Spermiogenesis is a conserved process across taxa but often adapts to species-specific fertilization strategies, influencing sperm morphology and traits like motility and longevity, which are crucial for reproductive success. Water frogs of the genus Pelophylax are notable for producing fertile hybrids, such as P. esculentus, which generate both haploid and diploid spermatozoa with diverse genome compositions. Despite this, little is known about the morphology and genome content of hybrid, and particularly diploid, sperm. This study used fluorescent in situ hybridization (FISH) with species-specific markers to analyze sperm from hybrid P. esculentus and its parental species, P. ridibundus and P. lessonae. Spermatozoa measurements revealed no significant differences in head and tail traits between haploid sperm from hybrids and parental species. However, diploid spermatozoa in hybrids were significantly larger than haploid spermatozoa, likely reflecting doubled DNA content. Correlations between head and tail lengths were observed in spermatozoa with P. lessonae genomes, suggesting potential functional implications. Our findings show that hybrids produce haploid sperm with morphology
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