Improving Health Through Evidence-Based Probiotics

Improving health through evidence-based probiotics

Company Profile & Probiotic Portfolio

wincloveprobiotics.com vw

Company Profile

Winclove Probiotics

Developing probiotic formulations since 1991 with a strong emphasis on research

Winclove Probiotics, established in Amsterdam, The Netherlands, has been specializing in research, development and manufacturing of probiotic formulations for over 25 years. Our probiotic formulations are developed in close collaboration with leading research institutes, universities and academic hospitals. This enables us to develop innovative multispecies probiotic formulations for many different health indications. We continuously invest in research and technologies in order to develop the most effective probiotics that ensure end-users with the best solution. Looking for business opportunities?

Winclove’s mission is to create long-lasting, sustainable and strategic partnerships. To achieve this, we offer innovative, high-quality and competitive probiotic solutions, as well as in-house scientific expertise and sales support. Our indication-specific formulations are designed for specific microbiota-related indications. The thorough substantiation of our probiotic formulations with scientific and clinical evidence makes them ideal for selling medically endorsed. We are a committed business partner and are looking forward to starting a fruitful collaboration with you!

Product package

Dossiers Technical lab analysis, in vitro data, etc. Clinical in vivo studies, Development of Probiotic blend post market studies, Research the formulation user trials collaboration

• Training and education for salesforce & healthcare professionals • Round tables with KOLs • Sharing best practices other countries Training • Monthly news update Sales & marketing QA & RA & education • Independent scientific conferences support Probiotic portfolio

Ecologic® Reducing vulnerability Brain formulations to depression Our Ecologic® formulations have Improving brain functioning under stress been developed in collaboration with leading universities and academic hospitals worldwide. For Managing allergic Allergy every indication bacterial strains are symptoms; hay fever selected based on characteristics and and eczema functionality, and the formulation has Preventing early-onset been proven effective in clinical trials. eczema

Winclove formulations Reducing antibiotic- Gut associated side effects Winclove formulations consist of carefully selected probiotic strains Reducing constipation and are based on scientific and in Reducing exercise- vitro evidence. induced stress

Key strengths of our Preventing traveler’s diarrhea formulations • Indication-specific selection of bacteria Improving intestinal Age • Evidence-based formulations health in newborns • Stability at room temperature, Improving intestinal no refrigeration needed health in children • Excellent survival of the Improving intestinal gastro-intestinal tract health in adults • High viability and metabolically active bacteria Improving intestinal health of elderly • Flexible packaging; from bulk delivery to fully packed

PROBIOACT® TECHNOLOGY

Protective and nutritional ingredients, specifically selected and tested to increase: • Shelf life of the formulation • Gastro-intestinal survival of the bacteria • Activity of the bacteria in the intestine Research and development areas

Gut-brain

Ear, nose & throat

Allergy

Gastrointestinal

Drug related effects

Cardiometabolic

Urogenital

PIONEER IN PROBIOTIC RESEARCH

Over the last decades, driven by the fascination for bacteria, Winclove has built an extensive database of well characterized probiotic strains. This unique knowledge enables us to develop evidence-based multispecies probiotic formulations for a wide range of microbiota-related indications. In collaboration with research partners, these formulations are tested in clinical trials and published in peer-reviewed journals.

This information is for professionals only and should not be given to consumers - Reducing vulnerability to depression - Improving brain functioning under stress

Strain selection Bacterial strains

The bacterial strains in Ecologic® BARRIER have been selected for Ecologic® BARRIER is a multispecies formulation consisting of 9 bac- their capacity to strengthen the intestinal barrier function and reduce terial strains, 2,5 x 109 CFU/gram. The recommended dosage is 2 low grade inflammation1. The strains have been screened in vitro for grams a day. their capacity to: • B. bifidum W23 • L. acidophilus W37 • L. salivarius W24 • improve the intestinal barrier function • B. lactis W51 • L. brevis W63 • Lc. lactis W19 • inhibit mast cell activation • B. lactis W52 • L. casei W56 • Lc. lactis W58 • stimulate IL-10 production • break down lipopolysaccharides (LPS).

Clinical evidence The effect of Ecologic® BARRIER on vul- Multiple, placebo-controlled, rat studies Moreover, researchers from Radboud Uni- nerability for depression was tested in a were performed with Ecologic® BARRI- versity Nijmegen and Donders Intsitute for triple-blind, placebo-controlled, rand- ER by Aarhus University, Denmark, using Brain Cognition and Behavior, The Nether- omized, human trial, performed by Leiden wild type rats and a genetic rat model lands, performed a double-blind, placebo- University, The Netherlands. The research- for depression.3-6 In the genetic model, controlled, randomized, human trial with ers investigated the effect of Ecologic® Ecologic® BARRIER reduced risk taking be- Ecologic® BARRIER. The researchers BARRIER on cognitive reactivity to sad havior and prevented the exacerbation of investigated the effect on neurocog- mood using the Leiden Index of depres- depressive-like behavior normally induced nition, using functional magnetic res- sion sensitivity scale (LEIDS-r). 40 healthy by a high-fat diet. In the wild-type rats, onance imaging (fMRI), and stress-in- participants received 2 grams Ecologic® Ecologic® BARRIER significantly reduced duced working memory performance BARRIER or placebo for 4 weeks. The re- depressive-like behavior, measured by meausured by a Digit Span Backwards sults showed that Ecologic® BARRIER sig- a forced swim test (see figure 2), and dif- test. 58 participants received 2 grams of nificantly reduced overall cognitive reac- ferences in gut microbiota between re- Ecologic® BARRIER or placebo for 4 tivity to sad mood2 (see figure 1). sponder and non-responder animals were weeks. Ecologic® BARRIER significant- identified. Ecologic® BARRIER influenced ly increased working memory perfor- gene-expression in the hippocampus mance7 (see figure 3). Ecologic® BARRIER Figure 1: LEIDS-r scores before and after 4 weeks of supplementation with Ecologic® BARRIER. and altered immunological and metabolic buffered against the damaging effects of *Significant decrease, p<0.001. markers, explaining the positive outcomes. stress on brain functioning.

Figure 2: Seconds of immobility during the Forced Figure 3: Stress-induced changes in working memory. Swim Test. *Significant decrease, p<0.001. Calculated as the difference of DS backwards scores after stress minus scores before stress. *Significant differences, p<0.05. LEIDS-r scores

Immobility (sec)

DS backwards, stress after-before Baseline Intervention Placebo Ecologic® BARRIER Placebo pre- test Probiotics pre- test Placebo Ecologic® BARRIER Placebo post- test Probiotics post- test References Ecologic® BARRIER publications 1. van Hemert et al. Influence of the multispecies probiotic Ecologic® BARRIER on parame- 5. Tillmann, et al. Probiotics reduce risk-taking behavior in the Elevated Plus Maze in the ters of intestinal barrier function. Food and Nutrition Sciences 2014. Flinders Sensitive Line rat model of depression. Behav Brain Res 2019;359:755-762. 2. Steenbergen et al. A randomized controlled trial to test the effect of multispecies probi- 6. Abildgaard, et al. The antidepressant-like effect of probiotics and their faecal abundance otics on cognitive reactivity to sad mood. Brain Behav Immun 2015;48:258-64. may be modulated by the cohabiting gut microbiota in rats. Eur Neuropsychopharmacol 3. Abildgaard, et al. Probiotic treatment reduces depressive-like behaviour in rats inde- 2019;29(1):98-110. pendently of diet. Psychoneuroendocrin 2017;79:40-48. 7. Papalini, et al. Stress matters: Randomized controlled trial on the effect of probiotics on 4. Abildgaard, et al. Probiotic treatment protects against the pro-depressant-like effect of neurocognition, Neurobiology of Stress 2019;10:100141 high-fat diet in Flinders Sensitive Line rats. Brain Behav Immunity 2017;65:33–42. Managing allergic symptoms: hay fever and eczema

Strain selection Bacterial strains

The probiotic strains in Ecologic® ALLERGYCARE have been selected Ecologic® ALLERGYCARE is a multispecies formulation consisting of to influence the immune system. The strains have been screened in 6 probiotic strains, 1 x 109 CFU/gram. The recommended dosage is 2 vitro for their capacity to: grams, twice daily. • Modulate production of immunosuppressive cytokines by; • B. bifidum W23 • L. salivarius W57 - induction of IL-10 and IFN-y • B. lactis W51 • Lc. lactis W58 - reduction of IL-4, IL-5 and IL-13. • L. acidophilus W55 vitamin B2 and biotin • L. casei W56

Clinical evidence The effect of Ecologic® ALLERGYCARE RAD) index. (see figure 1). These results was determined using the rhinitis qual- on atopic dermatitis (AD) symptoms are supported by the finding that total ity of life questionnaire (RQLQ). During has been tested in a double-blind, IgE levels were significantly dropped in the intervention patients also record- randomized, placebo-controlled human the probiotic group, compared to the ed AR symptoms and medication use. trial, performed by Harran University, placebo group. Ecologic® ALLERGYCARE significantly Turkey1. Forty children between 1-13 years improved QoL in 63% of the patients of age suffering from AD received 2x109 The effect of Ecologic® ALLERGYCARE (meaningful improvement >0.7 decrease CFU Ecologic® ALLERGYCARE or pla- on quality of life (QoL) in patients with in the RQLQ, p=0.005). The average cebo twice daily for eight weeks. Both allergic rhinitis (AR) has been tested in a RQLQ score dropped from 2.83 at base- clinical and mechanistic effects were clinical trial performed by Griffith Univer- line to 1.66 at midpoint to 1.38 at week 8 measured at baseline and at the end sity Australia2,3. Forty hay fever patients (see figure 2). In line with these findings, a of the intervention period. Ecologic® received 2x109 CFU Ecologic® ALLERGY- trend towards reduction in AR symptoms ALLERGYCARE significantly reduced CARE twice daily for eight weeks. Before and medication use was found. the SCORing Atopic Dermatitis (SCO- and after the intervention period, QoL

Figure 1: Changes in the SCORAD-index before and after the intervention. Figure 2: RQLQ scores during the intervention. * Significant decrease, p=0.005. * Significant decrease, p=0.005.

* * SCORAD % RQLQ score RQLQ * * * *

Placebo Baseline 8 weeks Ecologic® ALLERGYCARE Baseline 4 weeks 8 weeks supplementation supplementation supplementation

References Ecologic® ALLERGYCARE publications

1. Yesilova Y, Calka O, et al. Effect of probiotics on the treatment of children with atopic 3. Watts AM, Cox AJ, et al. A Specifically Designed Multispecies Probiotic Supplement dermatitis. Ann Dermatol 2012;24:189-93. Relieves Seasonal Allergic Rhinitis Symptoms. Altern Complement Med. 2018;24(8):833- 2. Watts AM, et al. Probiotics and Allergic Rhinitis: A Simon Two-Stage Design to Determine 840. Effectiveness. J Altern Complement Med. 2016;22(12):1007-1012.

Preventing early onset eczema

Strain selection Bacterial strains

The probiotic strains in Ecologic® PANDA have been selected for Ecologic® PANDA is a multispecies formulation consisting of 4 probi- their capacity to strengthen the intestinal barrier function and influ- otic strains, 1 x 109 CFU/gram. The recommended dosage is 3 grams ence the immune system1,2. The strains have been screened in vitro a day. The treatment period for pregnant women is 6-8 weeks for their capacity to: before delivery and for babies the first year of life. • improve the intestinal barrier function • B. bifidum W23 • B. lactis W52 • modulate the production of immunosuppressive cytokines. • B. lactis W51 • Lc. lactis W58

Clinical evidence

Ecologic® PANDA has been tested in a years after birth3 (figure 1). Results sub- indicating good GI-survival of the bac- randomized, double-blinded, place- stantiated previous research outcomes terial strains6. Also, a possible mecha- bo-controlled trial including 102 pregnant by showing that babies who did not de- nism for the explanation of the positive women who were at high risk of getting velop eczema had a higher microbial di- outcomes of the PandA trial was found. an allergic baby due to a positive family versity. All phases of the research that led Higher concentrations of SCFAs were history (the PandA study). Subjects re- to this formulation have been published found in faecal samples of the children ceived either Ecologic® PANDA or a pla- in a PhD thesis4. who received Ecologic® PANDA com- cebo, which was administered prenatally pared to the placebo group7 (figure 2). (in the diet during pregnancy 6-8 weeks The children in the PandA trial were fol- The findings are in line with other stud- prior to delivery) and postnatally (during lowed till the age of 5 years. No long- ies that have shown the potential role of the first 12 months of life to the child). term effects were found,5 which is in SCFAs in eczema. The results showed a significant reduc- line with similar other studies. The re- tion of eczema after 3 months in the searchers did find higher quantities of In an observational trial it has been found babies receiving Ecologic® PANDA, and Ecologic® PANDA strains in microbiota that Ecologic® PANDA also benefits chil- a sustainment of this effect until two samples during the intervention period, dren after birth by reducing colic8.

Figure 1: Ecologic® PANDA shows a significant reduction of eczema Figure 2: Faecal concentrations SCFAs by 3 months of age. development.* Significant effect, p<0.05.

3 12 24 Months Acetate Propionate n-Butyrate Total SCFA CUMULATIVE INCIDENCE OF ATOPIC ECZEMA INCIDENCE OF ATOPIC CUMULATIVE Placebo Ecologic® PANDA Placebo Ecologic® PANDA

References Ecologic® PANDA publications

1. Niers L.E. et al. Identification of strong interleukin-10 inducing lactic acid bacteria which down-regulate T 5. Gorissen et al. Preventive effects of selected probiotic strains on the development of asthma and allergic helper type 2 cytokines. Clin Exp Allergy. 2005;35(11):1481-1489. rhinitis in childhood. The Panda study. Clin Exp Allergy. 2014;44(11):1431-3. 2. Niers L.E. et al. Selection of probiotic bacteria for prevention of allergic diseases: immunomodulation of 6. Rutten N.B.M.M. et al. Long Term Development of Gut Microbiota Composition in Atopic Children: Impact neonatal dendritic cells. Clin Exp Immunol. 2007;149(2):344-352. of Probiotics. PLoS ONE. 2015;10(9): e01376812015. 3. Niers L.E. et al. The effects of selected probiotic strains on the development of eczema (the PandA study). 7. Kim H.K. et al. Probiotic supplementation influences faecal short chain fatty acids in infants at high risk for Allergy. 2009;64(9):1349-1358. eczema. Benef Microbes. 2015; 6(6): 783-790. 4. Niers L.E. Probiotic bacteria for prevention of atopic disease. Design and application. PhD thesis. November 8. Hofmann H. Probiotics for indigestion in pregnancy and infant colic. Gyneacology, 26-11-2015. 2009. Reducing antibiotic-associated side effects

Strain selection Bacterial strains

The probiotic strains in Ecologic® AAD have been selected for their Ecologic® AAD is a multispecies formulation consisting of 9 probiot- capacity to inhibit antibiotic-associated diarrhoea (AAD)-related ic strains, 1 x 109 CFU/gram. The recommended dosage is 5 grams, pathogens. The strains have been screened in vitro for their capacity twice daily. It is adviced to start supplementation from the first day to inhibit growth of the pathogens: of antibiotic treatment until one week after cessation. • Clostridium difficile • B. bifidum W23 • L. paracasei W20 • Clostridium perfringens • B. lactis W51 • L. plantarum W62 • Entercoccus faecalis • E. faecium W54 • L. rhamnosus W71 • Escheria coli • L. acidophilus W37 • L. salivarius W24 • Bacillus subtilus. • L. acidophilus W55

Clinical evidence

Ecologic® AAD has been tested in a rand- one of the important mechanisms deter- 5-39%. omized, double-blind, placebo-controlled mining the efficacy of probiotics in AAD Furthermore, in a retrospective case re- trial with healthy volunteers taking amox- (figure 2). Ecologic® AAD was also able to port series of 10 patients with C. difficile icillin, performed by Maastricht University stimulate the production of cytokine se- infection (CDI), of whom 5 experienced Medical Centre, The Netherlands1. The trial cretory IgA (sIgA), an important immuno- recurrent CDI, twice-daily supplementa- showed that Ecologic® AAD significantly globulin for the general defence against tion with Ecologic® AAD, besides antibiot- reduced the risk of diarrhoea-like def- pathogens. ics, resulted in complete recovery4. ecation (figure 1). It was also shown that This result has been confirmed in a user Ecologic® AAD is recommended by the the intestinal microbiota of participants trial in Austria with 199 participants. Hos- World Gastroenterology Organisation in in the probiotic group restored faster to pitalised patients on antibiotics received their Global Guidelines (feb 2017).5 The the pre-antibiotic state than the intestinal Ecologic® AAD which resulted in a signifi- level of evidence on reducing antibiot- microbiota of participants in the placebo cantly lower incidence of AAD. Less than ic-associated diarrhoea is evaluated as group2. The results therefore suggest that 1% in the entire study group developed level 2 evidence: Randomized trial or restoration of the intestinal microbiota is AAD3, which normally ranges between observational study with dramatic effect.

Figure 1: Diarrhea-like defecation occurred significantly less in Figure 2: After day 35 there is a significantly faster recovery to the pre-antibiotic state of the the Ecologic® AAD group compared to the placebo group. microbiota in the Ecologic® AAD group compared to the placebo group.

90 * 100

80 90

70 80

60 70

50 60 50 40

79% Similarity (%) 40

Diarrhoea (%) 30 30 20 48% 20 10 10 Ecologic® AAD 0 0 Placebo 0 3 7 14 35 63 Days Ecologic® AAD Placebo

References Ecologic® AAD publications

1. Koning C.J.M et al. The Effect of a Multispecies Probiotic on the Intestinal Microbiota 3. Lang F.C. Use of a multi-species probiotics for the prevention of antibiotic associated and Bowel Movements in Healthy Volunteers Taking the Antibiotic Amoxycillin. Am J diarrhea. Nutrafoods 2010;9(2); 27-31. Gastroenterol 2007;102:1–12. 4. Hell M, et al. Probiotics in Clostridium difficile infection: reviewing the need for a multist- 2. Koning C.J.M. Multispecies probiotics and antibioticsassociated side effects. PhD Thesis. rain probiotic.Benef Microbes. 2013;4(1):39-51. 2010. 5. WAO Guidelines, Probiotics and Prebiotics Feb 2017. Reducing constipation

Strain selection Bacterial strains

The probiotic strains in Ecologic® RELIEF have been selected for their Ecologic® RELIEF is a multispecies formulation consisting of 7 probiotic capacity to inhibit pathogen growth and improve motility and per- strains, 1 x 109 CFU/gram. The recommended dosage is 4 grams a day. istalsis of the intestine. The strains have been screened in vitro for • B. bifidum W23 • L. casei W79 their capacity to: • B. lactis W51 • L. plantarum W62 • inhibit Clostridium difficile and Staphylococcus aureus • B. lactis W52 • L. rhamnosus W71 • produce lactic acid. • B. longum W108

Clinical evidence

Ecologic® RELIEF has been tested in a In addition, Ecologic® RELIEF has been cant increase in stool frequency; from 3.1 clinical study, in collaboration with the tested in an open-label study again at baseline to 6.7 in week four (p<0.01). Children’s Hospital of the Academic in collaboration with the AMC. During In addition, compared to baseline, a sig- Medical Centre (AMC) of the Universi- this study, 20 pregnant women suffer- nificant decrease was found in ty of Amsterdam. Twenty children suf- ing from constipation as defined by the - straining (35%), fering from constipation used Ecolog- Rome III criteria consumed 4 grams (1x109 - sensation of incomplete evacuation ic® RELIEF for four weeks (once daily). CFU/gram) Ecologic® RELIEF daily. Partic- (50%), Results showed that Ecologic® RELIEF ipants recorded daily amongst others, - sensation of anorectal obstruction significantly increased amount of bowel stool frequency and consistency, strain- (50%) movements per week, and decreased ing, sensation of incomplete defecation, - abdominal pain. amount of abdominal pain and faecal abdominal pain and adverse events in No adverse effects were reported2. incontinence (dirty underwear). No side a standardized bowel diary. Intake of effects were reported1 (see figure 1). Ecologic® RELIEF resulted in a signifi-

Figure 1: Ecologic® RELIEF is able to significantly increase bowel movements (A), decrease faecal incontinence (B) and decrease abdominal pain (C).

A. Bowel movements (toilet visits/week) B. Faecal incontinence (dirty underwear/week) C. Abdominal pain

P=0.13 P=0.001 P=0.10 P=0.007 Start 40% 20 16 15% 14 45%

12 No 10 Yes 8 Sometimes 10 25% After 6 4 weeks 55% 20% 4 Bowel movements/week Faecal incontinence/week Faecal 2

0 0 Baseline 2 4 Weeks Baseline 2 4 Weeks

References Ecologic® RELIEF publications

1. Bekkali N. et al. The role of a probiotics mixture in the treatment of childhood constipation: a pilot study. Nutr j 2007;6:17. 2. De Milliano I. et al. Is a multispecies probiotic mixture effective in constipation during pregnancy? A pilot study Nutr J 2012, 11:80. Reducing exercise-induced stress

Strain selection Bacterial strains

The bacterial strains in Ecologic® PERFORMANCE have been selected Ecologic® PERFORMANCE is a multispecies formulation consisting of for their anti-inflammatory and oxidative properties and their abil- 6 probiotic strains, 2,5 x 109 CFU/gram. The recommended dosage ity to improve the intestinal barrier function. The strains have been is 2 grams, twice daily. screened in vitro for their capacity to: • B. bifidum W23 • L. brevis W63 • strengthen the epithelial barrier • B. lactis W51 • Lc. lactis W58 • reduce oxidative stress • E. faecium W54 • induce the production of anti-inflammatory cytokines. • L. acidophilus W22

Clinical evidence

Ecologic® PERFORMANCE has been test- marker TNF-α (figure 1) and in the protein one or more URTI symptoms during the ed in a randomized, double-blinded, oxidation marker Carbonyl protein1. study period was 2.2-fold higher in the pla- placebo controlled trial at the Medical cebo group than in the probiotics group University of Graz, Austria. Twenty-three In a randomized, double-blinded, pla- (p=0.016)2 (figure 2). Also, the athletes in the endurance sportsmen received Ecologic® cebo controlled trial among 33 trained Ecologic® PERFORMANCE group could PERFORMANCE or placebo for 14 weeks. athletes at the Medical University of Inns- train 2.6 hours more per week compared Ecologic® PERFORMANCE significantly bruck, Ecologic® PERFORMANCE showed to the control group (p<0.001)2. These re- decreased stool zonulin levels (p=0,019), to significantly reduce the incidence sults show that Ecologic® PERFORMANCE an important marker for a ‘leaky gut’. of upper respiratory tract infections could reduce the adverse effects of stress Moreover, a trend was shown towards (URTIs). After 12 weeks of treatment, the and offer support during heavy exercise a decrease in the chronic inflammatory proportion of subjects who experienced training periods.

Figure 1: Effects of Ecologic® PERFORMANCE on TNF-α concentration. Figure 2. Incidence of URTIs before and after 12 weeks of supplementation with * After 14 weeks TNF-α was reduced by a trend in the Ecologic® PERFORMANCE Ecologic® PERFORMANCE. *Significant difference (p=0.016). group compared to the placebo group (p=0.054).

* Ecologic® PERFORMANCE pre stress test ® Ecologic pg/ml PERFORMANCE post stress test Placebo; ® pre stress test Ecologic

Number of individuals with URTI Placebo; PERFORMANCE post stress test Placebo Week 0 Week 14 Baseline Week 12

References Ecologic® PERFORMANCE publications

1. Lamprecht M, et al. Probiotic supplementation affects markers of intestinal 2. Strasser B, et al. Probiotic Supplements Beneficially Affect Tryptophan– barrier, oxidation, and inflammation, before and after intense exercise; Kynurenine Metabolism and Reduce the Incidence of Upper Respiratory a randomized, double-blinded, placebo-controlled trial. J Int Soc Sports Tract Infections in Trained Athletes: A Randomized, Double-Blinded, Nutr. 2012;9:45. Placebo-Controlled Trial. Nutrients. 2016;8(11) Preventing traveller’s diarrhoea

Strain selection Bacterial strains

The probiotic strains in Winclove Travel have been selected for their Winclove Travel is a multispecies formulation consisting of 8 pro capacity to protect against pathogens known to cause traveller’s biotic strains, 1 x 109 CFU/gram. The recommended dosage is 5 diarrhoea, such as; E.coli, Shigella and Salmonella, through: grams a day. It is advised to start supplementation a couple of days • producing antimicrobial agents before travel until the end of the journey. • inhibiting pathogen adhesion • B. bifidum W23 • L. casei W56 • L. salivarius W24 • competing with pathogens for nutrients. • B. lactis W51 • L. plantarum W21 • Lc. lactis W58 • L. acidophilus W37 • L. rhamnosus W71

Clinical evidence

Winclove Travel has been tested in a The effect of individual, as well as com- be seen in figure 2, single strains reduced pilot study with 200 travellers travelling binations of Winclove Travel strains on the invasion ability of E.coli, however, to high-risk areas. The travellers took 5 five intestinal pathogens (S. enteritidis, L. strain combinations induced an even grams of Winclove Travel once daily, monocytogenes, E. coli O157:H7 (ETEC), more remarkable effect. These results starting a couple of days before their trip. C. sakazakii, C. jejuni) has been tested in confirm the importance of multispecies All travellers filled out a diary during their vitro.1 All the strains showed antagonistic formulations to potentiate beneficial ef- trip, scoring their defecation pattern ac- effects against the pathogens through fects and indicate that supplementation cording to the Bristol stool scale. Results different inhibitory mechanisms, includ- with Winclove Travel could prevent TD. show that Winclove Travel is able to re- ing; antimicrobial activity, interference of duce the risk of TD. Only 19% developed adhesion of the pathogens to the intes- TD, which is normally 40%. Of the partici- tinal wall, and growth competition/ex- pants 42% stated to have benefited from clusion. Combinations of strains showed the formulation (figure 1). greater effect than single strains. As can

Figure 1: Clinical evidence of 200 travellers Figure 2: Invasion inhibition of ETEC by single and combinations of Winclove Travel strains measured in an exclusion taking Winclove Travel. and competition test. A decrease in CFUs (expressed on a log scale) indicates inhibition. * Significant effect (P <0.05). Exclusion test Competition test

19% 39%

42%

log CFU/ml

Stated to have benefited from Winclove Travel No effect Developed TD ETEC ETEC + W23 ETEC + W24 ETEC + W7 ETEC + ETEC + ETEC + ETEC + W23 + W23 + W24 W23 + W71 W24 + W71 W24 + W71

References Winclove Travel publication

1. Campana R. et al. Strain-specific probiotic properties of lactic acid bacteria and their interference with humanintestinal pathogens invasion. Gut Pathog. 2017;9(12). Improving intestinal health in newborns

Strain selection Bacterial strains

The probiotic strains in Winclove Baby are active on all three levels in Winclove Baby is a broad-spectrum, multispecies formulation con- the gut (external, barrier, systemic). The strains have been screened sisting of 4 probiotic strains, 1 x 109 CFU/gram. The recommended in vitro for their capacity to: daily dosage is variable. • inhibit various pathogens such as; C. difficile and E. coli • B. animalis W12 • B. longum W108 • improve the barrier function • B. breve W25 • influence the immune system. • B. lactis W51

In vitro evidence

The most dominant genera in the healthy Maintenance of the intestinal barrier func- Disruptions in the early microbiota can infant’s gut is Bifidobacterium. Therefore, tion is critical for essential physiological cause an imbalance in immune response Winclove Baby consists of four Bifido- processes. Three strains of Winclove Baby regulating cells (Th1/Th2), which is one of bacterium species, of which research have been added for their ability to pro- the main mechanisms behind the devel- has shown to be present in the gut of tect the integrity of the intestinal lining. opment of allergies and asthma. All the infants3. These Bifidobacterium spe- Their efficacy has been tested in a Trans strains in Winclove Baby are selected and cies can restore the balance of the Epithelial Electrical Resistance (TEER) mod- tested to maintain a balanced immune new-born’s microbiota. Three strains of el. All strains were capable to strength- system by inducing the production of IL- Winclove Baby have been added en the intestinal barrier function when a 10; a cytokine with anti-inflammatory ef- for their ability to inhibit growth of E. stressor was added. (figure 2). fects. (figure 3). coli, a pathogen associated with infant colic. These strains have shown good capability to inhibit growth of E. coli (figure 1).

Figure 1: Inhibition of E.coli by three strains in Winclove Baby. Figure 2: TEER data of three strains in Winclove Baby. Figure 3: Production of the anti-inflammatory cytokine IL-10 The larger the zone, the better the inhibition. The higher %TEER, the better the barrier function. after 24hr by the strains in Winclove Baby.

% TEER

Inhibition zone (mm) Inhibition zone

IL-10 production after production (pg/ml) 24hr IL-10

Control B. animalis B. lactis B.longum Control Stressor Stressor+ Stressor+ Stressor+ Control B. animalis B. breve B. lactis B. longum W12 W51 W108 B. animalis B. breve B. lactis W12 W25 W51 W108 W12 W25 W51 Improving intestinal health in children

Strain selection Bacterial strains

The probiotic strains in Winclove Junior are active on all three levels in Winclove Junior is a broad-spectrum, multispecies formulation con- the gut (external, barrier, systemic). The strains have been screened in sisting of 6 probiotic strains, 5 x 108 CFU/gram. The recommended vitro for their capacity to: daily dosage is variable. • inhibit various pathogens such as; C. difficile, E. coli, Salmonella, • B. lactis W51 Shigella and P. agglomerans • B. lactis W52 • improve the intestinal barrier function • L. acidophilus W55 • stimulate a less allergic response of the immune system by; induc- • L. casei W56 ing T-and B-cell proliferation, increasing IFN-y and IL-10 production • L. salivarius W57 and decreasing IL-5 and IgE production. • Lc. lactis W58

Clinical evidence

In a post-marketing study among thera- Figure 1: Results of a post-marketing survey with Winclove Junior. pists in the Netherlands it was investigat- 100 ed for which indications Winclove Junior 80 is prescribed and how its use is experienced (positive/negative). The results of 60 this survey are shown in figure 1. As can 40 be seen in the graph, Winclove Junior is Positive results (%) Positive results 20 prescribed for many health problems and shows the most satisfying effect (100%) 0 Antibiotics IBS Food Constipation Food Respiratory IBD Allergy Diarrhoea for use during antibiotic use. Also for the Intolerance Infections Tract other indications, high satisfaction scores Infections (60%-80%) were obtained (data not published).

In addition, Winclove Junior has been Figure 2: Results of a post-marketing survey with Winclove Junior. * Significant effect. studied in another post-marketing survey investigating the effect of Winclove Junior on childrens’ health. 80 children in the age of 0.5 – 6 years old used the probiotic for 6 weeks. A general health questionnaire was filled out by their healthcare profes- Positive results (%) Positive results sional together with the parent(s). Results showed that Winclove Junior significantly increased health and improved appetite, Health as Mood Sleep Playfulness Crying and Appetite Stool Stool Flatulance indicated rhythm wining intensity frequence composition see figure 2. The researchers reported no side-effects (data not published). Start 3 weeks 6 weeks Improving intestinal health in adults

Strain selection Bacterial strains

The probiotic strains in Winclove Adult are active on all three levels in Winclove Adult is a broad-spectrum, multispecies formulation con- the gut (external, barrier, systemic). The strains have been screened sisting of 8 probiotic strains, 1 x 109 CFU/gram. The recommended in vitro for their capacity to: daily dosage is variable. • inhibit various pathogens such as; C. difficile, E. coli, E. faecalis and • B lactis W51 • L. paracasei W20 B. subtilus. • B. lactis W52 • L. plantarum W21 • improve the barrier function • E. faecium W54 • L. salivarius W24 • influence the immune system. • L. acidophilus W22 • Lc. lactis W19

Clinical and in vitro evidence

It has been shown in vitro that the The effect of Winclove Adult on gut health tom significantly increased with 27.5% strains in Winclove Adult are able to and quality of life (QoL) was tested in (p<0.02) for gastrointestinal pain, 25% inhibit different pathogenic species an observational study. 40 adults with (p<0.03) for epigastric fullness/bloating such as; E. coli, E. faecalis and B. subti- GI-complaints (but otherwise healthy) and 20% (p<0.02) for flatulence (see figure lis. The strains have also shown to in- were supplemented for a 6-week pe- 2). There was an up-going trend in these hibit Clostridium difficile, the pathogen riod with Winclove Adult. The results scores, suggesting that further improve- which often causes diarrhea after the show a significant reduction in the total ment could be expected on prolonged use of antibiotics. In addition, the com- GI-symptom score from 10 at baseline to use of Winclove Adult. At the start of the plete formulation was tested on its abil- 6 at the end of the intervention (p<0.05)1. study participants already reported a high ity to inhibit E. coli and Salmonella and The percentage of participants which quality of life; a slight improvement after 6 Shigella species (figure 1). were fully resolved of a particular symp- weeks of supplementation was seen.

Figure 1: Inhibition of E.coli, Salmonella and Shigella by Figure 2: Percentage of participants that were asymptomatic for epigastric pain, epigastric fullness and flatu- Winclove Adult, tested in an in vitro well diffusion test. lence after 6 weeks supplementation with Winclove Adult. * Significant increase (p<0.05). The larger the inhibition zone, the better the inhibition.

Epigastric pain Epigastric fullness Flatulence

that are asymptomatic that are Inhibition zone (mm) Inhibition zone

Percentage (%) of participants

Baseline week 0 week 1 week 2 week 3 week 4 week 5 week 6 Control E.coli Shigella sp. Salmonella sp.

References Winclove Adult publication

1. Grossenbacher F, Gashi A, Besseling-van der Vaart I. Use of the multispecies probiotic Winclove 500/Bactosan pro FOS leads to less gastrointestinal complaints in adults - an observational in vivo pilot study. Advances in Microbiology 2016; 6 (14): 975-985. Improving intestinal health in elderly

Strain selection Bacterial strains

The probiotic strains in Winclove Senior are active on all three levels in Winclove Senior is a broad-spectrum, multispecies formulation con- the gut (external, barrier, systemic). The strains have been screened in sisting of 10 probiotic strains, 1 x 109 CFU/gram. The recommended vitro for their capacity to: daily dosage is variable. • inhibit various pathogens such as; E. coli, Salmonella, Shigella and • B. animalis W53 • L. plantarum W1 P. agglomerans • B. bifidum W23 • L. rhamnosus W71 • improve the intestinal barrier function • B. lactis W51 • L. salivarius W24 • influence the immune system. • B. lactis W52 • Lc. lactis W19 • L. acidophilus W22 and vitamin D3 • L. paracasei W20

In vitro evidence

Winclove Senior contains 10 bacterial strains. In addition, vitamin D3 has been added to the formulation. Vitamin D contributes, among others, to the maintenance of normal bones, teeth and muscle function. Vitamin D status is often impaired in elderly. Winclove Senior has been tested in vitro on:

• Inhibition of Clostridium difficile. • Strengthening the barrier function. • Boosting the immune system. Elderly are more susceptible to C. difficile Ageing comes with an impaired intesti- The immune system gradually decreases infections. Three bacterial strains have nal barrier function. Three bacterial strains in older people. Three bacterial strains been added to the formulation to inhibit have been added to strengthen the intes- have been added to the formulation to growth of C. difficile. In vitro testing has tinal barrier function. The results of in vitro boost the immune system. These strains shown excellent capability of these strains testing with two strains in a Trans Epithe- have shown good induction of Th1 cells, to inhibit C. difficile, see figure 1. lial Electrical Resistance (TEER) model are which are involved in promoting immune shown in figure 2. Both strains are very responses and defence. (Results not well capable of strengthening the intes- shown). tinal barrier function. The third strain also performed well in a different model.

Figure 1: The inhibition of C. difficile and its toxins by three bacterial strains in Figure 2: The capacity of two bacterial strains in Winclove Senior to strengthen the Winclove Senior. The lower the concentration, the better the inhibition intestinal barrier function. The higher %TEER, the better the barrier function.

C. difficile after 48hr Toxins after 48hr (log cfu/ml)

C. difficile % TEER Concentration Control L. paracasei L. salivarius L. rhamnosus Control stressor stressor + stressor + W20 W24 W71 B. bifdum W23 B. animalis W53 Interested? Don’t hesistate to contact us, we look forward to hearing from you!

PROBIOTICS

Winclove Probiotics Hulstweg 11 1032 LB Amsterdam, The Netherlands +31 (0)20-435 02 35 [email protected] www.wincloveprobiotics.com

All probiotic strains have the Qualified Presumption of Safety Status (QPS). Winclove is a NSF International Certified GMP facility for manufacturing dietary supplements. Winclove’s food safety management system is ISO 22000:2005 certified for the development and production of prebiotics and probiotics. Both our formulations and manufacturing processes are controlled by The information in this brochure is intended for business certified, external laboratories. professionals only and should not be given to consumers.