Search Results (110)

This study aimed to isolate, characterize, and evaluate bacterial consortia capable of degrading the diamide insecticide cyantraniliprole in aqueous systems and to assess their bioremediation potential under environmentally relevant conditions. Four bacterial consortia, each comprising six isolates, demonstrated significant growth in mineral media containing cyantraniliprole as the sole carbon source, and the isolates were identified using conventional microbiological techniques in combination with MALDI-TOF-MS analysis. The bacterial consortia were enriched from pesticide-contaminated environments and systematically evaluated using microbiological, physiological, and analytical approaches to determine their degradation potential and environmental adaptability. The degradation performance of the consortia was systematically assessed under varying environmental parameters, including temperature, pH, salinity, and incubation time, with optimal degradation observed at 30–35 °C, pH 7.0–8.0, 0.5–5.0% NaCl, and 11 days of incubation at 150 rpm using an initial cyantraniliprole concentration of 50 mg/L. Biodegradation efficiency was further evaluated using DCPIP reduction assays, alongside measurements of biofilm formation and biomass production, indicating enhanced metabolic activity and adaptive responses under pesticide-induced stress. The consortia also exhibited the capacity to degrade structurally related diamide pesticides, including flubendiamide, chlorantraniliprole, cyclaniliprole, and fluchlordiniliprole, suggesting broad-spectrum biodegradation potential. Their performance was further validated in a simulated water microcosm system designed to mimic environmentally relevant contamination scenarios. In simulated contaminated water (60 mg/L cyantraniliprole), bacterial inoculants standardized to 10⁷ CFU/mL achieved substantial degradation after 20 days of incubation at 30 °C, as confirmed by HPLC analysis, with the six-strain consortium (T4), comprising Bacillus subtilis subsp. subtilis AZFS3, Bacillus pumilus AZFS5, Bacillus mojavensis AZFS15, Bacillus paramycoides AZFS18, Pseudomonas aeruginosa KZFS4, and Alcaligenes aquatilis KZFS11, demonstrating the highest removal efficiency (98.27%) and reducing the pesticide concentration to 1.00 mg/L, followed by consortium T3 (96.72%), which consisted of Bacillus subtilis Ht1, Bacillus subtilis Ht2, Bacillus mojavensis Ht3, Pseudomonas aeruginosa Ht4, Pseudomonas aeruginosa Ht5, and Pseudomonas aeruginosa Ht6. Residue analysis and predictive bioinformatic assessment further supported the biodegradation capacity of the selected bacterial communities and suggested the formation of simpler transformation products. Overall, the investigated bacterial consortia exhibited high degradation efficiency and environmental adaptability, highlighting their potential as effective and eco-friendly agents for the bioremediation of cyantraniliprole-contaminated water systems. Full article

Pneumonia remains a leading cause of morbidity and mortality worldwide, with bacterial pathogens contributing significantly to its burden. While Pseudomonas aeruginosa and Acinetobacter baumannii complex are well-recognized non-fermenting Gram-negative bacteria (NFGNB) causing severe pneumonia, particularly in healthcare settings, an expanding array of other, rarer NFGNB species is increasingly implicated. These species include, but are not limited to, Achromobacter spp., Ochrobactrum spp., Burkholderia spp., Aeromonas spp., Roseomonas spp., Elizabethkingia spp., Chryseobacterium spp. Alcaligenes spp., Ralstonia spp., Cupriavidus spp., Sphingomonas spp., Rhizobium spp., Empedobacter spp., and Brevundimonas spp. In this article we aim to provide a focused review of the contemporary epidemiology and specific risk factors for pneumonia caused by this diverse group of rare NFGNB, explicitly excluding P. aeruginosa, Stenotrophomonas maltophilia, and A. baumannii. We seek to delineate the emerging patterns of pneumonia associated with Achromobacter spp., Burkholderia spp., Aeromonas spp., Roseomonas spp., Elizabethkingia spp., Pandoraea spp., Sphingomonas spp., and K. gyiorum. Moreover, we discuss antimicrobial treatment strategies for pneumonia caused by rarer NFGNB including Ochrobactrum spp., Chryseobacterium spp., Alcaligenes spp., Ralstonia spp., Cupriavidus spp., Rhizobium spp., Empedobacter spp., and Brevundimonas spp. A deeper understanding of these specific epidemiological trends and risk factors is important for guiding precise diagnostic approaches, informing antimicrobial stewardship programs, and developing targeted infection prevention and control strategies with the aim of mitigating the impact of these challenging pathogens in the clinical setting. Full article

The novel strain TD-94 with higher ammonia degradation efficiency was isolated from the activated sludge of SINOPEC and belongs to the family of Alcaligenes faecalis (A. faecalis) based on its 16sRNA sequence and physio-biochemical characteristics. It is a Gram-negative, highly heterotrophic aerobic ammoxidation bacterium that is capable of effectively treating ammonia-nitrogen wastewater. The genome size of strain TD-94 was 4,361,949 bp with a GC content of 56.47% and a total of 4101 genes, which accounted for 89.54% of the total genome length. Analysis of various databases showed that 649 genes were annotated in the GO database; a total of 2712, 4095 and 12 genes were annotated in the KEGG, COG, and ADRB databases, respectively; and there were 24 types of cytochrome P450, 477 signal peptides, and eight secondary metabolites. All these data provide a theoretical basis for the mechanism of action of the strain TD-94. Based on the whole-genome sequencing results, functional genes related to nitrogen metabolism in A. faecalis TD-94, including aerobic ammonia oxidation (AOB), hydroxylamine oxidoreductase (HAO), and pyruvic oxime dioxygenase (POD) were identified. Through growth curve analysis and the identification of functional genes, the nitrogen metabolism pathway of A. faecalis TD-94 was proposed, demonstrating that the strain TD-94 has good denitrification capabilities and a novel degradative pathway. Full article

Nicotinoproteins are a group of NAD⁺-dependent dehydrogenases that bind NAD⁺ tightly and catalyze reactions without using free NAD⁺. In this study, we investigated the role of the unique insertion loop in nicotinoproteins. Carveol dehydrogenase (CADh), a short-chain dehydrogenase/reductase (SDR) nicotinoprotein, and β-hydroxybutyrate dehydrogenase from Alcaligenes faecalis (AfBHBDh), a non-nicotinoprotein counterpart, were used as model enzymes. An insertion loop-deleted mutant, CADh Δ39–49, was constructed. An insertion loop from Mycobacterium paratuberculosis CADh (MpCADh) was introduced into AfBHBDh to generate the two mutants. The results showed that CADh Δ39–49 lost NAD⁺ tight binding capacity and could not utilize free NAD⁺. In contrast, the AfBHBDh mutants showed no dye-mediated dehydrogenase activity. Moreover, the K_M and K_D values for NAD⁺ were higher than those of the wild-type enzyme. Docking simulations revealed a stronger binding affinity between NAD⁺ and the mutants than with the wild-type AfBHBDh. Taken together, these results suggest that the insertion loop interferes with NAD⁺ entry into the active site of the enzyme while creating a more energetically favorable binding environment. This loop is necessary but alone is insufficient to achieve NAD⁺ tight binding. This study deepens understanding of NAD⁺ binding in SDR nicotinoproteins and provides insights for SDR enzyme engineering. Full article

This study addresses the pressing issue of high-ammonia nitrogen wastewater, such as landfill leachate, by developing immobilized microbial beads that combine high mechanical strength with efficient denitrification performance. The beads were prepared using a composite of sodium alginate (SA), carboxymethyl cellulose (CMC), and diatomite (DE), with a dual-ion (Ca²⁺-Al³⁺) stepwise cross-linking technique to encapsulate Alcaligenes faecalis. The material ratios were systematically optimized through single-factor and response surface methodology (RSM), identifying the optimal conditions as: SA 2.0%, CMC 1.5%, DE 1.0%, CaCl₂ 2.25%, and Al₂(SO₄)₃ 2.0%. Under these conditions, the beads achieved a mechanical strength of 3.20 N and exhibited an ammonia nitrogen removal rate of 93.10% after 96 h of treating actual landfill leachate (NH₃-N ≈ 1000 mg/L). In conclusion, the SA-CMC-DE dual-ion cross-linked beads demonstrate structural stability and efficient mass transfer, offering an economically viable and novel solution for the treatment of high-ammonia nitrogen wastewater. Full article

Cadmium (Cd) contamination in agricultural soils severely impairs plant growth, disrupts microbial communities, and threatens food safety due to its high toxicity and mobility. Conventional remediation methods are often expensive and environmentally unsustainable. In contrast, plant–microbiome interactions offer an eco-friendly solution to reduce Cd accumulation and improve plant growth. Arbuscular mycorrhizal fungi (AMF) and mycorrhiza helper bacteria (MHB) are known to improve plant growth and resilience in Cd-contaminated soils. However, the mechanisms by which AMF and MHB co-inoculation could reduce soil Cd contamination by altering the rhizosphere fungal community remain unclear. This study aimed to evaluate how co-inoculation with AMF (Funneliformis mosseae) and MHB (Alcaligenes faecalis) affects plant Cd uptake and soil Cd content, and how it reshapes the cucumber rhizosphere fungal community. A greenhouse experiment was conducted with four treatments: CK (no inoculation), Fm (AMF inoculation), Af (MHB inoculation), and FA (AMF + MHB co-inoculation). Co-inoculation with AMF and MHB (FA) significantly reduced Cd concentrations in both plant tissues and soil. Fungal communities were profiled using Illumina MiSeq sequencing of the ITS region, and diversity metrics and structural changes were assessed through PCoA and DESeq2. Co-inoculation (FA) significantly reshaped the fungal community, increasing the relative abundances of beneficial phyla such as Mortierellomycota, Basidiomycota and Glomeromycota, while decreasing the abundance of potentially pathogenic Ascomycota. Double inoculation with AMF and MHB also enhanced fungal diversity, as measured by the Simpson index, and enriched specific OTUs. This study uncovers the mechanisms through which AMF–MHB co-inoculation reduces Cd concentrations in both plants and soil by altering the cucumber rhizosphere fungal community composition. These findings demonstrate that AMF–MHB co-inoculation is an effective, biologically driven strategy for remediating Cd-contaminated soils by restructuring cucumber rhizosphere fungal communities. Full article

Lipid A is a component of lipopolysaccharide (LPS) of Gram-negative bacteria. Previously, we demonstrated that synthesized lipid A derived from Alcaligenes faecalis (ALA) could enhance antigen-specific immunoglobulin (Ig) A and T helper (Th) 17 responses, when ALA was co-administered experimentally with an antigen as a vaccine adjuvant. This raised concerns about the safety of the ALA usage, since IgA and Th17 responses have been suggested to play a pathogenic role in several immune-mediated diseases, including multiple sclerosis (MS). We investigated whether ALA administrations could exacerbate an animal model of MS, Theiler’s murine encephalomyelitis virus (TMEV) infection. TMEV-infected SJL/J mice were administered ALA at various time points, and their neurological signs were observed for 7 weeks. We found that ALA administrations did not exacerbate TMEV-induced inflammatory disease or viral persistence in the central nervous system (CNS), clinically or histologically. Furthermore, ALA administrations did not enhance TMEV-specific humoral and cellular responses, including IgA and Th17 responses. On the other hand, principal component analysis (PCA) of the fecal, not the ileal, samples showed significant changes in the microbiota, characterized by increases in the relative abundance of bacteria belonging to the phylum Bacteroidota, including the genera Alistipes and Bacteroides. Therefore, ALA injections could be safe for use in immune-mediated diseases, whose immunopathology has been associated with IgA and Th17 responses. Full article

Background/Objectives: Antimicrobial resistance (AMR) in wildlife is an emerging public health concern due to the risk of zoonotic transmission, especially through the food chain, yet data on free-ranging animals remain scarce. This study examined the presence and patterns of AMR among bacteria isolated from hunted wild boars in the Campania region of Italy. Methods: Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) was used to identify bacterial isolates from wild boar meat and carcass swabs to the species level, and the Kirby–Bauer disk diffusion test was applied to screen 205 isolates, spanning 20 bacterial genera, against a panel of clinically relevant antibiotics. Resistance metrics were analyzed at genus and antibiotic levels, and patterns were visualized using a hierarchically clustered heatmap. Results: Resistance was detected in 15 of the 20 genera, with full susceptibility observed in Acinetobacter, Arthrobacter, Glutamicibacter, Leclercia, and Rahnella. Overall, 67.3% (138/205) of the isolates showed resistance to at least one antibiotic, with 33.7% (69/205) classified as multidrug-resistant (MDR). Carbapenems retained the highest activity (≥95% susceptibility) among all genera tested, while amoxicillin/clavulanate (78.4%) and aztreonam (57.4%) exhibited the highest mean resistance. Among potential pathogens, Escherichia coli exhibited an extended-spectrum β-lactamase (ESBL)-like phenotype, with resistance to amoxicillin/clavulanate (67%), aztreonam (54%), and ceftazidime (47%) but preserved carbapenem susceptibility. Staphylococcus spp. showed pronounced resistance to linezolid (57%) and erythromycin (52%), whereas Pseudomonas isolates demonstrated elevated resistance to aztreonam and ceftazidime (57% each). Opportunistic pathogens such as Alcaligenes faecalis and Pantoea agglomerans showed peak resistance to ciprofloxacin and amoxicillin/clavulanate. Pathogens and opportunistic pathogens demonstrated higher mean resistance (>30%) than commensals (≤32%), but the difference in mean and median resistance levels was not statistically significant (Mann–Whitney’s U test, W = 4, p = 0.39). Conclusions: These findings highlight the widespread occurrence of AMR and MDR phenotypes, with clinically significant resistance patterns in wild-boar-associated bacteria, including non-pathogenic strains, highlighting their role in the amplification of AMR. Although the preservation of carbapenem susceptibility underscores their potential as last-line antibiotics, the high resistance to commonly used antibiotics raises concerns for zoonotic transmission. Surveillance of wildlife reservoirs therefore remains critical for integrated AMR control. Full article

Nitrogen pollution in wastewater remains a pressing environmental concern, prompting the need for efficient and sustainable treatment technologies. This study constructs an HN-AD microbial consortium using three pre-screened strains (Delftia tsuruhatensis SDU2, Pseudomonas stutzeri SDU10, Alcaligenes faecalis SDU20) and evaluates its bioaugmentation effects in sequencing batch reactors (SBRs). An orthogonal test optimized the inoculation ratio as 2:3:3 (SDU2:SDU10:SDU20), achieving the highest ammonium removal efficiency of 96.02% in vitro. In SBR experiments, the bioaugmented reactor (SBR1) demonstrated superior nitrogen and organic matter removal compared to the control (SBR2). By day 40 of the enhancement phase, SBR1 achieved 88.9% ammonium removal and 93.7% COD removal, representing improvements of 20.5% and 17.9% over SBR2, respectively. Microbial community analysis revealed that bioaugmentation enriched Proteobacteria and Bacteroidota phyla, promoting functional guilds like Alcaligenes, Pseudomonas, and Paracoccus, which synergized with indigenous microbiota to enhance metabolic efficiency. This study highlights the potential of HN-AD consortia to overcome limitations of conventional nitrogen removal systems, offering a promising strategy for optimizing wastewater treatment processes. Full article

Background: Chronic and acute wounds are often colonized by polymicrobial biofilms, delaying healing and complicating treatment. Rapid, non-invasive detection of pathogenic bacteria is therefore crucial for timely and targeted therapy. This study investigated porphyrin-producing bacterial species using the handheld cureVision imaging system. Methods: In this study, 20 clinically relevant, porphyrin-producing bacterial species were cultured on δ-aminolevulinic acid (ALA)-supplemented agar and analyzed using the handheld cureVision imaging system under 405 nm excitation. Both Red-Green-Blue (RGB) and fluorescence images were acquired under ambient daylight conditions, and fluorescence signals were quantified by grayscale intensity analysis. Results: All tested species exhibited measurable red porphyrin-associated fluorescence, with the highest intensities observed in Klebsiella pneumoniae, Klebsiella oxytoca, Veillonella parvula, and Alcaligenes faecalis. A standardized detectability threshold of 0.25, derived from negative controls, enabled semi-quantitative comparison across species. Statistical analysis confirmed that the fluorescence intensities of all bacterial samples were significantly elevated compared to the control (Wilcoxon signed-rank test and sign test, both p < 0.001; median intensity = 0.835, IQR: 0.63–0.975). Conclusions: These results demonstrate that the cureVision system enables robust and reliable detection of porphyrin-producing wound bacteria, supporting its potential as a rapid, non-invasive diagnostic method for assessing wound colonization and guiding targeted clinical interventions. Full article

Increasing demand for high-quality food is driving the development of biologized farming methods, which involve the use of microorganisms, including endophytes, to stimulate plant growth. However, research on the composition of endosphere microbiomes is limited. The study presents an analysis of the bacterial endophytic microbiome in lettuce seeds (Lactuca sativa L., cv. Ozornik) using high-throughput sequencing of 16S rRNA amplicons. It evaluates the taxonomic composition and putative functional properties of seed endophytic bacteria. The microbial community exhibited low diversity (Shannon index ranged from 1.1 to 1.84, Simpson index from 0.57 to 0.83). The bacterial endophytic community of lettuce seeds was dominated by Pseudomonadota (83%), Actinomycetota (14%), and Bacillota (3%). The genera identified within the microbiome included Pantoea (32%), Rhodococcus (13%), Candidatus Profftella (13%), Janthinobacterium (7%), Pseudomonas (9%), Enterococcus (3%), and Alcaligenes (2%), which exhibit a broad spectrum of beneficial properties: plant growth promotion (PGPB), suppression of phytopathogens, enhanced stress tolerance, participation in contaminant biodegradation, and heavy metal detoxification. The structure and functional potential of the microbiome vary between samples, potentially due to differences in source material and cultivation conditions. The obtained results expand our understanding of the composition and functions of endophytic bacteria in lettuce seeds, which is important for the development of novel biocontrol agents for plants consumed by humans in an unprocessed form. Full article

Poly(ε-caprolactone) (PCL) is a synthetic plastic known for its excellent physicochemical properties and a wide range of applications in packaging, coatings, foaming, and agriculture. In medicine, its versatility allows it to function as a scaffold for drug delivery, sutures, implants, tissue engineering, and 3D printing. In addition to its biocompatibility, PCL’s most notable characteristic is its biodegradability. However, this property is affected by temperature, microbial activity, and environmental conditions, which means PCL can sometimes remain in nature for long periods. This review shows that various types of microorganisms can efficiently degrade PCL, including different strains of Pseudomonas spp., Streptomyces spp., Alcaligenes faecalis, and fungi like Aspergillus oryzae, Fusarium spp., Rhizopus delemar, and Thermomyces lanuginosus. These microorganisms produce enzymes such as lipases, esterases, and cutinases that break down PCL into smaller molecules that act as substrates. The review also examines the phylogenetic diversity of organisms capable of biodegrading PCL, the biochemical pathways involved in this process, and specific aspects of the genetic framework responsible for the expression of the enzymes that facilitate degradation. Targeted research on microbial PCL biodegradation and its practical applications could significantly aid in reducing and managing plastic waste on a global ecological scale. Full article

4-Nonylphenols (4-NPs) are persistent endocrine disruptors frequently found in wastewater treatment plant (WWTP) effluents and sewage sludge. This study evaluated the ability of eight bacterial strains that were isolated from sewage sludge to degrade 4-n-NP in an aqueous solution. Bacillus safensis CN12, Shewanella putrefaciens CN17, and Alcaligenes faecalis CN8 showed the highest degradation rates, removing 100%, 75%, and 74% of 4-n-NP (10 mg L⁻¹), with DT₅₀ values of 0.90, 8.9, and 10.4 days, respectively. Despite the reduction in 4-n-NP concentrations, ecotoxicity assays revealed that the resulting transformation products (TPs) were more toxic than the parent compound. To investigate the potential degradation mechanisms, in silico and gene expression analyses were conducted on B. safensis CN12, revealing a significant upregulation of the multicopper oxidase gene, cotA (7.25-fold), and the ring-cleaving dioxygenase gene, mhqO (13.9-fold). Although the CN12 strain showed potential for mineralization based on gene expression studies, this was not observed in the aqueous solution. However, when 4-n-NP was adsorbed on sludge and treated with CN12 in the presence of hydroxypropyl-β-cyclodextrin (HPBCD) as a bioavailability enhancer, mineralization reached up to 33%, indicating a synergistic effect with the native sludge microbiota. Full article

The genus Terminalia has a long history of use in traditional medicine to treat various diseases, including bacterial infections. We previously reported a metabolomic analysis using liquid chromatography–mass spectrometry of selected Australian Terminalia spp. and highlighted numerous flavonoids that may contribute to the antimicrobial activities of those plants. This study examines the antibacterial activities of fifteen flavonoids found in Terminalia spp. against a range of gastrointestinal pathogens using broth dilution assays. Flavonoids were also combined with six different classes of conventional antibiotics to investigate interactions. The efflux pump inhibitory activity of the flavonoid was evaluated using ethidium bromide accumulation and efflux assays. Toxicities were assessed via human dermal fibroblast cell line assays. Fisetin, hispidulin, isoorientin, orientin, rutin, and vitexin showed noteworthy growth inhibitory activity (MIC values 62.5–250 µg/mL). Isoorientin and orientin were most potent against Bacillus cereus and Alcaligenes faecalis, displaying MIC values of 62.5 µg/mL against both bacteria. All flavonoids except genistein, isorhamnetin, kaempferol, luteolin, taxifolin, and vitexin were nontoxic in human dermal fibroblast (HDF) cell proliferation assays. When individual flavonoids were combined with selected antibiotics, some potentiated the activity of these antibiotics. Two synergistic, eighteen additive and thirty-one non-interactive interactions were observed. The synergistic interactions were all observed in combination with orientin. Notably, orientin exhibited efflux pump inhibitory effects at concentrations from 15.26 µg/mL to 125 µg/mL. The findings reported herein indicate that the selected flavonoids have the potential for addressing bacterial antibiotic resistance and highlight the need for further study. Full article

Processing of meat waste into meat and bone meal generates wastewater that must be properly treated to minimize its environmental impact. In addition to its high organic load, it contains microorganisms, including pathogens, that pose a threat to human health. In this study, wastewater from meat and bone meal production was treated using the Fenton process with a variable Fe²⁺/H₂O₂ mass ratio ranging from 1:2 to 1:10, followed by neutralization with lime milk. A microbiological analysis was performed on both the raw wastewater and the liquid fractions obtained after treatment, along with additional analyses of selected physicochemical parameters. The identification of isolated microorganisms was carried out using the MALDI-TOF MS technique. Qualitative analysis of the treated wastewater samples showed the presence of microorganisms belonging to the genera Staphylococcus, Enterococcus, Alcaligenes, and Pseudomonas. Staphylococcus aureus and Alcaligenes faecalis were present in each of the treated samples. The Fenton process effectively reduced the total number of microorganisms, with the lowest counts of 1.3 × 10² CFU/mL and 3.2 × 10² CFU/mL found in wastewater samples treated with Fe²⁺/H₂O₂ mass ratios of 1:8 and 1:10, which correlated with higher doses of hydrogen peroxide. Full article