Abstract:
Although the inventors previously found that the novel species designated  Lactobacillus clearans  was highly effective for health purposes, they noted in particular that this species lacked sufficient intestinal purification action. Efforts to produce a species of higher potency and the like to remedy this drawback led to the unavoidable conclusion that the use of a bacterium by itself produced limited results. The concurrent use of another active substance was thus considered in seeking a lactic acid bacteria preparation capable of invigorating healthy individuals, and of renewing a sense of well being and restored health in semi-sick individuals or semi-healthy individuals. 
     The invention relates to a lactic acid bacteria preparation, comprising viable cells of  Lactobacillus clearans , and either or both of viable and killed cells of  Enterococcus faecalis  capable of reducing one or more of at least triglycerides and cholesterol.

Description:
BACKGROUND OF THE INVENTION 
     1. Field of the Invention 
     The present invention relates to a lactic acid bacteria preparation that is extremely significant for the health of humans and animals, wherein the concurrent use of  Lactobacillus clearans , which was first isolated by the inventors and belongs to the Lactobacillus genus yet has specific characteristics not found in conventionally known bacteria, and either or both of viable or killed cells of  Enterococcus faecalis  (Enterococcus), which are profoundly involved in lipid metabolism, allows the advantages of each to be more clearly and effectively brought out. 
     2. Description of the Related Art 
     There are reportedly 300 species and 100 trillion individual bacteria in the intestines, weighing a total of 1 kg and far outnumbering the total of 60 trillion human cells. Years of immunological study and research have gradually elucidated their significant involvement in the health and various diseases of humans and animals, and experts now regard intestinal flora as a vital organ function. Beneficial bacteria belonging to the group of lactic acid bacteria, such as the Bifidobacterium genus and the Lactobacillus genus, co-exist in a balanced manner when predominant. This balance can be broken down by a variety of factors, such as changes in the amount and quality of daily diet, overexertion, sleeplessness, and mental stress, resulting in the proliferation of harmful bacteria such as Welch bacillus and Veillonella. The expulsion of beneficial bacteria results in greater production of harmful substances, which can lead to a variety of illnesses and accelerated aging overall, from feces disorders such as diarrhea and constipation, to those resulting from poisoned blood, such as chronic fatigue, chapped skin, hepatic dysfunction, hypertension and arteriosclerosis. 
     The Russian Metchnikoff (1845 to 1916) theorized that the primary cause of aging was poisoning from toxins formed by intestinal putrefying fermentation, and advocated as a remedy the habit of drinking lactic acid bacteria beverages such as yogurt to help prevent aging. Throughout the vagaries of the history of lactic acid bacteria since then, it has not been possible to grasp the true practical utility of such bacteria. That is because matters now revealed by immunological studies and elucidated by experiments could not be unraveled. 
     Metchnikoff&#39;s hypothesis has been borne out and is now common knowledge in the health sciences, as recent progress in microbiology has elucidated a variety of important functions by beneficial intestinal bacteria, such as the reduction in intestinal acidity and the promotion of intestinal motility to help in the digestion of foods and the absorption of nutrients, the simultaneous suppression and degradation of harmful substances, the synthesis of vitamins and amino acids, protection against intestinal infections by pathogens, and enhanced immunological power. 
     In addition, along with recent health trends, lactic acid bacteria have established a firm footing as beverages and antiflatulents for many people. It cannot be denied, however, that such products do not appeal to people who are looking for positive results, having failed to realize better health and improved symptoms through daily ingestion or use of lactic acid bacteria. Unfortunately, lactic acid bacteria products are still consumed merely as refreshment, or still tend to be regarded as a preference similar to coffee. 
     In view of the foregoing, there has been a need for a product which could be rapidly administered to bring out better effects unavailable in conventional lactic acid bacterial beverages or preparations. Although the inventors previously found that the novel species designated  Lactobacillus clearans  was highly effective for health purposes, they noted in particular that this species lacked sufficient intestinal purification action. Efforts to produce a strain of higher potency and the like to remedy this drawback led to the unavoidable conclusion that the use of the bacterium by itself produced limited results. The concurrent use of another active substance was thus considered in seeking a lactic acid bacteria preparation capable of invigorating healthy individuals, and of renewing a sense of well being and restored health in semi-sick individuals or semi-healthy individuals. 
     SUMMARY OF THE INVENTION 
     As a result of extensive research to find a compatible biological or non-biological partner to remedy the aforementioned drawbacks, the inventors perfected the present invention upon finding, among  Enterococcus faecalis  belonging to the same group of lactobacilli as  Lactobacillus clearans , a species capable of reducing at least one among triglycerides and cholesterol. That is, the present invention is a lactic acid bacteria preparation, comprising viable cells of  Lactobacillus clearans , and viable cells of  Enterococcus faecalis  capable of reducing one or more of at least triglycerides and cholesterol. 
     The second of the inventions is a lactic acid bacteria preparation, comprising viable cells of  Lactobacillus clearans , and killed cells of  Enterococcus faecalis  capable of reducing one or more of at least triglycerides and cholesterol. 
     The third of the inventions is a lactic acid bacteria preparation, comprising viable cells of  Lactobacillus clearans , and viable and killed cells of  Enterococcus faecalis  capable of reducing one or more of at least triglycerides and cholesterol. 
     DESCRIPTION OF THE PREFERRED EMBODIMENTS 
     The  Lactobacillus clearans  referred to in the present invention is a novel strain belonging to the Lactobacillus genus, and has the following biochemical characteristics 1, 2, 3, and 4. Specifically, it comprises strains of the Lactobacillus genus that 1) are capable of reducing both Na 2 S.9H 2 O and NH 4 OH when 0.5 g Na 2 S.9H 2 O and/or 0.5 mL NH 4 OH is or are added to 5 g meat extract, 5 g peptone, 1 g glucose, 1 g CaCO 3 , and 1 L water (neutral pH); 2) show no growth-promoting action despite the addition of 0.5 g Na 2 S.9H 2 O and/or 0.5 mL NH 4 OH during the logarithmic growth phase of bacterial culture in medium comprising 1 g casamino acid and vitamins (A: 900 IU; B 1 : 1 mg; B 2 : 1 mg; B 6 : 1 mg; B 12 : 5 γ; nicotinamide: 16 mg; calcium pantothenate: 8 mg; C: 64 mg; and D 2 : 120 IU) in Stephenson-Wetham medium ((abbreviated as S-W) 1 g KH 2 PO 4 , 0.7 g MgSO 4 .7H 2 O, 1 g NaCl, 4 g (NH 4 ) 2 HPO 4 , 0.03 FeSO 4 .7H 2 O, and 5 g glucose); 3) have resistance, in the form of naturally isolated strains, to Na 2 S.9H 2 O greater than that of conventionally known lactic acid bacteria but weaker than that of  Lactobacillus deodorans ; and 4) are gram positive, rod-shaped, non-motile, and catalase negative, are not nitrate-reducing and not gelatin-degrading, do not produce indole or hydrogen sulfide, have a high capacity for forming lactic acid from glucose and lactose, and have growth promoted by the addition of acetic acid (see Japanese Patent Publication(Kokoku) H4-632. 
     Advantages of  Lactobacillus clearans , in addition to the action of common Lactobacillus, include potent intestinal purification action absent in conventional Lactobacillus as described in Japanese Patent 1714413 and Japanese Patent Application 11-15177, such as: 1) decomposition of intestinal putrefying malodorous substances; 2) increases in beneficial bacteria such as Bifidobacterium and Lactobacillus among the intestinal flora, with dramatic reductions in harmful bacteria such as Welch bacillus and Veillonella, thereby providing a better balance in the intestinal flora to improve the intestinal environment; and 3) suppression of the growth primarily of etiologic agents causing intestinal infections, and diminished toxicity. 
     This action will be described below through tests. In vitro tests were conducted first. 15 mL of a 10-fold dilution of feces was introduced into test tubes (18×180 mm). The test tubes were then inoculated with sample bacteria 1) that had been sterilized at high pressure for 15 minutes at 120° C. and 2) that had not been sterilized. The test tubes were stoppered with rubber plugs for 72 hours of anaerobic culture at 37° C. 1 mL air was then drawn by syringe from inside the test tubes and injected into 5 L odor bags for olfactory tests by a panel of 6 individuals. The odor was assessed based on the feces odor criteria given in Table 1. 
     
       
         
               
             
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Feces odor criteria 
               
             
          
           
               
                 Odor 
                   
               
               
                 ranks 
                 Odor 
               
               
                   
               
               
                 1 
                 faint odor recognizable as that of feces 
               
               
                 2 
                 easily recognizable as slight odor of feces 
               
               
                 3 
                 clearly recognizable as weak odor of feces 
               
               
                 4 
                 strong odor equal to untreated feces 
               
               
                 5 
                 extremely strong odor more pronounced than untreated feces 
               
               
                   
               
             
          
         
       
     
     Three typical strains of  Lactobacillus clearans , specifically, FERM BP-6972, FERM BP-6971, FERM BP-6973, were used to test the deodorizing capacity in the aforementioned in vitro tests. The results in Table 2 show that the samples sterilized at high pressure were clearly deodorized, with an odor assessed as being the weak or slight odor of feces. The unsterilized samples, on the other hand, showed less deodorization than the samples sterilized at high pressure, because malodorous substances were produced, perhaps as a result of the activity of numerous putrefying bacteria in the feces, but they too were nevertheless clearly deodorized. The results for the 3 typical strains of  Lactobacillus clearans  in the aforementioned test were similarly reproduced with other similar strains. 
     
       
         
               
             
               
               
               
             
               
               
               
             
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 Deodorizing capacity of  Lactobacillus   
               
               
                   clearans  (in vitro) 
               
             
          
           
               
                 Inoculum strain 
                 Odor (average value by panel) 
                   
               
             
          
           
               
                 FERM BP-No. 
                 High pressure sterilization 
                 Unsterilized 
               
               
                   
               
             
          
           
               
                 6972 
                 2.5 
                 2.8 
               
               
                 6971 
                 2.3 
                 2.5 
               
               
                 6973 
                 2.0 
                 2.3 
               
               
                 Not inoculated 
                 4.0 
                 4.5 
               
               
                   
               
             
          
         
       
     
     In subsequent in vivo tests, 150 mL yogurt prepared using  Lactobacillus clearans  FERM BP-6973 was ingested once a day, and the odor of feces was determined 20 to 30 days, 50 to 60 days, and 80 to 90 days after ingestion, with the averages given in Table 3. The results were determined by introducing 0.5 g feces samples from the 10 individuals into 20 L odor bags, which were stored at room temperature for evaluation of the odor by a panel of 6 individuals after 30 minutes. The average feces odor for 5 individuals ingesting no yogurt was rated as 100. Although feces odor varied considerably depending on the dietary contents, the average odor for 10 individuals ingesting yogurt decreased about 50% 1 month after ingestion, 70% after 2 months, and 85% after 3 months. After 3 months, however, continuous ingestion resulted in a peak odor reduction of 80 to 90%. 
     
       
         
               
             
               
               
             
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 Feces odor after ingestion of yogurt prepared 
               
               
                 using  Lactobacillus clearans   
               
             
          
           
               
                 Odor when no yogurt 
                 Odor after ingestion of yogurt 
               
             
          
           
               
                 ingested 
                 20 to 30 days 
                 50 to 60 days 
                 80 to 90 days 
               
               
                   
               
               
                 100 
                 50 
                 30 
                 15 
               
               
                   
               
             
          
         
       
     
     In terms of the correlation between purification and bacteria in the natural world, the inventors narrowed the analyzed substances into easily testable malodorous substances, which were broadly classified into sulfur compounds, nitrogen compounds, and carbon compounds. It was found that bacteria capable of degrading malodorous sulfur compounds such as Na 2 S.9H 2 O, malodorous nitrogen compounds such as NH 3 , and malodorous carbon compounds such as acetic acid, butyric acid, and similar lower fatty acids were capable of degrading most malodorous sulfur, nitrogen, and carbon compounds of polymers therefrom, this being referred to as the SNC theory. The following tests were conducted in accordance with this theory. 
     0.5 g Na 2 S.9H 2 O or 0.5 mL ammonia water (NH 4 OH) was added to 1 mL synthetic medium comprising 5 g meat extract, 5 g peptone, 3 g sodium butyrate, 5 g glucose, and 3 g CaCO 3 , the medium was inoculated with  Lactobacillus clearans  for 72 hours of anaerobic culture at 37° C., and the decrease in the Na 2 S.9H 2 O or NH 4 OH that had been added was determined over time. The Na 2 S.9H 2 O was measured by the iodine titration method of JIS K0102-1985, and the NH 4 OH was measured by the indole phenol blue absorbance method of JIS K0102-1985. The results are given in Tables 4 and 5. The tables show that  Lactobacillus clearans  had the capacity to reduce toxic malodorous Na 2 S.9H 2 O and NH 4 OH 40 to 50% in 72 hours. This means that it can degrade and assimilate most other malodorous toxic substances. 
     
       
         
               
             
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 4 
               
             
             
               
                   
               
               
                 Degradation and assimilation of sodium disulfide 
               
               
                 by  Lactobacillus clearans   
               
             
          
           
               
                 Inoculum 
                 Concen- 
                   
               
               
                 strain 
                 tration 
                 Na 2 S.9H 2 O concentration 
               
               
                 FERM BP- 
                 when 
                 and percent decrease 
               
             
          
           
               
                 No. 
                 added 
                 24 hours 
                 48 hours 
                 72 hours 
               
               
                   
               
               
                 6972 
                 500 
                 400 ppm 
                 350 ppm 
                 300 ppm 
               
               
                   
                 ppm 
                 20% decrease 
                 30% decrease 
                 40% decrease 
               
               
                 6971 
                 500 
                 380 ppm 
                 330 ppm 
                 275 ppm 
               
               
                   
                 ppm 
                 24% decrease 
                 34% decrease 
                 45% decrease 
               
               
                 6973 
                 500 
                 350 ppm 
                 300 ppm 
                 250 ppm 
               
               
                   
                 ppm 
                 30% decrease 
                 40% decrease 
                 50% decrease 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 Degradation and assimilation of ammonia by 
               
               
                 
                   Lactobacillus clearans 
                 
               
             
          
           
               
                 Inoculum 
                 Concen- 
                   
               
               
                 strain 
                 tration 
                 NH 4 OH concentration 
               
               
                 FERM BP- 
                 when 
                 and percent decrease 
               
             
          
           
               
                 No. 
                 added 
                 24 hours 
                 48 hours 
                 72 hours 
               
               
                   
               
               
                 6972 
                 500 
                 400 ppm 
                 350 ppm 
                 300 ppm 
               
               
                   
                 ppm 
                 20% decrease 
                 30% decrease 
                 40% decrease 
               
               
                 6971 
                 500 
                 375 ppm 
                 300 ppm 
                 280 ppm 
               
               
                   
                 ppm 
                 25% decrease 
                 40% decrease 
                 44% decrease 
               
               
                 6973 
                 500 
                 375 ppm 
                 300 ppm 
                 250 ppm 
               
               
                   
                 ppm 
                 25% decrease 
                 40% decrease 
                 50% decrease 
               
               
                   
               
             
          
         
       
     
     The above in vitro sensory tests and chemical analysis were described as methods for assessing the deodorizing action of  Lactobacillus clearans . The 10-fold dilutions of feces samples prepared at this time were centrifuged before culture and 72 hours after culture, 5 mL supernatant was collected and diluted 10-fold to determine the S 2+  and NH 4   +  ion concentrations, and the extent of their decrease was calculated. The results in Table 6 show that the decrease in the S 2+  and NH 4   +  ions in feces were even higher than the decrease in the aforementioned synthetic medium, indicating that feces was a more suitable habitat for  Lactobacillus clearans . The S 2+  ions were determined by the iodine titration method of JIS K0102-1985, and the NH 4   +  ions were determined by the indole phenol blue absorbance method of JIS K0102-1985. 
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                 Decrease in S 2+  and NH 4   +  by  Lactobacillus clearans   
               
             
          
           
               
                   
                   
                 S 2+   
                   
                 NH 4   +   
               
               
                   
                   
                 concentration 
                   
                 concentration 
               
               
                 10-fold 
                 Inoculum 
                 and decrease 
                   
                 and decrease 
               
             
          
           
               
                 feces 
                 FERM BP- 
                 Before 
                 72 hours 
                 Before 
                 72 hours 
               
               
                 dilution 
                 No. 
                 culture 
                 culture 
                 culture 
                 culture 
               
               
                   
               
               
                 Sterilized 
                 6972 
                 30 ppm 
                 15 ppm 
                 350 ppm 
                 190 ppm 
               
               
                 at 
                   
                   
                 50% 
                   
                 45% 
               
               
                 high 
                   
                   
                 decrease 
                   
                 decrease 
               
               
                 pressure 
                 6971 
                 30 ppm 
                 12 ppm 
                 350 ppm 
                 130 ppm 
               
               
                   
                   
                   
                 60% 
                   
                 63% 
               
               
                   
                   
                   
                 decrease 
                   
                 decrease 
               
               
                   
                 6973 
                 30 ppm 
                 10 ppm 
                 350 ppm 
                 110 ppm 
               
               
                   
                   
                   
                 67% 
                   
                 68% 
               
               
                   
                   
                   
                 decrease 
                   
                 decrease 
               
               
                 Not 
                 6972 
                 52 ppm 
                 35 ppm 
                 385 ppm 
                 235 ppm 
               
               
                 sterilized 
                   
                   
                 33% 
                   
                 40% 
               
               
                   
                   
                   
                 decrease 
                   
                 decrease 
               
               
                   
                 6971 
                 52 ppm 
                 30 ppm 
                 385 ppm 
                 200 ppm 
               
               
                   
                   
                   
                 42% 
                   
                 48% 
               
               
                   
                   
                   
                 decrease 
                   
                 decrease 
               
               
                   
                 6973 
                 52 ppm 
                 25 ppm 
                 385 ppm 
                 165 ppm 
               
               
                   
                   
                   
                 52% 
                   
                 57% 
               
               
                   
                   
                   
                 decrease 
                   
                 decrease 
               
               
                   
               
             
          
         
       
     
     2 μL of the aforementioned supernatant was introduced into a gas chromatograph to analyze the lower fatty acids. The gas chromatography involved the use of Col. unisole F-200 30/60 glass (3 §×3 m) at a carrier gas rate of 50 mL/min (He), with 152 kPa (0.55 kg/cm 2 G) hydrogen and 152 kPa (0.55 kg/cm 2 G) air, at a column temperature of 140° C. and Inj 200° C. The assayed substances were acetic acid, propionic acid, iso-butyric acid, n-butyric acid, iso-valeric acid, and n-valeric acid, with the calculated concentrations given in Table 7. Table 7 shows that the concentrations of lower fatty acids decreased 50 to 75% with samples sterilized at high pressure, and 40 to 60% with unsterilized samples. 
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 7 
               
             
             
               
                   
               
               
                 Decrease in lower fatty acids by  Lactobacillus clearans   
               
             
          
           
               
                   
                   
                   
                 Total lower fatty acid 
                   
               
               
                   
                   
                 Inoculum 
                 concentration and decrease 
               
             
          
           
               
                   
                 10-fold feces 
                 FERM 
                 Before 
                 72 hours 
               
               
                   
                 dilution 
                 BP-No. 
                 culture 
                 culture 
               
               
                   
                   
               
               
                   
                 Sterilized 
                 6972 
                 2850 ppm 
                 1500 ppm 
               
               
                   
                 at high 
                   
                   
                 47% decrease 
               
               
                   
                 pressure 
                 6971 
                 2850 ppm 
                  960 ppm 
               
               
                   
                   
                   
                   
                 66% decrease 
               
               
                   
                   
                 6973 
                 2850 ppm 
                  750 ppm 
               
               
                   
                   
                   
                   
                 74% decrease 
               
               
                   
                   
                 6972 
                 3500 ppm 
                 2100 ppm 
               
               
                   
                   
                   
                   
                 40% decrease 
               
               
                   
                 Not 
                 6971 
                 3500 ppm 
                 1800 ppm 
               
               
                   
                 sterilized 
                   
                   
                 48% decrease 
               
               
                   
                   
                 6973 
                 3500 ppm 
                 1280 ppm 
               
               
                   
                   
                   
                   
                 63% decrease 
               
               
                   
                   
               
             
          
         
       
     
     Lyophilized cells of three typical strains of  Lactobacillus clearans , that is, FERM BP-6972, 6971, and 6973, were prepared, and were mixed in equal parts to form a preparation. 2 g of the preparation (5×10 8  cells/g) was taken continuously, and the changes in the cell count (cells/1 g feces) of Bifidobacterium and Lactobacillus (except for  Lactobacillus clearans ) which are known as typical beneficial bacteria in the intestinal flora, as well as of typical harmful bacteria such as Veillonella and  Clostridium perfringens  (Welch bacillus), were measured over time. Table 8 shows the results of oral administration to 20 healthy individuals, and Table 9 shows the results of oral administration to 20 constitutionally weak individuals. 
     
       
         
               
             
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 8 
               
             
             
               
                   
               
               
                 Effects of oral administration of  actobacillus clearans  preparation on intestinal 
               
               
                 flora of healthy individuals mean values for 20 healthy individuals) 
               
             
          
           
               
                   
                 Cell count 
                   
               
               
                   
                 before 
                 Cell count after administration 
               
             
          
           
               
                   
                 administration 
                 1 month 
                 2 months 
                 3 months 
                 6 months 
                 12 months 
               
               
                   
                   
               
             
          
           
               
                 Beneficial 
                 Bifidobacterium 
                 1.2 × 10 10   
                 1.5 × 10 10   
                 1.8 × 10 10   
                 2.1 × 10 10   
                 2.4 × 10 10   
                 2.8 × 10 10   
               
               
                 bacteria 
                 Lactobacillus 
                   2 × 10 7   
                 2.5 × 10 7    
                   3 × 10 7   
                   5 × 10 7   
                   1 × 10 8   
                   2 × 10 8   
               
               
                 Harmful bacteria 
                 Clostridium 
                   1 × 10 5   
                   8 × 10 4   
                   7 × 10 4   
                   5 × 10 4   
                   2 × 10 4   
                   1 × 10 4   
               
               
                   
                 Veillonella 
                   5 × 10 5   
                 4.5 × 10 5    
                   4 × 10 5   
                   3 × 10 5   
                   2 × 10 5   
                   1 × 10 5   
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 9 
               
             
             
               
                   
               
               
                 Effects of oral administration of  Lactobacillus clearans  preparation on intestinal 
               
               
                 flora of constitutionally weak individuals (mean values for 20 constitutionally weak 
               
               
                 individuals) 
               
             
          
           
               
                   
                 Cell count 
                   
               
               
                   
                 before 
                 Cell count after administration 
               
             
          
           
               
                   
                 administration 
                 1 month 
                 2 months 
                 3 months 
                 6 months 
                 12 months 
               
               
                   
                   
               
             
          
           
               
                 Beneficial 
                 Bifidobacterium 
                 5 × 10 9   
                   5 × 10 9   
                   6 × 10 9   
                 6.5 × 10 9   
                 7.5 × 10 9   
                 1 × 10 10   
               
               
                 bacteria 
                 Lactobacillus 
                 1 × 10 7   
                 1.2 × 10 7   
                 1.5 × 10 7   
                 2.5 × 10 7   
                   3 × 10 7   
                 7 × 10 7   
               
               
                 Harmful bacteria 
                 Clostridium 
                 5 × 10 5   
                 3.5 × 10 5   
                   3 × 10 5   
                 2.5 × 10 5   
                   2 × 10 5   
                 1 × 10 5   
               
               
                   
                 Veillonella 
                 2 × 10 6   
                 1.7 × 10 6   
                 1.4 × 10 6   
                 1.1 × 10 6   
                   5 × 10 5   
                 3 × 10 5   
               
               
                   
               
             
          
         
       
     
     Tables 8 and 9 show that before the administration of the  Lactobacillus clearans  preparations, the beneficial bacteria among the intestinal flora were an average of 220% greater in healthy individuals than in constitutionally weak individuals, whereas harmful bacteria were only about 22.5%. This shows that the state of the intestinal flora is an index of the current state of health, that is, how deeply involved the intestinal flora are in health. A new finding was that ingestion of the preparation first resulted in an increase in beneficial bacterial in healthy individuals, and that with this increase, there was a gradual decrease in harmful bacteria. In contrast to this pattern, the pattern found in constitutionally weak individuals was that the harmful bacteria first decreased, followed by a gradual increase in beneficial bacteria. Overall, the average changes in beneficial and harmful bacteria reflected an increase in beneficial bacteria and a corresponding decrease in harmful bacteria. This trend accelerated after 3 months, with Bifidobacterium increasing 50% after 6 months and 110% after 1 year, and with Lactobacillus increasing 300% after 6 months and 950% after 1 year. In contrast, the Clostridium decreased 70% after 6 months and 85% after 1 year, while the Veillonella decreased 67.5% after 6 months and 82.5% after 1 year. 
       Escherichia coli  O-157 , Salmonella enteitidis , and  Shigella flexneri  were cultured alone and together with  Lactobacillus clearans  (FERM BP-6973) to check the action of  Lactobacillus clearans  against pathogens, and the results were compared. The medium composition used for the cultures comprised 10 g meat extract, 10 g peptone, 2 g glucose, 2 g NaCl, and 1 g CaCO 3  per liter, with the pH adjusted to 7.2. Anaerobic culture was performed at 37° C., with subcultures repeated every 72 hours, at which point the plates were diluted and smeared to monitor the changes in cell count, whether the colonies were the S type (original) or had mutated into the R type with diminished toxicity, their ratio, and the like. The pathogens used in the tests were acquired from Medic KK (registered sanitation research institute). 
     Table 10 shows the results of  E. coli  O-157 cultured by itself, and Table 11 shows the results of mixed culture of  E. coli  O-157 and  Lactobacillus clearans . Table 10 shows that in cultures of  E. coli  O-157 alone, the cell count was a virtually constant 4 to 5×10 9  cells/g throughout 20 subcultures, with no R types showing up. Table 11 shows that in mixed cultures of  E. coli  O-157 and  Lactobacillus clearans , there was little change in the  Lactobacillus clearans  cell count, but that there were significant changes in the cell count of the  E. coli  O-157. R types showed up at the 5 th  subculture, and the ratio of R types increased with the accumulation of subsequent subcultures, until all cells formed R types by the 18 th  subculture. There was no reversion to the S type in subsequently continued subcultures. 
     
       
         
               
             
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 10 
               
             
             
               
                   
               
               
                 Results of cultures of  E. coli  O-157 by itself 
               
             
          
           
               
                   
                   E. coli  O-157 
               
             
          
           
               
                   
                 Number of 
                 Number of 
                 Number of 
                 Ratio of 
               
               
                   
                 subcultures 
                 S type cells 
                 R type cells 
                 R type cells 
               
               
                   
                   
               
             
          
           
               
                   
                 1 
                   5 × 10 9   
                 0 
                 0% 
               
               
                   
                 3 
                 4.5 × 10 9   
                 0 
                 0% 
               
               
                   
                 5 
                 4.7 × 10 9   
                 0 
                 0% 
               
               
                   
                 7 
                 4.4 × 10 9   
                 0 
                 0% 
               
               
                   
                 10 
                 4.3 × 10 9   
                 0 
                 0% 
               
               
                   
                 12 
                 4.5 × 10 9   
                 0 
                 0% 
               
               
                   
                 15 
                   4 × 10 9   
                 0 
                 0% 
               
               
                   
                 18 
                 3.8 × 10 9   
                 0 
                 0% 
               
               
                   
                 20 
                 4.2 × 10 9   
                 0 
                 0% 
               
               
                   
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 11 
               
             
             
               
                   
               
               
                 Results of cultures of mixed cultures of  E. coli   
               
               
                 O-157 and  Lactobacillus clearans   
               
             
          
           
               
                 Number 
                 Cell count 
                   E. coli  O-157 
                   
               
             
          
           
               
                 of 
                 of FERM 
                 Number of 
                 Number of 
                 Number of 
               
               
                 subcultures 
                 BP-6973 
                 S type cells 
                 R type cells 
                 R type cells 
               
               
                   
               
             
          
           
               
                 1 
                 1.2 × 10 9   
                 5 × 10 9   
                 0 
                  0% 
               
               
                 3 
                 1.2 × 10 9   
                 3.5 × 10 9    
                 0 
                  0% 
               
               
                 5 
                 1.4 × 10 9   
                 0 
                 1.9 × 10 9   
                 100%  
               
               
                 7 
                   1 × 10 9   
                 1 × 10 9   
                   1 × 10 9   
                 50% 
               
               
                 10 
                   7 × 10 8   
                 1.2 × 10 9    
                 2.2 × 10 9   
                 65% 
               
               
                 12 
                 1.1 × 10 9   
                 5 × 10 8   
                   2 × 10 9   
                 80% 
               
               
                 15 
                 1.2 × 10 9   
                 2 × 10 8   
                 3.2 × 10 9   
                 94% 
               
               
                 18 
                 1.5 × 10 9   
                 0 
                 1.7 × 10 9   
                 100%  
               
               
                 20 
                 1.3 × 10 9   
                 0 
                 1.2 × 10 9   
                 100%  
               
               
                   
               
             
          
         
       
     
     Table 12 shows the results obtained with  Salmonella enteritidis  cultured by itself, and Table 13 shows the results obtained with mixed cultures of  Salmonella enteritidis  and  Lactobacillus clearans . Table 12 shows that in cultures of  Salmonella enteritidis  alone, the cell count was 3 to 5×10 9  cells/g. R types showed up sponeously in the 14 th  subculture, reaching a ratio of 5% by the 20 th  subculture. The ratio increased in subsequent subcultures, peaking at 23% by the 50 th  subculture. The ratio of R types thereafter stayed at about 20%. Table 13 shows that in mixed cultures of  Salmonella enteritidis  and  Lactobacillus clearans , the ratio of R types was 29% in the 5 th  subculture, and that the ratio of R types increased with the accumulation of subsequent subcultures, reaching 50% in the 10 th  subculture, 90% in the 20 th  subculture, and 100% in the 47 th  subculture. S type reversion in subsequent subcultures was less than 1%. After the 70 th  subculture, all S types disappeared, with no further reversion. 
     
       
         
               
             
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 12 
               
             
             
               
                   
               
               
                 Results of cultures of  Salmonella enteritidis  by itself 
               
             
          
           
               
                   
                 
                   Salmonella enteritidis 
                 
               
             
          
           
               
                   
                 Number of 
                 Number of 
                 Number of 
                 Ratio of 
               
               
                   
                 subcultures 
                 S type cells 
                 R type cells 
                 R type cells 
               
               
                   
                   
               
             
          
           
               
                   
                 1 
                 5 × 10 9   
                 0 
                  0% 
               
               
                   
                 5 
                 4 × 10 9   
                 0 
                  0% 
               
               
                   
                 10 
                 3 × 10 9   
                 0 
                  0% 
               
               
                   
                 15 
                 3 × 10 9   
                 1 × 10 8   
                  3% 
               
               
                   
                 20 
                 3 × 10 9   
                 1.5 × 10 8    
                  5% 
               
               
                   
                 25 
                 3.5 × 10 9    
                 2 × 10 8   
                  5% 
               
               
                   
                 30 
                 4 × 10 9   
                 5 × 10 7   
                  1% 
               
               
                   
                 35 
                 3.7 × 10 9    
                 3 × 10 8   
                  8% 
               
               
                   
                 40 
                 3.2 × 10 9    
                 5 × 10 8   
                 14% 
               
               
                   
                 47 
                 3 × 10 9   
                 7 × 10 8   
                 19% 
               
               
                   
                 50 
                 3.3 × 10 9    
                 1 × 10 9   
                 23% 
               
               
                   
                 55 
                 2.8 × 10 9    
                 8 × 10 8   
                 22% 
               
               
                   
                 60 
                 3 × 10 9   
                 7 × 10 8   
                 17% 
               
               
                   
                 65 
                 3.2 × 10 9    
                 5 × 10 8   
                 14% 
               
               
                   
                 70 
                 2.8 × 10 9    
                 6 × 10 8   
                 18% 
               
               
                   
                 75 
                 3 × 10 9   
                 7 × 10 8   
                 19% 
               
               
                   
                 80 
                 2.7 × 10 9    
                 5 × 10 8   
                 16% 
               
               
                   
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 13 
               
             
             
               
                   
               
               
                 Results of mixed cultures of  Salmonella enteritidis   
               
               
                 and  Lactobacillus clearans   
               
             
          
           
               
                   
                 Cell count 
                 
                   Salmonella enteritidis 
                 
               
             
          
           
               
                 Number of 
                 of FERM 
                 Number of 
                 Number of 
                 Ratio of 
               
               
                 Subcultures 
                 BP-6973 
                 S type cells 
                 R type cells 
                 R type cells 
               
               
                   
               
             
          
           
               
                 1 
                 1 × 10 9   
                 5 × 10 9   
                 0 
                  0% 
               
               
                 5 
                 5 × 10 8   
                 2.5 × 10 9    
                 1 × 10 9   
                 29% 
               
               
                 10 
                 3 × 10 8   
                 1 × 10 9   
                 1 × 10 9   
                 50% 
               
               
                 15 
                 4 × 10 8   
                 5 × 10 8   
                 1.5 × 10 9    
                 75% 
               
               
                 20 
                 3 × 10 8   
                 2 × 10 8   
                 2 × 10 9   
                 90% 
               
               
                 25 
                 3.5 × 10 8    
                 1.5 × 10 8    
                 2 × 10 9   
                 93% 
               
               
                 30 
                 3 × 10 8   
                 1.6 × 10 7    
                 8 × 10 9   
                 98% 
               
               
                 35 
                 2 × 10 8   
                 1.5 × 10 8    
                 1 × 10 9   
                 87% 
               
               
                 40 
                 2.5 × 10 8    
                 1 × 10 8   
                 1.5 × 10 9    
                 94% 
               
               
                 47 
                 3 × 10 8   
                 0 
                 2 × 10 9   
                 100%  
               
               
                 50 
                 5 × 10 8   
                 3 × 10 7   
                 2 × 10 9   
                 99% 
               
               
                 55 
                 6 × 10 8   
                 0 
                 2.1 × 10 9    
                 100%  
               
               
                 60 
                 5 × 10 8   
                 1 × 10 7   
                 1.8 × 10 9    
                 99% 
               
               
                 65 
                 7.5 × 10 8    
                 2 × 10 7   
                 2 × 10 9   
                 99% 
               
               
                 70 
                 8 × 10 8   
                 0 
                 2.2 × 10 9    
                 100%  
               
               
                 75 
                 7 × 10 8   
                 0 
                 2 × 10 9   
                 100%  
               
               
                 80 
                 7.5 × 10 8    
                 0 
                 1.8 × 10 9    
                 100%  
               
               
                   
               
             
          
         
       
     
     Table 14 shows the results of  Shigella flexneri  cultured by itself, and Table 15 shows the results of mixed culture of  Shigella flexneri  and  Lactobacillus clearans . Table 14 shows that in cultures of  Shigella flexneri  alone, R types showed up spontaneously in the 10 th  subculture, reaching a ratio of 9% by the 20 th  subculture. The ratio increased in subsequent subcultures, peaking at 20% by the 40 th  subculture. The ratio of R types thereafter stayed at about 10 to 20%. Table 15 shows that in mixed cultures of  Shigella flexneri  and  Lactobacillus clearans , R types showed up in the 5 th  subculture, and that the ratio of R types increased with the accumulation of subsequent subcultures, reaching 100% in the 80 th  subculture. There was S type reversion in subsequent subcultures, but less than 1%. After the 108 th  subculture, all S types disappeared, with no further reversion. 
     
       
         
               
             
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 14 
               
             
             
               
                   
               
               
                 Results of cultures of  Shigella flexneri  by itself 
               
             
          
           
               
                   
                 
                   Shigella flexneri 
                 
               
             
          
           
               
                   
                 Number of 
                 Number of 
                 Number of 
                 Ratio of 
               
               
                   
                 Subcultures 
                 S type cells 
                 R type cells 
                 R type cells 
               
               
                   
                   
               
             
          
           
               
                   
                 1 
                   5 × 10 9   
                 0 
                  0% 
               
               
                   
                 5 
                   5 × 10 9   
                 0 
                  0% 
               
               
                   
                 10 
                   4 × 10 9   
                 5 × 10 7   
                  1% 
               
               
                   
                 15 
                   4 × 10 9   
                 1 × 10 8   
                  2% 
               
               
                   
                 20 
                 4.2 × 10 9   
                 4 × 10 8   
                  9% 
               
               
                   
                 25 
                 3.8 × 10 9   
                 5 × 10 8   
                 12% 
               
               
                   
                 30 
                 3.7 × 10 9   
                 6 × 10 8   
                 14% 
               
               
                   
                 35 
                 3.8 × 10 9   
                 5 × 10 8   
                 12% 
               
               
                   
                 40 
                   4 × 10 9   
                 1 × 10 9   
                 20% 
               
               
                   
                 45 
                   4 × 10 9   
                 7 × 10 8   
                 15% 
               
               
                   
                 50 
                 4.2 × 10 9   
                 7 × 10 8   
                 14% 
               
               
                   
                 55 
                   4 × 10 9   
                 1 × 10 8   
                 20% 
               
               
                   
                 75 
                 3.6 × 10 9   
                 8 × 10 8   
                 18% 
               
               
                   
                 80 
                 3.5 × 10 9   
                 7 × 10 8   
                 17% 
               
               
                   
                 90 
                 3.5 × 10 9   
                 6 × 10 8   
                 15% 
               
               
                   
                 100 
                 3.2 × 10 9   
                 6 × 10 8   
                 16% 
               
               
                   
                 110 
                 3.7 × 10 9   
                 7 × 10 8   
                 16% 
               
               
                   
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 15 
               
             
             
               
                   
               
               
                 Results of mixed cultures of  Shigella flexneri   
               
               
                 and  Lactobacillus clearans   
               
             
          
           
               
                   
                 Cell count 
                 
                   Shigella flexneri 
                 
               
             
          
           
               
                 Number of 
                 of FERM 
                 Number of 
                 Number of 
                 Ratio of 
               
               
                 Subcultures 
                 BP-6973 
                 S type cells 
                 R type cells 
                 R type cells 
               
               
                   
               
             
          
           
               
                 1 
                 1.2 × 10 9   
                 5 × 10 9   
                 0 
                  0% 
               
               
                 5 
                   8 × 10 8   
                 3 × 10 9   
                 2 × 10 8   
                  6% 
               
               
                 10 
                   5 × 10 8   
                 2.5 × 10 9    
                 1 × 10 9   
                 29% 
               
               
                 15 
                   3 × 10 8   
                 2.5 × 10 9    
                 1.5 × 10 9    
                 38% 
               
               
                 20 
                   3 × 10 8   
                 2 × 10 9   
                 2 × 10 9   
                 50% 
               
               
                 25 
                   3 × 10 8   
                 1.8 × 10 9    
                 2 × 10 9   
                 53% 
               
               
                 30 
                 3.5 × 10 8   
                 1.8 × 10 9    
                 2.2 × 10 9    
                 55% 
               
               
                 35 
                   3 × 10 8   
                 1 × 10 9   
                 2.5 × 10 9    
                 71% 
               
               
                 40 
                 2.8 × 10 8   
                 8 × 10 8   
                 3 × 10 9   
                 79% 
               
               
                 45 
                 2.6 × 10 8   
                 5 × 10 8   
                 3 × 10 9   
                 85% 
               
               
                 50 
                   3 × 10 8   
                 1 × 10 9   
                 2.8 × 10 9    
                 74% 
               
               
                 55 
                 3.2 × 10 8   
                 5 × 10 8   
                 3 × 10 9   
                 85% 
               
               
                 75 
                 3.5 × 10 8   
                 3 × 10 7   
                 3.3 × 10 9    
                 99% 
               
               
                 80 
                 3.2 × 10 8   
                 0 
                 3.2 × 10 9    
                 100%  
               
               
                 90 
                   3 × 10 8   
                 5 × 10 7   
                 3 × 10 9   
                 99% 
               
               
                 100 
                   3 × 10 8   
                 3 × 10 7   
                 3 × 10 9   
                 99% 
               
               
                 110 
                 3.2 × 10 8   
                 0 
                 3 × 10 9   
                 100%  
               
               
                   
               
             
          
         
       
     
     Goups of ten 8-week old male mice were intraperitoneally administered 1×10 8  cells per animal to study the toxicity of the S and R types of  E. coli  O-157 , Salmonella enteritidis , and  Shigella flexneri . Conserved strains were used for the S types, and strains which had been mutated into 100% R types by  Lactobacillus clearans  were used at that point in time for the R types. The results in Table 16 show that all animals died within 4 days with S type pathogens, whereas none died with the R types except for  Shigella flexneri  which resulted in death by the 7 th  day. 
     
       
         
               
             
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 16 
               
             
             
               
                   
               
               
                 Toxicity of S and R types of  E. coli  O-157, 
               
               
                   Salmonella enteritidis , and  Shigella flexneri  on mice 
               
             
          
           
               
                   
                 S type 
                 R type 
               
               
                   
                   
               
             
          
           
               
                   
                   E. coli  O-157 
                 Death after 3 rd  day 
                 Survived 
               
               
                   
                   
                 following 
                 administration 
               
               
                   
                   
                 administration 
                 (Course) virtually no 
               
               
                   
                   
                 (Course) 
                 movement for 2 days, 
               
               
                   
                   
                 immobilized on 2 nd   
                 matted fur, poor 
               
               
                   
                   
                 day, followed by 
                 complexion 
               
               
                   
                   
                 death 
                 3 rd  to 5 th  day: 
               
               
                   
                   
                   
                 occasionally ate and 
               
               
                   
                   
                   
                 drank 
               
               
                   
                   
                   
                 5 th  to 7 th  day: 
               
               
                   
                   
                   
                 increasingly active 
               
               
                   
                   
                   
                 movement 
               
               
                   
                   
                   
                 After 7 th  day: normal 
               
               
                   
                   
                   
                 activity 
               
               
                   
                 
                   Salmonella 
                 
                 Death after 4 th  day 
                 Survived 
               
               
                   
                 
                   enteritidis 
                 
                 following 
                 administration 
               
               
                   
                   
                 administration 
                 (same course as 
               
               
                   
                   
                 (same course as 
                 O-157) 
               
               
                   
                   
                 above) 
               
               
                   
                 
                   Shigella 
                 
                 Death after 3 rd  day 
                 7 th  day: Death 
               
               
                   
                 
                   flexneri 
                 
                 following 
                 (Course) virtual 
               
               
                   
                   
                 administration 
                 stasis starting on 2 nd   
               
               
                   
                   
                 (same course as 
                 day, but occasional 
               
               
                   
                   
                 above) 
                 drinking 
               
               
                   
                   
                   
                 5 th  day on: 
               
               
                   
                   
                   
                 indisposition and 
               
               
                   
                   
                   
                 subsequent death 
               
               
                   
                   
               
             
          
         
       
     
     The yogurt prepared with  Lactobacillus clearans  described above was orally administered to humans, and blood was analyzed once a month to study the changes in intestinal flora. Cholesterol and triglyceride levels fell about 10% in more than 80% of individuals compared to before administration. 
     Table 17 shows the differences in function between the  Lactobacillus clearans  of the present invention and conventional Lactobacillus strains. 
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 17 
               
             
             
               
                   
               
               
                 Comparison of functions between  Lactobacillus clearans   
               
               
                 and conventional Lactobacillus strains 
               
             
          
           
               
                   
                   
                 Conventional 
               
               
                 Parameter 
                 
                   Lactobacillus clearans 
                 
                 Lactobacillus strains 
               
               
                   
               
               
                 Action against toxic, 
                 degrades, breaks 
                 degrades and reduces 
               
               
                 maladorous intestinal 
                 down, and denatures 
                 malodorous carbon 
               
               
                 putrefying substances 
                 most toxic malodorous 
                 compounds, but not toxic 
               
               
                 such as sulfur, 
                 compounds such as 
                 malodorous compounds 
               
               
                 nitrogen, and 
                 sulfur, nitrogen, and 
                 such as sulfur and 
               
               
                 carbon compounds 
                 carbon compounds to 
                 nitrogen compounds 
               
               
                   
                 reduce them 
               
               
                 Feces deodorization 
                 ++ 
                 − to ± 
               
               
                 Action on beneficial 
                 considerable increases 
                 increases with 
               
               
                 intestinal bacteria 
                   
                 continued ingestion 
               
               
                   
                   
                 in some people 
               
               
                 Bifidobacterium 
                 2 to 10-fold 
                 1 to 3-fold 
               
               
                 Lactobacillus 
                 10 to 100-fold 
                 less than 10-fold 
               
               
                 Action on harmful 
                 harmful bacterial 
                 suppressing action, 
               
               
                 intestinal bacteria 
                 strongly suppressed 
                 but not with most 
               
               
                   
                 with considerable 
               
               
                   
                 increases 
               
               
                 Veillonella 
                 {fraction (1/20)} to {fraction (1/100)} 
                 1 to ⅕ 
               
               
                 Clostridium 
                 {fraction (1/20)} to {fraction (1/100)} 
                 1 to ⅕ 
               
               
                 Antiflatulent action 
                 ++ 
                 − to + 
               
               
                 Nutrient requirements 
                 low to moderate 
                 high 
               
               
                 Intestinal proliferation 
                 + 
                 − to + 
               
               
                 Intestinal stationary 
                 − to + 
                 − 
               
               
                 ability 
               
               
                 Action on coexisting 
                 mutated into non- 
                 no effect 
               
               
                 pathogens 
                 pathogens (S to R 
                 coexistence with 
               
               
                   
                 mutation) 
                 any pathogens 
               
               
                 Salmonella 
                 pathogenicity lost by 
                 succumbs in 
               
               
                   
                 47 th  subculture 
                 competition with 
               
               
                 
                   Shigella flexneri 
                 
                 pathogenicity lost by 
                 pathogens during 
               
               
                   
                 108 th  subculture 
                 subculture 
               
               
                   E. coli  (O-157) 
                 pathogenicity lost by 
               
               
                   
                 18 th  subculture 
               
               
                 Cholesterol and 
                 weak, but able 
                 none 
               
               
                 triglycerides 
                 to reduce 
               
               
                   
               
             
          
         
       
     
       Enterococcus faecalis  is a group of bacteria constituting the intestinal flora and belongs to the group of lactic acid bacteria, normally occurring in an amount of about 1×10 7  cells per gram feces. They appear in the form of two spherical or ovoid shapes, or in the form of short chains. They are gram positive cocci with potent resistance to heat, drying, chlorine, gallic acid, and the like, and grow in a wider temperature range of 10 to 45° C. compared to common streptococci. Although there are some pathogenic strains, they are nontoxic. The viable cells act as antiflatulents, and have been commercially available for more than 10 years in Japan. They have been proven to be nontoxic when taken orally. Apart from such commercial products, viable and killed cells of some strains have recently been shown to effectively lower blood cholesterol and triglycerides. They thus hold promise in the prevention or treatment of typically related adult diseases (diseases stemming from life style habits), such as hyperlipidemia, hypertension, and arteriosclerosis. The  Enterococcus faecalis  referred to in the present invention indicates strains having such functions. 
     The following is an example of a method for preparing viable cells of  Enterococcus faecalis  for use in the present invention. Specifically, cells which have been cultured and centrifuged by a common method are suspended in physiological saline, washed, centrifuged again, and collected. They may be lyophilized using soluble starch as a preservative. As an example of a method for preparing killed cells of  Enterococcus faecalis , soluble starch and the washed and centrifuged cells obtained when the aforementioned viable cells have been collected can be treated for 30 minutes in 100° C. hot water and then lyophilized. 
     Although  Enterococcus faecalis  has no in vitro capacity for reducing sodium sulfide (Na 2 S.9H 2 O) or ammonia, it does have a weak deodorizing capacity against 10-fold dilution of feces. Table 18 gives the deodorizing capacity of two typical strains of  Enterococcus faecalis , namely, FERM BP-7230 and FERM BP-7231. The test was the same as that for  Lactobacillus clearans , and the criteria were the same as those given in Table 1. 
     
       
         
               
             
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 18 
               
             
             
               
                   
               
               
                 Deodorizing capacity of  Enterococcus faecalis  (in vitro) 
               
             
          
           
               
                   
                 Odor (average value by panel) 
                   
               
             
          
           
               
                 Inoculum 
                   
                 High pressure 
                   
               
               
                 FERM BP-No. 
                   
                 sterilization 
                 Unsterilized 
               
               
                   
               
               
                 7230 
                 Viable cells 
                 3.0 
                 3.5 
               
               
                   
                 Killed cells 
                 4.0 
                 4.0 
               
               
                 7231 
                 Viable cells 
                 3.2 
                 3.5 
               
               
                   
                 Killed cells 
                 4.0 
                 4.5 
               
               
                   
               
             
          
         
       
     
     The two typical strains of Enterococcus faecalis were orally administered in amounts of 1×10 9  cells/person per day. Feces odor prior to administration was rated as 100. Feces odor was determined 20 to 30 days, 50 to 60 days, and 80 to 90 days following ingestion, with the results given in Table 19. Table 19 shows that odor was reduced by  Enterococcus faecalis , albeit weakly. 
     
       
         
               
             
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 19 
               
             
             
               
                   
               
               
                 feces odor during ingestion of  Enterococcus faecalis   
               
             
          
           
               
                   
                 Odor 
                   
               
               
                   
                 prior 
                 Odor after ingestion 
               
             
          
           
               
                 Inoculum 
                   
                 to 
                 20 to 30 
                 50 to 60 
                 80 to 90 
               
               
                 FERM BP-No. 
                   
                 ingestion 
                 days 
                 days 
                 days 
               
               
                   
               
               
                 7230 
                 Viable cells 
                 100 
                 90 
                 80 
                 65 
               
               
                   
                 Killed cells 
                 100 
                 90 
                 83 
                 77 
               
               
                 7231 
                 Viable cells 
                 100 
                 85 
                 75 
                 70 
               
               
                   
                 Killed cells 
                 100 
                 90 
                 80 
                 75 
               
               
                   
               
             
          
         
       
     
     Viable and killed cells of two typical strains of  Enterococcus faecalis , namely, FERM BP-7230 and BP-7231, were prepared and mixed in equal parts viable and killed cells to produce a preparation. The preparation was taken in an amount of 1×10 9  cells/day/person for 6 months, and the changes in the cell count of the intestinal flora were determined over time by measuring the changes in the cell count (cells/1 g feces)of Bifidobacterium and Lactobacillus (except for  Lactobacillus clearans ) which are known as typical beneficial bacteria, as well as of typical harmful bacteria such as Veillonella and  Clostridium perfringens . Table 20 shows the results of oral administration to 10 healthy individuals. Table 20 shows that the viable cells had a slightly higher rate of improvement than the killed cells. The results were far lower than those of  Lactobacillus clearans , however. 
     
       
         
               
             
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 20 
               
             
             
               
                   
               
               
                 Effect of oral administration of  Enterococcus   
               
               
                   faecalis  preparation on intestinal flora in healthy 
               
               
                 individuals 
               
             
          
           
               
                   
                 Administration of 
                 Administration of 
               
               
                   
                 viable cells 
                 Killed cells 
               
             
          
           
               
                   
                 Cell 
                   
                 Cell 
                   
               
               
                   
                 count 
                   
                 count 
               
               
                   
                 before 
                 Cell count 
                 before 
                 Cell count 
               
               
                   
                 admin. 
                 after 6 months 
                 admin. 
                 after 6 months 
               
               
                   
                   
               
             
          
           
               
                 Beneficial 
                 Bifidoba 
                 1.2 × 10 10   
                 1.5 × 10 10   
                 25% 
                 1.5 × 10 10   
                 1.8 × 10 10   
                 20% 
               
               
                 bacteria 
                 cterium 
                   
                   
                 increase 
                   
                   
                 increase 
               
               
                   
                 Lactobac 
                 1.8 × 10 7   
                   3 × 10 7   
                 67% 
                   2 × 10 7   
                   3 × 10 7   
                 50% 
               
               
                   
                 illus 
                   
                   
                 increase 
                   
                   
                 increase 
               
               
                 Harmful 
                 Clostrid 
                   1 × 10 5   
                 0.7 × 10 5   
                 30% 
                 1.2 × 10 5   
                 0.9 × 10 5   
                 25% 
               
               
                 bacteria 
                 ium 
                   
                   
                 decrease 
                   
                   
                 decrease 
               
               
                   
                 Veillone 
                 5.5 × 10 5   
                 4.4 × 10 5   
                 20% 
                   6 × 10 5   
                   5 × 10 5   
                 17% 
               
               
                   
                 lla 
                   
                   
                 decrease 
                   
                   
                 decrease 
               
               
                   
               
             
          
         
       
     
     Mixed cultures of  Escherichia coli  O-157 , Salmonella enteritidis , and  Shigella flexneri  were performed to study the action of  Enterococcus faecalis  against pathogens. The ratios in which the pathogens mutated from S to R types were virtually the same as spontaneously occurring ratios. Pathogen cell counts gradually decreased with each subculture, until  Salmonella enteritidis  had disappeared by the 25 th  subculture,  Shigella flexneri  had disappeared by the 33 rd  subculture, and  E. coli  O-157 had disappeared by the 35 th  culture. Although this suggested that the growth of  Enterococcus faecalis  overcame pathogens, the potential production of some physiologically active substances suppressing pathogen growth could not be ruled out. 
     The aforementioned viable and killed cell preparations of  Enterococcus faecalis  were added, in amounts of 1×10 9  cells/1 gram feed each, to feed. Mice raised for 3 months on this and mice raised on conventional feed alone were compared for serum triglycerides and cholesterol. The mice were divided into groups of 10, and were allowed to feed freely. The triglycerides and cholesterol were measured by means of an E-Test Wako and C-Test Wako, respectively. The mean level for the control mice was 100%. The results are given in Table 21. Table 21 shows that mice continually ingesting either viable or killed cells had a 20 to 40% reduction in levels for both parameters, leaving no room for doubt as to their effectiveness. 
     
       
         
               
             
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 21 
               
             
             
               
                   
               
               
                 Effect of oral administration of  Enterococcus faecalis  preparation 
               
               
                 on serum triglyceride and cholesterol levels in mice 
               
             
          
           
               
                 Inoculum 
                 Triglycerides 
                   
                 Cholesterol 
               
             
          
           
               
                 FERM BP-No. 
                 Viable cells 
                 Killed cells 
                 Viable cells 
                 Killed cells 
               
               
                   
               
               
                 7230 
                 72% 
                 80% 
                 65% 
                 70% 
               
               
                 7231 
                 68% 
                 65% 
                 60% 
                 57% 
               
               
                   
               
             
          
         
       
     
     Fifteen 8-week old spontaneous hypertensive rats (SHR) were divided into 3 groups. The groups were fed feed containing viable and killed cell preparations of  Enterococcus faecalis  in amounts of 1×10 9  cells/gram feed. The control group was not administered any cell preparation. The animals were raised for 3 months. Table 22 gives the blood pressure after 3 months. Table 22 shows an approximately 10% decrease in blood pressure. 
     
       
         
               
             
               
               
               
             
               
               
               
               
               
             
               
               
               
             
           
               
                 TABLE 22: 
               
             
             
               
                   
               
               
                 effects of oral administration of  Enterococcus   
               
               
                   faecalis  preparation on rat blood pressure 
               
             
          
           
               
                   
                 Blood 
                 Blood pressure 
               
               
                 Inoculum 
                 pressure 
                 after administration 
               
             
          
           
               
                 FERM BP- 
                 before 
                   
                   
                 Percent 
               
               
                 No. 
                 admin. 
                 Viable cells 
                 Killed cells 
                 decrease 
               
               
                   
               
               
                 7230 
                 205 mmHg 
                 185 mmHg 
                 178 mmHg 
                 9.7 to 13%  
               
               
                 7231 
                 208 mmHg 
                 190 mmHg 
                 185 mmHg 
                 7.3 to 9.7% 
               
             
          
           
               
                 Control 
                 202 mmHg 
                 210 mmHg 
               
               
                   
               
             
          
         
       
     
    
    
     EXAMPLES 
     The combined action of  Lactobacillus clearans  and  Enterococcus faecalis  allowed surprisingly effective results to be rapidly obtained. Manufacturing methods and examples of the preparations in the present invention are described below, but the scope of the present invention is not limited by these manufacturing examples and working examples. 
     Manufacturing Example 1 
     Production of  Lactobacillus clearans  preparation: 10 L medium comprising 5 g meat extract, 5 g peptone, 3 g sodium acetate, 1 mL ammonia water, 10 g glucose, 0.5 g cystine, and 2 g yeast extract per liter medium was inoculated with  Lactobacillus clearans  for 72 hours of anaerobic culture at 37° C. The resulting culture was centrifuged, giving 10 g biomass. This was washed with 500 mL physiological saline and centrifuged, twice. The washed biomass was introduced into 500 mL solution comprising 50 g skim milk, 30 g trehalose, and 0.5 g taurine, and was thoroughly stirred. The mixture was lyophilized by a common method, giving 82.5 g cell preparation (3×10 9  cell/g). This was mixed with 330 g thoroughly dried skim milk, giving a  Lactobacillus clearans  preparation containing 5×10 8  viable cells per gram. 
     Manufacturing Example 2 
     Production of  Enterococcus faecalis  viable cell preparation: 10 L medium comprising 5 g meat extract, 5 g peptone, 2 g sodium chloride, 2 g yeast extract, and 10 g glucose per liter medium was inoculated with  Enterococcus faecalis  for 72 hours of aerobic culture at 37° C. The resulting culture was centrifuged, giving 16 g biomass. This was washed with 800 mL physiological saline and centrifuged, twice. The washed biomass was introduced into 500 mL solution comprising 20 g skim milk, 30 g soluble starch, and 0.5 g sodium glutamate, and was thoroughly stirred. The mixture was lyophilized by a common method, giving 54 g cell preparation (5×10 9  cell/g). This was mixed with 486 g thoroughly dried soluble starch, giving a  Enterococcus faecalis  viable cell preparation containing 5×10 8  viable cells per gram. 
     Manufacturing Example 3 
     Production of  Enterococcus faecalis  killed cell preparation: The washed biomass obtained in Manufacturing Example 2 was suspended in 500 mL physiological saline, 50 g soluble starch was then introduced, the solution was thermally sterilized for 15 minutes at 110° C., and the suspension was lyophilized by a common method, giving 53 g cell preparation (approximately 5×10 9  cells/g). Killed cell preparations can also be produced by rupturing the cells ultrasonically or the like. 
     Manufacturing Example 4 
     Production of  Lactobacillus clearans  and  Enterococcus faecalls  mixture preparation: The  Lactobacillus clearans  preparation produced in Manufacturing Example 1 and the  Enterococcus faecalis  viable cell preparation produced in Manufacturing Example 2 or the  Enterococcus faecalis  killed cell preparation produced in Manufacturing Example 3 were mixed in equal parts to produce a mixture preparation. The preparation in this case contained  Lactobacillus clearans  in an amount of 2.5×10 8  cells/g and  Enterococcus faecalis  in an amount of 2.5×10 8  cells/g, but the ratio between the  Lactobacillus clearans  and  Enterococcus faecalis  preparations can be varied during the manufacturing process to produce mixtures containing any desired cell count. The preparation may be in the form of powders, granules, capsules, or other common formulations with a suitable excipient. 
     Example 1 
     15 mL of a 10-fold dilution of feces was introduced into test tubes (18×180 mm). The test tubes were then inoculated with sample bacteria 1) that had been sterilized at high pressure for 15 minutes at 120° C. and 2) that had not been sterilized. The test tubes were stoppered with rubber plugs for 72 hours of anaerobic culture at 37° C. 1 mL air was then drawn by syringe from inside the test tubes and injected into 5 L odor bags for olfactory tests by a panel of 6 individuals. The odor was assessed based on the feces odor criteria given in Table 1. For  Enterococcus faecalis  killed cell preparations, 0.1 g of the preparation produced in Manufacturing Example 3 was added to the test tubes. The test results in Table 23 show that the feces odor was considerably weakened, particularly in the case of feces diluent treated with samples sterilized at high pressure. This will be evident in a comparison with the results for test batches inoculated with  Lactobacillus clearans  alone in Table 2. 
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 23 
               
             
             
               
                   
               
               
                 test of deodorization with mixture of 
               
               
                   Lactobacillus clearans  and  Enterococcus faecalis  (1) 
               
             
          
           
               
                   
                 Cells used 
                   
                 In vitro test 
                   
               
             
          
           
               
                   
                 
                   Lactobacillus 
                 
                 
                   Enterococcus 
                 
                 Sterilized 
                   
               
               
                   
                 
                   clearans 
                 
                 
                   faecalis 
                 
                 at high 
               
               
                   
                 FERM BP-No. 
                 FERM BP-No. 
                 pressure 
                 Unsterilized 
               
               
                   
                   
               
               
                   
                 6973 
                 7230 
                 1.2 
                 1.5 
               
               
                   
                   
                 (viable) 
               
               
                   
                   
                 7230 
                 1.3 
                 1.7 
               
               
                   
                   
                 (killed) 
               
               
                   
                   
               
             
          
         
       
     
     Example 2 
     A combination of 1 g  Enterococcus faecalis  (FERM BP-7230) viable cell preparation or 1 g  Enterococcus faecalis  (FERM BP-7230) killed cell preparation and 100 mL yogurt prepared with  Lactobacillus clearans  FERM BP-6973 in medium comprising 100 g skim milk, 50 g sucrose, and 2 g agar per liter was continuously administered to 10 individuals, and the feces odor was continuously determined 20 to 30 days, 50 to 60 days, and 80 to 90 days after administration. The results were determined by introducing 0.5 g feces samples from the 10 individuals into 20 L odor bags, which were stored at room temperature for evaluation of the odor by a panel of 6 individuals after 30 minutes. The average feces odor for 5 individuals ingesting no yogurt was rated as 100. The mean values in Table 24 show that oral administration increasingly weakened feces odor over time. After 90 days, the odor was considerably diminished and was no longer disagreeable. The appreciable effects will be evident when compared with the results obtained with the administration of  Lactobacillus clearans  alone in Table 3. 
     
       
         
               
             
               
               
             
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 24 
               
             
             
               
                   
               
               
                 test of deodorization with mixture of 
               
               
                   Lactobacillus clearans  and  Enterococcus faecalis  (2) 
               
             
          
           
               
                 Cells used 
                   
               
             
          
           
               
                 
                   Lactobacillus 
                 
                 
                   Enterococcus 
                 
                 In vitro test 
               
             
          
           
               
                 
                   clearans 
                 
                 
                   faecalis 
                 
                 20 to 30 
                 50 to 60 
                 80 to 90 
               
               
                 FERM BP-No. 
                 FERM BP-No. 
                 days 
                 days 
                 days 
               
               
                   
               
               
                 6973 
                 7230 
                 20 
                 10 
                 5 
               
               
                   
                 (viable) 
               
               
                   
                 7230 
                 33 
                 16 
                 8 
               
               
                   
                 (killed) 
               
               
                   
               
             
          
         
       
     
     Example 3 
     In the same manner as in Example 1, test tubes of 10-fold feces diluent were inoculated with sample bacteria 1) that had been sterilized at high pressure for 15 minutes at 120° C. and 2) that had not been sterilized. The test tubes were stoppered with rubber plugs for 72 hours of anaerobic culture at 37° C. The cultures were then centrifuged, 5 mL supernatant was collected from each and diluted 10-fold, and the concentrations of free sulfur ions (S 2+ ) and ammonium ions (NH 4   + ) were determined in the resulting 50 mL solution. The concentration of lower fatty acids was also determined by gas chromatography. Tables 25 and 26 give the measured results. The tables show that typical substances in feces were reduced at a high rate of 70 to 90% by both sterilized and unsterilized samples. The rate of reduction will be evident in a comparison with the results obtained upon inoculation with  Lactobacillus clearans  alone in Tables 6 and 7. 
     
       
         
               
             
               
               
             
               
               
               
             
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 25 
               
             
             
               
                   
               
               
                 Test on decrease in toxic malodorous substances by mixture of  Lactobacillus clearans   
               
               
                 and  Enterococcus faecalis  (1) 
               
               
                 (sterilized feces dilution) 
               
             
          
           
               
                 Bacteria 
                   
               
             
          
           
               
                 
                   Lactobacillus 
                 
                 
                   Enterococcus 
                 
                 Results of study of chemical analysis 
               
             
          
           
               
                 
                   clearans 
                 
                 
                   faecalis 
                 
                 Before culture (ppm) 
                 After culture (ppm) 
               
             
          
           
               
                 FERM BP-No. 
                 FERM BP-No. 
                 S 2+   
                 NH 4   +   
                 Fatty acids 
                 S 2+   
                 NH 4   +   
                 Fatty acids 
               
               
                   
               
               
                 6973 
                 7230 
                 30 
                 350 
                 2800 
                 3 
                 70 
                 500 
               
               
                   
                 (viable) 
                   
                   
                   
                 90% decrease 
                 80% decrease 
                 82% decrease 
               
               
                   
                 7230 
                 30 
                 350 
                 2800 
                 4 
                 80 
                 620 
               
               
                   
                 (killed) 
                   
                   
                   
                 87% decrease 
                 77% decrease 
                 78% decrease 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
             
               
               
               
             
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 26 
               
             
             
               
                   
               
               
                 Test on decrease in toxic malodorous substances by mixture of  Lactobacillus clearans   
               
               
                 and  Enterococcus faecalis  (2) 
               
               
                 (unsterilized feces dilution) 
               
             
          
           
               
                 Bacteria 
                   
               
             
          
           
               
                 
                   Lactobacillus 
                 
                 
                   Enterococcus 
                 
                 Results of study of chemical analysis 
               
             
          
           
               
                 
                   clearans 
                 
                 
                   faecalis 
                 
                 Before culture (ppm) 
                 After culture (ppm) 
               
             
          
           
               
                 FERM BP-No. 
                 FERM BP-No. 
                 S 2+   
                 NH 4   +   
                 Fatty acids 
                 S 2+   
                 NH 4   +   
                 Fatty acids 
               
               
                   
               
               
                 6973 
                 7230 
                 50 
                 370 
                 3500 
                  9 
                 65 
                  870 
               
               
                   
                 (viable) 
                   
                   
                   
                 82% decrease 
                 82% decrease 
                 75% decrease 
               
               
                   
                 7230 
                 50 
                 370 
                 3500 
                 12 
                 80 
                 1020 
               
               
                   
                 (killed) 
                   
                   
                   
                 76% decrease 
                 78% decrease 
                 76% decrease 
               
               
                   
               
             
          
         
       
     
     Example 4 
     1 g  Lactobacillus clearans  preparation (5×10 8  cells/g) comprising a mixture of equal parts of three typical strains of  Lactobacillus clearans , specifically, FERM BP-6972, FERM BP-6971, FERM BP-6973, and 1 g  Enterococcus faecalis  viable cell preparation (5×10 8  cells/g) of  Enterococcus faecalis  FERM BP-7230 or 1 g  Enterococcus faecalis  killed cell preparation (5×10 8  cells/g) of  Enterococcus faecalis  FERM BP-7231 were continuously administered for 6 months to 20 healthy individuals and 20 constitutionally weak individuals. The intestinal flora were monitored over time, with the results given in Tables 27 and 28. The tables show that the oral administration of  Enterococcus faecalis  viable or killed cells mixed with  Lactobacillus clearans  more rapidly increased beneficial bacterial and decreased harmful bacteria than when  Lactobacillus clearans  was administered alone, as shown in Tables 8 and 9. That is, improvements were about 25% faster in healthy individuals, with a 10 to 50% higher beneficial bacterial cell count and a 20 to 40% lower harmful bacterial cell count. In constitutionally weak individuals, improvements were about 30% faster, with a 10 to 30% higher beneficial bacterial cell count and a 20 to 50% lower harmful bacterial cell count. It may thus be concluded that improvement in the intestinal flora was more effective in constitutionally weak individuals. 
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 27 
               
             
             
               
                   
               
               
                 effects of mixture of  Lactobacillus clearans  and  Enterococcus faecalis  on intestinal 
               
               
                 flora (healthy individuals) 
               
             
          
           
               
                   
                   
                   
                 Cell count 
                   
               
               
                 
                   Lactobacillus 
                 
                 
                   Enterococcus 
                 
                   
                 before 
                 Change in cell count after administration 
               
             
          
           
               
                 
                   clearans 
                 
                 
                   faecalis 
                 
                   
                 admin. 
                 1 month 
                 2 months 
                 3 months 
                 6 months 
               
               
                   
               
               
                 FERM 
                 FERM 
                 Bifidobacterium 
                 1.25 × 10 10   
                 1.65 × 10 10   
                 2.0 × 10 10   
                  2.3 × 10 10   
                  2.8 × 10 10   
               
               
                 BP-6972 
                 BP-7230 
                 Lactobacillus 
                  2.4 × 10 7   
                  4.0 × 10 7   
                 6.5 × 10 7   
                   10 × 10 7   
                   15 × 10 7   
               
               
                 6971 
                 (viable 
                 Clostridium 
                  1.2 × 10 5   
                  0.8 × 10 5   
                 0.5 × 10 5   
                  0.3 × 10 5   
                  0.1 × 10 5   
               
               
                 6973 
                 cells) 
                 Veillonella 
                  5.0 × 10 5   
                  4.0 × 10 5   
                 2.5 × 10 5   
                  2.0 × 10 5   
                  1.0 × 10 5   
               
               
                   
                 FERM 
                 Bifidobacterium 
                 1.25 × 10 10   
                 1.60 × 10 10   
                 1.0 × 10 10   
                 2.20 × 10 10   
                 2.65 × 10 10   
               
               
                   
                 BP-7231 
                 Lactobacillus 
                  2.4 × 10 7   
                  3.5 × 10 7   
                 5.8 × 10 7   
                  8.5 × 10 7   
                   14 × 10 7   
               
               
                   
                 (killed 
                 Clostridium 
                  1.2 × 10 5   
                  1.0 × 10 5   
                 0.7 × 10 5   
                  0.4 × 10 5   
                 0.13 × 10 5   
               
               
                   
                 cells) 
                 Veillonella 
                  5.0 × 10 5   
                  4.0 × 10 5   
                 3.0 × 10 5   
                  2.3 × 10 5   
                  1.2 × 10 5   
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
               
             
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 28 
               
             
             
               
                   
               
               
                 effects of mixture of  Lactobacillus clearans  and  Enterococcus faecalis  on intestinal 
               
               
                 flora (constitutionally weak individuals) 
               
             
          
           
               
                   
                   
                   
                 Cell count 
                   
               
               
                 
                   Lactobacillus 
                 
                 
                   Enterococcus 
                 
                   
                 before 
                 Change inn cell count after administration 
               
             
          
           
               
                 
                   clearans 
                 
                 
                   faecalis 
                 
                   
                 admin. 
                 1 month 
                 2 months 
                 3 months 
                 6 months 
               
               
                   
               
               
                 FERM 
                 FERM 
                 Bifidobacterium 
                  4.5 × 10 9   
                  5.5 × 10 9   
                  7.0 × 10 9   
                  8.0 × 10 9   
                 10.0 × 10 9   
               
               
                 BP-6972 
                 BP-7230 
                 Lactobacillus 
                  1.0 × 10 7   
                  1.2 × 10 7   
                  1.8 × 10 7   
                  3.0 × 10 7   
                  5.0 × 10 7   
               
               
                 6971 
                 (viable 
                 Clostridium 
                  4.5 × 10 5   
                  2.5 × 10 5   
                  1.8 × 10 5   
                  1.4 × 10 5   
                  1.0 × 10 5   
               
               
                 6973 
                 cells) 
                 Veillonella 
                  2.2 × 10 6   
                  1.6 × 10 6   
                  1.0 × 10 6   
                 0.65 × 10 6   
                 0.38 × 10 6   
               
               
                   
                 FERM 
                 Bifidobacterium 
                  4.5 × 10 9   
                  5.0 × 10 9   
                  6.0 × 10 9   
                  7.0 × 10 9   
                  9.0 × 10 9   
               
               
                   
                 BP-7231 
                 Lactobacillus 
                  1.0 × 10 7   
                  1.1 × 10 7   
                  1.5 × 10 7   
                  2.3 × 10 7   
                  4.5 × 10 7   
               
               
                   
                 (killed 
                 Clostridium 
                  4.5 × 10 5   
                  2.8 × 10 5   
                  2.0 × 10 5   
                  1.5 × 10 5   
                  1.0 × 10 5   
               
               
                   
                 cells) 
                 Veillonella 
                 2.20 × 10 6   
                 1.70 × 10 6   
                 1.20 × 10 6   
                 0.75 × 10 6   
                  0.4 × 10 6   
               
               
                   
               
             
          
         
       
     
     Example 5 
     Viable or killed cells of  Enterococcus faecalis  were added to mixed cultures of  Lactobacillus clearans, E.coli  O-157 , Salmonella enteritidis , or  Shigella flexneri  for anaerobic culture at 37° C., and subcultures were repeated every 72 hours, at which point the plates were diluted and smeared to monitor the changes in cell count and the ratio of S-R mutation of  E. coli  O-157 , Salmonella enteritidis , and  Shigella flexneri . The results are given in Tables 29, 30, 31, 32, 33, and 34. The FERM BP-6973 strain of  Lactobacillus clearans  was used, as was the FERM BP-7231 strain of  Enterococcus faecalis . The aforementioned  Enterococcus faecalis  killed cell preparation was added in an amount of 1 g per L medium. The medium composition used for cultures comprised 10 g meat extract, 10 g peptone, 2 g glucose, 2 g NaCl, and 1 g CaCO 3  per liter, with the pH adjusted to 7.2, and was sterilized at high pressure for 15 minutes at 120° C. The tables below reveal that the inoculation of pathogens such as  E. coli  O-157 , Salmonella enteritidis , and  Shigella flexneri  with  Lactobacillus clearans  and  Enterococcus faecalis  viable cells rapidly diminished the cell counts of such pathogens and resulted in their mutation to R types over the course of subculturing.  E. coli  O-157 mutated 100% to R type by the 13th subculture, and disappeared by the 15th subculture.  Salmonella enteritidis  and  Shigella flexneri  disappeared before mutating 100% to R type by the 13th and 15th subcultures, respectively. When  Lactobacillus clearans  was used by itself, as shown in Tables 11, 13, and 15, these pathogens mutated into R types over the course of numerous subcultures, but the bacteria maintained a constant cell count of 1 to 3×10 9  cells/mL, without disappearing. Although the addition of  Lactobacillus clearans  and  Enterococcus faecalis  killed cells to the aforementioned pathogens showed the same tendencies as that prevailing with the inoculation of viable cells, the mutation to R types was slower than that prevailing with the inoculation of viable cells, and the cell counts diminished but did not disappear. Nevertheless, the number of subcultures resulting in 100% mutation to R types was 15 with  E. coli  O-157, 30 with  Salmonella enteritidis , and 15 with  Shigella flexneri , which were far more rapid then when  Lactobacillus clearans  was used as inoculum on its own. 
     
       
         
               
             
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 29 
               
             
             
               
                   
               
               
                 effects of  Lactobacillus clearans  (FERM BP-6973) 
               
               
                 and  Enterococcus faecalis  (FERM BP-7231) viable cells on 
               
               
                   E. coli  O-157 
               
             
          
           
               
                 Number 
                 
                   L. clearans 
                 
                 
                   E. faecalis 
                 
                   E. coli  O-157 
               
             
          
           
               
                 Subcul- 
                 FERM BP-6973 
                 FERM BP-7231 
                 S type 
                 R type 
                 Ratio 
               
               
                 ture 
                 cells/mL 
                 cells/mL 
                 cells/mL 
                 cells/mL 
                 of R type 
               
               
                   
               
               
                 1 
                   1 × 10 9   
                   2 × 10 9   
                   2 × 10 9   
                 0 
                 0 
               
               
                 3 
                   1 × 10 9   
                 2.2 × 10 9   
                 1.5 × 10 9   
                 0 
                 0 
               
               
                 5 
                 1.2 × 10 9   
                 2.4 × 10 9   
                   1 × 10 9   
                 2 × 10 8   
                 20% 
               
               
                 7 
                   1 × 10 9   
                   2 × 10 9   
                   2 × 10 8   
                 3 × 10 8   
                 60% 
               
               
                 9 
                   8 × 10 8   
                 2.4 × 10 9   
                   1 × 10 8   
                 2 × 10 8   
                 67% 
               
               
                 11  
                   1 × 10 9   
                 2.5 × 10 9   
                   0 
                 1 × 10 8   
                 100%  
               
               
                 13  
                 1.2 × 10 9   
                 2.3 × 10 9   
                   0 
                 4 × 10 7   
                 100%  
               
               
                 15  
                 1.2 × 10 9   
                 2.5 × 10 9   
                   0 
                 0 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 30 
               
             
             
               
                   
               
               
                 effects of  Lactobacillus clearans  (FERM BP-6973) 
               
               
                 and  Enterococcus faecalis  (FERM BP-7231) viable cells on 
               
               
                 
                   Salmonella enteritidis 
                 
               
             
          
           
               
                 Number 
                 
                   L. clearans 
                 
                 
                   E. faecalis 
                 
                   E. coli  O-157 
               
             
          
           
               
                 Subcul- 
                 FERM BP-6973 
                 FERM BP-7231 
                 S type 
                 R type 
                 Ratio 
               
               
                 ture 
                 cells/mL 
                 cells/mL 
                 cells/mL 
                 cells/mL 
                 of R type 
               
               
                   
               
               
                 1 
                   1 × 10 9   
                 2.5 × 10 9   
                 1.2 × 10 9   
                 0 
                   
               
               
                 3 
                 1.2 × 10 9   
                 2.5 × 10 9   
                   1 × 10 9   
                 2 × 10 7   
                  2% 
               
               
                 5 
                 1.5 × 10 9   
                 2.2 × 10 9   
                   8 × 10 8   
                 5 × 10 8   
                 38% 
               
               
                 7 
                 1.3 × 10 9   
                   3 × 10 9   
                   1 × 10 8   
                 1 × 10 8   
                 50% 
               
               
                 9 
                   1 × 10 9   
                   3 × 10 9   
                   7 × 10 7   
                 8 × 10 7   
                 53% 
               
               
                 11  
                   1 × 10 9   
                 3.2 × 10 9   
                   3 × 10 7   
                 5 × 10 7   
                 63% 
               
               
                 13  
                   8 × 10 8   
                   3 × 10 9   
                   0 
                 0 
               
               
                 15  
                   1 × 10 9   
                 2.8 × 10 9   
                   0 
                 0 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 31 
               
             
             
               
                   
               
               
                 effects of  Lactobacillus clearans  (FERM BP-6973) 
               
               
                 and  Enterococcus faecalis  (FERM BP-7231) viable cells on 
               
               
                 
                   Shigella flexneri 
                 
               
             
          
           
               
                 Number 
                 
                   L. clearans 
                 
                 
                   E. faecalis 
                 
                 
                   Shigella flexneri 
                 
               
             
          
           
               
                 Subcul- 
                 FERM BP-6973 
                 FERM BP-7231 
                 S type 
                 R type 
                 Ratio 
               
               
                 ture 
                 cells/mL 
                 cells/mL 
                 cells/mL 
                 cells/mL 
                 of R type 
               
               
                   
               
               
                 1 
                 1.2 × 10 9   
                   2 × 10 9   
                 1.5 × 10 9   
                 0 
                 0 
               
               
                 3 
                   1 × 10 9   
                 2.2 × 10 9   
                   1 × 10 9   
                 0 
                 0 
               
               
                 5 
                   7 × 10 8   
                 2.5 × 10 9   
                   4 × 10 8   
                 1 × 10 8   
                 20% 
               
               
                 7 
                   5 × 10 8   
                 2.8 × 10 9   
                   2 × 10 8   
                 2 × 10 8   
                 50% 
               
               
                 9 
                   7 × 10 8   
                 2.5 × 10 9   
                   1 × 10 8   
                 2 × 10 8   
                 67% 
               
               
                 11  
                   5 × 10 8   
                 2.5 × 10 9   
                   3 × 10 7   
                 6 × 10 7   
                 67% 
               
               
                 13  
                   5 × 10 8   
                 2.7 × 10 9   
                   2 × 10 6   
                 6 × 10 6   
                 70% 
               
               
                 15  
                   6 × 10 8   
                 2.5 × 10 9   
                   0 
                 0 
               
               
                 17  
                   8 × 10 8   
                   3 × 10 9   
                   0 
                 0 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 32 
               
             
             
               
                   
               
               
                 effects of  Lactobacillus clearans  (FERM BP-6973) 
               
               
                 and  Enterococcus faecalis  (FERM BP-7231) killed cells on 
               
               
                 
                   E. coli O-157 
                 
               
             
          
           
               
                   
                 
                   L. clearans 
                 
                   E. coli  O-157 
               
             
          
           
               
                 Number 
                 FERM BP-6973 
                 S type 
                 R type 
                 Ratio 
               
               
                 subculture 
                 cells/mL 
                 cells/mL 
                 cells/mL 
                 of R type 
               
               
                   
               
             
          
           
               
                 1 
                 1.8 × 10 9   
                 2.5 × 10 9   
                 0 
                 0 
               
               
                 3 
                   2 × 10 9   
                   2 × 10 9   
                 0 
                 0 
               
               
                 5 
                   2 × 10 9   
                 1.5 × 10 9   
                   5 × 10 8   
                 25% 
               
               
                 7 
                 1.8 × 10 9   
                   8 × 10 9   
                   1 × 10 9   
                 56% 
               
               
                 8 
                 1.5 × 10 9   
                   5 × 10 8   
                   8 × 10 8   
                 62% 
               
               
                 11 
                 1.5 × 10 9   
                   1 × 10 8   
                   5 × 10 8   
                 82% 
               
               
                 13 
                 1.8 × 10 9   
                   5 × 10 7   
                 4.5 × 10 8   
                 90% 
               
               
                 15 
                   2 × 10 9   
                 0 
                   2 × 10 8   
                 100%  
               
               
                 18 
                   2 × 10 9   
                 0 
                   2 × 10 8   
                 100%  
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 33 
               
             
             
               
                   
               
               
                 effects of  Lactobacillus clearans  (FERM BP-6973) 
               
               
                 and  Enterococcus faecalis  (FERM BP-7231) killed cells on 
               
               
                 
                   Salmonella enteritidis 
                 
               
             
          
           
               
                   
                 
                   L. clearans 
                 
                 
                   Salmonella enteritidis 
                 
               
             
          
           
               
                 Number 
                 FERM BP-6973 
                 S type 
                 R type 
                 Ratio 
               
               
                 subculture 
                 cells/mL 
                 cells/mL 
                 cells/mL 
                 of R type 
               
               
                   
               
             
          
           
               
                 1 
                 1.5 × 10 9   
                   3 × 10 9   
                 0 
                 0 
               
               
                 3 
                 1.2 × 10 9   
                 2.5 × 10 9   
                 0 
                 0 
               
               
                 5 
                 1.2 × 10 9   
                 1.5 × 10 9   
                   7 × 10 8   
                 32% 
               
               
                 7 
                   1 × 10 9   
                 1.2 × 10 9   
                   1 × 10 8   
                 45% 
               
               
                 9 
                   1 × 10 9   
                   8 × 10 8   
                 1.2 × 10 9   
                 60% 
               
               
                 11 
                   8 × 10 8   
                   5 × 10 8   
                   8 × 10 8   
                 61% 
               
               
                 13 
                   8 × 10 8   
                   5 × 10 8   
                   5 × 10 8   
                 50% 
               
               
                 15 
                   5 × 10 8   
                   2 × 10 8   
                   8 × 10 8   
                 80% 
               
               
                 20 
                   7 × 10 8   
                   1 × 10 8   
                   9 × 10 8   
                 90% 
               
               
                 25 
                   5 × 10 8   
                   2 × 10 8   
                   1 × 10 9   
                 83% 
               
               
                 30 
                   4 × 10 8   
                 0 
                 1.8 × 10 9   
                 100%  
               
               
                 35 
                   5 × 10 8   
                 0 
                 1.5 × 10 9   
                 100%  
               
               
                 40 
                   6 × 10 8   
                 0 
                   1 × 10 9   
                 100%  
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 34 
               
             
             
               
                   
               
               
                 effects of  Lactobacillus clearans  (FERM BP-6973) 
               
               
                 and  Enterococcus faecalis  (FERM BP-7231) killed cells on 
               
               
                 
                   Shigella flexneri 
                 
               
             
          
           
               
                   
                 
                   L. clearans 
                 
                 
                   Shigella flexneri 
                 
               
             
          
           
               
                 Number 
                 FERM BP-6973 
                 S type 
                 R type 
                 Ratio 
               
               
                 subculture 
                 cells/mL 
                 cells/mL 
                 cells/mL 
                 of R type 
               
               
                   
               
             
          
           
               
                 1 
                 1.2 × 10 9   
                 3.5 × 10 9   
                 0 
                 0 
               
               
                 3 
                   1 × 10 9   
                   3 × 10 9   
                 0 
                 0 
               
               
                 5 
                   7 × 10 8   
                 2.5 × 10 9   
                   2 × 10 8   
                 7.4%  
               
               
                 7 
                   6 × 10 8   
                   2 × 10 9   
                   3 × 10 8   
                 13% 
               
               
                 9 
                  7 × 10 8   
                   2 × 10 9   
                   1 × 10 9   
                 33% 
               
               
                 11 
                   5 × 10 8   
                   2 × 10 9   
                   1 × 10 9   
                 33% 
               
               
                 13 
                   5 × 10 8   
                 1.8 × 10 9   
                 1.2 × 10 9   
                 40% 
               
               
                 15 
                   4 × 10 8   
                 1.8 × 10 9   
                 1.5 × 10 9   
                 45% 
               
               
                 20 
                   5 × 10 8   
                 1.5 × 10 9   
                 1.8 × 10 9   
                 55% 
               
               
                 25 
                   6 × 10 8   
                   1 × 10 9   
                 1.5 × 10 9   
                 60% 
               
               
                 30 
                 4.5 × 10 8   
                  8 × 10 8   
                 1.8 × 10 9   
                 69% 
               
               
                 35 
                   4 × 10 8   
                   3 × 10 8   
                   2 × 10 9   
                 87% 
               
               
                 40 
                 3.5 × 10 8   
                   1 × 10 8   
                   2 × 10 9   
                 95% 
               
               
                 45 
                   4 × 10 8   
                 0 
                   2 × 10 9   
                 100%  
               
               
                 50 
                   4 × 10 8   
                 0 
                   2 × 10 9   
                 100%  
               
               
                   
               
             
          
         
       
     
     Example 6 
     A viable cell preparation of  Lactobacillus clearans  FERM BP-6972 or  Lactobacillus clearans  FERM BP-6972 prepared by the method in Manufacturing Example 1, a viable cell preparation of  Enterococcus faecalis  FERM BP-7230 or  Enteroccus faecalis  7231 prepared by the method in Manufacturing Example 2, and a killed cell preparation of  Enterococcus faecalis  FERM BP-7230 or  Enterococcus faecalis  7231 prepared by the method in Manufacturing Example 3 were combined and added to feed, which was given to groups of five 10-week old male mice. 1 g feed contained 5×10 8  cells  Lactobacillus clearans  and 5×10 8  cells  Enterococcus faecalis  (2.5×10 8  cells each of viable and killed cells when used in combination). The animals were allowed to feed freely for 3 months. The triglycerides and cholesterol in serum were measured for comparison with levels in control mice fed only normal feed. The triglycerides were measured with a Triglyceride E-Test Wako, while the cholesterol was measured with a Triglyceride C-Test Wako. The mean levels for each group were obtained. The mean level for the mice in the control group was 100%. The results are given in Table 35. Tables 35 and 21 show that the addition of viable or killed, as well as viable and killed, cells of  Enterococcus faecalis , which have action in lowering triglycerides and cholesterol, to  Lactobacillus clearans  was far more effective than when  Enterococcus faecalis  was administered alone. Unlike the results obtained when administered alone, the combined use of killed cells resulted in greater effects than viable cells. Compared to the control group, levels decreased as much as about ½. 
     
       
         
               
             
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 35 
               
             
             
               
                   
               
               
                 effect of mixture of  Lactobacillus clearans and   
               
               
                   Enterococcus faecalis  on triglycerides and cholesterol in 
               
               
                 mouse serum 
               
             
          
           
               
                   
                 
                   Enterococcus 
                 
                   
               
               
                   
                 
                   faecalis 
                 
               
               
                 
                   Lactobacillus 
                 
                 FERM BP-No. 
               
             
          
           
               
                 
                   clearans 
                 
                 Viable 
                 Killed 
                   
                   
               
               
                 FERM BP-No. 
                 cells 
                 cells 
                 Triglycerides 
                 Cholesterol 
               
               
                   
               
               
                 6972 
                 7230 
                   
                 60% 
                 55% 
               
               
                 6972 
                   
                 7230 
                 55% 
                 47% 
               
               
                 6972 
                 7230 
                 7230 
                 58% 
                 52% 
               
               
                 6971 
                 7231 
                   
                 57% 
                 55% 
               
               
                 6971 
                   
                 7231 
                 50% 
                 45% 
               
               
                 6971 
                 7231 
                 7231 
                 52% 
                 50% 
               
               
                   
               
             
          
         
       
     
     Example 7 
     7 groups comprising a total of thirty five 8-week old spontaneous hypertensive rats (SHR) were raised for 7 months on feed containing the cell preparations of Example 6. The blood pressure was measured before and after administration to groups given each type of cell preparation and the control group, with the results given in Table 36. Tables 36 and 22 reveal that the blood pressure had improved, on average, about 15% after 3 months. The killed cells of  Enterococcus faecalis  had more effective action than the viable cells, in the same manner as in Example 6. The drop in blood pressure was about 10 to 20%, results which were better than those obtained when  Enterococcus faecalis  was administered by itself. 
     
       
         
               
             
               
               
               
               
             
               
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 36 
               
             
             
               
                   
               
               
                 effect of mixture of  Lactobacillus clearans  and 
               
               
                   Enterococcus faecalis  on rat blood pressure 
               
             
          
           
               
                   
                 
                   Enterococcus 
                 
                 Mean 
                   
               
               
                   
                 
                   faecalis 
                 
                 level 
               
             
          
           
               
                 
                   Lactobacillus 
                 
                 FERM BP-No. 
                 before 
                 Mean level 
               
             
          
           
               
                 
                   clearans 
                 
                 Viable 
                 Killed 
                 admin. 
                 after admin. (mmHg) 
               
             
          
           
               
                 FERM BP-No. 
                 cells 
                 cells 
                 (mmHg) 
                   
                 Decrease 
               
               
                   
               
               
                 6971 
                 7230 
                   
                 210 
                 180 
                 14.3% 
               
               
                 6971 
                   
                 7230 
                 203 
                 168 
                 17.2% 
               
               
                 6971 
                 7230 
                 7230 
                 200 
                 175 
                 12.5% 
               
               
                 6973 
                 7231 
                   
                 207 
                 176 
                 15.0% 
               
               
                 6973 
                   
                 7231 
                 198 
                 162 
                 18.2% 
               
               
                 6973 
                 7231 
                 7231 
                 205 
                 178 
                 13.1% 
               
               
                 Control 
                   
                   
                 202 
                 210 
                 3.9% increase 
               
               
                   
               
             
          
         
       
     
     Table 37 shows the action in terms of various functions with the use of  Lactobacillus clearans, Enterococcus faecalis , and both. 
     
       
         
               
             
               
               
               
               
             
           
               
                 TABLE 37 
               
             
             
               
                   
               
               
                 Comparative summary 
               
             
          
           
               
                   
                   
                   
                 
                   L. clearans 
                 
               
               
                   
                 
                   Lactobacillus 
                 
                 
                   Enterococcus 
                 
                 and 
               
               
                 Parameter 
                 
                   clearans 
                 
                 
                   faecalis 
                 
                 
                   E. faecalis 
                 
               
               
                   
               
               
                 Decrease of toxic, 
                 ◯ to □ 
                 Δ 
                 ⊚ 
               
               
                 malodorous intestinal 
               
               
                 putrefying substances 
               
               
                 Feces deodorization 
                 □ 
                 Δ 
                 ⊚ 
               
               
                 Growth of beneficial 
                 ◯ to □ 
                 Δ 
                 ⊚ 
               
               
                 enteric bacteria 
               
               
                 Suppression of harmful 
                 ◯ 
                 Δ 
                 ⊚ 
               
               
                 enteric bacteria 
               
               
                 Antiflatulent action 
                 ◯ 
                 Δ 
                 ⊚ 
               
               
                 Suppression of 
                 Δ 
                 Δ 
                 ⊚ 
               
               
                 pathogen growth 
               
               
                 Suppression of 
                 ◯ 
                 x 
                 ⊚ 
               
               
                 pathogen toxicity 
               
               
                 Triglyceride reduction 
                 Δ 
                 ◯ to □ 
                 ⊚ 
               
               
                 Cholesterol reduction 
                 Δ 
                 ◯ to □ 
                 ⊚ 
               
               
                 Hypotensive action 
                 Δ 
                 □ 
                 ◯ to ⊚ 
               
               
                 Overall evaluation 
                 Perceptible 
                 Imperceptible, 
                 Rapid, 
               
               
                 in preserving health, 
                 results with 
                 but results 
                 perceptible, 
               
               
                 stimulating recovery, 
                 moderate use 
                 gradually 
                 highly 
               
               
                 preventing aging 
                   
                 appear with 
                 effective 
               
               
                   
                   
                 moderate to 
                 results 
               
               
                   
                   
                 prolonged use 
               
               
                   
               
               
                 *: excellent; ◯: good; □: ordinary; Δ: weak; x: none  
               
             
          
         
       
     
     Health care and related approaches have gradually changed in our increasingly aging society. Our time, where the treatment of acute diseases and chronic diseases has been regarded with the utmost importance, is witnessing a shift, now or in the near future, to “preventive medicine”, and even “nutritional therapy,” in light of our older society. These are not treatments which are undertaken after illness sets in. Rather, approaches for creating health on one&#39;s own through a more educated intake of nutrition are now increasingly in the mainstream in order to naturally prevent illness. It is not overstating the case to suggest that the prevention of disease to avoid the need for treatment at all is in the vanguard of treatment in the 21st century. 
     The oral administration of the lactic acid bacteria preparation of the present invention gradually extends the influence of beneficial intestinal bacteria, steadfastly guards the intestinal mucosa, produces vitamins, and synthesizes amino acids, all the while suppressing the growth of foreign bacteria and foreign pathogens, diminishing their toxicity, and activating immunological functions under the guidance of the lactic acid bacteria of the preparation. Harmful intestinal bacteria are thus markedly reduced, and the production of malodorous putrefying substances is suppressed, resulting in significantly deodorized feces. As if under attack, the lactic acid bacteria of the preparation actively feeds on the intestinal putrefying substances, resulting in a cleaner intestinal environment and the normalization of the intestinal mucosa and surrounding vessels and nerves. 
     Purification of the intestines, which are the wellspring of human vitality, energy, blood and flesh, results in better absorption of vital substances and conversely in fewer toxic substances absorbed through the intestines. This inevitably results in better hepatic function, and thus in lower triglycerides and cholesterol in serum, as well as cleaner blood. Such revitalized blood results in lower blood pressure, so that vital substances such as hormones, enzymes, antibodies, and immunological substances are not prevented from being distributed throughout the body&#39;s entire system. Metabolism is thus improved, and all systemic functions are invigorated. That is, the polluted intestines are cleaned and allowed to return to their pristine condition, resulting in an overall improvement and a vital state of health with no hint of illness. It can be said that Metchnikoff&#39;s doctrine on longevity is now being realized a century later. 
     The lactic acid bacteria preparation of the present invention can be stored without any loss of titer for 2 to 3 years, allowing it to be produced in the form of portable goods which can be readily used anywhere at any time, and is thus of immeasurable value. In societies with populations of increasingly advanced age, more and more elderly individuals are likely to become bedridden; although the disposal of waste can become a problem, the deodorization of such waste would make such a task that much less disagreeable, and could be of service to caretakers.