Studies on combined effect of Aeromonas hydrophila and cadmium on lipid per...
EVALUATION OF THE EFFECT OF Moringa oleifera MARINADE ON CHORKOR SMOKED FISH
1. 1
EVALUATION OF THE EFFECT OF Moringa oleifera
MARINADE ON CHORKOR SMOKED FISH
(Clarias gariepinus) IN CROSS RIVER STATE
BY
---------------------------
SUBMITTED TO
DEPARTMENT OF ZOOLOGY AND
ENVIRONMENTAL
BIOLOGY (MARINE BIOLOGY UNIT)
UNIVERSITY OF CALABAR, CALABAR
2. 2
IN PARTAL FULFILLMENT OF THE COURSE
REQUIREMENT FOR THE AWARD OF BACHELOR
OF SCIENCE DEGREE IN MARINE BIOLOGY (B.SC
MARINE BIOLOGY)
MAY, 2015.
CERTIFICATION
I, ---------- with Registration Number -------- hereby certify
that this project entitled “Effect of Moringa oleifera marinade on
Chorkor Smoked fish (Clarias gariepinus) in Cross River State”
is original and written by me, it is a record of the research work
and has not been presented before on any previous publication.
9. 9
ABSTRACT
The study assessed the effect of Moringa oleifera marinade on
Chorkor Smoked fish (Clarias gariepinus). Thirty (30) African
catfish where shared into six (6) groups of 5 fishes each. Group
1 served as control, and group 2 was treated with 1% Moringa
oleifera Marinade, group 3 was treated with 2%Moringa oleifera
Marinade, group 4 was treated with 3% Moringa oleifera
Marinade, group 5 was treated with 5% Brine and group 6 was
treated with 0.2% Butylated hydroxyl anisole (BHA) (W/V)
solution. Thirty fishes of average weight of 230±8g were gutted
washed and randomly assigned to the treatment. Thereafter the
fishes were soaked in the treatment for 2 hours and later hot
smoked in chorkor for 12 hours. After smoking the fish were
stored in netted boxes and placed on shelves at room
temperature of (37±3
0
C) for 3weeks. Samples were taken every
three days for moisture content, colour, flavour and taste
analysis. The result showed that, 3% of Moringa oleifera
Marinade sample had the least moisture content, and the
10. 10
moisture reducing ability is dependent on the quantity of
Moringa oleifera Marinade. The taste, colour and flavour scores
showed no significant difference (P>0.01) among the treated
samples. The Moringa oleifera Marinade gives the treated
smoked fish an attractive colour and flavor. Therefore, Moringa
oleifera Marinade should be used at 3% to obtain the best
quality attributes and increase shelf-life of chorkor smoked
African catfish.
11. 11
CHAPTER ONE
1.0 INTRODUCTION
Fish received increased attention as a potential source of
animal protein and essential nutrients for human diet
(Ekpenyong and Ibok, 2012). Maintenance of high quality fish
therefore call for adequate, effective and affordable
preservative techniques to enhance preservation of this protein
resources. Post harvest losses of fish may reach 35%; in some
cases are nearly 25million tones of the world’s catch and in
some developing countries, post harvest losses of fish exceed
those of any other commodity, often surpassing 50% of landed
catch (FAO, 2000). An estimated 40% of total fish landing in
12. 12
Nigeria is lost as post harvest losses (Eyo, 2001). Due to the
susceptibility of fish to chemical, microbial and physical
deterioration, various preservation techniques are put in place
to check spoilage (Eyo, 1992).
Fish spoilage is a metabolic process that causes fish to
be undesirable for human consumption due to changes in
sensory and nutritional characteristics. Thus processing and
preservation of fish were of utmost importance to maintain
product quality, reduce wastage and prevent economic losses
(Olley et al., 2000). These include chilling, freezing, salting,
canning, drying and smoking (Kumolu et al., 2010). However,
13. 13
smoking is the most popular method of fish processing in
Nigeria (Bako, 2005).
Fish smoking is particularly relevant in the artisanal
fisheries sector in that it prolongs the shelf-life of the fish,
enhance flavor and increase utilization of the fish in addition to
reducing waste as well as increasing protein availability to
people (Jallow, 1995). However, smoke-dried fish are liable to
microbial damage reduced shelf life. Microbial spoilage could
predispose consumers to health hazards resulting from food
poisoning (Gram et al., 2000). In addition, smoke deposited on
fish during smoking is composed of carboxyl and some
polynuclear aromatic hydrocarbons, which have been
14. 14
implicated in degenerative diseases such as cancer (Eyo,
2001). In order to curb fish spoilage, increase shelf life, add
value to products and avoid food poisoning due to chemical
anti-biotic insecticide and anitimicrobial of plant origin has been
the working force of the researchers for years, this gives credit
to Moringa oleifera due to it antibacterial, insecticidal and
microbial activity in accordance with Caceres et al., (1991), and
Daxenbichler et al., (2003). The objective of this study is to
determine the effect of Moringa oleifera on Chorkor smoked fish
(Clarias gariepinus) in Cross River State.
15. 15
1.1 Aims and Objectives
This study is aim at “Effect of Moringa oleifera marinade
on Chorkor Smoked fish (Clarias gariepinus).”
Particularly, the studies aimed to;
a. Assess the effect of Moringa oleifera marinade on shelf-
life of Chorkor Smoked fish (Clarias gariepinus)
b. Compare the efficacy of the marinade to other
conventional treatment on the improvement of shelf-life of
chorkor smoke fish
16. 16
c. And to assess the marinade effect on the organoleptic
properties of the fish.
1.2 Justification
Synthetic Anisole and other dehydrating substances are
expensive and unaffordable by the common farmers in sub-
saharan Africa and in countries with transitional economies.
The side effects of these chemical have not been fully
understood. Therefore, complimentary alternative bio-
dehydrating practice has shown over the years that plant
contain potent bioactive dehydrating substances that safely
reduces moisture content and also improve proximate
17. 17
composition of the fish. This was justifiable for Moringa oleifera
by the natives as dehydrating substance to be scientifically
evaluated.
1.3 Scope of Study
This study was limited to a 7 week evaluation of the effect
of Moringa oleifera on marinade Chorkor Smoked fish (Clarias
gariepinus).
1.4 Research Hypothesis
Null Hypothesis
The administration of the Moringa oleifera marinade have
effect on the chorkor smoked fish (Clarias gariepinus).
19. 19
CHAPTER TWO
LITERATURE REVIEW
2.0 PLANT OF STUDY (Moringa oleifera)
2.1. Taxonomic Classification
Kingdom - Plantae
Sub-kingdom - Tracheobinta
Super division - Spermatophyta
Division - Magnoliophyta
Class - Magnoliopsida
Sub-class - Dilleniidae
Order - Capparales
Family - Moringaceae
Genus - Moringa
Species - Oleifera
20. 20
(Source; Garima et al., 2011)
2.1.2 Geographical Distribution
The tree is wild in the sub-Himalayan tracts from
Chenab to Oudh. It grows at elevation from sea level to
1400m. It is commonly cultivated near houses in
Assam, Bengal and Peninsular India. It is also a prolific
copier (Gupta, 2010). It is also cultivated in north–
eastern Pakistan, north-eastern Bangladesh, Sri Lanka,
West India and Southern Florida in central and South
America from Mexico to Peru as well as in Brazil and
Paraguay (Roloff et al., 2009).
2.1.3 Plant Morphology
Moringa oleifera is a small, fast growing evergreen
or deciduous tree that usually grows as myth as 9m
with a soft and white and corky and gummy bark. Root
have taste of horse-radish leaves are longitudinally
21. 21
cracked leaves, 30-75cm long main axis and its branch
jointed, glandular at joints, leaflet are glabrous and
entire. The leaflets are finely hairy, green and almost
hairless beneath with red-tinged mid-veins, with entire
margins and are rounded or blunt-pointed at the apex
and short pointed at the base. The twigs are finely
hairy and green. Flowers are white, scented in large
auxiliary down particle pods are pendulous, ribbed,
seeds are 3-angled (Gupta, 2010; Roloff et al., 2009).
2.1.4 Nutritional Composition of Moringa oleifera
According to Fuglie (2005), Moringa oleifera plant
forms the basis for several nutritional programmes in
many poor countries by charitable organization given
that the leaves of Moringa oleifera tree are rich is
essential nutrients.
22. 22
The leaves of Moringa oleifera are considered to
give immense possibilities for those who are
nutritionally challenged and may be regarded as a
protein or calcium supplement (Rajangan et al., 2001).
Bamishanye et al., (2011) reported Moringa oleifera
leaves of all stages having varying percentages of
nutritional composition. Fruglie (1999) was quick to say
the “so good to be called syndrome was the biggest
challenge for moringa”. Bureau of plant industry
reported Moringa oleifera as an outstanding source of
nutritional components. Its leaves (weight per weight)
have the calcium equivalent of four times that of milk,
the vitamin C content is seven times that of oranges,
while its potassium is three times that of bananas,
three times the iron of spinach, four times the amount
of vitamins A in carrots, and two times the protein in
milk (Kamal, 2008). In addition, the leaves can serve
23. 23
as a rich source of beta-carotene Nambiar and
Seshadri, 2001), Vitamin C and E and polyphenolics
(Ross, 1999). Also moringa is suggested as a viable
supplement of dietary minerals. The pods and leaves of
moringa contains high amount of Ca, Mg, K, Mn, P, Zn,
Na, Cu and Fe (Aslam et al., 2005). In addition, six
tablespoon full of Moringa oleifera leaf powder will
provide nearly the woman`s daily iron and calcium
during pregnancy and breast feeding hence Moringa
oleifera has been used to combat malnutrition among
infants and nursing mothers (TFL, Zoll) and the present
of it high protein and carbohydrate content makes it
suitable for treatment of protein energy malnutrition
problem (Williams et al., 2013).
2.1.5 Phytochemical Composition of Moringa
oleifera
24. 24
Moringa and lemmen (2007) reported the isolation
of five flauonol phycosides characterized as
kaempferide3-0-(2’,3’–diacetylglucoside), kaempferide-
3-0-(2”-0-galloylrhamnoside), kaempferide -3-0-(2”-0-
galloylrutriside) 1-7-0-alpha-rhamnoside, kaempefrol-
3-0-(beta-glucosyl-(1 - 2). [alpha-rhamnosyl-(1 - 4).
Betaglucoside-7-0-alpha-rhamnoside together with
benzoic acid 4-0-beta-glucoside, benzoic acid 4-0-
alpha-rhamnosyl-(1 - 4). Betaglucoside-7-0-
alpharhamnoside together with bezoic acid 4-0-
betaglucoside, benzoic acid 4-0- alpharhamnosyl- (1 -
2) betaglucoside and benzaldehyde 4-0-betagluside
have been isolated from methanolic extract of MO
leaves. Other reports include the presence of niazirin,
niazirin, three oil mustard oil glycoside; 4—[(4-0-
acetyl-alpha-l-rhamnosyloxy) benzyl] isothiocyanate,
niaziminin A and niaziminin B (Faizi et al., 1994).
25. 25
Phytochemical studies on Moringa oleifera by M.
Ndong et al., (2007) revealed major polyphenols such
as quercetin glucosides, rutin, kaempterol ghycosides
and chlorogenic acid in Moringa oleifera powder.
Manguro and lemmen (2007) reported the
isolation of flauonol phycosides characterized as
syringic acid, gallic acid, rutin and generation. It leaves
also contain same amino acid such as aspartic acid,
glutamic acid, glycine, threomine, alanine, value,
leucine, isolencine, hostidine, lysine, phemlalamine
tryptophan, cysteine and methionine (Ram and
Mehrotra, 2006).
2.1.6 Pharmacological Actions of Moringa oleifera
2.1.6.1 Antimicrobial Activity
Caceres et al., (1991) reported antimicrobial
activities of Moringa oleifera leaves, roots, bark and
26. 26
seeds in vitro against bacteria, yeast, dermatophytes
and helminthes by a disk-diffusion method. The juice
and aqueous extracts from the seeds inhibited the
growth of pseudomonas aeruginsa and staphylococcus
aureus. Also Hueih-min chen et al.,(2007) reported
invite antifungal activity from the ethanolic extract of
the leaves of Moringa oleifera against dermatophytes,
Epidermophyton xoccosum and microsyonum canis.
Amer Jamil et al., (2008) evaluated antimicrobial
activity from the seeds of Moringa oleifera. The seed
extracts shows antimicrobial activity against bacterial
(pasturella multocide, Escherichia coli, Baccillius
substile and Staphylococcus aureus) and fungal
(Fusarium solani and Rhizopus solani) strains.
In addition, Nickon et al.,(2003) reported
antimicrobial activity of aglycone of Deoxy-Niazimicine
27. 27
which is characterized as N-benyl, s-ethyl thioformate
from the chloroform extract of Moringa oleifera root
barks. The compound shows antibacterial and
antifungal activities against shizella boydi, and
staphylococcus aureus.
2.1.6.2 Anti-inflammatory Activity
Mahajan et al.,(2007) report anti-inflammatory
activity of ethanolic extract of seeds of Moringa oleifera
against immune mediated inflammatory responses in
toluene disocyanate- induced asthma in wistar rats.
Aurantiamide acetate 4 and 1,3 dibenyl urea 5 isolated from the
roots of MO was reported to inhibit the production of TNF-alpha
and IL-z (Sashidhara et al, 2009).
28. 28
Mahajan et al. (2007) reported anti-arthritic activity of
ethanolic extract of seeds of Moringa oleifera. In adjuvant-
induced arthritis in adult female wistar rats. In addition he also
reported the anti-inflammatory activity from the a-butanol
extract of seeds of Moringa oleifera against ovalbumin-
induced airway inflammation in guinea pigs (Mahajin, et al.,
2009).
2.1.6.3 Antioxidant Activity of Moringa oleifera
Epidemiological studies have shown that goods rich in
vitamins provide protection against degenerative diseases
including cancer, cornay heart disease and even Alzheimersis
29. 29
disease (Ames et al, 1993). Plant containing antioxidants like
vitamin C, vitamin E, carotenes, pohyphwnola and many other
compounds reduce these disease risks. Most of the antioxidant
compounds in a typical balanced diet are derived from plant
sources with a wide variety of Biological and chemical
properties ( Scalbert et al, 2005).
Siddhuraju and Becker (2003) reported antioxidants and
radical scavenging property from water, aqueous methods and
aqueous ethanol extracts of freeze dried leaves of Moringa
oleifera. And th leaf extracts were capable of scavenging
peroxyl and superomyl radicals. Sreelatha and Padma (2009).
Evaluated antioxidant effects of Moringa oleifera laef extracts,
30. 30
which were tested in two stages of maturity using standard in
vitro models. The successive aqueous extract of Moringa
oleifera. Exhibited strong scavenging effect on 2,2-diphenyl-2-
picryl hydrazyl (DPPA) free radical, superomide, nitric oxide
radical and inhibition of lipid peroxidation.
Also Verma et al, (2009) reported invitro and in vivo
antioxidant properties of different fraction of leaves of MO using
different in vitro sperms, such as bet-carothene bleaching,
reducing power, DPPH/Superoxide/hydroxyl radical
scavenging, ferrous ion chelation and lipid peroxidation. Leaves
rich in biologically active carotenoids, tocopherols and vitamin
31. 31
C have health promoting potentials and prevent free redical
damage that can initiate many illness ( Smolin et al., 2007).
2.1.6.4 Anti-cancer Activity
Murakami et al., (1998) reported antitumor activity from the
leaves of Moringa oleifera. The antitumor potential was due
to the presence of three known thiocarbonate (TC) and
isothiocyenate (ITC) – related compounds which are acting as
inhibitor of tumor promoter teleocidin B-4- induced Epstein –
Bar Virus (EBV) activation in Ragi cells.
32. 32
Guevara et al., (1999) also reported anticancer activity
from the ethanolic extract of seeds of Mo. The seed extract was
found to contain various antitumor compounds like O-ethyl-4-
( alpha-L- rhamnosyloxy) benzyl carbonate which was a new
compound, together with seven known compounds, 4( alpha-L-
rhamnosyloxy) – benzyl isothiocyanate, niazimicin, niazirin,
beta-sitosterol, glycerol –L- (9- octadecanoate), 3-0- (6-0-
oleoyl-beta-D- glucopyranosyl) –beta- sitosterol and beta-
sitosterol -3-0- beta D- glucopyranoside. Their potential
antitumor promoting activity was performed using an in vitro
assay which tested their inhibitory effects on Epstein- Bar Virus-
early antigen (EBV-EA) activation in Raji cells induced by by
33. 33
the tumor promoter, 12-0- tetradecanoyl- phorbol-13- acetate
(TPA).
2.1.6.5 Cardiovascular Activity
Faizi et al. (1994) reported the isolation of two nitrile
glycosides from the ethanolic extracts of Moringa oleifera
leaves, niazirin and niazirinin and three mustard oil glycosides.
Compounds such as 4-[(4-0- acetyl- alpha-L- rhamnosyloxy)
benzyl] isothiocyanate, niaziminin A and B showed hypotensive
activity. He also reported six new synthetically known
glycosides from the ethanolic extract of the leaves of Moringa
oleifera. Most of these compounds, bearing thiocarbamate,
34. 34
carbamate or nitrile groups, are fully acetylated glycosides,
thiocarbamates showed hypotensive activity (Faizi et al., 1995).
Ghasi et al.(2000) investigated hypocholester olemic effect
of crude leaf extract of Moringa oleifera. The effect on the
serum cholesterol was statistically significant. And Ndong et al.
(2007) tested preventive effects of Moringa oleifera on
hyperlipidemia caused by iron deficiency in male wistar rates.
Nandave et al. (2009) investigated cardioprotective effect
of lyophilize hydralcoholic extract of Moringa oleifera in the
isoproterenol (isp) – induced model of myocardial infaction in
male wister albino rats. Chronic treatment with Moringa
35. 35
oleifera resulted in significant favorable modulation of the
biochemical enzymes (superoxide dismutase, catalase,
glutathione peroxidase, lactate delydrogenase, and creatine
kinase- MB). The leave extract also showed antiatherosclerotic
and hypolipidaemic activity (Chumark et al., 2008 ).
2.1.7 Traditional Uses of Moringa oleifera
The leaves, seeds, flowers, pods(fruit), and roots are
seen as a vegetable and each part is uniquely harvested and
utilized.
36. 36
a. Fresh leaves are picked, shade dried, ground to a
powder, and then stored for later as a food flavoring or
additives. Dried or fresh leaves are also used in food
such as soups and porridges (Lockett et al., 2000), curry
gravy and in noodles, rice or wheat (Albilgos et al., 1999).
b. Farmers have added the leaves to animal feed to
maintain a healthy livestock (Sarwatt et al., 2002; Fahey,
2005’ Sancheza et al., 2006) while utilizing the manure
and vegetables compost for crop growth (Fahey, 2005;
SaveGaia international Foundation, 2005).
37. 37
c. Newer applications include the use of Moringa oleifera
powder as a fish food in aqua cultural system (Dongmeza
et al., 2006) and the Moringa oleifera leaves as a
protein supplement for animals such as cows. The
feeding value of Moringa oleifera has been reported to
be similar to that of soyabeans and repeseed meal
(Soliva et al., 2005).
d. With the leaves being rich in nutrient, pregnant women
and lactating mothers use the powdered leaves to
enhance their child’s or children’s nourishment, especially
in under developed countries suffering from malnutrition
38. 38
(MC Burney et al., 2004; Lockett et al., 2000; WHO
Readers forum, 1999).
e. The whole seeds can also be eaten green, roasted or
powdered, and steamed in tea and curries (Fahey, 2005).
The pods and seeds, often referred to as Moringa
oleifera kernels, have a taste that ranges from sweet to
bitter and are most popularly consumed after frying to get
a peanut-like taste (Makkar et al.,1996).
TABLE 2.1.7. The Traditional Uses and Medical
Applications of Moringa oleifera
39. 39
PARTS
OF THE
TREE
USES TRADITIONAL
METHODS OF
PREPARATION
MEDICAL PURPOSES
Leaves Salads,
vegetables,
curries, powder
for scrubbing
utensils
Fresh or dried
leaves are grouped
to a powder and
use to prepare
salves
Treating tumors; as
poultice for spores,
reduce glandular swelling
and headaches, to purge
or a body cleanser, to
promote digestion;
traditional medicine as a
hypocholesterolemic
agent in obese
individuals.
Seeds Eaten as a
snack , oil for
salad, cooking,
cosmetics,
lubricant; water
purifying
Seeds are prepared
green ,roasted or
powdered, steamed
and extracted as an
oil.
Treats abdominal tumors ;
removes harmful
bacterials
Flowers For honey Flower extract are
used for
preparations
Folk remedies for tumors
Bark For tanning Decoctions for
creams or
emollients
Promote digestion
40. 40
The fish from within fish tissues and evaporates. The air
surrounding the fish the experiences a drop in temperature.
This is accompanied by cooling of the surface of the fish. The
energy required to drive the moisture from the surface of the
fish can be obtained from a variety of sources including wood
smoke, sun drying, solar drier electricity and mechanical driers
(Davis et al, 2008).
2.2 Animal of the study (Claria gariepinus)
2.2.1 Scientific of Claria gariepinus
Kingdom - Animalia
Phylum - Actinopterygil
Order - Siluriformes
41. 41
Family - Clariidae
Genus - Clarias
Species - C. gariepinus
2.2.2 Description of Claria gariepinus
The African sharp tooth cat fish is a large, eel-like
fish, usually of dark gray or black coloration on the
back, finding to a white belly. In Africa, this catfish has
been reported as being second in size only to the
Vundu of the Zamdesian waters, although fish base
suggests the African sharp tooth catfish surpasses that
species in both maximum length and weight (Forese et
al, 2014).
Claria gariepinus has an average adult length of 1-
1.5m (3ft 3in – 4ft 11in). it reaches a maximum length
of 1.7m (5ft 7in) TL and can weigh up to 60kg (130k)
42. 42
(Foreses et al, 2014). The fish have slender bodies, flat
bony heads notably flatter than in the genius silurns,
and broad terminal mouths with four Paris of barbels.
They also have large accessory breathing organs
composed of modified gill arches. Also, only the
pictorial fins have spines.
2.2.3 Fish Spoilage
Fish is an extremely perishable food item (Agbon
et al., 2002). So after death, fish begins to spoil. In the
healthy live fish, all of the complex biochemical
reactions are balanced and the fish flesh is sterile. After
death however, irreversible change that results in fish
spoilage begins to occur. The resultant effect is the
decomposition of the fish (Akinola et al., 2006). Various
factors are responsible for fish spoilage. The quality of
capture is important at determining the rate of
43. 43
spoilage. Notably, are the fishes health status, the
presence of parasites, bruises had wounds on the skin
and the mode by which the fish was captured. The
caught fish quality depends on the handling and
preservation; the handling and preservation practice
after capture affects the degree of spoilage of the fish
(Akinneye et al., 2007).
Fish spoilage is brought about mainly by, the
enzymes present in the live fish. The enzymes begin to
break down fish tissues. Prior to death, the enzymes
were involved in the digestion of ingested food and all
enzymatic reaction are controlled. In the death fish, the
control system fails and the enzymes begins to act on
the alimentary system and fish flesh, thereby remitting
in soft destructive changes. This process is referred to
as autolytic spoilage (FAO, 1985).
44. 44
Bacteria are present in the gut, gills and skin
surfaces of live fish. The live fish defense mechanism is
able to combat the action of these bacteria. However,
some after death, this defense mechanism also fails.
Consequently, the bacteria invade the gut, gills and
skin and cause the decomposition from within and the
exposed surfaces of the fish.
Enzymes and bacteria spoilage of fish can be
reduced or temporarily halted by various techniques.
The tradition and popular methods employed include;
a. Temperature reduction by use of ice (Freezing or
ice blasting)
b. Drying to reduce or completely remove water
c. Salting to reduce water and stop enzymatic
decomposition.
45. 45
d. Application of heat example; canning and smoking
to destroy the enzymes and kill all bacteria.
2.2.4 Fish handling
Fresh fish after captive should be properly handled
if keeping quality and shelf life are to be improved
reasonably (Anthonio, 1970). One of such good
handling practices is to ensure that captured live fish
are not allowed to struggle and die of asphyxia or
oxygen starvation. Struggling after capture except in
the case of the catfishes, will hasten spoilage post-
mortem by accelerating, chemical reactions in the flesh
of the fish. This will reduce the period the fish will
remain in rigor or stiffened thereby, accelerate bacteria
attack and spoilage. Catfishes are known to remain
alive for along time after capture and should not be
stunned (Ayuba and Omeji, 2006). Other species
46. 46
should not be demobilized by piecing the brain with a
sharp object or by giving a blow to the head to ensure
instant death. Fresh fish deteriorates very rapidly. It is
necessary to ensure that fish and fish products get to
the consumer in acceptable quality, the initial handling
of freshly caught fish prior to processing must fulfill
certain conditions to maintain the acceptable quality
(Azeza, 1979). It is mandatory that they should not be
walked over, instead, they should be kept in boxes, or
baskets at chilled temperature shielded from direct
sunlight. Fish deteriorate very rapidly in the tropics
especially when they have been glutted or filleted due
to the high temperature and this increased exposed
surfaces. These should be kept chilled to minimize
microbial spoilage (Davies et al., 2008).
Filleting: The process of filleting involves laying the
fish on one side and cutting from behind the base of
47. 47
the pectoral fin, surrounding the back of the head. The
cut portion is then extended towards the tail along the
backbone. The rib bones are freed from the flesh,
which is also carved of the skin muscle covering the
abdomen. The tail is then separated from the block of
flesh. The fish is the turned and the other side treated
in the same vein. This method if expertly handled
produces simple fillets (FAO/UN.1963)
Gutted Fish: is a fish from which the viscera have
been removed. This is achieved by cutting along the
vertical surface from the operation to the anus. The
intestines, viscera and gills are removed. The removal
of the lead is optional. The removal of the gut implies
that the store house of enzymes responsible for
autotypic spoilage has been removed. Cutting of fish is
a normal process. Large volume of water is required for
washing gutted fish. The water should be chlorinated
48. 48
and under slight pressure to blood reaching the bone
(FAO/UN, 1969).
2.2.5 Fish Processing/Preservation
Appropriate processing of fish enables maximal
use of raw material and production of value-added
products which is obviously the basis of processing
profitability. Freshwater fish processing, like the
processing of the other food raw materials should,
assure best possible market quality, provide a proper
form of semi-processed final product assured health
safety of products apply the most appropriate
processing method and reduce wastes to the barest
possible extent. Akinneye et al. (2007) and Davies
49. 49
(2005) reported that the development of appropriate
fishing machinery and techniques that employed
effective production, handling, harvesting, processing
and storage cannot be over-emphasized especially in
the age when agriculture development is fast gathering
momentum in Nigeria.
It is imperative to process and preserve some of
the fish caught in the period of abundance so as to
ensure an all year round supply. This will invariably
reduce post harvest losses, increase the shelf-life of
fish, and guarantee a sustainable supply of fish during
off season with concomitant increase in the profit of the
fisher folks. Akinola et al., (2006) reported different
types of preservation methods; drying, smoking,
freezing, chilling and brining. But the most prominent
fish preservation in Niger Delta is smoking drying. This
could be adduced to the fact that most of the fish
50. 50
communities have no access to electricity to freeze
their products.
Akinola et al., (2006) reported that despite the
rudimentary nature of process of traditional methods,
lack of control over the drying rate, sometimes results
to under-drying or over-drying, expose the fish to
unexpected winds, dust, dirt, insect infestation, and
contaminants such as flies. To reduce post harvest
losses and to improve the quality of fish and fishery
products, traditional processing technology must be
improved upon in Nigeria. This includes upgrading the
traditional fish processing technology and adoption of
solar dryers. Artificial dryers such as solar dryer, kiln,
oven and so on, have long been in existence, some of
them are powered electrically, by sun, gas or natural
fuel such as firewood, charcoal, wood and saw dust
(Bolaji, 2005; Olokor, 1997, Igbeka, 1986). Ayuba and
51. 51
Omeji (2006) reported that the insect infestation as the
cause of most prominent losses in quality and quantity
of stored, dried fish in Nigeria.
They are of two types:
a. Initially when the moisture content of the fish is
high, it provides suitable breeding ground for
several species of flies.
b. When the moisture content is low dermestes
beetles vanish the product causing severe loss.
2.2.6 Method of Fish Preservation
2.2.6.1 Chilling, supper chilling and freezing
Chilling: chilling may be defined as cooling of fish to
low temperature without necessarily hardening fish.
Chilling does not prevent spoilage. However, the colder
the fish the better and the lower are the incidences of
52. 52
microbial or enzymatic spoilage. Bacteria or enzyme
action are not completely stopped but they may be
temporarily halted by chilling. To chill fish, the fish has
to be surrounded by colder medium, which could be
solid such as ice or liquids such as refrigerated water
(Ita, 1973)
Super chilling: This is not a common method, super
chilling implies reducing the temperature of fish
uniformly below 00
c. At this temperature half the water
in the fish freezes, bacteria action is greatly reduced
and self-life is extended. Fish are initially chilled using
ice before storage in refrigerator holds at temperatures
below freezing ice. The temperature in the hold is
maintained by means of cold or circulating refrigerated
brine. This method is known to extend shelf life of fish
by up to 4days (Ita, 1972).
53. 53
Freezing: Freezing is distinct from chilling of fish.
Freezing can keep products in near perfect conditions
for very prolonged periods freezing is essential for
expo. A purpose freezing becomes extremely effective,
if it is combined with cold storage (Anthonio, 1970).
Fish that have to be preserved by freezing should
be cleaned and packed before rigor mortis sets in for
easy operation and maximum use of freezing space.
Fresh fish have a characteristic sweet flavor, which is
due in part to inosinic acid. The breakdown of inosinic
acid during autolytic spoilage resulting in the
production of hypoxanthine results in the loss of the
sweet flavor to bitter flavor. Sugar is produced by
enzymatic action, which in turn reacts with the amino
acids to produce the brownish or yellowish color found
in frozen fish (Azeza, 1976).
54. 54
2.2.6.2 Drying: Drying is defined as the removal of
water by evaporation, when applied to fish, drying is
the removal of water by any method as a means of fish
preservation to prolong the shelf-life. In areas where
sun drying is used traditionally, the effects of wind and
weather conditions are important. Basically, the drying
effect of the sun depends on the emission of heat from
the sun. This transferred to the fish and, it is
accompanied by heat transfer within the fish. During
drying, the fish shrinks and undergoes irreversible
changes. Water is removed from the surface in the
following sequence. Firstly, water on the surface of the
fish evaporates. Water migrates to the surface of the
fish from within fish tissues and evaporate. The air
surrounding the fish then experiences a drop in
temperature. This is accompanied by cooling of the
surface of the fish. the energy required to drive the
55. 55
moisture from the surface of the fish can be obtained
from a variety of sources including wood smoke, sun
drying, solar drier, electricity and mechanical driers
(Davies et al., 2008).
2.2.6.3 Smoking
Smoking is a popular traditional method of fish
preservation in most developing countries. Smoking combines
the effect of the destruction of bacteria by compounds in the
smoke, such as phenols and the cooking of the fish since, high
temperatures will be generated. Smoked fish products have
long shelf-life, which has been attributed to the drying and
cooking effects. When wood and sawdust are burnt, smoke is
produced as a result of incomplete combustion. The smoke
56. 56
produced depends on the amount of air available and the
quality of wood or sawdust. Soft woods produce a lot of smoke,
which may lead to blacking of the finished products. Wood
smoke is a mixture of complex chemical product gases, vapor
and volatile substances. The volatile substances are absorbed
on the wet surfaces of fish during the smoking and produce the
characteristic aroma (FAO/UN, 1971a).
As it is frequently seen in fish markets, properly smoked
fish products are dark brown in colour and are mostly near
perfectly dried. This ensures that the shelf life is prolonged and
the products get the consumer in relatively good stat (FAO,
1971).
57. 57
2.2.6.4 Salting
There are four standard methods for salting fish. These
are brine. Dry, kench and pickle salting methods. In brine
salting, the fish are immersed in a solution of salt in water.
Where granular salt is rubbed into the surfaces of fish, the
process is referred to as dry salting. Granular salt is also used
in kench salting. In this process, the salt is rubbed into the
surface of split fish and the fish are stored with salt placed
between each layer of fish. The liquid formed to drain off the
fish, which will eventually become covered with the liquid. The
liquid is referred to as pickle. In pickle salting, the fish are
packed in water to flat containers with salt between each layer
58. 58
of fish. If the pickle formed does not cover the fish within 4h,
saturated brine id added to the fish so that, it become
immersed by the pickle. Otherwise, the fish may spoil (FAO,
1971).
In brine salting, a saturated brine solution is used. Brine is
prepared by dissolving 270-360g of salt in one liter of water.
Fish are then completely immersed in the solution. Due to
uptake salt, the concentration in brine drops as consequence of
water exuding from the fish. Fish may be stored occasionally to
enhance the uptake of salt, the concentration in brine drops as
consequences of water exuding from the fish. Fish may be
59. 59
stored occasionally to enhance the uptake of salt. The latter
may be eliminated if the brine is much (FAO, 1981).
2.2.6.5 Fermentation
Claria species are also frequently, fermented. They are
gutted cleaned of the gills, salted and are placed in concrete
tanks. Organic acid is added and the fish remain in the brine for
about four months. The end product keeps from up to one
year, where fermentation is allowed to continue for a very long
time, sauces are produced. Sances are liquids containing
mixtures of amino acids and protein degradation products.
60. 60
They have very high salt content and may provide a good
flavor. In preparing fish sauces, the fish is submerged in brine
for up to 18 months. The ripe sauce colour ranges from yellow
to dark brown. The aroma and flavor are characteristic and
determine the grade. Sauces are stable and may be kept for
extremely long time (Oyelese, 2006).
The successful preservation of fish b biological
fermentation method is depended on the production of lactic
acid. Lactic acid bacteria ferment the sugars present to organic
acid resulting in the lowering of pH the low pH inhibits growth of
pathogen or faddisms and putrefactive organisms. Since fish
contains only small amount of fermentable carbohydrates,
61. 61
mixtures of malt, corn or tapioca should be added (Tawari,
2006).
CHAPTER THREE
3.0 MATERIALS AND METHODS
3.1 Plant Sample Collection
62. 62
Fresh Moringa oleifera leaves were collected from
the Unical Farm, University of Calabar, Calabar and
identified by expert in department of Botany, University
of Calabar. The collected samples were stored all room
temperature in a closed environment.
3.2 Preparation of Marinade
Fresh Moringa leaves were air dried for 4days at
ambient temperature (37±20
c) and ground into power
using a food blender. Moringa Oleifera Marinade (MOM)
was prepared by adding separately specific quantity
(10g, 20g or 30g) of Moringa oleifera leaf powder to
1000ml of water to form 1, 2 or 3% Moringa oleifera
Marinade respectively. 50g of salt and 2g of BHA was
added separately to 1000ml of water to form 5% Brine
and 0.2% BHA solution. No additive was added to the
control treatment.
63. 63
3.3 Fish Preparation
Thirty Catfish samples were purchased from Watt
Market in Calabar, Cross River State. The processing
and smoking of the fishes were carried out at the
University of Calabar fish farm. The average weight of
the fishes was 230±8g while the total weight was 20kg.
The fishes were gutted using a sharp knife by cutting
laterally from the end of the gill cover through the belly
portion to the anus. Thereafter, they were thoroughly
washed and rinsed. The total weight of the fish after
gutting was 18.5kg with the average weight of 220±6g.
3.4 Fish Treatment
The fishes were randomly assigned to six
experimental groups of treatments: control,
1%Moringa oleifera Marinade, 2%Moringa oleifera
64. 64
Marinade, 3%Moringa oleifera Marinade, 5% salt and
0.2%BHA. Each treatment was replicated with 5
fishes/replicate. The fishes were soaked in the
marinade for 2hours. Thereafter, the fishes were set in
the smoking kiln consisting of five-twin tiers and
subjected to hot smoking for 12 hours with charcoal as
the heat source. The tiers were interchanged every
3hours to ensure uniform heat distribution and drying.
The smoke-dried fishes were placed in air-free netted
boxed to prevent flies infestation and stored at room
temperature (37±20
c) for 3weeks.
3.5 Sensory analysis
A thirty member panel conducted the sensory
analysis. The taste panelist (assessors) were drawn
from the Watt Market fish sellers and fish buyers. They
65. 65
were instructed on the parameters to adjudge using a
9-point hedonic scale. The hedonic scale is delineated
as follows: 9 = Like extremely, 8 = Like very much, 7
= Like moderately, 6 = Like Slightly, 5 = Neither like
nor dislike, 4 = Dislike very much, 1 = Dislike
extremely. Water was provided for the panelists to
rinse their mouth after each bite to eliminate the taste
of the previous fish sample.
3.6 Statistical Analysis / Data Evaluation
The assessors scores were used to analyze the shelf-life
of the fish by considering the storage period from which the
fish become vulnerable to microbial infestation which can be
determine by Moisture content, change in taste, colour and
flavor (odour) of the each of the six groups of the fish used for
66. 66
the research. All the data obtained were subjected to analysis
of variance (ANOVA) to determine the levels of significance.
(Steel and Torrie, 1990).
67. 67
CHAPTER FOUR
4.0 RESULTS
The percentage change in moisture content of the fish
samples at different days interval after treatment with Moringa
oleifera Marinade fractions, salt and BHA in (W/V) solutions.
The 3% Moringa oleifera Marinade showed the least
mean of 3.0±0.28 moisture content at the end of 21 days
(3weeks) of treatment compared to the control having mean
value of 17.60±0.46 moisture content. The BHA and control
treated samples have the highest mean value of 15.40±0.22
and 17.60±0.46 moisture content respectively.
68. 68
The result shows that the major dehydrating
component(s) of the plant is concentration dependent. All the
Moringa oleifera Marinade treated samples shows significant
dehydrating property having significant difference (P<0.01)
when compared to the control.
The taste was observed to show no significant difference
(P>0.01) among the Moringa oleifera Marinade treated
samples. 3% Moringa oleifera Marinade and 2% Moringa
oleifera Marinade were scored higher as 3% Moringa oleifera
Marinade shows highest mean value of 7.4±0.83, followed by
2% Moringa oleifera Marinade with mean value of 7.0±0.40
compared to the control and BHA of mean value 3.6±0.46 and
69. 69
3.6±0.46. The control and BHA treated sample shows no
significant difference (p>0.01).
The result of the flavour shows no significant difference
(p>0.01) among the control and the Moringa oleifera treated
samples at the 21 Day (3weeks) of storage. BHA shows the
least mean value of 3.2±0.54 and also the least in colour
analysis with the mean least 3.2±0.54 which the Moringa
oleifera Marinade and the control shows no significant
difference (P>0.01) in colour changes.
Table 4a: Showing the effect of treatment on the
Moisture content
Treatment Storage period in days
Day 3 Day 6 Day 9 Day 12 Day 15 Day 18 Day 21
70. 70
Control 22.4±0.61 14.6±0.46 12.4±0.43 16.0±0.63 23.2±0.72 16.4±0.38 17.6±0.46
1% MOM 10.0±0.63 9.6±0.36 7.8±0.54 14.2±0.78 9.2±0.38 9.2±0.38 6.6±0.36
2% MOM 10.4±0.46 8.2±0.54 8.8±0.54 12.0±0.63 8.2±0.38 8.2±0.38 5.4±0.36
3% MOM 8.2±0.54 5.4±0.36 4.8±0.54 6.2±0.38 4.2±0.44 4.2±0.44 3.0±0.28
5% MOM 9.8±0.38 6.4±0.22 6.2±0.30 10.0±0.40 6.0±0.28 6.0±0.28 8.2±0.62
0.2%BHA 15.0±0.57 13.6±0.54 13.8±0.38 16.0±0.28 15.2±0.38 15.2±0.38 15.4±0.22
Table 4b: Showing the effect of treatment on the colour
change
Treatment Storage period in days
Day 3 Day 6 Day 9 Day 12 Day 15 Day 18 Day 21
Control 7.8±0.30 8.0±0.49 7.4±0.49 5.4±0.48 7.2±0.29 6.2±0.38 6.8±0.78
1% MOM 7.4±0.22 8.4±0.22 7.8±0.30 7.8±0.30 7.8±0.38 7.0±0.28 6.4±0.31
2% MOM 7.0±0.28 6.0±0.63 6.6±0.22 7.0±0.49 7.0±0.57 6.8±0.38 6.6±0.22
3% MOM 7.0±0.63 7.4±0.67 7.8±0.30 6.4±0.78 8.0±0.49 7.4±0.22 6.0±0.28
5% MOM 6.2±0.56 6.2±0.30 4.6±0.61 7.2±0.62 5.6±0.19 6.6±0.22 5.4±0.22
0.2% BHA 8.0±0.28 7.2±0.30 5.2±0.30 6.2±0.38 8.0±0.28 6.6±0.67 3.2±0.54
Table 4c: Showing the effect of the treatment on the
flavour change
Treatmen Storage period in Days
71. 71
t Day 3 Day 6 Day 9 Day 12 Day 15 Day 18 Day 21
Control 7.6±0.22 7.6±0.36 7.4±0.22 4.8±0.30 6.6±0.22 7.2±0.30 7.2±0.38
1% MOM 7.0±0.28 6.6±0.22 7.8±0.58 7.2±0.38 7.0±0.40 7.0±0.28 6.4±0.36
2% MOM 7.6±0.22 6.8±0.31 6.4±0.22 6.2±0.30 6.8±0.31 7.0±0.28 6.4±0.31
3% MOM 7.0±0.28 7.4±0.22 7.8±0.30 6.8±0.49 7.2±0.38 7.0±0.63 6.8±0.54
5% MOM 7.0±0.28 5.8±0.38 4.6±0.22 5.8±0.38 5.6±0.22 6.6±0.23 5.4±0.22
0.2% BHA 7.4±0.36 7.4±0.36 5.2±0.35 5.2±0.38 7.8±0.30 7.0±0.28 3.2±0.54
Table 4d: Showing the effect of treatment on the taste
change
Treatment Storage period in Days
Day 3 Day 6 Day 9 Day 12 Day 15 Day 18 Day 21
Control 8.2±0.20 7.8±0.30 7.8±0.38 5.2±0.20 6.4±0.30 5.0±0.40 3.6±0.46
1% MOM 8.0±0.28 8.4±0.22 7.8±0.30 7.0±0.28 6.8±0.38 6.8±0.54 6.0±0.28
2% MOM 7.6±0.46 8.0±0.28 6.2±0.38 6.4±0.46 6.4±0.61 7.2±0.49 7.0±0.40
3% MOM 7.4±0.22 7.0±0.28 8.0±0.28 7.8±0.54 8.2±0.54 7.0±0.28 7.4±0.83
5% MOM 7.2±0.15 6.4±0.36 4.8±0.38 6.2±0.62 6.0±0.28 6.8±0.51 6.0±0.45
0.2%BH
A
7.6±0.46 7.6±0.46 5.2±0.62 5.6±0.22 7.6±0.22 7.0±0.40 3.6±0.46
72. 72
CHAPTER FIVE
5.0 DISCUSSION, CONCLUSION AND
RECOMMENDATION
5.1 Discussion
During the analysis, thirty assessors were used to carry
out the test base on each assessor score and the mean and
standard error of mean was calculated and use for as result.
The percentage change in moisture content of the fish
mode at different day interval after the treatment with Moringa
oleifera leave fractions, salts solution and butyrate hydroxyl
anisole (BHA) is table 4a. The general decrease in moisture
73. 73
content was observed from the first week to the 3 weeks of
experiment. Fish samples treated with 3% Moringa oleifera
marinade (MOM) was observed to have the lowest moisture
content which was significantly different from that of other
treatments throughout the storage period.
This moisture content decrease was followed by 2%
Moringa oleifera Marinade, 1% Moringa oleifera Marinade and
salt treated samples. There was no significant difference
(p>001) among fish samples treated with 1% Moringa oleifera
Marinade, 2% Moringa oleifera Marinade and salt on the third
day of storage. The control samples had the moisture content
of the control and BHA treated samples exceeded the
74. 74
benchmark of 10% (Daranola et al., 2007) and 12%
(Seviensdothir, 1998) when microbial spoilage might occur. The
moisture content of all Moringa oleifera treated samples in the
day 21 was significantly lowered than what was observed in the
third day of storage. The moisture content of BHA and control
samples followed the same trend. Although, BHA is a potent
antioxidant (Adeyemi et al, 20011), it impacted negatively on
the moisture content of chorkor smoked fish sample.
The ability of Moringa oleifera marinade to reduce the
moisture content of the fish sample could result from myriad
chemical compounds (that have high dehydrating properties)
e.g calcium salts (Grubben and Denton, 2004), prevent
75. 75
in Moringag oleifera leaves from which the marinade was
prepared. Salt decrease the water activity of podis; the
mechanism involves transporting salt into food structures and is
achieve by various physical and chemical factors such as
diffusion , osmosis and a series of complicated chemical and
biochemical processes (Turan et al., 2007). A high
dehydrating, prospecting exhibited by Moringa oleifera
and salt is beneficial in that microbial deterioration is inhibited
thus enhancing the shelf-life of chorkor smoke fish. in addition
both salt and marinade are cheap and affordable.
There was no significant difference (p>001) between the
control and the Moringa oleifera treated samples with respect to
76. 76
colour observed by the assessors (table 4b). A visual
observation of the fishes showed that all samples had dark
brownish colour. This colour was due to smoke and ashes
deposited on the fishes during smoke-drying process. Moringa
oleifera treated samples had dark greenish patches on them
intensity of the greenish patches was concentration dependent
with 1% Moringa oleifera Marinade having the least.
There was no significant difference (p>0.01) between the
control and the moringa oleifera treated samples at the day 21
of storage in Table 4c. BHA showed a high level of decrease in
flavor scored by the assessors at the end of the day 21,
followed by salt treated group. There were fluctuations, in the
77. 77
flavor scored observed for all treatment. The scores for all
moringa treated samples in the day 21 do not differ from that of
the day 3.
There was no significant difference (p>0.01) observed
between the Moringa oleifera treated samples in their taste. But
a significant difference ( p<0.01) was observed between
sample. This 2% and 30% Moringa oleifera Marinade treated
sample were scored significantly higher than other treated
sample, followed by 1%Moringa oleifera Marinade and 5%
Brine which were observed to show no significant difference
( p>0.01) at the end of the experiment.
78. 78
Progressed reduction in taste, color and flavor scares
storage could be attributed to the activity of spoilage agents.
Quality loss during storage at both ambient temperature and
chilling was revealed in the results of research of Oyster
(Liobredaa et al, 1986) and shrimps (Reithy and Dela-cruz,
1986). To make choice of fish, physical assessment methods is
easily available to an intending fish buyer.
5.2 Conclusion
All samples treaded with Moringa Oleifera marinade
showed reduced moisture content. The 3% Moringa oleifera
79. 79
Marinade treated sample showed a significant (p<0.01)
reduced moisture content, compared to other samples; and this
was followed by 2% Moringa oleifera Marinade and 1% MOM
orderly.
Control samples had the highest moisture content, which
was closely followed by BAA samples. The dehydrating
properly of Moringa oleigera is depends on concentration. It is
well known that increase in moisture content of samples
increases the vulnerability of the samples to microbial
infestation resulting in spoilage.
80. 80
Moringa oleidfera marinade could be used to maintain the
organoleptic property and improve shelf-life of chorkor smoked
African catfish for as long as the period of 3 weeks.
5.3 Recommendation
This work showcase the effect of Moringa oleifera
marinade on a chorkor smoked fish, with regard to the shelf-life
and the organoleptic properties. I will therefore recommend
further research on the specific phytochamical(s) or mineral(s)
81. 81
content of the plant that gives it such remarkable dehydrating
and preservative properties.
82. 82
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106. 106
Showing moisture content of the sample
Figure 3. Percentage Moisture Content of the different
experimental groups. Values are expressed
as percentage mean±SEM, n=5
*significantly different from NC at p<0.01;
108. 108
Figure 4. Percentage Moisture Content of the different
experimental groups. Values are expressed
as percentage mean±SEM, n=5
*significantly different from NC at p<0.01;
Showing colour change
109. 109
Figure 5. Colour change of the different
experimental groups. Values are expressed
as mean±SEM, n=5
*significantly different from NC at p<0.01;
110. 110
Showing colour change
Figure 6. Showing Colour Change of the different
experimental groups. Values are expressed
as mean±SEM, n=5
111. 111
*significantly different from NC at p<0.01;
Showing Flavour change
Figure 7 . Showing Flavour Change of the different
experimental groups. Values are expressed
113. 113
Figure 8.Showing Flavour Change of the different
experimental groups. Values are expressed
as mean±SEM, n=5
*significantly different from NC at p<0.01;
Showing taste change
114. 114
Figure 9. Showing Taste Change of the different
experimental groups. Values are expressed
as mean±SEM, n=5
*significantly different from NC at p<0.01;
115. 115
Showing taste change
Figure 10. Showing Taste Change of the different
experimental groups. Values are expressed
as mean±SEM, n=5
*significantly different from NC at p<0.01;
