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Aim: Cattle is one of the main sources of food supply chain for humans in most countries. The present study aimed to identify the infection rates for Toxoplasma gondii, Neospora caninum, and Sarcocystis spp. by polymerase chain reaction (PCR) method in native slaughtered cattle in Kashan, central Iran. Materials and Methods: Totally, 159 diaphragm, esophagus, and muscle samples (53 samples of each) were collected from native beef cattle from Kashan slaughterhouse, central Iran. The genomic DNA was extracted, and PCR method was used separately for detection of N. caninum, Sarcocystis, and T. gondii species using specific primers. Finding: Sarcocystis was found in 84.9% of muscles, 83% of esophagus, and 84.9% of diaphragm samples. Mixed infection (Sarcocystis cruzi–Sarcocystis hominis) was the most common infection, followed by S. cruzi and S. hominis. Sarcocystis hirsuta was not detected in any samples. T. gondii was detected only in three (5.7%) out of the 53 muscle tissues samples of cattle. N. caninum was found in 18.9% of muscles, 24.5% of esophagus, and 28.3% of diaphragm samples. One of the cattle had coinfection to Neospora, Toxoplasma, and Sarcocystis in muscles simultaneously. There was no statistically significant difference between infection rates and age as well as sex in each organ. Conclusion: This study revealed a low prevalence rate of T. gondii, but a high prevalence of infection to N. caninum and S. cruzi or mixed infection of S. cruzi with S. hominis among slaughtered cattle. Prevention measures such as keeping away dogs from cattle grazing are recommended.

Aims: Hydatidosis is an important zoonotic disease that is caused by a tiny tapeworm, namely Echinococcus granulosus. In this study, three polymerase chain reaction (PCR)-based methods, including, high resolution melting (HRM) analysis, DNA sequencing, and PCR-restriction fragment length polymorphism (RFLP) have been used for genotype the identification of E. granulosus isolates from dogs and camels in Zarinshahr and Najafabad, Isfahan province, Iran. Materials and Methods: A total of 200 adult worms of 40 dogs and 51 samples of camel hydatid cysts were examined. Molecular characterization of isolates was performed using HRM assay, sequencing of DNA, and digestion Rsa1 pattern coding for the mitochondrial cox1 gene. For analysis of the HRM melting curve, we used the Tm within the range of 77.50°C–79.23°C. Results: HRM analysis revealed that 72.5%, 15%, and 12.5% dog's genotypes and 41.17%, 21.56%, and 35.29% camel genotypes were G1, G3, and G6, respectively. PCR-RFLP analysis, spare parts 310 bp and 138 bp of cox1 that shows the G1 genotype in all of the isolates. Sequence analysis as well as HRM assay was confirmed genotypes of G1, G3, and G6 in camels and dogs. Based on three methods of the cox1 gene the dominant genotype was G1. Conclusion: The PCR-RFLP only identified the G1 genotype, whereas the HRM analysis, as well as DNA sequencing, were detected three genotypes G1, G3, G6, therefore, these two methods have enough accuracy for the determination of genotypes of E. granulosus. This information leads to a better understanding of the biological characteristics of E. granulosus genotypes in Iran and shows the camel as a source of human hydatidosis.

Aims: The control of fascioliasis has depended on the utilization of a predetermined number of anthelmintic drugs. Nonetheless, the resistance of antifasciolid is presently boundless, and there is a need for potential drug properties of medicinal plants as new medications against Fasciola spp. to interrupt the parasite transmission. Materials and Methods: This investigation is meant to assess the potential anti-fasciolicide impacts of Zingiber officinale roscoe hydroalcoholic extract against the Fasciola miracidia. The eggs of Fasciola hepatica were gathered from the livers contaminated goats and sheep and washed 20 times with chlorine-free water, and afterward incubated at different pH, temperature, and light density for embryonic development. Concentrated miracidia of Fasciola spp. were incubated at different concentrations (2, 5, and 10 μg/mL) of hydroalcoholic extract of Z. officinale at different time intervals. The data were analyzed using the ANOVA statistical test. Results: The extract at different concentrations showed antifasciolid effects in comparison to triclabendazole. This braking action was dose-proportional and further related firmly to the disclosure time ( P < 0.001). In concentration of 10ug/ml extract killed the parasites at 105 ± 3 s ( P < 0.001). Conclusion: In vitro antimiracidia activities of the hydro alcoholic extract of Z. officinale was satisfactory and potent fasciolicide effective, however, in vivo impact of this extract, remains for extra assessment. In this manner, these therapeutic plant extracts might be seen as confident origins of bioactive composites that could be matured against miracidia. This is the main work to evaluate the potential enemy of fasciolicide impact of certain plants against Fasciola miracidia.

Introduction: Free –living Amoeba (FLA) are microorganisms that are abundant in the nature. Some genus of these protozoa are opportunistic and cause granulomatous encephalitis and lung infections in immunocompromised people such as cancer patients. Due to the little information about the status of infection of these parasites in the above patients, the present study was performed to determine the prevalence of FLA in cancer patients in Kashan hospitals. Materials and Methods: This cross-sectional study was performed on hundred oral cavity mucosal samples of cancer patients referred to Shahid Beheshti and Yathribi hospitals in Kashan during 2019 to 2020. The samples were cultured onto 1.5% Non-nutrient agar enriched with killed Escherichia coli and examined for the presence of FLA. The rate of contamination with free living amoebae and its relationship with demographic information of patients were analyzed by SPss. 16 Results: Overall, the prevalence of free-living amoeba in oral cavity of cancer patients was 89%. Also, 92.6% of breast cancer and 90.9% of bowel cancer patients were positive for FLA, respectively. The highest rate of FLA contamination was observed in women over the age of 66 years. In addition, the highest rate was observed in 90.9% of illiterate people and the lowest in 76.9% of academics, but the difference was not significant. Conclusion: The rate of free-living amoeba in oral cavity of cancer patients in Kashan were more than the study results of Iran and world. Therefore, health education about transmission and sanitation is recommended for prevention of infection.

The article's abstract is no available.

Recurrence of hydatidosis may results from cyst fluid spillage during surgery. To inactivate the cyst content, different scolicidal agents have been introduced. Nevertheless, novel and more effective treatments are needed due to the associated complications. In the current study, we examined the scolicidal effects of gold nanoparticles on Echinococcus granulosus protoscoleces. Hydatid cysts of sheep liver were collected in this study. The cyst fluid containing protoscoleces was aspirated aseptically. The protoscoleces were exposed to gold nanoparticles at different concentrations (250, 500, 1000, 2000, and 4000 μg/mL). Eosin staining method was applied to determine the viability of protoscoleces at 5, 10, 20, 30, and 60 min. Moreover, cytotoxicity analysis, scanning electron microscopy (SEM) of ultrastructural changes, DNA fragmentation, were performed, and protoscoleces were studied following treatment. The significant scolicidal effects of gold nanoparticles were observed at all concentrations compared to the control group. A total of 4000 μg/mL of gold nanoparticles could destroy 76% of protoscoleces in 60 min. Cytotoxic effects of gold nanoparticle on J774 macrophage cell line, in minimum and maximum concentration (500 and 4000 μg/mL), were 1 and 11%, respectively. After treatment of protoscoleces with gold nanoparticles in different times and concentrations, considerable alteration in size, ultrastructure changes in tegument and shape of sucker, as well as DNA fragmentation of parasite were seen. Based on the results, gold nanoparticles could exert scolicidal effects on hydatid cyst protoscoleces; therefore, they can be applied in hydatid cyst treatment.

Background: Neosuporosis is caused by a parasitic protozoan called Neospora caninum. This parasite can infect different species of domestic and wild animals. Some studies have reported the possibility of human infection with this parasite. The aim of this study was to evaluate the anti-Neospora caninum antibody in raw milk of cattle in Kashan city. Materials and Methods: In this study, 187 milk samples were collected from two industrial farms in Kashan. The breed of all cows was Holstein. Before sampling, the livestock information such as age, race, history of abortion and stillbirth, the daily milk yield was recorded in the prepared forms. Fat of the samples was separated by centrifuge and lactoserums were collected. To determine anti-Neospora caninum antibody in milk samples, an indirect ELISA kit was used. Results: Results showed that 18.8% (CI=18.18±5.53) of the samples were infected with Neospora caninum. No significant relationship was found between the infection with this protozoa and milk production and age of the cows (P=0.84). Also, the results showed that there was no significant relationship between abortion and infection with Neospora. Conclusion: Considering the contamination rate of cow's milk in Kashan, the implementation of control programs in livestock has great importance. Also, considering the possibility of human infection with Neospora, the need for attention to these control programs is more and more evident.

Background: Hydatidosis is one of the most important zoonotic parasitic disease worldwide caused by Echinococcus granulosus. Given the importance of parasite genotyping in disease control, this study was aimed to evaluate the genetic variation of the camel and dog isolates in Isfahan. Material and Methods: This cross-sectional study was done on infected livers and lungs of camels with hydatid cyst (n=51) during 2015-2016, collected from Zarinshahr and Najafabad industrial abattoir and 200 adult E. granulosus worms collected from stray dogs in different geographical regions of Isfahan for the molecular identification of mitochondrial genes (CO1, ND1). The DNA of protoscolices and adult worms were extracted, the PCR and sequencing were done using specific primers. Pairwise sequence alignment was used to identify the genetic similarity between the sequences and the registered ones in Gen bank using Clustalw2 software. Results: The PCR and sequencing of CO1 and ND1 genes on dog and camel samples showed different E. granulosus genotypes, including the common sheep strain (G1), bovid strain(G3) and camel strain (G6) in Isfahan region. The most frequent strains were the common sheep strain (G1). The genetic similarity between both ND1 and CO1 isolates taken from camel, dog and the isolates registered in Gene bank were 30.25-100% and 27.47-100% for camel and dog, respectively. Conclusion: Mitochondrial ND1 and CO1 genes are proper molecular markers for identifying the genetic variation of the camel and dog E. granulosus. The dominant strain of the worm in Isfahan, is the same as the other parts of the country, i.e., the G1 genotype (the most common sheep strain) belonged to the G1- G1 E. granulosus complex.

Background and Aims: The aim of this study is to clarify nitric oxide (NO)-production by spleen and the importance of spleen in malaria infection in murine model. Materials and Methods: Thirty outbred NMRI female mice were divided into four groups, Group I: No intervention (Healthy control), Group II: With splenectomy (Healthy test), Group III: No intervention, Inoculation of contaminated blood (Infected control), Group IV: With splenectomy, inoculation of contaminated blood (Infected test). The Parasitemia was counted every other day through Giemsa stain examination of animal blood. The parasitemia and survival rates, hepatosplenomegaly and body weight were recorded. After terminal anesthesia, plasma and liver/spleen suspensions were assessed by the Griess micro assay for measurement of NO-levels. Results: At the end of the experiment (on day 16), the parasitemia was 26.99±0.46 % among the group of non-splenectomized animals (Group III) compared with 31.25±0.72% among the group of splenectomized animals (Group IV). The average parasitemia among the groups at the end of the experiment was statistically significant (Group III, Group IV: p= 0.0002). Survival rate was statistically significant (p

Background: In Iran, both forms of cutaneous (CL) and visceral leishmaniasis (VL) have been reported; so the accurate species identification of the parasite(s) and the analysis of genetic diversity are necessary. Methods: The smears were collected from lesions samples of 654 patients with CL, who attended local health centers in 12 provinces of Iran during 2013-2015. The smears were checked for the presence of amastigotes by light microscopy. DNA of 648 Leishmania isolates, amplified by targeting a partial sequence of ITS (18S rRNA–ITS1–5.8S rRNA–ITS2) gene. Twenty-five of all the amplicons were sequenced and analyzed with restriction fragment length polymorphism (RFLP) using the Taq1 enzyme. Results: All the smears were positive microscopically. The PCR-RFLP analysis revealed that 176 (27%) CL patients were infected with L. tropica and, 478 (73%) with L. major. The dominant species in all over Iran is L. major. The sequencing results of all CL patients and RFLP analysis confirmed each other. Based on our phylogenetic tree, 25 ITS DNA sequences were grouped into two clusters representing L. major and L. tropica species. Phylogenetic tree derived from the ITS sequences supports a clear divergence between L. major from the other species. Conclusion: Discrimination of Iranian Leishmania isolates using ITS gene gives us this opportunity to detect, identify, and construct the phylogenetic relationship of Iranian isolates.

The most important transmission path of intestinal parasites is the consumption of contaminated water and food. This survey was conducted to determine the prevalence of intestinal parasites among food handlers in Kashan, central Iran. This cross-sectional study was carried on 1018 food handlers who referred to the health centers in order to receive health certificate in 2017–2018. Stool samples of food handlers were collected and examined using direct and formalin-ethyl acetate methods. Results were recorded in information form along with some demographic data such as sex, age. The data were analyzed based on the tests of descriptive statistics by SPSS 16. Of the 1018 individuals examined 851 (83.6%) were men and 167 (16.4%) were female. Infection rate was 10.2% among the population and 9 types of parasites were diagnosed totally. Rate of infection to protozoan intestinal parasites were: Blastocystis sp. 7.17%, Giardia duodenalis 1.2%, Entamoeba coli 1.27%, Endolimax nana 1.08%, Dientamoeba fragilis 0.3%, Iodamoeba butschlii 0.3%, Entamoeba histolytica/E. dispar 0.2%, Chilomastix mesnili 0.1%. Hymenolepis nana (0.1%) was the only intestinal worm that observed. In this investigation, 8.9% of the individual were infected by one parasite, 1.08% by two and 0.2% by 3 or more parasites. This study showed that infection to intestinal helminthic is rare among food handlers in Kashan but the prevalence of protozoan intestinal parasites is high that similar to other regions of Iran. To reduce the rate of parasitic infections and inhibition of their transmission, continuing the process of promoting the health level is recommended.

Neospora caninum and Toxoplasma gondii are two closely related protozoan parasites that have been detected from various species of bird hosts. However, little is known about the prevalence of N. caninum and T. gondii in crows. Hence, we examined the molecular frequency of N. caninum and T. gondii in the brain samples of hooded crows (Corvus cornix) that collected from different public parks of Tehran, Iran by nested-PCR method. We used the primers targeting the Nc5 and GRA6 genes for detection of N. caninum and T. gondii, respectively. From a total of 55 brain samples, 5 (9.9%) and 9 (16.36%) samples were positive for N. caninum and T. gondii, respectively. Sequencing of a N. caninum isolate revealed 95%-100% identity with the deposited N. caninum in GenBank. Genotyping of T. gondii isolates by PCR-RFLP analysis of the GRA6 gene revealed type III genotype in 8 isolates. The results of this study indicate that hooded crows may have a putative role in transmission of N. caninum and T. gondii to canines and felines definitive hosts, respectively.

Background: One of the most important zoonotic helminths in the world is known as Echinococcus granulosus. Different strains of the E. granulosus have been described based on morphological and molecular characterizations, however, there is limited information regarding the characteristics of the phenotypes and genotypes of E. granulosus in Iran. Methods: The present study was prepared to evaluate the phenotypic and genotypic diversity of E. granulosus isolates collected from human, goat, sheep, and cattle based on 19 standard morphometric parameters and mitochondrial and nuclear genes (CO1, ND1, and ITS1) in Kashan, Markazi Province, Iran during 2013–2014. Results: The biometric analysis for the 19 characters revealed that the 19 morphometric values of cattle isolates were exceptionally higher than human, goat, and sheep isolates (P

Background: Based on genotyping study of human isolates of Giardia lamblia; humans are mainly infected by two assemblages A and B. The present study was carried out to determine the sub-assemblages of G. lamblia isolated from food handlers referred to Kashan health centers, central Iran, 2015. Methods: In this cross-sectional study, 3653 stool samples collected from food-handlers that annually refer to health center for getting a health certification and examined microscopically for G. lamblia cyst. Totally, 44 isolates were selected from 47 Giardia positive samples. Cysts were partially purified by the sucrose density gradient method. After freeze-thaw cycles, genomic DNA was extracted using QIAamp Stool Mini kit. A single step PCR-RFLP method was used to amplify a 458bp fragment at the glutamate dehydrogenase (gdh) locus, restriction enzymes BspLI and RsaI were used for distinguish between genotypes A and B and their subgroups. Results: Of 44 isolates, 24(54.5%) were sub-assemblage AII, 9(20.5%) group B including 7(15.9%) BIII and 2(4.6%) BIV sub-assemblage and 11(25%) isolates showed a mixed pattern of AII and B. Sub-assemblage AI was not detected in this study. Conclusion: The higher rate of sub-assemblage AII demonstrated an anthroponotic origin of the infection so infected food-handlers could directly transmit this protozoan to consumers via contaminated food and water. For finding of pattern of transmission and distribution of Giardia assemblages and sub-assemblage, more studies in human and animal population in different regions are necessary.

Little is known about latent infection and molecular characterisation of Neospora caninum in sheep (Ovis aries). In this study, 330 sheep samples (180 hearts and 150 brains) were analysed for N. caninum DNA by nested polymerase chain reaction (PCR) targeting the Nc-5 gene. Neospora caninum DNA was detected in 3.9% (13/330) of sheep samples. The parasite’s DNA was detected in 6.7% of heart samples (12/180) and 0.7% (1/150) of brain samples. No clinical signs were recorded from infected or uninfected animals. Sequencing of the genomic DNA revealed 96% – 99% similarity with each other and 95.15% – 100% similarity with N. caninum sequences deposited in GenBank. To our knowledge, this is the first report on the use of PCR to identify latent neosporosis in sheep in Iran. The results of this study have the potential to contribute to our understanding of the role of N. caninum-infected sheep in the epidemiology of neosporosis.

Background: Entamoeba histolytica/Entamoeba dispar and Enterobius vermicularis are the major health problems in the developing countries especially in Iran. The prevalence of infection is variable among different social groups in the world. Objectives: Since elderly and mentally retarded are high risk group, the present survey was carried out in order to determine the prevalence of intestinal parasites especially these parasites in elderly and mentally retarded residence in Golabchi Center, Kashan, Iran. Materials and Methods: In this Cross-sectional study a total of 243 stool samples and 279 Scotch tapes from elderly and mentally retarded people were collected. Intestinal parasitic infections especially E.histolytica/E.dispar was determined by Stool examination. Scotch tape was used for diagnosis of Enterobius vermicularis. The demographic data were recorded by questionnaire and were analyzed by SPSS and X2. Results: The overall infection rate of intestinal parasite was 78.7% (191 out of 243 subjects).The prevalence of E.histolytica/E.dispar and E.vermicularis in elderly were 16.8%, 25.5% and in mentally retarded 15%, 49.1% respectively. Prevalence of pathogenic parasites was: Taenia spp. 1.6%, Hymenolepis nana 0.8%, one case of Strongyloides stercoralis, Blastocystis hominis 33.3%, Giardia lamblia 4.5%, Dientamoeba fragilis 1.6%. The rate of infection in mentally retarded was higher than elderly (P

Background: Congenital toxoplasmosis (CT) is the result of maternal infection if acquired during gestation, an infection that most often is clinically unapparent. Objectives: This study was conducted to determine acute and chronic toxoplamosis in pregnant women and its complications in newborn. Patients and Methods: In this cohort study, 798 pregnant women were tested for Toxoplasma IgG/IgM by ELISA. PCR and IgM were performed on 4 newborns of acute and 5 chronic maternal infections; they were followed for 1 year. Data were analyzed by SPSS-11.5 and χ2 test. Results: Five (0.63%) and 341 (42.7%) were positive for acute and chronic infections. A 400 bp band was seen in 3 newborns of acute maternal infections. The incidence of neonatal toxoplasmosis was 3.7 per 1000. Only one neonate of the acute maternal toxoplasmosis had jaundice. Conclusions: The rate of mate

Neospora caninum is an intracellular protozoan parasite with a wide range of intermediate bird hosts. There is little information describing the prevalence and genetic characterization of N. caninum in bird hosts worldwide and in Iran. In this study, a total of 217 brain samples of house sparrow (Passer domesticus) were examined for N. caninum presence by nested polymerase chain reaction targeting the Nc-5 gene. N. caninum DNA was detected in 3.68% (8/217) of sparrows. Sequencing of the Nc5 genomic DNA revealed 97–99% of similarity with N. caninum sequences deposited in Genbank. To our knowledge, this study is the first molecular evidence of N. caninum DNA in bird hosts in Iran. The results of this study highlight the role of the house sparrow (Passer domesticus) in maintaining and spreading N. caninum infection to canines in the feral and domestic environment.

Background: Insecticide resistance is one of the serious problems for German cockroach control program. This study was conducted to determine the bendiocarb and Carbaryl resistance mechanisms in German cockroaches using the piperonyl butoxide (PBO). Methods: Bioassay tests were conducted with 4 to 6 different concentrations of both insecticides with four replicates of 10 susceptible strain cockroaches per concentration to determine of discriminative concentration. After determining discriminative concentration, the result was compared to wild strain. The levels of susceptibility and resistance ratio (RR) and synergism ratio (SR) were calculated for each five wild strains. Moreover resistance mechanisms in wild strains were determined using PBO synergist in vivo. Results: Hospital strains showed different levels of resistance to bendiocarb and carbaryl compared to susceptible strain. The bendiocarb and carbaryl resistance ratios ranged from 2.11 to 7.97 and 1.67 to 2 at LD50 levels, respectively. The synergist PBO significantly enhanced the toxicity of bendiocarb and carbaryl to all strains with different degrees of synergist ratio, 1.31, 1.39, 3.61, 1.78, 1.62 and 2.1 fold for bendiocarb, 1.19, 1.18, 1.12. 1.29, 1.45 and 1.11-fold for carbaryl, suggesting monooxygenase involvement in bendiocarb and carbaryl resistance. Conclusion: The synergetic effect of PBO had the highest effect on bendiocarb and resistance level was significantly reduced, which indicates the important role of monoxidase enzyme in creating resistance to Bendiocarb. Piperonyl butoxide did not have a significant synergistic effect on carbaryl and did not significantly break the resistance.

Objective To determine the prevalence and economic losses of Fasciola spp. isolates and costs incurred due to liver condemnation from ruminants in Kashan region, center of Iran from 2012 to 2013. Methods In this cross-sectional study, a total number of 251 325 slaughtered sheep, goats and cattle (88 939 sheep, 151 924 goats and 10 462 cattle) were examined via necropsy for fasciolasis in industrial Kashan abattoirs located in center of Iran. The average weight was calculated as 1 000 g for sheep, 900 g for goat and 5 000 g for cattle in this study region. Based on market value, the average price of a kilo gram of normal liver at Kashan city during the study period was 8 USD. The amount of main nutrients and vitamins elimination in liver contaminated to fascioliasis was determined. Results Overall 3.28% of the livers were found to be infected. For total number of sheep, goats and cattle slaughtered annually in region study, it was estimated that 7 505 livers were infected and total annual economic losses of fascioliasis of studied animals was 41 784 USD (based on market prices in study period). Of this, 23 360 USD, 30 240 USD and 15 400 USD were associated with the fascioliasis of sheep, goats and cattle, respectively. Conclusions This study indicated that the Fasciola sp. clearly remains prevalent and causes considerable economic losses in study region in sheep, goats and cattle and presumably, other areas and hosts of Iran, providing baseline data for the future monitoring of this potentially important parasitic infection in the country.