Identification and multilocus genotyping of Toxoplasma gondii isolates from congenital infection in north of Iran.

Congenital toxoplasmosis can cause severe consequences in the fetus, such as spontaneous abortion which is affected by parasite strain. Also, recent studies revealed the high genetic diversity of Toxoplasma gondii. This study aims to investigate the serological status of T. gondii in pregnant women, multilocus genotyping in aborted fetuses' tissue, and archived formalin-fixed paraffin-embedded placenta. This study was performed on 100 pregnant women with spontaneous abortion and their aborted fetuses, and 250 of the archived placentae in Iran. The blood and tissue were examined for seroprevalence and genotype determination of T. gondii using ELISA and multilocus nested-PCR-RFLP, respectively. Anti-T. gondii IgG and IgM were detected in 68 samples (68%) and 1 (1%) out of 100 serums. Toxoplasma DNA was identified in 1 (1%) aborted fetuses' tissue and 32 (12.8%) placenta samples. Overall, ten positive DNA samples were successfully genotyped, and five genotypes were recognized (ToxoDB#1, #2, #10, #27, and #48). The obtained results indicated congenital toxoplasmosis is a severe risk in this region. As type I is highly pathogen and can lead to severe complications, the prevention of the infection should be considered in seronegative pregnant women.

Genetic characterization of Toxoplasma gondii in meat-producing animals in Iran.

BACKGROUND: The consumption of uncooked or undercooked food from infected intermediate hosts can result in Toxoplasma gondii infection in humans. However, few studies have investigated the genetic diversity of this protozoan parasite in Iran. The aim of the present study was to genetically characterize isolates of T. gondii from intermediate host animals in Mazandaran Province, Iran. METHODS: Blood and heart tissue samples were collected from 204 ruminants, and brain tissue was collected from 335 birds. The prevalence of T. gondii infection in these samples was determined serologically using the modified agglutination test and by conventional PCR assays. Those PCR samples positive for T. gondii DNA and 13 DNA samples extracted from aborted fetuses in a previous study were genotyped with 12 genetic markers using the multilocus-nested PCR-restriction fragment length polymorphism (Mn-PCR-RFLP) technique. RESULTS: Antibodies for parasites were found in 35.7% of the ruminant (39.1% of sheep and 26.4% of goats) samples and in 51.3% of the bird (100% of geese, 52.3% of free-range chickens and 46% of ducks) samples. Molecular detection by PCR of the repetitive 529-bp DNA fragment revealed contamination of 13.2% of ruminant (14.6% of sheep and 9.4% of goats) samples and of 9.6% of bird (11.1% of chickens, 5.7% of ducks and 0% of geese samples). The results from 30 DNA samples revealed five ToxoDB (genome database for the genus Toxoplasma) PCR-RFLP genotypes, including #1 (Type II), #2 (Type III), #10 (Type I), #27 and #48, with genotype #1 the most predominant. CONCLUSIONS: As evidenced by the results of this study, ruminants and birds are infected with T. gondii in the region, suggesting that they are likely to be involved in the transmission of T. gondii to humans through meat consumption. The identification of different genotypes may suggest a higher genetic diversity of this parasite in Mazandaran, reflecting local environmental contamination. These results have important public health implications for the prevention and control strategies of infection.

Toxoplasmosis among cancer patients undergoing chemotherapy: a population study based on the serological, molecular and epidemiological aspects.

BACKGROUND: Toxoplasmosis is highly prevalent in northern Iran and immunocompromised individuals are more susceptible to this infection. The present study aimed to determine the seroprevalence, parasitism and genetic diversity of Toxoplasma gondii among patients with cancer undergoing chemotherapy in northern Iran. METHODS: A total of 350 serum samples obtained from cancer patients were collected from laboratory centers in northern Iran. Immunodiagnosis and DNA detection were accomplished by ELISA and PCR. Thereafter, multiplex-nested PCR-restriction fragment length polymorphism was used for the genotyping of T. gondii. RESULTS: In general, out of 350 patients, 264 (75.4%) and 9 (2.57%) cases were positive for anti-T. gondii IgG and IgM, respectively. Moreover, 19 (5.43%) samples contained T. gondii DNA. From 19 positive samples, 10 high-quality samples with sharp and non-smear bands were selected to determine the genotypes of T. gondii. Accordingly, the samples were classified as genotype #1 (type II clonal; n=4, 40%), genotype #2 (type III clonal; n=3, 30%), genotype #10 (type I clonal; n=2, 20%) and genotype #27 (type I variant; n=1, 10%). CONCLUSIONS: As evidenced by the results, due to the high prevalence of T. gondii, cancer patients in northern Iran are at serious risk of severe toxoplasmosis and its complications. Therefore, oncologists need to regard this critical health problem as a matter requiring urgent attention.

The Predictors of Postoperative Pain Among Children Based on the Theory of Unpleasant Symptoms: A Descriptive-Correlational Study.

PURPOSE: Postoperative pain (POP) is a common outcome of surgical interventions among children. Identifying POP contributing factors can help identify children who are at risk for POP and facilitate POP management. The aim of the study was to determine the predictors of POP based on the Theory of Unpleasant Symptoms. DESIGN AND METHODS: This descriptive-correlational study was conducted in 2018-2019 on 153 children purposively recruited. Data were collected using a personal characteristics questionnaire, the Coping Strategies Questionnaire, the Child Pain Anxiety Symptoms Scale, and a Visual Analogue Scale for Pain. The SPSS software (v. 21.0) was used for data analysis. RESULTS: Twelve participants were excluded and final data analysis was done on the data obtained from 141 participants. Most of participants were male (65.2%), underwent abdominal surgery (53.2%), and experienced POP (86.5%). Their age mean was 8.58±2.23. Linear regression analysis revealed that the physiological factors of heart rate, preoperative pain, and surgery duration as well as the psychological factors of anxiety and coping strategies were significant predictors of POP among children. All these factors collectively explained 34% of the total variance of POP. Situational factors (such as age, gender, and ethnicity) had no significant effects on POP. CONCLUSIONS: Heart rate, preoperative pain, surgery duration, anxiety and coping as predictive factors of POP can be considered when designing effective POP management strategies. PRACTICE IMPLICATIONS: The findings provide a better understanding about the predictors of POP and can be used to develop pain management among children.

The effect of poetry reading on self-concept of primary adolescents.

OBJECTIVES: this study, regarding the importance and significance of self-concept in children's psychological health and considering the strong role of art in meeting psychological needs, aimed at investigating the effect of poetry reading on the self-concept of Adolescents. METHODS: This study was a classic experimental design of pre-test and posttest with two groups of intervention and control in the academic year 2016-2017. The research population included Adolescents who were studying in the city of Aq Qaladar in the province of Golestan, Iran. Sixty students. Data collecting tools included two questionnaires and "Piers-Harris Children's Self-Concept" scale. The experimental groups were provided with poetry reading in eight group sessions of 60 min for 4 weeks. The poems were approved by 10 teachers with 10 years of experience, and the supervisor. Finally, paired t-test and independent t-test were used to analyze data using SPSS 16 (SPSS Statistics is a software package used for interactive, or batched, statistical analysis). RESULTS: Adolescents self-concept in the control group was 57.9(4.33) and in the experimental group 62.4(4.28) out of 80 questionnaires which showed a significant difference between the two groups (p < 0.001). Which demonstrated the effectiveness of poetry reading on Adolescents self-concept. CONCLUSIONS: Considering the positive impact of poetry reading on adolescent self-esteem, this intervention is recommended to improve adolescent self-esteem.

A serological investigation and genotyping of Toxoplasma gondii among Iranian blood donors indicates threat to health of blood recipients.

BACKGROUND: Toxoplasmosis is a zoonotic disease in animals and human caused by the intracellular obligatory protozoan named Toxoplasma gondii. The purpose of this study was to evaluate the sero-molecular prevalence and genotyping T. gondii among healthy blood donors in north of Iran. METHODS: In this cross-sectional study, 400 blood donors participated from all Blood Transfusion Organization (BTO) in Mazandaran province during October and November 2014. The blood samples were investigated for seroprevalence, DNA detection and genotyping of T. gondii using ELISA, nested-PCR, and Multilocus nested-PCR-RFLP methods respectively. RESULTS: Among all of blood donors, 294 (73.5 %) and 9 (2.2 %) cases were seropositive for anti-T. gondii IgG and IgM antibodies. T. gondii DNA was detected in 7 samples. Four genotype of T. gondii were identified in blood donors samples (Genotype ToxoDB#1, #2, #10 and #27), which 50 % of T. gondii strains were highly pathogenic. CONCLUSIONS: Taking into account survive T. gondii in blood transfusion bag, the high prevalence of T. gondii and existence of pathogenic genotypes in Iranian blood donors, it seems that T. gondii screening should be performed at the BTO to prevent complications of toxoplasmosis in blood recipients.

Genetic characterization of Toxoplasma gondii in Iranian HIV positive patients using multilocus nested-PCR-RFLP method.

The aim of the present study was to investigate the prevalence and genotyping of Toxoplasma gondii in Iranian human immunodeficiency virus (HIV)-positive patients using multilocus-nested polymerase chain reaction restriction fragment length polymorphism (Mn-PCR-RFLP). A total of 102 serum samples obtained from infected patients were collected from the laboratory centres in northern Iran. Anti-T. gondii antibodies and deoxyribonucleic acid (DNA) detection were accomplished by an enzyme-linked immunosorbent assay and PCR. The Mn-PCR-RFLP method was used for the genotyping of T. gondii. Overall, 68.6% (70/102) and 11.7% (12/102) of the individuals were tested positive for anti-T. gondii immunoglobulin G and T. gondii DNA, respectively. Complete genotyping was performed on 10/12 (83.3%) PCR-positive samples. Accordingly, the samples were classified as genotype #1 (type II clonal; n = 3, 30%), genotype #2 (type III clonal; n = 2, 20%), genotype #10 (type I clonal; n = 2, 20%), genotype #27 (type I variant; n = 1, 10%), genotype #35 (type I variant; n = 1, 10%) and genotype #48 (type III variant; n = 1, 10%). The results were indicative of the high frequency of the type I and type I variant of T. gondii strains in HIV-positive patients in northern Iran. Given the high prevalence of T. gondii and frequency of pathogenic types (pathogen in laboratory mice) in the patients, special measures should be taken to prevent the possible increased incidence of encephalitis by T. gondii.

Frequency and genetic diversity of Blastocystis subtypes among patients attending to health centers in Mazandaran, northern Iran.

Blastocystis hominis is a common intestinal protozoan with global distribution. The prevalence of B. hominis is extremely diverse in different countries. Molecular studies show that there is a great deal of genetic variation in the Blastocystis parasite isolated from humans and animals. Therefore, this study was conducted to determine B. hominis genotypes in patients attending to health centers of Sari, Mazandaran Province, north of Iran. 420 fecal specimens were collected from individuals in health centers of Sari from April to December 2017. The samples were examined by direct methods, parasite test (formalin ether) and Giemsa staining. In order to determine the genotypes of Blastocystis parasite, all positive samples in direct methods were tested by PCR using SSU rDNA gene. Eventually, seven positive molecular samples were sequenced and recorded in the gene bank. The phylogenetic analysis was done by drawing a gene tree using the neighbor-joining procedure. B. hominis was detected in 60 stool specimens by microscopic method. All these 60 samples were positive by PCR and the bands 310 bp were observed. According to the phylogenetic tree, it was revealed that B. hominis isolates of Mazandaran Province are more similar to subtype ST3 (ST type) that reported in Khorramabad in Lorestan Province (West of Iran). The present study showed that, like most studies, ST3 is the dominant genotype of protozoan in Sari. Improvement of public health awareness, relative improvement in environmental health, urban wastewater treatment, proper clothing coverage and the use of gloves during dealing with livestock and the use of domestic water treatment devices can reduce the Blastocystis infection in this area.

Global status of synchronizing Leishmania RNA virus in Leishmania parasites: A systematic review with meta-analysis.

Leishmaniasis is one of the most neglected tropical diseases caused by protozoan parasites belonging to the genus Leishmania. There is much evidence regarding prevalence of Leishmania RNAvirus (LRV) causing Old World leishmaniasis (OWL) and New World leishmaniasis (NWL); however, a combined evidence-based knowledge on this topic is not still available. The purpose of this systematic review and meta-analysis was to address the global status of synchronizing LRV in Leishmania in the available literature. The data were systematically collected from the English electronic databases up to May 2018. Then, the studies were screened based on the inclusion and exclusion criteria. The random-effect model was used by forest plot with 95% confidence interval (CI). Overall, 877 samples from 17 articles were included in this study. Given species of Leishmania, the highest prevalence of LRV belonged to Leishmania (L.) Viannia (V.) guyanensis and L. V. braziliensis. Additionally, the virus was detected also in L. V. amazonensis, L. V. panamanensis, L. V. lainsoni, L. aethiopica, L. major and L. infantum. By random-effect model, the global prevalence of LRV was estimated to be 26.2% (95% CI: 14.4% - 40.1%). The high prevalence of LRV among causative agents of NWLisolated from the metastatic clinical forms suggests potential association of LRV with metastatic clinical forms in New World endemic regions. A comprehensive investigation on experimental and clinical aspects of LRV is needed to fully appraise the role of these viruses in pathogenicity of Leishmania parasites and their drug resistance.

Molecular Genotyping of the Human Cystic Echinococcosis in Mazandaran Province, North of Iran.

BACKGROUND: The larval stage of the tapeworm (cestode) Echinococcus granulosus is the etiological agent of hydatidosis or cystic echinococcosis, which is the zoonotic parasitic disease causing morbidity and mortality in both humans and livestock. Due to a lack of accurate data on the human isolates of E. granulosus in Mazandaran Province, northern Iran, the current study aimed to survey the population genetic pattern of cystic echinococcosis isolated from humans by sequencing the mitochondrial genes of NADH dehydrogenase subunit 1 (nad1). METHODS: Overall, 47 formalin fixed paraffin-embedded tissue (FFPT) blocks were collected from patients' files in various pathology departments of Mazandaran Province in Iran from 2003 to 2015. PCR was performed to amplify a 398bp DNA fragment of mitochondrial nad1. PCR products were sequenced by Bioneer Corporation (South Korea), and the resulting data were analyzed via relevant software to determine the genotypes. RESULTS: The nad1 gene was successfully amplified on 10 from all of the E. granulosus isolates. Overall, 66.6% and 33.3% of the isolates in the studied area displayed the G1 and G2-G3 genotypes, respectively. CONCLUSION: This study may provide the foundation for further studies in revealing the regional transmission patterns and also in designing adequate control procedures.

Aetiology of livestock fetal mortality in Mazandaran province, Iran.

In the farming industry, the productivity of livestock herds depends on the fertility efficiency of animals. The accurate diagnosis of a broad range of aetiological agents causing fetal death is often difficult. Our aim was to assess the prevalence rates of Toxoplasma gondii, Neospora caninum, and Brucella spp. infections in ruminant abortion using bacteriological culture and molecular techniques in Mazandaran Province, northern Iran. Samples were collected from 70 aborted sheep, goat, and cattle fetuses between September 2014 and December 2015. Necropsy was performed on all the received samples, and brain tissue and abomasal content were obtained from the aborted fetuses. Protozoan infections were detected by specific polymerase chain reaction (PCR) and bacterial agents using bacteriological examinations and PCR assay. Infectious pathogens were detected in 22 out of 70 (31.4%) examined fetuses. Moreover, T. gondii, N. caninum, and B. melitensis were verified in 13 (18.6%), four (5.7%), and two (2.85%) samples, respectively. Our results showed that infection with the mentioned pathogenic agents may lead to fetal mortality, which can be a major cause of economic loss. The listed pathogens could be considered important etiological agents of fetal loss in Mazandaran Province, for which appropriate control measures such as vaccination and biosecurity can be implemented to prevent infection and reduce reproductive loss in livestock farms.

Novel Point Mutations in cyp51A and cyp51B Genes Associated with Itraconazole and Posaconazole Resistance in Aspergillus clavatus Isolates.

Aspergillus clavatus is a common environmental species known to cause occupational allergic disease in grain handlers. We have recently observed azole-resistant isolates of this fungus as a cause of onychomycosis. To further characterize the cause of resistance, the genes encoding 14 α-sterol demethylase enzyme (cyp51A and cyp51B) were characterized and analyzed in 9 ITC-susceptible isolates and 6 isolates with high minimum inhibitory concentrations (MICs) of clinical (nail and sputum) and environmental A. clavatus strains. We found that six isolates with itraconazole MIC >16 mg/L demonstrated nonsynonymous mutations, including V51I, L378P, E483K, and E506G, and synonymous mutations, including F53F, A186A, Q276Q, and H359H. Moreover, P486S was detected in five strains with ITR MIC >16 mg/L. One mutation, F324S, was detected in an isolate with posaconazole MIC >16 mg/L. The effect of E483K and P486S mutations of CYP51A on azole resistance was further investigated using homology modeling and molecular dynamics. We found that E483K and P486S mutations were located near the ligand access channel of CYP51A that could partly lead to narrowing the entry of the ligand access channels. Therefore, we concluded that E483K and P486S mutations may potentially contribute to the limited access of inhibitors to the binding pocket and therefore confer resistance to azole agents.

A multiplex restriction enzyme-PCR for unequivocal identification and differentiation of Trichostrongylus species in human samples.

Trichostrongylus species remain one of the major health challenges in the tropical and summer rainfall regions worldwide. Identification of strongylid species diagnostic methods is vital for obtaining a deep understanding of the epidemiology, population biology, anthelmintic treatment efficacy, and drug resistance in order to design effective parasite control strategies. We evaluated a multiplex RE-PCR for the diagnosis of key Trichostrongylus spp. Genomic DNA amplification of Trichostrongylus colubriformis, Trichostrongylus axei and Trichostrongylus vitrinus was achieved as standard sample using specific primers located in the second internal transcribed spacer (ITSII) of nuclear ribosomal DNA (rDNA). The mentioned method was based on isolation of Trichostrongylus ova from human fecal samples using Willis method, the extraction of ova genomic DNA samples, followed by rDNA ITSII PCR and one-step multiplex RE-PCR using three restriction enzymes of HinfI, DraI, and MseI. The multiplex RE-PCR technique provides a useful tool for discriminating all Trichostrongylus spp., being useful for diagnostic, epidemiological, ecological studies, and control programs. This method is rapid, especially when numerous restriction enzymes are required for species differentiation or identification.

Intestinal parasitic infections in Iranian preschool and school children: A systematic review and meta-analysis.

Parasitic infections are a serious public health problem because they cause anemia, growth retardation, aggression, weight loss, and other physical and mental health problems, especially in children. Numerous studies have been performed on intestinal parasitic infections in Iranian preschool and school children. However, no study has gathered and analyzed this information systematically. The aim of this study was to provide summary estimates for the available data on intestinal parasitic infections in Iranian children. We searched 9 English and Persian databases, unpublished data, abstracts of scientific congresses during 1996-2015 using the terms intestinal parasite, Giardia, Cryptosporidium, Enterobiusvermicularis, oxyure, school, children, preschool, and Iran. We conducted meta-analysis using STATA, and for all statistical tests, p-value less than 0.05was considered significant. Among the 68,532 publications searched as a result, 103 were eligible for inclusion in the study. The prevalence rate of intestinal parasitic infections was 38% (95% CI- 33%, 43%). Prevalence of protozoa, helminthic intestinal infections, and non-pathogenic parasites was 16.9%, 9.48%, and 18.5%, respectively, which affected 14.27% males and 15.3% females. The rate of infection in preschool and school children was 38.19% and 43.37% respectively. Giardia, Enterobiusvermicularis and Entamoeba coli were the most common among protozoa, helminthic, and non-pathogenic infections (15.1%, 16.5%, and 7.1%, respectively). The data analyses indicated that the prevalence of intestinal parasitic infection is decreasing in Iranian preschool and school children. Improvement of sanitation, personal hygiene, increased awareness of people, seasonal variations, and health education can be effective in reducing parasitic infections in different communities.