Beer is a widely consumed carbonated beverage made from natural ingredients, including malted cereal, hops, yeast and water. It is rich in nutrients and contains carbohydrates, minerals, vitamins, amino acids and polyphenols. The brewing process, however, generates a significant amount of solid waste, including hot trub, a slurry of entrained wort, hop particles and mainly unstable high-molecular-weight colloidal proteins that coagulate during the boiling of the wort. Given the environmental impact of agro-industrial waste, finding sustainable methods to reuse this waste by transforming it into bio-products is crucial.

The aim of this study was to assess the potential of converting these by-products into biologically active extracts suitable for use as functional ingredients in cosmetic and pharmacological formulations. The extracts consisted of one alcoholic and one hydroalcoholic extract prepared by Soxhlet extraction and one hydroalcoholic extract prepared by maceration. The extracts' antibacterial activity against the Gram-positive bacteria S. aureus, S. aureus (MRSA), S. epidermidis, B. cereus, E. faecalis, S. mitis, S. pyogenes, and S. mutans, as well as against the Gram-negative bacteria E. coli and P. aeruginosa and against the yeast C. albicans, was determined by the agar diffusion method and the minimum inhibitory concentration (MIC) by the microdilution method. The minimum inhibitory concentration and inhibition zones values obtained showed significant inhibitory effects against the Gram-positive bacteria tested, with the Soxhlet extracts presenting the best inhibitory results for both hydroalcoholic and alcoholic extracts. In both methods, the extracts showed the best antibacterial activity against the microorganisms S. epidermidis and B. cereus, with an MIC of 0.63 and 0.31 mg/mL, respectively. The tested extracts seem to be promising, low-cost antibacterial agents that can be incorporated in cosmetic and pharmacological formulations. Additional studies will be conducted to assess their antioxidant activity and safety by using in vitro and in vivo models and investigating the chemical composition of the extracts.

Despite tremendous progress in the prevention and treatment of fungal infections, invasive fungi like Candida species remain the major cause of morbidity and mortality worldwide [1]. In fact, the Global Action Fund for Fungal Infections estimates that each year, almost 300 million individuals contract a fungal infection, and over 1.5 million of them died as a result [1,2]. Candida can enter the bloodstream and spread to internal organs, but it can also commonly cause surface infections, like those in the skin or mucous membranes, which can be readily and effectively treated [3]. This fungal infection has been observed in high-risk patients, such as those receiving allogeneic stem-cell transplants and those receiving high-dose chemotherapy for acute leukemia [4]. These patients are more vulnerable to infections since their immune systems are weakened during the transplant process. Systemic fungal infections are still difficult to diagnose. Therefore, developing early diagnosis methods that are more precise, sensitive, and effective is required. A quick, easy, and accurate method for identifying fungal infections in patients following hematopoietic stem cell transplantation (HSCT) was devised in this study. To overcome this problem, an electrochemical nanogenosensor for the detection of Candida albicans was developed. This nanogenosensor was assembled in an innovative low-cost electrochemical paper-based analytical device (ePAD). The electrochemical signal's sensitivity was increased through the application of a sandwich hybridization procedure. Preliminary findings indicate that these nanogenosensors can be used to identify Candida spp. in a synthetic fungus sample. Despite these findings, work is being performed to enhance the nanogenosensor for the quantification of Candida albicans, and this procedure will be verified through more study. The application in a hospital context will be covered in future studies in terms of sensitivity, accuracy, response time, issues, and potential.

Introduction: Understanding COVID-19 pathogenesis and immune responses relies on robust in vitro systems that utilize purified SARS-CoV-2 virions. This study focused on developing a rapid, benchtop purification method for SARS-CoV-2, designed to eliminate non-viral substances and cytopathic agents, enabling immunological experiments.

Methods: The SARS-CoV-2 delta variant was propagated in Vero E6 cells within a BSL-4 laboratory. Complete virus inactivation was achieved using UV irradiation and was confirmed via standard virological assays. The purification procedure involved filtration through a 0.22 μm membrane, followed by anion exchange chromatography, with desalting performed as an optional step. PBMCs were isolated from healthy donor blood using SepMate™ tubes with slight modifications to the manufacturer’s protocol. These PBMCs were incubated with either purified or unpurified virus, and cells and immunological interactions were analyzed through flow cytometry.

Results: The purification method successfully removed 99% of non-viral proteins, leaving primarily viral proteins. RT-PCR and rapid antigen tests demonstrated an approximately 70% recovery of viral RNA. Flow cytometry confirmed that the unpurified virus caused significant, almost total, CD14+ monocyte destruction. In contrast, monocytes incubated with purified virus maintained viability, indicating reduced cytotoxic effects.

Discussion: This streamlined purification process provides a reliable, rapid tool for preparing SARS-CoV-2 for immunological studies. While anion exchange chromatography was highly effective, observed discrepancies between PCR and rapid antigen test results after desalting merit further exploration. Future studies with active viruses will help determine the infectivity of purified preparations.

Acknowledgment: This research was conducted in collaboration with the National Laboratory of Virology and the Flow Cytometry Core Facility of the University of Pécs. The project was supported by grants 2020-2.1.1-ED-2020-00100 and RRF-2.3.1-21-2022-00010 from the National Research, Development, and Innovation Office of Hungary.

The widespread use of polyethylene (PE) in industries such as agriculture and construction poses environmental challenges due to its recalcitrance in natural ecosystems. This study focuses on the microbial and physicochemical interactions of soils from Colombia, in the degradation of buried polyethylene pipes, combining molecular, microbiological, and physicochemical approaches to understand soil–microbe–plastic dynamics. Soil samples were collected from sites with buried polyethylene pipes exhibiting degradation signs. Microbial communities were characterized using 16S rRNA and ITS sequencing, with Nanopore technology, to identify bacterial and fungal taxa. Additionally, microbiological analysis was performed by taking swabs from the surface of degraded pipes to directly observe microbial colonization. Fungal isolates were cultured on Potato Dextrose Agar (PDA) to correlate traditional isolation techniques with molecular findings. Soil physicochemical properties, including pH, moisture, redox potential, particle size distribution, and others, were analyzed. Polyethylene degradation was assessed through surface analysis by scanning electron microscopy (SEM). Bacterial phylum Proteobacteria and genera such as Pseudomonas, along with fungal genera such as Aspergillus and Fusarium, were identified as key players in soils with higher degradation rates. Microbiological analysis of pipe surfaces confirmed microbial colonization, consistent with soil sequencing data. Fungal isolates grown on PDA matched molecular findings, validating the integration of traditional and molecular approaches. SEM imaging revealed significant surface erosion and microbial colonization on the polyethylene pipes. Physicochemical analyses showed that soils with neutral pH and higher moisure content supported more active and diverse microbial communities. This study demonstrates the critical role of microbial activity and soil physicochemical properties in polyethylene degradation. The integration of microbiological, molecular, and physicochemical methods offers a comprehensive framework to advance sustainable strategies for managing plastic pollution in soil environments.

In the food industry, synthetic preservatives are commonly used to inhibit microbial spoilage, prevent chemical and nutritional changes, and extend shelf life. However, growing consumer demand for healthier, minimally processed foods without synthetic additives has lead toto a search for natural preservation alternatives, such as antimicrobial peptides from yeasts. Pathogenic microorganisms such as Escherichia coli, Listeria monocytogenes, and Salmonella spp. represent major contamination risks to food products. To address these risks, various bioactive compounds with antimicrobial properties, including antimicrobial peptides, bacteriocins, and mycocins, have been identified as potential natural preservatives. This study aimed to assess the antimicrobial efficacy of peptide fractions from Saccharomyces cerevisiae, Wickerhamomyces anomalus, and Tetrapisispora phaffii against E. coli (ATCC 25922), L. monocytogenes (ISA 4008), and Salmonella spp. (ISA 4008). MIC tests revealed varying levels of activity, with peptides from S. cerevisiae demonstrating the strongest effect, achieving an MIC of 250 µg/mL against all three pathogens. Peptides from W. anomalus showed MICs of 2300 µg/mL for both E. coli and L. monocytogenes, and 1150 µg/mL against Salmonella spp. In contrast, peptides from T. phaffii required 1250 µg/mL to inhibit E. coli and Salmonella spp., and 2500 µg/mL to inhibit L. monocytogenes, indicating differing levels of antimicrobial efficacy. Additionally, a challenge test was conducted to determine the shelf life of the juice. The objective was to assess the antimicrobial efficacy of yeast-derived peptides for preserving watermelon juice at 25°C. The results showed that S. cerevisiae peptides achieved a >5-log CFU/mL reduction in E. coli within seven days, reducing the pathogen to undetectable levels and suggesting irreversible cellular damage. Peptides from W. anomalus also exhibited antimicrobial effects, though less pronounced, with a reduction of 1.1 log CFU/mL. These findings suggest that yeast-derived peptides, particularly from S. cerevisiae, could serve as effective natural biopreservatives to enhance the microbial safety of fruit juices.

Composting is an ancient and sustainable method of recycling organic waste into humus, a nutrient-rich substance that enhances soil productivity and reduces the environmental impact of waste disposal. Microorganisms, including fungi and bacteria, are essential to this process, as they drive the decomposition of organic matter.

This research aimed to identify fungal and bacterial species involved in composting and optimize their roles in decomposition. Compost samples were collected from Lusófona University's composting system after six months of processing organic waste, primarily fruit peels and vegetable scraps mixed with dry plant material. Microbiological analyses, including DNA extraction, PCR amplification, and Sanger sequencing, were used to identify the microorganisms present.

The results showed that the most abundant microorganisms were filamentous fungi, with the most predominant species being Aspergillus heyangensis, Aspergillus creber, Cladosporium asperulatum, Pestalotiopsis lespedezae, and especially Penicillium brevicompactum, which was the most prevalent. These species dominate the decomposition of the complete residue and have potential applications in industry and composting. The bacterial species identified were predominantly represented by the genera Bacillus (e.g., B. halotolerans, B. subtilis, B. amyloliquefaciens, B. mojavensis), as well as species from Sphingobacterium (S. kitahiroshimense), Serratia (S. fonticola), and Pseudomonas (P. fluorescens). The collective contribution of these microorganisms to the breakdown of organic material during composting is significant.

This study highlights the diversity and significance of fungal and bacterial communities in composting, emphasising their potential to optimize organic waste recycling processes. Understanding the microbiota involved in composting opens avenues for developing biotechnologies, such as microbial inoculants, to enhance process efficiency and sustainability. Future research should focus on optimizing environmental conditions to support beneficial microorganisms, further advancing composting practices and producing higher-quality humus.

Background: Middle ear infection (otitis media) remains the most common childhood disease diagnosed among Australian Indigenous children. Despite extensive antibiotic usage, it persists as a significant health concern, characterized by substantial morbidity and economic burden. Streptococcus pneumoniae, non-typeable Haemophilus influenzae, and Moraxella catarrhalis are the most common culprits responsible for acute otitis media. Currently, pneumococcal and influenza vaccines are utilized as a preventive measure against otitis media. However, their efficacy is limited to prevent carriage and/or illness induced by non-vaccine serotypes. Otitis media stemming from non-vaccine serotype pneumococci, non-typeable H. influenzae, and M. catarrhalis continue to pose significant healthcare challenges.

Methods and Findings: Given the expansive heterogeneity of pathogens involved in otitis media, a multicomponent vaccine is essential to confer adequate protection. This study is focused on devising a novel multicomponent vaccine targeting two major etiological agents, H. influenzae and M. catarrhalis. Our vaccine formulation features novel protein antigens derived from M. catarrhalis, non-typeable H. influenzae, and genetically modified outer-membrane vesicles (OMVs) from M. catarrhalis. We successfully expressed and purified recombinant protein antigens from NTHi and M. catarrhalis using E. coli expression system and immobilized metal affinity chromatography. We also developed knockout mutants of M. catarrhalis lacking targeted immunodominant strain-variable proteins. These mutants are currently being used to purify OMVs. Preclinical investigations will encompass a rigorous evaluation of vaccine immunogenicity and cross-reactivity against heterologous strains in mice.

Conclusion: By combining novel protein antigens with genetically modified outer-membrane vesicles, this study aims to overcome the limitations of current vaccine approaches. Forthcoming preclinical evaluations will provide critical insights into its immunogenicity and cross-reactivity, paving the way for future advancements in otitis media prevention.

Introduction: Dogs have been perceived as providing companionship to all members of the family, yet they can act as a vector for the transmission of parasitic agents to humans, particularly those who own or care for them. Traditional medicine seem to demonstrate effects in the treatment of diseases in animals, however, it is necessary to prove their effectiveness using more recent scientific evidence.

Objective: The objective is to identify medicinal plants commonly used in the treatment and prophylaxis of parasitic infections in dogs.

Methods: A literature review was conducted by gathering data from the MEDLINE database, using the following search equations: “(“Plants”[Mesh] OR (“Plants/parasitology”[Mesh]) AND (“Phytotherapy/veterinary”[Mesh])”; “Medicinal Plants Treatment for Infested Dogs”; “Parasitic Infections in Dogs Treatment with Plants”. The selection criteria established ensured that only articles published between 2000 and 2023, which referenced the utilisation of medicinal plants in the management and prevention of parasitic infections in canines, were included. Furthermore, review articles and articles that referenced pharmacological treatments or treatments for other comorbidities were excluded.

Discussion and Conclusion: A total of 26 articles were selected, resulting in the identification of 112 species from 16 countries that can be used to treat or prevent infections and parasitic infestations. The most commonly used medicinal plants were: Azadirachta indica, Juniperus communis L., Melissa officinalis, Clibadium surinamense L., Olea europaea L., Juglans regia L., Allium sativum L., Coriandrum sativum L., Artemisia cina Berg ex Poljakov, Calendula officinalis L., Mentha piperita L., Cocos nucífera L., Ricinus communis L., Andrographis paniculata (Burm.f.) Wall. ex Nees and Juglans nigra L. The species in question contain a variety of compounds that may confer medicinal properties; however, the anthelmintic and anti-parasitic activity may be attributed to the presence of phenolic compounds, which are known to posses a range of beneficial properties, including anti-parasitic and anthelmintic effects.

Leeches (Hirudinea), a diverse group of annelids, are primarily classified as either parasites or predators, depending on their feeding methods and life strategies. These groups differ in habitat and dispersal. This diversity plays a significant role in the functioning of aquatic ecosystems, especially in the context of epidemiological threats posed by the transmission of pathogenic fungi by leeches. Due to fungi's ability to adapt rapidly to new environments and given their dissemination through various vectors, including leeches, these diseases can spread quickly. This study examined the mycobiome of the parasitic leech Glossiphonia complanata, which has a wide range of hosts, making it a potentially effective vector for the spread of fungal diseases.

Leeches were collected from five water reservoirs located on the Morasko Campus of Adam Mickiewicz University. Anesthetized leeches were surface-sterilized with 75% ethanol, dissected into fragments, and placed on PDA medium. DNA was extracted from the cultivated fungal colonies, followed by PCR amplification of the ITS region sequences. The PCR products were sequenced, and additional morphological analyses of the fungi were conducted.

In addition to widely recognized environmental fungi such as Cladosporium spp., the fungal species Papiliotrema aurea was detected and identified within the bodies of the leeches. This species, previously isolated from the Chinese mitten crab (Eriocheir sinensis), exhibits a notable capacity for colonization in various organisms. Such findings imply that P. aurea not only possesses the ability to thrive in specific microenvironments but also raises concerns regarding its potential pathogenicity.

In summary, the ecological diversity of leeches and their ability to interact with other organisms, including pathogenic fungi, highlight their importance in aquatic ecosystems. Research on the mycobiome of leeches provides valuable insights into potential vectors of fungal diseases. Understanding these mechanisms can contribute to a better management of epidemiological risks and the protection of aquatic ecosystems from pathogen invasions.

Staphylococcus aureus is a major pathogen responsible for numerous human and animal diseases, including Staphylococcal scalded skin syndrome, folliculitis, and mastitis. Known for its virulence factors, it can evade host immune responses and enhance colonization. Recent research has focused on alternative in vivo models, such as the Galleria mellonella larval model, which offers a cost-effective, less harmful, and accessible approach to studying pathogens and testing therapies. This model is compatible with the 3R principle (replacement, reduction, and refinement).

In this study, the virulence of 35 S. aureus strains isolated from humans, dogs, and cats was assessed both in vivo and in vitro. The bactericidal effect of a commercial human serum reagent on these strains was evaluated, and selected isolates were tested in the G. mellonella model. Biofilm formation was also analyzed using the crystal violet assay. Most strains were resistant to the human serum reagent, with only three being susceptible. In the larval model, varying levels of virulence were observed. Biofilm formation was detected in only five out of thirty-five strains.

No significant differences in serum resistance were observed between strains from different hosts. The Gram-positive structure of S. aureus likely explains this. Strains exhibiting the MRS (Methicillin-resistant Staphylococcus) phenotype caused higher mortality in the larvae, and those capable of forming biofilms displayed both low and high virulence levels in vivo on the larval model.

This study was partially financed by an internal grant of the University of Wrocław IDUB titled "Galleria mellonella - the use of larvae in the optimization and improvement of the in vivo insect model for bacterial pathogenicity studies.", No. BPIDUB.7.2024

The tested strains were graciously provided by the Wrocław University of Environmental and Life Sciences, Department of Epizootiology and Clinic of Birds and Exotic Animals.