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The aim of this study was to evaluate the antibacterial activity, quality and stability of creams (at 1 % concentration) prepared with synthesized silver(I) complexes: [Ag(Nam)2]NO3·H2O ( AgNam), [Ag2(HGly)2]n(NO3)2n (AgGly) (Nam – nicotin-amide, Gly – glycine) and silver(I) sulfadiazine (AgSD), which is commercially available. Antibacterial activity was evaluated by agar well diffusion method and in in vivo case. The pure silver(I) complexes as well as all three tested creams loaded with AgGly, AgSD and AgNam showed antibacterial potential. Moreover, the creams loaded with AgGly and AgNam showed higher antibacterial effects against S. aureus and B. subtilis than the cream loaded with AgSD. In terms of appearance, all cream samples were opaque and odourless, and no phase separation was observed. Creams were soluble in water (o/w emulsions) and they had a pseudoplastic behaviour. The pH of the creams was in the range of 4.87–5.75. No visible changes were observed in the case of commercially used AgSD cream during one month testing period at conditions –16 ± 1 °C; 6 ± 1 °C and 56 % relative humidity; 20 ± 1 °C and 58 % relative humidity and 40 ± 1 °C and 75 % relative humidity. However, creams containing AgGly and AgNam changed their colour depending on the tested conditions.

The flow cytometry method is used in many fields, not only scientific, but also clinical. In science, flow cytometry is used in immunology, molecular biology, microbiology or plant biology. In medicine, has its use, in the diagnosis of tumours, in reproductive and prenatal diagnosis, in transplants or in haematology. In our experimental work, we used this method to analyse colostrum and milk of sows. The aim of this study was to determine the number of somatic cells, the number of leukocytes and the number of T-lymphocytes. Colostrum samples were collected within 24 hours of delivery and then milk samples were collected at weekly intervals during 3 weeks. Statistical analysis revealed significant differences in the number of leukocytes (CD45+) in colostrum and milk. The most significant difference was noted between colostrum and milk in weeks 2 and 3 of the experiment (P < 0.0001), with the highest values found in colostrum. On the other hand, we found that the levels of helper T-cells (CD4+) and cytotoxic T-cells (CD8+) gradually increased over time (P > 0.05), with the highest values observed in the third week after farrowing. We can conclude that flow cytometry can be successfully used for the examination of somatic cells in sows’ milk.

American foulbrood is amongst the most dangerous diseases of the bee-family affecting many honeybee colonies worldwide. In countries of European Union, based on veterinary legislation, the bee colonies tested positive to American foulbrood are eradicated with high economic losses. It is therefore necessary to look for effective prevention, especially by the using of natural ingredients such as probiotics. In this study, we used lactobacilli isolated from digestive tracts of adult healthy honey bees and selected based on their good probiotic properties and ability to inhibit the growth of Paenibacillus larvae. These isolates were identified as Lactobacillus brevis and Lactobacillus plantarum. Night cultures of both strains were used for the preparation of probiotic suspensions and pollen was selected as an appropriate carrier for application of probiotic lacto-bacilli to the hives. Half a litre of pollen suspension was prepared for each hive. The suspension for experimental hives contained probiotic lactobacilli in concentration of 107–108 CFU.ml−1. Bees in control hives received pollen suspension without addition of probiotic lacto-bacilli. The suspensions were supplied to bees three times, once a week. Before and during administration of probiotics, samples of honey bees from each hive were taken every week and numbers of lactobacilli, P. larvae, enterobacteria and coliform bacteria were determined in their digestive tracts. Four weeks after the first administration of probiotic-pollen solution the numbers of lactobacilli were increased approximately by 0.5 log. Before starting this experiment, P. larvae (approx. 107 CFU.ml−1) were detected in digestive tracts of honey bees. In the experimental group after 3 weeks and in the control group after 4 weeks, no viable counts of P. larvae were found. The numbers of enterobacteria, coliform bacteria and Bacillus sp. decreased in both groups. During the experiment the health and condition of the hives were monitored. In addition, also monitored were: the development of the bee colony, the number of dead bees, the amount of hive debris, the aggressiveness of the bees, and the amount of honey spun; later during the autumn treatment also the fall of Varroa jacobsoni was noted. In the experimental and control groups, we recorded a decrease in the amount of hive debris. Only in the experimental group was a slightly above-average development of the bee colony, slightly above-average honey yields and a 70 % lower drop of Varroa mites. Based on these results, we assumed that the probiotic-pollen solution had a positive influence on the composition of microbiota in bee digestive tracts and it can increase resistance to P. larvae. It also had a positive effect on the health and condition of the bee colony. Probiotic-free pollen solution showed similar but weaker effects.

Alveolar echinococcosis (AE) caused by the larval stage of Echinococcus multilocularis is serious parasitic diseases associated with the host´s immunosuppression. The effects of human non-immune dialyzable leukocyte extract (DLE) on immune cells in blood and spleen and parasitic cysts weight in Balb/c mice after oral (PO), subcutaneous (SC) and intraperitoneal administration (IP) were compared. The reduction in cysts weight (p < 0.01) was recorded after PO route, whereas moderate reduction was found after SC and IP routes. The elevation of lymphoid populations in blood and spleen was found after PO administration (p < 0.01) in parallel with reduced myeloid population. Infection-elicited decline in B220+B cells was partially abolished by PO route, but DLE routes did not influence the CD3+ T cells. The proportions of CD3+CD4+Th lymphocytes were moderately upregulated, whereas CD3+CD8+Tc populations were reduced after all DLE routes (p < 0.01). PO administration increased CD11b+MHCIIhigh blood monocytes, CD11b-SigleF+ cell, but not CD11b+Si-glecF+ eosinophils in the blood, stimulated after SC and IP routes. DLE induced downregulation of NO production by LPS-stimulated adherent splenocytes ex vivo. Con A-triggered T lymphocyte proliferation was associated with the elevated IFN-γ production and transcription factor Tbet mRNA expression. The alleviation of Th2 (IL-4) and Treg (TGF-β) cytokine production by lymphocytes ex vivo paralleled with downregulation of gene transcription for cytokines, GATA and FoxP3. Reduction of myeloid cells with suppressive activity was found. The SC and IP routes affected partially the cysts weights, diminished significantly gene transcription, NO levels and Th2 and Treg cytokines production. Results showed that PO route of DLE administration was the most effective in ameliorating immunosuppression via stimulation of Th1 type, reducing Th2 and Treg type of immunity and CD3+CD8+Tc lymphocytes in the blood and spleens during E. multilocularis infection in mice.

This study aimed to evaluate the immunomodulatory effect of the probiotic Limosilactobacillus reuteri L26 Biocenol TM (L26) and its purified exopolysaccharide (EPS) with respect to antiviral innate immune response. In our experiment, we used porcine epithelial IPEC-J2 cells as a model of the intestinal barrier in a homologous infection by porcine Rotavirus   A  strain OSU6 (RVA). The production of selected molecules of non-specific humoral immunity was evaluated at the mRNA level. The EPS alone significantly increased the level of interferon λ3 (IFN-λ3) mRNA in the non-infected IPEC-J2 cells ( P  < 0.001). We also tested whether the treatment of IPEC-J2 cells by L26 or EPS influences the replication of RVA by virus titration and real-time PCR. We found that a pre-treatment in combination with subsequent continuous stimulation has no influence on the RVA replication. However, both treatments significantly decreased the RVA-induced production of IFN-λ3 ( P  < 0.05) and the "SOS" cytokine interleukin 6 (IL-6; P  < 0.01), already at the transcription level. In addition, the EPS treatment resulted in significantly increased IL-10 mRNA in the RVA-infected cells. In summary, we assume an immunoregulatory potential of  L. reuteri L26 Biocenol TM and its EPS in the local intestinal antiviral immune response., Competing Interests: The authors declare no conflict of interest., (Copyright: © 2024 Schusterova et al.)

Transplantation of faecal microbiota (FMT) is generally considered a safe therapeutic procedure with few adverse effects. The main factors that limit the spread of the use of FMT therapy for idiopathic inflammatory bowel disease (IBD) are the necessity of minimising the risk of infection and transfer of another disease. Obtaining the animal model of UC (ulcerative colitis) by exposure to DSS (dextran sodium sulphate) depends on many factors that significantly affect the result. Per os intake of DSS with water is individual for each animal and results in the development of a range of various forms of induced UC. For this reason, the aim of our study was to evaluate the modulation and regenerative effects of FMT on the clinical and histopathological responses and the changes in the bowel microenvironment in pseudo germ-free (PGF) mice of the BALB/c line subjected to chemical induction of mild, moderate and serious forms of UC. The goal was to obtain new data related to the safety and effectiveness of FMT that can contribute to its improved and optimised use. The animals with mild and moderate forms of UC subjected to FMT treatment exhibited lower severity of the disease and markedly lower damage to the colon, including reduced clinical and histological disease index and decreased inflammatory response of colon mucosa. However, FMT treatment failed to achieve the expected therapeutic effect in animals with the serious form of UC activity. The results of our study indicated a potential safety risk involving development of bacteraemia and also translocation of non-pathogenic representatives of bowel microbiota associated with FMT treatment of animals with a diagnosed serious form of UC.

Introduction: With growing significance in nervous system repair, mesenchymal stem cell-derived conditioned media (MSCCM) have been used in cell-free therapies in regenerative medicine. However, the immunomodulatory and neuroregenerative effects of MSCCM and the influence of priming on these effects are still poorly understood., Methods: In this study, by various methods focused on cell viability, proliferation, neuron-like differentiation, neurite outgrowth, cell migration and regrowth, we demonstrated that MSCCM derived from adipose tissue (AT-MSCCM) and amniotic membrane (AM-MSCCM) had different effects on SH-SY5Y cells., Results and Discussion: AT-MSCCM was found to have a higher proliferative capacity and the ability to impact neurite outgrowth during differentiation, while AM-MSCCM showed more pronounced immunomodulatory activity, migration, and re-growth of SH-SY5Y cells in the scratch model. Furthermore, priming of MSC with pro-inflammatory cytokine (IFN-γ) resulted in different proteomic profiles of conditioned media from both sources, which had the highest effect on SH-SY5Y proliferation and neurite outgrowth in terms of the length of neurites (pAT-MSCCM) compared to the control group (DMEM). Altogether, our results highlight the potential of primed and non-primed MSCCM as a therapeutic tool for neurodegenerative diseases, although some differences must be considered., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Hudakova, Mudronova, Marcincakova, Slovinska, Majerova, Maloveska, Petrouskova, Humenik and Cizkova.)

In this study, we provide comprehensive analyses of mesenchymal stem cells (MSCs) isolated from three types of canine tissues: bone marrow (BM-MSCs), adipose tissue (AT-MSCs) and amniotic tissue (AM-MSCs). We compare their morphology, phenotype, multilineage potential and proliferation activity. The BM-MSCs and AM-MSCs showed fibroblast-like shapes against the spindle shape of the AT-MSCs. All populations showed strong osteogenic and chondrogenic potential. However, we observed phenotypic differences. The BM-MSCs and AT-MSCs revealed high expression of CD29, CD44, CD90 and CD105 positivity compared to the AM-MSCs, which showed reduced expression of all the analysed CD markers. Similarly, the isolation yield and proliferation varied depending on the source. The highest isolation yield and proliferation were detected in the population of AT-MSCs, while the AM-MSCs showed a high yield of cells, but the lowest proliferation activity, in contrast to the BM-MSCs which had the lowest isolation yield. Thus, the present data provide assumptions for obtaining a homogeneous MSC derived from all three canine tissues for possible applications in veterinary regenerative medicine, while the origin of isolated MSCs must always be taken into account.

Due to the physiological complexity of the tumour, a single drug therapeutic strategy may not be sufficient for effective treatment. Emerging evidence suggests that combination strategies may be important to achieve more efficient tumour responses. Different immunomodulators are frequently tested to reverse the situation for the purpose of improving immune response and minimizing chemotherapy side effects. Immodin (IM) represents an attractive alternative to complement chemotherapy, which can be used to enhance the immune system after disturbances resulting from the side effects of chemotherapy. In the presented study, a model of CT26 tumor-bearing mice was used to investigate the effect of single IM or its combination with 5-fluorouracil (5-FU) on colon cancer cells. Our results highlight that the beneficial role of IM claimed in previous studies cannot be generalised to all chemotherapeutic drugs, as 5-FU toxicity was not increased. On the contrary, the chemotherapeutic anti-cancer efficacy of 5-FU was greatly compromised when combined with IM. Indeed, the combined treatment was significantly less effective regarding the tumour growth and animal survival, most probably due to the increased number of tumour-associated macrophages, and increased 5-FU cytotoxic effect related to kidneys and the liver.