Cloacae Strain Research Articles

Characterization and genomic analysis of the lytic bacteriophage vB_EclM_HK6 as a potential approach to biocontrol the spread of Enterobacter cloacae contaminating food

BackgroundIncreased prevalence of Enterobacter cloacae within food products underscores food as an underexplored reservoir for antibiotic resistance, thus requiring particular intervention. Bacteriophages have been explored as a promising approach for controlling bacterial growth in different matrices. Moreover, their specific interaction and self-replication, put them apart from traditional methods for controlling bacteria in different matrices.MethodsSixteen Enterobacter cloacae strains were recovered from raw chicken. These strains were used to isolate bacteriophages using enrichment protocol. The broad-spectrum bacteriophage was evaluated in terms of thermal, pH, shearing stress and storge. Moreover, its infection kinetics, in vitro antibacterial activity, cytotoxicity were also assessed. Genomic sequencing was performed to exclude any potential virulence or resistance genes. Finally, the capability of the isolated phages to control bacterial growth in different chicken samples was assessed alone and in combination with sodium nitrite.ResultsThe lytic bacteriophage vB_EclM_HK6 was isolated and showed the broadest spectrum being able to infect 8/16 E. cloacae strains with a lytic activity against its host strain, E. cloacae EC21, as low as MOI of 10–6. The phage displays a latent period of 10 min and burst size of 115 ± 44 and resistance frequency of 5.7 × 10–4 ± 3.0 × 10–4. Stability assessment revealed a thermal tolerance up to 60 ˚C, wide range pH stability (3–10) and the ability to withstand shearing stress up to 250 rpm. HK6 shows no cytotoxicity against oral epithelial cells up to 1012 PFU/ml. Genomic analysis revealed a Strabovirus with total size of 177,845 bp that is free from known resistance and virulence genes. Finally, HK6 pretreatment of raw chicken, chicken nuggets and ready-made cheese salad shows a reduced bacterial count up to 4.6, 2.96 and 2.81 log-units, respectively. Moreover, combing HK6 with sodium nitrite further improved the antibacterial activity in both raw chicken and chicken nuggets without significant enhancement in case of cheese salad.ConclusionEnterobacter bacteriophage vB_EclM_HK6 presents a safe and effective approach for controlling E. cloacae contaminating stored chicken food samples. Moreover, they could be combined with a reduced concentrations of sodium nitrite to improve the killing capacity.

Evaluation of the antibacterial effect of (R)-(+)-Limonene against Enterococcus faecalis and Enterobacter cloacae strains isolated from food

The aim of this study was to evaluate the potential antibacterial and anti-adherent activities of the monoterpene (R)-(+)-limonene, as well as its synergistic potential with synthetic antimicrobials against strains of Enterococcus faecalis and Enterobacter cloacae. The antibacterial properties of (R)-(+)-limonene were assessed using the broth microdilution technique to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Additionally, the infusion disc method was employed to explore the association of the compound with antimicrobials, and the test tube method was used to determine the minimum inhibitory concentration of adherence (MICA). It was observed that the MIC for (R)-(+)-limonene was 1000 μg mL-1 for five of the six E. faecalis strains tested, while for E. cloacae, the MIC exceeded 1000 μg mL-1 for all strains tested. Identical values were recorded for the MBC in E. faecalis. In terms of its combination with synthetic antimicrobials, (R)-(+)-limonene demonstrated a synergistic effect with gentamicin and ciprofloxacin for most strains. Regarding the MICA, both (R)-(+)-limonene and 0.12% chlorhexidine digluconate failed to inhibit biofilm formation at the tested concentrations. Given the need for new therapeutic alternatives for treating bacterial infections, this study revealed that the tested monoterpene exhibited moderate bactericidal effects against E. faecalis strains and no antibacterial effect against E. cloacae strains. However, when combined with various classes of antimicrobials, (R)-(+)-limonene showed synergistic effects with gentamicin and ciprofloxacin for most strains. This suggests that (R)-(+)-limonene holds promise for enhancing the treatment of bacterial infections and could support conventional therapies. Nonetheless, further in vitro, ex vivo, and in vivo studies are necessary to confirm and elucidate its efficacy and mechanisms.

Influences of Enterobacter Cloacae Strain Fg 5-2 and Its Vermicompost and Nitrogen Fertilizer Usage Efficiency on Groundnut Yield

Influences of Enterobacter Cloacae Strain Fg 5-2 and Its Vermicompost and Nitrogen Fertilizer Usage Efficiency on Groundnut Yield

Mining a bioremediation genetic toolbox in the novel environmental bacteria Enterobacter cloacae strain amazonensis

Mining a bioremediation genetic toolbox in the novel environmental bacteria Enterobacter cloacae strain amazonensis

Biological Characterization and Genomic Analysis of Three Novel Serratia- and Enterobacter-Specific Virulent Phages.

Due to the high microbiological contamination of raw food materials and the increase in the incidence of multidrug-resistant bacteria, new methods of ensuring microbiological food safety are being sought. One solution may be to use bacteriophages (so-called phages) as natural bacterial enemies. Therefore, the aim of this study was the biological and genomic characterization of three newly isolated Serratia- and Enterobacter-specific virulent bacteriophages as potential candidates for food biocontrol. Serratia phage KKP_3708 (vB_Sli-IAFB_3708), Serratia phage KKP_3709 (vB_Sma-IAFB_3709), and Enterobacter phage KKP_3711 (vB_Ecl-IAFB_3711) were isolated from municipal sewage against Serratia liquefaciens strain KKP 3654, Serratia marcescens strain KKP 3687, and Enterobacter cloacae strain KKP 3684, respectively. The effect of phage addition at different multiplicity of infection (MOI) rates on the growth kinetics of the bacterial hosts was determined using a Bioscreen C Pro growth analyzer. The phages retained high activity in a wide temperature range (from -20 °C to 60 °C) and active acidity values (pH from 3 to 12). Based on transmission electron microscopy (TEM) imaging and whole-genome sequencing (WGS), the isolated bacteriophages belong to the tailed bacteriophages from the Caudoviricetes class. Genomic analysis revealed that the phages have linear double-stranded DNA of size 40,461 bp (Serratia phage KKP_3708), 67,890 bp (Serratia phage KKP_3709), and 113,711 bp (Enterobacter phage KKP_3711). No virulence, toxins, or antibiotic resistance genes were detected in the phage genomes. The lack of lysogenic markers indicates that all three bacteriophages may be potential candidates for food biocontrol.

Removal of residual organic pollutants from pulp paper industry wastewater using bacterial strains Acinetobacter baumannii (OK582199) PC4 and Bacillus cereus (OK582201) PC10 via biostimulation and bioaugmentation process

This study examines how native bacteria, combined with two specific aerobic strains (Acinetobacter baumannii PC4 and Bacillus cereus PC10), contribute to the detoxification of pulp and paper industrial wastewater (PPIW) through biostimulation and bioaugmentation processes. The growth of two native bacteria were stimulated in pulp paper industry wastewater by the addition of carbon (1% glucose) and nitrogen (0.5% peptone w/v). After the growth of these native bacteria in biostimulation process showed significant reduction in various physico-chemical parameters i.e. lignin (53%), color (46%), COD (76%), and BOD (86%). Similarly, reductions in various heavy metals were also observed, such as Fe (73%), Zn (36%), Cr (41%), Cu (22%), and Ni (65%). The growing native bacterial co-culture were identified as Enterobacter cloacae strain PC7 (OQ672613) and Bacillus sp. strain PCB (OK614116). However, the addition of Acinetobacter baumannii (OK582199) PC4 and Bacillus cereus (OK582201) PC10 enhanced the reduction of various physico-chemical parameters in range of 10–––28%. Consequently, the UV–Vis absorption peak showed reduction due to the decolorization of wastewater and degradation of various pollutants. Moreover, GC–MS analysis indicated that most of the compounds detected in the control were diminished after bacterial treatment. However, compounds such as Pentadecane, 2-Methyl, Silane (dodecylosy) trimethyl, Octadecanoic acid, trimethylsilyl ester, and Docasane were noted as recalcitrant. Additionally, new metabolic products such as 1-ethyl-5,6-dimethoxy-2,3-dihydroindole, Trimethylsily ether of glycerol, and 1-(Trimethylsilylmethyl1) dimethylsilyloxy tetradecane were detected. Due to degradation of various pollutants toxicity reductions were found up to 76% during a comparative evaluation between control and bacterial treated wastewater using the Phaseolus mungo L. seed germination test. Hence, it was concluded that the biostimulation and bioaugmentation process would be an effective technique for degrading residual organic pollutants from pulp and paper industrial wastewater (PPIW) for safe disposal.

The Abundance of Plasmid-Mediated Quinolone Resistance Genes in Enterobacter cloacae Strains Isolated from Clinical Specimens in Kermanshah, Iran.

Enterobacter cloacae (E. cloacae) is one of the most common Enterobacteriaceae causing nosocomial infections. Plasmid-mediated quinolone resistance (PMQR) determinants have been considered recently. This study evaluated the abundance of PMQR genes in strains of E. cloacae obtained from clinical samples in Kermanshah, Iran. In this descriptive cross-sectional study, after collecting 113 isolates of E. cloacae, their identity was confirmed using specific biochemical tests. After determining their drug resistance patterns using disc diffusion, the phenotypic frequency of extended-spectrum beta-lactamase (ESBL)-producing isolates was measured by the double-disk synergy test (DDST) method. The isolates were examined for the presence of qnrA, qnrB, qnrS, and aac(6')-Ib-cr genes by the polymerase chain reaction (PCR) assay. The antibiotic resistance rate of E. cloacae isolates varied from 9.7% to 60.2%; among them, 78% were multidrug-resistant (MDR). The highest quinolone resistance was observed in ESBL-producing strains of E. cloacae. The frequency of positive isolates for PMQR and ESBL was 79.6% and 57.5%, respectively. The genes aac(6')-ib-cr (70.8%) and qnrB (38.1%) had the highest frequency among other genes. The number of isolates simultaneously carrying 2 and 3 genes was 64 and 5 isolates, respectively. The obtained results indicate a high degree of quinolone resistance among ESBL-producing E. cloacae strains. Nevertheless, there was a significant relationship between the PMQR gene and ESBL-positive isolates. Therefore, special attention should be paid to molecular epidemiological studies on antibiotic resistance to quinolones and beta-lactamases in these strains.

Enhancing iron bioavailability in lentil grains circumventing the phytate barrier

The study aimed at examining the effect of Iron (Fe) management strategies on the redistribution of Fe fractions in soil and on the Fe bioavailability and phytic acid (PA) content in lentil grains. A pot experiment was conducted in Completely Randomized Design with 8 treatments replicated thrice. The treatments were Control (no fertilizer); Recommended dose of fertilizers (RDF) i.e. 20:40 (N: P2O5); RDF + Fe (basal application @10 kg/ha); RDF + Fe (foliar@ 1% (w/v) solution); RDF + Fe (basal + foliar); RDF + FeSB (biopriming with Fe solubilizing bacteria (Enterobacter cloacae strain BAU3) @ 5 mL/kg seed (1 x109 cfu mL−1)); RDF + Fe (seed nutripriming with 1% (w/v) solution); and RDF + (biopriming + nutripriming). Fe source for the treatments was Fe-EDDHA chelate. Higher Fe release from these fractions was observed under biopriming + nutripriming with water soluble Fe of 21.09 and 20.82 mg kg−1, respectively for the biopriming + nutripriming and sole biopriming treatments; and organically bound Fe of 230.95 and 211.36 mg kg−1 for the same treatments. These fractions contributed toward higher Fe uptake which was highest under biopriming + nutripriming. Values were statistically significant for biopriming and nutripriming and basal + foliar treatment. Lowest PA values in lentil grains as well as PA/Fe ratio were estimated under basal + foliar and biopriming + nutripriming treatments. Combined basal and foliar application of Fe esp. in chelated form and use of seed priming strategies have immense potential in enhancing the Fe bioavailability in grains from soil, seed or foliage.

In vitro activity of human defensins HNP-1 and hBD-3 against multidrug-resistant ESKAPE Gram-negatives of clinical origin and selected peptidoglycan recycling-defective mutants.

In the current scenario of critical need for new antimicrobials against multidrug-resistant bacteria, all options must be considered, including classic ideas such as the use of purified immune compounds. However, information regarding the activity of certain human defensins against ESKAPE Gram-negatives was incomplete. This is the first study comparatively assessing the in vitro activity of two membrane-permeabilizing/peptidoglycan construction-blocking defensins (HNP-1 and hBD-3) against relevant clinical collections of ESKAPE Gram-negatives, alone or in combination with permeabilizers, additional peptidoglycan-targeting attacks, or the blockade of its recycling. Our data suggest that hBD-3 has a notable bactericidal activity against multidrug-resistant Acinetobacter baumannii and Enterobacter cloacae strains that should be considered as potential adjuvant option. Our results suggest for the first time an increased resistance of Pseudomonas aeruginosa strains from chronic infection compared to acute origin ones, and provide new clues about the predominant mode of action of hBD-3 against Gram-negatives (permeabilization rather than peptidoglycan-targeting).

Mechanisms involved in plant growth promotion by Enterobacter cloacae DJ under salinity-alkalinity stress

BackgroundPlant-promoting bacteria are safer alternatives to pesticides and fertilizers, reduce environmental pollution, and increase crop yields. We isolated an Enterobacter cloacae strain DJ with plant-promoting effects from the rhizosphere soil of a plant (Leymus chinensis (Trin.) Tzvel) in the western region of Jilin Province of China and investigated the mechanisms underlying the adaptation of the DJ bacteria to salinity-alkalinity environments and the molecular mechanisms of the cross-talk between DJ bacteria and cucumber seedlings.ResultsThe average diameter of the colonies on the salinity-alkalinity medium after incubation for 24 h was 3.3 cm, and this was significantly higher than the 1.9 cm diameter in ADF medium (p < 0.01). Comparative proteomic analysis revealed that 188 differentially expressed proteins, comprising 116 upregulated and 72 downregulated proteins, significantly changed in salinity-alkalinity groups compared to the control groups. The top one upregulated pathway of KEGG enrichment was bacterial chemotaxis, DJ bacteria adapted to salinity-alkalinity environments by upregulating the genes associated with bacterial chemotaxis. The contents of putrescine in salinity-alkalinity and control groups were 4.73 μg/mL and 3.02 μg/mL, whereas the contents of spermidine were 46.18 ng/mL and 0.34 ng/mL, respectively. Comparing to the control cohorts, the concentrations of both polyamines in the experimental cohorts exhibited statistically significant increases (p < 0.01). The expression of Pt gene encoding polyamine transporter protein was sharply up-regulated in cucumber roots after treatment with DJ bacteria under salinity-alkalinity stress; the expression was more than tenfold higher than that in the control groups. The enzyme activities of POD, SOD, and CAT in cucumber seedlings were higher compared to those in the control groups (p < 0.01). The stem height, number of lateral roots, and fresh weight of cucumber seedlings in the DJ treatment groups were 6.0 cm, 17 roots, and 0.42 g, respectively, whereas those of the control groups were 3.8 cm, 14 roots, and 0.28 g, respectively, with a notable difference between two cohorts (p < 0.01).ConclusionsDJ bacteria can live in salinity-alkalinity conditions by upregulating the expression of genes associated with chemotaxis. The resistance of cucumber seedlings under salinity-alkalinity conditions through the antioxidant pathway was increased by polyamines produced by DJ bacteria.Graphical

Motility behavior and physiological response mechanisms of aerobic denitrifier, Enterobacter cloacae strain HNR under high salt stress: Insights from individual cells to populations

The motility behaviors at the individual-cell level and the collective physiological responsive behaviors of aerobic denitrifier, Enterobacter cloacae strain HNR under high salt stress were investigated. The results revealed that as salinity increased, electron transport activity and adenosine triphosphate content decreased from 15.75 μg O2/g/min and 593.51 mM/L to 3.27 μg O2/g/min and 5.34 mM/L, respectively, at 40 g/L, leading to a reduction in the rotation velocity and vibration amplitude of strain HNR. High salinity stress (40 g/L) down-regulated genes involved in ABC transporters (amino acids, sugars, metal ions, and inorganic ions) and activated the biofilm-related motility regulation mechanism in strain HNR, resulting in a further decrease in flagellar motility capacity and an increase in extracellular polymeric substances secretion (4.08 mg/g cell of PS and 40.03 mg/g cell of PN at 40 g/L). These responses facilitated biofilm formation and proved effective in countering elevated salt stress in strain HNR. Moreover, the genetic diversity associated with biofilm-related motility regulation in strain HNR enhanced the adaptability and stability of the strain HNR populations to salinity stress. This study enables a deeper understanding of the response mechanism of aerobic denitrifiers to high salt stress.

Therapeutic Potential of a Novel Lytic Phage, vB_EclM_ECLFM1, against Carbapenem-Resistant Enterobacter cloacae.

The global rise of multidrug-resistant Enterobacter cloacae strains, especially those that are resistant to carbapenems and produce metallo-β-lactamases, poses a critical challenge in clinical settings owing to limited treatment options. While bacteriophages show promise in treating these infections, their use is hindered by scarce resources and insufficient genomic data. In this study, we isolated ECLFM1, a novel E. cloacae phage, from sewage water using a carbapenem-resistant clinical strain as the host. ECLFM1 exhibited rapid adsorption and a 15-min latent period, with a burst size of approximately 75 PFU/infected cell. Its genome, spanning 172,036 bp, was characterized and identified as a member of Karamvirus. In therapeutic applications, owing to a high multiplicity of infection, ECLFM1 showed increased survival in zebrafish infected with E. cloacae. This study highlights ECLFM1's potential as a candidate for controlling clinical E. cloacae infections, which would help address challenges in treating multidrug-resistant strains and contribute to the development of alternative treatments.

In vitro evaluation of tigecycline synergy testing with nine antimicrobial agents against Enterobacter cloacae clinical strains.

Enterobacterales (especially carbapenem-resistant) are considered an urgent threat to public health. The available antibiotic therapy is limited due to the increase of multidrug-resistant (MDR) strains. Tigecycline, a minocycline derivative, has emerged as a potential key agent in the treatment of MDR isolates. The aim of the study was to evaluate the synergistic effect of tigecycline in combination with nine antimicrobial agents-ceftazidime/avibactam, colistin, ertapenem, gentamicin, imipenem, levofloxacin, meropenem/vaborbactam, polymyxin B, and rifampicin. Eighty clinical Enterobacter cloacae strains were obtained from patients of two University Hospitals in Bialystok, Poland. The E-test method was used to determine synergistic interactions. Among all combinations, synergy was reported in 61% of cases, addition in 32%, and indifference in 7%. The highest synergy rates were observed in tigecycline combinations with: ceftazidime/avibactam (60/80; 75%), imipenem (60/80; 75%), polymyxin B (55/80; 68.75%) and rifampicin (55/80; 68.75%), while the lowest synergy rate was noted in tigecycline-levofloxacin (26/80; 32.5%). The tigecycline-gentamicin showed the highest rate of indifference; antagonism, was not observed in any combination. In conclusion, tigecycline appears more suitable for use in combination therapy rather than as monotherapy and can be effectively paired with various antimicrobial agents against MDR E. cloacae. Further research will be necessary to confirm these results.

Sustainable approach for the degradation of Contrast dye Evans blue by Enterobacter cloacae strain SD4-1

BackgroundDischarge of dye complexes through industrial effluents continues to be a never-ending problem for the environment. Several degradation techniques have been experimented with, of which utilizing biological entities for the effective degradation of these dye molecules is extensively studied and still being explored. The present study focuses on (i) isolation of bacterial strains from the contaminated agricultural lands (ii) screening the isolates for their decolourization efficiency of Evans blue dye (iii) optimization of culture parameters such as temperature, pH, concentration of dye and nitrogen source for effective decolourization (iv) analysing the effect of degradation of diazo dye Evan's blue by the selected strain using GCMS analysis. MethodsThe bacterial strains were isolated from contaminated agricultural lands in Chennai, Tamil Nadu, India. The collected samples were aseptically transferred to the laboratory and stored for further studies. The collected samples were serially diluted, and the colonies with distinct morphology were pure-cultured. The isolated strains were studied for their efficiency to decolourize the dye Evans blue dye. The strain with higher decolourization efficiency was selected and optimized for parameters such as temperature, pH, nitrogen source and concentration of dye. The degradation of dye by the isolate was analysed by GCMS. Significant FindingsThe investigation unveiled that the strain SD4-1 of Enterobacter cloacae effectively decolorizes Evans blue dye at 37°C and pH 6.0, utilizing Sodium nitrate as the nitrogen source. SD4-1 exhibited an impressive dye decolourization efficiency of approximately 75%. The comprehensive GCMS analysis verified the bacterium's capability to fully degrade the azo dye Evans blue. Consequently, this study concludes that the SD4-1 Enterobacter cloacae strain holds promising potential for the efficient biodegradation of Evan's blue dye. Its application as a highly effective biological agent in the practical removal of dyes from textile industries is envisaged. Notably, this organism demonstrates superior performance in comparison to other studies where a mixture of dyes or consortia of microorganisms were employed for dye decolourization.

Improved application of immobilized Enterobacter cloacae into a bio-based polymer for Reactive Blue 19 removal, an eco-friendly advancement in potential decolorizing systems.

The widespread use of highly complex synthetic dyes like reactive dyes in the textile industry has some adverse environmental impacts and deserves close attention. Biological treatment of these effluents utilizing various species of bacteria with remarkable efficiency in dye removal is still considered promising. Our current study deals with immobilizing an isolated bacterial strain into calcium alginate (Ca/Alg) gel beads and using it to treat pernicious pollutants like synthetic dyes. A potential Reactive Blue 19 (RB19)-degrading Enterobacter cloacae strain A1 was isolated from the Kashan textile industry and was characterized by 16S rDNA gene sequencing. The decolorization ability of strain A1 was assessed by time-based studies using free bacterial cells/immobilized in Ca/Alg. Based on the results of the 16S rDNA gene sequencing, it appears that strain A1 belonged to E.cloacae, with a 99.74% similarity. The findings suggest that immobilized strain A1 accomplished maximum decolorization activity compared with the free cells. The immobilized strain could utterly decompose and decolorize 0.05 mg/mL of RB19 within 48 h, while the free bacterial strain decolorized RB19 within 5 days. Moreover, Ca/Alg gel beads can maintain their efficiency for over three decolorization cycles. Further infrared spectroscopy (FTIR) and gas chromatograph mass spectrometer (GC/MS) investigation declared complete RB19 decomposition on reaction products. Artemia salina was used to investigate the toxicity of dye and its degraded metabolites. The LC50 values signified the pure dye as very toxic with 0.01 mg/mL concentration, while after-treatment products showed no toxic effect on larvae. This immobilization technique increased the applicability of bacterial strain for dye removal. It was beneficial for the decolorization of RB19 from textile wastewater due to a remarkable reduction in time. Notably, strain A1-immobilized beads can maintain their activity for three consecutive decolorization cycles without a considerable decrease in efficiency. PRACTITIONER POINTS: The remarkable capacity of immobilized Enterobacter cloacae strain A1 for Reactive Blue 19 (RB19) removal Immobilized A1 strain showed two-fold higher removal than free one over 48 h A promising method for enhancing RB19 decolorization Decolorization was due to degradation based on UV-Vis, FTIR, and GC/MS analysis Non-toxic posttreatment products for Artemia.

Antibiotic-Resistant Bacteria in Environmental Water Sources from Southern Chile: A Potential Threat to Human Health

Antimicrobial resistance (AMR) is a critical global issue affecting public and animal health. The overuse of antibiotics in human health, animal production, agriculture, and aquaculture has led to the selection of antibiotic-resistant strains, particularly in Gram-negative bacteria. Mutations and horizontal gene transfer play a significant role in the development of antimicrobial resistance, leading to the reduced efficacy of current antibiotics. Today, AMR in bacteria and antibiotic-resistance genes (ARGs) are increasingly recognized in multiple environmental sources, including recreational and irrigation waters. This study aims to identify Gram-negative bacteria from surface aquatic reservoirs in southern Chile and assess their susceptibility to clinically relevant antibiotics. Water samples were collected from four lakes, five rivers, one waterfall, and one watershed in southern Chile to isolate environmental Gram-negative bacilli (GNB). API-20E and MALDI–TOF were employed for bacterial identification. Kirby–Bauer disc diffusion tests and multiplex PCR were performed to determine their susceptibility profile. A total of 26 GNB strains were isolated from environmental water samples, predominantly belonging to the Pseudomonas (n = 9) and Acinetobacter (n = 7) genera. Among these strains, 96.2% were resistant to ampicillin and cefazoline, while 26.9% and 34.6% showed resistance to ceftazidime and cefepime, respectively. Additionally, 38.5% exhibited resistance to colistin. Two Enterobacter cloacae strains obtained from Cachapoal River (sixth region) and Villarrica Lake (ninth region), respectively, presented a multidrug-resistant (MDR) phenotype and carried at least two extended-spectrum β-lactamase (ESBL) genes. Thus, antibiotic-resistant GNB and ARGs were found in natural water reservoirs, raising concerns about the dissemination of resistance determinants among potentially pathogenic bacteria in environmental microbial communities.

Comprehensive Analysis of Virulence Determinants and Genomic Islands of blaNDM-1-Producing Enterobacter hormaechei Clinical Isolates from Greece.

Nosocomial outbreaks of multidrug-resistant (MDR) Enterobacter cloacae complex (ECC) are often reported worldwide, mostly associated with a small number of multilocus-sequence types of E. hormaechei and E. cloacae strains. In Europe, the largest clonal outbreak of blaNDM-1-producing ECC has been recently reported, involving an ST182 E. hormaechei strain in a Greek teaching hospital. In the current study, we aimed to further investigate the genetic make-up of two representative outbreak isolates. Comparative genomics of whole genome sequences (WGS) was performed, including whole genome-based taxonomic analysis and in silico prediction of virulence determinants of the bacterial cell surface, plasmids, antibiotic resistance genes and virulence factors present on genomic islands. The enterobacterial common antigen and the colanic antigen of the cell surface were identified in both isolates, being similar to the gene clusters of the E. hormaechei ATCC 49162 and E. cloacae ATCC 13047 type strains, whereas the two strains possessed different gene clusters encoding lipopolysaccharide O-antigens. Other virulence factors of the bacterial cell surface, such as flagella, fimbriae and pili, were also predicted to be encoded by gene clusters similar to those found in Enterobacter spp. and other Enterobacterales. Secretion systems and toxin-antitoxin systems, which also contribute to pathogenicity, were identified. Both isolates harboured resistance genes to multiple antimicrobial classes, including β-lactams, aminoglycosides, quinolones, chloramphenicol, trimethoprim, sulfonamides and fosfomycin; they carried blaTEM-1, blaOXA-1, blaNDM-1, and one of them also carried blaCTXM-14, blaCTXM-15 and blaLAP-2 plasmidic alleles. Our comprehensive analysis of the WGS assemblies revealed that blaNDM-1-producing outbreak isolates possess components of the bacterial cell surface as well as genomic islands, harbouring resistance genes to several antimicrobial classes and various virulence factors. Differences in the plasmids carrying β-lactamase genes between the two strains have also shown diverse modes of acquisition and an ongoing evolution of these mobile elements.

Bacterial Co-Infection as a Potential Threat to Farmed Flathead Grey Mullet (Mugil cephalus): Phenotypic and Molecular Diagnosis, Histopathology, Immunity Response, and In Vitro Antibacterial Evaluation

Vibrio alginolyticus, Vibrio chlorea, and Enterococcus cloacae were the main bacterial pathogens investigated during recurrent episodes of mortalities among farmed flathead grey mullet (Mugil cephalus) in the Bahar El-Baqar area, Sharqia governorate, Egypt, during the summer season in 2021. Nevertheless, the contamination of water with heavy metals was evaluated. In this study, a total number of sixty moribund and freshly dead Mugil cephalus were examined clinically and upon post-mortem. Then, the bacterial isolates were subjected to bacteriological phenotypic examination, 16S rRNA gene sequencing, and antibiotic sensitivity testing. Histopathological screening and the immunity response of naturally infected M. cephalus were also implemented. In areas where bacterial strains were recovered in a high percentage, water contamination with Cu, Fe, and Ni was higher than the permissible limit. The results showed a high similarity (99–100%) to Vibrio and E. cloacae strains using phylogenetic analysis of 16S rRNA. Most isolates exhibited sensitivity to florfenicol, ciprofloxacin, and norfloxacin, while they were resistant to erythromycin and amoxicillin. Hemorrhage, oedema, hemosiderosis, and melanomacrophage activation in most affected tissues of infected M. cephalus were detected by light microscopy. Nevertheless, higher levels of AST and ALT, as well as serum inflammatory markers, CRP, and lysozyme activity, were shown in infected fish. There were higher MDA levels, along with a reduction of GSH, SOD, and catalase, in the gills and brain of infected M. cephalus, compared to the control fish. Our obtained data confirmed the high prevalence of Vibrio serovars and E. cloacae in infected grey mullets, describing their histopathological alterations, higher inflammatory response, and oxidative stress impact. These results could be useful indicators for understanding the pathogenesis of these pathogens in correlation with water parameter deterioration. Thus, the emergence of Vibrio and E. cloacae species in this study threatens the brackish water aquaculture industry in Egypt and poses a public health concern.

Antimicrobial Resistance Patterns of Enterobacter cloacae and Klebsiella aerogenes Strains Isolated from Clinical Specimens: A Twenty-Year Surveillance Study.

We retrospectively analyzed the antimicrobial data of Enterobacter spp. strains isolated from hospitalized subjects and outpatients over 20 years (2000-2019). A total of 2277 non-duplicate Enterobacter spp. isolates, 1037 from outpatients (45%) and 1240 from hospitalized subjects (55%), were retrieved. Most of samples are infections of the urinary tract. Considering Enterobacter aerogenes, now classified as Klebsiella aerogenes, and Enterobacter cloacae, representing more than 90% of all isolates, except for aminoglycosides and fluroquinolones, which showed significant antibiotic decreasing trends (p < 0.01), none of the other antimicrobial agents tested showed significant changes in both groups (p > 0.05). Conversely, there was a significant increasing resistance trend for fosfomycin (p < 0.01), among both community and hospital-related subjects, most probably owing to uncontrolled and improper usage. Surveillance studies on antibiotic resistance at the local and regional level are required to detect new resistance mechanisms, reduce inappropriate antimicrobial consumption, and increase the focus on antimicrobial stewardship.

Isolation and Molecular Identification of Enterobacter cloacae from Pregnant Women with Urinary Tract Infections in Biu, North Eastern Nigeria, Borno State

Urinary Tract Infections (UTIs) represent a significant, widespread, and expensive public health issue owing to their high frequency and the associated health challenges on pregnant women. This study aimed to isolate, molecularly identify Enterobacter cloacae strains from pregnant women with Urinary tract infections in some General Hospitals in Biu, Borno State. A total of 250 vaginal swabs from pregnant women who attended Antenatal Care Unit whose age ranges from 15 to 45 years were recruited using a well-designed questionnaire by random sampling technique. Gram-staining, culture, and wet microscopy were employed to check the presence of bacterial pathogens in the samples. Polymerase Chain Reaction was carried out to further validate the results. The degree of relatedness of the organism was determined using NCBI software. The result revealed that 167 representing 66.8% of the total samples studied showed significant bacterial growth. The ascending order of the percentage of the bacterial agents isolated include: Enterobacter cloacae 47 (28.1%), Klebsiella pneumoniae 36 (21.6%), Escherichia coli 23 (13.8%), Proteus mirabillis 19 (11.4%), Staphylococcus epidermis 15 (9.0%), Staphylococcus aureus14 (8.3%) and Pseudomonas aeruginosa 13 (7.8%). The result of the demographic factors showed that 17 (36.2%) of pregnant women infected within the age of 15–20 years had Enterobacter cloacae. Illiterate pregnant women were 19 (40.4%), unemployed pregnant women were 29 (61.7%), those in their third trimester (27-40 weeks) were 24 (51.1%), women who had 7 and above pregnancies 25 (53.2%), symptomatic and asymptomatic pregnant women within the age of 15–20 years 19 (40.4%), and rural dwellers 33 (70.2%). This study also demonstrated culture technique as the best biochemical method for isolation of bacterial agents compared to wet mount and microscopic Gram-staining isolation method. It can be concluded that there is a 66.8% prevalence of Enterobacter cloacae in Biu LGA of Borno State.