Green Silver Nanoparticles Effects on The Susceptible and Field Strains of The Housefly, Musca domestica L. (Diptera: Muscidae)

Musca domestica , commonly known as the house fly, is critical because it is a major vector for several infectious pathogens, including those that cause cholera, typhoid fever, dysentery and some skin diseases, in addition to trachoma and trachoma considered a household pest and a public health hazard (Greenberg, 2019). House flies thrive in unsanitary environments such as slaughterhouses, dairies, and ranches that encourage the spread of disease. Pathogen transmission can occur when insects come into contact with human habitats (Khan et al., 2012).

Although the systemic application of chemicals has brought progress, the dangers posed by chemicals, such as the loss of beneficial arthropods, environmental imbalances, contamination of manure, the environment, freshwater, food, and employees, are quickly becoming apparent. These factors have supported pest resurgence and contributed to the rapid emergence of resistant individuals, leading to irreversible environmental and health hazards such as cancer, genetic aberrations, poisoning and death (Prabhaker et al., 1998). House flies are known for their ability to evolve resistance strategies to evade and detoxify chemical pesticides. Resistance to organophosphates, carbamates, and pyrethroid insecticides has been documented (Boxster and Campbell, 1983; Kaufman et al., 2001; Butler et al., 2007).

                Recently, pesticide formulations have been developed using nanotechnology techniques (Abd-Elnabi et al. 2021). As nanoparticles exhibit new properties, nanotechnology has emerged as one of the most promising new approaches to pest control in modern agriculture and is expected to revolutionize the field of pest control in the near future. (Bhattacharya et al., 2010, Rai et al., 2018, Tunçsoy, 2018, Ahmed et al., 2019, Khoshraftar et al., 2019, Shahzad and Manzoor, 2019)

Metallic nanoparticles produced by various chemical processes are of interest applications of biomedical worldwide. These methods reveal a wide range of toxicities in non-target organisms. The biosynthesis of NPs is an environmental safety manner without the use of toxic chemicals, and increasing the safety of environment in the future. Therefore, the development of green synthesis of Ag NPs has emerged as an important area of ​​nanotechnology, and the use of biological entities such as plant extracts and biomass, as well as microorganisms for the production of NPs, can be used both chemically and physically. In addition, conventional methods were   safety to the environment (Reddy et al. 2012). Advances in green synthesis beyond physical and chemical methods are environmentally friendly, inexpensive, and easily scaled up for large scale synthesis of NPs, but green synthesis involves high temperature, energy, pressure and hazardous no chemicals are required (Ahmed et al. 2016).

El Monery et al. (2021) found that its components such as lignin, pectin and hemicellulose can be used for the reduction of silver ions to produce silver nanoparticles Ag NPs. Therefore, the main aim of this study is evaluate the efficacy of green synthesized from silver nanoparticles as alternative insecticides on the two experimental houseflies strains (Diptera: Muscidae) susceptible and resistant in the laboratory conditions.

MATERIALS AND METHODS

Insect rearing

The house fly, Musca domestica used during this study was reared in the laboratory of Economic Entomology in Plant Protection Department, Faculty of Agricultural (Saba Basha), Alexandria University  at 28 1 C, 60% 10 R.H. and 18 hrs illumination. Adults of a susceptible and field strains of M. domestica were maintain cages in 40  cm, with a plywood floor and wooden frame. Larval, adult, oviposition media were prepared according to Singh and Jerram (1976).   

The preparation of starch paste, silver nitrate solution and silver nanoparticles

Prepared amount of Na OH 0.3 g then dissolve it in water, and lye is added gradually to a known amount of crude rice starch 2 g while stirring. This prepared mixture was stirred continuously until the starch was completely dissolved. The pH of the prepared solution reached to 11 and its temperature reached to 60 °C. After that, the previous solution was used for preparing different concentrates of silver nitrate 0.25 and 0.5 g then all concentration was dissolved in 100 ml of distilled water. After that, the different concentrations of the silver nitrate solution which prepared was dropped to the starch solution which stirring at 60 ^oC and pH 11. After 15 minutes, the silver color was changed gradually from dark white to transparent yellow. Then the silver nanoparticles Ag NPs were formed and the color intensity of Ag NP changed due to the amount of silver nitrate added. So, the darker color means the higher concentration of Ag NPs formed. Based on (El-Rafiea et al 2013 and Abdelsalam et al 2019) they have two concentrations of Ag NP denoted Ag NPs-2000 and Ag NPs-4000 ppm

The characterization description of silver nanoparticles Ag NPs

In this experimental work the characterization of silver nanoparticles were used according to (Abdelsalam et al. 2019) description.

Bioassay of housefly:

In the laboratory 1 ml of AgNPs was taken from different concentrate (10, 20, 30, 40 and 50  ) and mixed with 20 ml of larval media into plastic cup, each concentration included three replicates each of them had 10 larvae in third stage of age. In addition, the compared larvae in control (C) were feed with 20 ml of media mixed only with 1 ml of distilled water. The experiments were carried out in third generation at field strain and eighth in sensitive strain. The ratio of mortality calculated by counting the number of dead larvae after 24, 48, 72 hrs and trace the pupal weight, inhibition of pupation, adult emergence, sex ratio and other biological aspects were appeared. (Abd El-Hamid et al. 2018).

Statistical analysis:

The statistical analysis were used to analysis the obtained results by SPSS 26 program to analysis of variance (ANOVA) and all means compared using L.S.D at 0.05 (Wagner and William 2015). Some results analyzed by M. Excel (Schmuller, 2013). In addition, statistical treatments were analyzed with probit analysis curve (Finney, 1952).

RESULTS AND DISCUSSION

Mortality tests by green silver nanoparticles on Musca domestica

The presented results in Tables (1 & 2)showed the comparison between sensitive strain and field strains concerning the mortality rate of Musca domestica larvae when they were treated by feeding on the mixed diet with different tested concentrate of Ag NPs 10, 20, 30, 40, and 50 . The mortality % were 47, 57, 73, 90 and 97% susceptibly in sensitive strain after 72 h (Fig., 1) while the field strain recorded a lower percentages than that of the susceptible one 43, 50, 67, 83 and 93% respectively (Fig. 2). The control treatment showed no incidence of larval mortality.

The average larval mortality data were submitted to probit analysis as shown in (Fig. 3 & 4) to determine the LC₅₀ value which calculated by 21.18 , 18.014  in felid and sensitive strains. From this result, the percentage of larval mortality of house fly, M. domestica increased as the concentrations of Ag NPs increased and as the exposure period increased in both strains. There was a significant difference in LC₅₀ values between field strain and sensitive strain represented by a linear relationship. The observed minimal selection effects were caused by developed insecticides resistance in field strain than sensitive one. Scott et al. (1991) suggested that resistance to certain active ingredients occurs among field populations. In addition, synthesized nanoparticles are toxic to the nervous system which more sensitive in susceptible strain than field one Fouad et al. (2018).

Marimuthu et al. (2011) found that, biosynthesized Ag NPs were effective against Aedes aegypti, Culex quinquefasciatus and Anopheles stephensi. In addition, Ag NPs can cause toxicity through a variety of mechanisms, including surface attachment and changes in membrane properties, affect cell permeability (Morones et al., 2005). The physical and chemical properties of Ag NPs can be altered by interactions with biochemical and physiological structures and processes (Nel et al., 2006 and Kim et al., 2009). Insect death can be due to gastrointestinal dysfunction, changes in the skin surface due to dehydration, or obstruction of the stomata and trachea (Abo-Arab et al., 2014).

Table (1) Effects of silver nanoparticles Ag NPs on mortality of susceptible strain 3^rd larval instar larvae after 72h.

Significant at P50 lethal concentration that kills 50% of the exposed larvae UCL upper confidence limit, LCL lower confidence limit, χ2 chi-square, df degree of freedom. A Mean calculated from three replicate.

Table (2) Effects of silver nanoparticles Ag NPs on mortality of field strain 3^rd larval instar of after 72 h.

Significant at P50 lethal concentration that kills 50% of the exposed larvae UCL upper confidence limit, LCL lower confidence limit, χ2 chi-square, df degree of freedom. A Mean  calculated from three replicate.

Fig (1): effects of silver nanoparticles Ag NPs on the mortality of sensitive 3^rd larval instar after 72h.

Fig. (2): effects of silver nanoparticles Ag NPs on the mortality of field strain 3^rd larval instar after 72h

Fig. (3): The probit analysis of the silver nanoparticles Ag NPs on field strain illustrate the liner formula of LC₅₀

Fig. (4): The probit analysis of the silver nanoparticles Ag NPs on sensitive strain illustrate the liner formula of LC₅₀

The pupae weights in the two strains after treatments

The results of the effect of Ag NPs on pupal weight of those larvae survived treatment and formed pupa in table (3) and the results showed that treatments had a significant effect on pupal weights of the two tested strain as they were compared with the control (Fig., 5). It was found the weights of the pupae decreased throughout the pupal stage. The treatment with Ag NPs had a noticeable effect showing the presence of abnormalities in pupae inhibiting adult emergence. Advanced physiological will be test and clear in the next research.

Table (3) the mean of the pupa weight (mg) in field and sensitive strain of M.domestica under effect of silver nanoparticles Ag NPs concentration

Fig (3) the mean of the pupa weight (mg) in field and sensitive strain of M.domestica under effect of silver nanoparticles Ag NPs concentration

Effects of Silver nanoparticles on the two strains adults and their sex ratio

The results in table (4) described the reduction in adult emergence of M. domestica as a consequence effect of the treatment with the green synthesized AgNPs The results showed that the  adults  insects emergence from pupae was affected in field and sensitive strains, and most effective concentration was 50  where the inhibition rate reached 100% , On the other hand, it was noted  effective of Ag NPs on the sex ratio  of adults emergence produced from treated larvae and the efficiency of females  to laying egg ,this was confirmed by the recording of a number of females with deformed ovipositor. These results agree with Abd El-Hamid et al. (2018) where they notice the abnormalities appearance in pupae, and adult stages. In pupae, morphological changes included elongation, dwarfism, and swelling, whereas in adults, some individuals have shown partial emergence and others popped up with wrinkled wings.

Table (4): The effect of silver nanoparticles Ag NPs concentration on adults, their sex ratio and eggs laid in the two strains F & S M domestica

F: field strain S: sensitive strain R.: replicates m.: male f.: female

الجزيئات المتناهةالصغرللفضة الخضراءعلى سلالاتيالذبابةالمنزليةالمقاومةوالحساسة

نادية محمد خميس حسونة¹،أحمد عبد الفتاح محمود عبد المجيد² ،فوز عيسي سعيد عقيلة³ ،فدوي طه محمدين⁴

1 الحشرات الاقتصادية- قسم وقاية النبات  - كلية الزراعة سابا باشا - جامعة الأسكندرية- مصر.

2كمياء وسمية المبيدات – قسم وقاية النبات- كلية الزراعة سابا باشا - جامعة الأسكندرية- مصر.

3 الحشرات الاقتصادية – قسم الحيوان – كلية الزراعة بالبيضا – جامعة عمر المختار- ليبيا.