Detection and molecular identification of Entamoebaspecies in faecal samples from Duhok province, Kurdistan Region, Iraq.

The study involved the estimation of the prevalence of Entamoeba spp. using microscopy and molecular techniques among symptomatic outpatients during April 2021 to March, 2022. Stool samples were collected from 2592 outpatients with amoebiasis symptoms of both sexes and different ages (≤ l to 60). Also, 107 stool samples were taken randomly from asymptomatic individuals and examined microscopically to detect infection with Entamoeba spp. the positive specimens were used for molecular analysis with positive symptomatic samples targeting the 18S rRNA gene by nested PCR. Microscopically 21.68% (562/2592) were positive, for Entamoeba spp. Males showed highest infection rate than females (67.43% vs 32.56%). Ages from 1-10 years showed the highest rate (54.09%), and urban inhabitant had somewhat a higher rate than rural one (58.54% vs 41.45%) which was statistically non-significant(P>0.05). Among asymptomatic individuals, 57% (61/107) were positive for Entamoeba spp. Nested PCR analysis yielded 73% positive samples for Entamoeba spp. with a fragment size of 897 bp. Three fragment sizes were produced, for E. histolytica, E. dispar and E. moshkovskii which were 439, 174 and 553 bps, respectively. Single infection occurred with, E. histolytica in 46%, of symptomatic and 6% of asymptomatic cases, E. dispar in 38% of asymptomatic and 10% of symptomatic cases, E. moshkovskii, reported at very low rate among both groups.

Prevalence of amoebiasis and associated risk factors among population in Duhok city, Kurdistan Region, Iraq.

INTRODUCTION: Entamoeba histolytica, a protozoan parasite, is the third major contributor to human mortality and morbidity outside of malaria and schistosomiasis. The purpose of this cross-sectional study was to estimate the prevalence of Entamoeba spp. among outpatients of two teaching hospitals in Duhok city who agreed to participate in the study from April 2021 to March 2022 to assess the impact of associated risk variables on the infection rate. METHODOLOGY: Stool specimens were collected from outpatients suffering from diarrhea and other gastrointestinal symptoms in two teaching hospitals: Azadi and Heevi Pediatric in Duhok city, Kurdistan Region- Iraq. The collected stool specimens were examined macroscopically, followed by microscopic examination using the direct wet mount and zinc sulfate flotation methods, respectively. RESULT: Infection with Entamoeba species was recorded in 21.68% (562/2592) of the analyzed specimens. Males had a significantly higher infection rate than females (67.43% vs. 32.56%). This difference was statistically significant (p < 0.000). The highest rate was seen in the age group 1-10 years (p < 0.001). Lower levels of education, low incomes, eating unwashed fruits and vegetables, drinking well water, eating frequently outside of homes, not using antidiarrheal medications and living in overcrowded families were risk factors that showed high levels of infection (p < 0.0001). CONCLUSIONS: The present study concluded that improving living conditions, providing clean water, and promoting health education programs are essential to reduce the rate of this disease among the population.

Insights into Human Cystic Echinococcosis in the Kurdistan Region, Iraq: Characteristics and Molecular Identification of Cysts.

Cystic echinococcosis (CE) is a neglected worldwide distributed parasitic disease caused by the Echinococcusgranulosus sensu lato (s.l.) species complex. For a better understanding of the pathways of transmission of this parasite, clinical and molecular epidemiological studies are particularly needed from endemic areas where data are scant, such as in the Middle East. The study aimed to identify the characteristics, location, cyst stage and species/genotypes of E. granulosus s.l. complex in humans from the Kurdistan region, Iraq. To this aim, from June 2019 to February 2021, 64 echinococcal cysts were surgically removed from 62 patients in Azadi and Vajeen reference Hospitals at Duhok city, Duhok governorate (Kurdistan region, Iraq). The results confirmed the liver as the most common anatomical site of CE with 72.58% of the cases, followed by the lungs in 19.35%, while 66.13% of CE cases were females. The highest rate of infections occurred in the age class 21−30 (27.42%). High rates of CE were reported among patients living in rural areas and housewives, which were 54.84% and 43.55% of the CE patients, respectively. The fertility of echinococcal cysts was 82.81%, and the viability of fertile protoscoleces was 70.53%. Cysts were staged with ultrasound according to the WHO-IWGE classification as 32.8% CE1, 32.8% CE2, 7.8% CE3a, 9.4% CE3b, 15.6% CE4 and 1.6% CE5. Molecular analyses using mitochondrial NAD5 gene showed that all analyzed samples (n = 59) belonged to the genotypes G1 or G3 of E. granulosussensu stricto (s.s.), thus, confirming sheep−dog−human transmission in the Kurdistan region, Iraq. No statistically significant correlation was found between the genotypes G1−G3 of E. granulosus s.s. and variables, such as the fertility, location and cyst stage classification. Based on the present findings, it is necessary to implement monitoring and control programs in sheep and dog populations to decrease the odds of human infections. Public health education campaigns are required to be implemented at the community level to reduce the risk of acquiring CE in humans in the Kurdistan region, Iraq.

Two haplotype clusters of Echinococcus granulosus sensu stricto in northern Iraq (Kurdistan region) support the hypothesis of a parasite cradle in the Middle East.

Human cystic echinococcosis (CE) caused by Echinococcus granulosus s.s. is a major public health problem in Iraqi Kurdistan with a reported surgical incidence of 6.3 per 100,000 Arbil inhabitants. A total of 125 Echinococcus isolates retrieved from sheep, goats and cattle were used in this study. Our aim was to determine species/genotypes infecting livestock in Iraqi Kurdistan and examine intraspecific variation and population structure of Echinococcus granulosus s.s. in this region and relate it to that of other regions worldwide. Using nucleotide sequences of the mitochondrial cytochrome c oxidase subunit 1 (cox 1) we identified E. granulosus s.s. as the cause of hydatidosis in all examined animals. The haplotype network displayed a double-clustered topology with two main E. granulosus s.s. haplotypes, (KU05) and (KU33). The 'founder' haplotype (KU05) confirmed the presence of a common lineage of non-genetically differentiated populations as inferred by the low non-significant fixation index values. Overall diversity and neutrality indices indicated demographic expansion. We used E. granulosus s.s. nucleotide sequences from GenBank to draw haplotype networks for the Middle East (Iran, Jordan and Turkey), Europe (Albania, Greece, Italy, Romania and Spain), China, Mongolia, Russia, South America (Argentina, Brazil, Chile and Mexico) and Tunisia. Networks with two haplotype clusters like that reported here for Iraqi Kurdistan were seen for the Middle East, Europe, Mongolia, Russia and Tunisia using both 827bp and 1609bp cox1 nucleotide sequences, whereas a star-like network was observed for China and South America. We hypothesize that the double clustering seen at what is generally assumed to be the cradle of domestication may have emerged independently and dispersed from the Middle East to other regions and that haplotype (KU33) may be the main haplotype within a second cluster in the Middle East from where it has spread into Europe, Mongolia, Russia and North Africa. Further studies using metacestodes of human origin are required to investigate the biological importance of E. granulosus s.s. haplotypes/clusters and their association, if any with clinical manifestations of CE infection.