**1. Introduction**

448 New Advances in the Basic and Clinical Gastroenterology

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It has been documented that the intestinal tract is inhabited by more than 1012 bacterial cells per gram of dry matter (Hayashi et al., 2002a; Langendijk et al., 1995; Suau et al., 1999), which is comprised of an estimated 400 to 500 bacterial species (Moor & Holdeman, 1974). The composition and activities of the indigenous intestinal microbiota are of paramount importance in human immunity, nutrition, and pathological processes, and therefore, the health of the individual (Van der Waaij et al., 1971). It is well established that the intestine is an important site of local immunity, and recent reports have suggested that it is a major site of extrathymic T cell differentiation (Cerf-Bensussan et al., 1985; Guy-Grand et al., 1991; Iiai eta al., 2002; Uchiyama-Tanaka, 2009). Numerous activated and quiescent lymphocytes are produced within gut-associated lymphatic tissues (GALT), such as Peyer's patches (Takahashi et al., 2005). Thus, it has been speculated that people who suffer from constipation and who harbor fecal residues in the intestine may have decreased local immune system function.

Colonic irrigations referred to as a colonics are a type of colonic hydrotherapy performed using an instrument in combination with abdominal massage, but without drugs or mechanical pressure. I previously reported that colonic irrigation may induce lymphocyte transmigration from GALT into the circulation, which may improve the functions of both the colon and immune system (Uchiyama-Tanaka, 2009). Colonic irrigation was developed about 40 years ago and no serious complications associated with its use have been reported. However, the impact of this method, which use a large amount of water, on the intestinal microbiota and serum electrolytes remains unknown. In this study, colonic irrigations were performed 3 times for each of the 10 subjects with no history of malignant or inflammatory disease.

#### **2. Materials and methods**

#### **2.1 Study design and subjects**

The procedures used in this study were in accordance with the guidelines of the Declaration of Helsinki for Human Experimentation, 2000 and all subjects provided informed consent. Ten outpatients from the Yoko Clinic (4 men and 6 women; mean age=38 ± 6 years; age range: 27–47 years) admitted to the hospital between April and May 2009 were enrolled in this study. None of the subjects had cancer or any active inflammatory disease.

The Influence of Colonic Irrigation on Human Intestinal Microbiota 451
