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Genetic Variation Among Three Wild Populations of Stinging Catfish (heteropneustes Fossilis) by Allozyme Electrophoresis

Mohammad Arif Hossain
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Sharmin Suraiya
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Monjurul Haq
2Department of Fisheries and Marine Bioscience, Faculty of Biological Science and Technology, Jessore University ofScience and Technology, Jessore-7408, Bangladesh

Md. Mukhlesur Rahman Khan
Department of Fisheries Biology and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

Stinging catfish (Heteropneustes fossilis) is one of the most popular indigenous catfish having considerable potential for aquaculture and commercial importance in Bangladesh. With a view to assessing the genetic status of H. fossilis, three samples Chalan Beel (Pabna), Burungi Beel (Jamalpur) and Bagapura Beel (Mymensingh) were analyzed. For genetic variation study, five enzymes (LDH, EST, MDH, PGM and GPI) were used encoded by eight loci of which three were polymorphic (Mdh-1*, Est-1* and Gpi-1*). The highest mean proportion of polymorphic loci, mean number of allele and the mean proportion of heterozygous loci per individual of the Chalan Beel population were observed (25.00%, 1.250 and 6.250%, respectively). The highest gene flow (33.5) and lowest population differentiation (0.0074) found in Burungi Beel-Bagapura Beel indicated the close relationship among them. In the Nei’s UPGMA dendrogram, the Chalan Beel population formed one cluster by the genetic distance of 0.0371 and the other cluster was formed by Burungi Beel and Bagapura Beel populations (D=0.003). The results suggested that a considerable genetic variation is maintained among the natural H. fossilis populations.

Keywords: Allozyme electrophoresis, Stinging catfish, Heteropneustes fossilis, Genetic variation, Dendrogram

Location: Fisheries Biology and Genetics Laboratory, Bangladesh Agricultural University, Mymensingh

Start Date: 00-11-2007

End Date: 00-05-2008

Program Area: Animal Health and Management

Commodity: Cat fish

Objectives:

The objective of the present study is to assess the population structure of stinging catfish which may serve as an important part of the effort in the measures for the conservation and production of highly diversified stock of this species.

Methodology:

Research Laboratory: The experiment was accomplished in the Fisheries Biology and Genetics Laboratory, Bangladesh Agricultural University, Mymensingh. The equipment and other facilities available at the Fisheries Faculty were also used whenever required. The research study was conducted from November 2007 to May 2008. Sample collection: The experimental fish were collected from three different beels (wetlands) in Bangladesh. Allozyme study: Muscle samples were taken from each individual and stored at –18^oC until electrophoretic analysis. The gels were prepared using 40.8 g (12%) of hydrolyzed potato starch (STARCH-SIGMA-ALDRICH CHEME, Steinheim, Germany) in 340 ml of distilled water and appropriate buffer. In this way, at first the starch powder was weighed by electronic balance (METTLER TOLEDO, PG503-SDR, Switzerland) and was transferred into a 1L Erlenmeyer flask containing 17ml (1/20 of total liquid volume) of electrophoresis gel buffer (CA 6.1). Then 323 ml of distilled water (19/20 of total liquid volume) was added to the mixture and immediately swirled to generate a uniform suspension and finally was heated to boil for 8-10 min until transparent solution of the gel was observed using a Bunsen burner. Then the boiled gel solution was degassed for approximately one minute to remove air foam by gentle shaking of the flask. The boiled starch was then poured onto the glass (18 x 22 x 0.5 cm³) attached with plastic frame (16 x 20 x 0.5 cm³). The hot gel was allowed to cool at room temperature for about 2 hours. The gel was covered with OHP sheet to prevent desiccation of gel after 30 min. Finally, after cooling the gel was preserved overnight into a refrigerator at 5-8⁰C to increase hardness for easily slicing of the gel. Enzyme were assessed using horizontal starch gel electrophoresis in one buffer systems (CA 6.1) and gel staining procedures were based on the methods of (Shaw and Prasad, 1970). The enzymes examined were as follows: Esterase (EST; EC 3.1.1.1), Glucose-6- phosphate isomerase (GPI; EC 5.3.1.9), Lactate dehydrogenase (LDH; EC 1.1.1.27), Malate dehydrogenase (MDH; EC 1.1.1.37) and Phosphoglucomutase (PGM; EC 5.4.2.2). After staining, the gel slice were preserved using cellophane paper, 10% acetic acid and 10% glycerin. At first, the staining gel washed with 10% acetic acid kept in 10% glycerin for 10- 15 min. Finally, the gel was kept on the cellophane paper and covered by other cellophane paper. Then the gel was kept in oven at 50 to 55^oC since over night for drying the gel. Genetic data analysis: Allelic frequencies were inferred directly from observed genotypes. Hardy-Weinberg equilibrium of genotype frequencies was tested using a chi-square test. When the most common allele existed in a frequency less than 0.95 at a given locus, this locus was regarded as polymorphic.

Source of Information: Journal of Fisheries, Volume 1 Issue 1 Pages: 14-21 December 2013

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The result of the present study would be useful to know genetic variation and structure of different populations of the studied species before undertaking any stock improvement and conservation program. To discuss the detailed population structures over time, further studies are required dealing with a large number of populations intensively sampled from different parts of the country. Moreover, large sample size and more enzymes are required. This information would help to identify distinct population groups existing in Bangladesh and consequently enables to formulate genetic conservation plan. Moreover, it may show the impacts of brood stocks management practices. However, there are few draw backs in the present study. Due to limitation of highly sophisticated equipment and reagents, it was not possible to analyze huge number of individuals per stocks. We screened 30 individuals from each of the stocks and numbers of enzyme were only five, which is not sufficient to draw a conclusion about the genetic structure of the three stocks of stinging catfish in Bangladesh. Further studies involving large number of samples and more loci are needed to have a precise knowledge about the genetic structure of this valuable fish species.

Knowledge Management: Journal