Trial 132 Clinical results of treatment with the lactic yeast Kluyveromyces B0399 for Irritable Bowel Sindrome. This study, based on the gathering of symtomatological data before and after the treatment, demonstrates that the prolonged use of Kluyveromyces marxianus fragilis B0399 significantly improves the clinical picture and above all improves the quality of life of these patients.
Dr. Sandro Andreoli*
Dr. Paola Lovrovich (statistical analysis)
*Doctor in Medicine and Surgery; Specialist in Diseases of the Degistive System
Endoscope specialist at the S.Camillo Hospital, Treviso
Gastroenterology consultant at the Città della Salute, Torreano di Martignacco (Ud)
Gastroenterology consultant at the Salus Alpe Adria Pagnacco (Ud)
Gastroenterology consultant at the Clinica S. Eufemia Grado (Go).
Private Studio in Via Duino 1, 33100 Udine
0-SUMMARY:
Irritable Bowel Syndrome is provoked by a multitude of causes (physiological, psychological, environmental and behavioral) therefore no effective specific therapy exists, but rather multiple therapies that are aimed at the causes which are subjectively most important. This study, based on the gathering of symtomatological data before and after the treatment, demonstrates that the prolonged use of Kluyveromyces marxianus fragilis B0399 significantly improves the clinical picture and above all improves the quality of life of these patients.
1-INTRODUCTION:
Since 1998, the use of the lactic yeast Kluyveromyces marxianus fragilis B0399 is applied as a support aimed at significantly limiting the symptomology of the irritable colon, give significant proof of effectiveness, so much so as to encourage further study.
The objective of this study was to verify if the cyclical use of the lactic yeast Kluyveromyces marxianus fragilis B0399 improves the symptomology in patients affected by Irritable Bowel Syndrome (IBS). The utilization of subjects affected by IBS is recognized internationally (as illustrated in “The EFSA Journal (2008)853,1-15) as experimentation aimed at proving the effectiveness in reducing “intestinal discomfort”.
Numerous conferences have been dedicated to IBS with the objective of establishing a definition and the symptomology .
International groups of experts, brought together for the theme of Functional Gastrointestinal Disorders, have elaborated diagnostic criteria, known by the name of Roma I, II and III and have defined IBS as being “a group of functional intestinal disorders, in which abdominal pain is associated with defecation or the modification of the routine of the intestine or the subjective perception of altered defecation.” IBS is also defined as being a dysfunction and can include disorders known as: irritable colon, spastic colitis or simply colitis.
Abdominal pain or discomfort (disturbing sensation) recurring for at least three days a month in the last 3 months, associated with at least two of the following symptoms:
Abdominal pain can manifest itself as a cramp, predominantly during the day, alleviated by defecation and exacerbated by stress and the intake of food. This symptomology often conditions the quality of life and daily routine. The altered frequency of evacuations is expressed with a prevalence of constipation as opposed to diarrhea, especially among women. Men, instead, have a prevalence of diarrhea.
IBS is not a disease but a dysfunction, caused by an exaggerated synergy between the Central Nervous System and the Enteric Nervous System. It doesn’t have a well-defined etiologic factor; it is considered a “bio-psychosocial” disorder, caused by Physiological, psychological, environmental and behavioral factors.
IBS frequently has cyclical occurrences, with periods of well-being alternating with painful attacks provoked by emotional tension, fatigue, stress, diet. These painful crises create further tension in the patient which can cause a worsening of the symptoms. Genetic factors combined with psychosocial factors determine an increase of intestinal inflammatory cells, with an increase and activation of the mast cells, which cause colic hypersensitivity and unmobility, with bloating, alteration of transit time, meteorism, hypertonia of the sigmoid flexure, appearance of fecal mucous. Colonoscopy is often macroscopically negative but the histology highlights the microscopic colitis.
According to epidemiological data, IBS is present in 30% of the Italian population with a ratio woman/man between 2:1 and 4:1. This tendency is also found in our clinical experience with a ratio of 7 out of ten. This might indicate a greater sensitivity and knowledge on the part of the women.
Diagnosis
The diagnosis of IBS is based on an accurate evaluation of clinical symptoms and on the exclusion of other pathologies. A method had recently been set up consistent measuring fecal Calprotectina. In this way, the organic disease (colitis) is distinguished from the alteration of the functional state and is therefore considered to be the best means to confirm the diagnosis of IBS, excluding other pathologies such as intestinal inflammatory disease (IBS), poor absorption, tumors, infections (Histolytic entamoeba, Yersinia, Campylobacter jejuni, Giardia, etc), diets too rich in sorbitol, fructose, fiber, lactose, the use of laxatives or drugs rich in magnesium.
2-MATERIALS AND METHODS
2.1- The active ingredient
The active ingredient is the lactic yeast Kluyveromyces marxianus fragilis B0399, produced by Turval Laboratories of Udine. The product in capsule is already in commerce in pharmacies and has been notarized by the Ministry of Health.
2.2-Selection of the group to be examined.
Fifty people who came under our observation during the past two years were selected; fifteen were men with an average age of 33 (20-62) and 35 women with an average age of 31(22-55). None of the subjects presented other pathologies, gastroenterological or otherwise. None were using any type of drug. They had symptoms which indicated Irritable Bowel Syndrome. To exclude ulterior pathologies, haematic-chemical exams such as bood count, Ves, Hepatic profile, amilasy test,total IgE, sideremia, search of Hb in huaman feces (three sample),fecal parasitological exam, feces culture exam (in patients with diarrhea), fecal Calprotectina dosage.
Patients over 50 years old and those who had a family history with many cases of colic polyps underwent a total colonoscopy (11). Three had colic polyps which were removed endoscopically (slight and mediumdysplasia ).
The 50 patients whose absence of pathological alterations worthy of mention was verified were asked to answered a series of question asked by the physician that summarize the diagnostic criteria of Roma (I, II and III). This questionnaire, defined as pre-administration, refers back to the study conducted at the University Hospital of Manchester (Francis at all, 1996) recommended in “Design of Treatment Trials for Functional Gastrointestinal Disorders” (Irvine at all., 2006), considered the reference guide of the EFSA.
The people who resulted in being suitable and were part of the program were adequately instructed on the lifestyle to follow, foods and dietary mistakes to avoid (coffee, carbonated drinks, alcohol, hot spices, legumes, cocoa, milk and dairy products, oily dry fruit, etc.)
2.3-Daily dosage
The experimentation in exam consisted in the administration of one capsuleevery twelve hours for a month and a maintenance period with one capsulebefore breakfast for two months, as normally recommended by the producing company and by doctors (pediatricians, gastroenterologists, gynecologists, family practitioners, etc.) as a dietary supplement.
Based on the information provided by the producing company, one capsule contains not less than 10/20 million live cells of lactic yeast .
2.4-Analysis of the questionnaires
At the end of the period of administration of the product, the patients were visited again and a post-administration questionnaire was collected for a verification of the symptoms.
From the analysis of the answers obtained in the questionnaire before the administration and the one after administration, the evidence of an improvement in IBS, or lack thereof, was examined.
2.5-Diagnostic criteria.
Before the experimentation, diagnostic criteria for the determining of cases of IBS and their characteristics was selected that go by the name of Roma (I,II and III). This criteria helped to determine the suitability of the people chosen for the experimentation.
|
Questionnaire question pre-administration
|
Answers that satisfy the diagnostic criteria
|
Answers that don’t satisfy the diagnostic criteria
|
|
1
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3 / 4 / 5 / 6
|
0 / 1 / 2
|
|
2
|
0 / 2
|
1
|
|
|
Suitable patient
|
Unsuitable patient
|
|
Questionnaire question pre-administration
|
Answers that satisfy the diagnostic criteria
|
Answers that don’t satisfy the diagnostic criteria
|
|
3
|
> 0
|
0
|
|
4
|
> 0
|
0
|
|
5
|
> 0
|
0
|
|
6
|
> 0
|
0
|
|
7
|
> 0
|
0
|
|
8
|
> 0
|
0
|
|
Subtype of IBS
|
Question 9 (pre-adm.)
|
Question 10 (pre-adm.)
|
|
Constipation
|
>0
|
0
|
|
Diarrhea
|
0
|
>0
|
|
Mixed
|
>0
|
>0
|
|
Unclassifiable
|
0
|
0
|
|
Question of pre-administration test.
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Question of post-administration test.
|
|
1
|
I
|
|
2
|
II
|
|
3
|
|
|
4
|
III
|
|
5
|
IV
|
|
6
|
IV
|
|
7
|
V
|
|
8
|
VI
|
|
9
|
VII
|
|
10
|
VIII
|
|
11
|
IX
|
|
12
|
XII
|
|
|
III
|
|
|
X
|
|
|
XI
|
3-STATISTICAL ANALYSIS :
3.1-Statistical analysis of the Chi-square Yates/Pearson
For the analysis of the data obtained from the answers of the questionnaires both pre and post-administration the analytic procedure of the Chi-squareof Yates and the Chi-squarePearson (with Vassar Stats: Statistical Computation Web Site) was selected, with the aim of verifying if their differences are due to chance or not. If the differences are not due to chance, they would be considered “statistically significant.”
Chi-square: Initially, whatever the existing differences between the two series of data to compare, the “zero hypothesis” is advanced. The zero hypothesis (or null hypothesis) simply affirms that the difference observed – of whatever entity – is due to chance. This hypothesis can either be accepted or rejected on the basis of the results of the statistical test.
If on the basis of this hypothesis the calculated value of χ² is greater than a certain critical value, we’ll have to conclude that the frequencies observed differ significantly from the frequencies expected and we will have to refuse the H0 at the corresponding level of significance. Otherwise, we will have to accept it, or at least not refute it. This procedure is called chi-squareof the hypothesis.
To proceed in determining the difference between a “healthy” state of a person and one affected by IBS, the answers to the questions were grouped, according the description shown below.
First question: following the diagnostic criteria described in paragraph 2.5, the “healthy” people reply with the options 0-1-2 while those affected by IBS reply with options 3-4-5-6.
Fourth question: following the diagnostic criteria described in paragraph 2.5, the “healthy” people reply with the options 0-1 while those affected by IBS reply with options 2-3-4.
Seventh, eighth, ninth, tenth and eleventh question: following the diagnostic criteria described in paragraph 2.5, the “healthy” people reply with the options 0-1 while those affected by IBS reply with options 2-3-4.
Furthermore, the division of these two cases identifies in answers 0-1 the sporadic nature of the event and in answers 2-3-4 indicate association with pain and elevated frequency.
5-Initial questionnaire (pre-administration):
7-RESULTS:
7.1-comparison between the pre-administration questionnaire filled out at the doctor’s office and the one mailed from home by the patient.
The comparison between the initial forms collected by the doctor and those collected from the patients at both time 0 and at the end (after three months of the administration of Kluyveromyces marxianus fraglis B0399) showed an almost total correlation between the forms collected by the doctor and those collected from the patients: the discordance is minimal (2% of the answers).
The interpretation of the answers to the questionnaire is shown in the following paragraphs:
|
Answer
|
Question 1
(pre-administration)
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Question I
(post-administration)
|
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0 (never)
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0
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2
|
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1 (< 1 day a month)
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0
|
10
|
|
2 ( 1 day a month)
|
0
|
25
|
|
3 (2-3 days a month)
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4
|
8
|
|
4 (1 day a week)
|
12
|
5
|
|
5 ( > 1 day a week)
|
29
|
0
|
|
6 (every day)
|
5
|
0
|
|
|
Chi-square
|
|
|
Phi
|
Yates
|
Pearson
|
|
+0,77
|
55,6
|
58,73
|
|
P
|
<0,001
|
<0,001
|
|
Answer
|
Question 2
(pre-administration)
|
Question II
(post-administration)
|
|
0
|
31
|
31
|
|
1
|
0
|
0
|
|
2
|
4
|
4
|
|
Answer
|
Question 3
(pre-administration)
|
|
0
|
0
|
|
1
|
50
|
|
Answer
|
Question 4
(pre-administration)
|
Question III
(post-administration)
|
|
0 (never / rarely)
|
0
|
2
|
|
1 (sometimes)
|
12
|
20
|
|
2 (often)
|
6
|
18
|
|
3 (most of the time)
|
28
|
6
|
|
4 (always)
|
4
|
1
|
|
|
Chi-square
|
|
|
Phi
|
Yates
|
Pearson
|
|
+0,24
|
4,58
|
5,54
|
|
P
|
0,032347
|
0,018587
|
|
Answer
|
Question 5
(pre-administration)
|
Question 6
(pre-administration)
|
|
Answer
|
Question IV
(post-administration)
|
|
0 (never/rarely)
|
14
|
4
|
|
0 (like in the past)
|
0
|
|
1 (sometimes)
|
12
|
14
|
|
1 (more frequent)
|
3
|
|
2 (often)
|
10
|
6
|
|
2 (less frequent)
|
15
|
|
3 (most of the time)
|
8
|
2
|
|
3 (normalized)
|
32
|
|
4 (always)
|
6
|
1
|
|
|
|
|
Answer
|
Question 7
(pre-administration)
|
Question V
(post-administration)
|
|
0 (never / rarely)
|
14
|
18
|
|
1 (sometimes)
|
10
|
12
|
|
2 (often)
|
12
|
6
|
|
3 (most of the time)
|
8
|
0
|
|
4 (always)
|
6
|
0
|
|
|
Chi-square
|
|
|
Phi
|
Yates
|
Pearson
|
|
+0,36
|
9,72
|
11,18
|
|
P
|
0,001823
|
0,000827
|
|
Answer
|
Question 8
(pre-administration)
|
question VI
(post-administration)
|
|
0 (never / rarely)
|
2
|
10
|
|
1 (sometimes)
|
13
|
8
|
|
2 (often)
|
5
|
3
|
|
3 (most of the time)
|
2
|
2
|
|
4 (always)
|
1
|
0
|
|
|
Chi-square
|
|
|
Phi
|
Yates
|
Pearson
|
|
+0,14
|
0,43
|
0,97
|
|
P
|
0,511989
|
0,32468
|
|
Answer
|
Question 9
(pre-administration)
|
Question VII
(post-administration)
|
|
0 (never / rarely)
|
4
|
12
|
|
1 (sometimes)
|
6
|
8
|
|
2 (often)
|
9
|
3
|
|
3 (most of the time)
|
5
|
0
|
|
4 (always)
|
3
|
0
|
|
|
Chi-square
|
|
|
Phi
|
Yates
|
Pearson
|
|
+0,51
|
10,9
|
12,9
|
|
P
|
0,000962
|
0,000329
|
|
Answer
|
Question 10
(pre-administration)
|
Question VIII
(post-administration)
|
|
0 (never/ rarely)
|
14
|
26
|
|
1 (sometimes)
|
12
|
10
|
|
2 (often)
|
14
|
0
|
|
3 (most of the time)
|
8
|
0
|
|
4 (always)
|
2
|
0
|
|
|
Chi-square
|
|
|
Phi
|
Yates
|
Pearson
|
|
+0,53
|
21,64
|
23,97
|
|
P
|
<0,001
|
<0,001
|
|
Answer
|
Question 11
(pre-administration)
|
Question IX
(post-administration)
|
|
0 (never/ rarely)
|
0
|
4
|
|
1 (sometimes)
|
14
|
40
|
|
2 (often)
|
22
|
6
|
|
3 (every day)
|
12
|
0
|
|
4 (many times a day)
|
2
|
0
|
|
|
Chi-square
|
|
|
Phi
|
Yates
|
Pearson
|
|
+0,61
|
34,52
|
36,95
|
|
P
|
<0,001
|
<0,001
|
|
Answer
|
Question 12
(pre-administration)
|
Question XII
(post-administration)
|
|
0 (yes, accept)
|
50
|
50
|
|
1(no, do not accept)
|
0
|
0
|
|
Answer
|
Domanda X (post-administration)
|
|
0 (no)
|
40
|
|
1 (yes)
|
10
|
|
Answer
|
Question XI (post-administration)
|
|
0 (epigastric pain)
|
2
|
|
1 (heartburn)
|
0
|
|
2 (headache)
|
2
|
|
3 (meteorism)
|
7
|
|
4 (halitosis)
|
2
|
|
5 (burping)
|
0
|
|
6 (urinary retention)
|
0
|
|
7 (night awakenings)
|
0
|
|
8 (other)
|
myalgia
|
As previously stated, Irritable Bowel Syndrome (IBS) is a gastrointestinal dysfunction caused by multiple “bio-psycho-social” factors which determine symptoms which may even be contrasting (constipation and diarrhea, soft feces alternating with hard feces) and which are often irritating, embarrassing, unpredictable, uncontrollable and painful.
Non biological markers of reference exist, therefore the diagnosis is made by exclusion of pathologies and by consulting the established symptomatic criteria.
This study recruited 50 patients (15 males and 35 females) who fell into the symptomology critera of IBS.
Through an initial questionnaire and one in the end, we wanted to verify if, after treatment with Kluyveromyces B0399 for three months, the symptomology changed.
The first question (pre-administration, question 1) determined the admission to the study. In the test-pre-administration (question 1) 50 patients gave an answer greater than 2, revealing to be affected by IBS; 29 of them (58%) gave 5 as an answer, which corresponds to a pain for more than one day a week. At the end of treatment, 37 patients (74%) had a score of less than or equal to 2, which would exclude them from the diagnostic criteria for IBS. The remaining 13 patients (26%) still had discomfort or pain for 2/3 days a month (8 patients) or for 1 day a week (5 patients).
These results are important, as shown by the statistical analysis which compares the answers that indicate a healthy person (0-1-2) and those which indicate the presence of IBS 3-4-5-6), revealing a definite, significant difference between the answers given before the treatment and those given after ( Chi-square Yates = 55,6 with p < 0,001; Chi-square Pearson = 58,73 with p < 0,001). The answers to the first question underline how treatment with Kluyveromyces marxianus fragilis B0399 drastically reduces the symptomology with the exclusion of most of the patients from the diagnostic criteria for IBS.
The second question (Is this discomfort present just during menstruation?)
(pre-administration question 2 and post-administration question II) remained unchanged. The answers of all the women revealed an independence of menstruation from the symptomology of IBS both before and after treatment.
The third question (Have you been suffering from this discomfort or pain for 6 months or more?)
(pre-administration question 3) showed that all the patients suffered from abdominal discomfort or pain for more than 6 months (chronic symptomology) and was not asked again in the final control because it’s a normal course for the symptomology of IBS.
The fourth question (How often does this discomfort subside or disappear after defecation?)
(pre-administration question 4 and post-administration question III) regarded the association between pain or discomfort and defecation. The statistical analysis showed that there is a significant difference (with p< 0,05) between the answers in the questionnaire pre-administration and the one post-administration ( Chi-square Yates = 4,58 with p = 0,032347; Chi-square Pearson = 5,54 with p = 0,018587).
Initially, 28 patients (56%) reported an improvement after defecation. At the end, 20 patients (40%) reported an improvement in pain after defecation. This is due to the improvement of the overall symptomology and therefore less evidence of an association between pain – defecation.
The fifth and sixth questions (analysis of the frequency of defecation in relation to abdominal discomfort or pain.)
(pre-administration questions 5 and 6) compared the relation of pain with a change of intestinal cavity (more frequent or less frequent defecation). 36 patients (72%) reported having diarrhea and 23 (46%) constipation. Apparently, some patients have a mixed form of IBS: both diarrhea and constipation (9 patients=18%).
In the questionnaire post-administration (question IV), all the patients reported an improvement with 64% (32 patients) reporting a normalized intestinal cavity.
The seventh question (frequency of soft feces)
(pre-administration question 7 and post-administration question V) analyzed the relationship between abdominal pain or discomfort and soft feces: In the first questionnaire, 14 patients reported having soft feces all the time or most of the time, while the other 22 had soft feces often or sometimes. After the treatment with Kluyveromyces, 18 patients (36%) didn’t have soft feces and 12 patients had them just sometimes. The statistical analysis of the data describes how the answers of the patients vary in the pre-administration questionnaire and the one post-administration, with a change from the answers that indicate an elevated frequency of soft feces (2-3-4) to ones that describe sporadic episodes (0-1) ( Chi-square Yates = 9,72 con p = 0,001823; Chi-square Pearson = 11,18 con p = 0,000827)
The eighth question (pre-administration question 8 and post-administration question VI) looked into the relationship between pain and hard feces. This symptom was present in 23 patients (46%), 13 of which had the symptom just sometimes and 8 others often or always.
After the treament (question V) the hard feces disappeared in 10 patients and were present in 8 patients just sometimes. Even though in this case the statistical analysis doesn’t show a significant difference between the pre-administration answers and those post-administration, the graph representing the data shows a rather evident change in frequency, with a diminishing presence of hard feces.
The ninth question (frequency of hard and lumpy feces)
(pre-administration question 9 and post-administration question VII) looked into the correlation between pain and hard and lumpy feces. In 9 patients (18%) the feces were often hard and in 8 they were hard most of the time or always. After the treatment (question VII), 12 patients reported not ever having hard feces and 8 only sometimes. The answers of the pre-administration test and post-administration test are significantly different, with a change in the answers that indicate an elevated frequency of soft feces (2-3-4) to ones that describe sporadic episodes (0-1) ( Chi-square Yates = 10,9 con p = 0,000962; Chi-square Pearson = 12,9 con p = 0,000329), indicating that the treatment with Kluyveromyces B0399 diminishes the frequency of hard and lumpy feces.
In the tenth question (frequency of loose, soft or watery feces)
(pre-administration question 10 and post-administration question VIII) the relationship between pain and loose and watery feces was investigated. 12 patients reported having this symptom sometimes while 22 patients reported having it most of the time or often. After treatment with Kluyveromyces marxianus fragilis B0399, 26 patients did not have watery feces any more and only 10 patients had them just sometimes (question VIII). The verification of the statistical analysis confirms a considerably significant change in the answers from those that indicate an elevated frequency (2-3-4) of soft feces to those that describe sporadic episodes (0-1) (Chi-square Yates = 21,64 with p < 0,001; Chi-square Pearson = 23,97 with p < 0,001), demonstrating that the appearance of loose or watery feces was diminished.
The eleventh question (How much does abdominal pain or discomfort influence social life?)
(pre-administration question 11 and post-administration question IX ) looked into the relationship between IBS and quality of life. The first questionnaire showed that 34 patients (68%) were conditioned often or every day by IBS and 14 patients were conditioned sometimes. After the treatment (question IX) 40 patients (80%) reported being conditioned in their social life only sometimes. The verification of the statistical analysis confirms a considerably significant change in the answers, from those that indicate a considerable discomfort (2-3-4) to ones that describe a light or discomfort or none at all (0-1) (Chi-square Yates = 34,52 with p < 0,001; Chi-square Pearson = 36,95 with p < 0,001).
Beyond all the symptomatic improvement demonstrated by the above questions, this information leads to considering the importance that treatment with Kluyveromyces marxianus fragilis B0399 can have in mutating not only physical factors but psychological ones as well in patients with IBS.
In the post-administration questionnaire a question was added that inquires about eventual side effects (X). Ten patients (20%) answered in the affirmative, specifying the particular symptom in the following question (XI): 7 patients reported having halitosis, 2 headache, meteorism, burping, heartburn, 1 epigastric pain and myalgia. No one had allergies, intolerance or other symptoms.
The last question (corresponding to XII) had the purpose of informing the patients of the collection and use of data (though anonymous) and getting their permission (100% of the patients accepted).
Acknowledging the fact that IBS has multiple causes (physiological, psychological, environmental and behavioral) and therefore there is no single effective therapy but multiple therapies which are aimed at the most important subjective causes, this study, based on the symptomological data gathered before and after the treatment, demonstrates that the prolonged administration of Kluyveromyces marxianus fragilis B0399 significantly improves the clinical condition and above all improves the quality of life of the patients.
A correct diet, together with physical exercise and stress control, further improve the situation, giving the doctor the job of informing the patient, educating him and giving him confidence and then prescribing the most effective drug.
BIBLIOGRAFHY:
