Detection of Virulence Gene Profiles of Multi-Drug Resistance (MDR) in Pseudomonas aeruginosa Bacteria in Human

The spread of opportunistic bacteria Pseudomonas aeruginosa in hospitals correlated with several diseases to humans. This bacterium showed high resistance to many antibiotics due to their indiscriminate use, as well as mutations in the pathogenic genes of Pseudomonas aeruginosa, which are difficult to treat. Pseudomonas aeruginosa is transmitted to humans in hospitals as being special nosocomial bacteria through ventilators, also through food and water contaminated with these bacteria, which causes muscle fatigue, vomiting and nausea. It also weakens the immune system and is more severe in people who suffer from weak immunity, old age and children, and can lead to death. Therefore, scanning virulence genes in Pseudomonas aeruginosa helps understand pathogenesis mechanisms. The main objective of this study was to detect three pathogenic genes in these bacteria: ExoS, Apr, and Pich. Bacteriological samples were collected and detected by the Multiplex PCR mechanism. Results showed that the sizes of genes were: 444bp, 1017bp, and 608bp. The study of the reaction of multiplex polymers to detect genes (ExoS, Apr, Pich) in 32 colonies was provided by Al-Ahli Hospital in Hebron, the percentage of each gene was 95%. In addition to 12 samples provided by the Al-Istishari Hospital in the city of Ramallah, and the percentage of the presence of each gene in these samples 100%. In the antibiotic examination of Pseudomonas aeruginosa of 44 samples, the bacteria had antibiotic resistance, 43% and 31% of the bacterial strains were resistant to Gentamicin and Aztreonam, 27% of which were resistant to Meropenem, Ceftazidime and Ciprofloxacin, 18% of which were resistant to tazobactam and 22.7% of which were resistant to Amikacin. The minor objective was to identify MDR Pseudomonas aeruginosa from clinical isolates. Isolates were evaluated for their antimicrobial susceptibility to seven antibiotics, Meropenem, Ceftazidime, Amikacin, Azteronam, Ciprofloxacin, Gentamicin and Piperacillin-tazobactam. The results also provided a clear picture of bacterial resistance MDR to these antibiotics by 45.45%

Isolation of bacteria that are able to digest keratin as an alternative to harmful chemicals used in tanning industries

Abstract:Macroccocus is a bacteria that was isolated from a soil sample containing decaying hair and It was found to contain keratinase enzyme due to its high ability to digest keratin protein. The isolated bacteria by this study can be used as an alternative to harmful chemicals tanning processes represents a safe and, clean technology that achieves economic feasibility in the process of skin soaking and hair removal. Background: Leather tanning is considered one of the ancient professions that Hebron has been known for hundreds of years which is the only source of raw materials for Palestinian factories thatdepends on this commodity. In the past, the industry relied on environmentally friendly natural materials in tanning, but today they have become a source of pollution of soil and groundwater, especially after the entry of chemicals in this craft, such as chromium and some acids that infect humans and destroy the Palestinian environment because of the toxic gases that result from the process of soaking and hair removal. Objectives: 1. To Isolate keratinolytic bacteria from soil sample 2. To test the bacterial effectiveness for hair degradation 3. To identify and characterize of bacteria by sequencing tool. Methods: Soil sample containing decaying hair was collected from the industrial area in Hebron city and processed in the lab, bacteria were isolated by using spreading and streaking plate technique, their ability for hair degradation was tested by incubating bacterial colony, hair and water for several days, after the bacterial ability for hair degradation appeared bacterial isolate was identified by sequencing tool and the type of bacteria was determined by bioinformatics tool. Results: Isolated a bacterial strain that is able to digest keratin and was tested on several samples and proved to be effective in degrading hair. It has been welcomed by the Modern Leather Tanning Company (MLTC-Hebron), which has cooperated with us in this project. The strain was genotyped and appeared to belong to macroccocus, this is the first report to find an industrial value for such bacteria which is also safe for human and the environment. Conclusions: Among microbial isolate screened for keratin digestion by using soil sample, a newly isolated Macroccocusequipercicuswas found with a greater ability for hair degradation, the use of such bacteria as an alternative to harmful chemicals tanning processes represents a safe and, clean technology that achieves economic feasibility in the process of skin soaking and hair removal. but in the same time, the full sequence of these strains of bacteria is unknown and not significantly studied. Therefore, additional researches have to be done for characterization of this type of bacteri

Traitement multimodal du portage intestinal de bactéries multirésistantes avec des probiotiques et des prébiotiques

The intestine is the main reservoir of multidrug-resistant Enterobacterales (MDRE), and there is an urgent need to identify treatments for intestinal colonization by MDRE. Our objectives were to test innovative treatments for digestive colonization by ESBL Escherichia coli (ESBL-Ec) in an experimental mouse model of intestinal dysbiosis induced by amoxicillin, and to predict the efficacy of treatments from the composition of the microbiota before treatment. We obtained a strong in vitro activity of new strains of B. subtilis against ESBL-Ec. However, these strains were not effective in vivo. We then showed a temporary in vivo activity of commensal E. coli. In addition, treatments with Akkermansia muciniphila and inulin were significantly effective in mice, but only when combined with pantoprazole. Pantoprazole alone showed no effect on ESBL-Ec colonization, but alleviated the effect of amoxicillin on the intestinal microbiota. qPCR and metagenomic analysis identified Adlercreutzia caecimuris and A. muris among other taxa as predictors of efficacy of the inulin/pantoprazole combination. We showed that, administered as a treatment, they significantly reduced fecal titers of ESBL-Ec. These taxa could have an additive effect with other tested treatments. It would be interesting to test such new symbiotic mixtures to reduce intestinal colonization by ESBL-Ec. In addition, the strong beta-lactamase activity detected in feces before ESBL-Ec inoculation was remarkably associated with subsequent decolonization, likely by decreasing amoxicillin fecal concentration and promoting rich gut microbiota. This study therefore opens up new avenues for the treatment of intestinal colonization by multi-drug resistant Enterobacterales .L'intestin est le principal réservoir d'entérobactéries multirésistantes (MDRE), et il est urgent d'identifier des traitements de la colonisation intestinale par MDRE. Nos objectifs étaient de tester des traitements innovants de la colonisation digestive par un Escherichia coli producteur de BLSE (EcBLSE) dans un modèle expérimental murin de dysbiose intestinale induite par l'amoxicilline, et de prédire l'efficacité des traitements à partir de la composition du microbiote avant traitement. Dans ce travail, nous avons obtenu une forte activité in vitro de nouvelles souches de B. subtilis contre EcBLSE. Cependant ces souches se sont révélées inactives in vivo. Nous avons ensuite montré une activité temporaire in vivo de souches d’E. coli commensales. En outre, les traitements par Akkermansia muciniphila et inuline étaient efficaces chez la souris, mais uniquement lorsqu'ils étaient associés au pantoprazole. Le pantoprazole seul n'a montré aucun effet sur la colonisation par notre EcBLSE, mais il réduisait la dysbiose induite par l’amoxicilline. Les analyses par qPCR et shotgun sequencing ont identifié Adlercreutzia caecimuris et A. muris parmi d'autres taxons comme prédicteurs de l'efficacité de la combinaison inuline/pantoprazole. Nous avons pu montrer qu’utilisés comme traitement préventif, ils réduisaient significativement les titres fécaux d’EcBLSE Ces taxons pourraient avoir un effet additif sur nos traitements. Il sera donc intéressant de tester de nouveaux mélanges symbiotiques pour diminuer la colonisation intestinale par les EcBLSE. De plus, la forte activité bêta-lactamase détectée dans les fèces avant inoculation d’EcBLSE était remarquablement corrélée à la décolonisation ultérieure, probablement en diminuant la concentration fécale d’amoxicilline et en réduisant la dysbiose induite par l’amoxicilline. Cette étude ouvre donc de nouvelles voies pour le traitement de la colonisation intestinale par les entérobactéries multirésistantes

Traitement multimodal du portage intestinal de bactéries multirésistantes avec des probiotiques et des prébiotiques

L'intestin est le principal réservoir d'entérobactéries multirésistantes (MDRE), et il est urgent d'identifier des traitements de la colonisation intestinale par MDRE. Nos objectifs étaient de tester des traitements innovants de la colonisation digestive par un Escherichia coli producteur de BLSE (EcBLSE) dans un modèle expérimental murin de dysbiose intestinale induite par l'amoxicilline, et de prédire l'efficacité des traitements à partir de la composition du microbiote avant traitement. Dans ce travail, nous avons obtenu une forte activité in vitro de nouvelles souches de B. subtilis contre EcBLSE. Cependant ces souches se sont révélées inactives in vivo. Nous avons ensuite montré une activité temporaire in vivo de souches d’E. coli commensales. En outre, les traitements par Akkermansia muciniphila et inuline étaient efficaces chez la souris, mais uniquement lorsqu'ils étaient associés au pantoprazole. Le pantoprazole seul n'a montré aucun effet sur la colonisation par notre EcBLSE, mais il réduisait la dysbiose induite par l’amoxicilline. Les analyses par qPCR et shotgun sequencing ont identifié Adlercreutzia caecimuris et A. muris parmi d'autres taxons comme prédicteurs de l'efficacité de la combinaison inuline/pantoprazole. Nous avons pu montrer qu’utilisés comme traitement préventif, ils réduisaient significativement les titres fécaux d’EcBLSE Ces taxons pourraient avoir un effet additif sur nos traitements. Il sera donc intéressant de tester de nouveaux mélanges symbiotiques pour diminuer la colonisation intestinale par les EcBLSE. De plus, la forte activité bêta-lactamase détectée dans les fèces avant inoculation d’EcBLSE était remarquablement corrélée à la décolonisation ultérieure, probablement en diminuant la concentration fécale d’amoxicilline et en réduisant la dysbiose induite par l’amoxicilline. Cette étude ouvre donc de nouvelles voies pour le traitement de la colonisation intestinale par les entérobactéries multirésistantes.The intestine is the main reservoir of multidrug-resistant Enterobacterales (MDRE), and there is an urgent need to identify treatments for intestinal colonization by MDRE. Our objectives were to test innovative treatments for digestive colonization by ESBL Escherichia coli (ESBL-Ec) in an experimental mouse model of intestinal dysbiosis induced by amoxicillin, and to predict the efficacy of treatments from the composition of the microbiota before treatment. We obtained a strong in vitro activity of new strains of B. subtilis against ESBL-Ec. However, these strains were not effective in vivo. We then showed a temporary in vivo activity of commensal E. coli. In addition, treatments with Akkermansia muciniphila and inulin were significantly effective in mice, but only when combined with pantoprazole. Pantoprazole alone showed no effect on ESBL-Ec colonization, but alleviated the effect of amoxicillin on the intestinal microbiota. qPCR and metagenomic analysis identified Adlercreutzia caecimuris and A. muris among other taxa as predictors of efficacy of the inulin/pantoprazole combination. We showed that, administered as a treatment, they significantly reduced fecal titers of ESBL-Ec. These taxa could have an additive effect with other tested treatments. It would be interesting to test such new symbiotic mixtures to reduce intestinal colonization by ESBL-Ec. In addition, the strong beta-lactamase activity detected in feces before ESBL-Ec inoculation was remarkably associated with subsequent decolonization, likely by decreasing amoxicillin fecal concentration and promoting rich gut microbiota. This study therefore opens up new avenues for the treatment of intestinal colonization by multi-drug resistant Enterobacterales

In vitro and in vivo activity of new strains of Bacillus subtilis against ESBL-producing Escherichia coli: an experimental study

International audienceAims: The gastro-intestinal tract is a major reservoir of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli. Bacillus spores may be used as probiotics to decrease digestive colonization by ESBL-E. coli. Our aim was to assess the in vitro and in vivo activity of new Bacillus strains against ESBL-E. coli.Methods and Results: We screened the in vitro activity of 50 Bacillus strains against clinical isolates of ESBL-E. coli and selected B. subtilis strains CH311 and S3B. Both strains decreased ESBL-E. coli titers by 4 log10 CFU L−1 in an in vitro model of gut content, whereas the B. subtilis CU1 strain did not. In a murine model of intestinal colonization by ESBL-E. coli, CH311 and S3B did not decrease fecal titers of ESBL-E. coli. Ten sequences of putative antimicrobial peptides were identified in the genomes of CH311 and S3B, but not in CU1.Conclusions: Two new B. subtilis strains showed strong in vitro activity against ESBL-E. coli.Significance and Impact of Study: Despite strong in vitro activities of new B. subtilis strains against ESBL-E. coli, intestinal colonisation was not altered by curative Bacillus treatment even if their spores proved to germinate in the gut. Thus, this work underlines the importance of in vivo experiments to identify efficient probiotics. The use of potential antimicrobial compounds identified by genome sequencing remains an attractive alternative to explore