Cutaneous Leishmaniasis of the Eyelids: A Case Series with Molecular Identification and Literature Review

Cutaneous leishmaniasis (CL) is a protozoan disease which is endemic in Iran. It is transmitted by the Phlebotomus sand fly. The eyelid is rarely involved possibly because the movement of the lids impedes the sand fly from biting the skin in this region. Here, we report 6 rare cases of eyelid CL. The patients were diagnosed by skin scraping, culture, and PCR from the lesions. Skin scraping examination showed Leishmania spp. amastigotes in the cytoplasm of macrophages. Culture examination was positive for Leishmania spp. PCR was positive for Leishmania major and Leishmania tropica. The lesions were disguised as basal cell carcinoma, chalazion, hordeolum, and impetigo. The patients were treated with intramuscular meglumine antimoniate (20 mg/kg/day) for at least 3 weeks. They showed a dramatic response, and the lesions almost completely disappeared. We emphasized the importance of clinical and diagnostic features of lesions, characterized the phylogenetic relationship of isolated parasites, and reviewed the literature on ocular leishmaniasis.

Prevalence of Enteric Protozoan Oocysts with Special Reference to Sarcocystis cruzi among Fecal Samples of Diarrheic Immunodeficient Patients in Iran

The genus Sarcocystis is not usually considered as an important enteric pathogen in immune compromised patients. It might be expected that species for which humans are the final host (Sarcocystis hominis and Sarcocystis suihominis as well as possibly others) would be encountered increasingly often in immunodeficient persons. This study aimed to address how to detect and differentiate Sarcocystis oocysts and/or sporocysts from enteric protozoans in the diarrheal samples of immunodeficient patients in Shiraz, Iran. Diarrheal samples of 741 immunodeficient patients with recurrent persistent or chronic diarrhea were examined by microscopy and molecular biological analysis. Oocysts-positive samples were 68 Cryptosporidium spp., 9 Cystoisospora belli (syn. Isospora belli), 2 Cyclospora cayetanensis, and 15 microsporidia (Enterocytozoon bieneusi). Sarcocystis-like sporocysts found from a woman were identified as Sarcocystis cruzi through 18S rDNA amplification and phylogenetic analysis. To the best of our knowledge, this is the first report of S. cruzi from a human.

Synthesis and biological evaluation of metronidazole derivatives as anti-giardia agents

Nitroimidazole derivatives such as Metronidazole [MTZ] have been used as anti- protozoa and anti-anaerobic bacteria. In this study several compounds of MTZ derivatives were synthesized and evaluated against Giardia lamblia cyst. Firs MTZ were reacted with several alkyl halide to obtain O-alkyl MTZ derivatives, then products were purified and their chemical structures were confirmed by elemental and spectral analysis [1HNMR and Mass]. In order to biological evaluation all compounds were investigated against 25 Giardia samples isolated from giarfdiasis patients. Results showed compound 2a had the most activity on cyst of Giardia in comparison with MTZ

Recognition of pathogenic free-living amoebae in the surface water in Shiraz, Iran

Background: There are many genera of free-living amoeba in the environment, but members of only four genera [Naegleria, Acanthamoeba, Balamuthia and Sappinia] have an association with human infection. Water, soil and air are main sources of infective types of these pathogenic organisms for human

Methods: Totally, 30 samples were collected from the surface water sources of Shiraz city, the capital of Fars province, during July and August 2009. The samples were filtered and their sediments were cultured on non-nutrient agar medium and seeded with non-pathogen Escherichia coli. Then, they were incubated at three different temperatures, 22[degree]C, 37[degree]C, and 44[degree]C. The media were checked with invert microscopy and amoebae were recognized by phase-contrast microscopy and observed by light microscopy after Trichrome staining. Polymerase chain reaction [PCR] was performed for molecular detection

Results: Of the 30 samples, 29 were recognized morphologically as Acanthamoeba, the characteristics of 20 of which were confirmed by PCR. The growth rate of amoeba in 22[degree]C was more than 37[degree]C. Eight of the samples grew at 44[degree]C, but flagellate forming test and PCR were negative for Naegleria fowleri. Two of them were identified morphologically as Balamuthia and Sappinia

Conclusion: Since Fars province is located in the subtropical region where there are a lot of parks and green areas with surface water, the potential risk of diseases caused by free-living amoebae should be considered. Further investigations about various aspects of these important opportunistic protozoa are recommended especially for establishment of appropriate prevention tools

In vitro Antiplasmodial Activity and Cytotoxicity of Ten Plants Used as Traditional Medicine in Malaysia / Jurnal Sains Kesihatan Malaysia

Dichloromethane and methanolic extracts of each plant were tested for their antiplasmodial activity on chloroquineresistant strain of Plasmodium falciparum (FCB strain), based on lactate dehydrogenase activity. Cytotoxicity was assessed with the MTT test on MRC-5 human diploid embryonic lung cells. Most extracts of ten selected plants used in Malay traditional medicine in Malaysia had activity in vitro. This supports continued investigations of traditional medicine in the search for new antimalarial agent. The compounds responsible for the observed antiplasmodial effects are under investigation.

Leishmania major in Tatera indica in Estahban, Southern Iran: microscopy, culture, isoenzyme and PCR

This study identifies the infected rodent hosts to Leishmania major in Estahban town, southern Iran during 2004-2005. The rodents were caught alive from April 2004 to April 2005 in Estahban town, south of Iran and examined for any skin lesion. An impression was provided from the tissues of feet, tail, ears and any patent lesion, stained with Giemsa and studied microscopically for the presence of amastigotes. All samplings were cultured at 25°C in rabbit blood agar and considered negative if no promastigotes were visible during a two months period. The parasites from any positive culture were cryopreserved in liquid nitrogen pending their identification in PCR and isoenzyme electrophoresis. The femoral bones were histologically and ultrastrucrurally studied. Among 13 captured rodents, 8 were Tatera indica [5 male and 3 female Indian gerbils] and 5 were Rattus rattus [3 males and 2 females]. Just one female T. indica was smear-positive for amastigotes in Mohmmad Abad village. This rodent was also found culture positive for leishmanial infection which was confirmed by PCR and enzyme electrophoresis. At histological and ultrastructural levels, many clusters of amastigotes were noticed in the foamy macrophages of the femoral bone bone marrow. T. indica was found for the first time in the area and can be one of the rodents to be a potential reservoir host of L. major. It was also shown that femoral bone marrow was the tissue of choice to confirm the presence of macrophages containing the amastigote form of the parasite

leishmania infantum fml-elisa for the detection of symptomatic and asymptomatic canine visceral leishmaniasis in an endemic area of Iran

Visceral leishmaniasis [VL] is caused by Leishmania infantum in Mediterranean basin and is an endemic disease in some parts of Iran. Canines are the main reservoirs of VL in most of the endemic areas. Different serological methods have been introduced for diagnosis of canine visceral leishmaniasis [CVL]. In this survey a Fucose-Mannose Ligand [FML] ELISA, using native L. infantum antigen, was developed and its validity for detection of infected dogs in comparison with direct agglutination test [DAT] and PCR was evaluated. Blood samples of sixty ownership dogs [/= 1/320] in DAT while seven of the 60 [11.66%] samples were positive by FML-ELISA. Nine out of 60 [15%] buffy coat samples showed a band about 680 bp indicative of L. infantum in PCR. Three out of 60 dogs had Kala-azar symptoms and were positive by PCR and FML-ELISA, while two of these three dogs had antibody titers >/= 1/320 in their serum samples. The sensitivity and specificity of FML-ELISA for the detection of CVL in both symptomatic and asymptomatic dogs were found to be 77.8% and 100%, respectively. Considering the acceptable sensitivity and high specificity of FML-ELISA, use of this serological method can be recommended for epidemiological surveys of CVL

Zoonotic cutaneous leishmaniasis in Shiraz, Southern Iran: a molecular, isoenzyme and morphologic approach

Cutaneous leishmaniasis [CL] with diverse clinical manifestations is prevalent and remains a major public health problem in Iran and its incidence has been doubled over the last decade. The present study is about the potential role of rodents in the epidemiology of CL in Kharameh district in Shiraz, Southern Iran. From April 2004 to April 2005, a total of sixteen rodents were collected in live traps from the endemic area of CL in Kharameh district in Shiraz. Evans medium was used for culture. Specific polymerase chain reaction and isoenzyme electrophoresis methods were performed to characterize the parasite. The rodent species were Tatera indica. Three samples from Tatera indica were found positive [2 males and 1 female in Kafdehak and Sejel-Abad villages] for L. major. Macrophages in the bone marrow of femoral bone were infected with the amastigote form of the parasite. It seems that T. indica is the reservoir host for CL in Kharameh [a district in Shiraz, Southern Iran]. It was shown that the bone marrow of the rodents is the tissue of choice for light and ultrastructural studies of L. major