The Ruggiero-Klinghardt (RK) Protocol for the Diagnosis and Treatment of Chronic Conditions with Particular Focus on Lyme Disease


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The Ruggiero-Klinghardt (RK) Protocol for the Diagnosis and Treatment of Chronic Conditions with Particular Focus on Lyme Disease

Dietrich Klinghardt and Marco Ruggiero

Sophia Health Institute and Klinghardt Academy, Woodinville, WA., USA


Article history

Received: 20-02-2017 Revised: 28-02-2017 Accepted: 08-03-2017

Corresponding Author:
Marco Ruggiero
Sophia Health Institute and Klinghardt Academy, Woodinville, WA., USA

Abstract: Here we describe the Ruggiero-Klinghardt (RK) Protocol that is based on integration of Autonomic Response Testing (ART) with diagnostic ultrasonography and on application of therapeutic ultrasounds; the latter are used as a provocation tool and as an instrument to optimize drug uptake and utilization in specific areas of the body. This protocol consists of a precise sequence of diagnostic and therapeutic procedures with the ultimate goal of improving sensitivity and specificity of diagnosis at the same time evaluating and optimizing efficacy of treatments in chronic conditions including, but not limited to, persistent Lyme disease. The RK Protocol represents a paradigm shift in diagnostics and therapeutics: Thus, compartmentalized microbes, transformed cells, toxins and metabolites could be detected using a safe and non-invasive method. In addition, the RK Protocol allows optimization of efficacy of drugs and other therapeutic interventions. Although the RK Protocol was initially developed for persistent Lyme disease, it shows significant potential in conditions ranging from cancer to neurodegenerative diseases and autism. In oncology, the RK Protocol may serve to facilitate early diagnosis and to increase sensitivity of cancer cells to the killing effects of a variety of remedies ranging from conventional radio- and chemotherapy to more recent forms of immunotherapy. Thus, the 1st goal of the RK Protocol is diagnostic: That is, to make pathogens, toxins, transformed cells and cells infected by viruses that are inaccessible to conventional diagnostic and therapeutic tools, “visible” to the therapist who can detect them with laboratory methods and deal with them with appropriate interventions; and also to make them “visible” to the immune system that can fight them in a physiological manner. The 2nd goal is to optimize drug uptake and utilization in the organs and tissues studied and targeted with these procedures.

Keywords: Lyme, Ultrasound, Autonomic Response Testing, Immune System, Imaging, Brain



The infectious disease known as Lyme borreliosis, or Lyme disease, is the most common infection due to tick bites and sometimes also to other stinging insects in the Northern Hemisphere. Although estimates vary and it is likely that the number of cases worldwide is much higher, there is general consensus that the disease affects hundreds of thousands of individuals a year in North America, Europe and northern Asia with incidence of the infection on the rise (Shapiro, 2014;

Gingrich et al., 2001). The designation “Lyme disease” derives from the small New England villages of Lyme and of Old Lyme where the arthritic manifestations of the disease were first described in 1975 as “a rather random clustering of several cases of juvenile chronic arthritis” (Burmester, 1993).

The etiologic infectious agent, Borrelia burgdorferiwas identified by Wilhelm “Willy” Burgdorfer it in 1981 and, since the original observation limited to arthritis, the clinical spectrum of the infection now includes a variety of clinical conditions such as erythema migrans,

American Journal of Immunology


© 2017 Dietrich Klinghardt and Marco Ruggiero. This open access article is distributed under a Creative Commons Attribution (CC-BY) 3.0 license.

Dietrich Klinghardt and Marco Ruggiero / American Journal of Immunology 2017, 13 (2): 114.126

DOI: 10.3844/ajisp.2017.114.126

acrodermatitis chronica atrophicans, lymphadenosis benigna cutis, arthritis, myocarditis, neuroborreliosis (meningo-encephalitis, meningo-radiculitis, meningitis), myositis and various ocular and skin disorders (Burgdorfer, 1991; Chomel, 2015). Interestingly, as knowledge of the disease deepened, the increase in the number of clinical manifestations of the infection was paralleled by the increase in the number of spirochete bacteria belonging to the genus Borrelia associated with the disease; Borrelia burgdorferi sensu stricto is found in the Americas whereas Borrelia afzelii and Borrelia garinii, in addition to Borrelia burgdorferi, are observed in Europe and Asia (Chomel, 2015). In a sort of an exponential increase in complexity, it was later discovered that the ticks responsible for transmitting theBorreliae that are Ixodes scapularis and Ixodes pacificus, also have the potential to transmit an increasingly expanding list of other pathogenic microbes that include bacteria, viruses and parasites such asAnaplasma phagocytophilum, Babesia microti, deer tick (Powassan) virus, Borrelia miyamotoi and the Ehrlichia muris-like organism (Caulfield and Pritt, 2015). Obviously, the presence of coinfections renders the treatment and the diagnosis rather difficult and contributes to patient morbidity and mortality as well as to the appearances of the so called “post-treatment Lyme disease” also known as “chronic Lyme” or, as we prefer to denominate it in this study, “persistent Lyme disease”. Thus, persistence of Borrelia burgordferi in tissues after efficient antibiotic treatment has been demonstrated in a variety of experimental models that include primates, but there is currently no affordable, non-invasive, method to detect specific persistent microbes (or their metabolites) in vital organs such as the brain, spinal chord or heart (Straubinger et al., 1997; 2000; Hodzic et al., 2008; Yrjänäinen et al., 2010; Embers et al., 2012).

Persistent Lyme is a diagnosis that is given to patients with prolonged, unexplained symptoms, that may be severe, occurring after efficient antibiotic treatment. Such symptoms are most often non-specific and include neurological symptoms such as chronic fatigue, difficulty to sleep, inability to focus, irritability, depression, headache and dizziness; gastrointestinal symptoms such as abdominal pain, nausea and diarrhea; symptoms suggestive for local or systemic inflammation such as pharyngodynia, enlarged lymphnodes, muscle rigidity, myalgia and arthralgia (Feder et al., 2007). The non-specificity of these symptoms and the absence of reliable laboratory tests has led to questioning the very existence of a persistent Lyme disease since it is claimed that there is no strict evidence that the chronic post- treatment symptoms are attributable to ongoing infection with Borrelia burgdorferi or with any other identified organism (Feder et al., 2007; Halperin, 2015). However, despite the controversies surrounding the existence, the

definition or the nature of persistent Lyme, it is indisputable that this condition leads to significant sequelae that dramatically decrease the quality of life of affected patients. Thus, a recent study from the John Hopkins School of Medicine describes the physical and social limitations associated with the condition that lead to fundamental changes in the way of living. In this study, the Authors evidence the disease-specific factors that contribute to symptom and illness invisibility and the pervasive medical uncertainty regarding persistent Lyme that promotes an increased sense of personal responsibility for care. The Authors conclude that “similar to other contested or medically unexplained syndromes, our findings suggest that the social sequelae of post-treatment Lyme disease/chronic Lyme can be equally protracted as the physical effects of this illness” (Rebman et al., 2015).

The main reason for the contested status of persistent Lyme as a fully recognized disease mainly resides in the objective difficulty to assess the persistence of Borrelia, or other pathogens responsible for coinfections, after efficient antibiotic treatment. However, persistence ofBorrelia burgordferi in tissues after antibiotic treatment has been demonstrated in a variety of experimental models, which include primates, that were infected withBorrelia burgdorferi and then received aggressive and successful antibiotic treatment for 4-6 months (Straubinger et al., 1997; 2000; Hodzic et al., 2008; Yrjänäinen et al., 2010; Embers et al., 2012).

The latter study demonstrates that the pathogen is able to resist antibiotic treatment in primates, even though the Borrelia in itself is not known to possess resistance mechanisms and it is susceptible to common antibiotics such as doxycycline and ceftriaxone in vitro. Therefore, mechanisms other than classic antibiotic resistance have been hypothesized to explain the persistence of the bacteria and these include the formation of drug-tolerant persister cells (Sharma et al., 2015), cell wall deficient forms and biofilm residing forms, as well as the generation of cystic forms of Borrelia with low metabolic activity that enables the spirochete to survive in a hostile environment until conditions are favorable to proliferate again (Murgia and Cinco, 2004).

The controversies and the difficulties associated with the diagnosis and treatment of persistent Lyme could be resolved by the existence of reliable and reproducible tests that were able to ascribe the symptoms to chronic infections rather than to a wide number of other conditions that may present themselves with similar or identical symptoms (Eshoo et al., 2013; Curcio et al., 2016).

At Sophia Health Institute, we use a manual biofeedback technique, denominated Autonomic Response Testing (ART) that has the goal of assessing



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the presence and/or the persistence of spirochete and other infectious agents that may be associated with persistent Lyme disease. ART represents an evolution and an expansion of the technique that was originally proposed by Omura (1981) and subsequently validated in a number of studies including two randomized-order blinded studies registered as a clinical trial (Jacobs et al., 1984; Jensen et al., 2016). In a paper published in 2016, ART developed at the Sophia Health Institute was utilized by independent researchers in the diagnosis and treatment of painful scars. In this study, the Authors state that “In our experience, ART produces useful and consistent information most of the time” (Chung and LaRiccia, 2016). The validity of ART was further confirmed in another independent investigation focused on breast cancer (Chung and LaRiccia, 2017).

In addition to ART, at Sophia Health Institute, we investigate the presence of pathogens in urine samples of patients using Polymerase Chain Reaction (PCR) in order to identify genes specific for microbes known to be associated with persistent Lyme. However, we noticed quite often that, while ART suggested the presence of pathogens, such a presence was not confirmed by the PCR-based DNA test performed on urine samples. Other researchers have noticed the low sensitivity of the urine DNA test when it is performed without the provocation method that will be described in the following sections of this study. However, serendipitous observation at Sophia Health Institute suggested that the results of the urine DNA test matched more closely the results of ART when urine samples were collected after the patient had received a deep tissue massage with focus on the symptomatic body regions; massage that had been performed to relieve tension or for other reasons. Based on these observations and in search for a reliable method to improve the sensitivity of the urine DNA test, we decided to apply pulsed therapeutic ultrasounds focused on critical areas to mimic or improve the effects of deep tissue massage. This approach is based on the widely recognized feature of ultrasounds to transmit waves of compression and relaxation in biological tissues with resulting changes in organ, cellular and molecular structures that can be exploited in the context of therapy (Leinenga et al., 2016). Thus, we have recently demonstrated that non-thermal ultrasounds induce quasi-instantaneous changes in human neurons and murine microglial cells in vitro with results that are consistent with the observed effects of ultrasounds on mental states (Cosentino et al., 2015; Bocchi et al., 2015; Hameroff et al., 2013).

Here we describe an original protocol denominated Ruggiero-Klinghardt (RK) Protocol that is based on ART and on a precise sequence of diagnostic and therapeutic ultrasounds for the accurate and reliable

diagnosis and treatment of persistent Lyme disease and other chronic conditions.

Materials and Methods

Ultrasound Systems

For diagnostic ultrasonography, we used a portable MicroMaxx ultrasound system manufactured by Sonosite, Bothell WA, USA, with color-doppler application and with a linear (L38e) and a convex (C60e) transducer. This system is approved for many applications including cephalic (brain) imaging and has the same features of the system that one of us (M.R.) had previously used to characterize the lesions in the brains of autistic children (Bradstreet et al., 2014); in this latter study, the safety of the procedure is thoroughly described. In order to avoid any bias, the images were not exported and pictures of the screen of the ultrasound system were taken with the camera of a common smartphone. This procedure was chosen in order to avoid the possibility of editing the images, that are presented here exactly as they were produced; this procedure, however, inevitably leads to poor quality images since these were taken at the bedside without any consideration for professional photo-shooting. An example can be observed in Fig. 4, where portions of the screen of the ultrasound system can be observed. The pictures of the images were then copied in a Power Point file in order to add captions, arrows or circles that are useful to show the anatomical structures that were studied and their alterations. The application of therapeutic ultrasounds, that is an essential part of the RK Protocol, was performed using an Intelect TranSport Ultrasound Therapy device (Chattanooga Medical Supplies Inc., Chattanooga TN, USA). This system enables pulsed and continuous operation (10, 20, 50 and 100%) on 1 and 3.3 MHz frequencies using a 5 cm2 sound head applicator.

Urine DNA Test

Urine samples were collected before and after application of therapeutic ultrasounds at Sophia Health Institute according to the protocol summarized in Table 1 and following the recommendations of the company performing the DNA test. The test was performed by DNA Connexions, Colorado Springs CO, USA. The Lyme panel tests for 4 different genes that are found inBorrelia burgdorferi and 8 common Lyme disease co- infectors including Babesia microti, Babesia divergens,Babesia duncani, Bartonella bacilliformis, Bartonella henselae, Bartonella quintanta, Borrelia miyamotoi,Borrelia recurrentis, Ehrlichia chaffensis andAnaplasma phagocytophilum. According to the company and consistent with common knowledge, a positive PCR- based Lyme test indicates the presence of DNA fromBorrelia burgdorferi and/or other co-infectors.



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Table 1. Step 1. Step 2. Step 3. Step 4.

Step 5.

The Ruggiero-Klinghardt (RK) Protocol steps of diagnostic and therapeutic procedures
First comprehensive Autonomic Response Testing (ART).
Total-body diagnostic ultrasonography that has the role of further refining the diagnostic hypotheses put forward by ART. Application of therapeutic ultrasounds with particular focus on the areas identified as “abnormal” with the previous steps. Second ART performed after application of therapeutic ultrasounds: Comparison of the results with those obtained with the first ART of Step 1.
Collection of midstream urine in a sterile container that is then shipped to the laboratory. Alternatively, the presence of microbes, toxins, circulating cancer cells or other pathogenic noxae is investigated in other biological matrixes such as stools, blood or serum or breath that are appropriately collected.
Pharmacological treatment with remedies specific for the identified pathogens.
Daily application of therapeutic ultrasounds targeted to the areas identified with the previous steps.


Step 6.
Step 7.
Follow-up at 3-4 month intervals to evaluate the effectiveness of treatment and to assess the treatment end-point.


A negative result, however, does not prove that a patient is not infected with a tick borne infection, rather it indicates the absence of detectable DNA pertaining to microbes associated with Lyme and/or other tick borne co-infections in that particular urine sample. Stage of infection, timing of courses of antibiotics and persistence of latent microbes in reservoirs or sanctuaries, are only some of the factors that may affect the detectability of the DNA of spirochetes in urine samples.

Autonomic Response Testing (ART)

ART, invented and developed by one of us (D.K.), represents an evolution and an expansion of the test originally described by Omura (1981).

At variance with the test proposed by Omura, ART takes into account the entirety of the autonomic response and not only the strength or the resistance of muscles. This is particularly important in the study of parasympathetic activity and in the evaluation of the balance between sympathetic and parasympathetic activities. A core principle of the test relies on the “Resonance Phenomenon Between Identical Substances”. Thus, a culture of a particular pathogen is used to non-invasively detect the presence of this very pathogen in a particular body region or organ. A description of ART can be found in a recently published peer-reviewed study (Chung and LaRiccia, 2016). In the present study, as per the RK Protocol, ART was performed two times, i.e., before and after application of therapeutic ultrasounds as summarized in Table 1; the results of the two tests were then compared and recorded. Since interpretation of ART can provide information on the potential responsiveness of the patient to different therapeutic approaches and since the involvement of the immune system in persistent Lyme is widely acknowledged, we used an emulsion of chondroitin sulfate, vitamin D3 and oleic acid endowed with immune modulating properties as positive control (dr. reinwald healthcare, Schwarzenbruck, Germany). The characteristics of this emulsion and its potential use in integrative immunotherapy have been recently described (Schwalb et al., 2016).



Description of the RK Protocol Step 1

After having collected a careful anamnesis and critically reviewed clinical records, laboratory exams and radiological images, the patient is invited to empty her/his bladder before performing the first step of the RK Protocol that consists in ART which is performed by the therapist with the help of an assistant; the sequence of steps of the RK Protocol is summarized in Table 1. ART provides the initial information that is useful to restrict the spectrum of diagnostic hypotheses and to identify the organs or the areas of the body that need further investigation.

Step 2

Then, as the second step of the protocol, a diagnostic total body ultrasonography is performed with particular focus on those areas that have tested positive with ART.

The following examples elucidate the synergy between ART and ultrasonography in diagnostics. Figure 1 shows the ultrasonographic appearance of the submandibular salivary gland of a subject who had tested positive with ART in that area. The inhomogeneous appearance of the gland with irregular hypoechoic areas is indicative of a diffuse inflammatory process whose origin (viral or autoimmune) requires further investigation. Figure 2, shows an enlarged and possibly inflamed, deep cervical node in a subject with symptoms of neuroborreliosis who had tested positive with ART. The blood vessels in the hilum of the node are clearly visible at the echo-color-doppler and their appearance is consistent with a condition of hyper-afflux. It is worth noticing that inflammation of the deep cervical nodes may be associated with impaired lymphatic drainage from the brain lymphatic system (also known as “glymphatic system”) with consequent stagnation of lymph in the brain and accumulation of metabolites and neurotoxins in addition to potential disruption of

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the brain microbiota. These events may be associated with, if not responsible for, some of the symptoms of neuroborreliosis, autism and other neurological diseases (Bradstreet et al., 2014; Ruggiero, 2016). Figure 3, shows the appearance of the vagus nerve in a subject who had tested positive with ART for parasympathetic imbalance. In this transversal projection, the vagus nerve appears as a small triangular structure located posteriorly inside the carotid sheath between the common carotid artery and the internal jugular vein; it shows an internal honeycomb structure. In this subject, the epineurium appears as a thickened hyperechoic ring surrounding the nerve. Figure 4, shows the appearance of the thyroid in longitudinal projection in a subject who had tested positive with ART for parathyroid involvement. The arrow indicates a roundish area that protrudes from the posterior margin of the thyroid and could be interpreted as an enlarged parathyroid gland. Please notice that the solid arrow was inserted during the ultrasound examination whereas the dotted circle was inserted during the preparation of the figure, for the sake of clarity, using a Power Point program.

Step 3

After having performed the diagnostic total body ultrasonography, the third step of the RK Protocol is then performed; therapeutic ultrasounds are directed toward those organs or areas of the body where clinical suspicion of inflammation, proliferative, degenerative or infectious diseases is present. The rationale for this procedure is to exploit the mechanical effects of ultrasounds in human tissues with the goal of mobilizing pathogens, toxins, transformed cells, cells infected by viruses, or cells of

the immune system carrying pathogens, toxicants or antigens of transformed cells, so that they may be identified by the second ART and by specific tests such as the urine DNA test. In other words, the goal of application of therapeutic ultrasounds is to force the exit from reservoirs or sanctuaries where pathogens or transformed cells are invisible to the immune system and to the therapist performing ART or using laboratory methods and are protected from therapeutic intervention (Cory et al., 2013). Choice of the pulsed sequence and frequency depends on the location of the organ to be treated.

For example, when treating the spleen, a major organ of the immune system, we choose a pulsed sequence indicated as 50% and a frequency of 1 MHz. First the spleen is identified by diagnostic ultrasonography (Fig. 5) and then it is treated for 3 min, slowly moving the sound head applicator so to send the ultrasound waves to most areas of the organ. The effectiveness of application of therapeutic ultrasounds is then assessed by studying the blood flow in the spleen after the treatment, using for this purpose the diagnostic echo-color-doppler technique. As shown in Fig. 6, immediately after application of therapeutic ultrasounds, a significant increase in blood flow can be observed at the diagnostic ultrasonography. We have previously demonstrated that such an increase corresponds to the activation of cells of the immune system inside the spleen, with particular reference to macrophages (Ruggiero et al., 2014). Thus, the mechanical waves of compression and relaxation of therapeutic ultrasounds “squeeze” the organ, presumably at the level of the microscopic anatomy of spleen as well as at the level of cellular and molecular structures such as the proteins of the cytoskeleton (Hameroff et al., 2013).


Fig. 1. Ultrasonographic appearance of the submandibular salivary gland118

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Fig. 2. Ultrasonographic appearance of a deep cervical node


Fig. 3. Ultrasonographic appearance of the vagus nerve


Fig. 4. Ultrasonographic appearance of the thyroid119

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Fig. 5. Ultrasonographic appearance of the spleen before therapeutic ultrasound treatment

Fig. 6. Blood flow in the lower pole of the spleen before and after application of therapeutic ultrasounds


We hypothesize that mobilization of pathogens or toxins from sanctuaries or reservoirs is followed by their elimination through the urine and this phenomenon should optimize the sensitivity of the urine DNA test. In addition, we have observed that the sensitivity and the specificity of ART significantly increase after application of therapeutic ultrasounds, thus improving the intra- and inter-operator reproducibility of the technique. Furthermore, as we have observed using the immune modulating emulsion mentioned in the Materials and Methods, application of therapeutic ultrasounds to a specific organ significantly improves the sensitivity of the organ to the remedy. Such an effect did not come as a surprise since it is known that ultrasounds have the potential to increase the effectiveness of a number of drugs and remedies by favoring their cellular uptake and overcoming the barriers that prevent delivery of drugs to specific lesions (Bui et al., 2017).

We observed that the biological effects of application of therapeutic ultrasounds were particularly remarkable when the technique was applied to the brain, with particular reference to the temporal lobes. We and others

have previously demonstrated that ultrasounds could be directed toward specific areas of the brain using carefully selected acoustic windows such as the temporal squama (Ruggiero et al., 2013; Hameroff et al., 2013; Bradstreet et al., 2014).

More recently, we have demonstrated that ultrasounds elicit cellular and molecular responses in neurons and glial cells that are consistent with their effects on mental states and can be exploited in the context of therapy (Cosentino et al., 2015; Bocchi et al., 2015; Hameroff et al., 2013; Leinenga et al., 2016). Based on these evidence, we incorporated transcranial ultrasonography in the RK Protocol. Thus, after having performed the first ART of step 1, a diagnostic ultrasound scan of the brain is performed as shown in Fig. 7. This figure shows the squama of the temporalis bone as a homogeneous hyperechoic structure; the meninges are visualized as a series of alternating hyper- and hypoechoic layers with a regular structure; the subarachnoid space appears as an anechoic line due to the presence of extra-axial fluid; and the gray matter of the temporal lobe (in this case, the right temporal lobe)


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appears hypoechoic in comparison with the meninges showing a layered structure that corresponds to the cellular architecture of the cerebral cortex described by von Economo and Koskinas (1925). Study of the subarachnoid space may be used to evaluate the accumulation of extra-axial fluid according to the technique described in Bradstreet et al. (2014). However, at variance with our previous studies, at Sophia Health Institute, we do not limit our intervention to the evaluation of extra-axial fluid accumulation and how it relates to neurological symptoms, but we use that information to proceed with application of therapeutic ultrasounds to the brain and the deep cervical nodes (Fig. 2) with the intent of restoring the flow of cerebral lymph and possibly ameliorating neurologic symptoms when present (Bradstreet et al., 2015; Tarasoff-Conway et al., 2015).

In order to reach this goal, the deep cervical nodes that have been visualized by diagnostic ultrasonography are treated with therapeutic ultrasounds using a pulsed sequence indicated as 20% and a frequency of 3.3 MHz for 90 sec on each side of the neck. Only after having treated the nodes, the brain is treated with therapeutic ultrasounds that are administered through the temporal acoustic window using a pulsed sequence indicated as 10% and a frequency of 3.3 MHz for 90 sec on each side of the head. The rationale for this sequence of therapeutic procedures lays in the consideration that inflamed deep cervical nodes may pose an obstacle to the efflux of the lymph from the brain and, therefore, it is indicated to treat the deep cervical nodes first in order to reduce inflammation by exploiting the anti-inflammatory effects of pulsed ultrasounds (Jia et al., 2016). Subsequent treatment of the brain with therapeutic ultrasounds thus sends mechanical pressure waves consisting of alternate compression and relaxation that may favor the circulation of lymph in the brain

lymphatic system and the removal of catabolites and toxins that may have become stagnant in the presence of an obstacle to the circulation of lymph (Raper et al., 2016). It is worth noticing that, given the anatomical proximity of the two structures (Fig. 2 and 3) application of therapeutic ultrasounds to the deep cervical nodes may correspond to treatment of the vagus nerve as well. Thus, it is well assessed that vagus nerve stimulation provides a number of benefits ranging from treatment of affective disorders in psychiatry (Cimpianu et al., 2017) to improvement in recovery from traumatic brain injury (Neren et al., 2016). Not surprisingly, the second ART, performed after each therapeutic ultrasound treatment, showed changes consistent with the effects described above.

Step 4

The fourth step of the RK Protocol consists in the second ART that is performed after application of therapeutic ultrasounds and in the comparison of the results with those obtained with the first ART of Step 1.

Step 5

The fifth step of the RK Protocol consists in the collection of the urine samples that has to be performed after the application of the therapeutic ultrasounds; the patient collects the naturally occurring next midstream urine in the sterile container provided by the laboratory performing the urine DNA test and the sample is shipped overnight to the laboratory. Although in this study we describe the results obtained with the urine DNA test, the presence of microbes, toxins, circulating cancer cells or other pathogenic noxae can be investigated in other biological matrixes such as stools, blood or serum or breath that are appropriately collected and analyzed by specialized laboratories.


Fig. 7. Ultrasonographic appearance of the brain at the level of the right temporal lobe121

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Step 6

Successively, as the sixth step of the RK Protocol, the patient is treated with specific remedies that are targeted, for example, toward the pathogens identified with the previous steps. When antimicrobials are used, their choice is determined by applying common current understanding and fine tuning the selection with ART.

Step 7

The seventh step of the RK Protocol is performed in conjunction with the previous one and consists in the targeted application of therapeutic ultrasounds; such a procedure is performed once daily in the same manner described above in order to exploit the known therapeutic effects of pulsed ultrasounds that comprise anti-inflammatory effects, enhanced lymphatic drainage and optimization of drug uptake and utilization.

The RK Protocol is then repeated after three to four months in order to evaluate the effectiveness of treatment and to assess the treatment end-point.


In this study we describe for the very first time a protocol, the RK Protocol, that is based on the integration of ART, as a screening tool and ultrasounds; the latter are utilized as a diagnostic tool, as a provocation tool and as a means to optimize drug uptake and utilization in specific areas of the body. This protocol consists of a precise sequence of diagnostic and therapeutic procedures with the ultimate goal of improving sensitivity and specificity of diagnosis and evaluating and optimizing efficacy of treatments in chronic conditions including, but not limited to, persistent Lyme. The sequence that we have developed is summarized in Table 1 and comprises, as a first step, a comprehensive ART screening, followed, as a second step, by total-body diagnostic ultrasonography that has the role of further refining the diagnostic hypotheses put forward by ART. Diagnostic ultrasonography also has the role to precisely identify the anatomic localization of the organs or the structures toward which the pressure waves of the therapeutic ultrasounds will then be directed in the third step of the protocol.

Application of therapeutic ultrasounds, the third step, plays three roles in the RK Protocol:

  • It is used to exploit the well-known therapeutic effects of ultrasounds that have been observed in a variety of conditions associated with chronic inflammation or neurodegeneration (Jia et al., 2016; Leinenga et al., 2016)
  • It is used to increase the potential effectiveness of remedies, an effect that can be further verified by the second, post-ultrasound ART (step 4). This

phenomenon is due to the known efficacy of therapeutic ultrasounds to increase the uptake and utilization of remedies in cells and tissues (Bui et al., 2017)

• More specific to the protocol, it is used to force the exit of pathogens, toxins, transformed cells, cells infected by viruses and cells of the immune system that have interacted with all of the above, from tissue reservoirs or sanctuaries so to render them “visible” to the immune system and to the therapist who will then perform the fourth and fifth steps of this protocol

These are, respectively, the second ART screening performed after application of therapeutic ultrasounds (fourth step); and collection of samples for the molecular detection of microbes, toxins, circulating cancer cells etc. that can be detected in urine, stools, blood or serum, breath or other biological matrixes (fifth step).

We have observed that the results of the second ART, performed after application of therapeutic ultrasounds, are significantly different from those of the initial first ART. For example, signs that could be interpreted as the presence of pathogens associated with coinfections of persistent Lyme that had not been detected by the initial first ART exam, became apparent when the second ART was performed after application of therapeutic ultrasounds, thus improving the diagnostic sensitivity of the procedure. We noticed that also the inter- and intra- variability of ART results were significantly decreased when the test was performed after application of therapeutic ultrasounds.

A major advantage of the RK Protocol consists in the possibility of validating the results obtained through the first steps using specific diagnostic methods that are based on molecular biology. In the case of persistent Lyme disease, for example, we used the urine DNA test that identifies genes pertaining to agents known to infect patients with persistent Lyme. Figure 8, shows a significant example of the results of the urine DNA test performed before and after application of therapeutic ultrasounds. The case of this patient with a history of angina that could not be causally diagnosed with the classic cardiology tests (such as stress echo etc.) is shown as a paradigmatic example representing a large majority of patients whom we have observed during the implementation of the RK Protocol at Sophia Health Institute. In this patient, urine samples were collected before and after application of therapeutic ultrasounds following the RK Protocol. In the urine sample collected before application of therapeutic ultrasounds (Fig. 8, left panel) the PCR-based DNA analysis did not reveal the presence of any pathogen among those detected by the Lyme panel even though the patient had tested positive for the presence of Bartonella species with ART. In



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addition, this patient had tested negative for the presence of antibodies against Bartonella henselae (IgG/IgM) and negative also in the Western Blot test (CDC criteria) from the IgeneX lab in the blood draw obtained the same day before the application of the RK Protocol. In other words, before the application of therapeutic ultrasounds, this patient was one of the many cases where ART provided positive results that could not be confirmed by the urine DNA test or by antibody-based blood tests. However, the DNA test performed on the urine sample collected one hour after application of therapeutic ultrasounds, clearly shows the presence of one well- identified pathogen, Bartonella bacilliformis, a well- known cause of endocarditis, commonly diagnosed only post-mortem, whose presence, albeit suggested by ART, was not evident before the application of the ultrasounds (Fig. 8, right panel).

In proceeding with the implementation of the RK Protocol, the patient responded rapidly and favorably to targeted biological treatment involving daily therapeutic ultrasound application and several anti-microbial agents.

In our opinion, these results represent a paradigm change in diagnostics and therapy. Thus, the RK Protocol, enables to reach a clear diagnosis in cases that otherwise would have been labeled as “uncertain” or “undetermined” because of the discrepancy between ART and laboratory results. Quite obviously, a diagnosis based on the concordance of ART and laboratory results

enables the therapist to implement specific therapeutic approaches that would have not been possible without the RK Protocol.

Although the RK Protocol was initially developed for persistent Lyme, its potential in other conditions ranging from cancer to neurodegenerative disorders did not escape our attention. For example, in the field of oncology the RK Protocol may serve as a tool for early diagnosis and as an instrument to increase the sensitivity of cancer cells to the therapeutic effects of a variety of approaches ranging from conventional radio- and chemotherapy to more recent forms of immunotherapy (Schwalb et al., 2016). In some aspects, the RK Protocol reminds the “shock and kill” strategy that is being pursued to eliminate HIV reservoirs that are responsible for the latency and persistence of the virus. This strategy aims at inducing HIV replication in latent viral reservoir; although this concept may appear counterintuitive since the goal of antiretroviral therapies should be to inhibit, not to stimulate, viral replication, the rationale is to make the virus “visible” to the immune system and to the antiretroviral drugs (Melkova et al., 2017). Analogously, the goal of the RK Protocol is to make pathogens, toxins, transformed cells and cells infected by viruses that are inaccessible to diagnostic and therapeutic tools, “visible” so that they can be recognized and dealt with, both by the therapist and the immune system.


Fig. 8. Example of urine DNA test for the detection of Lyme infection and co-infections123

Dietrich Klinghardt and Marco Ruggiero / American Journal of Immunology 2017, 13 (2): 114.126

DOI: 10.3844/ajisp.2017.114.126

Thus, the RK protocol may find notable applications in the diagnosis of localized infections such as Lyme carditis (Robinson et al., 2015) or infections of the brain by Babesia (Aikawa et al., 1992). In these cases, that are notoriously difficult to diagnose, the RK Protocol has the additional benefit of allowing the localization of the infectious agents without resorting to invasive procedures. To this end, the therapeutic ultrasound treatment is applied to the areas of the body where the presence of the infectious agent is suspected and, if the diagnostic hypothesis is correct, the urine DNA test would show the presence of the pathogen only after application of therapeutic ultrasounds to specific areas. For example, if the presence of Babesia became evident only after application of therapeutic ultrasounds to the brain and not before, it could be concluded that the pathogen resided in the brain and was mobilized by the pressure waves generated by the therapeutic ultrasounds.

Therefore, the RK Protocol leads to profound new possibilities: Compartmentalized microbes, hidden transformed cells, toxins and metabolites could be detected using a safe and non-invasive method. In addition to these exciting possibilities in the field of diagnostics, the RK Protocol allows optimization of the therapeutic efficacy of drugs administered parenterally; thus, application of therapeutic ultrasounds before administration of drugs specifically targeted to microbes or toxins detected by the diagnostic steps of the protocol, would significantly increase uptake and utilization of the drugs themselves with a significant improvement of their therapeutic efficacy. ART would thus have the role of narrowing down the number of diagnostic hypotheses and therapeutic options with a resulting decrease of unnecessary, expensive and time-consuming tests.


We have developed a novel protocol for the non- invasive diagnosis and treatment of persistent Lyme and other chronic conditions due to persistent infections, toxicities, neoplastic transformation or neurodegeneration. The RK Protocol offers the advantage of being safe, rapid and relatively inexpensive and it can be easily implemented in any health institution whether in the developed world or elsewhere, without the need for sophisticated and expensive instruments. This protocol aims at achieving accurate and early diagnosis, at indicating the most appropriate therapeutic intervention and at maximizing the efficacy of specific therapeutic interventions.


The Authors wish to thank Ms. Daniela Deiosso for inspiring discussion and relentless support and the therapists at the Sophia Health Institute for their precious collaboration.

Author’s Contributions

Dietrich Klinghardt: The inventor and developer of ART and performed all the diagnostic and therapeutic procedures described in this study.

Marco Ruggiero: Developed the ultrasound-based techniques described in this study, wrote the first draft of this paper, provided critical input and assisted in revising and improving the paper. He had no type of involvement in the treatment of patients.

Conflict of Interest

Dietrich Klinghardt is the inventor of ART and the founder of the Klinghardt Institute, the Klinghardt Academy, the Institut fuer Neurobiologie and the Sophia Health Institute, a private clinic. Dr. Klinghardt consults for several companies producing supplements and other remedies that, however, are not mentioned in this study. Marco Ruggiero is consultant for the company “dr. reinwald healthcare”, that provided the emulsion of chondroitin sulfate, vitamin D3 and oleic acid mentioned in this study and he is the founder and CEO of the Swiss company Silver Spring Sagl, a company that produces supplements and probiotics. None of the products of Silver Spring Sagl is mentioned in this study. Marco Ruggiero is member of the Editorial Board of The American Journal of Immunology and is waived from the Article Processing fee for this contribution; he receives no remuneration for his editorial work.


This article is original and contains unpublished material. The corresponding author confirms that the other author has read and approved the manuscript.


Aikawa, M., E. Pongponratn, T. Tegoshi, K. Nakamura and T. Nagatake et al., 1992. A study on the pathogenesis of human cerebral malaria and cerebral babesiosis. Mem. Ins. Oswaldo Cruz., 3: 297-301.

DOI: 10.1590/S0074-02761992000700051 Bocchi, L., J.J.V. Branca, S. Pacini, A. Cosentino and G. Morucci et al., 2015. Effect of ultrasounds on neurons and microglia: Cell viability and automatic analysis of cell morphology. Biomed.

Sign. Proc. Contr., 22: 44-53.

DOI: 10.1016/j.bspc.2015.06.011
Bradstreet, J.J., S. Pacini and M. Ruggiero, 2014. A new



methodology of viewing extra-axial fluid and cortical abnormalities in children with autism via transcranial ultrasonography. Front. Hum. Neurosci., 7: 934-934.

DOI: 10.3389/fnhum.2013.00934

Dietrich Klinghardt and Marco Ruggiero / American Journal of Immunology 2017, 13 (2): 114.126

DOI: 10.3844/ajisp.2017.114.126

Bradstreet, J.J., M. Ruggiero and S. Pacini, 2015. Commentary: Structural and functional features of central nervous system lymphatic vessels. Front. Neurosci., 9: 485-485. DOI: 10.3389/fnins.2015.00485

Bui, L., A. Aleid, A. Alassaf, O.C. Wilson and C.B. Raub et al., 2017. Development of a custom biological scaffold for investigating ultrasound- mediated intracellular delivery. Mater. Sci. Eng. C, 70: 461-470. DOI: 10.1016/j.msec.2016.09.029

Burgdorfer, W., 1991. Lyme borreliosis: Ten years after discovery of the etiologic agent, Borrelia burgdorferi. Infection, 19: 257-262.
DOI: 10.1007/BF01644963

Burmester, G.R., 1993. Lessons from Lyme arthritis. Clin. Exp. Rheumatol., 8: S23-27. PMID: 8324947

Caulfield, A.J. and B.S. Pritt, 2015. Lyme disease coinfections in the United States. Clin. Lab. Med., 35: 827-846. DOI: 10.1016/j.cll.2015.07.006

Chomel, B., 2015. Lyme disease. Rev. Sci. Tech., 34: 569-576.

Chung, M.K. and P.J. LaRiccia, 2016. How do you deactivate painful scars in your practice? Med. Acupunct., 28: 162-167.
DOI: 10.1089/acu.2016.29023.cpl

Chung, M.K. and P.J. LaRiccia, 2017. Successful integrative medicine assessment and treatment of chronic pain associated with breast surgery: A report of 3 cases. Holist. Nurs. Pract., 31: 21-29.

DOI: 10.1097/HNP.0000000000000188
Cimpianu, C.L., W. Strube, P. Falkai, U. Palm and A. Hasan, 2017. Vagus nerve stimulation in psychiatry: A systematic review of the available evidence. J. Neural Transm. (Vienna), 124: 145-158.

DOI: 10.1007/s00702-016-1642-2
Cory, T.J., T.W. Schacker, M. Stevenson and C.V. Fletcher,

2013. Overcoming pharmacologic sanctuaries. Curr. Opin. HIV AIDS, 8: 190-195.
DOI: 10.1097/COH.0b013e32835fc68a

Cosentino, A., E. Boni, S. Pacini, J. Branca and G. Morucci et al., 2015. Morphological analysis of neurons: Automatic identification of elongations. Proceedings of the 37th Annual International Conference of the IEEE Engineering in Medicine and Biology Society, Aug. 25-29, IEEE Xplore Press, pp: 8131-8134.

DOI: 10.1109/EMBC.2015.7320281
Curcio, S.R., L.P. Tria and A.L. Gucwa, 2016.

Seroprevalence of Babesia microti in individuals with Lyme disease. Vector. Borne. Zoonotic. Dis., 16: 737-743. DOI: 10.1089/vbz.2016.2020

Economo, C. and G.N. Koskinas, 1925. Die Cytoarchitektonik der Hirnrinde des Erwachsenen Menschen. 1st Edn., Springer Verlag, Wien, pp: 810.

Embers, M.E., S.W. Barthold, J.T. Borda, L. Bowers and L. Doyle et al., 2012. Persistence of Borrelia burgdorferi in rhesus macaques following antibiotic treatment of disseminated infection. PLoS One, 7: e29914-e29914.

DOI: 10.1371/journal.pone.0029914
Eshoo, M.W., S.E. Schutzer, C.D. Crowder, H.E. Carolan

and D.J. Ecker, 2013. Achieving molecular diagnostics for Lyme disease. Expert. Rev. Mol. Diagn., 13: 875-883.
DOI: 10.1586/14737159.2013.850418

Feder, H.M. Jr., B.J. Johnson, S. O’Connell, E.D. Shapiro and A.C. Steere et al., 2007. A critical appraisal of “chronic Lyme disease”. N. Engl. J. Med., 357: 1422-1430. DOI: 10.1056/NEJMra072023

Gingrich, J.B., H.J. Harlan, P.V. Perkins and J.H. Trosper, 2001. Regional disease vector ecology profile central europe armed forces pest control board Washington DC Report Date: APR.

Hodzic, E., S. Feng, K. Holden, K.J. Freet and S.W. Barthold, 2008. Persistence of Borrelia burgdorferi following antibiotic treatment in mice. Antimicrob. Agents Chemother., 52: 1728-1736. DOI: 10.1128/AAC.01050-07

Halperin, J.J., 2015. Chronic Lyme disease: Misconceptions and challenges for patient management. Infect. Drug Resist., 8: 119-128.
DOI: 10.2147/IDR.S66739

Hameroff, S., M. Trakas, C. Duffield, E. Annabi and M.B. Gerace et al., 2013. Transcranial Ultrasound (TUS) effects on mental states: A pilot study. Brain Stimul., 6: 409-415. DOI: 10.1016/j.brs.2012.05.002

Jacobs, G.E., T.L. Franks and P.G. Gilman, 1984. Diagnosis of thyroid dysfunction: Applied kinesiology compared to clinical observations and laboratory tests. J. Manipulative Physiol. Ther., 7: 99-104. PMID: 6747487

Jensen, A.M., R.J. Stevens and A.J. Burls, 2016. Estimating the accuracy of muscle response testing: two randomised-order blinded studies. BMC Complement. Altern. Med., 16: 492-492.

DOI: 10.1186/s12906-016-1416-2
Jia, L., Y. Wang, J. Chen and W. Chen, 2016. Efficacy

of focused low-intensity pulsed ultrasound therapy for the management of knee osteoarthritis: A randomized, double blind, placebo-controlled trial. Sci. Rep., 6: 35453-35453. DOI: 10.1038/srep35453

Leinenga, G., C. Langton, R. Nisbet and J. Götz, 2016. Ultrasound treatment of neurological diseases– current and emerging applications. Nat. Rev. Neurol., 12: 161-174.

DOI: 10.1038/nrneurol.2016.13
Melkova, Z., P. Shankaran, M. Madlenakova and

J. Bodor, 2017. Current views on HIV-1 latency, persistence and cure. Folia Microbiol. (Praha), 62: 73-87. DOI: 10.1007/s12223-016-0474-7



Dietrich Klinghardt and Marco Ruggiero / American Journal of Immunology 2017, 13 (2): 114.126

DOI: 10.3844/ajisp.2017.114.126

Murgia, R. and M. Cinco, 2004. Induction of cystic forms by different stress conditions in Borrelia burgdorferi. APMIS, 112: 57-62.
DOI: 10.1111/j.1600-0463.2004.apm1120110.x

Neren, D., M.D. Johnson, W. Legon, S.P. Bachour and G. Ling et al., 2016. Vagus nerve stimulation and other neuromodulation methods for treatment of traumatic brain injury. Neurocrit. Care, 24: 308-319. DOI: 10.1007/s12028-015-0203-0

Omura, Y., 1981. New simple early diagnostic methods using Omura’s “Bi-Digital O-Ring Dysfunction Localization Method” and acupuncture organ representation points and their applications to the “drug and food compatibility test” for individual organs and to auricular diagnosis of internal organs- -part I. Acupunct. Electrother. Res., 6: 239-254. DOI: 10.3727/036012981816952199

Raper, D., A. Louveau and J. Kipnis, 2016. How do meningeal lymphatic vessels drain the CNS? Trends Neurosci., 39: 581-586.
DOI: 10.1016/j.tins.2016.07.001

Rebman, A.W., J.N. Aucott, E.R. Weinstein, K.T. Bechtold and K.C. Smith et al., 2015. Living in limbo: Contested narratives of patients with chronic symptoms following lyme disease. Qual. Health Res. PMID: 26631681

Robinson, M.L., T. Kobayashi, Y. Higgins, H. Calkins and M.T. Melia, 2015. Lyme carditis. Infect. Dis. Clin. North Am., 29: 255-268.
DOI: 10.1016/j.idc.2015.02.003

Ruggiero, M., S. Magherini, M.G. Fiore, B. Chiarelli and G. Morucci et al., 2013. Transcranial sonography: A technique for the study of the temporal lobes of the human and non-human primate brain. Ital. J. Anat. Embryol., 118: 241-255. PMID: 24640587

Ruggiero, M., E. Ward, R. Smith, J.J. Branca and D. Noakes et al., 2014. Oleic Acid, deglycosylated vitamin D-binding protein, nitric oxide: A molecular triad made lethal to cancer. Anticancer Res., 34: 3569-3578. PMID: 24982371

Ruggiero, M., 2016. Fecal microbiota transplantation and the brain microbiota in neurological diseases. Clin. Endosc., 49: 579-579.
DOI: 10.5946/ce.2016.098

Schwalb, M., M. Taubmann, S. Hines, H. Reinwald and M. Ruggiero, 2016. Clinical observation of a novel, complementary, immunotherapeutic approach based on ketogenic diet, chondroitin sulfate, Vitamin D3, oleic acid and a fermented milk and colostrum product. Am. J. Immunol., 12: 91-98.

DOI: 10.3844/ajisp.2016.91.98
Shapiro, E.D., 2014. Clinical practice: Lyme disease. N.

Engl. J. Med., 370: 1724-1731.

DOI: 10.1056/NEJMcp1314325
Sharma, B., A.V. Brown, N.E. Matluck, L.T. Hu and

K. Lewis, 2015. Borrelia burgdorferi, the causative agent of Lyme disease, forms drug-tolerant persister cells. Antimicrob. Agents Chemother., 59: 4616-4624. DOI: 10.1128/AAC.00864-15

Straubinger, R.K., B.A. Summers, Y.F. Chang and M.J. Appel, 1997. Persistence of Borrelia burgdorferi in experimentally infected dogs after antibiotic treatment. J. Clin. Microbiol., 35: 111-116. PMID: 8968890

Straubinger, R.K., A.F. Straubinger, B.A. Summers and R.H. Jacobson, 2000. Status of Borrelia burgdorferi infection after antibiotic treatment and the effects of corticosteroids: An experimental study. J. Infect. Dis., 181: 1069-1081.

DOI: 10.1086/315340
Tarasoff-Conway, J.M., R.O. Carare, R.S. Osorio,

L. Glodzik and T. Butler et al., 2015. Clearance systems in the brain-implications for Alzheimer disease. Nat. Rev. Neurol., 11: 457-470.
DOI: 10.1038/nrneurol.2015.119

Yrjänäinen, H., J. Hytönen, P. Hartiala, J. Oksi and M.K. Viljanen, 2010. Persistence of borrelial DNA in the joints of Borrelia burgdorferi-infected mice after ceftriaxone treatment. APMIS, 118: 665-673. DOI: 10.1111/j.1600-0463.2010.02615.x



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This is the first published clinical case of cancer: lifted with imuno and Bravo in 3 months.



Original Research Paper

Use of an Extremely Biodiverse Probiotic and a Supplement Based on Microbial Chondroitin Sulfate is Associated with a Significant Decrease of Serum Free Kappa Light Chains as well as a Trend Toward Normalization of Kappa/Lambda Ratio and of Plasma Cell Bone Marrow Infiltration in a Case of Multiple Myeloma

1Nicola Antonucci, 2Stefania Pacini and 2Marco Ruggiero1Biomedical Centre for Autism Research and Treatment, Bari, Italy

2Silver Spring Sagl, Arzo-Mendrisio, Switzerland

American Journal of Immunology

Article history

Received: 27-05-2019 Revised: 13-06-2019 Accepted: 18-06-2019

Corresponding Author: Marco Ruggiero
Silver Spring Sagl, Via Raimondo Rossi 24, Arzo- Mendrisio 6864, Switzerland


Multiple myeloma is a neoplasia of plasma cells present in roughly 3-4% of the population over the age of 50 years and characterized by a cytogenetically heterogeneous plasma cell population. Multiple myeloma is considered an incurable hematologic neoplasia often requiring multiple sequential therapeutic regimens that have to be constantly tailored to the changing patient’s conditions (Rajkumar et al., 2016). The standard of care for patients over 65 years and/or patients with serious concurrent chronic diseases is represented by chemotherapy with prednisone and melphalan or, more recently, with proteasome inhibitors (Hungria et al., 2019). An obstacle to chemotherapy of multiple myeloma is represented by renal insufficiency, a condition that is common in these patients and can often limit the choice of therapy (Mikhael et al., 2018).

Here we describe the case of a 71-year-old woman with multiple myeloma associated with severe chronic renal insufficiency who, since October 2018, assumed a novel, extremely biodiverse, probiotic and a

Abstract: We describe the case of a 71-year-old woman with multiple myeloma associated with severe chronic renal insufficiency who, since October 2018, assumed a novel, extremely biodiverse, probiotic and a supplement based on microbial chondroitin sulfate, vitamin D3 and ultrapure phosphatidylcholine; such a nutritional regimen was associated with a dramatic decrease of serum free Kappa light chains (from 2,190 to a minimum of 119.40 mg/L) as well as a trend toward normalization of Kappa/Lambda ratio (from 187 to a minimum of 3.16) and of plasma cell bone marrow infiltration (from 20 to 5%). supplement based on microbial chondroitin sulfate, vitamin D3 and ultrapure phosphatidylcholine; such a nutritional regimen was associated with a dramatic decrease of serum free Kappa light chains as well as a trend toward normalization of Kappa/Lambda ratio and of plasma cell bone marrow infiltration.

Case Presentation

The patient, a woman born in 1948, had been diagnosed with monoclonal gammopathy in 2010; in 2012 she developed acute renal insufficiency treated with dialysis and plasmapheresis. This event was followed by deep venous thrombosis with occlusion of the iliac- femoralis and femoralis-popliteal axes and systemic hypertension. Chronic renal insufficiency with frequent episodes of acute relapses resulted in extensive glomerular-tubular damage as evidenced by urinary protein electrophoresis; a renal biopsy documented advanced glomerulosclerosis with severe interstitial damage. In October 2018, the patient was admitted at the Oncohematological Department of the San Matteo hospital of Pavia, Italy; the document released by the hospital on October 29, 2018, reports that bone marrow biopsy of the posterior iliac crest confirmed a diagnosis of multiple myeloma with monotypical plasma cell bone marrow infiltration accounting for 20% of total cells. Study of serum free light chains showed significant alteration of the Kappa/Lambda ratio (free Kappa: 2,190 mg/L. Free Lambda: 11.7 mg/L. Ratio Kappa/Lambda: 187) that was consistent with the severity of the oncohematological disease according to the updated criteria for the diagnosis of multiple myeloma (Rajkumaret al., 2014). Following hospitalization in October 2018, the patient opted for a complementary approach to her condition consisting in consumption of 50 mL, twice a day, of an extremely biodiverse probiotic drink obtained by fermentation of bovine milk and colostrum; in addition, the Medical Doctor supervising the patient, in October 2018 begun administering 0.2 mL per week of a novel immune-modulating supplement based on microbial chondroitin sulfate, vitamin D3 and ultrapure phosphatidylcholine (Ruggiero, 2018). A laboratory exam performed three months later (January 10, 2019) demonstrated a 95% decrease of serum free Kappa light chains that went from 2,190 mg/L (October 2018) to 119.40 mg/L. Also the ratio Kappa/Lambda showed a trend toward normalization decreasing from 187 (October 2018) to 11.10 (January 2019). Consistent with the significant improvement of the oncohematological picture, also the renal function improved, albeit to a lesser extent. Creatinine decreased from 1.76 mg/dL (October 2018) to 1.61 mg/dL (January 2019); glomerular filtrate increased from 28 mL/min (October 2018) to 34 mL/min (January 2019). Two months later (March 1, 2019) the patient was admitted to the Hospital of Eboli, Italy, for a trauma to her left leg due to an accidental fall at home; the laboratory exams performed on that occasion demonstrated further improvement of the oncohematological condition with the ratio Kappa/Lambda decreased to 3.5 that is very close to the upper limit of normal values (2.93). A further laboratory exam performed about two months later (May 24, 2019) at the University Hospital San Giovanni di Dio e Ruggi d’Aragona, Salerno, Italy, confirmed the continuing descending trend of the Kappa/Lambda ratio that was 3.16. Consistent with these results, a bone marrow aspiration at the sternum performed on April 26, 2019 at the San Giuseppe Moscati Hospital, Avellino, Italy, evidenced plasma cell infiltration of 5% that is a very significant reduction in comparison to the results observed before implementing the nutritional approach described above; thus, plasma cell bone marrow infiltration was 20% in October 2018. The report from the San Giuseppe Moscati Hospital was consistent with a quasi-normal

condition as it read: “Bone marrow aspiration. Slightly reduced bone density. Specimen very rich in cells and tiny lumps. Parenchymal erythroblasts and granulocytes are normal and in normal stage of maturation. Number of lymphocytes is normal. Very slight (in Italian, lievissimo) increase of plasma cells that are present in a percentage of about 5%; some plasma cell is binucleate. Megakaryocytes are numerous and most of them in active terminal thrombopoiesis.” (literal translation from Italian).


A number of preclinical and clinical trials highlights the importance of the microbiota in cancer and there is general consensus that modulation of the microbiota plays a fundamental role in determining the response of cancer to therapies (Gopalakrishnan et al., 2018). The observation described in this case supports the hypothesis that a combination of a microbiota-based approach with immune-modulating strategies may prove effective in oncohematological diseases. It is worth noticing that the milk and colostrum fermented drink assumed by the patient here described represents a novel form of probiotic because of its extreme biodiversity; ingredients, microbial composition and nutritional characteristics of this drink are described in Table 1. A microbiome assay performed with a chip covered in DNA sequences that are specific to target organisms and is able to detect a total of approximately 12,000 species (Thissen et al., 2019), revealed the presence of hundreds of different microorganisms, thus showing an extremely high degree of biodiversity that included plasmids and phages with the latter being heralded as the tools for the future of immunotherapy (Rehman et al., 2019). It is known that phages are endowed with anticancer activity and it was demonstrated that they inhibit the growth of experimental melanoma and lung cancer and also inhibit the formation of lung metastases (Dabrowska et al., 2004). In addition, phages are responsible for activation of tumoricidal, classically activated, M1 macrophages and it was demonstrated that, following cancer treatment with phages, the M2-polarized tumor microenvironment switched toward an M1-polarized milieu (Eriksson et al., 2009). At the molecular level, an example highlighting the role of the phages is represented by the presence ofLactococcus lactis phage ul36 (Siphoviridae) in the probiotic drink assumed by the patient. As demonstrated since 2008, this microorganism encodes a single-strand annealing protein (Sak) that is homologous to the human recombination protein RAD52, a protein that plays crucial roles in DNA repair and genome stability (Ploquin et al., 2008).



Table 1: Characteristics of the probiotic


Description of the product Ingredients, colostrum Ingredients, microbial blends

Nutritional information

Product based on bovine colostrum and live probiotics for home preparation of fermented milk.
Freeze-dried, pasteurized bovine colostrum. 5 grams per liter of final product.
Blend of lyophilized kefir grains containing live cultures (including Streptococcus thermophilus and Lactobacillus delbrueckii, subspecie bulgaricus) and yeasts.
Blend of Bifidobacteria: Bifidobacterium infantis (25%); Bifidobacterium bifidum (25%); Bifidobacterium lactis (25%); Bifidobacterium longum (25%).
Blend of lyophilized micro-encapsulated live cultures; Bifidobacterium infantis (12.5%); Bifidobacterium bifidum (12.5%); Bifidobacterium lactis (12.5%); Lactobacillus salivarius (12.5%); Lactobacillus acidophilus (12.5%); Lactobacillus casei, subspecie paracasei (12.5%); Lactobacillus rhamnosus (12.5%); Lactococcus lactis, subspecie cremoris (12.5%). Coating agents for this blend (micro-encapsulation); mono- and diglycerides of fatty acids. Calories: 4 Kcal. Proteins: 0.28 grams. Fats: 0.28 grams. Carbohydrates: 0.22 grams. These values refer to the ingredients listed above in the measure required to prepare 1 liter of final product.


The ingredients listed above are used by the final utilizer to ferment bovine milk at room temperature for 24 h and prepare a probiotic drink. The microbiome assay described in the text refers to the product of fermentation and not to the individual ingredients listed above. The microbiome assay was performed by an independent laboratory on the fermented probiotic drink that was lyophilized prior to the assay. We hypothesize that the extreme biodiversity mentioned in the text is due to the process of fermentation as well as to the complex microflora of the kefir grains.

From an evolutionary perspective, however, it is the human RAD52 protein that has evolved from the phage Sak protein, thus further emphasizing the role of phage-associated genetic information in maintaining DNA stability and, in a broader sense, human health.

It is tempting to speculate that the wide-range of effects on the immune system deriving from the extreme biodiversity of the probiotic drink assumed by the patient was synergistic with the effects of the supplement based on microbial chondroitin sulfate, vitamin D3 and ultrapure phosphatidylcholine as they are described in two recent papers (Ruggiero, 2018; Ruggiero and Pacini, 2018). Such a synergism may well be at work at the molecular level, for example by integrating different mechanisms responsible for DNA repair that counteract DNA instability, a common trait of cancer and other conditions. On one hand, the Sak/RAD52 pathway of the probiotic targets single-stranded DNAs preferentially over double-stranded DNAs and promotes the renaturation of long complementary single-stranded DNAs (Ploquin et al., 2008); on the other hand, the vitamin D3 of the supplement promotes expression of the DNA repair genes RAD50 and ATM (Ataxia Telangiectasia Mutated), that play a key role in mediating the signaling responses to double-strand DNA breaks (Ting et al., 2012). The convergence of these pathways maximizes the ability of cells to restore single- and double-strand DNA breaks thus efficiently counteracting a variety of DNA damages that are considered responsible for cancer onset and progression. It is also worth emphasizing the synergism between the phages in the probiotic and the phosphatidylcholine in the supplement in promoting and stabilizing a strong M1 phenotype in macrophage polarization (Qin et al., 2014).

Since this clinical case report is an open-label, non- controlled, retrospective analysis, caution must be exercised when ascribing cause and effect to any

treatment outcome. In addition, as of June 2019, the patient is assuming the following drugs in the context of chronic renal disease management; prednisone, 5 mg/die; omeprazole, 20 mg/die; folic acid, 5 mg/die; febuxostat, 80 mg/die; telmisartan, 20 mg when needed; warfarin 5 mg administered according to the patient’s INR. The patient did not report side effects associated with the nutritional approaches described above assumed in conjunction with her current therapy. Although prednisone may have contributed to the observed results as it is used in the therapy of multiple myeloma, it is worth noticing that the patient had been assuming this drug alongside others for a long time. In addition, the dose for the management of multiple myeloma is ten times higher than that assumed by the patient (Berenson et al., 2002). Despite these limitations, it appears that the association between the probiotic and the supplement and the improvements demonstrated by objective laboratory data was robust. It is well accepted that case reports are constituents of evidence-based medicine and informed practice (Juyal et al., 2013) and we are convinced that this novel and unusual clinical observation may lead to subsequent research advances in the field of complementary immunotherapy of cancer.


The Authors wish to thank the patient whose case is described here and her relatives for their priceless collaboration.

Conflict of Interest

Nicola Antonucci is the founder of the Biomedical Centre for Autism Research and Treatment, a private clinic. Marco Ruggiero is the inventor of a number of supplements and, together with Stefania Pacini, developed the probiotic (Bravo, Silver Spring Sagl, ) and the supplement (imuno, imuno Corporation, Vanuatu) used in this study. Both the probiotic and the supplement were regularly purchased and paid for by the patient. Neither Dr. Ruggiero, nor Dr. Pacini, had any prior knowledge of the strategies being implemented nor of the details of the patient whose clinical outcomes were analyzed and described in this study. Marco Ruggiero is member of the Editorial Board of The American Journal of Immunology and is waived from the Article Processing fee for this contribution; he receives no remuneration for his editorial work.

Authors’ Contribution

Nicola Antonucci: Performed the therapeutic procedures described in this study and communicated the clinical and laboratory data of the patient.

Marco Ruggiero and Stefania Pacini: Wrote the first draft of this paper, provided critical input and assisted in revising and improving the paper.


No information in this paper is presented by the authors as medical advice. Caregivers, researchers and interested parties should research all information given. Beginning any significant biomedical or other interventions that may impact physiology or making changes to an established regimen should be discussed with the patient’s physician in advance. Standard of care for each pathology must be followed as well as rules and regulations established by Health Authorities of each Country.


Rajkumar, S.V., 2016. Myeloma today: Disease definitions and treatment advances. Am. J. Hematol., 1: 90-100. DOI: 10.1002/ajh.24236

Eriksson, F., P. Tsagozis and K. Lundberg, R. Parsa and S.M. Mangsbo et al., 2009. Tumor-specific bacteriophages induce tumor destruction through activation of tumor-associated macrophages. J. Immunol., 182: 3105-3111.

DOI: 10.4049/jimmunol.0800224
Ploquin, M., A. Bransi and E.R. Paquet, A.Z. Stasiak and

A. Stasiak et al., 2008. Functional and structural basis for a bacteriophage homolog of human RAD52. Curr. Biol., 18: 1142-6.
DOI: 10.1016/j.cub.2008.06.071

Ruggiero, M. and S.A. Pacini, 2018. Novel potential adjuvant for cancer vaccines. Madridge J. Vaccines, 2: 57-1. DOI: 10.18689/mjv-1000112

Ting, H.J., S. Yasmin-Karim, S.J. Yan, J.W. Hsu and T.H. Lin et al., 2012. A positive feedback signaling loop between ATM and the vitamin D receptor is critical for cancer chemoprevention by vitamin D. Cancer Res., 72: 958-68.

DOI: 10.1158/0008-5472.CAN-11-0042
Qin, X., C. Qiu and L. Zhao, 2014. Lysophosphatidylcholine perpetuates macrophage polarization toward classically activated phenotype

in inflammation. Cell Immunol., 289: 185-90.

DOI: 10.1016/j.cellimm.2014.04.01015
Juyal, D., S. Thaledi and V. Thawani, 2013. Thawani writing patient case reports for publication. Educ.

Health, 26: 126-9. DOI: 10.4103/1357-6283.120707 Hungria, V.T.M., E.Q. Crusoé, R.I. Bittencourt, R.P. Magalhães and J.N. Sobrinho et al., 2019. New proteasome inhibitors in the treatment of multiple myeloma. Hematol. Transfus Cell Ther., 1: 76-83.

DOI: 10.1016/j.htct.2018.07.003
Mikhael, J., J. Manola, A.C. Dueck, S. Hayman and K.

Oettel et al., 2018. Lenalidomide and dexamethasone in patients with relapsed multiple myeloma and impaired renal function: PrE1003, a PrECOG study. Blood Cancer J.

DOI: 10.1038/s41408-018-0110-7
Rajkumar, S.V., M.A. Dimopoulos, A. Palumbo, J.

Blade and G. Merlini et al., 2014. International myeloma working group updated criteria for the diagnosis of multiple myeloma. Lancet Oncol,. 15: e538-48. DOI: 10.1016/S1470-2045(14)70442-5

Ruggiero, M., 2018. Rationale for the design of a novel tool for immunotherapy based on an emulsion of glycosaminoglycan. Integr. Cancer Sci. Therap, 5: 1-5. DOI: 10.15761/ICST.1000285

Gopalakrishnan, V., B.A. Helmink, C.N. Spencer, A. Reuben and J.A. Wargo et al., 2018. The Influence of the gut microbiome on cancer, immunity and cancer immunotherapy. Cancer Cell, 33: 570-580. DOI: 10.1016/j.ccell.2018.03.015

Thissen, J.B., N.A. Be, K. McLoughlin, S. Gardner and P.G. Rack et al., 2019. Axiom microbiome array, the next generation microarray for high-throughput pathogen and microbiome analysis. PLoS ONE. DOI: 10.1371/journal.pone.0212045

Rehman, S., Z. Ali, M. Khan, N. Bostan and S. Naseem et al., 2019. The dawn of phage therapy. Rev. Med. Virol. DOI: 10.1002/rmv.2041

Dabrowska, K., A. Opolski, J. Wietrzyk, K. Switala- Jelen and J. Boratynski et al., 2004. Antitumor activity of bacteriophages in murine experimental cancer models caused possibly by inhibition of 3 integrin signaling pathway. Acta Virol., 48: 241-248. PMID: 15745047



Nicola Antonucci et al. / American Journal of Immunology 2019, ■■ (■): ■■■.■■■DOI: 10.3844/ajisp.2019.■■■.■■■

Berenson, J.R., J.J. Crowley, T.M. Grogan, J. Zangmeister and A.D. Briggs et al., 2002. Maintenance therapy with alternate-day prednisone improves survival in multiple myeloma patients. Blood, 99: 3163-8. DOI: 10.1182/blood.V99.9.3163

Accessory Material

Clinical and laboratory records are archived at the Biomedical Centre for Autism Research and Treatment, Bari. Italy. Since this is a low-number case report that does not produce generalizable knowledge, nor an investigation of an FDA regulated product, Institutional Review Board (IRB) review is not required for this activity (John Hopkins Medicine. 102.3 Organization Policy on Single Case Reports and Case Series. oard/guidelines_policies/organization_policies/ ml. Web page accessed June 10, 2019).



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The Alpha Lipoic Acid Breakthrough

The Alpha Lipoic Acid Breakthrough: The Superb Antioxidant That May Slow Aging, Repair Liver Damage, and Reduce the Risk of Cancer, Heart Disease, and Diabetes                                                                                                                                      By Burt Berkson, M.D., Ph.D.

“Alpha lipoic acid is probably the most potent .  .  . antioxidant known to man.                       — World News Tonight with Peter Jennings

The Alpha Lipoic Acid Breakthrough Burt Berkson, MD Ph.D.


National Bestseller:

The Complete Nutritional Program to Prevent and Reverse Insulin Resistance SYNDROME X : Use the Glycemic Index to Increase your Glucose Tolerance  By Jack Challem, Burton Berkson, M.D., Melissa Diane Smith

Lose Weight * Stop Heart Disease and Diabetes * Clear up Mental Fuzziness * Increase Energy

The Complete Nutritional Program to Prevent and Reverse Insulin Resistance Syndrome X


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Postbiotic Metabolites: The New Frontier in Microbiome Science

KIC Document 0001

by Ross Pelton, RPH, Ph.D., CCN

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Mimi’s Kitchen Club – Local Pick-up

Mimi’s Kitchen Club is a network of people who make prepared Bravo Yogurt and in some cases, other products will be available as well.  Purchases must be made online.   The full rate with postage will be charged.  If the product is not picked up, it will be mailed.  If it is picked up, your postage will be refunded.

Each prepared yogurt is made to order.  In cases where a product order needs to be picked up, please make arrangements for the handoff ahead of time.

Please call the Bravo Coop order desk at 321-747-5517 for more information or to make arrangements.

Central Florida:

  • Winter Park Farmer’s Market – Monday Nights
  • Orlando/Winter Park – Every other Thursday morning.   Please make arrangements for pick up and make the purchase online
  • Christmas – Sunday afternoons at the dairy farm (pls call to confirm availability & timing)
  • Titusville – By appointment weekdays  8 AM – 4 PM (pls call to confirm availability)

To confirm arrangements for pick up or for more info at Bravo Coop order desk –            Call 321 747 5517

Are you someone who makes Bravo Prepared Yogurt or others? Please consider being part of Mimi’s Kitchen Club and helping others.


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Experiences with imuno – a benefit to the immune system

Imuno™ – The Solution

This is the product and where you can buy it.

A year and a half ago, I took 3 high dose imuno suppositories over the course of a week. At the end of the week, all of a sudden, the skin glands of my face had tiny pockets of pus come out of my pores.  An hour later gunmetal gray powder came out of my pours as well.  I had to scrub it with a face cloth.  By taking too much at one time, it popped through the pressure system of the cranial nerves and “blew a bit of dust through the pores”.

My body calmed down and the heavy metals moved out in an orderly fashion with no event or healing crisis.  I have never had so many heavy metals move at once.  I was delighted.  For me, that means, I got rid of it and it won’t hurt me when I am older.  But on the ground at that moment, it was as if a train going down my spine filled with train cars of heavy metal.  It went to the back of the sacrum and then and out through the colon.

This migration was not uncomfortable.  But I was also not at “full function” with mental acuity.   I simply sat there and waited for the train to pass.  When it did, my brain was bright and clear!  The heavy metal was aluminum and apparently came out from the brain stem.  I rejoiced.   I am grateful to get the metals out.

One of my customers was able to move out of a devastating condition in a few months.  When I got my own imuno, I called him to see what he was doing.   He told me that he was taking 11 drops of Rerum (this was a while ago and would be like 8 drops of imuno) twice per week in a nebulizer.  That seemed like a very small price to pay to have disease removed.  I asked my husband to do it with me.

We got out our brand new nebulizer, the vial of imuno, and a syringe.  We took the product out of the vial with a syringe to keep it sterile.  We counted out the number of drops that we wanted from the syringe.  I put some tape over some side holes of the mouth piece.  I used it under my nose.  We put the drops straight into the cup of the nebulizer, turned it on, and took a few inhalations until the liquid was gone.

The nebulizer is like a fish bubbler.  It just pushes air through tubes.  So when we want to inhale, we turn the motor on and keep it on until we are done inhaling then turn it off. I usually hold my breath until I feel the exchange is complete then breath out and take some normal breaths to get comfortable.  After a rest, we take another breath from the nebulizer.  I usually let all the air out and really empty my lungs yoga style before inhaling the imuno so that I can start through the whole breath from the beginning.  It didn’t take many more than 4 breaths all total!

The treatment left a dazzling clean spot the size of a peach in the center of my bronchial tubes as well as another ball shape inside my sinuses.  It is beautifully clean and amazing feeling!  I could breathe and exchange oxygen so maybe 80% better for a little while until my breath went back into a slower more sedentary pattern!   I began to investigate why I felt so good.   I  exercise so it wasn’t about being in shape or getting more oxygen into my body.  It was something else.

The second dose was 3 days later and it was even more telling!  I breathed in the imuno, it sailed right through the nice clean part, and spread out into the next adjacent area which became all sparkling clean!   It was like blowing up the balloon, the clean part was now bigger by 1/3 of an inch more in both my lungs and my sinuses.

I enjoyed that clean shiny feeling for a few days.  Then, three days later, I was ready for some more.  I took another dose with the same result. Wow!  My head was much clearer than I ever saw it!

The next dose began to challenge my willingness to heal.  I felt a little tired.  It didn’t feel uncomfortable but I was working!   So I waited until the 4th day to see if I could change it up.  This is when I realized that the crisp clean line between clean and whatever I was before, got a tiny bit fuzzy.  I knew that I preferred the clean hands down!  I knew that I wanted to go back to 3 days.  So I did.

Soon, the clean line got to my heart and to my diaphragm, and then my liver.  It was still cleaning, but now I could feel it kind of working harder.  Not quite grinding but something like that and the stream of waste was a bit harder to manage to get it out.  Not uncomfortable, not overwhelming because the Bravo Yogurt had cleaned me out so well.

Here is when I realized it was like driving a manual transmission.  I no longer could drive in first gear.  I had to take a little less power, so I reduced the drops to 5 and kept my 3 day pace.  This gave me more energy to use in my life and limited the resources I had to spend to move old viruses and toxins in the liver.  I went to sleep dreaming about having a majorly clean body.  I was delighted because I could function.

The next phase was much like going from 2nd to 3rd gear and eventually to 4th gear.  This was more like balancing the amount of energy I had in my day and how much I could give to the ongoing imuno process.  I had to have resources to move the stuff out, keep it nice and clean and then continue to move out the waste from the areas it was released into.  As long as I didn’t get too much going that I had to “wait for the train”, I was fine!

Although it seems counter intuitive, I had to take less and less as I went on.  I just didn’t need it because I needed my energy to move the toxins out.  It was working, I just can’t push it.  That’s the wrong thing to do.  A gentle stream or torrential flood over the banks?  I take the happy and gentle cause that’s what I want in my life.  Mind you, as the balloon got bigger, the size of it’sexpansion increased.  Atthe beginning, an increase yeildedmuch less than it does 6 weeks into this.  Much more toxic release was created, and I wanted to keep on 3 days.  My drops lowered and lowered!  My body got more and more dazzling.

There are times when my shiny new friend was grinding through the removal of denser material.  I am not going to stop that action!!!  It is the best thing that has ever happened to me because I don’t have to support this.  The microbiome and the imuno in the inate immune system are supporting it and the use of those together makes sure that everything is good.  I just needed to know how to pay attention to how much I need.

PS.  taking breaks wasn’t as fun as I thought.  It’s like being a really good athlete, you don’t really want to stop stretching out and working the body.  You can slow down but not stop.  So I just kept taking less and less.  When I get to a minimal dose, I went to the suppositories, then back to the nebulizer, then to the salve, then back.

I am really happy.  My life got so much easier to manage.  I had to keep checking to make sure everything was alright.  I might have taken more naps, but everything was getting better and better while things got easier and easier.

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4 Ways to Take Bravo!

4 Ways to Take Bravo!

When you are considering how to best use Bravo GcMAF products, one must look at where you need it the most! Finding the area in your body that is in the most distress can also be the area that can be most benefited. However, I can easily build a case for all of these methods!

So much Bravo, so little time!

See your health practitioner and use your common sense. Don’t over-take Bravo because you will need to remove the toxins it cleans. We highly recommend using Bravo Protocol with all of the methods below and not to use more Bravo than is comfortable over the long term.

Sure, you can have 8 cleaning crews in your home working day and night but it would exhaust you! Sure, you can stand at the pitching machine, responding to the pitches with your helmet and bat but would 5 pitching machines pointed at you be as much fun as one? Limit your use of Bravo to the amount that is fun, easy to manage and don’t worry; it will all get done! Like exercise, yoga or music lessons, you will grow better at taking more Bravo bit by bit.

The 4 Methods of Taking Bravo

1 Yogurt 

– Slowly sipping, swishing and swallowing the yogurt assists mostly the upper part of the body! The brain, nerves, throat, thyroid, heart, lungs, esophagus, and the walls of the stomach all receive the most.

Over time, the beneficial bacteria permeates to the connective tissue, muscles, and nerves giving us comfort and the feeling that everything is contained and manageable.

Our immune system, supported by a ketogenic diet, slowly clears out the potential heat/inflammation and residual infections which may fester. The beneficial bacteria move in to cool and insulate the nerves, making our thought process clearer, deeper, and more focused. I am not a doctor! This is just what I experience while taking Bravo. Experience it for yourself over time.

I personally experienced the effect in my intestines while taking yogurt, so I know the yogurt reaches there too. I also experienced my thought processes going from being able to handle one thing at a time to two, three and even four parts of the same analysis at once! It reminded me of being in college. (and that I wasn’t doing it anymore!) I personally gained lots of mental function from taking Bravo!

2 Capsules 

– The enteric coated Bravo Concentrate Caps speed past the stomach acid for delivery to the area of the pancreas, small intestine, liver, tubes of the digestive system, the stomach and even the spleen and kidneys getting it coated, cleaned, and brightened more quickly than yogurt alone.

The Bravo Concentrate holds the complete action and nutrition pattern of Bravo GcMAF yogurt in capsule form and is made from prepared Bravo GcMAF Yogurt which is dried at a low temperature and it is not from starter powders mixed together. It can stay stable without refrigeration making it great for travel.

For me, I feel brighter! It is like the lights “turned on” in that area and I had no idea how much my pancreas needed attention and how quickly it cleared up after years of not tolerating any sugars!

3 Suppositories – 

Taking a suppository brings Bravo GcMAF action to the colon, bladder and reproductive system which can harbor low-grade infection. Bravo Coop offers different strengths in the suppositories to help you find the right daily level for you.

For me, the suppositories seemed to transform this area into a lighter and more breathable feeling. Also, I felt the whole colon, duodenum, stomach, esophagus, and soft palette feel more connected and awake. I even experienced an increase in being able to taste my food!

I went from not liking the idea of suppository use, to not wanting to miss this wonderful experience!

4 Topically on the Skin – 

Have you tried this yet? Take a small amount (1/2 tsp) of Bravo Yogurt and apply a thin layer to the skin of your face and neck and let it dry. Soon your skin will feel like a baby’s skin! It feels hydrated, full, strong and happy. I experienced fewer wrinkles and that it was easy to do!

The skin is the primary defense of the body. It defends against bad microbes among other things. The skin is also the brain of the body, according to Bruce Lipton, a Stanford cell biologist, in The Biology of Belief. Immune support in this area allows us to stay in touch with reality and defend ourselves effectively.

I have always “fed” my skin naturally, with aloe vera, fresh seed oils, and coconut oil. Now that I have Bravo to use as a natural emollient lotion, I get so disappointed when I forget this step! It is such a benefit!

How to mix applications of Bravo – With all 4 types of applications, the dosage approach stays the same.

– Begin slowly with a very small amount (approximately 1/4 tsp) x 2 times per day for 1 week and then build up each week a bit more, as in 1 tsp, 1 TBS, 2 TBS. – 2 x per day. etc. Dr. Ruggiero suggests not taking more than 1/2 cup per day, however, I take far less. If you are mixing applications, you still stay under this guideline. Perhaps one dose is one application and another dose is another, or you start in your area of concern and branch out to other applications when you are taking more.

– After the first month, follow your doctor’s advice or if alone, use your body’s lead but you want your body to feel great.

– If you are taking too much, you may feel overwhelmed, too much to respond to or too busy.

– If you take too little or are not eating well enough, you may not feel anything. You may not feel any benefit, energy, or “brightening”. If this is happening, clean up the diet, take vitamin D, hold to your startup for 1 month, then you may consider taking tiny increments until you feel great and then hold at that level.

– When planning to mix different Bravo applications, common sense prevails. If you are currently taking “x” x 2, for example, stay within your daily dosage even if you are applying it several different ways. Any combination as long as it doesn’t exceed the daily amount.

– The start-up amounts for both capsules and suppositories are for sale in “start-up kits”. This makes it easy to increase over the first month of use for either application.

– When using capsules or suppositories, look on the Bravo Coop site for products in the application you wish to use that are similar to the level you are currently taking.





Q:  What is the equivalent dosage of the capsules?

A: The capsules have dosages that equal the yogurt’s for example . . .   1 teaspoon, 1 tablespoon, 2 tablespoons, etc.   What ever dose you take in yogurt, you can take it as a capsule instead.  if you are comfortable taking 2 TBS of yogurt, twice per day, you can take two tablespoons of yogurt, capsules or suppositories as a mix and match twice per day.  Just don’t take more than you are comfortable with.  If you can’t feel it – don’t push it!  More is not better. For example, if you are taking 2 TBS of yogurt you would want to stay at 2 TBS per dose but it won’t matter if it is yogurt, or caps.  You might take a  2 TBS capsule  and 2 tbs suppository  mix and match.

You can even get fancier and split it.  If you take 2 TBS per dose,  twice per day.  You might take split doses with different items.  such as 1 TBS yogurt and 1 TBS capsule in one dose and the same at the other dose.  Or mix it up.  a 1 TBS suppository and a 1 TBS capsule at same dose.    You are still taking 2 TBS at each dose.

Q: Would be o.k. to start with a quarter teaspoon of yogurt PLUS one capsule twice a day or whether that would be too much?

A: That would be too much.  The first month is about building up slowly.  Why?  Because the yogurt grabs onto the tissue and shimmers it and the tissue detoxifies.  Your body needs to move that toxin out.  The more you take, the more territory will be renovated.   How many cleaning crews are you comfortable with? More is not better.  Be comfortable and have easy access to happy good living.  Feel great while getting better.  If you have to work too hard, you might get discouraged and there is no reason for that.

Q: Since the capsule gets down further, and I’m trying to address my colon, maybe this would be o.k?

A  if you are trying to address the colon, why not take suppositories?  Just be careful to regulate the dosage.

Q: Do the capsules also have the GCMAF and are they the same potency as the yogurt?

A:  Neither the yogurt or the caps technically have GcMAF.

All of the above methods have the essential ingredients to create a live microbiome who then produces the GcMAF.

It is that essential microbiome that we are building and the doctor’s formula of ratios that we are protecting.

To me, it seems like we need to continue to feed and replenish it.  If I stop taking it, it slowly fades based on my diet.

I observe mostly when I travel or run out for a few days.  Our bodies are big!  It takes a lot of product to fill all the places and get between the nerves and connective tissue, etc.  I find that the yogurt is always working.  No worries about that.  Just don’t let it die like house plants and pets by not feeding it and providing what it needs.  I consider replenishing it when I take more because it has the food in it, (colostrum) and the housing (probiotics) and the cute little beneficials.

Again, my experience is that the Yogurt has it all and is readily available. It will go in and start homesteading right away.  The capsules have to rehydrate and come alive which it does but it takes a while.

I experience the capsules as a “brightening” through my solar plexis area .  (Pancreas, gall bladder, small intestines, bottom of the duodenum and diaphragm.   I experience how happy my body is to have this interaction and the fade when I don’t take the capsules.  I started the yogurt a year and a half before the capsules came out so if my gut was going to fill up, it would have already.  Taking the capsules was like turning on a new room in my body which had not seen Bravo before.

I hope this helps.  It’s an amazing feeling to be brightened!!!  I won’t stop taking the yogurt.  I also take somNon-dairy bravo at times.   I am experimenting now to measure if it reduces inflammation.

In terms of potency, the caps can have different potencies.  We don’t want to overwhelm a new area and it seems that each place I use it, it starts a new colony so the rules apply but only in terms of how it detoxifies.

Q: I’m not sure what to order.  I’ve only ever taken 2 tablespoons (one capsule before bed every night).  Do you think I’d herx too hard if I took 4?  Not sure if I should try upping the dose, or just stick to what I’m used to.  Any advice would be helpful.

A: Short answer: Your body likes 2 TBS.  I would stay with that.  You are happy with what you are doing.

Long answer: If your body wants to and you have a good sense of that from listening to your inner needs, you can take 4 TBS sometimes simply by taking 2 capsules at that time.  I found that when my immune system “opened” and filled up my body like a space of connective tissue, it did need more.  It was a short lived demand. This is an advanced immune move that took many months to get to but a good diet, lots of greens and protein, low to no carbs and lots of breath and exercise will really help to keep your body clean.  By opening up, your immune system can open up biofilms and take out the complete colony of unwanted microbes.   Then the tissue “re-orders” that area back to healthy divine tissue.  That’s when the good guys need to move it.  The Microbiome moves in.  I think it has a big “S” on its chest.

Do you have more questions?



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How do you want to keep your Bravo Yogurt culture warm?

There aren’t detailed instructions here.  Just some help with understanding the culturing.

Hi, I am Mimi.  I make at least 2-5 gallons of yogurt for sale to customers every week since the beginning of 2016 when I started to sell it.  I have been through everything I can think of in terms of making it.  I also end up using marginal products that don’t come out right.  I, therefore, have grown attentive to doing it right and have a healthy respect for what is likely to fail.

Everyone gets a bit excited to bring home Bravo Yogurt.  Some are big culturing people. others have never considered a double boiler or tasting something to see if it is done.  Yet we all need to make Bravo Yogurt.  There are as many ways to make it as there are people.

Usually, I can go over the basics with people and they understand but consistently, we need to talk about what will work to culture the yogurt.  I have settled on the best recipe for culturing through trial and error with thousands of people.  It is 93 degrees for 12 hours.  Within that is a range.  88 for more hours will work, etc.  I just don’t know what that hourly amount will be.  It must be tasted to determine.  Does it get cooler at night?  how much cooler?  on and on.

I have come up with a solid way to make the yogurt and I have come up with the description of a “taste method” to help out knowing when the culture is done when we are off roading.  Warning: don’t just pick a recipe from the internet from Bravo Yogurt.  There are some out there which are for older recipes that don’t really relate.  Try to stay current.  Have a question?  Ask me.  mimi @ bravocoop dot com.

The first part of making Bravo is the milk.  Raw, fresh and whole.  As close as it comes to the real thing.  Then we will heat it to 160 to 185 to home pasteurize it and to remove the casein which is the stress hormone complex residing inside the milk.  If the milk is pasteurized, it will still need to be heated and the skin that comes to the surface must be removed.  Don’t eat this.  It’s really all the noise from the barnyard. Once it is removed, the milk is a clean canvas and ready for the powders.

Keeping it warm is where people need to use what they have.  For example, you may not have all of these, but choose the one that works for you!!

Once you find the method you want to use, it’s important to calibrate as needed.  To calibrate, test a bowl of water with a thermometer.  If it is near 93˚ you are okay to try the good stuff.  Otherwise, water in your container and put it in your warming environment. modify as needed.  It may need to be re-adjusted with the seasons.

  1. Gas ovens have a pilot light –
    1. Modification – closer or nearer to find right temp
  2. Proofing function – an option on some electric ovens
    1. Check the temp & timing for your brand
  3. Oven light bulb (only- no heat used)  –
    1. Modification – closer or nearer to find the right temp
    2. Use a post-it “Keep the light on!  Bravo!”  That’s how I got our tag line.
  4. Instant pot  – It must have a Yogurt button
    1. Test the temperatures with thermomenter
    2. If it doesn’t have an adjust button, use the yogurt button again
  5. Yogurt machine –  – see FAQs How to Make
    1. These are factory set at 110˚ but you can use them if you lift the vessel off of the bottom (direct heat) using something 1″-2″ high to create ambiant heat.  Rack?
  6. Reptile mat from pet store
    1. Modification – closer or nearer to find the right temp or use a towel to wrap.
    2. Setting directly on this is how it is built.  Usually it is radiant heat at the right temp
    3. Not very expensive and easy to store.
  7. 40 oz Thermos King
    1. Heat your milk or your water test to 93 degrees and put the lid on – measure in 12 hours.
    2. Inexpensive, diverse uses, no electricity needed. unbreakable
    3. The downside, it has to be covered with a lid and may benefit more oxygen.

Failed projects

  • Don’t use boiling hot water suspended inside a cooler due to bacterial growth.  Use dry heat!
  • On the counter for 2-3 days, the milk could go bad before it begins to culture.  unnecessary failure has been observed
  • Dehydrator ( I have trouble with the noise)

Advanced projects

  1. Drift after “culture” is established
    1. Culture the yogurt for 12 hours at 93˚ to establish the culture
      1. You can use a Christmas tree light  timer at the plug to turn it off at 12 hours.
    2. Let the temperature drift to room temp for up to 5 hours.
      1. Since all the organisms have different culture rates, this makes them happy.
      2. It also lets you sleep or get home from work, etc.
    3. Make it easy for yourself  – at 7 pm (dinner) and collect it at 7 am (breakfast)
    4. Use an internal timer like the Instant Pot or an Iris Osama yogurt machine

Have a question?  Ask me.  mimi @ bravocoop dot com.

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What will Bravo do for us?

Q What does Bravo do for us and why would we want to use Bravo Yogurt?



Written by Mimi Castellanos

Bravo yogurt is made from very high-end probiotics: some call them “royal strains”.  They are arranged into a formula with yeasts and kefir and other more active ingredients that are catalysts that make it work.  Dr. Ruggiero made 324 experiments until the product ratio performed the desired results.

What happens is that viruses don’t leave when our cold or flu is over.  Sometimes we rush off too soon without a complete recovery and the viruses remain in small colonies.  These colonies slowly grow in the background and flare up when given the right environment or a big load of sugar.  (Thus our flu season is right after Christmas)

Our opponent, the virus and other bad microbes,  are small but they are very clever about their survival and existence.  They hide themselves and their nests in biofilms which are like glue.  They glue everything until they discover what works for them.  They discovered that gluing the whole Vitamin D channel is particularly helpful to them.  (Not so much for us!!)  When they glue down door 420, the immune system doesn’t come on and get after them!

Within the vitamin D channel, behind door 420 is a protein that when released, turns on the immune system!  It is called GcMAF.  When this door being glued the immune system does not turn on when we call for it.  Thus the virus and biofilm intentionally stop us from helping ourselves to be well, clean and safe. 

With microbes living inside, our consciousness may feel invaded and we may feel that you are loosing an “unknown” battle. Our consciousness is not wrong but it is often hard to understand that it really is happening somewhere.  Like in dreams, we try to assign a possible reason for this feeling.  This is like a sum total of impulse communication from many cells who are then at that moment calling out while they are at the mercy of the invading microbes.  You may come to the wrong conclusion and the war will continue unchecked if we aren’t very attentive and investigative.

Decoding these messages from the cells and responding to them is pretty easy!! Just don’t ignore it.  Do anything to help the cells and that will get you moving in the right direction!  Eventually, they will have the strength to show you their issue. Drink water, jump around.  Look at them.  Look at what you are eating.  Exercise. Release your darkness.  Do things.  Learn things.  Don’t stop until you make those cells feel better.   You need to get the disease out and you are the one who can tell where it is.  Your body can’t do it without YOU.  Gratitude will follow. 

I talk to a lot of people.  They all use the same products.  Some get well, others don’t get it.  I also use the products.  But sometimes, they don’t work as well as other times.  It isn’t my diet that makes the difference.  If I am really busy doing something else, I may slip into a state of compromised health.  When we get too busy, we get sick. I realized   I also realized that I am using my full power somewhere else and the sheer energy use will cut power from all other systems, including digestion, respiration, and the immune system.  Take care to rest, keep inside your limits and stay involved with your health.  Don’t stop this when you are well.  You will keep improving!

Bravo yogurt creates a microbiome which is easily damaged by antibiotics, prescriptions, acidic food, or stress. We need the microbiome.  It makes functional molecules for our metabolism and processing.  It helps us be connected to the world around us and so much more.  It’s like a good, protective, and nurturing parent for our body.

There isn’t any GcMAF in the jar you are taking, but it does show up when it gets to joining with the mucosa in the gut and making a microbiome.  There actually is supposed to be more beneficial microbes than anything else in our body.  These microbes come in with good quality food like the veggies with white powder on it.  That’s the probiotics.  Originally. it is built from breastfeeding where much of the immunity comes in installments and matures with the baby.  Without a microbiome, the body is defenseless.  Antibiotics can single-handedly clean out a microbiome in just 5 days!  It takes months and months of targeted probiotic therapy to restore the body’s microbiome.

The effects of not having a microbiome from taking antibiotics are important. It will leave a change in the overall feel of your body.  Antibiotics will leave you feeling empty and powerless.   After all, you lost your partner.  The way you can tell is the same way you feel yourself to know you are sick or well.  If you have a sickness, you can feel it.  Flu will leave you feeling wonky and distorted but without rest, TLC and chicken soup, you may get better but you may not notice that 10%  wonky feeling that’s left over and still growing inside.  When you take Bravo, the leftover virus (called the viral load) leaves.  This makes you much less able to get sick from colds because you don’t have a viral layer growing inside.  Removing the virus also removes the 10% wonky feeling.  If you give Bravo the time to remove all the viruses, you just feel better and better!

For me, after the viruses were gone, it began insulating my nerves and connective tissue.  It might be a lesser function for the priority of the body, but for me, I could think better and more clearly.  I could analyze a problem from 4 different directions at once again.  Then I realized, oh my, I am down to 1 angle of analysis.  Oh my, the greater realization is that I didn’t even notice my abilities were less.  UNTIL BRAVO BROUGHT THEM BACK. Again, if I had quit before the body was done, I would not have had those wonderful results!

Eat yogurt and take probiotics can’t do this by itself.  Dr. Ruggiero did 325 experiments to get Bravo Yogurt to create a microbiome that specifically makes GcMAF.  The microbiome sets up in the mucosa and starts to turn on the immune system in one area at a time.  This is another reason to go slowly.  (How many pitching machines do you want to respond to at once?).  Due to the fact that the immune system is something that is in the whole body and it is turned on and recovered a bit at a time, you will feel more and more alive as it moves through these systems.  Once it does, it actually mounts more complex issues like a connection to the global microbiome.  Stay with it.  It’s awesome.

Bravo Yogurt goes into our upper bodies down to the stomach acid.  That would be our mouths, head, esophagus, diaphragm, hearts, lungs and spines.  That’s a lot to cover!  Enteric coated capsules work far beyond the stomach and land in the solar plexus region.   Suppositories also help the lower colon, bladder, and reproduction.  The pelvis feels the echos of viruses 6 generations back and is quite uplifted by the Bravo suppositories!!!  I was a hard sell on using suppositories.  Unconsciously, I didn’t want to disturb that area. Now I realize how thick the body is at the hips and pelvis and how hard it is for the body to regulate immunity there.  I became really happy when I started using suppositories. Taking yogurt doesn’t do it.

Bravo also goes into the muscles and travels through them as it grows our colonies.  It can really help long term colonies that weaken muscles over time.  Rotator cuff melts when heat and rage are present.  E coli causes stomach ulcers similarly.  The immune system just needs some TLC, and long term dedication from you to do its job.  Don’t be a creative type that flits away.  You will feel abandoned and deeply uncared for.  That feeling of being cared for comes from YOU CARING FOR YOURSELF.

Bravo comes through.  It eventually has time to clean out all the systems including the Vitamin D channel and gets your immune system back on naturally.  Keep taking Bravo because there are always areas of toxins that need to be removed.  We don’t live in a vacuum.  Our house gets dirty, the grass in the lawn grows and weeds are inevitable.  Good quality growth is reasonable and must be monitored.

Q:  What do we have to do alongside taking Bravo?
A:  Dr. Ruggiero calls for a blood cleaner called Rubia cordifolia, aka,  Bravo Protocol which we made available for you.  Along with that, green leafy vegetables, vitamins & minerals, Vit D2&K2, and I like DHA with it too.  And lots of water so your body can transport things.   The body’s immune system loves plants and proteins.  Not so much the carbs which burn hot and dirty.  The doctor recommends a ketogenic diet which is much like paleo. 


Q: Is is important that we don’t eat a lot of yogurt or fermented things that could change this formula?

A: Yes.  It is possible to “flood” the formula with strains that change the ratio of the product’s formula.  You could bend it far enough that it won’t make GcMAF!  If you are used to something, take them at different times of the day and limit the amount if you do the “flood” method.

Q: There are people on the media that are publishing formulas to make GcMAF.  Do they work?
A:  I looked at this in 2015 (when I first looked at the product) because I always try to do a better job for less money. I personally couldn’t do that with this product, because it has not only a proprietary formula which is extensive but each ingredient is expensive and would have to be purchased by themselves.  Further, the doctor definitely had his intention of loving and healing locked into the product.  I found that the pricing was actually reasonable and much more direct than trying to fudge it.   I found myself just wanting to stand beside him and support him!!  In my heart that was the best option.   I decided, I never wanted to be without a microbiome and this is clearly the best way to do it and my machinations were unnecessary and uneventful.  The product has intention, strength, and power that imitations would never have.

Q What does this microbiome actually do?

A. The microbiome grows into the tissue and fills it with what it needs.  It brings oils, insulation, food, cleaning power, better and smoother communication, focus and makes things warm and friendly.  These are things that can’t be made by the body alone.  As it grows into an area, it brings with it what the body needs like synthesizing supplies into new tools for the immune system in each individual area it moves into.

Q:  Did the inventor Dr. Marco Ruggiero have inspiration when inventing the formula?
A:  Yes.  There was a study in Tunisia where they had people with HIV positive blood tests.  They gave them a specifically built yogurt and they got better.  (Better as in blood test with no presence of HIV positive.)  This was an area in Tunisia where food was difficult to find and yogurt was difficult to produce so they did a second experiment as the starting powder.  It didn’t work.  (The process does involve some interaction with lactoferrin.)  The capsules we sell and make are literally yogurt which is dried and put in an enteric capsule, not the starting powder. It takes a day to reconstitute in our bodies and then feels all glowing and bright.

Q: How did you know the microbiome and immune system were killing viruses?

A: For about 6 months (after being on it about as long) I felt flush like 10% of a flu every time I sat down.  I didn’t feel it all the time, or in the same place and not when I was busy.  This gave me another big clue.  The immune system needs time when I am resting to work.  When I am too busy, it waits, waits, and waits.  This isn’t how it works.  I go back to the baby breastfeeding and napping.  The perfect combination!

There is a primer about Bravo on our website in the FAQ section

How to Make it, How to take it, what to order when you first start out.

We also have a practitioner’s corner.



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What’s under the hood?  Dairy Bravo Yogurt – Ingredients

Dairy Bravo Yogurt – Ingredient Panel

Ingredient panel on the product but a look under the microscope shows at least 40 strains.

There are three packets of powder in each set which we call a 1 pack.  It makes 4 cups/1 quart/1 liter of product.

The first month is started very slowly to condition the detox pathways.  We use small amounts, therefore, only 2 cups are needed per person.  To make 2 cups, follow the same recipe but split the milk and powders in half and put the unused powder in a ziplock in the fridge for next time.  You can freeze the right amount the milk to defrost when needed.

This is the current packaging.  There are older packaging concepts and recipes on the web which may not apply.

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Prior to May 2019 there are reports of 40 different strains in the product.  This claim is not  backed up anywhere, but it is published.  Now it’s clear!! We got a bigger microscope!!

May 2019 –  There are more than 300 organisms!  Profound words by Dr. Marco Ruggiero sent to Bravo Coop as an exclusive.

In the process of carrying advanced research on our products, we conducted an extremely detailed analysis of the microbial composition of Bravo using a novel tool that exploits the most advanced concepts of molecular genetics.  We used the Axiom Microbiome array that is a chip covered in short DNA sequences that are specific to certain target organisms (a total of approximately 12,000 species are included). In short, this assay can detect up to 12,000 different microbes with extremely high specificity and sensitivity.

After having performed the current genetic analysis, we found out that there are not only approximately 40 strains, as we had been repeating for all these years. There are more than 300 (three hundred) different microbes, the highest number ever recorded in a probiotic.  (The discrepancy, in our favor, derives from the fact that under a microscope, as we had to do in the past, we could not distinguish the sub-strains, the plasmids and the phages (see later) that only the current genetic analysis can differentiate between these.)

Just to give you an example, imagine that you want to catalog the animals in a zoo. A superficial analysis (analogous to what we did with a microscope on Bravo) will tell you that there are monkeys, dogs, horses and so on. But a detailed analysis (analogous to what we did with the Axiom Microbiome array) will tell you that among monkeys you have gorillas, chimpanzees, orangutans and many other types of monkeys; among dogs, Chihuahuas, Great Danes, German shepherds and many others; among horses, Arabian pure-breeds, Mustangs, ponies, English heavy horses etc. At this point, you will be able to say that in such a zoo there is a great variety of animals; that zoo is much richer than a zoo that only has gorillas among monkeys, Chihuahuas among dogs (even though usually there are no dogs in zoos, but this is fictional), Mustangs among horses.

The Axiom Microbiome array was performed by the most reputable company in the field, Eurofins, and we believe it has legal value as far as claims are concerned since this type of analysis is also used to rule out the presence of pathogens. In other words, if we claim that Bravo is unique because it has more than 300 different microbes, and someone asks for proof, we can back this claim with the Eurofins’s analysis.

I am sure you realize that these results put Bravo in a completely different league, or better, in no league at all since no other probiotic can honestly make such a claim.

This is because Bravo, at variance with all other probiotics in the market, was designed since its inception as a means to reproduce the complexity of the healthy human microbiota.

However, it is not the sheer number of different microbes (more than 300) that is utterly impressive, but rather the biodiversity that makes Bravo even more unique. The genetic analysis revealed that Bravo, in addition to probiotic microbes, contains a great number of plasmids and phages.

Plasmids are self-replicating small DNA molecules that carry genes that benefit the survival of the organism and can be transferred to other living beings. Bravo not only contains a very high number of live probiotic microbes; thanks to the presence of plasmids, the probiotic microbes in Bravo are also able to communicate with each other and with the cells of the human body with mutual reciprocal benefits.

Thanks to the presence of plasmids, we can say that Bravo not only contains an incredibly high number of live probiotic microbes but also that these microbes are able to talk to each other and to our human cells. No other probiotic can make this claim.

However, even more, interesting could be, for its ramifications, the presence of phages.

Phages are the microbes that colonize the microbes; they can also be defined as “the microbiota of the microbiota”. The study of phages and how they relate to human health represents one of the frontiers of molecular medicine.

Phages are the most abundant organisms in the biosphere and have been of interest to scientists as tools to understand fundamental molecular biology, as vectors of horizontal gene transfer and drivers of evolution, as sources of diagnostic and genetic tools and as novel therapeutic agents.

Phages impact immunity directly, in ways that are typically anti-inflammatory. Phages modulate innate immunity via phagocytosis and cytokine responses but also impact adaptive immunity via effects on antibody production and effector polarization. Phages may thereby have profound effects on the fighting of bacterial infections or cancer growth by modulating the immune response.

Thanks to the presence of phages, we can now explain all the almost incredible clinical results that you and we have seen with Bravo.

In Bravo, probiotic microbes live together with plasmids and phages in a complex network of communication, a symbiosis that is reminiscent of the origin of life on Earth.

As you may know, we live in a microbial driven world that only exists because bacteria and Archaea tempered the previously hostile environment on early Earth to create atmospheric conditions that allow eukaryotic life forms such as humans to flourish; it has now become evident that phages were, and still are, responsible for wellbeing of microbes, which themselves impact environments at large.

In other words, if it were not for the phages – the microbes of the microbes – life would not exist, and this is why they are so important for our well being and the well being of the environment.

I hope that these notes of mine are helpful for your efforts to further highlight the unique properties of Bravo. Dr. Pacini and I are very grateful for your continuing support and we hope that having demonstrable proof of the uniqueness of Bravo, will further strengthen the success of the product.

Buy a Bravo 3 liter pack here for $95 and save!

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