Antimicrobials, Resistance anttimicrobial Chemotherapy. Field, D. Google Scholar Muscle building for beginners. CLO 2 may act to disrupt this membrane by peroxidizing the unsaturated fatty acids found in the bilayer lipid membrane. Download PDF.

Non-toxic antimicrobial agents -

They found that the sugars from one sample nearly killed an entire strep colony. In another sample, the sugars were moderately effective while the remaining three samples exhibited a lower level of activity.

In a follow-up study, they are testing more than two dozen additional samples. So far, two broke down the bacterial biofilms and killed the bacteria, four broke down the biofilms but did not kill the bacteria and two killed the bacteria without breaking down the biofilms.

The research was supported by the Department of Veterans Affairs grant CDA-2 1IK2BX, National Institutes of Health grants T32A and 2T32HDA2, National Center for Research Resources grant UL1 RR, National Center for Advancing Translational Sciences grant 2 UL1 TR and the Vanderbilt Institute of Chemical Biology.

Vanderbilt University. Close Menu Vanderbilt University. Research News All Vanderbilt. Time-kill studies demonstrated that the analog [Lys3, Lys10, Lys14, Lys21]pseudin-2 at a concentration of 1 x MIC was bacteriocidal against E.

coli Increasing the hydrophobicity of pseudin-2, while maintaining the amphipathic character of the molecule, by substitution of neutral amino acids on the hydrophobic face of the alpha-helix by L-phenylalanine, had only minor effects on antimicrobial and hemolytic activities.

Alcohols have been used effectively to disinfect oral and rectal thermometers , , hospital pagers , scissors , and stethoscopes Alcohols have been used to disinfect fiberoptic endoscopes , but failure of this disinfectant have lead to infection , Alcohol towelettes have been used for years to disinfect small surfaces such as rubber stoppers of multiple-dose medication vials or vaccine bottles.

Furthermore, alcohol occasionally is used to disinfect external surfaces of equipment e. In contrast, three bloodstream infection outbreaks have been described when alcohol was used to disinfect transducer heads in an intensive-care setting The documented shortcomings of alcohols on equipment are that they damage the shellac mountings of lensed instruments, tend to swell and harden rubber and certain plastic tubing after prolonged and repeated use, bleach rubber and plastic tiles and damage tonometer tips by deterioration of the glue after the equivalent of 1 working year of routine use Tonometer biprisms soaked in alcohol for 4 days developed rough front surfaces that potentially could cause corneal damage; this appeared to be caused by weakening of the cementing substances used to fabricate the biprisms Corneal opacification has been reported when tonometer tips were swabbed with alcohol immediately before measurement of intraocular pressure Alcohols are flammable and consequently must be stored in a cool, well-ventilated area.

They also evaporate rapidly, making extended exposure time difficult to achieve unless the items are immersed. Hypochlorites, the most widely used of the chlorine disinfectants, are available as liquid e.

The most prevalent chlorine products in the United States are aqueous solutions of 5. They have a broad spectrum of antimicrobial activity, do not leave toxic residues, are unaffected by water hardness, are inexpensive and fast acting , remove dried or fixed organisms and biofilms from surfaces , and have a low incidence of serious toxicity Sodium hypochlorite at the concentration used in household bleach 5.

The microbicidal activity of chlorine is attributed largely to undissociated hypochlorous acid HOCl. A potential hazard is production of the carcinogen bis chloromethyl ether when hypochlorite solutions contact formaldehyde and the production of the animal carcinogen trihalomethane when hot water is hyperchlorinated After reviewing environmental fate and ecologic data, EPA has determined the currently registered uses of hypochlorites will not result in unreasonable adverse effects to the environment Alternative compounds that release chlorine and are used in the health-care setting include demand-release chlorine dioxide, sodium dichloroisocyanurate, and chloramine-T.

The advantage of these compounds over the hypochlorites is that they retain chlorine longer and so exert a more prolonged bactericidal effect. Sodium dichloroisocyanurate tablets are stable, and for two reasons, the microbicidal activity of solutions prepared from sodium dichloroisocyanurate tablets might be greater than that of sodium hypochlorite solutions containing the same total available chlorine.

Second, solutions of sodium dichloroisocyanurate are acidic, whereas sodium hypochlorite solutions are alkaline, and the more microbicidal type of chlorine HOCl is believed to predominate Chlorine dioxide-based disinfectants are prepared fresh as required by mixing the two components base solution [citric acid with preservatives and corrosion inhibitors] and the activator solution [sodium chlorite].

In vitro suspension tests showed that solutions containing about ppm chlorine dioxide achieved a reduction factor exceeding 10 6 of S. aureus in 1 minute and of Bacillus atrophaeus spores in 2. The potential for damaging equipment requires consideration because long-term use can damage the outer plastic coat of the insertion tube In another study, chlorine dioxide solutions at either ppm or 30 ppm killed Mycobacterium avium-intracellulare within 60 seconds after contact but contamination by organic material significantly affected the microbicidal properties The main products of this water are hypochlorous acid e.

As with any germicide, the antimicrobial activity of superoxidized water is strongly affected by the concentration of the active ingredient available free chlorine One manufacturer generates the disinfectant at the point of use by passing a saline solution over coated titanium electrodes at 9 amps.

The product generated has a pH of 5. Although superoxidized water is intended to be generated fresh at the point of use, when tested under clean conditions the disinfectant was effective within 5 minutes when 48 hours old Unfortunately, the equipment required to produce the product can be expensive because parameters such as pH, current, and redox potential must be closely monitored.

The solution is nontoxic to biologic tissues. Although the United Kingdom manufacturer claims the solution is noncorrosive and nondamaging to endoscopes and processing equipment, one flexible endoscope manufacturer Olympus Key-Med, United Kingdom has voided the warranty on the endoscopes if superoxidized water is used to disinfect them As with any germicide formulation, the user should check with the device manufacturer for compatibility with the germicide.

Additional studies are needed to determine whether this solution could be used as an alternative to other disinfectants or antiseptics for hand washing, skin antisepsis, room cleaning, or equipment disinfection e.

In October , the FDA cleared superoxidized water as a high-level disinfectant FDA, personal communication, September 18, The exact mechanism by which free chlorine destroys microorganisms has not been elucidated.

Inactivation by chlorine can result from a number of factors: oxidation of sulfhydryl enzymes and amino acids; ring chlorination of amino acids; loss of intracellular contents; decreased uptake of nutrients; inhibition of protein synthesis; decreased oxygen uptake; oxidation of respiratory components; decreased adenosine triphosphate production; breaks in DNA; and depressed DNA synthesis , The actual microbicidal mechanism of chlorine might involve a combination of these factors or the effect of chlorine on critical sites Low concentrations of free available chlorine e.

Higher concentrations 1, ppm of chlorine are required to kill M. tuberculosis using the Association of Official Analytical Chemists AOAC tuberculocidal test One study reported that 25 different viruses were inactivated in 10 minutes with ppm available chlorine Several studies have demonstrated the effectiveness of diluted sodium hypochlorite and other disinfectants to inactivate HIV Chlorine ppm showed inhibition of Candida after 30 seconds of exposure In experiments using the AOAC Use-Dilution Method, ppm of free chlorine killed 10 6 —10 7 S.

aureus , Salmonella choleraesuis , and P. Because household bleach contains 5. A chlorine dioxide generator has been shown effective for decontaminating flexible endoscopes but it is not currently FDA-cleared for use as a high-level disinfectant Chlorine dioxide can be produced by mixing solutions, such as a solution of chlorine with a solution of sodium chlorite In , a chlorine dioxide product was voluntarily removed from the market when its use caused leakage of cellulose-based dialyzer membranes, which allowed bacteria to migrate from the dialysis fluid side of the dialyzer to the blood side tuberculosis , M.

chelonae , poliovirus, HIV, multidrug-resistant S. aureus , E. coli, Candida albicans , Enterococcus faecalis, P.

aeruginosa in the absence of organic loading. However, the biocidal activity of this disinfectant decreased substantially in the presence of organic material e. No bacteria or viruses were detected on artificially contaminated endoscopes after a 5-minute exposure to superoxidized water and HBV-DNA was not detected from any endoscope experimentally contaminated with HBV-positive mixed sera after a disinfectant exposure time of 7 minutes Hypochlorites are widely used in healthcare facilities in a variety of settings.

A — dilution of 5. For small spills of blood i. Because hypochlorites and other germicides are substantially inactivated in the presence of blood 63, , , , large spills of blood require that the surface be cleaned before an EPA-registered disinfectant or a final concentration solution of household bleach is applied If a sharps injury is possible, the surface initially should be decontaminated 69, , then cleaned and disinfected final concentration Extreme care always should be taken to prevent percutaneous injury.

At least ppm available chlorine for 10 minutes is recommended for decontaminating CPR training manikins Full-strength bleach has been recommended for self-disinfection of needles and syringes used for illicit-drug injection when needle-exchange programs are not available.

The difference in the recommended concentrations of bleach reflects the difficulty of cleaning the interior of needles and syringes and the use of needles and syringes for parenteral injection Clinicians should not alter their use of chlorine on environmental surfaces on the basis of testing methodologies that do not simulate actual disinfection practices , Other uses in healthcare include as an irrigating agent in endodontic treatment and as a disinfectant for manikins, laundry, dental appliances, hydrotherapy tanks 23, 41 , regulated medical waste before disposal , and the water distribution system in hemodialysis centers and hemodialysis machines Chlorine long has been used as the disinfectant in water treatment.

Water disinfection with monochloramine by municipal water-treatment plants substantially reduced the risk for healthcare—associated Legionnaires disease , Chlorine dioxide also has been used to control Legionella in a hospital water supply.

Thus, if a user wished to have a solution containing ppm of available chlorine at day 30, he or she should prepare a solution containing 1, ppm of chlorine at time 0.

Sodium hypochlorite solution does not decompose after 30 days when stored in a closed brown bottle The use of powders, composed of a mixture of a chlorine-releasing agent with highly absorbent resin, for disinfecting spills of body fluids has been evaluated by laboratory tests and hospital ward trials.

The inclusion of acrylic resin particles in formulations markedly increases the volume of fluid that can be soaked up because the resin can absorb — times its own weight of fluid, depending on the fluid consistency.

One problem with chlorine-releasing granules is that they can generate chlorine fumes when applied to urine Formaldehyde is used as a disinfectant and sterilant in both its liquid and gaseous states.

Liquid formaldehyde will be considered briefly in this section, and the gaseous form is reviewed elsewhere The aqueous solution is a bactericide, tuberculocide, fungicide, virucide and sporicide 72, 82, OSHA indicated that formaldehyde should be handled in the workplace as a potential carcinogen and set an employee exposure standard for formaldehyde that limits an 8-hour time-weighted average exposure concentration of 0.

The standard includes a second permissible exposure limit in the form of a short-term exposure limit STEL of 2 ppm that is the maximum exposure allowed during a minute period Ingestion of formaldehyde can be fatal, and long-term exposure to low levels in the air or on the skin can cause asthma-like respiratory problems and skin irritation, such as dermatitis and itching.

For these reasons, employees should have limited direct contact with formaldehyde, and these considerations limit its role in sterilization and disinfection processes. Key provisions of the OSHA standard that protects workers from exposure to formaldehyde appear in Title 29 of the Code of Federal Regulations CFR Part Formaldehyde inactivates microorganisms by alkylating the amino and sulfhydral groups of proteins and ring nitrogen atoms of purine bases Varying concentrations of aqueous formaldehyde solutions destroy a wide range of microorganisms.

Four percent formaldehyde is a tuberculocidal agent, inactivating 10 4 M. tuberculosis in 2 minutes 82 , and 2. anthracis The formaldehyde solution required 2 hours of contact to achieve an inactivation factor of 10 4 , whereas glutaraldehyde required only 15 minutes.

For these reasons and others—such as its role as a suspected human carcinogen linked to nasal cancer and lung cancer , this germicide is excluded from Table 1. When it is used, , direct exposure to employees generally is limited; however, excessive exposures to formaldehyde have been documented for employees of renal transplant units , , and students in a gross anatomy laboratory Formaldehyde is used in the health-care setting to prepare viral vaccines e.

To minimize a potential health hazard to dialysis patients, the dialysis equipment must be thoroughly rinsed and tested for residual formaldehyde before use. Paraformaldehyde, a solid polymer of formaldehyde, can be vaporized by heat for the gaseous decontamination of laminar flow biologic safety cabinets when maintenance work or filter changes require access to the sealed portion of the cabinet.

Glutaraldehyde is a saturated dialdehyde that has gained wide acceptance as a high-level disinfectant and chemical sterilant Aqueous solutions of glutaraldehyde are acidic and generally in this state are not sporicidal. Once activated, these solutions have a shelf-life of minimally 14 days because of the polymerization of the glutaraldehyde molecules at alkaline pH levels.

This polymerization blocks the active sites aldehyde groups of the glutaraldehyde molecules that are responsible for its biocidal activity.

Novel glutaraldehyde formulations e. However, antimicrobial activity depends not only on age but also on use conditions, such as dilution and organic stress. However, two studies found no difference in the microbicidal activity of alkaline and acid glutaraldehydes 73, The biocidal activity of glutaraldehyde results from its alkylation of sulfhydryl, hydroxyl, carboxyl, and amino groups of microorganisms, which alters RNA, DNA, and protein synthesis.

The mechanism of action of glutaraldehydes are reviewed extensively elsewhere , The in vitro inactivation of microorganisms by glutaraldehydes has been extensively investigated and reviewed , Spores of C. Microorganisms with substantial resistance to glutaraldehyde have been reported, including some mycobacteria M.

chelonae , Mycobacterium avium-intracellulare, M. xenopi , Methylobacterium mesophilicum , Trichosporon , fungal ascospores e. chelonae persisted in a 0. Two percent alkaline glutaraldehyde solution inactivated 10 5 M.

tuberculosis cells on the surface of penicylinders within 5 minutes at 18°C However, subsequent studies 82 questioned the mycobactericidal prowess of glutaraldehydes. tuberculosis and compares unfavorably with alcohols, formaldehydes, iodine, and phenol Suspensions of M. avium, M.

FDA antimicrobia, not cleared any liquid Muscle building for beginners Amazon Sports Equipment or high-level disinfectant Non-toxic antimicrobial agents alcohol as the main active ingredient. These alcohols are Non-toxic antimicrobial agents bactericidal rather Non-todic bacteriostatic against vegetative Non-todic of NNon-toxic they Noon-toxic are tuberculocidal, fungicidal, and virucidal but do not destroy bacterial spores. Top of Page. The most feasible explanation for the antimicrobial action of alcohol is denaturation of proteins. This mechanism is supported by the observation that absolute ethyl alcohol, a dehydrating agent, is less bactericidal than mixtures of alcohol and water because proteins are denatured more quickly in the presence of water ,

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Antibiotics Cooling down after workouts world agfnts facing a wntimicrobial increase in infections caused Herbal tea blends drug-resistant infectious agents. In response, Nn-toxic strategies have been recently explored to treat them, anti,icrobial the aantimicrobial of bacteriocins. Bacteriocins are a group of Non-toxic antimicrobial agents peptides Herbal tea blends Noon-toxic bacteria, capable of controlling clinically relevant susceptible and drug-resistant bacteria. Bacteriocins have been studied to be able to modify and improve their physicochemical properties, pharmacological effects, and biosafety. This manuscript focuses on the research being developed on the biosafety of bacteriocins, which is a topic that has not been addressed extensively in previous reviews. This work discusses the studies that have tested the effect of bacteriocins against pathogens and assess their toxicity using in vivo models, including murine and other alternative animal models.

Non-toxic antimicrobial agents -

CLO 2 in pure form anhydrous is a liquid at O°C. Having a deep red color. When mixed with water and at high dilution it is colourless. In the covalent bond one electron is contributed by each of the atoms forming the bond Figure 1.

When CLO 2 reacts as an anti-microbial agent the oxygen atom first binds to a single atom the one being oxidized and then is dissociated from chlorine. An electron is then given up to chlorine forming the chloride ion.

When one realizes that there are 5. of chloride ion per liter of human plasma it becomes obvious that the small amount of chloride generated through the use of CLO 2 is negligible. CLO 2 is an inorganic containing no carbon sodium salt of only electronegative elements bound together by electrostatic; covalent and coordinate covalent bonds Figure 2.

Under acidic conditions; sodium chlorite becomes unstable and decomposes into a variety of products including but not limited to chloride ions; hyperchlorous ions and nascent atomic oxygen.

It is for this reason that CLO 2 when bottled and stored for long periods of time; is buffered. Under acidic conditions; CLO 2 results in a neutral molecule consisting of three electronegative atoms held together by covalent and coordinate covalent bonds. From this cluster; a single atom of highly reactive nascent oxygen is liberated onto the target microorganisms.

As you will see below; nascent oxygen is the active agent of CLO 2 and; unless it is liberated; the antiviral; antibacterial and antifungal activity will not occur.

A virus typically consists of an outer shell or coating of protein encapsulating a nucleic acid which may be either DNA or RNA e. The skeletal backbone of nucleic acids includes derivatives of phosphoric acid H 3 PO 4 ; a very strong acid in which two of the original three hydroxyl groups -OH are substituted; leaving only one active hydroxyl group per phosphate.

Segment of extended dna chain showing alternating deoxyribose- phosphate linkage. Some viruses may have glycoproteins incorporated into their protein coat; that is; proteins to which polysaccharides sugar chains have been attached.

The bound polysaccharides may attach to specific sites on the protein coat; effectively converting a protein surface into a poly- sugar surface.

Once bound to the appropriate cell type; the nucleic acid component of the virus is injected into the cell and in many ways; takes over the protein synthesis processes of the cell. Certain segments of the viral nucleic acid consist of genes that are responsible for the replication of the coat.

The result is the replication of the complete virus until the cell bursts; releasing many additional viral particles into the surrounding medium.

In the presence of these acidic nucleic acids; the CLO 2 molecule becomes unstable and releases nascent oxygen into the medium. Nucleic acids; both RNA and DNA; have many characteristics in common; in fact; they are almost chemically identical. Altogether; there are only four types of bases found in DNA; namely; guanine G ; cytosine C ; adenine A and thymine T.

It is the sequence of these four units along the chain that makes one segment of DNA differs from another. RNA differs chemically from DNA in that the base thymine is replaced by the base uracil U ; representing a very subtle biochemical difference.

There is also a subtle difference in the sugar deoxyribose. The release of nascent oxygen from chlorine dioxide CLO 2 in an acidic environment. The base guanine; found in both RNA and DNA; is very sensitive to oxidation; forming 8-oxoguanine as the oxidation product.

Although the replication of the protein coat may continue; the formation of a complete functional virus has been blocked by CLO 2 oxidation. These bacteria survive only in the absence of oxygen and include many pathogenic organisms.

With the appearance of algae; carbon dioxide; present in large proportion in the primordial atmosphere; was converted by algae to carbohydrates; generating oxygen as a by product.

At first; oxygen reacted with iron salts in the primitive oceans to generate extensive deposits of iron oxide iron ore. When soluble iron in the oceans became exhausted; oxygen concentration in the atmosphere began to rise.

Some bacteria were able to survive the increased levels of oxygen in the atmosphere by any of three paths. The presence of oxygen in the atmosphere resulting from algae and other plants containing the green pigment chlorophyll; allowed the development of the wide range of life forms both in the ocean and on land known today as animals.

Some anaerobic bacteria took up residence in oxygen-free environments within the bodies of animals; notably; the intestines. Other anaerobes solved the problem of oxygen toxicity by evolving metabolic ways to cope with its universal presence. These include the development of the enzyme superoxide dismutase SOD ; which in its most primitive form; contained iron.

It is widely believed that still another solution to oxygen toxicity lay in the development of organisms that have the ability to utilize oxygen in their metabolism. These organisms are known today as the aerobes or aerobic bacteria.

One form of these organisms may have been the precursor of an inclusion organism found in many animal and human cells known as the mitochondria; having its own circular DNA. It is well known that cultures of many bacteria become acidic; typically generating lactic; acetic and other simple carboxylic organic acids.

The acidic medium surrounding many bacteria triggers the decomposition of CLO 2 and the subsequent liberation of nascent oxygen. Nascent oxygen is a particularly potent oxidizing agent for anaerobic organisms because it is essentially a free radical seeking not one; but two electrons.

Anaerobic organisms have not developed adequate defences against the onslaught of oxygen; particularly nascent oxygen; and quickly succumb to its lethal action.

This association is equivalent to the formation of hydrogen — chlorine H — Cl covalent bond. Because of the single electron deficiency of the chlorine atom; only one covalent bond is permitted at any one time. Since the H — Cl bond is stronger the H — O bond of water is almost the strongest bond known than the existing O — Cl covalent bond; the O — Cl bond is disrupted by the ejection of nascent oxygen.

This leads to the dissociation of the acidic proton from chlorine and the formation of the hypochlorous ion ClO- and sodium hypochlorite NaOCl and the release of nascent oxygen from chlorine dioxide in an acidic environment.

The decomposition of sodium chlorite to nascent oxygen and sodium hypochlorite in an acidic environment. Figure 3 The formula for decomposition of sodium chlorite to nascent oxygen and sodium hypochlorite in an acidic environment].

During World War I; this instability of chlorite ion was exploited by Alexis Carrel —; Nobel laureate ; best known for his prolonged culture of chicken heart cells at Rockefeller University; New York.

Carrel sucessfully administered a crude solution topically to war casualties having embedded shell fragments. Figure 3 An electron micrograph showing the intact polio virus before exposure to ozone O 3.

There are only a small number of basic biological substances found in living organisms. These include proteins; lipids; carbohydrates and nucleic acids. The ability of nucleic acids in the form of viruses to activate chlorine dioxide and then nascent oxygen has been discussed previously.

Lipids and carbohydrates are both neutral substances; carrying neither an electric charge nor acidic groups. The possibility of proteins activating CLO 2 is discussed below. Many proteins; particularly those that are soluble and found free in both the blood and in the medium surrounding tissues; contain on their surfaces organic groups; both acidic and basic alkaline in nature.

In most soluble globular proteins either one or the other of these two types of groups predominate. Proteins in which the acidic groups outweigh the basic groups are acidic proteins and those in which the basic groups predominate are known as basic proteins.

A technique has been developed to determine; for a given protein; in which category it may lie. Either acidic or basic proteins migrate under the influence of an applied electric field. This laboratory technique; known as electrophoresis; has been used to determine the acidity or basicity of proteins.

As the pH of the surrounding medium into which a protein has been solubilized is changed; the protein will migrate either in one direction or the other. When a pH is found that will result in no migration; the protein is said to be at its isoelectric point. The pH value at which this occurs for a given protein is known as the isoelectric point of that protein.

The isoelectric points of most proteins are not far from 7 or neutrality. Such proteins are not capable of activating CLO 2 to decomposition; partly accounting for low toxicity.

Fungi are considered as plants without chlorophyll and lack the ability to generate carbohydrates from sunlight and carbon dioxide. They were probably derived in an oxygen-free atmosphere but some have developed the ability to tolerate low levels of oxygen.

Most fungi prefer an oxygen-poor environment and live best under these conditions. They obtain their energy requirements from the decomposition through enzyme activity of existing organic matter and; in this light; may be considered as parasitic.

Because of their low tolerance for nascent oxygen and the acidic medium in which they thrive from the liberation of organic acids ; fungi in the mycelial form are sensitive to the destructive action of CLO 2.

One example of a human pathogenic fungus is Candida albicans; invading finger and toenails. From all of the above considerations; it is apparent that CLO 2 is essentially activated only by viruses; acidic bacteria and fungi. Other Cytotoxic Oxidizing Agents Used Clinically CL02 is not the only anti-microbial agent in clinical use.

Another agent also providing active oxygen is hydrogen peroxide; which has been used in the treatment of arthritis; cancer and other metabolic diseases. Hydrogen peroxide is commercially available in low concentrations for the treatment of topical microbial infections.

Found to be more effective as a sporicide than as a bactericide; hydrogen peroxide is bacteriostatic at concentrations greater than 0. This bonding is unstable; decomposing to acetic acid and one atom of oxygen is being made available as bactericidal; fungicidal or virucidal. Peracetic acid exhibits rapid activity against spores and yeasts.

An anti-microbial agent having chemical similarity to CLO 2 is sodium hypochlorite NaOCI in which only one atom of oxygen is bound to halogen rather than two. Solutions containing ppm were found effective within 30minutes in decontaminating objects which had been massively contaminated by both gram-positive and gram-negative bacteria and fungi.

Drinking water heavily contaminated by a polymicrobial suspension became sterile in 30minutes by a solution containing 10ppm of sodium hypochlorite. Still another oxidizing agent having increased application is ozone. Ozone is an unstable gas and is not commercially available but must be generated as required.

It has been shown that ozone selectively inhibits the growth of human cancer cells. These results indicate that cancerous cells; because of their altered oxygen metabolism; are less able to handle the oxidative stress presented by ozone than normal cells.

Ozone has also been shown to inactivate polio virus with an effect noticeable after only 0. These results parallel those caused by CLO 2 in relation to polio virus see below, anti-viral activity of CLO 2.

Ozone is being used both in Europe and the U. Figure 4 Shows a Phase contrast micrograph of the polio virus after exposure to ozone O 3. Proof that CLO 2 is cytotoxic to bacteria; fungus and virus clinically is shown by data indicating its effectiveness as a disinfectant outside the body.

A germicidal solution has been developed containing CLO 2 at an acid PH lactic acid. The solution gave complete kill of Staphylococcus aureus; Pseudomonas and Candida albicans spores within 10minutes. If used in an ultrasound cleaning device complete killing occurred in less than five minutes.

CLO 2 applied to polio virus separated the RNA from the protein coat capsid. CLO 2 reacted with the capsid protein and prevented the adsorption; penetration and normal un-coating of the virus.

It also reacted with the viral RNA and impaired the ability of the nucleic acid to act as a template for replication. The half-life for the elimination of CLO 2 from experimental animals rat was The distribution of CLO2 in tissues following administration of CLO2 labelled material is as follows.

Plasma; 2. Figure 5 Is a scanning electron micrograph of Candida albicans showing the bud scars. It has been shown by toxicity studies in man that daily ingestion of ml CLO 2 having a concentration of 5ppm is safely tolerated.

It has been demonstrated clinically that CLO 2 is highly effective in accelerating wound healing with particular applications to burns. There may at first be no apparent explanation for such a relationship but careful biochemical analysis has revealed a mechanism of action relating CLO 2 to wound healing.

Nucleic acids RNA and DNA are formed of only four basic units linked together into a chain. One of the basic units is designated; guanosine monophosphate; or GMP. If the phosphate group is brought close to one of the hydroxyl groups of the sugar component ribose; water is eliminated and the product is known as cyclic GMP cGMP.

It has been shown that cGMP stimulates cell division and is activated during the regeneration process. It is also known that the enzyme synthesizing cGMP; guanylate cyclase; is stimulated by one of the highly reactive oxygen derivatives; hydroxyl radical; symbolized by OH-.

An oxidizing agent chemically similar to CLO 2 is periodic acid; HIO4; in which four oxygen atoms are bound to a single halogen atom; iodine. In CLO 2 ; two oxygen atoms are bound to the halogen chlorine. It has been determined that hydroxyl radical is present in solutions of periodic acid 24 and; by inference; may also be present in solutions of CLO 2.

The presence of hydroxyl radicals in CLO 2 activates the enzyme guanylate cyclase; which; in turn; synthesizes cGMP leading to cell proliferation and wound healing. Candida albicans is a normal resident of the human digestive system; and vaginal tract and; unlike many other yeasts; is found in two forms dimorphic.

This process is also accompanied by branching in which a new tube may start to grow laterally from a specific point. By changing the culture conditions the form of Candida albicans may be changed from one form to the other and vice versa.

This phenomena is known as pleomorphism Figure 6. Figure 6 Y-form or round Candida albicans can biologically transform into a M-form or mycelial Candida albicans. It is believed that only the mycelial form is pathogenic in man; who will be discussed in greater detail below.

The incidence of both superficial and invasive candidosis has been increased markedly over the last few decades; probably resulting from the widespread usage of immunosuppressive treatment; antibiotics; etc.

The organism can reasonably be described as the most common and most serious fungal pathogen of man. Candida albicans has a worldwide distribution and is commonly found in normal individuals. The incidence of this fungus in various groups undergoing medical treatment is in general higher.

Candida vaginitis is at present the second most common form of vaginal infection in the United States. Of the women who develop vaginal candidosis; a significant proportion suffer frequent recurrences or show chronic intractable symptoms. Candida albicans can infect virtually every tissue in the human body.

Clearly there is a requirement for new and more effective antifungal drugs. For Candida albicans to successfully colonize and infect mucosal surfaces it has to adhere to epithelial surfaces. In germ free animals the mucosa is colonized in higher numbers than in that of control animals normal.

These results suggest that the indigenous bacterial flora suppresses Candida albicans colonization. This; in turn; implies that at least one factor contributing to Candida albicans infection may be the balance between the various components of the intestinal flora.

The lysis of Candida and other infectious yeasts by CLO 2 may be accounted for by a simple mechanism related to the oxidizing ability of this substance. There follows a description and rationale for the mechanism as related to chitin; one of the constituents of the fungal cell wall.

Chitin is a polymeric substance formed by linking together a basic subunit forming the bulk of the hard parts of insects shell; legs; head; etc.

It is a water-insoluble substance formed by linking together the glucose derivative; N-acetylglucosamine NAG contains nitrogen while glucose does not. Chitin is biochemically synthesized by the enzyme chitin synthetase; and provides rigidity to the fungal cell wall.

Yeast cells reproduce asexually by budding in which a daughter cell is formed by a constriction near one end of a mature cell. The construction continues until the walls almost meet followed by a thickening of the wall at this point. The space remaining is filled with a plug of chitin ki-tin.

When the daughter cell breaks free what would otherwise be an open hole in the cell becomes a bud scar of chitin plugging the hole and preventing cell disruption through lysis 28 see Figure 3.

Thus; any agent which inhibits the synthesis of chitin should; in the light of these considerations; act as a lytic agent for yeast. In an effort to answer the question as to whether the cellular immune system white blood cells has an anti-fungal action against Coccidioides immitis; polymorphonuclear leukocytes PMN cells were incubated with the fungus under culture conditions.

The results of contacting PMNs with the fungus indicated that the incorporation of NAG into chitin was inhibited from 54 to 85 percent.

PMNs from patients with chronic granulomatous disease CGD are apparently equivalent in every way to similar cells from normal individuals but lack the ability to generate active oxygen species members of the Reactive Oxygen Toxic Species or ROTS family used by these cells to fight infection including superoxide; hydrogen peroxide; hydroxyl radical and singlet oxygen.

One measure of the production of reactive oxygen toxic species or ROTS by these cells is the chemiluminescence chemical generation of light resulting from the presence of foreign particles.

Tests made with PMNs isolated from patients with CGD are remarkably low in chemiluminescent response to stimulation. When PMN's from a patient with CGD were incubated with Coccidioides; the incorporation of NAG into chitin was inhibited from only 0 to 22 percent under various conditions as compared to percent with normal PMNs.

Similar oxidative species arise from the breakdown of certain oxidizing agents including CLO 2 when in contact with the target substances bacterial proteins; polysaccharides; viruses; etc. It has been shown that periodic acid HIO4 ; a substance in which four oxygen atoms are bound to iodine corresponding to chlorine in CLO 2 ; is capable of forming one of the ROTS family; hydroxyl radical.

Another reactive oxygen derivative; superoxide; may result from the breakdown of CLO 2 during an oxidative reaction in which the two oxygen atoms released acquire an electron. It is known to be produced by Candida albicans as a metabolic by-product. These considerations indicate that CLO 2 as well as PMN cells are capable of inhibiting the biosynthesis of chitin in Candida resulting in lysis from the absence of the chitin plug at the bud scar.

Viruses; in general; consist of two parts; an inner nucleic acid core consisting of either; DNA or RNA; and an outer protein coat.

The protein coat not only serves to protect the vital nucleic acid within but is also instrumental in providing specificity in binding to the surface of particular cells polio virus binds to nerve cells; influenza virus binds to the nasal mucosa; etc.

It has also been shown that without certain specific polysaccharides attached to the protein coat glycoproteins ; binding to the cell surface will not occur. This aspect will be discussed in greater detail below in relation to the action of CLO 2 on viruses.

Some virus; including the AIDS virus HTLV-I3I; herpes and cytomegalovirus; are surrounded by a bilayer lipid membrane which is a fragment of the plasma membrane of the cell in which the virus replicated. As the virus ruptures and kills the cell it takes with it a fragment of the plasma membrane of that cell and becomes encapsulated.

This membrane is critical for infectivity and without it the virus cannot enter the proper cell type for replication.

Anti-viral agents which damage or deteriorate the encapsulating bilayer lipid membrane also inhibit or prevent infectivity.

We shall return to this aspect of the anti-viral activity of CLO 2 in a subsequent section. The action of CLO 2 as a cytotoxic agent may include as many as four separate biochemical mechanisms described below. As mentioned above; the protein coat is highly significant in viral infectivity and provides specificity for binding to the proper cell type.

Two amino acids that would undergo oxidation and modification to a significant enough degree to permit inhibition of virus to cell binding are those containing sulfur; namely; methionine and cysteine.

In the first stage the binding of one atom of oxygen results in the formation of the sulfoxide -CH2CH3SCO2CH; having the ability to bind several molecules of water hydrophilic.

The parent compound; methionine; is hydrophobic in relation to the residue. A subsequent stage of oxidation leads to the formation of the sulfone; -CH 2 CH 2 S O 2 CH; which is even more hydrohpilic than the sulfoxide.

Both methionine sulfoxide and sulfone are well known modifications of this amino acid. The modification of the water binding ability of a protein constituent methionine is highly significant in relation to the antiviral activity of CLO 2.

In transmission electron micrographs of virus which has been exposed to the action of CLO 2 it appears that the outer coat has swollen much like a sponge. The increased penetration of water into the viral protein coat allows additional CLO 2 water soluble to penetrate to still deeper layers of the coat; thus initiating a vicious cycle Figure 3.

As mentioned above; CLO 2 is capable of generating a variety of extremely reactive oxygen derivatives when acting as an oxidizing agent. Another example of an oxidizing agent releasing atomic or nascent oxygen is ozone; O 3 this molecule consists of a chain of three oxygen atoms held together by covalent bonds.

The release of one atom of oxygen results in stable; molecular oxygen; 0. The oxidizing activity of CLO 2 is similar to that of ozone; liberating highly active forms of oxygen. Very detailed research has been described related to the oxidation of polio virus by ozone.

Exposure of the virus RNA strand to ozone resulted in fragmentation into several short chains. The effect of ozone on the nucleic acid was measured spectrophotometrically indicating that inactivation was complete after only sixty seconds.

Nucleic acid comprising the RNA or DNA of viruses consists of a chain of four building blocks arranged in some specific sequence. It is the sequence of the four nucleotides designated by the letters A; C; G and U for RNA that causes one strand to differ from another resulting in the synthesis of specific proteins for each.

When a mixture of the four nucleotides is exposed to ozone it is found that only the unit designated; G guanine ; is degraded in the initial stage.

In the exposure of transfer RNA tRNA to ozone; guanine again was the first nucleotide to be degraded. Exposure of tobacco mosaic virus TMV to ozone caused a rapid loss of infectivity within 30minutes showing a preferential degradation of guanine.

Some viruses are surrounded by a bilayer lipid membrane as described above. Any substance which acts to disrupt this membrane is an anti-viral agent for that virus since the membrane is essential for infectivity. CLO 2 may act to disrupt this membrane by peroxidizing the unsaturated fatty acids found in the bilayer lipid membrane.

The specific positioning of these unsaturations along the fatty acid chain is related to the integrity and fluidity of the membrane. The peroxidation of unsaturations results in the development of hydrophilic water seeking groups in the central part of the fatty acid chain which causes distortion in the bilayer lipid membrane.

This activity of CLO 2 has not been demonstrated biochemically but is very apparent in the electron micrographs transmission of viral membrane disruption shown in Figure There are found on the surfaces of viruses; bacteria and fungi including C. albicans chains of sugars polysaccharides ; some attached to protein as glycoprotein.

An example of such a polysaccharide is mannan composed principally of sugar mannose and found on the surface of many yeasts. The degradation of sugars and polysaccharides has been conducted mainly with periodic acid and its salts having the structure HIO 4. The oxidizing part of this substance is the ion IO 4 - periodate ; in which four oxygen atoms are bound to a single atom of iodine halogen.

A similar oxidizing agent is pH Miracle's CLO 2 product called Activator having the structured CLO 2 or in its partially reduced form; CLO. The products formed are low molecular weight substances including formaldehyde; formic acid; carbon dioxide; etc.

The surfaces of many organisms and viruses carry polysaccharides that are highly significant in relation to infectivity. It is these polysaccharides showing specificity for the surface of target cells that enables the organism to bind to those particular cells and invade infectivity.

The principal cell wall polysaccharides of most yeasts are mannans formed mainly of the sugar; mannose and glucans formed mainly of the sugar; glucose whose basic structures have been determined.

In models of the yeast cell wall recently proposed; a mannan-protein component is found in the outer region while glucan is found mainly in the inner region. Chitin a polymer of the glucose derivative; N-acetylglucosamine has been detected not only in the bud scar but also in the glucan network of the cell wall.

Treatment of the cells yeast form of C. albicans with helicase; an enzyme preparation from snail gut; releases mostly mannose from the surface during the first 20minutes. Using selective stains for electron microscopy as many as eight discrete layers have been demonstrated in the cell wall of C.

The outermost layer; 1; consists mostly of 1;6-mannan. The second layer is composed of 1;2-mannan while the third layer is 1;6-glucan. The fourth layer is largely 1;3-glucan with the fifth layer; chitin. The innermost layers; 6;7 and 8; are composed largely of chitin-protein.

These results confirm those described previously employing chemical extraction and enzymic digestion of the individual layers 33 Figure 8. Figure 8 Shows the cell wall following digestion by the enzyme preparation, helicase.

The Y- form reproduces by budding in which an older cell constricts at a point near one end and continues to narrow until a new daughter cell is pinched off. The M-form continues to elongate at the top; laying down a septum as it progresses. A great effort has been made by researchers to determine what factors or set or culture conditions are responsible for the Y to M conversion in C.

albicans and vice versa. No one single causative factor has yet been defined although there is evidence that the factors are complex. Much attention has been given to this subject because there is a large body of literature that claims a relationship between the mycelial M-form and infectivity.

Figure 10 illustrated differences in morphology in spore development at two different temperatures; 20° and 37°C. In growing C.

albicans in a chemically defined medium it has been found that biotin is essential for supporting maximal growth of various strains of this fungus. The biotin level of normal human blood as well as several tissues such as liver and brain is known to be in the range of 0.

For these reasons there is a possibility that this vitamin plays a very definite role in determining the extent and the phase of Candida albicans growth in vivo. When the polysaccharide composition of cell walls isolated from normal-biotin and biotin-deficient cultures was analyzed it was found that the mannan outer layer content of biotin-insufficient cultures was 20 percent less than that of biotin-optimal cultures while the glucan inner layer content was 50 percent or more greater in biotin-deficient cultures.

In other embodiments of the present invention, the compositions further comprise one or more germination enhancing compounds e. In preferred embodiments, the germination enhancing compound is provided in the aqueous phase prior to formation of the emulsion. The present invention contemplates that when germination enhancers are added to the disclosed compositions the sporicidal properties of the compositions are enhanced.

The present invention further contemplates that such germination enhancers initiate sporicidal activity near neutral pH between pH , and preferably 7. Such neutral pH emulsions can be obtained, for example, by diluting with phosphate buffer saline PBS or by preparations of neutral emulsions.

The sporicidal activity of the compositions preferentially occurs when the spores initiate germination. In specific embodiments, it has been demonstrated that the emulsions of the present invention have sporicidal activity. While the present invention is not limited to any particular mechanism, it is believed that the fusigenic component of the emulsions acts to initiate germination and before reversion to the vegetative form is complete the lysogenic component of the emulsion acts to lyse the newly germinating spore.

These components of the emulsion thus act in concert to leave the spore susceptible to disruption by the emulsions. The addition of germination enhancer further facilitates the anti-sporicidal activity of the emulsions of the present invention, for example, by speeding up the rate at which the sporicidal activity occurs.

Germination of bacterial endospores and fungal spores is associated with increased metabolism and decreased resistance to heat and chemical reactants.

For germination to occur, the spore must sense that the environment is adequate to support vegetation and reproduction. The amino acid L-alanine stimulates bacterial spore germination See e. L-alanine and L-proline have also been reported to initiate fungal spore germination Yanagita, Arch Mikrobiol [].

Simple α-amino acids, such as glycine and L-alanine, occupy a central position in metabolism. Transamination or deamination of α-amino acids yields the glycogenic or ketogenic carbohydrates and the nitrogen needed for metabolism and growth. For example, transamination or deamination of L-alanine yields pyruvate which is the end product of glycolytic metabolism Embden-Meyerhof-Parnas Pathway.

Acetyl-CoA is the initiator substrate for the tricarboxylic acid cycle Kreb's Cycle which in turns feeds the mitochondrial electron transport chain. Acetyl-CoA is also the ultimate carbon source for fatty acid synthesis as well as for sterol synthesis.

In certain embodiments, suitable germination enhancing agents of the invention include, but are not limited to, α-amino acids comprising glycine and the L-enantiomers of alanine, valine, leucine, isoleucine, serine, threonine, lysine, phenylalanine, tyrosine, and the alkyl esters thereof.

Additional information on the effects of amino acids on germination may be found in U. In some embodiments, a mixture of glucose, fructose, asparagine, sodium chloride NaCl , ammonium chloride NH 4 Cl , calcium chloride CaCl 2 and potassium chloride KCl also may be used.

In particularly preferred embodiments of the present invention, the formulation comprises the germination enhancers L-alanine, CaCl 2 , Inosine and NH 4 Cl. In some embodiments, the compositions further comprise one or more common forms of growth media e.

The above compounds are merely exemplary germination enhancers and it is understood that other known germination enhancers will find use in the compositions of the present invention.

A candidate germination enhancer should meet two criteria for inclusion in the compositions of the present invention: it should be capable of being associated with the emulsions of the present invention and it should increase the rate of germination of a target spore in the when incorporated in the emulsions of the present invention.

One skilled in the art can determine whether a particular agent has the desired function of acting as an germination enhancer by applying such an agent in combination with the compositions of the present invention to a target and comparing the inactivation of the target when contacted by the admixture with inactivation of like targets by the composition of the present invention without the agent.

Any agent that increases germination, and thereby decrease or inhibits the growth of the organisms, is considered a suitable enhancer for use in the present invention. In still other embodiments, addition of a germination enhancer or growth medium to a neutral emulsion composition produces a composition that is useful in treating bacterial spores in addition to enveloped viruses, Gram negative bacteria, and Gram positive bacteria.

In still other embodiments, the compositions of the present invention comprise one or more compounds capable of increasing the interaction of the compositions i. In preferred embodiments, the interaction enhancer is preferably premixed with the oil phase; however, in other embodiments the interaction enhancer is provided in combination with the compositions after emulsification.

In certain preferred embodiments, the interaction enhancer is a chelating agent e. It is understood that chelating agents are merely exemplary interaction enhancing compounds. In particularly preferred embodiments, the interaction enhancer is at a concentration of about 50 to about μM, although higher and lower amounts are contemplated.

One skilled in the art will be able to determine whether a particular agent has the desired function of acting as an interaction enhancer by applying such an agent in combination with the compositions of the present invention to a target and comparing the inactivation of the target when contacted by the admixture with inactivation of like targets by the composition of the present invention without the agent.

Any agent that increases the interaction and thereby decrease or inhibits the growth of the bacteria in comparison to that parameter in its absence is considered an interaction enhancer. In some embodiments, the addition of an interaction enhancer to the compositions of the present invention produces a composition that is useful in treating enveloped viruses, some Gram positive bacteria and some Gram negative bacteria.

In some embodiments of the present invention, the nanoemulsion composition comprise one or more additional components to provide a desired property or functionality to the nanoemulsions. These components may be incorporated into the aqueous phase or the oil phase of the nanoemulsions and may be added prior to or following emulsification.

For example, in some embodiments, the nanoemulsions further comprise phenols e. In section A , set forth below, the present invention describes exemplary techniques for making generic formulations of the disclosed compositions. Additionally, the present invention recites a number of specific, although exemplary, formulation recipes in section B set forth below.

The pathogen inactivating oil-in-water emulsions of the present invention can be formed using classic emulsion forming techniques.

In brief, the oil phase is mixed with the aqueous phase under relatively high shear forces e. The emulsion is formed by blending the oil phase with an aqueous phase on a volume-to-volume basis ranging from about to , preferably about to , most preferably , oil phase to aqueous phase.

The oil and aqueous phases can be blended using any apparatus capable of producing shear forces sufficient to form an emulsion such as French Presses or high shear mixers e. Methods of producing such emulsions are described in U. In preferred embodiments, the compositions used in the methods of the present invention comprise droplets of an oily discontinuous phase dispersed in an aqueous continuous phase, such as water.

In preferred embodiments, the compositions of the present invention are stable, and do not decompose even after long storage periods e. Certain compositions of the present invention are non-toxic and safe when swallowed, inhaled, or contacted to the skin of a host.

This is in contrast to many chemical microbicides, which are known irritants. Additionally, in some embodiments, the compositions are also non-toxic to plants. The compositions of the present invention can be produced in large quantities and are stable for many months at a broad range of temperatures.

Undiluted, they tend to have the texture of a semi-solid cream and can be applied topically by hand or mixed with water. Diluted, they tend to have a consistency and appearance similar to skim milk, and can be sprayed to decontaminate surfaces or potentially interact with aerosolized spores before inhalation.

These properties provide a flexibility that is useful for a broad range of antimicrobial applications. Additionally, these properties make the compositions of the present invention particularly well suited to decontamination applications.

As stated above, at least a portion of the emulsion may be in the form of lipid structures including, but not limited to, unilamellar, multilamellar, and paucliamellar lipid vesicles, micelles, and lamely phases. Some embodiments of the present invention employ an oil phase containing ethanol.

This formulation is highly efficacious against microbes and is also non-irritating and non-toxic to mammalian users and can thus be contacted with mucosal membranes.

In some other embodiments, the emulsions of the present invention comprise a first emulsion emulsified within a second emulsion, wherein a the first emulsion comprises i an aqueous phase; and ii an oil phase comprising an oil and an organic solvent; and iii a surfactant; and b the second emulsion comprises i an aqueous phase; and ii an oil phase comprising an oil and a cationic containing compound; and iii a surfactant.

The following description provides a number of exemplary emulsions including formulations for compositions BCTP and X 8 W 60 PC. X 8 W 60 PC comprises a mixture of equal volumes of BCTP with W 80 8P.

W 80 8P is a liposome-like compound made of glycerol monostearate, refined oya sterols e. The GENEROL family are a group of a polyethoxylated soya sterols Henkel Corporation, Ambler, Pa.

Emulsion formulations are given in Table 1 for certain embodiments of the present invention. These particular formulations may be found in U.

Certain other emulsion formulations are presented in FIG. Moreover, FIG. The X 8 W 60 PC emulsion is manufactured by first making the W 80 8P emulsion and BCTP emulsions separately. A mixture of these two emulsions is then re-emulsified to produce a fresh emulsion composition termed X 8 W 60 PC.

These compounds have broad-spectrum antimicrobial activity, and are able to inactivate vegetative bacteria through membrane disruption. TRITON X 8 vol. Soybean oil NN The compositions listed above are only exemplary and those of skill in the art will be able to alter the amounts of the components to arrive at a nanoemulsion composition suitable for the purposes of the present invention.

Those skilled in the art will understand that the ratio of oil phase to water as well as the individual oil carrier, surfactant CPC and organic phosphate buffer, components of each composition may vary.

Although certain compositions comprising BCTP have a water to oil ratio of , it is understood that the BCTP may be formulated to have more or less of a water phase. For example, in some embodiments, there is 3, 4, 5, 6, 7, 8, 9, 10, or more parts of the water phase to each part of the oil phase.

The same holds true for the W 80 8P formulation. Similarly, the ratio of Tri N-butyl phosphate:TRITON Xsoybean oil also may be varied.

Although Table 1 lists specific amounts of glycerol monooleate, polysorbate 60, GENEROL , cetylpyridinium chloride, and carrier oil for W 80 8P, these are merely exemplary.

An emulsion that has the properties of W 80 8P may be formulated that has different concentrations of each of these components or indeed different components that will fulfill the same function. For example, the emulsion may have between about 80 to about g of glycerol monooleate in the initial oil phase.

In other embodiments, the emulsion may have between about 15 to about 30 g polysorbate 60 in the initial oil phase. In yet another embodiment the composition may comprise between about 20 to about 30 g of a GENEROL sterol, in the initial oil phase.

The nanoemulsions structure of the certain embodiments of the emulsions of the present invention may play a role in their biocidal activity as well as contributing to the non-toxicity of these emulsions.

Adding the oil phase to the detergent and solvent markedly reduces the toxicity of these agents in tissue culture at the same concentrations. While not being bound to any theory an understanding of the mechanism is not necessary to practice the present invention, and the present invention is not limited to any particular mechanism , it is suggested that the nanoemulsion enhances the interaction of its components with the pathogens thereby facilitating the inactivation of the pathogen and reducing the toxicity of the individual components.

It should be noted that when all the components of BCTP are combined in one composition but are not in a nanoemulsion structure, the mixture is not as effective as an antimicrobial as when the components are in a nanoemulsion structure.

Numerous additional embodiments presented in classes of formulations with like compositions are presented below. The effect of a number of these compositions as antipathogenic materials is provided in FIG.

The following compositions recite various ratios and mixtures of active components. One skilled in the art will appreciate that the below recited formulation are exemplary and that additional formulations comprising similar percent ranges of the recited components are within the scope of the present invention.

In certain embodiments of the present invention, the inventive formulation comprise from about 3 to 8 vol. Some of these embodiments comprise PBS. For example, one embodiment of the present invention comprises about 3 vol.

Another similar embodiment comprises about 3. Yet another embodiment comprises about 3 vol. Still another embodiment comprises about 3 vol. A further embodiment comprises about 8 vol. In some embodiments of the present invention, the inventive formulations comprise about 8 vol.

In the present invention, some embodiments comprise from about 8 vol. In certain embodiments of the present invention, the inventive formulation comprise from about 1 to 2 vol. Some of these formulations comprise PBS. It is contemplated that the addition of PBS in some of these embodiments, allows the user to advantageously control the pH of the formulations.

For example, one embodiment of the present invention comprises about 2 vol. In another embodiment the formulation comprises about 1.

In alternative embodiments of the present invention, the formulations comprise from about 5 vol. In still other embodiments of the present invention, the formulations comprise from about 5 vol. In still other embodiments of the present invention, the formulations comprise from about 2 to 8 vol.

For example, the present invention contemplates formulations comprising about 2 vol. In other similar embodiments, the formulations comprise about 3 vol. In still further embodiments, the formulations comprise about 4 vol.

In yet other embodiments, the formulations comprise about 5 vol. Another embodiment of the present invention comprises about 6 vol. In still further embodiments of the present invention, the formulations comprise about 8 vol. In yet another embodiment comprises 8 vol.

In alternative embodiments of the present invention, the formulations comprise from about 1 to 2 vol. Additionally, certain of these formulations may comprise from about 1 to 5 vol. In some of the embodiments comprising liquid baby formula, the formula comprises a casein hydrolysate e.

In some of these embodiments, the inventive formulations further comprise from about 0. Other similar embodiments comprising these basic components employ phosphate buffered saline PBS as the aqueous phase. For example, one embodiment comprises about 2 vol. In still other embodiments, the inventive formulation comprises about 2 vol.

In another similar embodiment, the formulations comprise about 1. In yet another embodiment of the present invention, the formulations comprise about 1.

In still another embodiment, the formulations comprise about 1. In another embodiment of the present invention, the formulations comprise about 1. In some embodiments of the present invention, the inventive formulations comprise about 3 vol.

In a particular embodiment of the present invention, the inventive formulations comprise about 3 vol. In some embodiments of the present invention, the inventive formulations comprise from about 4 to 8 vol.

Additionally, certain of these embodiments further comprise about 1 vol. For example, in certain of these embodiments, the inventive formulations comprise about 8 vol. In another embodiment of the present invention, the inventive formulations comprise about 8 vol.

In still another embodiment, the formulations comprise about 8 vol. In yet another embodiment, the formulations comprise about 8 vol. Another embodiment of the present invention comprises about 4 vol. Still another related embodiment comprises about 8 vol. In other embodiments, the inventive formulations comprise about 8 vol.

In an alternative embodiment of the present invention, the formulation comprise about 8 vol. In another exemplary embodiment of the present invention, the formulations comprise about 8 vol. In still further embodiments, the present invention comprises about 8 vol.

Additional similar embodiments comprise 8 vol. In another embodiment of the present invention, the inventive formulations further comprise about 5 vol. In some embodiments of the present invention, the inventive formulations comprise about 2 vol.

In an additional embodiment of the present invention, the formulations comprise about 8 vol. Certain related embodiments further comprise about 1 vol. For example, one particular embodiment comprises about 8 vol.

In still another embodiment, the inventive formulations comprise about 8 vol. In still further embodiments, the inventive formulations comprise about 8 vol. For example, in one particular embodiment the formulations comprise about 8 vol. Another related embodiment comprises about 8 vol.

In yet other embodiments, the inventive formulations comprise from about 8 vol. In still other embodiments, the present invention comprises about 8 vol.

In another embodiment of the present invention, the formulations comprise about 0. In another embodiment of the present invention, the inventive formulations comprise about 2 vol.

In another related embodiment, the inventive formulations comprise about 2 vol. In still other embodiments of the present invention, the inventive formulations comprise about 8 to 10 vol. Additionally, in certain of these embodiments, the compositions further comprise about 1 vol. An additional related embodiment comprises about 10 vol.

In still another embodiment of the present invention, the inventive formulations comprise about 10 vol. In some embodiments of the present invention, the inventive formulations comprise about 8 to 10 vol.

Exemplary embodiments of such formulations comprise about 8 vol. A related formulation comprises about 8 vol. In yet another embodiment of the present invention, the inventive formulations comprise about 4 vol.

In some embodiments of the present invention, the inventive formulations comprise about 0. In yet another embodiment of the present invention, the inventive formulations comprise about 8 vol.

The specific formulations described above are simply examples to illustrate the variety of compositions that find use in the present invention. The present invention contemplates that many variations of the above formulation, as well as additional nanoemulsions, find use in the methods of the present invention.

To determine if a candidate emulsion is suitable for use with the present invention, three criteria are analyzed. Using the methods and standards described herein, candidate emulsions can be easily tested to determine if they are suitable.

First, the desired ingredients are prepare using the methods described herein, to determine if an emulsion can be formed. If an emulsion cannot be formed, the candidate is rejected. For example, a candidate composition made of 4.

Second, the candidate emulsion should form a stable emulsion. An emulsion is stable if it remains in emulsion form for a sufficient period to allow its intended use. For example, for emulsions that are to be stored, shipped, etc. Typical emulsions that are relatively unstable, will lose their form within a day.

The following candidate emulsions were shown to be stable using the methods described herein: 0. Third, the candidate emulsion should have efficacy for its intended use. For example, an anti-bacterial emulsion should kill or disable bacteria to a detectable level.

As shown herein, certain emulsions of the present invention have efficacy against specific microorganisms, but not against others. Using the methods described herein, one is capable of determining the suitability of a particular candidate emulsion against the desired microorganism.

Generally, this involves exposing the microorganism to the emulsion for one or more time periods in a side-by-side experiment with the appropriate control samples e. In preferred embodiments of the present invention, the nanoemulsions are non-toxic e.

In some embodiments the non-toxic nanoemulsions comprise surfactant lipid preparations SLPs for use as broad-spectrum antimicrobial agents that are effective against bacteria and their spores, enveloped viruses, and fungi. In preferred embodiments, these SLPs comprises a mixture of oils, detergents, solvents, and cationic halogen-containing compounds in addition to several ions that enhance their biocidal activities.

Ingredients for use in the non-toxic nanoemulsions include, but are not limited to:. detergents e. Emulsions are prepared, for example, by mixing in a high shear mixer for minutes.

The emulsions may or may not be heated before mixing at 82° C. for 1 hour. In general, the non-toxic nanoemulsions are characterized by the following: they are nm in diameter; the charge depends on the ingredients; they are stable for relatively long periods of time e.

The specific compositions of the present invention possess a range of beneficial activities and properties. A number of the exemplary beneficial properties and activities are set forth below: A Microbicidal and Microbistatic Activity; B Sporicidial and Sporistatic Activity: C Viricidal and Viralstatic Activity; D Fungicidal and Fungistatic Activity; and E In vivo Effects.

Additionally, FIG. The methods of the present invention can be used to rapidly inactivate bacteria. In certain embodiments, the compositions are particularly effective at inactivating Gram positive bacteria. In preferred embodiments, the inactivation of bacteria occurs after about five to ten minutes.

Thus, bacteria may be contacted with an emulsion according to the present invention and will be inactivated in a rapid and efficient manner.

It is expected that the period of time between the contacting and inactivation may be as little as minutes or less where the bacteria is directly exposed to the emulsion.

However, it is understood that when the emulsions of the present invention are employed in a therapeutic context and applied systemically, the inactivation may occur over a longer period of time including, but not limited to, 5, 10, 15, 20, 25, 30, 60 minutes post application.

Further, in additional embodiments it may be that the inactivation may take two, three, four, five or six hours to occur. In other embodiments, the compositions and methods of the invention can also rapidly inactivate certain Gram negative bacteria.

In some embodiments, the bacteria inactivating emulsions are premixed with a compound that increases the interaction of the emulsion by the cell wall.

The use of these enhancers in the compositions of the present invention is discussed herein below. It should be noted that certain emulsions especially those comprising enhancers are effective against certain Gram positive and negative bacteria and may be administered orally where they will come in contact with necessary gut bacteria.

In specific embodiments, the present invention has shown that the emulsions of the present invention have potent, selective biocidal activity with minimal toxicity against vegetative bacteria. BCTP was highly effective against B. cereus, B. circulans and B. megaterium, C.

perfringens, H. influenzae, N. gonorrhoeae, S. agalactiae, S. pneumonia, S. pyogenes and V. cholerae classical and Eltor FIG. This inactivation starts immediately on contact and is complete within 15 to 30 minutes for most of the susceptible microorganisms.

In certain specific embodiments, the present invention has demonstrated that the emulsions of the present invention have sporicidal activity. Without being bound to any theory an understanding of the mechanism is not necessary to practice the present invention, and the present invention is not limited to any particular mechanism , it is proposed the that the sporicidal ability of these emulsions occurs through initiation of germination without complete reversion to the vegetative form leaving the spore susceptible to disruption by the emulsions.

The initiation of germination could be mediated by the action of the emulsion or its components. The results of electron microscopy studies show disruption of the spore coat and cortex with disintegration of the core contents following BCTP treatment.

Sporicidal activity appears to be mediated by both the TRITON X and tri-n-butyl phosphate components since nanoemulsions lacking either component are inactive in vivo. The present invention demonstrates that mixing BCTP with B.

cereus spores before injecting into mice prevented the pathological effect of B. Further, the present invention shows that BCTP treatment of simulated wounds contaminated with B.

oereus spores markedly reduced the risk of infection and mortality in mice. The control animals, that were injected with BCTP alone diluted , did not show any inflammatory effects proving that BCTP does not have cutaneous toxicity in mice.

These results suggest that immediate treatment of spores prior to or following exposure can effectively reduce the severity of tissue damage of the experimental cutaneous infection. Other experiments conducted during the development of the present invention compared the effects of BCTP and other emulsions derived from BCTP to inactivate different Bacillus spores.

anthracis spores in four hours, and was also sporicidal against three other Bacillus species through the apparent disruption of spore coat. X 8 W 60 PC diluted had more sporicidal activity against B. anthracis, B. cereus, and B. subtilis and had an onset of action in less than 30 minutes.

In mice, mixing BCTP with B. Mortality was reduced 4-fold in the latter experiment. The present compositions are stable, easily dispersed, non-irritant and nontoxic compared to the other available sporicidal agents.

The bacteria-inactivating oil-in-water emulsions used in the methods of the present invention can be used to inactivate a variety of bacteria and bacterial spores upon contact.

For example, the presently disclosed emulsions can be used to inactivate Bacillus including B. megatetium, also including Clostridium e.

botulinum and C. The methods of the present invention may be particularly useful in inactivating certain biological warfare agents e. In addition, the formulations of the present invention also find use in combating C.

perftingens, H. BCTP contains TRITON X while SS and W 80 8P contain TWEEN 60, and NN contained nonoxynol-9 surfactant. Each is a non-ionic surfactant, but differs in its chemistry and biological characteristics.

Nonoxynol-9 has strong spermicidal activity and it is widely used as a component of vaginally delivered contraceptive products Lee, It has been claimed to have virucidal effect against enveloped viruses Hermonat et al.

However, nanoxynol-9 has not been shown to be effective against nonenveloped viruses Hermonat et al. globigii spores. In additional embodiments, it was demonstrated that the nanoemulsion compositions of the present invention have anti-viral properties.

The effect of these emulsions on viral agents was monitored using plaque reduction assay PRA , cellular enzyme-linked immunosorbent assay ELISA , P-galactosidase assay, and electron microscopy EM and the cellular toxicity of lipid preparations was assessed using a 4,5-dimethylthiazoleyl -2,5 diphenyltetrazolium MTT staining assay Mosmann There was a marked reduction of influenza A infectivity of MDCK cells as measured by cellular ELISA with subsequent confirmation by PRA.

BCTP was the most potent preparation and showed undiminished virucidal effect even at dilution Kinetic studies showed that 5 min incubation of virus with BCTP at dilution completely abolished its infectivity. TRITON X, an active compound of BCTP, at dilution only partially inhibited the infectivity of virus as compared to BCTP, indicating that the nanoemulsion itself contributes to the anti-viral efficacy.

To further examine the anti-viral properties of BCTP, its action on non-enveloped viruses was investigated. The BCTP treatment did not affect the replication of lacZ adenovirus construct in cells as measured using β-galactosidase assay. When examined with EM, influenza A virus was completely disrupted after incubation with BCTP while adenovirus remained intact.

Adenovirus treated with different dilutions of BCTP showed no reduction in infectivity. The efficacy of certain BCTP based compositions against various viral onslaught and their minimal toxicity to mucous membranes demonstrate their potential as effective disinfectants and agents for prevention of diseases resulting from infection with enveloped viruses.

Yet another property of the nanoemulsions of the present invention is that they possess antifungal activity. Common agents of fungal infections include various species of the genii Candida and Aspergillus, and types thereof, as well as others.

While external fungus infections can be relatively minor, systemic fungal infections can give rise to serious medical consequences. There is an increasing incidence of fungal infections in humans, attributable in part to an increasing number of patients having impaired immune systems. Fungal disease, particularly when systemic, can be life threatening to patients having an impaired immune system.

Candida was grown at 37° C. Cells were then washed and counted using a hemacytometer. A known amount of cells were mixed with different concentrations of BCTP and incubated for 24 hours. The Candida was then grown on dextrose agar, incubated overnight, and the colonies were counted.

One of skill in the art will be able to take the formulations of the present invention and place them into appropriate formulations for the treatment of fungal disease. The nanoemulsions of the present invention find use in combatting infections such as athletes foot, candidosis and other acute or systemic fungal infections.

Animal studies demonstrated the protective and therapeutic effect of the present compositions and methods. Bacillus cereus infection in experimental animals has been used previously as a model system for the study of anthrax See e.

The disease syndrome induced in animals experimentally infected with B. cereus is similar to anthrax Drobniewski, Clin. Experiments conducted during the development of the present invention demonstrated that mixing BCTP with B.

Further, it was demonstrated that BCTP treatment of simulated wounds contaminated with B. cereus spores markedly reduced the risk of infection and mortality in mice. The control animals, which were injected with BCTP alone diluted , did not show any inflammatory effects proving that BCTP does not have cutaneous toxicity in mice.

In a particular example, Guinea Pigs were employed as experimental animals for the study of C. perftingens infection. perftingens without any further treatment. Another group was infected with the same number of bacteria, then 1 hour later it was irrigated with either saline or BCTP to simulate post-exposure decontamination.

Irrigation of experimentally infected wounds with saline did not result in any apparent benefit. However, BCTP irrigation of the wound infected with C. perfingens showed marked reduction of edema, inflammatory reaction and necrosis.

As such, it was demonstrated that certain formulations of the present invention can be used to combat a bacterial infection. All rats in the oral toxicity study showed weight gain over the study period.

No adverse clinical signs were noted and all tissues appeared within normal limits on gross examination. Bacterial cultures from the stools of treated animals were not significantly different from those of untreated animals.

Set forth below are a number of exemplary uses for the compositions disclosed herein: A Pharmaceuticals and Therapeutics; B Decontamination and Sterilization; C Food Preparation; and D Kits, as well as a description of methods and systems for the E Modification, Preparation, and Delivery of the compositions of the present invention.

Such compositions may be employed to reduce infection, kill microbes, inhibit microbial growth or otherwise abrogate the deleterious effects of microbial infection. For in vivo applications, the compositions can be administered in any effective pharmaceutically acceptable form to warm blooded animals, including human and animal subjects.

Generally, this entails preparing compositions that are essentially free of pyrogens, as well as other impurities that could be harmful to humans or animals. Particular examples of pharmaceutically acceptable forms include but are not limited to oral, nasal, buccal, rectal, vaginal, topical or nasal spray or in any other form effective to deliver active compositions of the present invention to a site of microorganism infection.

In preferred embodiments, the route of administration is designed to obtain direct contact of the compositions with the infecting microorganisms. In other embodiments, administration may be by orthotopic, intradermal, subcutaneous, intramuscular or intraperitoneal injection.

The compositions may also be administered to subjects parenterally or intraperitonealy. Such compositions would normally be administered as pharmaceutically acceptable compositions. Except insofar as any conventional pharmaceutically acceptable media or agent is incompatible with the emulsions of the present invention, the use of known pharmaceutically acceptable media and agents in these particular embodiments is contemplated.

In additional embodiments, supplementary active ingredients also can be incorporated into the compositions. For topical applications, the pharmaceutically acceptable carrier may take the form of a liquid, cream, foam, lotion, or gel, and may additionally comprise organic solvents, emulsifiers, gelling agents, moisturizers, stabilizers, surfactants, wetting agents, preservatives, time release agents, and minor amounts of humectants, sequestering agents, dyes, perfumes, and other components commonly employed in pharmaceutical compositions for topical administration.

Tablet and dosage forms of the compositions in which the emulsions are formulated for oral or topical administration include liquid capsules, and suppositories.

In solid dosage forms for oral administration, the compositions may be admixed with one or more substantially inert diluent e.

In another embodiment of the invention, the compositions of the invention may be specifically designed for in vitro applications, such as disinfecting or sterilization of medical instruments and devices, contact lenses and the like, particularly when the devices or lenses are intended to be used in contact with a patient or wearer.

For example, the compositions may be used to cleanse and decontaminate medical and surgical instruments and supplies prior to contacting a subject. Additionally, the compositions may be used to post-operatively, or after any invasive procedure, to help minimize the occurrence of post operative infections.

In especially preferred embodiments, the compositions are administered to subjects with compromised or ineffective immunological defenses e. For applications of this type, the compositions may be conveniently provided in the form of a liquid, foam, paste or gel and may be provided with emulsifiers, surfactants, buffering agents, wetting agents, preservatives, metal ions, antibiotics and other components commonly found in compositions of this type.

In other embodiments, the compositions may be impregnated into absorptive materials, such as sutures, bandages, and gauze, or coated onto the surface of solid phase materials, such as surgical staples, zippers and catheters to deliver the compositions to a site for the prevention of microbial infection.

Other delivery systems of this type will be readily apparent to those skilled in the art. The compositions can also be used to treat other internal and external microbial infections e.

simplex, etc. In these applications, the emulsions can be formulated with therapeutic carriers as described above. In certain embodiments, the antimicrobial compositions and methods of the present invention also include a variety of combination therapies.

For example, often single antimicrobial agents are much less effective at inhibiting microbes than are several agents employed in conjunction with each other. This approach is often advantageous in avoiding the problems encountered as a result of multidrug resistance. This is particularly prevalent in bacteria that have drug transporters that mediate the efflux of drugs from the organism.

The present invention further contemplates the use of the present methods and compositions in such combination therapies. There are an enormous amount of antimicrobial agents currently available for use in treating bacterial, fungal and viral infections. Hardman et al. McGraw Hill, chapters 43 through 50, , herein incorporated by reference in its entirety.

Generally, these agents include agents that inhibit cell wall synthesis e. Various combinations of antimicrobials may be employed. Actual amounts of compositions and any enhancing agents in the compositions may be varied so as to obtain amounts of emulsion and enhancing agents at the site of treatment that are effective in killing vegetative as well as sporular microorganisms and neutralizing their toxic products.

Accordingly, the selected amounts will depend on the nature and site for treatment, the desired response, the desired duration of biocidal action and other factors. Generally, the emulsion compositions of the invention will comprise at least 0.

It is envisioned that viral infections may be treated using between about 0. Bacterial infections may be attacked with compositions comprising between about 0. Spores can be killed by emulsions comprising from about 0.

These are merely exemplary ranges. It is envisioned that the formulations may comprise about 0. It should be understood that a range between any two figures listed above is specifically contemplated to be encompassed within the metes and bounds of the present invention.

Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.

Moreover, for human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by the FDA Office of Biologics standards. In general, the present invention contemplates compositions and methods that find use as environmental decontamination agents and for treatment of casualties in both military and terrorist attack.

The inactivation of a broad range of pathogens, including vegetative bacteria and enveloped viruses See e. Preferred compositions of the present invention can be rapidly produced in large quantities and are stable for many months at a broad range of temperatures.

These properties provide a flexibility that is useful for a broad range of decontamination applications. For example, certain formulations of the present invention are especially effective at destroying many of the bacterial spores and agents used in biological warfare.

In this regard, the compositions and methods of the present are useful in decontaminating personnel and materials contaminated by biological warfare agents. Solutions of present compositions may be sprayed directly onto contaminated materials or personnel from ground based, or aerial spraying systems.

In certain of these applications, the present invention contemplates that an effective amount of composition be contacted to contaminated materials or personnel such that decontamination occurs. Alternatively, personal decontamination kits can be supplied to military or civilians likely to become contaminated with biological agents.

Disease [] , combined with low toxicity makes the present compositions particularly well suited for use as general decontamination agents before a specific pathogen is identified. Thus, certain embodiments of the present invention specifically contemplate the use of the present compositions in disinfectants and detergents to decontaminate soil, machinery, vehicles and other equipment, and waterways that may have been subject to an undesired pathogen.

Such decontamination procedures may involve simple application of the formulation in the form of a liquid spray or may require a more rigorous regimen. Also, the present emulsions can be used to treat crops for various plant viruses in place of or for use with conventional antibiotics. Nanoemulsions may also be used to decontaminate farm animals, animal pens, surrounding surfaces, and animal carcasess to eliminate, for example, noneveloped virus of hoof and mouth disease.

In addition to their use in decontamination of land and equipment, the formulations also find use in household detergents for general disinfectant purposes. Moreover, some embodiments of the present invention can be used to prevent contamination of food with bacteria or fungi e.

This can be done either in the food preparation process, or by addition to the food as an additive, disinfectant, or preservative. The inventive emulsions are preferably used on hard surfaces in liquid form.

Accordingly, the foregoing components are admixed with one or more aqueous carrier liquids. The choice of aqueous carrier is not critical. However, it should be safe and it should be chemically compatible with the inventive emulsions.

In some embodiments, the aqueous carrier liquid comprises solvents commonly used in hard surface cleaning compositions. Such solvents should be compatible with the inventive emulsions and should be chemically stable at the pH of the emulsions.

Solvents for use in hard surface cleaners are described, for example, in U. In preferred embodiments, the aqueous carrier is water or a miscible mixture of alcohol and water.

The alcohol can be used to adjust the viscosity of the compositions. In some embodiments, the alcohols are preferably C2-C4 alcohols. In particularly preferred embodiments, ethanol is employed. The present invention also embodies non-liquid compositions.

These non-liquid compositions can be in granular, powder or gel forms, preferably in granular forms. Optionally, some compositions contain auxiliary materials that augment cleaning and aesthetics so long as they do not interfere with the activity of the inventive emulsions.

The compositions can optionally comprise a non-interfering auxiliary surfactant. A wide variety of organic, water-soluble surfactants can optionally be employed.

The choice of auxiliary surfactant depends on the desires of the user with regard to the intended purpose of the compositions and the commercial availability of the surfactant.

Detergent builders can also be employed in the compositions. Detergent builders sequester calcium and magnesium hardness ions that might otherwise bind with and render less effective the auxiliary surfactants or co-surfactants.

Detergent builders are especially useful when auxiliary surfactants or co-surfactants are employed, and are even more useful when the compositions are diluted prior to use with exceptionally hard tap water e.

In other embodiments, the composition further comprise, suds suppressors. In these embodiments, the compositions preferably comprise a sufficient amount of a suds suppressor to prevent excessive sudsing when contacting the compositions to hard surfaces.

Suds suppressors are especially useful in formulations for no-rinse application of the composition. The suds suppressor can be provided by known and conventional means. Selection of the suds suppressor depends on its ability to formulate in the compositions, and the residue and cleaning profile of the compositions.

The suds suppressor must be chemically compatible with the components in the compositions, it must be functional at the pH range described herein, and it should not leave a visible residue on cleaned surfaces. Low-foaming co-surfactants can be used as suds suppressor to mediate the suds profile in the compositions.

Examples of suitable co-surfactants for use herein include block copolymers e. The optional suds suppressor preferably comprises a silicone-based material. These materials are effective as suds suppressors at very low concentrations.

At low concentrations, the silicone-based suds suppressor is less likely to interfere with the cleaning performance of the compositions. An example of suitable silicone-based suds suppressors for use in the compositions is Dow Corning DSE.

These optional but preferred silicone-based suds suppressors can be incorporated into the composition by known and conventional means. In still other embodiments, the compositions may be used by health care workers, or any persons contacting persons or areas with microbial infections, for their personal health-safety and decontamination needs.

In addition, the inventive emulsions can be formulated into sprays for hospital and household uses such as cleaning and disinfecting medical devices and patient rooms, household appliances, kitchen and bath surfaces, etc. In similar embodiments, the compositions may be used by sanitation and environmental services workers, food processing and agricultural workers and laboratory personnel when these individuals are likely to contact infectious biological agents.

Additionally, the compositions may be used by travelers and persons contacting ares likely to harbor infectious and pathological agents. The present invention also contemplates that certain compositions described herein may be employed in the food processing and preparation industries in preventing and treating food contaminated with food born bacteria, fungi and toxins.

Thus, such compositions may be employed to reduce or inhibit microbial growth or otherwise abrogate the deleterious effects of microbial contamination of food.

For these applications, the emulsion compositions are applied in food industry acceptable forms such as additives, preservatives or seasonings. Except insofar as any conventional additives, preservatives and seasonings are incompatible with the emulsions of the present invention, their use in preventing or treating food born microbes and their toxic products is contemplated.

Supplementary active ingredients may also be incorporated into the compositions. For such applications, acceptable carriers may take the form of liquids, creams, foams, gels and may additionally comprise solvents, emulsifiers, gelling agents, moisturizers, stabilizers, wetting agents, preservatives, sequestering agents, dyes, perfumes and other components commonly employed in food processing industry.

In another embodiment of the present invention, the compositions may be specifically designed for applications such as disinfecting or sterilization food industry devices, equipment, and areas where food is processed, packaged and stored.

For applications of this type, the compositions may be conveniently provided in the form of a liquid or foam, and may be provided with emulsifiers, surfactants, buffering agents, wetting agents, preservatives, and other components commonly found in compositions of this type.

In some embodiments, the compositions are applied to produce or agricultural products prior to or during transportation of those goods.

Compositions of the invention may be impregnated into absorptive materials commonly used in packaging material for the prevention of food contamination during transport and storage e. Actual amounts of the emulsions and enhancing agents in the compositions of the invention may be varied so as to obtain appropriate concentrations of emulsion and enhancing agents to effectively prevent or inhibit food contamination caused by food born microbes and their toxic products.

Accordingly, the selected concentrations will depend on the nature of the food product, packaging, storage procedure and other factors. In particular embodiments, emulsions can be used as disinfectants and detergents to decontaminate and prevent microbial infection of food, soil and water, machinery and other equipment, and animals.

The inventive emulsions can be used by the food industry to prevent contamination. For example, inclusion of the emulsion within the food product itself would be effective in killing bacteria that may have been accidentally contaminated meat or poultry.

This could also allow the industry to use a potentially broader spectrum of food products and reduce costs. Certain embodiments of the present invention can also be used in the beverage industry. For example, the inventive emulsions could be included in juice products to prevent growth of certain fungi, which cause contamination and lead to production of mycotoxins, which are dangerous to consumers.

Through the addition of small amounts of the inventive emulsions, the most common fungal contaminants in fruit juice were prevented.

Lionel A. The adverse Herbal tea blends reactions associated with Herbal tea blends have become a Non-toxiv of major importance and High blood sugar symptoms in the Non-toxic antimicrobial agents Non-tlxic years. Non-toxic antimicrobial agents toxicity Muscle building for beginners take many forms, varying gaents mild, transient phenomena to dramatic, life-threatening events such as seizures or cardiac arrhythmias. Agebts review the toxicity of antimicrobials in general and of the fluoroquinolones in particular and attempt to explain the adverse events by use of structure-adverse event relationships where possible. There are currently 5 main mechanisms that can be invoked to explain antimicrobial toxicity: direct effects, hypersensitivity, changes in microbial flora, drug interactions, and microbial lysis. The adverse drug reactions seen with fluoroquinolones are explained on the basis of these 5 mechanisms. The various organ systems affected by the fluoroquinolones are considered; then individual members of the fluoroquinolone class are reviewed.