Assessment of the air quality of some selected classroom surfaces in the south campus of federal
polytechnic ede.
Introduction
Air pollution is the most serious environmental health threat on the planet. Pollution is responsible for
one out of every eight deaths worldwide. According to reports, 7 million people died in 2012 as a result
of exposure to ambient and indoor air pollution. In 2012, Africa accounted for over 680,000 of the total
global air pollution-related deaths (WHO).Over the years, scientific study has linked bioaerosol exposure
to the incidence of undesirable health impacts such as communicable infectious diseases, acute toxic
effects, allergies, and cancer.Liu et al., (2018) investigated the total numbers of harmful bacteria in the
air in Hangzhou (China). 1
During the experiment,Airsampling which included dust of various particle sizes and quality indexes,
and 16SrRNA sequencing was used to characterise the bacteria present. Thiobacillus, Methylobacterium,
Rubellimicrobium, and Paracoccus, all belonging to the Proteobactria genus, were found to be the most
numerous. Staphylococcus, Bacillus, Clostridium, Enterobacter, and Klebsiella were the most common
pathogenic bacteria generally found in airborne particulate matter (PM) samples.Kubra et al. (2015)
conducted microbiological air quality tests in selected kindergartens. In the morning and afternoon, air
samples were obtained in classrooms.
The quality of indoor air was analysed using Polish standards (PN-Z-04111-01; PN-Z-04111-02:1989; PN-
Z-04111-03:1989) based on groups of indicator microorganism gathered outside the kindergartens as a
control sample. The total quantity of 1heterotrophic bacteria and staphylococci in the air surpassed
allowed levels regardless of the research method used, while no significant contamination with mould
fungi was discovered.Nabrdalik I Latala (2003) conducted study to characterise the quantitative and
qualitative characteristics of fungi found in building structures. They revealed that, due to the presence
of potentially dangerous fungi in the examined objects, it is required to develop testing techniques and
criteria to determine the degree of air pollution in residential structures.Pastuszka et al.(2000)
conducted research to evaluate the concentrations of bacterial and fungal bioaerosols in various types
of buildings, as well as the risk of specific diseases caused by these bioaerosols. They discovered that
there was an increased incidence of asthma symptoms in buildings with mould issues. Inhalation of
fungal particles and other airborne substances most likely raised the risk. The relative concentration of
the observed species, including Penicillium species, ranged from 3 to nearly 50% of the fungal
population in the house without mould, but the highest concentration of Penicillium species was
identified in 90% of all fungi in the indoor air. Indoor and outdoor airs, as well as atmospheric air, are all
important factors in the normal functioning of the human body. Air pollution is one of the most serious
risks to people's living surroundings. Environmental contaminants are potentially dangerous airborne
pollutants that substantially impact human health, according to the World Health Organization (WHO)
and the European Environment Agency (EEA). All compounds in the earth's atmosphere are not natural
components, greatly elevated quantities of such compounds in air signifies air pollution. Chemical
compounds, acid rain, airborne ashes, dust, trace elements and biological contaminants the
atmosphere.Pollen, bacteria, and viruses are examples of biological air pollutants, often known as bio
aerosols. Microorganisms pollute the air, plant surfaces, rocks, buildings, and hospital environments.
The dispersed phase of the bio aerosol is made up of particles with sizes ranging from 1 to 200 µm.
Some may be pathogens or spread allergies, putting the population's health at risk.Microorganisms, as
we all know, are one of the most important variables impacting human health. Microorganisms can be
found under a wide range of conditions, including severe temperatures, pressures, salinity, and acidity.
One of the remaining biological limits on Earth has been identified as the atmosphere. The microbial
community's composition and biodiversity in the atmosphere are still poorly understood. Bacteria and
fungi have been found in several atmospheric layers, including the boundary layer, upper troposphere,
and stratosphere at altitudes of more than 20 kilometres above sea level. Airborne microorganisms have
the ability to travel long distances, which has an impact on air quality.Microorganisms present in
outdoor and indoor air impacts human health through residential, hospital, and public spaces.They can
be dispersed through the air by wind and precipitation, resulting in deterioration of air quality. As a
result, the presence of microorganisms in both the ambient air and the indoor environment,particularly
those that cause infectious diseases, can be extremely harmful. Microorganisms in the air have been
shown to be responsible for immune system anomalies such as allergies, infections, airborne diseases
and even cancer.The purpose of this research article is to present up-to-date facts on the assessment of
the air quality of some selected classroom surfaces in the south campus of federal polytechnic ede.
Many studies by researchers established a correlated between the air quality and human cognitive
performance. However, the chosen study area has not been much studied so far. During the COVID- 19
pandemic situation, the Government of India declared a complete lockdown from March 2020. A
prelockdown, and lockdown situation was closely analysed and monitored to find out whether limited
transportation and human activity outdoor plays any major role in reducing number of pathogens. The
outdoor air quality also impacts indoor air quality. Hence, both outdoor and indoor air quality
assessment was done considering the data on bio- aerosolised particles. The study may reveal the
impact of road transportation and air pollution on altering the air quality. Changes in terms of biological
air quality, was significantly studied. Air quality has an impact on the immunological development and
the emergence of allergies, asthma, and allergic diseases during early childhood is evident.Bio aerosols
in the air could be a source of physiological ailment. Asthma, hay fever, bronchitis, chronic lung failure,
lung cancer, and other diseases are all caused by airborne germs, particularly bacteria and fungi. Aside
from pathogenic microorganisms in the air, attention should also be paid on the biologically active
products, such as endotoxins or mycotoxins; these airborne biological agents and their toxic
components are major issues related to human health and development.
The main objectives of the present study are as follows:
1. To isolate bacteria, fungi from air (outdoor and indoor) using culture dependent technique and plate
exposure method.
2. To enumerate the number of bacteria and fungi present in air.
3. To study the colony morphology of typical colonies obtained by plate exposure method.
4. To identify selected colonies by staining and microscopy.
5. To perform biochemical identification and type study of the selected colonies.
6. To perform statistical calculations for quantifying the number of microbial pollutants present in air3
7. Lastly, interpretation of the microbial air quality of the present study area.
This study was conducted to enumerate the numbers of living microbes or bio aerosols suspended in the
air. It was intended to check the outdoor and indoor microbial quality of air. As the microorganisms are
directly involved in affecting human health and the environment. The scope of work includes some
selected classroom surfaces in the south campus of federal polytechnic ede to assess the air quality of
this selected classroom surfaces .
Statement of problem
The following are the causes responsible for low air quality
-Induatrial emission : Emission of large quantities such as Sulfur dioxide(so2), nitrogen
oxide(NOx),particulate matter(pm) and volatile organic compounds(VOCs) due to combustion of fossil
fuels and other industrial processes can lead to low air quality and cause air pollution.
-vehicular emission: Exhaust fumes from cars , trucks and buses emit carbon monoxide (CO), NOx, Pm,
and hydrocarbon from the combustion of gasoline and diesel causes air pollution.
-Agricultural activities : Application of chemical in farming releases ammonia and other pollutants into
the air which causes air pollution.
-Natural sources : Natural occurrences such as volcanic eruptions ,dust storms and wildfires produces
pollutants such as carbon monoxide ,sulfur dioxide ,dust which significantly impacts air quality .
-Household Activities: Activities such as smoking , cooking , cleaning agents and paints can produce
poisonous and harmful emission which can greatly impact the air quality of a certain environment.
-Overcrowding : Places with limited space can cause congestion of occupants which can lead to
production of unhealthy gases and smokes , sweats which thereby lead to impacts on air quality of a
certain place at a particular time.
3 Methods and Materials
The following ways are methods in which the assessment of air quality of some selected classroom
surfaces in the south campus of the federal polytechnic ede can be conducted.
2.1 Experimental Setup
Extensive microbiological culture dependent analysis was conducted for examination of the air quality.
Air exposure method/ plate settling method, quantification of number of bio-pollutants, and grading the
air qualitywere the main aim of the experimental study. The experimental setup included:
1. Selection of the study area: IPHH hospital and laboratory (Indoor) and IPHH terrace and lawn and the
campus area (Outdoor).
2. Extensive microbiological culture dependent analysis was carried out. Air exposure method/ plate
settling method were performed.
3. Statistical enumeration: Arithmetic mean and standard deviation of the parameters were calculated
using Microsoft Excel 2003and student’s one sample t- test was calculated using SPSS version 26
software. Various methods were applied for enumeration and detection of microorganisms in air which
are as follows: sedimentation methods, microscopic methods and culture methods.
2.1.1 Sedimentation method
The easiest method is the 'Settling Plate Technique,' which is extensively used by air microbiologists. In
this experimental study bacteria-carrying particles are allowed to settle onto the medium for a length of
time before being incubated at the proper temperature. The quantity of settled bacterium carrying
particles can be determined by counting the colonies that has developed. Petri-dishes containing sterile
nutritient agar (NA) medium and potato dextrose agar (PDA) are chosen for the study in this procedure.
The medium chosen is determined by the type of microorganisms to be counted. Nutrient agar
(HiMedia) was used to enumerate airborne pathogenic bacteria, and potato dextrose agar (HiMedia)
was used to obtain the opportunistic fungus from air.The plates are labelled with information such as
the location and time of sample, the length of exposure, and so on. The plates are then left exposed in
the desired location for the specified amount of time, i.e., 30 minutes. All petri dish containing solid
culture media is covered until sampling, at which point the cover is removed and the agar surfaces are
exposed to air for 30 minutes outdoor at 10 am in the morning and 5 pm in the evening. After that, the
petri dishes were sealed and incubated. The nutrient agar plates were kept inverted inside the
bacteriological incubator at 37°C for 24 hours for obtaining aerobic bacterial counts and 3 days at 22 ˚C
for saprophytic bacteria. Molds were cultured on potato dextrose agar; all the exposed plates were
closed and kept straight at a temperature of 25°C for 5–7 days. On agar media, a specific number of
colonies can be seen developing. Each colony symbolises a microorganism-carrying particle that has
settled on the agar surface.The ideal exposure time of 30 minutes was considerable sufficient for
obtaining easily countable number of well isolated colonies. It usually depends on how dusty the air
being sampled is and hence the plate exposed in IPHH hospital was kept for a period of 60 minutes. It's
best to keep the plates about 1 metre above the ground during sampling. The colonies on each plate are
counted and recorded after incubation as the number of bacteria carrying particles settling on a specific
area in a given period of time.
2.1.2 Microscopic Methods
This method allowed air to pass through a membrane filter of 0.45 micron pores size in the direction of
the airflow for trapping the microorganisms, which was then examined microscopically for counting the
number of cells. Acridine orange staining and observation under a fluorescence microscope was done.
The total number of microorganisms in 1 m3 of air is the end outcome. This approach has the benefit of
being able to identify both live and dead microorganisms in the air, as well as those that do not thrive in
culture media.As a result, the number of microorganisms detected is typically one order of magnitude
more than in culture methods. Other biological agents, such as plant pollen, allergic mites, and abiotic
organic dust, can also be detected and5identified. However, the methods have a fundamental flaw:
where the actual determination kind of the species of microorganism is not possible.
Culture- dependent Methods
Microbes are transferred from the air onto the surface of the suitable selective medium such as
mannitol salt agar- MSA (HiMedia) and milk agar- MA (HiMedia). The forms of colonies were counted
after a 24-hour incubation period at the ideal temperature of 37°C, and the result is presented as
CFU/m3 of air. Because a colony can form from a single cell as well as a cluster of cells, the air could
contain more bacteria than the colony forming unit (CFU)as the test suggests. Furthermore, the
procedure enables for the detection of only live cells that are able to proliferate in the selective medium
used. Microbes that have been moved to the culture medium must be revived because they have been
exposed to hazardous conditions.For bacterial identification, Gram's staining was done, and lactophenol
cotton blue staining was done for the fungal identification. Pure cultures were obtained by sub-culturing
distinct colonies on freshly prepared nutrient medium and streaking them sequentially to obtain pure
cultures, while pure fungal cultures were obtained by spot inoculation. Colonial features, microscopy,
and biochemical assays were used to characterise and identify pure bacteria cultures. In bacteriology,
the Gram stain is the most extensively used staining process to distinguish between Gram-positive and
Gram-negative bacteria, also known as a differential staining. This staining technique was used along
with the biochemical tests for identifying the bacterial species based on differences in biochemical
activities among bacterial species. Colony morphology and lactophenol cotton blue (LCB) staining was
used to identify the fungal colonies.
62.2 Statistical Calculation
The results obtained would recorded as the number of colonies obtained (colony forming unit per
meter cube- CFU/m3). The statistical significance of the test results were drawn by calculating the
Arithmetic Mean (AM) and Standard Deviation (SD). The Student’s “one sample t- Test” was used as a
statistical tool tocompares the sample mean with a hypothesized population means for drawing a
significance at p <0.05. All the statistical calculation would be done using Microsoft Excel 2003 and SPSS
version 26 software.
3.1 Identification of microorganisms
The colony counts of bacteria and fungi obtained after a plate exposure time of 30 minutes would be
expressed as CFU/ m3. At least 7 different types of bacterial species and 6 different types of fungi
species were detected from the air during the study. The suspected colonies of bacteria identified
through the colony morphological study and biochemical tests . The fungal colony was detected by
staining and microscopic analysis .In indoor air, the commonest generally would be of fungi found
Aspergillus, Penicilliumand the commonest genera of bacteria found wereStaphylococci, Bacillus and
Pseudomonas. Microbes found in air over populated land areas below altitude of 500ft in clear weather
include spores of Bacillus and ascospores of yeasts, fragments of mycelium and spores of molds, etc. In
outdoor air, common genera of fungi such asCladosporium, and the bacterial genera such as Bacillus,
Micrococcus, Streptococcus, Staphylococcus and Pseudomonas were predominant.6
2 Enumeration and statistical calculations
The air microbial loads of seven different sites of IPHH were determined by taking indoor air samples
(figure1). The results of the study considering the number of colonies, concentration range, arithmetic
mean, standard deviation of bacterial and fungi present in the investigated sites of IPHH are represented
in the table 3, table 4 and 5.The results indicate that the highest bacterial count in CFU/m3air of indoor
samples recorded at 5 pm in Classroom at 30 min exposure, which is 8920 CFU/m3, while the lowest
bacterial CFU/m3air were recorded at
10 a.m. in Hospital ICU at 30min exposure, which is 15CFU/m3 The highest fungal content (CFU/m3) in
air of indoor sample has been recorded at 5 pm in the library at 30 min exposure time, which is
189CFU/m3, while the lowest fungal CFU/m3 air were recorded at 10 am in hospital ICU at 60 min
exposure time, which is 08 CFU/m3
Conclusion
7An assessment of bacteria and fungi in the indoor and outdoor environment were experimentally
investigated. It is understood from the present environmental study that the air quality is better in early
morning as less anthropogenic activity occurs during morning. The road transport affects the air quality.
The road transport systems deteriorate air quality. Seems transportation occurs throughout the day,
thus air quality is affected. Microbial populations in the air may be seasonal or all-year in character,
although their occurrences are generally most intense in summer and autumn. Also in the daytime the
sunlight reduces the number of 77pathogens from air. There is essential to monitor air quality in both
indoor and outdoor environments for microorganisms, in particular with regard to their effects on
human health. Increased awareness of the threats posed by biological air pollution will enable greater
care of the conditions in which people live every day. In particular, the proper ventilation of buildings,
ventilations of all classrooms library and laboratory and prevention of excessive air humidity are all
essential. 7
-Microorganisms in the outdoor air: sources, transit, and variables determining their existence would
also be discussed .
