Mehdi Karamian

Leishmaniasis is a neglected tropical disease that urgently requires safer and more effective therapeutic strategies. Rhoifolin (RF), a natural flavonoid with anti-inflammatory and antiparasitic potential, has shown promising biological activities against several infectious diseases. In the present study, the antileishmanial effects of free RF and its niosomal formulation (NRF), alone and in combination with meglumine antimoniate (MA), were investigated against Leishmania major. Niosomes were prepared using Span 40, Tween 40, and cholesterol, resulting in high encapsulation efficiency (90.9%) and a sustained drug-release profile. The antileishmanial activity of RF, NRF, MA, and their combinations against promastigotes was evaluated using both MTT and trypan blue exclusion assays, while anti-amastigote activity was assessed in infected macrophages. NRF exhibited greater efficacy than free RF against both promastigote and amastigote forms of L. major. Combination treatments demonstrated additive interactions (combination index ≈ 1.0) and higher selectivity indices, indicating enhanced efficacy and safety. In addition, treatment with RF, NRF, and their combinations significantly increased LDH release and ROS production in infected macrophages compared with untreated controls. Molecular docking analysis revealed a strong binding affinity of RF toward caspase-3. Furthermore, treatment with RF and NRF significantly modulated apoptosis-related mediators, including increased expression of CASP3, CASP8, and Bax, along with reduced Bcl-2 expression. Immunological analyses also demonstrated macrophage polarization toward the M1 phenotype through upregulation of TNF-α, IFN-γ, IL-12, and iNOS/NO, accompanied by downregulation of IL-10, TGF-β, and ARG1. These immunomodulatory and apoptosis-associated effects were more pronounced in the NRF and NRF + MA groups. Overall, our findings suggest that niosomal RF represents a promising lipid-based delivery system for enhancing the antileishmanial efficacy of RF. The NRF + MA combination may provide a potential therapeutic strategy for improving treatment outcomes and reducing the limitations associated with current therapies for cutaneous leishmaniasis.

Background: Snails of the Lymnaeidae family are the intermediate hosts of Fasciola species, the causative agents of fascioliasis. The purpose of this study was to determine the prevalence of Fasciola species in lymnaeid snails and to investigate the association of geoclimatic factors and Fasciola species distribution in northwestern provinces of Iran using geographical information system (GIS) data. Methods: A total of 2000 lymnaeid snails were collected from 33 permanent and seasonal habitats in northwestern Iran during the period from June to November 2021. After identification by standard morphological keys, they were subjected to shedding and crushing methods. Different stages of Fasciola obtained from these snails were subjected to the ITS1 polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) method for species identification. The associations of weather temperature, rainfall, humidity, evaporation, air pressure, wind speed, elevation, and land cover with the distribution of Fasciola species were investigated. Geographical and statistical analysis was performed using ArcMap and SPSS software, respectively, to determine factors related to Fasciola species distribution. Results: Of the 2000 snails collected, 19 were infected with Fasciola hepatica (0.09%), six with F. gigantica (0.03%), and 13 with other trematodes. Among geoclimatic and environmental factors, mean humidity, maximum humidity, and wind speed were significantly higher in areas where F. hepatica was more common than F. gigantica. The altitude of F. hepatica-prevalent areas was generally lower than F. gigantica areas. No significant relationship was observed between other investigated geoclimatic factors and the distribution of infected snails. Conclusions: The present study showed the relationship of humidity and wind speed with the distribution of snails infected with F. hepatica or F. gigantica in the northwestern regions of Iran. In contrast to F. gigantica, F. hepatica was more prevalent in low-altitude areas. Further research is recommended to elucidate the relationship between geoclimatic factors and the presence of intermediate hosts of the two Fasciola species. Graphical

Cutaneous leishmaniasis is a major health problem in several parts of the world. However, existing drugs have various therapeutic effects on different Leishmania species and also cause many side effects. This study focuses on the use of the aqueous extract of quercetin in producing silver nanoparticles for the experimental treatment of cutaneous leishmaniasis in laboratory mice. Silver nanoparticles (AgNPs) were synthesized using quercetin and characterized by UV-Vis spectroscopy, DLS, FTIR, and FESEM. We investigated the antileishmanial activity of green synthesized AgNPs in vitro and in vivo. Synthesized AgNPs from quercetin macromolecules showed promising antileishmanial activity at an IC50 of 125 μg/ml against promastigotes in vitro. In the in vivo study, L. major-infected BALB/c mice were treated topically with luteolin/quercetin-capped silver nanoparticle (AgNPQct) ointment for 21 consecutive days. As a result, this ointment reduced the inflammatory response compared to the untreated group ( < 0.05). The size of lesions in the luteolin/AgNPQct group showed a slight increase compared to other groups ( < 0.01). Biogenic silver nanoparticles along with the luteolin macromolecule can be an effective and reliable candidate for the design of antileishmanial drugs in the future.

Background: In recent years, cases of human visceral leishmaniasis (HVL) have been reported in some districts of Golestan Province, northeastern Iran, particularly in rural areas. Recent epidemiological evidence in Leishmania infantum endemic regions of in Iran indicates approximately 50%-80% of seropositive dogs are asymptomatic for Leishmania infection. Objectives: The goal in this study was to determine Leishmania species infecting domestic dogs in Golestan Province, Iran. Methods Between 2015 and 2016, blood samples were obtained from 100 domestic dogs in rural regions of Golestan Province, northeastern Iran. All samples were tested for anti-Leishmania antibodies using a direct agglutination test (DAT), and for Leishmania spp. kinetoplast DNA (kDNA) using PCR. Results: Seven (7%) dogs were antibody positive and 25 dogs (25%) were Leishmania spp. DNA positives by PCR positive for leishmaniasis. Four of the seven (71%) antibody-positive dogs and 19 of the 25 (76%) PCR-positive dogs were asymptomatic. The rate of infection detected by PCR was significantly higher in male dogs (21/75, 28%) than that in female dogs (4/25, 16%). The ITS1 PCR-RFLP assay identified the presence of L. infantum, L. tropica or Crithidia spp. in the 25 PCR-positive samples. Conclusions: The high proportion of asymptomatic dogs in the study areas represent they act as potential reservoirs in the transmission cycle of Leishmania spp. and also Crithidia fasciculata as an emerging agent for the first time. Moreover, our data showed that PCR is a more reliable assay than DAT for detecting Leishmania spp. infection among asymptomatic dogs.

Background: Cutaneous leishmaniasis (CL) has been reported in recent years in South Khorasan Province, a desert region of eastern Iran, where the main species isLeishmania tropica. Little is known of the influence of geography and climate on its distribution, and so this study was conducted to determine geo-climatic factors by using geographic information system.Methods: The home addresses of patients with CL patients who were diagnosed and notified from 2009 to 2017 were retrieved from the provincial health center and registered on the village/town/city point layer. The effects of mean annual rainfall (MAR) and mean annual humidity (MAH), mean annual temperature (MAT), maximum annual temperature (MaxMAT), minimum annual temperature (MinMAT), mean annual number of high-velocity wind days (MAWD), mean annual frosty days (MAFD) and snowy days (MASD), elevation, soil type and land cover on CL distribution were examined. The geographical analysis was done using ArcMap software, and univariate and multivariate binary logistic regression were applied to determine the factors associated with CL.Results: A total of 332 CL patients were identified: 197 (59.3%) male and 135 (40.7%) female. Their mean age was 29.3 ± 2.1 years, with age ranging from 10 months to 98 years. CL patients came from a total of 86 villages/towns/cities. By multivariate analysis, the independent factors associated with increased CL were urban setting (OR = 52.102), agricultural land cover (OR = 3.048), and MAWD (OR = 1.004). Elevation was a protective factor only in the univariate analysis (OR = 0.999). Soil type, MAH, MAT, MinMAT, MaxMAT, and MAFD did not influence CL distribution in eastern Iran.Conclusions: The major risk zones for CL in eastern Iran were urban and agricultural areas with a higher number of windy days at lower altitudes. Control strategies to reduce human vector contact should be focused in these settings.Graphical

Cutaneous leishmaniasis (CL) is one of the most important health challenges in hyperendemic countries like Iran. Geospatial information systems‐based studies have shown that factors, including land cover, altitude, slope temperature, rainfall and animal livestock, affect CL distribution in Kohgyloyeh and Boyerahmad province, southwestern Iran. However, the question of the influence of nomadic tribes, who travel with their goats and sheep, on CL is unanswered. We, therefore, investigated their role in CL epidemiology from 2008 to 2017 and compare them with geoclimatic factors. CL patient demographic data and their village/city addresses were retrieved from Provincial Health Center and mapped on the geographic information system (GIS) layer of the province's political divisions. Nomadic travel routes (NTRs) with a 2 km buffer were generated and their effect on CL was investigated together with the interpolated layers of rainfall, temperatures, humidity, slope, elevation, land covers, by binary regression. CL was significantly more common in villages/cities in the 2 km NTR zone (p value < .001; OR = 1.96; 95% CI = 1.4–2.745). Geoclimatic factors, including slope, elevation, rainfall, temperatures, humidity and most of the landcovers, were not significantly different inside and outside the NTR. Areas of irrigated farm were the only effective landcover on CL (p value = .049; OR = 2.717; 95% CI = 1.003–7.361) within the NTR versus non‐NTR. Living within NTRs almost doubled the risk of acquiring CL. Several factors for this include passage through areas of high sand fly activity, increased contact between sandflies and humans, sheep and goats, and feeding on their blood and faeces, and low availability of health facilities that should be more investigated and considered in the future control programs.

Cutaneous leishmaniasis caused by Leishmania major (L. major) is an endemic disease in Iran. The current reference drugs, including Glucantime, possess high toxicity in addition to some side-effects. Therefore, there is a growing interest in exploring biomedical plants. The goal of the present study was to evaluate the anti-leishmanial activity and cytotoxicity of hydroalcoholic extracts from Prosopis farcta (P. farcta) over promastigote and amastigote forms. This study was performed at the Iran Birjand University of Medical Sciences, during the year 2019. In this study, the hydroalcoholic extracts of the stems, leaves (LE) and fruits (FE) of P. farcta were obtained. The anti-leishmanial activity was assessed against leptomonad promastigotes and intracellular amastigotes of L. major. The cytotoxicity of these extracts was determined in murine macrophages. The FE and LE of P. farcta demonstrated a significant leishmanicidal effect against L. major promastigotes with an IC50 of 0.9 mg/mL and 1.1 mg/mL, respectively. The FE showed the most anti-leishmanial activity and presented with the highest index of selectivity (SI=14.6) as an anti-leishmanial product. Infected macrophages treated using the FE showed a reduction in parasite burden by 97.3%. The results of the present study demonstrated the leishmanicidal activity of P. farcta on both promastigotes and intracellular amastigotes. There is a need for performing comprehensive studies on relevant animal models and to access the effects of active components of P. farcta extract on the growth of L. major.

Background and Objectives: Fascioliasis is a common disease of human and animals caused by Fasciola hepatica and F. gigantica. The life cycle of these trematodes is influenced by climatic and geographical factors. The aim of this study was to determine the geographical features of animal fascioliasis in Kohgiluyeh and Boyer-Ahmad province ...

Background: Trichostrongylus is one of the most important zoonotic trichostrongylid nematodes, infecting mostly livestock. Data on its genetic characteristics are lacking in Iran. Methods: We determined the phylogenetic relationships of Trichostrongylus species in three counties of Kohgiloyeh and Boyerahmad (K-B) province, southwest Iran. Small intestine and abomasum of 70 sheep and goats were investigated. Results: A total of 35 isolates of Trichostrongylus worms were detected and all were genetically identified as Trichostrongylus vitrinus. Analysis of 321 bp of the internal transcribed spacer 2 (ITS2) of ribosomal DNA revealed 16 genotypes. All genotypes were single nucleotide polymorphisms, including some hypervariable points. All sequences were trimmed to 170 bp, compared with sequences on GenBank including short sequences from other endemic foci of Iran and other countries and all isolates were used to generate a maximum likelihood phylogenetic tree, which consisted of two clades A and B. Clade A included isolates from Iran, Russia, New Zealand, Australia and the UK; clade B only contained South African isolates. Most clade A isolates (north, southwest and west Iran, Russia, New Zealand, Australia and UK) were in a similar phylogenetic position. One subclade was detected in clade A (isolates from Southwest Iran, New Zealand and UK). Conclusions: We hypothesize that drug resistant T. vitrinus may account for its exclusive detection in our samples. The high similarity of genotypes from Iran, New Zealand and UK may be due to their close political relationships during the colonial era. More research is needed to understand better the phylogeny of T. vitrinus and its relationship with drug resistance and human transmission.