EVALUATION MORPHOLOGY, GROWTH
PERFORMANCE AND SENSORY ATTRIBUTES OF FOUR RED HYBRID TILAPIA (OREOCHROMIS SP.)
STRAINS.
MOHD ZHARIF BIN RAMLI
MASTER OF SCIENCE
2017
Evaluation of Morphology, Growth Performance and Sensory Attributes of Four Red Hybrid Tilapia
(Oreochromis Sp.) Strains.
by
MOHD ZHARIF BIN RAMLI
A thesis submitted in fulfillment of the requirements for the degree of Master of Science
Faculty of Agro Based Industry
UNIVERSITI MALAYSIA KELANTAN
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THESIS DECLARATION
I hereby certify that the work embodied in this thesis is the result of the original research and has not been submitted for a higher degree to any other University or Institution
OPEN ACCESS I agree that my thesis is to be made immediately available as hardcopy or on-line access (full text) EMBARGOES I agree that my thesis is to be made available as
hardcopy or on-line (full text) for a period approved by the Post Graduate Committee
Dated from until
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RESTRICTED (Contains restricted information as specified by the organization where the research was done)*
I acknowledge that Universiti Malaysia Kelantan reserves the right as follows.
1. The thesis is the property of Universiti Malaysia Kelantan.
2. The library of Universiti Malaysia Kelantan has the right to make copies for the purpose of research only.
3. The library has the right to make copies of the thesis for academic exchange.
SIGNATURE SIGNATURE OF SUPERVISOR
IC/PASSPORT NUMBER NAME OF SUPERVISOR
Date: Date:
ACKNOWLEDGEMENTS
In The Name of Allah, The Most Beneficent, The Most Merciful
I would like to extend my greatest gratitude to Allah in such His blessing for me to explore new knowledge and broadening the perspective in the research. Firstly, my sincere thanks goes to my main supervisor Assoc. Prof. Dr. Lee Seong Wei and co- supervisor Prof. Dr. Wan Zahari Bin Mohamed. Not to forget, my former co- supervisor Dr. Soon Jan Mei as well. My appreciation for precious support, guidance, advice and immense knowledge throughout the study and thesis completion. I certainly honored to know them since my first glance as researcher. Special thanks to Universiti Malaysia Kelantan and the Ministry of Higher Education of Malaysia for funding recent study through the Niche Research Grant Scheme (NRGS) (R/NRGS/A07.00/00387A/006/2014/000152) and MyBrain. Furthermost, I would like to convey appreciations to my parents, Ramli bin Mat and Morni bt Arshad for all- inclusive and absolute support. To my sisters and brothers for supporting me spiritually throughout this study and my life as well. Last but not least, I should say thank you to my buddies Zack, Arep, Zaty, Zee and Martina for their dedication of time, energy and expertise since day one. Not to forget to remark, to all my friends and staffs for their assistance all over the research and dissertation completion. Thank You (Zharif, R., 2017).
May Allah reward you for the good
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TABLE OF CONTENTS
PAGE
THESIS DECLARATION i
ACKNOWLEDGEMENT ii
TABLE OF CONTENTS iii
LIST OF TABLES vii
LIST OF FIGURES viii
LIST OF ABBREVIATIONS ix
LIST OF SYMBOLS x
LIST OF EQUATIONS xi
ABSTRAK xii
ABSTRACT xiii
CHAPTER 1 INTRODUCTION
1.1 General introduction of tilapia 1
1.1.1 Taxonomy and general morphology of tilapia 6
1.2 Problem statements 9
1.3 Significance of study 9
1.4 Objectives 10
1.5 Scope of study 10
CHAPTER 2 LITERATURE REVIEW
2.1 Breeding techniques, nutrition and ecological parameters 11
2.1.1Broodstock management and fry production 13
2.2 Phenotype (morphometric characteristics) characterisation 16 2.3 Evaluation of Growth Performances, Feed Conversion Ratio (FCR),
Specific Growth Rate (SGR) and Survival Rate (SR). 24
2.4 Sensory analysis 29
CHAPTER 3 METHODOLOGY
3.1 Experimental design in producing four strains of red hybrid tilapia. 34
3.1.1 Broodstocks management 36
3.1.2 Broodstock nutrition 37
3.1.3 Collection of fries 38
3.2 Growth performance evaluation 38
3.2.1 Introduction of fry into hapas 39
3.2.2 Fry nutrition 39
3.2.3 Weight measurement 40
3.2.4 Growth performance analysis 40
3.2.5 Statistical analysis 42
3.3 Morphometric characteristics measurement 43
3.4 Sensory analysis 47
3.4.1 Sample preparation 47
3.4.2 Sensory test conditions 48
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3.4.3 Characteristics of the evaluators 49
3.4.4 Analysis of sensory attributes 49
3.4.5 Statistical analysis 49
CHAPTER 4 RESULTS
4.1 Characterisation of red hybrid tilapia morphology 50 4.1.1 A summary of morphometric characteristics measured in a ratio to
total length (TL) and logarithm for two broodstocks of red hybrid tilapia.
50 4.1.2 A summary of morphometric characteristics measured in a ratio to
total length (TL) for four strains of red hybrid tilapia 55
4.1.3 Morphology characteristics distance 59
4.2 Growth performance of four strains red hybrid tilapia. 63
4.2.1 Water parameters 63
4.2.2 Growth performances 64
4.3 Sensory analysis 66
4.3.1 Characterisation of surveyed participants 66
4.3.2 Sensory analysis attributes on four strains of redtilapia. 70
4.3.4 Ranking test 73
CHAPTER 5 DISCUSSION 74
CHAPTER 6 CONCLUSION 91
REFERENCES 94
APPENDIX A 105
APPENDIX B 107
APPENDIX C 108
APPENDIX D 109
APPENDIX E 115
LIST OF PUBLICATIONS 116
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LIST OF TABLES
NO. PAGE
1.1 Taxonomy of Oreochromis niloticus and Oreochromis mossambicus 7 2.1 The summary of phenotypic measures in characterising tilapia stocks. 23
2.2 The summary of growth performance of tilapia 28
2.3 The summary of sensory analysis on fish attributes. 33
3.1 Broodstock ratio with different combination 36
3.2 Changes in weight and CP (%) according to life stages (fry to adult). 40 3.3 Quantitative phenotype traits based on morphometric characteristics
for differentiation analysis among the red hybrid tilapia
46 4.1 Mean ± S.D of quantitative phenotype traits based on morphometric
characteristics ratio for differentiation analysis, among different four strains of red hybrid tilapia.
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4.2 The morphological distance between different strains based on morphological ratio.
62 4.3 Average values with S.D of water temperature, pH, and dissolved
oxygen (O2D) and salinity during the experiment period.
63 4.4 Growth performance and survival rate of four strains of red hybrid
tilapia.
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4.5 Sensory rating differences obtained from four strains red hybrid tilapia.
70 4.6 Summary of correlation coefficients between sensory attributes for
four red hybrid tilapia strains.
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LIST OF FIGURES
NO. PAGE
1.1 General morphology of red hybrid tilapia (Oreochromis sp.) 8 3.1 Morphometric characteristics of red hybrid tilapia broodstocks. 35
3.2 Experimental hapas in the earthen pond 37
3.3 Four strains of red hybridtilapia 44
3.4 Morphometric characteristics indices 45
4.1 Red hybrid tilapia O.niloticus and O.mossambicus. 52
4.2 Mean ratio of morphometric characteristics of male broodstocks 53 4.3 Mean ratio of morphometric characteristics of female broodstocks 53 4.4 Mean log of morphometric characteristics of male broodstocks 54 4.5 Mean log of morphometric characteristics of femalebroodstocks 54 4.6 Morphometric distance relationship taxa inferred using the UPGMA
method.
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4.7 Morphometric distance taxa relationship inferred using the average linkage (between groups) in hierarchical cluster analysis method.
61 4.8 Percentage (%) of evaluators’ gender participated in sensoryanalysis. 67 4.9 Percentage (%) of evaluators’ age participated in sensory analysis. 68 4.10 Percentage (%) of evaluators’ races participate in sensoryanalysis 69 4.11 Preference ranking of four strains of red hybrid tilapia ranked by
evaluators in sensory analysis.
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LIST OF ABBREVIATIONS
RGR Relative Growth Rate
SGR Specific Growth Rate
FCR Feed Conversion Ratio
SR Survival Rates
FM Fish Meal
O2D Dissolved Oxygen
CP Crude Protein
GIFT Genetic Improved Farmed Tilapia
LIST OF SYMBOLS
% Percentage
± Plus Minus
oC Degree Celcius
mg/l Milligram per liter
g gram
kg kilogram
MT Metric Tones
ppt Part per thousand
mm millimeter
m2 Meter square
cm centimeter
m meter
♀ Female
♂ Male
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LIST OF EQUATIONS
PAGE
Relative Growth Rate (RGR) 41
Feed Conversion Ratio (FCR) 41
Specific Growth Rate (SGR) 41
Survival rate (SR) 41
Penilaian Morfologi, Prestasi Pertumbuhan dan Ciri-ciri Sensori pada Empat Jenis Tilapia Merah Hibrid (Oreochromis Sp.)
ABSTRAK
Penurunan hasil pengeluaran tilapia disebabkan oleh kemerosotan ciri-ciri baik pada ikan tilapia tersebut. Kajian terkini menyelidik tilapia merah hibrid dengan menekankan ciri-ciri fizikal, tumbesaran dan rasa yang berpotensi untuk dipertingkatkan. Empat baka tilapia merah hibrid dihasilkan melalui empat kombinasi yang berbeza; (i) ♂O.niloticus x ♀O.niloticus (A), (ii) ♂O.mossambicus x
♀O.mossambicus (B), (iii) ♂O.niloticus x ♀O.mossambicus (C) dan (iv)
♂O.mossambicus x ♀O. niloticus (D). Anak ikan dipelihara dalam 12 hapas (3.0 m x 2.0 m x 1.2 m) di dalam kolam tanah dengan diberi makan makanan komersial (35- 40% Protein Kasar) 5% daripada berat badan dengan kepadatan 10 individu/m3 selama 70 hari. Dua puluh tiga ciri-ciri morfometri dan 6 parameter prestasi tumbesaran dinilai secara serentak. Manakala, analisis sensori pula dijalankan selepas 4 bulan ikan dibesarkan di dalam kolam tanah. Data morfometri dianalisis dalam nisbah allometri diikuti dengan analisis kluster hieraki. Ciri-ciri morfometri dan prestasi tumbesaran dinilai melalui analisis varians (ANOVA). Keputusan kajian menunjukkan 9 ciri-ciri seperti lebar badan ( Bw1 ) , jarak antara duri akhir sirip dorsal dengan duri pertama sirip anal ( D1da ), jarak sirip kaudal dengan sirip anal ( DcaL ), ketinggian badan ( BH) , panjang pra- pektoral (PCL ), panjang pra-pelvik (PPL ), panjang pra -anal (PAL ), panjang pra-orbit (POL) , panjang sirip pelvik ( PlvL ) menunjukkan perbezaan yang signifikan (p<0.05) antara baka. Analisis kluster hieraki memperincikan baka ini kepada 2 kumpulan dan 1 sub-kumpulan yang disimpulkan sebagai perbezaan pada baka. Berat mula (Wi), Berat akhir (Wf), Perbezaan Kadar Pertumbuhan (RGR), Kadar Pertukaran Makanan (FCR), Kadar Pertumbuhan Spesifik (SGR) and Kadar Survival (SR) dinilai. Paraneter ini tidak berbeza secara statistik (p<0.05) di antara baka. Walau bagaimanapun, baka A mempunyai skor yang paling tinggi iaitu Wf (19.63±0.69), RGR (2357.06±66.78), FCR (1.33±0.02) dan SGR (4.57±0.03) secara perangkaan. Min prestasi tumbesaran paling rendah direkodkan pada baka B iaitu Wf (17.772±0.98), RGR (2130.88±92.14), FCR (1.50±0.02) dan SGR (4.43±0.06).
Manakala, prestasi tumbesaran baka C dan D pula didapati pada nilai min purata. SR direkodkan tinggi (>90%) pada semua baka dengan tiada perbezaan signifikan (p<0.05). Data analisis sensori yang tidak parametrik dinilai menggunakan ujian Kruskal-Wallis. Secara umumnya, tiada perbezaan signifikan (p>0.05) antara ciri-ciri (rasa, bau dan jusnya) kecuali pada tekstur.Hasil kajian ini menunjukkan ciri-ciri unik yang membezakan baka-baka dan kombinasi pembiakbakaan efektif dalam menghasilkan baka dengan ciri-ciri yang baik. Penglibatan ♀ O.niloticus ialah elemen utama dalam peningkatan ciri-ciri pada tilapia hibrid merah.
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Evaluation of Morphology, Growth Performance and Sensory Attributes of Four Red Hybrid Tilapia (Oreochromis Sp.) Strains.
ABSTRACT
Deterioration of good characteristics in tilapia stocks causes the decline in its production yield. Current research investigated red hybrid tilapia strains by emphasising on physical, growth and taste characteristics which have the potential to be improved. Four red hybrid tilapia strains were produced by four different combinations; (i) ♂O.niloticus x ♀O.niloticus (A), (ii) ♂O.mossambicus x
♀O.mossambicus (B), (iii) ♂O.niloticus x ♀O.mossambicus (C) and (iv)
♂O.mossambicus x ♀O. niloticus (D). Fries were reared in 12 hapas (3.0 m x 2.0 m x 1.2 m) in earthen pond, fed 5% of their body weight with commercial feed (35- 40%Crude Protein) at stocking density of 10 individuals/m3 for 70 days. Twenty-three morphometric characteristics and 6 parameters of growth performance were evaluated concurrently. Meanwhile, sensory analysis of the fish was carried out after 4 months of being raised in the earthen pond. The morphometric data were analysed in allometric ratio followed by hierarchical cluster analysis. Morphometric and growth performance characteristics were analysed by analysis of variance (ANOVA).The results showed 9 characteristics which are body width (Bw1), distance from end of dorsal ray to 1st anal fin (D1daL), distance from caudal to anal fin (DcaL), body height (BH), pre-pectoral length (PCL), pre-pelvic length (PPL), pre-anal length (PAL), pre-orbital length (POL), pelvic fin length (PlvL) were found to be significantly different (p<0.05) between these strains. Hierarchical cluster analysis characterised these strains into 2 groups and 1 sub-group which inferred the variation between strains. Initial weight (Wi), Final weight (Wf), Relative Growth Rate (RGR), Feed Conversion Ratio (FCR), Specific Growth Rate (SGR) and Survival Rate (SR) were calculated. No statistical difference (p<0.05) between evaluated strains were found in these parameters.
However, strain A had the highest score in Wf (19.63±0.69), RGR (2357.06±66.78), FCR (1.33±0.02) and SGR (4.57±0.03) in numerical data. Lowest mean values of growth performances were recorded on strain B which was Wf (17.77±0.98), RGR (2130.88±92.14), FCR (1.50±0.02) and SGR (4.43±0.06). Meanwhile, the growth performance of strains C and D were found to be at average mean value. SR was highly scored (>90%) in all strains with no statistical difference (p<0.05). Non- parametric data of sensory analysis were analysed by Kruskal-Wallis test. Generally, there is no significant difference (p>0.05) between attributes (taste, odour and juiciness) except for texture. These findings showed that unique characteristics distinguished between strains and combination of breeding effectively produced strains with good characteristics. Utilisation of ♀ O.niloticus is the key element for characteristics improvements in red hybrid tilapia strains.
CHAPTER 1
INTRODUCTION
1.1 General introduction of tilapia.
In the aquaculture industry, several species of tilapia were introduced for the purpose of human consumption. The Nile tilapia (Oreochromis niloticus L.) from the Cichlidae family is presently the major species introduced into aquaculture for commercialisation, consumption and research purposes. It is easy to breed and managed in various types of aquaculture systems, fast growers, effective feed converter, resistant to certain diseases and endures the changes in water parameters (Salah, 2008) It was reported that, Malaysia produced 44,099.24 tons of tilapia in 2013; 80% of which were red tilapia (Mazuki, 2015).
In the mid-80’s, the farming of this high value aquaculture species started in Malaysia, yielding the highest production of red hybrid tilapia in 2012 (52,295.30 MT) (Mazuki, 2015). This statistic was recorded due to the high demand of this tilapia strain compared to Nile tilapia and Genetically Improved Farmed Tilapia (GIFT) developed by World Fish, Malaysia (Olievera et al., 2014). In fact, the wholesale value of aquaculture production of freshwater culture showed that red tilapia and black tilapia were sold at RM 246,368.46 /MT and RM 33,122.54
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Red tilapia remains highly favoured in the market due to its hardy characteristics that farmer finds rewarding. In addition, attractive attributes like its red colour and ‘clean’ appearance made this strain highly valued (Popma &
Lovshin, 1995; Allaman et al., 2013). In 1960’s, Taiwanese red hybrid tilapia was produced in Taiwan resulted by crossing between a normal male O. niloticus with mutant reddish-orange female O. mossambicus (Popma & Masser, 1999).
Meanwhile in Florida in the 1970’s, red tilapia strain was produced resulted by crossing a red-gold O. mossambicus with normal coloured female O. hornorum (Popma & Masser, 1999). Crossing a wild Blue tilapia (O. aureus) with mutant pink Nile tilapia produced a red tilapia strain in Israel (Popma & Masser, 1999).
These original red hybrid tilapia strains were crossed with other Oreochromis species and distributed around the world. In fact, 70 domesticated species are commercially important, thus cultured around the world (Peterman, 2011). Red hybrid tilapia strains were widely farmed In South East Asia, O.mossambicus in Thailand and O.niloticus and Oreochromis sp. in Malaysia (Pongthana et al., 2010).
History has revealed how the red hybrid tilapia was established. Breeding programme has led to a significant implication in red hybrid tilapia production and improvement. Previously, several works were reported in improving the red hybrid tilapia performance in an aquaculture system aimed to increase salinity tolerance, growth rates, disease resistance and product quality (Hernández et al., 2001; Pongthana et al., 2010; Thodesen et al., 2013). However, red hybrid tilapia
experienced introgressive hybridization due to inappropriate management and some breeding programmes involved were not reported. As a result, red hybrid tilapia was changed in terms of its genetics and appearances, which contributed to the declining quality of economic traits (Karuppannan et al., 2013).
Breeding goal is significant in designing a breeding programme and its management; particularly in tilapia production, and it is a dynamic activity.
Previous studies were carried out aimed for product quality improvement (Hernández et al., 2001), production cost reduction (Workagegn & Gjoen, 2012), external colour traits (Thodesen et al., 2013) and etc. In order to achieve these goals, aquaculturist device a breeding programme involving several species and improved strains of tilapia. Hybridization appears to be a promising option breeding programme in tilapia; regardless interspecific or intraspecific, the finding was remarkable. More often than not, breeding goals were emphasised on the economical traits possessed by the targeted fish. For example, the economic traits like growth rate, survival rate and the appearance of food fish. These traits changes as time passes since the viability of the food fish is determined by the consumers or market demand. For instance, GIFT is a successful strain that produced through a breeding programme which have a higher growth rate and survival (Gjerdrem, 2012). In fact, the success of the breeding programme is dependent upon the basic principles of hybridization; where species or strains chosen must contain variation (Gjerdrem & Baranski, 2009).
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Gjerdrem & Baranski (2009) highlighted that variation is an essential condition before a breeding programme can be initiated. The selected traits must vary between individuals since portion of the variation is contributed by genetic differences. These traits were anticipated to be inherited by progeny. Variation in the population can be characterised by their phenotypes, growth performances and quality of sensory attributes (Gjerdrem & Baranski, 2009). Variability of traits provide information about the species which can possibly be utilised in a breeding programme.
Variations in phenotypes segregate the species and indicate the economically desired morphology (Škrijelj et al., 2011). Morphometric characteristics apparently was discovered as a method to identify fish population, which is of great interest in tilapia aimed for population characterisation, their growth performance and useful for taxonomic status (Amon et al., 2013; Melo et al., 2013; Oliveira et al., 2014). This method is widely practiced in fisheries and aquaculture studies since it can directly identify the species without killing the animals (Kosai et al., 2014). However, the analysis of morphology indicates the variation in physical appearance traits but not directly the species. Therefore, species characterisation and genetic marker information are complementing one another in making the analysis a reliable one. In spite of that, the recent investigation emphasises on the biometric study, which also concerns the verification of the speciesinvolved.
In addition, the variation in growth performance can also be influenced by genetics besides food quality and environmental factors. Growth performance variation between strains could be inferred as a result of genotype differences (Workgagegn & Gjoen, 2012). This work is vital in selecting the best strains for an effective breeding programme which possess traits that can be incorporated into the breeding goals. In fact, these economic traits were reported to have reduced production cost, efficiently utilised the feed, and improved the product quality (Workgagegn & Gjoen, 2012).
Each aquaculture product has a distinct consumer preference. Market demands quickly change due to the fluctuation of consumer preference and eating behaviour which pave the industry striving to satisfy their demands (Silva et al., 2015). In general, aquaculture products were designed to achieve desirable sensory attributes of the species in terms of tenderness, flesh colour and flavour (Kilcast, 2000). Aquaculturist satisfied the consumer demand through breeding programme, utilisation of optimal customised feeds, culture technologies and proper management practices (Hernández et al., 2001; Hernández et al., 2007). Sensory attributes can possibly be improved through these approaches, though it is challenging to create a quality guideline due to the fluctuation in consumer demands. For instance, the high performance GIFT had failed to compete with the production of red hybrid tilapia in Malaysia as shown in the previous statistics.
This situation occurred due to consumers’ preference for the red hybrid tilapia,
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susceptible to consumer preferences hence require modifications in the breeding goal, trait assessment and other farming practices over time (Gjerdrem & Baranski, 2009).
1.1.1 Taxonomy and general morphology of tilapia
‘Tilapia’ is a common name in several countries around the world.
Taxonomically, they are divided into three groups, namely Sarotherodon sp., Oreochromis sp. and Tilapia sp. They were distinguished from one another based on reproductive features. Sarotherodon sp. and Oreochromis sp. are mouth brooders whereas Tilapia sp. guards their nest. These cichlids fish are endemic to the freshwater habitat in Africa and domesticated for table purposes. The taxonomy of the species Oreochromis niloticus and Oreochromis mossambicus were shown in Table 1.1.
Table 1.1: Taxonomy of Oreochromis niloticus and Oreochromismossambicus Kingdom Animalia
Phylum Chordata Class Actinopterygii Order Perciformes Family Cichlidae
Genus Oreochromis
Species Oreochromis niloticus (Linnaeus, 1758) Oreochromis mossambicus (Peters, 1852)
Source: Integrated Taxonomic Information System (ITIS), (2015)
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morphology characteristics are similar which has a compressed body, cycloid scales and short upper jaw length, showing no sexual dimorphism. Dorsal spines and pelvic fin held by the bony spines. It also has soft ray parts at the continuous dorsal fin. The caudal fin provided with soft rays and it is truncated. During the spawning season, the pectoral, dorsal and caudal fins become reddish in colour.
The genital opening is located behind the anus which the morphology is different between genders.
Dorsal Spines
Mouth
Figure 1.1: General morphology of red hybrid tilapia (Oreochromis sp.) Pectoral Fin
Anal Fin
Pelvic Fin Dorsal Fin
The morphology of tilapia can be described as shown in the guideline in Figure 1.1 (Rakocy
,
2005; Srijaya et al., 2011)
. In tilapia species, the main1.2 Problem statements
Recently the production of tilapia in Malaysia was declined based on DOF report (DOF, 2015). This declination may be due several factors such as insufficient supply of seeds to local farmers and poor management of tilapia stocks in farm (Karuppanan et al., 2013; Mazuki, 2015). These situations lead to deterioration of physical appearance, growth performance and sensory quality which affecting the yield. These problems caused the existence of variability traits in tilapia stocks. Exploitation of the traits present in tilapia stocks through combination in breeding programme will discover tilapia with the economic traits for food fish.
1.3 Significance of study
Characterisation of tilapia morphometric is significant as a prerequisite for breeding programmes where selection requires variation between species and predicts the size of fish carcass. Evaluations of growth performance variation for these strains provide the information for profitable tilapia farming.
Complementary to the breeding goals, sensory attributes variation between strains may be exploited in the breeding programme. Generally, these works were carried out comprising of these elements aimed to produce red hybrid tilapia with good
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