SANITIZING DISINFECTANT COMPOSITION

Description

SANITIZING DISINFECTANT COMPOSITION 1. State of the art In recent years, dental caries and biofilm formation have become among the most common infections, and improper management leads to the development of serious diseases and the formation of biofilm. Oral hygiene is an important measure for the prevention of oral diseases (dental caries and periodontitis) and a prophylactic treatment for aspiration pneumonia, type 2 diabetes mellitus, and cardiovascular disease. Although dental caries is a global health problem, it is preventable with appropriate interventions, especially in the early stages of life. However, some groups cannot easily perform regular oral hygiene care (brushing or flossing) on ​​their own, including seriously ill patients, the elderly, and those displaced by disasters. Furthermore, many individuals lack the knowledge and motivation to maintain oral hygiene.Failure to maintain oral hygiene leads to the formation of microbial biofilms, called dental plaque, on tooth surfaces. The formation of biofilms on tooth surfaces by cariogenic bacterial communities is the initial step in the development of dental caries. Streptococcus mutans is a primary etiological agent of dental caries. The main virulence traits associated with S. mutans cariogenicity are acid production from fermentable dietary carbohydrates, acid tolerance, and exopolysaccharide (EPS) formation. Acid production promotes the demineralization of tooth enamel, and acid tolerance allows survival in the low-pH environment within dental plaque. EPS promotes the formation of acid-producing biofilms on tooth surfaces, which are bioaggregates resistant to mechanical tooth brushing.Although fluoride preparations protect tooth surfaces from acid attacks, their effects are limited if not combined with plaque control. The use of bactericidal compounds to eradicate cariogenic bacteria is controversial because these compounds disrupt healthy oral microflora and can lead to the development of multidrug-resistant bacteria. Oral formulations currently in use are well-known and come in many different forms. They are often used for dental care or to prevent dental disease and come in any physical form, such as liquid, solid, gaseous, aerosol, spray, nebulizer, and combinations thereof to create pastes, gels, toothpastes, mouthwashes, delivery devices, and more. For example, well-known oral formulations are typically a mixture of alcohol and / or water and often contain other ingredients. Alcohol, such as ethanol or propanediol, can be used for disinfection and / or preservation. Furthermore, many substances frequently used in oral formulations dissolve much better in an alcohol-water mixture than in pure water, so alcohol can also serve as a solubilizer. For example, antiseptic active ingredients such as chlorhexidine are added to well-known oral formulations, some of which dissolve much better in alcohol-water mixtures than in pure water. Furthermore, some well-known oral formulations contain fluorides, which are intended to harden enamel. In terms of composition and mode of action, these well-known oral formulations are generally primarily oriented toward a curative approach. Although a disinfectant effect, i.e., an overall reduction in the germ or bacterial load, is also expected and desired, it plays only a secondary role. However, numerous oral diseases, especially inflammatory ones, ranging from simple gingivitis to periodontitis and peri-implantitis, to childhood and adult caries, also require a disinfectant approach. Indeed, peri-implantitis results from the action of a pathogenic noxa in the abnormal presence of certain bacterial species and their metabolic products (biofilms), in which a germicidal or disinfectant effect is desirable. Antibiotics are considered one of the most important weapons in the fight against these bacterial infections. Since their introduction, they have significantly improved the quality of human life in terms of health. However, these health benefits have been threatened in recent decades, as many commonly used antibiotics have become less effective against certain diseases, not only because many of them cause toxic reactions, but also due to the emergence of drug-resistant bacteria. The development of resistance is a serious problem today, as bacterial resistance is often not limited to the specific antibiotic prescribed to the patient but can generally extend to other compounds in the same class. Bacterial resistance and its rapid increase are a major concern for global public health and are emerging as one of the major challenges to human health. Treating bacterial infections with antibiotics is therefore useful, but their widespread use has led to alarming resistance among microorganisms and, ultimately, the recurrence of old infectious diseases. Therefore, an antibiotic-based approach to decimating bacterial burdens in the human oral cavity is undesirable, especially for chronic or persistent disease models. Added to this is an unexpected event involving the molecule chlorhexidine, considered the golden standard for oral antibacterials. In fact, a study published in Antimicrobial Agents and Chemotherapy highlights that exposure to increasing concentrations of chlorhexidine, a component widely used in disinfectants and antiseptics for domestic and hospital use, leads to the emergence of strains of Klebsiella pneumoniae that are also resistant to colistin, considered the latest frontier in antibiotic treatment for multidrug-resistant forms. Chlorhexidine, discovered by chance in Manchester in the ICI laboratories where new antimalarials were being researched, has had a successful life. After a modest start in operating rooms as a disinfectant (Hibitane and Hibiscrub), by the late 1970s, publications on its use in dentistry numbered just a few dozen. After that, the boom: mouthwash, gel, toothpaste; it seemed like a never-ending success, denying any space to aspiring competitors (who still remembers Sanguinaria canadensis?). The eternal battle against oral pathogens seemed to be coming to an end. In fact, Harald Loe himself, one of the pioneers of oral use of chlorhexidine, was the first to warn of the risk of resistance as early as 1972. Now the risk is a certainty. Unlike antibiotics, where the level of concern is almost hysterical given the alarming level reached, the problem of antiseptic resistance is much less well-known (and feared), especially with regard to chlorhexidine. For others, such as triclosan, authorities in both the US and Europe have already intervened, banning its use in personal hygiene products (see box on this page). For a couple of years, however, chlorhexidine has been under special surveillance, suspected of inducing cross-resistance against colistin, a valuable antibiotic used against the most resistant bacteria. In the near future, therefore, it is likely that the dosage and indications for chlorhexidine will be restricted, similarly to antibiotics. Perhaps we will see it disappear from toothpastes and soaps, while gels and mouthwashes will be used more selectively and for limited periods in patients where there is a demonstrated benefit.The same problem of bacterial resistance also arises for other oral antiseptics such as cetylpyridinium chloride, towards which the appearance of resistant strains in proportion to exposure has been demonstrated in vitro. Shortly after the FDA banned triclosan and 17 other compounds from personal hygiene products, an interesting article was published in the journal of the American Society for Microbiology in which the authors exposed the serious environmental changes caused by the unjustified use of antiseptics. According to the reported data, triclosan is the second most common chemical compound found in urban wastewater treatment plants, ahead of pharmaceuticals and surfactants. First place? Triclocarban, another antiseptic banned by the FDA. Considering that solid wastewater residues are used in agriculture as fertilizer, a few extra concerns certainly aren't alarmist and should make everyone think twice before purchasing a household product advertised as a "sanitizer." The first known use of chlorhexidine in dentistry dates back to 1959, when it was employed as an endodontic disinfectant. Its ability to inhibit tooth decay in animals was subsequently discovered, and in 1970, Harald Loe's famous study on dental students who had been deprived of their toothbrushes was published. In the group that, fortunately, used 0.2% chlorhexidine mouthwash as their only form of oral hygiene, their teeth remained plaque-free, unlike the control group. In 1972, the same authors published the first reports of strains of S. sanguinis showing reduced sensitivity after prolonged use of chlorhexidine mouthwash, but this finding was not considered clinically significant for many years. In 1980, it was discovered that resistance was a genetically transmitted trait.A few years ago, some authors found strains in dental plaque with reduced sensitivity to chlorhexidine and resistant to many antibiotics, warning against the prolonged use of oral hygiene products containing this disinfectant. 2. Summary of the invention The invention is therefore based on the task of specifying an oral composition that, for its intended use in the human mouth and oral cavity, has a germicidal effect with a high degree of tolerance for humans or human tissues. Furthermore, it should have zero environmental impact. As regards the oral composition, this task is solved according to the invention since it is formed by a composition which presents as ingredients, in addition to various excipients aimed at achieving the desired variable composition, a sugar (D-Tagatose) and a trace element (Copper) which is well-established in use as a food additive recognised and authorised worldwide thanks to a unique safety profile and an effective anti-biofilm activity. Furthermore, to date, there are no reports of the emergence of strains resistant to D-Tagatose or Copper, unlike what occurs with chlorhexidine. The invention is based on the idea that the desired germicidal effect of the oral composition can be achieved by specifically providing an alternative agent for the treatment of infectious diseases, avoiding the use of antibiotics and the associated risk of resistance development. This latter approach, namely combination or synergistic therapy against resistant microorganisms, may lead to new ways of treating infectious diseases and will likely be a potential area for further investigation in the future. The mode of action of combination therapy differs significantly from that of the drugs themselves acting alone; therefore, selecting an appropriate combination is crucial and important and requires an understanding of the potential interactions and synergistic effects. The advantages of this approach are: greater efficacy, reduced side effects, greater stability or bioavailability of the free active ingredients, and the achievement of an adequate therapeutic effect at relatively low doses compared to a synthetic drug. Considering that the oral mucosa is naturally absorbed (see sublingual anti-inflammatory tablets, trinitrine, and others), it is undesirable to introduce potentially toxic substances into the oral cavity as has been done to date. Instead, the use of food additives, which are virtually harmless to humans, including D-Tagatose and Copper, is preferred. Surprisingly, it was discovered that D-Tagatose and Copper, preferably in their colloidal form, especially when appropriately combined, not only enhance the effect of the individual elements, but also ensure significant product stability after packaging. In a particularly advantageous and independently inventive project, the use of the oral composition described above is envisaged for reducing the germ load in the human mouth or pharynx and / or for the care of teeth or devices used in the oral cavity and surrounding anatomical tissues. The advantages achieved with the invention lie in the fact that the proposed combination of D-Tagatose and a trace element, specifically copper, allows for particularly well-tolerated and equally effective reductions in germ or bacterial counts in the oral cavity, while avoiding the use of antibiotics and the development of resistance that can be triggered. Surprisingly, the composition exhibits synergistic effects, allowing for particularly effective reduction of bacterial counts and the associated treatment of inflammation resulting from infections. 3. Summary For its intended use in the human mouth and pharynx, an oral composition must have a significantly greater germicidal effect than known oral compositions, with a high degree of tolerance for humans or human tissues and with the exclusion of the possible formation of resistance. To this end, according to the invention, an oral composition is provided, comprising as its components a sugar and a trace element. Preferably, a combination of copper and D-Tagatose is provided as an antimicrobial agent active against biofilm. 4. Summary of the invention The development of specific behavioral measures for cariogenic bacteria should reduce dental plaque formation. Several studies have been conducted to determine whether the growth and adhesion of S. mutans are inhibited by various natural products, such as green tea catechins, cranberry constituents, citrus lemon oil, and mushroom extracts. However, these studies have not assessed biofilm formation. Sugar alcohols (polyols) are alternative candidates used for the prevention of dental caries. Rare sugars are generally monosaccharides, and their derivatives are rarely found in nature. Recently, rare sugars have received attention as healthy sugar substitutes due to their equivalent sweetness but much lower calorie content than sucrose. These sugars are thought to reduce calorie intake, thereby decreasing the risk of type 2 diabetes mellitus and obesity.The ketohexose D-tagatose has 92% of the sweetness but 38% of the calories of sucrose. D-tagatose is not a preferential substrate for bacterial fermentation, and it has been reported that D-tagatose is not readily catabolized by many lactic acid bacteria or pathogens such as Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, Bacillus cereus, and Yersinia enterocolitica. D-tagatose has been shown to suppress the growth of aerobic lactic acid bacteria in minced ham, thereby extending the shelf life of these products by 7–10 days. These results indicate that foods containing D-tagatose can suppress oral cariogenic bacteria. Indeed, it has recently been reported that D-tagatose inhibits acid production, growth and water-insoluble glucan production of S. mutans GS-5 in the presence of sucrose.The inhibitory effects of D-tagatose on the growth and biofilm formation of S. mutans GS-5 were examined. Monitoring S. mutans growth over a 24-hour period revealed that D-tagatose prolonged the lag phase without interfering with final cell yield. This growth delay was observed even in the presence of 1% sucrose, although it was abolished by the addition of D-fructose. S. mutans biofilm formation was significantly inhibited by growth in sucrose media supplemented with 1% and 4% D-tagatose compared to that in a culture containing sucrose alone, whereas S. mutans formed granular biofilms in the presence of this rare sugar. The inhibitory effect of D-tagatose on S. mutans biofilm formation was significantly more pronounced than that of xylitol. Growth on sucrose-based media supplemented with D-tagatose significantly reduced the expression of glucosyltransferase, exo- and fructosidase, and D-fructose-specific phosphotransferase genes, but not fructosyltransferase expression compared to the culture containing sucrose alone. Cell-associated glucosyltransferase activity in S. mutans was inhibited by 4% D-tagatose. These results indicate that D-tagatose reduces the production of water-insoluble glucan from sucrose by inhibiting glucosyltransferase activity. Colloidal copper is commonly assumed to have antibacterial properties.Copper generates toxic hydroxyl radicals that damage the cell membranes of Gram-negative and Gram-positive bacteria, including Enterococcus faecalis, present in infected root canals. Furthermore, the use of certain polymers to stabilize colloidal copper increases its release time and can also reduce the risk of bacterial recolonization and biofilm formation within the canals, enhancing the antimicrobial properties of these compounds. Copper is an essential element for the metabolism of animal and plant cells. It is a trace element present in most organisms, and more than 30 types of proteins contain it. Copper was the first metal used by humans over 9,000 years ago. Today, global copper consumption is approximately 18 million tons per year. As early as 2200 BC, it was used to sterilize wounds and drinking water.In the 19th century, copper workers were observed to be immune to cholera, and its use became widespread in the 20th century. Laboratory and clinical studies have been conducted to demonstrate its effectiveness. In recent years, copper has been used as a construction material for hospitals and medical centers, as its antimicrobial properties have proven useful in fighting infections. Colloidal copper, due to its antimicrobial properties, is important and has potentially promising applications in the fight against the growing number of pathogens resistant to currently available antimicrobials. These pathogens pose a continuing threat to human and animal health (Usman et al., 2013 & Kruk et al., 2015). The goal of our studies was to evaluate and assess the efficacy of colloidal copper and D-tagatose in combination, creating a synergistic effect with the resulting reduction of biofilm bacterial load and improving dentin sensitivity due to gingival recession. This solution could be best expressed through the use of a gel, but also in other forms such as chewing gum, sweets, and mouthwashes, allowing for direct application to the anatomical sites involved in the etiopathogenesis of biocide formation on the tooth surface. Applications could be varied, from caries prevention in pediatric patients and beyond.Other, but not exclusive, applications include orthodontic and prosthetic dental appliances, and the prevention of biofilm formation on implant abutments, a frequent cause of implant-based prosthetic therapy failure. The new composition not only has an immediate synergistic effect on bacterial biofilm formation, but also an unexpectedly long-lasting effect, making it a fundamental and effective aid, maintaining its effectiveness even during nighttime hours against the cariogenic action of biofilm. To date, there are remedies with a limited effect over time, often of chemical-synthetic origin, but which have never demonstrated a lasting and effective action. Furthermore, the presence of copper guarantees a marked action against antibiotic-resistant bacteria and to date there are no publications demonstrating the development of a copper-resistant action by bacteria belonging to the Streptococcus mutans category. Furthermore, for the first time, a composition has been identified with active ingredients classified as food additives and therefore endowed with a safety and tolerability profile never achieved with products of synthetic chemical origin and their degradation products, both identifiable as known impurities and undetectable as unknown impurities. In order to corroborate the theoretical assumption, an in vitro validation study was undertaken. 5. Experimentation 5.1. Experiment on the synergistic efficacy of colloidal copper and D-tagatose Experiment No. 1909 / 2023: Streptococcus mutans strain ATCC 25175 / Experimental preparations of D-Tagatose, colloidal copper > 100 nm. Combination of D-Tagatose and colloidal copper > 100 nm. Date: 14.09.2023-16.09.2023 Objective: Determination of the antimicrobial activity of preparations of D-Tagatose, colloidal copper > 100 nm. and a combination of D-Tagatose and colloidal copper > 100 nm. Materials: concentrated preparations of aqueous solutions: N. 6-8-1 - concentrated preparation of D-Tagatose, colloidal copper > 100nm, N. 6-8-2 - concentrated preparation of D-Tagatose; N. 6-8-3 - concentrated preparation of colloidal copper > 100nm; culture broth produced; mannitol - agar salt ; Test strain: 1. Streptococcus mutans strain ATCC 25175 Methods: A suspension of the test strains with a density of 0.5 McFarland was prepared using a Densi-La-Meter instrument. Further dilution of the preparations was done with nutrient broth 1:1. Antimicrobial activity was determined by incubation (exposure) at 37°C for 30 and 90 minutes for the original preparations and for 3 hours and 24 / 48 hours for the 1:1 diluted preparations using the macro method, followed by inoculation onto a dense culture medium. 106 microbial cells / ml of the suspensions of the test strains were added to the prepared solutions, as well as to the control tube. Controls consisted of distilled water without antimicrobial substances with the culture of the test strain (for original preparations) and nutrient broth diluted 1:1 with distilled water without antimicrobial substances with the culture of the test strain (for diluted preparations). Results: Test strain control: Streptococcus mutans, ATCC 25175: inoculation in 30 minutes - 106 microbial cells / ml; inoculation in 90 minutes - 5 * 106 microbial cells / ml; inoculation in 3 hours - confluent growth of the test strain; within 24 hours, a marked increase in broth turbidity is observed. Table 1 - Determination of the antimicrobial activity of aqueous solution preparations against the test strain Streptococcus mutans strain ATCC 25175 Colloidal copper > 100nm. Preparation Concentration Growth results of the test strain after incubation with the preparations Inoculation on dense culture medium after incubation Visual recording in a liquid nutrient medium 30 min 90 min 3 hours incubation 24 / 48 hours 6-8-3 Colloidal copper > 100nm. 99.99 % inhibition 10 CFU 99.9999 % inhibition 10 CFU 99.9999 % inhibition clear / turbid Table 2 - Determination of the antimicrobial activity of aqueous solution preparations against the test strain Streptococcus mutans strain ATCC 25175 D-Tagatose Preparation Concentration Growth results of the test strain after incubation with the preparations Inoculation on dense culture medium after incubation Visual recording in a liquid nutrient medium 30 min 90 min 3 hours incubation 24 / 48 hours 6-8-2 D-Tagatose 99.99 % inhibition 10 CFU 99.9999 % inhibition 10 CFU 99.9999 % inhibition turbid / turbid Table 3 - Determination of the antimicrobial activity of aqueous solution preparations against the test strain Streptococcus mutans strain ATCC 25175 Colloidal copper + DTagatose Preparation Concentration Growth results of the test strain after incubation with the preparations Inoculation on dense culture medium after incubation Visual recording in a liquid nutrient medium 30 min 90 min 3 hours incubation 24 / 48 hours 6-8-3 Colloidal copper + D-Tagatose no growth 100.00% inhibition no growth 100.00% inhibition no growth 100.00% inhibition clear / clear 5.2. Conclusions In this study, all preparations demonstrated an antimicrobial effect, but to varying degrees. They can be classified based on their efficacy: No. 6-8-1 > No. 6-8-2 > No. 6-8-3. 6. Detailed description 6.1. Colloidal copper as an antibacterial. Copper has a bactericidal effect, primarily due to its ability to donate and accept electrons in a continuous process. It produces a hydroxyl radical that can participate in a series of adverse reactions to cellular macromolecules, such as the oxidation of proteins and lipids. The hydrogen peroxide generated can lead to increased production of toxic hydroxyl radicals. It causes the displacement of iron from iron-sulfur clusters and can compete with zinc or other metal ions, important for protein binding sites; it disrupts membranes, denatures DNA, and blocks cellular respiration. In turn, bacteria have developed copper-tolerant mechanisms, such as extracellular sequestration of the ion, impermeability of external and internal bacterial membranes, removal of copper proteins (metallothionein) in the cytoplasm and periplasm, and copper efflux from the cell (Grass et al., 2011).Copper alloys acting on abiotic surfaces have been shown to cause, in addition to the rapid killing of antibiotic-resistant strains, the destruction of plasmid and genomic DNA, which has implications for preventing the spread of infections and gene transfer (Warnes et al., 2013 & Bagchi et al., 2013). Copper surfaces or their alloys are capable of eliminating 99.9% of pathogenic bacteria within hours, including methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, Pseudomonas aeruginosa, Listeria, and bacterial DNA lesions (Santo et al., 2011). The starting point for the clinical use of copper, specifically for disinfection of the root canal system, is reported in an ex vivo study in which canals treated with copper sulfate pentahydrate showed a 6-logarithm reduction in colony-forming unit counts on day 4 (Sànchez-Sanhueza et al., 2015). 6.2. Tagatose as an antibacterial Effects of D-tagatose on the growth of S. mutans GS-5. The effects of D-tagatose on the growth of S. mutans GS-5 were examined in BHI containing 1% sucrose. Sucrose increased the growth of S. mutans GS-5 compared to BHI alone, and the pH of the sucrose-containing culture dropped to less than 5.0 after 9 hours of incubation. Interestingly, D-tagatose delayed the transition of S. mutans growth to logarithmic phase despite the presence of sucrose. Correspondingly, the decline in culture pH was also delayed by D-tagatose compared to sucrose alone. This growth delay became more evident and entry into stationary phase was delayed by 6 hours when the D-tagatose concentration was increased to 4%; However, no significant differences in final OD590 were observed after 24 h of cultivation (data not shown). As demonstrated by the growth retardation of S.mutans GS-5 and the pH decline induced by D-tagatose were abolished when 1% D-fructose, but not D-glucose, was added to BHI containing 1% sucrose. These results indicate that the rare sugar D-tagatose inhibits sucrose catabolism in S. mutans GS-5. Effects of D-tagatose on S. mutans GS-5 biofilm formation in vitro. Because sucrose metabolism is important for the production of water-insoluble glucan, which is required for biofilm formation, D-tagatose was predicted to inhibit S. mutans biofilm formation. The effects of D-glucose, xylitol, and D-tagatose on S. mutans GS-5 biofilm formation in vitro were therefore evaluated. Addition of 1% sucrose to growth media significantly increased biofilm formation by S. mutans GS-5 compared to unsupplemented media (P<0.05), which is consistent with many previous reports.Supplementation with D-glucose and sucrose slightly reduced biofilm formation of S. mutans GS-5 compared to unsupplemented media. S. mutans biofilm formation was significantly reduced compared to sucrose alone (P2%). In contrast, D-tagatose showed a clear dose-dependent inhibition of S. mutans GS-5 biofilm formation. To determine whether the effects on biofilm formation were caused by high osmolality, biofilm formation by S. mutans GS-5 was compared in the presence of 1% and 5% sucrose. The biofilm mass in the culture with 5% sucrose was significantly smaller than that in 1% sucrose (OD550 1.27±0.05 vs. OD550 1.09±0.07, respectively; P<0.01 (data not shown). However, the addition of 4% D-tagatose to the 1% sucrose culture reduced the biofilm mass to nearly half that of the 5% sucrose culture (OD550 0.57±0.06 vs. OD550 1.09±0.07, P<0.01; data not shown). These results indicate that the effects of high osmolality were limited under the conditions used in this study. Scanning electron microscopic examination of S. mutans GS-5 biofilms. S. mutans GS-5 was grown in 1 ml of BHI containing 1% sucrose with or without 1 or 4% xylitol or D-tagatose in 24-well plates with plastic disc inserts; The plates were incubated anaerobically at 37°C for 72 hours, and the biofilms formed on the plastic discs were compared. S. mutans GS-5 grew equally well on all tested media. However, less biofilm formed on the discs in the cultures.containing D-tagatose compared to those in other media (1% sucrose alone or 1% sucrose plus 1 or 4% xylitol). In particular, multiple cellular aggregates of S. mutans GS-5 were observed in the cultures containing D-tagatose, especially at the highest concentration, while in the other cultures containing 1% sucrose, homogeneous biofilms formed on the discs. SEM examination of the discs also revealed a lower presence of biofilms on the discs in the culture with D-tagatose. Quantification of S. mutans GS-5 biofilms on the discs showed a significant reduction in the presence of D-tagatose (P<0.05). The use of non-cariogenic sweeteners is a method of preventing dental caries, and sugar alcohols such as xylitol are widely used in chewing gum. D-Tagatose is also recognized as a tooth-friendly sweetener and is not fermented by cariogenic bacteria in dental plaque. Consistent with previous studies, D-tagatose has been shown to be a non-fermentable sugar for dental plaque bacteria. S. mutans GS-5, and gas chromatography-mass spectrometry analysis results revealed that 81.6% of the D-tagatose added to the culture media was retained, even after 48 h of S. mutans GS-5 (data not shown). Although the addition of 1% D-tagatose to the culture media retarded the growth of S. mutans GS-5, the final growth yield did not change compared to sugar-free cultures. However, D-tagatose inhibited biofilm formation of S.mutans GS-5, indicating that the effect is caused by a mechanism other than growth inhibition. As demonstrated, D-tagatose inhibits cell-associated GTF activities, resulting in reduced D-fructose release from sucrose. D-fructose (and sucrose) appears to be a potent inducer of gtfB expression. The addition of 1% sucrose to the culture medium induced the expression of the D-fructose-specific PTS and gtfB, indicating that the glucan production and energy metabolism pathways that utilize D-fructose are tightly coordinated in S. mutans. This result is consistent with the findings of Shemesh et al., who demonstrated that D-fructose induces higher levels of gtfB expression than D-glucose in the early exponential phase. Therefore, suppression of gtfB D-tagatose expression may be partially caused by a decrease in D-fructose intake.Furthermore, the genes encoding the EII component for the D-fructose-specific PTS genes (ptsfru and ptsfru / man) were also downregulated in the presence of D-tagatose. The growth retardation of S. mutans GS-5 by D-tagatose may also be due to the limited D-fructose supply resulting from GTF inhibition, since the growth retardation induced by D-tagatose was reversed by D-fructose supplementation. It is therefore hypothesized that alterations in the availability of this monosaccharide are responsible for the prolongation of the growth retardation phase of S. mutans GS-5 by D-tagatose. In contrast, ftf expression levels were not affected by D-tagatose in the presence of 1% sucrose. FTF produces water-soluble inulin-type fructans in S. mutans. Because fructose is digested by FruA to Dfructose, this fructose polymer is thought to serve as an energy depot for S. mutans.Downregulation of fruA by D-tagatose is hypothesized to limit D-fructose supply to S. mutans. This alteration in monosaccharide availability could affect the expression of fruA, which is known to be sensitive to the control of carbon catabolite repression via the central regulatory protein CcpA. As mentioned, D-tagatose appears to inhibit GTFs in S. mutans GS-5, and inhibition of cell-associated GTFs B or C, which produce water-insoluble glucan from sucrose, is considered a primary mechanism underlying D-tagatose-induced biofilm inhibition and growth retardation in S. mutans GS-5. Because D-tagatose is an epimer of D-fructose at the C-4 position, its structural similarity to D-fructose could interfere with the binding or catalysis of sucrose by GTFs. Furthermore, S. mutans biofilms are not affected by D-tagatose.mutans GS-5 formed in the presence of D-tagatose, whereas the biofilm formed in the culture with sucrose alone or sucrose plus xylitol was uniform. This difference may be related to the imbalance between glucans and fructans caused by D-tagatose. Bautista et al. reported that many human pathogens are unable to utilize D-tagatose and demonstrated that the sugar is metabolized by a limited number of lactobacilli. Probiotic lactobacilli have been reported to suppress the growth of cariogenic bacteria and prevent dental caries. Based on the result of the present study, which showed that S. mutans GS-5 does not preferentially ferment D-tagatose, it is expected that this sugar prevents S. mutans colonization on tooth surfaces by promoting the ability of probiotic oral lactobacilli to resist colonization.Xylitol is widely used for the prevention of dental caries, although its effects in clinical studies remain controversial. S. mutans transports xylitol through a fructose-specific PTS, and xylitol-resistant S. mutans strains lacking this PTS activity have emerged. Furthermore, the presence of fermentable sugars, such as sucrose, attenuates the effects of xylitol. Therefore, alternative prophylactic treatments for dental caries are needed. Xylitol is a non-fermentable sugar for S. mutans and exerts a toxic effect by requiring energy expenditure for the absorption and export of this non-cariogenic sugar alcohol. The mechanism by which xylitol suppresses S. mutans appears to be different from that of D-tagatose described here; therefore, a synergistic effect might be expected when combined.However, a synergistic effect was not evident in inhibiting S. mutans GS-5 biofilm formation, which may be related to interference with xylitol absorption, since D-tagatose downregulates the D-fructose-specific PTS genes ptsfru and ptsfru / man. In conclusion, D-tagatose appears to inhibit S. mutans GS-5 growth and biofilm formation by interfering with GTF activity. This effect may be beneficial in preventing dental caries. Based on the results obtained in the present study, foods or preparations containing D-tagatose could be useful tools for improving oral hygiene. D-tagatose may be able to suppress the intermittent growth of S. mutans between oral hygiene activities. Furthermore, S. mutans produces a granular biofilm in the presence of D-tagatose, which may facilitate biofilm removal by mechanical brushing compared to homogeneous biofilms. In light of the bibliographic data and experimental evidence, we can support and confirm a synergistic effect of efficacy between a trace element such as copper and a rare sugar D-Tagatose both in terms of duration over time and efficacy on biofilm-producing pathogens, the golden standard of which is represented by Streptococcus mutans. 7. Composition examples Mouthwash D. Tagatose................................................................. gr10 Colloidal copper................................................ppm40 Copper chlorophyll................................................................gr1 Demineralized water.................................................as much as 100 ml Oral topical gel (w / w) bio-ecological composition. 90% DEMINERALISED WATER BIOMER 0.6% D-TAGATOSE 9.3% COLLOIDAL COPPER 0.1% Oral topical gel (w / w) D. Tagatose.........................................................................gr 10 Colloidal copper................................................................ppm40 Copper chlorophyll................................................................gr1 Carbopol 940........................................................................................gr2.5 Propylene glycol........................................................................................ gr10 Demineralized water qba................................................................100g 8. Conclusion For the intended use in the human mouth and pharynx, an oral composition must have a greater germicidal effect than known oral compositions, with a high degree of tolerance for humans or human tissues and with the exclusion of the possible formation of resistance. The risk / benefit ratio presents a clear improvement over the state of the art, as required by the code of ethics and the regulatory guidelines. To this end, according to the invention, an oral composition is provided, which comprises as components a sugar and a trace element. Preferably, as the composition active on the biofilm, a combination of copper and D-Tagatose is provided. 8.1. In light of what has been masterfully expressed Cases in which an inventive step can be presumed to exist are those in which the invention achieves: an unexpected or surprising effect; a synergistic effect generated by the combination of known elements; a practical solution to a problem that has remained unsolved for a long time, even if the proposed solution appears to be rather simple after the inventor has described it in the patent; a solution in which a technical prejudice is overcome. From CINQUANTINI B., PRIMICERI MV, La proprietà intelligente ei brevetti - guida pratica, Rome, 2015, p. 19. This application is considered worthy of a patent.

Claims

1) Microbiologically active composition comprising at least one or more trace elements of groups 10, 11 and 12 according to the Mendeleev periodic table in elemental form and / or its salts and / or derivatives and a sugar 2) Microbiologically active composition as per the application, wherein such elements are preferably colloidal copper and D-Tagatose 3) Microbiologically active composition according to the previous statements, wherein said elements have a proportional concentration between them of between 0.001% and 99.999% for each single element in any physical form, for example liquid, solid, gaseous, aerosol, spray, nebulised and combinations thereof 4) Microbiologically active composition according to the previous statements, in which said elements have a proportional concentration between them of between 0.001% and 99.999% for each single element in any physical form, for example liquid, solid, gaseous, aerosol, spray, nebulized and combinations thereof, dissolved or incorporated in excipients suitable for the desired formulations: gels, mouthwashes, toothpastes, washes and any device, including medical devices, for use on mucous tissue and dermis. 5) Microbiologically active composition according to the previous indications, including: D.Tagatose..................................................................................10g Colloidal copper..................................................................40ppm Copper chlorophyll..................................................................1g Demineralized water..................................................................as much as 100ml 6) Microbiologically active composition according to the previous indications including DEMINERALIZED WATER 90% BIOMER..................................................0.6% D-TAGATOSE.......................................9.6% COLLOIDAL COPPER...........................0.1% 7) Microbiologically active composition according to the previous indications including: D.Tagatose..................................................................................10g Copper >100 nm.............................................................40ppm Copper chlorophyll..................................................................1g Carbopol 940...........................................................2.5g Propylene glycol...........................................10g Purified water qba........................100g 8) Microbiologically active composition according to the previous indications for use against any microbiological pathogens with activity on human mucous membranes and dermis. 9) Microbiologically active composition according to the previous indications for use against any microbiological pathogen for veterinary use. 10) Microbiologically active composition according to the previous indications for use against any microbiological pathogen with activity on medical devices and / or personal protective equipment (PPE) and / or surfaces and / or environments.