Survey the Frequency of Bacterial Species Isolated from Smartphones among College Students 2023 A.D.

Ministry of Higher and Scientific Research University of Anbar College of Pharmacy A research submitted in partial fulfilment of the requirements for Bachelor of Science Degree in Pharmacy By Saja Nazar Mahmoud...

Ministry of Higher and Scientific Research University of Anbar College of Pharmacy A research submitted in partial fulfilment of the requirements for Bachelor of Science Degree in Pharmacy By Saja Nazar Mahmoud Ghufran Anwer Hameed Safa Safaa Abdulqader Nuha Hamid Khaled Teba Feras Madallh Mohammed Saad Turki Supervised by Dr. Sulaiman Ajaj Abdullah 2023 A.D. 1444 A.H. ‫سورة اجملادلة – اآلية ‪11‬‬ ‫‪I‬‬  Acknowledgement First of all, Praise be to God Almighty who has helped us to complete this scientific research, and who has given us health, wellness and determination. We extend our thanks and appreciation to our supervisor, Dr. Sulaiman AL-Qaesi for all scientific guidance, and valuable information he provided us that contributed to enriching the subject of our study in its various aspects. Our profound thanks to Prof. Dr. Zeina Al-Sabti the deanery of the college of Pharmacy, University of Anbar; Assist. Prof. Dr. Atheer Khalaf and Assist. Prof. Dr. Esraa Adnan representatives of the Deanship for all their support. We also wish to express our grateful thanks to Dr. Rawaa for her assistance during the practical experiments. We are deeply grateful to our teachers and our colleagues in the college for their support during our study years. Our deepest appreciation is directed to the participating students who expressed their assistance and made this work possible. II  Abstract Background: Mobile phones (MPs) have emerged as one of the most essential accessories in social and business activity. Despite their numerous advantages, MPs provide a lot of opportunities for infectious diseases to spread throughout communities. This study aimed to determine the microbial species associated with the contamination of mobile phones of certain University of Anbar / College of Pharmacy students, in order to take the necessary precautions. Methods: Along with the questionnaire, a swab sample from 40 participant’s MPs were collected. Sterile swabs were passed over the exposed surfaces of the MPs, then inoculated onto blood agar and MacConkey agar plates. The bacterial species identified microscopically and macroscopically as well as by biochemical tests. Results: The percentage of bacterial contamination on individual MPs was 80%. Escherichia coli was the most common bacterium isolated (38.235%), followed by Staphylococcus aureus (32.353%), Klebsiella pneumonia (17.647%), Pseudomonas aeruginosa (8.823%), and Proteus vulgaris (2.941%). Staphylococcus aureus was the most common bacterium in males, according to an assessment of bacterial type and incidence by gender, while E. coli was the most common bacterium in females. Klebsiella pneumonia was more common in the first stage than the fifth stage, while Staphylococcus aureus and E. coli were more common in the fifth stage than the first stage. Conclusions: According to this study, most of the mobile phones under consideration were contaminated with various microbes, the majority of which belonged to the normal human body flora. To reduce the risk of cross-contamination and healthcare-associated infections carried by MPs, comprehensive recommendations regarding limiting the use of MPs in laboratory settings, hand hygiene, and frequent decontamination of MPs are advised. III  List of contents Item no. Title Page no. 1 Introduction 1 2 Material and methods 3 2.1 Samples Collection 3 2.2 Bacterial Culturing and Identification 3 2.3 Biochemical tests 4 2.3.1 Coagulase test 4 2.3.2 Oxidase test 5 2.3.3 IMVC test 5 3 Results 6 3.1 Isolation of Bacteria 6 3.2 Identification of Bacteria 6 4 Discussion 12 5 Conclusion 15 6 Future prospects 15 References 16 Appendix 18 IV  List of tables Table Page Title no. no. Bacteria isolated from mobile phones in University of Anbar / 1 10 College of pharmacy Comparison of bacteria isolated from MPs of male and female at 2 11 University of Anbar / college of pharmacy Comparison of bacteria isolated from MPs of first stage and fifth 3 11 stage at University of Anbar / college of pharmacy 4 A summary of the results obtained from the questionnaire 19 List of figures Figure Page Title no. no. The frequency of bacterial contamination among the swabbed 1 6 phones. 2 Bacterial isolates on blood agar plates. 7 3 Bacterial isolates on MacConkey agar plates. 7 IMVC tests for bacterial isolates. (A) IMVC results for E. coli, 4 (B) IMVC results for Klebsiella pneumonia and (C) IMVC 8 results for Proteus vulgaris Coagulase tests to identify Staphylococcus aureus, (A) coagulase 5 9 +ve, (B) coagulase tests -ve Oxidase tests to identify Pseudomonas aeruginosa, (A) oxidase - 6 9 ve, (B) oxidase tests +ve The growth of Staphylococcus aureus on blood agar medium 7 10 showed β hemolysis V  1. Introduction A mobile phone (MP) is a portable electronic device with a long range that is used for long-distance personal communication. MPs have evolved into a necessary tool for social and professional life due to recent advancements in information sources and social media apps. As the future of higher education appears to be technology-driven, the importance of technology has significantly risen over the last few years in assisting students' learning processes. However, one of the most frequent worries about the extensive usage of MPs is that they might serve as a means of spreading harmful germs and other pathogens. Due to the development and advantages of MPs, their risk to human health is frequently disregarded. The continual handling, heat generated by phones, and sweat from hands provide a perfect breeding environment for all types of germs that are typically found on the skin. Through repeated hand contact, a cell phone might spread contagious diseases. Tens of thousands of microorganisms reside on each square inch of mobile phones, making them a potential health risk. Staphylococcus aureus is widespread bacteria that can be found on the skin and in the noses of up to 25% of healthy people and animals. It can cause infections like boils and pimples as well as pneumonia and meningitis and is a close relative of methicillin resistant Staphylococcus aureus (MRSA). This is because the isolated bacteria constitute a subset of the skin's typical microbiota, according to earlier studies [5, 6]. Even while the microbes that health researchers have so far isolated are generally a part of the normal flora of the site of contamination, they can nevertheless lead to opportunistic diseases. Studies have shown that sources such as human skin or hands, phone pouches, bags, pockets, the environment, and food particles can contaminate mobile phones. These 1 sources are connections through which microorganisms colonized the phone, leading to diseases that can range from mild to chronic. Aims of this study 1. Isolation of bacterial flora contaminated a mobile phones of certain University of Anbar / College of Pharmacy students. 2. Characterization and identification of isolated bacteria. 3. Check to see if these mobile phones offer any significant health risks and look into personal hygiene practices. 2 2. Materials and methods 2.1 Samples Collection The samples were collected and examined in accordance with the method of streak plate technique. Forty cell phones belonging to 40 students (20 males and 20 females) from at University of Anbar / College of Pharmacy were screened. Pre-tested questionnaire was filled out by each participant and inquired about stage, gender, health status, use of headphone, the duration of phone ownership and routinely phone cleaning. Samples were taken aseptically by rolling a sterile cotton swabs soaked with sterile normal saline over the exposed surfaces of the MPs. Extreme caution was applied to ensure that the screen, side of the phone, switches, ear socket and audio input were sufficiently wiped given how frequently the user uses their hands and fingers to access these areas. For MPs with covers, in addition to the screen, the swab was obtained from the outer surfaces of the cover. 2.2 Bacterial Culturing and Identification The collected swabs were placed immediately in a sterile container and transferred in less than 30 minutes to the Microbiology Laboratory. Following the conventional streak plate procedure, the obtained samples were inoculated onto blood agar and MacConkey agar plates. The plates were then incubated aerobically at 37 °C for the following 24-48 hours. Based on colony characteristics and microscopic examination of bacterial smears stained with Gram stain, the initial identification of bacteria was done. For further identification, various biochemical assays including indole, methyl red, Voges-Proskauer, citrate, catalase, coagulase, and oxidase were used. 3  2.3 Biochemical tests Biochemical tests performed to confirm the diagnosis of bacteria. S. aureus was subjected to a coagulase test, Pseudomonas was identified using an oxidase test, while IMViC tests were used to differentiate between members of the Enterobacteriaceae family. 2.3.1 Coagulase test Staphylococcus aureus strains are distinguished from other coagulase-negative organisms using the coagulase test. Coagulase is an enzyme-like protein that causes plasma to clot by converting fibrinogen to fibrin. Staphylococcus aureus produces two forms of coagulase: free coagulase and bound coagulase. Method: Two test tubes, one of which is labeled with the strain number to be tested and the other with a negative control. Each test tube received 0.5 mL of the reconstituted plasma using a pipette. Two or three isolated bacterial colonies were collected using a sterile loop or applicator stick. The bacteria emulsified in the 0.5 ml of plasma tube. The control tube left without inoculation. The two tubes placed in the incubator. Over the course of the following four hours, the culture was periodically checked for the existence of a clot. The test can be continued if no clots are seen after 4 hours with an overnight incubation at room temperature and a final observation at 24 hours. 4  2.3.2 Oxidase test The oxidase test is essential for distinguishing between the Pseudomonadaceae (ox +) and Enterobacteriaceae (ox -) families of bacteria. It is also helpful for identification and speciation a wide variety of other bacteria, including those that use oxygen as the final electron acceptor during aerobic respiration. Method: A freshly prepared tertramethyl-p-phenylene-diamine dihydrochloride (1%) solution is added to a strip of Whatman's No. 1 filter paper. With a platinum loop, the test colony is picked up and applied to the moist region. Appearance of deep-purple color within 5 to 60 seconds of exposure indicates a positive reaction, while a lack of indicates a negative response. 2.3.3 IMVC tests The IMVC series is a collection of four distinct tests, usually used to detect bacterial species, particularly coliforms. Each of the four tests is represented by a capital letter in the acronym IMVC: I for Indole test, M for Methyl Red test, V for Voges-Proskauer test, and C for Citrate test. Method: Every tested strain of bacteria was inoculated into four culture tubes. One contains a peptone water medium for the indole test, the other two contain MRVP media for the methyl red and Voges-Proskauer tests, and the final tubes contains a Simon-citrate medium for the citrate test. The tubes incubated in 37oC for 24hrs. Results were reported following the incubation time. 5  3. Results 3.1 Isolation of Bacteria A total of 40 students (20 males and 20 female) from first and fifth stages College of Pharmacy / University of Anbar participated in this study. The frequency of bacterial contamination among the swabbed phones were 32 of 40 (80%); are shown in (figure 1). Tested phones (40) Contaminated phones 20% Non contaminated phones 80% Figure 1: The frequency of bacterial contamination among the swabbed phones. 3.2 Identification of Bacteria Bacterial isolates were recognized macroscopically according to colony characteristics on blood agar plates (figure 2) and on MacConkey agar plates (figure 3) and microscopically regarding Gram staining. 6  Figure 2: Bacterial isolates on blood agar plates. Figure 3: Bacterial isolates on MacConkey agar plates. 7  The identification of isolated bacteria confirmed using biochemical tests. IMVC tests done for gram negative bacteria belong to Enterobacteriaceae family. Majority of these bacteria detected as being E. coli were (Indole +ve, Methyl red +ve, Voges-Proskauer -ve and citrate –ve). Other types of bacteria were (Indole -ve, Methyl red -ve, Voges- Proskauer +ve and citrate +ve) suggested contamination with Klebsiella pneumonia. One variety of bacteria showed contamination with Proteus vulgaris (Indole +ve, Methyl red +ve, Voges-Proskauer -ve and citrate +ve) (figures 4). Figure 4: IMVC tests for bacterial isolates. (A) IMVC results for E. coli, (B) IMVC results for Klebsiella pneumonia and (C) IMVC results for Proteus vulgaris 8  Coagulase tests done for bacterial isolates that detected as being Staphylococcus, revealed coagulase positive Staphylococcus aureus (figure 5). A B Figure 5: Coagulase tests to identify Staphylococcus aureus, (A) coagulase +ve, (B) coagulase tests -ve Oxidase tests done for bacterial isolates that detected as being Pseudomonas aeruginosa showed positive results (figure 6). Figure 6: Oxidase tests to identify Pseudomonas aeruginosa, (A) oxidase -ve, (B) oxidase tests +ve Gram-negative bacteria made up the majority of these bacteria (67.65%). Escherichia coli accounted for 38.235% of these bacterial samples, followed by Klebsiella pneumonia (17.647%), Pseudomonas aeruginosa (8.823%), and Proteus vulgaris (2.941%). In terms of gram- 9 positive bacterial isolate, coagulase positive Staphylococcus aureus accounted for (32.353%) (table 1). Table 1: Bacteria isolated from mobile phones in University of Anbar / College of pharmacy Bacterial isolates Number Percentage Escherichia coli 13 38.235% Staphylococcus aureus 11 32.353% Klebsiella pneumonia 6 17.647% Pseudomonas aeruginosa 3 8.823% Proteus vulgaris 1 2.941% Total no. 34 100% The growth of Staphylococcus aureus on blood agar medium plainly displayed β hemolysis with clear zone around the colonies (figure 7). Figure 7: The growth of Staphylococcus aureus on blood agar medium showed β hemolysis Bacterial isolates contaminating MPs of both males and females were observed as reflected in (table 2). A comparison of bacterial types and frequency among both groups showed that Staphylococcus aureus was 10 the most prevalent bacterium in males, whereas E. coli was the most prevalent bacterium in females. Table 2. Comparison of bacteria isolated from MPs of male and female at University of Anbar / college of pharmacy Male Female Type of bacteria No. (%) No. (%) Escherichia coli 5 (38.46%) 8 (61.54%) Staphylococcus aureus 6 (54.54%) 5 (45.45%) Klebsiella pneumonia 3 (50%) 3 (50%) Pseudomonas aeruginosa 1 (33.33%) 2 (66.66%) Proteus vulgaris 1 (100%) 0 Total no. 16 18 As shown in (table 3), bacterial strains were found to be contaminating first- and fifth-stage MPs. According to a study of the types and frequencies of the bacteria in the two groups, Klebsiella pneumonia was more common in the first stage than the fifth stage, while Staphylococcus aureus and E. coli were more common in the fifth stage than the first stage. Table 3. Comparison of bacteria isolated from MPs of first stage and fifth stage at University of Anbar / college of pharmacy First stage Fifth stage Bacterial isolates No. (%) No. (%) Escherichia coli 5 (38.46%) 8 (61.54%) Staphylococcus aureus 3 (27.27%) 8 (72.73%) Klebsiella pneumonia 5 (83.33%) 1 (16.67%) Pseudomonas aeruginosa 2 (66.66%) 1 (33.33%) Proteus vulgaris 1 (100%) 0 Total no. 16 18 11 4. Discussion Mobile phones are now among the most important tools for both social and professional life. Mobile phones are frequently handled and held near to the face, despite the fact that they are typically kept in bags or pockets. However, the use of MPs by students at the College of Pharmacy / University of Anbar raises questions about sanitation and the ready spread of bacteria that could have serious negative effects on health. The results showed that 80% of MPs examined in our study were contaminated with different types of bacteria. These results comparable to those of Auhim, M.A. who observed that the rate of bacterial contamination of personal mobile phones was 82.5%. They are contrast with those of Selim et al. who state higher rate of bacterial contamination in mobile phones 100%, as well as Adhikari et al. in Nepal, who found a lower prevalence of bacterial contamination (56%). The observed variation could be attributed to differences in infection prevention or frequency of cleaning MPs, hand washing practice, and public awareness of the role of an MP in microbial transmission. Escherichia coli, staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumonia and Proteus vulgaris are the main bacterial isolates frequently associated with mobile phones of our research as shown in (table 1). According to the findings in (table 2) above, staphylococcus aureus and E coli were the two most common bacteria found in cell phones belonging to males and females, respectively. The results demonstrate that staphylococcus aureus and E coli are the two major bacterial isolates frequently linked to student-owned mobile phones. The risk of using a mobile phone in a hospital has also been highlighted by this research. Our study's 12 findings in (table 3) led us to the conclusion that the fifth stage had a higher percentage of E. coli and Staphylococcus aureus than the first stage, possibly because the student receiving hospital training, which may have been the cause of the contamination. All of the organisms isolated in this study are opportunistic pathogens. They do not represent a threat under normal circumstances, but patients with compromised immune systems may be at risk. These microorganisms most likely got their way into the phone through the skin and passed from hand to hand. This is due to the fact that the isolated bacteria are a subset of the established normal microbiota in human. These microbes, though, have numerous adverse health consequences. For example, Staphylococcus aureus is associated with a variety of diseases, from boils and pimples to pneumonia and meningitis; this idea is supported by the large number of colony isolates. Similarly, although E. coli is a normal microbiota of human GIT, it has developed pathogenic processes that allow it to cause diseases. They can lead to enteritis or extra-intestinal infections, most frequently a urinary tract infection. According to London School of Hygiene & Tropical Medicine, 28% of the devices were contaminated with E. coli, which can result in food poisoning. This is most likely because people don't properly cleanse their hands after using the restroom. K. pneumoniae is readily colonizes human mucosal surfaces, from there, it enters other organs and spreads, leading to serious infections like pneumonia, UTI, and liver abscesses. Although Pseudomonas aeruginosa is a part of the normal flora, it is extremely dangerous to human health due to their ability to create biofilm. Proteus vulgaris has also been associated to respiratory infections, burns infections, wound infections, and UTI. Fecal 13 bacteria, particularly in warmer temperatures away from sunlight, can survive on hands and surfaces for hours. Touching things like food, door knobs, and even MPs makes it easy to spread. MPs were becoming real reservoirs of pathogenic agents as they contact ears, lips, faces and hands of different users with varying health conditions. Microbial infections could be reduced by identifying and controlling risk factors, educating people, and conducting microbial surveillance. Most people are unaware of the dangers of phone sharing. Undoubtedly, sharing phones entails cross-sharing. Effective disinfecting methods should be developed to reduce the potential biological hazards of cell phones. Because restricting the use of mobile phones is not an actual solution, many researchers advocate for strict adherence to infection control precautions such as hand hygiene. Furthermore, people should be made aware that MPs may be a source of hospital-acquired infections being transmitted to and from the community. 14 5. Conclusion Cell phone usage has extremely increased; nearly everyone has one in their pocket, but they are unaware that their MPs are inhabited with microorganisms. People are insensible of this and use them in a variety of settings, including hospitals, while eating, and even while using the restroom. Our findings suggest that these cell phones can serve as mobile carriers for both pathogenic and non-pathogenic microbes. Individuals ought to be informed of the risks associated with mobile phones and urged to care of disinfecting their devices in order to reduce the risk of contracting infections while handling them. They should also be made aware of the dangers of using mobile phones in confined spaces. 6. Future prospects More research is needed to evaluate the effectiveness of the above mentioned methods and identify more effective substitutes for reducing bacterial contamination and preventing the spread of infections through mobile phone use. 15  References 1. Borth, D.E. Mobile telephone. 2023; Available from: https://www.britannica.com/technology/mobile-telephone 2. Bahja, M., M.A. Kuhail, and R. Hammad, Embracing Technological Change in Higher Education. Higher Education-New Approaches to Accreditation, Digitalization, and Globalization in the Age of Covid, 2021. 3. 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Microbiology and molecular biology reviews, 2016. 80(3): p. 629-661. 20. Tuon, F.F., et al., Pathogenesis of the Pseudomonas aeruginosa biofilm: A review. Pathogens, 2022. 11(3): p. 300. 21. Ahmed, D.A., Prevalence of Proteus spp. in some hospitals in Baghdad City. Iraqi Journal of Science, 2015. 56(1): p. 665-672. 22. Tarimo, I.A., Environmental Sanitary Conditions of Primary Schools in Lindi Municipality Tanzania. 23. Panigrahi, S.K., et al., Covid-19 and mobile phone hygiene in healthcare settings. BMJ global health, 2020. 5(4): p. e002505. 17  Appendix The survey's inquiry form User Survey Questionnaire of the Medicines Information Service Title: Survey the frequency of bacterial species isolated from smartphones among college students. No. Gender : Query Response 1. Health status 2. Use of headphone 3. The duration of phone ownership 4. Do you routinely clean your phone? 18  Table 4: A summary of the results obtained from the questionnaire Status Variants No. (%) Normal 15 (75%) Health status Sick 5 (25%) Yes 17 (85%) Use of headphone No 3 (15%) Less than 1 year 7 (35%) Duration of phone ownership More than 1 year 13 (65%) yes 12 (60%) Routinely clean the phone No 8 (40%) 19