Phylogenetic analysis and antimony resistance of Leishmania major isolated from humans and rodents.

Cutaneous Leishmaniasis (CL) is one of the world's neglected diseases which is caused by Leishmania spp. The aim of this study was to assess molecular profile and antimony resistance of Leishmania isolated from human and rodent hosts. Samples were collected from suspected CL patients referred to health centres and wild rodent's traps in Gonbad-e-Qabus region, north-eastern Iran. Smears were subjected to PCR-RFLP to identify Leishmania species. In addition, ITS1-PCR products were sequenced for phylogenetic analysis. Clinical isolates and rodent samples were subjected to MTT assay to determine IC50 values and in vitro susceptibilities. Expression levels of antimony resistance-related genes were determined in CL isolates. Out of 1,949 suspected patients with CL and 148 rodents, 1,704 (87.4%) and 6 (4.05%) were positive with direct smear, respectively. Digestion patterns of BusRI (HaeIII) endonuclease enzyme were similar to what expected for Leishmania major. Phylogenetic analysis revealed that the highest interspecies similarity was found between current L. major sequences with L. major obtained from Russia and Uzbekistan. Out of 20 L. major samples tested, 13 (65%) were resistant to meglumine antimoniate (MA) treatment, with an activity index (AI) exceeding 4. The remaining 7 samples (35%) responded to MA treatment and were classified as sensitive isolates, with a confirmed sensitive phenotype based on their AI values. The comparison expression analysis of three major antimony resistance-associated genes in unresponsive clinical isolates demonstrated significant fold changes for TDR1 (4.78-fold), AQP1 (1.3-fold), and γ-GCS (1.17-fold) genes (P < 0.05). Herein, we demonstrate genetic diversity and antimony resistance of L. major isolated from human and reservoir hosts in north-eastern Iran, which could be the basis for planning future control strategies.

Is There Any Relationship between Trichomonas vaginalis Infection and Male Urethritis Risk? A Systematic Review and Meta-Analysis.

Background: Male urethritis is one of the most common genital tract syndromes. Though the number of patients with urethritis is increasing worldwide, the cause of many cases of non-gonococcal urethritis (NGU) is still unknown. Objectives: This study aimed to delineate the association between Trichomonas vaginalis (T. vaginalis) infection and male urethritis. Methods: The literature was searched in PubMed, Scopus, and Web of Science databases using the search terms "urethritis," "Trichomonas vaginalis," "trichomoniasis," and "male urethritis" up to February 2020. Overall risk difference(RD) was applied to assess the relationship between T. vaginalis infection and male urethritis. Results: In total, seven articles were included in this systematic review and meta-analysis study. Our meta-analysis involved the review of case-control studies, including 2,242 urethritis cases and 929 individuals as controls. Among subjects examined for trichomoniasis, in the case group, 211 males were infected, and in the control group, 32 individuals were infected. The overall risk difference (RD) was 0.06, and the total reported p value was 0.00001. Although the result of our meta-analysis was not significant, it was shown that the risk of urethritis is 0.06 more in trichomoniasis patients than in the non-exposed group. Conclusion: Findings from the included papers showed that trichomoniasis is not a risk factor for male urethritis. Although trichomoniasis alone is not the main cause of urethritis, it can be considered one of the risk factors in male urethritis. Therefore, in the future, it is necessary to perform further studies to clarify the detailed association between T. vaginalis infection and urethritis risk in male patients.

Molecular Evidence of Emerged Pulmonary Lophomoniasis due to Lophomonas blattarum among Hospitalized Patients in Southwestern Iran: A National Registry-Based Study.

Objectives: Lophomonas protozoan is an emerging pathogen transmitted through arthropods such as cockroaches. Lophomoniasis is still a mysterious disease with many unknown epidemiological aspects. The current study aimed to determine the prevalence of lophomoniasis among patients who were hospitalized in Hajar Hospital, Shahrekord, southwestern Iran, using a conventional PCR technique. Methods: In this retrospective study, 132 frozen bronchoalveolar lavage fluid (BALF) specimens from patients with respiratory disorders hospitalized in Hajar Hospital, Shahrekord district, southwestern Iran, were analyzed during 2020-2021. Samples are referred to the Iranian National Registry Center for Lophomoniasis (INRCL), Mazandaran Province, Northern Iran, for detecting Lophomonas spp. infection by a conventionally small subunit ribosomal RNA (SSU rRNA) PCR test. Results: A total of 132 frozen BALF specimens were examined, 36 (27.3%) tested Lophomonas spp. positive using the conventional PCR technique. Also, based on sequencing data and blast analysis, the presence of L. blattarum species was confirmed. The average age of Lophomonas spp.- positive patients was 67.02 ± 15.14 years. Out of the 36 positive subjects, 63.9% were male and 36.1% female. Male and Lophomonas infection had a significant correlation (p=0.001). Our findings revealed that L. blattarum infected nonsmokers more than smokers (p=0.001). The most common underlying disease was also bronchitis. Conclusion: Our results showed, for the first time, that pulmonary lophomoniasis caused by L. blattarum is a common and emerging disease in the study area, southwestern Iran. Furthermore, our findings support the use of the PCR test to detect Lophomonas infection in archived frozen clinical samples.

Global Status of Emerging Lophomonas Infection: A Systematic Review of Reported Cases (1993-2020).

Purpose: Lophomonas spp., is an emerging protozoan parasite that belongs to the Parabasalids (Parabasalia, lophomonadida) which infects the respiratory tracts of humans. Despite the presence of a few reports of human lophomoniasis, the true burden of Lophomonas infection is unknown. This systematic review aimed to elucidate the latest global status of publications reporting human cases of lophomoniasis as a new emerging protozoal disease. Methods: A comprehensive and systematic search was performed in 10 (five English and five Persian) databases for studies reporting cases of lophomoniasis between 1993 and March 2020 (27 years). Then, the selected articles were carefully reviewed and screened based on the inclusion and exclusion criteria. Results: Overall, 32 eligible publications reporting 307 lophomoniasis cases from around the world are included in this review. The patients were aged between 1 month and 84 (mean age = 23.7 years). The highest number of cases (n = 171; 55.7%, P < 0.04) significantly belonged to the juvenile age group (aged ≤18 years). The male to female ratio of the cases was almost equal, and no statistically significant difference between them was observed. The maximum number of cases (n = 237) was reported from Iran. Most cases (n = 196; 63.85%) had no history of underlying diseases/organ transplantation (P < 0.001). Moreover, the BAL specimen was the most commonly used clinical sample to diagnose lophomoniasis (P < 0.001). Conclusion: Our findings reveal that the prevalence of lophomoniasis is likely to be markedly underestimated when evaluated based on published case reports. Additionally, our data, at least for the time being, supports the idea that Lophomonas spp. should not be considered as an opportunistic infection. Thus, current work sheds light on some controversial issues regarding the epidemiological aspects of lophomoniasis.

A Convenient and Sensitive kDNA-PCR for Screening of Leishmania infantum Latent Infection Among Blood Donors in a Highly Endemic Focus, Northwestern Iran.

BACKGROUND: Recent global evidences showed that asymptomatic blood donor carriers of Leishmania infection will appear as a threat for blood transfusions recipients in endemic areas. As yet, there is no appropriate diagnostic procedure for detecting infection of blood donors in blood banks. SUBJECTS AND METHODS: The present study was aimed to apply various current diagnostic tests among blood donors in an endemic area of visceral leishmaniasis (VL), Ardabil Province, northwestern Iran. Blood samples were gathered from 860 blood donors in endemic areas of the province between 2017 and 2018, at eight blood donation centers. These samples was assessed using microculture, serological (DAT and rK39-ICT) and molecular based (conventional kDNA-PCR and HRM-PCR) tests. RESULTS: Of 860 eligible donors, 24 (2.8%) were seropositive for VL by DAT, and 388 (45%) were positive by kDNA-PCR. Moreover, 19 (19/860) were positive for both of them. Out of 19 subjects, 5.3% (1/19) was positive by rK39-ICT, 10.5% (2/19), and 79% (15/19) were detected positive in microculture and HRM-PCR methods, respectively. Nineteen donors were followed up for 2 years, of which 16 (84.2%) had a serological conversion, and 4 (21%) were positive by kDNA-PCR. The sensitivity of kDNA-PCR, and HRM-PCR procedures in detecting Leishmania parasite was found to be 98.7%, and 79%, respectively. CONCLUSIONS: Our findings justify the use of kDNA-PCR as a convenient and sensitive tool for screening subjects with leishmanial latent infection in blood banks at least in endemic regions. In these areas, however, a PCR-based test should be used to validate Leishmania infection among seropositive donors.

In vitro anti-Acanthamoeba activity of the commercial chitosan and nano-chitosan against pathogenic Acanthamoeba genotype T4.

Acanthamoeba keratitis (AK) is a rare but serious infection of the eye and can lead to blindness. The effective and safe medical therapy remains unclear for AK until present. Antimicrobial activity and biological characteristic of chitosan encourage screening of it against Acanthamoeba. Thus, in vitro anti-amoebic activities of commercial chitosan and nano-chitosan were tested on pathogenic Acanthamoeba genotype T4, a causative agent of human AK. The Acanthamoeba spp. was isolated from the keratitis patient. The Acanthamoeba genotype T4 was approved using PCR method followed by sequencing technique. Chitosan nanoparticles was prepared using ionic gelation method and characterized by their physicochemical properties. In the present study, the in vitro activity of serial dilutions (12.5, 25, 50, 100, and 200 µL/mL) of commercial chitosan and nano-chitosan were evaluated against Acanthamoeba trophozoites and cysts. The finding of nano-chitosan particle size by DLS was 118 nm with a PDI of about 0.134. Zeta potential value was found to be 42.7 mV. The obtained results showed that the tested chitosan and nano-chitosan presented anti-amoebic activities dependent to time and concentration. The inhibitory effect of the chitosan and nano-chitosan is enhanced by increasing the concentration and incubation time. The inhibitory effect of nano-chitosan on both trophozoites and cyst was more than chitosan. According to the results, nano-chitosan shows the potent activity against Acanthamoeba T4 and could be used for the development of novel and safe therapeutic approaches in the future.

Phylogenetic position of Leishmania tropica isolates from an old endemic focus in south-eastern Iran; relying on atypical cutaneous leishmaniasis.

Cutaneous leishmaniasis (CL) is a major health problem in Iran, with a heavy burden on human health and society. There is little knowledge about the molecular epidemiology of the disease, as well as phylogenetic relationship of causative agents in south-eastern Iran. The aim of the present study was to investigate the molecular aspects of CL, especially atypical CL in the Bam district, Kerman province, south-eastern Iran, as an endemic region of CL in Iran. The smears were collected from lesion samples of 353 patients clinically suspected to CL, who attended local health centres in the Bam district during 2016-2017. Direct smears were examined for Leishmania parasites using the Giemsa staining technique. Amplification of kinetoplast DNA (kDNA) and the ribosomal internal transcribed spacer 1(ITS-1) gene were carried out using polymerase chain reaction (PCR). Then, the ITS1-PCR products were sequenced for phylogenetic analysis. Overall, 278 cases were confirmed as CL by microscopic examination of Giemsa-stained slides. Clinical presentation of the lesions was basically of two types: (a) typical lesions and (b) atypical including lupoid ulcers, sporotrichoid, nodular and exudative lesions. The PCR assay on all specimens of skin lesions proved L. tropica as the main pathogenic agent. Phylogenic analysis revealed high similarity among isolates from the Bam district in the south-east with isolates from Birjand in eastern Iran, as well as with isolates from Herat province in western Afghanistan. The study provided valuable information concerning the genetic diversity of the parasite as one of the factors influencing the clinical manifestations in CL in south-eastern Iran, which could be the basis for planning future control strategies.

Presence and diversity of Leishmania RNA virus in an old zoonotic cutaneous leishmaniasis focus, northeastern Iran: haplotype and phylogenetic based approach.

OBJECTIVE: Leishmania RNA virus (LRV) is a double-stranded RNA (dsRNA) virus that circulates within many species of the Leishmania parasite. In this study, we aimed to investigate the presence of LRV2 circulating in Leishmania isolates in an old focus of ZCL located in northeastern of Iran. METHODS: Leishmania isolates were collected from 85 patients that confirmed to have cutaneous leishmaniasis (CL) based on parasitological examination. To identify the Leishmania isolates, species-specific primer sets were applied for molecular identification. The presence of LRV2 was performed by RdRp-semi nested-PCR. The genetic diversity were calculated using MEGA and DnaSP. To assess haplotype diversity, 31 LRV2 strains in different regions were surveyed using analysis a 292-bp section of the RdRp sequences. RESULTS: Out of 85 patients, 83 (97.6 %) were diagnosed with L. major and 2 (2.4 %) with L. tropica. LRV2 virus was detected in 59 (69.4%) of the CL cases. For the first time, LRV2 was reported in one L. tropica strain in Iran. The current LRV2 sequences indicated the highest similarities to an Old World LRV2. Moreover, 10 unique haplotypes were identified based on the analyzed sequences of the RdRp gene. CONCLUSIONS: Our results indicated the highest occurrence of Leishmania/LRV2 co-circulation in this known ZCL focus from northeastern Iran. Phylogenetic analyses of LRV2 sequences confirmed that these isolates belong to the order of LRV2 from the Old World. This study offered an insight into LRV2 haplotype that the informative issue can be used for genetic research of LRV2 in other regions.

Actin Gene-Based Molecular Typing of Trichomonas vaginalis Clinical Isolates from the North of Iran.

AIM: The aim of the current study is to evaluate the prevalence of trichomoniasis in men and women in the north of Iran and to find genotypes in the positive clinical specimens based on T. vaginalis actin gene. MATERIALS AND METHODS: Women's genital (n = 500) and men's urine (n = 1500) samples were collected from the participants referred to clinics in Mazandaran Province, northern Iran, during 2006-2018. In addition, 1500 Pap smear specimens, archived in the Bu Ali Hospital, Sari City, Mazandaran Province, northern Iran, were examined. The specimens were examined based on parasitological methods, nested polymerase chain reaction (PCR), PCR-restriction fragment length polymorphism, and phylogenetic analysis. RESULTS: Overall, 17 (0.48%) of 3500 specimens were positive by PCR. Total prevalence was 0.55% (n = 2000) for women, of which 500 (1.4%; n = 7) specimens were collected freshly, and 1500 (0.26%; n = 4) were Pap smears. Moreover, six (0.4%) out of 1500 men urine specimens were positive. Overall, genotypes G, E, and I were detected with the prevalence of seven (0.2%), seven (0.2%), and three (0.08%), respectively. There was no significant statistical difference among the prevalence of the detected genotypes (P > 0.05). CONCLUSION: As a whole, the prevalence of trichomoniasis was low in the studied area in the north of Iran and, most importantly, the genotypes of E, G, and I were distributed among men and women in the province.

The global status of Dirofilaria immitis in dogs: a systematic review and meta-analysis based on published articles.

Dirofilaria immitis is a parasitic filarial nematode responsible for heartworm disease in domestic as well as wild canines and felines and pulmonary or cutaneous infections in humans. This systematic review and meta-analysis aimed to evaluate the status of D. immitis in dogs based on available literature. Four English language databases (ISI Web of Science, Scopus, PubMed and Science Direct) containing data on D. immitis prevalence in dogs were thoroughly searched resulting in the inclusion of 193 studies. The findings revealed that the pooled and weighted prevalence of D. immitis infection in dog population based on published papers throughout the world was 10.91% (95% CI=10.18-11.65). In addition, subgroup analysis based on meta-regression revealed that a significant difference between the pooled and weighted prevalence of D. immitis in dogs and country (ß =0.14, P=0.049). Given the relatively high prevalence of D. immitis infection in dogs and its adverse effects, it is suggested that to perform more research on the prevention and control of dirofilariasis infection in dogs worldwide.

HRM-PCR is an accurate and sensitive technique for the diagnosis of cutaneous leishmaniasis as compared with conventional PCR.

BACKGROUND: Cutaneous leishmaniasis (CL) is considered one of the main health problems in Iran. Therefore, it is required for control and therapeutic purposes, an accurate and fast tool for the diagnosis and identification of Leishmania species. METHODS: In the present study, three techniques, including microscopic examination, conventional PCR, and high-resolution melting (HRM)-PCR, have been evaluated, to find the most accurate and rapid test. In total, 105 skin scraping smears were taken from suspected dermal lesions of patients belonging to two known endemic CL areas, Gonbad and Bam districts, in Iran. Subsequently, the specimens were analyzed with microscopic, conventional PCR, and HRM-PCR techniques. RESULTS: Most positive samples (89.5%) were observed using HRM-PCR, and among the three techniques, HRM-PCR was the most sensitive (89%, 95% CI 81-94) technique. Microscopic examination test had the lowest sensitivity (57%, 95% CI 47-66%). The highest agreement among positive samples was observed between HRM-PCR and conventional PCR tests. DISCUSSION: Our results showed that the HRM-PCR technique is the most accurate and sensitive test for recognizing CL, and also a valuable alternative test for conventional PCR to detect various species.

Frequency and genetic diversity of Blastocystis subtypes among patients attending to health centers in Mazandaran, northern Iran.

Blastocystis hominis is a common intestinal protozoan with global distribution. The prevalence of B. hominis is extremely diverse in different countries. Molecular studies show that there is a great deal of genetic variation in the Blastocystis parasite isolated from humans and animals. Therefore, this study was conducted to determine B. hominis genotypes in patients attending to health centers of Sari, Mazandaran Province, north of Iran. 420 fecal specimens were collected from individuals in health centers of Sari from April to December 2017. The samples were examined by direct methods, parasite test (formalin ether) and Giemsa staining. In order to determine the genotypes of Blastocystis parasite, all positive samples in direct methods were tested by PCR using SSU rDNA gene. Eventually, seven positive molecular samples were sequenced and recorded in the gene bank. The phylogenetic analysis was done by drawing a gene tree using the neighbor-joining procedure. B. hominis was detected in 60 stool specimens by microscopic method. All these 60 samples were positive by PCR and the bands 310 bp were observed. According to the phylogenetic tree, it was revealed that B. hominis isolates of Mazandaran Province are more similar to subtype ST3 (ST type) that reported in Khorramabad in Lorestan Province (West of Iran). The present study showed that, like most studies, ST3 is the dominant genotype of protozoan in Sari. Improvement of public health awareness, relative improvement in environmental health, urban wastewater treatment, proper clothing coverage and the use of gloves during dealing with livestock and the use of domestic water treatment devices can reduce the Blastocystis infection in this area.

First Molecular Evidences of Acanthamoeba T3, T4 and T5 Genotypes in Hemodialysis Units in Iran.

BACKGROUND: Acanthamoeba is a genus of the free-living amoeba that is widespread in the environment and is a causative agent of opportunistic infections in human. This study aimed to investigate the existence and genotyping of Acanthamoeba species in hemodialysis units in Iran. METHODS: In the present study, forty water samples of hydraulic systems and twenty dust samples were collected from two hemodialysis units in Mazandaran Province, northern Iran. The samples were cultivated on non-nutrient agar and genotyping was performed by targeting the 18S rRNA gene. RESULTS: Both morphology and molecular analyses showed that 17.5% (7/40) of water samples and 50% (10/20) of dust samples were positive for Acanthamoeba spp. The sequencing analysis of these isolates was found to be T3, T4 and T5 genotypes. DISCUSSION: To the best of our knowledge, this is the first investigation to identify of Acanthamoeba species in hydraulic system of hemodialysis units in Iran. High contamination of hemodialysis units with virulent T4 genotype of Acanthamoeba may poses a risk for biofilm formation. Our results support urgent need to improve filtration methods in dialysis units and monitoring hemodialysis patients for Acanthamoeba infections.

Medicinal Plants with Anti-Trichomonas vaginalis Activity in Iran: A Systematic Review.

BACKGROUND: Trichomoniasis, due to Trichomonas vaginalis, is one of the most common sexually transmitted parasitic diseases in the world such as Iran. This systematic review aimed to explore the studies evaluating the medicinal herbs with anti-T. vaginalis activity which used in Iran. METHODS: Articles published in 4 Persian and 4 English databases were obtained between 2000 and 2015 including Google Scholar, PubMed, Science Direct, Scopus, Magiran, Barakatkns (formerly IranMedex), Elm net, and SID (Scientific Information Database). Studies out of Iran, studies on animal models and articles on other parasite species than T. vaginalis were excluded from this review. RESULTS: Twenty-one articles including in vitro experiments, met our eligibility criteria. Thoroughly, 26 types of plants were examined against T. vaginalis. Medicinal herbs such as Artemisia, Zataria multiflora, and Lavandula angustifolia are remarkably effective on T. vaginalis. As such, use of other parts of these plants in different concentrations and timelines is recommended for future in vivo studies. CONCLUSION: The present systematic review provides comprehensive and useful information about Iranian medicinal plants with anti-T. vaginalis activity, which would be examined in the future experimental and clinical trials and herbal combination therapy.

Human urogenital myiasis: A systematic review of reported cases from 1975 to 2017.

The public health importance of myiasis [infestation with dipterous (fly) larvae] remains unknown. This disease is spread worldwide in animals and humans, but baseline data on its prevalence are limited. In particular, knowledge on human urogenital myiasis (UGM) is scattered. As such, a systematic search was undertaken of five English and five Persian databases for publications describing UGM cases in English or Persian published between 1975 and 2017. In total, 45 papers reporting 59 UGM cases from various regions of the world are included in this review. All included papers were from the English databases. The age of patients ranged from 5 to 89 years, and the mean age was 40.6 years. Thirty-six of the patients were female and 19 were male. The highest number of cases (n = 12) was reported from Brazil. The most common genera causing UGM were Psychoda spp. (23.7%) and Cochliomyia spp. (11.8%). The vagina was the most commonly reported anatomical location of UGM for women, and the urogenital tract was the most commonly reported location for men. Thirteen cases were reported from rural areas and eight cases from urban areas; the location of other cases was not specified. The incidence of UGM is likely to be substantially underestimated when evaluated based on published case reports. Epidemiological studies, such as questionnaires to medical doctors, could help to gather the necessary baseline data on the occurrence of UGM.

Genetic diversity at the C-terminal domain of knob-associated histidine-rich protein (KAHRP) of Plasmodium falciparum isolates from Burundi, Eastern Africa.

The knob-associated histidine-rich protein (KAHRP) is an exported parasite protein and the major component of infected erythrocytes by Plasmodium falciparum. P. falciparum histidine-rich protein-1 (PfHRP-1) is docked by KAHRP, which this interaction plays a significant role in cytoadherence of the malaria protozoan to erythrocytes and pathogenicity. The most polymorphic region of the PfHRP-1 is the C-terminal of decapeptide repeat domain (region III). The main objective of this study was to explore the genetic diversity at the region III of KAHRP in P. falciparum isolates from Burundi. In the present study, the nested PCR was performed for the amplification of the coding gene (kahrp gene) for region III in 35 P. falciparum isolates from Burundi. The nested PCR products of seven randomly selected isolates were purified and then sequenced. As the result, three allelic forms (340 bp, 370 bp, and 400 bp) were seen at the C-terminal domain of kahrp gene. The existence of multiple alleles of the kahrp gene revealed the presence of different P. falciparum strains in Burundi. It is suggested that the results could be useful in designing and the improvement of targeted therapy agents for falciparum malaria.

LACK Gene's Immune Response Induced by Cocktail DNA Vaccine with IL-12 Gene Against Cutaneous Leishmaniasis in BALB/c Mice.

BACKGROUND: Leishmaniasis is caused by parasitic protozoa of the genus Leishmania which is an obligate intracellular parasite in the infected host. Individuals who have been recovered from clinical leishmaniasis develop strong immunity against reinfection. DNA vaccines are the new type of vaccines that induce expression of protein eukaryotic cells. DNA vaccines can be stimulated by the cellular and humoral immune responses using one or several genes. METHODS: A DNA vaccine containing plasmids encoding the pcLACK+pcTSA genes of Leishmania major (L. major) (MHRO/IR/75/ER) in the vicinity of IL-12 gene expression was made and then its protective efficacy in comparison with single-gene of LACK was evaluated. Also, BALB/c mice were immunized intramuscularly three times. The humoral and cellular immune responses were evaluated after immunization with pcLACK, pcLACK+pcTSA+pCAGGS-IL12, and then challenged with L. major. RESULTS: Humoral response and IFN-γ values were significantly higher than control groups after immunization with pcLACK, pcLACK+pcTSA+pCAGGS-IL12 and challenge with L. major (p≤0.05). IL-4 values were increased in the control groups in such a way that they were remarkably higher than the pcLACK, pcLACK+pcTSA+ pCAGGS-IL12 groups (p≤0.05) after immunization and challenge with L. major. CONCLUSION: The survival time of the immunized mice with pcLACK, pcLACK+pcTSA+ pCAGGS-IL12 groups was higher than the control groups. Then, DNA vaccine of pcLACK appeared to be likely able to induce more protection against infection with L. major in mice. Therefore, cocktail DNA is effective to enhance specific immunity.

Pediatric Vulvar Bleeding due to Leech Infestation: A Case Report.

Leech infestation most frequently occurs in upper body cavities of children including pharynx, nose and esophagus and, more rarely, the vagina and vulva. Here we describe a 6-yr-old girl with vulvar bleeding caused by leech infestation that referred to the Emergency unit of the Booali Sina Hospital in Sari, Iran in Sep 2015. She had a history of swimming in a pond prior to the occurrence. The leech infestation particularly vulvar involvement among young girls is extremely rare and yet neglected event in the world.

Global status of visceral leishmanial infection among blood donors: A systematic review and meta-analysis.

INTRODUCTION: Transmission of Leishmania through transfusion has been reported from various Visceral leishmaniasis (VL) endemic areas of the world. The true burden of Leishmania infection in blood donors remains generally unknown. Thus, the present systematic review attempted to determine the global prevalence of Leishmania infection among blood donors. METHODS: Data were extracted through five English and five Persian databases during the period from 1997 to 2016. Overall, 16 articles fulfilled the inclusion criteria and were used for data extraction in this systematic review. RESULTS: In total, 13,743 blood donors from different regions of world were examined. The prevalence rate of Leishmania infection according to seropositivity obtained 7% (95%CI: 5%, 8%). The lowest and the highest prevalence were related to Bangladesh 0.25% (95%CI: 0.0%, 1.0%) and Brazil, 16% (95%CI: 12%, 19%). Seroprevalence rate of leishmaniasis among females was more (4.60%) than males. Of 15 studies included in the meta-analysis, the pooled prevalence rate of molecular tests was obtained 2% (95%CI: 1%, 3%) in which Iran and Spain had the lowest and the highest prevalence, 0.05% and 7%, respectively. Our analysis showed that L. infantum was more common than L. donovani as etiological agent of VL among all donors. CONCLUSION: Our data confirms the presence of asymptomatic carriers of VL in endemic areas and supplies as an attentive to the likelihood of these carriers acting as blood donors. Moreover, we conclude that molecular tests for screening in asymptomatic blood donor provide an accurate estimate of the rate of infection over serological tests.

Medicinal plants with promising antileishmanial activity in Iran: a systematic review and meta-analysis.

BACKGROUND: Leishmaniasis is a major public health problem worldwide. The aim of the present study was to investigate medicinal plants with anti-Leishmania activity which used in Iran. METHODS: Data were systematically gathered from five English databases including Ebsco, Science Direct, PubMed, Google Scholar and Scopus, four Persian databases including Magiran, Iran doc, Iran medex and the Scientific Information Database (SID) from 1999 to April 2015. Information obtained included plant family, extraction method, concentrations of extracts, animal models and parasite strains. RESULTS: A total of 68 articles including 188 experiments (140 in vitro and 48 in vivo) between 1999 and 2015, met our eligibility criteria. Thoroughly, 98 types of plants were examined against three genera of Leishmania spp. For the heterogeneity study conducted, it was showed that there was a great deal of variation among studies. Based on random effect, meta-analysis pooled mean of IC50 was obtained 456.64 (95% CI: 396.15, 517.12). CONCLUSION: The most Iranian plants used as anti-leishmanial activity were Artemisia species, Allium sativum, Achilleamille folium, Peganum harmala and Thymus vulgaris. The present systematic and meta-analysis review provide valuable information about natural products with anti-Leishmania activity, which would be examined in the future experimental and clinical trials and herbal combination therapy.