Cloacae Strain Research Articles

Expression of Zinc Transporter Genes in Rice as Influenced by Zinc-Solubilizing Enterobacter cloacae Strain ZSB14.

Zinc (Zn) deficiency in major food crops has been considered as an important factor affecting the crop production and subsequently the human health. Rice (Oryza sativa) is sensitive to Zn deficiency and thereby causes malnutrition to most of the rice-eating Asian populations. Application of zinc solubilizing bacteria (ZSB) could be a sustainable agronomic approach to increase the soil available Zn which can mitigate the yield loss and consequently the nutritional quality of rice. Understanding the molecular interactions between rice and unexplored ZSB is useful for overcoming Zn deficiency problems. In the present study, the role of zinc solubilizing bacterial strain Enterobacter cloacae strain ZSB14 on regulation of Zn-regulated transporters and iron (Fe)-regulated transporter-like protein (ZIP) genes in rice under iron sufficient and deficient conditions was assessed by quantitative real-time reverse transcription PCR. The expression patterns of OsZIP1, OsZIP4, and OsZIP5 in root and shoot of rice were altered due to the Zn availability as dictated by Zn sources and ZSB inoculation. Fe sufficiency significantly reduced the root and shoot OsZIP1 expression, but not the OsZIP4 and OsZIP5 levels. Zinc oxide in the growth medium up-regulated all the assessed ZIP genes in root and shoot of rice seedlings. When ZSB was inoculated to rice seedlings grown with insoluble zinc oxide in the growth medium, the expression of root and shoot OsZIP1, OsZIP4, and OsZIP5 was reduced. In the absence of zinc oxide, ZSB inoculation up-regulated OsZIP1 and OsZIP5 expressions. Zinc nutrition provided to the rice seedling through ZSB-bound zinc oxide solubilization was comparable to the soluble zinc sulfate application which was evident through the ZIP genes’ expression and the Zn accumulation in root and shoot of rice seedlings. These results demonstrate that ZSB could play a crucial role in zinc fertilization and fortification of rice.

Desiccation survival of Acinetobacter spp. in infant formula

Acinetobacter spp. are included under category B in the FAO-WHO list of organisms of concern for neonatal health following the consumption of powdered infant formula. However, the ability of Acinetobacter spp. to maintain their viability in desiccated infant formula over a storage period consistent with the shelf-life of commercially available powdered infant formula (2 years) has not been demonstrated. In this study, 9 clinical and food isolates of Acinetobacter baumannii, A. calcoaceticus, and Acinetobacter genomosp. 3 were desiccated in infant formula and then reconstituted at designated time points. Bacterial viability was followed for a maximum period of 24 months or until the strain became undetectable (<5 × 102 cfu/ml). For comparative purposes, one Enterobacter hormaechei and two Enterobacter cloacae strains were also monitored for their desiccation survival. The seven clinical and food strains remained cultivable for the whole duration of the study and showed biphasic survival curves. The initial drop in viable count was up to 3.5 log10 cfu/ml within 18 h of desiccation exposure. By the end of the study, the reduction in viability was between 3.6 and 4.8 log10 cfu/ml. In contrast the A. baumanii and A. calcoaceticus species type strains only persisted for 6 and 9 months, respectively, possibly due to laboratory adaptation. The E. cloacae and E. hormaechei strains were undetectable after 12 and 17 months, respectively. The persistence of Acinetobacter spp. strains in desiccated powdered infant formula, supports the FAO-WHO designation of this organism as a risk to neonatal health.

Phytases from Enterobacter and Serratia species with desirable characteristics for food and feed applications.

Phytases are enzymes of great industrial importance with wide range of applications in animal and human nutrition. These catalyze the hydrolysis of phosphomonoester bonds in phytate, thereby releasing lower forms of myo-inositol phosphates and inorganic phosphate. Addition of phytase to plant-based foods can improve its nutritional value and increase mineral bioavailability by decreasing nutritional effect of phytate. In the present investigation, 43 phytase positive bacteria on PSM plates were isolated from different sources and characterized for phytase activity. On the basis of phytase activity and zone of hydrolysis, two bacterial isolates (PSB-15 and PSB-45) were selected for further characterization studies, i.e., pH and temperature optima and stability, kinetic properties and effect of modulators. The phytases from both isolates were optimally active at the pH value from 3 to 8 and in the temperature range of 50–70 °C. Further, the stability of isolates was good in the pH range of 3.0–8.0. Much variation was observed in temperature and storage stability, responses of phytases to metal ions and modulators. The K m and V max values for PSB-15 phytase were 0.48 mM and 0.157 μM/min, while for PSB-45 these were 1.25 mM and 0.140 μM/min, respectively. Based on 16S rDNA gene sequence, the isolates were identified as Serratia sp. PSB-15 (GenBank Accession No. KR133277) and Enterobacter cloacae strain PSB-45 (GenBank Accession No. KR133282). The novel phytases from these isolates have multiple characteristics of high thermostability and good phytase activity at desirable range of pH and temperature for their efficient use in food and feed to facilitate hydrolysis of phytate-metal ion complex and in turn, increased bioavailability of important metal ions to monogastric animals.

Zinc Solubilizing Potential of Enterobacter cloacae Strain ZSB14 in Three Different Semi-arid Tropical Soils

Microbial transformation of soil unavailable zinc (Zn) into available form is an important agronomical approach not only to alleviate the Zn deficiency of stable food crops but also to improve the nutritional quality of human diet. Zinc solubilizing bacteria (ZSB) as an inoculant to the crop plants could be a sustainable input for Zn-fertilization as well as for Zn-biofortification. In order to explore these bacteria, we have conducted a microcosm study to assess its potential in terms of Zn availability in three different soils of semi-arid tropics and its interaction effects with different insoluble Zn amendments.....

Potential Bacterial Zoonotic Pathogens Isolated from a Major Abattoir and its Receiving Surface Water in Abeokuta, Nigeria

The rapid pace and scale of urbanization in Nigeria has led to large volumes of domestic and industrial wastes, which pose threats to the environment and public health. Currently, in Nigeria, there seems to be neither sufficient measures nor facilities to treat abattoir effluents for environmental safety or to recover usable energy and material from abattoir by-products. This study was conducted to assess the microbial status of the Lafenwa abattoir effluent and its receiving surface water. The Total viable and Coliform counts (TBC and TCC) using surface plating and multiple tube test techniques, respectively were performed. Bacterial contaminants were isolated and identified by standard microbiological procedures. Furthermore, various methods for waste disposal at the abattoir were assessed. The mean TBC and TCC for waste water during and after slaughtering were 5.2x107, 4.9x107 and 4.26x107, 3.06x107 cfu/ml respectively. The receiving surface water had mean TBC and TCC of 4.15x107, 3.83x107, and 3.89x107, 2.87x107 cfu/ml respectively. Waste disposal in the abattoir was by open dumping of solid wastes while effluent was discharged into the nearby Ogun River, which is also used by butchers for meat processing. Bacterial organisms isolated from the effluent include Enterobacter aerogenes, Hafnia alvei, Erwinia mallotivora, Edwardsiella ictaluri, Enterobacter amnigenus and Escherichia coli O157strains while Proteus miriabilis, Staphylococcus spp, Pseudomonas aeruginosa, Enterobacter intermedius, Yersinia aleksiciae, Serratia odorifera, Enterobacter cloacae, Enterobacter aerogens and Eschericia coli O157 strains were isolated from the surface water. The high microbial load and the isolation of pathogenic organisms of public health importance especially Escherichia coli O157 strains further demonstrates the need for adequate waste disposal and treatment. Also, the current methods of waste disposal at the abattoir greatly reduces the quality of the surrounding environment and portends a risk to public health and food safety.

Inhibition of Penicillium digitatum and Citrus Green Mold by Volatile Compounds Produced by Enterobacter cloacae

Penicillium digitatum causes green mold decay on citrus fruit, resulting in severe economic losses to citrus growers and packers worldwide. The present study is to evaluate the control of citrus green mold by volatiles produced by Enterobacter cloacae. An E. cloacae strain isolated from plant rhizospheres was able to produce three volatile organic compounds, which were identified as butyl acetate, phenylethyl alcohol, and 4,5-dimethyl-1-hexene by GC/MS chromatography. The volatile compounds produced by E. cloacae inhibited conidial germination and hyphal elongation of P. digitatum and reduced green mold severity. E. cloacae cultured at temperatures ranging from 16°C to 28°C, at pH values ≤6, or in a substrate carrier (sphagnum moss, vermiculite, or perlite) provided superior control against P. digitatum. A laboratory formulation using E. cloacae and perlite protected citrus fruit from green mold up to 22 days and its effectiveness outperformed fungicide application at room temperature (~25°C). The results implicate practical application of E. cloacae as a biofumigant for controlling citrus postharvest decay caused by P. digitatum. Significantly, the study provides a model for future research on how to formulate an effective biocontrol agent for disease management.

CTX-M-15형 Extended Spectrum β-lactamase와 ArmA 동시 생성 Enterobacter cloacae의 출현

We investigated the prevalence of extended spectrum -lactamase (ESBL) genes and 16S rRNA methyltransferase genes to study antimicrobial resistance mechanisms of Enterobacter cloacae strains isolated from a university hospital in the Chungcheong province of Korea. Eight of the bacteria strains involved in this study contained CTX-M-15 type ESBL. Among 8 strains harboring the ESBL gene, 3 strains also harbored armA gene. The three isolates showed resistance to antimicrobial agents belonged to third cephalosporin, aminoglycoside, and fluoroquinolones. Furthermore, interspecies plasmid transfer of the antimicrobial resistant genes may induced horizontal spreading of the genes and emergence of multidrug resistant bacteria. Therefore, surveillance for existence of antimicrobial resistance determinants is important to prevent distribution of antimicrobial resistant strains.

Characteristics of a Novel Aerobic Denitrifying Bacterium, Enterobacter cloacae Strain HNR

A novel aerobic denitrifier strain HNR, isolated from activated sludge, was identified as Enterobacter cloacae by16S rRNA sequencing analysis. Glucose was considered as the most favorable C-source for strain HNR. The logistic equation well described the bacterial growth, yielding a maximum growth rate (μmax) of 0.283h(-1) with an initial NO3 (-)-N concentration of 110mg/L. Almost all NO3 (-)-N was removed aerobically within 30h with an average removal rate of 4.58mgNL(-1)h(-1). Nitrogen balance analysis revealed that proximately 70.8% of NO3 (-)-N was removed as gas products and only 20.7% was transformed into biomass. GC-MS result indicates that N2 was the end product of aerobic denitrification. The enzyme activities of nitrate reductase and nitrite reductase, which are related to the process of aerobic denitrification, were 0.0688 and 0.0054 U/mg protein, respectively. Thus, the aerobic denitrification of reducing NO3 (-) to N2 by strain HNR was demonstrated. The optimal conditions for nitrate removal were C/N ratio 13, pH value 8, shaking speed 127rpm and temperature 30°C. These findings show that E. cloacae strain HNR has a potential application on wastewater treatment to achieve nitrate removal under aerobic conditions.

Spread of Enterobacter cloacae carrying blaNDM-1, blaCTX-M-15, blaSHV-12 and plasmid-mediated quinolone resistance genes in a surgical intensive care unit in Croatia.

The objective of this study was to describe a hospital cluster of NDM-1-producing Enterobacter cloacae infections observed in the surgical intensive care unit (ICU) of a tertiary-care hospital at Pula, Croatia. NDM-1-producing E. cloacae strains isolated from clinical samples were screened by PCR for the presence of carbapenemases. Genetic relatedness of NDM-1-producing E. cloacae strains was determined by multilocus sequence typing (MLST). During the period October 2013 to April 2014, four patients, with overlapping hospital stay in the surgical ICU, developed severe infections caused by E. cloacae demonstrated to produce carbapenemases. According to MLST, all strains belonged to ST133 and were positive by PCR for the blaNDM-1 carbapenemase gene, for blaCTX-M-15 and blaSHV-12 extended-spectrum β-lactamase (ESBL) genes, and for blaTEM-1 and blaOXA-1 narrow-spectrum β-lactamase genes. They were negative for other carbapenemases genes including blaOXA-48, blaVIM and blaKPC as well as for AmpC and the armA and rmtB aminoglycoside resistance genes. All strains were positive for the HI2 replicon, suggesting that an IncHI2 plasmid is likely the plasmid carrying the blaNDM-1 gene. Infection control measures were implemented after the first case although they were not effective in avoiding spread of this organism to other patients in the surgical ICU. In conclusion, the evolving epidemiology of NDM-producing micro-organisms and the interspecies diffusion of this resistance mechanism to emerging pathogens such as E. cloacae necessitate the setting up of strong and urgent joint measures to control the spread of NDM carbapenemase especially in the ICU setting.

Particular Distribution of Enterobacter cloacae Strains Isolated from Urinary Tract Infection within Clonal Complexes.

Background:Based on biochemical properties, Enterobacter cloacae represents a large complex of at least 13 variant species, subspecies, and genotypes that progressively identified as the most species causing hospital-acquired infections. The aim of this study was to determine the relevance between phylogenetically related strains within the E. cloacae complex and the frequency of urinary tract infection caused by them. Methods:A 268-bp fragment was obtained from hsp60 gene for 50 clinical E. cloacae isolates from urine cultures of inpatients that admitted to six hospitals in Tehran, Iran during December 2012 to November 2013. The 107 nucleotide sequences were analyzed and the evolutionary distances of sequences were computed and neighbor-joining tree was calculated. Results:It showed that all of the genetic clusters have not an equal involvement in pathogenesis of urinary tract infections. Three superior clusters were found, together representing more than two third (80%) of the isolates (cluster VI with 25 members; clusters III and VIII with 9 and 6 members, respectively) and some genetic clusters were absent (IV, X, XII, and xiii), some of which are supposed to be associated with plants and no human infection has been reported. Conclusions:This study, for the first time, reports the unequal contribution of E. cloacae complex subspecies and clusters in urinary tract infections in Iran and together with studies from other countries suggest that the subspecies of E.hormaechei subsp. Oharae is the most prevalent E. cloacae complex subspecies regardless of country under study.

Bloodstream Infections due to Enterobacteriaceae Among Neonates in Poland – Molecular Analysis of the Isolates

Bloodstream infections (BSIs) are associated with a significantly increased risk of fatality. No report has been found about the molecular epidemiology of Enterobacteriaceae causing BSI in neonates in Poland. The aim of this work was to determine the antibiotic resistance profiles, virulence gene prevalence, the epidemiological and genetic relationships among the isolates from Enterobacteriaceae causing BSI in neonates with birth weight < 1501 g. Antimicrobial susceptibility testing was performed. PCR was performed to identify the presence of common beta-lactamase genes, virulence genes. PFGE and MLST were performed. The surveillance group contained 1,695 newborns. The incidence rate for BSIs was 5.9%, the fatality rate 15%. The most common species were Escherichia coli (n = 24) and Klebsiella pneumoniae (n = 16). CTX-M-15 was found in 6 E. coli, 8 K. pneumoniae, 1 Enterobacter cloacae strains. Among E. coli fimH (83.3%), ibeA (37.5%), neuC (20.8%) were the most frequent. PFGE demonstrated unique pulsotypes among E. coli. E. coli ST131 clone was found in 7 E. coli strains. PFGE of 16 K. pneumoniae strains showed 8 pulsotypes. Five isolates from one NICU belonged to one clone. MLST typing revealed 7 different ST with ST336 as the most prevalent. This study provides information about resistance, virulence and typing of Enterobacteriaceae strains causing BSI among neonates. E. coli and Klebsiella spp. isolated in this study have completely different epidemiology from each other.

Characterization of an Enterobacter cloacae Strain Producing both KPC and NDM Carbapenemases by Whole-Genome Sequencing.

A carbapenem-resistant Enterobacter cloacae strain, WCHECl-14653, causing a fatal bloodstream infection, was characterized by genome sequencing and conjugation experiments. The strain carried two carbapenemase genes, blaNDM-1 and blaKPC-2, on separate IncF plasmids. The coexistence of blaNDM-1 and blaKPC-2 conferred slightly higher-level carbapenem resistance compared with that of blaNDM-1 or blaKPC-2 alone, and the coexistence of two IncF plasmids may generate new platforms for spreading carbapenemase genes.

Identification of sdiA-regulated genes in a mouse commensal strain of Enterobacter cloacae.

Many bacteria determine their population density using quorum sensing. The most intensively studied mechanism of quorum sensing utilizes proteins of the LuxI family to synthesize a signaling molecule of the acylhomoserine lactone (AHL) type, and a protein of the LuxR family to bind AHL and regulate transcription. Genes regulated by quorum sensing often encode functions that are most effective when a group of bacteria are working cooperatively (e.g., luminescence, biofilm formation, host interactions). Bacteria in the Escherichia, Salmonella, Klebsiella, and Enterobacter genera do not encode an AHL synthase but they do encode an AHL receptor of the LuxR family, SdiA. Instead of detecting their own AHL synthesis, these organisms use SdiA to detect the AHLs synthesized by other bacterial species. In this study, we used a genetic screen to identify AHL-responsive genes in a commensal Enterobacter cloacae strain that was isolated from a laboratory mouse. The genes include a putative type VI secretion system, copA (a copper transporter), and fepE (extends O-antigen chain length). A new transposon mutagenesis strategy and suicide vectors were used to construct an sdiA mutant of E. cloacae. The AHL-responsiveness of all fusions was entirely sdiA-dependent, although some genes were regulated by sdiA in the absence of AHL.

Prevalence of fecal carriage of extended-spectrum- and metallo-β-lactamase-producing gram-negative bacteria among neonates born in a hospital setting in central Saudi Arabia.

BACKGROUND AND OBJECTIVECommensal neonatal fecal flora constitute a reservoir of antibiotic resistance. The aim of this study was to characterize the prevalence of fecal carriage of extended spectrum beta-lactamases (ESBLs) and carbapenemase producing gram-negative bacteria among 150 neonates who were born in two hospitals in central Saudi Arabia.PATIENTS AND METHODSFrom June 2012 to Janauary 2013, 150 healthy neonates ≤7 days of age were screened for intestinal enteric bacteria colonization and the influence of age, mode of delivery, body weight and breast feeding of neonates on colonzation rates. The isolates were identified manually and by VITEK 2 and antibiotic susceptibility was tested. Phenotypic tests were performed to seek ESBL and metallo-β-lactamase (MBL) producers in recovered isolates.RESULTSAmong 150 fecal specimens, 188 gram-negative rods were recovered. E coli (n=130; 69.1%) was the commonest organism followed by Klebsiella pneumoniae (n=23; 12.2%), Pseudomonas aeuroginosa (n=14; 7.5%), Acinetobacter baumannii (n=9; 4.8%), Enterobacter spp. (n=9; 4.8%) and Morganella morganii (n=3; 1.6%). E coli colonization during the first day of life was 95.5% (n=43/45), which was relatively higher than in subsequent days. The colonization rate of E coli among low weight babies (≤3 kg) was significantly higher than fully grown neonates (≥4 kg)(P=.03). The prevalence of ESBL and MBL was 15.0% (n=28/188) and 1.6% (n=3/188). The prevalence rates of ESBL in E coli, K pneumoniae, E cloacae, M morganii and E aerogenes were 16.9% (n=22/130), 4.3 % (n=1/23), 60% (n=3/5), 33.3% (n=1/3), and 25% (n=1/4), respectively. All E cloacae with ESBLs and MBLs were resistant to all tested antibiotics including carbapenems.CONCLUSIONWithin the first week of life, the most important determinants of gut enteric bacteria composition in neonates were age in days, body weight, mode of delivery, and type of neonatal feeding. The findings of ESBLs- and MBLs-producing enteric bacteria as neonate-gut colonizers may have wider implication or linkage to prevention of nosocomial and community-acquired infections by these strains.

Draft Genome Sequence of Enterobacter cloacae Strain S611.

We report draft genomes of Enterobacter cloacae strain S611, an endophytic bacterium isolated from surface-sterilized germinating wheat seeds. We present the assembly and annotation of its genome, which may provide insights into the metabolic pathways involved in adaptation.

Metabolic engineering of Enterobacter cloacae for high-yield production of enantiopure (2R,3R)-2,3-butanediol from lignocellulose-derived sugars

Biotechnological production of biofuels is restricted by toxicity of the products such as ethanol and butanol. As its low toxicity to microbes, 2,3-butanediol (2,3-BD), a fuel and platform bio-chemical, could be a promising alternative for biofuel production from renewable bioresources. In addition, no bacterial strains have been reported to produce enantiopure 2,3-BD using lignocellulosic hydrolysates. In this study, Enterobacter cloacae strain SDM was systematically and metabolically engineered to construct an efficient biocatalyst for production of the fuel and enantiopure bio-chemical—(2R,3R)-2,3-BD. First, the various (2R,3R)-2,3-BD dehydrogenase encoding genes were expressed in a meso-2,3-BD dehydrogenase encoding gene disrupted E. cloacae strain under native promoter Pb of the 2,3-BD biosynthetic gene cluster of E. cloacae. Then, carbon catabolite repression was eliminated via inactivation of the glucose transporter encoding gene ptsG and overexpression of a galactose permease encoding gene galP. The resultant strain could utilize glucose and xylose simultaneously. To improve the efficiency of (2R,3R)-2,3-BD production, the byproduct-producing genes (ldh and frdA) were knocked out, thereby enhancing the yield of (2R,3R)-2,3-BD by 16.5% in 500-mL Erlenmeyer flasks. By using fed-batch fermentation in a 5-L bioreactor, 152.0g/L (2R,3R)-2,3-BD (purity>97.5%) was produced within 44h with a specific productivity of 3.5g/[Lh] and a yield of 97.7% from a mixture of glucose and xylose, two major carbohydrate components in lignocellulosic hydrolysates. In addition, when a lignocellulosic hydrolysate was used as the substrate, 119.4g/L (2R,3R)-2,3-BD (purity>96.0%) was produced within 51h with a productivity of 2.3g/[Lh] and a yield of 95.0%. These results show that the highest records have been acquired for enantiopure (2R,3R)-2,3-BD production by a native or engineered strain from biomass-derived sugars. In addition to producing the 2,3-BD, our systematic approach might also be used in the production of other important chemicals by using lignocellulose-derived sugars.

Diversity of carbapenemases in clinical isolates of Enterobacteriaceae in Croatia—the results of a multicentre study

Since the first carbapenem-resistant Klebsiella pneumoniae strain was isolated in 2008, Enterobacteriaceae with reduced susceptibility to one or more carbapenems have emerged sporadically in different geographical regions in Croatia. These observations gave rise to a multicenter study on carbapenem resistance in Enterobacteriaceae from Croatia. Fifty-seven carbapenem-non-susceptible strains of Enterobacteriaceae were collected during 2011–2012 from four large hospital centres in Croatia. Overall, 36 strains produced VIM-1 β-lactamase, three produced NDM-1, and one produced KPC-2. A high degree of clonal relatedness was observed in Enterobacter cloacae and Citrobacter freundii strains, in contrast to K. pneumoniae strains. BlaVIM genes were located within class1 integron which contained genes encoding resistance to aminoglycosides (aacA4). The study found strong association between blaVIM and qnrB6 and between blaNDM and qnrA6 genes.

Isolation and identification of an Enterobacter cloacae strain producing a novel subtype of Shiga toxin type 1.

We describe here the isolation and identification of a Shiga toxin 1 (Stx1)-producing Enterobacter cloacae strain, M12X01451, from a human clinical specimen. The bacterial isolate was identified as E. cloacae using a polyphasic approach that included phenotypic, genetic, and proteomic analyses. The M12X01451 stx1 was sequenced, and the holotoxin was found to share only 87% amino acid sequence identity with the nearest Stx1 subtype reference sequence. Sequence analysis of the regions immediately flanking stx1 displayed similarities with bacteriophage-related sequences, suggesting a prophage origin. The stx1 gene was a stable element within the M12X01451 genome, as demonstrated by real-time PCR detection following successive subculturing of the bacterial isolate. Culture supernatant from M12X01451 was cytotoxic to Vero cells but was not neutralized by an anti-Stx1 monoclonal antibody. In addition, Stx1 from M12X01451 demonstrated limited antigenicity with two commercially available lateral flow immunoassays. The M12X01451 Stx represents a new Stx1 subtype based on the degree of sequence dissimilarity with Stx1 subtype reference sequences and its limited reactivity with anti-Stx1 antibodies.

Molecular typing and characterization of TEM, SHV, CTX-M, and CMY-2 β-lactamases in Enterobacter cloacae strains isolated in patients and their hospital environment in the west of Algeria

ObjectivesEnterobacter cloacae is a major nosocomial bacterium causing severe infections. A multicenter retrospective cohort study was conducted to collect baseline information on the molecular characteristics of β-lactamase producing Enterobacter cloacae in the west of Algeria. Materials and methodsWe report a series of 42 extended-spectrum β-lactamase (ESBL) producing non-repetitive Enterobacter cloacae strains, collected in 3 university hospital (Tlemcen, Oran, and Sidi Bel Abbes). Antibiotic susceptibility testing (antibiogram and MIC) and screening for ESBL were performed according to the French Society for Microbiology guidelines. PFGE typing was used to characterize the clonality of all the strains. β-lactamase genes (blaTEM, blaSHV, blaCTX-M, blaECB, and blaCMY-2) were amplified by PCR with specific primers. Plasmid isolation, electroporation, and conjugation experiments were carried out using standard methods. ResultsSequence analysis revealed that most strains produced CTX-M type ESBLs (CTX-M-15 and CTX-M-3), whereas only 5 produced SHV-type ESBLs (SHV-12). The blaTEM gene was identified in all strains of Enterobacter cloacae. Several epidemic clones were determined. One strain was found to produce plasmid-mediated AmpC β-lactamase (CMY-2); this gene was transferred from E. cloacae by electroporation. Conjugation experiments showed that blaCTX-M, blaTEM, and blaSHV were carried by conjugative plasmids of high molecular weight (≥70kb). ConclusionThe emergence of resistance genes is a public health problem.

Complete genome of the switchgrass endophyte Enterobacter clocace P101.

The Enterobacter cloacae complex is genetically very diverse. The increasing number of complete genomic sequences of E. cloacae is helping to determine the exact relationship among members of the complex. E. cloacae P101 is an endophyte of switchgrass (Panicum virgatum) and is closely related to other E. cloacae strains isolated from plants. The P101 genome consists of a 5,369,929 bp chromosome. The chromosome has 5,164 protein-coding regions, 100 tRNA sequences, and 8 rRNA operons.