Background: β-Lactam antibiotics have been broadly used for the treatment of Acinetobacter baumannii infections, resulting in development of β-lactam inactivating β-lactamases. Here, we described antibiotic resistance rate, prevalence of β-lactamase-encoding genes, and clonal relationships of A. baumannii strains isolated from children referred to Children’s Medical Center in Tehran, Iran, during 2019–2020. Methods: A total of 60 non-replicate A. baumannii isolates were recovered from clinical specimens of pediatric patients. Antibiotic susceptibility testing was done by the disc diffusion method. Colistin susceptibility of isolates was performed by the broth microdilution method. β-lactamase-encoding genes were characterized by PCR. The presence of ISAba1 element upstream of the several oxacillinase genes was also checked. Genetic relatedness of isolates was determined by using random amplification of polymorphic DNA (RAPD) typing. Results: The antimicrobial susceptibility tests showed that 83.3% of A. baumannii isolates were MDR, and 40% XDR. Both MDR and XDR A. baumannii isolates were susceptible to colistin. The frequency of bla_OXA-51-like, bla_OXA-23-like, bla_TEM, bla_OXA-24-like, bla_PER, bla_SHV, bla_CTX-M, bla_OXA-58-like, and bla_IMP was 100, 93.33, 60, 36.67, 28.33, 8.33, 5, 3.33, and 1.67%, respectively. Coexistence of ISAba1/bla_OXA-23-like and ISAba1/bla_OXA-51-like was observed in 65% and 85% of isolates, respectively. RAPD analysis revealed 4 common types and 2 single types of A. baumannii isolates. Conclusions: The multiple clones harboring bla_OXA-23-like, ISAba1-bla_OXA-51-like, and ISAba1-bla_OXA-23-like were responsible for the spread of A. baumannii isolates in our clinical wards. Dissemination of the well-established clones is worrisome and would become therapeutic challenges due to the possible transferring genetic elements associated with resistance.

Alpha-amylase is one of the most widely used enzymes in the starch industry. However, industrial application of soluble alpha-amylase is hampered by changes in pH and temperature (adverse effects on enzyme stability) and activity loss, leading to higher costs. Immobilization of alpha-amylase is an efficient strategy to reduce the enzyme losing and subsequently reduces costs in this regard. Alpha-amylases are immobilized by adsorption, entrapment, covalent attachment, and cross-linking. A barrier in alpha-amylase immobilization is the large size of its substrate, namely amylose and amylopectin. Most of these immobilization methods decrease the affinity of the enzyme for its substrate as well as the maximum rate of reaction (Vmax). This review aims to study different aspects of alpha-amylase including enzyme activity, applications, structure, starch, immobilization methods, and immobilization’s obstacles to improve alpha-amylase efficiency in the industry and also lowering the costs related to providing this enzyme.

Here, we described the efficacy of colistin sub-minimum inhibitory concentrations (sub-MICs) on biofilm-forming activity, host epithelial cell adherence, and invasion capacity of Acinetobacter baumannii strains collected from children admitted to the Children’s Medical Center Hospital. Biofilm formation potency of A. baumannii clinical isolates was measured using a 96-well microtiter plate assay. Distribution of biofilm-related genes, including bap , abaI , ompA , csuE , and bla _PER-1 , was detected by PCR. The mRNA expression level of ompA and csuE was measured by qPCR in the presence of ¼ and ½ MICs of colistin. A. baumannii adhesion and invasion to eukaryotic host cells were phenotypically assayed at sub-MICs of colistin. Eighty percent (56/70) and 35.7% (25/70) of A. baumannii isolates were multidrug-resistant (MDR) and extensively drug-resistant (XDR) phenotypes, respectively. The strong, moderate, and weak biofilm producers of A. baumannii were 37.1% (26/70), 32.8%, (23/70), and 22.8% (16/70), respectively. The frequencies of biofilm-associated genes were 100% for abaI , ompA , and csuE , followed by 22.8% (16/70) and 24.3% (17/70) for bap and bla _PER-1 , respectively. The downregulation of csuE and ompA expression levels was observed in the sub-MIC of colistin. In vitro cell culture study showed a decreased capability of A. baumannii to adhere to the human epithelial cells at sub-inhibitory doses of colistin; however, none of the isolates could invade HEp-2 cells. Our study showed that the genes encoding biofilm-associated proteins undergo downregulation in expression levels after exposure to sub-MICs of colistin in A. baumannii . Longitudinal in vivo studies are needed to fully understand the clinical aspects of pathogenicity mechanisms and evolutionary dynamics of drug resistance. IMPORTANCE: Since the toxicity of colistin is dose dependent, there is a focus on strategies that reduce the dose while maintaining the therapeutic effect of the drug. Our findings about sub-inhibitory doses of colistin provide a novel insight into the logical use of colistin to treat and control Acinetobacter baumannii -related infections in clinical practice.

Lysostaphin is a glycylglycine endopeptidase, secreted by Staphylococcus simulans, capable of specifically hydrolyzing pentaglycine crosslinks present in the peptidoglycan of the Staphylococcus aureus cell wall. In this paper, we describe the cloning and expression of the lysostaphin enzyme gene in Bacillus subtilis WB600 host using pWB980 expression system. Plasmid pACK1 of S. simulans was extracted using the alkaline lysis method. Lysostaphin gene was isolated by PCR and cloned into pTZ57R/T-Vector, then transformed into Escherichia coli DH5α. The amplified gene fragment and uncloned pWB980 vector were digested using PstI and XbaІ enzymes and purified. The restricted gene fragment was ligated into the pWB980 expression vector by the standard protocols, then the recombinant plasmid was transformed into B. subtilis WB600 using electroporation method. The recombinant protein was evaluated by the SDS-PAGE method and confirmed by western immunoblot. Analysis of the target protein showed a band corresponding to 27-kDa r-lysostaphin. Protein content was estimated 91 mg/L by Bradford assay. The recombinant lysostaphin represented 90% of its maximum activity at 40 °C and displayed good thermostability by keeping about 80% of its maximum activity at 45 °C. Heat residual activity assay of recombinant lysostaphin demonstrated that the enzyme stability was up to 40 °C and showed good stability at 40 °C for 16 h incubation.

Background & Objective: The resistance genes encoding aminoglycoside-modifying enzymes (AMEs) are now widely prevalent in different populations of Staphylococcus aureus. The study aimed to determine the frequency of AMEs-encoding genes in clinical isolates of S. aureus.Methods: A total of 105 S. aureus isolates were obtained from the different clinical samples; and then were identified by conventional biochemical tests. The antibiotic resistance patterns of the isolates were characterized by the agar disk diffusion method. The distribution of the AMEs and femA genes was determined by conventional and multiplex PCR.Results: The aminoglycoside resistance rates of kanamycin, tobramycin, gentamicin, amikacin, and netilmicin were 47.6%, 46.6%, 45.7%, 45.7%, and 26.6%, respectively. 16.1% and 1.9% of isolates were MDR and XDR phenotypes, respectively. 21.9% of S. aureus isolates harbored the femA gene and were determined as methicillin-resistant S. aureus (MRSA) clones. The aac(6')/aph(2'') was the most prevalent (47.8%) AME-encoding gene in aminoglycoside-resistant S. aureus, followed by ant(4')-Ia (30.4%) and aph(3')-IIIa (21.7%).Conclusion:Our study demonstrated that the coexistence of several AMEs and the spread of the resistance determinants like femA in S. aureus clinical isolates are alarming and may contribute to the broadening of aminoglycoside resistance spectra and limit treatment options for staphylococcal infections.

Mosquito-borne infectious diseases such as dengue, Zika, chikungunya, and malaria continue to pose significant public health challenges worldwide. Traditional control methods, including the use of insecticides and environmental management, have shown limited effectiveness due to insecticide resistance and logistical challenges. Wolbachia-based strategies, which leverage the bacterium's unique ability to manipulate mosquito reproduction and block pathogens, present a promising, albeit not universally applicable, method for reducing the transmission of mosquito-borne diseases. Field trials have demonstrated significant but variable efficacy. This systematic review synthesizes the latest findings on Wolbachia-based interventions, focusing on their mechanisms, efficacy, and potential for large-scale implementation. We conducted a comprehensive literature search across multiple databases, including PubMed, Scopus, and Web of Science, to identify relevant studies published between 1990 and 2025. Our review highlights the promising results of Wolbachia-based strategies in reducing mosquito populations and disease transmission, while also addressing the challenges and limitations of this approach. We conclude that Wolbachia-based strategies hold great potential for revolutionizing mosquito-borne disease control, but further research and careful consideration of ecological and ethical implications are necessary for their successful implementation.