Aedesaegypti is the most important vector for dengue, yellow fever and Zika viruses. Considering its medical importance, vector population control program utilizing radiation-based sterile insect technique (SIT) is one of the potential methods for preventing and limiting the dispersal of these viruses. The present study was undertaken to evaluate the dose-rates effects of γ-sterilization on quality parameters of sterile males. Males Ae.aegypti at the pupal stage were sterilized by applying 70 Gyγ-rays in varies dose-rates, i.e. 0 (control), 300, 600, 900, 1200 and 1500Gy/h utilizing panoramic irradiator. Adult males that emerged from the pupal stage were assessed for their quality parameters, which are the percentage of emergence, longevity, sterility and mating competitiveness. The results herein indicate that there was no major effect of dose-rate on the percentage of emergence, the data showedthat there were no differences between irradiated males compared with control. Generally, the longevity of irradiated males was lower compared to control. The data also demonstrated that longevity was significantly increased at the dose-rate from 300 to 900Gy/h, then decreased at the dose-rate 900 to 1500 Gy/h. Sterility of irradiated maleswas significantly different compared to control, while there was no significantly different at dose rate 300 to 1500 Gy/h. Mating competitiveness of irradiated males was increased at the dose rate from 300 to 1200 Gy/h, then the value was decreased significantly at the dose rate 1500 Gy/h. The dose-rate effects of γ-sterilization were discussed in the context genetic vector control, in particular, the SIT. The results give information and contribute to better understanding towards γ-sterilization optimization and quality parameters of sterile male Ae. aegypti on SIT methods.

Current Status and Recent Achievements of the Sterile Insect Technique Program Against Dengue Vector, Aedes aegypti, in Indonesia

Optimized conditions for the packaging and transportation of sterile males are crucial factors in successful SIT programs against mosquito vector-borne diseases. The factors influencing the quality of sterile males in packages during transportation need to be assessed to develop standard protocols. This study was aimed to investigate the impact of compaction, temperature, and duration factors during packaging and transportation on the quality of gamma-sterilized male Ae. aegypti. Aedes aegypti males were sterilized at a dose of 70 Gy, compacted into Falcon tubes with densities of 40, 80, and 120 males/2 mL; and then exposed to temperatures of 7, 14, 21, and 28 °C. Each temperature setup was held for a duration of 3, 6, 12, 24, and 48 h at a 60 rpm constant vibration to simulate transportation. The parameters of mortality, flight ability, induced sterility, and longevity were investigated. Results showed that increases in density, temperature, and duration significantly increased mortality and reduced flight ability and longevity, but none of the factors significantly affected induced sterility. With a mortality rate of less than 20%, an escaping rate of more than 70%, considerable longevity, and the most negligible effect on induced sterility (approximately 98%), a temperature of 7 °C and a compaction density of 80 males/2 mL were shown to be optimized conditions for short-term transportation (no more than 24 h) with the minimum adverse effects compared with other condition setups.

Background: For decades, insecticides have been central to controlling the yellow fever mosquito, Aedes aegypti (L.), but extensive use has driven resistance development. This study investigates resistance of Ae. aegypti to pyrethroids (permethrin, deltamethrin) and organophosphates (malathion, pirimiphos-methyl) and their underlying mechanisms across Malaysia, Thailand, Indonesia, and the USA. Methods: Adult female Ae. aegypti (3–5 days old, non-blood-fed) were subjected to World Health Organization (WHO) tube bioassays using 0.4% permethrin, 0.03% deltamethrin, 5% malathion, and 60 mg/m ² pirimiphos-methyl. Each assay included four replicates of 25 mosquitoes, with mortality assessed at 24 h post-exposure. Genomic DNA was extracted from 10 resistant individuals per population, and two coding regions of the voltage-gated sodium channel (VGSC) gene (domains II and III) were amplified and sequenced to detect known and novel kdr mutations. For biochemical analysis, 40 newly emerged, non-blood-fed females per strain were individually homogenized to quantify mixed-function oxidase (MFO), esterase (α- and β-EST), glutathione S-transferase (GST), and acetylcholinesterase (AChE) activity. Results: High resistance levels were recorded in Malaysian and US Ae. aegypti strains, with low mortality ranging between 9% and 22% for pyrethroids. New mutations T1520I (8–15%) and I1011M (10–15%) were identified in Malaysian populations, the first detection of T1520I in the country, while V1016I (10%) was newly detected in Indonesian strains. Malaysian mosquitoes had multiple kdr mutations (S989P, V1016G, F1534C, and T1520I) in triple- and quadruple-haplotype combinations. The US Riverside strain showed a nine- to 10-fold increase in β-EST and three- to fivefold increase in MFO and GST activity compared to the VCRU susceptible strain, indicating strong metabolic resistance. In contrast, the highly resistant Malaysian Hamna strain exhibited no significant upregulation ( P > 0.05) in detoxifying enzymes, suggesting resistance was driven primarily by kdr mutations. Thai strains lacked kdr mutations but exhibited altered AChE (20–35% remaining activity) and elevated GST (2–3 times higher than control). Conclusions: The detection of novel kdr mutations and diverse resistance mechanisms underscores the adaptability of Ae. aegypti to insecticide pressure and highlights the urgent need for continuous monitoring and integrated resistance management strategies. Graphical

Background and Aim: Rhodamine-B (Rh-B) marking shows a great potential for use in mark-release-recapture (MRR) studies for rear-and-release mosquito control strategies, including the radiation-based sterile insect technique. However, its applicability and evaluation in body-stain-irradiated males of Aedes aegypti have received little attention. The present study evaluated the use of Rh-B to mark gamma-irradiated male A. aegypti. Materials and Methods: Male A. aegypti were irradiated at the pupal stage at a dose of 70 Gy. After emergence, males were fed 0.1, 0.2, 0.3, or 0.4% Rh-B in 10% glucose solution for 4 days. Groups of unirradiated males that received the same feeding treatments were used as control groups. We evaluated the persistence of Rh-B and the longevity of males after Rh-B feeding. Furthermore, the use of Rh-B in irradiated A. aegypti for MRR experiments was evaluated at an urban site. Results: No difference was observed in the Rh-B persistence among all concentrations at the 24-h postmarking period ranging from 91.25 ± 1.61% to 96.25 ± 1.61% and from 90.00 ± 2.28% to 93.13 ± 2.77% for the unirradiated and irradiated groups, respectively. Rh-B persistence significantly decreased over time, and persistence was significantly longer with increased concentrations in both the unirradiated and irradiated groups. Longevity was considerably decreased by Rh-B feeding and irradiation. However, no significant difference in longevity was found among males fed various concentrations of Rh-B. Through MRR experiments, irradiated-Rh-B marked males were mostly detected within a radius of 20 m and 40 m from the center-release point. The mean distance traveled of the released males from the three MRR events was calculated to be 42.6 m. Conclusion: This study confirms that Rh-B body marking through sugar feeding is applicable for irradiated male A. aegypti, with only a slight effect on longevity. Furthermore, considering the significant reduction in persistence over time, further study is needed to assess the impact of this reduction on the calculation of field biological parameters resulting from MRR experiments. Keywords: irradiated males, mark-release-recapture, rhodamine-B, sterile insect technique.

Ernawan B, Surniyantoro HNE, Sugoro I, Tambunan USF. 2019. A mutant variant of Nix gene on γ-irradiated Aedes aegypti as a male-determination factor. Biodiversitas 20: 893-899. This study was aimed to examine the effect of γ-irradiation on the Nix gene sequence as a male-determination factor on Ae. aegypti. The novelty of the present study is the investigation on the gene mutation level of the deoxyribonucleic acid as an impact of ionizing radiation exposure. Males Ae. aegypti at the pupal stage were sterilized by applying 70 Gy γ-rays in varies dose-rates, i.e., 300, 600, 900, 1200 and 1500 Gy/h utilizing panoramic irradiator and males group without γ-irradiation as controls. Adult males that emerged from the pupal stage were assessed for the Nix gene sequence using a Real-Time Polymerase Chain Reaction method and were sequenced using Clustal Omega and Bioedit software, respectively. The results indicated that mutation occurred in Nix gene sequences were categorized as a point mutation. The change of Nix gene sequences was occurred between the nitrogenous base of guanine to adenine (G>A), thymine to adenine (T>A) and cytosine to adenine (C>A) in certain sequence numbers. There were six and nineteen on the forward-strand and reverse-strand of the Nix gene, respectively, due to γ-irradiation treatment. We conclude that γ-irradiation exposure causes point mutation on the Nix gene of males Ae. aegypti.