1,4 Department of Applied Microbiology, Faculty of Science, Ebonyi State University, P.M.B. 053, Abakaliki, Ebonyi State.
2,3 Department of Microbiology, Faculty of Biological Sciences, Alex Ekwueme-Federal University Ndufu-Alike Ikwo, P.M.B.1010, Abakaliki, Ebonyi State, Nigeria.
The increasing reports of antimicrobial resistance in healthcare systems, and the need to control microbial growth in hospitals, industries and homes has led to increased production of different types of disinfectants and antiseptics. These antiseptics are often used to sterilize the surfaces of some hospital equipment and other surfaces, thereby reducing their microbial load. This study was therefore aimed at ascertaining the antibacterial activities of commonly used antiseptic liquids in Abakaliki metropolis, against frequently isolated Gram-positive bacterium. A total of 74 samples which includes, 40 skin swabs from male and female medical students and 34 samples from hospital equipment were collected in a tertiary care hospital in Abakaliki, Ebonyi State. The samples were analyzed following standard microbiology procedures. Different concentrations (100, 50, 25 and 12.5 mg/ml) of four antiseptic liquids (Purit, Z germicide, Hypo and Dettol) were prepared following standard procedures. Disk diffusion techniques were used to ascertain the antibacterial activities of the antiseptic liquids against strains of Staphylococcus aureus. Of the 40 skin swabs analyzed, the prevalence of Staphylococcus aureus was 17(85%) and 14(70%) for male and female students respectively. The total prevalence of Staphylococcus aureus among the hospital equipment was 20(58.8%). The study further showed that the activeness of these antiseptics was concentration dependent as highest activity was observed at 100mg/ml and lowest at 12.5mg/ml. Purit and Dettol showed increased activity against the isolated Staphylococcus aureus and the activeness of the antiseptics increases with the increase in their concentration.
The largest organ in the body is the skin and forms the outer coverage of the whole body system, playing great role in keeping the internal tissues free from invading microorganisms and other harmful substances. These great roles are achieved by forming physically protective water proof bi-layer which blocks the entry of these invading pathogens such as parasites, fungi, viruses and bacteria.1 The skin is therefore the first line of defense from invading microorganisms and most bacteria such as Pseudomonas aureginosa and Staphylococcus aureus reside on the skin and become the major cause of various skin infections.2 Every human has different complement of resident or friendly bacteria on their skin surface and over 180 different strains of bacteria reside on the skin surface. These bacteria are mainly members of Staphylococcus Spp. such as S. aureus, S. epidermidis and S. hominis. Others include; Arcanobacterium haemolyticum, Micrococcus luteus, Propionibacterium acnes, Dermabacter group, Corynebacterium group and Brevibacterium spp.3 Often times, transient bacteria such as Pseudomonas aeruginosa, Bacillus subtilis and S. aureus may also be deposited on skin surfaces from environmental sources and are capable of causing various degrees of skin and wound infections. 4,5,6 Several other friendly bacteria species are known to normally reside and cover various areas of human skin and are referred to as the normal flora of the skin. These normal floras of the skin protect the skin by covering all the available spaces thereby preventing other harmful invading bacteria from growing and manifesting on the body of the individual.
Staphylococcus aureus has been reported as the most frequently isolated pathogen within the hospital environment.4,7 This is partly because S. aureus is a normal flora of human body and can easily be transmitted from humans to fomites and vis-visa, thus becoming an important pathogen resulting to various degrees of human infections.7,8
Antiseptics however, plays crucial role in infection control across various settings including but not limited to healthcare facilities, households and industries. These agents are designed to inhibit or destroy microorganisms on living tissues or surfaces, thereby reducing the risk of infections. Antiseptics are used widely in households, industries, healthcare settings and various workplaces with the sole aim of controlling microbial growth either by inhibition or direct killing of invading microorganisms. Antiseptics possess antibacterial activity against vast groups of microorganisms and therefore have the ability to inhibit, destroy or kill the growth of various microorganisms. Antiseptics can possess cidal or static effects depending on its ability to either kill or inhibit microbial growth,4 and this is important in the choice of antiseptic with respects to human body in preventing sepsis and skin infections. Generally, antiseptics hold promise alternatives to other numerous antimicrobial agents targeted at infection control. However, there is paucity of information on comprehensive investigations and efficacy of various antiseptic liquids sold in Abakaliki metropolis and its environs against bacterial strains despite huge patronage and usage enjoyed by these products. Therefore, it becomes imperative to study the effectiveness of some selected antiseptic liquids sold in Abakaliki metropolis as this study will provide an in-depth knowledge of the effectiveness and efficiency of these antiseptics against the most worrisome resident bacteria and pathogenic microorganisms such as Staphylococcus aureus.
MATERIALS AND METHODS
Study area
This study was carried out in National Obstetric Fistula Centre, Abakaliki (NOFIC). This hospital was established by Federal Government of Nigeria in 2011 as the first National health facility for treatment, rehabilitation, research and training of obstetric fistula. The hospital was initially called Mother and Child Care Initiative (MCCI), which was a pet project of her excellency, Chief Mrs Josephine Nwuzor Elechi, the wife of the second executive Governor of Ebonyi State,
Sample Collection
Antiseptic liquids used
The four (4) antiseptic liquids used in this study (Purit, Z germicide, Hypo and Dettol) were purchased from standard pharmaceutical stores within Abakaliki metropolis, Ebonyi State. The batch numbers, expiry date and the presence or absence of the manufacturer’s seal were also noted.9
Hospital equipment sampled
A total of six (6) hospital equipment were sampled. These include; hospital stretchers, surgical table, blood pressure monitor, ventilators, bed rails and thermometers.
Collection of skin swabs
A total of forty (40) skin swabs were collected from male (20) and female (20) medical students of medicine and surgery Department of Ebony State University, Abakaliki. Verbal consent of the students were first sought and obtained before the collection of the samples. The arms of the consented students were swabbed with sterile moisten swab sticks and carefully covered. While a total of 34 samples were collected from hospital equipment: 3 samples from hospital stretchers, 3 sample from surgical table, 5 samples from blood pressure monitor, 3 samples from ventilators, 10 samples from bed rails and 10 samples from thermometers. The surfaces of these hospital equipment were carefully swabbed with moisten swab sticks and were covered immediately. All the 60 samples collected were taken to the laboratory unit of Applied Microbiology Department, Ebonyi State University, for analysis.
Media Preparation
All the media used for this study were prepared following manufacturers’ instructions. The media were mixed properly and sterilized by autoclaving at 121 OC for 15 mins. The sterilized media were allowed to cool to body temperature before about 20 mls were carefully poured on each petridish and allowed to solidify.10
Isolation and identification of S. aureus
The swabs were then inoculated onto already prepared 5 ml nutrient broth and incubated at 37°C for 24 hours. After overnight incubation, the samples were subsequently cultured in mannitol salt agar (MSA) and further incubated overnight at 37oC for 24 hours.10 The bacterial isolates were characterized and identified by colonial appearance/morphology, microscopic examination and several conventional biochemical tests.
Preparation of 0.5 McFarland turbidity standard
The 0.5 McFarland turbidity standard equivalent of the test organism was prepared by constituting one percent raw sulphuric acid in 0.5g of dehydrated barium chloride (BaCl2.2H2O) in 50 ml of distilled water in different reaction flasks respectively. Barium chloride solution (0.6ml) was then added to 99.4ml of the H2SO4 solution in a separate test tube and mixed to obtain 0.5 McFarland turbidity standards.11
Standardization of test inoculum
Each of the isolate was standardized using colony suspension method.12 The test organisms from nutrient agar plates, initially incubated at 37 0C overnight were suspended in nutrient broth and the density of each isolate suspension was continuously adjusted and matched with 0.5 McFarland standards.11
Preparation and dilution of the antiseptic liquids
Using a sterile syringe, 1 ml of each antiseptic was pipetted unto sterile test tube containing 9 ml of sterile water and properly mixed to give 100 mg/ml. From the 100 mg/ml dilution, it was further diluted into different concentration to the 2nd test tube (50 mg/ml), 3rd test tube (25 mg/ml) and 4th test tube (12.5 mg/ml).13
Preparation of paper disks
Six (6) mm was cut from Whattman paper with the aid of 6 mm cork borer and a sterile surgical blade. The disks were then foiled and autoclaved following standard procedures. The disks were then impregnated with different concentrations (100, 50, 25 and 12.5mg/ml) of antiseptic liquids by soaking the paper disks onto different concentrations for about 1 hr using sterile forceps.
Antimicrobial susceptibility testing of the test organism to antiseptic liquids
A 0.5 McFarland turbidity standard equivalent of S. aureus isolates were streaked on the surface of molten Mueller Hinton agar plates using sterile swab sticks. The inoculated plates were allowed for about 20 minutes for pre-diffusion. Antimicrobial susceptibility testing of test organism to antiseptic liquids was tested using disk diffusion methods. Briefly, with sterile forceps, each of the impregnated disk with different concentrations of the antiseptics were aseptically placed 15 mm apart on the surface of well streaked surfaces of Muller Hinton agar. The plates were then incubated overnight at 37oC. After overnight incubation, the visible clear zones of inhibition were measured using a transparent meter ruler according to CLSI guidelines.14
Ethical consideration and sponsorship.
The authors do not in any way tend to use any of the antiseptic liquids for any form of litigation, neither do the authors tend to devalue the antiseptics tested. This study is entirely conceived, sponsored and carried out by the authors, only for advancement of scientific knowledge and deepening scientific study especially in the area of medical microbiology.
RESULTS
Table 1, of the total 40 samples collected from male and female medical students the percentage prevalence of S. aureus among male and female students were 17 (85%) and 14 (70%) respectively. The total prevalence of S. aureus among the medical students was 31(77.5%)
Table 2, among the samples collected from hospital equipment, the percentage prevalence of S. aureus occurrence were 3(100%), 3(100%), 2(40%), 2(66.7%), 6(60%) and 4(40%) for hospital stretchers, surgical tables, blood pressure monitors, ventilators bed rails and thermometers respectively. While the total prevalence of S. aureus among the hospital equipment was 20(58.8%).
Table 3, the antibacterial activities of antiseptics; Purit, Z germicide, Hypo and Dettol against S. aureus isolated from male students showed geometric decrease in the effectiveness of the antiseptics as the concentration decreases. Hence, the inhibition zone diameter (IZD) of Purit, Z germicide, Hypo and Dettol liquids at 100 mg/ml were 21, 17, 22 and 20 mm respectively. While at the lowest concentration of 12.5 mg/ml, the IZD of Purit, Hypo and Dettol liquids decreased to 13, 13 and 12 mm respectively. While the isolate showed resistant to Z germicide. The antibacterial activities of antiseptics; Purit, Z germicide, Hypo and Dettol liquids against S. aureus isolated among female students showed geometric decrease in the effectiveness of the antiseptics as the concentration decreases. Hence, the inhibition zone diameter (IZD) of Purit, Z germicide, Hypo and Dettol liquids at 100 mg/ml was 18, 16, 20 and 18 mm respectively. While at the lowest concentration of 12.5 mg/ml, the IZD decreased thus, the IZD of Purit, and Hypo was 12 and 14 mm respectively. However, the isolate resisted the effect of the Z germicide and Dettol.
Table 4a and 4b, results of antibacterial activities of antiseptic liquids (Purit, Z germicide, Hypo and Dettol) at different concentration against S. aureus isolated from hospital equipment (stretchers, surgical tables and blood pressure monitors) showed that Purit and Dettol are the most effective antiseptic against the isolated S. aureus. While the isolated S. aureus resisted the effects of Z germicide and Hypo at lower concentration of 12.5.
Also from the result, it was observed that S. aureus isolated from Ventilators, Bed rails and Thermometers were susceptible to Purit and Dettol antiseptics from the highest concentration of 100mg/ml to the least concentration of 12.5 mg/ml. While the isolated S. aureus resisted the effects of Z germicide and hypo at lower concentration of 12.5 mg/ml.
Table 1: Colonial Morphology, biochemical characteristics and percentage occurrence of S. aureus from the skin of medical students
|
Morphology of the isolated S. aureus |
GS |
CT |
COT |
OT |
IT |
POSAAMS % occurrence of S. aureus among male students (n=20) |
POSAAFS % occurrence of S. aureus among female students (n=20) |
TPPISA Total % Prevalence of the Isolated S. aureus (n=40) |
|
Golden yellowish colonies, circular with shiny, smooth surfaces |
+ |
+ |
+ |
_ |
_ |
17 (85%) |
14 (70%) |
31 (77.5%) |
Keys: GS= Gram Stain, CT= Catalase Test, COT= Coagulase Test, OT= Oxidase Test, IT= Indole Test, POSAAMS = % occurrence of S. aureus among male students, + = Positive; - = Negative, POSAAFS = % occurrence of S. aureus among female students, TPPISA = Total % Prevalence of the Isolated S. aureus
Table 2: Colonial Morphology, biochemical characteristics and percentage occurrence of S. aureus from hospital equipment
|
Morphology of the isolated S. aureus |
GS |
CT |
COT |
OT |
IT |
NPSAHS (n=3) |
NPSAST (n=3) |
NPSABPM (n=5) |
NPSAIV (n=3) |
NPSAIBR (n=10) |
NPSAIT (n=10) |
TNPPISA (n=34) |
|
Golden yellowish colonies, circular with shiny, smooth surfaces |
+ |
+ |
+ |
_ |
_ |
3 (100%) |
3 (100%) |
2 (40%) |
2 (66.7%) |
6 (60%) |
4 (40%) |
20 (58.8%) |
Keys: GS= Gram Stain, CT= Catalase Test, COT= Coagulase Test, OT= Oxidase Test, IT= Indole Test, POSAAMS = % occurrence of S. aureus among male students, + = Positive; - = Negative, NPSAIHS = Percentage occurrence of S. aureus isolated from hospital stretchers, NPSAIST= Number and percentage occurrence of S. aureus isolated from surgical tables, NPSAIBPM= Number and percentage occurrence of S. aureus isolated from blood pressure monitors, NPSAIV = Number and percentage occurrence of S. aureus isolated from ventilators, NPSAIBR = Number and percentage occurrence of S. aureus isolated from bed rails, NPSAIT= Number and percentage occurrence of S. aureus isolated from thermometer, NTPPISA = Total number and percentage of the isolated S. aureus.
Table 3: Antibacterial activities of antiseptic liquids (Purit, Z germicide, Hypo and Dettol) at different concentration against S. aureus isolated among male and female medical students.
|
Antiseptic liquids used |
100 mg/ml (mm) |
50 mg/ml (mm) |
25 mg/ml (mm) |
12.5 mg/ml (mm) |
||||
|
|
Male |
Female |
Male |
Female |
Male |
Female |
Male |
Female |
|
Purit |
21 |
18 |
20 |
18 |
15 |
15 |
13 |
12 |
|
Z germicide |
17 |
16 |
15 |
12 |
R |
12 |
R |
R |
|
Hypo |
22 |
20 |
20 |
18 |
17 |
17 |
13 |
14 |
|
Dettol |
20 |
18 |
18 |
15 |
12 |
12 |
12 |
R |
Table 4a: Antibacterial activities of antiseptic liquids (Purit, Z germicide, Hypo and Dettol) at different concentration against S. aureus isolated from hospital equipment (stretchers, surgical tables and blood pressure monitors).
|
Antiseptic liquids used |
100 mg/ml (mm) |
50 mg/ml (mm) |
25 mg/ml (mm) |
12.5 mg/ml (mm) |
||||||||
|
|
HS |
ST |
BPM |
HS |
ST |
BPM |
HS |
ST |
BPM |
HS |
ST |
BPM |
|
Purit |
17 |
16 |
18 |
17 |
14 |
18 |
12 |
12 |
11 |
11 |
10 |
12 |
|
Z germicide |
17 |
15 |
16 |
15 |
13 |
13 |
12 |
R |
12 |
R |
R |
R |
|
Hypo |
18 |
18 |
19 |
18 |
16 |
15 |
14 |
12 |
13 |
11 |
R |
12 |
|
Dettol |
18 |
16 |
18 |
17 |
17 |
18 |
15 |
13 |
13 |
11 |
10 |
11 |
Keys: HS = Hospital stretchers, ST = Surgical Tables, BPM = Blood Pressure Monitors
Table 4b: Antibacterial activities of antiseptic liquids (Purit, Z germicide, Hypo and Dettol) at different concentration against S. aureus isolated from hospital equipment (Ventilators, Bed rails and Thermometers).
|
Antiseptic liquids used |
100 mg/ml (mm) |
50 mg/ml (mm) |
25 mg/ml (mm) |
12.5 mg/ml (mm) |
||||||||
|
|
V |
BR |
TM |
V |
BR |
TM |
V |
BR |
TM |
V |
BR |
TM |
|
Purit |
20 |
19 |
20 |
20 |
17 |
18 |
16 |
17 |
15 |
12 |
11 |
13 |
|
Z germicide |
20 |
18 |
18 |
16 |
15 |
16 |
14 |
12 |
14 |
R |
R |
10 |
|
Hypo |
20 |
18 |
20 |
17 |
16 |
18 |
15 |
14 |
16 |
R |
R |
12 |
|
Dettol |
21 |
18 |
20 |
18 |
17 |
18 |
15 |
15 |
17 |
13 |
10 |
12 |
Keys: V = Ventilators, BR = Bed Rails, TM = Thermometers
DISCUSSION
Staphylococcus species have been identified as the leading cause of Gram-positive bacterial infections and presents wide spectrum of diseases15 both in the hospital and in the community. The results of the analysis of 40 skin swabs, comprising 20 skin swabs from male students and 20 skin swabs from female students, revealed that a total of 31(77.5%) S. aureus prevalence. This percentage prevalence is not unexpected as S. aureus is a normal inhabitant of the skin. Also, the increased daily activities and interaction among these students may also result to the increased prevalence of S. aureus observed in this study. Among the male students, the percentage prevalence of S. aureus was 17(85%), while among female students, the prevalence of S. aureus isolation was 14(70%). The high prevalence of S. aureus observed among male students compared to female students may be attributed to high activities engaged by male students than their female counterparts. An increased activities and interaction between two or more individuals may likely increase the transfer and transmission of resident and pathogenic microorganisms. The most vulnerable part of the body is the epidermis thus, needs adequate protection against adverse environmental conditions and invading microorganisms. Microorganisms can easily colonize the skin surfaces and then spread to other parts of the body, causing various degrees of skin and soft tissue infection. The 31(77.5%) overall prevalence of S. aureus isolation observed in this study amongst the students was found to be higher than 33.34% prevalence of S. aureus isolated from skin of college students reported by Kumar and Mansoor.15 However, they reported 66.66% S. aureus isolation from the anterior nares of the college students.15 In another study conducted by Nwankwo et al., they reported total prevalence rate of S. aureus isolation of 69.5%, with 58(41.7%) amongst male students and 81(58.3%) among female students in a tertiary institution, Abia state, South-Eastern, Nigeria.16 The results of present study and reports of S. aureus in the skin and anterior nares of students went further to establish S. aureus as a leading cause of Gram positive skin and soft tissue infections.17,18 Following the higher isolation of S. aureus amongst male students 17(85%) compared to female students 14(70%), we speculate that the higher prevalence of S. aureus observed in this study amongst male and female students may be due to higher activities engaged by the students.
The result to ascertain the percentage prevalence of S. aureus among frequently used hospital equipment showed the prevalence of 3(100%), 3(100%) and 2(66.7%) for hospital stretchers, surgical tables and ventilators respectively. It was also observed that blood pressure monitors and thermometers with 2(40%) and 4(40%) respectively were the least colonized hospital equipment. The involvement of microorganisms in causing diseases cannot be overemphasized. Therefore, controlling microbial growth has become paramount in hospital environment, industries and at homes in order to inhibit or kill the growth of various pathogenic strains of microorganism. The 100% prevalence of S. aureus observed among hospital stretchers and surgical tables may be as a result of the number of samples collected. Therefore, if more samples were collected, lower prevalence than 100% may be observed. However, the high prevalence of S. aureus observed from these equipment may be justified due to their usage in either carrying or operating patients with complicated health challenges. While the lower prevalence of S. aureus among blood pressure monitor 2(40%) and thermometer 4(40%) may be due to the special care enjoyed by this equipment as most times, thermometers are being cleansed with antiseptics immediately after each usage. For instance, the thermometers are carefully cleaned up before and after usage for each patient. While the blood pressure monitors are carefully and appropriately kept after each use. Most times, this equipment are constantly disinfected using antiseptics or other agents, after each use to make them sterile. However, if not properly disinfected, this hospital equipment can serve as a vehicle for disease transmission.
The results of antibacterial activities of various antiseptic liquids tested among S. aureus isolates recovered from male students showed that Hypo, Purit and Dettol antiseptic liquids were more lethal to S. aureus isolates with inhibition zone diameter (IZD) of 22, 21 and 20 mm at conc of 100 mg/ml and IZD of 13, 13 and 12 mm at the lowest conc of 12.5 mg/ml among Hypo, Purit and Dettol respectively. Also, among the S. aureus isolates recovered from female students, Hypo and Purit with IZD of 20 and 18 mm at conc of 100 mg/ml was the most lethal antiseptics liquid. While at the lowest conc of 12.5 mg/ml, the IZD of 14 and 12 mm are observed among Hypo and Purit respectively. Thus, this results established Hypo and Purit as the most lethal antiseptic liquids against S. aureus isolates compared to Dettol and Z germicide with IZD of 18 and 16 mm at the conc of 100 mg/ml. While, at the lowest conc of 12.5 mg/ml, the S. aureus isolates resisted the effects of Dettol and Z germicide.
Generally, the antibacterial activities of the tested antiseptics against S. aureus isolated from hospital equipment showed to be effective at the highest concentration of 100 mg/ml and 50 mg/ml. While at the low concentration of 12.5 mg/ml, the effect of Z germicide and Hypo reduced. Thus, most of the isolated S. aureus resisted the effect of Hypo and Z germicide. The resistant ability of the isolated S. aureus to Hypo and Z germicide may be due to the increased usage of these antiseptics in both hospital and communities. Hypo, Dettol and Z germicide were mostly used antiseptics at home and hospitals to maintain sterility of home appliances and hospital equipment, thus justifying their reduced efficacy in the study.
From the results, it is therefore imperative to state that the activities of these antiseptics decreased geometrically as the conc of the antiseptic liquids decreased. The result of another study by Rahman et al., also reported Purit to have superior antibacterial activity against S. aureus and E. coli isolates compared to Izal and Hypo.19 However, another study, reported purit and dettol to have a higher activity against S. aureus isolated from water sources compared to Jik and Izal.20 The differences in the ability of these antiseptics to inhibit the growth of S. aureus as observed in this study can be attributed to the differences in the active compound contained by these antiseptics. Also, the difference in the concentrations of the active components may also contribute to the difference in their activity.
CONCLUSION
The antibacterial activities of various antiseptic liquids showed a geometric decrease as the concentrations decreased, suggestive of the activeness of the antiseptic liquids at higher concentration. Thus, the activeness and/or effectiveness of various antiseptics are concentration dependent. Hypo, Purit and Dettol antiseptics were the most active antiseptic liquids compared to Z germicide. Microbial growth in the health care system, industries as well as in the hospitals can be controlled by the use of antiseptic liquids with higher activity. Further studies to ascertain the efficacy of other antiseptics liquids are therefore necessary.
Ethical consideration and Sponsorship:
The products used for this study are only used for advancement of knowledge and deepening scientific knowledge and not in any way intended to be used for any form of litigation or to devalue any of the products. No part of this study was sponsored or financed by any organization, company or institution. The entire study was conceived, sponsored and funded by the authors,
ACKNOWLEDGEMENTS
We appreciate the staff of NOFIC for their assistance in collection of samples from hospital equipment and also the laboratory technologist of Applied Microbiology Department, Ebonyi State University, Abakaliki for their prompt assistance during the analysis.
SOURCE OF FUNDING: This research was entirely funded by the authors
CONFLICT OF INTEREST: The authors declare no conflict of interest
REFERENCES
Ikechukwu Herbert Egwu, M. M. Egwu-Ikechukwu, V. O. Amaechi-Nnaji, O. Nwaeze, Effect of Selected Antiseptic Liquids Sold in Abakaliki Metropolis Against Staphylococcus Aureus Isolated from Medical Students and Hospital Equipment in A Tertiary Care Hospital, Abakaliki, Ebonyi State., Int. J. of Pharm. Sci., 2025, Vol 3, Issue 7, 3594-3603. https://doi.org/10.5281/zenodo.16447543
10.5281/zenodo.16447543
