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Sugar beet is the most important crop for sugar production in temperate zones. The plant microbiome is considered an important factor in crop productivity and health. Here, we investigated the bacterial diversity of seeds, roots, and rhizosphere of five sugar beet hybrids named Eduarda (ED), Koala (KO), Tibor (T), Tajfun (TF), and Cercospora-resistant (C). A culture-independent next-generation sequencing approach was used for the further investigation of seed-borne endophytes. Hybrid-associated bacteria were evaluated for their plant growth–promoting (PGP) characteristics, antagonistic activity towards Cercospora beticola and several Fusarium strains in dual culture assays, and drought and salinity tolerance. High-throughput sequencing revealed that the Proteobacteria phylum was most dominant in the seeds of all hybrids, followed by Cyanobacteria and Actinobacteriota. The predominant genus in all hybrids was Pantoea, followed by Pseudomonas, Acinetobacter, Chalicogloea, Corynebacterium, Enterobacter, Enterococcus, Glutamicibacter, Kosakonia, and Marinilactibacillus. Unique genera in the hybrids were Pleurocapsa and Arthrobacter (T), Klebsiella (TF), Apibacter (ED), and Alloscardovia (KO). The genera that were most represented in one hybrid were Weissella and Staphylococcus (TF); Streptococcus (T); Gardnerella, Prevotella, and Rothia (KO); and Gilliamella, Lactobacillus, and Snodgrassella (ED). Thirty-two bacteria out of 156 isolates from the rhizosphere, roots, and seeds were selected with respect to various plant growth–promoting activities in vitro, i.e., nitrogen fixation, phosphate solubilization, siderophore production, indole-3-acetic acid production, 1-aminocyclopropane-1-carboxylic acid deaminase activity, hydrogen cyanide production, exoenzymatic activity (amylase, protease, lipase, cellulase, xylanase, mannanases, gelatinase, and pectinase), mitigation of environmental stresses, and antifungal activity. Mixta theicola KO3-44, Providencia vermicola ED3-10, Curtobacterium pusillum ED2-6, and Bacillus subtilis KO3-18 had the highest potential to promote plant growth due to their multiple abilities (nitrogen fixation, phosphate solubilization, production of siderophores, and IAA). The best antagonistic activity towards phytopathogenic fungi was found for Bacillus velezensis C3-19, Paenibacillus polymyxa C3-36 and Bacillus halotolerans C3-16/2.1. Only four isolates B. velezensis T2-23, B. subtilis T3-4, B. velezensis ED2-2, and Bacillus halotolerans C3-16/2.1 all showed enzymatic activity, with the exception of xylanase production. B. halotolerans C3-16/2.1 exhibited the greatest tolerance to salinity, while two B. subtilis strains (C3-62 and TF2-1) grew successfully at the maximum concentration of PEG. The current study demonstrates that sugar beet–associated bacteria have a wide range of beneficial traits and are therefore highly promising for the formulation of biological control and PGP agents. [ABSTRACT FROM AUTHOR]

Chironomus riparius are sediment-dwelling invertebrates in freshwater ecosystems and are used as indicators of environmental pollution. Their habitat is threatened by high levels of contaminants such as microplastics and organic matter. A promising strategy for the eco-friendly degradation of pollutants is the use of bacteria and their enzymatic activity. The aim of this study was to characterize for the first time bacteriobiota associated with the gut of C. riparius larvae from nature and laboratory samples, to compare it with sediment and food as potential sources of gut microbiota, and to assess its ability to degrade cellulose, proteins, and three different types of microplastics (polyethylene, polyvinyl chloride, and polyamide). The metabarcoding approach highlighted Proteobacteria, Firmicutes, Bacteroidota, and Actinobacteriota as most abundant in both gut samples. Culturable microbiota analysis revealed Metabacillus idriensis, Peribacillus simplex, Neobacillus cucumis, Bacillus thuringiensis/toyonensis, and Fictibacillus phosphorivorans as five common species for nature and laboratory samples. Two P. simplex and one P. frigoritolerans isolates showed the ability for intensive growth on polyethylene, polyvinyl chloride, and polyamide. Both cellulolytic and proteolytic activity was observed for Paenibacillus xylanexedens and P. amylolyticus isolates. The characterized strains are promising candidates for the development of environmentally friendly strategies to degrade organic pollution and microplastics in freshwater ecosystems. [ABSTRACT FROM AUTHOR]

The primary purpose of the study, as part of the planned conservation work, was to uncover all aspects of autochthonous biofilm pertaining to the formation of numerous deterioration symptoms occurring on the limestone Rožanec Mithraeum monument in Slovenia. Using state-of-the-art sequencing technologies combining mycobiome data with observations made via numerous light and spectroscopic (FTIR and Raman) microscopy analyses pointed out to epilithic lichen Gyalecta jenensis and its photobiont, carotenoid-rich Trentepohlia aurea, as the origin of salmon-hued pigmented alterations of limestone surface. Furthermore, the development of the main deterioration symptom on the monument, i.e., biopitting, was instigated by the formation of typical endolithic thalli and ascomata of representative Verrucariaceae family (Verrucaria sp.) in conjunction with the oxalic acid-mediated dissolution of limestone. The domination of lichenized fungi, as the main deterioration agents, both on the relief and surrounding limestone, was additionally supported by the high relative abundance of lichenized and symbiotroph groups in FUNGuild analysis. Obtained results not only upgraded knowledge of this frequently occurring but often overlooked group of extremophilic stone heritage deteriogens but also provided a necessary groundwork for the development of efficient biocontrol formulation applicable in situ for the preservation of similarly affected limestone monuments. [ABSTRACT FROM AUTHOR]

The olive tree is one of the most important agricultural plants, affected by several pests and diseases that cause a severe decline in health status leading to crop losses. Olive leaf spot disease caused by the fungus Venturia oleaginea can result in complete tree defoliation and consequently lower yield. The aim of the study was to obtain new knowledge related to plant–pathogen interaction, reveal mechanisms of plant defense against the pathogen, and characterize fungal phyllosphere communities on infected and symptomless leaves that could contribute to the development of new plant breeding strategies and identification of novel biocontrol agents. The highly susceptible olive variety "Istrska Belica"' was selected for a detailed evaluation. Microscopy analyses led to the observation of raphides in the mesophyll and parenchyma cells of infected leaves and gave new insight into the complex V. oleaginea pathogenesis. Culturable and total phyllosphere mycobiota, obtained via metabarcoding approach, highlighted Didymella, Aureobasidium, Cladosporium, and Alternaria species as overlapping between infected and symptomless leaves. Only Venturia and Erythrobasidium in infected and Cladosporium in symptomless samples with higher abundance showed statistically significant differences. Based on the ecological role of identified taxa, it can be suggested that Cladosporium species might have potential antagonistic effects on V. oleaginea. [ABSTRACT FROM AUTHOR]

Biocontrol strains from the genera Bacillus and Pseudomonas and their bioactive secondary metabolites may be considered beneficial for plant health, since they act through several mechanisms, including direct antibiosis, plant growth promotion and the induction of systemic resistance in the plant hosts. These beneficial bacteria have been proven to be very active against bacterial and fungal phytopathogens, nematodes and different insects. Direct antibiosis, an induced immune system response (ISR) in the host plant and competition for nutrients and space are the most common biocontrol potential of these genera. Species belonging to the Bacillus genus are commonly considered ideal due to their rapid growth, ease of handling and excellent colonizing properties. These endospore‐forming Bacillus strains are capable of suppressing and inhibiting plant pathogens, both indirectly by competing with the pathogens for a niche or nutrient requirements, or directly by producing various lipopeptide compounds such as iturin, surfactin and fengycin, which are active on many plant pathogens. Furthermore, they are also capable of inducing systemic resistance in plants through the production of volatile substances including alcohols, aldehydes, aromatics, sulfides and ketones. Lipopeptides, polyketides and volatiles from Bacillus spp. can stimulate the expression of genes coding for pathogenesis-related (PR) proteins and other defense-related proteins in the plant hosts through the activation of jasmonic acid (JA), salicylic acid (SA) or ethylene (ET) signaling pathways. In addition, inhibition of quorum sensing in competitive bacteria and the capability to downregulate expression of genes involved in mycelial growth, penetration, sporulation and the virulence of a fungal pathogen is another property of Bacillus strains and their volatiles. The Pseudomonas genus is rich in species with the potential for biocontrol with positive effects on plant welfare, which actively participate in complex plant-pathogen-antagonist interaction. The most common molecules involved in this mechanism are, among others, 2,4‐diacetylphloroglucinol, phenazine‐1‐carboxylic acid, phenazine-1-carboxamide, pyoluteorin and pyrrolnitrin. Cyclic lipopeptides from Pseudomonas spp. such as nunamycin, nunapeptin, brasmycin and braspeptin were intensively studied as agents for plant biocontrol and biostimulation in agriculture. Nunamycin, nunapeptin, brasmycin and braspeptin were identified as essential in the antifungal role. Furthermore, phenazines, sessilins and orfamides were shown to have additive roles in the suppression of some fungal diseases. Additionally, Bacillus and Pseudomonas spp. produce chitinases, glucanases and proteases involved in the suppression of many fungal diseases. Their production is mainly induced by the presence of fungal pathogen biomass and their cell wall. This review provides an updated overview of the antimicrobial activity of plant-associated Bacillus and Pseudomonas involved in plant disease suppression via biological control mechanisms, including their molecular basis and direct activity, offering a better understanding in preventing different pests. • Bacillus and Pseudomonas spp. produce wide array of secondary metabolites. • Secondary metabolites are involved in biocontrol and biofertilization mechanisms. • Numerous phytopathogens are controlled by Bacillus and Pseudomonas isolates. • Molecular mechanisms of biocontrol activity are discussed. [ABSTRACT FROM AUTHOR]

Bacterial metabarcoding analysis by 16S rDNA of five occurrences of mineral waters in Serbia (Torda, Slankamen Banja, Lomnički Kiseljak, Velika Vrbnica and Obrenovačka Banja) indicated the presence of a high percentage of the Proteobacteria phylum, followed by the Bacteroidetes phylum. The families Rhodobacteraceae, Burkholderiaceae, Pseudomonadaceae, Methylophilaceae and Moraxellaceae were the most dominant in the bacterial flora of the selected occurrences, whereas the most represented genera were Acinetobacter, Pseudorhodobacter, Pseudomonas, Limnohabitans, Massilia, Limnobacter and Methylotenera. The presence of coliform bacteria was not detected. Alpha diversity analysis revealed that Slankamen Banja and Lomnički Kiseljak were the richest of the selected occurrences, while the mineral waters of Torda, Velika Vrbnica and Obrenovačka Banja were characterized by similar diversity of bacterial communities determined by beta diversity analysis. Physical-chemical analysis revealed the value of total dissolved solids above 1 g/L, as well as elevated concentrations of some metals and non-metals. The research concluded that specific bacteria contribute to the development of biocorrosion and biofouling processes of water intake facilities. In addition, some of these bacteria might be potential indicators of the organic sources of pollution and/or biotechnological natural remediators in the treatment of contaminated waters. [ABSTRACT FROM AUTHOR]

Bats as flying mammals are potent vectors and natural reservoir hosts for many infectious viruses, bacteria, and fungi, also detected in their excreta such as guano. Accelerated deforestation, urbanization, and anthropization hastily lead to overpopulation of the bats in urban areas allowing easy interaction with other animals, expansion, and emergence of new zoonotic disease outbreaks potentially harmful to humans. Therefore, getting new insights in the microbiome of bat guano from different places represents an imperative for the future. Furthermore, the use of novel high-throughput sequencing technologies allows better insight in guano microbiome and potentially indicated that some species could be typical guano-dwelling members. Bats are well known as a natural reservoir of many zoonotic viruses such as Ebola, Nipah, Marburg, lyssaviruses, rabies, henipaviruses, and many coronaviruses which caused a high number of outbreaks including ongoing COVID-19 pandemic. Additionally, many bacterial and fungal pathogens were identified as common guano residents. Thus, the presence of multi-drug-resistant bacteria as environmental reservoirs of extended spectrum β-lactamases and carbapenemase-producing strains has been confirmed. Bat guano is the most suitable substrate for fungal reproduction and dissemination, including pathogenic yeasts and keratinophilic and dimorphic human pathogenic fungi known as notorious causative agents of severe endemic mycoses like histoplasmosis and fatal cryptococcosis, especially deadly in immunocompromised individuals. This review provides an overview of bat guano microbiota diversity and the significance of autochthonous and pathogenic taxa for humans and the environment, highlighting better understanding in preventing emerging diseases. Key points: Bat guano as reservoir and source for spreading of autochthonous and pathogenic microbiota Bat guano vs. novel zoonotic disease outbreaks Destruction of bat natural habitats urgently demands increased human awareness [ABSTRACT FROM AUTHOR]

Extensive employment of plant resins and propolis, as an antiseptic agents dating from ancient times in numerous cultures indicating that it may have antimicrobial and other biological properties. Resins from deciduous trees from the Populus and Salix genera, several fruit trees from the genus Prunus and a few other species were analyzed for their phenolic composition and antibacterial activity. Phenolic profiling of the plant resins was performed by high-performance thin-layer chromatography (HPTLC) and ultra-high-performance liquid chromatography (UHPLC) coupled with hybrid mass spectrometry. Antimicrobial activity against seven bacterial species was determined by minimum inhibitory concentration (MIC) assays and bioautography. The synergism, additivism, and antagonism of phenolic compounds were used to define the nature and type of interactions. Populus spp. showed higher amounts of p -hydroxybenzoic acid, p -coumaric acid, caffeic acid, chrysin, apigenin, quercetin, pinocembrin, pinobanksin and galangin, which confirmed the botanical origin of the orange and blue types of propolis. In addition, Gram-positive bacteria exhibited high susceptibility to poplar samples while being resistant to samples from other origins. Cherry bud samples had high amounts of naringenin and showed strong activity against Bacillus subtilis and Listeria monocytogenes . The combinations of tested phenolics showed mainly additive or indifferent effects. [ABSTRACT FROM AUTHOR]

The composition of essential oil (EO) from different aerial parts of Artemisia absinthium L. (Asteraceae), from Serbia, was analyzed. Shoots without leaves and inflorescences (SWLI), leaves (L), and inflorescences (I) were subjected to hydrodistillation using the Clevenger-type apparatus. Analysis of EO was done by gas chromatography with MS of flame ionization detection (GC/MS and GC/FID). Microdilution method was used to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Organoleptic characteristics and quantitative and qualitative differences were documented between examined EOs. In total, 17 compounds were identified in SWLI, 11 in L and 18 in I. The principal constituent in all EOs was trans-thujone (SWLI - 23.56%, L - 60.41%, and I - 46.16%). The most susceptible strains were Xanthomonas campestris pv. campestris, Erwinia amylovora and Rathayibacter tritici. The strongest activity was shown for SWLI EO against E. amylovora (0.03 mg/mL). The same MIC values were observed for L EO against E. amylovora and R. tritici (0.09 mg/ mL). The equal activity for all EOs tested was detected against X. campestris (0.13 mg/mL). [ABSTRACT FROM AUTHOR]

Due to the increasing number of bacterial infections and the development of resistivity toward antibiotics, new materials and approaches for treatments must be urgently developed. The production of new materials should be ecologically friendly considering overall pollution with chemicals and economically acceptable and accessible to the wide population. Thus, the possibility of using biocompatible graphene quantum dots (GQDs) as an agent in photodynamic therapy was studied. First, dots were obtained using electrochemical cutting of graphite. In only one synthetic step using gamma irradiation, GQDs were doped with N atoms without any reagent. Obtained dots showed blue photoluminescence, with a diameter of 19–89 nm and optical band gap of 3.23–4.73 eV, featuring oxygen-containing, amino, and amide functional groups. Dots showed antioxidative activity; they quenched •OH at a concentration of 10 μg·mL−1, scavenged DPPH• radicals even at 5 μg·mL−1, and caused discoloration of KMnO4 at 30 μg·mL−1. Under light irradiation, dots were able to produce singlet oxygen, which remained stable for 10 min. Photoinduced effects by GQDs were studied on several bacterial strains (Listeria monocytogenes, Bacillus cereus, clinical strains of Streptococcus mutans, S. pyogenes, and S. sangunis, Pseudomonas aeruginosa, and one yeast strain Candida albicans) but antibacterial effects were not noticed. [ABSTRACT FROM AUTHOR]

• Thirteen N,N -disubstituted chloroacetamides were synthesized as covalent inhibitors • Several chloroacetamides effectively activated both extrinsic and intrinsic apoptotic pathways • 3D QSAR and pharmacophoric similarity search suggested alcohol-metabolizing enzymes as promising targets • Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) identified as potential anticancer targets through pharmacophoric similarity search. • Docking studies demonstrated hydrogen bonds and hydrophobic interactions with ADH and ALDH. The recent FDA approval of afatanib, ibrutinib, and osimertinib, which covalently bind to specific cysteine residues in target kinases, has renewed interest in covalent drug discovery. Besides α,β -unsaturated carbonyls, chloroacetamides have emerged as popular warheads for designing targeted covalent inhibitors. In this study, we synthesized thirteen N,N -disubstituted chloroacetamides (1 – 13) by acylating secondary amines with chloroacetyl chloride, selecting substituents to provide a wide range of lipophilicity. We evaluated their anticancer and antimicrobial activity, finding five compounds with significant cytotoxicity against HeLa, K562, and A549 cell lines (IC 50 <10 μM). Notably, compound 10 activated caspases 3, 8, and 9, promoting both intrinsic and extrinsic apoptotic pathways, while compounds 9 – 12 were strong apoptosis inducers. A 3D QSAR model showed that aromatic substituents on nitrogen atoms reduced HeLa potency, whereas the overall molecular shape had a positive effect. ChemBL and pharmacophoric similarity searches suggested potential anticancer targets, including alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), glycogen synthase kinase-3β, and calmodulin. Docking studies indicated that chloroacetamides bind to ADH and ALDH via hydrogen bonds and hydrophobic interactions. Pharmacokinetics predictions suggest that chloroacetamides are druglike molecules with promising ADMET properties. [ABSTRACT FROM AUTHOR]

The composition of essential oil from aerial parts of Ambrosia artemisiifolia L. from Bor (Serbia) was analyzed. The essential oil was obtained by hydrodistillation and analyzed by gas chromatography (GC-FID, GC-MS). In total, 45 compounds were detected (98.49% of the total). The essential oil was dominated by monoterpene (45%) and sesquiterpene (38.51%) hydrocarbons. The principal constituents were germacrene D (25.3%), limonene (21.6%), and α-pinene (15.7%). The microdilution method was used to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the essential oil against five Gram-negative phytopathogenic strains. Essential oil exhibited strong antimicrobial activity against two Xanthomonas campestris strains and one referent and one natural isolate of Ervinia amylovora, causative agents of black rot and fire blight. [ABSTRACT FROM AUTHOR]

In this work, we demonstrate that the rhizosphere of common duckweed (Lemna minor) is inhabited with various phenol-resistant bacterial strains. Based on 16S rRNA sequencing, we have identified 60 rhizosphere-associated bacterial isolates belonging to 10 different bacterial genera (Pseudomonas, Hafnia, Serratia, Enterobacter, Micrococcus, Stenotrophomonas, Xanthomonas, Bacillus, Staphylococcus and Klebsiella). All isolates have been tested for phenol resistance and ability to utilize phenol as the sole carbon source. 70% of all isolates survived high doses of phenol (≥200 mg/L) and at least 27% can be potentially acclimatized by gradual increase of phenol concentration. Finally, based on high phenol resistance, ability to utilize phenol as the sole carbon source and documented low pathogenicity, we propose 5 strains as potentially excellent candidates for bioremediation. These 5 strains taxonomically correspond to Klebsiella sp., Serratia sp., and Hafnia sp., respectively. To the best of our knowledge, this is the first attempt to assess decontamination capacity of Serratia nematodiphila and Hafnia sp. in the context of bioremediation of phenol-contaminated aqueous media. Although additional analyses are needed, interaction between the common duckweed and the selected bacterial strains may be utilized in future bioremediation strategies. [ABSTRACT FROM AUTHOR]

Identification of 33 Bacillus spp. isolates from different environmental samples collected from the territory of Serbia was performed by sequencing of the 5'-hypervariable section of 16S rRNA gene. Eight species were identified within four phylogenetic groups: B. pumilus, B. megaterium, B. subtilis and B. cereus. Determination of their antibiotic resistance was performed using the minimum inhibitory concentration (MIC) assay. We found that just one isolate was resistant to gentamicin, 9 were resistant to clindamycin and all were resistant to vancomycin. Based on the profile of resistance, the isolates were categorized into 4 categories. In silico analysis of the erythromycin-resistance (erm) gene for clindamycin resistance showed their distribution between related and nonrelated soil and human isolates including different species of Bacillus genera. This finding indicates that Bacillus spp. from the environment could be a source of resistance to clindamycin. The potential for the presence and spread of resistance determinants in the soil and similar ecosystems exists so that monitoring of antibiotic resistance genes in nonpathogenic Bacillus strains from the environment is advised. [ABSTRACT FROM AUTHOR]