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Two decades of Lloviu virus in Europe: knowns and unknowns about the European filovirus
No full textIntroduction: The Lloviu cuevavirus (LLOV) was the first filovirus identified in Europe and at the temperate climate region. Its first discovery was associated with a massive die-off event of Schreibers's bats in Spain. Since its discovery this virus became the second filovirus ever isolated directly from bats. Initial in vitro experiments have suggested that this virus could be a potentially zoonotic filovirus and further experiments suggested an apathogenic nature in humans. During the last decades, several more die-off events in bats were associated with the virus, which therefore might be one of the few viruses pathogenic to bats under certain circumstances.
Methods & Materials: We initiated a complex monitoring program for Lloviu virus in 2013. During the last decade we collected and screened more than 2000 blood samples with PCR and serology from eight countries. In addition to blood samples, we screened urine, faeces, swab and ectoparasites from these animals. Using a mobile laboratory approach, we collected fresh blood samples from infected animals on-site to facilitate in vitro isolation experiments. We used the SuBK12-08 cell line for the establishment of novel isolates. In parallel we analysed the ectoparasite samples with sequencing and histology to get a better understanding about their potential vector role.
Results: We detected the viral RNA in all sample types, except oral swabs. We generated more than 20 novel viral genomic sequences, covering the whole coding region of the virus. We established four in vitro isolates. Using the sequence data, we generated throughout the years we performed molecular modelling to understand the temporal variation regarding its receptor affinity to bats and found no evidence of adaptation to better receptor binding. Additionally, we were able to calculate the molecular evolution of the virus, describe its mutational landscape and phylogenetic tree.
Discussion: Over a decade of research activities, we now possess data that allow us to evaluate the hypothesis concerning the emerging versus enzootic nature of this virus in Europe. Based on the surveillance and sequence data we believe that this virus is endemic in the whole range of this bat species. We established a hypothesis about the potential connection of die-off events, Lloviu virus infection and hibernation of these animals.
Conclusion: Although we generated significant amount of data regarding the ecology and zoonotic nature of this virus, several key questions remained to be solved. These include the investigation of potential spillover hosts, other potential bat host species and the role of ectoparasites in the natural transmission cycle. In conclusion, our efforts may lead to a deeper understanding of temperate climate filoviruses and, as a model, provide insights into the ecology and spillover mechanisms of highly pathogenic African filoviruses.Lee SS, editor. Abstracts: the International Congress on Infectious Diseases: ICID CT 2024; 2024 Dec 3-6; Cape Town, South Africa. Elsevier: 2025. p. 107752. (International Journal of Infectious Diseases; Vol. 152; Suppl.)
Comparative analysis of antioxidant enzyme activities in Lithoglyphus naticoides and Theodoxus fluviatilis: insights into redox status and environmental adaptation
No full textThe study investigates the influence of environmental conditions on redox-regulating enzymes in two freshwater snail species, Lithoglyphus naticoides and Theodoxus fluviatilis, in the Serbian section of the Danube. These species have high population densities and are characterized by their easy accessibility for sampling making them suitable candidates for biomonitoring efforts. Given the increasing concern about pollution of freshwater ecosystems, this study addresses how different concentrations of trace metals (As, Cd, Cu, Hg, Ni, Pb, Zn) at two sites — Stari Slankamen and Novi Sad — affect the antioxidant enzyme responses of snails at different periods of the year. According to the consensus-based sediment quality guidelines, both studied localities show a significant load of trace metals, especially during the summer months. Nickel concentrations are over 36 mg/kg and Zn concentrations are over 200 mg/kg. Furthermore, the NS locality has an additional contamination with Cd of over 3 mg/kg. The study measures key antioxidant enzymes—superoxide dismutase, catalase, glutathione peroxidase, glutathione
reductase, and glutathione S-transferase (GST)—as biomarkers of metal contamination. The results show
that enzyme activities are generally higher in spring than in summer, underlining the seasonal influences
on physiological responses. Preliminary results indicate that T. fluviatilis is more sensitive to environmental changes than L. naticoides, highlighting the need for species-specific considerations in biomonitoring.
Long-term studies are recommended to further investigate the chronic effects of pollution on these species
and to improve the understanding of the relationship between environmental conditions and physiological
responses in freshwater ecosystems
Changes in Soil Properties Under the Influence of Microplastics in Plastic and Open Field Production in Three Serbian Valleys
No full textCrop production in plastic greenhouses is one of the major sources of plastic pollution worldwide. The main hypothesis of this study is that the regular use of mulch film in greenhouses leads to the cumulative accumulation of microplastic particles (MPs) in the soil, which ultimately leads to changes in the soil properties. Therefore, the main objective of this study was to identify the possible changes in the physical, chemical, and biological properties of soil in greenhouses in three regions of Serbia. The following chemical parameters were determined: electrical conductivity, soil acidity, cation exchange capacity (CEC), total carbon (TC) and nitrogen (TN) content, plant-available phosphorus and potassium content, and trace element content. The following physical parameters were determined: particle size distribution, volumetric mass, specific mass, and porosity; the biological parameters that were determined were microbial respiration and labile carbon. The obtained data were processed using network analysis (NA) to identify the complex relationships between MP content and soil parameters. The NA results support the main findings that the presence of microplastics leads to the destruction of soil structure, which reduces bulk density and increases soil porosity. A strong positive correlation of MPs with soil particles < 0.02 mm and a negative correlation with CEC were found. In the Danube Valley, soil respiration was 78% higher in the open ground compared to a plastic greenhouse. The results contribute to a better understanding of the influence of MPs on soil properties and its ecological functions
Biodegradable Electrospun PLGA Nanofibers-Encapsulated Trichinella Spiralis Antigens Protect from Relapsing Experimental Autoimmune Encephalomyelitis and Related Gut Microbiota Dysbiosis
No full textPurpose: Trichinella spiralis has evolved complex immunomodulatory mechanisms mediated by excretory-secretory products (ESL1)
that enable its survival in the host. Consequently, ESL1 antigens display excellent potential for treating autoimmune diseases such as
multiple sclerosis (MS). However, whether timely controlled delivery of ESL1 antigens in vivo, as in natural infections, could enhance
its therapeutic potential for MS is still unknown.
Methods: To test this, we encapsulated ESL1 antigens into biodegradable poly (lactide-co-glycolic) acid (PLGA) nanofibers by
emulsion electrospinning as a delivery system and assessed their release dynamics in vitro, and in an animal MS model, experimental
autoimmune encephalomyelitis (EAE), induced 7 days after PLGA/ESL1 subcutaneous implantation. PLGA/ESL1 effects on EAE
symptoms were monitored along with multiple immune cell subsets in target organs at the peak and recovery of EAE. Gut barrier
function and microbiota composition were analyzed using qPCR, 16S rRNA sequencing, and metabolomic analyses.
Results: ESL1 antigens, released from PLGA and drained via myeloid antigen-presenting cells through lymph nodes, protected the animals
from developing EAE symptoms. These effects correlated with reduced activation of myeloid cells, increased IL-10 expression, and reduced
accumulation of proinflammatory natural killer (NK) cells, T helper (Th)1 and Th17 cells in the spleen and central nervous system (CNS).
Additionally, CD4+CD25hiFoxP3+ regulatory T cells and IL-10-producing B cells were expanded in PLGA/ESL1-treated animals,
compared to control animals. The migration of ESL1 to the guts correlated with locally reduced inflammation and gut barrier damage.
Additionally, PLGA/ESL1-treated animals displayed an unaltered microbiota characterized only by a more pronounced protective
mevalonate pathway and expanded short-chain fatty acid-producing bacteria, which are known to suppress inflammation.
Conclusion: The delivery of T. spiralis ESL1 antigens via biodegradable electrospun PLGA nanofiber implants efficiently protected
the animals from developing EAE by inducing a beneficial immune response in the spleen, gut, and CNS. This platform provides
excellent grounds for further development of novel MS therapies
Trichinella spiralis extracellular vesicles induce anti-inflammatory and regulatory immune responses in vitro
No full textThe helminth Trichinella spiralis, through its excretory-secretory (ES L1) products, induces immune regulatory mechanisms that modulate the host’s immune response not only to itself, but also to bystander
antigens, foreign or self in origin, which can result in the alleviation of inflammatory diseases. Under
the influence of ES L1, dendritic cells (DCs) acquire a tolerogenic phenotype and the capacity to induce
Th2 and regulatory responses. Since ES L1 products represent a complex mixture of proteins and extracellular vesicles (TsEVs) the aim of this study was to investigate the impact of TsEVs, isolated from ES L1
products, on phenotypic and functional characteristics of DCs and to elucidate whether TsEVs could
reproduce the immunomodulatory effects of the complete ES L1 product. Monocyte-derived DCs treated
with TsEVs acquired semi-matured phenotypes, characterized by low expression of human leukocyte
antigen – DR isotype (HLA-DR), cluster of differentiation (CD) 86 (CD86), and CD40, moderate expression
of CD83 and C–C chemokine receptor type 7 (CCR7), and increased expression of tolerogenic markers
indoleamine 2,3-dioxygenase 1 (IDO-1) and immunoglobulin-like transcript 3 (ILT3), together with the
unchanged production of IL-12 and IL-23, and elevated production of IL-10 and transforming growth factor (TGF)-b, compared with controls. Gene expression analysis of TsEV-treated DCs revealed elevated
levels of mTOR, Ahr, NF-jB2, RelB, SOCS1 and SOCS3, which participate in signaling pathways involved
in DC maturation and the subsequent regulation of release of both anti-inflammatory and proinflammatory cytokines. TsEVs promoted the capacity of DCs to drive polarization of Th2 and antiinflammatory responses, and impaired their capacity to induce Th1/Th17 polarization. Moreover, TsEVtreated DCs possessed a high capacity to induce conventional FoxP3 + regulatory T cells, as well as unconventional T regulatory (Tr1) cells. Tolerogenic properties of TsEV-treated DCs were retained even after
challenge with a pro-inflammatory stimulus. These findings highlight the potential of TsEVs to induce
immune tolerance, suggesting their potential use as therapeutics for the treatment of inflammatory
disorders
Biogeochemical survey of the Allchar (North Macedonia) arsenic-thallium ore body: a focus on hyperaccumulator plants
No full textBackground and Aims The Allchar site in North Macedonia has a unique geology exceptionally enriched in arsenic and thallium, making the soils extremely toxic. Surprisingly, the soils rich in these elements hosts a diverse flora, with unknown accumulation potential for most species. The aim of this study was to determine the elemental composition of plant species growing naturally in the Allchar area and to assess their relationship to the soil.
Methods Samples of the plants and associated rhizosphere soils were collected in the field and analysed for total and DTPA-extractable metal and metalloid concentrations.
Results High foliar concentrations of thallium (Tl) were found in some plant species, being the most extreme in Silene latifolia, at 79,200 µg g-1, whilst arsenic concentrations are generally low. Thallium hyperaccumulation (> 100 µg g-1) was detected in the families Violaceae, Lamiaceae and Caryophyllaceae. Particularly high foliar concentrations were found in Viola arsenica and V. tricolor subsp. macedonica, reaching 31,600 and 11,700 μg g-1, respectively. The elemental concentrations in soil and plant samples reflect the mineralogy of the three different areas investigated at Allchar, with the highest mean values for thallium and arsenic at Crven Dol, and 249 and 3970 μg g-1, respectively, in plants analysed.
Conclusion The present study led to the discovery of new thallium hyperaccumulating plant species, such as Clinopodium alpinum, Anthyllis vulneraria and Linum hirsutum, whereas the thallium concentrations found in Silene latifolia are the highest known in nature
Temperature related changes in wing size and shape in two sibling species, Drosophila melanogaster and D. simulans (Diptera: Drosophilidae)
No full textMorphological and life-history traits represent characteristics influenced by
various environmental factors that can contribute to adaptive variability in many organisms.
Temperature is an important environmental factor that drives phenotypic change in different traits in
Drosophila (developmental time, fertility, viability, body size, pigmentation, etc.). Therefore, the aim of
this study was to investigate the effect of temperature on wing size and shape in two sibling species,
Drosophila melanogaster and D. simulans.2nd International Electronic Conference on Entomology (IECE 2025); Online; 2025 May 19-21. Basel: MDPI; 2025. p. 23
Sexual dimorphism in size and shape of morphological traits in Drosophila simulans (Diptera: Drosophilidae): geometric morphometric approach
No full textSexual dimorphism (SD) in different morphological traits (both non-sexual and secondary sexual
traits), colour pattern, ornamentation, and behavioral repertoire has been analyzed in different animal
species. It can be defined more precisely when its two different components, sexual size dimorphism
(SSD) and sexual shape dimorphism (SShD), are analyzed separately. In spite of the fact that SSD
and SShD are equally relevant components of sexual dimorphism, SShD has not been the subject of
a large number of zoological studies. Both SSD and SShD in some morphological traits have been
poorly analyzed in Drosophila simulans. Therefore, the aim of the present study was to investigate
the presence of SSD and SShD in three morphological structures (the head, the wing, and the leg) in
D. simulans. To analyze SSD and SShD in the abovementioned morphological traits, the geometric
morphometric approach was applied.2nd International Electronic Conference on Entomology (IECE 2025); Online; 2025 May 19-21. Basel: MDPI; 2025. p. 22
Preparation and analysis of phytopreparations from the walnut septum biomass
No full textRecently, foreign researchers have been actively working on studying Juglans regia L. septum, which indicates their rich phytochemical composition and a wide range of biological properties. It has been shown that extracts from walnut septum have high antioxidant, antitumor, and antiflogistic activity, as well as antimutagenic, regenerating, and bactericidal potential. The work aimed to obtain a natural complex of phenolic compounds from the biomass of walnut septum of selected varieties to study the phytochemical composition of the obtained phytopreparations and their antioxidant properties. To extract phenolic compounds, samples of the biomass of walnut septum were mixed with an extraction mixture in a ratio of 1:30 by weight and subjected to ultrasonic treatment at 100 W with a frequency of 20–25 kHz for 10 minutes. The resulting aqueous ethanol extracts were condensed under vacuum, and their physicochemical characteristics and antioxidant properties were determined. The identification and analysis of phenolic compounds in the studied phytopreparations was done using the HPLC-MS/MS method. Seventeen phenolic compounds were found in the composition of the obtained phytoextracts, most of which belong to flavonoids. It was found that the total content of identified flavonoids and their glycosides in aqueous ethanol extracts obtained from the biomass of Juglans regia L. septum was 13.54 mg/g in a thick extract of phenolic compounds – 119.75 mg/g. The total content of the identified phenolic acids in aqueous-ethanol extracts from the biomass of the septum was 4.22 mg/g, in the thick extract – 69.43 mg/g. The antioxidant activity of the studied phytopreparations amounted to 94% inhibition of the DPPH radical. The results of the analysis and identification of the phytochemical composition indicate that the obtained phytopreparations from the biomass of walnut septum are a valuable source of phytomicronutrients as part of functional food ingredients and can also be considered pharmaceutical substances for therapeutic use
Chemical Profiling and In Vitro Evaluation of Bioactive Properties of Evernia prunastri Extract: Implications for Therapeutic Applications
No full textEvernia prunastri (L.) Ach. (Parmeliaceae), an edible lichen commonly known as oakmoss, was traditionally used by Egyptians to make bread. In this study, the ethyl-acetate (EtOAc) extract of E. prunastri was investigated for its potential therapeutic applications in diabetes mellitus, Alzheimer’s and Parkinson’s diseases, oxidative stress, and bacterial infections. The extract exhibited significant in vitro enzyme inhibition activities, including anti-amylase and anti-glucosidase activities linked to diabetes and anti-cholinesterase and anti-tyrosinase activities associated with Alzheimer’s and Parkinson’s diseases. The antioxidant activity was evaluated through multiple assays, including free radical scavenging (DPPH and ABTS), reducing power (CUPRAC and FRAP), metal chelation, and phosphomolybdenum methods, demonstrating strong oxidative stress relief potential. The antibacterial properties were also confirmed through antibacterial testing, showing efficacy against a range of bacterial strains. Total phenolic and flavonoid contents were quantified, while the chemical profile of the EtOAc extract was determined by LC-HRMS/MS analysis. The chemical composition was predominantly characterized by depsides (evernic acid and atranorin), phenolic acids (orsellinic acid), and dibenzofurans, revealing a diverse array of bioactive secondary metabolites. The extract demonstrated a broad spectrum of biological activities, including enzyme inhibition, antioxidant effects, and antibacterial properties. This study highlights the potential of E. prunastri as a functional food, providing a rich source of bioactive compounds with numerous health-promoting effects, and it suggests its relevance in therapeutic applications for chronic diseases such as diabetes, Alzheimer’s, and bacterial infections