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.

The effects of Leishmania RNA virus 2 (LRV2) on the virulence factors of L. major and pro-inflammatory biomarkers: an in vitro study on human monocyte cell line (THP-1).

BACKGROUND: Cutaneous Leishmaniasis (CL) is a parasitic disease with diverse outcomes. Clinical diversity is influenced by various factors such as Leishmania species and host genetic background. The role of Leishmania RNA virus (LRV), as an endosymbiont, is suggested to not only affect the pathogenesis of Leishmania, but also impact host immune responses. This study aimed to investigate the influence of LRV2 on the expression of a number of virulence factors (VFs) of Leishmania and pro-inflammatory biomarkers. MATERIALS AND METHODS: Sample were obtained from CL patients from Golestan province. Leishmania species were identified by PCR (LIN 4, 17), and the presence of LRV2 was checked using the semi-nested PCR (RdRp gene). Human monocyte cell line (THP-1) was treated with three isolates of L. major with LRV2 and one isolate of L. major without LRV2. The treatments with four isolates were administered for the time points: zero, 12, 24, 36, and 48 h after co-infection. The expression levels of Leishmania VFs genes including GP63, HSP83, and MPI, as well as pro-inflammatory biomarkers genes including NLRP3, IL18, and IL1ß, were measured using quantitative real-time PCR. RESULTS: The expression of GP63, HSP83, and MPI revealed up-regulation in LRV2 + isolates compared to LRV2- isolates. The expression of the pro-inflammatory biomarkers including NLRP3, IL1ß, and IL18 genes in LRV2- were higher than LRV2 + isolates. CONCLUSION: This finding suggests that LRV2 + may have a probable effect on the Leishmania VFs and pro-inflammatory biomarkers in the human macrophage model.

Isolation and genotyping of Acanthamoeba species and Vahlkampfiidae in the harsh environmental conditions in the centre of Iran.

Different species of free-living amoeba (FLA) have been abundantly isolated in harsh environmental conditions such as hot springs and brackish water. The present study aimed to isolate, genotype, and evaluate the pathogenicity of FLAs in Qom Roud, a large river, in the centre of Iran. About 500 mL of water samples (n = 30) were collected from each sampling site and were investigated for the presence of FLAs using morphological and molecular characters. Genotype identification was performed using DNA sequencing and a phylogenetic tree was constructed with the MEGA X software. The pathogenic potential of all positive isolates was evaluated using the tolerance ability test. Morphological and molecular analysis indicated that 14 (46.66%) and two (6.66%) water samples were positive for Acanthamoeba species and Vahlkampfiidae, respectively. According to sequence analysis, Acanthamoeba isolates related to the T4 genotype and Vahlkampfiidae sequences were similar to Naegleria philippinensis. In the next step, thermo- and osmotolerance tests indicated four Acanthamoeba strains are extremely pathogenic. Our data showed the presence of potentially pathogenic Acanthamoeba T4 genotype and N. philippinensis in the super harsh Qom Roud. Contamination of water with virulent T4 genotype of Acanthamoeba may pose risk factors for contact lens users, children, and immunocompromised people.

Development of an Indirect Fluorescent Antibody (IFA) Assay for the Detection of Leishmania RNA Virus 2 (LRV2) in Leishmania Parasites.

Background: 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. Methods: 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. Results: 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. Conclusion: 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.

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 identification and genotyping of Acanthamoeba spp., in bronchoalveolar lavage fluid from immunocompetent patients with chronic respiratory disorders (CRD).

This study aimed to investigate the presence and genotyping of Acanthamoeba spp., in the bronchoalveolar lavage fluid (BALF) of immunocompetent patients with chronic respiratory disorders (CRD). In this study, 211 BALF samples were collected from patients with CRD during the COVID-19 pandemic who were candidates for fiberoptic bronchoscopy (FOB) at Imam Khomeini Hospital, Sari, Mazandaran Province, northern Iran and investigated for Acanthamoeba spp., by PCR. A total of 211 FBAL samples were examined; 5 (5/211; 2.36%) were positive by using the PCR test for Acanthamoeba spp. According to sequence analysis, three strains belonged to the T4 genotype and one strain to the T2 genotype. Our data demonstrate that the presence of Acanthamoeba (T4 and T2) in BALF specimens of patients with respiratory infections. However, it is important to note that these findings may be merely accidental. Our findings suggest further investigation to fully understand the role of Acanthamoeba spp. in the pathogenesis of lung infections.

Leishmania RNA virus 2 (LRV2) exacerbates dermal lesions caused by Leishmania major and comparatively unresponsive to meglumine antimoniate treatment.

PURPOSE: The present study investigated the possible role of Leishmania RNA virus 2 (LRV2) in the severity of dermal lesions and treatment failure due to Leishmania major. METHODS: The drug susceptibility of 14 clinical isolates of L.major, including resistant (n = 7) and sensitive (n = 7) isolates, was checked in the J774A.1 macrophage cell line. The presence of LRV2 among isolates was investigated by the RdRp gene and semi-nested PCR. Moreover, 1 × 106 sensitive L. major LRV2+ and LRV2- promastigotes were inoculated subcutaneously into the base tails of the 40 BALB/c mice divided into 4 groups (n = 10 in each group), including clinical LRV2+, clinical LRV2-, positive control LRV2+ and negative control LRV2-. The groups were infected with a unique isolate. The lesion size and parasite burden were evaluated. RESULTS: Sensitive and resistant isolates were determined by the drug susceptibility method. A higher presence of LRV2 was observed among MA-resistant isolates (6/7) compared with susceptible isolates (4/7), which was not statistically significant (P = 0.237). On the other hand, a comparison of the lesion sizes between the LRV2+ and LRV2- BALB/c mice groups revealed that the mean size of the lesion in the LRV2+ groups was significantly higher than the LRV2- (P = 0.034). In the same direction, there was an increased parasite burden in mice inoculated with LRV2+ groups compared with the LRV2- BALB/c mice groups (P = 0.002). CONCLUSIONS: Our findings showed that the presence of LRV2 could be one of the factors contributing to exacerbating CL. Although we found a higher presence of LRV2 in the resistant isolates, it seems that further investigations are recommended to determine the detailed association between lesions' aggravation and being comparatively unresponsive to treatment.

Successful Isolation of Leishmania RNA Virus (LRV) from Leishmania major in a Cutaneous Leishmaniasis Focus in Central Iran: An Update on Cases.

PURPOSE: Cutaneous leishmaniasis (CL) is a major vector-borne disease that affects people globally, including Iran. Different factors are associated with leishmaniasis pathogenicity; recently, a link of the possible relationship between Leishmania RNA Virus (LRV) and disease severity was proposed, especially in the New World leishmaniasis (NWL). This study was aimed to investigate the presence of LRV2 in Leishmania isolates in Aran o Bidgol, Isfahan province. METHODS: Samples were collected from 110 CL-suspected patients referred to the health center. In this study, we aimed to investigate CL cases (parasitologically and clinically), identify Leishmania species (by ITS1-PCR-RFLP), and finally detection of LRV2 (by RdRp-semi-nested PCR). RESULTS: Parasitological methods showed 60 positive cases, based on the HaeIII enzyme restriction profile, 59 cases were caused by L. major and 1 case by L. tropica. Our project is the first study on LRV2 isolation in Aran o Bidgol city and the LRV was successfully detected from a single L. major isolated in a women's hand lesion. Using BLAST, 94.8-100% similarity was observed in the RdRp sequence of current LRV isolate with those available in GenBank from Iran or overseas. CONCLUSION: L. major was the main cause of CL in Aran o Bidgol, although L. tropica is also present in a much lower proportion in the area. This is the first report on the presence of LRV2 in Aran o Bidgol and the fifth in Iran.

First Co-morbidity of Lophomonas blattarum and COVID-19 Infections: Confirmed Using Molecular Approach.

INTRODUCTION: Lophomoniasis is caused by Lophomonas spp., a new emerging protozoan, which commonly affects the human lower respiratory tract. The Lophomonas parasite mostly lives commensally in the hindgut of cockroaches. CASE PRESENTATION: We present the case of a 33-year-old woman, 30 weeks pregnant, who had severe COVID-19. She was intubated upon admission and began the routine COVID-19 treatment. To rule out possible super infection dual with COVID-19, microscopic examination of the patient's mini-bronchoalveolar lavage (mini-BAL) specimen, revealed L. blattarum, which was identified by the SSU rRNA-PCR and sequencing approaches (accession number: MZ093069). According to that, the patient was treated successfully with metronidazole. CONCLUSION: To prevent serious complications, lophomoniasis should be listed in co-morbidity cases of COVID-19 infection during the COVID-19 pandemic worldwide. To the best of our knowledge, this is the first co-infection of Lophomonas blattarum and COVID-19 in the world which has been confirmed using a molecular approach.

In vivo anti-leishmanial activity of concocted herbal topical preparation against Leishmania major (MRHO/IR/75/ER).

Cutaneous leishmaniosis (CL) is considered as one of the most important tropical diseases. Herbal therapy is the ideal treatment for CL because of the reduced injection pain, availability, lower cost and non-toxicity effects. The present study aimed to evaluate the in vivo antileishmanial activity of concocted herbal topical preparation (Aloe vera, Perovskia abrotanoides, Nigella sativa, propolis, lavender and olive oil) to evaluate its efficacy against Leishmania major (MRHO/IR/75/ER) in comparison to the gold standard treatment. Following the cause of cutaneous leishmaniosis, the BALB/c mice were divided into three groups, test group (ointment formulation), positive control (Glucantime) and negative control (untreated), respectively, which were treated twice a day for 28 consecutive days. The lesion size and parasite burden were evaluated for in vivo evaluation. The herbal topical ointment was able to significantly decline the lesion progression and reduce parasite burden in mice inoculated with L. major promastigotes in the test group compared with the negative control group (P=<0.001). In mice treated with the formulation, the number of amastigotes significantly decreased (P=<0.001), compared with that in the negative control group. Moreover, comparative features of both treatments showed there was no difference between the herbal-treated and glucantime-treat mice (P=0.63). The herbal topical ointment displayed significant in vivo antileishmanial activities. It may be that using ointment formulation beside other skin repair compounds can be used as an alternative medicine in the treatment and healing of human CL lesions. Further investigations are needed to study the pharmacologic and pharmacokinetics aspects of ointment formulation in the treatment and healing of human CL lesions.

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.

Molecular Identification of Pathogenic Free-Living Amoeba from Household Biofilm Samples in Iran: A Risk Factor for Acanthamoeba Keratitis.

Free-living amoeba (FLA) are ubiquitously distributed in the environment. However, they are also the causative agents of opportunistic infections in humans and other animals. A biofilm comprises any syntrophic consortium of microorganisms in which cells stick to each other and often also to a surface. Moreover, FLA have been detected in various biofilms around the world. Therefore, the present study aimed to check for presence of FLA in samples from household biofilms in Iran and to characterize them at the molecular level. A total of 69 biofilm samples collected from showerheads, kitchen areas, and bathroom sinks were analyzed. Positive samples for FLA were characterized at the morphological and molecular levels. Furthermore, the results of morphology analysis indicated that 26.08% (18/69) of biofilm samples were positive for Acanthamoeba spp., Vermamoeba genus, and Vahlkampfiids. According to sequence analysis, five strains of Acanthamoeba isolates related to the T4 genotype and two strains belonged to the T2 genotype. In addition, the pathogenic potential of Acanthamoeba-positive isolates was conducted using the tolerance ability test. The results of BLASTn of Vermamoeba sequences were similar to what was expected for Vermamoeba vermiformis. The above-mentioned reasons revealed that the relative high contamination of household biofilm samples with FLA may pose a risk for people using soft contact lenses and for patients with traumatic cataract. Our finding proposes that filtration should be performed in shower heads and indicates the need to monitor people at increased risk of Acanthamoeba keratitis.

The Geographical Distribution of Human Cutaneous and Visceral Leishmania Species Identified by Molecular Methods in Iran: A Systematic Review With Meta-Analysis.

Leishmaniasis is one of the most common vector-borne parasitic diseases in Iran. Leishmania species identification is necessary for epidemiological aspects, precise prognosis, control and treatment of the disease. We systematically searched all the studies, reports, and documentation related to species identification and geographical distribution of causative agents of cutaneous (CL), mucosal (ML), and visceral leishmaniasis (VL) using DNA-based molecular diagnostic techniques in Iran. International databases including PubMed, ScienceDirect, Embase, Google Scholar, Scopus, and Web of Science were systemically searched for English articles and Iran's databases including SID, IranMedex and Magiran were searched for Persian reports and articles. Searches were performed from 1999 to 2019 (20 years). The current review was conducted using the keywords: cutaneous leishmaniasis, visceral leishmaniasis, Leishmania species, Human, Molecular, PCR, and Iran. The study quality was evaluated using the NOS checklist. This meta-analysis procedure was accomplished using STATA, version 2.7.9. Of the 3,426 records identified in the initial search, 154 articles met inclusion criteria and qualified for the systematic review and meta-analysis. In subgroup analysis, the pooled frequency of causative agents of CL isolates was 67.3% (95% CI: 59.51-74.67%) for L. major and 32.1% (95% CI: 24.72-39.87%) for L. tropica. In addition, the pooled frequency of causative agents of VL isolates was 97.1% (95% CI: 94.6-98.8%) for L. infantum and 2.9% (95% CI: 1.12-5.37%) for L. tropica. The findings of this study showed that the main causative agents of CL and VL in Iran are L. major and L. infantum, respectively. Moreover, kinetoplast DNA (kDNA) and internal transcriber spacer (ITS) were the most used markers for identifying Leishmania species. The current study provides valuable data to encourage and direct researchers as well as public health managers in the comprehensive leishmaniasis control and prevention planning in Iran.

Identification of Trichomonas Vaginalis Genotypes Using by Actin Gene and Molecular Based Methods in Southwest of Iran.

BACKGROUND: Trichomonas vaginalis (T. vaginalis) is a sexually transmitted protozoan parasite and the causative agent of trichomoniasis. The genetic characterization of T. vaginalis isolates shows notable genetic variation in this parasite. In the present study, we aimed to identify the T. vaginalis genotypes based on analyzing of actin gene in women specimens referred to health centers of Ilam city, southwest Iran. METHODS: A total of 1765 female samples were collected from gynecology clinics in the city of Ilam. DNA was extracted from positive samples and nested polymerase chain reaction (Nested PCR) was used to amplify the actin gene. Then, partial sequencing and genotyping of the actin gene was performed. A phylogenetic tree was drawn using the detected genotypes of T. vaginalis and reference sequences. RESULTS: Twenty-one of the 1765 urine and vaginal samples were positive for T. vaginalis. All infected individuals were married and their age in years was between 25 to 34. Further, the majority of infected women had cervical lesions, patchy erythema, and white color discharge. According to sequencing analysis, the isolates were identified as genotype G (n= 8) and genotype E (n= 2). CONCLUSION: From the collected samples, we were able to distinguish at least two genotypes (G and E) of T. vaginalis. However, lesser is known about these genotypes in the city of Ilam. Further studies with a higher number of isolates should be performed in order to understand the implications of these results in this region.

Morphological and Molecular Identification of Emerged Lophomonas blattarum Infection in Mazandaran Province, Northern Iran: First Registry-Based Study.

BACKGROUND: In the last decade, several cases of bronchopulmonary lophomoniasis (BPL) have been recorded. Little information is available about epidemiological aspects on Lophomonas infection among BPL patients. The present study was aimed to investigate the prevalence of Lophomonas spp. infection in patients who were referred to the Iranian National Registry Center for Lophomoniasis (INRCL), using morphological and molecular tests. SUBJECTS AND METHODS: We examined patients enrolled in the INRCL from 2017 to 2019 at the Mazandaran University of Medical Sciences, northern Iran. All bronchoalveolar lavage fluid (BALF) and two nasal discharges of the patients were examined by both microscopic and small-subunit ribosomal RNA (SSU rRNA) PCR methods. To confirm the species of Lophomonas, two positive samples were sequenced. RESULTS: In this study, 321 specimens (including 319 BALF and 2 nasal discharges) were microscopically examined. Lophomonas spp. was found in 45(14%) (n = 44 BAL; n = 1 nasal discharge). The mean age of infected patients was 54.9 ± 17.1 years. The following morphological characteristics were observed in both fresh and Papanicolaou-stained smears to identify Lophomonas spp. All microscopically positive specimens were confirmed with genus-specific PCR technique. The obtained sequences were deposited in Gen Bank under the accession numbers (MN243135-36). The BLAST analysis of our two sequences with the only available sequence in the Gen Bank of the Thailand strain of L. blattarum, showed identity of 99-100% and 98.51%, respectively. CONCLUSION: To the best of our knowledge, this is the first registry-based study regarding lophomoniasis worldwide. According to our study, the conventional PCR test is an available and reliable tool for confirming the Lophomonas parasite in clinical samples. Moreover, the results confirmed that L. blattarum is circulating at least in our region.

Intestinal parasites among intellectually disabled individuals in Iran: a systematic review and meta-analysis.

BACKGROUND: Poor self-care skills and personal hygiene resulted from limitations in learning and understanding, put intellectually disabled individuals at greater risk for intestinal parasitic infections (IPIs). Despite several regional reports in Iran, the overall burden on IPIs among intellectually disabled individuals is poorly understood. Hence, the present study aimed to estimate the pooled prevalence of IPIs among intellectually disabled individuals in Iran. METHODS: Using the PRISMA guidelines, we conducted a systematic review and meta-analysis of data retrieved from seven electronic databases (PubMed, Web of Science, Scopus, Embase, and ProQuest for English articles, as well as SID and Magiran for Persian) from their inception up to December 2020. Pooled prevalence was estimated using a random-effects model with a 95% confidence interval (CI) and depicted as a forest plot, while heterogeneity was evaluated using Cochran's Q-test. RESULTS: Exactly 1263 of the 3004 intellectually disabled individuals examined by 14 studies across 10 provinces of Iran were positive for IPIs. Overall pooled prevalence estimate was 41% (95% CI 29-53%) with a range of 21% (95% CI 10-32%) to 68% (95% CI 55-80%) across sub-groups. Entamoeba coli (16.2%; 95% CI 10.3-22%), Blastocystis spp. (12.2%; 95% CI 7.2-17.2%), and Giardia duodenalis (11.9%; 95% CI 7.4-16.3%) were the most prevalent protozoan species. In terms of helminthic agents, the most prevalent species were Enterobius vermicularis (11.3%; 95% CI 6.3-16.3%) followed by Strongyloides stercoralis (10.9%; 95% CI 5.0-16.9%) and Hymenolepis nana (2.8%; 95% CI 0.4-5.2%) CONCLUSION: IPIs are highly prevalent among intellectually disabled individuals in Iran. Improving the health status and implementing infectious disease prevention strategies in rehabilitation centers, health promotion interventions to improve personal hygiene of intellectually disabled individuals, as well as utilize sensitive diagnostic methods besides routine stool examination techniques, and treatment of infected individuals will help in the control of these infections among intellectually disabled individuals.

Expression analysis of centrin gene in promastigote and amastigote forms of leishmania infantum iranian isolates: a promising target for live attenuated vaccine development against canine leishmaniasis.

BACKGROUND: Leishmania parasites express various essential proteins in different growth phases (logarithmic/stationary) and forms (promastigote/amastigote). Targeting the genes encoding such proteins paves the way for controlling these parasites. Centrin is an essential gene, which its protein product seems to be vital for the proliferation of Leishmania parasites. Herein, this study was contrived to analyze the expression level of the centrin gene in different growth phases and forms of Leishmania infantum (L. infantum) parasites isolated from various endemic areas of canine leishmaniasis (CanL) in Iran. RESULTS: All three collected isolates were identified as L. infantum using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Real-time reverse transcription (RT)-PCR revealed a statistically significant up-regulation (3.13-fold) in the logarithmic phase promastigotes compared to stationary ones (p < 0.01), whereas centrin was expressed equally in intracellular amastigotes at different time points during cell culture. Also, our finding revealed a slight up-regulation of the centrin gene (1.22-fold) in the intracellular amastigotes compared to logarithmic phase promastigotes, which was found statistically non-significant (p > 0.05). CONCLUSIONS: Centrin gene in Iranian isolates of L. infantum is more expressed in exponential than stationary phases and seems to be considered as a promising target in the development of a genetically modified live attenuated vaccine for CanL control.

First Evidence for Colonizing of Acanthamoeba T4 Genotype in Urinary Tracts of Patients with Recurrent Urinary Tract Infections.

BACKGROUND: Limited evidence about the presence of Acanthamoeba spp. in urine specimens collected from urinary catheters of the patients in the intensive care units persuaded our study. No evidence has been found about colonizing of Acanthamoeba spp., in urinary tracts of patients with recurrent urinary tract infections (UTIs) yet. METHODS: In this study, 50 urine samples were collected from patients presenting with recurrent UTI. The type of bacteria causing UTI was determined by using bacteriological tests. To cultivate Acanthamoeba spp., in a sterile condition, 10 mL of urine was centrifuged and the sediment was cultivated on non-nutrient agar. Genotypes were determined by sequencing the DF3 region of the 18S rRNA gene. RESULTS: The bacteriological test findings on the urine samples of the UTI patients (n = 30) demonstrated that those were found to be positive for Escherichia coli (n = 17), Staphylococcus aureus (n = 6), Pseudomonas aeruginosa (n = 4) and Klebsiella spp. (n = 3) respectively. Moreover, a total of 50 urine samples was examined; 6 (6/50; 12%) and 11 (11/50; 22%) were positive by using culture and the PCR test for Acanthamoeba spp., respectively. Sequencing analysis showed all isolates were T4 genotype. CONCLUSIONS: Our data showed that the high relative prevalence of Acanthamoeba T4 genotype spp., in the urine of recurrent UTI patients. As well as, providing the first evidence for colonizing of the Acanthamoeba in the urinary tracts of patients with recurrent UTIs. These findings, warrant further investigation among those patients to fully appraise the role of Acanthamoeba spp., as possible latent carriers for resistant bacteria and biofilm formation in the future.

A Systematic Literature Review of Curcumin with Promising Antileishmanial Activity.

BACKGROUND: Curcumin (CUR) is a bright yellow chemical and it is used as an additive in foods. Recently CUR and its associated bioactive compounds have received much attention in the literature review. The aim of this systematic review is to overview the antileishmanial properties of CUR and its mechanism; perhaps the results of this study will be used for therapeutic and preventive purposes. METHODS: Following the PRISMA guidelines, international databases were systematically searched for studies published until September 2019. Articles related to the subject were selected and included in this systematic review. RESULTS: A total of 15 articles met our eligibility criteria. Then, the effect of CUR and its associated bioactive compounds on Leishmania species was evaluated. In most studies, CUR/derivatives were tested on L. major and in vitro condition. Most investigations were conducted on the promastigote rather than the more relevant intracellular amastigote stage. Our results showed that CUR overcomes the inhibitory effect of nitric oxide (NO) on Leishmania parasites. CONCLUSION: This review indicated that CUR derivatives, instead of CUR alone showed a high potential to serve as an effective herbal drug against leishmaniasis. Moreover, we concluded that the antileishmanial activity of CUR/bioactive compounds is mostly due to increased oxidative stress and apoptosis.

Toxoplasmosis among cancer patients undergoing chemotherapy: a population study based on the serological, molecular and epidemiological aspects.

BACKGROUND: Toxoplasmosis is highly prevalent in northern Iran and immunocompromised individuals are more susceptible to this infection. The present study aimed to determine the seroprevalence, parasitism and genetic diversity of Toxoplasma gondii among patients with cancer undergoing chemotherapy in northern Iran. METHODS: A total of 350 serum samples obtained from cancer patients were collected from laboratory centers in northern Iran. Immunodiagnosis and DNA detection were accomplished by ELISA and PCR. Thereafter, multiplex-nested PCR-restriction fragment length polymorphism was used for the genotyping of T. gondii. RESULTS: In general, out of 350 patients, 264 (75.4%) and 9 (2.57%) cases were positive for anti-T. gondii IgG and IgM, respectively. Moreover, 19 (5.43%) samples contained T. gondii DNA. From 19 positive samples, 10 high-quality samples with sharp and non-smear bands were selected to determine the genotypes of T. gondii. Accordingly, the samples were classified as genotype #1 (type II clonal; n=4, 40%), genotype #2 (type III clonal; n=3, 30%), genotype #10 (type I clonal; n=2, 20%) and genotype #27 (type I variant; n=1, 10%). CONCLUSIONS: As evidenced by the results, due to the high prevalence of T. gondii, cancer patients in northern Iran are at serious risk of severe toxoplasmosis and its complications. Therefore, oncologists need to regard this critical health problem as a matter requiring urgent attention.