mehdi mohebali

Rodents are the primary reservoir hosts for zoonotic cutaneous leishmaniasis (ZCL) caused by Leishmania major. Knowing reservoir hosts is crucial for leishmaniasis surveillance and control programs in endemic areas. In this study, we examined an archived spleen of Rattus norvegicus obtained during a pest control program in 2000 in Tehran, the capital of Iran. The sample was analyzed using polymerase chain reaction (PCR) and sequencing to determine the presence of Trypanosomatidae based on the internal transcribed spacer (ITS) 1 gene. Amplification and sequencing of the discriminative region of the ITS1 gene followed by BLAST analysis showed the highest similarity with L. major isolates. Also, the phylogenetic analysis revealed that our sample was grouped with L. major isolates retrieved from the GenBank database. This finding might support the claim that R. norvegicus acts as a potential reservoir host for L. major. Further studies, including a survey on more rodent samples as well as studying sandflies in the area, might uncover the possible presence of such pathobiological conditions in ZCL transmission in urban and suburban settings.

Strongyloides stercoralis is one of the soil-transmitted helminths (STH) in tropical and subtropical regions. The role of lipid profile has been investigated in the survival of larval stages of helminths, but there is limited information about the role of lipid profiles and strongyloidiasis. Hence, we aimed to investigate the seroprevalence of S. stercoralis infection in patients with hyperlipidemia is compared with the nonhyperlipidemia.

In 2023, participants were selected from the laboratory of Porsina Hospital in Guilan Province, northern Iran and their lipid profiles including TG, CHOL, LDL, HDL, and VLDL were measured. They were divided into two groups of case and control and matched based on sex and age. S. stercoralis Ab (IgG) was measured by ELISA methods, using the NovaTec kit. Finally, statistical analysis was performed.

Each case and control group consisted of 105 participants, from 13 to 80 years old. 56.66% were female and 43.33% were male. The sero-prevalence of S. stercoralis was found 4.76% in the case group compared to 0.95% in the control group. We found an association between TG fall and VLDL with sero-prevalence of S. stercoralis in hyperlipidemia group (P= 0.034), but other lipid profiles did not show a significant association. A significant relationship was found between contact with dogs and seroprevalence of S. stercoralis (P=0.001).

The sero-prevalence of S. stercoralis in the case group was 5 times higher than the control group. A significant association between TG and VLDL fall with S. stercoralis Ab (IgG) was observed, but future studies with more sample sizes are suggested to investigate the anti-atherogenic effect of S. stercoralis. Also, a genetic assessment of S. stercoralis and the host (humane and dogs) is recommended to research zoonotic potential in epidemic areas.

Due to the increasing cohabitation of humans with infected animals, such as pets or wildlife, human infection with helminthic parasites has become an important emerging health concern. This study aimed to investigate the prevalence of helminthic parasite infections in canines in the Mughan Plain area of Ardabil Province, Iran. A total of 87 samples were collected from different canines, including stray dogs, foxes, and jackals, in the Mughan Plain area of Ardabil Province, Iran. The organs of the animals were sectioned and examined macroscopically for helminth parasitic infections. Isolated helminth species were morphologically identified using a valid identification key. The results showed that that Mesocestoides sp. was the most prevalent helminth species, with a frequency of 30 in canids (34.5%), including 8 red foxes (Vulpes vulpes, 53.33%), 12 jackals (Canis aureus, 48%), and 10 stray dogs (Canis familiaris, 21.3%). In addition, a high frequency of Toxascaris leonina was observed in 13 canines (14.9%), including 2 foxes (13%), 4 jackals (16%), and 7 wild dogs (14.9%). The prevalence of Echinococcus granulosus was reported in 9 canines (10.3%), including 1 jackal (4%) and 8 wild dogs (17.02%). The results of this study indicate that Mesocestoides sp. is the most common helminth species in canids. Our findings indicate that canines in Iran serve as both intermediate and final definitive hosts for several harmful parasites, posing a risk to humans and animals health. Further research and interventions are needed to better understand the transmission dynamics and to develop effective strategies for helminth control in canids.

Current medications especially the pentavalent antimonial compounds have been used as the first line treatment of cutaneous leishmaniasis (CL), but they have limitations due to serious side effects such as drug resistance, cardio and nephrotoxicity, and high costs. Hence, the demand to find more usable drugs is evident. Synthesis and devel-opment of natural, effective, biocompatible, and harmless compounds against Leishmania major is the principal priority of this study.

By electrospinning method, a new type of nanofiber were synthesized from royal jelly and propolis with dif-ferent ratios. Nanofibers were characterized by Scanning Electron Microscope (SEM), Transmission Electron Micros-copy (TEM), Thermogravimetric Analysis (TGA), Contact angle, and Fourier-transform infrared spectroscopy (FTIR). The Half-maximal inhibitory concentration (IC50), Half-maximal effective concentration (EC50) and the 50% cytotoxic concentration (CC50) for different concentrations of nanofibers were determined using quantitative calorimetric meth-ods. Inductively coupled plasma-optical emission spectrometry (ICP-OES) and flow cytometry were performed as com-plementary tests.

The results showed that the proposed formulas provide a new achievement that, despite the significant killing activity on L. major, has negligible cytotoxicity on the host cells. Royal jelly nanofibers have significantly shown the best 72 hours results (IC50= 35 μg/ml and EC50=16.4 μg/ml) and the least cytotoxicity.

This study presents a great challenge to introduce a new low-cost treatment method for CL, accelerate wound healing, and reduce scarring with minimal side effects and biocompatible materials. Royal jelly and propolis nanofibers significantly inhibit the growth of L. major in-vitro.

Phlebotomus papatasi (Diptera: Psychodidae) is the main vector of zoonotic cutaneous leishmaniasis (ZCL) in Iran. The nonstandard use of pesticides against pests, particularly in agriculture, indirectly has caused the de-velopment of resistance and, consequently, the threat of control measures in ZCL endemic areas. Up to 2023, several reports of resistance in Ph. papatasi have been declared in the Old World. The purpose of this study was to measure the lethal time (LT50 and LT90) of Ph. papatasi sand flies in the ZCL endemic center of Esfahan to DDT and deltamethrin insecticides.

Sand flies were collected in Borkhar and were tested using WHO adult mosquito test kit against DDT 4% and deltamethrin 0.0002%. The sand fly’s survival was recorded during exposure time in 225, 450|, 900, 1800, and 3600-seconds’ intervals for DDT and Deltamethrin and they were allowed to recover for 24 hours. Then LT50 and LT90 were analyzed using probit software. Phlebotomus papatasi were identified using morphological keys and other sand flies’ species were excluded from the analysis.

The insecticide against female Ph. papatasi revealed hundred percent mortality when exposed to DDT 4% and deltamethrin 0.0002%. The LT50 and LT90 were 19.32 and 22.74 minutes for DDT 4% and 39.92 and 51.33 minutes for deltamethrin 0.0002% respectively.

Results of this study revealed that Ph. papatasi is still susceptible to DDT and deltamethrin. This data pro-vides valuable knowledge to implement effective control strategies against ZCL main vector and help to manage insec-ticide resistance in the region.

Intestinal parasitic infections are still a considerable global public health problem. We aimed to determine the frequency of intestinal parasitic infections among people referring to the central laboratory of Meshkin Shahr City, Ardabil Province, Iran.

In this cross-sectional survey, 460 fecal samples were collected randomly from persons referred to the central laboratory of Meshkin Shahr City, from January to June 2022. The samples were examined by direct wetmount, Trichrome and modified Ziehl-Neelsen staining, formalin ethyl acetate sedimentation, and agar plate culture.

The frequency of intestinal parasites was 15.7% (72 out of 460 cases), with some people with numerous intestinal parasites. The frequency of protozoan infections (13.9%) was higher than the helminthic infections (2.6%). Blastocystis spp. (8.1%) was the most prevalent detected intestinal protozoan. Entamoeba coli (5.7%), Dicrocoelium dendriticum (2.2%), Giardia lamblia (1.5%), Fasciola spp. (0.2%), and Hymenolepis nana (0.2%) were other detected parasites.

In- spite of betterment of the health condition in Iran and reduction of parasitic infection, intestinal parasitic infections are still a considerable public health issue in some parts of Iran.

Cutaneous leishmaniasis (CL) is a parasitic disease that presents a broad spectrum of clinical features. Treatment of CL is problematic. We aimed to compare the field therapeutic efficacy of topical nanoliposomes containing 0.4% amphotericin B (Nano Lip-AmB) alone and in combination with cryotherapy and/or Glucantime® on human CL in the endemic areas of Iran.

This retrospective study was performed based on the results of using Nano Lip-AmB alone or with Glucantime® and/or cryotherapy in the treatment of zoonotic cutaneous leishmaniasis (ZCL) in patients referred to health centers of Isfahan, Golestan and Ilam Provinces of Iran as endemic foci of ZCL caused by Leishmania major besides Mashhad and Bam cities as endemic foci of anthroponotic cutaneous leishmaniasis (ACL) caused by with L. tropica.

Two hundred and seventy-eight patients with CL were included in the current study. All of the patients (100%) who received Nano Lip-AmB alone or in combination with Glucantime® and/or cryotherapy based on guideline of Iranian national committee for the treatment of CL. Two patients with 7 skin lesions, who was resident in ACL endemic area and received Nano Lip-AmB plus Glucantime® and another patient was a resident of ZCL endemic area and received Nano Lip-AmB plus cryotherapy showed clinical relapses after treatment.

Sina Ampholeish® in combination with other standard protocols of treatment of CL is well tolerated and with acceptable clinical efficacy rate.

Toxoplasma gondii infects nearly one-third of the world's population. Due to the significant side effects of current treatment options, identifying safe and effective therapies seems crucial. Nanoparticles (NPs) are new promising compounds in treating pathogenic organisms. Currently, no research has investigated the effects of zinc oxide NPs (ZnO-NPs) on Toxoplasma parasite. We aimed to investigate the therapeutic efficacy of ZnO-NPs against tachyzoite forms of T. gondii, RH strain in BALB/c mice.

In an experiment with 35 female BALB/c mice infected with T. gondii tachyzoites, colloidal ZnO-NPs at concentrations of 10, 20, and 50 ppm, as well as a 50 ppm ZnO solution and a control group, were orally administered four hours after inoculation and continued daily until the mices’ death. Survival rates were calculated and tachyzoite counts were evaluated in the peritoneal fluids of infected mice.

The administration of ZnO-NPs resulted in the reduction of tachyzoite counts in infected mice compared to both the ZnO-treated and control group (P<0.001). Intervention with ZnO-NPs significantly increased the survival time compared to the control group (6.2±0.28 days, P-value <0.05), additionally, the highest dose of ZnO-NPs (50 ppm) showed the highest mice survival time (8.7±0.42 days).

ZnO-NPs were effective in decreasing the number of tachyzoites and increasing mice survival time in vivo. Moreover, there were no significant differences in survival time between the untreated control group and the group treated with zinc oxide, suggesting that, bulk ZnO is not significantly effective in comparison with ZnONPs.

Recent studies have shown an increasing number of patients with cutaneous leishmaniasis (CL) who do not respond to pentavalent antimonials as the first line of treatment for CL. Nanocarriers such as extracellular vesicles (EVs) are efficient vehicles that might be used as drug delivery systems for the treatment of diseases. Therefore, we aimed to isolate and characterize the EVs of Leishmania major, load them with Amphotericin B (AmB), and investigate the toxicity and efficacy of the prepared drug form.

The EVs of L. major were isolated, characterized, and loaded with amphotericin B (AmB), and the EVs-Amphotericin B (EVs-AmB) form was synthesized. Relevant in vitro and in vivo methods were performed to evaluate the toxicity and efficacy of EVs-AmB compared to the control.

The anti-leishmanial activity of the EVs-AmB showed a higher percentage inhibition (PI%) (P = 0.023) compared to the AmB at different concentrations and time points. Obtained data showed a significant increase in the lesion size and parasite load in the lesion, PBS, and EVs mice groups in comparison with EVs-AmB, AmB, and Glucantime groups (P < 0.05), EVs-AmB had a significant decrease in lesion sizes in comparison with AmB (P < 0.05). Results showed that EVs-AmB decreased its toxicity to the kidneys and liver (P < 0.05).

EVs-AmB improved the efficacy of AmB in mouse skin lesions and reduced hepatorenal toxicity. Furthermore, EVs could be a promising nanoplatform for the delivery of AmB in CL caused by L. major.

We aimed to verify the susceptibility of Leishmania infantum, L. major and L. tropica, to commercial lectins in order to identify the three Leishmania species.

The degree of agglutination was determined both macroscopically and microscopically and was scored negative (-) to positive (from 1+- 4+) based on their percentage of agglutination.

Jacalin and UEA-1 were capable of agglutination of L. infantum isolates in both logarithmic and stationary phases at a concentration of 1000 μg/ml (100%). L. tropica isolates showed agglutination with the lectin UEA-1 in both logarithmic and stationary phases (62.5% and 87.5%). L. major and L. tropica showed 75% agglutination with lectin Jacalin in both logarithmic and stationary phases. L. tropica isolates showed 25% agglutination with the lectin WGA in the logarithmic phase. L. infantum, L. major and L. tropica isolates showed 25, 12.5 and 37.5% agglutination in the stationary phase, however, did not show agglutination in logarithmic phases. L. major isolates showed 12.5% agglutination with the lectin PHA in the stationary phase, however, were incapable of agglutination with the L. tropica and L. infantum in both logarithmic and stationary phases.

Despite the fact, that JCA and I-UEA lectins were not able to completely separate L. infantum, L. major and L. tropica. WGA lectin and PHA lectin can help in separating the species of Leishmania parasites.

Enzymatic digestion of extra cellular matrix proteins by proteinases of Leishmania promastigotes is a complex process. Hence, studies on functional proteomics of these enzymes can help select these enzymes as possible vaccine candidates or selecting candidates for chemotherapy and immunotherapy. Several proteolytic enzymes are involved in virulence of Leishmania spp. These enzymes are mostly serine, cysteine and metalloproteases. We aimed to detect proteases in Leishmania promastigote exosomes.

Serine, cysteine and metalloproteases were investigated in exosomes and lysate of L. major promastigote using gelatin zymography. The study was carried out in the Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran, in 2021.

Zymography findings of metalloproteinases showed transparent bands, including a 63-kDa glycoprotein (GP63). This glycoprotein is a major surface metalloproteinase. In addition, transparent bands belonged to serin proteases and cathepsin were demonstrated in gels associated to Leishmania promastigote lysate and exosomes.

Several metalloproteases, serin proteases and cathepsins were shown in promastigote lysate and exosomes of L. major, which could purified and used as fractions for immunodiagnostic.

Visceral leishmaniasis (VL) is one of the most important neglected tropical diseases. The zoonotic form of VL is endemic in some areas of Iran. We aimed to determine the status of VL identified in humans and canines in different parts of Iran from 2013 to 2022.

A national representative cross-sectional study was conducted in 10 provinces of Iran, including the national leishmaniasis reference lab. We employed the direct agglutination test (DAT) as a reliable serological method to detect anti-Leishmania infantum antibodies in humans and animal reservoir hosts. Additionally, a narrative literature review was conducted to identify relevant studies on VL seroprevalence in Iran from 2013 to 2023.

The results of 21281 human and 5610 canine serum samples from 2013 to 2022 are reported. Altogether, 448 (2.1%, 95%CI: 2.0-2.3) human serum samples showed anti-L. infantum antibody levels of ≥1:3200. Of these samples, 13716 (64.5%) were collected actively, which showed a seroprevalence of 0.6% (95% CI: 0.5-0.8) and 7565 (35.5%) were collected passively, which showed a seroprevalence of 4.8% (95%CI: 4.3-5.3). Overall, 1035 (20.1%, 95%CI: 19.0-21.2) of 5160 domestic dogs (Canis familiaris) samples showed anti-L. infantum antibody levels of ≥1:320. Northwest (2.8%) and northeast (0.96%) regions had the highest human VL seroprevalence, while northwest (21.5%) and south (14.4%) regions had the highest canine VL seroprevalence.

Zoonotic VL, an endemic parasitic disease, is still present in several different distinct areas across Iran. While human VL cases have shown a declining trend over the last decade, the prevalence of canine VL remains significant.

Toxoplasma infection is caused by Toxoplasma gondii, which is an intracellular protozoan parasite. This infection consequently lead various congenital disabilities during pregnancy in patients. Spiramycin (Spi), a macrolide antibiotic, is typically recommended for T. gondii infection in pregnant women. We aimed to prepare the nanoemulsion of spiramycin (NE-Spi) and to evaluate the activity of this formulation in tachyzoites of T. gondii, RH strain.

This study was conducted in 2019-2021 at the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. NE-Spi was prepared by spontaneous emulsification. The effects of this nanoemulsion on the viability of cultured cells were measured using MTT assay. To estimate the effects of NE-Spi on tachyzoites of T. gondii, RH strain, different concentrations of NE-Spi, S-Spi (suspension of spiramycin), and NE (nanoemulsion without any spiramycin) were added to tachyzoites and then stored for 30, 60, 90, 120 min and 24 h in 250 μg/ml concentration at room temperature. Finally, Tachyzoites mortality rates were evaluated by trypan blue staining. Of note, flow cytometry was conducted to confirm the obtained results.

The final particle size of NE-Spi was calculated to be 11.3 nm by DLS and TEM. Thereafter, using MTT assay, in 62.5 μg/ml concentration of NE-Spi, the Vero cells viability was obtained as 82%. The highest mortality rates of tachyzoites of T.gondii, RH strain were observed at 250 μg/ml concentration and after 120 min of exposure, but it was not significantly different from 24 h of exposure.

NE-Spi has lethal efficacy on T. gondii RH strain in-vitro.

In recent decades, effective herbal and natural compounds have been formulated with topical application, reduced side effects, and more economic benefits for Cutaneous Leishmaniasis (CL) treatment. Some of these include Propolis, Royal jelly, Ostrich oil, and Aloe Vera, which have widespread use in health products. The study aimed to evaluate the potential therapeutic effects of a prepared topical combination, including Propolis, Royal jelly, Ostrich oil, and Aloe Vera, on Leishmania major lesions under an in vivo model in BALB/c (concatenation of Bagg and Albino) mice. Fifty BALB/c mice were randomly divided into five groups of ten. CL lesions were induced by inoculation of metacyclic promastigotes at the base of the BALB/c mice's tails. The resulting lesions were treated topically with prepared natural combinations of Propolis, Royal jelly, Ostrich oil, and Aloe Vera. The lesion sizes were determined as the mean ± standard deviation. One-way analysis of variance (ANOVA) with the Bonferroni correction method was used for statistical analysis, with p < 0.05 considered statistically significant. Based on the data of this research, comparative ulcer size reductions were 63.4% (3.25 mm ± 0.24), 68.4% (3.55 mm ± 0.22), and 8.5% (0.44 mm ± 0.98) for the natural combination treatment 1, treatment 2, and Glucantime, respectively. Based on the results, the proposed natural combinations were significantly more effective at reducing ulcer size than Glucantime, the standard drug (p<0.05). Collectively, we conclude that a combination consisting of Propolis, Royal jelly, Aloe Vera, and ostrich oil is effective in treating CL in mice and may provide a new agent in the treatment of Leishmaniasis.

Cutaneous leishmaniasis (CL) is an endemic infection in the Middle East, including Iran that is also spreading to new foci. We aimed to determine the leishmaniasis species causing CL in Alborz province.

Overall, out of 55-suspected CL patients referred to health centers in Alborz Province, north central Iran in 2019, 40 patients had positive smear for CL based on optical microscopy. The internal transcribed spacer 1 (ITS1) of nuclear ribosomal DNA (rDNA) was amplified by PCR. Leishmania species were identified by PCR–restriction fragment length polymorphism (PCR-RFLP) using BshF I (Hae III) enzyme.

Out of the 40 positive patients with CL, 34 cases (85%) had been caused by Leishmania (L) major and six (15%) by L. tropica. Fifteen patients had no history of traveling to the disease endemic areas, of which nine were Iranians. Skin lesions and scars caused by CL were mostly observed on the hands and face. Moreover, more than two skin lesions were observed in 22 cases (55%), all of which were infected with L. major. A single skin ulcer was seen in 18 (45%) of the CL patients.

Climate change, reduced rainfall, and demographic changes such as migration into Alborz Province and the increasing marginalization of the population and their entry to settle in new areas might have caused natural transmission of both L. tropica and L. major in this province.

Leishmaniasis is currently considered a re-emerging or emerging infection based on the geographic region. The outcome of leishmaniasis vastly depends on Leishmaniahost interaction. This preliminary study aimed to show the association of human leukocyte antigen (HLA) class I and II genes with healed and non-healed cutaneous leishmaniasis (CL), and symptomatic and asymptomatic visceral leishmaniasis (VL) compared with control groups in Iran.

Ninety-five people, including 31 patients versus 64 individuals in the control group, were enrolled. Among them, 20 patients had confirmed CL based on amastigote observation, 10 had improved CL and 10 non-healed CL. Eleven patients were suffering from confirmed VL based on direct agglutination test (Five asymptomatic and six symptomatic VL cases). Besides, they were residents in an endemic area of VL in the northwest of Iran. To select a control group, it was ensured that they had no history of leishmaniasis. Peripheral blood samples were collected from each patient. After DNA extraction, HLA typing was conducted using polymerase chain reaction - sequence-specific priming (PCR-SSP). Subsequently, data were statistically analyzed by SPSS.

There was a statistical relationship between the presence of HLA-A26 and CL, healed CL and the existence of the B38 allele, C1 allele and symptomatic VL, as well as B1.4 allele and asymptomatic VL (P˂0.05).

This primary finding indicates that several HLA genes have a potential role in the susceptibility of Iranian people to CL and VL.

The primary aim of this study is to determine infection to Leishmania parasites in the wild population of Phlebotomus caucasicus and Phlebotomus mongolensis using molecular methods in some important zoonotic cutaneous leishmaniasis foci in Iran.

Sand flies were collected from active colonies of rodent burrows from 16 trapping sites using sticky trap pa per. In order to detect and identify of Leishmania parasites in females Ph. caucasicus and Ph. mongolensis, the Nested–PCR amplification of ITS2-rDNA region was performed to generate amplicon with 245bp for Leishmania major, 206bp for L. gerbilli and 141bp for L. turanica.

In the current study we found DNA of different gerbil parasites such as L. major and L. turanica, and mixed infection of L. major/L. turanica in Ph. caucasicus and Ph. mongolensis. It should be noted that, in Iran, natural infec tion with Leishmania parasites is recorded for the first time in this study in Ph. mongolensis.

Both species of Ph. caucasicus and Ph. mongolensis not only may participate in the ZCL transmission cycle between reservoir hosts, but also results of this study support the  role of these species as secondary vectors in the transmission of leishmaniasis to humans.

Fleas (Insecta: Siphonaptera) are considered as highly specialized bloodsucking on mammals such as dogs. The existence of three factors, namely a vast distribution area, different hosts, and digestive system with a specific mecha nism for digesting blood has led to species of fleas who nourish from mammals be introduced as the potential vectors of diseases. The aim of this study was to assess Leishmania infantum natural infection of dog fleas in northwest Iran in 2018.

A total of 20 infested domestic dogs (Canis lupus familiaris) were randomly selected from 5 villages. Fleas were collected using brushing against dog hairs and fine forceps. Then, they were morphologically identified and pre served in ethanol for molecular assay. The kinetoplast DNA of the parasite was used for detection of Leishmania infan tum using a semi-nested polymerase chain reaction (PCR) assay.

The human flea, Pulex irritans, and the cat flea, Ctenocephalides felis were identified on 40% and 35% of dogs, respectively. The results of PCR indicated that L. infantum was found in the Ctenocephalides canis (75%) and C. felis (66.7%) col lected from infected dogs. No leishmanial infection was observed in P. irritans.

It is concluded that fleas could be infected by Leishmania infantum, but maintenance of the parasite and their vectorial competence needs to be determined.

Visceral leishmaniasis (VL) also known as Kala-azar is considered as one of the zoonotic infections in Mediterranean countries. The reservoir of this infection is dogs and sand flies are considered as its vectors. Due to reported sporadic cases of Kala-azar in the past five years in Shahroud County, Semnan Province, Iran, this study aimed to investigate the status of this infection in this area and to determine its seroepidemiology to take required measurements for infection control and treatment.

This descriptive cross-sectional study was conducted on 504 subjects residing in seven villages in Shahroud County, Semnan, Iran. Samples were randomly selected using the cluster sampling method and blood samples were collected from subjects aged up to 13 years old (90%) and adults over 13 years old (10%) from September 2018 to May 2019. After separating sera from whole blood, samples were subjected to direct agglutination test (DAT) to detect anti-Leishmania infantum antibodies. First dilutions were prepared from 1:10 to 1:800 for human samples.

Results of serological testing with 1:800 titration indicated that no sample was positive for antibody against Leishmania infantum. After secondary screening, 10 cases (1.98%) showed the antibody titer of 1:100, while four cases (0.79%) showed the antibody titer of 1:400. According to clinical findings, no patient was suffering from fever, weight loss, splenomegaly, hepatomegaly, and cachexia and therefore did not show the symptoms of Kala-azar.

The results of the current study indicate that Kala-azar is not prevalent in Shahroud County.

After the earthquake in 2017 a few new cases of visceral leishmaniasis (VL) were reported from SarPol-e-Zahab district of Kermanshah Province, western part of Iran. This study was conducted to determine the seroprevalence in Kermanshah Province.

This descriptive cross-sectional study was conducted on children up to 12 years of age from SarPol-e-Zahab County, Kermanshah Province, western part of Iran in 2021. For each individual, a questionnaire including age, sex, clinical features, history of the disease, and contact with canines as reservoir hosts of VL were completed, separately. To determine VL seroprevalence, blood samples were collected from the children and after centrifugation, the sera samples were separated and tested using Direct Agglutination Test (DAT) for detection of anti-L. infantum antibodies. Statistical analyses were performed using SPSS16.

Totally, 13 persons were seropositive; 7 samples with titer 1:800, 3 samples had 1:1600, 2 samples had 1:3200 and 1 sample had 1:6400. None of the seropositive cases had a history of kala-azar. There was no significant difference between males and females at titers of anti-Leishmania specific antibodies.

L. infantum infection is being circulated with low prevalence in children up to 12 years old from SarPol-e-Zahab County but it is necessary that the surveillance system is regularly monitored among physicians and public health managers in the studied areas.

Various arthropods, including Rhipicephalus sanguineus and Ctenocephalides felis felis have been sug-gested as secondary vectors of Leishmania spp. many years ago. This study was conducted to determine zoonotic cuta-neous leishmaniasis (ZCL) PCR positivity of reservoir hosts and their ectoparasites for Leishmania spp. in Segzi plain in Esfahan Province from October 2016 to October 2017.

Microscopic examination and nested PCR were used to detect and identify Leishmania spp. isolated from rodents' ears and ectoparasites, and then, the results were confirmed by two methods, PCR-restriction fragment length polymorphism and sequencing.

Totally, 93 rodents (92 Rhombomys opimus and one Nesokia indica) and nine different species of ectoparasites (n=527) including fleas, mites, and ticks were collected during different seasons in the study area. Fourteen R. opimus were positive for Leishmania spp. by microscopic examination while one N. indica and 77 R. opimus were positive by nested PCR. The infection rate of rodents with Leishmania major and Leishmania turanica was 39.79% (n=37) and 15.05% (n=14), respectively. Mixed natural infections with L. major and L. turanica were seen in rodents. Moreover, 72.22% of fleas (39/54), 75.0% of mites (5/8), and 100% of tick nymph (1/1) were PCR positive for Leishmania parasites.

The highest rate of infection with L. major and L. turanica in R. opimus populations was observed in summer and spring, respectively. It is suggested that the role of L. turanica and the probable role of ectoparasites in the epidemiology of disease should be investigated. A Xenodiagnostic test is recommended for future study.

Pentavalent antimonial has been a drug of choice against leishmaniasis, despite the emergence of treatment failure. Identification of resistance markers is urgently needed to design new therapeutic strategies. Iron-Superoxide dismutases (Fe-SODs) are antioxidant enzymes contributing to detoxify reactive oxygen species to prevent a cell from oxidative stress. Since antimonial compounds induce oxidative stress, in this survey, the expression of SOD genes was investigated to identify their expression pattern in clinical resistant isolates.

This cross-sectional survey was done in Mashhad City, northeast of Iran during 2014 to 2019. The RNA expression level of mitochondrial (SODA) and glycosomal (SODB) superoxide dismutase was investigated in 25 antimony responsive (n=15) and unresponsive (n=10) anthroponotic cutaneous leishmaniasis (ACL) patients. Total RNA extraction and cDNA synthesis, the qRT-PCR approach was utilized to investigate the relative RNA expression level.

The transcript level of SODs was over-expressed in the most resistant isolates. Gene expression analysis demonstrated the over-expression of SODA and B by a factor of 3.8 and 4.81, respectively, in resistance isolates vs. sensitive ones.

Aberrant expression of SODA/B in unresponsive parasites could potentially implicate in detoxifying antimony-induced oxidative stress. Moreover, SODs might be considered as potential predictive markers of the response to antimonials in ACL patients in endemic areas.

Infections by Plasmodium falciparum, are becoming increasingly difficult to treat. Therefore, there is an urgent need for novel antimalarial agents’ discovery against infection. In present study, we described a 2’-O-Methyl gapmer phosphorothioate oligonucleotide antisense targeting translation initiation region of 3D7 strain RH5 gene.

The study was conducted in Pasteur Institute of Iran in 2020. ODNs effects were measured by microscopic examination and real time RT-PCR. For microscopy, microplates were charged with 2’-OMe ODNs at different dilutions. Unsynchronized parasites were added to a total of 0.4 ml (0.4% parasitemia, 5% red blood cells), and slides were prepared. Proportion of infected cells was measured by counting at least 500 red blood cells.

RH5 genes start codon regions selected as conserved region besed on alignment results. Gap-RH5-As which was complementary to sequence surrounding AUG RH5 start codon significantly reduced parasite growth (>90% at 50 nM) compared to sense sequence control (Gap-RH5-Se) (17%), (P<0.001). RH5 transcripts were dramatically reduced after exposed to ODNs at a concentration of 5-500 nM for 48 h.

Gemnosis delivery of a chimeric gapmer PS-ODN with 2’-OMe modifications at both sides had high antisense activity at low concentrations (10-100 nM) and shown a good efficiency to reach to target mRNA in human RBCs. Anti-parasite effect was correlated to reduction of target gene mRNA level. In addition, 2’-OMe ODNs free delivery is an effective way and does not need any carrier molecules or particles.

A variety of haemoprotozoa including Plasmodium, Haemoproteus and Leucocytozoon cause infections in birds and are transmitted by some known vectors. These parasites cause anemia, low appetite, weakness and ultimate ly death in birds. The present study was aimed to determine these parasites, in birds of Mazandaran and Golestan prov inces in Iran.

The project was performed on 340 live birds in 2016. The samples were collected from February to Septem ber 2016, from each bird, two thin and thick blood smears were prepared and the remaining blood about 1ml was kept in EDTA-containing tubes for molecular studies. The slides were stained with 10% Giemsa, then examined microscopical ly. About ten percent of the negative samples were considered for Polymerase Chain Reaction (PCR) technique, using specific primers to diagnose Plasmodium and Haemoproteus spp. Electrophoresis was done for PCR products and rele vant bands to the parasites were identified based on the size. The considered birds belonged to ducks, chickens, roosters, and pigeons.

From 340 microscopically examined blood samples 32 (9.5%) samples were positive. Twenty-five (7.35%) of them were infected with the genus Haemoproteus. Seven samples (14%) out of 50 microscopically negative samples were found as Haemoproteus or Plasmodium spp when PCR technique was employed.

This study revealed the existence of malaria parasites and other haemosporidia in birds in Iran. Employing molecular methods (PCR examination) could detect more infections.

Detection of Leishmania RNA virus (LRV) in Old World Leishmania species and their possible role in the disease prognosis requires sensitive and specific methods, preferably independent of the viral genome. We aimed to develop an indirect immunofluorescence antibody (IFA) assay to detect LRV in the Old World Leishmania parasites.

Clinical samples were collected from 86 cutaneous leishmaniasis (CL) patients in different endemic areas of CL in Iran, during 2017-2019. For antibody preparation, the viruses were obtained from sediment of an LRV-infected L. major culture-using freeze and thaw cycles followed by gradient cesium chloride centrifugation. The purified viruses were used to immunize a male 3-4 months rabbit. Various dilutions of the LRV-immunized rabbit's serum and a conjugated antibody were deployed to detect LRV in 48 isolates by IFA assay.

LRV virus was detected in four of the 48 CL cases using IFA method. Amplification of a partial fragment of RNA-dependent RNA polymerase (RdRp) gene from the isolates confirmed the IFA results. In phylogeny, the generated RdRp sequences from four isolates were grouped with the other Old World LRVs, but separate from L. aethiopica LRVs, which appeared as a highly supported distinct clade.

Further optimization of this approach to detect the LRV directly in lesion scrapings can make it a more reliable tool for field studies and disclosing the virus's possible role in disseminating and unusual clinical features.