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Channa Marulius) in the Punjab, Pakistan SOME STUDIES ON REPRODUCTIVE BIOLOGY OF SOL (CHANNA MARULIUS) IN THE PUNJAB, PAKISTAN MUHAMMAD ZAFARULLAH BHATTI 03-arid-842 Department of Zoology Faculty of Sciences Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi Pakistan 2010 SOME STUDIES ON REPRODUCTIVE BIOLOGY OF SOL (CHANNA MARULIUS) IN THE PUNJAB, PAKISTAN By MUHAMMAD ZAFARULLAH BHATTI (03-arid-842) A thesis submitted in partial fulfillment of the requirement for the degree of Doctor of Philosophy In Zoology Department of Zoology Faculty of Sciences Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi Pakistan. 2010 CERTIFICATION I hereby undertake that this research is an original and no part of this thesis falls under plagiarism. If found otherwise, at any stage, I will be responsible for the consequences. Student’s Name: Muhammad Zafarullah Bhatti Signature:____________ Registration No.: 03-arid-842 Date: ____________ Certified that the contents and form of thesis entitled “Some studies on reproductive biology of Sol (Channa marulius) in the Punjab, Pakistan” submitted by Mr. Muhammad Zafarullah Bhatti have been found satisfactory for the requirement of the degree. Supervisor: ____________________ (Dr. Afsar Mian) Member: _____________________ (Dr. Mazhar Qayyum) Member: _____________________ (Dr. Azra Khanum) Chairman, Department of Zoology: ________________________ Dean, Faculty of Science: ________________________ Director, Advance Studies: _________________________ i DEDICATED TO MY PARENTS ii CONTENTS Page LIST OF TABLES VII LIST OF FIGURES X LIST OF PLATES XIII ACKNOWLEDGEMENTS XIV ABSTRACT XVI 1 INTRODUCTION 1 1.1. Background 1 1.2. Sol (Channa marulius) 8 1.3. Objective 13 2. REVIEW OF LITERATURE 16 2.1 Growth 16 2.1.1. Age Related Changes 2.1.2. Length Weight Relationship 17 2.1.3. Condition Factor 20 2.2. Karyotype 2.3. Reproductive Cycle 21 2.3.1. Gonadosomatic Index 22 2.3.2. Hepatosomatic Index 23 2.3.3. Histological Changes 24 2.2.3.1 Testes 25 iii 2.2.3.2. Ovaries 26 2.3.4. Steroid Hormone 28 2.3.4.1. Testosterone 29 2.3.4.2. Estrogen 30 2.3.4.3. Progesterone 31 3 MATERIALS AND METHODS 33 3.1. Study Area 33 3.2. Morphometric Analysis 40 3.2.1. Morphometric Measures and Meristic Counts 3.2.2. Length –Weight Relationship 3.3. Karyotype 3.3.1. Slide Preparation and Photography 3.3.2. Chromosome Analysis and Karyotype 3.3.3. Chromosome Classification 3.3. Growth 43 3.4.1. Seed Procurement and Transportation 43 3.4.2. Rearing 44 3.4.3. Sampling and Measurements 45 3.4.4. Analysis 46 3.5. Reproductive Changes 46 3.5.1. Sampling 46 iv 3.5.2. Histological Studies 48 3.5.3. Hormonal Profile 50 3.5.4. Analysis 51 4 RESULTS AND DISCUSSION 53 4.1. Morphometry 53 4.2. Karyotype 4.2.1. Chromosomes: Number and Morphology 4.3. Growth 73 4.3.1. Length 73 4.3.2. Weight 75 4.3.3. Length Weight Relationship 75 4.3.4. Condition Factor 78 4.4. Reproductive Biology 88 4.4.1. Gonadosomatic Index 88 4.4.2. Hepatosomatic index 96 4.4.3. Sex Hormone Profile 102 4.4.3.1. Estrogens 102 4.4.3.2. Progesterone 103 4.4.3.3. Testosterone 104 4.4.4. Histological Changes 111 4.4.5. General Consideration v SUMMARY LITERATURE CITED 116 vi LIST OF TABLES TABLE No. TITLE PAGE 1.1 World fisheries and aquaculture (excluding aquatic plants) 5 production and utilization (million tons) during different years. (The State of World Fisheries Aquaculture, FAO, 2008). 3.1 Monthly temperature range (o C) (maximum - minimum) 41 of north (Islamabad) central (Lahore) and southern Punjab, (Bahawalpur), between May 2005 and April 2007 (Source: Pakistan Meteorological Department) 4.1 Summary of different morphometric variables of adult 69 Channa marulius captured in December 2005 –February 2007 from Taunsa Lake (southern Punjab, Pakistan). lengths in “cm”, and weight in “g”, n = 21 for all variables 4.2 The summary of different meristic variables of adult 70 Channa marulius captured in December 2005 –February 2007 from Taunsa Lake (southern Punjab, Pakistan) n = 21 for all variables 4.3 Regression of different morphometric and meristic 72 characters against standard length in Channa marulius captured from Taunsa Lake (southern Punjab, Pakistan). All measure in “cm”. 4.4 Age related changes in length of Channa marulius 73 maintained in earthen ponds at Chenawan Fish Hatchery 4.5 Age related changes in weight of Channa marulius 77 maintained in earthen ponds at Chenawan Fish Hatchery 4.6 Age – length - weight relationships and condition factor 84 (K) in different age groups for Channa marulius at different ages maintained at Chenawan Fish Hatchery (n= sample size, K = Condition factor) 4.7 ANCOVA: output with age (month) taken as categorical 88 variable, length (log natural) as covariate and weight (log natural) as dependent variable in Channa marulius. vii 4.8 Values of gonadosomatic index (GSI) of female Channa 91 marulius during the sampling calendar months 4..9 ANCOVA output for the effects of months and length of 99 fish (covariate, ln) on ovary weight (ln) of female Channa marulius 4.10 Pairwise comparison among months based on estimated marginal means (means adjusted for the effect of covariate) on ovary weight of female Channa marulius 4.11 Values of gonadosomatic index (GSI) of male Channa marulius during the sampling calendar months 4.12 ANCOVA output for the effects of months and length (ln) of fish (covariate) on weight (ln) of testes of male Channa marulius 4.13 Pairwise comparison among months based on estimated marginal means (adjusted for the effect of covariate) on weight of gonads of male Channa marulius 4.14 Values of hepatosomatic index (HSI) of female Channa marulius during the sampling calendar months 4.15 ANCOVA output for the effects of months and length (ln) of fish (covariate) on weight (ln) of liver (ln) of female Channa marulius 4.16 Pairwise Comparison among months based on estimated marginal means (means adjusted for the effect of covariate) of weight of liver of Female Channa marulius 4.17 Values of gonadosomatic index (GSI) of male Channa marulius during the sampling calendar months 4.18 ANCOVA output for the effects of months and length (ln) of fish (covariate) on weight (ln) of liver of male Channa marulius 4.19 Pairwise comparison among months based on estimated marginal means (adjusted for the effect of covariate) on weight of liver of male Channa marulius 4.20 Values of estradiol of female Channa marulius during the sampling calendar months viii 4.21 Monthly differences in Estradiole level of female Channa marulius is analyzed by Analysis of Variance (ANOVA) i.e. significant at F (6, 26) = 63.596 , P < .05 followed by Scheffe test for multi comparison. 4.22 Values of progesterone of female Channa marulius during the sampling calendar months 4.23 Monthly difference in Progestrone level of female Channa marulius is analyzed by Analysis of Variance (ANOVA) i.e. significant at F (6, 26) = 183.56 , P < .05 followed by Scheffe test for multi comparison. 4.24 Values of testosterone of male Channa marulius during the sampling calendar months 4.25 Monthly differences in Testosterone level of male Channa marulius is analyzed by Analysis of Variance (ANOVA) i.e. significant at F (6, 26) = 49.212 , p < .05 followed by Scheffe test for multi comparison. 4.26 Frequency of ovaries under different maturation stages of female Channa marulius during different calendar months 4.27 Frequency of testes under different maturation stages of male Channa marulius during different calendar months 4.28 Matrix showing the Pearson correlation coefficient between different variables of reproductive activity in female Channa marulius (n = 33) 4.29 Matrix showing the Pearson correlation coefficient between different indicators of reproductive activity in male Channa marulius (n = 20) ix LIST OF FIGURES FIGURE NO. TITLE PAGE 1.1 World fisheries and aquaculture production and 6 utilization (The State of World Fisheries and Aquaculture - 2000 (SOFIA) 1.2 Distribution of Channa marulius in South East Asia 11 (Courtenay and Williams 2004; USGS) 3.1 The hydrographical map of Pakistan showing all rivers 40 and their tributaries and the densely populated Punjab province 3.2 Map of the part of Punjab province of Pakistan showing 45 main head works in the Punjab 3.3 Schematic representations of the body measurements 49 and head measurement of Channa marulius 4.1 PCA graph, Factor Score plots of total length for 78 different samples of the Channa marulius based upon 22 morphological characters. (F1- F21 are the fish specimen) 4.2 A karyotype of Channa marulius (Hamilton 1922) 81 4.3 Linear graph showing age related changes in length of 85 Channa marulius maintained at Chenawan Fish x Hatchery 4.4 Linear graph showing age related changes in weight of 89 Channa marulius maintained at Chenawan Fish Hatchery 4.5 Linear graph showing age Relationship between log 92 values of wet weight against the log values of total length of Channa marulius maintained at Chenawan Fish Hatchery 4.6 Linear graph showing age relationship between wet 93 weight against the total length of Channa marulius maintained at Chenawan Fish Hatchery 4.7 Graphical representation of condition factor for Channa 96 marulius over a period of 15 months maintained at fish Hatchery Chenawan 4.8 Bar chart of the b values (coefficient of variance) of all 97 groups of Channa marulius maintained at Chenawan Fish Hatchery 4.9 Pattern of seasonal variations in the gonadosomatic 100 index of female Channa marulius 4.10 Pattern of seasonal variations in the gonadosomatic 102 index of male Channa marulius 4.11 Pattern of seasonal variations in
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