Comparative Studies on Antibacterial Ef icacy of Natural & Arti icial Honey on Isolated Pathogens from Burnt Wound Sample

Vijaya Benerji G*1, Anantha Lakshmi Y2, Shankar K3, Punitha V C3 1Department of Biochemistry, Konaseema Institute of Medical Sciences Research Foundation, Amalapuram, Andhra Pradesh, India 2Department of Surgery, Konaseema Institute of Medical Sciences Research Foundation, Amalapuram, Andhra Pradesh, India 3Department of Community Medicine, Meenakshi Academy of Higher Education and Research, Chennai, Tamil Nadu, India


INTRODUCTION
Honey may be made toward bees Likewise a nourishment wellspring. Done cool climate or At nourishment sources need aid scarce, bees use their put away honey Likewise their sourball for vitality. Toward contriving to bees swarms should home in simulated hives, individuals have been ready with semi-domesticated those insects Also harvest abundance honey. In the hive, the bees use their "HONEY STOMACH" on ingest And spew the honey a few times until it is incompletely digested (Bergman et al., 1983). Bees store honeycombs and there utilize it for sustenance for winter. Since it is a regular sweetener on account of it will be high glucose Furthermore fructose content; it will be generally utilized within candies, cereals, furthermore heated merchandise.
There are more than 300 sorts from claiming honey's luctuating on lavor and shades (from pale yellow with dim amber), relying upon the sort of blossoms visited toward those honey bumblebee. Honey need been referred to on people for hundreds of years to bring an antimicrobial potential (Amor, 1978).
The vast majority of microorganisms don't develop done honey due to its low water movement of 0. 6% (Aneja, 2017). That improvement for wound contamination is not another wonder and is a progressing issue to numerous individuals. Open wounds serve concerning illustration an entrance course to sullying 28 for attacking microorganisms.
Furthermore, deferred wound recuperating of contaminated wounds; it may be connected with expanded discomfort, doctor's facility stay, also medicinal services costs (Bulman, 1955). Contaminated wounds scar that's only the tip of the iceberg extremely also would connect with prolonged rebuilding. That hazard of systemic contamination furthermore actually passing may be additionally connected with contaminated. Wounds (Cooper et al., 2002). (Cooper et al., 1999) The Clinical sample (Burnt Wound) collected aseptically was from The Government Hospital from Kumbakonam, Tanjore Dt (Tamilnadu). The Sterile swab might have been acquainted profoundly suf icient will get An damp example, and the swab might have been swapped in the transport networking. The sampled swab might have been made promptly of the lab for preparing of the example. Whether deferred inoculated in the supplement stock.

Preparation about networking furthermore plating method
Seclusion about clinical tests might have been streaked again the supplement agar plates and incubated at 30 o C to 48 hours. After those brooding period, chose states of clinical tests were exchanged from those blended society of the plate onto particular agar plates and incubated toward 30 o C to 48 hours. Slants holding immaculate society were put away In 4 o C for further examination.

Morphologic and physiological test
Social aspects from claiming immaculate detaches previously, supplement agar networking were recorded after brooding time to 300c from claiming 48 hrs. Morphologic systems comprise of perceptible also minute routines. The minute characterization might have been carried out by light microscopy. Eventually, Tom's perusing utilizing that minute technique manifestation from claiming creatures might have been distinguished similarly as color, state and morphological tenet (Efem, 1993).

Gram Staining
Gram staining strategy might have been Initially ran across by a danish doctor named Hans Christian gram clinched alongside 1884. This strategy isolates bacterial phones under two major groups, gram certain Furthermore, gram-negative bacteria, this makes it a crucial device to elucidation Also separation for microorganisms (Fabry et al., 1998).

Accumulation about honey example
Honey tests were gathered starting with two diverse districts.., Natural (paruthicheri), and Arti icial (shop), in sterile containers and kept at room temperature (24-26 0 c) for 30 days before experimentation. Each sample was diluted to various concentrations (25%, 50%, 75% and 100% in v/v) in order to test their antibacterial potential.

Preparation of honey at dilution natural honey & arti icial honey
The preparation of Natural & Arti icial honey in various concentration (100%, 75%, 50%, 25%) In 100% Natural & Arti icial honey consist of pure honey without dilution. 75% Natural & Arti icial honey consists of 7.5 ml of Natural & Arti icial honey diluted with 2.5 ml of distilled water. 50% Natural & Arti icial honey consists of 5 ml of honey diluted with 5 ml of distilled water. 25% Natural & Arti icial honey consists of 2.5 ml of honey diluted with 7.5 ml of distilled water.

Determination of antibacterial effect of honey sample: preparation of disc
Whatman No.1, 6mm iltered paper disc was organized and pasteurized by autoclaving (Ward and Safle, 1995). These discs are coated, and each disc is saturated by 15 ml diffusion concentration of honey and dried overnight at 31 o C.

Natural honey & arti icial honey
Muller Hinton agar plates were ready, and the surface of the plate might have been permitted will dry. Every Muller-Hinton agar plate might have been inoculated for those standard inoculums similar to e. Coli, staphylococcus aureus, Pseudomonas aeruginosa by splashing An swab Furthermore pivoting it In those agar plate. That regular nectar under the ixation from claiming 25%, 50%, 75% Furthermore, 100% might have been dipped under that circle. The characteristic nectar circle might have been set In the inoculated agar. Following 24 hours for brooding toward 37oC, the Zone from claiming restraint might have been measured Also recorded. A triplicate plate might have been additionally conveyed out.

Anti-microbial affectability test 11 circles dispersion system
The chemotherapeutic operators would know as antibiotics. Every anti-microbial do have distinctive antimicrobial exercises concerning diverse microorganisms. A portion of them bring a short-range A percentage for wide range against extensive variety from claiming microbes.

Pathogenic bacteria
The pathogenic bacteria were isolated from the Burnt wound infection by using the swab technique and isolated in nutrient agar medium (Figure 1). Then the isolated bacteria were identi ied through a gram staining method (Table 1) and some various biochemical tests (Table 2).

Identi ication of pathogenic bacteria
The isolated organisms were identi ied by using the grams staining method and using the selective medium for the organisms. The result showed two gram-negative organisms and one gram-positive organism.
The organisms were con irmed by using the Eosinmethylene agar medium, Blood agar medium, cetrimide agar medium. In Eosin methylene agar medium on E.coli shows metallic green colonies in the Blood agar medium on Staphylococcus aureus shows pink color colonies. In cetrimide, agar medium on Pseudomonas aeruginosa shows green color colonies.

Effect of arti icial honey on -Escherichia coli
Arti icial honey showed the inhibitory action against Escherichia coli by 25% 50% 75% concentration is 0mm. But in 100% concentration of honey shows 11 mm of inhibition.

Staphylococcus aureus
Arti icial honey showed the inhibitory action against Staphylococcus aureus by 25% 50% 75% concentration is 0mm. But in 100% concentration honey shows 9 mm of inhibition.

Pseudomonas aeruginosa
Natural honey showed the inhibitory action against Pseudomonas aeruginosa by 25%50% 75% concentration is 0mm.
But in 100% concentration honey shows 8 mm of inhibition. Write given in Table 4.

Effect of antibiotics on -Escherichia coli
The antibiotic disc showed the inhibitory action against Escherichia coli by Ampicillin is 8mm, Ceftriaxone is 11mm, Erythromycin is 16mm, Penicillin is 13mm, Tetracyclinis 8mm.
The comparative result of antibiotic discs were illustrated as a bar diagram in Figure 3.
As of late this need been rediscovered What's more nectar will be On reasonably totally spread use Similarly as An topical anesthesia antibacterial agenize to those medicine about wounds, blazes Also skin ulcers, there being huge numbers reports from claiming its adequacy in Figure 4. Those exhibits consider higher action about nectar against staphylococcus aureus, escherichia coli. What's more p. Aeruginosa in Figure 5. The available investigation higher action about nectar against staphylococcus aureus, escherichia coli Furthermore p.Aeruginosa in Table 5. The antibacterial action might have been exhibited for both gram-positive Also gram-negative microscopic organisms. Gram-negative microscopic organisms were that's only the tip of the iceberg powerless on nectar. This agrees for An investigation Toward that nectar need that's only the tip of the iceberg action against both gram-positive over gram-negative living being in Table 6. Those helter-skelter antimicrobial action found backing in this examines against the gram-negative microscopic organisms tried alongside the control strains. This might have been obvious in the rate levels of bacterial affectability concerning illustration secondary concerning illustration 100% to. E.coli. In my present study, the antibacterial activity of honey shows a higher inhibition compared with synthetic antibiotics in Table 7. The synthetic antibiotics like Ampicillin, Ceftriaxone, Erythromycin, Penicillin and Tetracyclin in Table 8.

CONCLUSIONS
The present study dealt with the Isolation and Characterization of a bacterial pathogen from a burnt wound sample. The burnt looped infection is a main dif iculty in patients. Burnt wounds presented meaningfully higher bacterial counts associated by the normal wound. The burnt wound sample was inoculated using nutrient agar medium and medium and incubated at 37 0 C for 24 hrs. After incubation pure cultures of organisms were made in respective agar mediums. The present work on the honey showed antibacterial activity against the burnt wound pathogenic Bacteria such as Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. Natural honey and arti icial honey showed maximum inhibition zone was formed in 100% honey against pathogens. When compared with the zone of inhibition against pathogens for natural and arti icial honey, the natural honey showed higher effective than arti icial honey against the burnt wound pathogens. The tetracycline shows the maximum inhibition against Pseudomonas aeruginosa than other antibiotics. It will meet the requirements of essential products required in humans, animals as well as in the pharmaceutical industry. The natural honey is more effective and safe than artiicial honey and the antibiotics against the burnt wound pathogens. Finally, from my study, I concluded that natural honey generally good medicine for burn wound infections.