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Background: Avian trichomoniasis is an important disease that causes bird mortality, both wild and captive, around the world. This study evaluated the in vitro cytotoxicity, antioxidant, and antiparasitic activity of citral (3.7-Dimetil-2.6-octadienal) and geraniol (trans-3.7-Dimetil-2.6-octadien-1-ol) against Trichomonas gallinae trophozoites. Methods: In vitro assays were conducted at the Laboratory of Protozoology and Entomology (LAPEN) at the Federal University of Pelotas (UFPel), Brazil in 2019 using tests with 106 parasites and citral and geraniol at concentrations ranging from 10 to 100 μM and four controls: NC (culture medium and trophozoites), MTZ (trophozoites plus 100 μM of metronidazole), and TW (trophozoites plus vehicles used for solubilizing derivatives (0.01% Tween). Results: The citral (60 μM) and geraniol (50 μM) concentrations reduced the trophozoitesʼs viability by 100%. The molecular docking experiment demonstrated that citral and geraniol might inhibit a hydrogen enzyme for T. gallinae survival. Conclusion: The major compounds of lemongrass have potential antitrichomonal activity against T. gallinae in vitro.

Background Propolis is a natural resinous product from a variety of plants and combines it with beeswax and salivary enzymes to create bee glue. Its potentials have been employed in the treatment of many diseases and conditions for centuries. This study, therefore, aims to determine the anti-Acanthamoeba properties of the propolis extract, eye drops coupled to some mechanisms such as inhibition of encystation and excystation. Methods The minimum inhibitory concentrations (MICs) of the most active propolis extract were assessed in trophozoites and cysts of Acanthamoeba triangularis (A. triangularis) at 0.256 and 1 mg/mL, respectively. Results Active eye drops inhibited trophozoites and cysts with a MIC value of 100%. At concentrations lower than their MICs values (1/2–1/16 MIC), propolis and eye drops revealed inhibition of encystation. In addition, at 1/2 MIC, both the propolis extract and eye drops showed potential inhibition of excystation. In combined sets of propolis extract and eye drops, they showed slightly increasing inhibition of encystation and excystation. Surprisingly, the MIC of chlorhexidine decreased when combined with the propolis and eye drops. SEM and TEM images displayed morphological changes in A. triangularis treated with combined sets of propolis extract and eye drops. Conclusion From this study, a new combined eye drop solution with propolis extract was found to be helpful in reducing encystation and excystation in A. triangularis. Therefore, this combination is an alternative for the treatment of eye diseases in early stages along with reducing the entry into the cyst stage of Acanthamoeba. The results of this study revealed new anti-Acanthamoeba inhibitors with promising combined alternative therapeutic potential for the treatment of Acanthamoeba keratitis.

Introduction: Echinococcus granulosus is the causative agent of hydatid cysts, which are distributed worldwide. Nowadays, much research is being done to inactivate hydatid cyst protoscoleces. This study was conducted with the aim to compare the scolicidal effect of two chemical compounds 12 and 2 on hydatid cyst protoscoleces in vitro. Material & Methods: The scolicidal activity of two amini-hydrox-phenylcoumarin and amini-bromo-phenylcoumarin at concentrations of 1.2, 1.5, 1.10 and 1.100 after 15, 30 and 60 minutes of incubation was tested separately and each of the experiments was repeated three times. The viability of protoscoleces was checked with 0.1% eosin dye under a light microscope. Subsequently data were analyzed using GraphPad version 5 software and two-way repeated measures ANOVA and values (P > 0.05) was considered a significant level. Findings: The results of this study showed that the chemical compound amini-bromophenylcoumarin at a concentration of 1.10 destroyed 100% of protoscoleces after 60 minutes of exposure compared to the control group, but amini-hydroxophenyl-coumarin, at a concentration of 1.100 could destroy 98% of protoscolecia after 60 minutes of exposure. Discussion & Conclusion: The results of the present study showed that amini-bromophenylcoumarin had stronger protoscolicidal effects than amini-hydroxophenyl-coumarin. However, further studies and in vivo studies are needed to evaluate the efficacy of these chemical compounds.

Background: Nitrite and nitrate salts are used as preservatives in meat products to prevent the growth of pathogen organisms such as Clostridium botulinum. Nitrogen salts additives create the desired color in meat products. However, the use of these salts accelerates nitrosamine production and increases the risk of carcinogenesis. Hence, nitrate and nitrite consumption rate should be according to the approved standards. This analytical study is designed to determine nitrate and nitrite residue of non-heat-treated meat products including hamburger and kebab in East Azerbaijan Province, Iran during 2015-2020. Methods: 120 samples of non-heat-treated meat products from different brands were randomly collected. They consisted of 60 products manufactured in East Azerbaijan province and 60 products imported from other provinces. Nitrate and Nitrite residues in non-heat-treated meat products including hamburger and kebab were determined by spectrophotometric method. Results: Results showed that the residue levels of nitrate and nitrite content regarding non-heat- treated meat products were less than maximum residual limits (MRLs) of national standards. The study also indicated that frozen non-heated meat products did not have any nitrogenous additive. Conclusion: The results of this research revealed that the frozen non-heat treated meat products including hamburger and kebab, produced or consumed in East Azerbaijan province during 2015-2020 did not have nitrate and nitrite additives. Continuous monitoring to ensure the compliance of meat products with the standards is necessary.

Background: Trichomonas vaginalis, a parasitic flagellated protozoan, is one of the main non-viral sexually transmitted diseases worldwide. Treatment options for trichomoniasis are limited to nitroimidazole compounds. However, resistance to these drugs has been reported, which requires the development of new anti-Trichomonas agents that confer suitable efficacy and less toxicity. Methods: In the present work, we assessed the effectiveness of the liposomal system containing essential oils of Bunium persicum and Trachyspermum ammi against T. vaginalis in vitro. The chemical composition of B. persicum and T. ammi were analyzed using gas chromatography-mass spectrometry (GC–MS). Liposomal vesicles were prepared with phosphatidylcholine) 70%) and cholesterol)30%) using the thin-film method. The essential oils of B. persicum and T. ammi were loaded into the liposomes using the inactive loading method. Liposomal vesicles were made for two plants separately. Their physicochemical features were tested using Zeta-Sizer, AFM and SEM. The anti-Trichomonas activity was determined after 12 and 24 h of parasite cultures in TYI-S-33 medium. Results: After 12 and 24 h of administration, the IC50 of the B. persicum essential oil nano-liposomes induced 14.41 μg/mL and 45.19 μg/mL, respectively. The IC50 of T. ammi essential oil nano-liposomes induced 8.08 μg/mL and 25.81 μg/mL, respectively. Conclusions: These data suggested that nano-liposomes of the essential oils of B. persicum and T. ammi may be a promising alternative to current treatments for Trichomonas infection.

Cystic echinococcosis (CE) is a zoonotic disease and with has a global distribution. Today, much research carries out to inactivate hydatid cyst protoscoleces. In particular, herbal compounds have received more attention due to their cheapness, easy access, low toxicity, and side effects. This study aimed to compare the scolicidal effect of hydroalcoholic extract of Allium sativum (garlic) and Ferula asafoetida (angozeh) on hydatid cyst protoscoleces in vitro. The scolicidal activity of A. sativum and F. asafoetida extracts were evaluated at concentrations of 50, 100, 150, 200, and 250 mg/ml following 10, 30, and 60 minutes of exposure. Each reaction was repeated three times. The viability of protoscoleces was examined with a 0.1% eosin stain under a light microscope. The chemical composition of two extracts was analyzed using Gas ChromatographyMass Spectrometry (GC–MS). Statistical analyses were performed with GraphPad software version 5.0. The results of this study showed that F. asafoetida extract, at a concentration of 250 mg/ml after 60 minutes of exposure, killed 100% of the protoscoleces compared to the control group, but the hydroalcoholic extract of A. sativum at the same concentration and time, it was able to kill 98% of protoscoleces. The main chemical components of A. sativum and F. asafoetida identified as allyl methyl trisulfide (12.8%) and methyl ester (13.9%), respectively. The findings of the present study showed that F. asafoetida has more potent scolicidal effects than A. sativum. However, further studies are needed to evaluate the effectiveness of the F. asafoetida plant.

Background and Objectives: Cutaneous leishmaniasis is one of the most important health-threatening diseases in Iran and other countries. Glucantime is currently used to treat this disease, but due to its side effects and high resistance, alternative therapies such as the use of nanoparticles have been considered by researchers. This study aims to investigate the anti-leishmania activity of silver nanoparticles on Leishmania major in vitro. Methods: This is an experimental study on the anti-leishmania activities of silver nanoparticles at different concentrations of 0.75-0.96 µg/ml after 24, 48 and 72 hours of exposure to 106 live Leishmania major promastigotes. The numbers of live parasites were counted by Trypan Blue on a neobar slide using optical microscope (Hemocytometer method). Glucantime and distilled water were considered as positive and negative controls, respectively. The half-maximal inhibitory concentration (IC50) was calculated by SigmaPlot™ software, v‌ersion 13. All reactions were done three times and their average was considered as final result. Results: All concentrations of silver nanoparticles had anti-leishmania activity, where the concentration of 96 µg/ml had the highest effect (100%) 72 hours after exposure. The IC50 was obtained 36.67, 27.2 and 21.08 µg/ml after 24, 48 and 72 hours of exposure, respectively. Conclusion: Silver nanoparticles have an inhibitory effect on the growth of Leishmania major in different concentrations. However, further in-vivo studies are needed to determine the effictivness of silver nanoparticles.

Background : Propolis is a natural resinous mixture produced by bees. It provides beneficial effects on human health in the treatment/management of many diseases. The present study was performed to demonstrate the anti- Acanthamoeba activity of ethanolic extracts of Propolis samples from Iran. The interactions of the compounds and essential proteins of Acanthamoeba were also visualized through docking simulation. Methods: The minimal inhibitory concentrations (MICs) of Propolis extract against Acanthamoeba trophozoites and cysts was determined in vitro. In addition, two-fold dilutions of each of agents were tested for encystment, excystment and adhesion inhibitions. Three major compounds of Propolis extract such as chrysin, tectochrysin and pinocembrin have been selected in molecular docking approach to predict the compounds that might be responsible for encystment, excystment and adhesion inhibitions of A. castellanii. Furthermore, to confirm the docking results, molecular dynamics (MD) simulations were also carried out for the most promising two ligand-pocket complexes from docking studies. Results : The minimal inhibitory concentrations (MICs) 62.5 and 125 µg/mL of the most active Propolis extract were assessed in trophozoites stage of Acanthamoeba castellanii ATCC30010 and ATCC50739, respectively. At concentrations lower than their MICs values (1/16 MIC), Propolis extract revealed inhibition of encystation. However, at 1/2 MIC, it showed a potential inhibition of excystation and anti-adhesion. The molecular docking and dynamic simulation revealed the potential capability of Pinocembrin to form hydrogen bonds with A. castellanii Sir2 family protein (AcSir2), an encystation protein of high relevance for this process in Acanthamoeba. Conclusions : The results provided a candidate for the development of therapeutic drugs against Acanthamoeba infection. In vivo experiments and clinical trials are necessary to support this claim.

Background : Propolis is a natural resinous mixture produced by bees. It provides beneficial effects on human health in the treatment/management of many diseases. The present study was performed to demonstrate the anti- Acanthamoeba activity of ethanolic extracts of Propolis samples from Iran. The interactions of the compounds and essential proteins of Acanthamoeba were also visualized through docking simulation. Methods: The minimal inhibitory concentrations (MICs) of Propolis extract against Acanthamoeba trophozoites and cysts was determined in vitro. In addition, two-fold dilutions of each of agents were tested for encystment, excystment and adhesion inhibitions. Three major compounds of Propolis extract such as chrysin, tectochrysin and pinocembrin have been selected in molecular docking approach to predict the compounds that might be responsible for encystment, excystment and adhesion inhibitions of A. castellanii. Furthermore, to confirm the docking results, molecular dynamics (MD) simulations were also carried out for the most promising two ligand-pocket complexes from docking studies. Results : The minimal inhibitory concentrations (MICs) 62.5 and 125 µg/mL of the most active Propolis extract were assessed in trophozoites stage of Acanthamoeba castellanii ATCC30010 and ATCC50739, respectively. At concentrations lower than their MICs values (1/16 MIC), Propolis extract revealed inhibition of encystation. However, at 1/2 MIC, it showed a potential inhibition of excystation and anti-adhesion. The molecular docking and dynamic simulation revealed the potential capability of Pinocembrin to form hydrogen bonds with A. castellanii Sir2 family protein (AcSir2), an encystation protein of high relevance for this process in Acanthamoeba. Conclusions : The results provided a candidate for the development of therapeutic drugs against Acanthamoeba infection. In vivo experiments and clinical trials are necessary to support this claim.

The aims of this study to assess the efficiency of AGL against acetaminophen (APAP)-induced hepatic toxicity that was generated by mitochondrial oxidative stress and glutathione depletion. Free radical scavenging potentiality was analyzed by using 2, 2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide, nitric oxide, and hydroxyl radical scavenging assays. APAP-induced liver toxicity was formed at a dose level of 640 mg/kg mg/kg BW each, p.o. for 14 days for all experimental rats except the vehicle control group. AGL (5 and 10 mg/kg) were treated orally with negative control and negative control silymarin (50 mg/kg) group. To assess the protective effect, we looked at the levels of serum biochemical markers, liver histoarchitecture, and hepatic antioxidant enzyme activity. AGL showed in vitro anti-oxidant potentialities by scavenging radicals in the respective assays. As evidenced by serum biochemical indicators and relative liver weight, AGL co-administration substantially reduced toxicant-induced hepatic damage. APAP-intoxication increased the malondialdehyde (MDA) level and declined in cellular endogenous antioxidant enzymes such as reduced catalase, superoxide dismutase, and glutathione, where, AGL treatment amended their level. In the same way, histopathological evaluation further verified that AGL protected the hepatocyte from APAP-induced damage. As AGL scavenges toxic free radicals, thereby protects mitochondria and other organelles from reactive oxygen and nitrogen species-mediated stress and its eventual consequence necrosis. Therefore, we propose the hepatoprotective activity of AGL through its antioxidant mechanism.

Background: Propolis is a natural resinous mixture produced by bees. It provides beneficial effects on human health in the treatment/management of many diseases. The present study was performed to demonstrate the anti-Acanthamoeba activity of ethanolic extracts of Propolis samples from Iran. The interactions of the compounds and essential proteins of Acanthamoeba were also visualized through docking simulation. Methods: The minimal inhibitory concentrations (MICs) of Propolis extract against Acanthamoeba trophozoites and cysts was determined in vitro. In addition, two-fold dilutions of each of agents were tested for encystment, excystment and adhesion inhibitions. Three major compounds of Propolis extract such as chrysin, tectochrysin and pinocembrin have been selected in molecular docking approach to predict the compounds that might be responsible for encystment, excystment and adhesion inhibitions of A. castellanii. Furthermore, to confirm the docking results, molecular dynamics (MD) simulations were also carried out for the most promising two ligand-pocket complexes from docking studies. Results: The minimal inhibitory concentrations (MICs) 62.5 and 125 µg/mL of the most active Propolis extract were assessed in trophozoites stage of Acanthamoeba castellanii ATCC30010 and ATCC50739, respectively. At concentrations lower than their MICs values (1/16 MIC), Propolis extract revealed inhibition of encystation. However, at 1/2 MIC, it showed a potential inhibition of excystation and anti-adhesion. The molecular docking and dynamic simulation revealed the potential capability of Pinocembrin to form hydrogen bonds with A. castellanii Sir2 family protein (AcSir2), an encystation protein of high relevance for this process in Acanthamoeba. Conclusions: The results provided a candidate for the development of therapeutic drugs against Acanthamoeba infection. In vivo experiments and clinical trials are necessary to support this claim.

Background: Propolis is a natural resinous mixture produced by bees. It provides beneficial effects on human health in the treatment/management of many diseases. The present study was performed to demonstrate the anti-Acanthamoeba activity of ethanolic extracts of Propolis samples from Iran. The interactions of the compounds and essential proteins of Acanthamoeba were also visualized through docking simulation. Methods: The minimal inhibitory concentrations (MICs) of Propolis extract against Acanthamoeba trophozoites and cysts was determined in vitro. In addition, two-fold dilutions of each of agents were tested for encystment, excystment and adhesion inhibitions. Three major compounds of Propolis extract such as chrysin, tectochrysin and pinocembrin have been selected in molecular docking approach to predict the compounds that might be responsible for encystment, excystment and adhesion inhibitions of A. castellanii. Furthermore, to confirm the docking results, molecular dynamics (MD) simulations were also carried out for the most promising two ligand-pocket complexes from docking studies. Results: The minimal inhibitory concentrations (MICs) 62.5 and 125 µg/mL of the most active Propolis extract were assessed in trophozoites stage of Acanthamoeba castellanii ATCC30010 and ATCC50739, respectively. At concentrations lower than their MICs values (1/16 MIC), Propolis extract revealed inhibition of encystation. However, at 1/2 MIC, it showed a potential inhibition of excystation and anti-adhesion. The molecular docking and dynamic simulation revealed the potential capability of Pinocembrin to form hydrogen bonds with A. castellanii Sir2 family protein (AcSir2), an encystation protein of high relevance for this process in Acanthamoeba. Conclusions: The results provided a candidate for the development of therapeutic drugs against Acanthamoeba infection. In vivo experiments and clinical trials are necessary to support this claim.

Background : Propolis is a natural resinous mixture produced by bees. It provides beneficial effects on human health in the treatment/management of many diseases. The present study was performed to demonstrate the anti- Acanthamoeba activity of ethanolic extracts of Propolis samples from Iran. The interactions of the compounds and essential proteins of Acanthamoeba were also visualized through docking simulation. Methods: The minimal inhibitory concentrations (MICs) of Propolis extract against Acanthamoeba trophozoites and cysts was determined in vitro. In addition, two-fold dilutions of each of the agents were tested for encystment, excystment and adhesion inhibitions. Three major compounds of Propolis extract such as chrysin, tectochrysin and pinocembrin have been selected in molecular docking approach to predict the compounds that might be responsible for encystment, excystment and adhesion inhibitions of A. castellanii. Furthermore, to confirm the docking results, molecular dynamics (MD) simulations were also carried out for the most promising two ligand-pocket complexes from docking studies. Results : The minimal inhibitory concentrations (MICs) 62.5 and 125 µg/mL of the most active Propolis extract were assessed in trophozoites stage of Acanthamoeba castellanii ATCC30010 and ATCC50739, respectively. At concentrations lower than their MICs values (1/16 MIC), Propolis extract revealed inhibition of encystation. However, at 1/2 MIC, it showed a potential inhibition of excystation and anti-adhesion. The molecular docking and dynamic simulation revealed the potential capability of Pinocembrin to form hydrogen bonds with A. castellanii Sir2 family protein (AcSir2), an encystation protein of high relevance for this process in Acanthamoeba. Conclusions : The results obtained provided a candidate for the development of therapeutic drugs against Acanthamoeba infection. In vivo experiments and clinical trials are necessary to support this claim.

Background and Objectives: Cutaneous leishmaniasis is one of the common diseases between humans and animals that is considered a health problem in Iran and many countries around the world. Currently, glucantime is used to treat cutaneous leishmaniasis. Due to its high side effects and resistance, the use of alternative therapies, especially plants and natural compounds, has been highly recommended by researchers. The aim of this study is to investigate the effect of the methanolic extract of dandelion root (Taraxacum officinale) on Leishmania major in vitro techniques. Methods: In this experimental study, the extract was prepared by the Soxhlet method and to evaluate the effect of methanolic extract of dandelion root on L. major parasite at concentrations of 300-1200 mg/ml. Amphotericin B and distilled water were considered as positive and negative controls, respectively. Then, 106 live parasites were added to all tubes, and all groups were kept at 25 ±1 °C. At 24, 48, and 72 hours, the number of live parasites was counted by Trypan Blue using a neobar slide and light microscope (Hemocytometer method). Then Half-maximal inhibitory concentration (IC50) value for the above extract was calculated using SigmaPlot™ 13 software. All steps of the experiment were done in triplicate and the results were considered as average. Results: The IC50 content of methanolic extract of dandelion roots after 24, 48, and 72 hours on L. major were calculated to be 1.04, 0.9, and 0.68 µg/ml. The highest growth inhibition (100%) was observed at a concentration of 1200 μg/ml after 72 hours of exposure. There was a significant difference between the IC50 extract and amphotericin B drug after 24, 48 and 72 hours (P

ABSTRACT: Background: Bacterial resistance to multiple drugs has recently emerged as a serious health problem. Concomitantly, the characterization of new substances with potential antimicrobial activity has been less frequent in the drug development industry. The overexpression of genes encoding efflux pumps that expel antimicrobial drugs from the intracellular environment, lowering these to subinhibitory concentrations, are among the resistance mechanisms predisposing microorganisms to high drug resistance. Staphylococcus aureus is a bacterium found in the normal microbiota of the skin and mucous membranes, and is an opportunistic microorganism capable of causing infections with high rates of morbidity and mortality. TetK is an efflux pump characterized by its ability to provide bacterial resistance to antibiotics from the tetracycline class. This study aimed to evaluate the inhibitory effect of ferulic acid and four of its esterified derivatives against resistant Staphylococcus aureus strains. Method: Ferulic acid derivatives were obtained by esterification and then characterized by hydrogen and carbon-13 nuclear magnetic resonance analysis. The minimum inhibitory concentrations (MIC) of ferulic acid and its esterified derivatives, ethidium bromide, and antibiotics were obtained using the microdilution test, while the efflux pump inhibition test was conducted by examining reduction in the MICs. Results: Propylferulate was seen to reduce the minimum inhibitory concentration (MIC) of both the control substance ethidium bromide and the tested antibiotic, indicating that this compound is promising for the use of efflux pump inhibition of IS-58 strains. Conclusions: This study provides strong evidence that the molecular basis for this activity is potentially due to the MsrA and TetK efflux pumps. However, further investigations are necessary to prove this hypothesis and elucidate the potentiating mechanism of the modulatory effect.

Cymbopogon winterianus, known as “citronella grass”, is an important aromatic and medicinal tropical herbaceous plant. The essential oil of C. winterianus (EOCw) is popularly used to play an important role in improving human health due to its potential as a bioactive component. The present study aimed to identify the components of the essential oil of C. winterianus and verify its leishmanicidal and trypanocidal potential, as well as the cytotoxicity in mammalian cells, in vitro. The EOCw had geraniol (42.13%), citronellal (17.31%), and citronellol (16.91%) as major constituents. The essential oil only exhibited significant cytotoxicity in mammalian fibroblasts at concentrations greater than 250 μg/mL, while regarding antipromastigote and antiepimastigote activities, they presented values considered clinically relevant, since both had LC50 < 62.5 μg/mL. It can be concluded that this is a pioneer study on the potential of the essential oil of C. winterianus and its use against the parasites T. cruzi and L. brasiliensis, and its importance is also based on this fact. Additionally, according to the results, C. winterianus was effective in presenting values of clinical relevance and low toxicity and, therefore, an indicator of popular use.