Cutaneous Leishmaniasis Lesion on the Ear from Kashan, Central Iran: A Case Report.

Cutaneous leishmaniasis (CL) frequently occurs in many rural and urban areas of Iran. Leishmania major and L. tropica are principally two causative agents of CL in Iran. We report here a case of leishmaniasis of the ear in a 61-year-old man referred to the Reference laboratory, Kashan, central Iran, in Jan 2022. He suffered from a 2-month history of a 1×3 cm lesion on the left ear. In the microscopy examination, amastigotes forms of Leishmania spp. were observed. L. tropica was confirmed using a single PCR with species-specific primers. The patient was introduced to a physician to begin the treatment protocol. It is recommended that physicians, especially in an endemic area, investigate any atypical lesion for CL.

Patterns of cutaneous leishmaniasis during the COVID-19 pandemic in four endemic regions of Iran.

BACKGROUND: Both zoonotic cutaneous leishmaniasis (ZCL) and anthroponotic cutaneous leishmaniasis are endemic in different regions of Iran. It is important to know the type of cutaneous leishmaniasis (CL) for prevention strategies. On the other hand, Iran is one of the countries that was severely affected by the coronavirus 2019 (COVID-19) pandemic. However, little is known about the causative agents of CL during the COVID-19 pandemic in Iran. METHODS: In this study, a total of 181 samples were isolated from patients with CL lesions in four different endemic cities of Iran (Jahrom, Juyom, Kashan and Shoushtar) during the COVID-19 pandemic (the summer and autumn of 2021). The nested-PCR targeting kinetoplast DNA (kDNA) minicircles was applied for species identification of the parasite and the internal transcribed spacer-1 (ITS-1) gene was applied for sequencing and phylogenetic analysis. RESULTS: Molecular detection of kDNA minicircles revealed that all isolates from the four cities were Leishmania major, indicating the ZCL pattern. Ten isolates were sequenced by the ITS-1 gene and deposited in GenBank [accession numbers: OL627363-72]. Sequencing and phylogenetic analysis displayed a high similarity rate of the isolates with sequences from other parts of Iran and Iraq. Over half of the patients (53.59%) had a single lesion, while 17.12, 14.92, 2.21 and 12.16% of cases had two, three, four and more than four lesions, respectively. About half of the patients (50.27%) had lesions on their hands, while the rest had lesions on multiple locations (19.34%), legs (16.58%), face (9.94%), ears (1.66%) and waist (2.21%). CONCLUSIONS: This study revealed the occurrence of ZCL during the COVID-19 pandemic in four endemic regions of Iran. Strategies for prevention and control of the disease should be considered to mitigate the occurrence of ZCL.

Molecular detection and genotype identification of Acanthamoeba species from bronchoalveolar lavage of patients with pulmonary symptoms suspected of cancer.

Acanthamoeba spp. are the most common free-living amoeba worldwide, inducing life-threatening diseases such as Granulomatous Amoebic Encephalitis, pulmonary infection, and amoebic keratitis. This study aimed to identify the FLA and Acanthamoeba genotypes in patients with pulmonary symptoms suspected of cancer in Kashan's hospitals, Kashan, Iran. This cross-sectional study was conducted on 97 bronchoalveolar lavage samples of patients with respiratory symptoms suspected of lung cancer, who were admitted to the Shahid Beheshti Hospital of Kashan from 2019 to 2020. The samples were cultured onto 1.5% non-nutrient agar enriched with killed Escherichia coli and examined for the presence of FLA. Following amoeba isolation and DNA extraction, Acanthamoeba spp. were determined by Polymerase Chain Reaction using JDP1 and JDP2 primers, which amplified a 490 bp fragment from the 18 S rDNA gene. Eighteen Acanthamoeba isolates were sequenced, and the genotypes were identified. The prevalence of FLA and Acanthamoeba and the relationship between symptoms and demographic variables were analyzed with SPSS Software version 16. The prevalence rates of FLA and Acanthamoeba in the BAL samples was 86.6% and 73.2%, respectively. All Acanthamoeba isolates belonged to the T4 genotype. The most symptoms among Acanthamoeba-positive patients were dyspnea and cough; however, their difference was not statistically significant. The findings indicated the high prevalence of FLA and Acanthamoeba in BAL in the population suspected of cancer in Kashan. Since the T4 genotype is a pathogenic genotype of Acanthamoeba, training health and improving sanitation levels would help to prevent infection.

Disseminated scabies in a 2-month-old infant.

Scabies is a skin disease caused by the mite Sarcoptes scabiei. We report disseminated scabies in a 2-month-old girl as well as eczematoid lesions in her mother. The diagnosis was made by skin scraping and microscopic examination of the crusts. The patients were successfully treated with permethrin cream (5%).

Leishmaniasis and Trace Element Alterations: a Systematic Review.

Leishmaniasis is a worldwide prevalent parasitic infection caused by different species of the genus Leishmania. Clinically, the disease divided into three main forms, including visceral leishmaniasis (VL), cutaneous leishmaniasis (CL), and mucocutaneous leishmaniasis (MCL). There is no vaccine for human leishmaniasis and their treatment is challenging. Trace elements (TEs) alteration, including the selenium (Se), zinc (Zn), copper (Cu), ron (Fe), and magnesium (Mg) have been detected in patients with CL and VL as well as canine leishmaniasis. Because TEs play a pivotal role in the immune system, and host immune responses have crucial roles in defense against leishmaniasis, this systematic review aimed to summarize data regarding TEs alteration in human and animal leishmaniasis as well as the role of these elements as an adjuvant for treatment of leishmaniasis. In a setting of systematic review, we found 29 eligible articles (any date until October 1, 2020) regarding TEs in human CL (N = 12), human VL (N = 4), canine leishmaniasis (N = 3), and treatment of leishmaniasis based on TEs (N = 11), which one study examined the TEs level both in CL and VL patients. Our analysis demonstrated a significantly decreased level of Fe, Zn, and Se among human CL and canine leishmaniasis, and Zn and Fe in patients with VL. In contrast, an increased level of Cu in CL patients and Cu and Mg in VL patients and canine leishmaniasis was observed. Treatment of CL based zinc supplementation revealed enhancement of wound healing and diminished scar formation in human and experimentally infected animals. The results of this systematic review indicate that the TEs have important roles in leishmaniasis, which could be assessed as a prognosis factor in this disease. It is suggested that TEs could be prescribed as an adjuvant for the treatment of CL and VL patients.

Scolicidal Effects of Chitosan-Curcumin Nanoparticles on the Hydatid Cyst Protoscolices.

PURPOSE: In the current era, cystic echinococcosis (CE), as larval stage of Echinococcus granulosus, is considered as a threat to human health. Scolicidal agents used in the surgery of cysts have different side effects. Therefore, the present study aimed to assess the effects of chitosan nanoparticles containing curcumin (Ch-Cu NPs) on the protoscolices of the hydatid cyst in vitro. METHODS: Ch-Cu NPs were synthesized using a simple co-precipitation method and their structural and morphological properties were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), zeta analyzer, and Fourier transform infrared (FT-IR) spectroscopy. Then, the effects of different concentrations of Ch-Cu NPs (0.25, 0.05, 1, 2, and 4 mg/mL) on the fatality rate, and the length and width of protoscolices in different times (5, 10, 20, 30, and 60 min) were investigated. In addition, the SEM technique was used to evaluate the structure of the protoscolices after treatment. RESULTS: Based on the results, the presence of curcumin on the chitosan nanoparticles was confirmed by FT-IR analysis. Further, XRD analysis approved the crystal structure of chitosan NPs. Furthermore, the highest fatality rate was 68% in 4 mg/mL concentration of Ch-Cu NPs. The length and width of protoscolices decreased based on the high concentrations of Ch-Cu NPs, compared to the control group. CONCLUSION: Finally, Ch-Cu NPs expressed good scolicidal activities, which made them suitable to be considered as an anti-protoscolex agent.

Genetic Variation of Giardia lamblia Isolates from Food-handlers in Kashan, Central Iran.

BACKGROUND: Based on genotyping study of human isolates of Giardia lamblia; humans are mainly infected by two assemblages A and B. The present study was carried out to determine the sub-assemblages of G. lamblia isolated from food handlers referred to Kashan health centers, central Iran, 2015. METHODS: In this cross-sectional study, 3653 stool samples collected from food-handlers that annually refer to health center for getting a health certification and examined microscopically for G. lamblia cyst. Totally, 44 isolates were selected from 47 Giardia positive samples. Cysts were partially purified by the sucrose density gradient method. After freeze-thaw cycles, genomic DNA was extracted using QIAamp Stool Mini kit. A single step PCR-RFLP method was used to amplify a 458bp fragment at the glutamate dehydrogenase (gdh) locus, restriction enzymes BspLI and RsaI were used for distinguish between genotypes A and B and their subgroups. RESULTS: Of 44 isolates, 24(54.5%) were sub-assemblage AII, 9(20.5%) group B including 7(15.9%) BIII and 2(4.6%) BIV sub-assemblage and 11(25%) isolates showed a mixed pattern of AII and B. Sub-assemblage AI was not detected in this study. CONCLUSION: The higher rate of sub-assemblage AII demonstrated an anthroponotic origin of the infection so infected food-handlers could directly transmit this protozoan to consumers via contaminated food and water. For finding of pattern of transmission and distribution of Giardia assemblages and sub-assemblage, more studies in human and animal population in different regions are necessary.

Intestinal parasitic infections in different groups of immunocompromised patients in Kashan and Qom cities, central Iran.

INTRODUCTION: Intestinal parasitic infections (IPIs) are important causes of morbidity and mortality in patients with immunocompromising conditions. OBJECTIVE: The aim of this study was to determine the prevalence of IPIs in different groups of immunocompromised patients, including hemodialysis patients (HD), renal transplant recipients (RTR), cancer and HIV/AIDS patients in comparison with healthy individuals in two central cities of Iran (Kashan and Qom). METHODS: In this case-control study, the stool samples of 135 HD, 50 RTR, 60 cancer patients, 20 HIV/AIDS patients and 120 healthy subjects were tested using direct-smear, formol-ether concentration, Ziehl-Neelsen staining and Agar plate method. RESULTS: The overall infection rate was 11.7% (31/265) in patient groups and 0% (0/120) in the control group. The frequency of parasites was 25% in HIV/AIDS patients, 11.9% (16/135) in HD, 12.0% (6/50) in RTR and 6.7% (4/60) in cancer patients. Blastocystis hominis (4.2%) and Giardia lamblia (3.0%) were the most prevalent parasites in patient groups. The infection rate was significantly higher in male (17.6%) than female (5.4%) patients (p = .002), but no statistically significant association was observed according to the age and educational levels. CONCLUSIONS: This study showed a high prevalence of IPIs in immunocompromised patients. The results of this study suggest that periodic stool examinations for screening of IPIs should be included as a part of routine medical care in these patients.

Serological and molecular survey of toxoplasmosis in renal transplant recipients and hemodialysis patients in Kashan and Qom regions, central Iran.

Toxoplasma gondii is one of the important opportunistic pathogen among solid-organ transplant recipients and hemodialysis patients (HD). This study was aimed to detect toxoplasmosis among 50 renal transplant recipients (RTR), 135 HD and 120 healthy individuals in two cities (Kashan and Qom) that located in the center of Iran, from 2014 to 2015. Serological detection (IgG and IgM antibodies) was performed among all individuals in case and control groups. Molecular detection was performed on all IgM positive individuals or IgG positive with moderate to high (>51 IU/mL) antibody titers in HD (n = 42) and control groups (n = 21). In RTR patients, molecular detection was conducted among all seropositive or seronegative individuals (n = 50). IgG seropositivity was detected in 52% (26/50) of RTR, 63% (85/135) of HD and 33.3% (40/120) of the control group. The rate of anti-T. gondii IgG antibody was significantly elevated in RTR and HD patients than the control group (p = 0.023 and p < 0.001, respectively). IgM seropositivity was only detected in one HD patient. T. gondii DNA was detected in 12% (6/50) of RTR and 7.1% (3/42) of HD patients. The results of this study suggested that the screening of toxoplasmosis should be given greater consideration among RTR and hemodialysis patients.

Comparison of Molecular, Microscopic, and Culture Methods for Diagnosis of Cutaneous Leishmaniasis.

Cutaneous leishmaniasis (CL) is endemic in the northwest of Isfahan province, Iran. Increase in the incidence of the disease in Kashan has made it necessary to find out the best method for diagnosis and molecular characterization of Leishmania species. In the present study, 130 patients suspected to cutaneous leishmaniosis referred to health care centers of Kashan were examined. Serosity of lesion was collected for smear preparation and cultured in Novy-Nicolle-McNeal medium. DNA was extracted from serosity, and Leishmania species was determined by polymerase chain reaction (PCR) and nested PCR using kinetoplast DNA (kDNA) specific primers. The diagnostic criteria of CL were based on the observation of amastigotes in the smear, promastigotes in culture, presence of expected bands in PCR, or nested PCR. Of 130 specimens, 87 (66.9%), 72 (56.2%), 98 (75.4 %), 96 (73.8%), and 99 (76.2%) were positive for microscopic culture, PCR, nested PCR, and combined PCR and microscopy (proposed method), respectively. Sensitivity, specificity, positive and negative predictive values of PCR were 99%, 100%, 100%, 96.9%, respectively, for microscopy 87.9%, 100%, 100%, 72.1%, for culture 72.7%, 100%, 100%, 53.4 %, and for nested PCR 97%, 100%, 100%, 91.2%, respectively. Based on the results of the study, kDNA-PCR was the most sensitive method for diagnosis of CL.

Uncommon Human Urinary Tract Myiasis Due to Psychoda Sp. Larvae, Kashan, Iran: A Case Report.

Contamination of human and animal body tissues with flies' larvae and diptera cause myiasis. A 26 yr old female patient refers to Kashan Shahid Beheshti Hospital, central Iran because of urogenital infection, pain in the right part of stomach, smelly and reddish vaginal discharge and frequent urination. In the first checking, urine sample was taken. In the sample, active and alive larvae were seen. The live samples were taken to the Environmental Health Department Lab of Kashan University of Medical Sciences in clean glass jars. In the morphological survey, Psychoda sp larvae were identified. In Iran, this study is the first report of this species of larva that causes urinary myiasis. This fly larva is not carnivore or bloodsucker and feeds on bacterial agents. Observance of personal hygiene especially during defecation and urination is essential to prevent contamination of this type of myiasis.

ToRCH "co-infections" are associated with increased risk of abortion in pregnant women.

ToRCH infections (toxoplasmosis, rubella, cytomegalovirus and Herpes simplex virus) have long been known to be associated with bad obstetric outcomes. However, little information is available about the impact of ToRCH co-infections on the outcome of pregnancy. Hence, we tested the IgG and IgM antibodies to Toxoplasma gondii, Rubella, Cytomegalovirus and Herpes Simplex Virus among 81 pregnant women with abortion (case group) and 98 pregnant women with normal delivery (control group). In the single-infection model, only CMV-IgM seropositivity was significantly increased in case than control group (25.9% in case and 12.2 % in control, OR = 2.5, P = 0.019). In the co-infection model, 14 patterns were recognized, but two patterns were significantly increased in the case than the control group. Co-infection of T. gondii IgG + CMV IgM was 9.1-fold increased in the case than the control group (8.6% in the case and 1% in control, OR = 9.1; P = 0.024). Also, co-infection of T. gondii IgG + HSV IgG + CMV IgM was 7.7-fold increased in case than the control group (7.4% in case and 1 % in control, OR = 7.7; P = 0.04). Although the OR of other co-infections was higher in the case than the control group, the difference was not statistically significant. These findings indicate that ToRCH co-infections are associated with increased risk of abortion than single infection. Hence, the rates of co-infections should be considered in prenatal screening of ToRCH infections.

Toxoplasmosis-associated abortion and stillbirth in Tehran, Iran.

OBJECTIVES: This study was aimed to evaluate the role of toxoplasmosis in etiology of abortion and stillbirth based on molecular and serological techniques. MATERIAL AND METHODS: A total of 110 pregnant women with abortion and stillbirth were enrolled as the case group, and 110 pregnant women with normal delivery were enrolled as the control group. Serological and molecular detections of Toxoplasma gondii were assessed by ELISA and PCR methods. RESULTS: The seroprevalence of IgG was 25.5% in the case group (26.8% in abortion and 21.4% in stillbirth) and 26.4% in the control group. IgM seropositivity was detected in 2.7% of the case group (3.6% in abortion and 0% in stillbirth) and 0.9% of the control group (p = 0.37). Toxoplasma gondii DNA was detected in 6.4% of the case group (7.3% in abortion and 3.6% in stillbirth) and 1.8% of the control group by PCR (p = 0.17). The major risk factor of congenital toxoplasmosis was the history of eating undercooked meat (p = 0.06). CONCLUSION: Results of this study revealed that the rate of PCR positive in women with abortion and stillbirth was 3.7 times higher than that in normal delivery, but the difference was not statistically significant. These findings suggest that toxoplasmosis can be involved in etiology of abortion and stillbirth.

Molecular Diagnosis of Clinical Isolates of Cutaneous Leishmaniasis Using ITS1 and KDNA Genes and Genetic Polymorphism of Leishmania in Kashan, Iran.

Cutaneous leishmaniasis is a common skin disease caused by leishmania parasite. An accurate diagnosis of parasites species is possible using molecular techniques. This study was carried out to compare internal transcribed spacer (ITS1) and kinetoplast deoxyribonucleic acid (KDNA) genes for identifying Leishmania species by Polymerase Chain Reaction (PCR), furthermore, genetic diversity of isolates was studied. This research examined 130 patients who were suspected of cutaneous leishmaniasis and referred to Kashan's health centers from 2011-2014. After DNA extraction from serosity, PCR were performed using ITS1 and KDNA primers. Cutaneous Leishmaniasis was diagnosed by the observation of 320 bp band in the ITS1-PCR. The PCR products were digested with restriction enzyme HaeIII and then leishmania species were identified by patterns of enzymatic digestion. The diagnostic criteria of Cutaneous Leishmaniasis (CL) in KDNA-PCR were based on the observation of 760 and 650 bp for Leishmaniasis tropica and Leishmaniasis major, respectively. Twelve isolates of leishmania were sequenced and the phylogenetic tree was traced using the results of sequencing by Mega 4 software. Out of 130 suspected patients to CL, 70 (53.8%) and 98 (75.4%) isolates were positive by Restriction Fragment Length Polymorphism (RFLP) of ITS1 and KDNA, respectively. Using ITS1 PCR, 60 samples (85.7%) and 10 samples (14.3%) were identified as L. tropica and L. major, respectively, ITS1-PCR had 25.3% false negative, compare to microscopy. While, microscopy had false negative in 13 cases compare to KDNA-PCR. Due to the lower sensitivity of the PCR-RFLP of ITS1, KDNA-PCR is recommended for diagnosis of CL. The L. tropica and L. major are the causative agents of CL.

Frequency of enteric protozoan parasites among patients with gastrointestinal complaints in medical centers of Zahedan, Iran.

We investigated the prevalence of intestinal protozoan parasites in patients with gastrointestinal complaints in medical centers in Zahedan, Iran. A total of 1562 stool samples was examined from July 2004 to January 2006 using microscopy (direct smear, formalin-ether concentration), xenic culture and PCR techniques. Four hundred and twenty-seven (27.3%) of the patients were infected with one or more intestinal parasites. Giardia lamblia (10.1%), Entamoeba coli (10%), E. hartmanni (1.7%), Blastocystis hominis (2.2%), Chilomastix mesnili (1.7%), Trichomonas hominis (0.7%), E. histolytica/E. dispar (0.51%) and Iodamoeba butschlii (0.45%) were the most prevalent protozoa detected with microscopy. Of the eight microscopy-positive E. histolytica/E. dispar samples, six were identified as E. dispar by PCR/gel electrophoresis, whereas E. histolytica was not detected at all. Although Zahedan is an area with poor hygiene located in a tropical area near the border of Pakistan and Afghanistan, the prevalence of E. histolytica and E. dispar here compared with other parasites and infectious diseases is unexpectedly low.