Search Results (19)

In this study, Ni²⁺ and Cd²⁺ resistant Pseudomonas aeruginosa 18, Enterobacter ludwigii 11Uz, and Enterobacter cloacae Uz_5 strains were isolated from soils contaminated with heavy metals in the Samarkand and Kashkadarya regions (Uzbekistan), and tested to remove Ni²⁺ and Cd²⁺ ions from the environment via biosorption. The biosorption capacity of these strains was observed under in vitro conditions. The biosorption process was highly dependent on the growing conditions, with the highest biosorption rate observed after 300 min of incubation at pH 7.0, and 40 °C. The presence of functional groups such as S=O, NH₂, and COOH in the biosorbing microorganisms was confirmed by IR spectroscopy. The adsorption capacity decreased when the initial metal concentration was increased and was enhanced with higher microbial biomass. Enterobacter ludwigii 11Uz strain was found to alter the toxic oxidation state of Ni²⁺ and Cd²⁺ cations, while Pseudomonas aeruginosa 18 and Enterobacter cloacae Uz_5 strains reduced the toxicity of Ni²⁺ cations only by changing their oxidation state. It was confirmed in our studies that the three selected bacterial strains actively participated in the detoxification of Cd²⁺ through the synthesis of cysteine amino acid. Full article

(1) Background: Xinjiang smoked horsemeat sausage is a popular food; however, bio-genic amine (BA) production is a concern for food safety. (2) Methods: the present study selected the three most toxic BAs for food safety (histamine, tyramine, and putrescine) and determined the bacteria that produce them. (3) Results: After 24 h of incubation, fifteen isolated strains, especially Enterobacter ludwigii MT705841 and Enterobacter bugandensis MT705832 produced putrescine (485.52 μg/mL and 408.95 μg/mL, respectively, p < 0.05); eight isolated strains, especially Proteus vulgaris MT705833 and Bacillus subtilis MT705839 produced histamine (63.86 μg/mL and 30.40 μg/mL, respectively, p < 0.05); and 14 isolated strains, especially Staphylococcus saprophyticus MT705831 and Proteus penneri MT705835 produced tyramine (482.26 μg/mL and 497.76 μg/mL, respectively, p > 0.05). Artemisia oil moderately inhibited P. vulgaris MT705833 and B. subtilis MT705839 after 48 h of in vitro incubation, decreasing histamine production by 44.83% and 47.92% for these two bacteria after 24 h and 20 h of incubation, respectively. Cinnamon oil strongly inhibited putrescine production by E. bugandensis MT705832 and E. ludwigii MT705841, decreasing production by 96.63% and 92.03% for these two bacteria after 24 h of incubation, respectively. Grapeseed oil slightly inhibited P. penneri MT705835 tyramine production (only after 4 h of incubation) and had an unstable inhibitory effect on Citrobacter freundii MT705836 tyramine production. (4) Conclusions: the results of this study suggest that cinnamon oil can be an effective food additive for the prevention of BA production in Xinjiang smoked sausages. Full article

The presence of opportunistic pathogens, such as the Enterobacter cloacae complex (ECC), in fresh vegetables poses a significant health risk, particularly amid the ongoing antibiotic resistance crisis. Traditional chemical decontamination methods are often ineffective and these are associated with issues such as cross-resistance between antibiotics and biocides, highlighting the need for alternative approaches. This study describes the isolation of a novel phage, FENT2, with anti-ECC activity, obtained from cattle farm sewage. Belonging to the Seunavirus genus, FENT2 did not carry genes associated with lysogenic cycle, antimicrobial resistance, or virulence factors. The phage demonstrated lytic activity against the host strain E. kobei AG07E, which harbored the mcr-9 gene, exhibiting a narrow host range that also included E. ludwigii strains. In vitro assays using BioTrac (SY-LAB) impedance technology confirmed the sustained lytic activity of FENT2 under food-related stress conditions, including pH levels from 5 to 7 and NaCl concentrations up to 2%. Furthermore, FENT2 demonstrated bactericidal potential on lettuce leaves, achieving 1 log reduction in bacterial counts of the host strain after 30 min immersion treatment. These findings highlight FENT2 as a promising candidate for biocontrol applications, offering a sustainable alternative to conventional decontamination methods for reducing antimicrobial-resistant ECC contamination in fresh produce. Full article

In this research study, we investigated four strains of Enterobacter ludwigii that showed promising properties for plant growth. These strains were tested for their ability to mobilize phosphorus and produce ammonium, siderophores, and phytohormones. The strains exhibited different values of PGP traits; however, the analysis of the complete genomes failed to reveal any significant differences in known genes associated with the expression of beneficial plant traits. One of the strains, GMG_278, demonstrated the best potential for promoting wheat growth in pot experiments. All morphological parameters of wheat were improved, both when GMG_278 was applied alone and when combined with mineral fertilizer. The combined effect we observed may suggest various mechanisms through which these treatments influence plants. The amount of pigments and proline suggests that bacterial introduction operates through pathways likely related to stress resilience. A study on the genetic mechanisms behind plant resilience to stress has revealed a significant upregulation of genes related to reactive oxygen species (ROS) defense after bacterial exposure. It is important to note that, in the initial experiments, the strain showed a significant production of salicylic acid, which is a potent inducer of oxidative stress. In addition, the synthesis of some phytohormones has been restructured, which may affect root growth and the architecture of root hairs. When combined with additional mineral fertilizers, these changes result in a significant increase in plant biomass. Full article

The rice leaf folder Cnaphalocrocis medinalis is an important migratory pest in Asia. Although this pest possesses diverse bacterial communities in its gut, functions of these bacteria in modulating host fitness, including development durations, pupal weight, adult longevity, and fecundity, remain unknown. We isolated gut bacteria from field–collected C. medinalis larvae using a culture–dependent method and identified 15 bacterial isolates. Six of the isolates (Klebsiella aerogenes, Klebsiella pneumoniae, Enterobacter ludwigii, Enterobacter asburiae, Pantoea dispersa, and Pantoea ananatis) were newly discovered in C. medinalis. When larvae were orally inoculated with individual bacterial isolates, 15 isolates showed varying degrees of effects on C. medinalis fitness. Importantly, we found that 10 bacterial isolates induced significant larval mortality. Specifically, the inoculation of Pseudomonas mosselii, P. dispersa, Chryseobacterium culicis, P. ananatis, and Myroides odoratus caused high mortality ranging from 40.0% to 56.7%. However, reducing the entire gut bacterial community with antibiotic treatment negatively impacted C. medinalis fitness, while the reinoculation of a bacterial community to antibiotic–treated larvae recovered some of the adverse effects. In particular, control and bacterial community–inoculated C. medinalis laid approximately 37.6% more eggs than antibiotic–treated C. medinalis. This suggests that these bacteria affect their hosts differently when they are together as compared to alone. Our results reveal that C. medinalis harbors gut bacteria capable of both mutualistic and pathogenic interactions, suggesting their potential as biocontrol agents and indicating that targeting the gut bacterial community could be an effective strategy for controlling C. medinalis infestations. Full article

This study explores the efficacy of bio-efficient solutions, specifically plant growth-promoting microorganisms (PGPMs), in sustainable soil management. This research was conducted in 2020. It evaluates the impact of various single microbial inoculants, including Enterobacter ludwigii, Bacillus subtilis, Pseudomonas fluorescens, Kosakonia cowanii, and Trichoderma harzianum, on plant growth soil enzyme activity and organism abundance. Perennial ryegrass and mustard were used as test plants, in controlled environmental conditions. The results show generally positive effects of microbial inoculants on plant biomass (E. ludwigii increased ryegrass biomass by 9.75%, and P. fluorescens increased mustard biomass by up to 38.81% compared to the control) and on soil microbial activities. Our study further investigated the combined application of all these strains in five different soil types and textures. The results highlight the significance of soil physicochemical properties in determining inoculant efficacy; we found that clayey soils with higher colloid content support more robust microbial activity. Additionally, using natural clay minerals like alginite for enhancing soil conditions showed promising interactions with microbial inoculants, although application requires further optimization. These findings suggest that integrating microbial inoculants in sustainable agricultural practices could enhance plant growth, improve soil health, and reduce the need of chemical fertilizers. Future research should aim to refine the combinations and application methods of these bio-efficient solutions for broader agricultural applicability. Full article

We analyzed the whole genome sequences (WGS) and antibiograms of 35 Enterobacter isolates, including E. hormaechei and E. asburiae, and the recently described E. bugandensis, E. kobei, E. ludwigii, and E. roggenkampii species. Isolates were obtained from human blood and urinary tract infections in patients in the United States. Our goal was to understand the genetic diversity of antimicrobial resistance genes and virulence factors among the various species. Thirty-four of 35 isolates contained an AmpC class bla_ACT allele; however, the E. roggenkampii isolate contained bla_MIR-5. Of the six Enterobacter isolates resistant to ertapenem, imipenem, and meropenem, four harbored a carbapenemase gene, including bla_KPC or bla_NDM. All four isolates were mCIM-positive. The remaining two isolates had alterations in ompC genes that may have contributed to the resistance phenotype. Interpretations of cefepime test results were variable when disk diffusion and automated broth microdilution results were compared due to the Clinical Laboratory and Standards Institute use of the “susceptible dose-dependent” classification. The diversity of the bla_ACT alleles paralleled species identifications, as did the presence of various virulence genes. The classification of recently described Enterobacter species is consistent with their resistance gene and virulence gene profiles. Full article

Plant biostimulants have received attention as sustainable alternatives to chemical fertilizers. Extracellular polymeric substances (EPSs), among the compounds secreted by plant growth-promoting rhizobacteria (PGPRs), are assumed to alleviate abiotic stress. This study aims to investigate the effect of purified EPSs on rice under abiotic stress and analyze their mechanisms. A pot experiment was conducted to elucidate the effects of inoculating EPSs purified from PGPRs that increase biofilm production in the presence of sugar on rice growth in heat-stress conditions. Since all EPSs showed improvement in SPAD after the stress, Enterobacter ludwigii, which was not characterized as showing higher PGP bioactivities such as phytohormone production, nitrogen fixation, and phosphorus solubilization, was selected for further analysis. RNA extracted from the embryos of germinating seeds at 24 h post-treatment with EPSs or water was used for transcriptome analysis. The RNA-seq analysis revealed 215 differentially expressed genes (DEGs) identified in rice seeds, including 139 up-regulated and 76 down-regulated genes. A gene ontology (GO) enrichment analysis showed that the enriched GO terms are mainly associated with the ROS scavenging processes, detoxification pathways, and response to oxidative stress. For example, the expression of the gene encoding OsAAO5, which is known to function in detoxifying oxidative stress, was two times increased by EPS treatment. Moreover, EPS application improved SPAD and dry weights of shoot and root by 90%, 14%, and 27%, respectively, under drought stress and increased SPAD by 59% under salt stress. It indicates that bacterial EPSs improved plant growth under abiotic stresses. Based on our results, we consider that EPSs purified from Enterobacter ludwigii can be used to develop biostimulants for rice. Full article

The present study examined the biosynthesis and characterization of selenium nanoparticles (SeNPs) using two contrasting endophytic selenobacteria, one Gram-positive (Bacillus sp. E5 identified as Bacillus paranthracis) and one Gram-negative (Enterobacter sp. EC5.2 identified as Enterobacter ludwigi), for further use as biofortifying agents and/or for other biotechnological purposes. We demonstrated that, upon regulating culture conditions and selenite exposure time, both strains were suitable “cell factories” for producing SeNPs (B-SeNPs from B. paranthracis and E-SeNPs from E. ludwigii) with different properties. Briefly, dynamic light scattering (DLS), transmission electron microscopy (TEM), and atomic force microscopy (AFM) studies revealed that intracellular E-SeNPs (56.23 ± 4.85 nm) were smaller in diameter than B-SeNPs (83.44 ± 2.90 nm) and that both formulations were located in the surrounding medium or bound to the cell wall. AFM images indicated the absence of relevant variations in bacterial volume and shape and revealed the existence of layers of peptidoglycan surrounding the bacterial cell wall under the conditions of biosynthesis, particularly in the case of B. paranthracis. Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), energy-dispersive X-ray (EDS), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS) showed that SeNPs were surrounded by the proteins, lipids, and polysaccharides of bacterial cells and that the numbers of the functional groups present in B-SeNPs were higher than in E-SeNPs. Thus, considering that these findings support the suitability of these two endophytic stains as potential biocatalysts to produce high-quality Se-based nanoparticles, our future efforts must be focused on the evaluation of their bioactivity, as well as on the determination of how the different features of each SeNP modulate their biological action and their stability. Full article

A study on the ability of new microbial strains to assimilate biodiesel-derived glycerol at low purity (75% w/w) and produce extra-cellular platform chemical compounds of major interest was carried out. After screening several bacterial strains under different fermentation conditions (e.g., pH, O₂ availability, glycerol purity), three of the screened strains stood out for their high potential to produce valued-added products such as 2,3-butanediol (BDO), 1,3-propanediol (PDO) and ethanol (EtOH). The results indicate that under aerobic conditions, Klebsiella oxytoca ACA-DC 1581 produced BDO in high yield (Y_BDO/Gly = 0.46 g/g, corresponding to 94% of the maximum theoretical yield; Y_mt) and titer, while under anaerobic conditions, Citrobacter freundii NRRL-B 2645 and Enterobacter ludwigii FMCC-204 produced PDO (Y_PDO/Gly = 0.56 g/g, 93% of Y_mt) and EtOH (Y_EtOH/Gly = 0.44 g/g, 88% of Y_mt), respectively. In the case of C. freundii, the regulation of pH proved to be mandatory, due to lactic acid production and a subsequent drop of pH that resulted in fermentation ceasing. In the fed-batch culture of K. oxytoca, the BDO maximum titer reached almost 70 g/L, the Y_BDO/Gly and the mean productivity value (Pr_BDO) were 0.47 g/g and 0.4 g/L/h, respectively, while no optimization was imposed. The final BDO production obtained by this wild strain (K. oxytoca) is among the highest in the international literature, although the bioprocess requires optimization in terms of productivity and total cost. In addition, for the first time in the literature, a strain from the species Hafnia alvei (viz., Hafnia alvei ACA-DC 1196) was reported as a potential BDO producer. The strains as well as the methodology proposed in this study can contribute to the development of a biorefinery that complements the manufacture of biofuels with high-value biobased chemicals. Full article

The restoration of submerged macrophytes is commonly limited by abiotic stress. Here, we isolated PGPR strains from the rhizosphere of submerged macrophytes using ACC deaminase selective medium, and evaluated their growth promoting effects on Vallisneria natans (V. natans) under low light intensity and (or) high sediment organic matter load, and also explored the indigenous microbiome response of V. natans seedlings to PGPR inoculants. Ten isolates were chosen from the 252 isolated strains based on the ACC deaminase activity and the production of IAA and siderophore. M1 (Pseudomonas vancouverensis) and E15 (Enterobacter ludwigii) had the best growth promoting effects under low light stress and under double stress of low light and high sediment organic matter load, and the shoot height increased by 36% and 46%, respectively. The results of indigenous microbiome analysis showed that PGPR inoculants could regulate the relative abundance of unclassified_f_Enterobacteriaceae and improve the α-diversity of the rhizosphere bacterial community. Under high sediment organic matter load, inoculation of PGPR obviously shifted the β-diversity of rhizosphere bacterial communities to promote the early growth of V. natans. This study expands the application of plant–microbe interaction in the field of freshwater ecological restoration. Full article

This study analyzed the resistome, virulome and mobilome of an MCR-9-producing Enterobacter sp. identified in a muscle sample of seabream (Sparus aurata), collected in a land tank from multitrophic fish farming production. Average Nucleotide Identity analysis identified INSAq77 at the species level as an Enterobacter ludwigii INSAq77 strain that was resistant to chloramphenicol, florfenicol and fosfomycin and was susceptible to all other antibiotics tested. In silico antimicrobial resistance analyses revealed genes conferring in silico resistance to β-lactams (bla_ACT-88), chloramphenicol (catA4-type), fosfomycin (fosA2-type) and colistin (mcr-9.1), as well as several efflux pumps (e.g., oqxAB-type and mar operon). Further bioinformatics analysis revealed five plasmid replicon types, including the IncHI2/HI2A, which are linked to the worldwide dissemination of the mcr-9 gene in different antibiotic resistance reservoirs. The conserved nickel/copper operon rcnR-rcnA-pcoE-ISSgsp1-pcoS-IS903-mcr-9-wbuC was present, which may play a key role in copper tolerance under anaerobic growth and nickel homeostasis. These results highlight that antibiotic resistance in aquaculture are spreading through food, the environment and humans, which places this research in a One Health context. In fact, colistin is used as a last resort for the treatment of serious infections in clinical settings, thus mcr genes may represent a serious threat to human health. Full article

Fish gut represents a peculiar ecological niche where bacteria can transit and reside to play vital roles by producing bio-compounds with nutritional, immunomodulatory and other functions. This complex microbial ecosystem reflects several factors (environment, feeding regimen, fish species, etc.). The objective of the present study was the identification of intestinal microbial strains able to produce molecules called biosurfactants (BSs), which were tested for surface and antibacterial activity in order to select a group of probiotic bacteria for aquaculture use. Forty-two bacterial isolates from the digestive tracts of twenty Mediterranean grey mullets were screened for testing emulsifying (E-24), surface and antibiotic activities. Fifty percent of bacteria, ascribed to Pseudomonas aeruginosa, Pseudomonas sp., P. putida and P. anguilliseptica, P. stutzeri, P. protegens and Enterobacter ludwigii were found to be surfactant producers. Of the tested strains, 26.6% exhibited an antibacterial activity against Staphylococcus aureus (10.0 ± 0.0–14.5 ± 0.7 mm inhibition zone), and among them, 23.3% of isolates also showed inhibitory activity vs. Proteus mirabilis (10.0 ± 0.0–18.5 ± 0.7 mm inhibition zone) and 6.6% vs. Klebsiella pneumoniae (11.5 ± 0.7–17.5 ± 0.7 mm inhibition zone). According to preliminary chemical analysis, the bioactive compounds are suggested to be ascribed to the class of glycolipids. This works indicated that fish gut is a source of bioactive compounds which deserves to be explored. Full article

The food industry is still searching for novel solutions to effectively ensure the microbiological safety of food, especially fresh and minimally processed food products. Nowadays, the use of bacteriophages as potential biological control agents in microbiological food safety and preservation is a promising strategy. The aim of the study was the isolation and comprehensive characterization of novel bacteriophages with lytic activity against saprophytic bacterial microflora of minimally processed plant-based food products, such as mixed leaf salads. From 43 phages isolated from municipal sewage, four phages, namely Enterobacter phage KKP 3263, Citrobacter phage KKP 3664, Enterobacter phage KKP 3262, and Serratia phage KKP 3264 have lytic activity against Enterobacter ludwigii KKP 3083, Citrobacter freundii KKP 3655, Enterobacter cloacae KKP 3082, and Serratia fonticola KKP 3084 bacterial strains, respectively. Transmission electron microscopy (TEM) and whole-genome sequencing (WGS) identified Enterobacter phage KKP 3263 as an Autographiviridae, and Citrobacter phage KKP 3664, Enterobacter phage KKP 3262, and Serratia phage KKP 3264 as members of the Myoviridae family. Genome sequencing revealed that these phages have linear double-stranded DNA (dsDNA) with sizes of 39,418 bp (KKP 3263), 61,608 bp (KKP 3664), 84,075 bp (KKP 3262), and 148,182 bp (KKP 3264). No antibiotic resistance genes, virulence factors, integrase, recombinase, or repressors, which are the main markers of lysogenic viruses, were annotated in phage genomes. Serratia phage KKP 3264 showed the greatest growth inhibition of Serratia fonticola KKP 3084 strain. The use of MOI 1.0 caused an almost 5-fold decrease in the value of the specific growth rate coefficient. The phages retained their lytic activity in a wide range of temperatures (from −20 °C to 50 °C) and active acidity values (pH from 4 to 11). All phages retained at least 70% of lytic activity at 60 °C. At 80 °C, no lytic activity against tested bacterial strains was observed. Serratia phage KKP 3264 was the most resistant to chemical factors, by maintaining high lytic activity across a broader range of pH from 3 to 11. The results indicated that these phages could be a potential biological control agent against saprophytic bacterial microflora of minimally processed plant-based food products. Full article

Drought stress is a prevalent environmental stress that adversely affects agricultural industries worldwide. In this study, bacterial isolates, AFFR02 and Mj1212, showed tolerance to polyethylene glycol-induced (PEG) drought stress (approximately 15%) and possess strong phosphate-solubilizing capacity. Moreover, we investigated the plant growth attributes, chlorophyll content, and ion uptake in alfalfa plants (Medicago sativa L) inoculated with isolates AFFR02 and Mj1212 under drought stress. We observed that drought stress drastically affects alfalfa’s growth attributes: shoot length: SL (24.88%), root length: RL (29.62%), shoot fresh weight: SFW (49.62%), root fresh weight: RFW (45.09%), stalk diameter: SD (52.84%), and chlorophyll content: CC (19.2%). However, in bacterial-inoculated alfalfa plants, the growth attributes significantly recovered were SL (12.42%), RL (21.30%), SFW (50.74%), RFW (46.42%), SD (76.72%), and CC (17.98%). In drought-stressed alfalfa plants, we observed a significant decrease in the relative water content (7.45%), whereas there was an increase in electrical conductivity (68.87%) and abscisic acid contents (164.42%). Antioxidant analysis showed a significant increase in total phenolic content (46.08%), DPPH-scavenging activity (39.66%), total flavonoid (13.68%), and superoxide dismutase (28.51%) in alfalfa treated with drought stress and bacterial isolates AFFR02 and Mj1212 simultaneously. Moreover, an increase in inductively coupled plasma (ICP) analysis of potassium (17.98%), phosphorous (11.14%), calcium (3.07%), and magnesium (6.71%) was recorded for bacteria-inoculated alfalfa plants under drought stress. In conclusion, bacterial isolates AFFR02 and Mj1212 enhance alfalfa growth under drought stress. Therefore, the isolates could be used as potential candidates in smart-climate agricultural practices in drought-stricken areas worldwide. Full article