Molecular Identification and Genotyping of Theileria Orientalis Type 3 (Buffeli) Isolated from Cattle Using Nested-PCR Assay in Guilan Province, Iran.

Protozoan parasites of the genus Theileria are tick-borne parasites that have been found in many species of mammals. More than a dozen species of Theileria have been found in cattle, water buffalo, sheep, and goats. Theileria orientalis is a non-pathogenic blood protozoan parasite that was detected and identified during a regular investigation of piroplasmida infection in indigenous cattle in the spring of 2019 in Northern Provinces of Iran. In total, 92 blood samples were collected from different areas of Guilan and Mazandaran Provinces, Iran during the spring. The Giemsa stained blood smears did not show any parasitic infection; however, T. orientalis was identified by 18S rRNA gene polymerase chain reaction (PCR) and DNA sequencing. The specific sequenced DNA for T. orientalis was registered in GenBank under the accession number MN453385. The partial 18S rRNA gene sequence of the obtained DNA showed 100% nucleotide identity with reference sequences for the T. orientalis that have been registered from Europe, Africa, and Asia. Additionally, molecular phylogenetic studies have shown that T. orientalis Iran GC98-01 isolate belongs to nonpathogenic T. orientalis type 3 (buffeli). In this study, the indigenous Bos indicus cattle were detected as asymptomatic carriers of Theileria spp. infection. Here, we identified and genotyped T. orientalis for the first time as T. orientalis type 3 (buffeli) in Iran using molecular phylogenetic analysis and registered the 18S rRNA gene sequence of the T. orientalis GC98-01 isolate in GenBank. Moreover, rare T. annulata infection was detected in cattle using semi-nested PCR in Mazandaran (Miankaleh peninsula). The T. orientalis can be differentiated from other Theileria and Babesia haemoprotozoan parasites by specific molecular assays.

Molecular detection of Theileria spp. and Babesia ovis Infection in Sheep in Baneh, Iran.

The purpose of the present study was to investigate the Theileria and Babesia infection in sheep using polymerase chain reaction (PCR) assay in Baneh, Iran. Theileria and Babesia are apicomplexan parasites that have both vertebral and invertebrate hosts. These protozoa, which are transmitted by tick vectors, are considered to be the most important causes of parasitic diseases in Iran.The detection methods of Babesia and Theileria spp. are morphological examination, serology tests, and more recently, molecular assays, such as PCR. In this study, a total of 66 blood samples were collected and analyzed using specific primers for Theileria annulata, T. ovis, T. lestoquardi, and Babesia ovis. Two PCR methods were used, namely semi-nested PCR and competitive PCR. Based on the results of the PCR assay of 66 sheep blood samples, B. ovis, T. ovis, T. lestoquardi, and T. annulata were detected in 57 (86.4%), 28 (42.4%), 0, and 16 (24%) cases, respectively. Detection of low levels of protozoan infection with high morbidity in the tested animals shows their status as a carrier that keeps the infection in the region and extends the protozoan life cycle. Another important factor is the geographical situation of Baneh as a border city since the hemoprotozoan infection is present in this region. Moreover, piroplasmida infection was found in Iraq and other neighboring provinces. Therefore, animal husbandry in Baneh is at the risk of infection with Babesia and Theileria. The collected data in this study are useful for reaching a better understanding of the epizootiology of theileriosis and babesiosis, in order to control and prevent the diseases in this region.

Rapamycin delays salivary gland atrophy following ductal ligation.

Salivary gland atrophy is a frequent consequence of head and neck cancer irradiation therapy but can potentially be regulated through the mammalian target of rapamycin (mTOR). Excretory duct ligation of the mouse submandibular gland provokes severe glandular atrophy causing activation of mTOR. This study aims to discover the effects of blocking mTOR signaling in ligation-induced atrophic salivary glands. Following 1 week of unilateral submandibular excretory duct ligation: gland weights were significantly reduced, 4E-BP1 and S6rp were activated, and tissue morphology revealed typical signs of atrophy. However, 3 days following ligation with rapamycin treatment, a selective mTOR inhibitor, gland weights were maintained, 4E-BP1 and S6rp phosphorylation was inhibited, and there were morphological signs of recovery from atrophy. However, following 5 and 7 days of ligation and rapamycin treatment, glands expressed active mTOR and showed signs of considerable atrophy. This evidence suggests that inhibition of mTOR by rapamycin delays ligation-induced atrophy of salivary glands.

Detection and Identification of Toxoplasma gondii Type One Infection in Sheep Aborted Fetuses in Qazvin Province of Iran.

BACKGROUND: The aim of this study was to apply the nested-PCR and bioassay methods in detection and genotyping of Toxoplasma gondii infection in provided sheep aborted fetus samples from Qazvin Province of Iran. METHODS: Eighteen sheep aborted fetal samples were studied by nested-PCR-RFLP, histopathological observation and microbiological assay. Bioassay in mice was carried out by inoculating the brain samples intraperitoneally. RESULTS: The results demonstrated the frequency of 66% infected sheep aborted fetal samples with T. gondii type one. Although we could not isolate any parasite from inoculated mice even after three passages, but it was confirmed histopathologically formation of cyst like bodies in prepared mice brain sections. CONCLUSION: The results of the performed nested-PCR and formation of brain cyst in inoculated mice exhibited that T. gondii type one infection might be considered as one of the major causative agents for abortion in ewes.

Synthesis, characterization and evaluation of computationally designed nanoparticles of molecular imprinted polymers as drug delivery systems.

The aim of the present study was to prepare nanoparticles of molecular imprinted polymers (MIPs) with high loading capacity for naltrexone as template drug. To achieve this goal, a computational protocol was employed to select the most appropriate monomer for MIP preparation. Density functional theory (DFT) method at the B3LYP level of theory in conjugate with the 6-31+G(d) basis set was used to evaluate the extent of interaction between naltrexone and a small library of frequently used vinylic monomers. The results revealed that acrylic acid (AA) and methacrylic acid (MAA) can be considered as suitable monomers. To select the best monomer, two MIPs with AA and MAA monomer were synthesized and their loading capacity, selectivity and release profile were evaluated. The experimental results showed that the MIPs synthesized using AA (MIP-AA) exhibited a surprisingly high loading capacity to naltrexone (75mg of drug/g of MIP) compared to MIP-MAA (34mg of drug/g of MIP). In vitro release dynamics of the drug from MIPs was also investigated and modeled. It was found that non-Fickian-type diffusion mechanism was responsible for drug release. The results can lead to the conclusion that MIPs designed by computational approach can be considered as promising candidates for drug delivery systems.

Prevalence and risk factors of hepatitis A among blood donors in Qazvin, central Iran.

INTRODUCTION: The hepatitis A virus (HAV) is a major global health problem, especially in developing countries. Although children aged 5-14 years are the most infected age group, all age groups are equally affected in an HAV epidemic. The aim of this study was to determine the prevalence and related risk factors of HAV among blood donors in Qazvin, central Iran. METHODS: A cross-sectional study was performed in April 2008 on a total of 351 blood donors aged 17 to 60 years. Information on demographic variables and probable risk factors was recorded. The blood samples were investigated for antibodies to HAV (HAV Ab). Data was analysed using the t-test, chi-square test and Fisher's exact test to examine the factors related to HAV Ab positivity. RESULTS: Out of the 351 blood donors, 49.0 percent (n is 172) were regular blood donors, 94.0 percent (n is 330) were male and 71.0 percent (n is 248) were married. HAV Ab was found in 94.9 percent (n is 333) of the total sample. The prevalence of HAV Ab was higher among married and older participants. It was lower among those with a high school or vocational level of education (p is less than 0.05) than among those with a lower level of education. CONCLUSION: This study reveals a high prevalence of HAV Ab among adult blood donors in Qazvin, which indicates a high level of childhood contact with HAV in the area.