Molecular epidemiology of Blastocystis spp. in children referred to Qods hospital in northwest of Iran.

Blastocystis sp. is a polymorphic intestinal parasite in humans and animals. The parasite has a worldwide distribution, especially in developing countries with poor sanitation, exposure to animals, and improper disposal systems. The aim of this study was to identify the subtypes of Blastocystis sp. among children of Qazvin, northwest Iran. Totally, 864 stool samples were collected from the children referred to Qods hospital in Qazvin, Iran. Fecal specimens were investigated by formalin-ethyl acetate concentration method and trichrome staining as well as cultivation of all samples in clotted fetal bovine medium. DNA extraction of culture-positive specimens and PCR amplification of 18S ribosomal RNA gene region was performed. The sequences detected were compared with reference genes in the GenBank, and the sequences further deposited in the GenBank database. Data analysis was performed by Chi square test while a p value of < 0.05 was considered as significant. Of 864 isolates, 4.1% (36/864) were positive for Blastocystis sp. with infection rate insignificantly higher among the females than males. The highest infection rate was estimated at 6.8% in 6-9 years old age group with abdominal pain as the most common (33%) gastrointestinal sign. No statistically significant difference was found between the variables and Blastocystis infection. Molecular analysis clarified the presence of three subtypes of Blastocystis including ST1 (56%), ST2 (28%), and ST3 (16%) of among specimens with ST1 as the predominant subtype. A significant association between intestinal signs and the subtypes was not found. Considering ST1 as the predominant subtype, it seems that zoonotic transmission is a main route of human infections with Blastocystis sp. in the area studied.

Distribution and molecular analysis of Blastocystis subtypes from gastrointestinal symptomatic and asymptomatic patients in Iran.

INTRODUCTION: Blastocystis is a common intestinal parasite of human and animal hosts. The parasite has 17 subtypes, and among those at least nine subtypes (ST1-ST9) are found in human hosts. OBJECTIVE: The aim of the present study was to investigate the presence of different subtypes of Blastocystis spp. among the patients referred to Velayat hospital of Qazvin province, Iran. METHODS: Overall, 864 stool samples were examined by using formalin-ethyl acetate concentration method and Trichrome staining. All specimens were cultured in clotted fetal bovine medium. Later, DNA extraction and PCR amplification of 18S ribosomal RNA gene region was conducted and phylogenetic tree constructed. RESULTS: The results revealed 7.9% (68/864) of the study population were infected with Blastocystis. Intestinal symptoms were observed in 61% (36/59) of individuals positive for Blastocystis, with abdominal pain in 58% (21/36) of cases which was more frequent than other intestinal signs. No significant relationship was observed among the study variables. By molecular and phylogenetic analysis, three subtypes ST1 (45%), ST2 (30%) and ST3 (23%) of parasite were identified. CONCLUSION: This study showed ST1 subtype was the predominant subtype among the positive specimens, meanwhile the highest haplotype and nucleotide diversity were clarified in ST3 subtype.

Quality of Life in Different Chronic Diseases and Its Related Factors.

BACKGROUND: Quality of life (QoL) is an important tool for evaluating the effect of a disease as well as effects of treatment interventions. The present study is aimed to investigate QoL of the patients with various prevalent chronic diseases in Iran. METHODS: In a cross-sectional study, 625 patients with one of the prevalent chronic diseases were assessed. Participants were recruited via convenient sampling method in special clinics of chronic diseases in three educational hospitals in Qazvin, Iran, from May to December 2016. Data were collected using WHOQOL-BREF and demographic questionnaire through interview. Data were analyzed using independent t-test, ANOVA, Pearson's correlation coefficient, and multiple linear regression method. RESULTS: Significant difference in QoL scores was found in different chronic disease; patients with diabetes and hypertension had higher scores and patients with asthma and chronic obstructive pulmonary disease (COPD) had the lowest scores. Multivariate analysis showed that age, economic status, and type of disease were among the significant variables in predicting the QoL of the patients with chronic diseases. CONCLUSIONS: Some patients, such as those with asthma and COPD, had less QoL than other patients and should be prioritized while planning for the promotion of healthcare services. Due to the negative effects of economic status on QoL, the financial support programs should be among the major priorities of the patients' QoL improvement programs.

Molecular characterization of Hymenolepis nana based on nuclear rDNA ITS2 gene marker.

INTRODUCTION: Hymenolepis nana is a zoonotic tapeworm with widespread distribution. The goal of the present study was to identify the parasite in the specimens collected from NorthWestern regions of Iran using PCR-sequencing method. METHODS: A total of 1521 stool samples were collected from the study individuals. Initially, the identification of hymenolepis nana was confirmed by parasitological method including direct wet-mount and formalin-ethyl acetate concentration methods. Afterward, PCR-sequencing analysis of ribosomal ITS2 fragment was targeted to investigate the molecular identification of the parasite. RESULTS: Overall, 0.65% (10/1521) of the isolates were contaminated with H. nana in formalin-ethyl acetate concentration. All ten isolates were succefully amplified by PCR and further sequenced. The determined sequences were deposited in GenBank under the accession numbers MH337810 -MH337819. CONCLUSION: Our results clarified the presence of H. nana among the patients in the study areas. In addition, the molecular technique could be accessible when the human eggs are the only sources available to identify and diagnose the parasite.

Prevalence of Parasitic Intestinal Infections Among Food Handlers in Qazvin, Iran

Objective: Food handlers play an important role in the production and distribution of parasitic contaminations in these societies. The aim of this study was to assess the prevalence of intestinal parasitic infections among the food handlers in Qazvin, Iran. Methods: Totally, 1530 stool samples were collected from food handlers in Qazvin, Iran. Direct feces examination and formalinethyl acetate concentration method followed by Trichrome staining were performed to detect the parasites. Results: The prevalence of intestinal parasites was 82 (5.4%) among food handlers. The species of protozoan parasites found were Blastocystis hominis, Giardia lamblia, Entamoeba coli, and Endolimax nana. The outbreaks of Blastocystis hominis (2.9%), and Endolimax nana (0.3%) were the most and the least protozoan infections observed among the specimens, respectively. Also, there was no helminthic infection detected among the samples. The prevalence of intestinal parasites among the food handlers, based on variables including sex, age, job, and education, did not differ, whereas the prevalence of intestinal symptoms was shown to be higher in infected food handlers (p<0.05). Conclusions: Despite a decrease in the incidence of intestinal parasites, the protozoan infections are still among the health concerns in these areas. Therefore, continuous monitoring and training of food handlers by local health authorities should be performed on a regular basis.

Prevalence of intestinal parasites in food handlers of the city of Saqqez in 2016.

This study was aimed at determining the prevalence of parasitic infections among the food handlers of Saqqez County in 2016. Stool samples collected from 1530 food handlers. All samples were examined by both direct technique and sedimentation technique using formalin-ethyl acetate for detecting the intestinal parasites. Of total participants, 1462 (95.6%) were male and 62 (4.4%) female. Intestinal parasitic infections were found in 92 (6.01%) individuals. The highest infection rate was due to Giardia lamblia (4.1%) and the lowest associated with Endolimax nana, Iodamoeba butschlii, and Hymenolepis nana (0.1%). No significant correlation between the intestinal parasitic infections and age, gender, education level, gastrointestinal signs and symptoms, and the type of profession, was established. The results of the present study revealed that only a small number of food handlers in Saqqez County were infected with intestinal parasites with G. lamblia as the most common cause of infection. More strict health control for those working in food industries is of vital importance and continuous education to improve the level of general health in these people is undoubtedly considered as a key determinant towards lower number of parasitic infections.

Investigation of Giardia intestinalis Genotypes among the Food Handlers of Qazvin, Iran.

BACKGROUND: We aimed to investigate the genotypes of Giardia intestinalis among the food handlers in Qazvin, Iran. METHODS: Overall, 1530 stool specimens were collected from the food handlers who visited Shahid Bolandian Health Center, Qazvin, Iran during 2016. Specimens were evaluated by microscopic and concentration methods. Twenty specimens with appropriate number of giardia cysts were selected followed by DNA extraction. Determination of giardia genotypes was achieved through PCR and sequencing the glutamate dehydrogenase gene. The phylogenetic tree was drawn using the MEGA7 software. Finally, the data were analyzed statistically with a P-value<0.05 was considered as significant. RESULTS: Twenty stool samples (1.3%) were positive for Giardia cyst. All positive specimens were obtained from male participants with abdominal cramp being their most common symptoms. The mean age for infected individuals was 32 yr. Molecular characterization was successfully performed for 17 isolates and two genotypes A (AII, 65%) and B (BIII, 35%) were identified. CONCLUSION: The most prevalent giardia genotypes among the food handlers in Qazvin were A (AII) and B (BIII) genotypes with A (AII) genotype as the dominant one in the region. Considering the direct association between the food handlers and public health as well as the impact of geographical and host conditions on dispersion and pathogenicity of various genotypes and their zoonotic aspects, further investigations are necessary.

Evaluating the effect of Myrtus communis on programmed cell death in hydatid cyst protoscolices.

OBJECTIVE: To evaluate the possible involvement of programmed cell death strategy in hydatid cyst protoscolices following treatment with Myrtus communis (M. communis) as an herbal medicine. METHODS: Protoscolices were aseptically collected from sheep liver hydatid cysts. Evaluating the effect of M. communis extract on programmed cell death and increased activity of caspases 3, 8, and 9 in hydatid cyst protoscolices was conducted by treating the protoscolices with different concentration (5, 50, and 100 mg/mL) of M. communis extract at 37 °C and 5% CO2 for 4 h by using the Bradford test and ELISA commercial kits. RESULTS: The extract of M. communis at all concentrations led to initiation of programmed cell death in protoscolices and this effect, was only significant at 50 and 100 mg/mL concentrations, compared to the negative control (P < 0.05). Also, the activity of caspases 3, 8, and 9 in hydatid cyst protoscolices, was shown that the extract at all 3 concentrations could only increase the activity of caspases 3 and 9. Moreover, a significant increase in the activity of caspase 3 was only observed at concentrations 50 and 100 mg/mL by 37.00% and 66.19% while a significant increase in the activity of caspase 9 at the same concentrations was observed by 20.89% and 63.67%, respectively (P < 0.05). CONCLUSIONS: The extract of M. communis at different concentrations could increase the activity of caspases 3 and 9 and caused programmed cell death in hydatid cyst protoscolices however, this effect was significant at high concentrations of the extract.

Protoscolicidal and immunomodulatory activity of Ziziphora tenuior extract and its fractions.

OBJECTIVE: To evaluate the scolicidal and immunomodulatory effect of the Ziziphora tenuior (Z. tenuior) extract and its fractions. METHODS: Protoscolices were treated with six concentrations (3, 5, 10, 25, 50, and 100 mg/mL) of Z. tenuior extract and its fractions (ethanol, petroleum ether, ethyl acetate and chloroform) in periods of 10, 20, 30, 40, 50 and 60 min, and viability of protoscolices was evaluated using the 1.0% eosin. To examine the immunomodulatory effects of Ziziphora and its fractions on macrophage cells, the non-toxic concentration of extract and different fractions determined by MTT assay, and the Griess reaction was used to measure the level of nitrite as an indicator of nitric oxide by the macrophage cells in 10, 100 and 200 µg/mL in 24 h at 37 °C. RESULTS: In this study, the Z. tenuior extract at 10 mg/mL concentration was able to kill all protoscolices during 20 min. By increasing the concentration to 25 mg/mL, the scolicidal time reduced to 10 min. Regarding the effect of different fractions of Z. tenuior, the ethanolic fraction showed the highest scolicidal activity. The extract demonstrated an inhibitory effect on the activity of macrophages and reduced nitric oxide production. Although the petroleum ether and ethanolic fractions of the extract reduced nitric oxide production, nevertheless, this effect was only significant at 10 and 100 µg/mL concentrations (P < 0.05). CONCLUSION: The Z. tenuior extract and its fractions were effective against protoscolices yet the effect of total extract was considerable. Our findings indicates that the extract and its ethanolic and petroleum ether fractions could have anti-inflammatory properties.

In vitro protoscolicidal effects of various concentrations of Ziziphora tenuior L. extract at different exposure times.

OBJECTIVE: Hydatidosis is one of the most important zoonotic diseases and surgery is still the main treatment for this problem. One of the side effects of hydatid cyst surgery is recurrence, thus, searching and assessment of some new agents such as medicinal plant extracts are very important. In the present study, the scolicidal effect of ethanolic extract of Ziziphora tenuior (Z. tenuior) was investigated. MATERIALS AND METHODS: Protoscolices were aseptically collected from sheep livers containing hydatid cyst and used in the experiments. Z. tenuior extract was used at concentration of 3-100 mg/ml for 10-60 min. Viability of protoscolices was determined by 0.1% eosin staining. RESULTS: Based on our results, Z. tenuior extract at concentration of 10 mg/ml killed all protoscolices after 20 min. However, this medicinal plant at concentration of 25 mg/ml destroyed all protoscolices in a shorter exposure time (10 min). Therefore, the scolicidal activity of the extract at 10 and 25 mg/ml concentrations was considerably effective in lower concentrations and shorter exposure times. CONCLUSION: The findings of this study showed that the ethanolic extract of Z. tenuior produces high scolicidal activity; it may be used as an appropriate and effective scolicidal agent in hydatidosis surgery. This is the first report on the protoscolicidal activity of Z. tenuior.

Study of Protoscolicidal Effects of Hypertonic Glucose on Protoscolices of Hydatid Cyst at Different Concentrations and Exposure Times.

Surgical operation is the treatment of choice for hydatid cyst. To date, many protoscolicidal substances have been used for inactivation of hydatid cyst contents but most of these compounds may produce several side effects. The aim of this study was to evaluate the protoscolicidal effects of hypertonic glucose on protoscolices of hydatid cyst at different concentrations and exposure times. Protoscolices were obtained aseptically from the livers of slaughtered sheep at Qazvin abattoir, Iran. Protoscolices were exposed to different concentrations of hypertonic glucose (10%, 15%, 20%, 25%, 30%, 40%, and 50%) at different times (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, and 60 min). Viability of protoscolices was evaluated by 0.1% eosin and the movement of protoscolices flame cells. The highest protoscolicidal effect (100%) of hypertonic glucose was obtained at concentrations 40% and 50% following 40 and 20 min exposure times, respectively. Some protoscolicidal agents show a variety of dangerous complications such as biliary tract fibrosis and liver necrosis; however, hypertonic glucose especially at a concentration of 40% may demonstrate less side effects compared with glucose 50%. Further in vivo investigations are suggested.