in a reduced abundance of bacteria and fungi, a lower number of observed phylotypes, reduced modularity, and a lower abundance of the nitrifying guild. Denitrification and nitrogen fixation were less affected by salinity. The last process was correlated with calcium carbonate. CaCO3 was also associated with microbial taxonomic variability and the overall microbial activity caused by hydrolases, which could aid organic matter turnover in saline but carbonate-rich sites. Bacterial genera such as Bacillus, Peanibacillus, and Rhodomicrobium, in addition to fungal taxa such as Cadophora, Mortierella globalpina, Preussia flanaganii, and Chrysosporium pseudomerdarium, show potential as favourable candidates for possible bioremediation initiatives. These results can be applied to future land reclamation projects. FUNDING INFORMATION: This research received no specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Bacillus paralicheniformis 2R5 and its impact on canola growth and N-cycle genes in the rhizosphere.

Chemical fertilization has a negative impact on the natural environment. Plant growth-promoting (PGP) rhizobacterial biofertilizers can be a safer alternative to synthetic agrochemicals. In this research, a culture-based method was used to assess the population size of rhizobacteria at the vegetative, flowering, and maturity stages of canola. Rhizobacteria were then isolated from each of the canola growth stages, and their seven PGP traits were determined. The highest abundance of culturable bacteria was found at the vegetative stage of the plants. Furthermore, four out of seven PGP traits were produced by the highest % of isolates at the vegetative stage. In the greenhouse experiment that included six rhizobacterial strains with best PGP traits, the greatest canola growth promotion ability under sterile conditions was observed after the introduction of Bacillus paralicheniformis 2R5. Moreover, under nonsterile conditions, 2R5 significantly increased canola growth. The presence of the trpA, B, C, D, E, F and pstA, and S genes in the 2R5 genome could be associated with canola growth promotion abilities. The chiA and mbtH genes could contribute to 2R5 antifungal activity against fungal pathogens. Moreover, the introduction of 2R5 significantly increased the abundance of the narG, nosZ, nifH, and nirS genes, which can prove that the 2R5 strain may be an important member of the soil bacterial community.

The effect of seed bacterization with Bacillus paralicheniformis 2R5 on bacterial and fungal communities in the canola rhizosphere.

Bacillus sp. is one of the best-studied plant growth-promoting rhizobacteria (PGPR). However, more detailed studies targeting its effect on the rhizosphere microbial community are required for improving management practices regarding its commercial application in the field. Our earlier study showed that PGPR Bacillus paralicheniformis 2R5 stimulated canola growth. Hence, this study aimed to assess the time-course impact of B. paralicheniformis 2R5 on bacterial and fungal community structure and diversity. The results showed that inoculation with B. paralicheniformis 2R5 initially significantly decreased the observed bacterial richness compared to the control, while after 44 days of treatment this alpha diversity metrics increased. A linear discriminant analysis effect size showed that B. paralicheniformis 2R5 altered the soil bacterial and fungal community structure by increasing the abundance of plants' beneficial microorganisms such as Nitrospira, Ramlibacter, Sphingomonas, Massilia, Terrimonas as well as Solicoccozyma, Schizothecium, Cyphellophora, Fusicolla, Humicola. B. paralicheniformis 2R5 seems to be a promising alternative to chemical pesticides and can be considered for practical application in the field. Its ability to alter the rhizosphere microbiome by increasing the diversity and composition of bacterial communities and increasing plants' beneficial groups of fungi, appears to be important in terms of improving canola development. However, further studies on these increased microbial taxa are necessary to confirm their function in promoting canola growth.

Pseudomonas sivasensis 2RO45 inoculation alters the taxonomic structure and functioning of the canola rhizosphere microbial community.

Inoculation with plant growth-promoting rhizobacteria (PGPR) is an eco-friendly sustainable strategy for improving crop productivity in diverse environments under different conditions. Our earlier study demonstrated that Pseudomonas sivasensis 2RO45 significantly stimulated canola (Brassica napus L. var. napus) growth. The aim of the present study was to investigate the structural and functional dynamics in the canola rhizosphere microbiome after inoculation with PGPR P. sivasensis 2RO45. The results based on alpha diversity metrics showed that P. sivasensis 2RO45 did not significantly alter the diversity of the native soil microbiota. However, the introduced strain modified the taxonomic structure of microbial communities, increasing the abundance of plant beneficial microorganisms, e.g., bacteria affiliated with families Comamonadaceae, Vicinamibacteraceae, genus Streptomyces, and fungi assigned to Nectriaceae, Didymellaceae, Exophiala, Cyphellophora vermispora, and Mortierella minutissima. The analysis of community level physiological profiling (CLPP) revealed that microbial communities in the P. sivasensis 2RO45 treated canola rhizospheres were more metabolically active than those in the non-treated canola rhizosphere. Four carbon sources (phenols, polymers, carboxylic acids, and amino acids) were better metabolized by the microbial communities from the rhizosphere of plants inoculated with the P. sivasensis 2RO45 than non-inoculated canola rhizospheres. Based on the community-level physiological profiles, the functional diversity of the rhizosphere microbiome was altered by the P. sivasensis 2RO45 inoculation. Substrate utilization Shannon diversity (H) index and evenness (E) index were significantly increased in the treated canola plants. The study provides new insight into PGPR-canola interactions for sustainable agriculture development.

Effect of birch tar embedded in polylactide on its biodegradation.

The plasticized film was made of polylactide and birch tar, which was used in a concentration of 1, 5 and 10 % by weight. Tar was added to the polymer to obtain materials with antimicrobial properties. The main purpose of this work is to characterize and biodegradation of this film after the end of its use. Therefore, the following analyzes were performed: enzymatic activity of microorganisms in the presence of polylactide (PLA) film containing birch tar (BT), biodegradation process in compost, barrier changes and structural properties of the film before and after biodegradation and bioaugmentation. Biological oxygen demand BOD21, water vapor permeability (Pv), oxygen permeability (Po), scanning electron microscopy (SEM) and enzymatic activity of microorganisms were assessed. Microorganism strains Bacillus toyonensis AK2 and Bacillus albus AK3 were isolated and identified, which constituted an effective consortium increasing the susceptibility of polylactide polymer material with tar to biodegradation in compost. Analyses with the use of the above-mentioned strains had an impact on the change of physicochemical properties, e.g. the presence of biofilm on the surface of the analyzed films and the reduction of the barrier properties of the film, which translates into the recorded susceptibility to biodegradation of these materials. The analyzed films can be used in the packaging industry, and after use, subjected to intentional biodegradation processes, including bioaugmentation.

Response of murine gut microbiota to a prebiotic based on oligosaccharides derived via hydrolysis of fungal α-(1→3)-d-glucan: Preclinical trial study on mice.

We investigated the modulating effect of α-(1→3)-glucooligosaccharides (GOS), i.e. a product of fungal α-(1→3)-d-glucan hydrolysis, on the gut microbiota composition. Mice were fed with a GOS-supplemented diet and two control diets for 21 days, and fecal samples were collected at 0, 1, and 3-week time points. The bacterial community composition was determined by 16S rRNA gene Illumina sequencing. The gut microbiota of the GOS-supplemented mice showed profound time-dependent changes in the taxonomic composition; however, we did not observe significant changes in α-diversity indices. The biggest number of genus abundance shifts after 1 week of the treatment was noticed between the group of the GOS-supplemented mice and the controls; however, the differences were still relevant after the 3-week treatment. The GOS-supplemented mice displayed higher abundance of Prevotella spp., with a concomitant decrease in the abundance of Escherichia-Shigella. Hence, GOS seems to be a promising candidate for a new prebiotic.

Physiological properties and genomic insights into the plant growth-promoting rhizobacterium Brevibacillus laterosporus K75 isolated from maize rhizosphere.

BACKGROUND: When looking for a safer alternative to pesticides that are potentially harmful to living organisms, one of the directions worth looking at are plant growth-promoting rhizobacteria. The purpose of the research was a comprehensive characterization of Brevibacillus laterosporus K75, a strain isolated from maize rhizosphere. Many studies have proved B. laterosporus to be a biocontrol agent; however, little is known about B. laterosporus as a plant growth-promoting rhizobacterium. RESULTS: Ninety strains were screened for plant growth-promoting activities. Four strains with the best plant growth-promoting traits (Rhodococcus qingshengii K8, Bacillus subtilis subsp. stercoris K73, Brevibacillus laterosporus K75, and Brevibacillus laterosporus K89) were used to research their effect on maize growth. Under sterile conditions, B. laterosporus K75 showed the best stimulatory effect, significantly improving the weight of roots, shoots and leaves, and considerably increasing content of chlorophyll. In unsterilized soil, B. laterosporus K75 significantly improved length of roots and weight of leaves compared to the K73, K89, and untreated control. Moreover, B. laterosporus K75 significantly increased specific leaf area compared to the untreated control and to other inoculant treatments. The genome of B. laterosporus K75 was compared to the recently published B. laterosporus MG64. Genome-mining displayed differences in identified plant growth-promoting genes and biosynthetic gene clusters of secondary metabolites. The B. laterosporus K75 genome possessed additional genes involved in indole-3-acetic acid production and phosphate solubilization that could be attributed to its ability to enhance maize growth. CONCLUSION: Our study demonstrated that B. laterosporus K75 is a promising candidate for use in inoculant formulation, effectively facilitating maize growth. © 2022 Society of Chemical Industry.

Plant growth-promoting rhizobacteria: Peribacillus frigoritolerans 2RO30 and Pseudomonas sivasensis 2RO45 for their effect on canola growth under controlled as well as natural conditions.

Even though canola is one of the most important industrial crops worldwide, it has high nutrient requirements and is susceptible to pests and diseases. Therefore, natural methods are sought to support the development of these plants. One of those methods could be a plant growth-promoting rhizobacteria (PGPR) that have a beneficial effect on plant development. The aim of this study was a genomic comparison of two PGPR strains chosen based on their effect on canola growth: Peribacillus frigoritolerans 2RO30, which stimulated canola growth only in sterile conditions, and Pseudomonas sivasensis 2RO45, which promoted canola growth in both sterile and non-sterile conditions. First of all, six bacterial strains: RO33 (Pseudomonas sp.), RO37 (Pseudomonas poae), RO45 (Pseudomonas kairouanensis), 2RO30 (Peribacillus frigoritolerans), 2RO45 (Pseudomonas sivasensis), and 3RO30 (Pseudomonas migulae), demonstrating best PGP traits in vitro, were studied for their stimulating effect on canola growth under sterile conditions. P. frigoritolerans 2RO30 and P. sivasensis 2RO45 showed the best promoting effect, significantly improving chlorophyll content index (CCI) and roots length compared to the non-inoculated control and to other inoculated seedlings. Under non-sterile conditions, only P. sivasensis 2RO45 promoted the canola growth, significantly increasing CCI compared to the untreated control and to other inoculants. Genome comparison revealed that the genome of P. sivasensis 2RO45 was enriched with additional genes responsible for ACC deaminase (acdA), IAA (trpF, trpG), and siderophores production (fbpA, mbtH, and acrB) compared to 2RO30. Moreover, P. sivasensis 2RO45 showed antifungal effect against all the tested phytopathogens and harbored six more biosynthetic gene clusters (BGC), namely, syringomycin, pyoverdin, viscosin, arylpolyene, lankacidin C, and enterobactin, than P. frigoritolerans 2RO30. These BGCs are well known as antifungal agents; therefore, it can be assumed that these BGCs were responsible for the antifungal activity of P. sivasensis 2RO45 against all plant pathogens. This study is the first report describing P. sivasensis 2RO45 as a canola growth promoter, both under controlled and natural conditions, thus suggesting its application in improving canola yield, by improving nutrient availability, enhancing stress tolerance, and reducing environmental impact of farming practices.

Polycaprolactone-Based Films Incorporated with Birch Tar-Thermal, Physicochemical, Antibacterial, and Biodegradable Properties.

We present new polymer materials consisting of polycaprolactone (PCL), polyethylene glycol (PEG), and birch tar (D). PEG was introduced into the polymer matrix in order to obtain a plasticizing effect, while tar was added to obtain antibacterial properties and to change the physicochemical properties of the film. The materials were obtained by the solvent method and characterized using a variety of methods to test their performance and susceptibility to biodegradation. The obtained data indicate that the introduction of the bioactive substance (D) into PCL improved the thermal stability and significantly lowered the Young's modulus values of the tested polymers. Moreover, the addition of birch tar improved the barrier and bacteriostatic properties, resulting in a reduction in the growth of pathogenic bacteria on the surface of the film. The films are not mutagenic but are susceptible to biodegradation in various environments. Due to their properties, they have potential for application in agriculture and horticulture and for packaging food, mainly vegetables grown in the field.

Consortium of plant growth-promoting rhizobacteria enhances oilseed rape (Brassica napus L.) growth under normal and saline conditions.

A preparation development, which stimulates plant growth under normal and saline conditions, and protects against fungal infections, would increase crop yields and reduce damage in agriculture. This study was conducted using bacterial isolates from rape rhizosphere as a plant growth promoter and an alternative to chemical fertilizers. Three from fifty bacterial isolates: B14 (Pseudomonas sp.), B16 (Sphingobacterium sp.), and B19 (Microbacterium sp.) showed the best in vitro plant growth-promoting (PGP) characteristics. B14 strain had the best antifungal activity against phytopathogens inhibiting growth of B. cinerea, C. acutatum, and P. lingam. Moreover, B14, B16 and B19 isolates coded for several genes involved in PGP activities, aimed at improving nutrient availability, resistance to abiotic stress, and fungal pathogen suppression. Microbial consortium (B14, B16, and B19) had the best effect on rape growth, significantly increasing number of live leaves, compared to the untreated control and single inoculant treatments. Moreover, the consortium induced significant increase in shoots length and chlorophyll content in comparison to Pseudomonas sp. B14 and Microbacterium sp. B19. The consortium also induced plants tolerance to salt stress. The genomic information as well as the observed traits, and beneficial attributes towards rape, make the rhizobacterial consortium an ideal candidate for further development as biofertilizers.

Comparative Nanopore Sequencing-Based Evaluation of the Midgut Microbiota of the Summer Chafer (Amphimallon solstitiale L.) Associated with Possible Resistance to Entomopathogenic Nematodes.

Root-feeding Amphimallon solstitiale larvae and certain other scarab beetles are the main soil-dwelling pests found in Europe, while entomopathogenic nematodes (EPN) have been used as a biocontrol agent against these species. Our study provides the first detailed characterization of the bacterial community of the midgut in wild A. solstitiale larvae, based on the nanopore sequencing of the 16S rRNA gene. In the whole dataset, we detected 2586 different genera and 11,641 species, with only 83 diverse bacterial genera shared by all studied individuals, which may represent members of the core midgut microbiota of A. solstitiale larvae. Subsequently, we compared the midgut microbiota of EPN-resistant and T0 (prior to EPN exposure) individuals, hypothesizing that resistance to this parasitic infection may be linked to the altered gut community. Compared to the control, the resistant insect microbiota demonstrated lower Shannon and Evenness indices and significant differences in the community structure. Our studies confirmed that the gut microbiota alternation is associated with resistant insects; however, there are many processes involved that can affect the bacterial community. Further research on the role of gut microbiota in insect-parasitic nematode interaction may ultimately lead to the improvement of biological control strategies in insect pest management.

Biodegradability of Novel Polylactide and Polycaprolactone Materials with Bacteriostatic Properties Due to Embedded Birch Tar in Different Environments.

The microbial biodegradation of new PLA and PCL materials containing birch tar (1-10% v/v) was investigated. Product of dry distillation of birch bark (Betula pendula Roth) was added to polymeric materials to obtain films with antimicrobial properties. The subject of the study was the course of enzymatic degradation of a biodegradable polymer with antibacterial properties. The results show that the type of the material, tar concentration, and the environment influenced the hydrolytic activity of potential biofilm degraders. In the presence of PCL films, the enzyme activities were higher (except for α-D-glucosidase) compared to PLA films. The highest concentration of birch tar (10% v/v) decreased the activity of hydrolases produced by microorganisms to the most significant extent; however, SEM analysis showed the presence of a biofilm even on plastics with the highest tar content. Based on the results of the biological oxygen demand (BOD), the new materials can be classified as biodegradable but, the biodegradation process was less efficient when compared to plastics without the addition of birch tar.

Microbial degradation of polyhydroxybutyrate with embedded polyhexamethylene guanidine derivatives.

The aim of this study was to isolate biofilm-forming bacteria that are capable of degrading polyhydroxybutyrate (PHB) with polyhexamethylene guanidine (PHMG) derivatives. The three types of derivatives incorporated in PHB and their concentration affected the biodegradability of the tested films in both water and compost. The PHMG derivative granular polyethylene wax at the highest concentration significantly inhibited BOD in both environments. At the same time, in water, PHB with PHMG stearate at 1% concentration was also found to inhibit biodegradation but to a lesser extent than PHMG polyethylene wax granulate. Analyzing the values of biofilm abundance and their hydrolytic activity in water, low concentrations of PHMG derivatives (0.2 and 0.6%) slightly inhibited biofilm abundance on the surface of the tested composites. Only granular polyethylene wax PHMG (at 1% concentration) significantly reduced biofilm formation and hydrolase activity in the compost to the greatest extent. Bacteria from biofilm were isolated and identified. Based on the 16S rRNA gene sequence, the strains belong to Bacillus toyonensis HW1 and Variovorax boronicumulans HK3. Introduction of the tested isolates to the environment can enhance composites degradation. However, this requires further research.

Nanopore-Sequencing Characterization of the Gut Microbiota of Melolontha melolontha Larvae: Contribution to Protection against Entomopathogenic Nematodes?

This study focused on the potential relationships between midgut microbiota of the common cockchafer Melolontha melolontha larvae and their resistance to entomopathogenic nematodes (EPN) infection. We investigated the bacterial community associated with control and unsusceptible EPN-exposed insects through nanopore sequencing of the 16S rRNA gene. Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes were the most abundant bacterial phyla within the complex and variable midgut microbiota of the wild M. melolontha larvae. The core microbiota was found to include 82 genera, which accounted for 3.4% of the total number of identified genera. The EPN-resistant larvae differed significantly from the control ones in the abundance of many genera belonging to the Actinomycetales, Rhizobiales, and Clostridiales orders. Additionally, the analysis of the microbiome networks revealed different sets of keystone midgut bacterial genera between these two groups of insects, indicating differences in the mutual interactions between bacteria. Finally, we detected Xenorhabdus and Photorhabdus as gut residents and various bacterial species exhibiting antagonistic activity against these entomopathogens. This study paves the way to further research aimed at unravelling the role of the host gut microbiota on the output of EPN infection, which may contribute to enhancement of the efficiency of nematodes used in eco-friendly pest management.

Exploring the properties of chitinolytic Bacillus isolates for the pathogens biological control.

Plant fungal diseases generate serious losses in the agriculture. The bacteria producing biologically active substances that inhibit the growth of fungal pathogens can be an alternative to the chemicals. The chitinolytic bacteria were isolated from the rhizosphere of wheat (Triticum aestivum L.) and their physiological properties which may be useful in the promotion of plant growth have been investigated. Their chitinases and antifungal activity were studied. The isolates were also tested for indirect growth-promoting traits such as ammonia production, siderophore production, hydrogen cyanide production, and salicylic acid production. Two chitinolytic strains B3 and B5 were identified as Bacillus subtilis and Bacillus sp., respectively. They produced active chitinases on a medium containing shrimp shell powder. The purified chitinases having the molecular weight of 35-45 kDa inhibited the growth of important plant pathogens such as Alternaria alternata, and Fusarium oxysporum. Additionally, the isolates showed the ability to produce a broad range of biological substances promoting the growth of plants.

Biofilm formation during biodegradation of polylactide, poly (3,4 hydroxybutyrate) and poly(ε-caprolactone) in activated sludge.

Biodegradable materials, namely pure polylactide (PLA), poly (3,4-hydroxybutyrate) (PHB), poly(ε-caprolactone) (PCL) were investigated to assess their degradability by activated sludge. The study aimed at the isolation of biofilm-forming bacteria and the determination of their hydrolytic activity toward the PLA, PHB, and PCL with embedded PHMG derivatives. The biological oxygen demand and physical properties (tensile strength, water vapor permeability, surface structure) of materials indicated that PCL was the best biodegradable film. Aeromonas and Rhodococcus isolated from the polymers' surface during the process of decomposition showed the ability to form biofilms. The introduction of PHMG derivatives into PLA, PCL, and PHB films did not affect biofilm formation and hydrolase activity for most of the isolates. PHMG derivatives at the concentration of 1% disturbed the degradation process.

Microbial Diversity in Deep-Subsurface Hot Brines of Northwest Poland: from Community Structure to Isolate Characteristics.

Deep-subsurface hot brines in northwest Poland, extracted through boreholes reaching 1.6 and 2.6 km below the ground surface, were microbiologically investigated using culture-independent and culture-dependent methods. The high-throughput sequencing of 16S rRNA gene amplicons showed a very low diversity of bacterial communities, which were dominated by phyla Proteobacteria and Firmicutes Bacterial genera potentially involved in sulfur oxidation and nitrate reduction (Halothiobacillus and Methylobacterium) prevailed in both waters over the sulfate reducers ("Candidatus Desulforudis" and Desulfotomaculum). Only one archaeal taxon, affiliated with the order Thermoplasmatales, was detected in analyzed samples. Bacterial isolates obtained from these deep hot brines were closely related to Bacillus paralicheniformis based on the 16S rRNA sequence similarity. However, genomic and physiological analyses made for one of the isolates, Bacillus paralicheniformis strain TS6, revealed the existence of more diverse metabolic pathways than those of its moderate-temperature counterpart. These specific traits may be associated with the ecological adaptations to the extreme habitat, which suggest that some lineages of B. paralicheniformis are halothermophilic.IMPORTANCE Deep-subsurface aquifers, buried thousands of meters down the Earth's crust, belong to the most underexplored microbial habitats. Although a few studies revealed the existence of microbial life at the depths, the knowledge about the microbial life in the deep hydrosphere is still scarce due to the limited access to such environments. Studying the subsurface microbiome provides unique information on microbial diversity, community structure, and geomicrobiological processes occurring under extreme conditions of the deep subsurface. Our study shows that low-diversity microbial assemblages in subsurface hot brines were dominated by the bacteria involved in biogeochemical cycles of sulfur and nitrogen. Based on genomic and physiological analyses, we found that the Bacillus paralicheniformis isolate obtained from the brine under study differed from the mesophilic species in the presence of specific adaptations to harsh environmental conditions. We indicate that some lineages of B. paralicheniformis are halothermophilic, which was not previously reported.