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Lactobacillus helveticus Lafti L10 supplementation reduces respiratory infection duration in a cohort of elite athletes: a randomized, double-blind, placebo-controlled trial
No full textA randomized, double-blind, placebo-controlled study was conducted to evaluate if Lactobacillus helveticus Lafti L10 (Lallemand Health Solutions, Montreal, Que., Canada) supplementation during 14 weeks in winter can influence the duration, severity, and incidence of upper respiratory tract illness (URTI), as well as to monitor different immune parameters in the population of elite athletes. Before and after the treatment, cardiopulmonary testing and self-rated state of moods evaluation (by Profile of Mood States questionnaire) were performed and blood samples were collected. Thirty-nine elite athletes were randomized either to the placebo (n = 19) or the probiotic (n = 20) group. The probiotic group received L. helveticus Lafti L10, 2 x 10(10) Colony Forming Units. Lafti L10 significantly shortened the URTI episode duration (7.25 +/- 2.90 vs. 10.64 +/- 4.67 days, p = 0.047) and decreased the number of symptoms in the probiotic group (4.92 +/- 1.96 vs. 6.91 +/- 1.22, p = 0.035). Severity and incidence of URTI did not differ between the treatments. There were no significant changes in leukocyte subpopulation abundance, transforming growth factor-beta serum levels, level of interleukin-10 secreted from peptidoglican stimulated peripheral blood mononuclear cells (PBMCs), interferon-gamma level secreted from concanavalin A-stimulated PBMCs or viability/proliferation of PBMCs upon antigen stimulation. Group effect for CD4+/CD8+ ratio was significant (F-[1,F-37] = 6.99, p = 0.020, eta(2) = 0.350); this difference was not significant at baseline, but was evident after 14 weeks (p = 0.02). A significant interaction effect was noted for self-rated sense of vigor (F-[1,F-37] = 11.76, p = 0.009, eta(2) = 0.595). Self-rated sense of vigor increased in the probiotic group (18.5 +/- 4.1 vs. 21.0 +/- 2.6, p = 0.012). Probiotic strain Lafti L10 can be a beneficial nutritional supplement for the reduction of URTI length in elite athletes.Peer-reviewed manuscript: [http://intor.torlakinstitut.com/handle/123456789/607
Gut-associated lymphoid tissue, gut microbes and susceptibility to experimental autoimmune encephalomyelitis
No full textGut microbiota and gut-associated lymphoid tissue have been increasingly appreciated as important players in pathogenesis of various autoimmune diseases, including multiple sclerosis. Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis that can be induced with an injection of spinal cord homogenate emulsified in complete Freund's adjuvant in Dark Agouti (DA) rats, but not in Albino Oxford (AO) rats. In this study, mesenteric lymph nodes (MLN), Peyer's patches (PP) and gut microbiota were analysed in these two rat strains. There was higher proportion of CD4(+) T cells and regulatory T cells in non-immunised DA rats in comparison to AO rats. Also, DA rat MLN and PP cells were higher producers of pro-inflammatory cytokines interferon-gamma and interleukin-17. Finally, microbial analyses showed that uncultivated species of Turicibacter and Atopostipes genus were exclusively present in AO rats, in faeces and intestinal tissue, respectively. Thus, it is clear that in comparison of an EAE-susceptible with an EAE-resistant strain of rats, various discrepancies at the level of gut associated lymphoid tissue, as well as at the level of gut microbiota can be observed. Future studies should determine if the differences have functional significance for EAE pathogenesis
Vrste roda Trichoderma, uzročnici zelene plesni šampinjona [Agaricus bisporus (Lange) Imbach] i njihova osetljivost na fungicide i biofungicide
No full textTrichoderma species, the causal agents of green mould disease, induce great losses in button mushroom (Agaricus bisporus) farms. Twenty Trichoderma isolates were collected on 13 Serbian A. bisporus farms and one in Bosnia and Herzegovina during 2006 – 2010. Twelve isolates were classified into five species by standard mycological studies and ITS1/ITS4 sequence analyses, namely Trichoderma atroviride, Trichoderma koningii, Trichoderma virens, Trichoderma aggressivum f. europaeum and Trichoderma harzianum. Eight isolates were not identified to the species level but were shown to be related to T. harzianum. The isolates of T. harzianum exhibited the highest virulence to the harvested A. bisporus pilei and T. virens and T. aggressivum f. europaeum the lowest. Fungicides are widely used to control mushroom diseases although green mould control is encumbered with difficulties. The aims of this study were therefore to research in vitro toxicity of several commercial fungicides to Trichoderma isolates originating from Serbian and Bosnia-Herzegovina farms, and to evaluate the effects of pH and light on their growth. The majority of isolates demonstrated optimal growth at pH 5.0, and the rest at pH 6.0. A few isolates also grew well at pH 7. The weakest mycelial growth was noted at pH 8.0 – 9.0. Generally, light had an inhibitory effect on the growth of tested isolates. The isolates showed the highest susceptibility to prochloraz-Mn, chlorothalonil and carbendazim (ED50 less than 1 mg/L), and were less sensitive to iprodione (ED50 ranged 0.84 – 6.72 mg/L), weakly resistant to thiophanate-methyl (ED50 = 3.75 – 24.13 mg/L), and resistant to trifloxystrobin (ED50 = 10.25 – 178.23 mg/L). Considering the toxicity of fungicides to A. bisporus, prochloraz-Mn and carbendazim showed the best selective toxicity (0.05 and 0.02 respectively), iprodione and chlorothalonil moderate (0.16), and thiophanate-methyl the lowest (1.24), while trifloxystrobin toxicity to A. bisporus was not tested because of its inefficiency against Trichoderma isolates. The best selective toxicity to both pathogen and the host showed prochloraz-Mn and carbendazim with values less than 0.1. Carbendazim was withdrawn from the market and therefore prochloraz-Mn could be recommended for mushroom cultivation in Serbia. Antifungal activity of two biofungicides based on Bacillus subtilis and tea tree oil were tested in vitro to all Trichoderma isolates. B. subtilis was highly toxic to all tested Trichoderma isolates, their ED50 values were below 1.3 mg/L. Tea tree oil did not exhibit a significant antifungal activity (ED50 = 11.9 – 370.8 mg/L). The efficacy of biofungicides was evaluated against T. harzianum in a mushroom growing room, and they were applied alone or in combination with the fungicide at a respective proportion of 20:80%. Prochloraz-Mn showed higher efficacy than both tested biofungicides or their respective mixtures. The biofungicide based on B. subtilis demonstrated greater efficacy in preventing disease symptoms than tea tree oil. B. subtilis combined with the fungicide revealed less antagonism in efficacy against pathogen than tea tree oil.Pojedine vrste roda Trichoderma su uzročnici zelene plesni šampinjona (Agaricus bisporus) i nanose velike materijalne štete u gajilištima. Od 2006. do 2010. godine prikupljeno je 20 izolata roda Trichoderma iz 13 gajilišta u Srbiji i jednog iz Bosne i Hercegovine. Dvanaest izolata je klasifikovano na osnovu standardnih mikoloških metoda i analizom ITS1/ITS4 sekvence u pet vrsta: Trichoderma atroviride, Trichoderma koningii, Trichoderma virens, Trichoderma aggressivum f. europaeum i Trichoderma harzianum. Osam izolata nije identifikovano do nivoa vrste ali je pokazano da su filogenetski bliski sa vrstom T. harzianum. Najvišu virulentnost na ubranom šeširu A. bisporus je pokazala vrsta T. harzianum, a najnižu T. virens i T. aggressivum f. europaeum. Većina izolata je najbolje rasla na podlozi koja je imala pH = 5, potom na pH = 6, par izolata na pH = 7, dok je najslabiji rast zabeležen na pH = 8 – 9. Generalno, svetlost je imala inhibitorni efekat na rast izolata. Fungicidi su u širokoj upotrebi u kontroli bolesti šampinjona, iako je kontrola zelene plesni otežana. Stoga, jedan od ciljeva istraživanja je bio da se ispita in vitro toksičnost nekoliko dostupnih komercijalnih fungicida na prikupljene izolate roda Trichoderma. Testirani izolati su bili najosetljivi na prohloraz-Mn, hlorotalonil i karbendazim (EC50 1 mg/L), osetljivi na iprodion (EC50 = 0,84 – 6,72 mg/L), umereno rezistentni na tiofanat-metil (EC50 = 3,75 – 24,13 mg/L) i rezistentni na trifloksistrobin (EC50 = 10,25 – 178,23 mg/L). Proučavajući toksičnost fungicida na A. bisporus, najbolju selektivnu toksičnost su pokazali prohloraz-Mn (0,05) i karbendazim (0,02), iprodion i hlorotalonil umerenu (0,16), tiofanat-metil najnižu (1,24), dok toksičnost trifloksistrobina za A. bisporus nije testirana zbog nezadovoljavajuće toksičnosti za Trichoderma spp. izolate. Najbolju selektivnu toksičnost za patogenu gljivu i gljivu domaćina su pokazali prohloraz-Mn i karbendazim sa vrednostima nižim od 0,1. Pošto se karbendazim povlači sa tržišta, prohloraz-Mn je fungicid koji se može preporučiti za primenu u gajilištima za suzbijanje zelene plesni. Antifungalna aktivnost dva biofungicida, na bazi Bacillus subtilis i ulja čajnog drveta, je testirana in vitro na sve Trichoderma spp. izolate. B. subtilis je bio visoko toksičan za sve testirane izolate (EC50 lt 1,3 mg/L), dok ulje čajnog drveta nije pokazalo značajnu antifungalnu aktivnost (EC50 = 11,9 – 370,8 mg/L). Efikasnost biofungicida protiv vrste T. harzianum je ocenjena u oglednom gajilištu. Biofungicidi su primenjeni zasebno ili u kombinaciji sa fungicidom, prohloraz- Mn, u proporciji 20:80%. Prohloraz-Mn se pokazao efikasnijim od oba biofungicida kao i od kombinovane primene biofungicida i fungicida. Biofungicid na bazi B. subtilis je bio efikasniji od ulja čajnog drveta u pogledu sprečavanja pojave simptoma bolesti. Takođe, B. subtilis je pokazao manji antagonistički efekat u efikasnosti protiv patogena od ulja čajnog drveta kada se primenjuju u kombinaciji sa prohloraz-Mn fungicidom
Effects of orally applied Fes p1-displaying L. plantarum WCFS1 on Fes p1 induced allergy in mice
Get PDFGroup I grass pollen allergens are major contributors to grass pollen-related seasonal allergic rhinitis, and as such a primary target for allergen specific immunotherapy. In this study the potential therapeutic role of oral application of Lactobacillus plantarum WCFS1, directing cell wall attachment of the recombinant Fes p1 allergen, from Festuca pratensis was tested in a mouse model of Fes p1 allergy. For surface expression of Fes p1 allergen in L. plantarum WCFS1 pSIP system with inducible expression was used. Balb/c mice were sensitized with Fes p1 protein in alum and subsequently received live recombinant L. plantarum orally. Antibody levels (IgE, total IgG, IgG1, IgG2a, and IgA) were determined by ELISA. Differential eosinophil count in peripheral blood was performed. Reduced peripheral blood eosinophilia and increased serum IgG2A levels was detected in both groups which received live L. plantarum orally. Specific serum IgA levels were increased only in mice treated with the recombinant bacteria. Oral application of L. plantarum WCFS1 has a beneficial therapeutic effect in a mouse model of Fes p1 allergy. Cell surface expression of Fes p1 allergen potentiates this phenomenon in an allergen specific way. (C) 2015 Elsevier B.V. All rights reserved.This is the peer‐reviewed version of the article: Minić, R.; Gavrović-Jankulović, M.; Petrušić, V.; Živković, I.; Eijsink, V. G. H.; Dimitrijević, L.; Mathiesen, G. Effects of Orally Applied Fes P1-Displaying L. Plantarum WCFS1 on Fes P1 Induced Allergy in Mice. Journal of Biotechnology 2015, 199, 23–28. [https://doi.org/10.1016/j.jbiotec.2015.01.028]
Capability of exopolysaccharide-producing Lactobacillus paraplantarum BGCG11 and its non-producing isogenic strain NB1, to counteract the effect of enteropathogens upon the epithelial cell line HT29-MTX
No full textThe putative protective role of the exopolysaccharide (EPS)-producing Lactobacillus paraplantarum BGCG11, and its non-EPS-producing isogenic strain NB1, was tested upon HT29-MTX monolayers challenged with seven opportunistic pathogens. The probiotic strain Lactobacillus rhamnosus LMG18243 (GG) was used as a reference bacterium. Tested lactobacilli were able to efficiently reduce the attachment to HT29-MTX of most pathogens. Lb. paraplantarum NB1 and Lb. rhamnosus GG were more efficient reducing the adhesion of Clostridium difficile or Yersinia enterocolitica than Lb. paraplantarum BGCG11, while stain BGCG11 reduced, to a greater extent, the adhesion of Escherichia coli and Listeria monocytogenes. The detachment and cell lysis of HT29-MTX monolayers in the presence of pathogens alone and co-incubated with lactobacilli or purified EPS was followed. L. monocytogenes induced the strongest cell detachment among the seven tested pathogens and this effect was prevented by addition of purified EPS-CG11. The results suggest that this EPS could be an effective macromolecule in protection of HT29-MTX cells from the pathogen-induced lysis. Regarding innate intestinal barrier, the presence of C. difficile induced the highest IL-8 production in HT29-MTX cells and this capability was reinforced by the co-incubation with Lb. paraplantarum NB1 and Lb. rhamnosus GG. However, the increase in IL-8 production was not noticed when C difficile was co-incubated with EPS-producing Lb. paraplantarum BGCG11 strain or its purified EPS-CG11 polymer, thus indicating that the polymer could hinder the contact of bacteria with the intestinal epithelium. The measurement of mucus secreted by HT29-MTX and the expression of mud, muc2, muc3B and muc5AC genes in the presence of pathogens and lactobacilli suggested that all lactobacilli strains are weak "co-adjuvants" helping some pathogens to slightly increase the secretion of mucus by HT29-MTX, while purified EPS-CG11 did not induce mucus secretion. Taking altogether, Lb. paraplantarum BGCG11 could act towards the reinforcement of the innate mucosal barrier through the synthesis of a physical-protective EPS layer which could make difficult the contact of the pathogens with the epithelial cells.Peer-reviewed manuscript: [https://intor.torlakinstitut.com/handle/123456789/708
Characterization of lactic acid bacteria isolated from Bosnian artisanal dry fermented sausage sudzuk) during fermentation
Get PDFBosnian sudzuk is a dry fermented sausage produced in a rural household near the town of Visoko in central Bosnia and Herzegovina. This kind of sausage was manufactured only from beef and spices in a traditional way without the addition of a starter cultures. To identify lactic acid bacteria (LAB), a total number of 160 LAB strains were isolated from five samples of Bosnian sudzuk collected over 28 days of fermentation. Preliminary identification by phenotypic tests and 16S rDNA sequencing were performed for all 160 of the LAB isolates. Identification of LAB strains from traditionally produced Bosnian sausage at the species level revealed the presence of six genera: Lactococcus sp., Enterococcus sp., Leuconostoc sp., Lactobacillus sp., Pediococcus sp. and Weissella sp.. Among the 15 distinct species identified, the species Lactobacillus plantarum, Leuconostoc mesenteroides, Lactococcus lactis, Enterococcus faecalis and Enterococcus durans were present throughout the entire process of fermentation. Leuconostoc mesenteroides, Lactobacillus plantarum and Lactococcus lactis prevailed, with 21.8%, 19.3% and 13.1%, respectively, of total LAB strains during the entire fermentation process. Significant negative correlations (r = 0.892 and r = 0.829, respectively) between the presence of Weissella sp. and Lactobacillus sp., and between the presence of Weissella sp. and Lactococcus sp. were recorded. Lactobacillus plantarum, Enterococcus durans and Leuconostoc mesenteroides were the best producers of aromogenic compounds while 32.3% of Lactobacillus plantarum and 28.6% of Leuconostoc mesenteroides were produced exopolysaccharides
Modulation of macrophage reactive oxygen and nitrogen species production by neuropeptide Y
No full textNeuropeptide Y (NPY), containing 36 amino acids, is one of the most evolutionary
conserved peptides. Due to a very wide tissue distribution of NPY and its significance for the
human and animal physiology, peptides with high structural homology to NPY, e.g., peptide
YY (PYY), pancreatic polypeptide (PP) and their truncated forms NPY2-36, NPY3-36 and
PY Y3-36, are specified as members of NPY family. Beside effects on the central nervous
system, including regulation of food intake, energy balance, nociception, anxiety and sexual
behavior, NPY in the periphery has a potent mitogenic activity, stimulates angiogenesis and 15
chemotactic for vascular smooth muscle cells. NPY-related peptides are potent inhibitors of
intestinal fluid secretion and motility. However, the presence of NPY in the cells of the
immune system, together with the expression of specific Y-receptor subtypes (Y1, Y2 and
Y5) on immune cells, suggested ability of autocrine/paracrine immune regulation by
endogenous NPY and related peptides. Experimental evidences revealed that NPY in vitro
stimulated rat macrophage reactive oxygen species production ма Y] and Y2 receptors,
whereas activation of Y5 receptors suppressed oxidative burst in these cells. Quite the
opposite, NPY in vitro considerably decreased rat granulocyte peroxide production, and this
effect was mediated by Y2 and Y5 receptors. Regarding the production of NO by LPSactivated
macrophages, NPY-induced augmentation involved YI and Y2, and excluded
mediation via Y5 receptors. Furthermore, NPY enhanced NO production in macrophages
from young rats, whereas it was ineffective in this respect in macrophages from aged rats. As
a whole, direct effect of NPY on reactive oxygen and nitrogen species production is
dependent on cell type, engagement of specific NPY receptors and age
Capability of exopolysaccharide-producing Lactobacillus paraplantarum BGCG11 and its non-producing isogenic strain NB1, to counteract the effect of enteropathogens upon the epithelial cell line HT29-MTX
Get PDFThe putative protective role of the exopolysaccharide (EPS)-producing Lactobacillus paraplantarum BGCG11, and its non-EPS-producing isogenic strain NB1, was tested upon HT29-MTX monolayers challenged with seven opportunistic pathogens. The probiotic strain Lactobacillus rhamnosus LMG18243 (GG) was used as a reference bacterium. Tested lactobacilli were able to efficiently reduce the attachment to HT29-MTX of most pathogens. Lb. paraplantarum NB1 and Lb. rhamnosus GG were more efficient reducing the adhesion of Clostridium difficile or Yersinia enterocolitica than Lb. paraplantarum BGCG11, while stain BGCG11 reduced, to a greater extent, the adhesion of Escherichia coli and Listeria monocytogenes. The detachment and cell lysis of HT29-MTX monolayers in the presence of pathogens alone and co-incubated with lactobacilli or purified EPS was followed. L. monocytogenes induced the strongest cell detachment among the seven tested pathogens and this effect was prevented by addition of purified EPS-CG11. The results suggest that this EPS could be an effective macromolecule in protection of HT29-MTX cells from the pathogen-induced lysis. Regarding innate intestinal barrier, the presence of C. difficile induced the highest IL-8 production in HT29-MTX cells and this capability was reinforced by the co-incubation with Lb. paraplantarum NB1 and Lb. rhamnosus GG. However, the increase in IL-8 production was not noticed when C difficile was co-incubated with EPS-producing Lb. paraplantarum BGCG11 strain or its purified EPS-CG11 polymer, thus indicating that the polymer could hinder the contact of bacteria with the intestinal epithelium. The measurement of mucus secreted by HT29-MTX and the expression of mud, muc2, muc3B and muc5AC genes in the presence of pathogens and lactobacilli suggested that all lactobacilli strains are weak "co-adjuvants" helping some pathogens to slightly increase the secretion of mucus by HT29-MTX, while purified EPS-CG11 did not induce mucus secretion. Taking altogether, Lb. paraplantarum BGCG11 could act towards the reinforcement of the innate mucosal barrier through the synthesis of a physical-protective EPS layer which could make difficult the contact of the pathogens with the epithelial cells.This is the peer reviewed version of the paper: Zivkovic, M., Hidalgo-Cantabrana, C., Kojic, M., Gueimonde, M., Golic, N., & Ruas-Madiedo, P. (2015). Capability of exopolysaccharide-producing Lactobacillus paraplantarum BGCG11 and its non-producing isogenic strain NB1, to counteract the effect of enteropathogens upon the epithelial cell line HT29-MTX. Food Research International, 74, 199–207.[ https://doi.org/10.1016/j.foodres.2015.05.012]Published version : [https://intor.torlakinstitut.com/handle/123456789/707
Key protection factors against tetanus: Anti-tetanus toxin antibody affinity and its ability to prevent tetanus toxin - ganglioside interaction
No full textAntibodies capable to neutralize tetanus toxin (TeNT) are key factors in protection against tetanus disease. Although antibody-based therapeutics for treatment of tetanus exist on the market its production is tedious. Hence, the tetanus-specific antibodies preparation that could be easily produced in large scale in vitro would be beneficial. Monoclonal antibodies (MAbs) are considered for a long time as a reagent of choice, but the core drawback is how to select a MAb that would be safe in providing efficacious protection. In this study we have investigated the parameters crucial for a single MAb to be assigned as protective. Eight murine MAbs were characterized in vitro for their reactivity toward TeNT and assessed in vivo for protectiveness against TeNT intoxication. Correlation of in vitro and in vivo data has revealed that in vitro selection of MAb that is protective in vivo could be performed by a combination of two assays: the measurement of MAb affinity toward TeNT taking Ka 1 x 10(8) M-1 as a threshold level, and the evaluation of its capability to prevent TeNT-ganglioside interaction. Single MAb could be taken into consideration as a potential therapeutic only if it has a capacity to completely inhibits TeNT-ganglioside complex formation. (C) 2015 Elsevier Ltd. All rights reserved
Composition of polyphenol and polyamide compounds in common ragweed (Ambrosia artemisiifolia L.) pollen and sub-pollen particles
No full textPhenolic composition of Ambrosia artemisiifolia L. pollen and sub-pollen particles (SPP) aqueous extracts was determined, using a novel extraction procedure. Total phenolic and flavonoid content was determined, as well as the antioxidative properties of the extract. Main components of water-soluble pollen phenolics are monoglycosides and malonyl-mono- and diglycosides of isorhamnetin, quercetin and kaempferol, while spermidine derivatives were identified as the dominant polyamides. SPP are similar in composition to pollen phenolics (predominant isorhamnetin and quercetin monoglycosides), but lacking small phenolic molecules ( lt 450 Da). Ethanol-based extraction protocol revealed one-third lower amount of total phenolics in SPP than in pollen. For the first time in any pollen species, SPP and pollen phenolic compositions were compared in detail, with an UHPLC/ESI-LTQ-Orbitrap-MS-MS approach, revealing the presence of spermidine derivatives in both SPP and pollen, not previously reported in Ambrosia species. (C) 2014 Elsevier Ltd. All rights reserved