Molecular identification of Sarcocystis species in raw hamburgers using PCR–RFLP method in Kashan, central Iran

The prevalence of bovine Sarcocystosis is high in the most regions of the world. It can be a human health problem due to consumption of raw or under cooked hamburgers or other bovine meat products. This study was carried out to investigate the prevalence and species identification of Sarcocystis among of hamburgers, using PCR–RFLP methods in Kashan, central Iran. Overall 200 raw industrial hamburgers samples with at least 60% meat were randomly collected from nine different brands in Kashan, central Iran. The genomic DNA was extracted and a PCR–RFLP method was used to amplify an approximately 900 bp fragment at the 18S rRNA(SSU) gene, restriction enzyme BclI was used for species identification. The results showed that 58 (29%) of 200 tested hamburger samples were infected to Sarcocystis spp. The prevalence rate was 31.25 and 26.9% in the hamburgers with 90 and 60–75% meat, respectively. According to PCR–RFLP analysis, 43 (74.1%) of the 58 isolates were Sarcocystis cruzi, 12 (20.7%) showed co-infection to S. cruzi and Sarcocystis hirsuta, 2 (3.5%) was mixed infected to S. cruzi and Sarcocystis hominis, 1 (1.7%) showed the pattern of mix infection to three species. This study revealed one-third of industrial hamburger were infected to S. cruzi or mixed infection of S. cruzi with other bovine sarcocytosis. To prevent cattle infection, the possible ingestion of the disposal sporocyst stage from dogs must be eliminated. Although in this study, the prevalence of S. hominis was low and cannot be considered as a major zoonosis, it should be recommended avoiding eating under cooked hamburger and other bovine meat products to prevent human infection

Molecular detection of Trichostrongylus species through PCR followed by high resolution melt analysis of ITS-2 rDNA sequences

Polymerase chain reaction followed by high resolution melting (PCR-HRM) analysis is a simple, rapid and accurate method for molecular detection of various nematode species. The objective of the present study was, for the first time, to develop a PCR-HRM assay for the detection of various animal Trichostrongylus spp. A pair of primers targeting the ITS-2 rDNA region of the Trichostrongylus spp. was designed for the development of the HRM assay. DNA samples were extracted from 30 adult worms of Trichostrongylus spp., the ITS-2-rDNA region was amplified using PCR, and the resultant products were sequenced and characterized. Afterwards, the PCR-HRM analysis was conducted to detect and discriminate Trichostrongylus spp. Molecular sequence analysis revealed that 24, 4, and 1 of the samples were T. colubriformis, T. vitrinus and T. capricola, respectively. Results from PCR-HRM indicated that complete agreement was relatively found between speciation by HRM analysis and DNA sequencing for the detection of Trichostrongylus species. The PCR-HRM analysis method developed in the present study is fast and low-cost; the method can be comparable with other molecular detection techniques, representing a reliable tool for the identification of various species within the Trichostrongylus genus. © 2020 Elsevier B.V

Genetic variation of Giardia lamblia isolates from food-handlers in Kashan, Central Iran

<p><strong><em>Background:</em></strong> Based on genotyping study of human isolates of <em>Giardia lamblia</em>; humans are mainly infected by two assemblages A and B. The present study was carried out to determine the sub-assemblages of <em>G. lamblia</em> isolated from food handlers referred to Kashan health centers, central Iran, 2015.</p><p><strong><em>Methods:</em></strong> In this cross-sectional<strong> </strong>study,<strong> </strong>3653 stool samples collected from food-handlers that annually refer to health center for getting a health certification and examined microscopically for <em>G. lamblia</em> cyst. Totally, 44 isolates were selected from 47 <em>Giardia</em> 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 <em>(gdh</em>) locus, restriction enzymes <em>BspLI </em>and <em>RsaI</em> were used for distinguish between genotypes A and B and their subgroups.</p><p><strong><em>Results:</em></strong> 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.</p><strong><em>Conclusion:</em></strong> 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 <em>Giardia</em> assemblages and sub-assemblage, more studies in human and animal population in different regions are necessary

A Dog with Multiple Infections of Enteric Parasitic Zoonosis in Mashhad City, North-East of Iran; a Case Report

Aims: In this study, we examined stool specimen from a 3-year-old domesticated dog, which was referred to a veterinary clinic with clinical signs such as nausea or vomiting, dysentery, cachexia and rash in Mashhad city, northeast of Iran. Patient &amp; Methods: A 3-year-old pet dog was referred to veterinary clinic of Mashhad in February 2016 by symptoms including, nausea or vomiting, dysentery, cachexia and rash in Mashhad City, Northeast of Iran. For parasitological examination, formalin-ether concentration technique was used. Fecal smears were made from the sediment, stained with iodine and observed by light microscope. Modified Ziehl Neelsen method was used for the detection of Cryptosporidium spp. Findings: The animal was infected with 10 disease-causing parasites; Taenia spp., Fasciola spp., Dicrocoelium dendriticum, Acanthocephal spp., Trichuris vulpis, Hook worm, Giardia spp., Blastocystis spp., Eimeria spp., and Cystoisospora spp. Conclusion: Domestic and stray dog could be an important sources for distribution of zoonoses disease especially parasitic agents

The study of relationship between serum levels of soluble fms-like tyrosine kinase-1 and soluble fibrinogen-like protein 2 with delayed graft function after kidney transplantation

Delayed graft function (DGF) is a transplant complication which means a need to dialysis throughout the first week after transplantation. This study aimed to ascertain the relationship between the two immunomodulatory factors of soluble fms-like tyrosine kinase-1 (sFlt-1) and soluble fibrinogen-like protein 2 (sFGL-2) with DGF after transplantation. This case-control study was done in 2 groups of 58 kidney transplant patients with and without DGF. The control group included the patients who didn't show DGF symptoms. Then, serum levels of sFlt-1and sFGL-2 in all blood samples were measured by ELISA. Serum sFlt-1 and sFGL-2 levels were significantly higher in the DGF group compared to those in the control group (p�0.001). sFlt-1 and sFGL-2 serum levels significantly affect DGF (p<0.001) in such a way that they may be diagnostic factors of DGF. This study showed a significant relationship between sFlt-1 as well as sFGL-2 and DGF. Therefore, plasma levels of sFlt-1 and sFGL-2 may be used as diagnostic tools to determine the risk of DGF. © August 2019, Iran J Allergy Asthma Immunol

The effect of alcoholic extracts of Arctium lappa L. and Satureja hortensis L. against Trichomonas vaginalis in vitro

Background: Trichomonas vaginalis infection is one of the most prevalent type of vaginitis in women. Considering the side effects of metronidazole and therapeutic properties of Arctium lappa L. and Satureja hortensis L. in traditional medicine, this study aimed to examine the anti-Trichomonas effects of Arctium lappa and Satureja hortensis alcoholic extracts in vitro. Materials and Methods: This experimental study was conducted on T. vaginalis isolated from 1203 persons referred to five health centers in Kashan city. Five T. vaginalis isolates were cultured in a TYI-S-33 medium and were used to study the effect of Arctium lappa and Satureja hortensis extracts. The effects of different concentrations (12.5, 25, 50, 100, 200, 400, 800 and 1000 µg/mL) of plant extracts on the growth of T. vaginalis trophozoeites were studied 12, 24, and 48 h after the culture. Also, the culture media and metronidazole (0.025, 0.05, 0.1, 0.2, 0.4 µg/mL) were considered as the negative and positive controls, respectively. The effects of the extracts and drug were examined by counting the number of live and dead parasites using the trypan blue staining method. Results: Results showed that the alcoholic extracts of Satureja hortensis and Arctium lappa had an inhibitory effect on the growth of T. vaginalis. The IC50 values of the alcoholic extracts of Satureja hortensis, Arctium lappa and metronidazole after 24 h were 190.8, 996.7 and 0.0326 µg/mL, respectively. Conclusion: The present study shows the in vitro anti-Trichomonas effect of Arctium lappa and Satureja hortensis extracts. The anti-Trichomonas effect of Satureja hortensis extract was higher than that of the Arctium lappa extract. Further studies are recommended to investigate the anti-Trichomonas effect of major components of these plants, especially the Satureja hortensis extract

Induction of apoptosis by alcoholic extract of combination verbascum thapsus and ginger officinale on Iranian isolate of trichomonas vaginalis

Introduction: The genus Malassezia is an important skin resident of human. The present study aimed to analyze in vitro activity of the essential oils of Lavandula stoechas, Cuminum cyminum and Artemisia sieberi against clinical strains of Malassezia species. Methods: A total of 47 Malassezia strains, including Malassezia furfur, Malassezia globosa and Malassezia obtuse, were used in this study. A disk diffusion technique was selected for testing the susceptibility of Malassezia strains to the essential oils. Results: All the essential oils showed in vitro activity against Malassezia strains, with M. furfur and M. obtusa being the highest and lowest susceptible of the strains, respectively. The highest antifungal activity was associated with the essential oil of C. cyminum (mean ± SD: 50.0 ± 0.0 mm), followed by L. stoechas (mean ± SD: 46.8 ± 3.1 mm) and A. sieberi (mean ± SD: 36.9 ± 5.7 mm). The inhibition zone ranges were 12.5 to 15.6 mm (mean ± SD: 14.4 ± 1.6 mm) for ketoconazole and 11.6 to 13.3 mm (mean ± SD: 12.4 ± 0.9 mm) for fluconazole. Although all the antifungal drugs were found to possess good antifungal activities against Malassezia strains, their effects were lower than the activities shown by the essential oils tested (P < 0.05). Conclusion: These results indicated that the essential oils tested, especially the one from C. cyminum, inhibited the growth of clinical strains of Malassezia, implying its potential use in the treatment of Malassezia infections. This indicates that this plant may be useful in preparation of new drugs

Local and non-local equivalent potentials for p-12C scattering

A Newton-Sabatier fixed energy inversion scheme has been used to equate inherently non-local p-${}^{12}$C potentials at a variety of energies to pion threshold, with exactly phase equivalent local ones. Those energy dependent local potentials then have been recast in the form of non-local Frahn-Lemmer interactions.Comment: 15 pages plus 9 figures submitted to Phys. Rev.

Seroepidemiology of Toxoplasma gondii infection in immunodeficiency patients in Kashan and Qom during 2014-2015

Background: Toxoplasma gondii is an opportunistic parasitic protozoon, which is a causative agent of serious complications such as encephalitis in immunodeficiency patients. Considering insufficient data on toxoplasmosis in these patients, the present study was conducted to determine the seroepidemiology of T. gondii among immunodeficiency patients. Materials and Methods: This cross-sectional study was conducted on cancer, ADIS, hemodialysis and renal transplant patients (case group) and healthy persons (control group) in Kashan and Qom cities. Totally, 317 serum samples were collected from the case group and 120 samples from the control group. The ELISA method was used to determine the anti-T. gondii IgG and IgM antibodies. Results: Totally, 60.3 of the samples from the case and 33.3 from the control groups were positive for anti-T. gondii IgG (P&lt;0.001). In the case group, only 2 persons (0.6) were positive for anti-T. gondii IgM. The anti-T. gondii IgG detected in immunodeficiency patients was 55.2 in Kashan and 68 in Qom, which were higher than in the control group (P&lt;0.001, P&lt;0.006, respectively). The highest prevalence of T. gondii were seen in 40-59 years old (49.8) and illiterate (60) patients. There was a meaningful correlation between toxoplasmosis and blurry vision and dermal rash (P=0.001 and P=0.003, respectively). Conclusion: The prevalence of T. gondii was higher in different immunodeficiency patients compared to healthy persons. Screening examinations were recommended for the diagnosis and treatment of patients to prevent serious side-effects and health education

Detection of Drug Resistance Gene in Cutaneous Leishmaniasis by PCR in Some Endemic Areas of Iran

Background: Cutaneous leishmaniasis is still a health problem in many rural and urban regions of Iran and drug resistance has emerged as a major impediment in the treatment of leishmaniasis. This study aims to determine the drug resistance gene in cutaneous leishmaniasis by PCR in some endemic areas of Iran. Methods: Ninety seven samples were collected from ulcers of leishmaniasis patients from some endemic areas of Iran. The Giemsa stained samples were examined microscopically and cultured in NNN and RPMI 1640 mediums for parasite detection. After DNA extraction, PCR was done by a pair of specific primers. For detection of mutation in DNA, first PCR products were electrophoresed on CSGE gel. The suspected samples were compared by sequencing and RFLP results were demonstrated. Comparison of DNA derived from a wild type cell and mutant cell was undertaken by CSGE and sequencing methods. Results: Among 90 isolates (92.8) examined for detection of mutation in gene with CSGE and RFLP, 10 (11.1) revealed a disorder in sequencing selection for unresponsive to drug. Conclusion: Drug resistance in cutaneous leishmaniasis to sodium stiboglocanat is probably due to a mutation in a genome. A field study is needed to determine the distribution of drug resistance and other gene mutations involved in unresponsiveness to drugs in leishmaniasis endemic areas of Iran. © Iranian Red Crescent Medical Journal