EFFECT OF ALBINO GENE ON INTRASPECIFIC HYBRIDIZATION OF NORMAL AND ALBINO AFRICAN CATFISH (CLARIAS GARIEPINUS)

ABSTRACT

The effect of albino gene on intraspecific hybridazation of African Catfish and Albino Clarias gariepinus was carriedout to determined their latency period, fecundity, fertility, hatchability survival,  growth  performance,  some  morphometric and  meristics  characteristics  and  genetic characterization of parents and offspring of F1 generation. A total of four (4) mating combination experiments were conducted using eight (8) ripped, matured normal and albino Clarias gariepinus with average body weight of 1.64kg. The broodstock were purchased from Abdulfana fish farm Yola and transported in 50 litres jerican to the Department of Fisheries and Aquaculture Adamawa State University Mubi. They were acclimatised for 10 days, vital feed were fed daily at 5% of their biomass. Artificial breeding was cariedout using Ovaprim hormone at a dose of 0.50ml/kg and 0.25ml/kg for female and male respectively. After hand stripping of the female brooders, nine hundred and sixty (960) eggs were used for fertilization in each mating combination, 320 eggs replicated three times using Randomized Complete Design (RCD) to determine the percentage Fertility and Hatchability of the eggs. Two hundred and forty fry of two weeks old from each treatment were randomly selected and stocked in same culture system and fed with Coppens feed at 10% body weight, the experiment lasted for the period of 18 months for both F1 ans F2 gerations, where weight, length and survival/mortality rate, in each treatment and replications were recorded weekly.Water quality parameters were monitored weekly. Gene characterization (DNA extraction, Electrophoresis, Selection of Microsatellite Markers, Polymerase Chain Reaction (PCR) Amplifications and Electrophoresis for PCR, was carried out on the parent stock and all the offspring of mating combination in F1 generation. It was observed that female albino Clarias gariepinus had the highest latency period of 13.50±0.22 hours at 27.50±0.20oC and less fecundity of 4032±0.12, while the normal Clarias gariepinus had the least latency period of 12.00±0.21 hours and high fecundity of 7020±0.23 at similar temperature. Similarly crossed between NN♂ x NN♀ had the highest fertility and hatchability of 44.79±0.32% and 72.6±0.24%, while the least value of 27.92±0.31% and 46.3±0.43% were recorded in AA♂ x NN♀. Also crossed between NN♂ x AA♀ had the highest final mean weight gain of 912.90 ±0.00g, followed by NN♂ x NN♀ with 900.75 ±0.00g, while NN♀ x AA♂ and AA♂ x AA♀ had the least value of 893.88±0.00g and 852.98±0.00g respectively. High survival rate of 41.25±0.00% was recorded in crossed between AA♂ x AA♀, while NN♂ x AA♀ had the least value of 12.50±0.00%. The Head width, eye diameter and occipital fontanelle of the F1 generation was higher than that of the parents. All the water quality parameters measured during the course of the research were within the normal range for tropical catfish cultured. The two loci of normal Clarias gariepinus (Cg01) and albino Clarias gariepinus (Cg02) showed amplification. The size of the two (2) microsatellite loci ranged from 0.92 to 1680 bp, with low estimates of null allele frequencies across all populations for the parent stock and 0.93 to 1.00 for F1 generation. The number of Allele observed from the genetic analysis of parent stock and F1 generation of Cg01 showed heterozygosity and the second allele on the second band Cg02 shows homozygosity. However, a homozygote excess was estimated in the parents of Cg02 and the result was 100% in both the parent and F1 generation of the normal and albino Clarias gariepinus. Normal male and female Albino Clarias gariepinus can be use by fish farmers to produced hybrid that have high survival rate and can grow faster. Further  study should be carriedout to determined the causes of had skin, large head width,  large eye diameter and  wide occipital fontanelle of the F1 generation.

CHAPTER ONE

1.0  INTRODUCTION

1.1 BACKGROUND OF THE STUDY

The demand for high quality protein especially from aquatic resources is rising, Clarias gariepinus is the most common and have received much attention because of its economic importance and high rate of success in rearing them (Adewolu et al. 2009). Albinism in catfish is the best known potentially valuable qualitative traits and inherited as a single homozygous recessive trait (Bondari, 1981). The term albino catfish can refer both to an albino specimen of any catfish species or to a particular species of catfish the Albino Clarias gariepinus Catfish (Westerman and Birge, 1978). Albino specimens of all catfish species including the Albino Clarias dramatically, increased agricultural production is clearly needed to meet this demand in the future, because capture fisheries is showing precipitous disclose due to over fishing habitat, destruction and increasing population and increased in capture fisheries are not anti-coopted under the current global condition (Dunham et al. 2001) the most current advance in aquatic production have been achieved through the application of genetic principles which includes selective breeding, hybridization, chromosome manipulation, sex reversal and gene transfer (Aliah and Taniguchi 1999). Framed sources are economically important food fish which have gained much prominence species of characterized by hardiness, resistance to diseases, high yield potentials and ability to grow on a wide of nature and low cost ratification food and can withstand low oxygen and wide pH levels (Fagbenro and Sydenham, 1991). Genetic techniques have been applied to other animal to improve their qualify but that of fish is not yet fully exploited. Failure to pay attention to the genetic aspect of fish husbandry may result to decline in productive as inbreeding or other genetic consequences of mis-management may result in slower growth, decreased viability and disease susceptibility as well as decreased egg production (Bartly et al. 1990). Hybridization has been found to be a breeding programme that tries to find mating combination between different population of fish which produce hybrid vigor (tave, 1993). Hybridization has been used to increase growth rate, manipulate sex ratio, produce sterile animals, improve flesh quality, increase resistance and to tolerant to environmental extremes as well improve a variety of other     traits that make other aquatic animals production more profitable (Dunham  et al. 2001).  Intraspecific hybrids are produced when useful traits in parent fishes are transferred to the offspring, this is because for back cross hybrid produce from the fertile hybrid could not be easily differentiated from the pure parents (Aluko, 1996). The hybrid cross between Heterbranchus and Clarias Species is receiving attention in Africa particularly Nigeria this hybrid had been repeated to show that Heterobranchus species usually grow larger than the Clarias species,therefore shows better percentage survival (Madu Clarias spcies in order to assess the aquaculture potentials of their hybrid s in terms of growth performance and survival compared to the putative parents at the fry, fingerlings and adult stage (Onyia,  2010).  Clarias  gariepinus  is  an  economically important  fresh  water  species  of  the Clariidae family that contribute immensely to the annual fresh water fish production in Nigeria. They are also readily acceptable among Nigerians fish farmers and consumers, hence command high commercial value (Adebayo et al. 2001). They are commonly referred to as mud fishes or African catfish in various parts of Nigeria and are important source of animal’s protein. Among the freshwater species for culture in Nigeria gariepinus is a natural occurring phenomena and that there has always been albino Clarias gariepinus even if only a few survived to become adults. An albino specimen of any of the larger catfish species can be a truly magnificent sight, such as large albino catfish (Groblan et al. 1992). The inheritance of albinism has been analyzed in many aquarium fish and several aquaculture species-Rainbow trout, channel catfish and Grass carp (Delgado et al., 2009). In all the traits of the regular clarias gariepinus plate i and ii. They have the same well testing meat, and they grow up to the same size (Britton and Davie, 2006). However, the albino form of this catfish has some additional values. The Albino from of the catfish is an appreciated aquarim fish and can be sold in large quantities to aquarium stores (Galib et al., 2012). Albino catfish are also preferred by some to stock fishing lakes with, since the albino catfish are unsuccessful in catching any (Clark, 2002). There is no doubt that commercial breeding of catfish in captivity has increased the amount of Albino catfish available both in the aquarium trade and in the wild as a result of different restocking efforts (Piorski, 2010). Albinism is passed genetically from parents to offspring (Dobosz et al. 1999). Each cell contains numerous pairs of genes, one from each parent. These genes transmit traits through generations. An albino offspring results from a specific combination of genes (Rothbard and Wohlfarth 1993). Albinos are not in frequent because the genes for that trait are recessive, while the genes for normal pigmentation are dominant (Rothbard and Wohlfarth, 1993). If both are present, normal pigmentation occurs. If only recessive genes occur, albinism may result. Only a small percentage of animals carry the recessive gene, so the chance of the pairing of recessive genes in an individual animal is slight. In natural environment, there are several reports of total or partial albinism in freshwater fishes and marine fishes (Piorski, 2010). Some cases of total or partial albinism in siluriformes have been reported, e.g., Ictalurus punctatus (Westerman and Birge, 1978), Schizolecis guntheri (Brito and Camaraschi, 2005). Intraspecific hybrids are produced when useful traits in parent fishes of the same genus are mated. Intergeneric hybrids on the other hand are generated when parents from two genera are crossed. All of the genetic information in an individual fish is contained in molecules  of  deoxyribonucleic  acid.  Molecules  of  DNA  are  composed  of  sub  units  called nucleotides  in  DNA because there  are  four different  bases  (adenine,  guanine, thymine  and cytosine). DNA molecules consist of a long ladder of paired nucleotides. The helix in the ladder gives the DNA molecule a double helix structure. Nucleotides form base pair in the double helix in a specific manner where thymine is found in one strand of the helix, only adenine will be found in the same position of the opposite strand. Similarly, where guanine is found in one of the chrosome strands, only cytosine will be found in the same position of the opposite strand. The two strands of the helix are said to be complementary because of the way nucleotides form base pairs (Westerman and Birge, 1978). Whenever a cell divides, the DNA must be replicated in order to provide each daughter cell with a complete set of genes. An advantage of complementary base pairing is evident during replication of the DNA molecule. During replication, the two strands of the DNA helix are separated by enzymes so that each strand is available to serve as a template for a new molecule. Individual nucleotides are affixed to each template. Two complete and identical DNA molecules resulted, the complementary pairing of bases ensures that the replication of DNA is essentially error free. Microsatellites represent another type of repetitive DNA (Westerman and Birge, 1978).

2-6-base units (motifs)., the allelic variability of microsatellites is based on the difference in the length of fragments caused by the different number of repeated units. The isolation of microsatellite loci from a genomic library for a new species or group of species is the first step for the investigation of microsatellite polymorphism (Calcagnotto et al. 2001). The sequence information for the flanking DNA is needed to allow the synthesis of specific PCR primers. Following primer design, products can be amplified and separated through capillary electrophosatellites are locus-specific and co dominant markers (Calcagnotto et al. 2001).

30. Heterozygote for microsatellite loci has two bands on a gel which represent two alleles while homozygote has one band (one allele). Microsatellite alleles are designated by numbers which indicate the size of corresponding PCR product in base pairs (Taniguchi et al. 1999) the first studies on application of microsatellites as genetic markers in fish were published in the beginning of the 1990s. From this time, microsatellites were identified and analyzed in many important fisheries and aquaculture species (Sirol and Britto, 2006). Funkenstein et al. (1990) documented that very high variability, discrete character of inheritance, co dominance, and established standard techniques for their analysis, microsatellites are regarded now as the most popular and powerful type of DNA markers in fish genetics.

1.2    Statement for the Research Problem

In Nigeria and other developing country, there is little information on the effect of hybridization of Albino and Normal catfish Clarias gariepiunus the hybrids have been reported to show heteroses (Salami, et al. 1993, Aluko, 1999) but their suitability for aquaculture has not been thoroughly evaluated. There is also little information in literature on the growth and viability, hatchability, latency period, fecundity, fertility, fertilization, morphometric and meristic characters and colour ratio of the mating combination as well their gene characterization of the intraspecifichybridization of normal and albino Clarias gariepinus, The work of Onyia et al. normal Clarias gariepinus grow faster than the Albino C. gariepinus, hence there is need to assess the viability, latency period, fertility, fecundity, morphometric and meristic character, color and sex ratio as well their genetic characterization of the intraspecific hybrids of normal and Albino Clarias gariepinus in F1 and F2 which were lacking in their reports.

1.3       Justification for the Study

Production and culture of species of the catfish belonging to the Claridae family is fast growing globally according to (Adewolu et al. 2008). This study will be useful in assessing viability, growth performance, latency period, fertility, fecundity, fertilization, colour and sex ratio of the various intraspecific mating combination of Albino and normal Clarias both in F1and F2.

This study was initiated to provide detailed phenotypic appearance (morphometric and meristic character) description of the intraspecific combination of Albino and normal Clarias gariepinus in order to know the distinguish features for identification of their hybrids and their putative parent. This study on intraspecific hybridization of Clarias gariepinus Albino and normal and their reciprocal  hybrids  is  of  great  importance in  ensuring  a proper understanding of their  gene characterization in both parent and F1 (Betiku and Omitogun, 2006) to create a good foundation for producing a genetically improved hybrid fish.

1.4   The Aims of the Study

The Aim of this study was to determine the effect of albino gene on intraspecific hybridization of normal and albino african catfish (Clarias gariepinus).

1.5    The Objectives of the Study

i.        to determine the latency period of the female parent stocks of normal pigmented, albino Clarias  gariepinus and the female of hybrids in (F1) generation of their crosses.

ii.      to evaluate the fecundity, fertility, and hatchability of eggs of different mating combination in (F1 and F2) generation of Clarias gariepinus.

iii.       to determine the percentage survival of F1 and F2 generation of Clarias gariepinus.

iv.        to determine the growth performance and feed utilization of F1 and F2 generation of the hybrids.

v.         to determine the morphometric and meristic characters of the F1 and F2 generation of Clarias gariepinus.

vi.       to access the genetic characterization of the parent stocks and F1 generation

1.6        Research Hypotheses

i.         There is no significant differences (P>0.05) between the latency period and fecundity, of the parent stock and F1 of normal and albino C. gareipinus.

ii.         There is no significant differences (P>0.05) between the percentage fertility, fertilization, hatchability and survival of normal and albino Clarias gareipinus in F1 and F2.

iii.        There was no significant differences among the percentage survival of the offspring in F1 and F2.

iv.       The growth performance and feed utilization of normal and albino Clarias gareipinus in

F1 and F2 doesnot differ.

iv.        The morphometric, meristic and genetic characterization of the parent stock and their hybrid of normal and albino Clarias gariepinus doesnot differ.

vi.        The phenotypic and genotypic appearance of hybrid of normal and albino Clarias gariepinus do not differ.

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