Mini-Exon Genotyping of Leishmania Species in Khuzestan Province, Southwest Iran

Background: Leishmaniasis is a protozoan disease cause by Leishmania genus. Anthroponotic and zoonotic cutaneous leishmaniasis are endemic in Iran. The aim of this study was to identify the causative agent of cutaneous leishmaniasis by mini-exon gene in five regions of Khuzestan Province, southwest of Iran. Methods: From 2007 to 2008 in this cross-sectional study, cutaneous samples were collected from patients referred to Health Centers and Hospitals of the Khuzestan Province for cutaneous leishmaniasis diagnosis and cultured in Novy-MacNeal-Nicolle (NNN) and RPMI 1640. The propagated promastigotes were harvested and Leishmania species of cutaneous leishmaniasis were identified by RFLP and DNA sequencing of the PCR generated fragments. Results: L. major and L. tropica were the causative agents of cutaneous leishmaniasis by predominantly of L. major species. The alignment of the mini-exon sequencing isolates with reported sequencing of L. major and L. tropica revealed 92-99 identity. Conclusion: Our study showed that mini-exon PCR-RFLP was useful method to identify the causative species of cutaneous leishmaniasis

Seroprevalence of Toxoplasma gondii

Toxoplasma gondii is an obligate intracellular protozoan parasite causing toxoplasmosis in animals and humans. Primary maternal infection with toxoplasmosis during pregnancy is frequently associated with transplacental transmission to the fetus. However it is not certain whether Toxoplasma infection can cause recurrent abortion. The aim of this study was to determine the relationship between Toxoplasma infection and abortion via detection of anti-Toxoplasma gondii antibodies in sera of women with obstetrical problems and compare the results with control group consisting of women with history of normal delivery. Sera from 130 women with abortion and sera of 130 women with normal delivery were tested for IgG and IgM anti-Toxoplasma gondii antibodies by ELISA method. The present study revealed 24.6% of the samples with abortion and 21.5% of the samples with normal delivery were positive for IgG antibodies. However, statistical analysis indicated no significant differences (P>0.05). In addition, IgM antibody was detected in one woman who had aborted but not in women with normal childbirth. This study showed no significant difference between the case and control groups in IgG anti-Toxoplasma antibody but detected one sample with IgM antibodies in woman with abortion during the first trimester of pregnancy. In order to determine the relationship between Toxoplasma infection and abortion, anti-Toxoplasma IgG avidity and PCR to discriminate between recent and prior infections are recommended

Detection of Toxoplasma gondii by PCR and Mouse Bioassay in Rodents of Ahvaz District, Southwestern Iran

Toxoplasma gondii is obligate coccidian zoonotic parasite. Felidae family is definitive and wide ranges of warm-blooded vertebrates are intermediate hosts for the parasite. Rodents are measured as an important source of T. gondii infection for the definitive host. Thus, this study aimed to investigate Toxoplasm infection in rodents of Ahvaz district, southwest of Iran. A total of 100 rodents (73 Rattus norvegicus, 21 Rattus rattus, and 6 Mus musculus) were collected and studied by GRA6PCR and mouse bioassay. The finding indicated that 6 out of 100 (6%) and 2 out of 100 (2%) samples were positive by PCR and mouse bioassay, respectively. The results show notable chronic infection in the rodent and potential transmission of the infection among animal and men in the region. Accordingly, this study recommended investigating of the T. gondii infection in definitive and other intermediate hosts in other points of Khuzestan province, Southwest, Iran

The Effect of Aqueous Garlic Extract on Interleukin-12 and 10 Levels in Leishmania Major (MRHO/IR/75/ER) Infected Macrophages

Background: The aim of the present study was to investigate the immunomodulation effects of aqueous garlic extract (AGE) in the cultured macrophages infected by Leishmania major.Methods: After J774 macrophages proliferation in RPMI1640 and incubation with Leishmania for 72 hours, AGE was added in doses of 9.25, 18.5, 37, 74 and 148 mg/ml for 18, 24 and 48 hours and cell culture supernatants were harvested. The Leishmania infected J774 cells to assess the cell viability was examined using trypan blue and methylthiazol tetrazolium assay (MTT). An enzyme-linked immunosorbent assay (ELISA) was performed on cell culture supernatants for measurement of interleukin IL-10 and IL-12.Results: Dose of 37 mg/ml for 48 hours of garlic extract was the most potent dose for activation of amastigotes infected macrophages. In addition, AGE increased the level of IL-12 in Leishmania infected cell lines significantly.Conclusions: AGE treated cell is effective against parasitic pathogens, and AGE induced IL-12 differentially affected the immune response to invading Leishmania parasites

Spatial distribution and epidemiological features of cutaneous leishmaniasis in southwest of Iran

Introduction: Leishmaniasis, as a major health concern exists in 14 out of 22 countries of the Eastern Mediterranean Region (EMR). Therefore, the aim of present investigation was to evaluate the epidemiological features and spatial distribution of cutaneous leishmaniasis (CL) during six consecutive years (2009–2014). Material and methods: In current retrospective cross-sectional study among 2009–2014, simple direct smear was taken from all suspicious CL subjects who referred to health centers affiliated to Ahvaz Jundishapur University of Medical Sciences. For each patient a questionnaire including some demographic details was filled. Eventually data analysis was done by SPSS.16. Results: Trend of CL in the region was unstable. Spatial distribution of CL in central and west cities was higher than in others. During the years, a total of 4137 smear positive individuals were diagnosed. Of these 55.7% lived in urban and 44.3% lived in rural districts. Frequency of CL was higher in men (60.1%) than in women (39.9%). Also based on age range, 11–30 was the most afflicted group (45.7%). Anatomic location of ulcers was as follows: hands 45.7%, feet 27.4%, face 19.1% and other places 7.8%. Conclusions: Regarding high incidence of CL in southwest of Iran, special programs related to vector and reservoir control should be adopted and implemented. Traffic control of immigrants and travelers from neighboring endemic countries, also can be helpful

Induction of Apoptosis by Miltefosine in Iranian Strain of Leishmania infantum Promastigotes

"nBackground: Miltefosine is a new drug of choice for the treatment of visceral leishmaniasis. Numerous experi­mental studies have shown miltefosine is effective on Leishmania donovani, however, effectiveness of miltefos­ine in treatment of L. infantum is not fully understood. The aims of the present study were to evaluate cytotoxic effects of miltefosine on Iranian strain of L. infantum, and to determine its 50% inhibitory concentration (IC50) as well as lethal dose. "nMethods: Anti-L. infantum activity of miltefosine was studied by treatment of cultured promastigotes with vari­ous concentration of miltefosine. MTT assay was used to determine L. infantum viability and the results were expressed as IC50. Annexin-V FLUOS staining was performed to study apoptotic properties of this drug by us­ing FACS flow cytometry."nResults: Miltefosine led to dose-dependent death of L. infantum with features compatible with apoptosis includ­ing cell shrinkage, DNA laddering, and externalization of phosphatidylserine with preservation of integrity of plasma membrane. The 100% effect was achieved at 22 µM and IC50 after 48 hours of incubation was 7 μM."nConclusion: Miltefosine exerts cytotoxic effect on Iranian strain of L. infantum via an apoptosis-related mechanism