فهرست مطالب farnoosh jameie

Iran is one of the endemic areas of Mediterranean Visceral Leishmaniasis, a disease caused by Leishmania infantum. In this work, we examined whether Proteína quimérica 10 (PQ10) recombinant protein is suitable for immunological diagnosis of human visceral leishmaniasis.

The study was carried out in Tarbiat Modares University during 2016- 2018. The coding sequence of PQ10 recombinant protein was sub-cloned in pET28 expression vector and was commercially synthesized by GENERAY Biotechnology, China. Sequencing with proper primers was done, the expression, optimization of expression and protein purification were performed, and the purified recombinant protein was confirmed by western blot. The efficacy of PQ10 for serodiagnosis was evaluated with 50 positive and 50 negative serum samples, which confirmed by the direct agglutination test and collected from individuals living in the visceral leishmaniasis endemic areas of Iran. ELISA was performed with the PQ10 recombinant protein.

The 95% CI sensitivity of ELISA that was evaluated with sera from naturally infected individuals was 84%. The 95% CI specificity value of the ELISA determined with sera from healthy individuals (50 serum samples) and from individuals with other infectious diseases was 82%. The 95% CI positive predictive value (PPV) and negative predictive value (NPV) were exterminated 82.35% and 83.67%, respectively.

We have used a recombinant synthetic protein to improve serodiagnosis of human visceral leishmaniasis. PQ10 could be useful for diagnosis of asymptomatic cases, as well as in the early phase of infections.

Diagnosis of Cryptosporidium species based on morphological characteristics is very limited and has no taxonomic value alone, and therefore the use of molecular methods removes these limitations to some extent.

The aim of this study was to determine the predominant Cryptosporidium genotypes in calves with diarrhea.

Study were conducted in calves aged less than 3 months for a period of 2 years. During the study period, 160 dung samples were collected from neonatal calves and examined first microscopically and then by molecular techniques. Stools were analyzed for the presence of Cryptosporidium oocysts by Sheather's Sugar Flotation Solution followed by Ziel-Neelsen staining method. DNA of parasite was extracted and multiplex nested-PCR protocol basis on the 18srRNA were done to identify three cattle-adapted species (C. andersoni, C. bovis and C. ryanae) plus the zoonotic species C. parvum.

110 fecal samples were collected from livestock in Alborz province and 50 fecal samples were collected from livestock in Shahroud city. Of the 160 animals examined, 90 were female and 70 were male. In total, out of 160 animals examined, 85 cases (53.12%) had diarrhea, of which 55 cases (34.37%) were positive using Ziel-Neelsen staining. Since all positive cases were related to diarrhea samples and related to calves under one month old, a significant relationship was observed between diarrhea status and the presence of this parasite (p < /em><0.05). In terms of seasonal distribution, no difference was observed in the rate of diarrhea and positive parasitic cases. The presence of 305 bp band in all Ziel-Neelsen positive samples confirmed the presence of C. parvum in all samples.

Neonatal calves are more likely to be infected with Cryptosporidium parvum, as confirmed by the present study.

Canine Leishmaniosis due to Leishmania infantum is a major global zoonosis, potentially fatal to humans and dogs.Blood samples were taken and collected from cephalic or saphenous vein from 85 stray dogs of different ages, breeds, and both genders that were registered in the Karaj municipality's dog camping and 60 stray dogs that were registered in the Kohsar district. After separation of blood sera, anti-Leishmanial antibodies were detected by direct agglutination test (DAT). Out of 85 stray dogs samples from the Karaj municipality's dog camping, 38(44.7%) were males and 47(55.3%) were females and out of these, 4(4.7%) of them were positive for visceral Leishmaniasis that three of them were male and one was female. All of the seropositive dogs were symptomatic. Out of 60 stray dogs samples from the kohsar district, 14(23.3%) of them were positive for visceral Leishmaniasis that 6(10%) of positine cases were males and 8(13.3%) were females .Among the seropositive dogs in this regions, 4(6.6%) cases were asymptomatic and 10(16.66%) cases were symptomatic. there was not any significant difference between season of sample collection .Visceral Leishmaniasis is a zoonotic disease; therefore, the need for continuous surveillances on the prevalence of it in our communities is an essential work as a control strategy. There is a need for more additionalserological and molecular studies for exact determination of the presence of these parasites.

Acanthamoeba is a ubiquitous amphizoic organism which can cause lethal diseases such as granulomatous amoebic encephalitis and unfortunately, the infection has now increased in the world. The aim here was to evaluate in vitro anti-Acanthamoeba properties of crude aqueous and ethanolic extracts of Myrtus communis. In this experimental research, a clinical isolate of Acanthamoeba was cultured and genotyped. The aqueous and ethanolic extracts of Myrtus communis were prepared. Then, various concentrations of Myrtus communis extracts (1.25, 2.5, 5, and 10 mg/ml) were tested at three different times (24, 48 and 72 hr) on trophozoites and cysts of Acanthamoeba in vitro. The viability of trophozoites or cysts was tested by trypan blue method. Unstained (viable) and stained (nonviable) parasites were evaluated by counting with a neobar lam. The percentage of viablity of trophozoites and cysts after adding ethanolic extract of Myrtus communis was 0% and 8.62%, respectively. Moreover, at 10 mg/ml concentration of aqueous extract of Myrtus communis, 0% trophozoites and 31.10% cysts lived after 72 h. This extract can be used as a safe anti-Acanthamoeba agent against trophozoites and cysts of Acanthamoeba and further investigations are recommended to show the effects of this plant as an antiparasitic drug in animal models and volunteer infected people.

The protozoan parasites of the genus Leishmania are the causative agents of various clinical diseases. Different methods of cultivation of Leishmania parasites are available. In the present study, the efficacy of the LB broth with rabbit lyophilized anti-sheep red blood cell haemolysin was evaluated in the cultivation of promastigotes of Leishmania major. Conventional LB broth medium was prepared and autoclaved for 15 min at 121°C and then lyophilized rabbit anti-sheep cell haemolysin was added at 1-10% final concentrations. The efficacy of the medium was evaluated by assessing the growth ability and replication patterns of the promastigotes of Leishmania major. Medium with 1-10% lyophilized rabbit haemolysin supported the growth of the parasites and can be used for cultivation of Leishmania parasites with acceptable In vivo infectivity for research purpose. The ability of the parasites to survive and proliferate in the presence of lyophilized rabbit haemolysin indicates that this material is a good nutritional source. This study opens a new way to make low-cost medium that can be used in cultivation of Leishmania parasites.

Cystic echinococcosis (CE), as a zoonotic disease cause to health threat and economic losses. Despite implemented control programs, few countries have been able to decrease or eliminate this infection. Vaccination of the intermediate host offers an additional strategy to control the parasite transmission and EG95 antigen is considered more than the others in the vaccine issue. According to the high protection induced by the EG95 recombinant vaccine, this study was designed to construct recombinant plasmid formulation of EG95 antigen.
In 2015, the Echinococcus granulosus eggs were recovered from an infected dog in Parasitological laboratory of Tarbiat Modares University in Tehran, Iran. Following hatching, the oncospheres of E. granulosus were activated to increase the presence of the desired mRNA. The extracted mRNA was transcribed to the cDNA which used as template in RT-PCR. Then the EG95 gene cloned into pET28a vector and the recombinant plasmids expression was investigated in prokaryotic and eukaryotic cells.
The recombinant plasmid encoding EG95 antigen was successfully constructed and identified by PCR, restriction enzyme digestion and sequencing. In vitro expression of the EG95 antigen was confirmed in prokary­otic and eukaryotic systems by SDS-PAGE and western blotting analysis.
Because of potential advantages of DNA vaccines, including ability to induce long-term immune responses, low production cost and stability in different temperatures, this study carried out to construct the EG95 gene into a vector. This recombinant vector can be evaluated in further studies as a DNA vaccine may provide new prospects for the development of a vaccine against cystic hydatid disease.

Les espèces d’hépatozoonose sont des protozoaires infectant les animaux tels que les oiseaux, les reptiles, les amphibiens et les carnivores. Bien que l’hépatozoonose canine ait été rapportée en Iran, elle na pas encore été détectée par des méthodes moléculaires. Lobjectif principal de cette étude était le diagnostic moléculaire de l’hépatozoonose canine par la méthode PCR et la détermination de sa séquence nucléotidique. A cet effet, 104 échantillons de sang provenant des chiens de la ville de Meshkinshahr (Province Ardabil) ont été recueillies et les ADNs ont été extraites à l’aide du kit DNG-PLUS. Ensuite le gène SrRNA 18 a été amplifié par PCR conventionnelle afin de déterminer sa séquence. Une bande denviron 897 pb a été identifiée dans les échantillons positifs et 24 des 104 échantillons analysés (23.7%) étaient infectés par une hépatozoonose canine. Ce pourcentage dinfestation chez les chiens de la province Ardabil est relativement élevé. La séquence moléculaire du fragment étudié était 99% similaire à celles enregistrées dans la banque de gènes. Cette étude représente le premier rapport sur la détection de l’hépatozoonose canine en Iran.

Various Leishmania species can cause human infection with a spectrum of clinical manifestations. The current treatments are unsatisfactory; and in absence of a vaccine there is an urgent need for effective drugs to replace those currently in use. In the present study, the effect of direct current electricity in combination with selenium and silver nanoparticles on the healing of leishmanial lesions has been investigated. In this experimental study, 24 Balb/c mice were divided into four groups and infected experimentally with L.major. Then silver nanoparticles with a concentration of 250 mg/kg and selenium nanoparticle with a concentration of 125 mg/kg were injected inter-lesion, and simultaneously 0.5 mA DC induction was applied directly into the wound. Finally, the lesion size and mice body weight changes were measured during five weeks. The lesions were found to be smaller in the group treated with DC and silver nanoparticles than the group treated with DC and selenium nanoparticles (P < 0.05). Two treated groups showed significant differences with both negative control group and the group treated with Glucantime (P < 0.05). The results showed that selenium and silver nanoparticles with direct current electricity promotes wound healing in mice.