Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 Jun 23;51(4):114.
doi: 10.1007/s10695-025-01529-4.

Ameliorative potential of dietary supplements, ZnO-K, citrus essential oil, and pumpkin seed oil, on sperm quality in Nile tilapia: Insights from CASA, DNA integrity, antioxidant enzymes, and gene expressions

Marwa M Ali  1 , Kamal Fathy Elboray  2 , Engy T Megahed  3 , Hany T Abu-Taleb  3 , Alshimaa E Elsayed  1   3 , Eman Y Mohammady  4 , Mona S Amer  1   3 , Soliman A Morsi  1   3 , Eman M Abbas  3 , Mohamed S Hassaan  1 , Hosam Elsaied  5
Affiliations

Ameliorative potential of dietary supplements, ZnO-K, citrus essential oil, and pumpkin seed oil, on sperm quality in Nile tilapia: Insights from CASA, DNA integrity, antioxidant enzymes, and gene expressions

Marwa M Ali et al. Fish Physiol Biochem. .

Abstract

Improving sperm quality is essential for enhancing the reproductive efficiency of Oreochromis niloticus. This study evaluated the effect of three dietary antioxidant supplements, kaolinite-doped zinc oxide (ZnO-K), citrus essential oil (CEO), and pumpkin seed oil (PSO), on sperm quality parameters. A suite of integrative assessments, including Computer-Assisted Semen Analysis (CASA) parameters, spermatozoa DNA integrity, antioxidant enzyme assays, and gene expression profiling, was applied to validate sperm quality. A total of 192 adult males (mean weight 421.31 g ± 30.41 SD) were assigned to four dietary groups, each with three replicates. The control group received a basal diet without supplementation. The experimental groups were fed diets supplemented with ZnO-K (0.06 g kg⁻1), CEO (10 g kg⁻1), or PSO (15 g kg⁻1). ZnO-K supplementation significantly (P < 0.05) increased milt volume (1.40 ± 0.10 ml) and sperm concentration (5.676 × 109 sperm ml-1), along with marked improvements in CASA parameters (sperm motility and velocity) and DNA integrity. Antioxidant enzymes, catalase, CAT, glutathione peroxidase, GPX, and superoxide dismutase, SOD, were also significantly elevated in the ZnO-K group, recording activity values of 47.333 ± 1.452 U ml-1 milt, 65.667 ± 5.547 mU ml-1 milt and 60.667 ± 3.382 U ml-1 milt, respectively. Gene expression analysis revealed that CEO induced the highest upregulation of HSP70, while both CEO and ZnO-K significantly upregulated CC chemokine expression compared to control. In contrast, PSO consistently showed the lowest performance across all measured parameters. Collectively, ZnO-K supplementation most effectively enhanced sperm physiological and biochemical quality, whereas CEO predominantly supported cellular stress protection through gene expression modulation. These findings offered practical implications for optimizing artificial reproduction and sperm cryopreservation strategies in tilapia aquaculture.

Keywords: O. niloticus; Antioxidant supplements; CASA parameters; DNA integrity; Enzyme bioassay; Gene expression; Sperm quality.

PubMed Disclaimer

Conflict of interest statement

Declarations. Competing interests: The authors declare no competing interests. Authentication: The experimental fish handling protocol was approved by the National Institute of Oceanography and Fisheries (NIOF) committee for institutional care of aquatic organisms and experimental animals, under the no. NIOFAQ5F25R009.

Figures

Fig. 1
Fig. 1
CASA for sperm DNA integrity. A and B, enlargements of sperm heads. N, the head of sperm shows non-fragmented DNA, with a halo. D, head of sperm shows fragmented DNA, without halo. Scale bar = 5 µm; bar in A or B = 1.5 µm
Fig. 2
Fig. 2
Sperm morphological structures for males fed on diets supplemented with different antioxidants, 1. Head sperm, 2. Midpiece and 3. Tail. Scale bar = 10 µm
Fig. 3
Fig. 3
a Effect of the supplements ZnO-K, citrus essential oil, and pumpkin seed oil on the gene HSP70 expression in the sperms of O. niloticus. Bars with different superscript letters (a, b, c, d) illustrate significant differences (p < 0.05). Error bars +/- 1 SD. b Effect of the supplements ZnO-K, citrus essential oil, and pumpkin seed oil on the gene CC chemokine expression in the sperms of O. niloticus. Bars with different superscript letters (a, b, c) illustrate significant differences (p < 0.05). Error bars +/- 1 SD
See this image and copyright information in PMC

References

    1. Abascal FJ, Cosson J, Fauvel C (2007) Characterization of sperm motility in sea bass: The effect of heavy metals and physicochemical variables on sperm motility. J Fish Biol 70:509–522
    1. Agarwal A, Durairajanayagam D, Halabi J et al (2014) Proteomics, oxidative stress and male infertility. Reprod Biomed Online 29:32–58 - PubMed
    1. Aghaie S, Nikzad H, Mahabadi JA et al (2016) Protective effect of combined pumpkin seed and ginger extracts on sperm characteristics, biochemical parameters and epididymal histology in adult male rats treated with cyclophosphamide. Anat Sci Int 91:382–390 - PubMed
    1. Ahamed HA, Mohamed MJ, Arunachalam KD et al (2020) Effects of Azomite enriched diet on gonadal steroid hormone levels and milt quality indices in Oreochromis mossambicus. Aquac Rep 17:100341
    1. Alavi SMH, Psenicka M, Rodina M et al (2008) Changes of sperm morphology, volume, density and motility and seminal plasma composition in Barbus barbus (Teleostei: Cyprinidae) during the reproductive season. Aquat Living Resour 21:75–80

LinkOut - more resources