Eugenol Essential Oil and Nanoemulsion as Antihydatic Agents with Antifibrotic and Immunomodulatory Effects in Cystic Echinococcosis.

Conventional scolicidal agents are still unsatisfactory in combating hydatid disease due to their low efficacy and increased drug side effects. Therefore, novel scolicides are required. This study aimed to evaluate the antihydatic and immunomodulatory effects of eugenol essential oil (Eug) and its nanoemulsion (Eug-NE) in cystic echinococcosis (CE). Eug and Eug-NE were administered orally to CE-infected rats and compared to albendazole (ABZ). Hydatid cyst development was assessed based on organ weight and hypertrophy indicators of the infected organs, along with a histopathological and histochemical evaluation of collagen content. The immunomodulatory effects of treatment on CE were evaluated by serum cytokine levels measurement of interferon-γ (IFN-γ) and interleukin (IL)-4 and immunohistochemical (IHC) analysis of signal transducer and activator of transcription 4 (STAT4) and GATA-binding protein 3 (GATA3) markers. Eug-NE was the most effective in reducing the cyst weights, organ weights, and hypertrophy indicators and improving histopathological lesions with reduced collagen content. Eug and Eug-NE significantly increased the IFN-γ levels and decreased the IL-4 levels, while IHC analysis demonstrated a significant reduction in STAT4 and GATA3 expression in all treated groups. Eug and Eug-NE demonstrated antihydatic and preventative effects, with a substantial decrease in liver fibrosis compared to that of ABZ. Besides their promising immunomodulatory effects, their good treatment response suggests their use as alternatives or complementary scolicidal agents in hydatid cyst treatment.

New perspectives for fascioliasis in Upper Egypt's new endemic region: Sociodemographic characteristics and phylogenetic analysis of Fasciola in humans, animals, and lymnaeid vectors.

BACKGROUND: Fascioliasis is a significant vector-borne disease that has emerged in numerous tropical and subtropical countries causing severe health problems. Egypt is one of the fascioliasis endemic regions; however, the current situation in Upper Egypt is understudied, with only sporadic human cases or outbreaks. This study aims to highlight the sociodemographic characteristics of human fascioliasis in a newly emerged endemic area in Upper Egypt, along with risk factors analysis and the molecular characteristics of the fasciolid population in humans, animals, and lymnaeid snails. METHODOLOGY/PRINCIPAL FINDINGS: The study reported Fasciola infection in patients and their close relatives by analyzing the risk of human infection. Morphological and molecular characterization was performed on lymnaeid snails. Multigene sequencing was also used to characterize fasciolids from human cases, cattle, and pooled snail samples. The study identified asymptomatic Fasciola infection among family members and identified the presence of peridomestic animals as a significant risk factor for infection. This is the first genetic evidence that Radix auricularia exists as the snail intermediate host in Egypt. CONCLUSIONS/SIGNIFICANCE: This study revealed that Assiut Governorate in Upper Egypt is a high-risk area for human fascioliasis that requires additional control measures. Fasciola hepatica was the main causative agent infecting humans and snail vectors in this newly emerged endemic area. In addition, this is the first report of R. auricularia as the snail intermediate host transmitting fascioliasis in Upper Egypt. Further research is required to clarify the widespread distribution of Fasciola in Egypt's various animal hosts. This provides insight into the mode of transmission, epidemiological criteria, and genetic diversity of fasciolid populations in Upper Egypt.\.

Amoebicidal effect of Allium cepa against Allovahlkampfia spelaea: A keratitis model.

Allovahlkampfia spelaea (A. spelaea) is a free-living amoeba, proved to cause Acanthamoeba-like keratitis with quite difficult treatment. This study aimed to evaluate the amoebicidal effect of Allium cepa (A. cepa) on A. spelaea trophozoites and cysts both in vitro and in vivo using Chinchilla rabbits as an experimental model of this type of keratitis. Chemical constituents of the aqueous extract of A. cepa were identified using Liquid Chromatography-mass Spectrometry (LC-MS). In vitro, A. cepa showed a significant inhibitory effect on trophozoites and cysts compared to the reference drug, chlorhexidine (CHX) as well as the non-treated control (P < 0.05) with statistically different effectiveness in terms of treatment durations and concentrations. No cytotoxic effect of A. cepa on corneal cell line was found even at high concentrations (32 mg/ml) using agar diffusion method. The in vivo results confirmed the efficacy of A. cepa where the extract enhanced keratitis healing with complete resolution of corneal ulcers in 80% of the infected animals by day 14 (post infection)pi compared to 70% recovery with CHX after 20 treatment days. The therapeutic effect was also approved at histological, immune-histochemical, and parasitological levels. Our findings support the potential use of A. cepa as an effective agent against A. spelaea keratitis.

Ganoderma lucidum extract inhibits Schistosoma mansoni survival in silico and in vitro study.

Schistosomosis is a worldwide tropical disease primarily caused by Schistosoma mansoni. Praziquantel is the only available drug for controlling schistosomosis, with many challenges. This study aims to evaluate the in vitro anti-Schistosoma effect of Ganoderma lucidum (G. lucidum) against adult and larval stages of Schistosoma based on the prediction of the binding activity of G. lucidum protein with proteins of various stages of S. mansoni by molecular docking to confirm its inhibitory potential through an insilico study. Results showed that Leu143, Ser165, Met214, and Asn213 were the primary crucial amino acids involved in the binding, with a promising large area of interactions between the two studied proteins. The in vitro study evaluated the motility and survival of adult and larval stages, compared to praziquantel and niclosamide, respectively. There was a significant reduction in the motility of adults after the two-hour incubation, with all concentrations and 100% death of all parasites with the minimal concentration (10 µg/ml) within 4 and 6 h of incubation (P<0.01). Regarding the cercariae, at a concentration of 10 µg/ml, all the cercariae (100%) died (P<0.01) after 15 min, and the miracidial complete mortality rate (100%) (P<0.01) occurred at a concentration of 10 µg/ml after 8 min. This study first predicted the binding activity of G. lucidum protein with proteins of S. mansoni at various stages and proved the anti-Schistosoma effect of G. lucidum in vitro, considered a promising treatment for schistosomosis.

Evaluation of the scolicidal activities of eugenol essential oil and its nanoemulsion against protoscoleces of hydatid cysts.

BACKGROUND: Cystic echinococcosis caused by the cestode Echinococcus granulosus remains a serious helminthic zoonosis affecting humans and animals in many endemic developing countries. Surgical intervention is the best management choice, although it is associated with high recurrence rates and serious complications. Also, the commonly used chemotherapeutics exhibited serious side effects. This study aimed to evaluate the protoscolicidal effects of eugenol (Eug) essential oil and its nanoemulsion (Eug-NE) against protoscoleces (PCs) of hydatid cysts in vitro. METHODS: Eug-NE was prepared and characterized. Their cytotoxicity on macrophages was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. E. granulosus PCs were treated with various concentrations of Eug and Eug-NE at different exposure times. The viability of protoscoleces was evaluated by the eosin exclusion test, and the changes in the morphology of protoscoleces were assessed. Albendazole (ABZ) was used as a positive control. RESULTS: The cellular cytotoxicity of Eug and Eug-NE on macrophage cells, in minimum and maximum concentrations (0.2 and 1 µl/mL), were nearly negligible ranging from 4.7% to 8.3% and 3.7% to 7.2%, respectively. The results showed highly significant activity of Eug-NE and Eug against hydatid PCs compared to ABZ (P < 0.05). Eug and Eug-NE have similar protoscolicidal effects at all used concentrations. Their highest scolicidal activity (100% mortality rate) was recorded at 1 µl/ml after 30 min incubation (LC50 = 0.298-LC90 = 0.521 and LC50 = 0.309-LC90 = 0.646, respectively). Both formulations showed time- and dose-dependent effects. CONCLUSIONS: This study suggested the potent scolicidal activities of Eug and Eug-NE as promising alternative scolicidal agents. Future studies are recommended to explore the mechanism of action and treatment response in vivo and clinical settings.

The protective effect of Lactobacillus acidophilus on experimental animals challenged with Trichinella spiralis; new insights on their feasibility as prophylaxis in Trichinella spiralis endemic area.

Trichinellosis is a common parasitic zoonosis. Complications of anthelmintic drugs combined with steroids raise the urge of alternative protective ways. The study aimed to investigate the protective effects of Lactobacillus acidophilus probiotic on both Trichinella spiralis adults and larvae in experimental animal models. Thirty-six male BALB/c mice were divided into 3 groups: negative control Group (G I); Group (G II) mice were inoculated orally by 500 Trichinella spiralis larvae; tested Group (G III) mice were prophylactic by an oral dose of Lactobacillus acidophilus in commercially available form for seven consecutive days, before infection. Mature worms and encysted larvae were counted on the 5th and 21st day post-infection (dpi), respectively. IL-1, IL-6, IL-10 and TNF-α concentrations were estimated at 5th and 21st dpi of all groups. Significant reductions in mean worms and larvae burden were detected by 62.1% and 73.5% in the prophylactic group compared to the non-prophylactic group. The cytokine profiles were revealed IL-1 and IL-6 up-regulation compared to IL-10 and TNF-α down-regulation in the tested group compared to other groups. Although Lactobacillus acidophilus failed to achieve complete eradication of Trichinella spiralis adults and larvae, it showed powerful effects in reducing parasites and cytokines burdens.

Thymus vulgaris, Mentha piperita and Elettaria cardamomum against Trypanosoma evansi in vitro and in an animal model with new insights for the treatment of trypanosomosis.

Trypanosomosis is a worldwide disease that affects human and livestock populations with limited availability and high cost of trypanocides. The study aims to evaluate the possible in vitro and in vivo anti-trypanosomal activity of Thymus vulgaris (thyme), Mentha piperita (mint) and Elettaria cardamomum (cardamom) aqueous extracts against Trypanosoma evansi in experimentally infected rats using Intropar as a reference drug. The crude extracts of the selected plants were used in three concentrations (2500, 2000, and 1000 µg/ml). The in vitro trypanocidal activities were assessed regarding parasite motility, count, and infectivity. The in vivo susceptibility of T. evansi was evaluated by assessing the level of parasitemia in the experimental rats. The packed cell volume (PCV) was also monitored. Both the in vitro and in vivo experiments showed trypanocidal activity, of all the tested extracts, higher than that of Intropar. The in vitro trypanocidal effects were dose-dependent and represented by a significant reduction of the parasite count together with immobilizing effect within 3 hours incubation period, compared to the negative and positive controls (p< 0.05). The in vivo trypanocidal effects of the different concentrations of all the tested extracts were represented by the significantly lowered levels of parasitemia, compared to the negative control (NC) group with varying degrees; in a dose-dependent manner concerning the time. They exhibited also a significantly higher level in PCV recovery compared to the NC group (p < 0.05). This study initially confirmed the potent in vitro and in vivo trypanocidal effect of the three extracts, with a potentially promising future for the treatment of trypanosomosis.

Green biosynthesis of silver nanoparticles by Aspergillus niger and its antiamoebic effect against Allovahlkampfia spelaea trophozoite and cyst.

BACKGROUND: Fungi represent an interesting candidate for the synthesis of nanoparticles. The biosynthesis of silver nanoparticles (AgNPs) has many industrial and biomedical indications. We aimed in this work to biologically synthesize silver nanoparticles using Aspergillus niger and to evaluate its effect against the newly identified Allovahlkampfia spelaea that causes resistant human keratitis. MATERIAL AND METHODS: Aspergillus niger (soil isolate) was treated with silver nitrate to produce silver nanoparticles. AgNPs were characterized by Ultraviolet-Visible Spectroscopy, Transmission Electron Microscopy, and Fourier Transform Infrared Spectroscopy. The effect of the synthesized nanoparticles against Allovahlkampfia spelaea growth, encystation, excystation, and toxicity in host cells was evaluated. RESULTS: AgNPs exhibited significant inhibition of Allovahlkampfia spelaea viability and growth of both trophozoites and cysts, with a reduction of amoebic cytotoxic activity in host cells. CONCLUSION: AgNPs may give a promising future to the treatment of Allovahlkampfia spelaea infections in humans.

Morphological and initial molecular characterization of Clogmia albipunctatus larvae (Diptera: Psychodidae) causing urinary myiasis in Egypt.

Myiasis is the infestation of human tissues by dipterous fly larvae of the class Insecta. Clogmia albipunctatus, family Psychodidae, is one of the most medically important insects that cause human myiasis. The aim of the present study is the morphological identification and the molecular characterization of moth flies causing many cases of urinary myiasis in Egypt, based on sequencing of the mitochondrial DNA of the larvae. Seven urinary samples of patients complaining of urinary symptoms and giving a history of low socioeconomic level were examined. Recovered larvae were identified using light microscopy and SEM. For molecular identification, the mitochondrial genes Cytochrome B (cytB), NADH1, NADH1, and 16S were sequenced and phylogenetically analyzed. The morphological and molecular characterization could accurately diagnose our patients to have C. albipunctatus infestation. Such results provided the initial set of data on the molecular identification and phylogenetic analysis of moth flies based on DNA barcoding in Egypt.

Allovahlkampfia spelaea Causing Keratitis in Humans.

BACKGROUND: Free-living amoebae are present worldwide. They can survive in different environment causing human diseases in some instances. Acanthamoeba sp. is known for causing sight-threatening keratitis in humans. Free-living amoeba keratitis is more common in developing countries. Amoebae of family Vahlkampfiidae are rarely reported to cause such affections. A new genus, Allovahlkampfia spelaea was recently identified from caves with no data about pathogenicity in humans. We tried to identify the causative free-living amoeba in a case of keratitis in an Egyptian patient using morphological and molecular techniques. METHODS: Pathogenic amoebae were culture using monoxenic culture system. Identification through morphological features and 18S ribosomal RNA subunit DNA amplification and sequencing was done. Pathogenicity to laboratory rabbits and ability to produce keratitis were assessed experimentally. RESULTS: Allovahlkampfia spelaea was identified as a cause of human keratitis. Whole sequence of 18S ribosomal subunit DNA was sequenced and assembled. The Egyptian strain was closely related to SK1 strain isolated in Slovenia. The ability to induce keratitis was confirmed using animal model. CONCLUSIONS: This the first time to report Allovahlkampfia spelaea as a human pathogen. Combining both molecular and morphological identification is critical to correctly diagnose amoebae causing keratitis in humans. Use of different pairs of primers and sequencing amplified DNA is needed to prevent misdiagnosis.

Resistance and Susceptibility to Malarial Infection: A Host Defense Strategy against Malaria.

BACKGROUND: In an effort to understand what limits the virulence of malaria parasites in relation to the host genetic and immunogenic background, we investigated the possibility that the parasite and host genotype crossover interactions constrain virulence. METHODS: Two groups of mice from different genotypes were used (C57BL/6 (B6) and DBA/2 mice). The mice were infected with a virulent parasite line Plasmodium yoelii 17XL (P. yoelii 17XL). Parasitemia, hematocrit value and lymphocytes yielded by livers and spleens were evaluated. Fluorescence Activated Cell Sorting (FACS) analysis illustrated phenotypic characterization of lymphocytes. RESULTS: Infection with P. yoelii 17XL did not result in the death of DBA/2 mice. In contrast, B6 mice developed significantly high parasitemia and succumbed to death. Using (FACS) analysis, DBA/2 mice were found to experience a marked expansion of interleukin (IL)-2Rß(+) CD3(int) cells and γδ T cells in the liver, especially in the recovery phase. The expansion of unconventional T cells (i.e. B220(+) T cells) was also marked in DBA/2 mice. CONCLUSION: The outcome of murine malaria infections depends on the dynamic interplay between the immune-mediator and the genotype of the host.