INTRODUCTION: The term biocide is increasingly being used to describe compounds with antiseptic, disinfectant or, sometimes, preservative

activity. A compound might be used in only one such capacity or possess two or even all of these properties. Until fairly recently, there were two longheld general opinions about biocides. The first was that, as long as they were effective, there was little reason (apart from academic value) to determine how they achieved their inhibitory or lethal effects. The second, widely perceived, view was that antiseptics and disinfectants acted as general protoplasmic poisons and, as such, merited little attention. At the beginning of the twentieth century, there were few drugs available for the treatment of infections. Antiseptics and disinfectants had at that stage been employed for various purposes and in various guises, notable examples being phenol (carbolic acid), mercuric chloride, chlorine, hypochlorites and iodine. Quaternary ammonium compounds (QACs) were described in 1916 but were not used commercially for another 19 years or so. Early studies on the action of such compounds concentrated on the kinetics of bacterial inactivation, although Cooper notably described the relationship between phenolics (phenol and meta-cresol) and bacterial proteins as being of importance in their mechanism of disinfection. In particular, it was considered that the protein structure inside the bacterial cell was affected. Subsequently, Knaysi et al. ,reported on the manner of death of bacteria, mainly Escherichia coli, exposed to mild chemical and physical agents, concluding that the order of death was determined by the distribution of resistances among the cells. Later, Jordan & Jacobs found the distribution of resistance of E. coli treated with phenol to be normal at all phenol concentrations used. Interestingly, specific enzymes were considered by some workers,, to be involved in bacterial inactivation by biocidal agents. Penicillin (benzylpenicillin, penicillin G) began to be used as a chemotherapeutic antibiotic in the 1940s. Early studies on its mode of action were undertaken by Gardner and Duguid, and Eagle & Musselman demonstrated a paradoxical effect of high concentrations on staphylococci. However, it was not until the isolation of the Park nucleotides and subsequent studies, especially those by Strominger et al.,,, that it was realized that not only was penicillin an important antibiotic but also that it was a valuable tool in studying bacterial peptidoglycan biosynthesis. Further discoveries, the production of other -lactam antibiotics and work

demonstrating that tetracyclines and other antibiotics inhibited bacterial protein synthesis, provided a stimulus for additional and comprehensive investigations to be undertaken on bacterial inactivation mechanisms (and also, naturally, on bacterial resistance mechanisms) by such selectively toxic drugs. It was realized that these aspects, linked to structureactivity relationships, provided the key to the development of many new, improved antibiotic molecules. Until recently, there has not been the same enthusiasm for studying the mode of action of biocidal agents. However, the literature does contain a surprising, if scattered, number of publications about the mechanisms of inhibition and inactivation of Gram-positive nonsporulating bacteria (although the information on mycobacteria is disappointing), bacterial spores (as well as germinating and outgrowing ones) and Gram-negative bacteria, Less data are available about the mechanisms of fungal and viral inactivation, with very few comprehensive studies on the mechanisms of protozoal inactivation.,It is rather surprising that information on the mechanisms of viral inactivation, in particular, is sparse. Furthermore, it is not known why the MICs of biocides such as chlorhexidine and QACs are of an equivalent order for both mycobacteria and staphylococci whereas these cationic biocides possess low mycobactericidal potency but are rapidly lethal to the latter. Contrary to early suggestions by Rahn et al., it is now considered to be highly unlikely that bacterial cells possess a single type of target enzyme the inhibition or inactivation of which by a biocide is responsible for a loss of cell viability. Thus, although triclosan has repeatedly been shown to inhibit enoyl reductase (see below), work from this laboratory has demonstrated that other events are involved in bacterial inactivation. It has become clear that some antiseptics and disinfectants on the one hand and antibiotics on the other have similar effects on bacteria. For example, (1) filament formation is induced in Gramnegative bacteria by both antibiotics (-lactams, novobiocin, fluoroquinolones) and biocides (phenoxyethanol, phenylethyl alcohol, chloroacetamide, acridines); (2) inhibition of enoyl reductase, involved in fatty acid synthesis, is inhibited by both isoniazid, an important antitubercular drug, and the bisphenol (phenylether) triclosan; and (3) autolysis brought about by low concentrations of phenolics and inorganic and organic mercury compounds has been suggested as being similar to that following bacterial exposure to penicillin. However, it is interesting that E. coli cells exposed to phenoxyethanol, proflavine or chloroacetamide showed the same susceptibility to ampicillin and norfloxacin as cells not pre-exposed to a biocide. It

would be interesting to determine whether biocide-resistant E. coli cells showed differences in response to these important antibiotics. A correlation was found earlier by McKellar et al. as a consequence of a non-specific increase in impermeability. Additionally, it must be noted that the end result may be brought about by different inhibitory or lethal mechanisms. It has also become increasingly obvious that insusceptibility mechanisms to biocides and antibiotics may be similar although not necessarily identical. A natural (intrinsic) insusceptibility to both groups may be shown by Gram-negative bacteria and mycobacteria, with outer membrane or cell wall impermeability being responsible. Additionally, efflux systems may remove toxic drug and biocide molecules from the cell, although a key issue here relates to the concentrations at which the compounds are used. Thus, with drugs it is usually necessary to equate MICs or minimum bactericidal concentrations (MBCs) with blood serum levels. By contrast, biocides are essentially used for external purposes at concentrations likely to be considerably higher than MICs or MBCs; such concentrations are unlikely to be effluxed from bacterial cells. Nevertheless, the possibility remains that low, residual concentrations could act as a focus for the survival of organisms containing efflux genes or for the gradual or rapid development of biocide-resistant bacteria. This reinforces the argument that the effects of biocides on bacterial (and, indeed, other types of microbial) cells should be examined over a wide range of concentrations. There are other reasons for studying the mechanisms of biocides. At present, comparatively little is known about the uptake of biocides into bacterial (and other microbial) cells. The probability exists that targeted drug delivery, whereby a biocide can readily reach its target site(s) within a cell, could lead to greater efficacy. An aspect that needs to be considered is the possible design of new biocidal molecules based on known effects of current molecules; there is little evidence that this is happening. The only significant new biocides to be introduced in the past few years have been ortho-phthalaldehyde (OPA) and those based on peracetic acid. However, both OPA and peracetic acid are themselves old molecules that have been examined in a new, antimicrobial context. In recent years, rotation of disinfectants in hospitals and elsewhere, e.g. in the pharmaceutical and food industries, has been advocated to prevent the development of bacterial resistance. It has been claimed that, ideally, one disinfectant should be replaced by another having a dissimilar

mechanism of action.Clearly, a knowledge of the ways in which such agents act is an essential component of such a policy. In conclusion, there is an urgent need to investigate more fully the nature of the inhibitory and lethal effects of antiseptics and disinfectants on a range of microorganisms and microbial entities. Possible multiple target sites and concentration-dependent effects would form an important aspect of such studies, which would also provide a better understanding of intrinsic and acquired bacterial resistance mechanisms and of the possible linkage between biocide usage and antibiotic resistance.

OBJECTIVE:

To evaluate the effectiveness of antiseptic and disinfectant. To identify the best antiseptics and disinfetants can be used to inhibit the growth of staphylcoccus aereus and esterechia coli. MATERIALS: Broth containing staphylococcus aereus Broth containing esterechia coli Chlorohexidine 4% Clorox (HOCL) Antibacterial soap Povidone Potasium permanganate (KMNO4) Nutrient agar Filter paper Puncture Foreceps Paraffin Cotton swab Parafilm

METHOD: 33 students were divided into five groups and each group were given two agar plates. PROCEDURE: 1. The nutrient agar were labelled accordingly A1 and A2. 2. The nutrient agar plates were streaked with staphylococcus aereus using sterile cotton swab. 3. Then, the filter paper was punched and rinse with chlorhexidine 4%, then the paper was placed at the centre of the streaked nutrient agar using foreceps. 4. Later, the nutrient agar plate was sealed with parafilm. 5. Step 2 to 4 was followed to the broth containing esterichia coli. 6. The rest of the antiseptic and disinfectant materials were continued by another group with similar procedure. 7. All the nutrient agar plates were placed in incubator for 24hours at 37c. 8. The results were observed and recorded.

RESULTS: Types Of Antiseptic And Disinfectant. Chlorhexidine 4% Clorox(Hocl) Potassium Permanganate(KmNo4) Hydrogen Peroxide(H2O2) Povidone 9mm 20mm No Growth 29mm Zone Of Inhibition(mm). Staphylococcus Aereus 14mm 38.1mm No Growth Esterechia Coli 32mm 42mm No Growth

PICTURES:

Povidone

Staphylococcus Aereus.

E.Coli

Chlorhexidine 4%

Staphylococcus Aereus.

Chlorox(HoCl)

E.Coli

Staphylococcus Aereus.

E.Coli

Hydrogen Peroxide(H2O2)

E.Coli .

Staphylococcus Aereus.

E.Coli. DISCUSSION:

Staphylococcus Aereus.

In Staphylococcus Aereus the growth of Zone of Inhibition decreases from Clorox(Hocl) with 38.1mm, then Povidone 20mm,Chlorhexidine 4% 14mm, Hydrogen Peroxide(H2O2) and there is no growth of Zone of Inhibition in Potassium Permanganate(KmNo4).So this shows that the Potassium Permanganate(KmNo4) is the best antiseptics can be used for Staphylococcus Aereus, whereas Clorox is less effective disinfectants compare to other agent tested and has large Growth of Zone of Inhibition. In the other hand, the Growth of Zone of inhibition decreases from Clorox(Hocl) with 42mm, then Chlorhexidine 4% 32mm, Povidone 29mm and no Growth of Zone of Inhibition in both Hydrogen Peroxide(H2O2) and Potassium Permanganate( KmNo4 ) in E.Coli. As the result, Hydrogen Peroxide(H2O2) and Potassium Permanganate( KmNo4) is the best antiseptics can be used for E.Coli , whereas Clorox(HoCl) is less effective disinfectants compare to other agent tested and has large Growth of Zone of Inhibition. When compare to the all the antiseptics and disinfectants tested, Potassium Permanganate (KmNo4) is the best antiseptic can be used against both Staphylococcus Aereus and in E.Coli, although in E.Coli there is no Growth when tested with Hydrogen Peroxide(H2O2) but it is less effective against Staphylococcus Aereus and there is Growth of Zone of Inhibition. In Contrast, (HoCl) is less effective disinfectants can be used against Staphylococcus Aereus and E.Coli because there is large Growth of Zone of Inhibition in both agars. CONCLUSION: As the conclusion (KmNo4) is the best antiseptic can be used against both Staphylococcus Aereus and in E.Coli whereas, (HoCl) is less effective disinfectants can be used against Staphylococcus Aereus and E.Coli.