Enterobacter Cloacae Strain Research Articles

Newly isolated Enterobacter cloacae sp. HN01 and Klebsiella pneumoniae sp. HN02 collaborate with self-secreted biosurfactant to improve solubility and bioavailability for the biodegradation of hydrophobic and toxic gaseous para-xylene.

The gas-liquid mass transfer rate of hydrophobic volatile organic compounds (VOCs) is the limiting step in a biological treatment system. The present study aimed to utilize self-producing biosurfactants to enhance the bioavailability of hydrophobic gaseous VOCs. Two novel gram-negative rod-shaped bacteria, Enterobacter cloacae strain HN01 and Klebsiella pneumoniae strain HN02 were successfully isolated from sewage sludge by using blood agar and methylene blue agar plates. The two strains can use para-xylene (PX), a hydrophobic VOC model, as the only carbon source for biosurfactant production. Both strains can produce glycolipid biosurfactants, as confirmed by the emulsification index, Nuclear magnetic resonance, and Fourier transform infrared spectroscopy. Results indicated that PX can be completely decomposed at an initial concentration of 15.50mgL-1, pH value of 7.0, and temperature of 30°C within 36h. The Yano model is suitable for the prediction of the growth kinetics of strains over the entire PX concentration range. Gas chromatography/mass spectrometry analysis indicated that PX was converted into four and four intermediates in the presence of the strains HN01 and HN02, respectively, and the possible mechanisms were proposed. The results can be used in purifying industrial hydrophobic gaseous VOCs and improving the bioavailability of VOCs with self-produced biosurfactants.

Comparative genome analyses uncovered the cadmium resistance mechanism of enterobacter cloacae.

Cadmium (Cd) can be transported into plants from polluted soils and may cause animal and human diseases through food chains, which requires the development of highly efficient methods for soil Cd remediation. Although we isolated an Enterobacter cloacae strain Cu6 with Cd resistance, this strain cannot be used for soil Cd remediation due to its lower resistance. Here, we domesticated Cu6 and obtained a highly Cd-resistant strain, LPY6, and found that this strain can attenuate the toxic effects of Cd on wheat seedling growth. We deciphered the high Cd-resistance mechanism of LPY6 by genome comparative and genetic analysis. Compared with Cu6, 75 genes were mutated in LPY6. Thirty-four of these genes were deleted, and 41 had single nucleotide polymorphisms (SNPs). Most of these mutated proteins are involved in basic metabolism, substrate transport, stress response and formate and hydrogen metabolism. RNA quantitative analysis and promoter activity assays showed that the transcription or mRNA levels of two operons (cadA and norVW) in these mutated genes were regulated by Cd, zinc (Zn) or lead (Pb) ions, suggesting that these two operons might be required for Cd, Zn or Pb resistance. Expression of cadA and norVW operons in LPY6 partially recovered Cd susceptibility, demonstrating that CadA and NorVW are involved in Cd resistance in E. cloacae. Our findings illustrate that E. cloacae acquires Cd resistance through different pathways and lay a foundation for developing highly efficient methods for soil Cd remediation.

Biofilm formation during wastewater treatment: Motility and physiological response of aerobic denitrifying bacteria under ammonia stress based on surface plasmon resonance imaging

A bacterial image analysis system based on surface plasmon resonance imaging was established to investigate the effect of bacterial motility on biofilm formation under high ammonia nitrogen at the single-cell level. The results showed that the bacterial mean rotation speed and vertical motility distance decreased with the increasing concentration of ammonia nitrogen. Ammonia nitrogen inhibited the metabolic activity of the bacteria, decreasing bacterial motility. Bacterial motility was negatively correlated with the biofilm-formation ability. The biofilm formation ability of Enterobacter cloacae strain HNR exposed to ammonia nitrogen was enhanced by reducing its movement and promoting EPS secretion. Genes related to the tricarboxylic acid cycle and oxidative phosphorylation were down-regulated, indicating inhibition of microbial energy metabolism. Genes related to bacterial secretion and lipopolysaccharide synthesis were up-regulated, facilitating the formation of biofilms and enabling the bacteria to resist ammonia nitrogen stress. This study provides new insights into the biofilm formation under ammonia stress.

Molecular Characterization of Enterobacter cloacae Isolated from Urinary Tract Infections

Background: Urinary tract infections represent a major expensive, common public health problem worldwide due to their high prevalence and the difficulties associated with their management. Objectives: This study aimed to characterize the Enterobacter cloacae strains isolated from urinary tract infections in the medical diagnostic laboratories of Shahrekord, Iran. Methods: Urine samples from patients with urinary tract infections from the Shahrekord medical diagnostic laboratories located in Chaharmahal and Bakhtiari Province, Iran, were collected from June 2019 to February 2020. When the samples were cultured, the different isolates of E. cloacae were identified by biochemical tests. Biofilm production capacity was evaluated. Bacterial susceptibility to antibiotics was determined using the Kirby Bauer method, and antibiotic resistance genes were researched by the multiplex PCR technique. Results: In this study, 65 isolates of E. cloacae were obtained. The highest percentage of resistance was observed for co-trimoxazole (84.62%), ampicillin (76.93%), tetracycline (73.85%), and above half of the E. cloacae strain isolates (53,85%) were strongly involved in biofilm production. Some genes, including qnr A, qnr B, qnr S, tetA, tet B, sul1, bla CTXM, bla SHV, and(2)la, ant(3)la, and aac(3)IIa, were detected in the genome of these isolates. Conclusions: The strains are multi-resistant, and their resistance has already reached the carbapenem class. This requires further investigation, and urgent measures must be adopted.

Efficiency of halophilic biofilm producing bacteria towards the degradation of plastic materials at optimum temperature

The consumption of plastic has drastically increased leads to the plastic waste and became the global issue. In the present study, the screening of bacterial isolates from saline areas along with their microbial and biofilm efficiency in degradation of low-density polyethylene (LDPE), high density polyethylene (HDPE) and polypropylene plastic materials were estimated at two different temperatures (30°C and 37°C). The soil samples were collected from salt-affected lands for the isolation and characterization of bacterial isolates. The isolated strains were characterized by 16S rRNA. Two bacterial strains (Bacillus subtilis and Enterobacter cloacae) were identified through sequencing (BioEditor Sequence Builder) among the selected bacterial isolates. Effective degradation rate has been observed through B. subtilis towards LDPE, HDPE and polypropylene as 18%, 25% and 42% respectively through biofilm, while the degradation rate in TSA media were observed as 32%, 30% and 52% respectively, at 37°C. Similarly, E. cloacae degrades the LDPE, HDPE and polypropylene material at 12%, 15% and 30% through biofilm, however 19%, 18% and 38% degradation rate were observed at 37°C respectively. Therefore, both bacterial strains (MK2 B. subtilis and MK29 E. cloacae) isolated from salt-affected area showed potential to degrade the plastic materials at optimum temperature of 37°C.

Heavy Metal-Resistant Plant Growth-Promoting Citrobacter werkmanii Strain WWN1 and Enterobacter cloacae Strain JWM6 Enhance Wheat (Triticum aestivum L.) Growth by Modulating Physiological Attributes and Some Key Antioxidants Under Multi-Metal Stress.

Due to wastewater irrigation, heavy metal (HM) exposure of agricultural soils is a major limiting factor for crop productivity. Plant growth–promoting bacteria (PGPB) may lower the risk of HM toxicity and increase crop yield. In this context, we evaluated two HM-resistant PGPB strains, i.e., Citrobacter werkmanii strain WWN1 and Enterobacter cloacae strain JWM6 isolated from wastewater-irrigated agricultural soils, for their efficacy to mitigate HM (Cd, Ni, and Pb) stress in a pot experiment. Increasing concentrations (0, 50, 100, and 200 ppm) of each HM were used to challenge wheat plants. Heavy metal stress negatively affected wheat growth, biomass, and physiology. The plants under elevated HM concentration accumulated significantly higher amounts of heavy metals (HMs) in shoots and roots, resulting in increased oxidative stress, which was evident from increased malondialdehyde (MDA) content in roots and shoots. Moreover, alterations in antioxidants like superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT) were observed in plants under HM stress. The severity of damage was more pronounced with rising HM concentration. However, inoculating wheat with Citrobacter werkmanii strain WWN1 and Enterobacter cloacae strain JWM6 (107 CFU ml–1) improved plant shoot length (11–42%), root length (19–125%), fresh weight (41–143%), dry weight (65–179%), and chlorophyll a (14%-24%) and chlorophyll b content (2–24%) under HM stress. Citrobacter werkmanii strain WWN1 and Enterobacter cloacae strain JWM6 either alone or in co-inoculation enhanced the antioxidant enzyme activity, which may lower oxidative stress in plants. However, seeds treated with the bacterial consortium showed an overall better outcome in altering oxidative stress and decreasing HM accumulation in wheat shoot and root tissues. Fourier transform infrared spectroscopy indicated the changes induced by HMs in functional groups on the biomass surface that display effective removal of HMs from aqueous medium using PGPB. Thus, the studied bacterial strains may have adequate fertilization and remediation potential for wheat cultivated in wastewater-irrigated soils. However, molecular investigation of mechanisms adopted by these bacteria to alleviate HM stress in wheat is required to be conducted.

Utilization of sugarcane molasses as the main carbon source for the production of polyhydroxyalkanoates from Enterobacter cloacae

Polyhydroxyalkanoates (PHAs) are biodegradable polymers that otherwise exhibit many properties similar to that of conventional, non-biodegradable plastics. Hence, they are considered as potential substitutes for non-biodegradable plastics. But industrial production and commercialization of PHA have been hindered greatly due to the high cost requirements, especially those involved in the provision of suitable carbon sources. In this study, sugarcane molasses was used as a cheap carbon source for PHA production using a wild strain of Enterobacter cloacae isolated from sugarcane extract. This isolate was selected for PHA production based on Sudan Black B staining, and it was subsequently identified by performing BLASTn using its 16S rRNA sequence (GenBank accession number: ON314993). The PHA yield was studied under varying conditions of initial pH, molasses concentration and inoculum concentration. The highest yield of 4.13 - 4.98 g/L or 48 - 56% PHA was obtained after 48-60 h incubation at initial pH 7, molasses concentration 4% and inoculum concentration 2%. After downstream processing, the extracted PHA was characterized by FTIR spectroscopy. It is concluded that E. cloacae has the potential to utilize sugarcane molasses as a cheap carbon source to yield impressive amounts of PHA, and that it should be considered a candidate for further studies and for the cheaper industrial production of PHA.

Single Center Clinical Analysis of Bloodstream Infection Pathogens in Children with Acute Leukemia

To investigate the clinical features, distribution of pathogenic bacteria, and drug resistance of bloodstream infection in children with acute leukemia. Clinical data of 93 blood culture-positive children with acute leukemia from January 2015 to December 2019 in Department of Pediatrics, The Second Hospital of Anhui Medical University were analyzed retrospectively. In these 93 cases, 78 cases were in the period of neutrophil deficiency. There were 54 Gram-negative bacteria (G-) (58.1%) found through blood culture, and the top 4 strains were Escherichia coli (15.1%), Klebsiella pneumoniae (13.9%), Pseudomonas aeruginosa (6.5%), and Enterobacter cloacae (6.5%). There were 39 Gram-positive bacteria (G+) (41.9%) detected, and the top 4 strains were Staphylococcus epidermidis (10.8%), Streptococcus pneumoniae (6.5%), Staphylococcus hemolyticus (5.4%), and Staphylococcus human (5.4%). Among 74 strains of pathogenic bacteria from acute lymphoblastic leukemia (ALL) children, there were 29 strains of G+ bacteria (39.2%) and 45 strains of G- bacteria (60.8%). While in 19 strains from acute myeloblastic leukemia (AML) patients, G- bacteria accounted for 47.4% and G+ bacteria accounted for 52.6%. In 15 ALL children without neutropenia, G+ bacteria made up the majority of the strains (66.7%). In the 93 strains of pathogenic bacteria, 13 (13.9%) strains were multidrug-resistant. Among them, extended-spectrum β-lactamases accounted for 42.9%, carbapenemase-resistant enzyme Klebsiella pneumoniae 15.4%, and carbapenemase-resistant enzyme Enterobacter cloacae strains 33.3%, which were detected from G- bacteria. While, 13.3% of methicillin-resistant coagulase-negative Staphylococci accounted for 13.3% detected from G+ bacteria, but linezolid, vancomycin, teicoplanin Staphylococcus and Enterococcus resistant were not found. The average procalcitonin (PCT) value of G- bacteria infection was (11.02±20.282) ng/ml, while in G+ infection it was (1.81±4.911) ng/ml, the difference was statistically significant (P<0.05). The mean value of C-reactive protein (CRP) in G- infection was (76.33±69.946) mg/L, and that in G+ infection was (38.34±57.951) mg/L. The prognosis of active treatment was good, and only one case died of septic shock complicated with disseminated intravascular coagulation (DIC) and gastrointestinal bleeding caused by carbapenemase-resistant enzyme enterobacteriaceae. G- is the major bacteria in acute leukemia children with bloodstream infection, but the distribution of ALL and AML strains is different. G- bacteria dominates in ALL, while G+ bacteria and G- bacteria are equally distributed in AML. Non-agranulocytosis accompanied by bloodstream infections is dominant by G+ bacteria. The mean value of PCT and CRP are significantly higher in G- bacteria infection than in G+ bacteria.

A Multiwell-Plate Caenorhabditis elegans Assay for Assessing the Therapeutic Potential of Bacteriophages against Clinical Pathogens.

ABSTRACTIn order to establish phage therapy as a standard clinical treatment for bacterial infections, testing of every phage to ensure the suitability and safety of the biological compound is required. While some issues have been addressed over recent years, standard and easy-to-use animal models to test phages are still rare. Testing of phages in highly suitable mammalian models such as mice is subjected to strict ethical regulations, while insect larvae such as the Galleria mellonella model suffer from batch-to-batch variations and require manual operator skills to inject bacteria, resulting in unreliable experimental outcomes. A much simpler model is the nematode Caenorhabditis elegans, which feeds on bacteria, a fast growing and easy to handle organism that can be used in high-throughput screening. In this study, two clinical bacterial strains of Escherichia coli, one Klebsiella pneumoniae, and one Enterobacter cloacae strain were tested on the model system together with lytic bacteriophages that we isolated previously. We developed a liquid-based assay, in which the efficiency of phage treatment was evaluated using a scoring system based on microscopy and counting of the nematodes, allowing increasing statistical significance compared to other assays such as larvae or mice. Our work demonstrates the potential to use Caenorhabditis elegans to test the virulence of strains of Klebsiella pneumoniae, Enterobacter cloacae, and EHEC/EPEC as well as the efficacy of bacteriophages to treat or prevent infections, allowing a more reliable evaluation for the clinical therapeutic potential of lytic phages.IMPORTANCE Validating the efficacy and safety of phages prior to clinical application is crucial to see phage therapy in practice. Current animal models include mice and insect larvae, which pose ethical or technical challenges. This study examined the use of the nematode model organism C. elegans as a quick, reliable, and simple alternative for testing phages. The data show that all the four tested bacteriophages can eliminate bacterial pathogens and protect the nematode from infections. Survival rates of the nematodes increased from <20% in the infection group to >90% in the phage treatment group. Even the nematodes with poly-microbial infections recovered during phage cocktail treatment. The use of C. elegans as a simple whole-animal infection model is a rapid and robust way to study the efficacy of phages before testing them on more complex model animals such as mice.

PGPR-Mediated Salt Tolerance in Maize by Modulating Plant Physiology, Antioxidant Defense, Compatible Solutes Accumulation and Bio-Surfactant Producing Genes.

Salinity stress is a barrier to crop production, quality yield, and sustainable agriculture. The current study investigated the plant growth promotion, biochemical and molecular characterization of bacterial strain Enterobacter cloacae PM23 under salinity stress (i.e., 0, 300, 600, and 900 mM). E. cloacae PM23 showed tolerance of up to 3 M NaCl when subjected to salinity stress. Antibiotic-resistant Iturin C (ItuC) and bio-surfactant-producing genes (sfp and srfAA) were amplified in E. cloacae PM23, indicating its multi-stress resistance potential under biotic and abiotic stresses. Moreover, the upregulation of stress-related genes (APX and SOD) helped to mitigate salinity stress and improved plant growth. Inoculation of E. cloacae PM23 enhanced plant growth, biomass, and photosynthetic pigments under salinity stress. Bacterial strain E. cloacae PM23 showed distinctive salinity tolerance and plant growth-promoting traits such as indole-3-acetic acid (IAA), siderophore, ACC deaminase, and exopolysaccharides production under salinity stress. To alleviate salinity stress, E. cloacae PM23 inoculation enhanced radical scavenging capacity, relative water content, soluble sugars, proteins, total phenolic, and flavonoid content in maize compared to uninoculated (control) plants. Moreover, elevated levels of antioxidant enzymes and osmoprotectants (Free amino acids, glycine betaine, and proline) were noticed in E. cloacae PM23 inoculated plants compared to control plants. The inoculation of E. cloacae PM23 significantly reduced oxidative stress markers under salinity stress. These findings suggest that multi-stress tolerant E. cloacae PM23 could enhance plant growth by mitigating salt stress and provide a baseline and ecofriendly approach to address salinity stress for sustainable agriculture.

Metabolic properties, gene functions, and biosafety analysis reveal the action of three rhizospheric plant growth-promoting bacteria of Jujuncao (Pennisetum giganteum).

This study aimed to identify the specific genes associated with plant growth promotion and cadmium tolerance in three bacteria strains associated with Pennisetum giganteum as well as to determine their biosafety levels in their potential use as biofertilizers for promoting plant growth and phytoremediation activities. The plant growth-promoting (PGP) abilities of Enterobacter cloacae strain RCB980 (A3), Klebsiella pneumonia strain kpa (A4), and Klebsiella sp. strain XT-2 (A7) were determined by a growth promotion trial and through testing for PGP traits such as 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase enzyme production, phosphorus solubilization, siderophore synthesis, and indole-3 acetic acid (IAA) production. The genes that potentially contribute to the beneficial activities of these three strains were identified through an analysis of their genomes. To establish the biosafety of the candidate PGPB, a pathological study was undertaken whereby 20 Kunming mice were injected intraperitoneally to study and analyze the effects of the strains on growth and lung paraffin sections of the mice. The strains had no obvious toxicity effect on the tested mice and were therefore not considered as highly virulent strains. These strains are thus considered non-toxic, safe, and highly recommended for use in environmental remediation strategies and agricultural production.

Characterization of TEM and CMY-2 beta-lactamases in Enterobacteriaceae strains isolated at the Brazzaville University and hospital center in Congo

This study aimed to characterize the genes coding for ESBLs in TEM and CMY-2 Enterobacteriaceae. 65 strains of Enterobacteriaceae were isolated from 65 patients, of which 32.43% came from outpatients and 67.56% from hospitalized patients. The study was carried out over 6 months. The samples (urine, pus, and blood) were collected from hospitalized and external patients at the Brazzaville University Hospital. Strains were identified by API20E. An antibiogram was by diffusion in agar medium (MH) according to CA-SFM. The ESBL phenotype was detected by synergy. The plasmid cephalosporinase phenotype was determined by decreasing the diameter of inhibition around the cefoxitime disc in a strain resistant to ESBL inhibitors. Antibiotic resistance genes were characterized by PCR and sequencing. Sequences were assembled by Codon Code Aligner and compared by BlastX with Genbank, NCBI, and ARGANNOT. Phylogenetic gene trees were constructed using MEGA 7 software. 65 strains of Enterobacteriaceae were isolated from 65 patients, of which 32.43% came from outpatients and 67.56% from hospitalized patients. Except for imipenem, colistin; the amikacin and fosfomycin, tested antibiotics show high resistance to most of the beta-lactam, as well as resistance to very frequent aminoglycosides, sulfamides, tetracyclines, and Fluoroquinolones. PCR reveals that 97.29% possessed the TEM gene and 5.40% produced the CMY-2 gene. 64% of the isolates were isolated from urine samples, with a predominance of Enterobacter cloacae strains on ESBL production (75%). 60% of ESBL strains were isolated in the service of metabolic diseases. Sequencing of the amplification products revealed that the blaCTX-M1 strains were all TEM-1. Une consecvation des motifs SxxK en positions ABL 71 à 74 était observée pour l’alignement des sequences du groupe TEM. This study revealed the presence of the TEM and CMY-2 genes at disturbing frequencies. It proves the need to promote an infection prevention program with the regulation of antibiotic therapy in hospitals in Congo Brazzaville.

Infection Control for a Carbapenem-Resistant Enterobacteriaceae Outbreak in an Advanced Emergency Medical Services Center

Background: A carbapenem-resistant Enterobacteriaceae (CRE) outbreak occurred in an advanced emergency medical service center [hereafter referred to as the intensive care unit (ICU)] between 2016 and 2017. Aim: Our objective was to evaluate the infection control measures for CRE outbreaks. Methods: CRE strains were detected in 16 inpatients located at multiple sites. Environmental cultures were performed and CRE strains were detected in 3 of 38 sites tested. Pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and detection of β-lactamase genes were performed against 25 CRE strains. Findings: Molecular typing showed the PFGE patterns of two of four Klebsiella pneumoniae strains were closely related and the same MLST (ST2388), and four of five Enterobacter cloacae strains were closely related and same MLST (ST252). Twenty-three of 25 CRE strains harbored the IMP-1 β-lactamase gene and 15 of 23 CRE strains possessed IncFIIA replicon regions. Despite interventions by the infection control team, new inpatients with the CRE strain continued to appear. Therefore, the ICU was partially closed and the inpatients with CRE were isolated, and the ICU staff was divided into two groups between inpatients with CRE and non-CRE strains to avoid cross-contamination. Although the occurrence of new cases dissipated quickly after the partial closure, a few months were required to eradicate the CRE outbreak. Conclusion: Our data suggest that the various and combined measures that were used for infection control were essential in stopping this CRE outbreak. In particular, partial closure to isolate the ICU and division of the ICU staff were effective.

Molecular Characterization and Toxicity of Heavy Metal Quinary Mixtures on Enterobacter cloacae Isolate from Nworie River

The molecular characterization and toxicity of Cadmium (Cd), Lead (Pb), Zinc (Zn), Chromium (Cr) and Copper (Cu) quinary mixtures on Enterobacter cloacae isolate from Nworie River was investigated. Quinary mixtures of the heavy metals were compounded using fixed ratio (%); and inhibitory effect assessed using inhibition of total dehydrogenase as toxicity response. The mixtures consisted of five heavy metals in the ratios: Pb (20%) + Cr (20%) + Cd (20%) + Zn (20%) + Cu (20%), Pb (30%) + Cr (20%) + Cd (10%) + Zn (30%) + Cu (10%). Pb (10%) + Cr (10%) + Cd (30%) + Zn (40%) + Cu (10%), Pb (15%) + Cr (25%) + Cd (25%) + Zn (15%) + Cu (20%), Pb (40%) + Cr (15%) + Cd (5%) + Zn (20%) + Cu(20%). Result obtained showed that toxicity of the metals against the bacterium ranked in the order Cu &gt; Cr &gt;Zn &gt; Pb &gt; Cd. The quinary mixtures Pb (20%) + Cr (20%) + Cd (20%) + Zn (20%) + Cu (20%), Pb (30%) + Cr (20%) + Cd (10%) + Zn (30%) + Cu (10%) and 10%Pb + 10%Cr + 30%Cd + 40%Zn + 10%Cu showed synergistic interaction, while 15%Pb + 25%Cr + 25%Cd + 15%Zn + 20%Cu and 40%Pb + 15%Cr + 5%Cd + 20%Zn + 20%Cu mixtures were antagonistic and additive respectively. Threshold inhibitory concentration (IC50) of the quinary mixtures was 0.054 ± 0.005mM, 0.053 ± 0.002 mM, 0.038 ± 0.002 mM, 0.077 ± 0.007 mM and 0.058 ± 0.006 mM for the respective mixtures. The toxic index values evaluated for the mixtures exhibited antagonistic, synergistic and additive interaction in the various ratios. The 16S rRNA and ITSF1 sequences obtained of the isolate showed the phylogenetic placement of the 16S rRNA of the isolate was closely related to Enterobacter cloacae strain DL01 (MH168084). The heavy metals mixtures demonstrated diverse toxicity interaction on the isolate depending on their relative composition, thus poses a threat to aquatic microbial diversity.

Validation of Modifications to the Soleris®E. coli Method for Detection and Threshold Determination of Escherichia coli in Select Foods: Level 3 Modification to AOAC Performance Tested MethodSM 101101.

Soleris®E. coli is an automated, growth-based method for detection and semi-quantitative determination of Escherichia coli in foods. The method can be used in dilute-to-specification (threshold) or presence/absence modes. The objective of the study was to validate four modifications to the method: (1) a change in the vial detection window plug composition from agar to agarose to improve plug consistency and robustness, (2) a change in pre-enrichment incubation time for presence/absence testing from 6 h to 18-24 h, (3) a change in vial incubation temperature from 44.5 to 43.5°C, and (4) incorporation of a simple direct-from-vial confirmation test as an alternative to traditional procedures. Elements of the study included inclusivity/exclusivity testing, matrix testing in comparison to the ISO 7251:2005 reference method, reagent stability/lot-to-lot consistency testing, and an independent laboratory study. In inclusivity testing, all 55 Escherichia coli strains tested produced positive results. In exclusivity testing, 30 of 31 strains of other bacterial species produced negative results, the sole exception being a strain of Enterobacter cloacae. In internal and independent laboratory matrix testing of mozzarella cheese, condensed milk, pasteurized liquid egg, and frozen green beans, results showed no significant differences in performance of the Soleris and reference methods with two exceptions, one in which the Soleris method produced more positive results, and one in which the reference method produced more positive results. Performance characteristics of the modified Soleris E. coli method are consistent with those of the original validated method. The Soleris E. coli method offers improvements in ease of use and reagent robustness.

Selenium-enriched Polysaccharide: an Effective and Safe Selenium Source of C57 Mice to Improve Growth Performance, Regulate Selenium Deposition, and Promote Antioxidant Capacity.

Selenium-enriched polysaccharide (SeEPS) was prepared by reducing Se(IV) to elemental selenium and organic selenium in polysaccharide medium by the obtained Enterobacter cloacae strain Z0206 under aerobic conditions. In the present study, we focused on investigating the role of short-term supplementation of SeEPS at supernutritional doses in the regulation of growth performance, liver damage, antioxidant capacity, and selenium (Se) accumulation in C57 mice. Thirty-two C57 mice were randomly divided into four groups: the control group was gavaged with equal volume of phosphate-buffered saline, while the sodium selenite (Na2SeO3), selenomethionine (SeMet), and SeEPS groups were gavaged with 0.5mg Se/kg BW of Na2SeO3, SeMet, and selenium-enriched polysaccharide (n = 8), respectively. We examined liver injury indicators, antioxidant capacity in the serum and liver, selenium deposition at different sites, selenoprotein levels, and selenocysteine-synthesizing and degradation-associated gene expression in mouse livers. SeEPS supplementation dramatically increased average daily weight gain but reduced the feed-to-gain ratio (F/G) of mice (P < 0.05). Compared to Na2SeO3 and SeMet supplementation, SeEPS supplementation at supernutritional doses did not cause the liver damage. SeEPS supplementation also markedly enhanced total antioxidant capacity (T-AOC), catalase (CAT), glutathione peroxidase (GSH-PX), and total superoxide dismutase (T-SOD) activities but reduced malondialdehyde (MDA) levels in the liver and serum (P < 0.05), while significantly increasing selenocysteine-synthesizing and degradation-related gene (SEPHS2, SEPSECS, Secisbp, Scly) expression at the mRNA level (P < 0.05), thus upregulating the mRNA levels of selenoproteins (SELENOP, SELENOK) (P < 0.05). We suggest that SeEPS could be a potential replacement for inorganic selenium to improve animals' growth performance, promote antioxidant capacity, and regulate selenium deposition.

Biohydrogen Production from Aquatic Plant and Algae Biomass by Enterobacter cloacae Strain DT‐1

AbstractProcess parameters were optimized for enzyme (textile‐grade enzymes) hydrolysis of Azolla, water hyacinth, and algae biomass for recovery of maximum sugar to be used as feed for biohydrogen production by Enterobacter cloacae strain DT‐1. Under optimum temperature and pH, the hydrogen yield in mole per mole of sugar from enzyme‐hydrolyzed water hyacinth biomass, Azolla biomass, and Spirulina and Scenedesmus biomass was determined with regard to cellulase, hemicellulase, amylase, pectinase, and xylanase. The maximum hydrogen yield was observed with hemicellulase‐treated Spirulina, Azolla, and Scenedesmus biomass.

Isolation and identification of high fluoride resistant bacteria from water samples of Dindigul district, Tamil Nadu, South India.

Fluoride (F - ) pollution is one of the major issues in India and worldwide. Water samples were collected, and analyse the physicochemical characteristics. From the results, acceptable limit pH ranges, low and high electrical conductivity (EC) values, high-level of TDS (total dissolved solids) and chloride (Cl-) values, less than desirable and higher than permissible F - concentrations (4.7 and 11 ppm) were found. At first, ninety-three colonies were screened using rapid hicoliform agar plates. After that, sixty-six F - resistant colonies were picked up from 50 mM NaF (Sodium fluoride) containing LB agar plates. Finally, eight isolates were showing a high degree of F - resistance (200--300mM NaF) and selected for further studies. High F - resistance isolates exhibited β and γ-haemolytic activities were determined in blood agar plates. F - resistant isolates showed their salt tolerances ranged from 4% to 7% NaCl and resistant to multiple antibiotics. Biochemical and 16S rRNA sequencing results showed that the F - resistant isolates were identified as Enterobacter cloacae strain 3, E. hormaechei strain 14, Enterobacter sp. strain 21, E. hormaechei strain 22, E. coli strain S2-9, Aeromonas caviae strain 31, A. caviae strain 32, A. caviae strain 34. All strains were submitted in the NCBI database with the accession numbers (MW131637, MW131639; MW131650-655). The F - resistant gene 'crcB' gene was successfully amplified from the resistant isolates using gene-specific primers. These results have demonstrated that fluoride resistant bacteria would be useful for bacterial fluoride bioremediation near future.

Identification of Enterobacteriaceae members and fluorescent pseudomonads associated with bacterial rind necrosis and rot of melon in Turkey

During the 2016 growing season, melon fruits were severely affected by rind necrosis and rot and nearly 80% of the fruits were unmarketable in a field in Kocarli district of Aydin. In order to determine occurrences of the fruit infections in the following year, surveys were conducted in 2017. Isolations were made from the symptomatic fruits and bacterial growths were observed consistently. In this study, we identified the causal agents of melon fruit necrosis and rot. Pathogenicity of the bacterial strains was determined on melon fruit by needle injection method and thirteen strains were further characterized with phenotypic characterization, analysis of 16S rRNA gene sequences and specific PCR assays. As a result of the pathogenicity tests, brown spots and necrosis formation was observed initially and in the later stage, inoculated fruit burst out by the fermentative strains. The oxidative strains produced limited necrosis on the rind. Fermentative bacterial agents isolated from the melon necrosis and rot were identified as Enterobacter cloacae, Klebsiella pneumoniae and Kosakonia cowanii. Enterobacter cloacae strains were further tested with tDNA-PCR using T5A/T3B primers. The oxidative bacteria were phenotypically grouped within the fluorescent pseudomonads. Homology search of the oxidative strains using the sequence information of partial 16S rRNA gene identified the strains as Pseudomonas spp. The oxidative strain 413-Si2-L2 and the reisolate 06R2K were the most similar to strains of Pseudomonas putida and P. reidholzensis including the type strains. The strain, 413-MKV-3 and the reisolate 03R2K were the most related to Pseudomonas fluorescens, P. baetica, P. syringae, and Pseudomonas sp. strains. The ability of Enterobacter cloacae, Klebsiella pneumoniae and Kosakonia cowanii to cause disease on melon fruit was shown for the first time in this study by fulfilling Koch postulate. Furthermore, although limited rind necrosis was produced, pathogenicity of the fluorescent pseudomonads on melon fruit was also confirmed.

Adaptation mechanism of aerobic denitrifier Enterobacter cloacae strain HNR to short-term ZnO nanoparticle stresses

The adaptation mechanism of a wild type (WT) and resistant type (Re) strain of the aerobic denitrifier Enterobacter cloacae strain HNR to short-term ZnO nanoparticle (NP) stresses was investigated. The results showed that Re maintained higher nitrite reductase (NIR) and nitrate reductase (NR) activities and showed lower increment of reactive oxygen species (ROS) than WT, under ZnO NP stresses. The affinity constant (KA) of WT to Zn2+ was 5.06 times that of Re, indicating that Re was more repulsive to Zn2+ released by ZnO NPs. Transcriptomic analysis revealed that the up-regulation of the nitrogen metabolism of Re helped maintain NIR and NR activities, that the enhancement of purine metabolism lowered the intracellular ROS increment, and that the up-regulation of cationic antimicrobial peptide resistance contributed to the lower KA of Re to Zn2+. These findings provided new insights into the adaptation mechanism of aerobic denitrifying bacteria to ZnO NPs.