One Health & Risk Management

Genomics, One Health, and the Future of Infectious Disease Preparedness titlul la cuvint inainte

Dániel CADAR — 2025-12-23

In recent years, the landscape of infectious disease surveillance and diagnostics has undergone a profound transformation, driven by rapid advances in molecular biology, sequencing technologies, and bioinformatics. Emerging and re-emerging pathogens, increasing global mobility, climate-driven ecological changes, and intensified human-animal-environment interactions continue to challenge public health systems worldwide. In this context, the ability to detect, characterize, and monitor pathogens with high resolution and speed has become a cornerstone of effective preparedness and response.

Metagenomic and targeted genomic approaches now offer unprecedented opportunities to move beyond pathogen detection alone toward a deeper understanding of transmission dynamics, pathogen evolution, and population-level risk. These technologies enable unbiased discoveries of known and novel agents, support real-time outbreak investigations, and provide critical insights into genomic diversity that inform diagnostics, therapeutics, and public health decision-making. Importantly, they also bridge traditionally separated domains-human health, veterinary medicine, wildlife surveillance, and environmental monitoring embodying the principles of the One Health approach.

At the same time, translating these powerful tools into routine public health practice remains a complex task. Challenges related to sensitivity, standardization, cost-effectiveness, data interpretation, and integration into existing surveillance frameworks must be addressed thoughtfully and collaboratively. Capacity building, interdisciplinary cooperation, and the sharing of methodological experience across regions and institutions are therefore essential to ensure that genomic technologies deliver their full societal benefit.

This volume brings together expert perspectives that reflect both the promise and the practical realities of modern pathogen genomics. By highlighting methodological advances, applied case studies, and future directions, it contributes to a more informed and resilient public health infrastructure. I hope that the contributions presented here will stimulate dialogue, foster collaboration, and support the continued integration of genomic approaches into evidence-based public health action.

IN VIVO evaluation of gold nanoparticles functionalized with Arthrospira platensis protein extract

Ludmila RUDI — 2025-12-19

Introduction. Gold nanoparticles (AuNPs) have gained increasing attention in nanomedicine due to their biocompatibility and unique physicochemical properties, which make them suitable for targeted therapies against cancer and other chronic diseases. Functionalizing AuNPs with bioactive compounds further enhances their biocompatibility and therapeutic potential. This study aimed to evaluate the effects of 10 nm citrate-stabilized AuNPs functionalized with a protein extract from Arthrospira platensis (AuNPs-APE) on biodistribution, as well as hematological and biochemical parameters, in Wistar rats.

Material and methods. FTIR spectroscopy confirmed AuNPs' functionalization. Wistar rats received oral doses of AuNPs or AuNPs-APE for 28 days, followed by a 28-day clearance period. Gold biodistribution was analyzed in organs collected at the end of the experiment. Hematological and biochemical profiles were assessed using standard methods.

Results. FTIR analysis confirmed stable AuNPs-protein interactions. Both AuNPs and AuNPs-APE exhibited notable renal accumulation, suggesting extraglomerular retention. The protein extract showed mild immunomodulatory effects, reducing oxidative stress and inflammation associated with nanoparticle exposure.

Conclusions. The AuNPs–APE complex shows promise as a renal-targeted nanotherapeutic system, demonstrating improved biocompatibility and reduced systemic stress compared with unmodified AuNPs. Further studies are required to elucidate the underlying mechanisms and to optimize the formulation and safety profile.

Keywords. Gold nanoparticles, Arthrospira platensis, protein extract, Wistar rats, biodistribution, hematological assessment, biochemical profile.

Evaluation of Thymus vulgaris aqueous extract as a natural antimicrobial agent against urinary tract infection pathogens

Hashim ABASS — 2025-12-19

Introduction. Urinary tract infections (UTIs) are often caused by bacteria such as Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) which can lead to major health problems. The purpose of this study was to to examine the antibacterial effects of extract of Thymus vulgaris aqueous against these uropathogens.

Materials and Methods. The aqueous extract of Thymus vulgaris was prepared via hot water decoction (yield: 3.6 g from 30 g dried plant material 12%). Bacterial strains were isolated from 25 clinical urine samples and identified using selective culture media using the Gram staining and biochemical characterization. Antibacterial activity was assessed by using the agar well diffusion method with extract concentrations (50, 100 and 200 mg/mL) of Thymus vulgaris extract.

Results. The results of this study showed dose dependent of inhibition zone with S. aureus showing greater susceptibility (ZOI: 10–27 mm) compared to E. coli (ZOI: 9–23 mm). Ciprofloxacin shown larger inhibition zone (28–30 mm).

Conclusions. These findings demonstrate promising antibacterial potential of T. vulgaris aqueous extract as a natural alternative to combat antibiotic-resistant UTIs. These results support previous investigation about T. vulgaris as an alternative or complementary treatment for bacterial infections.

Biological properties of Serratia liquefaciens 1/2024, isolated from chicken

Liliіa VYGOVSKA — 2025-12-19

Introduction. According to a study by the European Centre for Disease Prevention and Control (ECDC) for 2022-2023, Serratia spp. caused 54,406 cases of disease in 28 countries worldwide. Serratia liquefaciens is considered the second most frequently isolated organism from human clinical samples after Serratia marcescens. The aim of this study was to investigate the biological properties of S. liquefaciens isolated from chickens.

Materials and methods. The study investigated a Serratia liquefaciens strain isolated in 2024 from chickens kept in a vivarium with clinical manifestations of gastrointestinal tract disorders. The aim of the study was to examine the strain S. liquefaciens. When studying the biological properties of the isolate, morphological characteristics, growth typicality, biochemical properties, pathogenicity for laboratory animals, and antibiotic sensitivity were taken into account.

Results. The isolated microorganism formed colonies typical of the Serratin genus, and the cells were motile at 37°C. The enzymatic profile matched the known characteristics of S. liquefaciens. The strain produced a denser biofilm at 23°C, exhibited pathogenicity in white mice, and showed resistance to several classes of antimicrobial agents. Based on the sequencing of a 16S rRNA gene fragment, the strain was deposited in the NCBI GenBank database under accession number PQ308601.1.

Conclusions. The results obtained suggest that the S. liquefaciens 1/2024 may be used as a reference strain for the differential diagnosis of enterobacteria, assessment of antimicrobial activity, and application as a laboratory control strain in experimental studies.

Single Nucleotide Polymorphism test for rapid detection of SARS-CoV-2 linеages in the Republic of N. Macedonia”

Gala MATEVSKA — 2025-12-23

Introduction. The coronavirus pandemic represents one of the most significant medical crises in recent history; therefore, rapid virus detection has become a critical component of public health practice. As the virus has undergone continuous mutations, the emergence of new variants has resulted in altered transmission dynamics, changes in disease severity, and implications for diagnostic testing. Although genomic sequencing is the most effective method for mutation detection, it remains time-consuming and expensive process.

Aim. To present a testing algorithm and evaluate the use of single nucleotide polymorphism (SNP) melting curve PCR for the detection of SARS-CoV-2 lineages, which may inform modifications in public health control and preventive measures, as well as potential adjustments to PCR-based diagnostic tests.

Materials and methods. RNA extracted from 140 SARS-CoV-2 positive samples received in the National Reference Laboratory for Virology, at the Institute of Public Health - Skopje as part of the COVID-19 surveillance system where Ct value ≤ 25 were subjected to SNP testing.

Results. Analysis of 140 SARS-CoV-2–positive samples collected between January and September 2022 using SNP testing revealed a predominance of the BA.4/BA.5 Omicron sublineages, accounting for 55.7% of cases.

Conclusions. Targeted SNP assays enable rapid and accurate detection of mutations associated with specific SARS-CoV-2 Omicron sublineages, facilitating early identification of emerging variants. These results may subsequently be subjected to further investigation, ultimately contributing to an improved public health response.

Clinical, biochemical, and genetic distinctions in patients with mitochondrial involvement versus other genetic disorders

Doina SECU — 2025-12-20

Introduction. Mitochondrial diseases are clinically and genetically heterogeneous disorders characterized by respiratory chain dysfunction, often mimicking other multisystem genetic conditions, necessitating an integrated clinical, biochemical, and molecular diagnosis. This study aims to compare the clinical, biochemical, and genetic profiles of patients with mitochondrial involvement and those with other inherited genetic disorders.

Material and methods. We analyzed 81 patients with suspected mitochondrial disease (Nijmegen Mitochondrial Disease Score ≥3), categorized into Group 1 (mitochondrial involvement) and Group 2 (other inherited disorders). Clinical, biochemical, instrumental, and molecular evaluations were performed using qPCR-HRM and Sanger sequencing, with data analyzed through descriptive statistics and non-parametric tests.

Results. Group 1 showed significantly higher rates of severe neuromuscular impairment, skill regression, ocular abnormalities, elevated plasma lactate and alanine, and characteristic neuroimaging findings, including basal ganglia abnormalities and cerebral-cerebellar atrophy. Genetic analysis identified phenotype-associated mutations in 32 patients, primarily affecting Complex I and V subunits and mitochondrial RNA genes, often involving multiple respiratory chain sites. Group 2 comprised a range of genetically confirmed non-mitochondrial disorders, identified through targeted genomic testing.

Conclusions. This study highlights the crucial role of integrated clinical, biochemical, and genomic approaches, emphasizing the importance of comprehensive molecular testing and multidisciplinary evaluation in addressing the diagnostic complexities of overlapping genetic disorders.

Astaxanthin and cellular metabolites production in Haematococcus lacustris exposed to silver nanoparticles

Liliana CEPOI — 2025-12-27

Introduction Silver nanoparticles (AgNPs) can modulate microalgal metabolism in a dose-dependent and stage-specific manner. Haematococcus lacustris, a key astaxanthin-producing microalga, is highly sensitive to environmental stressors that regulate the transition from the green vegetative stage to the red aplanospore stage.

Materials and Methods The effects of 10 nm and 20 nm citrate-stabilized AgNPs (0.01–5 mg L-1) were assessed on biomass, cellular metabolites, photosynthetic pigments, lipids, and astaxanthin. Nanoparticles were introduced on day 3, with analyses performed at the end of the green (day 9) and red stages (day 16).

Results Low to moderate AgNPs concentrations (0.01–1 mg L⁻¹) increased biomass, proteins, carbohydrates, and lipids during the green stage, while 5 mg L⁻¹ inhibited growth and pigments and elevated MDA levels. In the red stage, all concentrations reduced final biomass; however, 10 nm AgNPs at 0.01–0.5 mg L⁻¹ boosted astaxanthin (by up to ~30%) and lipids (by up to ~96%). Higher doses, along with all 20 nm AgNP treatments, suppressed astaxanthin accumulation.

Conclusions H. lacustris exhibits a hormetic response to AgNPs: mild exposure stimulates key metabolites, while higher concentrations become inhibitory. Nanoparticle size, dose, and timing are crucial for precisely directing metabolic pathways and improving astaxanthin yield.

Metagenomics at the interface of diagnostics and surveillance: a near-term perspective

Dániel CADAR — 2025-12-23

The global landscape of infectious diseases is undergoing rapid and profound change. Increased human mobility, climate-driven shifts in vector ecology, intensified human-animal-environment interactions, and geopolitical instability have collectively amplified the risk of emerging and re-emerging infectious diseases. At the same time, public health systems are increasingly confronted with pathogens that are unexpected, genetically diverse, or poorly represented in existing diagnostic panels. These developments highlight the growing limitations of exclusively targeted diagnostic approaches and underscore the need for broader, more adaptive tools.

In this context, metagenomic sequencing has emerged as a transformative technology. By enabling unbiased detection and genomic characterization of pathogens directly from clinical, environmental, or animal samples, metagenomics offers capabilities that extend far beyond conventional diagnostics. While historically confined to research and outbreak investigations, metagenomics is now approaching a level of maturity that warrants serious consideration for integration into routine diagnostics and risk-oriented surveillance frameworks.

Metagenomic sequencing is no longer a distant prospect but a rapidly maturing technology with tangible relevance for routine diagnostics and high-risk disease surveillance. While targeted assays will continue to form the backbone of diagnostic testing, metagenomics offers unparalleled breadth and genomic resolution, enabling detection and characterization of pathogens that would otherwise remain undetectable.

The near future of diagnostics will be defined not by the replacement of existing methods, but by their intelligent integration. By combining targeted testing with selective metagenomic analysis, public health systems can enhance preparedness, improve outbreak response, and strengthen biosafety and biosecurity in an increasingly complex global health landscape.