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Assessment of Antimicrobial Activity of Cerumen (Earwax) and Antibiotics

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Assessment of Antimicrobial Activity of Cerumen (Earwax) and Antibiotics Microbiology Research 2020; volume 11:8565 Assessment of antimicrobial nation of fatty acid). Another gland i.e. apoc- activity of cerumen (earwax) rine sweat glands release secretion that com- Correspondence: Iffat Naz, Department of bines with the sebum to form cerumen. It Biology, Scientific Unit, Deanship of and antibiotics against picks up discarded cells, ear follicles and Educational Services, Qassim University, pathogenic bacteria isolated may contain dust or other debris, but the Buraidah, 51452, Qassim, Kingdom of Saudi from ear pus samples resulting compound forms earwax or ceru- Arabia (KSA). men.6 Tel.: +966 533897891 E-mail: [email protected] ; There are two different types of geneti- [email protected] Iffat Naz cally determined earwax such as wet type Department of Biology, Scientific Unit, and dry type. The dry type is most common Key words: Ear pus samples; Pathogenic bac- Deanship of Educational Services, in Asians and Native Americans and has a teria; Antibiotics; Cerumen; Antibacterial Qassim University, Buraidah, Qassim, grey brownish colour while the wet type can potency. Kingdom of Saudi Arabia be found in Caucasians and Africians and has a brown or dark colour.7 About 30-50% of Acknowledgements: The author would like to South Asians, Central Asians and Pacific thanks Mr. Jamil & Miss. Javeria (M. Phil islanders have the dry type of cerumen. Scholars), for assisting in collection of ear pus Abstract Cerumen type has been used by anthropolo- samples from different hospitals of Peshawar, gist to track human migratory patterns, such KP, Pakistan. She is also grateful to Dr. Abdul The present study is focused on the Rehman for providing laboratory facilities, at as those of Eskimas.8 Further, the study of assessment of the antimicrobial activity of Department of Microbiology, Abasyn cerumen and antibiotics against bacteria earwax has shown controversy, as some University Peshawar, KP, Pakistan. Moreover, isolated from ear pus samples. Thus, a total authors have suggested that earwax has high the author has deep appreciation for Dr. Saleh of 50 ear pus samples were collected from level of nutrients which help in microbial S Alhewairini (Dean) Deanship of Educational infected patients using sterile swabs and growth and other hold a view that earwax has services, Qassim University, KSA for giving were screened using pure culture tech- some compounds that inhibit the growth of her an opportunity to conduct her research niques. Total of 04 different bacterial iso- microorganisms and prevent ear infec- onlyactivities smoothly. 9,10 lates were identified while, the prevalence tions. Ear infection in any form has a var- Conflict of interest: The Author declares no data revealed that Pseudomonas spp., were ied microbial etiology, which influences the potential conflict of interests. dominant (58%, n = 29) among isolated selection of an efficacious of anti-microbial agents. According to WHO survey, 42 mil- bacteria followed by Staphylococcus spp., Received for publication: 2 April 2020. lion people worldwide have hearinguse loss (22%, n = 11), Escherichia coli (14%, n = 7) Accepted for publication: 19 April 2020. and Proteus spp., (6%, n = 3). Further, where major cause is otitis media. Infection bioassay revealed that Pseudomonas spp., of the ear can be classified depending upon This work is licensed under a Creative and Staphylococcus spp., were most sensi- the site: otitis externa (infection of external Commons Attribution NonCommercial 4.0 tive to Clindamycin (94.73%) while dis- ear) and otitis media (infection on middle License (CC BY-NC 4.0). ear).11 Otitis is a bacterial infection of the ear played resistant to Ciprofloxacin and ©Copyright: the Author(s), 2020 Ampicillin. Similarly, E. coli and Proteus canal caused by rupture in the normal skin or cerumen, which is a protective barrier in the Licensee PAGEPress, Italy spp., were most sensitive to Ciprofloxacin Microbiology Research 2020; 11:8565 presence of elevated humidity and tempera- (92.8-95.21%) as compared to the other 12 doi:10.4081/mr.2020.8565 antibiotics. Moreover, antibacterial activity ture. It is commonly known as swimmer’s of cerumen was also assessed against test ear, though anything that disrupts this protec- tive lipid layer can lead to the introduction fied the Microbacterium species organisms and its maximum activity was 13 observed against Pseudomonas spp., (90% and proliferation of bacteria. Trauma from (Microbacterium otitidis and equivalent to Clindamycin potency) and cleaning the ears with fingernails or cotton Microbacterium alconae) as key pathogens buds has been identified as the most common 17 Staphylococcus spp., (60% equivalent to in acute otitis externa. The present research predisposing factor locally.14 Moreover, the Amoxicillin potency) while least effective study was designed to isolate and identify two most common bacterial species isolated against E. coli (36%) and ProteusNon-commercial spp., use pathogeniconly bacteria from the infected ear pus from the external auditory canal of normal (22%). Thus, it was concluded that the anti- clinical samples and then, to estimate the individuals are the Staphylococcus species bacterial activity of cerumen might be due antimicrobial activity of cerumen and antibi- (Staphylococcus auricularis, Staphylococcus to the presence of potential chemicals i.e. otics against the isolated pathogenic bacteria. epidermidis, Staphylococcus aureus and flavonoids and terpenoids. Staphylococcus capitis) and the Corynebacterium species (Turicella otitidis Materials and Methods and Corynebacterium auris). The third most Introduction frequently recovered bacteria are the Sample collection Earwax (cerumen) is a grey brown or Streptococci and Enterococci. Together, they A total of 50 ear pus samples were collect- yellowish waxy substance secreted in the account for more than 90% of the normal ed from infected patients admitted in tertiary outer one third of ear canal and, is a protec- flora in the external auditory canal.14,15 care hospital at district Peshawar, KPK, tive barrier between the external environment Pseudomonas aeruginosa, S. epidermidis Pakistan, using sterile swabs under proper and deep external auditory canal.1 Chemical and S. aureus are the most common patho- hygienic conditions. The samples were labeled composition of earwax includes 60% desqua- genic bacteria isolated in acute diffuse otitis bearing the person code, age, sex and date. mated, 12-20% saturated and unsaturated externa locally.16 Doree and Burton,17 stud- Collected samples were suitably diluted and fatty acid and 6-10% cholesterol.2-5 It is a ied 2039 subjects with acute otitis externa were streaked on nutrient media. After incuba- secretion of specialized sets of glands, like and isolated similar bacterial species. tion, these were subjected for isolation of path- sebaceous glands that secrete sebum (combi- Further, the same research group also identi- ogenic bacteria using standard technique. [Microbiology Research 2020; 11:8565] [page 5] Article Cerumen collection Flavonoid determination sion method. Nutrient agar plates were pre- Earwax or cerumen samples were col- About 2-3 ml of aqueous extract of ceru- pared and approximately, 50 µl inoculum of lected from both male and female healthy men was taken in labelled test tubes and then every selected pathogenic bacterial specimen persons of approximately same age groups, 2% NaOH was added to it drop by drop. were homogeneously spreaded on specified using sterile ear buds and the amount of ceru- After this, concentrated HCl was added to the plates with the help of glass spreader. After men was measured in milligram (mg). Then solution. If color of the solution was disap- the period of five minutes, three wells of the samples were pooled and dissolved in pearing after addition of HCl, then it indicat- about 8 mm diameter were bored in the 18 sodium bicarbonate-glycerol buffer (pH 8.2) ed the presence of flavonoids. media plates, one well was used for positive at 1-5% concentration. control (antibiotic), one well was used for Phenolic compounds determination negative control (distilled water) while the Isolation and identification Aqueous extract of cerumen was used for remaining one was used for earwax extract. determination of phenolic compounds The equal volume (50 µL) of controls as well of pathogenic bacteria through Ferric chloride test. Almost 1 ml of as extract were poured into these wells. The Isolation and identification of pathogenic aqueous extract was mixed with 2 ml dis- plates were incubated for 24 h at 37°C. After bacteria from the collected samples were car- tilled water and only a few drops of 10% incubation, zones of inhibition were meas- ried out using standard bacteriological proce- Ferric chloride was added to it. If blue and ured in millimeter (mm) to evaluate the dures i.e. cultural characteristics, microscopy green color appeared after addition of Ferric potency of earwax extract against test patho- and different biochemical tests (catalase, oxi- chloride, then it is indication of the presence genic bacterial specimens. dase, urease, indole, coagulase and triple of phenolic compounds and if there was no sugar iron tests) according to the Berges th coloration then the test was considered as Manual of Determinative Bacteriology 9 negative for phenolic compounds.18 Results Edition. Terpenoids determination Bacteriological analysis of ear pus Antibiotic susceptibility assay Aqueous extract of cerumen was used for samples and prevalence of isolated
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