Cloacae Bacteria Research Articles

Purification of antibacterial attacins induced in Hyalophora cecropia pupae immunized with Enterobacter cloacae C7‐501 using ion‐exchange chromatography, hydrophobic interaction chromatography, and Rotofor® isoelectric focusing

AbstractHyalophora cecropia pupae were infected by Enterobacter cloacae C7‐501 to induce antibacterial attacins for purification. The induction of attacins in immunized pupae was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE). Ion‐exchange chromatography (IEC), hydrophobic interaction chromatography (HIC), and Rotofor® isoelectric focusing (ISEF) were applied to isolate attacins from the hemolymph. IEC separated attacins from most hemolymph proteins, but the fractions containing attacins also had other proteins of 20 and 64 kDa in length. In IEC, attacin was eluted with ~0.2 M NaCl. The best conditions for IEC were pH 9, flow rate of 2 mL/min, with step elution (0.025, 0.05, 0.075, 0.1, 0.2, 0.4 and 1.0 M NaCl). In HIC, most other proteins were eluted with the ammonium persulfate treatment. HIC isolated attacin proteins under hydrophobic conditions, at ~50% EtOH. However, the fraction with attacins also contained other proteins. The Rotofor® ISEF produced fractions containing attacins at isoelectric points ranging between 5.7 and 8.3. However, non‐specific proteins were detected in the fraction samples, and the recovery of attacins was low. The purification efficiency of ISEF was lower than IEC and HIC. In this study, the expression of attacins was induced in H. cecropia pupae infected with E. cloacae C7‐501, and attacins could be purified by IEC and ISEF. Overall, IEC provided better separation of attacins from the hemolymph of H. cecropia pupae immunized with E. cloacae bacteria than HIC and Rotofor® ISEF.

Impacts of essential oil and extracts obtained from coriander cultivars (Coriandrum sativum L.) on important some pathogenic bacteria

In this study, the antibacterial activity of the essential oil obtained from the seeds of two coriander (Coriandrum sativum L.) cultivars (Gürbüz and Arslan) and extracts obtained from the seeds, leaves and stems with methanol and ethanol against 7 Gram-positive (Escherichia coli ATCC®25922, Pseudomonas aeruginosa ATCC®9027, Salmonella typhimurium ATCC®14028, Serratia marcescens ATCC®13880, Proteus vulgaris ATCC®6380, Enterobacter cloacae ATCC®13047, and Klebsiella pneumoniae ATCC®4352), and 1 Gram-negative bacteria (Staphylococcus aureus ATCC®6538) were evaluated. Summer sowing was carried out for the supply of herbal materials and the plants were grown in Yozgat / TURKEY climatic conditions. Linalool (average 74%) has been noted as the main component in essential oils, and γ-terpinene, geraniol and camphor have been identified as other important components. It has been determined that essential oils exhibit varying levels of activity against S. aureus, E. coli, S. tyhimurium, S. marcescens, P. vulgaris, and E. cloacae bacteria in this study.

Distribution of nitrogen-transforming bacteria in an artificial reservoir populated with aquatic higher plants

This article examines the distribution of nitrogen-transforming bacteria in an artificial reservoir (pond) populated with aquatic higher plants of common reed (Phragmites australis) and cattail (Typha) for treating swine wastewater. In the pond occupied by Phragmites australis, 7 strains of ammonium oxidising and 14 denitrifying bacteria were identified, while, in the pond occupied by Typha, 6 strains of ammonium oxidising and 19 denitrifying bacteria were distinguished. A comparative analysis of bacterial count at various sampling points revealed their decrease along the artificial pond. Most of the bacteria strains oxidising ammonium were identified in the samples collected from the surface layer of the pond, while denitrifying bacteria dominated the bottom layer. The isolated microorganisms identified by 16S rRNA sequencing belonged to the genus Pantoea, Enterobacter and Bacillus. An artificial pond having aquatic higher plants is characterised by a diverse microbiota, whose composition strongly depends on the wastewater source rather than on the cultivated plant species. The ammonia transformation and denitrification capacity of isolated bacterial strains was determined. The highest conversion efficiency of ammonia (up to 56%) was observed for Enterobacter cloacae bacteria isolated from the surface layer of the pond. Denitrifying bacteria sampled from the bottom layer allowed for the reduction in nitrate content from 20 to 10 mg/L in 72 h. The obtained results confirm the role of bacteria in the treatment of wastewater against nitrogen-containing pollutants, with the average efficiency of removal of inorganic nitrogen compounds being 50%.

Evaluation of anti-malaria potency of wild and genetically modified Enterobacter cloacae expressing effector proteins in Anopheles stephensi

BackgroundMalaria is one of the most lethal infectious diseases in tropical and subtropical areas of the world. Paratransgenesis using symbiotic bacteria offers a sustainable and environmentally friendly strategy to combat this disease. In the study reported here, we evaluated the disruption of malaria transmission in the Anopheles stephensi-Plasmodium berghei assemblage using the wild-type (WT) and three modified strains of the insect gut bacterium, Enterobacter cloacae.MethodsThe assay was carried out using the E. cloacae dissolvens WT and three engineered strains (expressing green fluorescent protein-defensin (GFP-D), scorpine-HasA (S-HasA) and HasA only, respectively). Cotton wool soaked in a solution of 5% (wt/vol) fructose + red dye (1/50 ml) laced with one of the bacterial strains (1 × 109cells/ml) was placed overnight in cages containing female An. stephensi mosquitoes (age: 3–5 days). Each group of sugar-fed mosquitoes was then starved for 4–6 h, following which time they were allowed to blood-feed on P. berghei–infected mice for 20 min in the dark at 17–20 °C. The blood-fed mosquitoes were kept at 19 ± 1 °C and 80 ± 5% relative humidity, and parasite infection was measured by midgut dissection and oocyst counting 10 days post-infection (dpi).ResultsExposure to both WT and genetically modified E. cloacae dissolvens strains significantly (P < 0.0001) disrupted P. berghei development in the midgut of An. stephensi, in comparison with the control group. The mean parasite inhibition of E. cloacaeWT, E. cloacaeHasA, E. cloacaeS−HasA and E. cloacaeGFP−D was measured as 72, 86, 92.5 and 92.8 respectively.ConclusionsThe WT and modified strains of E. cloacae have the potential to abolish oocyst development by providing a physical barrier or through the excretion of intrinsic effector molecules. These findings reinforce the case for the use of either WT or genetically modified strains of E. cloacae bacteria as a powerful tool to combat malaria.Graphical

Study the pathogenicity of the bacteria associated with Periplanta americana cockroach on Gelleria mollonella. L worm larva

In this study, the bacterial genera associated with the Periplaneta americana cockroach were isolated from the residential areas and nurseries in Baghdad city with concentrations 10-2, 10-4, 10-6 and their effect on the larvae of Gelleria mollonella. L worm was studied in the laboratory by food spraying method. Conclusion: The results showed that the highest percentage of larvae mortality was at the concentration 10-2 (33.33%) for both Acinetobacter lwoffii and Enterobacter cloacae bacteria. While the highest rate of larvae mortality was at the concentration 10-4 (33.33%) for each of Pseudomonas oleovorans, Acinetobacter baumannii, Acinetobacter lwoffii and Klebsiella pneumoniae, whereas the concentration 10-6 showed the highest mortality rate (50%) of Pseudomonas oleovorans and Klebsiellaella bacteria. Results of the study showed predominance of the effect of Pseudomonas oleovorans and Klebsiella pneumonia on the mortality rate of the larvae of the great Gelleria mellonell L. worm as it was 50%, while the effect of Enterobacter cloacae and Pseudomonas oleovorans yielded the highest mortality rate of 33.33% at each of the concentrations 10-2,10-4 10-6 whereas the results showed that the concentration 10-2 for each of Proteus mirabilis, Escherichia coli, and Enterobacter aerogenes had no effect on larvae mortality.

ABILITY OF AMILOLYTIC BACTERIA (Bacillus paramycoides and Enterobacter cloacae) IN DEGRADING ORGANIC MATERIALS OF MANGROVE LITTLE

This study aims to find out that Bacillus paramycoides and Enterobacter cloacae bacteria can produce amylase enzymes and have the ability to degrade organic matter, especially mangrove litter. From this study it was found that the optimal growth of B.paramycoides and E. cloacae bacteria occurred at 12th hour. The results of measurements and calculations of absorbance values ​​at 630 10.238 x 108 cells/mL (B. paramycoides) and 12.030 x 108 cells/mL (E. cloacae) using the spectrophotometric method. Meanwhile, with the TPC method at 12 hours, the number of bacterial cells was 2.08 x 108 CFU's/ml (B. paramycoides) and 2.44 x 108 CFU's/ml (E. cloacae). The ability to produce the largest amylolytic bacterial amylase enzyme also occurred at 12 hours as much as 0.306 mg/mL (B.paramycoides) with an increase of 0.046 mg/mL and 0.243 mg/mL (E. cloacae) with an increase of 0.028 mg/mL. The bacteria that have the highest amylase enzyme ability is E.cloacae as evidenced by the diameter of the clear zone of 10.10 mm. Testing the ability of amylolytic bacteria in degrading mangrove litter was carried out by adding amylase enzyme as much as 0%, 50% and 100%. Amylolytic bacteria can degrade organic matter by hydrolyzing starch contained in mangrove litter. The most degraded starch content was in the 100% enzyme treatment, which was 1.021 mg/mL (B. paramycoides) and 1.189 mg/mL (E.cloacae).

Daya Hambat Kayu Manis (Cinnamomum burmanni) terhadap Pertumbuhan Bakteri Kultur Darah Widal Positif Anggota Familia Enterobacteriaceae

Cinnamon has a high activity of antibacterial. Still, the antibacterial effect toward the bacteria on the positive Widal blood culture of Enterobacteriaceae family members was not clear yet. This research aimed to find out the inhibition of cinnamon on the growth of bacterium on the positive Widal blood culture of Enterobacteriaceae family members. The method used in the research was experimental research which was using the draw well method and using Salmonella typhi, Escherichia coli, Klebsiella pneumonia, Serratia marcescens, and Enterobacter cloacae bacteria samples. Cinnamon as a test solution with a 2%, 4%, 6%, 8% and 10% concentrate. The research result was showed the inhibiting zone was able to form on the Nutrient Agar Plate (NAP) on growth of the Enterobacteriaceae, 10% concentrate that high sensitive showed to S. typhi, Ser. marcescens, E. cloacae, E.coli, and Kleb. pneumonia squent was 12 mm, 11 mm, 11 mm, 10 mm, and 8,5 mm. At the same time, 8%, 6%, 4%, and 2% concentrate of cinnamon could not perform inhibition zone 100 µl of the test solution with chloramphenicol as a positive control.

The Effectiveness of Wild Honey In Inhibiting The Growth of Bacterium on The Positive Widal Blood Culture of Enterobacteriaceae Familia

Honey is a product produced by bees which is believed has many benefits in the medical field. Wild honeys are more natural than livestock honey, and it has high antimicroba activities, but the effect of the antimicroba towards the bacteria on the positive widal blood culture of Enterobacteriaceae familia member was not clear yet. This research was to find out The effectiveness of wild honey in inhibiting the growth of bacteria on the positive widal blood culture of Enterobacteriaceae familia member. Method in this research was an experimental research which was using combination of diffusion and draw well method using Klebsiella pneumonia, Esherichia coli, Salmonella typhi, Serratia marcescens, and Enterobacter cloacae bacteria sample. Wild honey as a test solution with a 25%, 40%, 55%, 70%, 85% and 100% concentrate. The result is that wild honey is effective to inhibit the growth of the Enterobacteriaceae bacteria familia, the inhibition zone which is performed on the 70% Nutrien Agar Plate (NAP) concentrate medium is a minimal concentrate that sensitive to all bacteria with the inhibition zone similar to Kloramfinekol antibiotics. S. typhi is sensitive to all wild honey concentrate within 26,5mm, 29mm, 31,5mm, 32mm, 35mm, and 38mm inhibition zone. Wild honey on E.coli and Serratia marescens is sensitive in all concentrate except on the 25% concentrate. On the Klebsiella pneumonia the wild honey is sensitive on the 70%, 85%, and 100% concentrate while on the Enterobacter cloacae the wild honey is sensitive on 55%, 70%, 85% and 100% concentrate. Conclusion : wild honey is effective to inhibit bacteria growth on the positive widal blood culture of Enterobacteriaceae familia member and there are effect of various wild honey concentrate with the growth of Enterobacteriaceae bacteria which the higher concentrate of the wild honey, the greater inhibition potency towards the bacteria.

514 Optimal Bacterial Surveillance Protocol for Duodenoscopes and Linear Echoendoscopes: A Comparison of Two Approaches

INTRODUCTION: The complex design of the elevator mechanism in duodenoscopes has been recognized as a challenge for disinfection and previously implicated as a source of clinically significant bacterial transmission. Our group has also determined the potential for harboring bacteria in elevator-containing linear echoendoscopes (1). Despite increased awareness, there continues to be a lack of definitive guidelines for routine bacterial culturing protocols for these endoscopes. Our study aims to provide a bacterial culture protocol that is cost-effective, efficient, and accurate. METHODS: Consecutive duodenoscopes and linear echoendoscopes (Olympus; Center Valley, PA) used at a tertiary care center were reprocessed following hospital and manufacturer protocol and dried using an automatic machine. The following morning, these endoscopes were sequentially cultured for surveillance using two different protocols. The brush protocol consisted of suctioning and dispensing sterile water through the endoscope multiple times prior to brushing eight different components of the endoscope, costs $53.40 per culture, and takes an average of 15 minutes. The swab protocol consisted of using a flocked swab to sample above, below, and around the elevator mechanism while it was in mid-position, costs $32.04 per culture and takes 1 minute to perform. RESULTS: A total of 292 primary cultures were collected from 17 duodenoscopes and 8 linear echoendoscopes. Of these, 146 cultures were gathered using the brush method and 146 cultures using the swab method. No endoscopes tested positive using the brush method. Two cultures (1.4%) tested positive for Enterobacter cloacae bacteria using the swab method (one duodenoscope and one linear echoendoscope). CONCLUSION: In our pilot study data, the swab method appeared to be superior to the brush method when evaluating the presence of residual bacteria found on duodenoscopes and linear echoendoscopes. The swab method not only detected two positive cultures not found on the brush method. Furthermore, the brush method poses a risk for environmental contamination due to the increased duration of sampling. Based on our prior published data, the swab method could potentially save our center $26,529 and 290 nursing hours annually, supporting the use of the flocked swab methods. Further prospective studies in larger sample sizes are needed to validate optimal surveillance protocols for elevator containing endoscopes.

Experimental Evaluation of DBD Reactor Applied to Bacterial Inactivation in Water Flowing Continuously

In this paper, we present the evaluation of a DBD reactor that generates an energy per pulse of 65.3 mJ in order to inactivate E. coli ATCC 8739 at 103, 106, and 108 CFU/mL order concentrations and E. cloacae CDBB-B-963 at 103 and 107 CFU/mL order concentrations, in a volume of water flowing continuously. Particularly, experimentation is focused in 500 mL, under atmospheric pressure. The DBD reactor was coupled to a peristaltic pump type, a cooling jacket, containers for water in treatment and cooling fluid, and systems to monitor the experimental conditions. The implementation of the experiments was carried out at three different fluid flow rates: 2.0±0.1, 3.5±0.1, and 4.4±0.1 mL/s; in each of them the time required to carry out a cycle of treatment corresponded to 256±7, 143 ± 2, and 113 ± 1 s, respectively. During the DBD application, water flowed continuously at recirculation mode, promoting the continuous oxygen injection at 8.3 mL/s. In the three established fluid flow rates, the efficacy of treatment reached 100% in inactivation of E. coli bacteria at 103 and 106 order concentrations at the end of the first cycle of treatment. At 108 CFU/mL order concentration, complete inactivation of E. coli bacteria was achieved at the end of the fourth cycle of treatment. Meanwhile, E. cloacae bacteria showed an inactivation of 100% at the end of the first cycle of treatment at 103 order concentration; when the order concentration increased to 107, the efficacy of the process depended on the fluid flow rate, at 3.5 ± 0.1 and 2.0 ± 0.1 mL/s survival was 0% at the end of the first cycle of treatment; nevertheless, at the highest (4.4 ± 0.1 mL/s), an additional cycle was required.

Surface-Mediated Chromate-Resistant Mechanism of Enterobacter Cloacae Bacteria Investigated by Atomic Force Microscopy

The Enterobacter cloacae CYS-25 strain isolated from a chromate plant shows a strong capability for chromate resistance instead of chromate reduction in aerobic conditions. In this study, atomic force microscopy (AFM) was used for studying the morphology characteristics of bacterial properties during the chromate resistance process. The average length of E. cloacae bacteria in the stationary phase is about 2.3 +/- 0.6 microm, while under the stimulation of 400 mg/L CrO42-, the length of bacteria increases to 3.2 +/- 0.7 microm. Height and phase images showed that, with the addition of CrO42-, the smooth surface of bacteria changed into one with discontinuous features with characteristic dimension of 40-200 nm. Analysis reveals that these compact convex patches are organic components stimulated by CrO42-. A chromate resistance mechanism relating to the overexpression of extracellular biologic components for preventing the permeability of CrO42- into the cell is proposed as the survival strategy of E. cloacae in chromate situation.

Isolation of mannose-binding C-type lectin from Heliothis virescens pupae

A mannose-binding C-type lectin (MBL) was isolated by affinity chromatography from Heliothis virescens immune pupal hemolymph. The immune pupal hemolymph was obtained after bacterial injection of live Enterobacter cloacae bacteria. MBL in mammals acts as an opsonin for phagocytosis and activates the lectin complement pathway of the innate immune response, which leads to killing of gram-negative bacteria and enveloped viruses. The affinity-purified and reduced pupal MBL showed a single band of 36 kDa by SDS–PAGE (12% gel). A dot-immunoblot ELISA (using guinea pig anti-MBL IgG as primary antibody) demonstrated specificity of the antibody for the affinity-purified pupal MBL. The immune pupal hemolymph contained 21 μg of MBL per ml of hemolymph. The amino acid composition of the purified pupal MBL was determined with high amounts of arginine and histidine detected. The presence of MBL in insect pupae has not before been reported and could be important in pupal innate immunity to bacterial infection.