In the post-COVID-19 era, there has been a notable surge in the development of mRNA vaccines. These vaccines are not only targeting various pathogens beyond SARS-CoV-2 but also hold promise in treating cancer and genetic disorders. These type of vaccines are revolutionizing vaccinology through their inherent possibility for rapid pandemic response, high efficacy, minimal side effects, and cost-effectiveness. Achieving these benefits hinges on seamlessly integrating mRNA production steps, from plasmid DNA (pDNA) design and antigen cloning, in vitro transcription to lipid nanoparticle formulation. A critical initial step in mRNA vaccine production is pDNA cloning vector design. The vector should be carefully constructed considering a copy number of plasmid, vector backbone with a promoter, the origin of replication, multiple cloning sites, polyadenylation signal, and markers for selection. However, despite careful design, challenges like poly-A tail deletion may arise, prompting the exploration of stable large-size and low-copy vectors, as well as linear and bacteriophage vectors. Additionally, for large-scale production and regulatory compliance, vector systems must be scalable and well-documented. This overview aims to elaborate upon the intricacies in pDNA cloning vector design. The focus is on achieving accurate insert sequence, especially those encoding the complex antigens and gene expression, highlighting the critical role of pDNA design in ensuring vaccine effectiveness. Although commercial vectors and automated synthesis facilitate gene construction, challenges still exist. This emphasizes that a multifaceted approach, combining molecular biology techniques, computational tools, and collaboration with experts in microbiology, molecular biology, and vaccine development, is required for successful mRNA vaccine development.

Escherichia coli is by far the leading cause of community-acquired UTIs. Its emerging antimicrobial resistance, along with the existence multidrug-resistant (MDR) strains is one of the most difficult global healthcare challenges. In Europe, including Serbia, E. coli isolates obtained from patients with acute uncomplicated urinary tract infections (UTIs) have increasing antimicrobial resistance, particularly to ciprofloxacin and trimethoprim-sulfamethoxazole. Pivmecillinam, the oral prodrug of the penicillin derivative mecillinam, was introduced in national and international protocols around the globe for UTIs treatment in ambulatory settings after showing high efficiency for uncomplicated low UTIs. During its application in various European countries, mecillinam has shown a satisfactory clinical effect against uropathogenic E. coli. Thus, it is commonly used as the anibiotic of choice for the empirical management of UTIs, mostly due to its low resistance rates and low damaging effects on the gut microbiota as a pro-drug. Research showed that mecillinam exhibits high antimicrobial activity against MDR E. coli strains, such is the case in metallo-β-lactamase (MBL)-producing isolates (both NDM-1 and IMP types) and OXA-48-like-positive strains. Mecillinam is yet to be registered in Serbia, in order to be tested as a potential first-choice antibiotic for UTI caused by E. coli. Thus, we review the current literature data on mecillinam’s antibacterial effect in this particular bacteria.

The “silent pandemic” – a WHO-coined phrase that relates to the rise and spread of antimicrobial resistance is a phenomenon so striking that in spite of living in the most prosperous times, we are threatened to return to the preantibiotic era when even the simplest of interventions and infections were possibly deadly. One of the possible solutions could be the application of natural predators of bacteria – bacteriophage therapy. Discovered over a century ago, phages have been studied extensively and have become irreplaceable tools in laboratory work. The knowledge they helped to generate is still enlarging, and was responsible, at least in part, for creating the whole scientific discipline of Molecular biology. Their application on the other hand has not been as continuous and fruitful. On the contrary, the history of phage therapy is one of rise and fall, enthusiasm and acceptance as well as rebuttal and oblivion, scientific misconceptions and dogmatism, bad corporate investments, politics and ideologies. As today we are equipped with far more knowledge on the biology of phages, bacteria and resistance than early phage enthusiasts, it is justified to attempt phage application once again. Here we elaborate on the early years and very first attempts of phage therapy and the troubles rising from those days as they serve as excellent cornerstones for future endeavors in the field. In parallel, the current state-of-the-art of phage therapy in different parts of the world is also presented.

Viral infections such as Epstein-Barr virus (EBV) and Herpes simplex virus (HSV) represent significant complications in patients with haematological malignancies and increase the frequency of morbidity and mortality. These infections represent one of the main factors of unsuccessful treatment; therefore, regular monitoring of active viral infections is necessary. The work aimed to determine the frequency and dynamics of active EBV and HSV infection in patients with haematological malignancies, monitored at the Institute of Microbiology and Immunology, Faculty of Medicine University of Belgrade over two years. This study included 35 patients treated at Hematology Clinics of the University Clinical Center of Serbia. Patients were monitored during 2020-2021. Ten blood samples were taken from each patient during monitored period. The samples were taken in a time interval between 2 weeks and three months, depending on the arrival of patients at medical controls. The first sample is marked with the first measurement of blood drawn during 2020 for each patient. The second measurement was a second blood sample taken in the previously mentioned time interval, and so on up to the tenth sample marked as the tenth measurement. For the detection of EBV DNA, the “in-house” PCR method was used to detect the EBNA-1 gene. According to the manufacturer’s instructions, the commercial kit Ampli Sens® HSVI, II-FRT was used for HSV DNA detection (InterLabService, Moscow, Russia). The lowest frequency of EBV infection was during the second measurement (8.57%), while the highest was during the eighth and tenth measurements (31.43%). The highest frequency of HSV infection was during the eighth measurement (5.71%), while during the third, fifth and ninth measurements, none of the subjects had positive PCR for HSV DNA. A statistically significant difference was found in the frequency of EBV infection in later measurements compared to earlier measurements (p=0.002). In contrast, this was not the case in the study of HSV frequency infection (p=0.750). This study presents the dynamics of the frequency of active EBV and HSV infection in patients with haematological malignancies in two-year period. The frequency of active EBV infection was significantly higher compared to HSV infection.

Globalization has resulted in an increase in food demand, which requires advancements in farming techniques to meet the demands. Green revolution based agriculture has made our country self-sufficient in food production. However, the excessive use of agrochemicals has led to degradation of soil, water, environment and human health besides, reduction in biodiversity. This necessitates an alternative of agro-chemicals which are environmentally safe and free from pesticide residues. Under such situation, Natural Farming as promoted by Padam Shri Awardee, Sh. Subhash Palekar has been proving as a viable option to address current farmers’ distress, other soil and water issues to sustain farmer’s incomes. Natural farming has inbuilt mechanisms to regenerate soil, reduce water usage, use of local available natural resources and enhance crop diversity to maintain crop quality. Dr YS Parmar University of Horticulture and Forestry, Nauni, Solan, HP, India initiated natural farming based planned principles research to establish scientific logics. The farming claims to be environmental-friendly approach than conventional agriculture, capable of attaining sustainable development objectives by using on-farm resources and reduced tillage. Introducing natural production measures to produce chemical free soils towards implementation of more resilient cropping systems through reduction in greenhouse gas emission, restoration of soil fertility and enhanced carbon sequestration. It also holds promise in reducing serious agrarian crises raised due to increase in rural indebtedness as a result of chemical-based farming. This article proposes an ecosystem-mediated farming practices to maintain a balance between social, environmental and economic pillars of sustainability.

Antimicrobial resistance (AMR) is recognised as a major healthcare problem rapidly spreading across the world. The global rise of highly resistant bacteria is well documented and closely linked to the increasing mobility of people, animals, and goods. Indeed, World Health Organization published its first-ever list of antibiotic-resistant “priority pathogens” which consists of 12 bacteria that pose the greatest threat to human health. If we do not find proactive strategies to slow down the spread of drug resistance, it is estimated that by 2050, 10 million lives per year and a cumulative 100 trillion USD of economic output will be at risk due to the rise of drug-resistant infections. The major drivers of AMR are the misuse and overuse of antimicrobial agents in both humans and animals, plus environmental pollution. Moreover, over the past three decades, it has become increasingly obvious that the majority of the novel, emerging zoonotic infectious diseases originate in animals, particularly wildlife, and that human activities, such as changes to ecosystems and land use, intensification of agriculture, urbanization, and international travel and trade, are the main drivers of their emergence. In this regard, combating AMR and emerging infections requires a One Health approach because it depends on the interrelationship between animals, people, and the environment. Recently, a novel concept to tackle AMR named “One health genomics“, was proposed. Accordingly, the Global Antimicrobial Resistance and Use Surveillance System established by the World Health Organization strongly suggests using whole-genome sequencing for surveillance of AMR to add important, policy- relevant information for AMR surveillance, to better inform strategies to tackle AMR, and to enhance the overall surveillance capacity. Implementation of the “One health genomics“ could provide an in- depth insight into the AMR landscape of a region, microbial composition across different sample sources, the genetic relatedness of the circulating multidrug-resistant bacteria, and differences in resistome profiles of interconnected sectors. In conclusion, the implementation of the One Health concept, as an integrated and unifying approach against AMR is of utmost importance at both national and global levels.

Fungal infestation is considered to be one of the main causes of decay of all types of historic works of art and source of occupational hazard for conservators, restorers and other personnel responsible for cultural heritage upkeep. Nowadays, adequate control method for suppression of fungal infestation, with minimal environmental impact, is still a considerable problem which requires a solution. Use of highly efficient inorganic and organic biocides to eradicate fungal growth, in heritage repositories and outdoor environment, is increasingly discouraged due to their toxicity, non- selective mode of action, abrasiveness, low long-term effectiveness and promotion of resistance. The need for environmentally friendly alternatives to toxic biocides has directed the research in last decade towards potential application of natural products, with new focus on beneficial bacteria due to possibility of implementing biotechnological approaches to obtain sufficient amounts of bioactive compounds. Although very limited in scope, up to date literature overview has clearly pointed out bacteria of Bacillus genus as promising green alternative for sustainable and long-term cultural heritage conservation due to their non-pathogenic nature, great physiological diversity, intensive sporulation, easy cultivation and manipulation, high metabolite yield, and ability to produce multitude of bioactive secondary metabolites, including antibacterial ribosomally synthesized lanthipeptides and antifungal non- ribosomally synthesized peptides and lipopeptides. This makes production of Bacillus-biocompounds in industrial bioreactors ideal solution for large scale in situ application on infested works of art. Conducted research has demonstrated great potential of these biocompounds to suppress growth of biodeteriogenic fungi, isolated from multitude of works of art, in in vitro experiments, with activity sometimes being comparable or even higher than commercial biocides.Several studies performed on laboratory models in simulated conditions have confirmed antifungal activity as well as absence of any form of negative impact applied bacteria-based bioformulation might have on a structural or aesthetic integrity of treated models. Additional studies are, however, necessary to determine the most adequate method of application, evaluate the long-term effectiveness, potential recolonization, and determine the costs of its production and application, before usage of bacterial biocompounds becomes groundbreaking new norm in conservation practice that replaces commercial biocides.