Journal of Biotechnology Research Center

Background: Escherichia coli are a versatile Gram-negative bacterium that is part of the normal intestinal flora of humans and animals. While most strains of E. coli are harmless and play a vital role in the gut microbiome, certain pathogenic strains can cause severe infections. Among these, uropathogenic E. coli (UPEC) is a prominent cause of urinary tract infections (UTIs), particularly in women. It is responsible for approximately 75–95% of uncomplicated UTIs and 40–50% of complicated UTIs worldwide, making it a significant public health concern. UPEC has evolved a range of virulence factors that enable it to colonize the urinary tract, evade the host immune system, and cause disease. These virulence factors include adhesins, such as type 1 and P fimbriae, which enable UPEC to attach to uroepithelial cells, and toxins like hemolysin and cytotoxic necrotizing factor 1, which damage host tissues. It also has mechanisms to acquire essential nutrients, such as iron, in the nutrient-limited environment of the urinary tract. Despite advances in understanding the pathogenesis of UPEC, UTIs remain a common and recurrent issue, partly due to the ability of UPEC to form biofilms and persist in the urinary tract. As antibiotic resistance among UPEC strains increases, there is a growing need for new therapeutic strategies to manage and prevent UTIs. Conclusion: Understanding the pathogenic mechanisms of UPEC is crucial for developing effective interventions against UTIs. Continued research into its virulence factors and resistance mechanisms is essential to address the challenges posed by recurrent and antibiotic-resistant infections.

Background: Pseudomonas aeruginosa is an opportunistic bacterium that causes nosocomial infections in hospitals. It can develop high-level multidrug resistance (MDR). The MexA gene is a constitutive chromosomal gene in the Resistance Nodulation Division (RND) family of P. aeruginosa. Objectives: This study aimed to detect the MexA efflux pump resistance gene in P.aeruginosa isolates. Methodology: A total of 165 specimens were collected from patients aged 5-65 years who suffered from burns, wounds, otitis media, and UTIs in two hospitals in Iraq during the period from November 2023 to January 2024. An antibiotic susceptibility test was performed using the Kirby-Bauer disk diffusion method in line with CLSI-2023. Finally, PCR was used to investigate the presence of the MexA gene in (25) P. aeruginosa isolates, which were highly resistant to Beta-lactam and quinolone antibiotics (MDR). Results: Out of 165 specimens, only 116 (70.30%) showed similar morphological traits and biochemical testing for Pseudomonas aeruginosa. Burn infections showed the highest isolation ratio, followed by wound infections, otitis media, and urinary tract infections (34.54, 17.59, 11.51, and 6.66) % respectively. Antibiotic resistance profile of (25) P. aeruginosa were 100% resistance to Ceftriaxone, Cefotaxime, and Amoxicillin-Clavulanic acid, while only 20% were resistant to Meropenem and Imipenem. MexA gene was found in 96% of P.aeruginosa isolates. Conclusion: The extensive identification of the MexA gene in P.aerogenosa indicates resistance to antibiotics associated with the function of the efflux pump gene and regulatory proteins.

Background: Klebsiella pneumoniae is a Gram-negative bacterium known to cause a variety of infections, especially in healthcare settings. It has the ability to form biofilms, which are organized communities of bacteria adhered to surfaces and surrounded by a self-produced extracellular matrix. Biofilms are important because they contribute to persistence and resistance to infection. FimH is a gene involved in biofilm formation and stress response in K. pneumoniae. Probiotics, live microorganisms that provide health benefits to the host, may affect the regulation of biofilm-related genes in pathogenic K. pneumoniae. The aim of this study was to investigate whether probiotics could influence the expression of biofilm-related genes, such as FimH, in K. pneumoniae. Methodology: One hundred and sixty samples, including patients with various infections such as burns, urine, sputum, wounds, lung fluid, catheters, pus, vaginal swabs, and ear swabs, were collected from hospitals in Baghdad, Iraq, between September 1, 2023, and November 30, 2023. The samples were streaked directly onto MacConkey agar, nutrient agar, blood agar, and colorimetric agar, then incubated at 37°C for 24 h. Bacterial isolates were accurately identified using the Vitek-2 system. Antibiotic susceptibility was tested using the Kirby-Bauer technique as recommended by CLSI (2023). Thirty isolates were tested for biofilm using microtiter plates. The study included molecular characterization of the biofilm gene fimH, while the ability of Lactobacillus fermentum to inhibit biofilm production was also examined. Results: Results showed that only 80 of the 160 samples had the same morphological and biochemical characteristics as Klebsiella , whereas only 50 of the 80 isolates were identified as Klebsiella pneumoniae. Antibiotic susceptibility showed that K. pneumoniae isolates exhibited varying degrees of resistance to the antibiotics used. The highest percentage of antibiotic resistance was found for Ceftriaxone (76.6%). In addition, Amikacin and Tigecycline had the lowest resistance rates, at 0% each. Molecular testing of 30 multidrug-resistant strains was performed by detecting the fimH gene. Results showed that all 30 K. pneumoniae isolates had this gene. Strong biofilm was detected in one isolate. Real-time PCR was used to detect fimH gene expression before and after probiotic and probiotic-combined antibiotic treatments.
Conclusion: The majority of K. pneumoniae isolates demonstrated the ability to form biofilms. All isolates have the fimH gene. Additionally, Combinations of L. fermentum and antibiotics exhibited synergistic effects against the KP1 isolate of K. pneumoniae compared with using the probiotic or antibiotic alone.

Background: Antimicrobial resistance has emerged as a major global health challenge, prompting the search for novel antimicrobial agents. Natural products, including quince (Cydonia oblonga Miller), are rich in bioactive compounds with potential therapeutic properties. Objective: To evaluate the antibacterial activity of aqueous and ethanolic extracts of quince fruit against pathogenic bacteria isolated from patients with laryngitis. Methods: Swab samples were collected from patients with laryngitis and cultured on selective media to isolate Gram-positive and Gram-negative bacteria. Isolates were identified using the VITEK system. Quince fruit extracts (aqueous and ethanolic) were prepared at concentrations of 25, 50, and 75 mg/mL and tested for antibacterial activity using the agar well diffusion method.
Results: The predominant isolates included Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. The ethanolic extract demonstrated the highest antibacterial activity, particularly against S. aureus (12 mm inhibition zone), followed by E. coli (9 mm) and P. aeruginosa (8 mm). The aqueous extract exhibited comparatively lower activity but followed the same sensitivity pattern.Conclusion: Quince fruit extracts exhibit notable antibacterial activity, with ethanolic extracts showing superior efficacy. These findings suggest potential applications in antimicrobial therapy and food preservation.

Background: Ovarian cancer (OC) remains the deadliest of all gynecological tumors. Because there are no obvious clinical symptoms, OC is difficult to diagnose early, which makes it an aggressive disease, highlighting the vital need for reliable biomarkers. This research aimed to investigate the role of ATP-citrate lyase and glycosyltransferase levels in ovarian cancer patients. Methodology: 140 women with ovarian cancer and control women aged 35- 65 years were categorized into three groups upon enrollment in this case-control study: 40 women as controls, 50 as group I(untreated), and 50 as group II (treated) were collected from Al-Amal Hospital for Radiation and Nuclear Medicine and Oncology Teaching Hospital at Baghdad Medical City. Results: The results showed elevated levels of ACLY and GTs enzymes in group I patients compared with the mean value recorded in the control group (P-value <0.0001). Whereas, the result showed no significant difference between group II and control (P-value =0.2149). A receiver operating characteristic curve analysis showed high sensitivity, as indicated by the area under the curve. Conclusions: The current study concluded that ACLY and GTs could be used as tools to help diagnose and predict ovarian cancer in the future.

Background: Streptomyces comprises Gram-positive, filamentous, and spore-forming bacteria. Streptomyces fradiae has the capability to synthesize the antibiotics neomycin, tylosin, and fosfomycin. Aim of the study: Detection of the inhibitory effect of S. fradiae isolates against microbial pathogens (in vitro) and study its compatibility with biosynthesized zinc oxide nanoparticles. Methodology: Fifty soil samples were gathered between December 2023 and January 2024. Specimens were gathered from several locations inside the city of Baghdad. Following the dilution of the sample, it was plated onto ISP2 agar to isolate Streptomyces. Multiple biochemical assays were conducted on the isolates. Zinc oxide nanoparticles were produced using Eucalyptus extract and evaluated against harmful microorganisms. Results: A total of 33 Streptomyces isolates were obtained, then identified as Streptomyces fradiae , 12 isolates did not exhibit any antibacterial effects against pathogenic bacteria. On the other hand, 21 isolates (63.6%) demonstrated strong antibacterial activity. Of the total isolates, 48.5% (16) exhibited antibacterial activity against both Escherichia coli (E. coli) and Staphylococcus aureus(Staph. aureus). However, only 15% (5 isolates) demonstrated activity, particularly against Staph. aureus. The zone of inhibition of the nanoparticles against the tested strains was 9mm, while for the Eucalyptus extract was 2.5mm against Staph. aureus. The inhibition zones against E. coli, on the other hand, were 11mm for nanoparticles and 3mm for the Eucalyptus extract. The effective concentration was 12.5% for the plant’s extract and 20% for nanoparticles. Conclusion: The Streptomyces’s bioactive molecule exhibited antibacterial properties. It was more effective against Gram-positive bacteria (Staph. aureus) than Gram-negative bacteria (E. coli). This study presents an environmentally friendly method for producing zinc oxide nanoparticles (ZnO NPs) using a leaf extract from Eucalyptus globulus as a reducing agent. The ZnNPs have antibacterial efficacy against both E. coli and Staph. aureus. The antibiotic of Streptomyces was more effective than Zn NPs and Eucalyptus plants against pathogenic bacteria

Background: Acinetobacter baumannii is an important opportunistic pathogen associated with urinary tract infections and notable multidrug resistance. Biofilm formation further complicates treatment and contributes to persistence in clinical settings. Objective: This study aimed to evaluate the antibacterial and antibiofilm activities of 3-iodoindole against multidrug-resistant A. baumannii isolates recovered from patients with urinary tract infections in Baghdad, Iraq. Methodology: A total of 150 urinary isolates were collected from hospitals and laboratories in Medical City, Baghdad. Isolates were identified by conventional culture methods and the VITEK 2 Compact system. Antimicrobial susceptibility was assessed by the Kirby-Bauer method and VITEK 2 AST cards. Biofilm formation was quantified using a microtiter plate assay, and the antibacterial and antibiofilm activities of 3-iodoindole were evaluated at different concentrations. Results: Of the 150 isolates, 100 (66.7%) were confirmed as A. baumannii. High resistance rates were observed against several commonly used antibiotics, whereas colistin and tigecycline remained the most active agents. Biofilm assays showed that 79% of isolates were biofilm producers, including moderate and strong biofilm-forming phenotypes. Strong biofilm formers exhibited significantly higher OD600 values at 24 h than weak biofilm formers. 3-Iodoindole demonstrated marked antibacterial activity, achieving a >3-log reduction in viability at 250 µg/mL within 15 min and significantly reducing biofilm biomass in a dose-dependent manner. Conclusion: 3-Iodoindole exhibited potent antibacterial and antibiofilm effects against multidrug-resistant A. baumannii and may represent a promising candidate for further development as an alternative or adjunctive antimicrobial agent.

Background: Jatropha integerrema Jacq. is cultivated in Iraq and recorded as an exotic genus. The leaves and flower extracts have medicinal properties for treating conditions such as HIV, ringworm, tumors, ophthalmia, malaria, bronchitis, and asthma. Objective: Quantitative analysis of Jatropha leaf and flower extract, estimation of total phenolics and terpenoids, and molecular analysis. Methodology: This research conducted a quantitative comparison of bioactive chemical constituents between leaf and flower extracts, and a phylogenetic analysis of two plant organs of J. integerrema Jacq., leaves and flower petals, using nrDNA (ITS) and two plastid genes (matK and trnT-trnL). Results: Phenols, terpenoids, alkaloids, and steroids in different concentrations were present in leaves and flowers. The phenolic compounds had the highest values, 498.8 and 305.98 mg/g, whereas the steroids had the lowest concentrations, 2.00 and 1.25 in leaf and flower extract, respectively. The plant exhibited higher free radical scavenging activity compared with ascorbic acid. The antioxidant activity of leaves was higher than that of flower extracts in scavenging the free radicals (85.33% for leaves and 75.13% for flowers at 200 µg/mL). Conclusions: This study estimated that the various chemical concentrations were derived from the leaves and flowers of J. integerrema, which contains many phytochemical components with beneficial properties. So it has significant potential as a novel source of medication. The nrDNA and plastid genes are effective tools for comparing plant organs and improving agronomic traits related to plastid genes.

Background: Humans are constantly exposed to a variety of microbes, making them vulnerable to numerous diseases. Among these, biofilm-associated infections form the largest category of diseases. Serratiopeptidase, recently referred to as the "miracle enzyme," has been widely utilized in Europe and Asia as a therapy to combat biofilms. Objective: This study aimed to clone and express the stp gene and to test the effectiveness of the expressed Serratiopeptidase as an antimicrobial and antibiofilm agent. Methodology: The gene encoding Serratiopeptidase was isolated from a pathogenic isolate of Serratiamarcescens after amplification of the stp gene, followed by confirmation through sequencing. Bacterial transformation was performed using the cloning vector pGEM-3Zf (+) after amplification and ligation of the gene encoding the protein. Then, it was transformed into the expression host E. coli BL21 (DE3) bacteria. IPTG was used to induce the bacterial culture to produce the protein. The presence of the Serratiopeptidase protein was confirmed through SDS-PAGE. Results: The anti-biofilm activity was validated using a titer plate assay. The results demonstrated significant effectiveness as an anti-biofilm agent. However, the expressed protein showed no antimicrobial activity against bacterial and fungal species. Conclusion: Expressed protein has anti-biofilm formation, but has no activity against pathogenic bacteria and fungi.

Background: Heavy metal pollution is one of the major environmental threats affecting aquatic ecosystems and phytoplankton communities in rivers. Algae are important bioindicators of water quality and may contribute to the bioremediation of pollutants. Objective: This study aimed to assess heavy metal contamination in selected areas of the Tigris River and evaluate its impact on non-diatom phytoplankton diversity and distribution. Methodology: Water and algal samples were collected from four sites along the Tigris River during autumn 2024. Physicochemical parameters, nutrients, and heavy metals (lead, nickel, and chromium ) were measured using standard laboratory methods. Algal species were identified microscopically, and statistical analyses were performed. Results: The results showed elevated concentrations of lead, nickel, and chromium with significant variations among study sites. A total of 28 non-diatom phytoplankton species belonging to 15 genera were identified. Cyanobacteria represented the dominant group, while Euglenophyta showed the lowest occurrence. Significant correlations were observed between heavy metal concentrations and algal community composition. Some algal species demonstrated tolerance to high levels of heavy metals. Conclusion: Heavy metal pollution in the Tigris River affects the diversity and distribution of phytoplankton algae. Certain algal species exhibited resistance to heavy metal contamination, indicating their potential application in bioremediation processes