Alvah C. BITTNER, PhD, CPE1,2    Dra. Rocio CAICEDO, B.3

Dr. Telmo DE la TORRE, C.4, 5       Titut N. YOKELSON, PhD6

Clinical research was conducted to evaluate the efficacy of Prescript-Assist™ (P-A), an advanced probiotic-prebiotic formulation for treatment of human bacterial-diarrhea in two communities in Ecuador. P-A has previously demonstrated efficacy against: Canine bacterial-diarrhea in short-term studies; and Irritable-Bowel-Syndrome and related gastric diseases in both short-term and longer-term human studies (e.g., 1-year follow-up). In this study, P-A treatment was shown to result in significantly shorter diarrhea-durations than standard-antibiotics used in treating bacterial diarrhea [χ2(4) = 41.1, P<2.6•10-7]. Prescript-Assist™ and other advanced probiotic-prebiotic combinations have considerable promise as emerging treatments for a wide spectrum of both acute and chronic GI diseases.

This report describes our recent clinical experience using an advanced probiotic-prebiotic formulation for treatment of human bacterial-diarrhea in Ecuador [Prescript-Assist™ (P-A): Safer Medical, Inc. (SMI), Ft. Benton, MT].1 The P-A complex previously was found particularly effective for the treatment of a wide variety of human GI disorders and bacterial diarrhea.[1,2,3,4] The particular breadth of efficacy – relative to earlier probiotic treatments – is attributed to the actions of a unique complex of >29 probiotic soil-based-organisms (SBOs) and leonardite, a prebiotic mix of humic substances that differentially enhance SBO proliferation.[1] Among conditions initially reported as clinically responding to the P-A probiotic-prebiotic complex were both: "Occasional diarrhea" (e.g., travelers); and "Chronic diarrhea" such as associated with Irritable Bowel Syndrome (IBS) and Colitis.[4] Early clinical reports[4] prompted a formal randomized, placebo-controlled, double-blind clinical study with regard to IBS.[3] This placebo-controlled study identified 3 relatively-independent subsyndromic factors of IBS: General ill-feelings/nausea, Indigestion/flatulence, and Colitis. More importantly, the combined probiotic-prebiotic treatment (P-A, 550mg BID) was associated with broad reductions in each of the subsyndromic factors (Ps <0.04) – and their associated symptoms – in a two-week double-blind study in patients with IBS.[1]

The reduction in "Colitis" – associated in the literature with a C. difficile infection – arguably supported the earlier indications of P-As potential as a treatment for occasional diarrhea.[5] This, together with the earlier clinical observations, encouraged both: Follow-up of P-A’s efficacy in the IBS study population[2] and Evaluation of P-A’s specific efficacy for bacterial-diarrhea in animal patients.[3] In the remainder of this section, the results of these efforts are introduced both as part of the "background" for the current effort and to set the stage for the "purpose."

Bacterial diarrhea is one of the most common maladies faced by medical care-givers working in the third-world. In humans, and their canine companions, a number of bacteria have been commonly associated with diarrhea, including: Salmonella, C. perfringens, and C. difficile. However, these and a variety of other organisms are not uncommonly a part of "healthy intestinal microflora." Hence, stool studies and related diagnostic approaches may be neither timely nor useful when faced with an accelerating outbreak of diarrhea in a closed population. Medical emphases consequently include: immediate treatment to limit the duration of diarrhea; and other actions to limit its possible transmission (particularly as untreated-diarrhea is associated with up to 20% deaths in the young).

The seriousness of bacterial diarrhea and earlier reports of efficacy against "occasional diarrhea" prompted an exploratory study with regard to canine bacterial-diarrhea.[3] The P-A complex [marketed for pets as PetFlora™ Vitality Sciences Inc, Oakland Park, FL] was administered (250mg BID) to 10 dogs suffering from bacterial diarrhea during a kennel outbreak. In response to this outbreak, kennel areas and animals were first thoroughly cleansed and all animals were immediately administered PetFlora™ (as a prophylactic for those not showing symptoms). It is noteworthy that both food and water were made available to all – past experience suggesting that diarrhea could be readily controlled without restrictions. No signs of diarrhea were seen after 12 hours observation – this was certainly less-than the median of the 24-to-48 hours typical with a traditional antibiotic treatment (Neomycin generic "Biosol", 1.1cc/10kg, with food and possibility water restrictions). This [PetFlora™ vs. antibiotic] difference in diarrhea-duration of was very-highly significant (P<0.002, 2-tail, Binomial Sign-Test).[3, 6]

Supporting these "antibiotic-like actions" were observations of complete remission of IBS symptoms at one-year follow-up of patients maintained open-label for a 2 to 4 weeks period immediately following a double-blind study (4-6 weeks total).[2] Continuing remissions were observed in 62.5% in a group – at the 1-year follow-up – without further support (5/8), and in 100% of those (6/6) choosing maintenance P-A treatment [ultimately at reduced dosing, e.g., one cap every 1-3 days]. The final group of 10 patients – to generally good effect – chose intermittent use of P-A only when signs of IBS symptoms began to reappear. 2

The body of past studies altogether encouraged our initiation of clinical studies of bacterial-diarrhea in populations at risk – the present study is represents a first step in this direction.

The body of this section first delineates methodological aspects of the study: Participants and Test-Instrument. This is followed by a joint consideration of analytic methods and their principal findings. This sets the stage for the final section – Discussion and Conclusions.

The study population was comprised of 21 patients (12♂ and 9♀) – presenting with persistent bacterial-diarrhea symptoms – almost equally drawn from two agrarian mountain communities located in mountainous regions about Quito, EC. Miraflores, served by Dra. Caicedo during the research study, is located 2.5 hours South of Quito; whereas, Malchinqui, about 1.25 hours North toward Cayambe, was served by Dr. De la Torre. Both communities – not surprising given continuing waves of diarrhea – were found to have bacterially contaminated public or well-water sources (e.g., Fecal Coliforms).3 Patients – reflecting the largely indios-mestizo ethnicities of their communities – ranged in age between 2 to 65 years.4

Recorded for each participant were: Name, Age, Community, Sex (♂ or ♀), and Physician Observations/Comments. Six primary symptoms also were assessed on a daily basis: Diarrhea, Gases, Abdominal pain, Constipation, Vomiting, and Fever. For research purposes, these symptoms were scored according to the number of days they were observed – starting with "1" at day of clinical diagnosis. Breaks – in any symptom – were conservatively scored as if present, if the symptom subsequently returned during the observational period. Using this approach each of the six symptoms was scored for total days duration.

Analyses were conducted in three phases. During the Phase I, a Principal Factor Analysis (PFA) was conducted of the six clinical symptoms in a patients receiving P-A Treatment.[7,1] This provided for an exploratory analysis of dimensional "syndromic-factor" structure of the symptoms – akin to that reported in our earlier study of IBS.[1] In keeping with this earlier study, Analyses-of-Variance (ANOVAs) were planned for applications to any emergent sub-syndromic factors (with a focus on the main effects of Place-Community, Age, and Sex).[8,1,2] During Phase II, ANOVAs were conducted to evaluate the effects of Place (community), Sex, Age and selected interactions on the log-transformed duration-of-diarrhea: Ln(D). Phase III follows this with final analyses comparing the log-transformed durations-of-diarrhea with P-A vs. community-specific experiences with standard antibiotic treatments.

3 Contaminated food(s), perhaps not unrelated to water issues, were occasionally mentioned as the source of illness/diarrhea in some patient reports.

4 SMI’s IRB had approved the study protocol, which provided for incrementally extending treatment of outside the initial 5-65 year age-limits – if/as clinical experience supported efficacy and safety of P-A. No patients <2 years were treated for diarrhea during the study – this was reportedly was due to: the relative absence of the disease in infants (purportedly due to their being breast-fed till 2 years-of-age).

PFA revealed two factors that accounted for 68.8% of the total symptom variation. Table 1 presents the rotated factor matrix where it may be seen that the first factor "F1-Diarrhea-Vomiting" is essentially defined by loadings on Diarrhea(0.83) and Vomiting (0.88) with lesser though significant contributions by Fever (0.62) and Nausea (0.52). The second factor "Gas/Abdominal-Pain" is defined by almost identical loadings on these symptoms: Gas (0.92), Abdominal-Pain (0.92) with no significant contributions by other variables.

A secondary goal – of the present study – was to explore the multivariate nature of the presenting diarrhea symptoms, particularly toward selecting measure(s) of disease severity and efficacy. As seen in the Phase I PFA results, some progress was made toward this end. Our findings essentially identified a unitary factor with significant loadings by all but Gas and Abdominal-Pain (which defined a second factor without much in the way of significant relationships with the experiment variables of interest). The dominant first factor "Diarrhea-Vomiting" was: (a) Found related to variables of interest; and (b) Broadly defined by loadings on Diarrhea (0.83) and Vomiting (0.88) with lesser though significant contributions by Fever (0.62) and Nausea (0.52). The prominent Diarrhea (0.83) loading – as well as its primary clinical significance – supported a focus on this variable during the analyses of Phase II.