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Thesis Issued 2026-09-08 EN

Identification of yy supermale using microsatellite dna markers in the production of genetically male tilapia

Author: Nargis Sultana

Abstract

In the culture of tilapia (Oreochromis niloticus), monosex male tilapia are preferred because it not only can prevent uncontrolled reproduction but also can increase production as male tilapia grow faster than females. Development of genetic markers and linkage maps have offered opportunities to understand the complex mode of sex determination which in turn can play significant role in monosex production. As a first step towards producing genetically male tilapia, neofemales were produced by applying the synthetic estrogen Diethylstilbestrol (DES) in three doses- 100 mg, 200 mg and 300 mg DES/kg feed. DES treatments resulted in 95% female while the rate of female in the nontreated group was only 55%. Among six DES treated females used for progeny test experiment, three families - HRT23, HRT26 and HTZF824 were found to follow a 3:1 (male: female) sex-ratio (p>0.05) which are indicative to families of 'XY' females. Fifteen microsatellite DNA markers putatively linked to the sex determining QTL were analyzed for molecular identification of male tilapia. Three markers UNH995, UNH985 and ARO172 showed some variation among different genotypes of male and female fish. Among the three markers ARO172 was found to be the most informative in differentiating male and female genotypes. Four offsprings of family HRT23 (‘XY' sex-reversed female) identified as YY male on the basis of marker analysis were verified by progeny testing. Upon crossing with normal females (XX), one of them produced 100% male offspring and the other three produced 88.40%, 96.84% and 86.36% male offspring, respectively. Thus we have produced and identified YY supermales that can give 100% or nearly 100% male progeny. We also have identified one molecular marker located in LG23 that can at least correctly differentiate YY males from normal male and female fish. This YY male can be used for large scale production of genetically all male tilapia. Further research however is needed to explain the variations in sex ratios in progeny of YY supermales. This would allow greater understanding of the apparently complex sex determining system of tilapia and stronger control over large scale production of genetically male tilapia for aquaculture.