PRODUCT FOR CLEANING, SANITIZING AND HYGIENIZATION

Abstract

Product for cleaning, sanification and hygienization, comprising a base solution with sanificant and/or hygienizing action to which probiotic bacteria and bacteriophage elements are mixed; the probiotic bacteria being preferably of the Bacillus subtilis, Bacillus megaterium and Bacillus pumilus genera, while the bacteriophage elements comprise, as an alternative or in combination, bacteriophages of the Caudavirales, Microviridae, Leviviridae, Inoviridae, Tectiviridae, Corticoviridae families.

Claims

1. Product for cleaning, sanification and hygienization, comprising: a base solution with detergent, sanificant and hygienizing action containing at least a surfactant chosen among nonionic surfactants, cationic surfactants and amphoteric surfactants; probiotic bacteria spores mixed to such base solution; further comprising bacteriophage elements mixed to such base solution and having bactericidal action on predetermined undesired bacterial species occurring on contaminated surfaces, such bacteriophages elements, in cooperation with the probiotic bacteria spores being able to carry out a combined and synergistic action against harmful microorganisms.

2. Product according to claim 1, wherein the concentration of nonionic surfactants is between 0.001% and 30%, the concentration of cationic surfactants is between 0.001% and 15%, and the concentration of amphoteric surfactants is between 0.001% and 15%.

3. Product according to claim 1, wherein the concentration of probiotic bacteria spores mixed to the base solution is between 10.sup.2 and 10.sup.9 spores/ml, preferably between 10.sup.6 and 10.sup.7 spores/ml.

4. Product according to claim 1, wherein the concentration of bacteriophage elements mixed to the base solution is between 10.sup.3 and 10.sup.9 PFU/ml, preferably between 10.sup.6 and 10.sup.7 PFU/ml.

5. Product according to claim 1, wherein the probiotic bacteria are of the Bacillus genus.

6. Product according to claim 5, wherein the probiotic bacteria are taken from the group consisting of the Bacillus subtilis, Bacillus megaterium and Bacillus pumilis species.

7. Product according to claim 1, wherein the bacteriophage elements comprise bacteriophages taken from the group consisting of the Caudovirales family, the Microviridae family, the Leviviridae family, the Inoviridae family, the Tectiviridae family, the Corticoviridae family and combinations thereof.

8. Use of a product for cleaning, sanification and hygienization according to claim 1 for treatment of surfaces.

9. Use according to claim 8 for treatment of boundary and/or furniture surfaces.

10. Product according to claim 2, wherein the concentration of nonionic surfactants is between 5 and 15%, the concentration of cationic surfactants is between 0.001% and 5%, and the concentration of amphoteric surfactants is between 0.001% and 5%.

Description

DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0032] The features of the invention will be highlighted in the following as a way of example in the description of some preferred, but not limiting embodiments.

[0033] The inventive, proposed product for cleaning, sanification and hygienization comprises: [0034] a base solution with sanificant and/or hygienizant action containing at least a surfactant chosen among the nonionic surfactants, cationic surfactants and amphoteric surfactants, [0035] probiotic bacteria spores mixed to such base solution; [0036] bacteriophage elements mixed to such base solution and having bactericidal action on predetermined undesired bacterial species, such bacteriophage elements in cooperation with probiotic bacteria spores being able to carry out a combined and synergistic action against harmful microorganisms.

[0037] Nonionic surfactants can occur in the base solution with a concentration between 0.001-30%, preferably between 5-15%.

[0038] Cationic surfactants can occur in the base solution with a concentration between 0.001-15%, preferably between 0.001-5%;

[0039] Amphoteric surfactants can occur in the base solution with a concentration between 0.001-15%, preferably between 0.001-5%.

[0040] The base solution can contain one or more of said nonionic, cationic and amphoteric surfactants in the indicated concentrations.

[0041] Probiotic bacteria spores can occur in the base solution with a concentration between 10.sup.2-10.sup.9 spores/ml, preferably between 10.sup.6-10.sup.7 spores/ml.

[0042] Bacteriophage elements specifically directed against bacteria persistent on the contaminated surfaces can occur in the base solution with a concentration between 10.sup.3-10.sup.9 PFU/ml, preferably between 10.sup.6-10.sup.7 PFU/ml.

[0043] Concerning the probiotic bacteria, as a way of not limiting example, they can be of the Bacillus genus, for example they can be the Bacillus subtilis, Bacillus megaterium and Bacillus pumilus species.

[0044] It is clear that any other probiotic bacteria can be used according to the undesired bacterial species to be fought.

[0045] Advantageously the bacteriophage elements can be chosen, as an alternative or in combination, according to the bacterial species to be fought among the following families: Caudovirales, Microviridae, Leviviridae, Inoviridae, Tectiviridae, Corticoviridae.

[0046] As it is known the bacteriophage elements of Caudovirales family comprise also the Myoviridae, Siphoviridae, Podoviridae families.

[0047] The proposed product for cleaning, sanification and hygienization, in the preferred above described embodiments is particularly indicated for treatment of surfaces and/or ambients, comprising the treatment of inner surfaces of pipes for circulation of fluids for food use (for example water).

[0048] The action of bacteriophages is specific, since specific types of lytic bacteriophages kill specific types of bacteria, thus carrying out a potentially very efficient and targeted action in fighting directly against undesired bacteria.

[0049] It is also to be highlighted how the use of specific bacteriophages avoids the risk of damage of Bacillus probiotics, which thus can keep their activity intact and efficient.

[0050] The use of bacteriophages in products for cleaning, sanification and hygienization is particularly indicated since they are quite stable in different conditions of temperatures, pH and environmental salinity, and so they can be added to detergents diluted without loosing activity.

[0051] Lab experimental tests have been carried out which highlight the synergistic effect deriving from the combined use of probiotic bacteria and bacteriophage elements in comparison to the single use of probiotic bacteria or bacteriophage elements.

[0052] Test n? 1 (Single Use of Probiotic Bacteria)

[0053] The activity of PCHS detergents containing probiotics has been widely evaluated in the field, where it has shown to be able to reduce the microbial count of about 90% more with respect to the traditional detergents, by means of daily use extended for about 1 month.

[0054] After a period of 2 months the reduction of contaminant count is stably low both for what concerns bacteria and fungi [Caselli et al., 2016].

[0055] In the following figures, it is to be noted the antimicrobial action (bacteria and fungi) of the detergent based only on probiotics.

[0056] The development of contamination by Staphylococcus aureus (as example of gram-positive bacteria), Enterobacteriaceae spp. (as example of gram-negative bacteria) and Candida albicans (as example of fungi) was measured in the field, by application of Rodac plates of culture medium specific for the bacterial and fungi species indicated.

[0057] Test n? 2 (Single Use of Bacteriophages)

[0058] In the following figures, it is to be noted the antibacterial action of bacteriophages on surfaces contaminated by specific target bacteria.

[0059] The development of contamination by Staphylococcus aureus (as example of gram-positive bacteria), Pseudomonas aeruginosa (as example of gram-negative bacteria) and Candida albicans (as example of fungi) was measured in vitro, on experimental models of surfaces of nonporous and sterile material (single fired ceramics 24 cm.sup.2).

[0060] The surfaces have been contaminated with a known quantity of microorganism (10.sup.2 CFU/24 cm.sup.2) and in the following treated by application of increasing concentrations of specific bacteriophages (10.sup.3, 10.sup.4, 10.sup.5 PFU respectively) with ratio 10:1, 100:1 and 1000:1 to the target bacteria in culture.

[0061] For fungi it was used the combination of bacteriophages used against gram positive and gram negative bacteria.

[0062] The residual contamination was evaluated after 1, 3, 6 hours by application of Rodac plates of culture medium specific for the bacterial and fungi species indicated.

[0063] The results are expressed as average values?S.D. of samples in duplicate in three independent experiments.

[0064] It can be noted that already after 1 hour, and at the lowest concentration, the bacteriophages are able to eliminate more than 90% of bacterial cells against which they are specifically directed.

[0065] The treatments with only bacteriophages has the drawback to be directed against only bacteria specifically recognized by bacteriophages, and so in case of contamination by bacterial species other than the ones against which the bacteriophages are specifically directed, a re-contamination of the treated surface can occur which is due to the fact that the bacteriophages are, by definition, specific for a bacterial species, and so not able to attack other bacterial species, much less to attack fungi species.

[0066] These drawbacks of bacteriophages are overcome by adding probiotics, which have instead a slower but generalized action regardless of the bacterial or fungi species occurring.

[0067] Test n? 3 (Combined Use of Probiotic Bacteria and Bacteriophages)

[0068] In the following figures, it is to be noted the effect of the combined and synergistic antimicrobial action obtained by the contemporaneous occurrence of probiotic bacteria and bacteriophages on surfaces contaminated by the specific microbes shown.

[0069] The development of contamination by Staphylococcus aureus (as example of gram-positive bacteria), Pseudomonas aeruginosa (as example of gram-negative bacteria) and Candida albicans (as example of fungi) was measured in vitro, on experimental models of surfaces of nonporous and sterile material (single fired ceramics 24 cm.sup.2).

[0070] The surfaces have been contaminated with a known quantity of microorganism (10.sup.3 CFU/24 cm.sup.2) and in the following treated by application of a solution containing probiotic bacteria (10.sup.3 PFU/ml) and specific bacteriophages (10.sup.4 PFU/ml).

[0071] For fungi it was used the combination of bacteriophages used against gram positive and gram negative bacteria.

[0072] The residual contamination was evaluated after 1, 2, 3, and 4 weeks by application of Rodac plates of culture medium specific for the bacterial and fungi species indicated. The results are expressed as average values?S.D. of samples in duplicate in three independent experiments.

[0073] Comparison of the Tests Carried Out

[0074] In the following figure, it is shown the direct comparison of the antimicrobial action of the three treatments on the contamination by Staphylococcus aureus, measured in vitro on experimental models of surfaces of nonporous and sterile material (single fired ceramics 24 cm.sup.2).

[0075] The surfaces have been contaminated with a known quantity of S. aureus (10.sup.3 CFU/24 cm.sup.2) and in the following treated by application of a solution containing only probiotic bacteria (10.sup.3 PFU/ml), only specific bacteriophages (10.sup.4 PFU/ml), or the combination of probiotics and bacteriophages at the concentrations indicated.

[0076] The residual contamination was evaluated after 1, 2, 3, and 7 days by application of Rodac plates of culture medium specific for Staphylococcus aureus.

[0077] The results are expressed as average values?S.D. of samples in duplicate in three independent experiments.

[0078] From the experimental tests carried out, it is to be noted easily how the contemporaneous occurrence of Bacillus probiotics and anti-pathogenic bacteriophages increases remarkably the efficacy of the product for cleaning, sanification and hygienization, thanks to their synergistic action carried out against the various harmful microorganisms.

[0079] On the one hand bacteriophages reduce the contaminant count extremely rapidly but they have a time limited action and are not able to avoid re-contamination.

[0080] On the other hand, probiotics reduce the contaminant count slowly by constantly, thanks to the competitive mechanisms, thus inhibiting the re-contamination.

[0081] The contemporaneous occurrence of probiotics and bacteriophages guarantees the rapidity of antimicrobial action and at the same time the inhibition of re-contamination, thus guaranteeing a rapid and time stable reduction of pathogens.

[0082] The realization of a product for cleaning, sanification and hygienization which contains at the same time Bacillus probiotics and anti-pathogenic bacteriophages is clearly advantageous with respect to the traditional products, in that it guarantees an extremely rapid and immediate reduction of the contaminant bacterial count.

[0083] This is due to the combined and synergistic action carried out by the bacteriophage elements which are able to kill the target bacteria in few hours, and by probiotic bacteria which are able to colonize the objects treated persistently, by taking rapidly the place of pathogens.

[0084] Therefore the occurrence of bacteriophages represents a strengthening of the antibacterial action of the traditional product based on Bacillus probiotics with respect to the pathogens, thus easing and increasing the action of probiotics, which result active on fungi as well.

[0085] This guarantees an immediate effect in the initial step of cleaning, sanfication and hygienization treatment, thus allowing the probiotic bacteria to determine, in faster times than the traditional products, a low, stable and lasting in time presence of potentially pathogenic bacteria in the ambients and objects treated.

[0086] The addition of specific bacteriophage elements to the base solution mixed to probiotic Bacillus spores allows advantageously to use the product in targeted way, answering to specific needs and/or situations of particular interest, as for example the reduction of one or more particularly occurring and/or harmful specific bacterial species.

[0087] The occurrence of specific bacteriophage elements represents another safety means concerning the use of Bacillus probiotics bacteria since above all in the initial steps of attack, those microorganisms are destroyed which are potentially able to give to Bacillus undesirable characters by means of gene exchange.

[0088] The presence in the inventive product of a greater number of bacteriophage elements families (Caudovirales, Microviridae, Leviviridae, Inoviridae, Tectiviridae and Corticoviridae) allows the same product to have a bactericidal action on a greater group of treatable bacterial species.

[0089] The predetermined families of bacteriophage elements can be present in the product object of the invention both singularly and in combination, so that it is contemplated the possibility of diversified treatments according to the specific need.

[0090] From what above described it is clear how the proposed product for cleaning, sanification and hygienization is able to act against any microbial species, of the multi-resistant type as well (superbugs), in a particularly rapid and efficient way with respect to the traditional products indicated in the preamble.

[0091] The proposed product for cleaning, sanification and hyginization by comprising a base solution with sanificant and/or hygienizing action to which probiotic bacteria and bacteriophage elements are mixed, has decidedly an extremely reduced environmental impact.

[0092] Such product is particularly apt for treatment of surfaces and/or ambients comprising the treatment of inner surfaces of pipes for circulation of fluids for food use (for example water).