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ABSTRACT

The effect of seasons on aflatoxins loads of selected pig’s feedstuffs in the humid tropics was
investigated, the feedstuffs sampled were cassava peels, bambara nut waste, palm kernel cake and
brewer spent grains. The study lasted for thirty-two weeks. These feedstuffs were collected in two
piggery farms in each of the six LGAs that make up Nsukka zone of Enugu State, in both dry and
rainy seasons.The collected feedstuffsamples were analyzed in the laboratory for aflatoxins
concentration levels using AOAC method of thin-layer chromatography (TLC)with some
modifications. Data were collected from the aflatoxins laboratory analysis of these feedstuffs and
with a structured questionnaire.The results of this study indicated the presence of aflatoxins in all the
sampled feedstuffs, however the lowest concentration of aflatoxins which is 0.011ppb is obtained in
cassava peels and bambara nut wastes while the highest concentration of aflatoxins obtained is
0.055ppb in brewer spent grains,these values are within the tolerance limit in Nigeria for total
aflatoxins (B1+B2+G1+G2),which is 20ppb.The obtained aflatoxins levels are within range and poses
less risk to pigs and pork consumers.The results also shows that there were no significant difference
(P > 0.05) on the aflatoxins concentration levels of the feedstuffs at their sources, across the piggery
farm locations, in the season and source of feeds interactions, season and the piggery farm locations
interactions, as well as season and feedstuffs interactions. However the main effect of seasonon
aflatoxins level which is 0.022ppb and 0.044ppb in the dry and rainy seasonrespectively were highly
significant (P < 0.01).Also the main effect of aflatoxins of the feedstuffs were highly significant (P <
0.01) except in brewer spent grains were it is significant (P < 0.05). It follows that the aflatoxins
levels according to season in the comparison between the source of feedstuffs and the piggery farm
locations for the sampled feedstuffs were highly significant (P< 0.01) especially in the rainy season,
except bambara nut wastewhich is highly significant (P< 0.01) in the dry season. There were higher
values in the aflatoxins levels in all the feedstuffs sampled in the rainy season than in the dry
season.It was therefore concluded that season has an effects on the aflatoxins level of pig’s feedstuffs
with rainy season favouring toxigenic mould metabolism and growth in the feedstuffs which
produces aflatoxins as their metabolites than in the dry season in this study, hence adequate
preservative measures, proper feed handling and storage should be adopted. It is therefore
recommended that piggery farmers should adopt adequate preventive measures and also apply the
various preservative storage methods in order to reduce the aflatoxins levels infeedstuffs especially in
the rainy season for better performance of the pigs and safety of pork consumers.

 

 

TABLE OF CONTENTS

itle Page … … … … … … … … … …i
Certification … … … … … … … … … …ii
Dedication … … … … … … … …. … …iii
Acknowledgement … … … … … … …. … …iv
Abstract … … … … … … … … … …v
Table of Contentss … … … … … … … … …vi
List of Tables … … … … … … … … …xi
List of Figures … … … … … … … … … ….xii
Chapter One
1.0 Introduction
1.1 Background of the study … … … … … … …….1
1.2 Statement of Problem … … … … … … …….4
1.3 Objectives of the study … … … … … … …. ……5
1.4 Justification of the study … … … … … … ……6
Chapter Two
2.0 Literature Review
2.1 Brief History of Aflatoxins … … … … … … … ……7
2.2 Natural Occurrence of Aflatoxins … … … … … ……8
2.3 Mycotoxins and Its Effects…. … … … … … …..10
2.4 Chemical Reactions of Aflatoxins … … … … … ….12
2.4.1. Heat …. … … … … … … … … ….12
2.4.2 Alkalis … … … … … … … … ….12
2.4.3 Acids … … … … … … … … … ….14
2.4.4 Oxidizing Agents … …. ….. … … … … ….14
2.4.5 Reduction … ….. …… … … … … …14
2.5 Chemical Structure of the Different Types of Aflatoxins … …15
2.6 Aflatoxin B1 Pathways … … … … … … …16
2.7 Metabolism of Aflatoxins … … … … … …16
2.7.1 Bio-activation … … … … … … … …16
2.7.2 Conjugation… … … … … … … … …16
2.7.3 Deconjugation … … … … … … … …17
2.7.4 Aflatoxins Units and its Conversion … …. … … …17
2.8 Factors that favours Mould Growth and Development … …18
2.9 Clinical Signs/Symptoms of Aflatoxicosis in Pigs… … … …18
2.9.1 Acute Aflatoxicosis… … … … … … …19
2.9.2 Chronic Aflatoxicosis … … … … … … …19
2.10 The Effects of Aflatoxins in Pigs … … … … …20
2.11 Aflatoxins Tolerance Levels and Action Limits in Animal Feeds …23
2.12 Aflatoxins and Climate Change… … … … … …27
2.13 Aflatoxins and its Economic Importance … … … …28
2.14 Aflatoxins Metabolism in Animals …. … … … …29
2.15 Methods of Aflatoxins Determination and Quantificationin Samples…. 30
2.16 AflatoxinsPrevention and Control Strategies…… … … ….32
2.17. Alternative Feedstuffs and their Uses in Pig Feeding … … …41
2.18 Cassava Peels Meal (CPM)…. …. … … … … …42
2.19.1 Nutritive Value of Cassava Roots/Peels … … … …44
2.19.2 Utilization ofCassava Peels as feedstuffs … … … ..… …45
2.19.3 Constraints to the Use of Cassava Meal/Peels as a feedstuff… … …47
2.20 Palm Kernel Cake (PKC)… … … … … … …48
2.20.1 Nutritive Value of Palm Kernel Cake as feedstuffs … … …49
2.20.2 Utilization of Palm Kernel Cake … … … … … …51
2.20.3 Constraints to the Use of Palm Kernel Cake … … … …52
2.21 Bambara Nut Waste (BNW)… … … … … … …53
2.21.1 Nutritive Value of Bambara Nut/Waste as feedstuff … … …54
2.21.2 Utilization of Bambara Nut /Waste .. … … … …56
2.22 Brewer’s Spent Grain (BSG)… … … … … …57
2.22.1 Nutritive Value of Brewer’s Spent Grains… … … … …58
2.22.2 Utilization of Brewer Spent Grains as feedstuff…… … … …61
2.22.3 Constraint to the Use of Brewer Spent Grains … … … …62
2.23 Brewer Yeast … … …. … … … … …63
2.23.1 Nutritive Value and utilization of Brewer Yeast in Livestock feeds… …63
2.24 Effect of High Content of Fibre in Pigs Diets … … … …65
2.25 Anti-nutritional factors in Non-conventional Feedstuffs …… …66
2.25.1 Classification of Anti-nutritional Factors (ANFs) … … …67
2.25.2 Factors Interfering with the Digestion and Utilization of Dietary
Proteins and Carbohydrates … … … … … …68
2.25.2.1 Tannins … … … … … … … …68
2.25.2.2 Saponins … … … … … … … …69
2.25.2.3 Trypsin or Protease Inhibitors…. … … … … …69
2.25.2.4 Haemagglutinins… … … … …. … … …70
2.6 Factors that Interfers with availability of Minerals … … …71
2.26.1 Phytates … … … … … … … …71
2.26.2 Oxalic Acid … … … … … … … …71
2.26.3 Glucosinolates… … … … … … … …71
2.26.4 Gossypol … … … … … … … …72
2.26.5 Phytoestrogens … … … … … … …72
2.26.6 Anti Vitamins … … … … … … … …72
2.26.7 Cynogens … … … … … … … …72
2.26.8 Lathyrogens … … … … … … … …73
2.26.9 Nitrates and Nitrite … … … … … … …73
2.26.10 Flatulence Factors … … … … … … …73
2.27 Pigs and its Characteristics … … … … … …73
2.28 Humid Tropics and its Characteristics … … … …74
Chapter Three
3.0 Materials and Methods
3.1 Location of the Study… … … … … … … … …76
3.2 Experimental Materials… … … … … … … … …76
3.3 Experimental Procedure… … … … … … … … …77
3.4 Sample Preparation … … … … … … … … …77
3.5 Procedure for Aflatoxins Determination and Quantification by (TLC)… …. …78
3.5.1 Apparatus… … … … … … … … … …78
3.5.2 Reagents… … … …. …. … … … … … …78
3.5.3 Procedure of AOAC Aflatoxins Analysis using TLC … … … … ….79
3.5.4 Method of Determination of Aflatoxins Concentration in Feedstuffs Samples … …81
3.5.5 Safety Issues… …. …. … … … … … … …81
3.6 Experimental Design… …. … …. …. …. … … …82
3.7 Data Collection… … … … … … … … … …82
3.8 Statistical Analysis … … … … … … … … …83
Chapter Four
4.0 Results and Discussion
4.1 Results and Discussion of Aflatoxins Laboratory Analysis of Pigs feedstuffs … …84
4.1.1: The results of the effect of source of feedstuffs on the aflatoxins levels of pig feedstuffs 84
4.1.2: The results of the effect of farm location on the aflatoxin levels of pig feedstuffs … 85
4.1.3: The results of the effect of season on the aflatoxins levels of pig feedstuffs …. 86
4.1.4: The results of the effect of feedtype on aflatoxins levels of pig feedstuffs …. …. 86
4.1.5: The results of the effect of season on aflatoxins levels of pig feedstuffs …. …. 87
4.1.6: The results of the effect of source and season on aflatoxins levels of pig feedstuff … 88
4.1.7: The results of the effect of feedstuff and season on aflatoxins levels of pig feedstuffs 89
4.1.8: The results of the effect of season and farm location on aflatoxins levels of feedstuffs 90
4.1.9: The results of the effect of source and farm location on aflatoxins levels of feedstuffs 91
4.2 Pig Farmers Responses and Discussions … … … … … … 95
4.2.1 Marital status … …. …. …. … … … … …. … 98
4.2.2 Educational Qualifications … …. … … … … … … 98
4.2.3 Gender… … … … … … … … … … … 98
4.2.4 Number of pigs reared in the farm … … … …. …. … … 99
4.2.5 Frequency of sourcing pigs’ feedstuffs …. … …. …. … …. 99
4.2.6 Quantity of feedstuffs purchased in kg/tons …. … …. …. … …. 100
4.2.7 How these feedstuffs are stored in the piggery farms…. …. …. … …. 100
4.2.8 Susceptibility of pig breeds to mould contaminated feedstuffs …. … …. 100
4.2.9 Ingredients added to the feedstuffs to improve its nutrient availability … …. 101
4.2.10 Mould incidence/infestation of feedstuffs fed to pigs…. …. …. …. …. 101
4.2.11 Feedstuffs that develop moulds faster than others …. …. …. …. …. 101
4.2.12 How does these symptoms affect the pigs … …. …. …. …. …. 102
4.2.13 Feedstuffs used by pig farmers to feed pigs in their farms …. …. …. …. 103
4.2.14 Main source of feedstuffs from where the pig farmers procure feedstuffs …. …. 104
4.2.15 Signs and symptoms shown by pigs when fed mould contaminated feeds …. …. 105
4.2.16 Preventive measures adopted to reduce mould infestation of the feedstuffs …. …. 106
4.2.17 The season/ period of the year mould infestation of feedstuffs are prevalent …. 107
Chapter Five
5.0 Summary,Conclusion and Recommendation
5.1 Summary … … … … … … … … … …109
5.2 Conclusion… … … … … … … … … …110
5.3 Recommendations … …. … … … … … …110
References
Appendices

 

 

CHAPTER ONE

INTRODUCTION
1.1 Background of the Study
The consumption of animal protein is very essential for meeting the protein requirement of man for
normal growth and healthy development. The protein sources from animal origin are good sources of
lysine and sulphur amino acids, which are lower in protein sources from plant origin (Omole, 1991).
The shortage of protein, particularly animal protein is prevalent in most parts of Africa where it is
estimated that on the average 10 grams of animal protein is consumed per day compared to the
recommended daily intake of 35 grams (ILCA,1980; FAO, 1986). The level of animal protein
consumption was estimated at 8 grams per caput per day, this is about 20 grams less than the
National Research Council of United States of America’s recommendation (Obioha, 1992; FAO,
1997). Ogini (2001) stated that Nigeria, despite its vast land area of 9,237,680km2 , the greater
percentage of her citizens of over 120 million are still battling with abject poverty and hunger. Atsu
(2002) reported that there is inadequate intake of animal protein in Nigeria.
The high cost of feeds and feed ingredients, especially the conventional energy and protein feed
sources such as maize, wheat, sorghum, soybean cake, groundnut cake and fish meal were as a result
of the stiff competition between human consumers for food, for livestock feeds/feeding and industrial
raw materials e.g. cereal grains for baby food formulas and biofuel production, especially maize
(Agbede et al., 2002; Adegbola and Asaolu, 2008). Maize is a major source of energy in monogastric
feeds, especially pigs, poultry and rabbits. In Nigeria, cereal grains are in high demand and its
production has never been adequate to meet the needs of the increasing population; consequently
there is little or no excess grains for livestock feeding. When available, it is always very expensive
(Rhule,1999). The cost of conventional feedstuffs which are major sources of energy and protein in
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monogastrics diets has continued to increase (Onu and Madubuike, 2006; Defang et al., 2008) due to
their short supply. In 2005 the price of maize rose from about N30 to N70 per kilogram
(Rhule,1999). As the competition between man, farm animals and industrial uses for grains continues
to increase, substitutes for grains are being increasingly used in poultry and livestock feeding to
reduce the high cost of feed (Babatunde and Hamzat, 2005). This continuous increase in the cost of
conventional feed ingredients used in formulating poultry and livestock feeds has necessitated an
intensive search for cheap alternative feed ingredients that will be able to replace a given portion, if
not all of the conventional feed ingredients (Babatunde, 1985; Onyimonyi, 2002; Adesehinwa et al.,
2011).
According to Ugwu et al. (2008), the main factors militating against the rapid expansion of animal
production industry in Nigeria is the problem of inadequate supplies of feedstuffs at economic prices.
The scarcity and high cost of conventional feedstuffs is largely responsible for the present high price
of finished animal products such as eggs, meat and milk (Rhule,1999; Adesehinwa et al., 2011).
Ijaiya et al. (2004) pointed out that feed cost is perhaps the most expensive input in intensively reared
stocks and constitutes about 70 – 80% of the real cost of animal production. Onyimonyi and
Onukwufor (2003) had earlier reported that a major solution to the problem of rising costs, scarcity
of energy and protein sources for monogastrics is to seek for new and non-conventional feed
resources which are able to replace a certain percentage proportion of the known conventional
feedstuffs without any deleterious effect on the animal performance. Esonu et al. (2001) indicated
that such substitute which have been successfully used to replace the conventional feedstuffs are
brewer spent grains, stem/tuber residues and cereal by-products such as wheat offal, rice bran, maize
bran as well as bambara nut waste and cassava or yam peels.
Fetuga et al. (1977), Iyayi et al. (2005) and Onyimonyi and Ugwu (2007) had studied the feeding
value of such non-conventional feed resources as palm kernel cake, bambara nut offal, rice husk and
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cassava peels, and that they can be utilized in supplementing a given portion of the conventional
feedstuffs. Onyimonyi and Ugwu (2007) further noted that these non-conventional feeding stuffs are
commonly found in the South Eastern Nigeria. According to Babatunde and Hamzat (2005), these
alternative feedstuffs have proved valuable in supporting the performance of livestocks and poultry
and that by utilizing them in ration formulation, it will go a long way in reducing the demand and
cost of conventional feedstuffs, which greatly increases the total cost of pork production. Adegbola
and Oduozo (1992) reported that some of the limiting factors associated with agro-industrial byproducts
and crop residues utilization as animal feeds include: procurement, storage, poor feed
intake, high fibre content, toxic and anti-nutrients substances as well as being low in their
digestibility which subsequently lowers the animal performance.
According to Serres (1992) pigs are known to be highly prolific and very efficient in converting feed
nutrients into high quality animal protein. Adesehinwa (2008) stated that pigs are used to convert a
variety of feeds into meat for human consumption. The supply of animal protein by pigs is limited by
many factors among which are inadequate nutrition and feeding; poor breeding stocks, poor
management practices and housing; disease prevention and control in pig production (Holness,1999).
Pitt and Miscamble (1995) defined mycotoxins as toxic secondary metabolites produced by microorganisms
of the fungus kingdom commonly known as moulds. Awan (2001) reported the ability of
moulds to produce toxins potentially dangerous to man and animals are common and widespread.
Odoemela and Osu, (2008) reported that toxigenic moulds have been found during growth, harvest
and storage of diferent foods and feeds and other agricultural produce. Abarca et al.(2001) stated that
there are five major agriculturally important fungal toxins of economic importance namely;
aflatoxins, deoxynivalenol, ochratoxin, zearalenone and fumunism, which are produced by fungi
invasion of agricultural produces and feed ingredients under favourable conditions of temperature
(25 – 30oc) and relative humidity of above 80% (Gilbert and Vargas, 2003). According to Farombi
4
(2006), these moulds contaminate a large number of dietary staples and agricultural produce such as
rice, corn, cassava, peanuts and spices, while noting that humans and animals are exposed to
aflatoxins by consuming contaminated foods and feeds. These fungal metabolites which when
ingested, inhaled or absorbed through the skin causes lowered performance, sickness or eventually
death in both animals and humans (Kuilman et al.,1998; CAST, 2003).
Atanda et al.(2008) stated that several moulds and yeast species were isolated from grains with
Fusarium verticilloides and Aspergillus flavus having the highest frequency of occurrence of 39.1%
and 22.3%, respectively. Moulds have the potentials of forming aflatoxins and trichothecenes in
grains, peanuts, cottonseed and other agricultural products (Atanda et al., 2008).
Adam (2000) reported that Aspergillus spp produces aflatoxins. According to Sashidhare et
al.(1992), the high incidence of mycotoxins contamination could be due to physical status of grains,
the moisture content, temperature, oxygen and the amount of carbon (iv) oxide in the atmosphere.
These factors are reported to influence the rate of infestation and proliferation of moulds in
agricultural produce and grains, especially under storage condition.
1.2 Statement of the Problem
Adesehinwa et al. (2008) stated that the use of alternative feedstuffs such as cassava peels, brewer
spent grains, palm kernel cake and bambara nut waste in feeding pigs is a way of reducing the cost of
pork production. These feedstuffs are procured and stored for a given period of time with different
handling and storage techniques before feeding them to the pigs. Within this period of storage and
handling, the feedstuffs may be exposed to mould infestation and the resultant aflatoxins
contamination (Osweiler, 2006). Henry (2009) stated that poor handling and storage of feedstuffs
adversely affect the quality and the nutrients supplied by the feedstuffs to the animals (Akbar and
5
Majid, 2010). Pig farmers may be feeding their pigs with mould contaminated feedstuffs without
knowing that such feedstuffs are contaminated
Aflatoxicosis in pigs reduces feed intake, reduce weight gain, reduce growth rate,
immunosuppressive including antibody and interleukin production, causes liver damage and
hemorrhage (Creepy, 2002; Carvajah et al., 2003; Sun and Chen, 2003; Lu, 2003). Pitt and
Miscamble (1995) indicated that the accumulation of aflatoxins, which is a major mycotoxins in the
feedstuffs used in feeding animals poses a serious threat to animal and human health, pointing out
that they are responsible for many different toxicities and diseases such as cancer, vascular, kidney,
nervous disorders and eventually death of the animals or persons consuming such animal produce
(WHO 1979; Kuilman et al., 1998; CAST, 2003). Hussein and Brasel (2001) and Akbar and Majid
(2010) stated that aflatoxins B1, B2, G1 and G2 are toxic substances, although aflatoxin B1 is the most
frequent and toxic of the aflatoxins group (WHO, 1979; Pier, 1992). These toxins are wildly
implicated in causing health risks to human due to its carcinogenic, mutagenic and teratogenic
properties in persons consuming pork products that are high in aflatoxins concentration over time
(WHO, 1979; Kuilman et al., 1998; Sun and Chen, 2003).
1.3 Objectives of the Study
The broad objective of the study was to determine the effect of season on aflatoxin load of selected
feedstuffs for pigs in the humid tropics. The specific objectives were
i. To determine the level of aflatoxins in pig feeding stuffs
ii. To determine the level of aflatoxins of stored feedstuffs at the piggery farms
iii. To determine the effect of seasons on the aflatoxins levels in the sampled feedstuffs
iv. To compare the levels of aflatoxins in feedstuffs at source and stored in the piggery farm
v. To determine pig farmers perception of aflatoxins and their implications
6
1.4 Justification of the Study
This study is necessary in order to identify the level of aflatoxins in the various feedstuffs used in
feeding pigs in the humid tropics. Tewe and Adesehinwa (1995) and Ijaiya et al. (2004) stated that
feeds constitute over 70% of the cost of pig production. These feeds are rendered poisonous with
deleterious effects by aflatoxins contamination as a result of mould infestation when fed to the pigs.
The deleterious nature of contaminated feeds/feedstuffs (Pier, 1992; Odoemelam and Osu, 2008) on
animal health, predisposes the pigs to diseases by reducing feed intake, lowering their immune
system, resulting in low performance and death, especially in piglets (Sun and Chen, 2003; Lu,
2003). It also increases the total cost of pig production in terms of slow maturity/growth rate, high
cost of medication/treatment of affected pigs and death (CAST, 2003). Pork consumers are also at
health risks with the exposure and accumulation of aflatoxins in pork products of pigs consuming
such feeds (Hussein and Brasel, 2001; Ozsoy et al., 2005; Akbar and Majid, 2010). This could cause
cancer and other systemic diseases in human (WHO 1979; CAST 2003). This study is necessary to
assess the aflatoxins levels of pig’s feedstuffs and pig farmers awareness of its implications in pig
production. Aflatoxicosis outbreaks have been reported in Kenya, Uganda, China and India in recent
times (Lanyasunya et al., 2005). According to Pestka et al. (1999), stated that mycotoxin especially
aflatoxins contamination of diets can result in poor production and financial losses. Hence the
findings from this study will give an appraisal of the level of exposure of pigs and pork consumers
to aflatoxins. The results will also help pig farmers to adopt measures to minimize the level of
aflatoxins in pig’s feedstuffs and its implications. It will also increase pig farmers income, by
reducing the cost of pig production in the sampled areas.

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