CHAPTER ONE

1.0 INTRODUCTION
1.1 BACKGROUND OF THE STUDY

A meat pie is a savoury pie with a filling of meat and other savoury ingredients. Principally

popular in Europe, Australia, New Zealand and South Africa. Meat pie differ in pastry in the

sense that a pastry is typically a more portable oWn the go items, as opposed to more

convectional pie Meat pie is one of the most popular snacks in Nigeria; the best Nigerian

meat pie is moist, yummy having a filling of minced meat, potatoes and carrot. Vegetable

such as green pepper may be added as a matter of choice, (Musa et al., 2015).

Our society appeared to be a social design characterised by expanded portability expansive

individuals with schedule work time with less family or domestic centred exercises, such as

cooking. This circumstance result in meat pie been taken out and vendors are on the increment

and responsibility for good manufacturing practices of food such as sanitary measures and

proper good handling has been transferred from individual/families to the food vendors who

rarely enforce such practices (Musa et al., 2015).

Foodborne diseases are diseases resulting from ingestion of bacteria, toxins and cells produced

by microorganism present in food. Food borne illness is a major international health

problem with a consequent economic reduction. It has been estimated that several pathogens

are associated with beef product (meat pie) which include Escherichia coli, Salmonella spp,

Listeria monocytogens Campypylobacter jejuni, Clostridium perfrigens, Toxoplasma gondii,

Staphylococcus aureus accessed for approximately 3.3-12 million cases as food borne illness

(Buzby et al., 2017). Important bacterial food borne pathogen may be implicated either of

1
the two primary type of food released diseases: food borne infection and food borne

intoxications.

A food borne infection involves the ingestion of pathogen, followed by growth in the host,

including invasion and or the release of toxin (Prescott et al., 2018). Some of the major diseases

of this type associated with beef product include Salmonellosis, Listeriosis and hemorrhagic

colitis caused by a strain of Escherichia coli known as Enterohemorrhagic Escherichia coli

(Dolobes et al., 2011).

Food intoxication produces symptoms after food is consumed because of the growth of the

disease causing microorganisms (bacteria) is not required (Prescott et al., 2018). Major

diseases of this type associated with meat pie include botulism and staphylococcal food

intoxication. Food supply issues of processed meat is usually as a result of contamination

(bacterial) introduced exogenously during activities such as harvesting, processing and

preparation (Torok et al., 2017) or endogenous contamination due to the presence of

bacterial pathogens inherent in the cattle and such serve as major reservoir and vehicle for

meat of these pathogens.

The student population usually consumes meat pie in large numbers. Issues of

gastrointestinal disturbances are usually associated with consumption of such product when

they are prepared, handled or served in unhygienic ways. Meat pie is often the snack of

choice in most social gathering. When such products are prepared far inadvance of service

and held in warm temperature (temperature abuse) multiplication of some bacterial and

growth of their toxins is encouraged and tends to increase.

2
In most countries the most common food borne illness is Staphylococcus food

intoxication. Enterotoxigenic Staphylococcus strains and E. coli strains have been so latent

from food implicated to illness. E. coli and S. aureus are named flora in humans and animals.

Their presence in food is an indication of excessive human handling. They produce disease

when the bacterial contaminate the food. They produce some enzymes which are very harmful

and extra cellular substances some of which are heat stable enterotoxin that render food

dangerous even though it appears normal (Prescott et al., 2018).

Once the bacteria has produced toxin, the food can be extensively and properly cooked

killing the bacterial without destroying their toxin. Many of the toxin are gene based that is

carried on plasmid. The intensity of the sign and symptoms may vary with amount of

contaminated food ingested and susceptibility of individual to toxins. Escherichia coli is

commonly used as a surrogate indicator. Its presences in food (meat pie) generally indicate

direct or indirect food combination. However in Nigeria, a number of foods has been

reported to have high incidence of bacteria but their limited information on the health

challenges from foodborne diseases from meat pie retailed within a highly populous

community.

1.2 STATEMENT OF PROBLEM

Food borne infections and illness is a major international health problem with consequent

economic reduction. Food contamination is a major problem associated with food and snacks in

our society today. In recent times, food borne illness is becoming an alarming concern involving

a broad range of diseases, caused by many bacterial, viral, parasitic and chemical contaminants.

It is upon this premise that this project is embark upon.

3
1.3 SCOPE OF STUDY

The objective of this study is focused on the bacteriological assessment state of meat pie

sold in some selected eateries in Auchi Edo State, Nigeria, characterising and identifying the

bacterial isolates and to suggest possible ways of prevention and controls of the occurrence

of these pathogenic bacteria in meat pie.

1.4 SIGNIFICANC OF STUDY

This research will provide information on microbial levels in the various meat pie in the study

location, which will help in food quality and safety decision-making. Information on microbial

composition of these meat pies will serve as useful information for consumers.

1.5 AIM AND OBJECTIVES

1.5.1 AIM

The aim of this study is to determine the bacteriological quality assessment of meat pie

sold in selected entries in Auchi.

1.5.2 OBJECTIVES

The specific objective of this study are to:

1. Isolate bacteria from meat pie sold in selected eatries in Auchi

2. Identify isolates using cultural morphology and Grams reaction

3. To characterize isolate using biochemical test.

1.6 DEFINITION OF TERMS

Microbial: Relating to or characteristic of a microorganism, especially a bacterium causing

disease or fermentation

4
Pathogen: As an organism causing disease to its host, with the severity of the disease symptoms

referred to as virulence. Pathogens are taxonomically widely diverse and comprise viruses and

bacteria as well as unicellular and multicellular eukaryotes.

Salmonella: Is a genus of rod-shaped gram-negative bacteria of the family

Enterobacteriaceae

Escherichia coli: It is a gram-negative, facultative anaerobic, rod-shaped, coliform beterium of

the genus Escherichia coli Meat is commonly found in the lower intestine of warm-blooded

organism.

5
i
 CHAPTER TWO
2.0 LITERATURE REVIEW
2.1 MAJOR PATHOGEN ASSOCIATED WITH MEAT PIE

Bacteria can be found in many foods including meat pie, if the food is not prepared properly.

Bacteria in food cause ingestion, especially when numerous. Bacteria did not stop growing

after it have been eaten and might even further grow in intestines causing illness. This is

called food intoxication, which is caused by consumption of toxins produced in food by

bacterial growth. It is not the bacteria itself that causes illness but the toxin, which does not

after the appearance, odour of flavour of the food. Major pathogen associate with meat pie

are salmonella sp, Escherichia coli, Listeria monocytogenes, Campylobacter jejuni,

Closridiumperfrigens, Toxoplamagondii and Staphylococcus aureus. Waiter, (2017).

i. Salmonella

Once eaten the food infected by Salmonella bacteria it causes Salmonellosis. Salmonella

will continue to grow in the intestine after it is ingested, setting up an infection and causing

illness. The severity of the illness depends on the size of the dose, the resistance of the host

and the specific strain of Salmonella. It is spread through indirect and direct contact with

intestinal content or excrement of animals, including humans often by hands that are not

washed after using the toilet.

ii. Escherichia coli

Escherichia coli belong to the family of microorganisms called coliforms. Most times

Escherichia coli are helpful to humans living in the intestine and preventing the growth of

more harmful pathogen. One strain, however, often cause a deadly disease canned beef is most

commonly associated with Escherichia coli but other foods such as raw milk, unpasteurized
6
apple juice and cider, sprout, lettuce and spinach have been also implicated. It is prevented

by thorough washing and cooking of the raw product and by avoiding recontamination of

cooked meat with raw meat.

Cattle present a major reservoir of Escherichia coli 0157:117 (Whipp et al., 2014) and while

present at low frequencies pose significant hazards in terms of human disease. This means that

a significant number of animals will carry these and other highly undesirable human

pathogens into abattoir environment and processes, leading to direct and indirect

contamination of equipment and exposed meat surface during slaughter and dressing

activities (Gill, 2018).

Environmental conditions that affect the growth of bacteria can include storage temperature,

moisture availability and physical state of the meat pie. Spoilage bacteria that find condition

most favourable will begin to grow even at refrigerated temperature.

2.2 HEALTH IMPLICATIONS OF CONSUMPTION OF CONTAMINATED MEAT

PIE
2.2.1 Staphylococcal Food Poisoning/Intoxication

Staphylococcus aureus the causative agent o staphylococcus food poisoning is a gram

positive coccus resistant to drying and radiation, found in nasal passage and on skin of

humans and other mammals worldwide. From these sources, it can readily enter food. If the

bacterial is allowed to incubate in meat pie, it produces heat stable enterotoxins that render

the food dangerous even if it appears normal. Once the bacterial have produced toxin, the

food can be extensively and properly cooked, killing the bacterial vegetative cells without

destroying the toxin (Prescott et al.,2018).

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Thirteen (13) different enterotoxin have been identified; enterotoxin A, B, C1, C2, D and E

are most common (Enterotoxin A and B are super antigen) the intensity of the signs and

symptoms may vary with the amount of contaminated food ingested and susceptibility of the

individual to toxin. Typical symptoms include severe abdominal pain, cramps, diarrhoea,

vomiting and nausea. The onset of the symptoms is rapid (usually 1-8 hours) and of short

duration (usually less than 24 hours). The intensity rate of staphylococcal food poisoning is

negligible among healthy individuals. Diagnosis is based on symptoms or laboratory

identification of bacteria from foods by animal toxicity test or an antibody based methods.

Treatment is with fluid electrolyte replacement prevention and control of personel responsible for

food preparation and distribution (Prescott et al., 2018).

2.2.2 Infection due to Escherichia coli

Escherichia coli commonly abbreviated as E. coli is named after (Theodore Escherich) is

a gram negative, facultatively anaerobe rod shaped, non-spore-forming bacterium. It can live in a

wide range of substrate. It is commonly found in the lower intestine of warm blooded organism

(endotherms). Most E. coli strains are harmless but some such as 0157 :H7 are occasionally

responsible for recalls. Optimal growth of E. coli occurs at 37oC (98.6F) but some laboratory

strains can multiply at temperature of up to 49 o C (120.2F). E. coli is a member of the

Escherichia family, enterobacteriaceae. Contaminated food and water are the major source

by which the bacteria are stored. Food poisoning resulting from E. coli is usually caused by

eating unwashed vegetables or undercooked meat. Selected trains can cause a wide variety

of infection in hospitals and community settings. These include gastroenteritis, urinary

infection and inflammatory dysentery (Splanger, 2012)

8
Certain strain of E. coli such as 0157:H70121 & 0104:H21, produce potentially lethal toxin. E.

coli is notorious for causing serious and even life threatening complications such as

haemolytic-uremic syndrome. E. coli habour both heat stable and heat labile enterotoxin. A

subgroup called enterohaemorrhagic E. coli (EHEC) can cause severe potentially fatal illness

known as haemorrhagic colitis which is characterised by bloody diarrhoea and severe

abdominal pain (Dolobes and Doyle, 2011)

Transmission of pathogenic E. coli often occurs via faecal-oral route. Common causes of

transmission include unhygienic food preparation, farm contamination due to manure,

fertilisation, and irrigation of crops with contaminated grey water or raw sewage, direct

consumption of sewage water. Dairy and beef cattle are primary the reservoir of E. coli 0157:

H7 and they can carry it asymptomatically and shed it in their faeces. Food products

associated with E. coli outbreaks include new ground beef, raw milk, sausages, meat pie and

hamburger. According to the United States Food and Drug Administration, the faecal-oral

cycle of transmission can be disrupted by cooking food properly, preventing cross

contamination involving barriers such as gloves for food workers and including health care

policies.

Salmonellosis

Salmonella is a genus of rod shaped, gram negative, non-spore forming and

predominantly motile enteric bacteria with diameter around 0.7-1 .5ʮm, length from 2-1

5ʮm and flagella which aid in all movement (peritrichous). They cause illness like typhoid fever,

paratyphoid and salmonellosis. The initial source of the bacterium is the intestinal tract of

9
birds and other animals. Human acquire the bacterial from contaminated food such as beef

product (meat pie), egg, egg product or water (Prescott et al., 2018).

Once the bacterium is in the body the incubation time is only about 8-48 hours. The

disease results from a true food-borne infection because the bacterial multiply and invade

the intestinal mucosa, where they multiply and invade intestinal mucosa, where they

produce an enterotoxin and a cytotoxin that destroy the epithelial cell. Abdominal pain,

cramps, diarrhoea, nausea, vomiting, and fever are the most prominent symptoms, which

usually passed for 2-5 days but can last for several weeks. During the acute phase of the

disease, as many as one billion Salmonella can be found per gram of faces. Laboratory

diagnosis is by isolation of the bacterium from food or from patient’s stool. Treatment is with

fluid and electrolyte replacement. Preventions depend on good processing practises, proper

refrigeration and adequate cooking (Prescott et al., 2018).

2.2.2 Infection Due To Clostridium Perfrigens

Clostridium perfrigens formerly known as Clostridium welchii is a gram positive rod shaped

anaerobic spore forming bacterium of the genus Clostridium. Clostridium perfrigens is ever

present in nature and can be found as normal component of decaying vegetation, marine

sediment, the intestinal tract of humans (Prescott et al., 2008).

Infection due to Clostridium perfrigens show evidence of tissue necrosis, bacteraemia,

emphysematous cholecystitis and gas gangrene, which is also known as Clostridial

myonecrosis. In the united states and united kingdom, clostridium perfrigens bacteria are the

third most common cause of food borne illness with poorly prepared meat and poultry as the

main culprit in harbouring the bacterium. The clostridium perfrigens enterotoxin (CPE)

10
mediating the disease is heat labile (dies at 74 oC) and can be detected in contaminated food if

not properly heated. Incubation time is between 6-24 (commonly 10-12) hours after ingestion of

contaminated food. Often, meat is well prepared but too far in advance of human

consumption since Clostridium perfrigens form spores that can stand cooking temperatures is

allowed to stand for long enough germination occurs and infection bacterial colonies develop.

Symptoms typically include abdominal cramp and diarrhoea, vomiting and usual fever. The

whole course usually resolves within 24 hours.

In blood agar plates, Clostridium perfrigens grown anaerobically produces betahaemolytic

flat spreading, rough translucent colonies with irregular margins (Prescott et al., 2018)

2.2.3 Listeriosis

Listeria monocytogenes is a common gram positive rod that can be isolated from soil vegetation

and many animal reservoirs. Recent evidence suggests that a substantial number of cases of

human listerosis are attributed to the food borne transmission of Listeria monocytogenes.

Listeria out breaks have been traced to sources such as contaminated milk, soft cheese,

vegetable and meat. Unlike many of the foodborne pathogens which cause primarily

gastrointestinal illness, listeria monoctogenes cause inclusive syndrome such as meningitis,

sepsis, and still birth (Prescott et at., 2018).

Diagnosis of Listeriosis is by Culture. Treatment is by intravenous administration of either

ampicillin or penicillin because Listeria monoctogenes is frequently isolated from food, the

USDA (US Department of Agriculture) and manufactures are putting measures to reduce

the contamination of food product by this bacterium (Prescott et al., 2018).

11
2.3 GENERAL PREVENTION AND CONTROL OF FOOD MEASURES
HAZARDS

The critical control point (CCP) according to a study published in journal of food safety in

2004 in a point step procedure in a food process at which control can be applied and as a

result, food hazard can be prevented, eliminated or reduced to acceptable level. Food

processors must use food CCP limits that have been scientifically validated to prevent

growth of pathogens (government should provide basic amenities such as steady power

supply and access to portable water. Education of handlers is equally important. Above all

high hygiene should be maintained in all food production sectors (Anon, 2012).

2.4 PREVIOUS STUDIES AND INVESTIGATIONS

Data on issue of food borne diseases are well documented worldwide. Foodborne illness is a

major international health problem with consequent economic reduction. Foods borne disease

are disease resulting from ingestion of bacteria, toxin and cells produced by microorganisms

(Doyle et al., 2019). Outbreaks of food borne diseases are caused by food that are

contaminated intrinsically or that become contaminated during harvesting, processing or

preparation (extrinsically) (Toroket al., 2017).

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 CHAPTER THREE
3.0 MATERIALS AND METHODS
3.1: AREA OF STUDY

This study was conducted at federal polytechnic Auchi, Department of Biological Science

Laboratory and Technology. Auchi is located in the northern part of Edo state within the

coordinates of Latitude 7o04’N and Longitude 6o16’E. It is situated in the south geographical

zone of Nigeria with a population of over 150,000 people according to the 2006 population

census. It is approximately one hundred and thirty kilometer (130km) away from Benin City the

capital of Edo state. Auchi is the Headquarter of Esako west LGA and has witnessed territorial

development from rural-urban migration. It is bounded to the North by Jattu, to the South by

Aviele, to the East by Iyakpi and to the West by Owan local government Area. It is also the seat

of the federal polytechnic, Auchi, Edo State, Nigeria.

3.2: SAMPLE COLLECTION

Meat pie were obtained randomly from different meat pie vendors at popular meat pie spots in

Sabo, Auchi Edo State. The samples was immediately wrapped in sterile aluminum foil to

prevent contamination and then transported to the Laboratory in the Department of Micro

Biology for analysis.

3.3: LIST OF APPARATUS AND EXPERIMENTAL REAGENTS

3.3.1: APPARATUS
The apparatus used include Petri-dish, Test tubes, Beakers, Colony counter, Weighing balance,

Mac-Conkey agar, Nutrient agar, Conical flask, Hand gloves,

Wire loop, Masking tape, Cotton wool, Microscope, Incubator, Autoclave, Syringe and needles,

Microscope slides, Distilled water, Pasteur pipette, Moltar and Pistle.

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3.3.2: REGENTS USED
The following Regents was used: Crystal violet, Lugol iodine, Acetone, Safranin, Hydrogen

peroxide.

3.3.3: STERIZATION OF EQUIPMENT

All materials that was used in the course of this project practical such as glass wares was

properly washed with detergent and water to remove dirty and contamination and dried properly.

This was followed with proper sterilization of the material in an autoclave at a temperature of

1210C for 15 minutes according to Nwachukwu and Oguocha (2004).

3.4: PREPARATION OF AGAR

3.4.1: NUTRIENT AGAR (NA)

28g of nutrient agar powder was weighed using a weighing balane and dispensed into a

beaker 500mls of distilled water was measuring using a measuring cylinder and dispersed into a

beaker containing the agar powder. It was stirred to dissolve for 10mins. The mixture was

transferred into a conical flask and the neck of the flask was curved with Catton wool wrapped

into aluminum foil. It was auto Dave at temperature of 121 0C and pressure 15psi for 15-20 mins,

the sterilized agar was allowed to cool for about 450C and then ascetically poured into petri

dishes and allowed to set.

3.5 PREPARATION OF SAMPLE

One (1g) of meat pie was cut using a sterile razor blade from each of the sample and was

transported separately into a sterile mortar and pestle and was masarrated and then dissolved in

10ml of distilled water.

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3.5.1: SERIAL DILUTIONS
Six (6) test tubes labelled 10-1, 10-2, 10-3, 10-4, 10-5, 10-6, containing 1ml of dissolved meat pie

solution each was used for the serial dilution test. 1ml of the stock solution was introduced into

the first test tube labelled 10-1 and was well homogenized. 1ml of the homogenized solution from

the first test tube was transferred from the first tube into the second test tube labelled 10 -2 and

was well homogenized. 1ml of homogenized solution from the second test tube was transferred

from the second test tube into the third test tube labelled 10 -3 and was well homogenized. 1ml of

homogenized solution from the third test tube was transferred from the third test tube into the

fourth test tube labelled 10-4 and was well homogenized. The process continued until it gets to

the last test tube labelled 10-6 and one (1ml) from the last test tube was discarded.

3.5.2: INOCULATION AND INCUBATION

0.5ml Aliquot from the serial dilution sample was added into six (6) Petri dishes containing

molten agar. It was then allowed to solidify. The plates were incubated invertedly at 37 OC for

24hrs.

3.5.3: BACTERIAL COLONY COUNT

The represented agar plate incubated was visualized under a colony counting machine and was

used to count the total bacterial count (labtech/ India) and result was expressed as colony

forming unit per milliliter (cfu/ml) at the end of the count as reported by Olayemi et al (2011).

3.6 IDENTIFICATION OF BACTERIAL ISOLATES

Identification of the bacterial isolates was performed using classical method based on their

morphological and biochemical characteristics with reference to systematic manual of

bacteriology described by Cheesbrough (2004).

15
3.6.1: IDENTIFICATION OF BACTERIA (GRAM REACTION)
3.6.1.1 GRAM STAINING:
Gram staining reaction has the wide application that is capable of distinguishing virtually all

bacteria into one of two large group, Gram positive or Gram negative as described by Dr. Hans

Christian Gram. (1884). A drop of water was placed on a clean slide and a speck of bacterial

growth was taken from the culture. The speck was emulsified on the slide to make a thin smear.

The smear was then being passed over a flame to heat fix. It was then flooded with crystal vislet

for 1 minutes, washed with distilled water and iodine was added for 1 minutes, washed with

distilled water and decolorized with Acetone was added for few seconds i.e. 2-5 secs and washed

with distilled water. The smear was finally flooded with safrain to counter stain for 45secs. It

was then washed with distilled water and allowed to air dry. Oil immersion was used and viewed

with X100 objective lens. Purple coloration denoted gram positive.

3.6.1.2: CATALASE TEST

This was used to differentiate those bacteria that produce enzyme catalase. Catalase test was

carried out as described by Cheesebrough (2006). A little portion of the bacteria growth was

transferred with a sterilized wire loop on a clean glass slide and 2ml of 3% hydrogen peroxide

(H202) was dropped on the glass slide and observed for the production of gas bubble which

indicates a positive reaction. This test was used to identify Staphylococcus aureus and Bacillus

cereus (James 2001).

(i) MOTILITY TEST

Motility test is aimed at identifying motile bacteria, it was carried out as described by Baiyewu

(2007). Motility test can sometimes be referred to as the way an organism grow on solid media

16
and it is determined by the presence or absence of flagella. Tubes containing the motility

medium were inoculated by making a fane stab with a loopful of the culture to a depth of 1-2cm

and it was then incubated at 37 OC for 24hrs. Motility is observed by spreading of the organism

outwards from the stab area.

(ii) CITRATE TEST

This test is based on the ability of an organism to use citrate as its source of Carbon. With aid of

an inoculating loop, a loopful of the isolates was aseptically streaked on the surface of a citrate

agar and then incubated at 37 OC for 24hrs. A bright blue coloration on the surface of the citrate

agar will be observed.

(iii) OXIDASE TEST

With the aid of an inoculating loop, fresh culture of the isolates was smeared on the surface of

the oxidase strip moistened with deionized water and then observed for color change within 30-

60secs. A purple to deep blue color was observed.

(v) COAGULASE TEST

This was used to identify Staphylococcus aureus which produces the coagulase enzyme which

cause plasma to clot by converting fibrinogen to fibrin. The slide method was used. A drop of

sterile distilled water was placed on each end of a sterile slide. Then a colony of the test

organism was emulsified on each spot to make two thick suspensions. A loopful of plasma was

added to one of the suspensions and mixed gently. The slide was checked for clumping or

clotting of the organisms within 10seconds. Plasma is not added to the second suspension which

serves as control.

17
 CHAPTER FOUR
4.0 RESULTS AND DISCUSSION
4.1: RESULT
The result of the bacteriological analysis of meat pie sold in Auchi is tabulated below. Table 1

present cultural and morphological characteristics of isolates. Table 2 present total viable cell

count of sample isolates. Table 3 present biochemical characteristics of isolates. Table 4 present

percentage occurrence of isolates in samples.

Table 1: cultural and morphological characteristics of the bacterial isolates.

Cultural Morphological Gram reaction Suggestive organism

Characteristics characteristics

Smooth latose fermenter entire Irregular rod like shape - Escherichia coli
discrete mucoit colonies. Size
2-3cm Elevation convex Color
pink

Smooth pale pink colony. Size +

1 micro diameter. Elevation Cocci (round shape) Staphylococcus
raised Structure opaque grape like clusters. aureus

Size 1.5-3mm x 0.5mm. Rod-shape in chains + Bacillus cereus

Elevation low converse irregular colonies

18
Table 2: Total number of variable cell count of sample isolates

S/N Sample Dilution Factor No of Colonies Colony forming

unit cfu/ml

1 A 10-1 45 4.5x10-1

10-3 37 3.7x10-3

10-5 28 2.8x10-8

2 B 10-1 32 3.2x10-1

10-3 25 2.5x10-3

10-5 15 1.5x10-5

3 C 10-1 35 3.5x10-1

10-3 19 1.9x10-3

10-5 12 1.2x10-5

4 D 10-1 30 3.0x10-1

10-3 17 1.7x10-3

10-5 10 1.0x10-5

Key: A = Sabo, B = Poly Road, C= Big taste, D = Matice

4.2 DISCUSSION

Meat products ( meat pie) basically contains all the nutrient necessary for microbial growth and

metabolism, making it susceptible to microbial contamination. In this study, three organisms

were isolated from meat pie sample following inoculation on Mac Conkey agar after incubation

for 24hrs at 37OC. The organism isolated were suggested of Escherichia coli, Staphylococcus

19
aureus, and Bacillus cereus based on cultural and biochemical characteristic following

Bergeys manual of determinative bacteriology.

The result was in consonance with the work of Yah et al (2009) who isolated

Staphylococcus aureus, Escherichia coli, Salmonella spp, Pseudomoneas spp and Bacillus

spp from meat pie sample in Benin city, Edo State. In their study, it was stated that the

possible occurrence of Staphylococcus aureus in sample was because of the certain amount

of salt present in the meat of the meat pie sample that the growth or Staphylococcus aureus

whereas, the presence of some members of enterio-bacteriacea family was due to

contamination form instestinal slaughtered animal.

The result of table 2 shows total number of viable cell count of sample isolate collector

from 4.5x10-3 to 2.8x10-3 to 3.0x10-5 respectively. The high load of isolates on sample

depicts unhygienic condition of meat handling by vendors. The result is in agreement with

the work of Abdulahi, (2014) who stated that high load isolate on meatpie sample arise as a

result of poor handling by meat pie vendors.

20
 CHAPTER FIVE
5.0 CONCLUSION AND RECOMMENDATION
5.1 CONCLUSION

Various bacteria genera were isolated and identified from the samples, These

include, Staphylococcus, Escherichia, and Bacillus. The result revealed that the samples

(meat pie) sold in these eateries in Auchi were directly or indirectly contaminated with

high levels of pathogenic bacteria, most especially Staphylococcus spp and Escherichia

spp. This study clearly confirmed the deplorable state of ready to eat food (meat pie).

However occurrence of the pathogens can be essentially reduced or prevented by

employing good manufacturing practises (GMP).

5.2 RECOMMENDATION

To overcome the problem of microbial contamination and its associated problems, the following

measures should be put in place.

1. Strict hygiene practice should be upheld in the processing and making of meat pie

through a thorough inspection to ensure compliance of standard.

2. Good manufacturing practice (GMP) should be observed within environment or space

used by meat pie vendors.

3. Meat pie should not displayed under the sun as the heat results to condensation thereby

allowing moisture accumulation which serves as a source of water to the potential

microbes.

4. Regulatory body such as national agency for food, drugs administration and control

(NAFDAC) and their counterparts in the local government area (sanitary/health

inspectors) should monitor all food products to ensure compliance of standar

21
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