Background Leishmaniasis is a neglected infectious disease caused by protozoa of the genus Leishmania. The disease generally manifests as characteristic skin lesions which require lengthy treatment with antimonial drugs that are often associated with adverse side effects. Therefore, a number of studies have focused on natural compounds as promising drugs for its treatment. This study aimed to evaluate the effects of larval excretion/secretion products (ES) of Lucilia sericata in crude and fractionated forms on Leishmania major, by using in vitro and in vivo models. Methods The in vitro experiments involved evaluation of ES on both promastigotes and macrophage-engulfed amastigotes, whereas the in vivo experiments included comparative treatments of skin lesions in L. major-infected mice with Eucerin-formulated ES and Glucantime. Results The half maximal inhibitory concentrations of the crude ES, > 10-kDa ES fraction, < 10-kDa ES fraction, and Glucantime were 38.7 μg/ml, 47.6 μg/ml, 63.3 μg/ml, and 29.1 μg/ml, respectively. Significant differences were observed between percentage viabilities of promastigotes treated with the crude ES and its fractions compared with the negative control (P < 0.0001). The crude ES was more effective on amastigotes than the two ES fractions at 300 μg/ml. The macroscopic measurements revealed that the reduction of lesion size in mice treated with the crude ES followed quicker cascades of healing than that of mice treated with Glucantime and the ES fractions. Conclusions The present study showed that the larval ES of L. sericata in both crude and fractionated forms are effective for both intracellular and extracellular forms of L. major. Also, the ES exert both topical and systemic effects on mice experimentally infected with L. major. Graphical abstract

Background: Microsporidia are spore-forming intracellular pathogens with worldwide prevalence, causing emerging infections in humans and animals. Enterocytozoon bieneusi is a zoonotic species of microsporidia and is responsible for more than 90% of cases of microsporidiosis in humans and animals. Pigs and wild boars are important animal reservoirs of microsporidia. Hence, we aimed to estimate the global prevalence of microsporidia and genetic diversity of E. bieneusi in pigs and wild boars through a set of systematic review and meta-analysis (PRISMA) guidelines.Methods: Four databases (Web of Science, PubMed, Scopus and Google Scholar) were searched between January 1, 2000 and April 30, 2021. Regarding meta-analysis, the random-effect model was employed by forest plot with 95% confidence interval (CI).Results: After exclusion of irrelevant articles and duplication removal, 33 papers, including 34 datasets (30 datasets for domestic pigs and 4 for wild boars) finally meet the inclusion criteria to undergo meta-analysis. The pooled prevalence rates of microsporidia infection in domestic pigs and wild boars were 37.6% (95% CI: 30.8-44.9%) and 8.1% (95% CI: 2.1-26.8%), respectively. While, the pooled prevalence rates of E. bieneusi were 35% (95% CI: 28.4-42.2%) in domestic pigs and 10.1% (95% CI: 1.7-42.4%) in wild boars. The genotypes EbpA was the most reported genotype in domestic pigs and wild boars. Male animals had higher prevalence rates of microsporidia infection than females (27 vs. 17.4%, OR = 1.91; 95% CI, 0.77-4.71%). Conclusion:This study indicates the important role of domestic pigs and wild boars as animal reservoir hosts of microsporidia. Thereby, strategies for control and prevention of these zoonotic pathogens should be designed in pigs and wild boars.

Leishmaniasis is a zoonotic disease caused by an intracellular parasite from the genus Leishmania. Lack of safe and effective drugs has increasingly promoted researches into new drugs of natural origin to cure the disease. The study, therefore, aimed to investigate the anti-leishmanial effects of Lucilia sericata larval excretion/secretion (ES) in combination with Apis mellifera honey as a synergist on Leishmania major using an in vitro model. Various concentrations of honey and larval ES fractions were tested against promastigotes and intracellular amastigotes of L. major using macrophage J774A.1 cell line. The inhibitory effects and cytotoxicity of ES plus honey were evaluated using direct counting method and MTT assay. To assess the effects of larval ES plus honey on the amastigote form, the rate of macrophage infection and the number of amastigotes per infected macrophage cell were estimated. The 50% inhibitory concentration (IC50) values were 21.66 μg/ml, 43.25 60 μg/ml, 52.58 μg/ml, and 70.38 μg/ml for crude ES plus honey, ES >10 kDa plus honey, ES <10 kDa plus honey, and honey alone, respectively. The IC50 for positive control (glucantime) was 27.03 μg/ml. There was a significant difference between viability percentages of promastigotes exposed to different doses of applied treatments compared to the negative control (p≤ 0.0001). Microscopic examination of amastigote forms revealed that dosages applied at 150 to 300 μg/ml significantly reduced the rate of macrophage infection and the number of amastigotes per infected macrophage cell. Different doses of larval products plus honey did not show a significant toxic effect agaist macrophage J774 cells. The larval ES fractions of L. sericata in combination with A. mellifera honey acted synergistically against L. major.

Background Leishmaniasis is a neglected infectious disease caused by a kinetoplastid protozoan. The disease generally manifests as characteristic skin lesions. Due to the lack of definitive treatment and drugs without side effects, many studies have focused on natural compounds as promising drugs for its treatment. This study aimed to evaluate the effects of larval excretion/secretion products (ES) of Lucilia sericata in crude and fractionated forms on Leishmania parasites under both in vitro and in vivo conditions. Methods In vitro experiments involved evaluation of ES products on both promastigotes and amastigotes inside infected macrophages, whereas in vivo experiments included comparative treatments of Leishmanial lesions of mice using Eucerin-formulated ES products and glucantime. Results The IC50 values were 38.7 µg/ml, 47.6 µg/ml, 63.3 µg/ml, and 29.1 µg/ml for crude ES, over 10 kDa ES-fraction, under 10 kDa ES-fraction, and glucantime respectively. Significant differences were observed between viability percentages of promastigotes treated with crude ES and its fractions compared to negative control (p < 0.0001). Crude ES was more effective on amastigote than other two ES fractions at 300 µg/ml concentration. Macroscopic measurement of lesion sizes revealed that the reduction of lesion size in mice treated with crude ES followed quicker cascades of healing than in those treated with glucantime and fractionated ES. Conclusion The present study showed that larval ES of Lucilia sericata in both crude and fractionated forms are effective on both intracellular and extracellular forms of L. major. It also provided evidence that the larval ES exerts both topical and systemic therapeutic effects on leishmanial lesions of the model animal.

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