How to cite
Uruku, N. M., & Ameh, O. R. (2026). Genetic Assessment of Fertilization, Hatchability and Survival Rate of African catfish (Clarias gariepinus; Burchell, 1822) Broodstock of River Donga Nigeria. International Journal of Research, 13(1), 158–170. https://doi.org/10.26643/eduindex/ijr/2026/10
* 1Uruku, Ndekimbe Mamndeyati and 2Ameh, Oyibinga Rose
1,2Department of Fisheries and Aquaculture, Federal University Wukari P.M.B 1020, Taraba State. Nigeria.
First Author – https://orcid.org/0009-0005-9645-8765
Second author – https://orcid.org/0009-0009-4244-8827
*Correspondence: uruksme@gmail.com
Abstract
The study on Breeding supernova of Clarias gariepinus genetic groups from River Donga Nigeria was carried out from February 2020 – March 2021 to investigate reproductive supernova among the genetic population of C. gariepinus of river Donga. Thirty (30) fish samples were utilised for the molecular analysis. DNA specimens were prepared for sequencing following standard laboratory procedure. Fish samples of the genetic groups were injected with synthetic hormone, after latency period they fish were strip of its eggs according to their haplotype’s groups, fertilized and were assessed for reproductive success and survival in the genotypes inbred (Hap1 ♀D x Hap1 ♂D and Hap3 ♀D x Hap3 ♂D) and crossbred Hap1 ♀D x Hap3♂D and Hap1 ♂D x Hap3 ♀D). The result reveal fertilization of 68.90±3.40 which was recorded in inbred haplotypes 3 (Hap3 ♀D x Hap3 ♂D) while higher hatchability of 54.03±7.23 was also observed in the inbreed of haplotype 3 (Hap3 ♀D x Hap3 ♂D) and survival of 91.71% in inbred of haplotype 1 (Hap1 ♀B x Hap1 ♂B) was recorded. Water quality parameters show positive correlation with reproductive indices. Therefore, the haplotypes, crossing method used in this research can be utilized to manage genetic resources and boost aquaculture production.
Keywords: Breeding; Crossbred; Genetics; Haplotype; Inbred; River Donga
Introduction
Breeding supernova, particularly in the context of aquatic species like Clarias gariepinus (African catfish), involves creating superior strains with enhanced traits such as growth rate, disease resistance, and environmental adaptability (Solomon et al., 2021). Genetic groups often consider both inbreeding, example; selecting for specific phenotypic traits such as faster growth or improved survival rate while risking inbreeding depression and outbreeding strategies, which can significantly affect the genetics and overall health of the populations, example; crossing local strains with genetically distinct populations to introduce new alleles that confer advantageous traits (Uruku et al., 2021). Proper management of broodstock is essential to maintain genetic diversity variability and avoid inbreeding depression, can adversely affect the performance of the offspring (Olaoye et al., 2020).
Fertilization in C. gariepinus typically involved spawning, were female broodstock release egg which are then fertilize by male milt. Successful fertilization is contingent upon several factors, including gametes quality, environmental condition such as water temperature and quality, and the timing of spawning event (Olaniyi & Omitogun, 2017). Various genetic strains may exhibit differences in these factors, leading discrepancies in fertilization rate (Ezenwo & Ajiboye, 2019). Following fertilization, the hatchability of eggs is influence by both genetic and environmental parameters. Factors such as egg viability, incubation conditions and care can all contribute to the success of hatching. Usman & Balogun, (2021) have indicated that some strains of C. gariepinus demonstrate higher hatchability rate due to enhanced genetic traits, leading to more viable embryos and improved survival rate.
Genetic groups within C. gariepinus can vary based on geographic location and breeding history. Genetic variation among strains of C. gariepinus can significantly influence reproductive outcomes. Diyaware et al. (2023) conducted a study using strains from River Benue and Gubi Dam, and the findings highlight the genetic potential of River Benue strains when involved in hybrid crosses.
Maintaining high genetic diversity is crucial for breeding programs to prevent the negative consequences of inbreeding. Using molecular tools like microsatellite or SNPs (Single Nucleotide Polymorphisms) can help in identifying genetic variation and structuring breeding programs effectively (Olaoye et al., 2020). Genetic markers are used to track the performance and health of breeding lines. The relationship between genetic strains of C. gariepinus with fertilization and hatchability is a complex interplay that holds significant implication for aquaculture. Advancing our understanding of these dynamics through this targeted research will enable the development of more robust fish stocks, there by supporting sustainable aquaculture and food security initiatives. Understanding how these genetic differences impact fertilization and hatchability is crucial for aquaculture producers aiming to optimize yield and improved sustainability (Ajayi & Ajani, 2020). Therefore, this study is aim to investigate breeding supernova (success) of C. gariepinus in river Donga, Taraba State Nigeria.
Materials and Methods
Description of the Sampling Site (River Donga)
Donga river lies between latitude 7°43′00″N and longitude 10°03′00″E. It has an area of 3,121 km² and a population of 134,111 at the 2006 census, figure 1. The Donga River is a river in Nigeria and Cameroon. The river arises from the Mambilla Plateau in Eastern Nigeria, forms part of the international border between Nigeria and Cameroon, and flows northwest to eventually merge with the river Benue, Nigeria. The Donga watershed is 20,000 square kilometres (7,700 sq mi) in area. At its peak, near the Benue the river delivers 1,800 cubic metres (64,000 cu ft) of water per second. A lot of fishing activities go on in the River and thus fishing is an occupation in the area (Inger et al., 2005).
Figure 1: Map showing locations of sampling site, River Donga a Tributary of river Benue
Procurement of fish samples
A total of thirty (30) broodstock each of C. gariepinus (average weight of 1000-1500g) both males and females were bought from artisanal fishermen. Fish were caught with various fishing gears at the River Benue, Taraba State, Nigeria. Gross physical examination of the external features of the samples were undertaken for abnormalities at the main landing site and samples obtained from the two Rivers were transported in plastic troughs (60cm diameter × 30cm deep) to Kahzuh integrated farm which is a leading modern Technological driven farm which lies on latitude 8º5′ 2.472ʺ N and longitude 9º47’34.008ʺ E in Gindin Waya, Ibi LGA, Taraba State Nigeria. It is bounded in the south by Benue state, North by Gassol LGA, East by Wukari LGA and West by Ibi LGA. Gindin Waya agro – ecological zone is the southern guinea savanna and it characterized by tropical hot/wet with distinct rainy and dry seasons. The hormone (Ovulin) was procured from Agro-service Centre Jos, Plateau State.
Broodstock Selection
Mature gravid females were selected based on swollen, well distended soft abdomen, reddish vent and gentle extraction of few eggs by pressing the fish abdomen using the finger. Females with sharp golden colored eggs were selected. Matured males were also selected based on their reddish pointed genital papillae. Only 30 matured specimens were utilized for this study.
Transportation/Acclimation of Clarias gariepinus Broodstock
The selected C. gariepinus broodstocks (male and female)were acclimatized in mobile holding ponds for a period of eight (8) weeks before artificially induced breeding were carried out.
Molecular Diagnosis and Separation of specimens to Genotypes
During acclimation period, caudal fin clip of about 1g each were obtained from each of the thirty (30) specimens of C. gariepinus and sent to IITA, Ibadan for gene extraction and genotyping by sequencing (GBS), to ascertain genetic divergence in the population. These were inferred by 16S rRNA primers. In order to achieve this, standard laboratory procedures of DNA extraction, polymerase chain reaction, cloning and sequencing were utilized it was then followed by bio-informatics analysis.
Experimental design
Both the parental and the intra – specific crosses were repeated three times in complete randomized block design (CRBD) manner, having hatchlings each after taking the pool weight and both were collected and stored in the aerated bowls. The survival of fry in each bowl per treatment were taken after egg yolk absorption.
Hormone preparation
Ovaprim: it does not require any special preparation. It was used to aid spawning in the reproductively matured female Catfish. Ovaprim (Western Chemical Inc. Femdale, WA) is marketed in liquid form and administered at the dosage of 0.5ml per Kg of each test brooder.
Administration of spawning agent (hormones)
The weight and length of the gravid female and male brooders were measured and induced with the hormones ovaprim at a dosage 0.5 ml/ kg/body weight for female following Efeet al.(2015).
Stripping of eggs from female brooders
The body of the female brooders were mopped dry and pressure was applied gently on the abdomen of the female brooders injected with spawning agent (Ovaprim). Ovulated eggs from the genital opening was collected in a plastic bowls with labels: strain-wise and weighed separately.
Fertilization and hatchability rate were estimated following Lambert, (2008) formulae.
Artificial fertilization of eggs
Spermatozoa (milt) from the mature male haplotype was used to fertilize the eggs in the labeled bowls in the following cross combination replicated in triplicate:
Experimental crosses
The following generic combinations were carried out:
Design for the Reproductive characterization
Haplotypes Location
River Donga
Haplotype 1 Equal number, equal size, equal sex ratio across the two locations
Haplotype 2 2
Haplotype 3 Equal number, size, sex ratio across the two locations as in Haplotype 1
2x2x2 Factorial Design for Breeding
Location Generic Groups
Inbred hap 1- Gen1 Inbred hap3-Gen 2 Crossbred hap1xHap3-Gen3 Recip Crossbred hap1xHap3-gen-4
Donga Hap1 FD X hap1MD Hap3 FD X hap3MD Hap 1FD X Hap3MD Hap1MD X Hap3FD
Hap = haplotype; Gen = Generic group; Recip = Reciprocal; MD = Male of Donga; FD = Female of Donga
Incubation of fertilized eggs
Incubation of the fertilized eggs were carried out in circular plastic bowls of 90cm diameter and 45cm depth with a carrying capacity of 120 litres of water each. The incubating tanks were interconnected flow through system and the fertilized eggs were spread in single layers on a net that was suspended in the incubating tanks to avoid overlapping of the eggs which could result in clogging. Hatching was observed between 18 – 28 hours. Both the parental and the reciprocal crosses were repeated three times in a complete randomized block design (CRBD) manner.
The water quality of the system of culture (hatchery unit) was monitored daily for: Temperature, pH, Dissolved oxygen, Ammonia (NH3) and Electric Conductivity. The analysis was done immediately after water samples collection. The parameters were determined insitu using a multi parameter water checker from the various hatching tanks;
Statistical analysis
Data on production and reproductive indices was analyzed using Minitab 14 software for descriptive statistics and Genstat Discovery edition 4 for analysis of variance (ANOVA) with respect to inbreed and their reciprocal crosses. Post hoc test was carried out using Duncan Multiple Range Test (DMRT) to determine the differences between the means (P=0.05) using SPSS version 20.0.
Results
Reproductive Success in the Haplotypes of C. gariepinus from river Donga
Results of the percentage hatchability, fecundity and percentage survival (Day 3) of fry of the inbreed and crossbred C. gariepinus from river Donga haplotype were as shown in Table 1. Equal weight of eggs was obtained for all the crosses. The highest percentage fertilization (68.90%) was recorded in inbred (Hap3 ♀D x Hap3 ♂D) and the lowest (45.86%) in inbred (Hap1 ♀D x Hap1 ♂D) strain. The highest percentage hatchability (54.03%) was recorded in inbred (Hap3 ♂D x Hap3 ♀D), followed by crossbred Hap1 ♀D x Hap3 ♂D (49.11%), and the least percentage hatchability of 39.51% was recorded in (Hap1 ♀D x Hap1 ♂D). The highest survival value of 91.71% was recorded in inbred Hap1 ♀D x Hap1 ♂D followed by crossbred Hap1 ♀D x Hap3 ♂D (90.53%), and the least percentage survival rate of 87.91% was recorded in Hap3 ♀D x Hap3 ♂D.
Table 1: Determination of Fertilization, Hatchability and Survival Rate of River Donga Haplotype
Hap1 ♀D x Hap1 ♂D Hap3 ♀D x Hap3 ♂D Hap1 ♀D x Hap3 ♂D Hap1 ♂D x Hap3 ♀D
Weight of eggs(g) 5.00±0.00 5.00±0.00 5.00±0.00 5.00±0.00
Estimated No of eggs 3000.00±0.00 3000.00±0.00 3000.00±0.00 3000.00±0.00
No. of fertilized eggs 1375.67±172.79 2067.00±110.94 1556.67±313.79 1940.00±184.29
No. of hatchlings 532.00±192.17 1112.00±94.00 768.670±195.68 922.33±164.20
%Fertilization 45.86±5.76 68.90±3.40 51.89±10.46 64.39±6.14
%Hatchability 39.51±15.82 54.03±7.23 49.11±3.40 48.38±13.35
Survival at Day 3 492.33±197.89 974.33±24.70 706.67±247.32 807.67±103.55
% Survival 91.71±3.58 87.91±5.23 90.53±8.35 88.07±4.84
Inbreed = (Hap1 ♀D x Hap1 ♂D – Haplotype 1 female crossed with Haplotype 1 male and Hap3 ♀D x Hap3 ♂D – Haplotype 3 female crossed with Haplotype 3 male).
Crossbreed = (Hap1 ♀D x Hap3 ♂D – Haplotype 1 female crossed with Haplotype 3 male and Hap1 ♂D x Hap3 ♀D – Haplotype 1 male crossed with Haplotype 3 female)
Table 2shows that among the water quality parameters of river Donga haplotype,temperature positively correlated strongly with only percentage survival at Day 3 at r = 0.79 and negatively correlated with number of fertilized eggs, number of hatchlings, percentage fertilization, percentage hatchability and survival at Day 3 at r = – 0.84, – 0.76, – 0.84, – 0.53 and – 0.73 respectively. pH shows positive correlation with number of fertilized eggs, number of hatchlings, percentage fertilization, percentage hatchability and survival at Day 3 at r = 0.89, 0.79, 0.89, 0.64 and 0.77 respectively and negatively correlated strongly with only percentage survival at r = – 0.94. Dissolved Oxygen correlated positively with number of fertilized eggs, number of hatchlings, percentage fertilization, percentage hatchability and survival at Day 3 at r = 0.51, 0.69, 0.51, 0.89 and 0.73 respectively and negatively correlated with only percentage survival at r = – 0.49. Ammonia strongly correlated positively with number of fertilized eggs, number of hatchlings, percentage fertilization, percentage hatchability and survival at Day 3 at r = 0.80, 0.90, 0.80, 0.98 and 0.92 respectively and negatively correlated strongly with only percentage survival at r = – 0.80. while electrical conductivity weakly correlated positively with all the water parameters.
Table 2: Correlations (r values) of Water Quality Parameters and Reproduction indices of Studied River Donga Population
Temperature (OC) pH D.0(mg/L) NH3(mg/L) Electrical Conductivity
No of fertilized egg -0.84 0.89 0.51 0.80 0.06
No of hatchlings -0.76 0.79 0.69 0.90 0.17
% Fertilization -0.84 0.89 0.51 0.80 0.06
% Hatchability -0.53 0.64 0.89 0.98 0.17
Survival at Day 3 -0.73 0.77 0.73 0.92 0.18
% Survival 0.79 –0.94 -0.49 -0.80 0.07
* Indicates that correlation is significant (P> 0.05).
Discussion
Breeding supernova of C. gariepinus
Breeding supernova in fish is a key focus in aquaculture as it directly affects productivity. The feasibility of crosses among the haplotype of C. gariepinus and its reciprocal cross-breeding was demonstrated in the present study. Fertilization and hatchability in this study is higher in value but similar in trend to the observations of Olaniyi & Akinbola, (2013) for C. gariepinus induced with Ovaprim (46.3%). The high hatching percentage observed in inbreed haplotype (Hap3 ♀D x Hap3 ♂D) 54.03% of river Donga C. gariepinus might be attributed to the genetic improvement through molecular diagnosis. The phenomenon of higher fertilization and hatchability in inbred versus crossbred C. gariepinus is complex and may be attributed to several factors, including genetics, reproductive biology and environmental adaptation (Yu et al., 2020).
Inbred population can sometimes exhibit greater genetic compatibility, especially if they have been selectively bred for desirable traits. This can lead to a higher rate of fertilization as gametes may be better suited to combine effectively (Fitzsimmons, 2000). Eknarth & Acosta, (1998) ascertained that inbred tend to have a more uniform genetic background, which can reduce the occurrence of incompatible gene interactions during fertilization and embryonic development. If the alleles controlling fertility traits are more consistent in inbred groups, this can result in higher fertilization rate.
Yu et al. (2020) revealed that where the genetic makeup of the female influences the success of fertilization and embryonic development can be more pronounced in inbred lines, leading to improved hatchability. If the inbred group have been adapted to specific farming conditions (such as water quality, temperature, or feeding regime), they may demonstrate better reproductive performance in those environments compared to crossbred haplotypes, which may be more variable in their response to environmental factors (Ajayi & Adesola, 2012).
It is however important to acknowledge that differences that arise from breeding history, age and water quality can affect hatching rates. Variations in seasons can also lead to differences in hatching rates, as rightly observed by de Graaf et al. (1995). So as long as fecundity does not drop, hatching rates and survival rates of larvae remain the key to viable and economically beneficial production of catfish fry and fingerlings.
The high survival rate of crossbreed haplotype of C. gariepinus during day 3 of rearing may be related to its hardiness and adaptation to environment. This is in agreement with Olufeagba and Okomoda (2015); Omeji et al., (2013) who reported high survival rate of local C. gariepinus reared under a medium stocking density for a short duration in protected tanks. Crossbreeding is used to achieve improved traits (heterosis), minimize inbreeding and obtain better hybrids (Jothilakshmanan & Karal Marx, 2013). Akankali et al. (2011) reported that apart from being able to obtain quality seed, the artificial propagation technique can also be used to develop strains superior to their ancestors by the methods of selective breeding, hybridization and molecular characterization.
Various factors related to water quality can influence reproductive success, growth and survival rates of aquatic species. Understanding the correlation between water quality and reproductive indices is essential for the management and conservation of aquatic ecosystems. Mean water parameters recorded shows positive correlation with the reproductive indices during the experimental period, temperature, pH, dissolved oxygen (DO) and ammonia were within the range of optimal levels for good growth and survival of C. gariepinus seeds.
Conclusion
Developing a breeding supernova for C. gariepinus combines aspect of genetics, aquaculture practices, and environmental consideration. Therefore, careful planning, selection, and management of genetic diversity can produce robust strains that will contribute to sustainable aquaculture production.
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