Comparison of agar dilution, broth dilution, cylinder plate and disk diffusion methods for evaluation of anti-leishmanial drugs on Leishmania promastigotes

Leishmaniases are a group of diseases caused by Leishmania parasites. Growing of drug unresponsiveness in leishmaniasis patients necessitates the development of new drugs and accordingly a suitable assay is needed for evaluation of any modalities. The aim of this study was to compare four drug assays methods, agar dilution, broth dilution, cylinder plate and disk diffusion, for evaluation of anti-leishmanial drugs on Leishmania promastigotes, using glucantime as a currently available drug for treatment of leishmaniasis. For broth dilution method, different concentration of glucantime was added to the parasite culture [promastigotes of Leishmania], while in cylinder plate method wells were punched in agar gel and filled with different concentration of drug and zone of inhibition was measured in each well. In disk diffusion method, the parasites were cultivated on the surface of agar; filter paper disks were enriched with various concentration of glucantime and were placed on the surface of agar. In agar dilution method, various concentrations of drug were incorporated onto blood agar and the parasites were cultivated on the surface of the agar. A direct correlation was found between the drug concentration and size of inhibitory zones in cylinder plate and disk diffusion methods. These two drug assays methods provided much better performance in comparison with broth and agar dilution methods. Cylinder plate and disk diffusion methods seem to be acceptable methods for susceptibility testing of anti-leishmanial compounds on Leishmania promastigotes

Establishment of a modified in vitro cultivation of protoscoleces to adult echinococcus granulosus; an important way for new investigations on hydatidosis

Echinococcus granulosus, a zoonotic cestode parasite, causative agent of hydatid cyst is endemic in many parts of the world including the Middle East. Study on different aspects of this parasite is very important and valuable. However, working with adult worms which their habitat situated in the small intestine of canids, is dangerous and risky. Achieving such risky situation needs a controlled condition which is cultivation of the organisms in the laboratory. In this regard, cultivation of E. granulosus protoscoleces leading to adult worms was established in the laboratory for the first time in Iran. Under aseptic conditions a number of protoscoleces were cultivated in diphasic S.10E.H medium using CO2 incubator to produce adult worms. Different forms of parasites including pre-segmentation stages [PS1 - PS4] and segmentation stages [S5-S8] and developing stages in segmented worms [S10-S11] were observed and evaluated in these medium. Finally adult worms contained four proglottids with a large and distinct genital pore were observed 50-55 days post cultivation. These parasites do not produce fertile eggs and conclusively do not have risk of hydatid disease transmission to the researchers. The mentioned method for producing E. granulosus adult worms can open a new window for researches and facilitate working on different aspects of hydatidosis especially for diagnosis, protection and treatment studies

Helminthic infections of laboratory animals in animal house of Shiraz University of medical sciences and the potential risks of zoonotic infections for researchers

Different parasitic diseases may be transfered from laboratory animals to human [zoonoses]. The current study was designed to determine the helminthic infections in animal house of Shiraz University of Medical Sciences to prevent the possible transmission of zoonotic ones from laboratory animals to the staff and researchers. Sixty laboratory animals including mouse, rats, Guinea pigs and rabbits were randomly selected and examined for any helminthic infections. 83.33% rats were infected with Syphacia muris and Aspiculuris tetraptera, 100% of Guinea pigs were infected with Paraspidodera uncinata, rabbits were infected with Passalurus ambiguous [40%], inbred BALB/C mice were infected with Hymenolepis nana [50%], Aspiculuris tetraptera [90%] and Syphacia obvelata [90%], outbred BALB/C mice were infected with Hymenolepis nana [50%], Aspiculuris tetraptera [90%], Syphacia obvelata [90%] and C57BL/6 mice were infected with Hymenolepis nana [66%], Aspiculuris tetraptera [100%] and Syphacia obvelata [100%]. Our study was revealed minimum and maximum infection frequency in rabbits and guinea pigs respectively. It seems that low and unsuitable space of infected animals in mentioned research center was the main cause of distribution of infection among rats and mice in Animal House of Shiraz University of Medical Sciences

case of gingival myiasis caused by Wohlfahrtia magnified

A gingival myiasis in a four years old mental retarded boy with anorexia and weight loss is presented from southern part of Iran. Entomological studies on larvae showed the larvae as Wohlfartia magnifica which is a rare causative agent of gingival myiasis

Study on the genomic diversity of hymenolepis nana between rat and mouse isolates by RAPD-PCR

Hymenolepis nana is a common parasite of rodents as well as human intestine. This parasite has been reported from all over the world, including Iran. The infection rate has been reported up to 40% in some areas. The infection has various clinical manifestations. The parasite could establish severe hyperinfection in patients with immune deficiency. Regarding the rodents as hosts of the parasite, the infection may disseminate through these hosts to the nature. As H. nana is a zoonoses, phylogenic study of this parasite is of particular importance. Considering these criteria, the genomic diversity of 16 H. nana with the origin of Shiraz and Tehran were studied among the worms of mice and rats by RAPD-PCR. Genomic DNA extracted from individual worms by proteinase K method and three oligonucleotides primer [ABl-17, UBC-358, UBC-387] were used for RAPD-PCR. Similarity index were calculated by Nei and Li method. Data were analysed using UPGMA analysis and dendrograms were obtained by group average method with 100 bootstrapping analysis. The range of genomic similarity determined among specimens by ABl-17 primer was 48.3-90%, by UBC-358 primer 55-87% and by UBC-387 primer 53-97%. Regarding our data and genomic similarity indexes, various isolates were found in both specimens of rats and mice. However no differences were obtained between H. nana from rat or mouse isolates by these primers. The results showed that it is not possible to divide the isolates into two distinct groups based on their origin as Tehran and Shiraz