LACTIC ACID BACTERIUM, BLOOD IRON INCREASING AGENT, AND ANEMIA IMPROVING AGENT

Abstract

A lactic acid bacterium is provided which has high foodstuff suitability, easy to manufacture, and can increase the blood iron concentration, thereby improving anemia. A lactic acid bacterium of salt-tolerant that increases the blood iron concentration.

Claims

1. A lactic acid bacterium of salt-tolerant that increases the blood iron concentration.

2. The lactic acid bacterium according to claim 1, which is isolated in a brewing process of miso.

3. The lactic acid bacterium according to claim 1, wherein a proliferation rate is 30 times or more when cultured in a medium having a salinity of 12 w/v %.

4. The lactic acid bacterium according to claim 1, which is Tetragenococcus halophilus.

5. The lactic acid bacterium according to claim 1 4, which is a lactic acid bacterium of Accession number NITE BP-02318 or a lactic acid bacterium of Accession number NITE BP-03010.

6. A blood iron increasing agent containing the lactic acid bacterium according to claim 1.

7. An anemia improving agent containing the lactic acid bacterium according to claim 1.

8. The lactic acid bacterium according to claim 2, wherein a proliferation rate is 30 times or more when cultured in a medium having a salinity of 12 w/v %.

9. The lactic acid bacterium according to claim 2, which is Tetragenococcus halophilus.

10. The lactic acid bacterium according to claim 3, which is Tetragenococcus halophilus.

11. The lactic acid bacterium according to claim 2, which is a lactic acid bacterium of Accession number NITE BP-02318 or a lactic acid bacterium of Accession number NITE BP-03010.

12. The lactic acid bacterium according to claim 3, which is a lactic acid bacterium of Accession number NITE BP-02318 or a lactic acid bacterium of Accession number NITE BP-03010.

13. The lactic acid bacterium according to claim 4, which is a lactic acid bacterium of Accession number NITE BP-02318 or a lactic acid bacterium of Accession number NITE BP-03010.

14. A blood iron increasing agent containing the lactic acid bacterium according to claim 2.

15. A blood iron increasing agent containing the lactic acid bacterium according to claim 3.

16. A blood iron increasing agent containing the lactic acid bacterium according to claim 4.

17. A blood iron increasing agent containing the lactic acid bacterium according to claim 5.

18. An anemia improving agent containing the lactic acid bacterium according to claim

2.

19. An anemia improving agent containing the lactic acid bacterium according to claim 3.

20. An anemia improving agent containing the lactic acid bacterium according to claim 4.

Description

EXAMPLES

[0099] Hereinafter, the present invention will be specifically described based on Examples, but the present invention is not limited to these Examples.

Example 1

[0100] The measurements of the iron concentration of serum (the serum iron concentration) were performed 20 healthy Japanese over 20 years old and under 45 years old before and after (on the 14th and 28th days) ingestion of the lactic acid bacterium. Note that the serum iron concentration was measured using a colorimetric method.

[0101] As a method for ingesting the lactic acid bacterium, tablets (blood iron increasing agent, anemia improving agent) containing the lactic acid bacterium of the present invention (specifically, the lactic acid bacterium of Accession Number NITE BP-02318) and the trade name “Lactic acid bacterium Kurahana LTK-1” (manufactured by Ichibiki Co., Ltd.) were ingested orally together with water (10 tablets per day (containing 1250mg of “Lactic acid bacterium Kurahana LTK-1”). The ingestion period was 28 days. The results are shown in Table 1.

[0102] The safety of ingestion of the trade name “Lactic acid bacterium Kurahana LTK-1” was also confirmed, and adverse events related to the drug were not confirmed.

[0103] Ingredients of orally ingested tablets are as follows: The trade name “Lactic acid bacterium Kurahana LTK-1” was 41.7% by mass, dextrin was 39.0% by mass, powdered cellulose was 15.0% by mass, sucrose fatty acid ester was 2.0% by mass, fine particulate silicon dioxide was 2.0% by mass, and shellac was 0.3% by mass.

TABLE-US-00001 TABLE 1 Before ingestion Day 14 Day 28 Serum iron 92.9 ± 9.0 107.8 ± 9.7 119.6 ± 11.9 (μg/dL)

[0104] As is clear from Table 1, it can be seen that the serum iron concentration after ingestion (day 14 and day 28 from the start of ingestion) is significantly increased compared to before ingestion of the tablet (blood iron increasing agent, anemia improving agent) containing the lactic acid bacterium of the present invention.

[0105] Then, from this result, it can be seen that those containing lactic acid bacterium of the present invention (blood iron increasing agent, anemia improving agent) can satisfactorily increase the serum iron concentration. Therefore, it is found that it exerts an effect on the improvement of anemia (in particular, iron deficiency anemia) (it can be used as an anemia improving agent).

[0106] The above results were checked for significance. The significances were confirmed by Wilcoxon signed rank test. The risk rate (significance level) was 5%.

[0107] (Evaluation of Salt Tolerance of Lactic Acid Bacterium)

When the salinity of the medium was set at 12 w/v %, the proliferation rate was tested on the trade name “Lactic acid bacterium Kurahana LTK-1”, on the lactic acid bacterium of Accession Number NITE BP-03010 (hereinafter, sometimes referred to as “ICK-3 Strain” or “ICK-3”), on “ICK-4 Strain” (sometimes referred to as “ICK-4”), and on “ICK-5 Strain” (sometimes referred to as “ICK-5”). Specific description will be made below.

[0108] (Medium)

As a nitrogen source and a trace mineral content, soy sauce (trade name “Koikuchi Soy Sauce” manufactured by Ichibiki Co., Ltd.) was used. As a carbon source, glucose (manufactured by Kanto Chemical Co., Inc.) was used. As other raw materials, salt (manufactured by Kanto Chemical Co., Inc.) and water were used. In this way, a medium consisting only of food raw materials was prepared.

[0109] Specifically, the above medium was prepared by mixing Koikuchi Soy Sauce, glucose, and salt with water so that Koikuchi Soy Sauce was 20 v/v %, glucose was 1.7 w/v %, and salinity was 12 w/v %, and then adjusting the pH to 7.0 with sodium hydroxide (Kanto Chemical Co., Inc.), a food additive.

[0110] The prepared medium was placed in a test tube (diameter: 18 mm×180 mm) in a volume of 10 mL, capped with SILICOSEN®, and sterilized in an autoclave at 121° C. for 15 minutes.

[0111] (Culture)

Lactic acid bacterium that had been pre-cultured in the above medium was added at a rate of 1 v/v %, assuming subculture. At this time, the number of bacteria at the time of initial culturing was 1.0×10.sup.7 cfu/mL. This was cultured statically for 20 hours in an incubator at 30° C.

[0112] (Measurement of Number of Viable Cell)

[0113] After the static culture, the number of viable cells was measured. Determination of number of viable cells was performed by applying diluted bacterial solution to “10SG10N plating medium”, followed by culturing (anaerobic culture at 30° C. for 4 days), and then counting the number of colonies.

[0114] The “10SG10N plating medium” containing 10 v/v % soy sauce (the trade name “Koikuchi Soy Sauce” manufactured by Ichibiki Co., Ltd.), 1.0 w/v % glucose, 1.0 w/v % yeast extract, 0.5 w/v % polypeptone, 0.2 w/v % sodium acetate 3 hydrate, 10 w/v % sodium chloride, 0.0025 w/v % “Tween80”, 0.02 w/v % magnesium sulfate 7 hydrate, 0.001 w/v % manganese sulfate 4 hydrate, 0.001 w/v % iron sulfate 7 hydrate was pH6.8 and 2 w/v % agar.

[0115] The value obtained by dividing the number of bacteria after 20 hours of culturing by the number of bacteria at the time of initial culturing (the number of bacteria after 20 hours/the number of bacteria at the time of initial culturing) was calculated as the proliferation rate (times/20 hours) for 20 hours. Table 2 shows the results.

TABLE-US-00002 TABLE 2 LTK-1 ICK-3 ICK-4 ICK-5 Initial Number of Lactic 1.0E+07 8.20E+06 5.20E+06 6.80E+06 acid bacterium Strains (cfu/mL) After Number of Lactic 3.5E+08 3.20E+08 5.90E+08 4.00E+08 20 acid bacterium hrs Strains (cfu/mL) Proliferation Rate of 34 39 113 59 Lactic acid bacterium (times/20 hrs)

[0116] It can be seen that all of the lactic acid bacterium of the present invention proliferates vigorously even at a salinity of 12 w/v %. On the other hand, under such conditions of high salinity, it is usually difficult for other bacteria (contaminants such as unwanted bacteria) to proliferate. Therefore, by culturing lactic acid bacterium under conditions of high salinity, the lactic acid bacterium of the present invention is preferentially cultured even if contaminants are mixed, so that a culture can be easily obtained.

Example 2

[0117] Blood tests were performed on 9 healthy long-distance athletes from a company's team aged 22-27 years before (2 months, 4 months, 6 months) and after (2 months, 4 months) ingestion of the lactic acid bacterium. The blood test was performed once every two months as described above.

[0118] Table 3 to Table 6 show the results of hemoglobin, serum iron, and ferritin in blood test items before and after ingestion of the lactic acid bacterium. In Tables 3 to 5, the “Mean value before the start of ingestion” indicates the mean value of each person of the blood tests performed 3 times before ingestion of the lactic acid bacterium (two months before, four months before, and six months before the start of ingestion). In addition, the “Mean value after the start of ingestion” indicates the mean value of each person of the blood tests performed twice after the ingestion of lactic acid bacterium (in the second month and the fourth month from the start of ingestion of the lactic acid bacterium).

[0119] In Table 6, the “Mean value before the start of ingestion” indicates the mean value of each person (A-I) of the blood tests performed 3 times before ingestion of the lactic acid bacterium (two months before, four months before, and six months before the start of ingestion). In Table 6, “2 months after the start of ingestion” indicates the results of each person (A-I) of the blood test performed two months after the start of ingestion of the lactic acid bacterium. “Four months after the start of ingestion” indicates the results of each person (A-I) on the blood test performed 4 months after the start of ingestion of the lactic acid bacterium.

[0120] As a method for ingesting the lactic acid bacterium, tablets (blood iron increasing agent, anemia improving agent) containing the lactic acid bacterium of the present invention (specifically, the lactic acid bacterium of Accession Number NITE BP-02318) and the trade name “Lactic acid bacterium Kurahana LTK-1” (manufactured by Ichibiki Co., Ltd.) were ingested orally together with water (2 tablets per day (containing 250mg of “Lactic acid bacterium Kurahana LTK-1”). The ingestion period was 4 months as described above. The diet during the test was a normal diet that was not changed from usual diet.

[0121] Ingredients of orally ingested tablets are as follows: The trade name

[0122] “Lactic acid bacterium Kurahana LTK-1” was 41.7% by mass, dextrin was 39.0% by mass, powdered cellulose was 15.0% by mass, sucrose fatty acid ester was 2.0% by mass, fine particulate silicon dioxide was 2.0% by mass, and shellac was 0.3% by mass.

[0123] The results of the blood tests are shown in Tables 3-6 below. Table 3 shows the change in serum iron concentration. Table 4 shows the change in ferritin concentration. Table 5 and Table 6 show the changes in hemoglobin concentration. “Serum iron concentration” is a value measured by a colorimetric method. “Ferritin concentration” is a value measured by CLIA (chemiluminescent immunoassay). “Hemoglobin concentration” is a value measured by SLS-hemoglobin method.

TABLE-US-00003 TABLE 3 Mean value before Mean value after the start of ingestion the start of ingestion Serum iron (μg/dL) 107 ± 30.1 122 ± 31.8 P = 0.07

[0124] As is apparent from Table 3, the serum iron concentration after ingestion of the tablet (blood iron increasing agent, anemia improving agent) containing the lactic acid bacterium of the present invention tends to increase as compared with before ingestion of the lactic acid bacterium of the present invention.

TABLE-US-00004 TABLE 4 Mean value before Mean value after the start of ingestion the start of ingestion Ferritin (ng/mL) 39.6 ± 20.6 54.2 ± 32.0 P = 0.03

[0125] As is apparent from Table 4, the concentration of ferritin after ingestion of the tablet (blood iron increasing agent, anemia improving agent) containing the lactic acid bacterium of the present invention is significantly increased as compared with before ingestion of the lactic acid bacterium of the present invention.

TABLE-US-00005 TABLE 5 Mean value before the Mean value after start of ingestion the start of ingestion Hemoglobin (g/dL) 14.2 ± 0.6 14.5 ± 0.7 P = 0.06

[0126] As is apparent from Table 5, the hemoglobin concentration after ingestion of the tablet (blood iron increasing agent, anemia improving agent) containing lactic acid bacterium of the present invention tends to increase as compared with before ingestion of the lactic acid bacterium of the present invention.

[0127] Table 6 shows the results of the blood tests performed on each tested person A to I, and it can be seen that the hemoglobin concentration tended to increase in 8 out of 9 tested persons in the blood test performed 4 months after the start of the ingestion of the lactic acid bacterium. In particular, it is considered that the tendency appears strongly in the tested person B, G, and I.

TABLE-US-00006 TABLE 6 Mean value before 2 months after the 4 months after the the start of ingestion start of ingestion start of ingestion (g/dL) (g/dL) (g/dL) A 14.0 14.4 15.0 B 14.8 13.4 15.5 C 13.7 13.1 14.4 D 14.5 13.4 14.2 E 13.7 14.5 14.2 F 14.1 14.6 15.1 G 14.4 14.3 16.1 H 14.0 14.4 14.3 I 14.4 14.4 15.5

Example 3

[0128] The test in this example was conducted in a double-blind, parallel-group study. Fourteen Japanese women over 20 years old and under 59 years old who were judged to have “mild anemia” with a hemoglobin level of less than 12 g/dL and serum ferritin level of less than 12 ng/mL were included as the tested person (the test subject). These 14 patients were assigned to two groups (group A and group B). Thereafter, the test subjects in Group A (the test food ingestion group) were made to ingest orally two tablets of the test food (containing the lactic acid bacterium of the present invention) per a day together with water every day, and the test subjects in Group B (the control food ingestion group) were made to ingest orally two tablets of the tablet control food (containing no lactic acid bacterium of the present invention) per a day together with water every day. The ingestion period was 8 weeks.

[0129] Ingredient percentages of orally ingested tablets (test food and control foods) are as follows:

(Test Food)

[0130] The test food included a trade name “Lactic acid bacterium Kurahana LTK-1” (manufactured by Ichibiki Co., Ltd.) of 41.7% by mass, a dextrin of 39.0% by mass, a powdered cellulose of 15.0% by mass, a sucrose fatty acid ester of 2.0% by mass, a fine particle silicon dioxide of 2.0% by mass, and a shellac of 0.3% by mass.

(Control Food)

[0131] The control food included a dextrin of 79.7% by mass, powdered cellulose of 15.0% by mass, a sucrose fatty acid ester of 2.0% by mass, a fine particle silicon dioxide of 2.0% by mass, a caramel pigment of 1.0% by mass, and a shellac of 0.3% by mass.

[0132] In this example, the values of hemoglobin, serum iron, reticulocyte count, total iron binding capacity (TIBC), and serum ferritin, respectively, were measured before the start of ingestion and 8 weeks after the start of ingestion. The hemoglobin was measured by SLS-hemoglobin method, the reticulocyte count was measured by the flow cytometry method, the serum iron was measured by the colorimetric method, TIBC was measured by the colorimetric method, and the serum ferritin was measured by CLIA (chemiluminescent immunoassay). The results are shown in Table 7.

[0133] As can be seen in Table7, in the control food ingestion group, there was no change in hemoglobin, but serum iron decreased, TIBC increased and serum ferritin decreased. This suggests that in the control food ingestion group, iron deficiency may be compensated for by storage iron to stop the decrease in hemoglobin.

[0134] On the other hand, in the test food ingestion group, both serum iron and serum ferritin increased suggesting that iron supply from food may exceed consumption. In addition, it has been reported that when iron deficiency anemia was treated with an iron preparation, the reticulocyte count increased in several days and the hemoglobin normalized in the subsequent six to eight weeks (See, Okada Sadamu: Treatment of Iron Deficiency Anemia, The Journal of the Japanese Society of International Medicine 99, 1220-1225 (2010)). Based on these reports, hemoglobin was not changed in the test food ingestion group as well as in the control food ingestion group in the results of the study period (8 weeks) in this example, but the reticulocyte count was increased in the test food ingestion group, suggesting that hemoglobin is an increasing situation.

[0135] Therefore, in this example, the test period is 8 weeks, but if the test period is 8 weeks or longer, it is presumed that an increase in hemoglobin is confirmed.

TABLE-US-00007 TABLE 7 Control food ingested group Test food ingested group (mean value) (mean value) Before the Before the start of 8 weeks after the start of 8 weeks after the ingestion start of ingestion ingestion start of ingestion Blood Hemoglobin (g/dL) 10.80 ± 1.03  10.53 ± 0.91  10.8 ± 1.09 10.9 ± 1.47 test Reticulocyte count (%) 9.4 ± 2.9 9.7 ± 1.7 10.1 ± 2.3  11.9 ± 3.3  item Serum iron (μg/dL) 46.1 ± 20.2 41.9 ± 7.4  45.4 ± 20.9 49.3 ± 27.5 TIBC (μg/dL) 431.0 ± 34.8  457.4 ± 44.2  440.0 ± 55.0  443.6 ± 54.9  Serum ferritin (ng/mL) 6.79 ± 1.98 6.20 ± 1.81 5.29 ± 1.82 6.50 ± 2.87

[0136] In this example, the safety of ingestion of the trade name “Lactic acid bacterium Kurahana LTK-1” was also confirmed, and as a result, the adverse effect was not confirmed.

Example 4

[0137] Three-week-old female mice (C57BL/6JJcl) were divided into eight groups containing 5 mice, and anemia model mice were prepared by being ingested low iron feed (powder) for 3 weeks to be anemic condition to all mice in all groups. During the following two weeks, the animals were fed each of the following feeds. Tables 9 to 12 show the blending prescription of the low iron feed (powder) and the normal feed. Table 9 shows the blending content of the feed (low iron feed (powder), normal feed), Table 10 shows the blending content of the respective mineral Mix of the low iron feed (powder) and the normal feed, and Table 11 shows the blending content of the vitamin Mix in the feed. In addition, Table 12 shows the content of iron in the feed (low iron feed (powder), normal feed).

[0138] The group fed the low iron feed containing 0.2% by mass of the lactic acid bacterium of Accession Number NITE BP-02318 (trade name “Lactic acid bacterium Kurahana LTK-1” (manufactured by Ichibiki Co., Ltd.)) was designated as the “LTK-1 feed group”, the group fed the low iron feed containing 0.2% by mass of salt-tolerant lactic acid bacterium “ICK-3” was designated as the “ICK-3 feed group”, the group fed the low iron feed containing 0.2% by mass of salt-tolerant lactic acid bacterium “ICK-4” was designated as the “ICK-4 feed group”, and the group fed the low iron feed containing 0.2% by mass of salt-tolerant lactic acid bacterium “ICK-5” was designated as the “ICK-5 feed group”. Three strains (No. 185 Strain (sometimes referred to as “No. 185”), No. 259 Strain (sometimes referred to as “No. 259”) and No. 448 Strain (sometimes referred to as “No. 448”) were also evaluated. The group fed the low-iron feed containing 0.2% by weight of salt-tolerant lactic acid bacterium “No. 185” was designated as the “No. 185 feed group”, the group fed the low-iron feed containing 0.2% by weight of salt-tolerant lactic acid bacterium “No. 259” was designated as the “No. 259 feed group”, and the group fed the low-iron feed containing 0.2% by weight of salt-tolerant lactic acid bacterium “No. 448” was designated as the “No. 448 feed group”. Furthermore, the group fed the low iron feed containing no salt-tolerant lactic acid bacterium was designated as the “low iron feed group”. All of the lactic acid bacterium used in this example belongs to Tetragenococcus halophilus.

[0139] In addition, during the test, the group in which normal food (“basic feed CE-2” manufactured by CLEA Japan, Inc.) was ingested was designated as the “normal feed group”. This normal feed group is mice that are not fed a low iron feed (powder) and are not anemic condition.

[0140] The breeding environment of the mice in each group was the same, specifically, as follows. Temperature was 20-26° C., humidity was 45-70% (excluding short-term changes during disinfection, etc.), and ventilation frequency was 10-15 times/hour. Illumination times ranged from 7:00 to 19:00 for bright hours and from 9:00 to 7:00 for dark hours. The microbiological grade was Specific pathogen free animals (SPF) (for aseptic breeding of experimental animals), the breeding racks were vinyl isolators (Max 8 cages), and the breeding cages were mouse PC (182×260×128 mm).

[0141] Drinking water was filled into a water supply bottle (250 cc), and then water was supplied after high pressure steam sterilizing (121° C., 30 minutes). The timing of water supply was 1 bottle/2 times/week. The feeder used a special cover for cages, the bedding used shaving chips (high pressure steam sterilizing at 121° C. for 30 minutes) and the cage changed once a week.

[0142] Blood samples were collected at 3 weeks (i.e., at the time of feed switching) and 5 weeks (i.e., 2 weeks after feed switching) after the start of breeding, and blood hemoglobin concentrations were measured using a blood cell counter for animals (fully automatic blood cell counter) “Celltac α MEK-6458 (manufactured by Nihon Kohden). The results are shown in Table 8.

TABLE-US-00008 TABLE 8 Week 3 Week 5 (2 weeks (at feed switching) after feed switching) Normal feed group 13.66 ± 0.34 13.14 ± 0.87 LTK-1 feed group 10.56 ± 1.33 11.86 ± 0.55 ICK-3 feed group 10.58 ± 0.92 11.58 ± 0.83 ICK-4 feed group 10.53 ± 0.86 12.15 ± 0.98 ICK-5 feed group 10.60 ± 0.88 12.02 ± 0.90 No. 185 feed group 10.62 ± 0.95 10.82 ± 1.05 No. 259 feed group 10.52 ± 0.87 11.32 ± 1.28 No. 448 feed group 10.58 ± 1.15 10.89 ± 1.35 Low iron feed group 10.54 ± 0.90 10.50 ± 0.46 **LTK-1 feed group, ICK-3 feed group, ICK-4 feed group, and ICK-5 feed group are significantly different from the low iron feed group in the risk rate of less than 5%.

[0143] As can be seen from Table 8, the blood hemoglobin concentration was increased in all groups that ingested the salt-tolerant lactic acid bacterium compared to the low iron feed group. Especially, in LTK-1 feed group, ICK-3 feed group, ICK-4 feed group and ICK-5 feed group, the blood hemoglobin concentration increased significantly compared with the low iron feed group. From these results, it was confirmed that ingesting feeds containing salt-tolerant lactic acid bacterium such as LTK-1, ICK-3, ICK-4, ICK-5, No. 185, No. 259, and No. 448 were effective in improving anemia. In particular, it is considered that lactic acid bacterium of LTK-1, ICK-3, ICK-4, ICK-5 are highly effective in improving anemia.

TABLE-US-00009 TABLE 9 Content of feed (%) Cornstarch 61.000 Milk casein 22.000 Mineral Mix 7.000 Crystalline cellulose 5.000 Purified soybean oil 4.000 Vitamin Mix 1.000 Total 100.000

TABLE-US-00010 TABLE 10 Low iron feed (powder) Normal feed Mineral Mix (mg/100 g feed) (mg) (mg) KH.sub.2PO.sub.4 1,730.00 1,730.00 CaHPO.sub.4 2H.sub.2O 1,500.00 1,500.00 CaCO.sub.3 1,355.40 1,355.40 MgSO.sub.4•7H.sub.2O 800.00 800.00 Cornstarch 990.00 800.00 NaCl 600.00 600.00 FeC.sub.6H.sub.5O.sub.7•nH.sub.2O — 190.00 MnSO.sub.4•5H.sub.2O 15.40 15.40 2ZnCO.sub.3•3Zn(OH).sub.2•H.sub.2O 6.00 6.00 Ca(IO.sub.3).sub.2 1.54 1.54 CuSO.sub.4•5H.sub.2O 1.26 1.26 CoCl.sub.2•6H.sub.2O 0.40 0.40 Total 7,000.00 7,000.00

TABLE-US-00011 TABLE 11 Vitamin Mix (mg/100 g feed) Cornstarch 635.378 Choline chloride 300.000 Vitamin E (50%) 20.000 Inositol 15.000 PABA 10.150 Nicotinic acid 10.150 D- Pantothenic acid Ca 2.000 Vitamin B2 1.872 Vitamin B1 1.500 Vitamin A (1 million IU/g) 1.200 Vitamin B6 1.020 Biotin (2%) 0.500 Vitamin D3 (0.5 million IU/g). 0.480 Vitamin K3 0.300 Vitamin B12 (2%) 0.250 Folic acid 0.200 Total 1,000.000

TABLE-US-00012 TABLE 12 Fe content (round off the 6th decimal place) Low iron feed (powder) Normal feed Blending Blending Raw material Content ratio Content ratio Content name (%) (%) (%) (%) (%) Milk casein  0.0008 22.000000 0.00018 22.000000 0.00018 Cornstarch  0.0003 62.625378 0.00019 62.435378 0.00019 FeC.sub.6H.sub.5O.sub.7•nH.sub.2O 16.6700 — —  0.190000 0.03167 Total (%) — — 0.00036 — 0.03204

[0144] (Results and Discussion)

[0145] As shown in Table 1, it can be seen that the lactic acid bacterium of the present invention (the blood iron increasing agent of the present invention) has an effect of increasing the amount of iron in human serum by oral ingestion for a predetermined period of time. Further, from the results shown in Table 1, it can be seen that the lactic acid bacterium of the present invention (the blood iron increasing agent of the present invention) can be utilized as one capable of improving anemia (particularly, iron deficiency anemia).

[0146] From the results of Table 3 to Table 6, it can be seen that those containing the lactic acid bacterium of the present invention (blood iron increasing agent, anemia improving agent) can satisfactorily increase the serum iron concentration, the ferritin concentration, and the hemoglobin concentration. From this, it can be seen that it exerts an effect on the improvement of anemia (especially, iron deficiency anemia) (it can be used as an anemia improving agent).

[0147] Further, those containing the lactic acid bacterium of the present invention (blood iron increasing agent, anemia improving agent) exerts an effect on the improvement of sports anemia of athletes and people who are encouraging sports for health by increasing iron in blood (it can be used as an anemia improving agent). In addition to such an effect, an effect of improving the performance of the athlete (for example, an effect of improving the physical ability such as endurance) can be expected by enhancing aerobic exercise by promoting efficient oxygen transport in the body of the endurance athlete who is a player such as long-distance running.

[0148] Furthermore, from the results of Example 3 (Table 7), it was found that serum iron and serum ferritin were increased by ingesting the lactic acid bacterium of the present invention. Therefore, according to the lactic acid bacterium of the present invention, improvement of indefinite complaints due to latent anemia (latent iron deficiency) caused by lowering of serum ferritin can be expected.

[0149] In addition, from the results of Example 4 (Table 8), it can be seen that in addition to the lactic acid bacterium of Accession Number NITE BP-02318, it is also effective to increase the amount of iron in the serum by oral ingestion for a predetermined period of time for the lactic acid bacterium of Accession Number NITE BP-03010 (ICK-3 Strain), ICK-4 Strain, ICK-5 Strain, No. 185 Strain, No. 259 Strain, and No. 448 Strain. Although Example 4 is a mouse test, it can be inferred that the same effect is exhibited in humans.

INDUSTRIAL APPLICABILITY

[0150] The lactic acid bacterium of the present invention can be used as an active ingredient of a blood iron increasing agent or an anemia improving agent. The blood iron increasing agent of the present invention can be utilized as one for increasing the amount of iron in blood. The anemia improving agent of the present invention can be utilized as one for improving anemia (particularly, iron deficiency anemia).

Accession Number

[0151] (1) Depositary Institution: NITE Patent Microorganisms Depositary, National Institute of Technology and Evaluation

Depositary Institution Address: Room 122, 2-5-8 Kazusakamatari, Kisarazu-shi, Chiba 292-0818, Japan

[0152] Date of deposit: Sep. 6, 2017
Accession number NITE BP-02318

(2) Depositary Institution: NITE Patent Microorganisms Depositary, National Institute of Technology and Evaluation

Depositary Institution Address: Room 122, 2-5-8 Kazusakamatari, Kisarazu-shi, Chiba 292-0818, Japan

[0153] Date of deposit: Aug. 5, 2019
Accession number NITE BP-03010