CHAPTER ONE

INTRODUCTION

1:1. BACKGROUND OF STUDY

Abattoir is a place designed statutorily and designated by approving bodies for the slaughtering
of animals, processing, preserving and storing of meat and by products of livestock for human
consumption. (Alonge, 1991). They are also commonly called slaughter house and are places
where livestock especially animals such as cattle, sheep, pigs, goats e.t.c. are slaughtered or
butchered for food. It can also be known as any assigned place used for the butchering of animals
primarily as meat for man’s consumption. It does not include any place that is situated on a
farmland (Abattoir acts 1998). Slaughtering animals for a household or community dates back as
civilization, public Abattoirs dates back as far as Roman civilization.

The Abattoir is a facility where meat is processed for the supply of protein. A number of
operations take place at a typical Abattoir in Nigeria raging from the temporary storage of the
animal and to washing, cutting, roasting of the skin e.t.c. The processing of meat in most
Abattoirs is crude and constitutes a health and environmental hazard, both in the area of hygiene
of the operation and the disposal of the waste generated. These wastes include solid (bone, dung
e.t.c.) liquid (blood, semen e.t.c.) and gaseous materials. In as much as they are needed for meat
production and processing and also for by products such as hides and skin, they are key sources
of environmental pollution of water, air, and land as a result of processes like bleeding, dressing,
hide removal, removal of internal organs, cuttings and boning e.t.c. this is because they contain
high amounts of liquid wastes, organic solids, and fats and generates large quantities of solid
wastes and wastewaters with biological oxygen demand (BOD) and offensive odour (Osibanjo et
al 2007). Effluents such as blood, hair, fat, bones etc. are peculiar to the Abattoir and according
to NNEP 2000, are potential disease carriers and as such regarded as waste from industries, due
to the complex nature of abattoir wastewater, they can be a source of danger to the environment.

The abattoir operation impacts directly and indirectly on human health in so many ways the
wrongful processing technique which contaminates the meat directly like in the deposition on
carcinogenic compounds on the roasted skin which is consumed in Nigeria directly causing
ingestion of harmful compounds and when waste washes into soil and water, that will also
indirectly impact on human health.

Pollution in Abattoirs is majorly due to the poor hygiene and not adhering to good hygiene and
manufacturing practice. (Akinro et al., 2009). The waste generated are not properly disposed and
are allowed to litter around the Abattoirs and with time are decomposed and washed away into
open drainages, rivers and streams and introduce pathogens into surface waters along surplus
nutrients and also leech into underlying aquifers and contaminate wells that are manually dug.
(Adeyemo et al., 2009).

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1:2. Environmental Pollution

This is the release of any biological, chemical, physical or radiological substances into the
ecosystem (air, water, land or soil) at a rate faster than its threshold and which threatens the life
or survival of man, plants and animals or any biotic system of the environment. In other words,
environmental pollution is any undesirable change in the biogeochemical processes of the earth
that may pose potential danger on the health and survival of humans and other living organisms.
The alteration in the physical, chemical or biological characteristics of the air, water or land
(soil) and that will harmfully affect the survival or activities of man and other flora and fauna or
any abiotic system can be viewed as environmental pollution.

One of our Era’s greatest scourges is air pollution, on account not only if its impact on public
and individual health due to increasing morbidity and mortality. There are many pollutants that
are major factors in disease in humans. Among them Particulate matter (PM), Particles of
Variable but very small diameter, penetrate the respiratory system via inhalation, causing
respiratory and cardiovascular nervous system dysfunctioning and cancer. Despite the fact ozone
in the stratosphere plays a protective role against ultraviolet radiation, it is harmful when in high
concentration at ground level, also affecting the respiratory and cardiovascular system.
Furthermore, nitrogen oxide, sulfur dioxide, volatile organic compounds (VOCs), dioxins, and
polycyclic aromatic hydrocarbons (PAHs) are all considered air pollutants that are harmful to
humans. Carbon monoxide can even provoke direct poisoning when breathed in at high levels.
Heavy metals such as Lead, when absorbed into the human body, can lead to direct poisoning or
chronic obstructive pulmonary Disease (COPD), asthma, bronchiolitis, and also lung cancer,
cardiovascular events, central nervous system dysfunctions and cutaneous disease. Climate
change resulting from environmental pollution affects the geographical distribution of many
infectious diseases, as do natural disasters.

Human activities have an adverse effect on the environment by polluting the water we drink, the
air we breathe, and the soil in which plants grow. Although the industrial revolution was a
success in terms of technology, society, and the provision of multiple services it is also
introduced the production of huge quantities of pollutants emitted into the air that are harmful to
human health. Without any doubt, the global environmental pollution is considered an
international public issue with multiple facets. Clearly urbanization and industrialization are
reaching unprecedented and upsetting proportions worldwide on our era. Anthropogenic air
pollution is one of the biggest public health hazards worldwide given that it accounts for about
Nine Million deaths per year (WHO, 2019).

1:3. BIOAEROSOLS

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Bioaerosols are very small airborne particles (ranging from 0.001 to 100-um) that originate
biologically from plants/animals and can contain living organisms (Georgakopoulos et al, 2009).
Therefore, pathogenic and non-pathogenic dead or alive microorganisms (e.g viruses, bacteria
and fungal) may exist in bioaerosols (Mandal and Brandl 2011). Bioaerosols are easily shifted
from one environment to another because of their small size and light weight (Van Leuken et al.,
2016). In recent years, exposure to Bioaerosols in both occupational and residential
environments has drawn much attention in light of their probable impacts on human health.
Sources of Bioaerosols exposure in occupational activities are diverse enough to include waste
sorting and composting, agricultural and food processing activities, the livestock industry e.t.c
(Pearson et al., 2015). Indeed, the prevalence of diverse respiratory diseases or symptoms
(allergic asthma, rhinitis, airway inflammation etc.) has been reported from workers susceptible
to such exposure. (Beck et al., 2012, Rohr et al., 2015). Bioaerosols were estimated to be
responsible for approximately 5 to 34% of indoor particulate air pollution (Mandal and Brandl,
2011).

Furthermore bioaerosols also contain microorganisms and their components such as fungal,
bacteria, endotoxin, mycotoxins and allergens, such organisms are well known normal
components of both indoor/outdoor air.

1.4 STATEMENT OF PROBLEM

The poor waste management is responsible for the environmental and health hazards associated
with Abattoirs. The hazards have indirectly threatened or endangered the health of residents and
the environment in general. This is because animal waste such as blood, bones, intestinal content,
tissues, hides and skin are scattered in huge piles around the abattoirs. On entering most abattoirs
in Nigeria one immediately sees the glaring evidence of a failed and a broken system. This
includes dilapidated slaughtering and processing facilities, inadequately clean water supplies, no
refrigerators and lack of facilities for the collection and storage of waste. Proper sewage or waste
disposal systems are also lacking. The waste water emanating from Abattoirs pollutes surface
and underground water as well as the air. The pungent stench forces neighboring residents to
shut their windows and doors thus disallowing cross ventilation in homes.

The flaring of these gasses, due to roasting of meat releases a lot of pollutants into the abattoir
environment.

The piled up waste also causes air pollution which subsequently produces methane gas that
intensifies the greenhouse effect on global warming. This study investigates the bioaerosols in an
abattoir community using Swale Market as a case study.

1.5 JUSTIFICATION OF STUDY

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Air pollution of grave consequences

The year 2012 witnessed an estimated 3.7 million premature death out of 7 million that were
attributable to ambient air pollution. Eighty eight (88%) of these deaths were recorded by low
and middle income countries (WHO, 2014) and 4.3 million deaths from household air pollution
(WHO, 2015). The adverse health effects occasioned by particulate matter cannot be over
emphasized. The chemical composition of particulate matter is a vital signature to human healthy
and wealthy living. However, there are scanty reports of the air quality and assessment of
bioaerosols in an abattoir community and thus this study.

1.6. OBJECTIVES OF THE STUDY

The general objective of this study is to examine and assess the air quality and bioaerosols in
four locations around Swale abattoir market.

The specific objectives of this study are

To assess the microbiological quality of the bioaerosols around the Swale abattoir
To know the gases present in the selected locations around the abattoir
To measure the level of particulate matter (PM2.5 and PM10) within the abattoir
environment
To quantify the heavy metals present in the air around the abattoir

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 CHAPTER TWO

LITERATURE REVIEW

2.1 AIR QUALITY

Air quality can be said to be the degree to which the surrounding air is clean. When the threshold
to which the surrounding air is within the acceptable set standards, the index of quality with
respect to such an environment is said to be of high quality and thus not within the acceptable set
standards, the air in that environment by any chemical, physical or biological agent that modifies
the natural characteristics of the atmosphere (Ojukwu and Somerville, 2020).

Urban air quality is a dynamic process involving the continuous emission from stationary and
mobile combustion sources. Mobile sources contribute to the emission of major pollutants over a
given location. These labile pollutants at various level include; gasses (CO 2, NO2, N2O5, SO2,
SO3 etc), photo-chemical oxidant (O3), ozone precursors (hydrocarbons), volatile organic
compounds, heavy metals and black carbon (Costa, 2004).

2.2. PARTICULATE MATTER

Studies have shown a relationship between particulate matter (PM) and adverse health effects,
focusing on either short-term (acute) or long-term (chronic) PM exposure.

Particulate matter (PM) is usually formed in the atmosphere as a result of chemical reactions
between the different pollutants. The penetration of particles is closely dependent on their size
(Wilson et al., 1997). Particulate Matter (PM) was defined as a term for particles by the United
States Environmental Protection Agency (USEPA, 2018). Particulate matter (PM) pollution
includes particles with diameters of 10 micrometers (μm) or smaller, called PM10, and extremely
fine particles with diameters that are generally 2.5 micrometers (μm) and smaller.

Particulate matter contains tiny liquid or solid droplets that can be inhaled and cause serious
health effects (Cheung et al., 2011). Particles <10 μm in diameter (PM10) after inhalation can
invade the lungs and even reach the bloodstream. Fine particles, PM2.5, pose a greater risk to
health (Zhang et al., 2019).

Multiple epidemiological studies have been performed on the health effects of PM. A positive
relation was shown between both short-term and long-term exposures of PM2.5 and acute
nasopharyngitis. In addition, long-term exposure to PM for years was found to be related to
cardiovascular diseases and infant mortality. (Zhang et al., 2019).

Those studies depend on PM2.5 monitors and are restricted in terms of study area or city area
due to a lack of spatially resolved daily PM2.5 concentration data and, in this way, are not
representative of the entire population. Following a recent epidemiological study by the
Department of Environmental Health at Harvard School of Public Health (Kloog et al., 2013), it

5
was reported that, as PM2.5 concentrations vary spatially, an exposure error (Berkson error)
seems to be produced, and the relative magnitudes of the short- and long-term effects are not yet
completely elucidated. The team developed a PM2.5 exposure model based on remote sensing
data for assessing short- and long-term human exposures. This model permits spatial resolution
in short-term effects plus the assessment of long-term effects in the whole population.

Moreover, respiratory diseases and affection of the immune system are registered as long-term
chronic effects. It is worth noting that people with asthma, pneumonia, diabetes, and respiratory
and cardiovascular diseases are especially susceptible and vulnerable to the effects of PM.
PM2.5, followed by PM10, are strongly associated with diverse respiratory system diseases
(Kappos et al., 2004), as their size permits them to pierce interior spaces (Boschi 2012). The
particles produce toxic effects according to their chemical and physical properties. The
components of PM10 and PM2.5 can be organic (polycyclic aromatic hydrocarbons, dioxins,
benzene, 1-3 butadiene) or inorganic (carbon, chlorides, nitrates, sulfates, metals) in nature
(Chueng et al.2011).

Particulate Matter (PM) is divided into four main categories according to type and size (Kumar
et al., 2012). Gas contaminants include PM in aerial masses. Particulate contaminants include
contaminants such as smog, soot, tobacco smoke, oil smoke, fly ash, and cement dust. Biological
Contaminants are microorganisms (bacteria, viruses, fungal, mold, and bacterial spores), cat
allergens, house dust and allergens, and pollen. Types of dust include suspended atmospheric
dust, settling dust, and heavy dust.

Finally, another fact is that the half-lives of PM10 and PM2.5 particles in the atmosphere is
extended due to their tiny dimensions; this permits their long-lasting suspension in the
atmosphere and even their transfer and spread to distant destinations where people and the
environment may be exposed to the same magnitude of pollution (USEPA, 2018). They are able
to change the nutrient balance in watery ecosystems, damage forests and crops, and acidify water
bodies. As stated, PM2.5, due to their tiny size, are causing more serious health effects. These
aforementioned fine particles are the main cause of the “haze” formation in different
metropolitan areas (Kan et al., 2012).

2.3. Polycyclic Aromatic Hydrocarbons (PAHs)

The distribution of PAHs is ubiquitous in the environment, as the atmosphere is the most
important means of their dispersal. They are found in coal and in tar sediments. Moreover, they
are generated through incomplete combustion of organic matter as in the cases of forest fires,
incineration, and engines. PAH compounds, such as benzopyrene, acenaphthylene, anthracene,

6
and fluoranthene are recognized as toxic, mutagenic, and carcinogenic substances. They are an
important risk factor for lung cancer (Abdel-Shafy et al., 2016).

Polycyclic aromatic hydrocarbons (PAHs) are widespread across the globe mainly due to long-
term anthropogenic sources of pollution. The inherent properties of PAHs such as heterocyclic
aromatic ring structures, hydrophobicity, and thermo stability have made them recalcitrant and
highly persistent in the environment. PAH pollutants have been determined to be highly toxic,
mutagenic, carcinogenic, teratogenic, and immunotoxicogenic to various life forms. Therefore,
this review discusses the primary sources of PAH emissions, exposure routes, and toxic effects
on humans, in particular. This review briefly summarizes the physical and chemical PAH
remediation approaches such as membrane filtration, soil washing, adsorption, electrokinetic,
thermal, oxidation, and photocatalytic treatments.

Rapid industrialization and urbanization have resulted in numerous anthropogenic activities,

which dump various pollutants in the environment, including polycyclic aromatic hydrocarbons
(PAHs) (Mojiri et al., 2019). Due to their inherent properties, PAHs are persistent pollutants
having a wide range of biological toxicity; remediation of PAHs from the environment has been
a global concern. The PAH pollutants are ubiquitous, found equally in aquatic and terrestrial
ecosystems as well as in the atmosphere (Adeniji et al., 2019). The PAH pollution, either directly
or indirectly, is strongly affecting the health and well-being of humans, along with other
organisms across the planet (García-Sánchez et al., 2018). The choice of appropriate strategies
for PAH remediation is always critical, as it is highly dependent on two major parameters:
polluted matrix and environmental conditions (Kuppusamy et al., 2017). Different remediation
methods involving physical, chemical, biological, and lately developed integrated approaches
have been continuously applied at varying degree of success. Among the many remediation
approaches, methods based on microorganisms for ecological restoration of PAH-polluted
environments have been a well-evaluated approach (Kuppusamy et al., 2017; Malla et al., 2018;
Mehetre et al., 2019). Recently, the integrated PAH remediation methods have also been
reported for efficient mitigation of PAH pollutants. The aim of this review is to discuss current
knowledge and recent developments in PAH remediation strategies, associated factors, and their
effectiveness as well as limitations.

2.4 Volatile Organic Compounds (VOCs)

Volatile organic compounds (VOCs), such as toluene, benzene, ethylbenzene, and xylene
(Kumar et al., 2014), have been found to be associated with cancer in humans (Molhave et al.,
2014). The use of new products and materials has actually resulted in increased concentrations of
VOCs. VOCs pollute indoor air (Kumar et al., 2014) and may have adverse effects on human
health (Molhave et al., 2014). Short-term and long-term adverse effects on human health are
observed. VOCs are responsible for indoor air smells. Short-term exposure is found to cause
irritation of eyes, nose, throat, and mucosal membranes, while those of long duration exposure
include toxic reactions (Gibb, 2019). Predictable assessment of the toxic effects of complex

7
VOC mixtures is difficult to estimate, as these pollutants can have synergic, antagonistic, or
indifferent effects (Ebersville et al., 2012).

VOCs are responsible for the odor of scents and perfumes as well as pollutants. VOCs play an
important role in communication between animals and plants, e.g. attractants for pollinators,
(Moores 2009) protection from predation, (Karl et al., 2009) and even inter-plant interactions.
(Marlon et al., 2019) Some VOCs are dangerous to human health or cause harm to the
environment. Anthropogenic VOCs are regulated by law, especially indoors, where
concentrations are the highest. Most VOCs are not acutely toxic, but may have long-term chronic
health effects. Most VOCs in earth's atmosphere are biogenic, largely emitted by plants. (WHO,
2019)

Biogenic volatile organic compounds (BVOCs) encompass VOCs emitted by plants, animals, or
microorganisms, and while extremely diverse, are most commonly terpenoids, alcohols, and
carbonyls (methane and carbon monoxide are generally not considered). (Kassomenos et al.,
2012) Not counting methane, biological sources emit an estimated 760 teragrams of carbon per
year in the form of VOCs. (Dherani et al., 2008. The majority of VOCs are produced by plants,
the main compound being isoprene. Small amounts of VOCs are produced by animals and
microbes. (Dockery, 1993) Many VOCs are considered secondary metabolites, which often help
organisms in defense, such as plant defense against herbivore. The strong odor emitted by many
plants consists of green leaf volatiles, a subset of VOCs. Although intended for nearby organisms
to detect and respond to, these volatiles can be detected and communicated through wireless
electronic transmission, by embedding nanosensors and infrared transmitters into the plant
materials themselves.

Emissions are affected by a variety of factors, such as temperature, which determines rates of
volatilization and growth, and sunlight, which determines rates of biosynthesis. Emission occurs
almost exclusively from the leaves, the stomata in particular. VOCs emitted by terrestrial forests
are often oxidized by hydroxyl radicals in the atmosphere; in the absence of NOx pollutants,
VOC photochemistry recycles hydroxyl radicals to create a sustainable biosphere-atmosphere
balance. (Newlands, 2015) Due to recent climate change developments, such as warming and
greater UV radiation, BVOC emissions from plants are generally predicted to increase, thus
upsetting the biosphere-atmosphere interaction and damaging major ecosystems.(NEPIS, 2017)
A major class of VOCs is the terpene class of compounds, such as myrcene (NRC, 2019).
Providing a sense of scale, a forest 62,000 km2 in area, the size of the US state of Pennsylvania,
is estimated to emit 3,400,000 kilograms of terpenes on a typical August day during the growing
season. Researchers investigating mechanisms of induction of genes producing volatile organic
compounds, and the subsequent increase in volatile terpenes, has been achieved in maize using
(Z)-3-hexen-1-ol and other plant hormones. (Spiegel et al., 2019)

The major sources of man-made VOCs are: Fossil fuel use and production, e.g. incompletely
combusted fossil fuels or unintended evaporation of fuels. The most prevalent VOC is ethane, a

8
relatively inert compound (Gibson, 2009). Solvents used in coatings, paints, and inks.
Approximately 12 billion litres of paints are produced annually. Typical solvents are aliphatic
hydrocarbons, ethyl acetate, glycol ethers, and acetone. Motivated by cost, environmental
concerns, and regulation, the paint and coating industries are increasingly shifting toward
aqueous solvents (Kaun et al., 2017). Compressed aerosol products, mainly butane and propane,
estimated to contribute 1.3 billion tonnes of VOC emissions per year globally (Bezitzoglou et al.,
1996). Biomass combustion, especially from rain forests. Although in principle combustion gives
carbon dioxide and water, incomplete combustion affords a variety of VOCs.

2.5. NITROGEN OXIDES (NOX)

NOX gases are usually produced from the reaction among nitrogen and oxygen during
combustion of fuels, such as hydrocarbons, in air; especially at high temperatures, such as in car
engines. (WHO, 2019). In areas of high motor vehicle traffic, such as in large cities, the nitrogen
oxides emitted can be a significant source of air pollution. NO X gases are also produced naturally
by lightning.

The term NOX is chemistry shorthand for molecules containing one nitrogen and one or more
oxygen atom. It is generally meant to include nitrous oxide (N 2O) (WHO, 2019). Although
nitrous oxide is a fairly inert oxide of nitrogen that has many uses as an oxidizer for rockets and
car engines, an anesthetic, and a propellant for aerosol sprays and whipped cream. Nitrous oxide
hardly plays any role in air pollution, although it may have a significant impact on the ozone
layer, (Marlon et al., 2019) and is a significant greenhouse gas.

When NO x and volatile organic compounds (VOCs) react in the presence of sunlight, they form
photochemical smog, a significant form of air pollution. The presence of photochemical smog
increases during the summer when the incident solar radiation is higher. The emitted
hydrocarbons from industrial activities and transportation react with NO X quickly and increase
the concentration of ozone and peroxide compounds, especially peroxyacetyl nitrate (PAN).
(Hashim et al., 2014)

Children, people with lung diseases such as asthma, and people who work or exercise outside are
particularly susceptible to adverse effects of smog such as damage to lung tissue and reduction in
lung function (GUO et al., 2017).

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2.5.1 SOURCES OF NITROGEN OXIDE

2.5.1.1 Natural sources

Nitric oxide is produced during thunderstorms due to the extreme heating and cooling within a
lightning strike. This causes stable molecules such as N 2 and O2 to convert into significant
amounts of NO similar to the process that occurs during high temperature fuel combustion.
(HOU et al., 2008). NOx from lightning can become oxidized to produce nitric acid (HNO 3),
this can be precipitated out as acid rain or deposited onto particles in the air. Elevated production
of NOx from lightning depends on the season and geographic location. The occurrence of
lightning is more common over land near the equator in the inter-tropical convergence zone
(ITCZ) during summer months. (Kan et al., 2012) This area migrates slightly as seasons change.
NOx production from lightning can be observed through satellite observations.

A recent discovery indicated that cosmic ray and solar flares can significantly influence the
number of lightning strikes occurring on Earth. Therefore, space weather can be a major driving
force of lightning-produced atmospheric NO X (Burroughs 2017).

2.5.1.2 Biogenic Sources

Agricultural fertilization and the use of nitrogen fixing plants also contribute to atmospheric NO
X, by promoting nitrogen fixation by microorganisms (Saud, et al., 2012). The nitrification

process transforms ammonia into nitrate. Denitrification is basically the reverse process of
nitrification. During denitrification, nitrate is reduced to nitrite, then NO, then N 2O and finally
nitrogen. Through these processes, NOX is emitted to the atmosphere. (Singh, et al., 2013)

A recent study conducted by the University of California Davis found that adding nitrogen
fertilizer to soil in California is contributing 25 percent or more to state-wide NO X pollution
levels. (Dherani et al., 2008) When nitrogen fertilizer is added to the soil, excess ammonium and
nitrate not used by plants can be converted to NO by microorganisms in the soil, which escapes
into the air. NOX is a precursor for smog formation which is already a known issue for the state
of California. In addition to contributing to smog, when nitrogen fertilizer is added to the soil and
the excess is released in the form of NO X, or leached as nitrate this can be a costly process for the
farming industry.

A 2018 study by the Indiana University determined that forests in the eastern United States can
expect to see increases in NOX and in turn, changes in the types of trees which predominate. Due
to human activity and climate change, the maples, sassafras, and tulip poplar have been pushing

10
out the beneficial oak, beech, and hickory. The team determined that the first three tree species,
maples, sassafras, and tulip poplar, are associated with ammonia-oxidizing bacteria known to
"emit reactive nitrogen from soil." By contrast, the second three tree species, oak, beech and
hickory, are associated with microbes that "absorb reactive nitrogen oxides," and thus can have a
positive impact on the nitrogen oxide component of air quality. Nitrogen oxide release from
forest soils is expected to be highest in Indiana, Illinois, Michigan, Kentucky and Ohio.
(Kassomenos et al., 2012)

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 CHAPTER THREE

MATERIALS AND METHOD

3:1 DESCRIPTION OF SAMPLING AREA

Samples were collected from Swale abattoir community in Yenagoa, Yenagoa local government
area of Bayelsa state. Four sampling locations within Swale community were designated as
sampling point. The first point is the main abattoir where the animal slaughter and dressing is
carried out. The second point is the area where provisions are sold. The third point is the abattoir
road and the last location is the main park at the entrance of the market.

Table 3.1: Map coordinate of sampling locations

LOCATIONS DESCRIPTION LATITUDE LONGITUDE

(NORTH) ( EAST)
LOCATION 1 Swale main abattoir N4054’51.86 E6016’0.42456

LOCATION 2 Swale abattoir Provision market N4054’57.2796 E6016’5.898

LOCATION 3 Swale abattoir road N4055’1.47612 E6015’55.32588

LOCATION 4 Swale market park N4055’6.024 E6015’59.442

3.2 SAMPLE COLLECTION

Samples for this study was collected from the middle of January to the middle of February, 2022.

Sixteen already prepared plates of sterile media consisting of eight nutrient agar plates and eight
Potato Dextrose Agar plates were labeled accordingly and used for sampling each week. Two
plates of nutrient agar and two plates of PDA were exposed for a duration of ten minutes
(10minutes) at each sampling location. The plates were then covered, sealed, labeled
approximately and transported immediately to the lab for incubation. Plates of bacterial growth
were incubated in inverted position in the incubator for 24hours (36 ± 2 0C), while plates for
fungal growth were placed at disinfected designated laboratory table (25± 20c) for 48hours.

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3.3 IDENTIFICATION OF MICROBES

This is a standard feature, concerned with the characterization and speciation of microorganisms,
understanding the type of microorganisms and where it may have come from, forms an important
part of contamination assessment. Identification is also important for maintenance of stock
cultures. Methods used in microbial identification

3.3.1 MORPHOLOGICAL CHARACTERISTICS

Distinct feature of each colony of microbial growth were observed for their specific and
descriptive features. Such features include colony shape, colour, form, edges, elevation e.t.c. this
was well documented.

3.3.2 IDENTIFICATION OF BACTERIAL ISOLATES

Biochemical tests are tests that identify the bacteria on the basis of the presence of certain
enzymes and differences in the biochemical activities of different bacteria were carried out. They
are among the most important methods for microbial identification. The analytical techniques
used to carry out microbial identification have, therefore, become essential to a number of
applications. A range of platforms have been developed to carry out microbial identification.

3.3.2.1 GRAMS STAINING TECHNIQUE

PRINCIPLE

When the bacteria is stained with primary stain Crystal Violet and fixed by the mordant, some of
the bacteria are able to retain the primary stain and some are decolorized by alcohol. The cell
walls of gram positive bacteria have a thick layer of protein-sugar complexes called
peptidoglycan and lipid content is low. Decolorizing the cell causes this thick cell wall to
dehydrate and shrink, which closes the pores in the cell wall and prevents the stain from exiting
the cell. So the ethanol cannot remove the Crystal Violet-Iodine complex that is bound to the
thick layer of peptidoglycan of gram positive bacteria and appears blue or purple in colour.

In case of gram negative bacteria, cell wall also takes up the CV-Iodine complex but due to the
thin layer of peptidoglycan and thick outer layer which is formed of lipids, CV-Iodine complex
gets washed off. When they are exposed to alcohol, decolorizer dissolves the lipids in the cell
walls, which allows the crystal violet-iodine complex to leach out of the cells. Then when again
stained with safranin, they take the stain and appears red in color.

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PROCEDURE

With the aid of a sterile wire loop, a smear of the test organisms was made on a clean grease free
slide

Crystal Violet was poured and kept for about 30 seconds to 1 minutes and rinsed with water.

The slide was then flooded with grams iodine for 1minute and washed under running water

Furthermore, the slide was washed with 95% alcohol or acetone for about 10-20 seconds and
rinsed with water.

Safranin was added for about 1 minute and then washed

The slide was finally air dried, and observed under the microscope

3.3.2.2 CATALASE TEST

This test is used to detect the ability of a bacterial to produce enzyme catalase, an enzyme that
catalyses the release of oxygen from hydrogen peroxide (H 2O2). It is found in most aerobic and
facultative anaerobic bacteria. The main exception is Streptococcus spp. It is not found in
anaerobes. Catalase test is primarily used to differentiate Staphylococcus and Streptococus.

PROCEDURE

A smear of the test bacterial was prepared on a clean slide

A drop of 3% hydrogen peroxide was added unto the slide

The evolution of oxygen bubbles was observed as a positive result while the lack of this is a
negative result.

3.3.2.3 OXIDASE TEST

Oxidase test is helpful in the identification of microorganism having ability to produce

cytochrome oxidase enzyme. The test helps to differentiate oxidase positive Pseudomononacea
and negative Enterobacteriacea families. The enzyme oxidase catalyzes the oxidation of reduced
cytochrome by molecular oxygen. Kovac’s oxidase reagent (that contains tetramethyl-p-
phenylenediamine dihydrochloride) is the main reagent used in the oxidase test. The dye serves
as an alternate substrate for the cytochrome oxidase reaction. In the reduced state, the reagent is
colorless but when oxidize, it becomes purple.

PROCEDURE

A smear of the sample was made on the filter paper

And few drops of 1 % kovac’s was added on the smear

14
 After 20seconds a colour change was observed

3.3.2.4 METHYL RED TEST

The Methyl red (MR) test detects the production of sufficient acid during the fermentation of
glucose and the maintenance of conditions such that the pH of an old culture is sustained below a
value of about 4.5, as shown by a change in the colour of the Methyl red indicator which is
added at the end of the period of incubation.

PROCEDURE

Methyl red broth was inoculated with a pure culture of the organism

After inoculation it was incubated at 35o-37o C for 48hours

5 drops of methyl red reagent per 5ml of broth was added

The broth medium was observed for colour change

3.3.2.5 CITRATE TEST

Citrate agar is used to test an organism’s ability to utilize citrate as a source of energy. The
medium contains citrate as the sole carbon source and inorganic ammonium salts as the sole
source of nitrogen. Bacteria that can grow on this medium produce an enzyme, citrate-permease,
capable of converting citrate to pyruvate can then enter the organism’s metabolic cycle for the
production of energy. Growth is indicative of utilization of citrate, an intermediate metabolite in
Krebs cycle. When the bacteria metabolize citrate, the ammonium salts are broken to ammonia,
which increases alkalinity. The shift in pH turns the bromthymol blue indicator in the medium
from green to blue above pH 7.6.

PROCEDURE

A slant was prepared using Simmon’s Citrate Agar

The slant was streaked back and forth with a light inoculum picked from the isolated colony

The slant was incubated aerobically for 35-370 C for 4-7days

It was observed for a color change

3.3.2.6 THE TRIPLE SUGAR IRON TEST

The Triple Sugar Iron (TSI) test is a microbiological test named for its ability to test a
microorganism’s ability to ferment sugars and to produce hydrogen sulfide.

15
PROCEDURE

A well isolated colony was touched with a straight inoculation needle

TSI was inoculated by stabbing through the center of the medium to the bottom of the tube and
then the surface of the agar slant was streaked

The tube was capped was loosely and incubated at 350-370C in ambient air for 18 to 24hours

medium was examined for reaction

3.3.2.7 UREASE TEST

Urea is the product of decarboxylation of amino acids. Hydrolysis of urea produces ammonia
and CO2. The formation of ammonia alkalinizes the medium, and the pH shift is detected by the
color change of phenol red from light orange at pH 6.8 to magenta (pink) at pH 8.1. Rapid
urease-positive organisms turn the entire medium pink within 24 hours.

PROCEDURE

Surface of the urea agar slant was streaked with a portion of a well-isolated colony and
inoculated with 1 to 2 drops from an overnight brain-heart infusion broth culture. The tube was
capped loosely and incubated at 350-370C in ambient air for 48hours

Tube was examined for development of pink color for as long as 7days.

3.3.3 AIR SAMPLING ANALYSIS

Air sampling is a process used to determine what airborne contaminants are present in an
environment. It uses special instruments to detect contaminants such as gases, vapors, dusts and
fibers in the air. The equipment used is the mini volume potable air sampler.

The mini volume potable ampler is used to collect total suspended particulate matter in ambient
air. The sample filter traps the total suspended particulate such as PM 2.5 and PM 10 particles as
the air passes through the instrument.

PROCEDURE

The sampler was carefully transported to the field site and installed in the mounting cradle. It is
positioned to air intake upward in an unobstructed area at least 30 cm from any obstacle to air
flow.

The sampler was placed on a firm level surface.

The clean preseparator/filter holder assembly was removed from the plastic transport bag and the
protective plastic bag was removed from the rain cap.

16
The following information was recorded on the PM Field Data Sheet: number of the filter, the
battery ID, sampler ID, ambient temperature and pressure, flow meter reading, and elapsed time
meter reading.

The (ON/AUTO/OFF) button was turned on to obtain the beginning flow rate in order to start the
pump. Using the Flow Rate Adjustment control the flow meter was set to flow within
specifications for the project temperature and pressure conditions. The reading of the flow meter
was taken from the center of the ball.

The ON/AUTO/OFF button was pressed twice to stop the pump.

The ON/AUTO/OFF button was pressed to set the timer to "Auto" mode. be in “.

The pump and timer assembly was pressed back into the sampler body and the snap buttons was
secured.

Using the hoisting pole, the bale assembly bar was hooked and the sampler was raised as
vertically as possible, to the mounting cradle. This position not only more easily accommodates
the sampler's weight, but prevents the hook from hitting and possibly dislodging or breaking the
preseparator/filter holder assembly.

Fig 3.1: The researcher collecting samples for analyses

17
3.3.3.1 ASSESING FOR PARTICULATE MATTER

An aerosol particle counter works on the principal of either light scattering or light blocking. An
aerosol stream is drawn through a chamber with a light source (either laser based light or white
light). When a particle is illuminated by this light beam, it is redirected or absorbed. Light
scattered by a single particle in a specific direction in relation to the original direction has a
unique signature which relates to the size of the particle. This allows for sizing and counting of
individual particles.

PROCEDURE

The power button was turned on and the particle counters was set and allow to read for
10minutes.

18
 CHAPTER FOUR

Presented in Figure 4.1 is the total heterotrophic bacterial count for the four locations across a
four week duration.

500

450

400

350

300
Week 1
250 Week 2
200 Week 3
Week 4
150

100

50

0
Swale main abattoir Swale provision Swale abattoir road Swale market park
market

Fig. 4.1: Total heterotrophic bacteria count at Swale main abattoir

From the figure above, it can be observed that samples obtained at Week 1 had the highest
bacterial load count at all locations except Location 4 (Swale market park). This unconnected
with the fact that the main abattoir is where the highest anthropogenic activities such as
slaughtering of animals, dressing, roasting (by flaring different organic and inorganic fuel
sources) and discharge of animal gut wastes. These activities however, may increase microbial
load around this location.

At Week 2, the Swale provision market area recorded the highest bacteria load (145cfu) while
the least count was at the main abattoir (88cfu). Nevertheless the overall microbial load was
lower than Week 1 for all the sampling locations.

At the third week of sampling, even though the bacteria load was generally lower than the
previous weeks, the highest load was recorded at Swale main park (39cfu) while the least count
was at the abattoir road (10cfu).

At Week 4, there was a slight increase in the general microbial count. Yet, the count was at the
Swale main park (138cfu) and least at Swale provision market with (51cfu). Worthy of note is
19
 the fact that microbial load was on a consistent decrease till Week 3. This may be due to the
onset of the rains as the weeks progressed. Nevertheless, the fourth week had a slight increase
in bacterial load across all sampling locations.

Table 4.2 Identification of bacterial isolates

Sample OX CAT M VP CIT TSI UREA GS MOT PROBABLE

R

ISO 1 + + - - + - +/- - + Pseudomonads flourescene

ISO 2 + + - - + - + + + Micrococcus luteus

ISO 3 - + - - + +/- - - - Acinetobacter spp

ISO 4 - + + - +/- - + - Corynebacter spp

ISO 5 - + + - - + - - + Eschericha coli

ISO 6 + + + + + + - - + Aeromonas hydrophila

ISO 7 - - + - +/- - - + - Clostridium spp

ISO 8 - + + + + - + + - Staphylococcus aureus

ISO 9 - + - +/- + +/- + - - Klebsiella pneumonia

ISO 10 + + - - + - - - + Pseudomonas aeruginosa

KEYS:

ISO: ISOLATE

OX: OXIDASE

CAT: CATALASE

MR: METHYL RED

VP: VOGES PROSKAUER

CIT: CITRATE

TSI: TRIPLE SUGAR IRON

GS: GRAM STAININ

20
The aforementioned were the biochemical tests that were carried out to enhance the
identification of the bacterial isolates. Table 4.2 shows the biochemical reaction in identifying
the probable organisms present in this study. The organisms isolated from the sampled
bioaerosols were tentatively identified.

100
90
80
70
60
50
40
30
20 WEEK 1
10 WEEK 2
0 WEEK 3
s s pp pp li ila pp s ia sa
cen teu r s r s co p h s reu on no
WEEK 4
e s lu e e ia o m u m i
r us ct ct ich r iu sa u ug
flou cc ba ba er h yd trid cu ne a er
o o o h s c p
as oc et et Es as Cl
o co la na
s
on icr A cin ryn on y lo siel o
om M o om h eb
C
er ap Kl om
eud A St e ud
Ps Ps

Fig. 4.2: Distribution of bacteria isolates at Swale main abattoir

The distribution of bacteria isolate at Swale main abattoir is presented in Figure 4.2 bacteria
isolates all had their highest number in the first week of sampling at this location. The bacteria
with the highest count was Pseudomonasflourescens, while the least was Pseudomonas
aureginosa. Week 1 recorded the highest number of bacteria isolates and Week 3 recorded the
least number of isolate count.

21
 13% 14%

2%
3%
8% Pseudomonas flourescens
Micrococcus luteus
Acinetobacter spp
Corynebacter spp
Escherichia coli
11% Aeromonas hydrophila
Clostridium spp
27% Staphylococcus aureus
Klebsiella pneumonia
Pseudomonas aeruginosa
12%

6% 4%

Fig. 4.3: Percentage occurrence of bacterial isolates at Swale main abattoir

From Figure 4.3, Clostridium spp had the highest percentage of occurrence at 27%, followed by
Pseudomonads fluorescens at 14%. From the distribution presented in Figure 4.2 it can be
observed that the occurrence of Pseudomonas flourescens reduced at Week 2, 3, and 4. While
Clostridium spp. had a slight increase on Week 2 and 4.

22
 60

50

40

30

20 Week 1
Week 2
10 Week 3
Week 4
0
ns s p p li la p s a sa
ce teu sp sp co hi sp r eu oni no
es lu er er ia op m u m gi
r ct ct ich dr iu a
ou us ba ba
s ne
u ru
fl occ to e her
s hy s trid ccu p s ae
s oc e yn c a co lla a
ad cin or Es on Cl
o
on
on icr A C m hylo bsie m
m M ro ap e o
do St Kl ud
u Ae e
Ps
e Ps

Fig 4.4 Distribution of bacteria isolates at Swale provision market

From the distribution of bacteria isolate presented in Figure 4.4, it can be observed that week one
had the highest number of occurrence compared to week two and week four which had more
isolates, although the difference is minimal. Week one had six isolates, week two and week four
had seven isolates while week three had the least isolate which is five.

23
 9%
3% 20%
Pseudomonas flourescens
9%
Micrococcus luteus
Acinetobacter spp
Corynebacter spp
Escherichia coli
13% Aeromonas hydrophila
Clostridium spp
21% Staphylococcus aureus
Klebsiella pneumonia
10% Pseudomonas aeruginosa

4% 5%
6%

Fig. 4.5: Percentage occurrence of bacterial isolates for Swale provision market

From the presentation of Figure 4.6 it can be observed that Clostridium spp had the highest
percentage of occurrence at 21%, followed closely by Pseudomonas flourescene at 20%, while
klebsiella pneumonia had the least percentage at 3%. From Figure 4.4 it was observed that
Clostridium spp and Pseudomonas flourecene occur at almost the same percentage, this may be
due to the fact that Clostridium spp and Pseudomonas flourecene grow well when there is
minimal water activity of 0.90 (Jay, 2005).

24
 50
45
40
35
30
25
20
15 Week 1
10 Week 2
5 Week 3
0 Week 4
ns us p p oli ila p us nia sa
sce lute r sp r sp ia c ph m sp ure o ino
re s cte cte ch ro iu a um ug
flou ccu oba eba eri hyd trid ccus pne aer
o et h
as oc yn Esc nas Clos loco ella nas
on icr Acin Cor o y si o
d om M r om aph Kleb dom
Ae t
eu S eu
Ps Ps

Fig. 4.6: Distribution of bacterial isolates at Swale abattoir road

The distribution of identified bacteria isolated at Swale abattoir road is presented in Figure 4.6.
Week 1 had the highest occurrence of organisms and fewer isolates were observed in Week 2,
while Week 3 had the least number of isolates. Furthermore, Week 4 recorded higher number of
isolates with fluctuating numbers. This changes may be due to the onset of the rainy season was
as at Week 4 of sampling.

25
 7%

11% 24%
Pseudomonas flourescens
Micrococcus luteus
6% Acinetobacter spp
Corynebacter spp
Escherichia coli
Aeromonas hydrophila
8%
Clostridium spp
12% Staphylococcus aureus
Klebsiella pneumonia
5% Pseudomonas aeruginosa
5%
3% 17%

Fig. 4.7: Percentage occurrence of bacterial isolates Swale abattoir road

From the presentation of Figure 4.7 it can observed that Pseudomonas flourescene had the
highest percentage of occurrence at 24%, followed closely by Acinetobacter spp. While
Corynetobacter spp had the lowest percentage occurrence 3%. The low percentage of
corynebacter spp. may be due to the fact that corynebacter spp. grow more on broth media than
agar media.

26
 50
45
40
35
30
25
20
15 Week 1
10 Week 2
Week 3
5
Week 4
0
ns s p p li ila p us ia sa
ce teu r sp r sp a co ph sp re on no
es lu e e i o m u m gi
ou
r us ct ct ich yd
r iu sa ne
u
er
u
fl occ oba eba h er h tr id ccu p a
as oc et yn Es
c as os co la as
on icr cin Cor on Cl y lo siel on
A om h eb om
om M
er ap Kl
eud A St eud
Ps Ps

Fig.4.8. Distribution of bacterial isolates at Swale market park

From the presentation of Figure 4.8 it can be observed that week four had a spike in bacterial
count despite the distance from the main abattoir were the highest anthropogenic activities are
carried out, week two had the highest number of occurring isolate, while week four had least
number of occurrence.

27
 8%
15%
7%

6% Pseudomonas flourescens
Micrococcus luteus
Acinetobacter spp
15% Corynebacter spp
Escherichia coli
Aeromonas hydrophila
16% Clostridium spp
Staphylococcus aureus
Klebsiella pneumonia
Pseudomonas aeruginosa
5% 17%

8%
4%

Fig. 4.9: Percentage occurrence of bacterial isolates at Swale market park

From the presentation of Figure 4.9 it can be observed that Pseudomonas flourescens,
Micrococcus luteus, Acinetobacter spp. Clostridium spp. had the highest range of percentage of
occurrence while Corynebacter spp. Escherichia coli, Aeromonas hydrophila, Staphylococcus
aureus, Klebsiella pneumonia, and Pseudomonas aeruginosa has the lowest range of percentage
occurrence.

28
 40

35

30

25
Week 1
20 Week 2
15 Week 3
Week 4
10

0
Swale main abat- Swale provision Swale abattoir Swale market park
toir market road

Fig.4.10. GENERAL FUNGAL COUNT

Presentation from Figure 4.10 it can be observed that all locations had recorded the a high fungal
load count for week 1 and all were reducing at a descending order. This is as a result of the
distance from where the main anthropogenic activities is being carried out, this shows that the
distance from this area reduces the fungal load count.

At Week 2, the Swale market park area recorded the highest fungal load (35cfu) while the least
count was at the abattoir road (7cfu).

At the third week of sampling, even though the bacteria load was generally lower than the
previous weeks, the highest load was recorded at Swale market park (14cfu) while the least count
was at the Swale main abattoir and the abattoir road (2cfu) respectively.

At Week 4, there was a decrease in the general microbial count and the highest recorded count
was at the Swale provision market (5cfu). The Swale main abattoir and Swale abattoir road had
the least count (1cfu) respectively. Worthy of note is the fact that microbial load had a constant
decreasing order from the Week 1 to Week 4.

29
TABLE 4.3: FUNGAL IDENTIFICATION

ISO MORPHOLOGY ORGANISMS

ISO 1 Dark green filamentous Trichoderma viride

ISO 2 White fluffy filamentous Coccidioides immities

ISO 3 White filamentous with yellowish colour at Fusarium oxysporium

center

ISO4 Dark brown filamentous Aspergillus niger

ISO 5 Dark green filamentous with yellowish Pennicillium chrysogenum

colour at center

ISO6 Dark brown and folded tops Cladosporium cladosporiodes

ISO 7 Dark green with white round edges Aspergillus fumigatuss

KEYS:

ISO: ISOLATE

30
 9
8
7
6
5
4
3
Week 1
2
Week 2
1 Week 3
0 Week 4
de es m r m s us
iri iti riu n ige nu iode
gat
v m o e i
a m p s g or um
erm esi x ys gillu r yso
osp sf
d d o er ch d lu
o io m p cla il
ich cid riu As uim m erg
Tr Co
c sa ll u p
Fu ici ri As
e nn spo
P o
ad
Cl

Fig. 4.11 Distribution of fungal isolates at Swale main abattoir

From the distribution of fungal isolates presented in Figure 4.11 the occurrence of each fungal is
observed and Week1 has the highest fungal count followed closely by Week 2, Week 3 and
Week 4 has lowest occurrence of fungal.

31
 19%

Trichoderma viride
4%
Coccidioides immities
49% Fusarium oxysporium
6% Aspergillus niger
Pennicilium chrysogenum
Cladosporium cladosporiodes
10% Aspergillus fumigatus

8%
4%

Fig. 4.12. Percentage of occurrence of fungal isolates at Swale main abattoir

From presentation of figure 4.12 it shows Trichoderma viride having the highest percentage of
occurrence of (49%), Coccidiodes immities and Cladosporium cladosporiodes the lowest
percentage at (4%) each.

32
 10
8
6
4
2 Week1
Week 2
0 Week 3
de es m
ige
r m es us Week 4
viri m
iti
oriu n enu r iod igat
a m p s og o m
erm esi x ys gillu r ys osp s fu
id o r ch d
od io m sp
e
cla illu
r ich cid r iu A iu m m e rg
T c sa cil iu p
Co Fu nni por As
Pe ad
o
Cl

Fig. 4.13. Distribution of fungal isolates at Swale provision market

From the distribution of fungal isolates presented in Figure 4.13 Week1 had the highest number
of occurrence, followed closely by week 2, while Week3 and Week 4 had the least number of
occurrence.

33
 24%
29%
Trichoderma viride
Coccidiodes immities
Fusarium oxysporium
Aspergillus niger
11% Pennicilium chrysogenum
3% Cladosporium cladosporiodes
Aspergillus fumigatus
10% 17%
6%

Fig 4.14: Percentage occurrence of fungal isolates at Swale provision market

From the presentation of figure 4.14 the percentage of occurrence of fungal in location 2(abattoir
road) it shows Aspergillus fumigatus with a low percentage rate at Week 1, but at Week 2, it
increases and starts decreasing from Week3 to Week 4. Where Trichoderane viride has the
highest percentage of occurrence at Week 4 followed by Clasdosporium clasporiodes.

34
 8
6
4
2
Week 1
0 Week 2
de es m r u s s Week 3
iri iti r iu nige sog iode
gatu
v m po ry i Week 4
a m s or m
erm esi x ys gillu ch
osp s fu
id o er im d lu
od io m sp lu cla gil
ir ch ic d r iu A icil m er
T c sa nn riu sp
Co Fu Pe o A
p
os
lad
C

Fig. 4.15: Distribution of fungal isolate at Swale abattoir road

From the presentation of Figure 4.15 it can be observed that these location ( Swale provision
market) had few isolates occurrence, week one had five isolates, week two had three isolates, and
week three had two isolates, week four had the least isolate just one isolate was seen on week
four.

35
 11% 14%

13% Trichoderma viride

Coccidioides immities
Fusarium oxysporium
4% Aspergillus niger
Pennicillium chrysogenum
Cladosporium cladosporiodes
38% Aspergillus fumigatus

20%

Fig 4.16: Percentage occurrence of fungal isolates at Swale abattoir road

From the presentation of figure 4.16 it shows the percentage occurrence of fungal isolates on
location 3(Swale provision market). It shows increase of the percentage of occurrence of
Coccidiodes immities (38%) followed byAspergillus niger at 20%.

36
 8
7
6
5
4
3
Week 1
2 Week 2
1 Week 3
Week 4
0
de es m r s
iv ri iti r iu ige n um ode
m n e i
a m po s g or
erm esi x ys gillu r yso
osp
od id o er ch d
ir ch ic d
io ir u
m
A sp m cla
T c sa lliu riu
m
Co Fu nici o
n p
Pe os
lad
C

Fig 4.17: Distribution of fungal isolates at Swale market park

From the distribution of fungal isolates presented at Figure 4.17 it was observed that there was a
spike in the occurrence of the isolate for each week except week four, these may be due to the
distance from the main abattoir to the bridge.

37
 9%

30%

Trichoderma viride
Coccidioides immities
29%
Fusarium oxysporium
Aspergillus niger
Pennicillium chrysogenum
Cladosporium cladosporiodes
Aspergillus fumigatus

5% 19%
5%
4%

Fig.4.18: Percentage occurrence of fungal isolate at Swale Market Park

From the presentation of figure 4.18 it shows the percentage of occurrence of fungal isolates for
location four (Swale market park) in this figure, Trichoderma viride had the highestpercentage at
and (30%) followed by Cladosporium cladosporiodes at 29%

From the Previous Figures it was observed that the Percentage of occurrence rate for each
identified organisms were fluctuating and decreasing in a descending order this may be as a
result of the level of Contamination at the sampling area. Findings from other research works
( Ahmad et al., 2005) it was deduced that Aspergillus fumigatus have great potential in reducing
heavy metals concentration in contaminated environment, likewise Fusarium oxysporium which
was identified to be effective in reducing the concentration of Cu and Pb (Chandrakar et al,
2012). Also Trichoderma viride was identified to have a high accumulating potential for Ni, Cr
(Joshi et al, 2011). These findings brought out the fact that the occurrence of these/ some of this
organisms are not just harmful to humans and the environment but that they are also beneficial as
stated above.

38
TABLE 4.3: CONSTITUENTS OF AIR SAMPLE

S/N Station Ar CU Pb Fe Ni Cr Cd PAHS TPH

(mg/kg)
1 <0.001 <0.021 0.086 0.036 0.033 0.028 0.061 14.508 32.10
3 8

Temperature
Wind speed
humidity

direction
Relative
Station

PM 2.5

TVOC
PM 10

Sound
Wind
COX

NOX

SOX
S/N

(ppm)
1 Swale 0.04 0.02 0.04 1.75 2.2 1.75 626 58.4 E/W 0.08 79.7
Main
Abattoir
Key:

Ar: Argon

Cu: Cupper

Pb: Lead

PAH: Polycyclic Aromatic Hydrocarbons

TPH: Total Petroleum Hydrocarbon

COx: Carbon monoxide

Nox: Nitrogen oxide

SOx: Sulfur oxide

PM: Particulate matter

TEMP: Temperature

TVOC: Total Volatile Organic Compound

39
Analysis of these constituents of air shown in Table 4.3 shows the level of concentrations of each
heavy metals and also the particulate matter. Lead (Pb) is a naturally occurring element found in
small amounts in the earth crust. While it has some beneficial uses it can be toxic to humans and
animals causing health effects. Exposure to lead comes from human activities example burning
and use of fossil fuels. Lead can affect almost every organ and system in the body. Results from
this study shows the level of concentration of lead to be 0.086mg/kg, comparing these result to
that of the National Ambient Quality Standard which is 0.02 it shows the result exceeded the
standard limit which is harmful and unhealthy to those around the area. Nickel (Ni) it is a
chemical element that is highly ductile corrosion and oxidation resistant and recyclable. These
characteristics make it essential for building infrastructure communication energy supply etc.
nickel is found in ambient air at very low levels as a result of releases from oil coal combustion
and wood incineration. Exposure to high level of nickel by inhalation cause severe damage to the
lungs and kidney. (ATSDR 1997). Comparing the level of concentration of Nickel which is
0.0333 to that of the National Ambient Quality standard which 0.02 it shows that it exceeds the
standard limit, though the difference is slight it is also dangerous.Chromium (Cr) is a trace
element critical to human health and wellbeing. It is a significant environmental threat.
Excessive exposure could lead to higher levels of accumulation in human and animal tissues
leading to toxins and detrimental health effects. Such as coughing, itching and burning sensation,
sores in the nose which leads to nose bleeding. Chromium from this result when compared with
the National ambient quality standard exceeded the standard limit of 0.02 to its
0.08mg/kgCadmium (Cd) is a toxic non-essential transition metal that poses a health risk for
both humans and animals. It occurs naturally in the environment as a pollutant that is derived
from agricultural and industrial uses example burning of fossil fuels such as coal. Acute
exposure of high level of Cadmium in human leads to lung infections such as bronchial and
pulmonary irritation. Cadmium shows a level of concentration on the result as 0.061 which when
compared to the standard limit of 0.02 is high and this is dangerous to the environment and
people around.Polycyclic Aromatic Hydrocarbon (PAH) are class of chemicals that occur
naturally in coal, crude oil and gasoline. They are produced when coal, oil, gas, wood are burned
and in this case roasting in abattoir. PAH generated from these sources can bind to or form small
particles in air. Epidemiological studies shows that PAH are associated with reduced lung
function and cardiovascular diseases. The continuous burning and roasting shows its effect on
the environment when PAH was compared to the standard limit of 0.2, result from this study
showed that the level of concentration PAH surpass the limit at a great length, PAH level in this
study is 14.508mg/kg, the difference on these is extremely and dangerous to the environment and
people living on it. This is as a result of the everyday activity in the abattoir.

Carbon monoxide. It’s a colorless, odorless gas produced by burning gasoline wood e.t.c it is
known to cause Nausea, blurred vision and other effects when exposed to it at a high level. The
level of carbon monoxide in this result is 0.04mg/kg compared to the standard limit it shows that
its level of concentration did not exceed the limit. Nitrogen oxide are a mixture of gasses that are
composed of nitrogen and oxygen, it is one of the main ingredients involved in the formation of

40
ground level ozone which can trigger serious respiratory problems, it also reacts to form nitrate
particles and acidic aerosols contributing to the formation of acid rain. The result shows the level
of concentration of Nitrogen oxide 0.02mg/kg compared to the standard limit of 0.053mg/kg it
does not exceed the standard limit. Sulfur oxide (Sox) it comprises of both gaseous and
particulate chemical species, it is an important air pollutant that acidifies ecosystems and forms
harmful fine particulate matter on the atmosphere. It causes respiratory illness by making
breathing more difficult. The level of concentration of Sulfur oxide from the result is 0.04mg/kg,
compared to the standard limit it did not exceed it. Particulate matter (PM 2.5) is an air pollutant
that is a concern for people’s health when levels in air are high, they are able to travel deeply into
the respiratory tract, reaching the lungs. Exposure to this particle can cause short term health
effects such as eye, nose, throat and lung irritation, coughing, sneezing e.t.c it can also affect
lung function and worsen medical conditions such as asthma and heart disease. Pm primarily
come from burning of fuels such as wood, heating oil or coal and natural sources such as forest
and grass fires. The result of Pm 2.5 and Pm 10 which is 1.75ppm and 6.26ppm respectively
shows a high level of exceedances to the standard limit of 0.012ppm and 0.035ppm of the
National ambient quality standard, from this result, the level of contamination in the abattoir area
and the effect of this activities on the environment and to the people living in the area is observed
and this calls for immediate attention by health officials.

41
 CHAPTER FIVE

CONCLUSION

The presence of bioaerosols has been suspected as the cause of various human diseases not only
infectious/respiratory but also cancer. Stating that the bioaerosols of the abattoir environment
indicated the occurrence of the following pathogenic organisms even though the number
fluctuates over the weeks nevertheless, the microorganisms are always there and can have
access to humans through inhalations or terminal dispositions or even by ingestion, so the
occurrence of these organisms calls for health concerns of both the workers in the abattoir and
even farther communities because these organisms are always present in the air, and the air is
always moving, it can go to environments farther than the immediate place where it is being
generated, so it calls for general health concern. It is expected to have better abattoir practices
whereby activities like roasting should be minimized, open fire roasting should be limited or
eradicated and then good manufacturing practices should be introduced such that animal waste
from animal guts are nut dumped indiscrimately anywhere without treatment, because all these
contributes to the microbial load in the bioaerosols of the abattoir environment. Furthermore, the
air also have constituents that pose to be health hazards to humans such as the Lead, Nickel,
Cadmium, Chromium, as well as the Polycyclic aromatic hydrocarbons, Total petroleum
hydrocarbons, Total volatile organic compounds. These constituents should not be present in the
environment where humans inhabit. The presence of these microbial and heavy metals in the air
is a significant pollutant in the environment, their presence have shown that the study indicate
pollution in the air around the abattoir environment.

RECOMMENDATION

From the result of this study, it is recommended that:

 Abattoir operation/facilities should sited far away from residential areas.

 The use of modern facilities should be encouraged, waste should be properly treated
before discharged in the environment
 Open fire roasting of cow skin should be discouraged.
 Periodic inspection of abattoir facilities and procedures should be carried by out by
professionals in related fields to avoid contaminations of the air.
 Government should enact policies that will ensure better abattoir practice for the
processing of animals
 Governments and appropriate authorities should ensure regular supervision of abattoir
activities

42
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 APPENDIX

APPENDIX 1

MATERIALS AND REAGENTS

Media (Nutrient agar and Potato dextrose agar), Distilled water, Cotton wool, Methylated spirit,
Measuring cylinder, Conical flask, Tape, Marker, Aluminum foil, Filter paper, Spatula, Forceps,
Wire loop, Glass slides, Funnel, Steam sterilizer (Autoclave), Incubator, Mini potable air
sampler.

REAGENTS

Hydrogen peroxide, Oxidase disc, Tryptophan, Glucose phosphate broth, Naphtol 5%,Potassium
hydroxide, Phynol red indicator, Crystal Violet, Iodine,Safranin

APPENDIX 2

TYPES OF CULTURE MEDIA USED

The media used for the microbiological assay in this study include Nutrient agar and Potato
dextrose agar (PDA). These media were prepared following manufacturer’s instructions.
Furthermore, 0.5g of Streptomycin was added to PDA per litre to inhibit the bacteria growth
before being poured. The agar were sterilized by autoclaving at 121 0c for 15min and cooled
down to about 450c before pouring aseptically into sterile plates. The plates were swirled to avoid
bubbles and subsequently allowed to set.

NUTRIENT AGAR

Nutrient agar is made with various nutrients which allow the growth of a wide variety of
microorganisms that do not usually require specific nutrients or supplements. The primary
constituents of the media are peptone, beef extract, and agar. In addition to these nutrients, some
vitamins and some trace ingredients necessary for the growth of bacteria are also added. The
peptone is the source of nitrogen or protein that acts as a source of amino acids for bacteria. The
beef extract is the primary source of carbon which is essential for the formation of carbohydrates
in the bacteria. It also contains other components like some vitamins, different trace minerals,
organic compounds, and salts, which further enhance the growth of different organisms. Agar is
the solidifying agent that provides a stable surface for the organism to grow on, which allows for
the observation of colony morphology and enumeration of the organism.

49
Preparation of Nutrient Agar

Work bench was disinfected with methylated spirit using a weighing balance 14grams of
Nutrient agar was measured and to 500ml of distilled water in a conical flask

Conical flask was sealed with tape and aluminum foil

Suspension was heated to boiling to dissolve the medium completely

Dissolved medium is then autoclaved at 1210C for 15mins
Conical flask was taken out after autoclaving and allowed to cool
Media was poured into sterile petri plates under sterile conditions

POTATO DEXTROSE AGAR

Potatoes Dextrose Agar (PDA) is used for the cultivation of fungal. It is a general purpose
medium for yeasts and molds that can be supplemented with acid or antibiotics to inhibit bacteria
growth.

Principle

Potato Dextrose Agar is composed of dehydrated potato infusion and Dextrose that encourages
luxuriant fungal growth. Agar is added as the solidifying agent. Many standard procedures use a
specified amount of sterile tartaric acid (10%) to lower the pH of this medium to 3.5+/-0.1,
inhibiting bacteria growth. Chloramphenicol acts as a selective agent to inhibit bacterial
overgrowth of competing microorganisms from mixed specimens, while permitting the selective
isolation of fungal.

Preparation of PDA

A weighing balance was used to measure 19.5grams of Potato dextrose agar into 500ml of
distilled water

Conical flask was sealed with tape and aluminium foil

Suspension was heated to boiling point to dissolve the medium completely

Dissolved medium was autoclaved at 1210C for 15mins

Conical flask was taken out after autoclaving and allowed to cool
0.5gram of streptomycin was added to the prepared medium
Media was poured into sterile petri plates under sterile conditions

50
 APPENDIX 3

PURIFICATION OF ISOLATES

Bacteria Subculture Procedure

Work bench was disinfected

Bunsen burner was turned on

Each colony to be sub cultured was marked

Wire loop was flamed and allowed to cool

Wire loop was used to scoop a little amount of the colony into the fresh plates

Wire loop was used to streak the media surface

Plate was labeled according to its colony description

Fungal Subculture Procedure

Work bench was disinfected

Bunsen burner was turned on

Each fungal colony was marked

Forcep was cleaned with methylated spirit and used to pick the colony and transferred to a fresh
prepared PDA plates

Forcep was cleaned after collection of each colony.

SLANT PREPARATION PROCEDURE

Double strength of nutrient agar was prepared 4ml of the prepared nutrient agar was dispensed
into the Bejoule bottles and sterilized.

Bottles was placed in a slant position and allowed to solidify

51
 APPENDIX 4

HETEROTROPHIC BACTERIAL COUNT

LOCATIONS WEEK 1 WEEK 2 WEEK 3 WEEK 4

Swale Main Abattoir 429 88 20 49

Swale Provision Market Section 204 145 34 51

Abattoir Road 128 94 10 55

Swale Bridge 82 92 39 138

APPENDIX 5

Percentage occurrence of bacterial isolates at Swale main abattoir

S/N ISOLATES Wk1 Wk1 WK WK2 Wk3 WK3 Wk4 WK4

% 2 % % %

1 Pseudomonas flourescens 88 20.5 - - 4 20 8 16.2

2 Micrococcus luteus 79 18.4 - - - - 6 12.2

3 Acinetobacter spp 58 13.5 26 29.5 - - - -

4 Corynebacter spp 70 16.3 - - 3 15 9 18.4

5 Escherichia coli 69 16.1 - - - - - -

6 Aeromonas hydrophila 65 15.1 - - 2 10 - -

7 Clostridium spp - - 48 54.5 5 25 14 28.6

8 Staphylococcus aureus - - 7 8.1 - - 2 4.1

9 Klebsiella pneumonia - - 7 8.1 - - - -

10 Pseudomonas aeruginosa - - - - 6 30 10 20

52
 APPENDIX 6

Percentage Occurrence of Bacterial Isolate At Swale Provision Market

S/ ISOLATES WK WK1 WK WK2 WK WK3 WK WK4

N 1 % 2 % 3 % 4 %

1 Pseudomonas flourescens 28 13.7 - - 15 44.1 11 21.5

2 Micrococcus luteus 54 26.5 31 21.4 - - 46 5.9

3 Acinetobacter spp 26 12.7 12 8.3 6 17.6 28 -

4 Corynebacter spp - - 6 4.1 2 5.9 6 11.8

5 Escherichia coli 22 10.7 - - 4 11.8 - -

6 Aeromonas hydrophila - - 10 6.9 - - 4 7.8

7 Clostridium spp - - 44 30.3 5 14.7 20 39.2

8 Staphylococcus aureus 40 19.6 22 15.2 - - - -

9 Klebsiella pneumonia - - - - 2 5.9 4 7.8

10 Pseudomonas aeruginosa 34 16.6 20 13.8 - - 3 5.9

53
 APPENDIX 7

Percentage Occurrence of Bacterial Isolates At Swale Abattoir Road

S/ ISOLATES WK 1 WK1 WK2 WK2 WK WK WK WK4

N % % 3 3% 4 %

1 Pseudomonas flourescens 28 21.9 - - 5 50 18 32.7

2 Micrococcus luteus 24 18.8 14 14.9 - - 11 20

3 Acinetobacter spp 46 35.9 22 23.4 - - 8 14.5

4 Corynebacter spp - - - - - - 2 3.6

5 Escherichia coli 30 23.4 - - - - - -

6 Aeromonas hydrophila - - 3 3.2 2 20 - -

7 Clostridium spp - - 20 21.3 - - 7 12.7

8 Staphylococcus aureus - - 15 15.9 - - 5 9.1

9 Klebsiella pneumonia - - 11 11.7 3 30 4 7.3

10 Pseudomonas aeruginosa - - 9 9.6 2 20 - -

54
 APPENDIX 8

Percentage Occurrence of Bacterial Isolates At Swale Market Park

S/N ISOLATES WK 1 WK WK2 WK2 WK3 WK3 WK4 WK4

1% % % %

1 Pseudomonas flourescens 10 12.1 20 21.7 10 10.9 16 11.6

2 Micrococcus luteus 8 9.8 13 14.1 - - 46 33.3

3 Acinetobacter spp 12 14.6 14 15.2 12 13.0 28 20.3

4 Corynebacter spp 6 7.3 - - 3 3.3 6 4.3

5 Escherichia coli - - - - - - 11 7.9

6 Aeromonas hydrophila - - 4 4.3 - - - -

7 Clostridium spp 23 28.0 16 17.4 8 8.7 10 72

8 Staphylococcus aureus 14 17.1 - - - - 8 5.7

9 Klebsiella pneumonia 9 10.9 7 7.6 - - 9 6.5

10 Pseudomonas aeruginosa - - 18 19.6 6 6.5 4 2.9

APPENDIX 9

Total Heterotrophic Fungal Count

LOATIONS WEEK 1 WEEK 2 WEEK 3 WEEK 4

SWALE MAIN ABATTOIR 35 13 2 1

SWALE PROVISION MARKET 27 14 7 5

ABATTOIR ROAD 26 7 2 1

SWALE BRIDGE 22 15 14 2

55
 APPENDIX 10

Percentage Occurrence of Fungal Isolates at Swale Main Abattoir

S/ ISOLATES WK1 WK1 WK WK2 WK3 WK 4 WK4

N % 2 % % %

1 Trichoderma viride 5 14.2 3 23.1 100 4 57.1

2 Coccidioides immities 3 8.6 1 7.6 - - -

3 Fusarium oxysporium 6 17.1 2 15.3 - - -

4 Aspergillus niger 7 20 1 7.7 - 1 14.2

5 Pennicillium chrysogenum 6 17.1 1 7.7 - - -

6 Cladosporium cladosporiodes - - 2 15.3 - - -

7 Aspergillus fumigatus 8 22.9 3 23.1 - 2 28.6

APPENDIX 11

Percentage Occurrence at Swale Provision Market

S/N ISOLATES Wk1 WK1 WK WK2 WK WK4 WK4

% 2 % 3% %

1 Trichoderma viride 9 33.3 2 14.2 28.5 3 60

2 Coccidioides immities 4 14.8 - - - - -

3 Fusarium oxysporium 5 18.5 3 21.4 - 2 40

4 Aspergillus niger 8 29.6 - - - - -

5 Pennicillium chrysogenum 1 3.7 2 14.2 28.5 - -

6 Cladosporium cladosporiodes 3 11.1 - - - 2 40

7 Aspergillus fumigatus 6 18.5 7 50 42.9 - -

56
 APPENDIX 12

Percentage Occurrence of Fungal Isolates at Swale Abattoir Road

S/N ISOLATES WK WK1 WK2 WK2 WK WK3 WK4 WK4

1 % % 3 % %

1 Trichoderma viride 7 26.9 2 28.7 - - - -

2 Coccidioides immities 1 3.8 - - 1 50 1 100

3 Fusarium oxysporium - - - - - - - -

4 Aspergillus niger 6 23.1 4 57.1 - - - -

5 Pennicillium chrysogenum 4 15.4 - - - - - -

6 Cladosporium - - - - 1 50 - -
cladosporiodes

7 Aspergillus fumigatus 8 30.7 1 14.3 - - - -

APPENDIX 13

Percentage Occurrence of Fungal Isolates at Swale Market Park

S/N ISOLATES WK1 WK1 WK2 WK2 WK3 WK3 WK WK4

% % % 4 %

1 Trichoderma viride 4 18.1 3 100 - - 1 50

2 Coccidioides immities 2 9.1 1 6.7 6 42.8 1 50

3 Fusarium oxysporium - - 2 13.3 1 7.1 - -

4 Aspergillus niger 3 13.6 - - 2 14.2 - -

5 Pennicillium chrysogenum - - 4 26.7 - - - -

6 Cladosporium 7 31.8 3 100 4 28.6 - -

cladosporiodes

7 Aspergillus fumigatus 6 27.3 2 23.1 1 7.21 - -

57