A STUDY ON THE ISOLATION AND IDENTIFICATION OF AIR MICROFLORA IN MICROBIOLOGY LABORATORY

ABSTRACT:

This study was conducted to isolate and identify bacteria air pathogens in our college lecture rooms using the natural sedimentation technique. The studied areas include political science lecture room III, criminology and security study lecture room III, English lecture room III, environmental management and toxicology lecture room II, mathematics lecture room II, physics lecture room II, postgraduate lecture room and main hall science. A total of 64 samples were collected during the course of this study out of which 56 (87.5 %) showed positive bacterial growth. The isolated bacterial species were identified as Staphylococcus aureus, E. coli, Streptococcus species and Bacillus subtilis. Staphylococcus aureus had the highest percentage occurrence of 51% followed by E. coli (25%) and Streptococcus species (21%) while Bacillus subtilis recorded the least (3%). These pathogens could be linked with several infections such as gastrointestinal tract infections, respiratory tract infections, urinary tract infections and skin disorders. These findings would alert the students, staff and workers to these pathogens and their existence in our lecture rooms.

CHAPTER ONE INTRODUCTION

1.1 Background of the Study

Microorganisms are found almost everywhere, and their presence in the air was demonstrated by the work of Lazzaro Splallanzani in 1768 and of Louis Pasteur at the end of the 19th century (Meraj-ul-Haque et al., 2016). However, air is not a natural medium for growth and reproduction of microorganisms, any organism, that airborne contain must have originated from a living or non living source (humans, animals, plants, food, water or soil) (Yaghoub and Elagbash, 2010). Though microorganisms are found in both indoor and outdoor environments, most of the people spend their lives indoors: in houses, industries, offices, colleges, schools, hospitals etc., where they are exposed to many bioaerosols (biological air borne contaminants such as bacteria, viruses, fungi or their byproducts). Exposure to these airborne particles can result in respiratory disorders and other adverse health effects such as infections, hypersensitivity pneumonitis and toxic reactions (Naruka and Gaur 2014; Sheik et al., 2015; Yassin and Almouqat, 2010). In addition, long-term contact of people with bioaerosols can influence a person’s mental power and learning ability (Naruka and Gaur, 2014). Different environmental conditions such as temperature, UV light, dryness and humidity, play a role in controlling the growth of airborne particles. Nevertheless the microbes manage to reach new hosts through the air for its survival (Sheik et al., 2015). Poor ventilation, crowded conditions and increase in number of air conditions inside building nowadays can facilitate the spreading and the survival rates of airborne particles and also can increase the chance of people at risk of airborne infections. Among dust particles present in the indoor environment, fungus which reproduce by forming spores, some bacteria especially gram positive bacteria and some viruses can survive for a long time in the air (Sheik et al., 2015; Jacob, 2016). Despite the need to monitor bio-aerosol levels in evaluating health risks, differences between automatic techniques and passive sedimentation techniques hamper results comparison. Automated techniques, although they are efficient in quantitative analysis, are of limited use because they require heavy and noisy equipment and need a constant power supply. The passive sedimentation technique is also limited because it does not permit an adequate quantitative analysis, but it is still widely recommended in the literature for use as a microbiological alert (Abe et al., 2012). It is well known that exposure to bioaerosols can cause adverse effect on people and for this reason it is important to check the sanitary conditions of air in the place they live. This research work aims to isolate and identify bacteria in the air of Federal University Dutse lecture rooms, to the best of our knowledge no such study has been attempted till now.

1.2 Materials and Methods

 Sampling Site This study was carried out at federal university Dutse. The samples were collected from eight different lecture rooms namely, Political Science lecture room III, CSC lecture room III, English lecture room III, EMT lecture room II, Maths lecture room II, Physics lecture II, PG Lecture room and MHS. Media Preparation The media used were prepared according to the manufacturer’s instructions. Sample Collection Sedimentation Technique which involves the opening of plate with specific culture media was employed for this study (Sekulska, 2007). Prepared plates of nutrient agar were exposed to air for 30mins at different sites in the respective lecture rooms. After sampling, all plates were immediately taken to the microbiology laboratory and incubated at 37ºC for 24 hours for isolation of bacteria. The colonies were sub-cultured onto a new fresh medium in order to obtain pure culture. ______________________  Musa Hassan Muhammad, Fiddausi Uba, Madinat· R., Muhammad Muhd, M.A Sulaiman, Munzali S. Zubair  Federal University Dutse, Department of· Microbiology and biotechnology  Phone Number:+ 2347068823187·  E-Mail: musachakla@yahoo.com· INTERNATIONAL JOURNAL OF SCIENTIFIC & TECHNOLOGY RESEARCH VOLUME 6, ISSUE 09, SEPTEMBER 2017 ISSN 2277-8616 17 IJSTR©2017 www.ijstr.org Bacterial Identification Identification of bacterial isolates was done using the standard procedures according to Cheesbrough (2009). Bacterial colonies were initially characterized by morphology and using staining techniques (Gram staining) and identified further by biochemical tests. Gram Staining Gram’s staining was done to find the reactions of the bacterial isolates to Gram reagents. A smear was prepared and heat fixed. The crystal violet (primary stain) stain was flooded over the fixed culture for 60 seconds, the stain was washed with water. The iodine solution was added onto the smear for 60 seconds, pour off and rinsed with water. A few drops of decolorizer (ethyl alcohol) was added and washed with water immediately after 5 seconds and finally safaranin (Secondary stain) was added for 60 seconds and washed, the smear was allowed to air dry. After drying the slide was mounted under microscope and observed. The stain differentiates bacterial species into two groups; Grampositive bacteria, which take up crystal violet dye (primary stain) and are stained violet and Gram-negative, which pick up safranin (Secondary stain) are thus stained red after decolourization with alcohol. Biochemical Test Biochemical tests such as Catalase, Oxidase, Indole, Methyl Red test (MR), Coagulase, Voges Proskauer (VP), Citrate utilization, were carried out on the isolated bacteria according to cheesbrough (2009). Result and Discussion The present study was conducted to isolate and identify airborne bacteria in some selected lecture rooms within the Federal University Dutse. A total of sixty four samples were collected during the course of this study out of which fifty six (87.5 %) were positive while eight (12.5%) showed negative bacterial growth (Table 1). Four bacterial genera were identified from the sampling sites as shown in Table 2. Overall, 124 bacteria were isolated comprising of Staphylococcus aureus, Streptococcus spp, E. coli and Bacillus subtilis (Table 3). Staphylococcus aureus has the highest percentage occurrence of 51% followed by E. coli (25%) and Streptococcus species 21% while Bacillus subtilis recorded the least 3% (Table 3). These pathogens could be linked with several infections such as gastrointestinal tract, respiratory tract, urinary tract and skin disorders.

1.3 Result and Discussion

 The present study was conducted to isolate and identify airborne bacteria in some selected lecture rooms within the Federal University Dutse. A total of sixty four samples were collected during the course of this study out of which fifty six (87.5 %) were positive while eight (12.5%) showed negative bacterial growth (Table 1). Four bacterial genera were identified from the sampling sites as shown in Table 2. Overall, 124 bacteria were isolated comprising of Staphylococcus aureus, Streptococcus spp, E. coli and Bacillus subtilis (Table 3). Staphylococcus aureus has the highest percentage occurrence of 51% followed by E. coli (25%) and Streptococcus species 21% while Bacillus subtilis recorded the least 3% (Table 3). These pathogens could be linked with several infections such as gastrointestinal tract, respiratory tract, urinary tract and skin disorders.

Conclusion

 Using the natural sedimentation technique, 124 bacterial species were isolated in this study. Four genera of the organisms have been isolated comprising of Staphylococcus aureus, Streptococcus spp, E. coli and Bacillus subtilis. And these organisms can cause several infections to the students and staff. In order to develop the quality of indoor air in our lecture room buildings overcrowding has to be avoided, good ventilation systems has to be designed and good hygiene practice must be observed. Recommendations Based on the findings of this study, we recommend the followings: (i) Proper ventilation system should be provided when constructing lecture rooms to allow for movement of air in and out of the building. (ii) To protect the health of students and staff proper control measures has to be taken to prevent the environmental factors which facilitate the growth and proliferation of pathogenic bacteria in our lecture rooms. (iii) Disinfection of floors should be performed routinely and dust should be prevented (iv) Finally, this study focus on the isolation of bacterial airborne pathogens, Future studies may be required to isolate fungal pathogens which could be a good source of air borne pathogens.

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