An In Vitro Investigation Of The Antifungal Potency Of 7% Zinc Pyrithione Versus 2% Ketoconazole Against Malassezia

Background:Dandruff is a chronic scalp problem that needs constant attention to alleviate the scaling and itchy symptoms.It is a relapsing dermatologic condition that is caused by a fungus called"Malassezia"which colonizes the scalp,it can be treated by using medicated shampoo.Humans and many domestic and wild animals have Malassezia yeasts as part of their typical cutaneous microbiota.They invade skin by entering the sebaceous glands as they’re lipophilic fungi.^[1]They are opportunistic microorganisms that can cause a variety of skin illnesses.Pityrosporum,folliculitis is one of the most well-known Malassezia fungus forms.When yeast molecules get into the hair follicles and grow,they cause skin infection that looks like acne.^[1,2]Teenagers and young adults are the most typically affected by it.Fortunately,the infection can be treated with topical oral,or shampoo antifungal medicines,which contains antifungal ingredients such as ketoconazole（KET）,zinc pyrithione（ZPT）,terbinafine,nystatin…etc.Although there are many antifungal agents that helps treat the infection,KET and ZPT are the two of the most well-known antifungal options which comes in shampoo form.^[2]Both function by stopping the fungus from growing by limiting the creation of ergosterol,the fungal analogue of cholesterol,which increases membrane fluidity and prevents the fungus from growing.Although both medications are efficient at suppressing fungal activity and growth,one of them is thought to be superior in terms of effectiveness and time to notice results.^[3,4]Objective:The aimed of this research was to investigate the in vitro antifungal efficacy of 7%ZPT compared with 2%KET against Malassezia.The study employed the hole-punch method and microdilution method to investigate the antifungal efficacy of the two drugsUsing those methods,the minimum inhibitory concentration（MIC）of the drugs was determined,which is the lowest concentration of a drug that can inhibit the growth of a microorganism.It is important to consider that the choice of solvent and dilution factor may affect the drug’s solubility,stability,and activity.Since Malassezia strain was used for this research,DMSO（dimethyl sulfoxide）was used to dilute the antifungal agents.Method:The experiment was conducted by placing the Malassezia strain Malassezia furfur（previously known as P.ovale）fungus on Sabouraud agar culture plate and was intubated at 37℃ for 48 hours.The sample were transferred to Sabouraud broth and re-incubate them for another 24 hours.The stock solution was prepared by dissolving 10 mg of the 2%KET and 20 mg of7%ZPT in 1 m L of DMSO giving a stock solution of 10 mg/m L KET and 20mg/m L ZPT was obtained.Stock solution was promptly dispersed in tubes and was further diluted with distilled water.The concentrations ranging from 0.001 to 100μg/ml.A 7 mm diameter hole created on each SDA plate,and the fungal strain was spread on the surface of the plate using Sterile swabs,including the hole.A volume of 100μL of the antifungal solution was introduced into the well and incubated at a temperature of 32℃ for a period of 14 days.On day 4,day 7,and day 14,the diameter of the inhibition zone was measured.Results:Results indicated that ZPT had better results when it came to%fungal activities.The average antifungal activity of ZPT was found to be 32.1%at0.001μg/ml,27.4%at 0.01μg/ml,25.0%at 0.1μg/ml,23.1%at 1.0μg/ml,and 18.1%at 10μg/ml.At 100μg/ml,the antifungal activity was 0%for all contact times.Similarly,the average antifungal activity of KET was found to be 25.9%at 0.001μg/ml,25.7%at 0.01μg/ml,24.5%at 0.1μg/ml,16.7%at1.0μg/ml,and 8.4%at 10μg/ml.At 100μg/ml,the antifungal activity was0%for all contact times.The comparison of inhibition zones between ZPT and KET was a critical aspect of the treatment of the fungal strain.The objective of the experiment was to compare the antifungal effectiveness of ZPT and KET by measuring their inhibition zone diameters against a culture of fungi.To achieve this,various concentrations of each antifungal agent were prepared and added to the fungal culture.Following a 14-day exposure to the antifungal agents,the diameter of the inhibition zone was determined,and the mean and standard deviation for each concentration were computed.The findings revealed that both ZPT and KET had higher antifungal activity at higher concentrations and longer exposure periods,as demonstrated by larger inhibition zones.Overall,ZPT had higher antifungal activity than KET.Statistical analysis was performed to compare the efficacy of these two drugs,using a one-tailed test.Based on Table 3 p-value obtained was around0.002,and the two-tailed p-value was at 0.003.The one-tailed p-value was preferred because of the prior expectation that one drug would perform better than the other.With a p-value less than 0.05,the results indicate a statistically significant difference between the inhibition zones of ZPT and KET,confirming that ZPT is more effective than KET in inhibiting the growth of the fungus.These findings are important for clinicians when deciding on the most effective treatment options for patients with Malassezia-related skin conditions.Conclusion:In conclusion,the results showed that the difference between the KET group and the ZPT group was statistically significant.It should be emphasized that this study was performed in vitro,and additional investigations are required to establish the efficacy of KET and ZPT in vivo.The results of this study provide valuable insights into the antifungal efficacy of two commonly used medications,ZPT and KET against Malassezia.While these treatments hold promise,their long-term effectiveness and safety remain uncertain,additional research to fully explore their potential benefits and drawbacks is unquestionable.New and more effective antifungal agents need to be identified to treat dandruff and other skin infections caused by Malassezia.Future studies should also investigate the molecular mechanisms underlying the antifungal activity of these medications,which could lead to the development of more targeted and personalized therapies for individuals with chronic scalp problems.Acquiring a deeper understanding of the pathogenesis of Malassezia and other fungal pathogens could be a game-changer in the realm of fungal infections.It has the potential to revolutionize the way these infections are diagnosed,prevented,and treated in both humans and animals.With this knowledge,new and innovative approaches could be unlocked providing multiple ways to combat fungal infections.