Bioinformatic discovery of novel collagen-binding aggregation factors in lactic acid bacteria

Lactic acid bacteria (LABs) are a unique group of microorganisms found in the diet of nearly all humans and form an integral part of the healthy human microbiome. Some LAB strains exhibit a remarkable ability for autoaggregation, mediated by Snowflake Forming Collagen Binding Aggregation Factors (SFCBAFs)—a fascinating group of proteins described so far only in LABs. To date, only five SFCBAFs have been characterized in detail: AggL from Lactococcus lactis, AggE from Enterococcus faecium, AggLb from Lacticaseibacillus paracasei, AggLr from Lactococcus raffinolactis, and AggA from Tetragenococcus halophilus. In this study, we present bioinformatically predicted novel SFCBAF candidates and demonstrate their widespread distribution among LAB species. Furthermore, we provide evidence that such proteins may not be exclusive to LABs, as homologous sequences were also identified in phylogenetically distant bacteria such as Staphylococcus aureus, Oceanobacillus spp., Bacillus spp., and others, expanding our understanding of this unique protein family

Ameliorative Effect of Banana Lectin in TNBS-Induced Colitis in C57BL/6 Mice Relies on the Promotion of Antioxidative Mechanisms in the Colon

Background: The global burden of inflammatory bowel diseases (IBDs), including ulcerative colitis and Crohn’s disease, is constantly rising. As IBDs significantly reduce patients’ quality of life, prevention and efficient treatment of IBDs are of paramount importance. Although the molecular mechanisms underlying IBD pathogenesis are still not completely understood, numerous studies indicate the essential role of oxidative stress in the progression of the diseases. Objective: The aim of this study was to investigate whether prophylactic administration of recombinant banana lectin (rBanLec) could positively affect antioxidative mechanisms in the colon and thus prevent or alleviate the severity of experimental colitis induced in C57BL/6 mice. Methods: The prophylactic potential of rBanLec, a mannose-binding lectin with immunomodulatory properties, was investigated in a model of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in C57BL/6 mice. Mice received rBanLec at various doses (0.1, 1 and 10 μg/mL) before the induction of colitis. The severity of the disease was assessed by weight loss and reduction in colon length, and correlated with histopathological findings, cytokine milieu, and oxidative stress markers in the colon. Results: The obtained results revealed that pretreatment with a low dose of rBanLec (0.1 μg/mL) significantly reduced the severity of TNBS-induced colitis, as indicated by reduced weight loss, less severe histopathological damage, and a favorable anti-inflammatory cytokine milieu (increased IL-10 and TGFβ). In addition, rBanLec pretreatment improved the activity of antioxidant enzymes (SOD, CAT, and GST) and reduced markers of oxidative stress such as nitric oxide levels at the peak of the disease. In contrast, higher doses of rBanLec exacerbated inflammatory responses. Conclusions: Our findings indicate that at low doses rBanLec can alleviate the severity of colitis by modulating oxidative stress and promoting anti-inflammatory cytokine responses, positioning rBanLec as a potential candidate for treating IBDs

Detection of peanut allergens in retail food items in Serbia (2021-2024)

Food allergies are a significant and growing public health concern globally, impacting millions of individuals and placing immense pressure on healthcare systems and personal well-being. Peanuts are one of the 14 fundamental food allergens, making them potentially harmful. As a result, the goal of this mini review is to assess the prevalence of peanuts in commercial food products in Serbia between 2021 and 2024. The current study summarizes the qualitative detection of specific DNA sequences using real-time PCR techniques on 212 commercial food goods, including frozen desserts, biscuits and related products, cream products, snacks, and sausages. Undeclared peanut allergen DNA occurred across different food categories, with positive results observed in frozen desserts (n=3), biscuits (n=4), and snacks (n=2). Peanut allergen DNA was detected in six samples with a precautionary allergen label (PAL) statement, while in the case of three samples in which peanut allergen DNA was detected, the presence of the allergen was not stated at all

Isolations and Characterization of Novel Bacterial Species from Cyprinus Carpio and Its Use in Biodegradation of Polystyrene with Emphasis on SDG 3 for Ensuring Healthy Lives and Well-Being for All Humans and Animals

Objective: This research aims to investigate the feasibility of employing beneficial bacterial species present in the gastrointestinal tract (GIT) of common carp (Cyprinus carpio) in order to biodegrade microplastics (MP), which can provide healthy lives for both humans and animals.   Theoretical framework: the main concept in this research is that the bacterial communities found in (GIT) of both people and animals can produce beneficial effects. Microorganisms that are beneficial to the host and are typically found in GIT are generally referred to as probiotics: Method: The 16S rRNA sequencing method was utilized to determine the species present in the bacterial community of Cyprinus carpio. The novel species Hafnia parallel UUNT_MP29 was utilized to conduct the biodegradation investigation. PS was used as the sole source of carbon. Discussion and the Results: During the PS batch experiment, this strain reached its highest point on the fifteenth day, displaying an incubation rate of 1.3x108 CFU/mL. Following that, the microbe entered a stationary phase, which it remained in until the experiment was finished on the 20th. According to the results, this strain appears to be a new indigenous variation capable of biodegrading PS compounds. Originality/Value: This study contributes to the existing body of knowledge by discovering bacterial species from common carp that have the potential to degrade PS. The possible application of Hafnia paralvei UUNT_MP29 in the rehabilitation of wastewater polluted by microplastics demonstrates the significance and utility of this research

Prognostic Impact of Vaccination, Comorbidity, and Inflammatory Biomarkers on Clinical Outcome in Hospitalized Patients with COVID-19

Background/Objectives: The coronavirus disease 2019 (COVID-19) has more severe symptoms and increased mortality among men than women. To address the prognostic impact of vaccination, comorbidities, and inflammatory biomarkers on classified clinical outcomes in hospitalized COVID-19 patients, we compared common and sex differences. Methods: Besides laboratory and clinical parameters at hospital admission, we performed a common and sex-based comparative analysis for the clinical outcomes, RT-qPCR analyses, and measured severe acute respiratory syndrome coronavirus (SARS-CoV-2)-specific IgM and IgG antibody levels of 702 COVID-19 patients in a single centre from June 2020 to April 2022. Results: Pro-inflammatory biomarkers (C-reactive protein (CRP), interleukin-6 (IL-6), fibrinogen, lactate dehydrogenase (LDH), D-dimer, ferritin), and liver enzymes (AST, ALT, GGT) were significantly more increased in COVID-19 male patients and generally elevated with the severity of clinical outcome, regardless of the SARS-CoV-2 variant. Cycle threshold (Ct) values of RT-qPCR testing were in negative correlation with IL-6 in COVID-19 male patients, indicating that higher viral load largely increased IL-6 levels in parallel with the severity of clinical outcome and regardless of vaccination. IgG levels were higher in early post-COVID-19 male patients. Comorbidities were more frequent in COVID-19 female patients and generally more common in the severe clinical outcomes. Vaccination was negatively correlated with the severity of clinical outcome, liver enzymes, LDH, and inflammatory parameters in hospitalized COVID-19 patients, while the risk of pneumonia was reduced. Vaccination reduced the need for corticosteroid and anti-inflammatory therapies, but increased the need for antiviral drug treatment. Conclusions: In addition to confirming inflammatory biomarkers and the importance of anti-inflammatory therapy in vaccinated patients, this study showed that vaccination reduces, but does not prevent, mortality in patients with COVID-19

Identification and Characterization of New Hafnia Strains from Common Carp (Cyprinus carpio), Potentially Possessing Probiotic Properties and Plastic Biodegradation Capabilities

Finding and characterizing new bacterial strains, including probiotic strains, is a crucial task in today’s world to expand the genetic data pool and identify new genes. In this study, we investigated the gut microbiota of one industrial species, Cyprinus carpio, and identified representatives of various microbial genera, including Citrobacter, Serratia, Bacillus, Enterococcus, and Kocuria. Notably, we discovered two strains of Hafnia with potentially probiotic properties. We conducted next-generation sequencing (NGS) of these strains, described their antibiotic resistance and antibacterial activity, and compared them with other representatives of the Hafnia genus. These strains, characterized by rapid growth, the presence of the ClpB heat shock protein gene, and genes associated with microplastic degradation, provide a promising basis for further research, including studies on their potential application in plastic biodegradation

Vaginal Bacterial Microbiome Analysis Using 16S rRNA Nanopore Sequencing Confirms Inverse Association Between Gardnerella and Lactobacillus Genera

Objective: The aim of this analysis was to explore vaginal bacterial microbiome using nanopore sequencing for the identification of species, dominance, and possible associations. Methods: Vaginal swabs were collected within one menstrual cycle, PCR was performed with modified primers 27F and 1429R. Nanopore sequencing of the 16S rRNA gene library was performed using the SQK-RBK114.96 kit (Oxford Nanopore) and a FLO-MIN114 flow cell on the MinKNOW platform (software version 24.02.8) with default parameters. Subsequent analysis was carried out using EPI2ME (version 5.2.2) with the wfmetagenomics workflow. Results: In this study, 135 vaginal microbiome samples were analyzed, collected in different menstrual phases from 35 women. The samples were divided into two groups based on the relative abundance of Gardnerella: in 46 samples it exceeded 20%. Among the samples with high Gardnerella levels, only 14 cases (30.4%) showed a Lactobacillus abundance greater than 20%, compared to 65 cases (73.0%) in the low Gardnerella group. The odds ratio calculation revealed that the likelihood of detecting Lactobacillus >20% was more than six times lower in the high Gardnerella group (odds ratio = 0.16; χ² = 28.89; p < 0.0001). Conclusions: Here we note a statistically significant inverse association between Gardnerella and Lactobacillus genera, which likely reflects competition for the same ecological niche. 16S rRNA sequencing is very useful for the quick analysis of the major species up to the genus level. Using this methodology the status of vaginal microbiota, physiological vs. pathological, can quickly be assessed without microorganism cultivation

Discrete changes in peripheral blood lymphocyte population abundance and increased TGF-β1 levels in obese women after supplementation with a novel probiotic formulation

Objective: To evaluate the combination of 7×10^10 CFU Lactiplantibacillus plantarum 299v (DSM9843), 5×10^9 CFU Saccharomyces cerevisiae var. boulardii, and 40 mg octacosanol in a randomized, double-blind, placebo-controlled trial, in particular the influence on selected immunological parameters, obese premenopausal women received one capsule daily for three months. Methods: Cell phenotyping was performed by flow cytometry, serum TGF-β1 levels were measured using a commercial reagent, while antibody levels specific for the consumed microorganisms were measured using an in-house ELISA. Measurements were performed at two time points, at t=0 and t=1. Results: In the experimental group, a significant reduction in the proportion of CD3+CD8+ cells and CD4+CD45RO+ lymphocytes was observed after 90 days (p=0.0430 and p= 0.0429, respectively). Contrary, no significant changes in the abundance of CD3+CD4+, CD3+CD56+ or CD3-CD56+ limphocytes were detected between the groups or over time. No significant difference was found in antibody levels specific to the consumed microorganisms. Elevated total TGF-β1 serum levels were detected in the intervention group at t=1 (p=0.0045), and at this time point the difference between the groups was also significant (p=0.0488). No difference in specific antibody levels was measured. Conclusions: Here we report that supplementation with a novel formulation reduced CD3+CD8+ cytotoxic cells and CD4+CD45RO+ memory T-helper cell counts, associated with low grade inflammation in the peripheral blood. We also detected an increased total serum TGF-β1, an important regulator of lean-fat body mass. The increase in TGF-β1 levels has been previously associated with probiotic consumption which should be examined with further studies

What story do antibodies tell: Population immunity of Belgrade residents as a guide for future steps

U okviru projekta „Procena imuniteta stanovnistva na vakcinama preventabilne i druge infekcije u populaciji stanovnika grada Beograda”, sprovedena je seroepidemiološka studija preseka s ciljem procene seroprevalence IgG antitela na morbili virus, mumps virus i rubela virus. Analizirani su uzorci seruma 2533 dobrovoljaca, raspoređenih u devet starosnih kategorija (1–70+ godina), primenom standardizovanih ELISA metoda. Rezultati su pokazali varijabilne nivoe seropozitivnosti u zavisnosti od vakcinalnog statusa i starosne kohorte. Ukupna seroprevalenca morbila iznosila je 74,7%, s visokim nivoima u dece (90,7%) i starijih osoba (98,4%), dok je najniža detektovana bila u grupi od 30–49 godina. Procenat seropozitivnosti na rubelu bio je 94,8%, sa ujednačenom raspodelom među svim starosnim grupama. Seroprevalenca mumpsa iznosila je 85,1%, s najnižim vrednostima kod dece uzrasta 1–5 godina (76,1%) i najvišim kod starijih ispitanika (92,6%). Rezultati ukazuju na solidan kolektivni imunitet populacije Beograda, ali i na potrebu za revakcinacijom ili dodatnim epidemiološkim merama medju pojedinim grupama, posebno u kontekstu malih boginja kod odraslih srednje dobi i zaušaka kod dece. Neophodna je kontinuirana procena efikasnosti vakcinacije i sprovođenje ciljanih imunizacionih strategija kako bi se očuvao kolektivni imunitet i smanjio rizik od izbijanja epidemija. Zaključci ove studije mogu doprineti optimizaciji nacionalnih programa imunizacije i definisanju adekvatnih javnozdravstvenih preporuka.As part of the project “Assessment of Population Immunity to Vaccine-Preventable and Other Infections in the Population of Belgrade”, a cross-sectional seroepidemiological study was conducted to evaluate the seroprevalence of IgG antibodies against measles virus, mumps virus, and rubella virus (MMR). Serum samples from 2533 voluntary participants, distributed across nine age categories (1– 70+ years), were analyzed using standardized ELISA methods. The results revealed variable levels of seropositivity depending on vaccination status and age cohort. The overall seroprevalence of measles was 74.7%, with high levels detected in children (90.7%) and older adults (98.4%), while the lowest seropositivity was observed in the 30–49 age group. The seropositivity rate for rubella was 94.8%, with a uniform distribution across all age groups. The seroprevalence of mumps was 85.1%, with the lowest values recorded in children aged 1–5 years (76.1%) and the highest in older adults (92.6%). The results indicate a solid level of herd immunity in the population of Belgrade but highlight the need for revaccination or additional epidemiological measures among certain groups, particularly regarding measles immunity in middle-aged adults and mumps immunity in children. Continuous assessment of vaccination efficacy and the implementation of targeted immunization strategies are necessary to maintain herd immunity and reduce the risk of epidemic outbreaks. The conclusions of this study may contribute to the optimization of national immunization programs and the development of appropriate public health recommendations

Shielding the gut: Can Lactobacillus rhamnosus LB-64 mitigate doxorubicin-induced enterotoxicity in a rat model?

Objective: Doxorubicin is an effective cytostatic agent, but its use is limited due to adverse effects that primarily affect the heart, kidneys, liver, and intestines. Lactobacillus rhamnosus LB-64 exhibits antagonism against pathogenic bacteria in the digestive system and is widely used as a commercial probiotic (Liobif®, Torlak). This study aims to evaluate its potential prophylactic effects on doxorubicin-induced enteropathy and malabsorption in a rat model, where these conditions are among primary causes of mortality in the acute post-treatment phase. Methods: Adult male Wistar rats (n = 36) were randomised into six groups: LIO-DOX35 (n = 6) and LIO-DOX70 (n = 6) received Liobif® (1 mL PO) for five days prior doxorubicin (5mg/0,5 mL/ kg IV), followed by continued administration until day 35 and 70 respectively. DOX35 (n = 7) and DOX70 (n = 7) received doxorubicin without probiotic supplementation. Controls received either saline (CONT; n= 5; 0,5 mL/kg, IV) or Liobif® alone (LIO-CONT; n = 7; 1mL PO). Body weight was measured weekly. Overall health was monitored daily. Results: Weight gain and healthy appearance were observed in LACT-DOX35, LACT-DOX70, CONT, and LACT-CONT groups with or without mild diarrhoea. In contrast, DOX35 and DOX70 groups exhibited weight loss and deteriorating health with profuse diarrhoea. Pathohistological analysis displayed mild bowel tissue disruption in probiotic-treated groups, whereas DOX-only groups had intestinal atrophy. Other organs revealed impairments typical for doxorubicin-treated individuals. Conclusions: Lactobacillus rhamnosus LB-65 potentially improves quality of life in doxorubicin-induced enteropathy