In vitro evaluation of antileishmanial activity of Boswellia serrata essential oil nanoliposome.

BACKGROUND: Leishmaniasis poses a significant health risk. OBJECTIVES: This study aimed to evaluate the effects of Boswellia serrata (B. serrata) essential oil nanoliposomes on Leishmania tropica (L. tropica) in vitro. METHODS: A mixture of B. serrata essential oil, phosphatidylcholine and Tween 80 were used to prepare B. serrata essential oil nanoliposomes, followed by drying, hydration and size characterisation. The promastigotes of L. tropica were cultured in Roswell Park Memorial Institute medium (RPMI-1640) containing streptomycin, penicillin and fetal bovine serum. Different concentrations of B. serrata essential and nanoliposomes were tested for their antileishmanial properties by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide tests (MTT). RESULTS: Results of Dynamic Light Scattering (DLS) for B. serrata nanoliposomes indicate that they are successful at producing nanoliposomes with dimensions of 74.8 nm. At 1 µg/mL dose, B. serrata essence caused 17 ± 1.73% mortality, while B. serrata nanoliposomes induced 26 ± 1.15% mortality. B. serrata essence achieved a mortality of 55 ± 2.88% at 10 µg/mL, whereas B. serrata nanoliposomes demonstrated a mortality of 63.66±0.88% at 10 µg/mL. Furthermore, there was a significant difference between similar concentrations of B. serrata and B. serrata nanoliposomes. The LC50 of B. serrata essential oil is 7.26 µg/mL in the 95% confidence interval (12.13-5.25). The LC90 value of B. serrata essential oil is 129.37 µg/mL in the 95% confidence interval (50.07-852.58). The LC50 value of B. serrata nanoliposome is 4.20 µg/mL in the 95% confidence interval (6.13-3.10). LC90 value for B. serrata nanoliposome is calculated as 91.89 µg/mL in the 95% confidence interval (37.09-583.29). CONCLUSIONS: In vitro experiments have shown that B. serrata oil and the nanoliposome suppress the proliferation of L. tropica promastigotes, which suggests it may be a promising option for treating leishmaniasis.

Dietary Supplementation with Eugenol Nanoemulsion Alleviates the Negative Effects of Experimental Coccidiosis on Broiler Chicken's Health and Growth Performance.

The present study investigated the protective efficacy of dietary supplementation with clove essential oil (CEO), its main constituent eugenol (EUG), and their nanoformulated emulsions (Nano-CEO and Nano-EUG) against experimental coccidiosis in broiler chickens. To this aim, various parameters (oocyst number per gram of excreta (OPG), daily weight gain (DWG), daily feed intake (DFI), feed conversion ratio (FCR), serum concentrations of total proteins (TP), albumin (ALB), globulins (GLB), triglycerides (TG), cholesterol (CHO) and glucose (GLU), serum activity of superoxide dismutase (SOD), glutathione s-transferase (GST), and glutathione peroxidase (GPx)] were compared among groups receiving CEO supplemented feed (CEO), Nano-CEO supplemented feed (Nano-CEO), EUG supplemented feed (EUG), Nano-EUG supplemented feed (Nano-EUG), diclazuril supplemented feed (standard treatment, ST), or basal diet [diseased control (d-CON) and healthy control (h-CON)), from days 1-42. Chickens of all groups, except h-CON, were challenged with mixed Eimeria spp. at 14 days of age. Coccidiosis development in d-CON was associated with impaired productivity (lower DWG and higher DFI and FCR relative to h-CON; p < 0.05) and altered serum biochemistry (decreased TP, ALB, and GLB concentrations and SOD, GST, and GPx activities relative to h-CON; p < 0.05). ST effectively controlled coccidiosis infection by significantly decreasing OPG values compared with d-CON (p < 0.05) and maintaining zootechnical and serum biochemical parameters at levels close to (DWG, FCR; p < 0.05) or not different from (DFI, TP, ALB, GLB, SOD, GST, and GPx) those of h-CON. Among the phytogenic supplemented (PS) groups, all showed decreased OPG values compared with d-CON (p < 0.05), with the lowest value being measured in Nano-EUG. All PS groups showed better values of DFI and FCR than d-CON (p < 0.05), but only in Nano-EUG were these parameters, along with DWG, not different from those of ST. Furthermore, Nano-EUG was the only PS group having all serum biochemical values not different (or even slightly improved) relative to ST and h-CON. In conclusion, the tested PS diets, especially Nano-EUG, can limit the deleterious effects of coccidiosis in broiler chickens, due to anticoccidial activity and possibly their reported antioxidant and anti-inflammatory properties, thereby representing a potential green alternative to synthetic anticoccidials.

Ameliorative effect of Psidium guajava essential oil supplemented feed on chicken experimental coccidiosis.

Present study investigated the therapeutic efficacy of Psidium guajava essential oil (EO) on chicken experimental coccidiosis in comparison to the diclazuril. Seventy-five 1-day-old Ross 308 broiler chickens were allocated into 5 groups: CEO1: received EO at 1 mg kg-1 of feed; CEO5: received EO at 5 mg kg-1 of feed; CT: received diclazuril as standard treatment; CNT: received only basal diet; NC: control chickens; all of the groups except NC were challenged with mixed Eimeria spp. on d14 and received supplemented diet from d1 to d42. Zootechnical records and oocyst per gram (OPG) of feces samples were analyzed on weekly basis. On the last day of the study, blood samples were taken to measure serum concentrations of biochemical parameters and also activities of antioxidant enzymes. ß-caryophyllene and α-pinene were determined as major constituents of the EO. On the 3rd, 4th, and 5th weeks, a significant difference was noted in feed conversion ratio (FCR) between CEO1, CEO5, and CT in comparison to NC and CNT chickens (p < 0.05). No significant difference was observed in OPG between CEO1 and CEO5 (p > 0.05); however, CT showed a lower number of OPG relative to EO supplemented groups (p < 0.05). The highest serum activity of glutathione peroxidase was observed in CEO5 which was higher than other groups (p < 0.05). Results of the present study showed that supplementation of P. guajava EO especially at 5 mg kg-1 of feed may have beneficial effects on prevention of coccidiosis and improvement of health in broilers.

Acaricidal activity, mode of action, and persistent efficacy of selected essential oils on the poultry red mite (Dermanyssus gallinae).

In this work, the essential oils (EOs) from Litchi chinensis, Clausena anisata, Heracleum sphondylium, Pimpinella anisum, Lippia alba, Crithmum maritimum and Syzygium aromaticum were tested for their contact toxicity against the poultry red mite, Dermanyssus gallinae, a deleterious ectoparasite of aviary systems. In addition, in order to give insights on their mode of action and effectiveness, the vapor phase and residual toxicity tests were also performed. Results showed that amongst all the tested EOs, that of S. aromaticum demonstrated the highest contact toxicity, with a LC50 value of 8.9 µg/mL, followed by C. maritimum and L. chinensis EOs, with LC50 values of 23.7 and 24.7 µg/mL, respectively. L. chinensis and C. anisata EOs showed higher vapor toxicity than the other EOs. L. chinensis and S. aromaticum EOs showed promising toxic effects up to 4 days post-application. Taken together, these results highlighted L. chinensis and S. aromaticum as two promising sources of biopesticides, able to cause severe contact, vapor and residual toxicity in the poultry red mites. Given the wide plant cultivation and uses in foodstuffs, cosmetics, flavour and fragrances, these EOs may be considered cheap and ready-to-use products as valid, eco-friendly alternatives to pesticides currently used in the aviary systems.

Towards green drugs against cestodes: Effectiveness of Pelargonium roseum and Ferula gummosa essential oils and their main component on Echinococcus granulosus protoscoleces.

Cystic echinococcosis is of great public health importance. Surgery is the efficient treatment for this infection. To minimize the risk of cyst intraoperative leakage, using scolicidals is crucial. To date, any efficacious scolicidal without side effect has not been introduced. Since essential oils of Pelargonium roseum and Ferula gummosa have shown several bioactivities, we evaluated their potential against protoscoleces of E. granulosus using albendazole as positive control. Furthermore, chemical composition of both essential oils was analyzed by gas chromatography-mass spectrometry (GC-MS) analyses, and their main constituents were also evaluated for scolicidal activity. Different concentrations of essential oils and their two main constituents were tested for scolicidal activity. Mortality rate was measured by eosin staining. Results of GC-MS revealed citronellol and ß-pinene as the main constituents of P. roseum and F. gummosa essential oils, respectively. After 60 min of exposure to 50 µg/mL of P. roseum and F. gummosa, mean mortality rate of protoscoleces was 100%. However, ß-pinene and citronellol at the same time point with only 10 µg/mL concentrations resulted in approximately higher than 80% mortality. Essential oils of P. roseum and F. gummosa showed significant toxic effect on E. granulosus with 50% lethal concentration (LC50) values of 8.52 and 17.18 µg/mL, respectively. Based on the LC50 values, ß-pinene (2.20 µg/mL) was the most potent scolicidal agent in the present study. The overall toxicity of ß-pinene and citronellol was significantly higher than the whole essential oils of F. gummosa and P. roseum. Based on these results, ß-pinene and citronellol can be considered as candidate ingredients for the development of green scolicidals.

Toxic and repellent activity of selected monoterpenoids (thymol, carvacrol and linalool) against the castor bean tick, Ixodes ricinus (Acari: Ixodidae).

The castor bean tick, Ixodes ricinus, is a species of medical and veterinary importance. The use of synthetic acaricides for tick control has led to development of resistance, residues in the environment and animal products, and public health concerns. In this regard, plant essential oils and their main constituents represent an appealing alternative strategy to combat ticks. The phenols thymol and carvacrol and the alcohol linalool are monoterpenoids occurring in essential oils of several aromatic and medicinal plants, such as thyme, oregano, savory, lavender and coriander. Recent studies have shown toxicity of these monoterpenoids against selected mosquito vectors and other arthropod pests. However, information on their bioactivity on I. ricinus is not available. On this basis, here we evaluated the ovicidal, larvicidal and repellency effects of these compounds against I. ricinus. Concentrations of 0.25, 0.5, 1, 2 and 5% were sprayed on the egg masses, then hatching rates were noted. Larvicidal assays were conducted on unengorged larvae, following the larval packet technique. The repellency was determined by measuring the vertical migration behavior of ticks in laboratory conditions. Carvacrol and thymol at all concentrations tested led to a significant hatching decrease, showing an efficacy higher than permethrin, whereas linalool did not cause any significant effect. In the larvae treated with carvacrol and thymol (1, 2 and 5%), mortality rates reached 100% after 24h, showing a larvicidal efficacy higher than permethrin, whereas no effect was seen in the larval groups treated with linalool. Carvacrol and thymol at all concentrations tested showed >90% repellency on I. ricinus. Linalool was scarcely effective (50.24% repellency) only at the concentration of 5%. Overall, based on these results, the phenols carvacrol and thymol can be considered as candidate ingredients for the development of novel acaricidal formulations to control the populations of I. ricinus and the spread of related tick-borne diseases.

Eco-friendly control of the poultry red mite, Dermanyssus gallinae (Dermanyssidae), using the α-thujone-rich essential oil of Artemisia sieberi (Asteraceae): toxic and repellent potential.

The poultry red mite, Dermanyssus gallinae, represents a key threat for the poultry industry worldwide. The control of D. gallinae is mainly achieved by continuous applications of acaricides. However, the fast-growing development of resistance, and the strict laws concerning chemicals admitted for treatments on food animals, highlighted the importance of alternative control tools. Here, we explored the potential of Artemisia sieberi essential oil against D. gallinae. In this study, the A. sieberi essential oil was analyzed using GC and GC-MS. The oil toxicity through contact and fumigant assays on adult mites was evaluated. The oil repellent activity was assessed on adult mites over different time intervals. Lastly, the residual toxicity of various doses of the oil was evaluated on D. gallinae until 14 days post treatment. GC and GC-MS showed that the oil was rich in α-thujone (31.5%), ß-thujone (11.92%), camphor (12.3%), and 1,8-cineole (10.09%). Contact toxicity on adult mites showed 50% lethal concentration (LC50), LC90, and LC99 of 15.85, 26.63, and 35.42 µg/cm3, respectively. In fumigant assays, the oil was toxic on D. gallinae, and mortality was significantly higher in open containers over closed ones, underlining the key role of highly volatile constituents. Repellent assays showed that after 24 h from the treatment, all doses of the A. sieberi essential oil led to significant repellent activity over the control, except for 2 µg/cm3. After 48 h, A. sieberi essential oil tested at all doses led to significant repellent activity, if compared to the control. Residual toxicity assays showed that time exposure and concentration tested had a significant impact on mite mortality after 1, 2, 5, and 7 days from the treatment. Notably, mortality remained significantly higher over the control for 7 days after spraying with oil at 2%. Further field assays with selected molecules from the A. sieberi essential oil are ongoing, testing them in synergistic blends, as well as in microencapsulated formulations.

Toxicity of ß-citronellol, geraniol and linalool from Pelargonium roseum essential oil against the West Nile and filariasis vector Culex pipiens (Diptera: Culicidae).

Insect vectors are responsible for spreading devastating parasites and pathogens. A large number of botanicals have been suggested for eco-friendly control programs against mosquito vectors, and some of them are aromatic plants. Pelargonium roseum, a species belonging to the Geraniaceae family, due to its pleasant rose-like odor may represent a suitable candidate as mosquito repellent and/or larvicide. In this research, we evaluated the toxicity of the essential oil from P. roseum and its major constituents against the West Nile and filariasis vector Culex pipiens. The chemical composition of P. roseum essential oil was analyzed by gas chromatography-mass spectroscopy. Major constituents were citronellol (35.9%), geraniol (18.5%), and linalool (5.72%). The bioactivity of P. roseum essential oil and its three major compounds on larvae and egg rafts of Cx. pipiens was evaluated. The essential oil had a significant toxic effect on larvae and egg rafts of Cx. pipiens, with 50% lethal concentration (LC50) values of 5.49 and 0.45µg/mL, respectively. Major constituents, geraniol, citronellol and linalool resulted in LC50 values of 6.86, 7.64 and 14.87µg/mL on larvae, and 0.8, 0.67 and 1.27µg/mL on egg rafts. Essential oil and two of its constituents, citronellol and geraniol showed moderate knock-down on Cx. pipiens adults. Overall, the present investigation revealed that the major components of P. roseum and specially the whole essential oil could be helpful in developing novel and safe mosquito control tools and also offer an environmentally safe and cheap tool for reducing Cx. pipiens mosquito populations.

Traps containing carvacrol, a biological approach for the control of Dermanyssus gallinae.

Resistance to conventional synthetic pesticides has been widely reported in Dermanyssus gallinae in different aviary systems. Cardboard traps containing acaricides had been introduced as a successive device for collection and control of the poultry red mite. The present study assessed field efficacy of traps containing carvacrol in the control and reduction of D. gallinae in laying poultry farm. Two different carvacrol-based formulations were tested for their toxicity and possible repellent activity on D. gallinae to determine the most appropriate formulation and concentration to be used in the field study. In vitro tests confirmed that 1 % carvacrol formulation with ethoxylated castor oil as emulsifier was significantly toxic to D. gallinae without any dissuading effect in comparison to ethanol and higher concentrations of carvacrol (p < 0.05). A subsequent in vivo experiment in a cage system laying farm demonstrated significant acaricidal activity for traps containing 1 % carvacrol. Throughout the study, untreated cardboard traps were used for monitoring mite populations. Carvacrol-impregnated traps were efficacious in the control of D. gallinae and led to over 92 % reduction in mite's population after 2 week of application. Toxic effects of carvacrol maintained through 2 weeks after the last application of traps. Results of the present study suggested that effective control of the poultry red mite can be achieved by traps containing carvacrol. These traps can be used safely in poultry facilities without any concern about residues in eggs, meat, and environment.

Survey on efficacy of chloroformic extract of Artemisia annua against Giardia lamblia trophozoite and cyst in vitro.

Giardiasis is a parasitic cosmopolitan disease that the rate of infection in developing countries is considerable. This infection directly is associated with poor hygienic conditions, poor water quality control, and overcrowding. Reinfection and drug resistance are two major problems in endemic areas. Recently, researchers are concentrating on herbal drugs as a proper solution. Therefore, the objective of the present study was to survey on efficacy of chloroformic extract of Artemisia annua against Giardia lamblia trophozoite and cyst in vitro. G. lamblia cysts were prepared from faces of giardiasis patients from different hospitals of Mazandaran Medical University. Four concentrations (1, 10, 50 and 100 mg/ml) of chloroformic extract of A. annua were utilized for 1, 5, 30, 60 and 180 min. Viability of G. lamblia cysts was confirmed by 0.1 % Eosin staining. Cyst and trophozoite contact (intermix) of G. lamblia with extract of A. annua with variant concentrations (1, 10, 50 and 100 mg/ml) after 1 and 180 min caused following cyst and trophozoite elimination rates: (67, 69, 71 and 73 %), (65, 67, 67 and 72 %), (94, 96, 97 and 99 %) and (100, 100, 100 and 100 %), respectively. Authors from the current investigation draw a conclusion that chloroformic extract of A. annua has the ability to eliminate G. lamblia cysts and trophozoites in vitro.

Synergistic effect of aged garlic extract and naltrexone on improving immune responses to experimentally induced fibrosarcoma tumor in BALB/c mice.

BACKGROUND: Garlic, a medicinal plant, and Naltrexone (NTX), an opioid receptor antagonist, both have immunomodulatory and antitumor effects. Current study was designed to evaluate synergistic antitumor effects of aged garlic extract (AGE) and NTX. MATERIALS AND METHODS: WEHI-164 fibrosarcoma cells were implanted subcutaneously on day 0 into right flank of 80 BALB/c mice at age of 8 weeks. Mice were randomly categorized in four separate groups: The first group received AGE (100 mg/kg, i.p.), the second group received NTX (0.5 mg/kg, i.p.), the third group received both of them, and the fourth group received phosphate buffered saline as control group. Treatments were administered three times per week. Tumor growth was measured and morbidity was recorded. Subpopulations of CD4+/CD8+ T cells were determined using flowcytometery. WEHI-164 cell specific cytotoxicity of splenocytes and in vitro production of interferon-gamma (IFN-γ) and interleukin-4 (IL-4) cytokines were measured. All statistical analyses were conducted with SPSS 16 software and P < 0.05 was considered to be statistically significant. RESULTS: The mice who received AGE+NTX had significantly longer survival time compared with the mice treated with AGE or NTX alone. An enhanced inhibitory effect on tumor growth was seen in combination therapy group. The CD4+/CD8+ ratio and in vitro IFN-γ production of splenocytes were significantly increased in AGE+NTX and NTX groups. WEHI-164 specific cytotoxicity of splenocytes was also significantly increased at 25:1 E:T ratio in AGE+NTX treated mice. Coadministration of AGE with NTX resulted in improvement of immune responses against experimentally implanted fibrosarcoma tumors in BALB/c mice. CONCLUSIONS: AGE showed synergistic effects with NTX on inhibition of tumor growth and increment of survival times.