Materials and Methods: Actinobacteria were isolated from soil samples of the desert areas of Qom province, Iran. Biochemical and molecular tests of 16S rRNA were used to identify Actinobacteria isolates. The produced biosurfactant was investigated by methods of hemolysis, oil droplet destruction, lipase production, oil expansion, emulsifying activity, and surface tension reduction measurement. The structure of biosurfactant was investigated by Fourier transform infrared spectroscopy (FTIR) analysis, and its effect on bfp gene expression was measured. Also, isolates of A. hydrophila were obtained from stool samples of children referred to Hazrat Masoumeh Hospital in Qom from May 2022 to March 2023. Then, the effect of a biosurfactant isolated from Actinobacteria on the bfp gene expression of A. hydrophila isolates was measured by RT-PCR.

Results: Based on sequencing data, the Streptomyces genus with the ability to produce biosurfactant was isolated from the soil of the studied area, which could reduce the expression of the bfp gene after treatment with biosurfactant in clinical isolates of A. hydrophila.

Discussion: The biosurfactant-producing isolates were identified as Streptomyces spp. The results indicated that the biosurfactant significantly decreased bfp gene expression in A. hydrophila. This emphasizes the potential of biosurfactants to eliminate microorganisms by reducing virulence gene expression, inhibiting biofilm formation, demonstrating antimicrobial activity, and improving emulsification. The study supports the idea that biosurfactants can interfere with bacterial mechanisms that cause disease, such as biofilm formation, which is critical for pathogen persistence and resistance. Previous research also confirms the antipathogenic activity of natural isolates against A. hydrophila.

Conclusion: The findings of the present study show that the desert soils of Qom province are a potential area for finding actinobacterial isolates with the ability to produce biosurfactants and influence the expression of pathogenic genes of clinical isolates of A. hydrophila.

Methods: This analytical report is the first of its kind in this domain and provides a comprehensive overview of the available patents associated with the conservation of horseshoe crabs. Patents associated with horseshoe crabs were searched in PatSeer and the data analysed and filtered, based on relevance.

Results: The analysis is based on an extensive dataset (413) that describes technology for conservation of these living fossils, with a focus on recombinant proteins that can be a viable alternative to the mass utilization of the horseshoe crabs for the extraction of limulus amoebocyte lysate. Other technological advances which advocate cell-free hemolymph production and the use of artificial baits to replace the traditional grassroot practices, procedures related to efficient breeding, growth, hatching and release from artificial culture systems can go a long way in the conservation of these living fossils.

Discussion: The technologies and innovation reveal possible means of reducing the dependence on live animals through non-invasive methods. Novel interventions such as recombinant Factor C for endotoxin detection provide promising alternatives to conventional methods. Additionally, technological advances in aquaculture protocols provide strategies that allow for the conservation and artificial breeding of the horseshoe crab.

Conclusion: Transitioning to recombinant Factor C, a ban on horseshoe crab baits, diversity mapping through genetic markers and artificial breeding techniques are some of the measures that can be manifested at the policy level to enhance conservation efforts.

Methods: Total flavonoid and phenolic contents were determined by aluminum chloride and Folin-Ciocalteu techniques, respectively. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2- azino-bis (3-ethylbenzothiazoline-6-sulphuric acid (ABTS) approaches were used for assessing the scavenging of free radicals. Employing standard methods, the preliminary phytochemical activity of the extract was assessed, and its reducing power was tested in the presence of potassium hexacyanoferrate.

Results: A maximum concentration of 58.88 ± 0.32 of gallic acid was identified per gram of methanolic extract as phenolic content. Quercetin, at 85.51 ± 0.40 mg/gram, had the most flavonoid content. The strongest reducing power output measured was 1.968 ± 0.01 g. In comparison, the IC50 values for DPPH and ABTS assays were 162.79 ± 0.24 and 39.75 ± 0.20 μg/ml, respectively. Thus, this plant differs from others in that it has potent antioxidant capabilities.

Conclusion: The methanolic crude extract of D. hatagirea and the estimated contents of phenols and flavonoids exhibited potent antioxidant action, particularly against DPPH and ABTS assays. Due to its relatively high phenolic and flavonoid contents, this plant is an exciting option for treating diseases. Therefore, secondary metabolites can be found and employed as low-resistance, multi-target antioxidant drugs for treating a variety of cancers, cardiovascular and neurodegenerative disorders, as well as other chronic diseases that are resistant to existing antibiotics. Our plant will be patented for its pharmacological applications.

Materials and Methods: The synthesis of CuO nanoparticles was performed using the aqueous extract of the leaves of Ephedra major plant .The synthesized nanoparticles were evaluated by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). The well diffusion method investigated the antimicrobial activity of CuO-NPs synthesized against Pseudomonas aeruginosa. Then, the MIC and MBC of the synthesized nanoparticles were determined in 96-well microplates with different concentrations of CuO-NPs, aqueous extract, and chloramphenicol. The inhibition of Pseudomonas aeruginosa biofilm was investigated by staining with 1% crystal violet.

Results: The results of the UV-Vis analysis showed that the absorption at the wavelength of 385 nm was the highest, which confirmed the formation of CuO-NPs. SEM and EDX results indicated that the nanoparticles formed in a spherical shape with an average size of 30 to 80 nm. Also, EDX analysis showed the presence of copper, carbon, and oxygen elements in nanoparticles. The CuO-PNs at the concentration of 2000 μg/ml exhibited a significant inhibitory effect against P. aeruginosa. Also, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of nanoparticles were 312 μg/ml. In addition, the results showed that CuO-NPs have an effect in inhibiting biofilm formation. The inhibitory effect against biofilm was greater with increasing concentration.

Conclusion: The results of this study prove that CuO-NPs synthesized from the aqueous extract of Ephedra major plant can be used as an effective option in treating infections caused by P. aeruginosa and a new patent.]]> https://www.benthamscience.com/article/1500492026-05-12 Background: Biofilm production is a key factor in the development of antibiotic resistance in multidrug-resistant Klebsiella pneumoniae (K. pneumoniae), a significant contributor to healthcare-associated infections (HAIs). Kaempferol, a flavonoid, is widely recognized for its ability to combat various microorganisms.

Aim: Our goal is to assess the impact of kaempferol on K. pneumoniae biofilms by determining the level of gene expression for the biofilm-forming genes.

Methods: Fifty K. pneumoniae isolates were studied. Different doses of kaempferol with a concentration range of 0.04 to 100% in Luria Bertani broth (LB) medium were incubated at 37°C for 24 h with forty-three K. pneumoniae strong and intermediate biofilm producers. The minimum inhibitory concentration (MIC) of kaempferol was determined. Molecular detection of the biofilm-forming genes (mrkA, pgaA, wbbM, and wzm) was performed on all isolates before and after kaempferol treatment at 0.5 x MIC.

Results: Seven isolates out of 50 (14%) exhibited weak biofilm formation ability, 6 out of 50 (12%) were moderate producers, and 37 out of 50 (74%) were strong producers. The MIC values of kaempferol for K. pneumoniae ranged from 50% to 6.25% (p = 0.0003). The levels of expression of the studied genes were slightly decreased after treatment compared with their corresponding values before treatment.

Discussion: Kaempferol has shown potential in disrupting biofilms by inhibiting key genes (mrkA, pgaA, wbbM, wzm) involved in adhesion and biofilm matrix synthesis, although its effect is moderate. In vitro testing revealed that kaempferol inhibits biofilm formation at varying concentrations depending on the bacterial strain, with gene expression downregulation indicating its interference in biofilm-related pathways. Patent related to this topic has been mentioned along the text.

Conclusion: Based on current knowledge, few research studies have investigated the impact of kaempferol on K. pneumoniae biofilms. Our results show that its effect on the biofilms of this bacterium is moderate to weak. Further research is necessary to determine potential synergies with other treatments.]]> https://www.benthamscience.com/article/1555792026-05-12