Acaricidal activity of Artemisia herba-alba and Melia azedarach oil nanoemulsion against Hyalomma dromedarii and their toxicity on Swiss albino mice.

Biopesticides such as essential oils (EOs) are considered an improvement for integrated pest control as they appear to be less toxic to the environment than chemical acaricides. The current study aimed to evaluate the acaricidal activity of Artemisia herba-alba and Melia azedarach oil loaded nano-emulsion as alternatives for chemical acaricides against the camel tick Hyalomma dromedarii, besides evaluating their toxic effect on Swiss albino mice. Transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR) were used for the characterization of loaded nano-emulsions.The immersion test was used for the bioassay of both loaded nanoemulsions on tick stages (egg, nymph, larva, and adult). Mortality percentages and LC50 values of each tick stage were calculated. Reproductive performance for the survived engorged females after treatment was monitored. The toxicity of both loaded nano-emulsions was evaluated on Swiss albino mice by an oral dose of 1500 mg/kg/day for five successive days. The hematological, biochemical, and histopathological changes were evaluated. TEM characterization revealed spherical droplets for A. herba-alba and M. azedarach oil loaded nano-emulsion with droplet size ranging from 62 to 69 nm and 52-91 nm, respectively. FTIR revealed the absence of extra peaks in the loaded nano-emulsions that confirmed no chemical changes existed by ultrasonication. The LC50 values of A. herba-alba and M. azedarach oil loaded nano-emulsion on embryonated eggs, larvae, engorged nymphs, and unfed adults were 0.3 and 1.1%, 0.7 and 1.7%, 0.3 and 0.4%, 4.4 and 22.2%, respectively. The egg productive index (EPI), egg number, and hatchability percentage were lower in the treated females compared with Butox 5% (deltamethrin) and control. The hematological picture and biochemical analysis revealed insignificant changes in the treatment group compared with the negative control group. The liver of the A. herba-alba and M. azedarach oil loaded nano-emulsion treated group exhibited vacuolar degeneration and infiltration of lymphocytic cells. The kidney of mice treated with A. herba-alba and M. azedarach oil loaded nano-emulsion showed hemolysis and slight degeneration of epithelial cells of tubules. It is concluded that A. herba-alba and M. azedarach oil loaded nano-emulsion have good acaricidal activity against camel tick H. dromedarii.

Therapeutic Potential of Green Synthesized Copper Nanoparticles Alone or Combined with Meglumine Antimoniate (Glucantime®) in Cutaneous Leishmaniasis.

BACKGROUND: In recent years, the focus on nanotechnological methods in medicine, especially in the treatment of microbial infections, has increased rapidly. AIM: The present study aims to evaluate in vitro and in vivo antileishmanial effects of copper nanoparticles (CuNPs) green synthesized by Capparis spinosa fruit extract alone and combined with meglumine antimoniate (MA). METHODS: CuNPs were green synthesized by C. spinosa methanolic extract. The in vitro antileishmanial activity of CuNPs (10-200 µg/mL) or MA alone (10-200 µg/mL), and various concentrations of MA (10-200 µg/mL) along with 20 µg/mL of CuNPs, was assessed against the Leishmania major (MRHO/IR/75/ER) amastigote forms and, then tested on cutaneous leishmaniasis induced in male BALB/c mice by L. major. Moreover, infectivity rate, nitric oxide (NO) production, and cytotoxic effects of CuNPs on J774-A1 cells were evaluated. RESULTS: Scanning electron microscopy showed that the particle size of CuNPs was 17 to 41 nm. The results demonstrated that CuNPs, especially combined with MA, significantly (p < 0.001) inhibited the growth rate of L. major amastigotes and triggered the production of NO (p < 0.05) in a dose-dependent manner. CuNPs also had no significant cytotoxicity in J774 cells. The mean number of parasites was significantly (p < 0.05) reduced in the infected mice treated with CuNPs, especially combined with MA in a dose-dependent response. The mean diameter of the lesions decreased by 43 and 58 mm after the treatment with concentrations of 100 and 200 mg/mL of CuNPs, respectively. CONCLUSION: The findings of the present study demonstrated the high potency and synergistic effect of CuNPs alone and combined with MA in inhibiting the growth of amastigote forms of L. major, as well as recovery and improving cutaneous leishmaniasis (CL) induced by L. major in BALB/c mice. Additionally, supplementary studies, especially in clinical settings, are required.

In vitro acaricidal activity of green synthesized nickel oxide nanoparticles against the camel tick, Hyalomma dromedarii (Ixodidae), and its toxicity on Swiss albino mice.

The green synthesized nanoparticles have been determined as a novel pesticide against arthropod pests. This study was designed to evaluate the in vitro acaricidal activity of green synthesized nickel oxide nanoparticles (NiO NPs) using aqueous extract of Melia azedarach ripened fruits against different developmental stages of the camel tick Hyalomma dromedarii in addition to their toxic effect on laboratory animals. The synthesized NiO NPs were characterized by UV-visible (UV-Vis) spectroscopy, Fourier transforms infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDS). The UV-Vis spectra of the NiO NPs showed an absorption peak at 307 nm. FTIR analysis showed the possible functional groups used for capping and stabilization of NiO NPs with strong bands at 3416.2 and 1626.6 cm-1. The SEM images of the NiO NPs exhibited a size ranging from 21 to 35 nm. The immersion test was used for the in vitro application of the synthesized NiO NPs on the various tick stages (egg, nymph, larva, and adult). Mortality percentages and LC50 values of each tick stage were calculated. The oviposition and hatchability of the engorged females were monitored for the survived tick after treatment. The LC50 values for NiO NPs on embryonated eggs, larvae, and engorged nymphs were 5.00, 7.15, and 1.90 mg/mL, respectively. The egg productive index (EPI), egg number, and hatchability (%) were lower in females treated with the NiO NPs than in control ticks. The toxicity of the NiO NPs on laboratory animals was also investigated using Swiss albino mice by oral dose of 500 mg/kg/day administration for five consecutive days. The hematological, biochemical, and histopathological changes were evaluated. The hematological analysis showed significant increase in the level of white blood cells (WBC) and hemoglobin (Hb). Biochemical analysis showed non-significant decrease in alkaline phosphatase (ALP) and alanine amino transferase (ALT). We concluded that NiO NPs have a significant acaricidal activity as demonstrated on eggs, larvae, engorged nymphs, and fully fed females of H. dromedarii. From a toxicological point of view further in vivo investigations are needed to determine the mechanism of toxic effect of NiO NPs.

Myrtus communis Essential Oil; Anti-Parasitic Effects and Induction of the Innate Immune System in Mice with Toxoplasma gondii Infection.

BACKGROUND: Myrtus communis (M. communis) is a wild aromatic plant used for traditional herbal medicine that can be demonstrated in insecticidal, antioxidant, anti-inflammatory, and antimicrobial activity of its essential oils (MCEO). AIM: The present study aimed to evaluate the prophylactic effects of M. communis essential oil (MCEO) against chronic toxoplasmosis induced by the Tehran strain of Toxoplasma gondii in mice. METHODS: Gas chromatography/mass spectrometry (GC/MS) analysis was performed to determine the chemical composition of MCEO. Mice were then orally administrated with MCEO at the doses of 100, 200, and 300 mg/kg/day and also atovaquone 100 mg/kg for 21 days. On the 15th day, the mice were infected with the intraperitoneal inoculation of 20-25 tissue cysts from the Tehran strain of T. gondii. The mean numbers of brain tissue cysts and the mRNA levels of IL-12 and IFN-γ in mice of each tested group were measured. RESULTS: By GC/MS, the major constituents were α-pinene (24.7%), 1,8-cineole (19.6%), and linalool (12.6%), respectively. The results demonstrated that the mean number of T. gondii tissue cysts in experimental groups Ex1 (p < 0.05), Ex2 (p < 0.001) and Ex3 (p < 0.001) was meaningfully reduced in a dose-dependent manner compared with the control group (C2). The mean diameter of tissue cyst was significantly reduced in mice of the experimental groups Ex2 (p < 0.01) and Ex3 (p < 0.001). The results demonstrated that although the mRNA levels of IFN-γ and IL-12 were elevated in all mice of experimental groups, a significant increase (p < 0.001) was observed in tested groups of Ex2 and Ex3 when compared with control groups. CONCLUSION: The findings of the present study demonstrated the potent prophylactic effects of MCEO especially in the doses 200 and 300 mg/kg in mice infected with T. gondii. Although the exceptional anti-Toxoplasma effects of MCEO and other possessions, such as improved innate immunity and low toxicity are positive topics, there is, however, a need for more proof from investigations in this field.

Ovicidal and Latent Effects of Pulicaria jaubertii (Asteraceae) Leaf Extracts on Aedes aegypti.

The control of Aedes aegypti with synthetic pesticides may result in adverse effects on wildlife and the environment. Bioactive plant extracts have been proposed as one of the alternatives to chemical pesticides used against mosquitoes. Here, we report on the ovicidal and latent effects of ethanolic, petroleum ether, and chloroform leaf extracts of Pulicaria jaubertii at 25 to 150 ppm each against the life stages of laboratory stain of Ae. aegypti. At 150 ppm, the ethanolic leaf extract resulted in 100% ovicidal activity, followed by petroleum ether extract (74%), and chloroform extract about 7% mortality. The ethanolic extract produced 100% larval and pupal mortality at both 75 and 50 ppm, while the petroleum ether extract produced 76.5 and 58.3%, respectively. The ethanolic extract recorded the highest percentage of adult mortality (72.7%) at the lowest concentration (25 ppm). At 25 and 50 ppm, the ethanolic extract resulted in 62.2 and 85.2% sterility index of Ae. aegypti females, respectively, as compared with the 0.1 and 3.5% sterility index caused by the chloroform extract at the same concentrations. In conclusion, P. jaubertii appears to have potential to be further evaluated as a mosquito control agent. Additional studies are needed on its mode of action, synergism with other products, and efficacy under actual field conditions.

In vitro acaricidal effect of Melia azedarach and Artemisia herba-alba extracts on Hyalomma dromedarii (Acari: Ixodidae): embryonated eggs and engorged nymphs.

The present study aimed to evaluate the in vitro efficacy of four medicinal plant extracts: petroleum ether and ethyl alcohol extracts of the ripen fruits of Melia azedarach and whole aerial parts of Artemisia herba-alba against the two inactive stages of the camel tick Hyalomma dromedarii, embryonated eggs and engorged nymphs in comparison to reference acaricide Butox®5.0 (Deltamethrin). Egg and nymphal immersion tests at four concentrations with three replicates were used. The deformity in larvae hatched from treated eggs and adults moulted from treated nymphs were observed and photographed by light microscope (LM) and scanning electron microscope (SEM). The results showed that M. azedarach and A. herba-alba extracts revealed higher significant toxic effects on embryonated eggs and engorged nymphs comparing with the reference acaricide (Butox®5.0) and control. In egg emmersion test, the LC50 of petroleum ether extracts of M. azedarach and A. herba-alba was 3.14 and 3.91%, respectively and LC50 of the respective ethyl alcohol extracts was 1.77 and 2.45%. In nymphal immersion test, LC50 of petroleum ether extracts of M. azedarach and A. herba-alba was 0.26 and 1%, respectively, and LC50 of the respective ethyl alcohol extracts was 4.17 and 8.7%. Abnormalities were observed by LM and SEM in the larvae hatched from the treated eggs as incomplete development of legs and mouth parts as well as shrinkage mainly in legs and mouthparts of adults emerged from treated nymphs. In conclusion, all extracts and petroleum ether extracts of the two plants have great potential to be developed as a novel acaricidal for controlling eggs and nymphs of H. dromedarii, respectively.

Efficacy and safety of ethanolic Curcuma longa extract as a treatment for sand tampan ticks in a rabbit model.

BACKGROUND AND AIM: The soft tick Ornithodoros savignyi is distributed throughout Africa, including Egypt. It primarily attacks camels, cattle, donkeys, and cows; and rarely affects humans. This study evaluated the acaricidal efficacy of ethanolic Curcuma longa extract (Turmeric) on the second nymphs of O. savignyi and then investigated the safety of this herb in rabbits. MATERIALS AND METHODS: The nymphs were immersed in 10, 5, 2.5, 1.25, and 0.625 mg/ml ethanolic C. longa extract. An additional group was immersed in ethanol as a control. On the 1st, 7th, and 15th-day post-treatment, the mortality percentages, LC50, and LC95 were calculated. The ticks exposed to 10mg/ml ethanol C. longa extract were investigated by scanning electron microscopy (SEM). Three male New Zealand White rabbits were orally administered 2ml (two doses) of 10mg/ml ethanolic C. longa extract, and another three rabbits were orally given two doses of 2ml of absolute ethanol as a negative control. Histopathological examination of the kidney and liver hematology and the kidney and liver function was performed. Chemical analysis of the extract was determined by gas chromatography-mass spectrometry (GC-MS). RESULTS: The LC50 and LC95 were 1.31 and 15.07, 1.07 and 8.56, and 0.81 and 6.97mg/ml on the 1st, 7th, and 15thday, respectively. SEM revealed that mamillae and spots on the surfaces of the treated ticks were not discriminating except for some clefts on the surfaces. The histological examination, blood profile, and biochemical analyses revealed no significant differences between the treated and untreated rabbits (p>0.05). GC/MS analysis revealed 50 compounds, and curcumene and tumerone were found to be the major constituents of this ethanolic extract. CONCLUSION: The ethanolic C. longa extract produced a strong acaricidal effect on the second nymph of O. savignyi, and it was safe to use in rabbits.

The efficacy of some wild medicinal plant extracts on the survival and development of third instar larvae of Chrysomyia albiceps (Wied) (Diptera: Calliphoridae).

Four crude wild plants extract of Artemisia herba-alba Asso, Artemisia monosperma Del., Euphorbia aegyptiaca Boiss. and Francoeuria crispa (Forsk.) extracted with four successive solvents; hexane, diethyl ether, ethyl acetate and ethanol were evaluated against the third instar larvae of Chrysomyia albiceps using dipping and thin film techniques. In dipping technique, larvae were immersed in the concentrations of plant extracts for 30 seconds. However, in thin film technique, larvae were exposed to thin layer from each plant extract in the Petri-dishes. Results showed that all extracts had toxic effects on larvae in both two treatments. Hexane and diethyl ether extracts of A. herba-alba and ethyl acetate extract of A. monosperma recorded the highest effect in both two treatments. Ethanol extracts of E. aegyptiaca and A. monosperma were the highest in dipping and thin film treatments, respectively. In dipping treatment, most plant extracts revealed extending effect on pupae especially while in thin film treatment most larvae which succeeded to develop to pupae produced normal flies. Deformed pupae were only recorded with the high concentration of diethyl extracts of A. herba-alba. Hexane, diethyl ether and ethyl acetate extracts of F. crespa only produced low percentages of deformed flies. Histological examination conducted on larvae confirmed that extracts in thin film treatment penetrated to the gut and destroyed its epithelial cells and wall. It concluded that the crude extracts of the four tested plants can be used in controlling of C. albiceps larvae while hexane extracts of E. aegyptiaca, A. herba-alba and A. monosperma are considered the most promising plant preparations against the larvae by using thin film technique.

Acaricidal efficacy of Myrrh (Commiphora molmol) on the fowl tick Argas persicus (Acari: Argasidae).

Five concentrations of purified extract of Myrrh from Commiphora molmol tree were prepared to study its effects on the fowl tick Argas persicus under laboratory conditions. The results revealed that Myrrh had dependant dose toxic effect on the adult female of A. persicus. Toxicity increased gradually daily post treatment. The LC50 was 1.28%, 0.88%, 0.84%, 0.50% and 0.42% at Ist, 2nd, 3rd, 6th and 12th days respectively. At 12th day, the recorded mortality rates were 63, 67, 76, 87 and 94% for concentrations, 0.625, 1.25, 2.5, 5 and 10%, respectively against 5% in control. Histopathological and Transmission election microscope (TEM) examinations showed the lysing of epithelial gut cells in treated groups. The lysed epithelial gut cells showed irregularly distributed nucleus, commonly at low concentrations and rarely in high concentrations of Myrrh. The lysed epithelial gut cells, without nucleus or with aggregated one beside the basal lamina, were common at high concentrations and rare in low concentrations of Myrrh. Consequently, Myrrh can rapidly penetrate the cuticle to body cavity, destroy the epithelial gut cells and finally cause the death of ticks.