Pharmaceutical preparation comprising a combination of streptococcus strains and lactobacillus strains
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example 1
In Vitro Test
Production of a Lactobacillus PBS Filtrate:
[0061]Lactobacillus LB21 was cultured in 5 ml MRS broth (37° C., 5% CO2, 8 h). 1% (450 μl) LB21 culture was reinoculated in 6 tubes containing 45 ml fresh pre-heated MRS broth and was incubated at 37° C. and 5% CO2 for 16 h. Thereafter the tubes were centrifuged at 3800 rpm at 20° C. for 20 min. The supernatants were discarded and the pellets were resuspended in 5 ml pre-heated PBS and incubated at 37° C. and 5% CO2 for 5 h. Once an hour the tubes were gently mixed. After the incubation, the tubes were centrifuged at 3800 rpm at 8° C. for 20 min. All the supernatants were collected in a new tube, and the pH was measured (4.0-4.3) The supernatants were then sterile filtered through a 0.22 μm filter. The filtrates were stored in −20° C. in aliquots of 1 ml in 10 ml tubes until use.
Production of Control PBS Filtrate:
[0062]25 ml PBS solution was adjusted to a pH value of 4.3. The solution was thereafter sterile filtered through a 0...
example 2a
)
In Vitro Test
[0066]A combination of different Lactobacillus strains and an α-Streptococci mix of α89a, α502, α505, α4, and α6 was further tested in view of the capacity to inhibit and kill MRSA strains using the PBS supernatant method. At start the cfu was 2×105 / ml of MRSA in the tubes. After 4, 6, 8, 10, 16 and 24 hours samples were taken. The results expressed in cfu / ml can be seen in the table below. The surprisingly better effect by the combination of Lactobacillus and α-Streptococci is due to a synergistic effect of the substances, e.g. specific acids and bakterocins, that are produced by the different strains. sup=supernatant
PBS-sup0 h4 h6 h8 h10 h16 h24 hLB212 × 1051 × 1056 × 1041 × 1045 × 1031 × 1020LB9312 × 1058 × 1043 × 1041 × 1048 × 1036 × 103LB451 × 1055 × 1041 × 1046 × 1032 × 1036 × 102LB669 × 1046 × 1043 × 1047 × 1035 × 1021 × 102LB991 × 1055 × 1041 × 1045 × 1031 × 1020LB871 × 1059 × 1046 × 1041 × 1041 × 1037 × 102LB641 × 1057 × 1041 × 1046 × 1031 × 1033 × 102LB21 + a...
example 2b
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In Vitro Test of Other Combinations
[0067]A further test was done according to Example 2a) but with new combinations of bacteria. Here LB931 and LB Ess-1 were tested in a combination with α89a and Streptococcus lactis L1A, and the concentration of remaining MRSA were measured after different time periods. The test strain was MRSA 22.
PBS-sup4 h6 h10 h12 h24 hLB Ess-11 × 1055 × 1042 × 1036 × 1020LB21 + 89a4 × 1033 × 101000LB931 + 89a3 × 1032 × 1024 × 10100LBEss-1 + 89a2 × 1036 × 101000LB21 + L1A5 × 1020000LB931 + L1A6 × 1029 × 101000LBEss-1 + L1A1 × 1024 × 101000
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Abstract
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