The Effect of Manuka Honey on Elastase Production by P Aeruginosa - Research Paper Example

?The effect of Manuka honey on elastase production by P. aeruginosa Discussion Manuka honey is widely used in treatment of wounds. This is a tradition that has been practiced since ancient times. Wide studies have been conducted on the inhibitory effect of manuka honey on P. Aeruginosa. Manuka honey Leptospermum polygaliform is a New Zeeland product of manuka bush. The honey is widely preferred since its cheap (Adebolu, 2005). Apart from its effective response in treatment of wounds the honey is also widely accepted by scientists since it has no side effects on wounds. Pseudomonas species tend to be very resistant to most of antimicrobial therapies. During P. aeruginosa infections, P. aeruginosa elastase is released. This is a zinc metalloproteinase released during the process of infection. Any dead wound that is perfused allows multiplication of bacteria to multiply and elaborate an array of virulence. This factor can slow the process of healing in a wound. Protection of P. aeruginosa in the wound occurs through host immune effectors. This can grow at very sufficient levels to elaborate toxins that break down host factors. P. aureginosa has shown potential inactivation of human plasma alpha 1- proteinase. Favorable conditions of 25 degrees in temperature at a molar ratio of E/I= 1: 100 shows potential deactivation within an hour (El-Sukhon, 1994). The factors in target at this point are the fibrin and collagen molecules which have the mandate of healing issues and regeneration leading to complete healing of wounds. For this, purpose many scientist view pseudomonas as a delaying factor in the process of healing in the wounds. These syntheses are required for the synthesis of the auto inducer and the molecules which are linked with it like lactones and also homoserine. The pathogenesis linked with chronic infection along with the importance linked with quorum sensing remains unknown. P.aeruginosa is basically isolated from the patients of CF and the patients producing auto inducers and also the production is linked with CF sputum which is linked directly with the aeruginosa biofilms. The auto inducers are present in lung tissue of the infected mice especially with the P.aeruginosa. Many scientists have experienced honey used to heal wounds. The main function of honey is the stimulation of cytokine production by the monocytes present in the body of the organism. The cytokine production is accelerated hence leading to tissue repair and healing of wounds. The viscosity of manuka honey provides a moist environment, which helps in providing the protective barrier. In addition to this, honey has mild acidity and low levels of hydrogen peroxide stimulate the healing process. Studies related to this indicate that strong solutions of honey and sugar continue to inhibit microbial growth due to the presence of high osmolarity potentials (Lusby, 2005). The presence of high levels of hydrogen peroxide in manuka honey provides a good base for healing of wound and it activity in strains of P. aeruginosa. The experimental results show manuka honey having the potential production of broad-spectrum antibacterial activity. This process takes place by honey removing malodor. The anti-inflammatory activity of the manuka honey helps in reducing edema and consequently exudation and minimizing scarring. This process has the potential of stimulating the growth of granules. The granulation in addition to growth of epithelial tissues shows significant improvement in aiding the process of healing. The experiment shows that manuka honey has minimum inhibitory concentration of around 6% against P. aeruginosa strains. Further research by Nzaeko and Hamdi (Nzaeko BC, 2000) indicate that strains of E. coli and P. aeruginosa show inhibition levels of up to 40%. However, our study shows a significantly lesser percentage but the theory and conclusion is relatively common. P. aeruginosa produces an enzyme elastase, which has been shown to be one of the main virulent factors of these bacteria. This enzyme is one of the main enzymes to cause the destruction of elastin fibers in tissues hence their destruction as observed in many infections caused by this organism e.g. corneal destruction in eye infection and destruction of lung tissue in pulmonary infection. Elastase requires an optimum pH of 7.5, which is slightly alkaline for its activity and cause tissue destruction. Lowering the pH of wounds to slightly acidic or lower than what is required by the enzyme to work could be an option to avoid infections caused by this organism. Manuka honey, when used in dressings lowers the pH of the wound thus inhibiting elastase activity to some extent. Pseudomonas aeruginosa is basically a major bacterial pathogen which is basically existent in patients which are suffering from cystic fibrosis or also known as diffuse panbronchiolitis. Azithromycin which is basically a macrolides are normally not included in basically antipseudomonal therapeutic and also arsenal which is basically due to the absence of bacteriostatic or the bactericidal activity (Molan, 2004). When talking about bacterial organisms, their virulence is very important for them in order to invade the host and cause disease. Many bacteria produce different substance in order to increase their virulence. Some of which are protective capsules to prevent antibiotic drugs enter the bacterial cell and enzymes such as beta lactamase to degenerate the beta lactam ring in penicillin, collagenase and protease etc. P. aeruginosa produces an enzyme elastase which has been shown to be one of the main virulent factors of these bacteria. This enzyme is one of the main enzymes to cause the destruction of elastin fibers in tissues hence their destruction as observed in many infections caused by this organism e.g. corneal destruction in eye infection and destruction of lung tissue in pulmonary infection (Moudoi, 2005).The following table shows the results obtained after adding manuka honey at different concentrations on P. aeruginosa. Strain Honey Added % Abs 1st 2nd 3rd Average Abs 8626 0 0.132 0.128 0.129 0.129 8626 7.5 0.173 0.152 0.149 0.151 8626 15 0.174 0.176 0.176 0.176 867 0 0.108 0.095 0.096 0.096 867 5 0.108 0.095 0.096 0.096 867 10 0.150 0.191 0.189 0.190 Form the table we can see the strain 8626 at 0% averagely yields 0.129 Abs. The inhibition level increases significantly in the strain with addition of 7.5%, which yields 0.151, Abs. doubling the concentration at 15% does not double the average inhibition but raises it significantly to 0.176. We can therefore conclude that increase in concentration of manuka honey leads to further inhibition and delay in microbial activity. The factors behind the inhibition are hydrogen peroxide, flavonoids, and phenolic acids and many others inhibitory factors found in manuka honey (Scmidtchen, 2003). Scientists regard the post antibiotic effect and time-to-kill as very important information since they act as pharmacodynamics parameters in clinical areas thereby providing information. This information is necessary since its useful in determining the therapeutic regime in response to the increasing prevalence of bacterial species which are resistant to antibiotics. Scientist need this information in order to e=determine the level of doses to be adminitsred due to resistance of any atibioic. Increasing dosage may lead to the bcterias developing resisnatance against the treatment. This information is necessary while conducting research on the effect of manuka honey on P. aeruginosa. Hydrogen peroxide leads to major post antibiotics effects. Removal of this component from manuka honey will definitely ensure that effect on the organism is reduced or removed completely. Concentrations of hydrogen peroxide, osmaolarity, and non-reoxide differs in different types of honey. Research indicates that neither does non-peroxide nor that of hydrogen peroxide antibacterial activities of manuka honey increases with concentration of the honey. Results indicate a slight change between changes in concentration from 5% to 20%. Despite much speculation and research on effects of honey on microbial activities, it is still not evident on the mechanisms and processes that manuka honey induces and acts on post antibiotics effects of the antibacterial agents such as P. aeruginosa. Scientist blame this fact on the difference in morphology and the process of metabolism which differs in many microorganisms. For example, an observation from an electron microscope shows increasing numbers of cross walls and cell thickness in a class of antibiotics mainly P. aeruginosa and staphylococci. This difference has been show to delay the microbial activity in E. coli by 4 hours when the two organisms are placed under the similar antimicrobial conditions. Overall wall thickness in P. aeruginosa can also be increased in the presence of para-minobenzoic acid. It is seen that 2 ug of azithromycin basically inhibits the quorum sensing circuitry which is linked with the pseudomonas aeruginosa strain which is called PAO1. Synthetic auto inducers in addition to it are partially restored which then helps in expressing the transcriptional activator which is involved in encoding. Gram positive and gram-negative bacterial growth is also inhibited in the presence manuka honey. Strain 867 showed similar results as that of 8626. Increase in concentration of the honey averagely increases the inhibitory levels of microbial activities. There is a reduction of the auto inducers and the production which is linked with 50 ug of erythromycin which has been specially suggested. The chromobacterium which is used helps in measuring the auto inducers. The major concern is whether there is an interference of azithromycin along with the quorum sensing circuitry or not. The differentiation takes places through the inhibition linked with the virulence factors linked with the production from non specific effects which can be used in optimal way, in the conditions which are experimental with the stationary phase being on set, especially keeping in view the quorum sensing which stays active and is ready for growth This result is consistent with other studies that show inhibitory factors of manuka honey on P. aeruginosa due to the thermal stability of the glucose oxidase enzyme, peroxide factors, plant and floral factors used in production of the honey in addition to increased levels of hydrogen peroxide in manuka honey (Wilkinson, 2005). Despite wider contributions of hydrogen peroxide, its use in the process of healing wounds due to its nature of causing inflammation in wounds. There is a reduction of the auto inducers and the production which is linked with 50 ug of erythromycin which has been specially suggested. The chromobacterium which is used helps in measuring the auto inducers. The major concern is whether there is an interference of azithromycin along with the quorum sensing circuitry or not. The differentiation takes places through the inhibition linked with the virulence factors linked with the production from non specific effects which can be used in optimal way, in the conditions which are experimental with the stationary phase being on set, especially keeping in view the quorum sensing which stays active and is ready for growth. In treatment of cystic fibrosis, there is need for transcription activator LasR so that elastase I fully expressed. Further research indicates that gene lasl is important for the process of high expression of elastase. Synthesis of diffusible molecules of Pseudomonas auto inducer (PAI) actively involves the presence of lasl gene. It has been shown that PAI provides P. aeruginosa with a means of cell-to-cell communication, which is necessary for the expression of virulence genes. In addition to this, there is also the provisional target for the therapeutic approaches for these genes (Venkatachalam, 2007). The moment tissues are deprived of oxygen through obstruction of circulation, xanthine is produced. This enzyme helps in alteration of dehydrogenase to catalyze the oxygen. It is important to understand and measure the effect of azithromycin which has various codes linked with the structural protein and the belongings of the secretion pathway which is being used. Over all the results are very helpful in suggesting that the azithromycin can be very beneficial in reducing the production linked with the quorum sensing dependent factors which are used for inhibiting the synthesis of the molecules which are linked with auto inducers. Debriding action of manuka honey helps in providing a clean granulating wound bed. From the experiment, we can determine several causes of errors that can be used as precautions for after research. Using the same concentration of the honey is very important while conducting the experiment since different concentration levels produces different effects. It is also important to ensure that a sterile environment is cultivated during the process of experimentation. This is to ensure that no contamination occurs thereby leading to reliable results. The biological factors such as cell phase, inoculum intensity, and changes in the process of bacterial morphology could also be a source of error hence this needs to be checked while conducting this experiment in order to arrive at accurate results. Machinery used during the process of conductingg this research is also a significant factor that can cause errors especially on the growth detection and incubating temperatures. Cell mass is directly related to the optical density and therefore a confusion may arise when comparing the optical density with the number of cells. We can therefore advice that comparison of post antibiotic effect obtained from the experiments should be standardized. Conclusion Honey is a very important and powerful substance, it is basically a topical antimicrobial agent which is used in order to cure millions of wounds and it is used for more than millennia in wound care. In 1999 the wound care products which contained medical grade honey especially the licensed product became available and now it is widely and strongly used (Willix, 1999). The properties of therapeutic honey are referring towards them going through the anti inflammatory along with antimicrobial activities. The review which is linked thus provides a special insight linked with the laboratory proof and the publishing which is carried out in the previous five years and how the mechanism basically works of the impact of honey on the wounds and these are then understood by everyone in a better way. Bibliography Adebolu, T. (2005). Effect of natural honey on local isolates of diarrhea causing bacteria in southwestern Nigeria. African Journal of Biotechnology, 4(10):1172-1174. El-Sukhon, S. N.-H. (1994). Effects of honey on Bacterial Growth and Spore Germination. J. Food Prot, 57(10): 918- 920. Lusby, P. A. (2005). Bactericidal activity of different honeys against pathogenic bacteria. Arch. Med. Res, 36:464–46. Molan, P. a. (2004). Clinical usage of honey as a wound dressing: an update. J. Wound Care, 13: 353–6. Moudoi, M. O.-Z. (2005). Antimicrobial activity of honey against food pathogens and food spoilage micro-organisms. Department of Food Science and Technology, Cornell University, NYSAES, vol. 1: 61-71. Nzaeko BC, H. J. (2000). Anti Micobial Potential of Honey on some Microbial Isolates. Journal of Sceintific Research and Medicinal Science, 2: 75-79. Scmidtchen, A. E. (2003). Elastase– producing Pseudomonas aeruginosa degrade plasma proteins and extracellular products of human skin and fibroblasts, and inhibit fibroblast growth. Microb. Pathog, 34: 47–55. Venkatachalam, M. a. (2007). Bactericidal Activity of Different Types of Honey Against Clinical and Environmental Isolates of Pseudomonas aeruginosa. J. Appl. Bacteriol, 13(4):439–441. Wilkinson, J. a. (2005). Antibacterial activity of 13 honeys against Escherichia coli and Pseudomonas aeruginosa. J. Med. Food, 8(1):100-3. Willix, D. P. (1999). A comparison of the sensitivity of wound-infecting species of bacteria to the antibacterial activity of manuka honey and other honeys. J. Appl. Bacteriol, 73: 388-94. Read More