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. 2024 Jun 13;19(6):e0303352.
doi: 10.1371/journal.pone.0303352. eCollection 2024.

An oral toxicity assessment of a mosquito larvicidal transgenic algae (Chlamydomonas reinhardtii) using adult Zebrafish and its embryos

Fareeha Amjad  1 , Hamza Khan  1 , Muhammad Islam Khan  1 , Sidra Ayub  1 , Rashid Bhatti  1 , Rabbia Pervaiz  2 , Kausar Malik  1 , Mohsin Ahmad Khan  1
Affiliations

An oral toxicity assessment of a mosquito larvicidal transgenic algae (Chlamydomonas reinhardtii) using adult Zebrafish and its embryos

Fareeha Amjad et al. PLoS One. .

Abstract

Mosquito-borne diseases pose a global health threat, with pathogens like Malaria, Dengue fever, and others transmitted by mosquitoes. Our study focuses on evaluating the toxicity of genetically engineered mosquito larvicidal algae (Chlamydomonas reinhardtii) to non-target organisms, specifically Zebrafish. We conducted a 90-day experiment, feeding Zebrafish different combinations of larvicidal algae and commercial fish feed. Statistical analysis revealed no significant differences in mortality, allergenicity, or moribundity among groups. Hematology, molecular analysis, and necropsy showed no physiological differences. Our findings indicate that the transgenic algae (TN72.cry11Ba) had no adverse effects on adult Zebrafish or their larvae. This study confirmed the safety of algae on non-target organisms, such as zebrafish.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. PCR for the validation of cry11Ba gene.
Column 1: 100bp (Thermoscientific ladder), Column 2: 616bp PCR product of cry11Ba gene. Column 3: +ve control, Column 4: -ve control.
Fig 2
Fig 2. (A, B)Fertilized Zebrafish eggs before hatching (C, D) Zebrafish embryo at initial developmental stages (E, F) The morphology of Zebrafish organs and skin after dissection.
Fig 3
Fig 3. Liver tissues with H & E staining.
(A) represents the test group, (B) represents the control group. Scale bar = 100μm.
Fig 4
Fig 4. Ovarian tissues with H & E staining.
(A) represents the test group, (B) represents the control group. Scale bar = 100μm.
Fig 5
Fig 5. Intestinal tissues with H & E staining.
(A) represents the test group, (B) represents the control group. Scale bar = 100μm.
Fig 6
Fig 6. Zebrafish larvae at 32hrs, 40hrs, 56hrs, 64hrs, 72hrs and 96hrs respectively.
Fig 7
Fig 7. PCR to confirm there is no test gene integration in Zebrafish tissues.
Column 1: Shows 100bp Ladder (Thermoscientific). Column 2: Shows +ve control cry11Ba, Column 3: Shows -ve control wild type, Column 4: Shows Zebrafish intestine DNA, Column 5: Shows Zebrafish liver DNA, Column 6: Shows Zebrafish heart DNA.
Fig 8
Fig 8. Western blot for the confirmation of no gene integration in Zebrafish tissues.
Lane 1 shows positive control cry11Ba, Lane 2–4 are from experimental groups. Lane 5–7 are from control groups.
See this image and copyright information in PMC

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