Enterobacter Strains Research Articles

Effect of endophytic diazotroph Enterobacter roggenkampii ED5 on nitrogen-metabolism-related microecology in the sugarcane rhizosphere at different nitrogen levels.

Sugarcane is an important sugar and energy crop worldwide, requiring a large amount of nitrogen (N). However, excessive application of synthetic N fertilizer causes environmental pollution in farmland. Endophytic nitrogen-fixing bacteria (ENFB) provide N nutrition for plants through biological N fixation, thus reducing the need for chemical fertilizers. The present study investigated the effect of the N-fixing endophytic strain Enterobacter roggenkampii ED5 on phytohormone indole-3-acetic acid (IAA), N-metabolism enzyme activities, microbial community compositions, and N cycle genes in sugarcane rhizosphere soil at different N levels. Three levels of 15N-urea, such as low N (0 kg/ha), medium N (150 kg/ha), and high N (300 kg/ha), were applied. The results showed that, after inoculating strain ED5, the IAA content in sugarcane leaves was significantly increased by 68.82% under low N condition at the seedling stage (60 days). The nitrate reductase (NR) activity showed a downward trend. However, the glutamine synthase (GS) and NADH-glutamate dehydrogenase (NADH-GDH) activities were significantly enhanced compared to the control under the high N condition, and the GS and NR genes had the highest expression at 180 and 120 days, respectively, at the low N level. The total N content in the roots, stems, and leaves of sugarcane was higher than the control. The 15N atom % excess of sugarcane decreased significantly under medium N condition, indicating that the medium N level was conducive to N fixation in strain ED5. Metagenome analysis of sugarcane rhizosphere soil exhibited that the abundance of N-metabolizing microbial richness was increased under low and high N conditions after inoculation of strain ED5 at the genus level, while it was increased at the phylum level only under the low N condition. The LefSe (LDA > 2, p < 0.05) found that the N-metabolism-related differential microorganisms under the high N condition were higher than those under medium and low N conditions. It was also shown that the abundance of nifDHK genes was significantly increased after inoculation of ED5 at the medium N level, and other N cycle genes had high abundance at the high N level after inoculation of strain ED5. The results of this study provided a scientific reference for N fertilization in actual sugarcane production.

Genomic and biotechnological potential of a novel oil-degrading strain Enterobacter kobei DH7 isolated from petroleum-contaminated soil

In this study, a novel oil-degrading strain Enterobacter kobei DH7 was isolated from petroleum-contaminated soil samples from the industrial park in Taolin Town, Lianyungang, China. The whole genome of the strain was sequenced and analyzed to reveal its genomic potential. The oil degradation and growth conditions including nitrogen, and phosphorus sources, degradation cycle, biological dosing, pH, and oil concentration were optimized to exploit its commercial application. The genome of the DH7 strain contains 4,705,032 bp with GC content of 54.95% and 4653 genes. The genome analysis revealed that there are several metabolic pathways and enzyme-encoding genes related to oil degradation in the DH7 genome, such as the paa gene cluster which is involved in the phenylacetic acid degradation pathway, and complete degradation pathways for fatty acid and benzoate, genes related to chlorinated alkanes and olefins degradation pathway including adhP, frmA, and adhE, etc. The strain DH7 under the optimized conditions has demonstrated a maximum degradation efficiency of 84.6% after 14 days of treatment using synthetic oil, which comparatively displays a higher oil degradation efficiency than any Enterobacter species known to date. To the best of our knowledge, this study presents the first-ever genomic studies related to the oil degradation potential of any Enterobacter species. As Enterobacter kobei DH7 has demonstrated significant oil degradation potential, it is one of the good candidates for application in the bioremediation of oil-contaminated environments.

The distribution of fitness effects of plasmid pOXA-48 in clinical enterobacteria.

Antimicrobial resistance (AMR) in bacteria is a major public health problem. The main route for AMR acquisition in clinically important bacteria is the horizontal transfer of plasmids carrying resistance genes. AMR plasmids allow bacteria to survive antibiotics, but they also entail physiological alterations in the host cell. Multiple studies over the last few years have indicated that these alterations can translate into a fitness cost when antibiotics are absent. However, due to technical limitations, most of these studies are based on analysing new associations between plasmids and bacteria generated in vitro, and we know very little about the effects of plasmids in their native bacterial hosts. In this study, we used a CRISPR-Cas9-tool to selectively cure plasmids from clinical enterobacteria to overcome this limitation. Using this approach, we were able to study the fitness effects of the carbapenem resistance plasmid pOXA-48 in 35 pOXA-48-carrying isolates recovered from hospitalized patients. Our results revealed that pOXA-48 produces variable effects across the collection of wild-type enterobacterial strains naturally carrying the plasmid, ranging from fitness costs to fitness benefits. Importantly, the plasmid was only associated with a significant fitness reduction in four out of 35 clones, and produced no significant changes in fitness in the great majority of isolates. Our results suggest that plasmids produce neutral fitness effects in most native bacterial hosts, helping to explain the great prevalence of plasmids in natural microbial communities.

Isolation of bacteria with plant growth-promoting properties from microalgae-bacterial flocs produced in high-rate oxidation ponds

ABSTRACT Exploring plant growth-promoting (PGP) bacterial activity of microbial components aggregated by wastewater treatment can reduce dependence on fossil fuel-derived fertilisers. This study describes the isolation and identification of bacteria from microalgae-bacteria flocs (MaB-flocs) generated in high-rate algal oxidation ponds (HRAOP) of an integrated algal pond system (IAPS) remediating municipal wastewater. Amplified 16S rRNA gene sequence analysis determined the molecular identity of the individual strains. Genetic relatedness to known PGP rhizobacteria in the NCBI GenBank database was by metagenomics. Isolated strains were screened for the production of indoles (measured as indole-3-acetic acid; IAA) and an ability to mineralise NH 4 + , PO 4 3 − , and K + . Of the twelve bacterial strains isolated from HRAOP MaB-flocs, four produced indoles, nine mineralised NH 4 + , seven solubilised P, and one K. Potential of isolated strains for PGP activity according to one-way ANOVA on ranks was: ECCN 7b > ECCN 4b > ECCN 6b > ECCN 3b = ECCN 10b > ECCN 1b = ECCN 5b > ECCN 8b > ECCN 2b > ECCN 12b > ECCN 9b = ECCN 11b. Further study revealed that cell-free filtrate from indole-producing cultures of Aeromonas strain ECCN 4b, Enterobacter strain ECCN 7b, and Arthrobacter strain ECCN 6b promoted mung bean adventitious root formation suggestive of the presence of auxin-like biological activity.

Promoting the growth of Sulla flexuosa L. by endophytic root nodule bacteria authors and affiliations.

Legume plants rely upon multipartite interactions between rhizobia and bacterial endophytes within root nodules to facilitate plant growth. This study aimed to isolate and identify indigenous endophytic bacteria from root nodules of Sulla aculeolata L. in Northeast Morocco. Based on their tri-calcium phosphate (TCP) solubilization capacity, five endophytes were chosen for further evaluation of their plant growth traits. All isolates were hydrogen cyanide (HCN) and siderophore producers, while only BCH24 tested positive for ACC deaminase activity. Indole-3-acetic acid (IAA) synthesis ranged from 1.27 mgL- 1 to 2.89 mgL- 1, while soluble phosphate concentrations was between 7.99mg L- 1 and 110.58mg L- 1. Additionally, all the endophytes were able to produce more than two lytic enzymes. Based on the analysis of 16S rRNA gene sequences five isolates were identified as Enterobacter sp (BCH13, BCH2), Pseudomonas sp (BCH16, BCH24), and Serratia sp (BCH10). The strains inhibited the growth of three phytopathogenic fungi, with BCH13 exhibiting the highest rate against Aspergillus ochraceus (45%), followed by BCH24 against Fusarium oxysporum (40%) and Botrytis cinerea (35%), respectively. In vivo inoculation of halotolerant strains Enterobacter hormaechei (BCH13) and Pseudomonas moraviensis (BCH16) under gnotobiotic conditions revealed that co-inoculation with Rhizobium sullae KS6 improved plant development compared to single inoculation, making it a promising eco-friendly bio-inoculant for legume Sulla flexuosa L. production.

Gut Bacteria of Snakeheads of Chinadi Beel, Narsingdi and Their Antibiogram Profile

The gut bacteria of snakehead fish (Channa punctata, C. marulius and C. striata) collected from Chinadi beel, Narsingdi, Bangladesh was characterized and formulated their antibiogram profile. The mean richness of intestinal bacteria in C. punctata was significantly higher in comparison to C. marulius and C. striata. A total of 24 isolates under the genus Bacillus, Enterobacter, Citrobacter, Kluyvera, Vibrio, Pseudomonas and Acinetobacter were identified. The strains of Enterobacter and Pseudomonas showed 100% resistance to eight antibiotics. The isolates of C. punctata were found to be resistant to nine antibiotics followed by C. marulius (8) and C. striata (7) indicating a critical condition of Chinadi beel of being polluted by anthropogenic activities. Bangladesh J. Bot. 52(2): 297-306, 2023 (June)

Molecular typing of multidrug resistant bacteria isolated from health care professionals’ mobile phone: A pilot study in Jashore, Bangladesh

Background: Nowadays, Health Care Professionals’ (HCPs) are increasingly using mobile phones which may act as reservoirs and vector for transmission of pathogens. The presence of multidrug resistant nosocomial microbes on the surface of mobile phones used by HCPs in hospitals can pose a great public health threat. So, this research was conducted to identify the concerned multi-drug resistant (MDR) bacteria and also to explore the recent status of bacterial contamination on mobile phones of HCPs in the Jashore region of Bangladesh and determine their antibiotic resistance pattern. Methods: Swab samples of mobile phones were collected between June and September 2019 from 24 different users (i.e., manager, worker, doctor and nurse) associated to four distinct hospitals of Jashore region, Bangladesh. After suitable morphological and biochemical identification, we determined their antimicrobial susceptibility by Kirby- Bauer disc diffusion method by using 18 antibiotics for Gram positive bacteria and 19 antibiotics for Gram negative bacteria. Later, the MDR isolates were grouped by amplified ribosomal DNA restriction analysis (ARDRA) and 16S rRNA sequencing with phylogeny were performed to confirm the bacteria at species level. Results: A total of 38 bacterial isolates were obtained from the sample. Enterobacter spp. isolates showed maximum resistance against Amoxicillin, followed by Ampicillin and Aztreonam (80% each) and one isolate showed highest antibiotic resistance (15 out of 19) among all the isolates. In addition, Staphylococcus spp. and Exiguobacterium spp. isolates showed 100% resistance against Penicillin, Ampicillin, Oxacillin, Erythromycin, Lincomycin and Cefotaxime. On the contrary, all of the isolates of Escherichia spp., Bacillus spp., Proteus spp. were sensitive to all tested antibiotics. Surprisingly, 20 MDR isolates were showing resistance to at least 2 antibiotics. Subsequently, three distinct genera of these MDR isolates were identified by ARDRA; the strains Enterobacter cloacae (75%), Staphylococcus warneri (15%) and Exiguobacterium aurantiacum (10%) were confirmed by the 16S rRNA phylogenetic analysis. Conclusion: We found that cell phones can act as reservoirs of multidrug-resistant pathogens, causative agents for Hospital-acquired infections. An effective hygiene practice for health care personnel should be introduced to prevent the cross-contamination by their cell phone Bangladesh Journal of Medical Science Vol. 22 No. 03 July’23 Page : 643-656

Isolation Characterization and Diversity of Indigenous Pesticide Degrading Microbes from Selected Agro Ecological Zones of Malawi

Pesticide xenobiotics have a great impact on bio argumentation, bio-magnification, and environment degradation regardless of being adopted by green revolution technologies. Bioremediation is widely accepted because it’s cheap, practical at the same time environmentally friendly. Bioremediation advocates indigenous microbes use to degrade pesticides, therefore a study has been performed to show prospects of degrading microorganisms. The study isolated microbes from different agro ecological zones to assess their capacity to utilize some pesticide as sole carbon source complimented by the presence of laccase gene. Biochemical test and genetic characterization using 16S rDNA genes were used in identification. Diversified species and strains of genus Enterobacter, Klebsiella, Pseudomonas, Pantoea and Leclercia, were found to degrade cypermethrin and acetochlor but no microbe was found to degrade dimethoate. The study adds new strain of microbes involved in degradation of cypermethrin and acetochlor and also strains that that can degrade both. The study puts proposition that pest infestation in fields is a result of abundance of xenobiotic degrading microbes due to natural selection pressure not pesticide resistance of the pest.

Citrobacter freundii resistant to novel β-lactamase inhibitor combinations and cefiderocol, co-producing class A, B and D carbapenemases encoded by transferable plasmids.

To characterize a carbapenem-resistant Citrobacter freundii (Cf-Emp) co-producing class A, B and D carbapenemases, resistant to novel β-lactamase inhibitor combinations (BLICs) and cefiderocol. Carbapenemase production was tested by an immunochromatography assay. Antibiotic susceptibility testing (AST) was performed by broth microdilution. WGS was performed using short- and long-read sequencing. Transfer of carbapenemase-encoding plasmids was assessed by conjugation experiments. Cf-Emp was isolated on selective medium for carbapenem-resistant Enterobacterales from the surveillance rectal swab taken at hospital admission from a patient of Moroccan origin. Cf-Emp produced three different carbapenemases, including KPC-2, OXA-181 and VIM-1, and was resistant to all β-lactams including carbapenems, novel BLICs (ceftazidime/avibactam, meropenem/vaborbactam and imipenem/relebactam) and cefiderocol. MIC of aztreonam/avibactam was 0.25 mg/L. The strain belonged to ST22, one of the C. freundii lineages of global diffusion, known to be associated with carbapenemase production. Each carbapenemase gene was located aboard a different plasmid (named pCf-KPC, pCf-OXA and pCf-VIM, respectively), which also carried other clinically relevant resistance genes, such as armA (pCf-KPC), blaSHV-12 (pCf-VIM) and qnrS1 (pCf-OXA). Transferability to Escherichia coli J53 by conjugation was observed for all plasmids. The finding of enterobacterial strains carrying multiple carbapenemase genes on transferable plasmids is alarming, because similar strains could provide an important reservoir for disseminating these clinically relevant resistance determinants.

Enterobacter pseudoroggenkampii sp. nov. carrying quinolone-resistant gene qnrE recovered from clinical samples in China.

Two Enterobacter strains 155092T and 170,225 were isolated from clinical samples, pus and sputum, from two hospitalised patients separately, in China. Preliminary identification using Vitek II microbiology system assigned the strains to the Enterobacter cloacae complex. The two strains were subjected to genome sequencing and genome-based taxonomy analysis with type strains of all Enterobacter species and those within closely related genera Huaxiibacter, Leclercia, Lelliottia, and Pseudoenterobacter. The average nucleotide identity (ANI) and in silico DNA-DNA hybridisation (isDDH) values between the two strains were 98.35% and 89.4%, respectively, suggesting that they belong to one species. The two strains had the highest ANI (95.02% and 95.04%) with the type strain of Enterobacter quasiroggenkampii. Their highest isDDH values, also seen with the type strain of E. quasiroggenkampii, were 59.5% and 59.8%, well below the 70% cutoff to define species. The two strains were also characterised for morphological and biochemical features by a set of experiments and observations. The abilities of metabolising gelatin and L-rhamnose could differentiate the two strains from all currently known Enterobacter species. Collectively, the two strains represent a novel Enterobacter species, for which we propose Enterobacter pseudoroggenkampii sp. nov. as the species name. The type strain of this novel species is155092T (= GDMCC 1.3415T = JCM 35646T). The two strains also carried multiple virulence factors comprising aerobactin-encoding iucABCD-iutA and salmochelin-encoding iroN. The two strains also had chromosomally located qnrE, a gene associated with reduced susceptibility to quinolones, suggesting that this species is a potential reservoir of qnrE genes.

Root exudate-derived compounds stimulate the phosphorus solubilizing ability of bacteria

Low phosphorus (P) availability in soils is a major challenge for sustainable food production, as most soil P is often unavailable for plant uptake and effective strategies to access this P are limited. Certain soil occurring bacteria and root exudate-derived compounds that release P are in combination promising tools to develop applications that increase phosphorus use efficiency in crops. Here, we studied the ability of root exudate compounds (galactinol, threonine, and 4-hydroxybutyric acid) induced under low P conditions to stimulate the ability of bacteria to solubilize P. Galactinol, threonine, and 4-hydroxybutyric acid were incubated with the P solubilizing bacterial strains Enterobacter cloacae, Pseudomonas pseudoalcaligenes, and Bacillus thuringiensis under either inorganic (calcium phosphate) or organic (phytin) forms of plant-unavailable P. Overall, we found that the addition of individual root exudate compounds did not support bacterial growth rates. However, root exudates supplemented to the different bacterial appeared to enhance P solubilizing activity and overall P availability. Threonine and 4-hydroxybutyric acid induced P solubilization in all three bacterial strains. Subsequent exogenous application of threonine to soils improved the root growth of corn, enhanced nitrogen and P concentrations in roots and increased available levels of potassium, calcium and magnesium in soils. Thus, it appears that threonine might promote the bacterial solubilization and plant-uptake of a variety of nutrients. Altogether, these findings expand on the function of exuded specialized compounds and propose alternative approaches to unlock existing phosphorus reservoirs of P in crop lands.

Unveiling the performance of a novel alkalizing bacterium Enterobacter sp. LYX-2 in immobilization of available Cd

A novel alkalizing strain Enterobacter sp. LYX-2 that could resist 400 mg/L Cd was isolated from Cd-contaminated soil, which immobilized 96.05% Cd2+ from medium. Cd distribution analysis demonstrated that more than half of the Cd2+ was converted into extracellular precipitated Cd through mobilization of the alkali-producing mechanism by the strain LYX-2, achieving the high immobilization efficiency of Cd2+. Biosorption experiments revealed that strain LYX-2 had superior biosorption capacity of 48.28 mg/g for Cd. Pot experiments with Brassica rapa L. were performed with and without strain LYX-2. Compared to control, 15.92% bioavailable Cd was converted to non-bioavailable Cd and Cd content in aboveground vegetables was decreased by 37.10% with addition of strain LYX-2. Available Cd was mainly immobilized through extracellular precipitation, cell-surface biosorption and intracellular accumulation of strain LYX-2, which was investigated through Cd distribution, Scanning Electron Microscope and Energy-Dispersive X-ray Spectroscopy (SEM-EDS), Fourier Transform Infrared Spectroscopy (FT-IR), X-ray Photoelectron Spectroscopy (XPS) and Transmission Electron Microscopy (TEM) analysis. In addition, the application of strain LYX-2 significantly promoted the growth of vegetables about 2.4-fold. Above results indicated that highly Cd-resistant alkalizing strain LYX-2, as a novel microbial passivator, had excellent ability and reuse value to achieve the remediation of Cd-contaminated soil coupled with safe production of vegetables simultaneously.

Chromate Removal by Enterobacter cloacae Strain UT25 from Tannery Effluent and Its Potential Role in Cr (VI) Remediation.

An indigenous chromate-resistant bacterial strain isolated from tannery effluent was identified based on morphological, biochemical, and 16S rRNA gene sequencing, as Enterobacter cloacae UT25. It was found to resist heavy metal ions such as Cr (VI), Pb (II), Cu (II), Co (II), Ni (II), Hg (II), and Zn (II) and antibiotics. The strain was able to remove 89 and 86% chromate, after 24h of incubation in a Luria-Bertani (LB) medium at an initial Cr (VI) concentration of 1000 and 1500µg/ml, respectively. Minimum inhibitory concentration (MIC) was observed for chromate to be 80,000 and 1850µg/ml, after 48h of incubation in LB and acetate minimal media (AMM), respectively. Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) analysis showed discrete cells with intact and smooth cell walls and homogenous cytoplasm in the absence of metal stress, whereas chromate stress caused cell lysis and reduction in size, which was a characteristic response to Cr (VI) toxicity. Energy Dispersive X-Ray Spectroscopy (EDX) confirmed the adsorption of oxyanions to the cell wall which was one of the Cr (VI) removal mechanisms by the bacterium. Atomic Force Microscopy (AFM) micrographs of chromate-untreated and treated cells revealed Root Mean Square roughness (Rq) values of 16.25 and 11.26nm, respectively, indicating less roughness in the presence of stress. The partial gene sequence of class 1 integrons (intI1) of strain UT25 showed 94% homology with intI1 gene of strain Enterobacter hormaechei strain ECC59 plasmid pECC59-1. The present analysis highlighted the potential of E. cloacae UT25 as a promissory bacterium that could be applied in removing chromate from polluted environments.

Profiling the composition and metabolic functions of microbial community in pellicle-forming radish paocai

Pellicle formation is an obvious indicator of spoilage and is followed by a loss of flavor in a variety of fermented vegetables. In this study, the pellicle-forming microorganisms were isolated using culture-dependent approaches, then a comparative analysis between the pellicle-forming (PF) radish paocai and normal fermented paocai in the diversity and function of microbial community was conducted by metagenome sequencing. Based on a pairwise t-test and OPLS-DA analysis, diallyl sulfide, (z)-1-allyl-2-(prop-1-en-1-yl) disulfane, and terpineol were considered to be the main components responsible for the unpleasant flavor of PF paocai. Yarrowia spp., Enterobacter spp., and Pichia spp. were the main pellicle-forming microorganisms. All 17 isolated Enterobacter strains showed pectinase-producing and cellulase-producing abilities, and 3 isolated Pichia strains showed gas-producing capacity. According to LEfSe analysis based on metagenomes, unclassified_g__Citrobacter and Yarrowia lipolytica were the uppermost biomarkers that distinguished the PF paocai from normal paocai. Unclassified_g__Lactobacillus and Lactobacillus plantarum were found to be actively engaged in starch and sucrose metabolism, cysteine and methionine metabolism, galactose metabolism, fructose and mannose metabolism, lysine biosynthesis, fatty acid biosynthesis, and arginine biosynthesis, all of which contributed to the flavor formation of paocai. Combining the results of metagenome sequencing with the data obtained based on the culture-dependent method, we could deduce that the growth of Yarrowia lipolytica first promoted the increase of pH and the formation of pellicle, which provided a suitable niche for the growth of some harmful bacteria such as Enterobacter, Citrobacter, and Serratia. These hazardous bacteria then worked in concert to induce the odorous stench and texture softening of paocai, as well as more pellicle formation.

Inactivation of Cd and As by an Enterobacter Isolated from Cd and As Contaminated Farmland Soil

A strain of Enterobacter was screened from cadmium and arsenic contaminated farmland soil and its passivation mechanism of cadmium and arsenic were explored through removing performance and characterization experiments. The results showed that the screened strain M5 was identified as Enterobacter sp. with a sulfate-reduction function, and its maximum resistance concentration was approximately 1 mmol·L-1 to cadmium and arsenic. In the simulation system, the maximum removal efficiencies of cadmium and arsenic were 94.13% and 27.26% by strain M5, respectively. The results of SEM-EDS and XRD confirmed that Cd and As were fixed to CdS and As2S3, and XPS results showed that carboxyl groups, hydroxyl groups, and amide groups on the surface of the bacteria were mainly involved in biological adsorption. These results can provide new ideas and a theoretical basis for microbial applications to soil remediations for heavy metal pollution.

Sustainable biodegradation of textile dye reactive blue 222 by the novel strain Enterobacter CU2004, isolated from the industrial waste: A design of experiment based optimization study and characterisation of metabolites

Sustainable biodegradation of textile dye reactive blue 222 by the novel strain Enterobacter CU2004, isolated from the industrial waste: A design of experiment based optimization study and characterisation of metabolites

First Detection of Fluoroquinolones and Aminoglycosides Resistance Genes in Multiresistant &lt;i&gt;Enterobacteriaceae&lt;/i&gt; from Urinary Tract Infections

Aminoglycosides and fluoroquinolones are among the most used antibiotics in synergy with beta-lactams. The objective of this work was to detect genes associated with resistance to fluoroquinolones and aminoglycosides in multiresistant strains of enterobacteria responsible for urinary tract infections. The diffusion method was used for antimicrobial sensitivity test of strains. qnr and aac genes were detected by Polymerase Chain Reaction. A total of 128 enterobacterial strains were studied. Resistance to the antibiotic families tested was observed with a frequency greater than 50%. Aminoglycosides were the least active family among the antibiotics used, with a resistance rate of 69.53%. aac(6’)-Ib-cr and qnrS genes were detected at rates of 34.09% and 22.72%, respectively. 13.63% of the strains showed coexpression of the qnr and aac genes. The detection of resistance genes to fluoroquinolones and aminoglycosides in enterobacterial strains isolated at the National Public Health Laboratory in Brazzaville could constitute a major concern in the therapeutic management of patients because it could lead to high costs for prolonged hospitalization.

Modification of root architecture of rice by guard cell and rhizosphere halotolerant bacteria under saline stress

A significant global issue for agriculture is soil salinity. Rice, being one of the most salt-sensitive cereals, is greatly affected by salinity. High salt concentrations have a detrimental impact on root growth of rice. Halotolerant bacteria are known to alleviate salinity stress and have been shown to alter the root morphology of some plants. This possible alteration effect of halotolerant bacteria on rice is not well known. In this study, the halotolerant bacterial strains Enterobacter cloacae GCH3, Acinetobacter sp. GCH4, Acinetobacter calcoaceticus RSH6 and Bacillus tequilensis RSH8 were used to evaluate the early alterations of rice root architecture. These bacteria were inoculated into rice seedlings growing in culture tubes under gnotobiotic conditions, and root architecture variables of developing roots were assessed periodically using non-destructive 2-dimensional imaging and processing in both normal and salt stress conditions. The images were analyzed and processed to obtain 20 root architecture variables in which, the variables such as maximum number of roots, network solidity, number of lateral roots, network surface area, root thickness, area, volume and bushiness were significantly increased with time course due to halotolerant bacterial inoculation than uninoculated control. The root and shoot biomass was also found to be increased by bacterial inoculation. Number of connecting components, median number of roots, specific root length and major ellipse axes were negatively impacted by halotolerant bacteria. Enterobacter cloacae GCH3 and Bacillus tequilensis RSH8 excelled in most of the traits in improving the root architecture of rice under saline conditions which can promote the early growth stages of root and protect the plant from saline stress effects.

Characterization and the host specificity of Pet-CM3–4, a new phage infecting Cronobacter and Enterobacter strains

Bacteria belonging to Cronobacter and Enterobacter genera are opportunistic pathogens responsible for infections in immunocompromised patients including neonates. Phage therapy offers a safe method for pathogen elimination, however, phages must be well characterized before application. In the present study we isolated four closely related bacteriophages from the subfamily Tevenvirinae infecting Cronobacter and Enterobacter strains. Bacteriophage Pet-CM3–4 which was isolated on C. malonaticus strain possessed broader host specificity than other three phages with primary Enterobacter hosts. Based on genome sequences all these phages have been assigned to the genus Karamvirus. We also studied factors influencing the host specificity of Pet-CM3–4 phage and its host range mutant Pet-CM3–1 and observed that a lysine to glutamine substitution in the long tail fiber adhesin was the reason of the Pet-CM3–1 reduced host specificity. By characterization of phage-resistant mutants from transposon library of C. malonaticus KMB-72 strain we identified that LPS is the receptor of both phages. C. malonaticus O:3 antigen is the receptor of Pet-CM3–1 phage and the Pet-CM3–4 phage binds to structures of the LPS core region. Obtained results will contribute to our understanding of biology and evolution of Tevenvirinae phages.

A comprehensive description of the TolC effect on the antimicrobial susceptibility profile in Enterobacter bugandensis.

Enterobacter bugandensis is an emerging human pathogen in which multidrug resistant strains have been continuously isolated from various environments. Thus, this organism possesses the potential to pose challenges in human healthcare. However, the mechanisms, especially the efflux pumps, responsible for the multidrug resistance in E. bugandensis remain to be well elucidated. The Enterobacter strain CMCC(B) 45301 was specifically identified using whole genome sequencing. The specific CMCC(B) 45301 homologues of the TolC dependent efflux-pump genes characterized in Escherichia coli were identified. The tolC deletion mutant in CMCC(B) 45301 was constructed and subjected to susceptibility tests using 26 different antimicrobial agents, along with the wild type strain. The synergistic effects combining the Bacillus crude extract (BCE) and several other TolC-affected compounds against CMCC(B) 45301 were assayed. We reclassified the Enterobacter CMCC(B) 45301 strain from species cloacae to bugandensis, on the basis of its whole genome sequence. We found that the CMCC(B) 45301 TolC, AcrAB, AcrD, AcrEF, MdtABC, EmrAB, and MacAB exhibit high similarity with their respective homologues in E. coli and Enterobacter cloacae. Our results for the susceptibility tests revealed that lacking tolC causes 4- to 256-fold decrease in the minimal inhibitory concentrations of piperacillin, gentamicin, kanamycin, tetracycline, norfloxacin, ciprofloxacin, chloramphenicol, and erythromycin against CMCC(B) 45301. In addition, the inhibition zones formed by cefuroxime, cefoperazone, amikacin, streptomycin, minocycline, doxycycline, levofloxacin, florfenicol, trimethoprim-sulfamethoxazole, azithromycin, lincomycin, and clindamycin for the tolC mutant were larger or more obvious than that for the parent. Our data suggested the important role played by TolC in CMCC(B) 45301 susceptibility to common antibiotic families covering ß-lactam, aminoglycoside, tetracycline, fluoroquinolone, phenicol, folate pathway antagonist, macrolide, and lincosamide. Deletion for tolC also increased the susceptibility of CMCC(B) 45301 to berberine hydrochloride and BCE, two natural product-based agents. Finally, we found that erythromycin, norfloxacin, and ciprofloxacin can potentiate the antibacterial activity of BCE against CMCC(B) 45301. The present study elaborated the comprehensive TolC effect on the antimicrobial susceptibility profile in E. bugandensis, which might contribute to the development of more therapeutic options against this nosocomial pathogen.