Spread of ESßL-producing Escherichia coli and the anti-virulence effect of graphene nano-sheets.

Despite the studies worldwide, the prevalence of ESßL E. coli in the Iraq is still unknown. Realization of the demographic characterization of ESßL E. coli infections will assist the prevention efforts. This study aimed to isolate clinical E. coli, determine their antimicrobial susceptibility, phenotypic and genotypic detection of ESßL-producing ability, detection of some virulence-related genes, estimate the impact of graphene nano-sheets as antibacterial, and study the adherence-related gene expressions in E. coli isolates. Graphene nano-sheets were synthesized and characterized using XRD, UV, TEM, and SEM. E. coli isolates were identified using 16S rRNA. Antibiotic resistance was detected, virulence genes (blaTEM, blaSHV, BlaCTX-M-15, papC, and fimH) were screened using PCR. The antibacterial activity of graphene nano-sheets was screened using well-diffusion assay and MIC. The gene expression of adherence genes after treatment with graphene nano-sheets was evaluated using QRT-PCR. From a total of 512 identified using 16S rRNA, 359 (69.9%) were ESßL-producing E. coli. The ESßL genotypes positive were 83.56% (300/359) of E. coli isolates with the frequencies: 85% for blaCTX-M gene, 26% for blaSHV gene, and 28% for blaTEM gene. Graphene nano-sheets showed effective antibacterial activity with MIC 25 µg/ml. Furthermore, graphene nano-sheets reduced the expression of papC, and fimH genes. This study has helped us to better understand the characteristics of ESßL E. coli, their adherence gene harboring, and the potential ability of graphene nano-sheets to reduce bacterial growth, and the expression of adherence genes. Furthermore, the current study showed further step to understand the mechanisms by which graphene nano-sheets can conflict bacterial virulence and resistance.

Treatment strategy by lactoperoxidase and lactoferrin combination: Immunomodulatory and antibacterial activity against multidrug-resistant Acinetobacter baumannii.

Lactoperoxidase (Lpo) and Lactoferrin (Lf) were extracted from camel colostrum milk and purified. The antibacterial activity of the two purified proteins was estimated against 14 isolates of multidrug resistance Acinetobacter baumannii. A combination of Lpo and Lf exhibited bactericidal action against A. baumannii in vitro. A mouse model of acute A. baumannii pneumonia was improved. The injection of combined Lpo and Lf after infection leads to significant clearance of A. baumannii rates in lung as well as blood culture P < 0.05 in comparing with control. Furthermore, the results showed a significant P < 0.05 reduction in the Bronchoalveolar lavage albumin concentration, lung injury and lactate dehydrogenase activity in comparing with control. In addition, the combination of Lpo and Lf treatment induced substantial elevation of IL-4 and IL10 concentrations p < 0.0 5 that helped to prevent damage caused by the inflammatory response. We concluded that combination of Lpo and Lf had a major inhibition effect against A. baumannii in comparing with imipenem as well as their immunomodulatory activity against resistant A. baumannii was increased by a synergistic effect of them as a crude combination. This study indicated two combined proteins consider as crucial strategy for practical treatment of pneumonia in the future.

Two novel roles of buffalo milk lactoperoxidase, antibiofilm agent and immunomodulator against multidrug resistant Salmonella enterica serovar Typhi and Listeria monocytogenes.

The increasing occurrence of multidrug resistant bacteria causing bacteremia infection, constitutes a major health problem, difficult-to-treat bacteremia due to its ability to form biofilm. Buffalo milk lactoperoxidase (BMLpo) is effective and safe to use as bacteriostatic agent. The MIC of BMLpo and amikacin were used to evaluate the antibiofilm activity against resistant L. monocytogenes and S. typhi. Prophylactic effects of BMLpo against L. monocytogenes and S. typhi bacteremia in vivo have been tested and ELISA test used to evaluate serum cytokines. Significant antibiofilm activity of BMLpo observed against the highest biofilm producer isolates. Our results showed that the prophylactic effect of BMLpo in BALB/c mice bacteremic model. A significant clearance of L. monocytogenes and S. typhi, investigated in blood and different organs tissues in BMLpo-treated infected groups when compared to the non-treated groups. Further, analysis of serum cytokines levels revealed that BMLpo prophylaxis modulates their release in different way when it compared to the control. This study showed, BMLpo effects as an alternative antibiofilm agent to compact gram negative pathogens, and protects the host against bacteremia infection. Moreover, the BMLpo role as an immunomodulatory. These investigations indicated the BMLpo crucial role in the practical clinical applications.