Promising antileishmanial effectiveness of doxorubicin and Doxil against Leishmania major: An in vitro assay

OBJECTIVE@#To evaluate the effect of doxorubicin and its pegylated liposomal formulation (Doxil, Caelyx) on in vitro susceptibility of promastigote and amastigote stages of Leishmania major.@*METHODS@#Throughout in vitro assays the IC was calculated in the promastigotes and amastigotes forms in J774 macrophage cell line. Also as cytotoxicity in J774 cell line macrophages.@*RESULTS@#Doxorubicin and Doxil showed the same activity against promastigote form with IC values of 10.49 μg/mL and 9.63 μg/mL, respectively. Similarly, the amastigote stage was susceptible at concentration of at least 1 μg/mL when compared to positive control (P < 0.0001). Also, cytotoxicity assay against macrophage revealed no toxicity on the host cells at IC concentrations.@*CONCLUSIONS@#Our findings demonstrated the efficacy of both doxorubicin and its pegylated liposomal formulation on L. major at low concentrations. Further researches are needed for evaluating the safety of drugs in animal model particularly as topical formulation.

Promising antileishmanial effectiveness of doxorubicin and Doxil against Leishmania major: An in vitro assay

Objective To evaluate the effect of doxorubicin and its pegylated liposomal formulation (Doxil, Caelyx) on in vitro susceptibility of promastigote and amastigote stages of Leishmania major. Methods Throughout in vitro assays the IC

Co-infection with Enterobius vermicularis and Taenia saginata mimicking acute appendicitis

In this report, we describe an unusual case of verminous appendicitis due to Enterobius vermicularis and Taenia saginata in a 29-year-old woman from Iran. The histopathological examinations and parasitological descriptions of both worms found in the appendix lumen are discussed. The removed appendix exhibited the macroscopic and microscopic features of acute appendicitis. Antihelminthic therapy was initiated with single doses of praziquantel for the taeniasis and mebendazole for the enterobiasis, and the patient was discharged

Lupoid leishmaniasis among the known cases of cutaneous leishmaniasis in Herat Province, western Afghanistan

Lupoid cutaneous leishmaniasis [LCL] is an uncommon form of chronic cutaneous leishmaniasis, which is mostly caused by Leishmania tropica in the Old World and has a high incidence throughout early life. Between 2012 and 2013, patients with active lesions suspected to be cutaneous leishmaniasis [CL] were examined. Diagnosis was performed through a combination of methods, i.e., clinical examination, direct smears and kDNA polymerase chain reaction [PCR]. Overall, 162[4.2%] subjects, through clinical examination and PCR confirmation alone, were diagnosed as having LCL, with the duration of the lesions varying from 2 to 5 years. Most [85.8%] of the subjects with LCL were <20 years of age. No amastigote was found in direct smears. Moreover, direct PCR on the negative smears for identifying Leishmania provided a specificity of 100%, and the species was identified as Leishmania tropica using specific kDNA PCR. Performing PCR on skin smears appears to offer a valuable method for the diagnosis of LCL because it is highly specific and sensitive, especially for clinical correlative studies

Molecular identification of ovine Babesia spp. in north of Iran

Babesia is a blood-tissue parasite, which is transmitted by hard ticks from Ixodidae family. The parasite is the cause of babesiosis among small ruminants, cattle, human, dogs and other animals. Babesia is one of the main fatal factors among livestock in endemic regions such as Iran. The aim of this study was to identify Babesia spp infection using microscopic and molecular methods among small ruminants in Mazandaran and Golestan provinces, northern Iran, during 2011-2012. In this study, a total of 220 sheep and goats were selected from 22 flocks in different regions of these provinces and blood samples were taken from their ears. The samples were transferred to the laboratory. Then thick and thin smears were prepared, stained with Geimsa and examined under light microscope. Standard PCR and semi nested- PCR was performed to differentiate genus of Theileria and Babesia, also identify the species of Babesia. From a total of 220 blood samples [160 sheep and 60 goats], 34 cases [15.4%] showed Babesia infection using microscopic examination. Whereas, 11 cases [5%] were found positive for Babesia spp using standard PCR. Also, two positive cases were showed mixed infection with Theileria spp. In addition, two microscopic negative samples were positive by PCR assay. Using semi nested- PCR, Babesia ovis [n=10] and B. motasi [n=1] were detected. Our results shows ovine babesiosis is common in the Northern provinces of Iran. Moreover, Babesia ovis is the main causative agent of ovine babesiosis in northern Iran. The relatively high prevalence of Babesia infection in sheep and goats indicates the epizootic stability status of babesiosis in the northern part of Iran

[Characterization of Leishmania isolated from Kala- azar infected patients in Kermanshah using PCR]

Visceral leishmaniasis [VL] or Kala-azar is an important parasitic disease caused by protozoan parasites of the genus Leishmania including L. donovani and L. infantum. Some evidences show that VL is present in some areas of Kermanshah province and this study aimed to characterize the causative agent of VL in this region. Bone marrow sample was obtained from 9 VL patients from Pediatric Hospital in Kermanshah. DNA was extracted from the microscopic slides and was checked by specific PCR to find out the species of the parasite. To do that, a segment of mini circle kinetoplast DNA was amplified, using LINR4 and LIN17 primers. All samples produced a 720 bp band in PCR reaction. Products of PCR were evaluated by electrophoresis using 1.5% gel agarose. Anti Leishmania antibody was detected in all patients except one case by IFAT. Examination of bone marrow smears demonstrated numerous amastigotes of Leishmania [Leishman bodies] in the samples. The isolates were compared with reference strains and revealed that all isolates were L. infantum. Our findings showed that L. infantum is the most common VL in Kermanshah and one-step PCR reaction may be suitable method to detect species' characterization. Further study is necessary to explore other aspects of VL including seroprevalence and parasite reservoirs in this region

[Application of superior enzymatic systems for characterization of causative agents of visceral leishmaniasis [kala-azar] in Iran using isoenzyme electrophoresis]

Visceral leishmaniasis [Kala-azar] is one of the parasitic zoonotic diseases which is caused by Leishmania donovani complex parasites. Thus, the aim of this study is the application of different enzymatic systems for discrimination species and strains visceral leishmaniasis agent. In this experimental study, reference strains of Leishmania infatum, Leishmania major, Leishmania tropica in addition, the leishmania parasites isolated from bone marrow of subjects and internal organs of dogs infected to visceral leishmaiasis [VL] were inoculated on RPMI + FBS 10% medium for mass cultivation. Then, using electrophoresis on polyacrilamid gel, six enzymatic systems including GPI, PGM, MDH, G6PD, 6PGD and NH2 were examined in order that identification of species and strains of the isolates and thus finding the appropriate enzymatic systems for discrimination of these compared with reference strains. Isoenzymatic profile of six enzymatic systems mentioned above for these isolates were compared with reference strains and also relative migration were calculated. Finally, the results were showed that only five enzymatic systems, except 6PGD, had discriminated ability of different species. In the present study, GPI and G6PD enzymes had the most heterogeneity while NH2 enzyme had the most homogeneity. Moreover, PGM, GPI and MDH were highly active enzymes

Comparison of serological methods [ELISA, DAT and IFA] for diagnosis of visceral leishmaniasis utilizing an endemic strain

The causative agent of visceral leishmaniasis [VL] in Iran is Leishmania infantum [L. infantum] [Mediterranean type] and its major reservoir host is the dog. To compare the serological methods including direct agglutination test [DAT], indirect immunofluorescent-antibody test [IFA] and enzyme-linked immunosorbent assay [ELISA] for serodiagnosis of endemic strain of L. infantum. 61 blood samples from VL patients referred to Shiraz hospitals and 49 blood samples from control group were collected. Native strain of the parasite isolated from a VL patient from the region was cultured and characterized. Antigens from this L. infantum parasite were used in ELISA and IFA system. Anti-Leishmania antibody was detected in 43 [70.5%], 49 [80.3%] and 51 [83.6%] cases using DAT, IFA and ELISA, respectively. Based on these results, sensitivity and specificity of DAT was found to be 70.5% and 100%, respectively. Sensitivities of IFA and ELISA in diagnosis of VL were 80.3% and 83.6% and their specificity was 90.5%. Results of this study showed that DAT and ELISA have the highest specificity and sensitivity in diagnosis of VL. DAT is a simple, cost-effective and field applicable test. Thus, it can be recommended for early and accurate diagnosis of VL, especially in regions where malaria, brucellosis and tuberculosis are prevalent