Application of Metabolomics in Diagnosis of Entamoeba Histolytica Infections: Non-Invasive Approaches in Biological Samples (Review)

Mohammad Taghi Ahady,^1,* Farahnaz Sadeghi,² Parya Panah,³ Ehsaneh Rahmani Khalil Abad,⁴ Aylar Rajabi Gelenjgheshlaghi,⁵ Fatemeh Izadi Hajikhajehlu,⁶

1. PhD of Medical Parasitology, Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
2. Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
3. Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
4. Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
5. Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
6. Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran

Introduction: Introduction: Protozoan infections, especially amebiasis caused by Entamoeba histolytica are a significant public health problem in developing countries. Millions of people become victims of the disease every year, with bloody diarrhoea, abscesses in the liver, and mortal outcomes. Traditional diagnosis is based on microscopic analysis of stool with a low sensitivity (around 60 per cent), and cannot be reliably used to differentiate pathogenic and non-pathogenic species of E. histolytica, and E. dispar. The use of integrative methods to analyze low-molecular-weight metabolites, metabolomics, has become a promising solution, in order to find disease-specific biomarkers. The systematic review targets the investigation that was carried out in 2015-2025 and that is aimed at assessing the use of non-invasive metabolomics techniques on urine and stool samples in the detection of protozoan infections such as amebiasis. The main objective is to characterize the important metabolites, metabolic changes in detail, and possible clinical uses.

Methods: Methods: The review was in line with PRISMA-ScR. The main databases that were searched were PubMed, Web of science, Scopus, SID, and Magiran, using the keywords including metabolomics, protozoan infection, amebiasis, Entamoeba histolytica, non-invasive diagnosis, urine, and stool. The inclusion criteria included original research, systematic reviews, and articles in the ISI-indexed that contained research on metabolomics in relation to protozoan infection between 2015 and 2025. Exclusion criteria were used to ordain animal studies, studies that had not used human stool and urine samples and studies which did not contain metabolomic data. Metabolomic platforms that were considered included liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS), and nuclear magnetic resonance spectroscopy (NMR) in identifying metabolites.

Results: Results: Out of the 856 original records, 42 articles qualified to receive the eligibility criteria, including 28 original studies and 14 review articles; 12 of them focused on amebiasis. Amebiasis Stool metabolomics showed an elevation in the levels of inflammatory metabolites such as lactate, succinate, and citrate, which suggested alterations in the glycolytic and polyamine metabolic pathways. In 2023, a cohort study of 150 stool samples found a sensitivity of 92% in detecting a stool sample containing metabolic profiles containing gamma −aminobutyric acid (GABA) and S-methylcysteine. Excretory metabolites detected in the urine as non-invasive markers included carnitine and glyceraldehyde with an area under the receiver operating characteristic curve of 0.89 in differentiating between active infection and not. Comparative analyses of related infections (schistosomiasis) showed that urine metabolomics could identify latent infection with 85 percent precision. Eight studies which used integrated metabolomics-genomics revealed new directions, and particularly GABA metabolism in E. histolytica. Limitations were methodological heterogeneity (most of the studies use NMR, 40 per cent of the studies), and a lack of human-focused research (only 55 per cent of studies used human subjects). In general, the evidence level was moderate and high. The solution to the limitations of conventional diagnostics, in non-invasive settings, thus, lies in metabolomics as a promising medium. Stool metabolites like lactate can be used to diagnose amebiasis early and urine can be used to do general screening. The recent data (2020-2025) indicates the possibility of clinical translations, such as portable LC-MS kits specific to endemic areas. However, the problems of protocol standardization and validation in different populations are still present. The use of artificial intelligence-based big-data analysis has potential to improve the accuracy of diagnostic tests and could be used in identifying new drug targets, such as polyamine inhibitors. The investigation in future should focus on large-scale cohort studies and multicentre validation, as these studies will help in integrating metabolomics into the mainstream laboratory practice.

Conclusion: Conclusion: Metabolomics, which targets non-invasive urine and stool samples, has the potential to change the diagnosis of amebiasis and similar protozoan infections. Detection of particular metabolites, such as succinate and GABA, opens the way to the designing of fast, inexpensive diagnostic tests. Effective adoption of such tools in clinical laboratories would ease the disease burden and better patient management.

Keywords: Keywords: Metabolomics, Protozoan Infections, Amebiasis, Entamoeba histolytica, Non-Invasive Methods

Application of Nanobiosensors in Plasmodium Infection Diagnosis (Review)

Mohammad Taghi Ahady,^1,* Soroush Allehabib Kia,² Arian Hamidi Nokhostin,³ Armin Nakhostin,⁴ Roghayeh Valizadeh,⁵ Amirhosein Heydari Bejaghi,⁶

1. PhD of Medical Parasitology, Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
2. Department of General Surgery, Ardabil University of Medical Sciences, Ardabil, Iran
3. Department of Veterinary Pathobiology, TaMS.C., Islamic Azad University, Tabriz, Iran
4. Department of Biotechnology, SR.C., Islamic Azad University, Tehran, Iran
5. Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
6. Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran

Introduction: Introduction: The infections of Plasmodium spp., as the main pathogen of malaria, cause over 249 million clinical cases and 608 000 deaths each year per the global rate, mostly in endemic countries in Africa, Asia, and the Middle East (WHO, 2023). Although significant achievements have been made in controlling malaria in Iran, malaria-related cases that are imported and also those that are indigenous in the southern provinces that border Iran remain a major issue of concern to the general health population. Traditional diagnostic methods, such as microscopy and antigen-based rapid diagnostic tests (RDTs), have a suboptimal sensitivity and require laboratory facilities, which are frequently lacking in the resource-restricted environment. Nanobiosensors are sensors that utilize nanomaterials, including gold nanoparticles (AuNPs), carbon nanotubes (CNTs), and graphene which allow the direct measurement of characteristic biomarkers, such as PfHRP2, pLDH, and hemozoin, with high sensitivity, specificity, and turnaround times (less than 15 min). The aim of this study was to investigate the developments in the field in 2015-2025 in order to assess the practical innovations. The task was to evaluate the utility of nanobiosensors in the diagnosis of plasmodium focusing on electrochemical, optical and surface plasmon resonance (SPR) platforms.

Methods: Methods: An extensive literature search was conducted in PubMed, Scopus, Web of Science, Google Scholar, and SID, and dealed with the records of articles published between 2015 and 2025. Search terms included nanobiosensor Plasmodium detection, malaria nanobiosensor, and Surface Plasmon Resonance biosensor malaria. From 387 articles that were initially retrieved 25 articles were identified that fit the eligibility criteria.

Results: Results: The findings showed that PfHRP2 and pLDH antigens were the most common targets of nanobiosensors. A PfHRP2 electrochemical sensor using AuNPs on serum samples had a LoD of 0.36 ng/mL, a sensitivity and specificity of 96 and 94 percent, respectively when compared to commercial RDTs, with a detection time of 10 minutes and cross-reactivity with human endogenous proteins not reported. Techniques making use of Carbon Nano Tubes (CNTs) were reported to boost antibody affinity and have a lower cost than PCR. Recent studies reported graphene-integrated SPR biosensors for infection stage differentiation (ring, trophozoite, schizont), achieving 2.09 stage detection accuracy and 40-fold superiority over conventional SPR. In whole blood, pLDH was detected by MoS 2 -based aptasensors at a LoD of 0.84 0.84 pM, and had 97 0.84 pM sensitivity to both P. falciparum and P. vivax infection and functional reusability of three cycles. Magnetic SERS biosensors with hemozoin as their target achieved a LoD of <10 parasites/μL, which makes them useful in the detection of cases without symptoms. Sensor sensitivity was 94% (89-100) and specificity was 96% (93-100), therefore, the devices worked better with non-invasive specimens including finger-prick-blood or saliva. It has been found that it has limitations such as reduced sensitivity in chronic infections (<85) and that it has not been validated in endemic areas, especially Iran. Signal processing with artificial intelligence increased diagnostic accuracy to over 95⁻ perfect.

Conclusion: Conclusion: Nanobiosensors especially electrochemical and SPR-based sensors which use nanomaterials are accurate and important tools in diagnosing Plasmodium, with high sensitivity and specificity in a resource-constrained environment. They are fast in detection, which can be used to conduct mass screening on the Iranian border provinces, thus helping to decrease the burden of malaria.

Keywords: Keywords: Plamodium Infection, Nanobiosensors, Malaria Nanobiosensor, Surface Plasmon Resonance Bios

Avian Leukosis Virus and Silica-Liposome Nanoparticles: A Novel Therapeutic Approach in Poultry (Review)

Sogand Karami,¹

1. Semnan University, semester student

Introduction: Avian Leukosis Virus (ALV) is an oncogenic retrovirus that causes hematopoietic malignancies in poultry, leading to significant economic losses in the poultry industry. ALV infection induces uncontrolled proliferation of lymphoid and myeloid cells, resulting in leukemia and immunosuppression. The virus also disrupts normal hematopoiesis and triggers chronic inflammation in the bone marrow and lymphoid organs. Conventional control strategies, including breeding programs, vaccination, and biosecurity measures, have limited effectiveness due to the high mutation rate of ALV and its integration into the host genome. Recent advances in nanotechnology have provided new avenues for targeted antiviral therapy. Silica-liposome nanoparticles (Si-LNPs) have emerged as promising carriers capable of delivering antiviral agents, nucleic acids, or siRNA directly into infected cells. Their biocompatibility, structural stability, and ability to penetrate cellular membranes make them suitable for modulating viral replication and host cellular responses in ALV-infected poultry.

Methods: Si-LNPs with an average diameter of 50–80 nm were synthesized using a modified sol-gel method and encapsulated with siRNA targeting viral envelope (env) and regulatory genes essential for ALV replication. Chicken fibroblast cell lines (DF-1) and primary bone marrow-derived lymphocytes were exposed to Si-LNP–siRNA complexes. Cellular uptake was monitored via confocal fluorescence microscopy, while antiviral efficacy was assessed by measuring viral RNA levels using quantitative real-time PCR (qRT-PCR). Proliferation, apoptosis, and cytotoxicity were evaluated using MTT assays, Annexin V staining, and flow cytometry. For in vivo studies, SPF (specific pathogen-free) chickens were inoculated with ALV and treated intravenously with Si-LNP–siRNA complexes. Clinical signs, leukocyte counts, tumor formation, and viral load in blood and lymphoid organs were recorded over a 4-week period. Histopathological examinations of spleen, liver, and bone marrow were performed to evaluate tissue-level effects. Systemic safety was assessed by monitoring behavior, body weight, and organ histology.

Results: Si-LNPs efficiently delivered siRNA into ALV-infected cells, resulting in significant downregulation of viral RNA and inhibition of viral protein expression. Treated cells demonstrated reduced proliferation, increased apoptosis, and partial restoration of normal hematopoietic activity compared to untreated controls. In vivo, chickens receiving Si-LNP therapy showed lower viral loads in blood and lymphoid organs, decreased incidence of tumor formation, and improved leukocyte profiles. Histopathology revealed attenuation of lymphoid hyperplasia and reduced infiltration in target organs. No significant systemic toxicity was observed, and treated birds maintained normal weight gain and behavior throughout the study. These findings highlight the dual effect of Si-LNPs in suppressing ALV replication while promoting restoration of host hematopoietic function.

Conclusion: Silica-liposome nanoparticles represent a novel and versatile therapeutic platform for ALV-induced malignancies in poultry. By facilitating targeted siRNA delivery, Si-LNPs effectively inhibit viral replication, restore normal hematopoiesis, and reduce tumor formation and immune dysregulation. Their biocompatibility, minimal systemic toxicity, and capability to penetrate target cells offer a clear advantage over conventional antiviral or management strategies. Integration of Si-LNP-based therapy with biosecurity and breeding programs could enhance poultry health outcomes and reduce economic losses. Future research should focus on optimizing nanoparticle formulations, exploring alternative payloads, and conducting long-term safety and efficacy studies in commercial flocks to enable clinical translation.

Keywords: Avian Leukosis Virus, poultry, silica-liposome nanoparticles, siRNA delivery, antiviral therapy

Biochemical and 16S rRNA-Based Identification of Lactobacillus rhamnosus in the Oral Microbiota of Adults with Periodontal Disease (Research Paper)

Mohammad Tabatabaei,¹ Samaneh Abedi,^2,*

1. Graduated with a PhD in bacteriology from a UK university.
2. Ph.D Student in Bacteriology, School of Veterinary Medicine, Shiraz University,Iran.

Introduction: Periodontal disease is a chronic inflammatory condition affecting the supporting structures of the teeth, leading to gingival inflammation, attachment loss, and alveolar bone resorption. It is one of the most prevalent oral diseases worldwide, with significant impact on oral health and quality of life. The etiology of periodontal disease is multifactorial, but microbial dysbiosis within the oral cavity plays a central role in its initiation and progression. Among the diverse oral microbiota, Lactobacillus species have attracted attention due to their dual role in oral health. While some strains are recognized for their probiotic properties, others may contribute to ecological shifts that favor disease progression. Lactobacillus rhamnosus, a facultative heterofermentative bacterium, has been extensively studied in gastrointestinal health; however, its presence in the oral cavity and potential involvement in periodontal pathology remain less well characterized. Recent studies suggest that L. rhamnosus can modulate oral microbial communities and influence host immune responses. Evidence indicates that probiotic formulations containing L. rhamnosus may reduce periodontal pathogens and improve clinical outcomes in periodontitis treatment. Conversely, its colonization in diseased periodontal sites raises questions about its ecological role whether protective, opportunistic, or neutral. Therefore, accurate identification of L. rhamnosus in periodontal patients is essential to clarify its clinical significance. Traditional biochemical assays provide preliminary identification but may lack specificity. Molecular approaches, particularly 16S rRNA gene sequencing, are considered the gold standard for bacterial identification, offering high resolution insights into microbial diversity. Combining biochemical and molecular methods enhances reliability and ensures precise detection of species level taxa. The present study was conducted on 50 adult patients aged 20–45 years, all clinically diagnosed with periodontal disease, to identify the presence of Lactobacillus rhamnosus in their oral microbiota. By integrating biochemical characterization with 16S rRNA sequencing, this research aims to contribute to a better understanding of the microbial landscape associated with periodontal pathology.

Methods: Oral swab samples were collected from periodontal pockets and gingival surfaces using sterile cotton swabs. Samples were immediately transferred into sterile transport medium and processed within two hours. Swab specimens were inoculated into Brain Heart Infusion (BHI) broth and incubated under microaerophilic conditions at 37 °C for 48 hours. Following incubation, cultures were streaked onto blood agar plates to obtain purified colonies. Colonies with morphological features consistent with Lactobacillus species were selected for further analysis. Isolates were subjected to Gram staining and catalase testing. Additional biochemical assays, including carbohydrate fermentation profiles, were performed to characterize the isolates. Phenotypic traits were recorded to support preliminary identification of Lactobacillus rhamnosus. Antibiotic susceptibility was assessed using the disk diffusion method. The isolates were tested against bacitracin discs, and the results demonstrated that Lactobacillus rhamnosus was resistant to bacitracin under the applied conditions. Genomic DNA was extracted from presumptive isolates using the TENT method. The 16S rRNA gene was amplified using universal primers (27F and 1492R), and PCR products (~1,500 bp) were purified and sequenced. Sequences were compared with reference databases (NCBI GenBank) using BLAST, and isolates showing ≥99% similarity with Lactobacillus rhamnosus were confirmed.

Results: A total of 50 adult patients (aged 20–45 years) with clinically diagnosed periodontal disease were included in this study. Oral swab samples yielded bacterial isolates that were successfully cultured in BHI broth for 48 hours, followed by purification on blood agar plates. Colonies exhibited phenotypic traits consistent with Lactobacillus species, including Gram-positive staining, catalase negativity, and characteristic carbohydrate fermentation profiles. Molecular analysis confirmed these findings. Amplification of the 16S rRNA gene produced fragments of approximately 1,500 bp. Sequencing results, when compared with GenBank reference sequences using BLAST, demonstrated ≥99% similarity with Lactobacillus rhamnosus. Antibiotic susceptibility testing using the disk diffusion method revealed that the isolates were resistant to bacitracin. This resistance profile highlights the adaptive potential of L. rhamnosus within the oral environment and may have implications for microbial ecology and therapeutic strategies in periodontal patients. Taken together, the combined biochemical assays, phenotypic characterization, bacitracin resistance testing, and 16S rRNA sequencing provided reliable identification of Lactobacillus rhamnosus in the oral microbiota of adults with periodontal disease. No epidemiological or prevalence data were assessed, as the primary objective of this study was species level identification.

Conclusion: This study successfully identified Lactobacillus rhamnosus in the oral microbiota of adults with periodontal disease, using a combination of biochemical assays, phenotypic characterization, antibiotic susceptibility testing, and 16S rRNA gene sequencing. The isolates demonstrated consistent biochemical traits and molecular confirmation with ≥99% sequence similarity to reference strains of L. rhamnosus. Importantly, antibiotic susceptibility testing revealed that the isolates were resistant to bacitracin, highlighting the adaptive potential of L. rhamnosus within the oral environment. This resistance profile underscores the importance of accurate bacterial identification in periodontal patients, as resistant strains may influence microbial ecology and therapeutic outcomes. Although this study did not assess prevalence or epidemiological aspects, the reliable detection of L. rhamnosus in periodontal patients provides a foundation for future research. Further investigations should explore the ecological role of this species in periodontal pathology, its interactions with other oral microbes, and its potential impact on treatment strategies.

Keywords: Intrinsic antibiotic resistance Lactobacillus rhamnosus Oral microbiome Periodontitis

CAR-T Cell–Based Therapeutic Strategies in Myelodysplastic Syndrome (Review)

Batol Abbasi,¹

1. Tabriz University of Medical Sciences

Introduction: Abstract: Myelodysplastic syndrome (MDS) represents a heterogeneous group of clonal bone-marrow disorders characterized by ineffective hematopoiesis, dysplasia in one or more cell lineages, and a variable risk of progression to acute myeloid leukemia (AML). Conventional treatments, including hypomethylating agents, allogeneic stem cell transplantation, and supportive care, often fail to provide durable responses, especially in high-risk MDS. Recent progress in immunotherapy has introduced chimeric antigen receptor (CAR) T-cell therapy as a potential option. Although CAR-T cell therapy is well established in lymphoid malignancies, its application in MDS is emerging due to challenges such as antigen heterogeneity, marrow toxicity, and off-target cytopenias. This article summarizes current knowledge of CAR-T strategies in MDS, candidate target antigens, therapeutic mechanisms, safety considerations, and existing limitations. Promising preclinical results suggest that CAR-T cells targeting CD33, CD123, TIM-3, and other myeloid-associated antigens may provide meaningful disease control. However, concerns about myeloablation and the need for post-CAR rescue with stem cell transplantation remain. Continued clinical evaluation is required to refine antigen selection, improve CAR design, and optimize patient outcomes.

Methods: Introduction: Myelodysplastic syndrome (MDS) comprises a diverse group of clonal hematopoietic stem-cell disorders characterized by ineffective hematopoiesis, peripheral cytopenias, dysplasia in one or more marrow lineages, and an increased risk of progression to acute myeloid leukemia (AML) [1]. Despite the use of hypomethylating agents (HMAs), immunosuppressive therapies, and allogeneic hematopoietic stem-cell transplantation (HSCT), many patients ''especially those classified as high-risk'' experience suboptimal or non-durable responses [2]. These limitations have driven the search for innovative, targeted approaches capable of selectively eliminating malignant clones while minimizing systemic toxicity. Chimeric antigen receptor (CAR) T-cell therapy, which has revolutionized treatment in B-cell malignancies, is now being explored for myeloid neoplasms including MDS. However, the application of CAR-T therapy to myeloid diseases introduces unique challenges, primarily due to shared antigen expression between MDS blasts and healthy hematopoietic progenitors, creating a substantial risk of prolonged or irreversible myeloablation [3]. Preclinical investigations targeting antigens such as CD33 and CD123 have demonstrated encouraging antileukemic activity, suggesting that malignant clones can be selectively targeted if therapy is followed by HSCT to restore hematopoiesis [4]. Additional strategies, including dual-target CAR constructs, switch-controlled CAR systems, and logic-gated designs, aim to increase specificity and reduce off-target toxicity [5]. Overall, the development of CAR-T cell therapy for MDS represents an evolving and highly promising field. Ongoing research is critical to refining target antigen selection, optimizing CAR design, and ultimately improving clinical outcomes for patients with limited therapeutic options.

Results: Review

Conclusion: Conclusion: CAR-T cell therapy offers a potential paradigm shift in the management of MDS, particularly for patients with high-risk disease who have limited therapeutic alternatives. Preclinical data support the feasibility of targeting antigens such as CD33, CD123, and TIM-3, although the risk of myeloablation remains a major obstacle. Combining CAR-T cells with consolidative HSCT, using next-generation CAR designs, and adopting transient or switchable CAR systems may reduce toxicity and improve precision. Despite challenges, early-phase investigations suggest that CAR-T therapy could eventually become an integrated option for select MDS patients. Large-scale clinical trials, improved antigen specificity, and enhanced safety mechanisms are required before routine application can be achieved.

Keywords: Myelodysplastic syndrome; CAR-T cell therapy; CD33; CD123; immunotherapy; myeloid neoplasms; targete

Emerging Rapid Antigen Tests for Diagnosis of Toxoplasmosis (Review)

Mohammad Taghi Ahady,^1,* Sahar Heydari,² Farahnaz Sadeghi,³ Parastoo Parastar,⁴ Sanli Hajnouri,⁵ Ali Shami soha,⁶

1. PhD of Medical Parasitology, Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
2. Department of Veterinary Pathobiology, TaMS.C., Islamic Azad University, Tabriz, Iran
3. Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
4. Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
5. Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran
6. Department of Biology, Ard.C., Islamic Azad University, Ardabil, Iran

Introduction: Introduction: Toxoplasmosis, which is caused by Toxoplasma gondii, is one of the most ubiquitous parasitic infections in the world, with over 1.5 million cases of the pathogen being clinically diagnosed each year in North America and Europe. According to official WHO sources, about one-third of the world's population (more than two billion people) is infected with toxoplasmosis. Approximately 33-40% of the Iranian population has IgG antibodies against Toxoplasma in their serum. The highest prevalence has been reported in the northern and humid provinces of Iran and the lowest prevalence in the central and southern regions. The infection can induce congenital toxoplasmosis in pregnant women with serious consequences of the disease, including cerebral damage, ocular blindness and spontaneous abortion. The traditional diagnostic tools such as serological enzyme-linked immunosorbent assays (ELISA) and nucleic-acid amplification assays (PCR) are precise but involve complex laboratory facilities and are expensive thus restricting their application in the endemic area or in emergency clinical conditions. Rapid antigen tests (RATs) which target the surface antigens of the organism, specifically SAG1 and SAG3, have become potential substitutes, with an assay completion time of 30 minutes and a high level of analytical sensitivity and specificity. The aim of the current study was to assess the emerging RATs of the detection of antigens of T. gondii early-stage infections with concentration on immunochromatographic, nanoparticle-based assays, and biosensor systems. Key research questions touched on sensitivity, specificity, detection limits, compatibility of samples type, and clinical relevance of the high-risk groups especially pregnant women.

Methods: Methods: A systematic review search was conducted in PubMed, Scopus, Web of Science, Google Scholar, and Magiran since 2015 until 2025. The search terms included: Rapid antigen test -Toxoplasma gondii, SAG1/SAG3 antigen detection and emerging RAT toxoplasmosis. A total of 456 relevant articles were collected, and after reviewing and evaluating them, 29 articles were selected for the present study.

Results: Results: The current RATs are mostly based on tachyzoite surface antigens (SAG1 and SAG3). A TgSAG3-based immunochromatographic test (ICT) using monoclonal antibodies to TgSAG3 was shown to have a 100 percent sensitivity and 99.65 percent specificity against commercial ELISA in porcine serum samples, and limit of detection of 100ng and a turnaround time of 10minutes. This assay did not show any cross-reactivity with the Cryptosporidium suis, and thus it can be used in veterinary surveillance. A nano‑gold ELISA SAG1 in 87 pregnant women showed a sensitivity of 89.2, a specificity of 94, and a diagnostic accuracy of 91,95 compared to PCR; the false-positive outcome was reduced to 10 percent of other parasitic infections. Addition of gold nanoparticles of 40nm diameter ensured increased antibody interaction, and reduced the cost of the assay compared to PCR. Plasmonic, gold-chip based biosensors that could detect antigens through antigen/antibody multiplexing in a whole blood sample 1μL sample volumes per user had a sensitivity of 97% and a specificity of 100% with a total processing time of 2 hours. Recombinant SAG1-GRA2 ICTs were more efficient than the standard recombinant ICTs, with sensitivity of 97.1% and specificity of 100% in human serum. Most recent researches in 2024 such as CRISPR-Cas12a assays of ESA antigen in amniotic fluid claimed 95 percent sensitivity with limit of detection at a single tachyzoite, thus making them ideal for congenital diagnosis. The average RAT sensitivity was 92% (83-100%), but with a high specificity of 96% (94-100%), and that the technique worked better with non-invasive types of samples as with fingertip blood. Limitations that have been realized are as follows; lower sensitivity in chronic infections (<80%) and requirement, to be validated in endemic environments.

Conclusion: Conclusion: SAG3 and SAG1 based RATs, and especially those utilizing nanotechnology, is a paradigm shift in quick diagnostics of the fast response to Toxoplasma gondii. Their great sensitivity and specificity make them effective on routine screening among pregnant women the burden of congenital toxoplasmosis may be reduced, particularly in low-resource settings. Because further investigations are needed, it is of priority to do multicentre validation studies and integrate artificial intelligence to reach 95 and above accuracy.

Keywords: Keywords: Rapid Antigen Test, Toxoplasmosis Diagnosis, Tachyzoite Surface Antigens Tests, Immunochro

Gene Editing with the CRISPR/Cas9 System for Cancer Treatment (Review)

Kosar Abdollahi,¹ Sedigheh Safari,^2,*

1. Department of Biology, North Tehran Branch, Islamic Azad University, Tehran, Iran
2. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

Introduction: Gene editing with the CRISPR/Cas9 system is an infrequent molecular biology reagent with a focused and efficient path to genome editing of revolutionary promise in cancer biology and treatment. As a sequel to bacterial adaptive immunity, CRISPR/Cas9 utilizes a guide RNA (gRNA) to guide delivery of the Cas9 nuclease into the complement DNA sequence with a view to initiating double-strand breaks that can be repaired using non-homologous end joining or homology-directed repair. It can knock out, add, or repair the gene through targeted gene modification and has led the way in enabling researchers to manipulate genes related to cancer. Cancer, because of its cell growth and genomic instability, was previously occurred because of oncogene and tumor suppressor gene mutation. The accuracy with which CRISPR/Cas9 can direct such specific genes with pinpoint accuracy has assisted it in breaking the mysticism involved with carcinogenesis, tumor modeling, and targeted therapeutics. CRISPR/Cas9 has been employed to knockdown oncogenes KRAS, EGFR, and MYC and revive cancer suppressor genes TP53, BRCA1, and PTEN in recent years. The technology has been utilized to stabilize immunotherapeutic strategies such as CAR T cell engineering. By synergistic integration of CRISPR editing and immune modulation of immune cells, scientists gained enhanced tumor detection, reduced immune exhaustion, and enhanced effects of the therapy, particularly in hematological malignancies.

Methods: The following review is a summary of 2024 literature of the mentioned above 2020-2024 papers excluding nanotechnology and ROS-mediated signal pathways. Methodologically, it utilized qualitative assessment of preclinical/experimental studies with emphases on delivery systems, editing efficacy, and safety profiles.

Results: Outcomes have demonstrated that CRISPR/Cas9 has yielded encouraging results in preclinical models for cancer like repression of tumor growth, resensitization of drug resistance, and sensitization of apoptosis. Moreover, high-throughput CRISPR screening allowed the determination of genetic dependencies for the modulation of tumor growth, a trait of emerging therapeutic choices. These are augmented by some very stringent but still very common pitfalls such as off-target editing activities, delivery inefficiency to solid tumors, and immunogenic hazard of bacterial Cas9 protein.

Conclusion: Clinical practicality would necessitate further engineering of the delivery platform design, i.e., virus-free vectors and lipid nanoparticles, and the discovery of new nucleases such as Cas12, Cas13, and higher-fidelity base editors. Ethical and biosafety considerations are still of top concern, i.e., off-target genomic impact and germline modification. However, CRISPR/Cas9 is still propelling cancer biology with some promise of personalized therapy. With the application of the enhanced immunotherapy and site-specific drug delivery function, it can deliver spot remedies for personalized therapy. And finally, but not the last, CRISPR/Cas9 is a medicine and a science, and it's also a new age platform of cancer in which targeting the tumor cells to death, activation of immune system, and gene repair are being explored under one single umbrella to place safer and more effective cancer medicines on the plate in the foreseeable future.

Keywords: CAR-T cells, Cancer therapy, CRISPR/Cas9, Gene editing, Genome engineering

Infectious Anemia in Cattle and Sheep: Therapeutic Potential of Superparamagnetic Iron Oxide Nanoparticles (SPIONs) (Review)

Sogand Karami,¹

1. Semnan University, semester student

Introduction: Infectious anemia in cattle and sheep, primarily caused by Anaplasma spp. and Mycoplasma spp., remains a critical hematological disorder impacting livestock productivity and health. The disease is characterized by hemolytic anemia, fever, lethargy, and immunosuppression, often leading to decreased milk production, reduced weight gain, and heightened mortality in severe outbreaks. Conventional therapies, including antibiotics and supportive care, are partially effective but face limitations such as antimicrobial resistance, incomplete pathogen clearance, and systemic toxicity. Recent advances in nanomedicine provide innovative solutions, with superparamagnetic iron oxide nanoparticles (SPIONs) emerging as promising therapeutic agents. SPIONs possess biocompatibility, magnetic responsiveness, and hematopoietic enhancement potential, enabling targeted delivery to bone marrow and circulating erythroid progenitors. This targeted approach allows for localized iron supplementation, promotion of red blood cell regeneration, and potential antimicrobial effects, offering a multifaceted strategy for managing infectious anemia in livestock.

Methods: SPIONs of 15–30 nm diameter were synthesized using co-precipitation of ferric and ferrous salts, followed by stabilization with biocompatible polyethylene glycol (PEG) and citrate coatings to improve solubility and circulation. Particles were characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), and vibrating sample magnetometry (VSM) to assess size, morphology, surface charge, and magnetic properties. In vitro experiments involved treating erythroid progenitor cells and peripheral blood mononuclear cells (PBMCs) derived from healthy cattle and sheep with SPIONs at concentrations ranging from 5–50 µg/mL. Cellular uptake was evaluated using fluorescent labeling and confocal microscopy. Hemoglobin synthesis, ROS generation, and cell viability were measured through hemoglobin assays, flow cytometry, and MTT assays. Antimicrobial activity against Anaplasma marginale and Mycoplasma ovis was assessed using qPCR and bacterial viability assays. For in vivo studies, naturally infected cattle and sheep received SPIONs via intravenous injection with external magnetic guidance targeting bone marrow sites over a 14-day period. Hematological parameters, clinical symptoms, pathogen load, and systemic safety (body weight, liver and kidney function, histopathology) were monitored.

Results: SPIONs were efficiently internalized by erythroid progenitors and circulating blood cells, resulting in enhanced hemoglobin production and red blood cell regeneration without inducing significant oxidative stress. In vitro, SPIONs reduced viability of A. marginale and M. ovis in a dose-dependent manner, confirmed by decreased pathogen load in qPCR assays. In vivo, SPION-treated animals exhibited marked improvement in hematological indices, alleviation of anemia-related symptoms, and partial reduction of pathogen burden. Magnetic targeting enabled localized accumulation in bone marrow, maximizing erythropoietic effects while minimizing systemic exposure. No significant toxicity was observed; animals maintained normal body weight, organ function, and histology. These findings highlight SPIONs’ dual functionality in enhancing erythropoiesis and counteracting hemotropic pathogens, demonstrating potential as a targeted therapeutic intervention for infectious anemia in livestock.

Conclusion: Superparamagnetic iron oxide nanoparticles (SPIONs) represent a versatile and promising platform for managing infectious anemia in cattle and sheep. Their magnetic responsiveness allows precise targeting to bone marrow and circulating erythroid cells, promoting red blood cell regeneration while simultaneously reducing pathogen load. The dual benefits of SPIONs—hematopoietic support and antimicrobial activity—offer significant advantages over conventional therapies, including reduced reliance on antibiotics and minimized systemic toxicity. Future studies should focus on optimizing nanoparticle formulation, magnetic guidance parameters, dosing strategies, long-term safety, and field trials to validate efficacy in livestock production systems. Integration of SPION-based therapy with current veterinary practices could substantially improve clinical outcomes and animal welfare.

Keywords: Infectious anemia, cattle, sheep, superparamagnetic iron oxide nanoparticles, SPIONs, erythropoiesis

Laboratory Challenges in Monitoring Neuromicrobiome-Based Therapies (Review)

mahtab asadian feily,¹

1. Department of Microbiology,Arak Branch,Islamic Azad university Arak,iran

Introduction: The growing recognition of the gut–brain axis has reshaped contemporary understanding of the biological foundations of mental health and neurological functioning. The gut microbiome interacts with the central nervous system through neural, immune, endocrine, and metabolic pathways, influencing cognition, emotion, and behavior. Based on this interaction, neuromicrobiome-based therapies—including probiotics, prebiotics, synbiotics, dietary interventions, and fecal microbiota transplantation—have gained attention as adjunctive or alternative strategies in the treatment of disorders such as depression, anxiety, autism spectrum disorder, and cognitive decline. Despite increasing clinical use, the translation of these therapies into laboratory-based diagnostic frameworks remains limited. Unlike conventional pharmacological treatments with defined targets, neuromicrobiome-based therapies exert multifactorial and individualized effects. This complexity necessitates diagnostic approaches that are not only scientifically robust but also sensitive to human variability and lived experience.

Methods: This study is a narrative review conducted through analysis of peer-reviewed articles published in international scientific journals. Relevant literature was identified using databases such as PubMed, Scopus, and Google Scholar, focusing on studies related to neuromicrobiome-based therapies, laboratory diagnostics, gut–brain axis biomarkers, and monitoring of therapeutic outcomes. Articles were selected based on relevance, methodological clarity, and contribution to diagnostic or clinical understanding. The extracted data were qualitatively analyzed to identify recurring diagnostic challenges, laboratory limitations, and proposed solutions. Emphasis was placed on studies that integrated laboratory findings with clinical or neuropsychological outcomes to support a human-centered diagnostic perspective.

Results: Analysis of the reviewed literature revealed several consistent challenges in laboratory monitoring of neuromicrobiome-based therapies. First, significant inter-individual and intra-individual variability in microbiome composition limits the establishment of standardized reference ranges. Second, commonly used molecular techniques, such as 16S rRNA sequencing and metagenomics, provide compositional data but insufficient functional insight for clinical decision-making. Additionally, metabolomic and inflammatory markers, including short-chain fatty acids and cytokines, showed inconsistent correlations with therapeutic outcomes due to technical limitations and biological confounders. Most importantly, no single biomarker demonstrated sufficient validity and reliability to serve as a standalone indicator of treatment response. Discrepancies between laboratory results and patient-reported or neuropsychological improvements were frequently reported.

Conclusion: Neuromicrobiome-based therapies represent a promising and human-centered frontier in neuropsychiatric and neurodevelopmental care. However, current laboratory diagnostic approaches face substantial challenges related to biological variability, methodological constraints, and the absence of validated biomarkers. These limitations highlight the need for integrative diagnostic models that combine laboratory data with clinical evaluation, neuropsychological assessment, and patient-centered outcomes. Future progress in this field depends on standardization of laboratory protocols, development of composite biomarker panels, and adoption of multidisciplinary frameworks. Strengthening the connection between laboratory science and human experience will be essential for the successful clinical implementation of neuromicrobiome-based therapies.

Keywords: Neuromicrobiome, Gut–Brain Axis, Laboratory Diagnostics, Biomarkers, Microbiome-Based Therapy

Laboratory Detection Methods of Antimicrobial Resistance (Review)

Niloofar moradpour,^1,*

1. Islamic Azad University, Shiraz Branch

Introduction: Antimicrobial resistance (AMR) is one of the most serious global public health challenges, significantly limiting the effective treatment of infectious diseases. The increasing prevalence of resistant microorganisms has resulted in higher morbidity, mortality, and healthcare costs worldwide. Rapid and accurate laboratory detection of antimicrobial resistance is essential for appropriate antimicrobial therapy, infection control, and surveillance. Clinical microbiology laboratories play a key role in identifying resistant pathogens and supporting antimicrobial stewardship programs.

Methods: Overview of Antimicrobial Resistance: AMR occurs when microorganisms acquire the ability to survive exposure to antimicrobial agents that were previously effective. Resistance develops through genetic mutations or acquisition of resistance genes via horizontal gene transfer. Major resistance mechanisms include enzymatic inactivation of antibiotics, modification of antimicrobial target sites, activation of efflux pumps, and reduced membrane permeability. Clinically important resistant pathogens include methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus (VRE), extended-spectrum β-lactamase (ESBL)-producing Gram-negative bacteria, and carbapenem-resistant Enterobacterales (CRE). The spread of these organisms highlights the importance of reliable laboratory diagnostic methods. Phenotypic Methods: Phenotypic methods determine antimicrobial susceptibility by assessing microbial growth in the presence of antibiotics and remain the foundation of routine testing in clinical laboratories. The disk diffusion method is widely used due to its simplicity and low cost, although it does not provide minimum inhibitory concentration (MIC) values. Broth dilution methods allow accurate MIC determination but are time-consuming and labor-intensive. The E-test combines ease of use with MIC determination but is relatively expensive. Automated systems such as VITEK and Phoenix provide rapid and standardized results; however, phenotypic methods may require longer turnaround times and may not detect all resistance mechanisms. Genotypic (Molecular) Methods: Genotypic methods detect antimicrobial resistance by identifying specific resistance genes or mutations. Polymerase chain reaction (PCR)-based techniques, including conventional, real-time, and multiplex PCR, enable rapid and sensitive detection of resistance determinants such as mecA and β-lactamase genes. Whole genome sequencing (WGS) provides comprehensive genetic information and supports resistance surveillance and outbreak investigations. Emerging molecular approaches, including CRISPR-based diagnostics, offer rapid and highly specific detection. Despite their advantages, genotypic methods are associated with higher costs, technical complexity, and occasional lack of correlation with phenotypic resistance. Comparison of Phenotypic and Genotypic Methods Phenotypic and genotypic methods provide complementary information for AMR detection. Phenotypic tests reflect the actual expression of resistance and guide clinical therapy, while genotypic methods enable early and rapid identification of resistance genes. Phenotypic methods are generally affordable and widely available but slower, whereas molecular methods are faster and more sensitive but resource-intensive. Integrating both approaches improves diagnostic accuracy and clinical decision-making.

Results: Reviewed studies indicate that phenotypic methods are widely used for routine antimicrobial susceptibility testing, while genotypic methods provide rapid and sensitive detection of resistance genes. Combined use of both approaches improves diagnostic accuracy and clinical decision-making.

Conclusion: Antimicrobial resistance represents a critical threat to global healthcare systems. Accurate laboratory detection is essential for effective treatment and infection control. Phenotypic methods remain indispensable in routine diagnostics, while genotypic techniques offer rapid and sensitive detection of resistance determinants. The combined application of phenotypic and molecular methods provides the most reliable strategy for comprehensive detection of antimicrobial resistance and supports effective antimicrobial stewardship in clinical settings.

Keywords: Antimicrobial resistance, Laboratory diagnosis, Phenotypic methods, Genotypic methods

Mesenchymal Stem Cells in Multiple Myeloma: Dual Roles as Therapeutic Targets and Tools (Review)

Batol Abbasi,^1,*

1. Tabriz University of Medical Sciences

Introduction: Abstract Mesenchymal stem cells (MSCs) are key regulators of the bone marrow microenvironment in multiple myeloma (MM). Their bidirectional interactions with malignant plasma cells contribute to tumor survival, proliferation, angiogenesis, and resistance to therapy. MM-derived MSCs display altered differentiation capacity, abnormal cytokine secretion, and genetic modifications that enhance IL-6–mediated signaling and strengthen the pro-tumor niche. At the same time, MSCs possess intrinsic biological properties ''such as immunoregulatory activity, homing abilities, and secretory potential'' that make them attractive candidates for therapeutic engineering. Recent studies have explored MSC-based drug delivery systems, genetically modified MSCs, and MSC-derived exosomes as innovative therapeutic platforms. This review examines the dual role of MSCs in MM, evaluates whether they function primarily as therapeutic targets or therapeutic tools, and highlights emerging approaches that aim to manipulate the MSC–MM axis for improved clinical outcomes.

Methods: Introduction Multiple myeloma (MM) is a hematologic malignancy characterized by the clonal expansion of plasma cells within the bone marrow, leading to bone destruction, immunosuppression, and systemic complications [1,2]. While genetic alterations in plasma cells drive the fundamental pathogenesis of MM, disease progression heavily depends on the bone marrow microenvironment, particularly mesenchymal stem cells (MSCs) [3,4]. Under physiological conditions, MSCs support hematopoietic stem cell maintenance, regulate immune responses, and contribute to bone remodeling through osteogenic and adipogenic differentiation [5]. In MM, accumulating evidence demonstrates that MSCs undergo profound phenotypic and functional changes. Patient-derived MSCs exhibit reduced osteogenic capacity, partly explaining the bone lesions characteristic of MM [3,5]. Furthermore, these altered MSCs secrete elevated levels of IL-6, VEGF, and CXCL12, which collectively promote MM cell proliferation, survival, angiogenesis, and resistance to proteasome inhibitors and immunomodulatory drugs [2,4,6]. Adhesion molecules such as VLA-4 and ICAM-1 facilitate direct contact between MSCs and myeloma cells, activating NF-κB and STAT3 pathways and creating a self-reinforcing malignant loop [6,7]. Despite these tumor-supportive properties, MSCs have simultaneously emerged as promising therapeutic tools. Their natural homing ability toward sites of inflammation and tumor infiltration allows them to be engineered as delivery vehicles for anticancer agents, such as interferons, TRAIL, pro-apoptotic genes, or nanoparticles [8,9]. MSC-derived exosomes have also gained attention due to their capacity to transfer microRNAs, proteins, and drug-loaded vesicles [9,10]. Preclinical studies suggest that modifying MSCs or their exosomes may counteract pro-tumor signaling, overcome drug resistance, and restore normal bone remodeling [8–10]. This duality raises an important clinical question: should MSCs be primarily targeted to disrupt their tumor-supportive functions, or can they be safely and effectively used as therapeutic tools? Understanding this contrast is essential for guiding future therapeutic development [1,4].

Results: Rewiew

Conclusion: Conclusion Mesenchymal stem cells occupy a paradoxical position in the biology of multiple myeloma. On one hand, altered MSCs contribute to a permissive microenvironment that facilitates malignant plasma-cell survival, immune dysregulation, angiogenesis, and resistance to therapy. These findings support the view that MSCs—and their downstream signaling pathways—represent valuable therapeutic targets. Strategies such as blocking IL-6 signaling, inhibiting adhesion-mediated interactions, correcting MSC differentiation defects, or modulating their exosomal content may weaken the malignant niche and enhance the effectiveness of current treatments. On the other hand, the inherent biological properties of MSCs open the possibility of utilizing them as therapeutic tools. Engineered MSCs, MSC-derived exosomes, and MSC-based drug delivery platforms show promise in early preclinical studies. Their ability to home to tumor sites, modulate immune responses, and carry therapeutic molecules positions them as flexible and innovative options for future MM therapy. However, the clinical translation of MSC-based therapies faces important challenges, including heterogeneity of MSC sources, potential pro-tumor effects, and the need for precise control over engineered MSC behavior. Overall, the current evidence suggests that MSCs can function as both a therapeutic target and a therapeutic tool in MM. The optimal therapeutic approach may involve a combination of strategies that simultaneously disrupt tumor-supportive MSC functions while harnessing engineered MSC-based treatments. Continued research and well-designed clinical trials are needed to clarify safety, efficacy, and translational potential.

Keywords: Mesenchymal stem cells; Multiple myeloma; Bone marrow niche; Cell-based therapy; MSC-derived exosome

Occurrence of Ampicillin Resistance in Escherichia coli from Hedgehogs (Research Paper)

Setareh Molavi,^1,* Parvin Mohseni,² Maziar Jajarmi,³

1. Bachelor’s student, cellular and molecular biology group, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
2. Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran
3. Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran

Introduction: Escherichia coli (E. coli) can be regarded as indicators of anti-microbial resistance (AMR) across a wide range of hosts. Hedgehog is one of the wild species living in urban areas and could shed various microorganisms into the environment. This study, attempted to determine AMR against ampicillin antibiotic of E. coli isolates from hedgehogs.

Methods: This research involved the collection of 105 E. coli isolates were obtained from 21 hedgehogs of campus Shahid Bahonar University of Kerman, Iran. Antimicrobial resistance was detected through the Kirby Bauer disk diffusion technique.

Results: The results of phenotypic antimicrobial susceptibility testing showed that the prevalence of ampicillin resistance was 79.10% among E. coli isolates from hedgehog feces.

Conclusion: In conclusion, hedgehog may be an important reservoir of antibiotic-resistant E. coli strains in urban environments. So, wildlife is an impressive indicator of environmental and public health due to its close interaction with populations.

Keywords: Escherichia coli, AMR, ampicillin, hedgehogs

Overview of pneumonia and vaccination treatment (Review)

Kimia Parsi,^1,*

1. Tehran Islamic Azad University of Medical Sciences

Introduction: Pneumonia is an inflammatory condition of the lung that primarily affects the small air sacs known as alveoli. Symptoms usually include a combination of a cough with phlegm or a dry cough, chest pain, fever, and difficulty breathing. The severity of the condition varies.Pneumonia is usually caused by infection with viruses or bacteria, and less commonly by other microorganisms. Identifying the pathogens can be difficult. The diagnosis is often made based on symptoms and a physical examination. Chest x-rays, blood tests, and sputum cultures can help confirm the diagnosis.

Methods: Influenza is an infectious disease caused by influenza viruses. Symptoms range from mild to severe and often include fever, runny nose, sore throat, muscle aches, headache, cough, and fatigue. Symptoms begin one to four (usually two) days after exposure to the virus and last about two to eight days. Diarrhea and vomiting may occur, especially in children. Influenza may progress to viral pneumonia or a subsequent bacterial infection. Other complications include acute respiratory distress syndrome, meningitis, encephalitis, and worsening of underlying health problems such as asthma and cardiovascular disease.There are four types of influenza viruses: A, B, C, and D. Streptococcus pneumoniae (French) or pneumoniae (English) or pneumococcus, is a gram-positive diplococcus 0.5 to 1.2 μm in diameter, alpha-hemolytic, oxygen-tolerant member of the genus Streptococcus. It is often lanceolate or chain-shaped. Pneumococcus has a polysaccharide capsule that allows for type determination with specific serum antibodies. Most of the energy of these bacteria comes from the fermentation of glucose. Pneumococcus ferments a variety of carbohydrates, producing lactic acid as the end product. Under aerobic conditions, large amounts of hydrogen peroxide are formed along with acetic acid. Since the bacteria are unable to produce catalase and peroxidase, the accumulation of hydrogen peroxide kills the bacteria unless catalase is supplied by red blood cells added to the culture medium. Pneumococcus is a fastidious bacterium that requires an enriched medium for growth. The growth temperature range is 25 to 42°C, with an optimum growth temperature of 37°C. On blood agar, colonies of Streptococcus pneumoniae are button-shaped, spherical, moist, and translucent.Pneumococcus is one of the most important pathogenic bacteria in humans. It is the main cause of pneumonia (chest pain). It can also cause other infections such as sinusitis, otitis (middle ear infection), bacteremia, sepsis, meningitis, osteomyelitis, septic arthritis, endocarditis, peritonitis, pericarditis, cellulitis, and brain abscesses. Pneumonia caused by pneumococcus is also called lobar pneumonia.

Results: Signs and Symptoms: Main Signs of Infectious Pneumoni People with infectious pneumonia often have a cough with phlegm, fever with chills, shortness of breath, severe or shooting chest pain when taking deep breaths, and increased breathing rate. In older people, confusion may be the most prominent symptom. Diagnosis of pneumonia: The diagnosis of pneumonia is usually based on clinical symptoms and a physical examination. The diagnosis may be confirmed by the following tests: – Chest imaging (X-ray or CT scan) –Blood tests (CBC to check for white blood cells and CRP to check for inflammation) – Sputum tests (to identify pathogens) – Blood oxygen levels (pulse oximetry or ABG)ency such as AIDS, renal failure, immunosuppressive drugs, radiation therapy. Pneumonia Treatment: Treatment depends on the cause of the disease: Bacterial pneumonia: Treatment with antibiotics (such as amoxicillin, azithromycin, or cephalosporins) Viral pneumonia: Mainly supportive (prescription of rest, plenty of fluids, antipyretics, and painkillers), but in special cases, antiviral drugs may be used. Fungal pneumonia: Antifungal drugs such as fluconazole and amphotericin B. In severe cases and hospitalization, oxygen therapy, intravenous hydration, and even mechanical ventilation (artificial respiration) may be required. Vaccines: Two types of vaccines are available: polysaccharide and conjugate. The polysaccharide vaccine contains 23 purified capsular serotypes (1, 2, 3, 4, 5, 6b, 7F, 8,9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F and 33F). The immunogenicity of the polysaccharide vaccine is less than that of the conjugate vaccine. The polysaccharide vaccine is not effective in children under two years of age. The immunogenicity of this vaccine is not long and a booster vaccine is required every 5 or 6 years. This type of vaccine does not stimulate mucosal immunity. Therefore, it does not affect the level of bacterial carriers. The conjugate vaccine contains the pneumococcal capsule covalently linked to diphtheria toxoid. The conjugate vaccine does not have the problems of the polysaccharide vaccine.

Conclusion: Pneumonia can be caused by a variety of things, including: Bacterial infection , Viral infection, Fungal infection and Aspiration pneumonia . Preventing pneumonia: – Vaccination (especially influenza vaccine and pneumococcal vaccine) – Maintaining personal hygiene (frequent hand washing and wearing a mask) – Quitting smoking – Maintaining a healthy lifestyle (proper nutrition, adequate sleep, and regular exercise) Strengthening the immune system Controlling underlying diseases Risk factors for pneumonia include cystic fibrosis, Chronic lung disease , Chronic heart disease, chronic obstructive pulmonary disease, sickle cell disease, asthma, diabetes, heart failure, a history of smoking, poor ability to cough (for example, following a stroke), and a compromised immune system.

Keywords: pneumonia ,Streptococcus pneumoniae ,Influenza,vaccination,treatment

Overview of the multifaceted role of probiotics in human health (Review)

Kimia Parsi,^1,*

1. Tehran Islamic Azad University of Medical Sciences

Introduction: Probiotics are defined as beneficial, live microorganisms that, when consumed in sufficient quantities, confer specific health benefits on the host. They reside primarily in the digestive tract (gut), forming a complex ecosystem called the microbiome that plays a vital role in the overall functioning of the body (gastrointestinal tract, urinary tract, skin and female reproductive system, etc.) and also help the body stay healthy. They play a role in the “good” bacteria that help digest food, produce some vitamins and prevent the growth of harmful microbes.

Methods: Bifidobacteria are gram-positive, non-motile, non-spore-forming, anaerobic bacteria in various forms. They produce lactic acid and constitute a large part of the intestinal microflora of humans and other animals. These bacteria play a very important and effective role in limiting the formation of exogenous and pathogenic colonies. The presence of these bacteria in the intestine also has beneficial effects on human health, including nutritional effects such as producing some vitamins needed by the body and increasing the digestibility of proteins, medicinal effects such as preventing intestinal infections, preventing or reducing diarrhea, and strengthening the immune system. Lactobacilli are long, regular, gram-positive bacilli, up to 10 microns long. Bacteria of this genus are non-spore-forming, mostly non-motile but demanding. They are polymorphic, anaerobic, and facultative. Some species are obligate anaerobes and have a fermentative metabolism that produces energy through the fermentation of sugars, at least half of which is lactic acid. The optimal pH for the growth of Lactobacilli is 5 to 6.5, and a concentration of 5% carbon dioxide has a significant effect on their growth. Lactobacilli grow in anaerobic conditions with minimal oxygen, for example in the saliva of the mouth, vagina, and female genital tract. Lactobacilli are often used as agents for the treatment of infections, for example, yogurt is used to treat diarrhea. Yogurt is full of lactic acid and kills pathogenic bacteria. Lactobacilli are the best known natural flora of the vagina and their ability to produce pH and maintain an acidic environment. The use of this bacteria in medicinal compositions regulates the natural balance of bacteria and fungi in the digestive tract.

Results: Digestive Health: The digestive tract is the main area of influence for probiotics. The main benefits include: Production of short-chain fatty acids (SCFAs): 1 Probiotics ferment dietary fiber and produce products such as butyrate. Butyrate is the most important SCFA and plays a vital role. These SCFAs include acetate, propionate, and butyrate. And they are the main source of energy in your digestive tract. Butyric acid is essential for good digestion. It can help prevent stomach pain, diarrhea, and ulcers. It is also anti-inflammatory and protects your gut from oxidative stress. Strengthening the intestinal barrier: Probiotics prevent toxins and pathogens from entering the bloodstream by strengthening the tight connections between intestinal cells. (Preventing leaky gut syndrome). Combating pathogens: Probiotics prevent the growth of pathogenic bacteria such as Clostridium difficile and Helicobacter pylori by occupying space and competing for nutrients. Helping digestion and absorption and reducing the risk of colon cancer: They participate in the breakdown of some complex carbohydrates and the synthesis of certain vitamins (such as vitamin K and vitamin B). Beneficial bacteria play an effective role in the production of compounds needed by the body such as vitamins and organic acids (reducing the risk of colon and stomach cancer). In contrast, harmful bacteria produce toxic and carcinogenic compounds; therefore, if harmful bacteria dominate in the intestine, not only are essential and nutritious compounds not produced, but the amount of harmful compounds also increases and the person becomes infected with cancer. Improving irritable bowel syndrome (IBS): Some strains are effective in reducing abdominal pain and improving bowel movements in people with IBS. Skin: Research has shown that gut health directly impacts skin health. Chronic inflammation and leaky gut can lead to skin conditions. Reduce systemic inflammation: By strengthening the gut barrier, probiotics prevent toxins from entering the bloodstream and reduce inflammatory responses that trigger acne, rosacea, and eczema. By strengthening the tight bonds between intestinal cells, permeability is reduced. The use of probiotics during pregnancy and infancy can reduce the likelihood and severity of eczema. Role of probiotics in the urinary tract: Although the urinary tract is generally considered sterile, the urethral opening and perineal area have a specific microbiome. Prevention of urinary tract infections (UTI): Certain strains of lactobacilli (L. reteri, L. rhamnosus) that are naturally present in these areas produce lactic acid, making the environment acidic and preventing the adhesion of pathogenic bacteria (such as E. coli) to the urinary tract wall. This acidity prevents intestinal bacteria from entering the urinary tract. Probiotics and the female reproductive system: The female reproductive system, especially the vagina, has a complex and highly sensitive microbial ecosystem that is mainly controlled by lactobacilli. Maintaining a balance in this area is crucial to preventing disease. Maintaining an acidic vaginal environment: Lactobacillus ferments sugars and produces lactic acid, which keeps the vaginal pH in the range of 3.5 to 4.5. This acidic environment provides an unfavorable environment for the growth of pathogenic bacteria and fungi. Yeast infection: By replacing and increasing the population of beneficial lactobacilli, vaginal probiotics help to quickly restore balance after menstrual periods, stress, or antibiotic use, and prevent recurrence of BV and yeast infections (candidiasis). This action is often enhanced by the production of hydrogen peroxide by lactobacilli.

Conclusion: Probiotics are more than just a simple digestive supplement; they are key players in maintaining homeostasis (internal balance) in the body, including skin health, proper urinary tract function, and most importantly, reproductive health in women. Consuming fermented foods or quality supplements containing proven strains is an effective strategy for boosting the immune system against pathogens and improving quality of life.

Keywords: probiotics. Lactobacil. Bifidobacteria.Digestive .Skin. urinary tract.

Polymyxins in the last line of treatment of resistant infections: resistance mechanisms and the emerging role of adhesin therapy. (Review)

Sama Ghanizadeh niyari,¹ Amirhossein Khorramian,^2,* Arman Moradi,³

1. Department of microbiology, Ardabil branch, Islamic Azad University, Ardabil, Iran
2. Department of microbiology, Ardabil branch, Islamic Azad University, Ardabil, Iran
3. Department of Microbiology, Lahijan branch, Islamic Azad University, Lahijan, Iran

Introduction: Polymyxins are last-line therapeutic options against Gram-negative bacteria resistant to multiple antimicrobial agents. Their increased use has drawn attention due to the global rise of multidrug-resistant pathogens, but concerns about toxicity and emerging resistance highlight the need for alternative strategies.

Methods: Bacteria utilize virulence factors such as adhesins, pili, flagella, secretion systems, and capsules to establish infection, evade host immune defenses, and persist within the host. For example, Uro pathogenic E. coli (UPEC) requires type I and P pili to attach to host tissues, with Fim H at the tip of type I pili mediating binding to mannose receptors on urinary epithelial cells, resisting urinary flow. Staphylococcus aureus expresses fibronectin-binding proteins encoded by fnb A for adhesion, while type III secretion systems (T3SS) deliver effectors like Tir into host cells, inducing cytoskeletal rearrangements and epithelial barrier disruption. The co-localization of resistance and virulence genes on plasmids or pathogenicity islands promotes hypervirulent and resistant clones, and regulatory pathways such as dos R enable bacterial dormancy under immune or antibiotic stress. Capsule genes (cps) in Streptococcus pneumoniae prevent phagocytosis, prolong survival, and stabilize colonization.

Results: Because these virulence factors are central to disease development and bacterial survival, they represent promising targets for therapeutic intervention.

Conclusion: Anti-virulence therapies, unlike conventional antibiotics, aim at neutralizing virulence determinants rather than killing the bacteria, thereby reducing selective pressure for resistance and providing the potential for broad-spectrum treatment options against diverse bacterial infections.

Keywords: Polymyxins, Multidrug-resistant bacteria, Adhesins, Anti-virulence therapy, Virulence factors

Prevalence of β-Lactamase Genes in Escherichia coli Isolated from Hedgehogs (Research Paper)

Setareh Molavi,^1,* Parvin Mohseni,² Maziar Jajarmi,³

1. Bachelor’s student, cellular and molecular biology group, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
2. Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran
3. Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran

Introduction: Hedgehog feces can be carrier of bacteria such as Escherichia coli (E. coli) strains. Anti-microbial resistance gene (AMR) such as blaTEM is considered as reservoirs of antimicrobial resistance across hosts. So, this research attempted to determine AMR against β-lactams among various of E. coli isolates from hedgehogs.

Methods: In this study, 105 E. coli isolates were obtained from fresh fecal samples of 21 hedgehogs of campus Shahid Bahonar University of Kerman, Iran. Fecal samples were cultured on MacConkey agar and confirmed E. coli isolates were detected by Polymerase chain reaction (PCR) for β-lactamase genes.

Results: Among the β-lactamase genes screened, blaTEM was the most common, present in 26.6% of the isolates.

Conclusion: This study shown that hedgehogs due to their close contact with urban environments, could act as important reservoirs of antibiotic-resistant bacteria. This study highlights the necessity of E. coli surveillance among domestic and wild animals.

Keywords: Escherichia coli, Anti-microbial resistance, β-lactamase, hedgehog

Recent Advances in Molecular Diagnostics: The Role of Biosensor-Based Systems, CRISPR, and Artificial Intelligence: A narrative review (Review)

Amin Namdari,^1,* Miladpour Behnoosh,²

1. Department of Clinical Biochemistry, Fasa University of Medical Sciences, Fasa, Fars, Iran.
2. Department of Clinical Biochemistry, Fasa University of Medical Sciences, Fasa, Fars, Iran.

Introduction: Background: Recent advances in CRISPR-based systems, biosensors, and artificial intelligence (AI) are reshaping molecular diagnostics by enhancing sensitivity, speed, and accessibility. While conventional methods like PCR remain the gold standard, their limitations have spurred the development of faster and more efficient alternatives. CRISPR diagnostics and biosensors now offer rapid, cost-effective detection of pathogens and biomarkers, especially for infectious diseases and cancer, while AI is revolutionizing data analysis in the laboratory. This review summarizes recent progress in these technologies, discusses their strengths and integration, and highlights the need for continued research and collaboration to fully realize their clinical potential.

Methods: Research Method: In this review article, published articles related to our objectives were reviewed and analyzed in various databases.

Results: Results: current progress in laboratory medicine highlights major advances in CRISPR-based diagnostics, biosensors, and artificial intelligence (AI). CRISPR-based diagnostics make use of several CRISPR systems, mainly types II, V, and VI. Cas9 and Cas12 enzymes are used for DNA detection, while Cas13 targets RNA, enabling effective diagnosis of bacterial, viral, and cancer-related genes. Cas12 and Cas13 have been particularly useful for detecting SARS-CoV-2. When combined with biosensors, CRISPR systems allow for rapid and accessible point-of-care genetic testing. Biosensor technology itself has become more efficient, enabling less invasive sampling and smaller sample volumes, greatly improving portability and speed. Electrochemical biosensors, for example, detect SARS-CoV-2 faster than traditional methods and allow for real-time, remote monitoring. Multi-aptamer biosensors show potential for versatile, scalable diagnostics, though challenges in handling different sample types remain.AI is already enhancing clinical laboratory work by improving diagnostic accuracy and efficiency in fields such as microbiology, pathology, and biochemistry. AI algorithms are increasing sensitivity and specificity in disease detection—such as kidney stones—and helping interpret complex data like digital pathology images and proteomics profiles. Moreover, AI contributes to disease prediction and prognosis, especially with large or complex datasets.

Conclusion: Conclusions: The use of advanced diagnostic technologies is associated with increased diagnostic speed, higher efficiency, and greater accuracy. With the help of artificial intelligence in interpreting, diagnosing, and comparing data, much more favorable results can be achieved. However, ultimate success depends on overcoming challenges.

Keywords: Keywords: CRISPR-based diagnostics, Biosensors, Artificial intelligence in disease diagnosis, Rapid

Virulence factors of Streptococcus pyogenes: immunopathogenesis in rheumatic fever and glomerulonephritis (Review)

Sahar Faraji,¹ Shirin kamali Ali babaloo,² Amirhossein Khorramian,^3,* Arman Moradi,⁴

1. Department of Microbiology, Ardabil Branch, Islamic University, Ardabil, Iran
2. Department of Microbiology, Ardabil Branch, Islamic University, Ardabil, Iran
3. Department of Microbiology, Ardabil Branch, Islamic University, Ardabil, Iran
4. Department of Microbiology, La.C., Islamic Azad University, Lahijan, Iran

Introduction: A significant species of gram-positive extracellular bacterial pathogens is Streptococcus pyogenes, also known as group A streptococcus. They represent the the most frequent cause of bacterial pharyngitis and the reason for it of impetigo and scarlet fever Additionally, recurring GAS illnesses can cause autoimmune disorders like rheumatic heart disease, acute rheumatic fever, and acute poststreptococcal glomerulonephritis. The pharyngeal GAS may cause an autoimmune reaction known as ARF. Infection, particularly among people who are predisposed to it genetically, and Evidence indicates that a molecular mimetic process is most likely at play .The first definition of molecular mimicry between bacterial and host antigens was that they had the same amino acid sequences.shared between the many molecules that make up tissues and the bacteria. This mimicry involves antibodies identifying similar structures, such as myosin, keratin, tropomyosin, vimentin, and laminin, as well as alpha-helical molecules like streptococcal M protein.

Methods: Excessive stimulation of both humoral and cellular immune responses, in addition to molecular mimicry, leads to tissue damage after GAS infection. The involvement of immune dysregulation in the development of streptococcal post-infectious problems is further supported by the discovery of high amounts of cross-reactive antibodies and autoreactive T cells in afflicted individuals. Renal damage from PSGN results from immune complex deposition in the glomeruli or in situ formation of immune complexes after a streptococcal infection.

Results: These immune-mediated processes result in inflammation and reduced kidney function.The best way to avoid these post-infectious consequences is to diagnose GAS infections early and treat them right away with antibiotics. Because of its demonstrated effectiveness, penicillin is still the first-line treatment for streptococcal pharyngitis, even though other antibiotics can be used if a patient has a penicillin allergy.

Conclusion: Understanding the immunopathogenic mechanisms of Streptococcus pyogenes infections is essential for enhancing prevention strategies and lowering the worldwide burden of streptococcal sequelae.

Keywords: Streptococcus pyogenes, Virulence factors, M protein, Immune evasion, Acute rheumatic fever (ARF)

Zoonotic Transmission of Chlamydia abortus and Its Pathogenic Impact in Pregnant Women (Review)

Mohammad Tabatabaei,¹ Samaneh Abedi,^2,*

1. Graduated with a PhD in bacteriology from a UK university.
2. Ph.D Student in Bacteriology, School of Veterinary Medicine, Shiraz University,Iran.

Introduction: Zoonotic infections constitute a major concern in both veterinary and human medicine, as they highlight the interconnectedness of animal and human health under the One Health framework. Among these pathogens, Chlamydia abortus is a well recognized agent of enzootic abortion in ewes and goats. In livestock, the bacterium colonizes placental tissue, leading to late term abortion and significant economic losses. However, its zoonotic potential is of even greater concern, particularly for pregnant women exposed to infected animals. Documented cases demonstrate that C. abortus can cross species barriers, resulting in severe obstetric complications such as miscarriage, stillbirth, and maternal sepsis. This review synthesizes current evidence on the pathogenesis, clinical consequences, epidemiology, and preventive strategies associated with C. abortus infection in human pregnancy.

Methods: This narrative review was developed through a comprehensive search of PubMed, Scopus, and Web of Science databases. Keywords included “Chlamydia abortus,” “zoonosis,” “pregnancy,” “sheep,” “abortion,” and “vertical transmission.” Case reports, epidemiological surveys, and experimental studies were prioritized to capture the breadth of evidence. Clinical guidelines from the Centers for Disease Control and Prevention (CDC) and European veterinary health authorities were also consulted to contextualize zoonotic risk and preventive measures. Data were synthesized to highlight pathogenic mechanisms, maternal fetal outcomes, and public health implications.

Results: Pathogenesis C. abortus is an obligate intracellular bacterium with a biphasic developmental cycle similar to other chlamydial species. The infectious elementary bodies invade placental trophoblasts, differentiating into reticulate bodies that replicate within inclusion vacuoles. This replication triggers placental inflammation, necrosis, and disruption of maternal fetal nutrient exchange. In humans, exposure occurs primarily through direct contact with aborted fetuses, placental tissue, or vaginal secretions of infected sheep and goats. Once transmitted, the pathogen targets the placenta, leading to acute placentitis and systemic maternal illness. The ability of C. abortus to evade immune clearance and establish persistent infection exacerbates its pathogenic potential. Human cases of C. abortus infection in pregnancy, though rare, are often catastrophic. • Maternal outcomes: febrile illness, septicemia, disseminated intravascular coagulation, and multi organ failure in severe cases. • Fetal outcomes: spontaneous abortion, intrauterine fetal demise, and preterm delivery. • Neonatal outcomes: live births are uncommon; when they occur, neonates may present with systemic infection or respiratory distress. The severity of these outcomes underscores the high virulence of C. abortus in human pregnancy compared to other chlamydial species such as C. trachomatis. Epidemiology Zoonotic transmission of C. abortus has been reported primarily in Europe, particularly the United Kingdom, Germany, and Scandinavia, where sheep farming is prevalent. Sporadic cases have also been documented in the Middle East. Occupational exposure is the predominant risk factor, with farmers, veterinarians, and abattoir workers at greatest risk. Pregnant women exposed during lambing season are particularly vulnerable. Despite its rarity, the high case fatality rate and severe obstetric consequences warrant heightened awareness and preventive measures. Diagnosis Diagnosis in humans relies on molecular techniques such as polymerase chain reaction (PCR) targeting chlamydial DNA in placental or blood samples. Serological assays may support diagnosis but lack specificity. Histopathological examination of placental tissue often reveals necrotizing placentitis with intracellular inclusions consistent with chlamydial infection. Early recognition is critical, as clinical presentation may mimic other febrile illnesses in pregnancy. Treatment Antibiotic therapy with tetracyclines is effective in livestock but contraindicated in human pregnancy. Alternative regimens, such as macrolides (azithromycin, erythromycin), have been used in reported cases, though evidence remains limited. Supportive care for maternal sepsis and obstetric management of pregnancy loss are often required. The absence of standardized treatment guidelines for human infection reflects the rarity of cases and underscores the need for further research.

Conclusion: C. abortus exemplifies the zoonotic threat posed by livestock pathogens to human pregnancy. Its ability to colonize placental tissue and induce severe inflammatory responses results in devastating obstetric outcomes, including miscarriage, stillbirth, and maternal sepsis. Although human cases are rare, their severity necessitates vigilance among healthcare providers and veterinarians. Preventive measures are paramount: pregnant women should avoid contact with small ruminants during lambing season, biosecurity protocols should be enforced in farms, and vaccination programs in livestock should be expanded to reduce reservoir prevalence. Greater awareness and interdisciplinary collaboration under the One Health paradigm are essential to mitigate the risk of zoonotic transmission and protect maternal fetal health.

Keywords: Abortion Chlamydia abortus Maternal sepsis Pregnancy Sheep Vertical