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Shielding the gut: Can Lactobacillus rhamnosus LB-64 mitigate doxorubicin-induced enterotoxicity in a rat model?
No full textObjective: 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
Detection of peanut allergens in retail food items in Serbia (2021-2024)
No full textFood 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
Ameliorative Effect of Banana Lectin in TNBS-Induced Colitis in C57BL/6 Mice Relies on the Promotion of Antioxidative Mechanisms in the Colon
No full textBackground: 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
A novel type of autoaggregation in lactic acid bacteria promoted by new AggS aggregation factor from Streptococcus thermophilus CC40-4S
No full textAutoaggregation in lactic acid bacteria is considered a beneficial probiotic trait and can be used in the food and medical industries to enhance the properties of utilized microorganisms. Currently, a group of aggregation-promoting factors (APFs) in lactic acid bacteria, known as Snowflake Forming Collagen Binding Aggregation Factors (SFCBAFs), is well described. These are large proteins with a molecular mass of over 170 kDa, containing collagen-binding domains and a repeat region, forming a unique autoaggregation phenotype. Here we describe a new type of autoaggregation in lactic acid bacteria found in Streptococcus thermophilus CC40-4S. The whole genome of the autoaggregation-positive strain S. thermophilus CC40-4S was sequenced, and bioinformatic analysis predicted a putative gene aggS involved in autoaggregation, located on the chromosome and flanked by insertion sequences. The aggS gene disruption by homologous recombination using the temperature-sensitive vector pSC led to the loss of the aggregation phenotype. Cloning and heterologous expression in Lactococcus lactis subsp. cremoris MG1363 confirmed the role of the AggS protein in autoaggregation, given that a strong aggregation phenotype was obtained. Like SFCBAF-type APFs, AggS is a large protein (237 kDa) with a repeat region, but it does not contain collagen-binding domains and forms an autoaggregation phenotype with small aggregates. To our knowledge, this is the first report on a gene coding for an aggregation-promoting factor in S. thermophilus
Vaginal Bacterial Microbiome Analysis Using 16S rRNA Nanopore Sequencing Confirms Inverse Association Between Gardnerella and Lactobacillus Genera
No full textObjective: 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
Cathepsin L inhibition enhances microglial CX3CR1 expression in EAE
No full textCathepsin L (CatL), a lysosomal cysteine protease, is highly expressed in antigen-presenting cells and
plays a role in the processing of autoantigens. It is widely expressed throughout the CNS and is associated with microglia-driven neuroinflammatory responses. Previous studies suggest that inhibition
of catL activity may offer therapeutic potential for the treatment of multiple sclerosis. The aim of this
study was to investigate the effects of a selective catL inhibitor [CC(=O)c1cccc(NC(=O)c2cnc3sccn3c2=O)
c1] on microglia in mice immunized for experimental autoimmune encephalomyelitis (EAE).
The catL inhibitor was administered intraperitoneally for five consecutive days from the onset of clinical signs of disease. Mononuclear cells were isolated from the spinal cord (SC) of EAE rats and analysed
by flow cytometry. Given that this was the first in vivo application of this inhibitor, hepatotoxicity and
nephrotoxicity were also investigated. The administered catL inhibitor reduced the infiltration of the
SC by CD4+ T lymphocytes. On the other hand, the proportion of CD11b+ cells among the mononuclear
cells isolated from the SC was higher in the treated EAE mice. CatL inhibitor decreased the proportion
of CD45high cells among CD11b+ cells, most likely monocyte-derived macrophages that infiltrated the
SC and increased the proportion of CD45low/int cells among CD11b+ cells, which correspond to microglia. The proportion of neuroprotective CX3CR1+ cells among microglial cells was significantly higher in treated mice than in untreated EAE mice. These findings reinforce the hypothesis that catL could
serve as a potential therapeutic target for treating multiple sclerosis
Novel autochthonous strains from Cyprinus carpio as candidates for probiotic use and microplastic-degrading properties
No full textIn the modern era, identifying and characterizing novel bacterial strains with possible probiotic potential and environmental bioremediation capabilities is an emerging focus in microbiology and biotechnology. This study analysed the cultivable gut microbiota of the freshwater fish, Cyprinus carpio, and identified six different bacterial genera, including Citrobacter, Serratia, Bacillus, Enterococcus, and Kocuria. Among these, two novel autochthonous strains—Hafnia alvei UUNT_MP41 and Hafnia paralvei UUNT_MP29—were isolated and selected for further investigation due to their promising probiotic traits and potential to degrade microplastics in aquatic ecosystems. Both strains were evaluated for antibacterial activity against pathogens and susceptibility to a broad spectrum of antibiotics. Whole-genome analysis using next-generation sequencing (NGS) revealed the presence of genes potentially associated with probiotic properties, such as ClpB, as well as genes potentially involved in the biodegradation of common microplastics, including the tesA gene, a homolog of the PpEst gene from the genome of Pseudomonas pseudoalcaligenes, and the lipR gene, a homolog of the EstC9 gene from the genome of Acidocella sp. Here, we performed a more in-depth analysis of the similarity between the genes/proteins we identified as potentially involved in plastic biodegradation and previously described ones. Notably, the identified strains' potential to degrade microplastics under conditions relevant to the human gastrointestinal system positions them as candidates for a new generation of dual-function probiotics, supporting both human health and microplastic detoxification. These findings lay the groundwork for future development of multifunctional probiotic formulations with environmental and therapeutic benefits
Mpox Cases in Serbia, 2022
No full textBackground: On 23 July 2022, the World Health Organization (WHO) declared the mpox multi-country outbreak as a Public Health Emergency of International Concern. This study aimed to identify the epidemiological and clinical characteristics of confirmed mpox cases reported in Serbia in 2022. Methods: The mpox WHO case definition was used. Incidence rates (IRs) and incidence rate ratios (IRRs) by age groups and nomenclature of territorial units for statistics level 3 (NUTS-3) with 95% confidence intervals (CIs) were calculated. Results: Between June and October 2022, 43 laboratory-confirmed cases were reported. All were unvaccinated males, with the mean age of 34 (±7.4) years. Out of the total, 72.1% cases were men who have sex with men (MSM), who reported sexual intercourse either with multiple or unknown partners (p < 0.01). Fifteen cases (34.9%) lived with HIV, mostly in the 30–39 age group (p = 0.023). People living in Belgrade City NUTS-3 were six times more likely to become infected compared to South Backa citizens (IRR: 6.03, 95% CI: 1.47–25.53). Conclusions: In Serbia, mpox mainly affected MSM aged 30–39 and living in urban areas. Health promotion and vaccine implementation should be prioritized in populations with a higher risk
Next-Generation Pertussis Vaccines
No full textPertusis je respiratorna bolest koja se može sprečiti vakcinacijom, a izaziva je Bordetella
pertussis, prisutna u ustima, nosu i grlu zaražene osobe. Kako su ljudi jedini rezervoar ove bakterije,
potpuna vakcinacija protiv pertusisa i visoka stopa imunizacije od izuzetnog su značaja, naročito
imajući u vidu da su odojčad mlađa od godinu dana najugroženija od teških oblika bolesti i smrtnog
ishoda. Vakcinacija celim ćelijama (engl. WCV – whole-cell vaccine) sadrži neaktivne bakterije, dok
acelularna vakcina (engl. ACV – acellular pertussis vaccine) sadrži proteinske komponente Bordetella
pertussis, kao što su inaktivisani pertusis toksin, filamentni hemaglutinin, pertaktin i fimbrije.
Acelularna vakcina razvijena je kao odgovor na izveštaje o neželjenim reakcijama koje su se javljale
nakon primene vakcine sa celim ćelijama u određenim zemljama. Uvođenje acelularne vakcine tokom
90-ih godina prošlog veka dovelo je do postepenog porasta broja slučajeva pertusisa. WCV vakcina
izaziva snažniji imunološki odgovor, sličniji prirodnoj infekciji, i pruža dugotrajniju zaštitu. U zemljama
koje koriste ACV preporučuje se pojačan nadzor, kao i uključivanje dodatnih doza (buster doza).
Posebna pažnja se posvećuje vakcinaciji trudnica u cilju zaštite novorođenčadi. Vakcine koje sadrže
rekombinantni, genetski detoksifikovani pertusis toksin nedavno su registrovane, a njihova prednost
je što omogućavaju manju količinu antigena po dozi za postizanje zaštite. Ipak, postoji potreba za
razvojem nove generacije pertusis vakcina koje bi zadržale prednosti vakcina sa celim ćelijama – kao
što su indukcija adekvatnijeg imunološkog odgovora i dugotrajna zaštita – o čemu će ovde biti reči.Pertussis is a vaccine-preventable, respiratory disease caused by Bordetella pertussis, present
in the mouth, nose, and throat of an infected person. As humans are the sole reservoir, complete
vaccination against pertussis and high vaccination coverage is of utmost importance, especially as
infants aged <1 year are at greatest risk of serious disease and death. Whole-cell pertussis vaccine
(WCV) contains nonviable bacteria and acellular pertussis vaccine (ACV) - contains protein
components from B. pertussis such as inactivated pertussis toxin, filamentous hemagglutinin,
pertactin, and fimbriae. The acellular vaccine was developed in response to reports of adverse
reactions upon administering the whole-cell vaccine in certain countries. The introduction of acellular
vaccine in the 90s has led to a lower level gradual increase of the number of pertussis cases. The WCV
vaccine stimulates a strong immune response more similar to infection and protection which is more
durable. For countries that have adopted ACV increased monitoring is advised as well as the inclusion
of booster doses. Special focus is given on the vaccination of pregnant women to protect the
newborns. Recombinant, genetically detoxified pertussis toxin containing vaccines have been recently
registered the benefit being that this reduces the amount of antigen needed per dose to confer
protection. Nevertheless, a need exists for the development of a new generation of pertussis vaccines,
which will have the benefits of whole cell vaccine, such as the induction of more adequate immune
response and durability of protection, which will be discussed here
What story do antibodies tell: Population immunity of Belgrade residents as a guide for future steps
No full textU 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