GB1584693A – Bacterial deodorant composition
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GB1584693A – Bacterial deodorant composition
– Google Patents
Bacterial deodorant composition
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Publication number
GB1584693A
GB1584693A
GB21343/77A
GB2134377A
GB1584693A
GB 1584693 A
GB1584693 A
GB 1584693A
GB 21343/77 A
GB21343/77 A
GB 21343/77A
GB 2134377 A
GB2134377 A
GB 2134377A
GB 1584693 A
GB1584693 A
GB 1584693A
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Prior art keywords
composition
days
bacterial
bacteria
strain
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1976-05-21
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GB21343/77A
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SEIKEN KAI FOUNDATIONAL JURIDI
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SEIKEN KAI FOUNDATIONAL JURIDI
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1976-05-21
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1977-05-20
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1981-02-18
1977-05-20
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SEIKEN KAI FOUNDATIONAL JURIDI
1981-02-18
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A—HUMAN NECESSITIES
A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
A61L9/00—Disinfection, sterilisation or deodorisation of air
A61L9/01—Deodorant compositions
Description
(54) BACTERIAL DEODORANT COMPOSITION
(71) We, SEIKEN KAI FOUNDATIONAL JURIDICAL PERSON, a legal body organized under the laws of Japan, of 95, Fushimido-cho, Tondabayashi
City, Osaka Prefecture, Japan, do hereby declare the invention, for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following statement:- This invention relates to a deodorant composition, a bacterial preparation containing such composition, and to a method for the deodorization of excrement using such a comnosition or preparation.
According to one aspect of the present invention, there is provided a composition
comprising only one or a small number of strains of an autotrothic bacteria, said
bacterial strain(s), when cultivated at 280C exhibiting a ,u (specific rate of growth) value of 0.25 more than that exhibited by colon bacteria in an S-W medium when the composition of the culture medium (Stephenson-Whetham) consists of 1 g of
KH2PO4, 0.7 g of MgSo4 . 7H20, 1 g of NaCI, 4 g of (NH.t)2HPO4, 0.03 g of FeSO4 . 7H2O and 5 g of glucose per litre of the culture medium; said bacterial strain, or said small number of strains in combination exhibiting accelerated growth in said culture medium in the presence of (a) each of Na2S . 9H20, ammonia and a lower aliphatic carboxylic acid and/or (b) at least one of cystine, cysteine, and methionine.
According to another aspect of the present invention, there is provided a bacterial preparation comprising (1) a composition as defined in the last preceding paragraph, and (2) a composition comprising (A) at least one of S-, N- and C-compounds, and/ or (B) at least one S-containing amino acid.
According to a further aspect of the present invention, there is provided a method for the deodorization of excrement, comprising contacting excrement with a deodorizing effective amount of a composition as defined in the last preceding paragraph 1, or a preparation as defined in the last preceding paragraph.
Experiments in the deodorization of excrement using microorganisms have been previously conducted and some microorganisms have been sold for the purpose of deodorizing excrement. However, such microorganisms were not commercially successful and their sales were discontinued. The reason why sales of such microorganisms were discontinued is believed to be due to (1) their lack of effectiveness in deodorization, (2) the difficulties for cultivation of such microorganisms, (3) the problems in storing them in such a way as to prevent them from losing their ability to deodorize,
(4) the uncertainty that they were not pathogenic due to the lack of information on such microorganisms, and (5) their cost effectiveness.
Investigations by the inventor with a view to finding a suitable microorganism for deodorizing excrement have proved extremely time consuming and difficult because of the extremely complex and variable composition of excrement. The number of compounds contained in excrement can vary very widely depending upon the food eaten by the animal producing the excrement and upon the great variety of biochemical reactions which take place in the animal’s intestine. The inventor has found that the unpleasant smelling ingredients of excrement can be divided broadly into three classes, namely compounds containing sulphur (S-compounds), compounds containing nitrogen (N-compounds) and compounds containing carbon (C-compounds). Of course, there are a very large number of individual compounds in excrement which fall within each broad class. This provides almost insuperable problems when conducting experiments in order to obtain repeatability. The inventor has found that sodium sulphide can represent S-compounds, ammonia, indole or skatole can represent
N-compounds and a lower aliphatic carboxylic acid can represent C-compounds.
Accordingly, the inventor conducted exhaustive experiments to find a bacterium or a small number of strains of bacteria which were capable of assimilating or chemically modifying these to remove the odor thereof. Any bacterium or strains of bacteria which are capable of deodorizing such selected compounds can be expected to have a deodorizing effect upon excrement provided that such bacteria are also capable of rapidly growing in excrement. This provides additional problems as follows:- (1) Excrement generally is of low nutritional value and therefore it is difficult to
obtain a high rate of multiplication of the bacteria.
(2) There are many bacteria present in excrement and the bacteria chosen for
deodorization should be capable of feeding on any nutrients in the excrement in
spite of the existence of the other bacteria.
(3) Excrement contains gall which generally prevents the multiplication of bacteria.
(4) The external surface of excrement can be exposed to air and so the bacteria must
be capable of growing under aerobic conditions. However, anaerobic conditions
exist in the interior of a mass of excrement and so the bacteria should be capable
of multiplying under anaerobic conditions.
(6) The bacteria should be pH resistant because of the different pH conditions which
can be encountered in excrement.
(7) Excrement is generally a mixture of faeces and urine and so bacteria should have
a high activity even in a medium where a high concentration of salt exists.
(8) The bacteria employed should not be capable of causing harm to animals or
plants.
(9) The bacteria should be capable of resisting anti-bacterial agents and to spices
which are commonly used in food.
Thus, it will be appreciated that enormous problems are encountered when seeking a single bacterial strain which will proliferate in excrement and which is capable of deodorizing S-compounds, N-compounds and C-compounds. Hereinafter the action of bacteria on S-compounds will be referred to as «S-action», and the actions on N-compounds and C-compounds will be referred to as N-action and Caction, respectively.
The inventor has succeeded in isolating various bacterial strains from a vast number of bacterial strains. Some of the isolated bacterial strains have only an Saction or an N-action or a C-action, others have two out of the three actions and others have all three actions. All of the isolated strains are capable of proliferating in excrement.
In Table 1 below, the effectiveness upon excrement of four bacterial strains is shown. Three of the strains, namely FRI Nos.: 2546, 2545 and 2544, are only strong in respect of one of the S-, N-, and C-actions. However, another isolated strain, namely FRI No. 2823 was found to have S-, N-, and C-actions as will be seen from the Table. It will be appreciated that, where a bacterial strain is used which does not have all three actions, then it will be used together with another bacterial strain or strains having the action or actions lacking in the first-mentioned strain.
TABLE 1
Residual Odor (Deodorizing power test) after standing for each given period at 28 C Characteri stic in Action F.R.. No. 24 hr 48 hr 72 hr S-action is 2546 2 1′ 1 especially strong N-action 2545 3 3 3 C-action is 2544 3 3 2′ especially strong S+N-actions are 2546+2545 1′ 1′ 0′ especially strong S+C-actions are 2546+2544 1′ 1′ 0′ especially strong N+C-actions are 2545+2544 3 2′ 2 especially strong S+N+C-actions, 2546+2545+2544 1′ 1′ 0′ all together S+N+C-actions, 2823 0’#1 0′ 0#0′ all together Remarks: # In each test, 10 wt% of culture liquor was added to excrement based upon
the weight of the latter.
# F.R.I. No. means the number of acceptance of the Fermentation Research
Institute (Agency of Industrial Science & Technology), Japan.
# The meanings for the figures for Residual Odor are as follows: O … completely deodorized (the most effective).
0′ … the odor was virtually undetectable,
1 … at first, it was just possible to detect a faint odor but the odor
then became undetectable.
l’ … at first, a faint odor was definitely detected but soon after, the
odor became undetectable.
2 … a faint odor was definitely detected and this faint odor remained.
2′ … the residual odor was much weaker than the strong odor of the
control sample (equal to the test sample).
3 … the residual odor was weaker than the strong odor of the control
sample.
3′ … only a little weaker than the strong odor of the control sample.
4 … almost equal in strength to the odor of the control sample.
However, it should be pointed out that even if the detected
strengths of the odor in both samples were the same, it does
not follow that the amounts of odiferous substances in both
samples are the same
Other characteristics of the four typical bacterial strains listed in Table 1 are given in Tables 2 to 5 below. It is to be noted that FRI No. 2546 has a strong Saction, FRI No. 2544 has a strong C-action and FRI No. 2545 has a strong N-action whereas, as noted above, FRI No. 2823 has strong S-, N-, and C-actions. These four strains represent a suitable cross-section of a wide variety of bacterial strains which are useful in the present invention. Table 2 below shows the results of microscopic tests on the above-mentioned four bacterial strains and their biochemical behaviour. Table 3 shows the ability of the bacterial strains to decompose various
Saccharides. Table 4 shows the nutritional demand of the bacterial strains during multiplication and also shows the result of tests of accelerate their growth. Table 5 shows the results obtained by cultivation of the bacterial strains when various effective components contained in excrement are added to the various culture media as nutrient.
As will be understood from Table 1, the rates of growth of those bacteria are accelerated by the addition of such nutrients as S-, N-, and C-compounds into the basic culture medium if the medium contains poor nutrients. However, as the nutritional value of the basic culture medium increases, the addition of such S-, N-, and
C-compounds becomes less effective to accelerate the growth rate. In other words, when a basic culture medium having a medium nutritional value is used for cultivation, only a weak acceleration of growth is observed when the S-, N-, and C-compounds are added; and an even weaker acceleration of growth is obtained in a highly nutritional basic culture medium when such compounds are added. In fact, under these last conditions, it was impossible to measure the acceleration of growth with the naked eye although acceleration of growth was capable of measurement using a nephelometer.
In the accompanying drawings;
Fig. 1 is a graph in which the log of bacterial growth is plotted against time in the case where a highly nutritional basic culture medium is employed. As will be appreciated from an examination of Fig. 1, no acceleration can be observed even though S-, N-, and C-compounds are added for the purpose of accelerating growth.
Fig. 2 is a graph similar to Fig. 1 in the case where a basic culture medium of medium nutritional value is employed. A weak tendency for growth acceleration can be seen in certain cases when the S-, N- and C-compounds are added.
Fig. 3 is a graph similar to Figs. 1 and 2 showing the case where a basic culture medium of low nutritional value is employed. As can be seen, a considerable acceleration of growth is experienced depending upon the concentration of the S-, N-, and
C-compounds added.
In each of Figs. 1 to 3, curve 0 corresponds to a curve obtained when no S-, N-, and C-compounds were added, and curves 1, 2 and 3 were obtained when 0.1 of a gram, 1 g, and 2 g, respectively, of each of sodium sulphide, ammonia and acetic acid were added to 1 litre of the culture medium.
In the case of Fig. 1, the low nutritional culture medium employed was Stephenson Whetham medium containing 1 g of KHoPO4, 0.7 g of MgSO . 7H2O, 1 g of
NaCI, 4 g of i(NH4)2HPOJ, 0.03 g of FeSO4 . 7H20 and 5 g of glucose. In the case of Fig. 2, the culture medium employed was the above-mentioned Stephenson
Whetham medium with the addition of 1 g of casamino acid and 0.1 g of vitamins.
In the case of Fig. 3, the culture medium employed was the aforementioned Stephenson Whetham medium with the addition of 10 g of petone, 5 g of broth, 5 g of NaGl and 1 g of glucose.
The sodium sulphide was added as Na2S . 9H20, the ammonia was added as the 37% aqueous solution and the acetic acid was added as glacial acetic acid. The addition of these compounds was effected after 21 hours from the beginning of cultivation, i.e. when the bacterial growth entered the logarithmic phase. A nephelometer was utilised to obtain the results and the response to addition of the sodium sulphide, ammonia and acetic acid was accurately obtained by measurement of the relative turbidity with elapse of time.
TABLE 2
O s io d Om 1 8 1 I + o ; Ca Motility + + + , o o for facultative facultative facultative facultative cultivation anaerobic anaerobic anaerobic anaerobic Oo 0 0 t of colony | + high upheaval low c z Japanese gelatin) upheaval upheaval wet smooth wet o»a wet a smooth surface Eo surface surface opaline wet opaline O opaline wet z wet O C’ N t DR X r = =e d- | ; l l S D u j E v = z evX Y t o , D o t O > 8 we a ot UO!}PA!IInO Jo sllnsaX pue UO!}PAI2SqO 3!d03S0J3!W .
TABLE 2 (Continued)
F.R.I. No. 2546 2545 2544 2823 Generation of NH3 Generation of H2S > ‘I I I I I of + I I I Co t t o Generation of coloring matter o Decomposition of urea + + + Utilization of citric + + t I I I l I + + I I I t-r of + gelatin V-P reaction Reduction of nitrate . e) ce z o Y : o o v c E c o < -, = = wo c W 9,, C t4 t C W C C , O O D Y sa!padoJd leo!ltia4oo!^d Remark: + ... positive, - ... negative TABLE 3
F.R.I. No. 2546 2545 2544 2823 Glucose + + + + Starch -. - - + Melezitose - - - Maltose Raffinose - - - - - Fructose + -- - -.
0 Melibiose - - - + o Xylose -- ± + + Sorbitol - - - -. Mannitol - - - O, Inositol -- -- - Arabinose - -- + Lactose -- - - Mannose - + - + Saccharose - - - Salicin ~ ~ ~ TABLE 4
F.R.I. No.
2546 2545 2544 2823 Substance added to (S-W) culture medium none ++ ++ ++ ++ ++ S-containing amino-acid +++ ++ ++ Cyclic amino-acid -+ ++ ++ ++ Amino-acid of chain structure ++ ++ ++ ++ Cystine +++ ++ ++ +++ Cysteine +++ ++ ++ Methionine +++ ++ ++ +++ Casamino-acid +++ +++ +++ +++ Casamino-acid + Vitamins +++ +++ +++ Casamino-acid + Extract of yeast +++ +++ +++ +++ Extract of yeast +++ +++ +++ Vitamins ++ TABLE 5
F.R.I. No. 2546 2545 2544 2543 Substance added to basic culture medium a b c d a b c d a b cd a b c d none - ++ +++ +++ - + ++ +++ - ++ ++ +++ - ++ ++ +++ Acetic acid - ++ +++ +++ - + ++ +++ - ++ +++ +++ + ++ +++ +++ Butyric acid + ++ +++ +++ - + ++ +++ - ++ +++ +++ + ++ +++ +++ Propionic acid + ++ +++ +++ - + ++ +++ - ++ +++ +++ + ++ +++ +++ Na2S.9H2O + ++ +++ +++ - + ++ +++ - ++ ++ +++ + ++ +++ +++ Mercaptan - ++ +++ +++ - + ++ +++ - ++ ++ +++ + ++ +++ +++ Ammonia - ++ +++ +++ + ++ +++ +++ - ++ ++ +++ + ++ +++ +++ Skatole - ++ +++ +++ + ++ +++ +++ - ++ ++ +++ + ++ +++ +++ Extract of Excrement ++ ++ +++ +++ ++ ++ +++ +++ ++ ++ +++ +++ ++ ++ +++ +++ Remarks: # The compositions of basic culture media were as follows: a ... [S-W] - glucose, b ... [S-W], c ... 8 g of peptone + 2 g of glucose, d ... 10 g of peptone + 5 g of broth + 5 g of NaCl + 1 g of glucose, such amounts being per litre of the basic culture medium.
# Those results shown in Table 4 and Table 5 were obained by observation with the naked eye.
Next, in Table 6, the experimental data on the specific growth rate (,,a) of two strains of bacteria when various kinds of basic culture media were used will be shown, wherein as one of them, a deodorization bacterium of the present invention, F.R.I.
No. 2823 was chosen as representative of the deodorization bacteria of the present invention and as an example of bacterium outside the scope of the present invention, a colon bacillus was used as a comparison. According to the value of the specific growth rate Ua, from many experiments, it was found that the value should preferably be more than 0.65 in order to have a high deodorization capability when (S-W) is used for the basic culture medium.
TABLE 6
value F.R.I. Colon Basic culture medium No. 2823 bacillus (s-w) - Glucose + F component 0.53 (s-w) 0.76 0.37 (S-W) + Amino-acid containing sulphur 0.82 0.41 (S-W) + F component 0.82 0.41 (S-W) + Amino acid + Vitamins 0.85 0.46 "E component" means a component containing S-, N- and C- compounds.
The deodorization bacteria should preferably have a high rate of growth as mentioned above, besides, it is necessary for the deodorization bacteria to be resistant to the acid component of gall contained in excrement, to various anti-bacterial agents, such as, for example, antibiotics, and also to spices commonly used in foods. Furthermore, it is desirable that the deodorization bacteria have an ability to resist chemical reagents which have a deodorization ability in order to make it possible to use the deodorization bacteria together with those chemical reagents.
In Table 7, there is demonstrated the effect on deodorization of the sensitivity of the bacteria to gall.
TABLE 7.
Change of residual odor with elapse of time (F.R.I. No. 2823) Sensitivity of bacteria to gall 24 hrs 48 hrs 72 hrs not sensitive 0' cm 1 0' O ~ 0' sensitive 3'. 3' ~ 3 3 One of the problems associated with deodorization using microorganisms is to ensure that the bacteria retains its deodorizing power for a long time. In the case of previously proposed bacteria having a high ability for deodorization, the ability of the bacteria to deodorize rapidly decreases with elapse of time even though they may be stored very carefully.
Additionally, it is very difficult to restore the deodorization capability of bacteria if this capability has been lost by cultivating the bacteria in the wrong medium or by carrying out repeated sub-culture. Thus, effective deodorization results cannot be obtained unless the bacteria can be cultured in such a way as to prevent the deteriora tion of the deodorization properties thereof. In other words, even if a bacterial deodorizing agent is originally extremely effective, it has no commercial advantage over chemical deodorizing agents if its deodorizing ability cannot be maintained.
Thus, it is extremely important to develop a suitable culture medium which does not spoil the deodorizing power of bacteria having good deodorization properties.
Moreover, it is important to develop culture media in which the required bacteria can be multiplied effectively.
The culture medium used in the deodorant composition of the present application possesses the above properties. The invention has succeeded in isolating a very large number of deodorizing bacteria of which 5 strains (i.e. FRI Nos.: 2823, 3577, 3576, 3575 and 3578) are typical examples. The following experimental results have been obtained using these 5 strains which have a strong deodorization ability and which each have S-, N- and C-actions. Since the deodorization abilities of these 5
strains were similar, the experimental results given below relate to the average results from the data obtained from the 5 strains. In the experiments, the culture medium employed contained 1% of gall powder. The data obtained in Table 8 below was
obtained by adding each culture medium to excrement in an amount of 10% by
weight and deodorization was effected for 24 hours at 28"C. In Table 8(A) below,
there are shown the deodorizing effects of those bacteria which were obtained by sub
culture using conventional culture media of high to low nutritional value. As is clearly
shown in Table 8(A), a rapid decrease in the deodorization activity of bacteria can
never be prevented if sub-culture is effected using conventional culture media.
TABLE 8-a
Residual odor generation Culture medium for sub-culture 1st 3rd 5th 7th (A) Broth + Pepton + Vitamins 2' 3 3'. 4 (B) Broth + Pepton 2' 3 3' 4 (C) Pepton ±Vitamins 2' 3 3' 4 (D) Pepton 2~ 2' 3 3 3' 4 (E) Amino-acid + Vitamins 22' 2' 3 3' 31N 4 (F) Amino-acid 2 2' 3 3' (G) (S-w) + Vitamins 1 1# 1' 2 3' (H) (S-W) 1' 2 2' 3' :In Table 8-b below, the tendency of the decrease of activity is shown, wherein the sub-culture media whose compositions were respectively (A), (B), (C)...and (H) shown in Table 8-a, together with 1 g/l of an F component, were used for subculture. The CCF component" is a component containing foul-smelling S-, N- and Ccompounds which are contained in excrement and, practically, in the experimental tests shown in Table 8-b, Na2S . 9H2O, NH3 and acetic acid, respectively, were used as representatives of those S-, N- and C-compounds.
TABLE 8-b
Residual odor generation Culture medium for sub-culture 1st 3rd 5th 7th (A) + F component 2 2' 3 3' (B) + F ,, 2 2' 3 3' (C) + F ,, 1' # 2 2 2' 3 (D) + F ,, 1' N 2 2 2' 3 (E) + F ,, 1' # 2 2 2' 2'#3 (F) + F ,, 1'# 2 2 2' 3 (G) + F ,, 1 1#1' 1' 1' (H) + F " 1 1' 1'~ 2 2 As will be clearly understood from Table 8-a and Table 8-b, the compositions (G), (H), (G) +F and (H) +F component came only a small reduction in deodorization ability.
With such compositions, further experiments were conducted in which the glucose component was omitted from the (S-W) medium and the results are shown in the following table, wherein - glucose means the glucose component was eliminated.
Residual odor generation Culture medium for sub-culture 1st 3rd 5th 7th TABLE 9-a
Residual odor Amino-acid added to basic culture generation medium of (G)-glucose for sub-culture 4th 11th Cystine 0'#1 1 Cysteine 1 1' Methionine 0's 1 1 Glycine 1 2 Glutamic acid 1 2 Arginine 2 2' Asparagine 2 2' Aspartic acid 2 Alanine 1' 2 Phenyl alanine 2 2' Amino-butyric acid 1' 2' Leucine 2' 3 Isoleucine 2' 3 Proline 2 2' Lysine 1' 2 Tyrosine 1' 2' Histidine 1 2' Tryptophan 2 3 Threonine 2 3 Serine 2 3 Casamino-acid 1'#2 2#2' TABLE 9-b
Residual odor Amino-acid added to basic culture generation medium of (H) -glucose for structure 4th 11th Cystine 1#1' 1'#2 Cystein 1#1'@ 2 Methionine 1#1' 2 Glycine 1' 2 Glutamic acid 1' 2' Arginine 2#2' 3 Asparagine 2' 3 Aspartic acid 2' 3 Alanine 2 2' Phenylalanine 2' 3 Amino-butyric acid @2 2'~ 3 Leucine 3 3' Isoleucine 3 3' Proline 2' . 3 Lysine 1' 2' Tyrosine 1' 2 2' Histidine 1#1' 2' Tryptophan 2#2' 3#3' Threonine 2#2' 3#3' Serine 2N 2' 3# 3 Casamino-acid 2 2' In the following Table 10-a and Table 10-b, the results of two series of experiments are summarized, wherein the activities of bacteria multiplied successively in repeated subculture using culture media of (G)-glucose or (H)-glucose to which various amino-acids and F component had been added, were examined.
TABLE 10-a
Residual odor Substance added to basic culture medium of generation (G)-glucose for sub-culture 4th 11th Cystine 0'#1 0'#1 Cystine 0'#1 0'#1 Methionine 0'#1 0'#1 Glycine 0'# 1 1 Glutamic acid 0'# 1 1 Arginine 1 1' F Asparagine 1 1' component Aspartic acid 1 1' Alanine 0'#1 1#1' + Phenyl alanine 1 1' Amino-butyric acid 0'# 1 1' Leucine 1 1'#2 Isoleucine 1 1'#2 Proline 1 1' Lysine 0'~ 1 1 Tyrosine 00N 1 1' Histidine 0'#1 1#1' Tryptophan 1 1'#2 Threonine 1 1'#2 Serine 1 1'#2 Casamino-acid 0' #1 1 ~ 1' TABLE 10-b
Residual odor Substance generation added to basic culture medium of (H)-glucose for sub-culture 4th 11th Cystine 1 1' Cysteine 1 1' Methionine 1 1' Glycine 1 1' Glutamic acid 1 1' Arginine 1 # 1' llN 2 F Asparagine 1' 2 component Aspartic acid 1' 2 Alanine 1 # 1' 2 + Phenyl alanine 1' 2 Amino butyric acid 1 N 1' 2 Leucine 1' 2 Isoleucine 1' 2 Proline 1' 2 Lysine 1 1' Tyrosine 1 N 1' 1' 1' Histidine 1 1' - 2 Tryptophan 1' 2 Threonine 1' 2 Serine 1' 2 Casamino-acid 11' l(y2 As will be clearly understood from Tables 9 and 10, amino-acids which are effective to keep the deodorizing power of the bacteria almost constant continuously for many generatiòns are cystine, cysteine, methionine, glycine, glutamic acid, arginine, amino-butyric acid, lysine, histidine, alanine, tyrosine. Other amino-acids used in those experiments cannot prevent a gradual decrease of the deodorizing power of the bacteria.
Further experiments on the bacteria cultivated by successive sub-culture proved that addition of starch, mineral and/or vitamin, has a remarkably good effect on the preservation of the deodorizing power. Furthermore, it is more beneficial to add any combination of starch, mineral, vitamin and effective amino acid (eg cystine) to the said culture media for the preservation of deodorizing power and such an addition was also effective to increase the rate of growth. These effects mentioned above will be clearly understood from Tables 11, 12 and 13.
TABLE 11
Residual smelling degree Substance generation added to (G)-glucose + F component 10th 15th 20th 25th Starch 0'#1 0'#1 1 Mineral O ' 1 1 1 1#1' Starch + Mineral 0'#1 0'#1 0'#1 0'# TABLE 12
Number of bacteria Substance (aerobic) Residual odor added to basic culture cultured for degree medium* 72 hrs at 280C (15th generation) none 5 x 10' 2' Starch 7x109 1'#2 Mineral 6 x 109 2 Vitamins 7x109 1' Starch + Mineral 12 x 109 1 ~ 1' Starch + Vitamins 13 x 109 1 Mineral + Vitamins 8 x 109 1 AT 1' Starch + Mineral + Vitamins 15 x 109 0'# 1 *Remark: The basic culture medium was (S-W) - glucose + F component.
TABLE 13
Residual odor Substance generation added to basic culture medium [(G)-glucose + F component] 20th 25th 30th 35th Starch + Mineral 0'#1 0'#1 1 Starch + Mineral + Cystine 0' ~ 1 0' ~ 1 0' ~ 1 0'~ 1 A study was then carried out on the problems associated with storage of bacteria.
The bacteria was studied in a dry state, a semi-dry state and a wet state to ascertain the change in their activity depending on such state and it was concluded that it was almost impossible to store the bacteria commercially for any length of time without adversely affecting the deodorizing ability thereof unless the bacteria are stored in the dry state. This conclusion was arrived at in view of the experimental data given below. Referring now to Table 14, the data in Table 14 was obtained using bacteria
FRI No. 2823 as a test sample. However, it has been found that similar data will be obtained if other similar bacterial strains are employed.
TABLE 14
Allowable period of storage State of bacteria Coating during storage Material 28 "C 8"C dry - 7 days 20 days semi dry - 5 days 15 days wet - 3 days 7 days Coexisting with liquor 2 Y3 days 5 days In Table 14, the dry state refers to a water content of 8%, the semi-dry state refers to a water content of 15%, the wet state refers to a wet-cake state in which the bacteria are merely filtered from the culture medium, and the "co-existing with liquor" state means that the bacteria are left in the culture medium at the end of cultivation. The allowable period of storage means the longest period of storage during which bacteria can be stored without decreasing their deodorizing power significantly.
It will be appreciated from the above that the water content of the bacteria and the temperature of storage were changed in fairly wide limits and it was found possible to prevent the deodorizing power decreasing after storage for an appreciable period of time. Since the inventor had found that S-, N- and C-compounds are very effective for prevention of the decrease of the deodorization power and also for achieving rapid growth in successive sub-cultures as described hereinbefore, it was decided to conduct experiments in which the bacteria having different water contents was coated with different coating materials. The results of these experiments are shown in Table 15(a) and 15(b) below.
TABLE 15-a
Allowable period -State of storage in storage Coating material 28 0C 80C S-, N- and C-compounds 25 days 120 days Mineral 10 days 40 days dry Vitamins 15 days 50 days Starch 20 days 100 days Cystine 20 days 100 days Glutamic acid 20 days 100 days S-, N- and C- compounds 5 days 25 days Mineral 4 days .15 days wet Vitamins 4 days 20 days Starch 4 days 25 days Cystine 4 days 25 days Glutamic acid 4 days 25 days S-; N- and C- compounds 4 days 20 days Mineral 3 days 12 days liquid Vitamins 3 days 12 days Starch 3 days 20 days Cystine 3 days 20 days Glutamic acid 3 days 20 days In Table 15 (a), -(b), the coating of the body of bacteria in the liquid state means that each coating material was added into the culture medium and intimately mixed, and then the mixture was allowed to stand for storage. Further, the length of the allowable period of storage differed considerably in accordance with the difference in the concentration of coating material and there existed an optimum concentration for the period of storage as shown in Table 15 (b). The data on the allowable period of storage shown in Table 15 (a) are those values obtained by the test on bacteria
F.R.I. No. 2823 coated with each coating material of the optimum concentration. The situation is the same also on those data which appear in Table 16 and Table 17.
TABLE 15 (b)
Allowable Period Concen- of Storage Strain State Coating tration (F.R.I. No.) in Storage Material (d) 280C 80C 1 2 days 10 days S- N- 5 3 days 15 days and C- compounds 10 3 days 20 days 20 4 days 20 days 2823 liquid 1 2 days 10 days 5 2 days 15 days Cystine 10' 3 days 20 days 20 3 days 20 days 1 2 days 10 days S-, N- and C-' 3 days 15 days compounds 10 4 days 20 days 20 3 days 12 days 2544 liquid 1 2 days 10 days 5 3 days 15 days Cystine 10 4. days 20 days 20 3 days 12 days 1 3 days 15 days S-, N- 5 4 days 20 days and C compounds 10 2 days 10 days 20 1 day 7 days 2545 liquid 1 3 days 15 days 5 4 days 20 days Cystine 10 3 days 12 days 20 2 days 7 days 1 4 days 20 days S-, N- 5 3 days 15 days and C- compounds 10 1 day 6 days 20 1 day 5 days 2546 liquid 1 4 days 20 days 5 4 days 15 days Cystine 10 2 days 10 days 20 1 day 5 days Remarks: O The concentration, d in the table, was defined as follows: l(d) means the liquid state contains 0.1% of each of Na2S .9H,O, ammonia and acetic acid in weight.
O The concentration of cystine, d in the table was also defined as
follows; l(d) means the liquid state contains 0.1% of cystine in weight.
Table 16 shows the results of storage of the said bacteria, wherein each bacteria
sample was coated with a material which exerts a noticeable effect in accelerating the
decrease of deodorizing power of the bacteria in successive sub-cultures.
TABLE 16
Allowable period of storage (days) State in Coating storage material 280C 80C dry Peptone 5 15 Broth 7 20 Tryptophan 6 20 Serine 7 20 wet Peptone 2 5 Broth 2 5 Tryptophan 2 5 Serine 2 7 liquid Peptone 2 5 Broth 2 4 Tryptophan 2 5 Serine 2 5 It will be appreciated from the above Tables, that in order to increase the allowable period of storage of the bacteria, the bacteria can be coated with a substance which is itself capable of retaining the original deodorization properties of the bacteria during sub-culture thereof. Thus, it can be deduced that there is a close relationship between substances which enable the original deodorization ability of the bacteria ta be retained in a sub-culture and a substance which preserves the deodorization properties of the bacteria during storage.
Faced with this relationship, further experiments were conducted to find bettel coating materials and the inventor has discovered that it is very effective to coat the bacteria with those materials which provide the most advantageous culture medium e.g. a combination of S-, N- and C-compounds and/or starch and/or vitamin + mineral or a combination of effective amino acids (See Tables 9 and 10). Table 17 (a) shows three examples of coating materials.
TABLE 17 (a)
Allowable period of storage State in storage Coating material 28 OC 8 0C A.. (S-W) - Glucose + F component 60 days 300 days + Starch + Mineral +Vitamins dry B .. Cystine + Cysteine + Methionine 40 days 200 days C .. S-, N- and C-' compounds + 60 days 300 days Starch + Mineral + Vitamins A 8 days 60 days wet B 7 days 50 days C 7 days 50 days A 6 days 60 days liquid B 5 days 50 days C 5 days 50 days Here, another example will be shown. The data shown in Tables 14, 15(a), 15(b), 16 and 17(a) for storage tests were obtained by sealing the sample in a nitrogen gas atmosphere; however, other gases can be effectively used. The effect of
N2, CO2, 2, H2, methane gas and air on the allowable period of storage are shown in Table 17 (b). From those data, it will be understood that CO2 and H2 are very superior to use for storage being almost comparable to nitrogen gas, however, air and methane gas are slightly inferior and oxygen gas is most inferior.
TABLE 17 (b)
Allowable pe'riod or storage State in Coating storage material Gas sealed 280C 8 C N2 60 days 300 days CO2 60 days 300 days A O2 15 days 50 days dry see Table 17(a) H2 55 days 250 days Methane gas 40 days 200 days Air 30 days 100 days N2 4Q days 200 days CO2 40 days 200 days B O3 12 days 40 days dry see Table 17(a) H2 40 days 200 days Methane gas 30 days 150 days Air 20 days 70 days N2 60 days 300 days CO2 60 days 300 days C O2 15 days 50 days dry See Table 17(a) H2 55 days 250 days Methane gas 40 days 180 days Air 30 days 100 days The following conclusions can be drawn from the above experiments.
(a) As far as the preservation temperature is concerned, a temperature of 80C Is
preferred to a temperature of 280C since the allowable period of storage of 80C
is about 5 times that obtained at 280C.
(b) It is not preferred to store the bacteria in the wet state or in the culture medium.
(c) Depending upon the bacterial strain concerned, there is an optimum coating
material concentration.
(d) If the bacteria are in a dry state and are coated with the correct coating material,
it is possible to store them for a long period of time whilst retaining the
deodorization properties of the bacteria.
(e) The most suitable coating material can be chosen depending upon the intended
use of the bacteria. For example, in the case where the bacteria are to be useS as a deodorizing agent by oral administration. a combination of an amino acid,
a vitamin, mineral and starch is suitable for the coating material whilst the use
of S-, N- and C-comDounds is to be avoided.
(f) There is a close relationship between the type of coating material employed and
the components of the culture medium which are effective to retain the deodoriz
ing properties of the bacteria, i.e. those components of the culture medium which
are effective to retain the deodorizing properties of the bacteria are most prefer
ably used, subject to paragraph (e) above for the coating material.
Furthermore, in order to produce a commercially viable deodorant product using the bacteria, it is necessary to use only a single strain a bacterium or a small number of strains. In the case where a large number of strains are employed, it is a matter of very great practical difficulty to ensure that the deodorizing power of many strains of bacteria are maintained constant and to produce a commercial product which is stable and of consistent quality. Moreover, in the case of the use of a very large number of strains, it is much more difficult to prove the non-pathogenicity of the resultant composition. Moreover, the use of a very large number of bacterial strains might upset the biological balance in nature. Thus, it is clear that the use of a single bacterial strain is most desirable and, if it is not possible to employ a single strain of bacteria, then it is necessary to use as small a number of bacterial strains as possible, e.g. two or three strains. However, the inventor has succeeded in isolating various strains which can be used alone or as a mixture of two or three for effective deodorization purposes. This is believed to represent a very important advance in the field of commercial use of bacteria for deodorization. Furthermore, the inventor has carried out development work in other areas which are important in the commercial use of bacteria. The first area is that the inventor has found that, when he has obtained bacteria of a single strain, such bacteria grow rapidly under aerobic or under facultative anaerobic conditions but are almost impossible to grow under anaerobic conditions. The importance of the ability of the bacteria to grow rapidly under anaerobic conditions has already been mentioned previously in the specification and, for a long time, the inventor tried to acclimatise the bacteria to anaerobic conditions since such conditions are encountered often in deodorization. The inventor has finally succeeded in obtaining bacteria of a single strain which can grow rapidly in anaerobic conditions and which have superior deodorization properties. The degree of multiplication of the bacteria just after isolation thereof into a single strain and that of bacteria after having been acclimatised to anaerobic conditions in anaerobic cultivation are shown in
Table 18.
TABLE 18
Count of bacteria (F.R.I.
No. 2823)/cc after an anaerobic culture at 280,C for 72 hrs.
Strain just Strain Culture medium after isolation acclimatized S-W + glucose ;+ F- component 1 x 108 3 x 109 S-W 5 x 108 5 x 109 S-W + Amino acid + Vitamins l 1 x 109 10 x The second area of research is the acclimatization of bacteria to a given temperature situated in a broad range of temperatures. This necessity, of course, comes from the fact that the deodorization bacteria will be used broadly everywhere in the wcrld, such as, from the tropical zone to subtropical, temperate zone and cold zone.
Thus, the bacteria of single strain should have a high rate of growth and also a strong deodorizing power at high temperatures and at low temperatures. If the temperature range is represented typically by three temperatures, such as + 80C, +28"C and +40"C, and the condition on the partial pressure of oxygen is classified into three, that is. aerobic, facultative anaerobic and anaerobic, 9 conditions appear as shown in the following table. To those conditions, the bacteria of single strain should be acclimatized.
+80C +280C +400C
aerobic 0 0 0
facultative anaerobic 0 0 0
anaerobic 0 0 0
Starting from a strain which did grow rapidly at 280C in the aerobic or facultative anaerobic conditions, a selective acclimatization to each of those 9 given conditions was carried out repeatedly for a long period and the inventor finally succeeded in cultivating them under these conditions.
The degrees of multiplication of those acclimatized bacteria and the degree of multiplication of the single original strain obtained by the isolation and used as the starting material for the acclimatization to the nine given conditi
TABLE 20
Residual odor Experimental condition Strain Strain (excrement) original acclimatized bacteria during spring 3 1' were added to a receiver during summer 3' 1' of excrement in a basin during autumn 3 1' during winter 4 1' 80C 2' 1 280C 1 0'- 1 400C 4 0' - 1 Remarks: O The original strain not acclimatized was that used in
Table 18.
O The acclimatized strains were chosen from those shown
in Table 19.
O The value of residual smell during each season is the
mean value during it.
Attention is drawn to the specification and claims of our co-pending British
Patent Application No. 21344/77 (Serial No. 1,584,694) which is concerned with a
Lactobacillus composition.
Claims (21)
WHAT WE CLAIM IS:-
1. A composition comprising only one or a small number of strains of autotrophic bacteria, said bacterial strain(s), when cultivated at 280C exhibiting a P (specific rate of growth) value of 0.25 more than that exhibited by colon bacteria in an S-W medium when the composition of the culture medium (Stephenson-Whetham) consists of 1 g of KH2POt, 0.7 g of MgSO, . 7H2O, 1 g of NaCI, 4 g of (NH4),HPO4, 0.03 g of FeSO4 . 7H,O and 5 g of glucose per litre of the culture medium; said bacterial strain, or said small number of strains in combination exhibiting accelerated growth in said culture medium in the presence of (a) each of Na3S . 9H2O, ammonia and a lower aliphatic carboxylic acid and/or (b) at least one of cystine, cysteine, and methionine.
2. A composition as claimed in claim 1, wherein said strain(s) exhibit(s) a strong resistance to gall.
3. A composition as claimed in claim 1 or 2, containing at least one bacterial strain exhibiting an S-action, at least one bacterial strain exhibiting an N-action, and at least one bacterial strain exhibiting a C-action, as major bacterial components.
4. A composition as claimed in claim 1 or 2, containing at least one set of two bacterial strains as follows:- (a) exhibiting S- and N-actions, and the other exhibiting C-action;
(b) one exhibiting N- and C-actions, and the other exhibiting S-action; or
(c) one exhibiting C- and S-actions, and the other exhibiting N-action, as major components.
5. A composition as claimed in claim 1 or 2, containing at least one bacterial' strain exhibiting each of S-, N- and C-actions, as a major bacterial component.
6. A composition as claimed in claim 1 or 2, wherein said bacterial strain(s) is or are FRI Nos.: 2544, 2545, 2546 and 2823, 3575, 3576, 3577 and 3578.
7. A composition as claimed in claim 1 or 2, wherein said bacterial strain(s) is or are a strain or strains subcultured in a subculturing medium containing (A) at least one of S-, N- and C-compounds and/cr (B) at least one S-containing amino acid.
8. A composition as claimed in claim 7, wherein the sub-culturing medium further comprises at least one of starch, minerals and vitamins.
9. A composition as claimed in claim 1 or 2, wherein said bacterial strain(s) is
or are strain(s) stored at a temperature of 80C.
10. A composition as claimed in claim 1 or 2, wherein said bacterial strain(s)
is or are strain(s) stored in CO2, N23 H2, He or an inert gas.
11. A composition as claimed in claim 1, wherein said bacterial strain(s) is or
are acclimatised to exhibit aerobic, facultative anaerobic or anaerobic properties.
12. A composition as claimed in claim 1, wherein said bacterial strain(s) is or
are acclimatised to grow at temperatures within the range of +8"C to 40"C, so as
to exhibit a high rate of growth within said temperature range.
13. A composition as claimed in claim 1, wherein the bacterial strain(s) exhíbit(s) a strong resistance to anti-bacterial agents and spices.
14. A composition as claimed in claim 1 or 2, wherein the bacterial cells are
coated with (A) a composition comprising (a) S-W medium minus glucose, (b)
one or more S-, N-, and C-compounds, (e) starch, (d) minerals and (e) vitamins,
(B) a composition comprising cystine, cysteine and merhionine or (C) a composition
comprising (a) one or more of S-, N- and C-compounds (b) starch, (c) vitamins
and (d) minerals.
15. A composition as claimed in claim 1 or 2, wherein the bacterial cells have
an ability to deodorise and are coated with a substance which contributes to the
retention of the deodorising ability of said bacterial cells when they are sub-cultured.
16. A composition as claimed in claim 1 or 2, wherein the bacterial cells are
coated with at least one material selected from the group consisting of S-, N- and
C-containing compounds, minerals, vitamins, starch, cystine and glutamic acid.
17. A composition as claimed in claim 1 or 2, wherein the bacterial cells are
coated with a superior culture medium for said cells; a combination of amino-acids
which are effective to preserve the deodorising power of said cells; at least one of a
combination of S-, N- and C-containing compounds, starch, and vitamins and
minerals; or a mixture thereof.
18. A composition as claimed in any one of claims 14 to 17, wherein the bacterial
cells are in the dry state.
19. A bacterial preparation comprising (I) a composition as claimed in any
preceding claim, and (II) a composition comprising (A) at least one of S-, N- and
C-compounds and/or (B) at least one S-containing amino acid.
20. A bacterial preparation as claimed in claim 19, wherein composition (II)
further comprises at least one of starch, a mineral and a vitamin.
21. A method for the deodorization of excrement which comprises contacting excrement with a composition or preparation as claimed in any preceding claim.
GB21343/77A
1976-05-21
1977-05-20
Bacterial deodorant composition
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Deodorizers and their manufacturing and storage methods
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1977-05-23
1977-05-23
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Bacterial deodorant composition
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1981-09-18
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Kao Corporation
Odor-removing and deodorizing composition employing a hydrolysate of keratin material
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DESODORING DEVICE.
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1991-02-26
Kurasawa Optical Industry Co., Ltd.
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Sederma Sa
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JP
JP51059474A
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patent/BR7703308A/en
unknown
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DE19772723226
patent/DE2723226A1/en
not_active
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1977-05-23
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FR7715857A
patent/FR2352058A1/en
active
Granted
1984
1984-04-30
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US06/604,417
patent/US4879238A/en
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1984-06-13
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(en)
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1981-09-18
1986-05-27
Kao Corporation
Odor-removing and deodorizing composition employing a hydrolysate of keratin material
EP0208818A1
(en)
*
1984-01-17
1987-01-21
Seikenkai Foundational Juridical Person
Biodeodorizer and the process for the preparation thereof
US4871539A
(en)
*
1984-01-17
1989-10-03
Seikenkai Foundational Juridical Person
Biodeodorizer and process for preparing same
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