COMBINATION OF LACTOBACILLI FOR THE RELIEF OF IRRITABLE BOWEL SYNDROME AND FOR THE RELIEF OF OTHER GASTROINTESTINAL DISORDERS.

Abstract

This description outlines compositions and methods related to the use of a combination of live lactobacillus bacteria, particularly live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus, for the relief of undesirable gastrointestinal health problems such as irritable bowel syndrome (IBS), abdominal pain, abdominal discomfort, bloating, loose stools, and constipation. This combination of live lactobacillus bacteria can also be used to improve the quality of life of a subject suffering from IBS, for the relief of IBS, and / or for the prevention and / or treatment of IBS.

Description

Combination of lactobacilli for the relief of irritable bowel syndrome and for the relief of other gastrointestinal disorders FIELD OF INVENTION The invention relates to the field of gastrointestinal disorders, and more particularly to the use of a combination of live lactobacilli bacteria for the relief of undesirable gastrointestinal health problems such as irritable bowel syndrome (IBS), abdominal pain, abdominal discomfort, abdominal distension, loose stools, and constipation. BACKGROUND OF THE INVENTION Irritable bowel syndrome (IBS) is a chronic, relapsing gastrointestinal disorder that affects 5–20% of the American population. Several risk factors for IBS have been identified, including female sex, psychological problems, stress, food intolerances, and small intestinal bacterial overgrowth (SIBO) (Aagaard et al., 2013). Cardinal symptoms of IBS include abdominal pain, bloating, and changes in bowel habits (Aagaard et al., 2013). The pathophysiology is well-defined, and there are no associated structural intestinal abnormalities. The quality of life of individuals with IBS is severely impaired, with significant impacts on the healthcare system and visits to primary care physicians and gastroenterologists (Coffin et al., 2004).In fact, IBS is the most frequent diagnosis in gastroenterology clinics and one of the most frequent diagnoses in primary care (Peery et al., 2012). Based on specific symptoms, patients with IBS can be subclassified into three main groups: constipation-predominant (IBS-C), diarrhea-predominant (IBS-D), and mixed bowel patterns (IBS-M), each with a roughly equal distribution. These symptoms of IBS are uncomfortable for patients, result in a lower quality of life, and interfere with social interactions (Coffin et al., 2004). The ultimate goal of SIL treatment is to provide relief from the multiple symptoms of this condition using a single, well-tolerated agent. Drug therapies can alleviate some of the symptoms associated with this condition, but none are curative. Therefore, the outlook for long-term treatment efficacy is limited given the available options. Current treatments. There is a clear need for Sil relief procedures that are safe, effective, and cost-effective (Foxx-Orenstein, 2006). Probiotics are live microorganisms that provide health benefits to the host when administered in adequate doses. In recent years, probiotics have been commonly used to alleviate symptoms in a variety of gastrointestinal disorders. Since dysbiosis can be part of the multifactorial etiology of SIL, a variety of probiotics have been tested in clinical trials to determine their efficacy, and the results have been included in several meta-analyses and review articles (Ford et al., 2014b; Hoveyda et al., 2009; McFarland and Dublin, 2008; Ortiz-Lucas et al., 2013; Whelan and Myers, 2010; Yoon et al., 2015).No firm conclusions can be drawn about the effectiveness of strain-specific probiotics for SIL symptom relief. Strong placebo effects, psychological factors, and gender effects make it difficult to interpret the study findings (Ford & Moayyedi, 2010; Lyra et al., 2016; Moayyedi et al., 2010). Probiotic products containing lactobacilli and bifidobacteria have already been tested for improving ILS (Niv et al., 2005; O'Mahony et al., 2005). Positive results have been observed with some Lactobacillus strains, for example, by Ducrotté et al., who reported the resolution of all dominant ILS symptoms, including abdominal pain, in 214 patients treated for 4 weeks with L. plantarum 299V (Ducrotté et al., 2012). Halpern et al. noted a significant reduction in an ILS symptom index with a capsule containing 5 x 10⁹ heat-killed L. acidophilus (Halpern et al., 1996). Other strains of Lactobacillus such as L. salivaria UCC4331 showed no therapeutic gain over placebo in 75 patients (O'Mahony et al., 2005), nor did L. reuteri ATCC55730 (Niv et al., 2005), suggesting that some strains of Lactobacillus may be more effective than others in this indication. Although products comprising strains of Lactobacillus acidophilus, Lactobacillus casei and / or Lactobacillus rhamnosus have been tested during clinical research in the past (Beausoleil et al., 2007; Gao et al., 2010; Maziade et al., 2015; Sampalis et al., 2010), these clinical trials never showed or suggested the effectiveness of lactobacilli in relieving irritable bowel syndrome (IBS), abdominal pain, bloating and / or constipation. Consequently, there is a need for a combination of bacterial strains that are effective in relieving gastrointestinal disorders such as irritable bowel syndrome (IBS). LCRQQn / nznz / q / YiAi abdominal pain, abdominal distension and / or constipation. The present invention addresses these and other needs, as will become evident from a review of the disclosure and description of the features of the invention below. BRIEF SUMMARY OF THE INVENTION The invention relates to the use of a combination of live lactobacilli bacteria for the relief of undesirable gastrointestinal health problems such as irritable bowel syndrome (IBS), abdominal pain, abdominal discomfort, abdominal distension, loose stools, and constipation. According to one particular aspect, the invention relates to a method for relieving a gastrointestinal disorder in a subject in need, comprising administering to said subject a combination of live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus, wherein said gastrointestinal disorder is selected from the group consisting of irritable bowel syndrome (IBS), abdominal pain, abdominal distension, and constipation. According to another particular aspect, the invention relates to a method for improving the quality of life of a subject suffering from irritable bowel syndrome (IBS), comprising administering to said subject a combination of live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus, wherein said administration provides said subject with at least one benefit selected from the group consisting of: satisfaction with bowel habits, minimal interference of IBS with normal activity, improved body image, reduced food avoidance, increased social reaction, reduced sexual dysfunction, and improved relationships. According to another particular aspect, the invention relates to a method for the relief of irritable bowel syndrome (IBS) in a human subject in need, comprising administering to said subject a combination of live Lactobacillus acidophilus CL1285®, live Lactobacillus casei LBC80R® and live Lactobacillus rhamnosus CLR2®. According to another particular aspect, the invention relates to an alternative method to drug therapy for the prevention and / or treatment of irritable bowel syndrome (IBS), comprising: (i) identifying a human subject who requires drug therapy for the prevention and / or treatment of IBS; and (ii) administering to said human subject a nutritionally acceptable composition comprising a combination of live microorganisms comprising live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus LCRQQn / nznz / q / YiAi live, in addition to or as a substitute for such drug therapy. According to another particular aspect, the invention relates to a composition for the relief of a gastrointestinal disorder in a subject in need, the composition comprising a combination of live Lactobacillus acidophilus CL1285®, live Lactobacillus casei LBC80R®, and live Lactobacillus rhamnosus CLR2®, wherein said gastrointestinal disorder is selected from the group consisting of irritable bowel syndrome (IBS), abdominal pain, days of pain, distension, stool consistency, and stool frequency. According to another particular aspect, the invention relates to a composition for the relief of irritable bowel syndrome (IBS) in a subject in need, the composition comprising a combination of live Lactobacillus acidophilus CL1285®, live Lactobacillus casei LBC80R® and live Lactobacillus rhamnosus CLR2®. According to another particular aspect, the invention relates to the use of a combination of live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus, for the relief of a gastrointestinal disorder in a subject in need, wherein said gastrointestinal disorder is selected from the group consisting of irritable bowel syndrome (IBS), abdominal pain, abdominal distension, and constipation. According to another particular aspect, the invention relates to the use of a combination of live Lactobacillus acidophilus CL1285®, live Lactobacillus casei LBC80R® and live Lactobacillus rhamnosus CLR2®, for the prevention, treatment and / or relief of irritable bowel syndrome (IBS) in a subject who needs it. DETAILED DESCRIPTION OF THE PREFERRED MODALITIES The invention relates to the prevention, treatment and / or relief of gastrointestinal disorders in subjects. As used herein, the term “gastrointestinal disorder” refers to gastrointestinal disorders characterized by symptoms such as abdominal pain, bloating, and constipation (e.g., stool consistency and frequency). The term encompasses, but is not limited to, irritable bowel syndrome (IBS), including constipation-predominant IBS (IBS-C), diarrhea-predominant IBS (IBS-D), and mixed-bowel-pattern IBS (IBS-M). As used herein, the term “relief from a gastrointestinal disorder” or “relief from irritable bowel syndrome” or “IBS relief” encompasses health benefits that include, 4 LCRQon / nznz / q / YiAi but are not limited to, stabilizing, curing, healing, mitigating, relieving, altering, remedying, lessening the worsening, recovering, improving, or affecting the disease or condition, the symptom of the disease or condition, or the risk of (or susceptibility to) the disease or condition. The term encompasses at least the relief of undesirable health problems which include, but are not limited to, abdominal pain, prolonged duration of abdominal pain (consecutive or not), abdominal distension, problems with stool consistency and / or frequency (e.g., constipation), reduced quality of life (QOL) associated with one or more of the foregoing, and inadequate relief of such health problems when treated with drugs or other means. As used herein, the term “subject” includes living organisms in which gastrointestinal disorders may occur. The term “subject” includes animals (e.g., mammals (e.g., cats, dogs, horses, pigs, cows, goats, sheep, rodents (e.g., mice or rats), rabbits, squirrels, bears, primates (e.g., chimpanzees, monkeys, gorillas, and humans)), as well as birds (e.g., chickens, ducks, Pekin ducks, geese), and transgenic species thereof. Preferably, the subject is a human or non-human primate (e.g., chimpanzee, monkey, macaque, gorilla). More preferably, the subject is a human. Even more preferably, the subject is a human in need of treatment who has, or is likely to have, one of the most undesirable health problems and / or symptoms, such as abdominal pain, bloating, and constipation. According to one particular aspect, the invention provides the use of a combination of live Lactobacilli for the prevention, treatment and / or relief of gastrointestinal disorders, particularly SIL. According to one particular embodiment, the combination of live Lactobacilli comprises live Lactobacillus acidophilus, live Lactobacillus casei and live Lactobacillus rhamnosus. In the modalities, the combination comprises approximately 1-10% of L. acidophilus, 70-90% of L. casei and approximately 5-20% of L. rhamnosus of the colony forming units (CFU) of the combination. In a particular modality, Lactobacillus acidophilus is Lactobacillus acidophilus CL1285® deposited in the National Collection of Cultured Microorganisms (CNCM) in Paris, deposit No. CNCM 1-4099, Lactobacillus casei is Lactobacillus casei LBC80R® deposited in the CNCM with deposit No. CNCM 1-3989, and Lactobacillus rhamnosus is Lactobacillus rhamnosus CLR2® deposited in the CNCM as deposit No. CNCM 1-3990. In the modalities, the invention comprises simultaneously administering at least 10 LCRQQn / nznz / q / YiAi billions, or at least 20 billion, or at least 30 billion, or at least 40 billion, or at least 50 billion, or at least 75 billion, or at least 100 billion, or at least 150 billion, or at least 200 billion of said combination of Lactobacilli. In the modalities, the invention comprises administering the combination of live Lactobacilli once a day, twice a day, three times a day or more. In these formulations, the combination of live Lactobacilli is administered as a nutritionally acceptable composition. As used herein, the term nutritionally acceptable composition refers to a substance, for example, a food substance, that, when ingested, will provide nutritional support and nutrients such as carbohydrates, fats, proteins, vitamins, and / or minerals, etc. Once ingested, in addition to providing health benefits (for example, relief from undesirable gastrointestinal problems), the nutritionally acceptable composition will also provide energy, like any other food substance.A nutritionally acceptable composition according to the invention is substantially different from compositions used in drug therapy; the nutritionally acceptable composition is composed of food ingredients (preferably natural ingredients) that are recognized as safe, non-toxic to humans, and substantially free of the side effects associated with typical prescription drugs (e.g., headache, nausea, allergy, etc.). However, the nutritionally acceptable composition may also include additional safe and non-toxic components such as preservatives, solubilizing agents, stabilizing agents, emulsifying agents, softening agents, coloring agents, odor agents, antioxidants, etc. The nutritionally acceptable composition may be presented as a solid, dry, or liquid form for oral administration. It may be presented in a variety of ingestible forms of food or dietary supplements, including, but not limited to, milk, yogurt, curd, fermented milks, fermented milk products, fermented soy products, fermented cereal products, powdered milk products, infant formulas, protein concentrates such as those used in hospitals, etc. In the modalities, the nutritionally acceptable composition comprises the combination of Lactobacilli and also fermented proteins which includes, but is not limited to, fermented soy proteins, fermented milk proteins, fermented rice proteins, pea proteins 6 LCRQon / nznz / q / YiAi fermented, fermented hemp proteins, fermented almond proteins and fermented insect proteins (e.g. larval proteins). The combination of Lactobacilli and the nutritionally acceptable composition can be incorporated into any suitable oral dosage form, such as a gel, capsule, tablet, suspension, or any other suitable form known to those skilled in the art. Preferably, the quantity of Lactobacilli contained in a single capsule, tablet, suspension, or similar product is in the range of approximately 10 trillion to 200 trillion. The invention also encompasses cases and containers comprising multiple doses of the nutritionally acceptable composition, including, for example, blister packs, resealable bottles, and the like, comprising a certain quantity of the composition (e.g., 25 mL, 50 mL, 100 mL, or more) or a number of capsules or tablets (e.g., 10, 15, 25, 50, or more). Such a case or container may advantageously include instructions in the form of a leaflet or other printed material, indicating the quantities of the composition to be administered, instructions for administration, instructions for mixing the components (e.g., if in powder form), etc. The manufacture of a nutritionally acceptable composition according to the invention is within the capabilities of those skilled in the art. For example, Lactobacilli can be incorporated into a suitable, nutritionally acceptable carrier. Alternatively, a nutritionally acceptable composition comprising the combination of Lactobacilli can be obtained by fermenting live Lactobacilli bacteria in a suitable medium to obtain a ferment comprising the Lactobacilli and fermented proteins (e.g., fermented soy protein, fermented milk protein, fermented rice protein, fermented pea protein, fermented hemp protein, etc.). Example The following example serves to illustrate the scope of use of the present invention and not to limit its scope. Modifications and variations may be made without forgetting the intent and extent of the invention. Although other methods or products equivalent to those described herein may be used to test or to carry out the present invention, the preferred materials and methods are described. Example 1: Lactobacillus acidophilus CL1285®, L. casei LBC80R® and L. rhamnosus CLR2® improve quality of life and symptoms of SILC, IBS-D: Double-blind, randomized, placebo-controlled study The objectives of this clinical trial were to evaluate the effectiveness of a patented probiotic product, Lactobacillus acidophilus CL1285® + Lactobacillus casei LBC80R® + Lactobacillus rhamnosus CLR2®, for the relief of specific IBS-related symptoms, improvement in quality of life, effect on stool consistency and frequency, and achievement of adequate relief (AR) in otherwise healthy adults with irritable bowel syndrome of the constipation (IBS-C), diarrhea (IBS-D), and mixed (IBS-M) subtypes. Materials and methods Experimental design, study implementation, and data collection The protocol for this prospective, double-blind, randomized, placebo-controlled study was approved by an independent review board, IntegReview. All participating subjects provided written informed consent. Subjects aged 18 years or older were recruited from three clinical trial sites located in California, USA. The subjects ingested two capsules of either the active product or the placebo with breakfast each day. Each active capsule contained 50 billion units of live organisms (L. acidophilus CL1285®, L. casei LBC80®, and L. rhamnosus CLR2®) plus inert ingredients. The placebo capsules contained only inert ingredients. Subjects were required to meet the Rome III criteria for SIL (Shih and Kwan, 2007). The Rome III criteria include the presence of recurrent abdominal pain or discomfort at least 3 days / month in the past 3 months, associated with 2 or more of the following: improvement with defecation, onset associated with a change in stool frequency, and onset associated with a change in stool form (appearance). The onset of symptoms must be at least 6 months prior to diagnosis. Subjects were required to complete a 7-day placebo pre-enrollment period to demonstrate adherence to investigational product (IP) intake and completion of diaries documenting IP consumption, stool frequency, stool consistency as defined by the Bristol Stool Chart (BSC), pain severity, and concomitant medications. Successful completion of the pre-enrollment period also required the presence of abdominal pain on at least 2 days, associated with at least 2 of the following: improvement with defecation, onset associated with a change in stool frequency, and onset associated with a change in stool shape or appearance. Potential subjects with diagnosed gastrointestinal disease other than subacute gastrointestinal syndrome (SGS) or abdominal surgery were excluded. LCRQQn / nznz / q / YiAi Previous or systemic disease with the potential to confound study results or compromise safety, life expectancy less than 6 months, pregnancy or lactation, lactose intolerance, immunodeficiency, eating disorder, recent antibiotic use, allergy to the study product, or daily consumption of probiotics, fermented milk, or yogurt. After successful completion of the pre-enrollment period, 113 subjects were randomized in a 2:1 ratio to either an active study product or placebo. The subjects returned to the study site at 6-week intervals for a total of 12 weeks of study. At each visit, the subjects completed two questionnaires, the Sll-SSS (Symptom Severity Scale) and the SII-QOL (Quality of Life Measure, which includes an overall score and an assessment of quality of life in eight validated domains: dysphoria, interference with activity, body image, health concern, food avoidance, social reaction, sexual, and relationship; see “Information Sheet on the Irritable Bowel Syndrome-Quality of Life Measure (SII-QOL)” published by the University of Washington, which is available online). At each visit, subjects were asked whether they had experienced adequate relief of SIL symptoms. Subjects continued to record stool consistency and frequency, symptom severity, PI consumption, and concomitant medications in diaries, which were collected at each visit and checked for legibility and completeness. The returned PI was counted to assess adherence, and a new PI was issued at visit 3. Subjects were asked about any adverse events (AEs) recorded in the diary to determine the dates of onset and recovery and severity. Subsequently, the investigator categorized the reported AEs according to their relationship to the PI (related, possibly related, unlikely to be related, unrelated). Study evaluation criteria The study endpoints included changes in abdominal pain score, bloating score, days with pain, improvements in IBS-SSS and IBSQOL scores (which includes quality of life domains), and AA. Changes in stool frequency and stool consistency during the study period were examined within IBS subtypes and within IBS subtype and gender subgroups. The safety endpoints were the incidence, severity, and relationship of PI to reported adverse events. Populations under study A modified intention-to-treat (mITT) population was defined as subjects who were 9 LCRQQn / nznz / q / YiAi were randomized and received at least one dose of the PI; this population was used for the efficacy analysis and the safety analysis. Data management Data were collected from printed source documents at the study sites and entered into a network-based relational database. Clinical research associates (CRAs) verified 100% of the clinical data fields against the source document on-site; queries were generated as needed for resolution by the site clinical team. Once all data were entered and all queries resolved, the database was locked for analysis. The study biostatistician then extracted the data files, and subject identification numbers were compared with their treatment assignments to clarify the study. Statistical analysis The number of subjects selected, the number randomized, the number of early withdrawals, and the number completed were tabulated by treatment group. The mITT population was analyzed as a whole for symptom and quality of life assessment criteria, along with Sil subtype and gender subpopulations. Changes in stool consistency and frequency were analyzed for SILE and SILD subtypes and by gender within each subtype. Descriptive statistics were calculated for baseline and demographic characteristics and tabulated by treatment group. Descriptive statistics include means, standard deviations, medians, ranges, and percentages, as dictated by the form of each variable. Inferential methods were not applied to baseline characteristics. Adherence was calculated as the percentage of the planned intake (PI) used, determined by the count of returned bottles and the subjects' diaries at weeks 6 and 12, and was compared between groups. Adherence was also defined as an intake of 70% or more of the planned PI and was analyzed using the chi-square test. Changes in scores between Visit 2 and Visit 4 were analyzed in both treatment groups for IBS-SSS, overall SILQOL and its domains, pain severity, days with pain in the past 10 days, bloating severity, bowel habit satisfaction, and IBS interference with daily life. Stool consistency scores were self-assessed by subjects using the BSC and recorded in their diaries on a daily basis, and daily stool frequency was determined from the number of stools. LCRQQn / nznz / q / YiAi entered into the diary. Changes in median stool consistency and stool frequency were compared during the 7-day pre-inclusion period versus the last 7 days of the study. Stool consistency scores were expressed as the median BSC scores per week, while stool frequency was expressed as the median number of stools per day. Data analysis revealed that efficacy endpoints had to be assessed within Sil subtypes and for each gender separately, and many of the subgroup sample sizes were small. A large placebo effect was observed for many endpoints. Therefore, we chose to control for the placebo effect by comparing changes in the Active versus Placebo groups. The mean improvement from Visit 2 to Visit 4 was calculated for each treatment group, and then the Placebo value was subtracted from the Active value, divided by the Placebo value, and multiplied by 100. For example, a mean change in pain severity of 15.0 in the Active group versus a mean change of 10.0 in the Placebo group was reported as a 50% improvement for the Active group over the Placebo group.This approach was used to compare changes in IBS-SSS, IBS-QOL, and related domains, pain severity, days with pain, bloating severity, bowel habit satisfaction, and interference of IBS with life in general. The same method was used to compare changes in stool consistency and frequency. The analysis of changes in stool consistency and frequency was conducted in subjects with the IBS-C and SILD subtypes, and in male and female subjects within those subtypes. Within each subtype, the percentage of “improvement” was defined as the percentage change in the direction appropriate for that subtype. Therefore, the results tables report “improvement” as a positive change for both subtypes, but the definition differs: for the IBS-C subtype, an increase in the mean BSC score (corresponding to softer stools) and an increase in stool frequency were positive scores indicating improvement for that endpoint. For subjects with SILD, a decrease in the mean BSC score (indicating firmer stools) and a decrease in stool frequency were reported as improvement using positive numbers. At the time the protocol was written, the AE (Assessment Approach) was a common primary endpoint in IBS trials and was adopted as the endpoint for this trial. The IBS-AE endpoint has been shown to be a clinically and statistically relevant benefit in therapeutic trials of IBS with alosetron (Camilleri et al., 1999), cilansetron, and tegaserod (Kellow et al., 2003; Tack et al., 2005). The AE consists of a single question: “During the past week, have you had adequate relief from your IBS symptoms?” Safety was assessed by calculating the rates of subjects with adverse events in the Active and Placebo groups and comparing them descriptively. Specific categories of adverse events were tabulated descriptively. Comparisons of subjects with specific adverse events were descriptive. Results A total of 113 subjects enrolled, of which 86 subjects (76.1%) completed the study. Termination rates were 73.0% in the placebo group and 77.6% in the active group.10 Reasons for early discontinuation included loss to follow-up (10.6%), withdrawal of consent (7.1%), and other / unknown reasons (6.1%). No subjects withdrew due to an adverse event. Demographic and baseline characteristics of the subject The distribution of demographic and baseline characteristics of the mITT population is presented in Table 1. The Placebo and Active groups were comparable in age, gender, and race. Table 1. Demographic and baseline characteristics of subjects at the screening visit, by treatment group, mITT population Placebo Active Age (years) Mean 39.9 40.6 SD 14.0 13.4 n 37 76 Sex (#, %) Male 16 (43.2%) 29 (38.2%) Female 21 (56.8%) 47 (61.8%) n 37 76 Race / ethnicity (#, %) Caucasian, non-Hispanic 14 (37.8%) 31 (40.8%) Asian 2 (5.4%) 3 (3.9%) Hispanic 7 (18.9%) 15 (19.7%) Native American 1 (2.7%) 0 Black or African American Other (35.1%) (32.9%) (2.6%) LCRQQn / nznz / q / Yi Distribution of Sil subtypes The 113 patients were classified by the researchers at each site as SILE, IBS-D, or IBS-M based on their symptoms and history at study entry. The distribution of subjects across the three subtypes varied by clinical site, as shown in Table 2. Table 2. Number and percentage of subjects in each Sil subtype by research site, mITT population SILE SILD SII-M Total Garden 12 (75.0%) 1 (6.3%) 3(18.6%) 16(100.0%) Grove San 12 (23.5%) 22 (43.1%) 18 (34.6%) 52 (100.0%) Francisco Westlake 16 (35.6%) 29 (64.4%) 0 45 (100.0%) Total 40 (35.7%) 52 (46.4%) 21 (18.6%) 113 (100.0%) Compliance Subjects in the placebo group consumed 87.0 ± 17.8% of the intended dose, while in the active group consumption was 77.3 ± 19.9%. According to the protocol, consumption of at least 70% of the intended dose was defined as adherence by 84.4% of subjects in the placebo group and 87.3% of subjects in the active group. Sil Symptom Severity Scale - SILSSS The SILSSS consists of questions about the severity of abdominal pain, the number of days with pain in the past 10 days, the severity of abdominal distension, satisfaction with bowel habits, and the degree to which the urinary tract interferes with the subject's life in general. All of these, except for the number of days with pain, were scored on a Likert scale with a range of 0–100. When the overall score was calculated, no mean improvement of 30% or more was demonstrated in favor of the active groups. In no subgroup of patients did the change in abdominal pain severity reach 30% for the Active group versus the Placebo group. However, clinical improvement was observed in many subgroups for the individual symptoms that comprise the SILSSS. Table 3 indicates that the greatest percentage improvement in IBS-SSS question scores was observed in the IBS-D subtype, particularly in women, in whom the percentages of improvement ranged from 50% to 144% in favor of the active treatment. In the diarrhea subtype, men showed less improvement in “satisfaction with bowel habits” (43%) and “interference with activity” (39%). The advantage in the IBS-C subtype was shown in “days with pain” in women (42%), and in “satisfaction with bowel habits” in both men and women (30% and 33%). LCRQQn / nznz / q / YiAi Table 3. Compendium of individual Sll-SSS questions that showed mean differences of 30% or more in favor of active treatment, mITT population Assessment Criteria Group Improvement in score, V2 to V4 Mean±SD (n) % improvement Active vs Placebo Symptoms Placebo Active Days / pain Sn-M 1.00±1.00 (3) 2.25±4.17 (8) 125 % IBS-E 9 2.14+2.97 (7) 3.03+3.74 (14) 42 % Severity IBS-D 11.31+21.27 (13) 21.33+23.82 (27) 89 % of IBS-D distension θ 16.56+15.54 (9) 24.83+24.65 (18) 50% Satisfaction with IBS-E 22 23.27±20.37 (11) 30.71±24.12 (21) 32% bowel habit IBS-C 2 24.71+25.34 (7) 32.21+24.20(14) 30% IBS-C J 20.75+9.25 (4) 27.71+25.57 (7) 33% IBS-D 22 23.00+19.77 (13) 37.82+30.95 (27) 64% IBS-D 2 21.44+29.50 (9) 37.72+35.81 (18) 76% IBS-D 2 26.50±31.10 (4) 38.00±19.58 (9) 43% All females 24.61±25.86 (18) 35.92±29.66 (40) 46 % Interfering SII-Dj¿2 16.00+21.67 (11) 32.81+26.91 (26) 105 % Group assessment criteria Improvement in score, V2 to V4 Mean+SD (n) % improvement Active vs. Placebo LCRQQn / nznz / q / Yi with life SILD 2 14.38+20.89 (8) 35.18+30.58 (17) 144 % IBS-D J 20.33±27.97 (3) 28.33±18.91 (9) 39 % IBS-M 13.00±33.20 (4) 22.80±23.64 (10) 75 % All 16.88+25.85 (17) 26.49+30.73 (40) 57 % female In many of the subgroups, the percentage in which the active treatment outperformed the placebo in the individual questions was considerably higher than our defined threshold of 30%. Quality of life overall Sil QOL scores Overall IBS-QOL scores were examined for the total population, within the IBS-C and SILD subtypes, and by gender, and by gender within each subtype. The percentage of improvement in the Active versus Placebo groups (Table 4) was comparable to the results obtained in the SIISSS, with positive responses concentrated in the IBS-D subtype and among females. A lower degree of improvement (38%) was observed in the active group among males with the IBS-D subtype. Table 4. Summary of the improvement in overall quality of life score that showed mean differences of 30% or more in favor of active treatment, mITT population Group (n) Subgroup (n) Improvement in score, visit 2 to visit 4, mean + SD (n) % improvement, Active vs Placebo Placebo Active Females and All(85) 18.44+23.15 (27) 24.01+22.39 (58) 30% Males IBS-D (37) 15.44±13.10 (11) 25.40±23.47 (26) 65% Females All (55) 11.03+18.86 (17) 21.40+20.72 (38) 94% IBS-C (20) 10.29+27.84 (7) 20.70+18.72 (13) 101% IBS-D (25) 12.78+11.61 (8) 22.40+24.78 (17) 75% Male IBS-D (12) 22.55+16.70 (3) 31.05+20.94 (9) 38% SII-QOL Quality of Life Domain Scores A therapeutic effect of the active PI over placebo was demonstrated in female subjects for overall QOL scores (Table 4) and in each of the eight domains (Table 5). The effect in the female subgroup was observed in both IBS-C and IBS-D subtypes. In the male SILD subgroup, a therapeutic effect was observed for the overall QOL score and in all four domains. Table 5. Compendium of the eight Sll-QOL domain scores, mlTT population. Changes in mean domain scores of 30% or more in favor of the active treatment are in bold. LCRQon / nznz / q / YiAi Subtypes and Genders Domain All subjects SILD SILD SILD SII-M Female Male SII-E female SII-E male SILD female SILD male Dysphoria 33.6 32.2 48.0 3.1 62.8 6.2 139.9 -10.8* 39.5 66.7 Interference with activity 42.9 27.8 78.1 8.9 208.4 -9.4* 1704.9 -28.4* 84.2 59.4 Body image 18.5 17.7* 95.5 8.6* 67.0 -9.4* 18.4 -22.2* 166.7 27.8 Health concern 45.2 19.1 112.2 46.6 80.3 19.1 32.8 11.1 131.8 19.1 Food evasion 3.5 29.9* 73.5 37.2 82.5 -33.0* -25.2* -33.7* 641.9 -31.4* Subtypes and Genres Social Reaction 36.4 65.3 29.5 15.1* 112.6 -7.3* 2331.5 -9.3* 25.0 33.6 Sexual 35.7 35.0 13.4 583.9 88.8 1.9 59.3 -9.3* 16.5 0 Relationship 29.1 2.5 68.4 51.0 69.9 4.2 91.6 -21.8* 41.7 133.1 * A negative number indicates that the improvement was greater in the Placebo group than in the Active group LCRQQn / nznz / q / Yi Adequate relief For the study population as a whole, there were no differences between the two study groups 5 with respect to the AA of Sil symptoms at visits 2, 3 and 4. A strong placebo effect was observed. Additionally, we analyzed data from each of the Sil subtypes to determine if there were differences in the SU AA within the subtypes at any study visit. No differences were found between the two study groups in any of the three Sil subtypes. Analysis of the male and female subgroups, and of the gender subgroups within each of the three Sil subtypes, yielded similar results. Stool consistency In stool consistency analysis, a positive change (improvement) indicates an increase in the BSC score in IBS-C and a decrease in the BSC score in IBS-D. Table 6 shows the percentage change, Active versus Placebo, for subgroups with changes in Active of 30% or more over Placebo. Table 6. Subgroups that showed mean differences of 30% or more for improvement in stool consistency, mITT population Subtype Criteria and Treatment Effect, % Improvement, Gender Assessment of Sil Visit 2 to Visit 4 Active vs. Placebo Placebo (n) Active (n) SILE J consistency 1.06±1.01 (4) 1.81±1.16 (8) 71% of feces BSC SILD 2J 0.93±1.37 (7) 1.78±1.42 (20) 91 % IBS-D 2 0.95±1.68 (5) 1.88±1.58 (13) 98% IBS-D J 0.88±0.18(2) 1.57±1.13 (7) 78% LCRQQn / nznz / q / Yi Mean stool consistency improved for both the placebo and active treatment groups. Changes in median stool consistency in the placebo group were typically around one BSC point (range 0.88–1.50) and approximately 1.75 BSC points in the active group (range 1.17–1.88). Percentage changes mirrored those observed in the previously presented endpoints: males and females in the active SILD subtype showed the greatest response compared to placebo. For males in the SILE subtype, there was an advantage of the active over placebo group, but this was not observed for the overall IBS-C group or for females with IBS-C. The greatest differences between treatment groups were observed in the IBS-D subtype, for both males and females.The male subgroup and the male subgroup with SILE also showed an improvement in stool consistency compared to the placebo. Frequency of bowel movements In stool frequency analysis, a positive change (improvement) indicates an increase in frequency in IBS-C and a decrease in frequency in IBS-D. In both the Placebo and Active groups, stool frequency improved in IBS-C and IBS-D subtypes, with IBS-C subjects reporting higher stool frequency during the last week of the study than during the pre-enrollment period, while IBS-D subjects reported a decrease in stool frequency during that period. Table 7 shows the IBS subtypes and subgroups in which the Active group outperformed the Placebo group in improving stool frequency by 30% or more. Table 7. Subgroups that showed mean differences of 30% or more for improvement in stool frequency, mITT population Subtype Criterion Improvement in Visit 2 score % improvement, Active rating Visit 4 vs Placebo (mean, SD) Frequency of Placebo (n) Active (n) stools / day IBS-E 0.27±0.65 (11) 0.77±0.81 (22) 185% IBS-E 9 0.29±0.49 (7) 0.57±0.76 (14) 97% IBS-E 3 0.25±0.96 (4) 1.13±0.83 (8) 352% IBS-D 0.57±2.28 (7) 1.45±1.76 (20) 154% IBS-D $ 1.20±(5) 1.62±2.02 (13) 35% SILD J -1.00± no DE (2) 1.14±1.21 (7) 214% LCRQQn / nznz / q / Yi Site-specific effects The Garden Grove clinical site had a particularly interesting subgroup of subjects: among the 16 subjects treated at Garden Grove, 12 were women with severe chronic constipation refractory to treatment. In this subgroup, the mean daily stool frequency (an important endpoint for IBS-C, according to the FDA guidance document) increased by an average of 0.25 stools / day in the Placebo group and 0.75 stools / day in the Active subgroup, a 200% increase for Active versus Placebo. Also noteworthy at this clinical site, subjects randomized to Active treatment had fewer mean stools per week at baseline than subjects in the Placebo group (0.38 stools / day vs. 0.75 stools / day), making the increase in stool frequency greater in the Active group. Table 8. Summary of improvement in daily stool frequency by Visit and treatment group, Garden Grove site with Sil of the constipation subtype, mITT population Visit 2 Visit 4 Improvement (Visit 2 to Visit 4) % of Active Placebo improvement, vs. Stools / Day Placebo Active Placebo Active Placebo Active Mean 0.75 0.38 1 1.13 0.25 0.75 200% OF 0.5 0.74 0 0.64 0.5 0.89 n 4 8 4 8 4 8 Security A total of 7 subjects reported one or more adverse events (AEs) during the study; 3 of these subjects were in the Placebo group and 4 were in the Active group. The 7 subjects reported a total of 14 AEs; all 19 were mild or moderate in severity except for severe cramps, reported by one subject in the Active group. The investigator considered four events likely to be related to the study product: dry mouth with increased thirst, increased respiration, nausea, and fatigue. All of these events were reported by one subject in the Placebo group; the dry mouth and increased thirst persisted throughout the study period but resolved the day before the subject's last study visit. No AEs were considered definitively related to the study product, and there were no serious AEs. Discussion The discovery of an effective treatment for SIL has been the goal of drug and probiotic studies in recent years. Although the subject populations in many probiotic studies have been small, several meta-analyses have been published, and certain probiotic species and strains have been shown to be more effective than others in mitigating SIL symptoms (McFarland and Dublin, 2008; Ortiz-Lucas et al., 2013). The study of SIL is complicated by the fact that patients frequently exhibit a psychological profile of anxiety and depression (Ford et al., 2014a); these psychological effects can be exacerbated by the lack of an effective treatment and the public perception of SIL as a non-serious condition. The effects of race, ethnicity, diet, and culture must also be considered in treatment evaluation (Fava et al., 2013; Hughes, 2012). In this study, the effect of the combination of L. acidophilus CL1285®, L. casei LBC80R®, and L. rhamnosus CLR2® was evaluated on IBS symptoms and quality of life in three IBS subtypes: IBS-C, IBS-D, and IBS-M. As reported in many studies, the placebo product used in this study produced improvements in IBS symptoms, reaching the level of a therapeutic effect as described in the FDA guidelines. According to these guidelines, a 30% improvement of the active product over the placebo was defined as a significant increase in therapeutic value for the study endpoint change in quality of life (overall and domains) and changes in abdominal pain, bloating, pain days, bowel habit satisfaction, and interference with life in general.The strains in the active product were effective compared to placebo in the simultaneous relief of clinical symptoms of IBS-C and IBS-D to varying degrees in both males and females. Stool frequency improved in both subtypes; stool consistency, as measured by the BSC, improved in male and female subjects with IBS-D and in male subjects with the IBS-C subtype. These endpoints are currently those recommended by regulatory agencies in the 2020s. LCRQQn / nznz / q / YiAi The United States and Europe have been used to demonstrate the efficacy of drugs in trials involving patients with IBS-C and IBS-D. These results show that these symptomatic benefits reflect parallel trends in the IBS-QOL measure developed specifically for IBS-C (Drossman et al., 2000). Female subjects, particularly those with IBS-D, responded well to the active product in terms of stool frequency and consistency, and were the most receptive in terms of symptom improvement and quality of life. While male subjects also responded well in terms of stool frequency and consistency, their response to the active product compared to the placebo was less pronounced than in the female subgroup. The symptomatic response observed in both IBS-C and IBS-D subtypes suggests that our 3 strains of lactobacilli are effective in relieving symptoms and improving quality of life in this indication. The safety profile of the product used in this study has been documented in previous clinical trials (Beausoleil et al., 2007; Gao et al., 2010; Sampalis et al., 2010) and in a quality improvement study (Maziade et al., 2015). The mechanism of action of the product under study has been demonstrated in some intestinal pathologies, but this was not investigated in the present study (Auclair et al., 2015). Interestingly, the therapeutic gains observed with our three Lactobacillus strains over placebo exceed those observed in drug studies, which are not free of significant adverse events (Cremonini et al., 2003; Kellow et al., 2003; Tack et al., 2005). Approved drugs have shown worse safety profiles than probiotic regimens, which have demonstrated an advantageous safety profile. It is worth noting that few studies have evaluated the effects of probiotics on quality of life, and of those that did, many did not find a significant improvement (Halpem et al., 1996; Kellow et al., 2003; Kim et al., 2003; Moayyedi et al., 2010; Niv et al., 2005). A few studies showed improvements in some domains (Guglielmetti et al., 2011; Kajander et al., 2008; Lorenzo-Zúñiga et al., 2014; O'Mahony et al., 2005), but to our knowledge, no effect has previously been documented in the "interference with activity" domain. O'Mahony et al. found lower IBS-QOL scores for L. salivarius ssp. and B. infantis for most domains (O'Mahony et al., 2005). This study provides evidence of the therapeutic effects on specific subtypes and subgroups of SIL, which were consistently observed across various endpoints: stool frequency and consistency, quality of life, improvement in bloating severity, days with pain, and satisfaction with bowel habits. Our findings are consistent with other studies conducted with probiotics and medications (Somberg, 2012). A low incidence of adverse events (AEs) has been observed in previous studies conducted with the investigational product. This protocol was based on previous clinical studies conducted in Canada and the United States, using the optimal dose of the product. The investigational product was also previously evaluated in adults for antibiotic-associated diarrhea and C. difficile prevention, demonstrating a significant reduction in the risk of diarrhea during a C. difficile outbreak in China (Gao et al., 2010). In the last decade of clinical research involving the investigational product, there have been no serious adverse events (SAEs) related to the investigational product in any of the clinical trials (Beausoleil et al., 2007; Gao et al., 2010; Maziade et al., 2015; Sampalis et al., 2010). Conclusions The specific combination of live bacteria used in this study produced results that varied between genera and subtypes. However, it had a clear positive impact on stool consistency and frequency, quality of life, and SIL symptoms in both sexes, without serious adverse events. These findings present a promising therapeutic option for individuals with SIL. References Aagaard, K., Petrosino, J., Keitel, W., Watson, M., Katancik, L, García, N., Patel, S., Cutting, M., Madden, T., Hamilton, H., Harris, E., Gevers, D., Simone, G., Mclnnes, P. and Versalovic, L, 2013. The Human Microbiome Project strategy for comprehensive sampling of the human microbiome and why it matters. Federation of American Societies for Experimental Biology Magazine 27: 10121022. Auclair, L, Frappier, M. y Millette, M., 2015. Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, y Lactobacillus rhamnosus CLR2 (Bio-K+): Characterization, Manufacture, Mechanisms of Action, and Quality Control of a Specific Probiotic Combination for Primary Prevention of Clostridium difficile Infection. Clinical Infectious Diseases 60 Suplemento 2: S135143. Beausoleil, M., Fortier, N., Guénette, S., L'ecuyer, A., Savoie, M., Franco, M., Lachaine, J. y Weiss, K., 2007. Effect of a fermented milk combining Lactobacillus acidophilus C11285 and Lactobacillus casei in the prevention of antibiotic-associated diarrhea: a randomized, double-blind, placebo-controlled trial. Canadian Journal of Gastroenterol 21: 732-736. Camilleri, M., Mayer, E.A., Drossman, D.A., Heath, A., Dukes, G.E., McSorley, D., Kong, S., Mangel, A.W. y Northcutt, A.R., 1999. Improvement in pain and bowel function in female irritable bowel patients with alosetron, a 5-HT3 receptor antagonist. Alimentary Pharmacology & Therapeutics 13: 1149-1159. Coffin, B., Dapoigny, M., Cloarec, D., Comet, D. y Dyard, F., 2004. Relationship between severity of symptoms and quality of life in 858 patients with irritable bowel syndrome. Gastroentérologie Clinique et Biologique 28: 11-15. Cremonini, F., Delgado-Aros, S. y Camilleri, M., 2003. Efficacy of alosetron in irritable bowel syndrome: a meta-analysis of randomized controlled triáis. Neurogastroenterology & Motility 15: 79-86. Drossman, D.A., Patrick, D.L., Whitehead, W.E., Toner, B.B., Diamant, N.E., Hu, Y., Jia, H. y Bangdiwala, S.I., 2000. Further validation of the IBS-QOL: a disease-specific quality-of-life questionnaire. American Journal of Gastroenteroloy 95: 999-1007. Ducrotté, P., Sawant, P. y Jayanthi, V., 2012. Clinical trial: Lactobacillus plantarum 299v (DSM 9843) improves symptoms of irritable bowel syndrome. World Journal of Gastroenterology 18: 4012-4018. Fava, F., Gitau, R., Griffin, B.A., Gibson, G.R., Tuohy, K.M. y Lovegrove, J.A., 2013. The type and quantity of dietary fat and carbohydrate alter faecal microbiome and short-chain fatty acid excretion in a metabolic syndrome 'at-risk' population. International Journal of Obesity (London) 37: 216-223. Ford, A.C. y Moayyedi, P., 2010. Meta-analysis: factors affecting placebo response rate in the irritable bowel syndrome. Alimentary Pharmacology & Therapeutics 32: 144-158. Ford, A.C., Quigley, E.M., Lacy, B.E., Lembo, A.J., Saito, Y.A., Schiller, L.R., Soffer, E.E., Spiegel, B.M. y Moayyedi, P., 2014a. Effect of antidepressants and psychological therapies, including hypnotherapy, in irritable bowel syndrome: systematic review and meta-analysis. American Journal of Gastroenterology 109: 1350-1365; cuestionario 1366. Ford, A.C., Quigley, E.M., Lacy, B.E., Lembo, A.J., Saito, Y.A., Schiller, L.R., Soffer, E.E., Spiegel, B.M. y Moayyedi, P., 2014b. Efficacy of prebiotics, probiotics, and synbiotics in irritable bowel syndrome and chronic idiopathic constipation: systematic review and meta-analysis. American Journal of Gastroenterology 109: 1547-1561; cuestionario 1546, 1562. Foxx-Orenstein, A., 2006. IBS—revisión y novedades. Medicina General de Medscape 8: 20. Gao, X.W., Mubasher, M., Fang, C.Y., Reifer, C. y Miller, L.E., 2010. Dose-response efficacy of a 23 LCRQon / nznz / q / YiAi proprietary probiotic formula of Lactobacillus acidophilus CL1285 and Lactobacillus casei LBC80R for antibiotic-associated diarrhea and Clostridium difficile-associated diarrhea prophylaxis in adult patients. American Journal of Gastroenterology 105: 1636-1641. Guglielmetti, S., Mora, D., Gschwender, M. y Popp, K., 2011. Randomised clinical trial: Bifidobacterium bifidum MIMBb75 significantly alleviates irritable bowel syndrome and improves quality of life—a double-blind, placebo-controlled study. Alimentary Pharmacology & Therapeutics 33: 1123-1132. Halpern, G.M., Prindiville, T., Blankenburg, M., Hsia, T. y Gershwin, M.E., 1996. Treatment of irritable bowel syndrome with Lacteol Fort: a randomized, double-blind, cross-over trial. American Journal of Gastroenterology 91: 1579-1585. Hoveyda, N., Heneghan, C., Mahtani, K.R., Perera, R., Roberts, N. y Glasziou, P., 2009. A systematic review and meta-analysis: probiotics in the treatment of irritable bowel syndrome. BMC Gastroenterology 9: 15. Hughes, V., 2012. Microbiome: Cultural differences. Nature 492: S14-15. Kajander, K., Myllyluoma, E., Rajilic-Stojanovic, M., Kyronpalo, S., Rasmussen, M., Járvenpáa, S., Zoetendal, E.G., de Vos, W.M., Vapaatalo, H. y Korpela, R., 2008. Clinical trial: multispecies probiotic supplementation alleviates the symptoms of irritable bowel syndrome and stabilizes intestinal microbiota. Alimentary Pharmacology & Therapeutics 27: 48-57. Kellow, J., Lee, O.Y., Chang, F.Y., Thongsawat, S., Mazlam, M.Z., Yuen, H., Gwee, K.A., Bak, Y.T., Jones, J. y Wagner, A., 2003. An Asia-Pacific, double blind, placebo controlled, randomised study to evalúate the efficacy, safety, and tolerability of tegaserod in patients with irritable bowel syndrome. Gut 52: 671-676. Kim, H.J., Camilleri, M., McKinzie, S., Lempke, M.B., Burton, D.D., Thomforde, G.M. y Zinsmeister, A.R., 2003. A randomized controlled trial of a probiotic, VSL#3, on gut transit and symptoms in diarrhoea-predominant irritable bowel syndrome. Alimentary Pharmacology & Therapeutics 17: 895-904. Lorenzo-Zúñiga, V., Llop, E., Suárez, C., Alvarez, B., Abren, L., Espadaler, J. y Serra, J., 2014. 1.31, a new combination of probiotics, improves irritable bowel syndrome-related quality of life. World Journal of Gastroenterology 20: 8709-8716. Lyra, A., Hillilá, M., Huttunen, T., Mánnikkó, S., Taalikka, M., Tennilá, J., Tarpila, A., Lahtinen, S., Ouwehand, AC y Veijola, L., 2016. Irritable bowel syndrome symptom severity improves equally with probiotic and placebo. World Journal of Gastroenterology 22: 10631-10642. Maziade, P.J., Pereira, P. y Goldstein, E.J., 2015. A Decade of Experience in Primary Prevention of Clostridium difficile Infection at a Community Hospital Using the Probiotic Combination Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, and Lactobacillus rhamnosus CLR2 (BÍO-K+). Clinical Infectious Diseases 60 Suplemento 2: S144-147. McFarland, L.V. y Dublin, S., 2008. Meta-analysis of probiotics for the treatment of irritable bowel syndrome. World Journal of Gastroenterology 14: 2650-2661. Moayyedi, P., Ford, A.C., Talley, N.J., Cremonini, F., Foxx-Orenstein, A.E., Brandt, L.J. y Quigley, E.M., 2010. The efficacy of probiotics in the treatment of irritable bowel syndrome: a systematic review. Gut 59: 325-332. Niv, E., Naftali, T., Hallak, R. y Vaisman, N., 2005. The efficacy of Lactobacillus reuteri ATCC 55730 in the treatment of patients with irritable bowel syndrome-a double blind, placebocontrolled, randomized study. Clinical Nutrition 24: 925-931. O'Mahony, L., McCarthy, J., Kelly, P., Hurley, G., Luo, F., Chen, K., O'Sullivan, G.C., Kiely, B., Collins, J.K., Shanahan, F. y Quigley, E.M., 2005. Lactobacillus and Bifidobacterium in irritable bowel syndrome: symptom responsos and relationship to cytokine profiles. Gastroenterology 128: 541-551. Ortiz-Lucas, M., Tobías, A., Saz, P. y Sebastian, J.J., 2013. Effect of probiotic species on irritable bowel syndrome symptoms: A bring up to date meta-analysis. Revista Española de Enfermedades Digestivas 105: 19-36. Peery, A.F., Dellon, E.S., Lund, J., Crockett, S.D., McGowan, C.E., Bulsiewicz, W.J., Gangarosa, L.M., Thiny, M.T., Stizenberg, K., Morgan, D.R., Ringel, Y., Kim, H.P., Dibonaventura, M.D., CarroU, C.F., Alien, J.K., Cook, S.F., Sandler, R.S., Kappelman, M.D. y Shaheen, N.J., 2012. Burden of gastrointestinal disease in the United States: 2012 update. Gastroenterology 143: 11791187 ell71-1173. Sampalis, J., Psaradellis, E. y Rampakakis, E., 2010. Efficacy of ΒΙΟ K+ CL1285 in the reduction of antibiotic-associated diarrhea - a placebo controlled double-blind randomized, multi-center study. Archives of Medical Science 6: 56-64. Shih, D.Q. y Kwan, L.Y., 2007. All Roads Lead to Rome: Update on Rome III Criteria and New Treatment Options. Gastroenterology Reporte 1: 56-65. Somberg, J.C., 2012. The human microbiome and therapeutics. American Journal of Therapeutics 19: 247 Tack, J., Müller-Lissner, S., Bytzer, P., Corinaldesi, R., Chang, L., Viegas, A., Schnekenbuehl, S., 25 LCRQQn / nznz / q / YiAi Dunger-Baldauf, C. y Rueegg, P., 2005. A randomised controlled trial assessing the efficacy and safety of repeated tegaserod therapy in women with irritable bowel syndrome with constipation. Gut 54: 1707-1713. Whelan, K. y Myers, C.E., 2010. Safety of probiotics in patients receiving nutritional support: a systematic review of case reports, randomized controlled triáis, and nonrandomized triáis. American Journal of Clinical Nutrition 91: 687-703. Yoon, H., Park, Y.S., Lee, D.H., Seo, J.G., Shin, C.M. y Kim, N., 2015. Effect of administering a multi-species probiotic mixture on the changes in fecal microbiota and symptoms of irritable bowel syndrome: a randomized, double-blind, placebo-controlled trial. Journal of Clinical Biochemistry and Nutrition 57: 129-134. The headings are included in this description for reference and to aid in locating certain sections. These headings are not intended to limit the scope of the concepts described therein, and these concepts may have applicability in other sections throughout the specification. Therefore, the present invention is not intended to be limited to the embodiments shown in this description, but rather to have the broadest scope consistent with the novel principles and features described herein. The singular forms “a”, “an”, and “the” include the corresponding plural references unless the context clearly dictates otherwise. Thus, for example, a reference to a compound includes one or more such compounds, and a reference to the method includes references to equivalent steps and methods known to those skilled in the art that could be modified or substituted for the methods described herein. Unless otherwise stated, all numbers expressing quantities of ingredients, reaction conditions, concentrations, properties, etc., used in the description and claims shall be understood to be modified in all cases by the term "approximately." At a minimum, each numerical parameter should be interpreted in light of the number of significant digits reported and by applying ordinary rounding techniques. Consequently, unless otherwise stated, the numerical parameters set forth in this description and accompanying claims are approximations that may vary depending on the desired properties. Although the numerical ranges and parameters that describe the broadest scope of the formulations are approximations, the numerical values ​​given in specific examples are reported as precisely as possible.Any numerical value, however, inherently contains certain errors that result from the 26. LCRQon / nznz / q / YiAi variations in experiments, test measurements, statistical analyses, and others. It is understood that the examples and modalities described in the present description are for illustrative purposes only and that various modifications or changes in light of them will be suggested by those skilled in the art and should be included within the present invention and the scope of the appended claims. LCRQQn / nznz / q / YiAi

Claims

1. A method for relieving a gastrointestinal disorder in a subject in need, comprising administering to said subject a combination of live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus, wherein said gastrointestinal disorder is selected from the group consisting of irritable bowel syndrome (IBS), abdominal pain, abdominal distension, and constipation.

2. A method for improving the quality of life of a subject suffering from irritable bowel syndrome (IBS), comprising administering to said subject a combination of live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus, wherein said administration provides said subject with at least one benefit selected from the group consisting of: satisfaction with bowel habits, minimal interference of IBS with normal activity, improved body image, reduced food avoidance, increased social reaction, reduced sexual dysfunction, and improved relationships.

3. The method of claim 1 or 2, wherein said Lactobacillus acidophilus is Lactobacillus acidophilus CL1285® deposited in the CNCM, wherein said Lactobacillus casei is Lactobacillus casei LBC80R® deposited in the CNCM, and wherein said Lactobacillus rhamnosus is Lactobacillus rhamnosus CLR2® deposited in the CNCM.

4. The method of any of claims 1 to 3, wherein said administration comprises the administration of a nutritionally acceptable composition comprising said combination of Lactobacilli.

5. The method of any of claims 1 to 4, wherein said administration comprises administering at least 10 to 200 trillion of said Lactobacillus combination.

6. The method of any of claims 1 to 4, wherein said combination of live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus comprises approximately 1-10% L. acidophilus, 70-90% L. casei, and approximately 5-20% L. rhamnosus of the colony-forming units (CFU) of the combination.

7. The method of any of claims 1 to 6, wherein said administration comprises administering said combination of Lactobacilli at least once a day.

8. The method of any of claims 1 to 7, wherein said administration comprises administering a capsule comprising said combination of Lactobacilli.

9. The method of any of claims 1 to 8, wherein said administration comprises administering a ferment, said ferment comprising fermented proteins in addition to said combination of Lactobacilli.

10. The method of claim 9, wherein said fermented proteins are selected from the group consisting of fermented soybean proteins, fermented milk proteins, fermented rice proteins, fermented pea proteins, fermented hemp proteins, fermented almond proteins, and fermented insect proteins.

11. A method for the relief of irritable bowel syndrome (IBS) in a human subject in need, comprising administering to said subject a combination of live Lactobacillus acidophilus CL1285®, live Lactobacillus casei LBC80R® and live Lactobacillus rhamnosus CLR2®.

12. The method of claim 11, wherein said IBS comprises at least one of IBS with predominant constipation (IBS-C), IBS with predominant diarrhea (IBS-D), and IBS with mixed bowel patterns (IBS-M).

13. The method of claim 12, wherein said method provides simultaneous relief of multiple symptoms of Sil.

14. The method of claim 13, wherein said symptoms are selected from the group consisting of abdominal pain, abdominal distension, and constipation.

15. The method of any of claims 11 to 14, wherein said administration comprises the administration of a nutritionally acceptable composition comprising said LCRQQn / nznz / q / YiAi combination of Lactobacilli.

16. The method of any of claims 11 to 15, wherein said administration comprises administering at least 10 to 200 billion of said Lactobacillus combination.

17. The method of any of claims 11 to 16, wherein said combination of live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus comprises approximately 1-10% L. acidophilus, 70-90% L. casei, and approximately 5-20% L. rhamnosus of the colony-forming units (CFU) of the combination.

18. The method of any of claims 11 to 17, wherein said administration comprises administering said combination of Lactobacilli at least once a day.

19. The method of any of claims 11 to 17, wherein said administration comprises administering a capsule comprising said combination of Lactobacilli.

20. The method of any of claims 11 to 19, wherein said administration comprises administering a ferment, said ferment comprising fermented proteins in addition to said combination of Lactobacilli.

21. The method of claim 20, wherein said fermented proteins are selected from the group consisting of fermented soybean proteins, fermented milk proteins, fermented rice proteins, fermented pea proteins, fermented hemp proteins, fermented almond proteins, and fermented insect proteins.

22. An alternative method to drug therapy for the prevention and / or treatment of irritable bowel syndrome (IBS), comprising: (i) identifying a human subject who requires drug therapy for the prevention and / or treatment of IBS; and (ii) administering to said human subject a nutritionally acceptable composition comprising a combination of live microorganisms comprising live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus in addition to or as a replacement for said drug therapy. LCRQon / nznz / q / YiAi 23. The alternative method according to claim 22, wherein said Lactobacillus acidophilus is Lactobacillus acidophilus CL1285® deposited in the CNCM, wherein said Lactobacillus casei is Lactobacillus casei LBC80R® deposited in the CNCM, and wherein said Lactobacillus rhamnosus is Lactobacillus rhamnosus CLR2® deposited in the CNCM.

24. The alternative method according to claim 22 or 23, wherein said administration comprises administering said nutritionally acceptable composition at least once a day.

25. The alternative method of any of claims 22 to 24, wherein said administration comprises administering a capsule comprising said nutritionally acceptable composition.

26. The alternative method of any of claims 22 to 25, wherein said nutritionally acceptable composition comprises at least 10 to 200 trillion of said Lactobacillus combination.

27. The method of any of claims 22 to 26, wherein said combination of live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus comprises approximately 1-10% L. acidophilus, 70-90% L. casei, and approximately 5-20% L. rhamnosus of the colony-forming units (CFU) of the combination.

28. The alternative method of any of claims 22 to 27, wherein said nutritionally acceptable composition takes the form of a ferment comprising fermented proteins in addition to said combination of Lactobacilli.

29. The alternative method according to claim 28, wherein said fermented proteins are selected from the group consisting of fermented soy protein, fermented milk protein, fermented rice protein, fermented pea protein, and fermented hemp protein.

30. The method of any of claims 22 to 29, wherein said administration LCRQQn / nznz / q / YiAi provides said subject with at least one benefit selected from the group consisting of: satisfaction with bowel habits, minimal interference of Sil with normal activity, improved body image, reduced food avoidance, increased social reaction, reduced sexual dysfunction, and improved relationships.

31. A composition for the relief of a gastrointestinal disorder in a subject in need, the composition comprising a combination of live Lactobacillus acidophilus CL1285®, live Lactobacillus casei LBC80R®, live Lactobacillus rhamnosus CLR2®, wherein said gastrointestinal disorder is selected from the group consisting of irritable bowel syndrome (IBS), abdominal pain, days of pain, distension, stool consistency, and stool frequency.

32. A composition for the relief of irritable bowel syndrome (IBS) in a subject in need, the composition comprising a combination of live Lactobacillus acidophilus CL1285®, live Lactobacillus casei LBC80R® and live Lactobacillus rhamnosus CLR2®.

33. The composition of claims 31 or 32, wherein said composition comprises fermented proteins in addition to said combination of Lactobacilli.

34. The composition according to claim 33, wherein said fermented proteins are selected from the group consisting of fermented soy protein, fermented milk protein, fermented rice protein, fermented pea protein, fermented hemp protein, fermented almond protein, and fermented insect protein.

35. The composition of any of claims 31 to 34, wherein said composition comprises at least 10 to 200 trillion of said combination of Lactobacillus.

36. The composition of any of claims 31 to 35, wherein said composition comprises approximately 1-10% of L. acidophilus, 70-90% of L. casei and approximately 5-20% of L. rhamnosus of the colony-forming units (CFU) of the combination.

37. The composition of any of claims 31 to 36, wherein said composition is a nutritionally acceptable composition.

38. The use of a combination of live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus, for the relief of a gastrointestinal disorder in a subject in need, wherein said gastrointestinal disorder is selected from the group consisting of irritable bowel syndrome (IBS), abdominal pain, abdominal distension, and constipation.

39. The use according to claim 38 wherein said Lactobacillus acidophilus is Lactobacillus acidophilus CL1285® stock No. CNCM 1-4099, wherein said Lactobacillus casei is Lactobacillus casei LBC80R® stock No. CNCM 1-3989, and wherein said Lactobacillus rhamnosus is Lactobacillus rhamnosus CLR2® stock No. CNCM 1-3990.

40. Use according to claim 38 or 39, wherein said administration provides said subject with at least one benefit selected from the group consisting of: satisfaction with bowel habits, minimal interference of the Sil with normal activity, improved body image, reduced food avoidance, increased social reaction, reduced sexual dysfunction, and improved relationships.

41. The use of a combination of live Lactobacillus acidophilus CL1285®, live Lactobacillus casei LBC80R® and live Lactobacillus rhamnosus CLR2®, for the prevention, treatment and / or relief of irritable bowel syndrome (IBS) in a subject who needs it.

42. The use of any of claims 38 to 41, wherein said combination of live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus comprises approximately 1-10% L. acidophilus, 70-90% L. casei, and approximately 5-20% L. rhamnosus of the colony-forming units (CFU) of the combination.

43. The use of any of claims 38 to 42, wherein said live Lactobacillus acidophilus, live Lactobacillus casei, and live Lactobacillus rhamnosus are present in a composition comprising fermented proteins.

44. For the use according to claim 43, said fermented proteins are selected from the LCRQQn / nznz / q / YiAi group consisting of fermented soy protein, fermented milk protein, fermented rice protein, fermented pea protein, fermented hemp protein, fermented almond protein, and fermented insect protein. 5 45. For the use of any of claims 38 to 44, wherein said subject ingests at least 10 to 200 trillion of said Lactobacillus combination once a day.