Genotype Distribution of Enterobius vermicularis Isolates from Northwest Provinces of Iran.

Background: Human infection with Enterobius vermicularis occurs worldwide. The most common clinical manifestation of a pinworm infection is an itchy anal region. This parasite is incidentally found in appendicitis. This study aims to characterize and genotype this parasite from different samples inferred by mt-DNA. Methods: Forty appendectomies for acute clinical appendicitis, 40 positive scotch-tape samples, and 10 adult females worm isolated from patients. Genetic differentiation, haplotype differences, and isolates population structure were analyzed based on the cytochrome c oxidase subunit I (cox1) gene. Results: It has been demonstrated that all isolations in the appendectomies specimens are similar, and the genetic difference divergence is seen in adult worm specimens. The neutral indices of the samples did not show a significant difference and show that there is no intra-specific and population distribution diversity. Conclusion: Our results show different haplotypes in the B type of E. vermicularis population and add new information about genotyping of these parasites in Iran. In comparison with other studies, intra-specific variation of this parasite from Iran was observed.

Morphological and molecular characterization of Fasciola isolates from livestock in Golestan province, northern Iran.

BACKGROUND: Fascioliasis, caused by the liver flukes Fasciola hepatica and Fasciola gigantica, is a global zoonotic helminthic disease. The livestock and human are the final hosts of the parasites. Northern Iran is an important endemic region for fascioliasis. Few studies have been conducted on the characterization of Fasciola isolates from eastern regions of the Caspian littoral of the country. OBJECTIVE: The aim of the present study was to identify F. hepatica, F. gigantica and intermediate/hybrid forms of Fasciola isolates from livestock in Golestan province, northern Iran, using morphometric and molecular tools. METHODS: Livestock livers naturally infected with Fasciola spp. were collected from Golestan slaughterhouse during 2019-2020. The worms were morphometrically studied using a calibrated stereomicroscope. Genomic DNA was extracted from all samples, and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed on internal transcribed spacer (ITS1) region using Rsa1 restriction enzyme. All the isolates were then analysed by multiplex PCR on Pepck region. RESULTS: A total of 110 Fasciola isolates were collected from the infected livers, including 94 sheep, 12 cattle and 4 goats. Morphometric analysis of 61 adult Fasciola isolates indicated that, 44 and 17 isolates belonged to F. hepatica and F. gigantica, respectively. Eighty-one and 29 isolates belonged to F. hepatica and F. gigantica using ITS1-RFLP, respectively. However, Pepck Multiplex PCR indicated 72 F. hepatica, 26 F. gigantica and 12 intermediate/hybrid forms. All 12 hybrid isolates were found in sheep host. Two isolates were identified as F. gigantica using morphometry and F. hepatica using both molecular methods. CONCLUSION: The present study confirmed the existence of both F. hepatica and F. gigantica species and reported the first molecular evidence of hybrid Fasciola isolates in ruminants of Golestan province.

Systematic Review and Meta-analysis of Role of Felids as Intermediate Hosts in the Life Cycle of Neospora caninum Based on Serological Data.

PURPOSE: Neosporosis is an important widespread parasitic infection caused by N. caninum. It infects a wide range of warm-blooded animals as intermediate hosts and dogs as the definitive host. Nevertheless, there are a number of questions regarding the life cycle and epidemiological aspects of N. caninum. Also, the role of felids (domestic and non-domestic) in the life cycle of N. caninum has been little described. Therefore, this study was conducted to evaluate the global prevalence of N. caninum in domestic and wild felids. METHODS: PubMed, Scopus, Google Scholar, Web of Science, and ScienceDirect databases were searched for articles published on the prevalence of N. caninum in felids until Aprill 2, 2022 and the reference lists of retrieved articles were screened. A random-effects meta-analysis model was used to estimate the pooled prevalence and 95% confidence interval. Heterogeneity among studies was evaluated using Cochran's Q and the I2 statistic. RESULTS: After exclusion of irrelevant articles and duplication removal, 30 studies were eligible for quantitative analysis including 20 studies on domestic cats and 10 studies on wild felids. The overall prevalence of neosporosis infection in cats was 15% (95% CI 10-21%) that was significantly higher in wild felids (26%, 95% CI 13-38%) than in domestic cats (11%, 95% CI 6-16%) (P = 0.03). There was no significant difference in pooled prevalence between male and female domestic cats (P = 0.75). Regarding continent, the lowest prevalence of neosporosis infection was in Asia (9%, 95% CI 1-20%) and the highest was in North America (43.6%, 95% CI 33.9-53.2%) and Africa (18%, 95% CI 9-46%). Higher prevalence was obtained when using the NAT with 22% (95% CI 7-37%), compared to the IFAT (17%, 95% CI 9-24%) and ELISA (6%, 95% CI 2-9%) (P = 0.01). CONCLUSION: The findings highlighted the importance of felids as potential intermediate hosts of neosporosis despite the fact that the source of the parasite for these animals was unknown. Further studies should be performed to investigate the role of this top predator (felids) in maintaining both domestic and sylvatic cycles of Neospora caninum.

Assessment of genetic markers for multilocus sequence typing (MLST) of Fasciola isolates from Iran.

BACKGROUND: Several markers have been described to characterise the population structure and genetic diversity of Fasciola species (Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica). However, sequence analysis of a single genomic locus cannot provide sufficient resolution for the genetic diversity of the Fasciola parasite whose genomes are ∼1.3 GB in size. OBJECTIVES: To gain a better understanding of the gene diversity of Fasciola isolates from western Iran and to identify the most informative markers as candidates for epidemiological studies, five housekeeping genes were evaluated using a multilocus sequence typing (MLST) approach. METHODS: MLST analysis was developed based on five genes (ND1, Pepck, Pold, Cyt b and HSP70) after genomic DNA extraction, amplification and sequencing. Nucleotide diversity and phylogeny analysis were conducted on both concatenated MLST loci and each individual locus. A median joining haplotype network was created to examine the haplotypes relationship among Fasciola isolates. RESULTS: Thirty-three Fasciola isolates (19 F. hepatica and 14 F. gigantica) were included in the study. A total of 2971 bp was analysed for each isolate and 31 sequence types (STs) were identified among the 33 isolates (19 for F. hepatica and 14 for F. gigantica isolates). The STs produced 44 and 42 polymorphic sites and 17 and 14 haplotypes for F. hepatica and F. gigantica, respectively. Haplotype diversity was 0.982 ± 0.026 and 1.000 ± 0.027 and nucleotide diversity was 0.00200 and 0.00353 ± 0.00088 for F. hepatica and F. gigantica, respectively. There was a high degree of genetic diversity with a Simpson's index of diversity of 0.98 and 1 for F. hepatica and F. gigantica, respectively. While HSP70 and Pold haplotypes from Fasciola species were separated by one to three mutational steps, the haplotype networks of ND1 and Cyt b were more complex and numerous mutational steps were found, likely due to recombination. CONCLUSIONS: Although HSP70 and Pold genes from F. gigantica were invariant over the entire region of sequence coverage, MLST was useful for investigating the phylogenetic relationship of Fasciola species. The present study also provided insight into markers more suitable for phylogenetic studies and the genetic structure of Fasciola parasites.

Molecular and phylogenetic analysis of E. vermicularis in appendectomy specimens from Iran.

Human infection with Enterobius vermicularis occurs worldwide, particularly in children. The role of E. vermicularis in appendicitis is neglected. This study was designed to investigate genotypes of E. vermicularis detected from appendectomy specimens in the human population from Iran and clarify the intra-species variation of the parasite. Seventy appendectomies for acute clinical appendicitis isolates from Azerbaijan and North Khorasan of Iran were used in the present study. The genetic information of Tehran and Hamedan regions was also obtained from GenBank for comparison and analysis. The nucleotide sequence of cytochrome c oxidase subunit 1 (cox1) gene was analyzed to perform genetic differentiation, haplotype network analysis, and population structure. Phylogenetic analysis of all the isolates were included in type B haplogroup. The number of haplotypes in all geographical locations of Iran is not much. Network analysis of sequences for regions such as Thailand, Iran, Denmark, and Poland show three classified subtypes B1, B2, and B3 in the B haplogroup. It seems that the haplotypes of E. vermicularis detected from appendectomy are B type, and divided into three subtypes. Further research using another genetic marker is required to elucidate the genetic variation of the parasites in detail.

Iranian Hydatid Disease Registry: Establishment and Implementation of a Neglected Tropical Disease Registry.

BACKGROUND: Cystic echinococcosis (CE) or hydatid disease is a global public health concern which imposes considerable economic costs on the communities in endemic regions. CE surveillance data are not adequately reliable. The present study reports the development and outcomes of a CE registry in Iran. METHODS: Hydatid Registry (HydatidReg) was initially established as a single-center registry in 2014 after the ethical approval of KMU. Following a call from MoHME to promote registry of different diseases and health outcomes, a call for participation was announced and all the Iranian Universities of Medical Sciences were requested to contribute to the registry. Subsequently, a nation-wide registry of hydatid disease was established in 2016. With a global perspective, HydatidReg joined the European Register of Cystic Echinococcosis (ERCE). A data collection form based on minimum dataset was designed and standard operating procedures (SOPs) were prepared to ensure standardized patient enrolment in the registry. A biobank system with two-dimensional barcoding was established along with HydatidReg for management and organization of biological specimens. RESULTS: As of March 2021, a total of 690 patients were enrolled in the registry. HydatidReg registered 362 (17.3%) out of the total 2097 patients enrolled in ERCE. Quality control (QC) of the data demonstrated 91.2% completeness and 80% timeliness. In the biobank, 322 biological specimens from 184 CE patients have been deposited including 70 blood, 96 sera and 156 parasite materials. CONCLUSION: High-quality data in the HydatidReg registry provided opportunities for health professionals to improve quality of care and organize meaningful research.

Three cases of brain hydatidosis in North Khorasan, Iran.

Cystic hydatidosis is a serious public health problem in Iran. Although cysts can develop in almost all organs and the brain cysts are very rare. Here, we present 3 confirmed cases of brain hydatidosis and the patients who underwent successful surgery. Pathological examinations demonstrated the presence of cystic hydatidosis.

SARS-CoV-2 detection in hospital indoor environments, NW Iran.

This study aimed to investigate the potential contamination of SARS-CoV-2 in indoor settled dust and surfaces of Amir Al-Muminin hospital in Maragheh, Iran. Samples were taken from surfaces and settled dust using a passive approach and particulate matter (PM) using an active approach from different hospital wards. SARS-CoV-2 was detected in 15% of settled dust samples (N = 4/26) and 10% of surface samples (3/30). SARS-CoV-2 has been detected in 13.8% and 9.1% of the dust samples collected at a distance of fewer than 1 m and more than 3 m from the patient bed, respectively. SARS-CoV-2 was found in 11% of surface samples from low-touch surfaces and 8% from high touch surfaces. The relationship between PM2.5, PM10, humidity, temperature, and positive samples of SARS-CoV-2 was investigated. A positive correlation was observed between relative humidity, PM2.5, and positive SARS-CoV-2 samples. Principal component analysis (PCA) suggested positive correlation between positive SARS-CoV-2 samples, relative humidity, and PM2.5. Risk assessment results indicated that the annual mean infection risk of SARS-CoV-2 for hospital staff with illness and death was 2.6 × 10-2 and 7.7 × 10-4 per person per year. Current findings will help reduce the permanence of viral particles in the COVID 19 tragedy and future similar pandemics e.g., novel influenza viruses.

Molecular and serological evaluation of visceral leishmaniasis in domestic dogs and cats in Maragheh County, north-west of Iran, 2018-2021.

OBJECTIVE: Zoonotic visceral leishmaniasis (VL) is caused by Leishmania infantum, of which dogs are the main reservoir. VL is endemic in the Middle East, also in some parts of Iran. Following reports of new cases of VL in children in Maragheh County, the non-endemic area of the disease, we encouraged to conduct a preliminary study on domestic dogs and cats to identify their potential role as reservoirs for the disease. MATERIALS AND METHODS: This study was conducted during a period of 3 years from 2018 to 2021. Two hundred ownership dogs and 25 cats from Maragheh County, north-west of Iran, were randomly screened. Blood samples were collected. A direct agglutination test (DAT) was used for the detection of anti-L. infantum antibodies. Furthermore, buffy coat samples from the L. infantum seropositive animals were examined to detect parasite presence using polymerase chain reaction. RESULTS: Out of the total of 200 ownership dogs evaluated, 170 (85%) were male and 30 (15%) were female with a mean age of 4.3 years. Anti-L. infantum antibodies (IgG cut-off ≥ 1:320) were observed in 3.5% of dogs (7/200) by the DAT test. All seropositive dogs were identified in the first year of examination. Regarding molecular approaches in seropositive dogs, two samples were positive for a 565 bp kDNA minicircle gene specific for L. infantum. During the study period, no seropositive case was detected in the cats examined. CONCLUSIONS: This study shows that the domestic cycle of L. infantum has been established in the studied region. It is necessary to increase the awareness and monitoring of the disease with the study of wild reservoirs periodically.

Molecular Identification of Fasciola Isolated from the Liver of Meat Animals in Fars Province, Iran.

Background: Fasciola hepatica and Fasciola gigantica are flatworms that infect animals and humans. Fasciola is the parasite of the liver or bile ducts and intestines of mammals, where such animals are known as their "definite hosts." The study aims to detect the genotype of Fasciola spp. from the livers of meat animals by using RFLP-PCR in samples collected from Fars province. Methods: Sixty Fasciola spp. samples were collected from infected slaughtered animals in three counties of Fars province, Iran (Jahrom, Nourabad Mamasani, and Kazeroun).Genomic DNA was extracted by the conventional phenol-chloroform method. For the study, PCR-RFLP and sequence analysis of the first nuclear ribosomal internal transcribed spacer (ITS1) region from Fasciola species were used to conduct the study. Results: The fragment of about 700 bp in all the Fasciola samples was amplified. In total, 43 samples of Fasciola gigantica and 17 samples of Fasciola hepatica were identified. Conclusion: The dominant Fasciola species in this region is Fasciola gigantica. Hence, it seems that hygienic policies should be developed to prevent and control fascioliasis because of the dominant species, Fasciola gigantica.

Comprehensive investigation of SARS-CoV-2 fate in wastewater and finding the virus transfer and destruction route through conventional activated sludge and sequencing batch reactor.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA transmission route was thoroughly investigated in the hospital wastewater, sewage collection network, and wastewater treatment plants. Samples were taken on four occasions from December 2020 to April 2021. The performance of two different wastewater treatment processes of sequencing batch reactor (SBR) and conventional activated sludge (CAS) was studied for virus destruction. For this purpose, liquid phase, solid phase and bioaerosol samples were taken from different units of the investigated wastewater treatment plants (WWTPs). The results revealed that all untreated hospital wastewater samples were positive for SARS-CoV-2 RNA. The virus detection frequency increased when the number of hospitalized cases increased. Detection of viral RNA in the wastewater collection system exhibited higher load of virus in the generated wastewater in areas with poor socioeconomic conditions. Virus detection in the emitted bioaerosols in WWTPs showed that bioaerosols released from CAS with surface aeration contains SARS-CoV-2 RNA posing a potential threat to the working staff of the WWTPs. However, no viral RNA was detected in the bioaerosols of the SBR with diffused aeration system. Investigation of SARS-CoV-2 RNA in WWTPs showed high affinity of the virus to be accumulated in biosolids rather than transporting via liquid phase. Following the fate of virus in sludge revealed that it is completely destructed in anaerobic sludge treatment process. Therefore, based on the results of the present study, it can be concluded that receiving water resources could not be contaminated with virus, if the wastewater treatment processes work properly.

Bionomics and phylo-molecular analysis of Leishmania species isolated from human lesions using ITS1 genes in north-east of Iran.

Leishmaniasis is a zoonotic infectious disease caused by Leishmania species. The identification of parasite species and the type of disease is beneficial for treatment and preventive modalities. Leishmania tropica and L. major have been reported as the main etiological agents of cutaneous leishmaniasis (CL) in Iran. The incidence of zoonotic CL has increased and different in distinct loci of Iran. Hence, we perused the Leishmania species and its genetic traits in the North East of Iran. The investigation was conducted on 200 positive smears prepared from patients' lesions suffering from CL referred to the health care centers of northeastern provinces in Iran from 2013 to 2019. The obtained positive microscopy samples were divided to score the ranges from + 1 to + 6, of them 40 smears exhibited low-parasitemia. Leishmania species analyzed using PCR-RFLP, genetic diversity indices evaluation, phylogenetic analysis, and sequencing comparison with other species in the GeneBank based on ITS1 gene. The isolated L. major strains were similar to other Iranian isolates in this region. Pairwise fixation index (FST) index was statistically significant in different L. major populations and showed the genetic differences in pairwise population of different geographical locations of Iran. The current study confirmed an old pattern endemicity of zoonotic CL in North-east of Iran. Therefore, in order to assess the hybrid formation, more epidemiological, ecological, and gene polymorphism studies are needed to understand the pathogenic role of Leishmania species in Iran.

Prevalence of human visceral leishmaniasis in Iran: A systematic review and meta-analysis.

Leishmania infantum is the main cause of human visceral leishmaniasis (HVL; also known as kala-azar) in the Middle East and may be fatal if left untreated. This disease was first reported in 1949 in Iran. Despite marked improvements in hygiene and sanitation conditions, the disease is still endemic in some parts of Iran. It is difficult to determine the current prevalence of HVL in Iran due to the scarcity of comprehensive studies in this regard. In response to this gap, a systematic review and meta-analysis was conducted to gain better understanding of HVL epidemiology in the general population of Iran. English and Persian databases were searched for studies reporting the prevalence and risk factors associated with HVL in the Iranian people from January 1995 to December 2019. The reported data were selected according to inclusion and exclusion criteria. The pooled prevalence of HVL infection and its 95 % confidence intervals were calculated. Quality assessment, heterogeneity testing and publication bias assessment were also done. Literature search revealed 3634 studies, of which 35 studies met our eligibility criteria, with a total of 50,716 individuals. The meta-analysis was performed on 31 out of 35 included studies. The estimated pooled prevalence of HVL infection according to seropositivity was 2% (95 % CI: 1-2%) in the general population of Iran in which western and northern provinces had the lowest and the highest prevalence, 0.5 % (95 %CI, 0.2-0.7%) and 3% (95 %CI, 1-5 %), respectively. The seroprevalence of HVL among females (2%; 95 %CI, 1-2 %) was more than males (1%; 95 %CI, 1-2 %). The ≤10 and >10 years age groups had similar seroprevalence rates (1%, 95 %CI, 1-2 % versus 1%, 95 %CI, 0-1 %, respectively). There was no significant difference in terms of geographic area, age and sex. Of 31 studies included in the meta-analysis, the most common diagnostic test was the direct agglutination test (96.77 %). To the best of our knowledge, this is the first systematic review of the prevalence of HVL in Iran. The results showed a low seroprevalence of HVL infection. However, the lack of published reports of HVL in an area does not exclusively mean the absence of the disease or carrier. We therefore recommend further studies in this regard.

Biodiversity Indices and Medically Importance of Ticks in North Khorasan Province, Northeast of Iran.

BACKGROUND: Ticks are considered as the main vectors for the transmission of various pathogens such as relapsing fever and CCHF to humans. This study was investigated the biodiversity indices and medically importance of ticks in North Khorasan Province, Northeast of Iran during 2015-2019. METHODS: Specimens were captured from infested ruminants including cows, sheep, and goats. Additionally, tick collections also were performed on non-domesticated creatures such as turtles, rodents, and hedgehogs. Specimens were identified using valid identification keys. Species diversity, species richness and evenness indices have been calculated to estimate species biodiversity of ticks. RESULTS: A total of 1478 adult ticks were collected. The specimens were from two families: Ixodidae (90.05%) and Argasidae (9.95%), 6 genera and 17 species including: Rhipicephalus sanguineus (55.9%), Rhipicephalus bursa (13.4%), Hyalomma marginatum (9.5%), Hyalomma anatolicum (9.5%), Hyalomma asiaticum (0.2%), Hyalomma aegyptium (0.5%), Hyalomma scupense (1.3%), Hyalomma sp (1.2%), Haemaphysalis sulcata (0.7%), Haemaphysalis erinacea (0.1%), Haemaphysalis inermis (0.1%), Haemaphysalis punctata (0.2%), Haemaphysalis concinna (0.1% Boophilus annulatus (1.2), and Dermacentor marginatus (6.1%) among hard ticks as well as Argas persicus (91.8%) and Argas reflexus (8.2%) amongst soft ticks. Rhipicephalus sanguineus, Rh. Bursa, Hy. marginatum and Hy. anatolicum were known as the most frequent species of hard ticks. Tick's species richness, Shannon diversity index and Simpson index in this area were S= 17, H'= 1.69, D= 0.294 respectively. CONCLUSION: Based on tick distribution veterinary authority, public health organizations and other officials should act for implementation of disease prevention.

Molecular phylogenetic and genetic variability of Fasciola gigantica in Kermanshah province, western Iran with an overview to understand haplotypes distribution in Asia and Africa.

Over the last decade, diagnostic tools to detect and differentiate Fasciola species have improved, but our understanding of the distribution of haplotypes and population structure of this parasite is less clear. This study was designed to survey this gap in the F. gigantica epidemiology in Kermanshah province, western Iran from 2015 to 2017. Sixty-eight Fasciola isolates were collected from slaughterhouses from this province. We evaluated the PCR-RFLP assay of the ITS1 genes for the identification of Fasciola species using the RsaI enzyme. After Fasciola species identification, the partial sequence of mitochondrial NADH dehydrogenase subunit 1 (ND1) gene of F. gigantica was used for subsequent construction of the phylogenetic tree and network analysis. Based on the PCR-PRFLP profile, one (6.25%) of sheep isolates and 19 (39.60%) of cattle isolates were detected as F. gigantica, whereas 93.75% of sheep isolates, 60.40% of cattle isolates and all of the goat isolates were F. hepatica. In the 20 analyzed flukes, five ND1 haplotypes were detected. Statistically significant genetic differentiation was demonstrated between the Iran population and all the other populations. Evidence is presented for the existence of two well-separated populations: African and West Asian gigantica flukes and East Asian gigantica flukes. Genetic relationships among haplotypes were associated with geographical divisions. Also, our results have heightened our knowledge about the genetic diversity of F. gigantic, providing the first evidence for the existence of two well-separated populations of this parasite.

Genetic diversity analysis of Blastocystis subtypes and their distribution among the domestic animals and pigeons in northwest of Iran.

Blastocystis is a unicellular, anaerobic, eukaryotic protist, a common parasite found in the intestinal tracts of animals and humans. During the last few years, the host fecal DNA analysis by nucleic acid-based method has led to significant advances in Blastocystis diagnostics and enabled subtypes (STs). The zoonotic transmission of Blastocystis to humans is not well understood, therefore the present study was conducted to identify Blastocystis subtypes in Iran from different animal hosts from northwest of Iran. A total of 427 fresh fecal specimens were collected from cattle, sheep, poultry and pigeon (40,150,132,105 respectively). To detect the Blastocystis sp., each fecal specimen was examined microscopically. Total DNA from the samples that were positive for Blastocystis sp. was isolated, and the barcoding region of the small subunit of ribosomal rRNA (18S rRNA) was amplified and sequenced. Subsequently, sequence analyses, genetic diversity indices and evolutionary relationships of Blastocystis subtype populations were carried out. In total, 14.98% of the analyzed samples were positive for Blastocystis sp. and the subtypes detected were ST3,7,10 and 14. Among these, the ST10 was the main subtype that was found only in the cattle, sheep and poultry and the zoonotic subtype ST3 was present only from cattle. Our study shows the presence of Blastocystis subtypes in the sheep in north west of Iran and also demonstrated that the genetic approaches are crucial to understand the host specify of subtypes and the mode of infection. The study suggests that the genetic approaches will help us to understand the host specificity of subtypes and their role in infection if they are obtained from human and animals from the same geographical locations. Therefore, it is important to study the zoonotic aspects of this parasite with large number of samples from different groups of animals and from different geographical locations.

Clinico-Hematological Findings of Acute Pediatric Visceral Leishmaniasis Referred to the Northeast of Iran during 2005-2015.

BACKGROUND: To characterize the epidemiological, clinical, hematological and biochemical features of 33 cases hospitalized with pediatric visceral leishmaniasis (PVL) in North Khorasan Province of Iran from 2005 to 2015. METHODS: The serological, hematological and biochemical tests were employed in 33 children between 8 months to 6 yr with a final diagnosis of acute visceral leishmaniasis (VL). The diagnosis of VL was established by microscopic demonstration of Leishmania spp. amastigotes inactive bone marrow aspiration (BMA). RESULTS: The most common presenting features were anemia (82.5%), fever (75%), and hepatosplenomegaly (45.4%). Various hematological parameters showed that most patients were suffering from moderate to severe microcytic hypochromic anemia (78.8% had RBC count less than 4 million cells/ul, 67.7% Hb less than 8 fl). 66.7% of them were leukopenic (WBC: less than 5× 10 3 /µL) and 24.2% had decreased platelet counts. Pancytopenia was observed in 18.2% of cases. MCV, MCH, and MCHC levels were below the reference range in 88%, 90% and 85.1% of the patients respectively. Moreover, aspartate transaminase (AST) and alanine transaminase (ALT) levels were increased in 53.33% and 6.66% of the patients respectively. 92.9% of cases were C-reactive protein (CRP) positive. Bone marrow was found hyper-cellular in all of them, and myeloid to erythroid ratio (M/E) was more than 4 in 39.1% of cases. Plasma cells slightly were increased in 60% of patients and megakaryocytes were decreased in thrombocytopenic patients. CONCLUSION: Bone marrow/splenic aspiration still remains the gold standard test despite its risk and pain for patients.

Cryopreservation of Echinococcus granulosus Protoscoleces.

BACKGROUND: The main objective of the current study was to investigate on the cryopreservation of protoscoleces of Echinococcus granulosus, a causative agent of cystic hydatidosis in man. METHODS: This study was conducted on isolated protoscoleces from hydatid cysts infected livers collected from slaughterhouse of Tehran, Iran in 2016. Viability of protoscoleces was evaluated by dye test. Cryopreservation of isolated protoscoleces in the presence of Dimethylsulfoxide (DMSO) and glycerol using a three-step cooling protocol involving an initial period at -20 °C, -80 °C and liquid nitrogen was performed. RESULTS: The mean viability rate of 10% DMSO and 15% glycerol were 9% and 8% respectively. The protoscoleces of Echinococcus granulosus have been successfully thawed and recovered after 6 months storage in liquid nitrogen. CONCLUSION: Cryopreservation method needs to be improved for each species of helminthes and can be useful for other immunological and laboratorial studies.

Genetic diversity and distribution of Fasciola hepatica haplotypes in Iran: Molecular and phylogenetic studies.

Fasciolosis is a zoonotic disease caused by Fasciola hepatica and Fasciola gigantica. Over the last decade, diagnostic tools to detect and differentiate Fasciola species have improved, but our knowledge of the distribution of haplotypes and gene flow of this parasite is not comprehensive yet. The purpose of this study was to investigate this gap in the epidemiology of F. hepatica in different provinces of Iran between 2015 and 2017. Isolated Fasciola were collected from abattoirs in 9 provinces. The partial sequence of mitochondrial NADH dehydrogenase subunit 1 (ND1) gene was used for the identification and molecular analysis of F. hepatica isolates. The amplified PCR products were purified and subjected to direct sequencing for subsequent construction of phylogenetic tree and network analysis. In the 130 subjects analyzed, 37 ND1 haplotypes were detected. This is the first study in Iran which investigates F. hepatica population and its genetic structure, based on mitochondrial ND1 marker in different geographical regions of Iran.