Giardia lamblia is a significant intestinal protozoan in humans worldwide, with high incidence of infection in developing countries, particularly among children. Eight assemblages (A to H) have been identified by molecular analysis; A and B are more frequently associated with human infections. Regardless its importance, to the best of our knowledge, this is the first molecular study on assemblages in Giardia lamblia adopted in Zakho district, province of Duhok, Iraq. The present study aimed to determine the giardiasis infection rate and identify the assemblages of Giardia lamblia in children from four areas in Zakho district. Fecal samples were collected, and microscopic examination was done. Genomic DNA was extracted, and assemblage identification was done via amplification of the gdh gene using a semi-nested polymerase chain reaction and restriction fragment length polymorphism. Out of 31 Giardia-positive samples, 23 were successfully amplified through semi-nested PCR. Nineteen isolates (82.60%) were assigned to assemblage B, and four(17.40%) to assemblage A. Assemblage B was identified as belonging to sub-assemblages B11 and B1V, while Assemblage A was identified as sub-assemblages A1 and A11. This study provides insights about Giardia lamblia assemblages in Zakho district, Duhok province, Iraq, and may serve as a beginning step toward understanding the molecular characterization of Giardia in the studied area.

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.

Sarcocystis is an intracellular parasite belonging to the phylum Apicomplexa. More than two hundred species of Sarcocystis are capable of infecting different species of wild and domesticated animals and humans. This study aimed to determine the prevalence of Sarcocystis species and their morphological characteristics in the esophagi muscles of ruminants slaughtered at Zakho abattoir, Zakho City, Kurdistan Region/Iraq. Esophagi muscle specimens’ of 1586 ruminants including, 764 sheep, 569 goats and 253 cattle were examined macroscopically and microscopically from April 2021to March 2022 to determine the prevalence of macroscopic and microscopic sarcocysts. Results revealed a total rate of infection with macrosarcocysts at 18.03 % (286 / 1586), with the highest rate 26.85% (254/946) in local breeds versus 5% (32/640) of imported ones. The highest rate among local breeds 90.91% (10/11) was in cattle, while the highest rate in imported breeds 9.13% (21/230) was among sheep. The overall rate in males was higher than in females. The detected macrosarcocysts relying on morphological characters were assigned as Sarcocystis gigantea, S. moulei, S. medusiformis, S. fusiformis, and S. hirsuta. Histological examination revealed the presence of microsarcocysts in the esophageal muscles of the three hosts which were assigned as S. tenella, S. arieticanis in sheep, S. capracanis in goats and S. bovifelis or S. hominis in cattle. The study results show the widespread of sarcocystosis among ruminants in this region that poses a threat to livestock and humans. This condition necessitates the development of preventive strategies to minimize or eradicate this parasitic infection.

Toxoplasmosis, an infectious disease caused by the obligate intracellular protozoan parasite, poses varying degrees of risk, ranging from asymptomatic cases in immunocompetent individuals to severe, life-threatening conditions in immunocompromised individuals and developing fetuses, especially when infection occurs during early pregnancy. While the disease is endemic in Iraq, there is a notable lack of precise information regarding its seroprevalence among females of childbearing age and pregnant women, along with associated risk factors in the Zakho district.

The Prevalence and Molecular identification of Cryptosporidium species among Human Population in Zakho District, Duhok Province, Kurdistan Region, Iraq. Ahmed Basheer Mohammed a*, Hamdea Ismail Ahmed a , Wijdan Mohammed Salih Meroa,b , a Department of Biology, Faculty of Science, University of Zakho, Duhok Province, Kurdistan Region, Iraq b College of Science, Nawroz University, Duhok Province, Kurdistan Region, Iraq * Corresponding author: Dr. Ahmed Basheer Mohammed E-mail: ahmed.mohammed@uoz.edu.krd Abstract This cross-sectional study was carried out to determine the prevalence and molecular identifications of Cryptosporidium species among the human population in Zakho district, Duhok Province, Kurdistan Region, Iraq. The study was performed during the period from August 2021 to March 2022, involved 700 fecal specimens of both genders and different ages collected from Zakho General Hospital and primary schools located in suburban area of Zakho city. The data were analyzed using statistical software version SPSS (25). Cryptosporidium species oocyst was found in 87.57% (613/700) of the examined specimens using Modified Ziehl Neelsen technique. Males showed slightly higher infection rate than females (88.4% vs 85.5%) with non-significant difference (P> 0.05). The highest infection rate (91.7%) was present in individual between the ages of >30 to 40 years, with statistically significant difference (P<0.05). Molecular identification of Cryptosporidium species was estimated using 18S-rRNA gene for Cryptosporidium genus using Nested PCR technique and genus specific primers (Cry18 AS1 and Cry18 S2) that yielded 347 bps product indicating Cryptosporidium genus, then species-specific (Cry18 AS1 and Cry18 S1) were used for the detection of Cryptosporidium parvum and Cryptosporidium hominis. 45/50 samples were positive and revealed 240 bps. The acquired sequences were deposited into the GeneBank database under the accession numbers OP107010 for C. hominis, OP104444, OP153870, OP163188, OP163414, and OP164631 for C. parvum.

In Iraq generally and Kurdistan region specifically, goats are an important resource for meat and milk production. It is well known that Myostatin genes (MSTN) have the essential role in growth and development due to its crucial involvement in muscle growth. The goal of this research was to reveal the polymorphism of MSTN gene in different Capra hircus breeds (native, Shami, Meriz and Kamori goat) and in wild mountain goat (Capra aegagrus) via PCR-RFLP technique and direct seqencing The MSTN (exon 1) polymorphisms were investigated in all breeds, MSTN-Dra I/RFLP revealed three genotypes AA and BB were homozygous and AB was heterozygous. The calculated genotype frequencies were 0.270, 0.368 and 0.362 for AA, BB and AB genotype respectively. The allelic frequency was 0.451 for the A allele and 0.549 for the B allele. The average of the observed heterozygosity was 0.362 and the observed homozygosity was 0.638. The sequence data of MSTN gene of Native and Shami goats revealed 31 SNPs in nonfunctional region.

Entamoeba histolytica is 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 out patients of two teaching hospitals in Duhok, Kurdistan Region, Iraq.

Entamoeba histolytica is 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 out patients of two teaching hospitals in Duhok, Kurdistan Region, Iraq.

Abstract: In Iraq, Awassi is the most widespread sheep breed of non-European origin. The breed is adapted to a wide range of environmental conditions. Sheep farming, mainly sheep of Awassi breed, represents an essential economic activity of Iraq; however, so far, only very few genetic data exist on this breed. The present study was performed in Biology Department, University of Zakho. A total of 32 blood samples were obtained from Awassi-Iq (Ovis aries) sheep breed in three farms located in Nineveh province, Iraq. Genomic DNA extraction was performed using spin column method. Specific marker (GenBank: NC001942) used for amplifying 550 bps fragments of mitochondrial DNA in PCR assay. The PCR products were separated by electrophoresis on 1.5% agarose gel, visualized by staining with RedSafe dye, and photographed by a digital camera. Ten PCR products were sequenced and compared to sequences from other countries available in NCB1.The main results indicate that Awassi- 1q (Ovis aries) sheep breed shares similarity to both Chines Tibetan sheep breed (Linzhou); Iranian sheep breed (91.37%), to both Iraqi sheep breeds Hamdani; Nima (91.62%) respectively and to Ovis aries AKK- 46 from Turkey (92.35%). However, Awassi-Iq was shown to be monophyletic, out group breed and diverged earlier than related breeds (Turkey, Iran, and China). This study highlights the importance of Mitochondrial DNA analysis in genetic study of sheep breeds.

Cryptosporidiosis caused by species belonging to genus Cryptosporidium. Cryptosporidiosis is a zoonotic disease that affects both animals and humans, making it a major public health and veterinary concern. A cross sectional study was performed during the period from August 2021 to March2022, to estimate the prevalence of Cryptosporidium spp. in the human population in order to determine the effect of associated risk factors on the rate of infection in Zakho district, Duhok. Seven hundred Fecal specimens were collected from Zakho General Hospital and two primary schools located in suburban area of Zakho city.

In Iraq generally and Kurdistan region especially, goats are important resource of meat and milk production. Growth hormone (GH) plays an important role in several biological processes such as reproduction, lactation, metabolism and growth of livestock animals. The objective of this study was to reveal the polymorphism of growth hormone gene (GH) in five goat breeds (Native, Shami, Merzi , Afghani and Wild goat) via PCR-RFLP technique. Two loci on GH gene named GH1 and GH2 polymorphisms were analyzed. . GH1- HaeIII / RFLP showed only two genotypes (AA and AB) in all breeds, with the absence of genotype BB. The genotype frequency was 0.882 for AB and 0.118 for AA genotype respectively; the allelic frequency was 0.492 for A allele and 0.508 for allele B. However, digestion of GH2 by HaeIII was monomorphic. PCR-RFLP could be employed as useful markers, helping goat breeder in selection of goats for high growth performance.

Fascioliasis caused by Fasciola species (Fasciola hepatica and Fasciola gigantica) is considered as the most important helminthic infection of domestic animals in developing countries. The present study was performed during January to December 2019, in Biology Department, University of Zakho. Hundred adult’s flukes of Fasciola spp. were isolated from bile ducts and gallbladder of cattle, sheep and goats slaughtered at different abattoirs (Duhok, Zakho, Shelidiza, Aqrah and Bardarash) in Duhok governorate, Kurdistan region, Iraq. Genomic DNA extraction was performed using a Genomic DNA Extraction kit, Jena Bioscience GmbH (Germany). Specific markers ITS1 and ITS2 ribosomal DNA were used for amplifying 480 and 550 bps fragments. DNA sequences have been used to characterize these liver flukes. The PCR products were separated by electrophoresis in 1.5% agarose gel, visualized by staining with RedSafe dye, and photographed. ITS-1 and ITS-1 markers were amplified successfully, the length of produced band for ITS-1 was 480 bp and for ITS-2 was 550 bp. Thirteen Fasciola spp. flukes isolated from cattle, sheep and goats from different districts were sequenced, from these thirteen PCR products, 7 amplicons were recognized as Fasciola hepatica and 6 amplicons as Fasciola gigantica. These sequences were deposited in GenBank under accession numbers: MW161261, MW084365, MW052601 MW084349, MW082585, MW084348, MW082830 for F. hepatica and MW161260, MW052602, MW084353, MW084350, MW082589, MW085034 for F. gigantica. This study confirms the prevalence of Fasciola hepatica and Fasciola gigantica in Iraq by using the ITS-1 and ITS-2 rDNA.

The study was performed on 504 stool samples from infants and children with diarrhea who attended laboratories at Zakho General Hospital, Heevie Pediatric Hospital, Chamisku and Bersive 1 camps, and children who did not have diarrhea in various primary schools in Zakho between August 2021 and the end of July 2022. The study focused on 1 month of age to 15 years old. The collected stool specimens were examined macroscopically followed by microscopic examination using wet mount and concentration methods. The microscopic findings revealed an overall prevalence of Giardia lamblia at a rate of 6.15% (31/504), with slightly but non-significantly higher infection rate in males than in females (6.71% vs. 5.43%). The highest rate of infection was noted in the age group >3-6 years, while the lowest rate was in the age group >12-15 years, which were 11.76% and 1.45%, respectively. The infection rate was also twice as high among rural residents as compared to urban ones (10.34% and 3.32%), respectively. Infants and children who drink tap water had a high infection rate, whereas those drinking bottled water had a lower infection rate (8.05% and 1.92%), respectively. The infection rate was non-significantly higher in families with more than 6 members than in those with less than 6 members (7.06% and 5.22%) respectively. In terms of monthly infection distribution, the highest prevalence was reported in July 11.29%, and the lowest was recorded in February (2.27%). Finally, the statistical analysis revealed a significant relationship between infection and age group, residency, as well as the types of drinking water consumed, but non-significant relationship between infection and gender, number of family members and months of sample collection.

One of the most common mycobiotic in human which colonizing several functional site like skin, mouth, urinary tract, vagina is Candida without causing any infection (Seneviratne, Jin and Samaranayake, 2008) Modification in local environment of the host enhanced candida growth and hence led to candidiasis (Pope and Cole, 2002). Virulence factors of candida as well as immune system of the host switching Candida form harmless commensal to disease causing pathogen (Yang, 2003). Most common virulence factors of candida; Adherence, extracellular hydrolase enzymes while help the candida to adapt to specific anatomical sites. The pathogenesis of Candida species enhanced by phenotypic switching, filamentation and biofilm formation.(Calderone, Fonzi and Fonzi, 2001). Extracellular enzymes such as phospholipases, protease, lipase and haemolysin contributed in the pathogenicity of Candida species. (Silva et al., 2009) Furthermore, the capability of C. albicans to get iron by haemolysin production is essential in its persistence and capability to form infections in the human’s body. Hemolytic activity is one of the virulence factor that facilitate hyphal invasion and disseminate candidiasis. Odds FC. Candida and Candidosis. 2. London, UK: Bailliere Tindall; 1988. The activity of Hemolysin by Candida is enhanced by present of glucose in the blood agar.(Manns, Mosser and Buckley, 1994). Hemolysins production by Candida albicans destroy hemoglobin in erythrocytes and obtain elemental iron. Hence, hemolysins considered as essential virulence factors which make pathogens to survive and persist (Luo et al., 2004).

Fascioliasis is considered as a neglected zoonotic disease of ruminants. This disease is caused by two species of Fasciola (F.) namely Fasciola hepatica and F. gigantica. They are distributed in more than 70 countries except Antarctica, in areas where sheep and cattle are reared (WHO 2016; CDC 2019). This disease was first recorded as early as 2000 BC. Fasciola spp. cause serious pathological effects to livestock (Abbas et al. 2019). The number of infected sheep exceeds 250 million and about 300 million cattle are infected globally with fascioliasis. The disease cause losses exceeding 3.2 billion $ yearly, due to decrease in milk and meat production and an increase in mortality rate in addition to liver condemnation and expenditure of anthelminthic (Hillyer and Apt 1997; Jaja et al. 2017).

Fascioliasis is one of the most common neglected zoonotic disease caused by Fasciola species. The current study was conducted during January to December 2019,in which five districts were selected in various parts of Duhok governorates which differ in their environmental and geographical conditions they included: Duhok city, Zakho, Shelidiza, Aqrah and Bardarash. Each slaughter house was visited once monthly to determine the rate of fascioliasis among domestic animals; cattle, sheep and goats. The infected livers were examined grossly and the bile duct and gall bladder were inspected for the presence of Fasciola species, the work was performed in Parasitology Laboratory, Biology Dept., Faculty of Science, Zakho University. Fascioliasis was diagnosed in 2.0% (195/9757) of total slaughtered livestock, with the highest rate of infection in cattle which was 3.27% (66/2021), followed by sheep 1.76% (122/6932) and the lowest in goats 0.87% (7/804). Slightly higher prevalence rate from positive cases was found in local livestock compared to imported one (2.10% versus 1.88%). According to gender, males showed higher prevalence than females (2.08% versus 1.08%). Regarding Fasciola species, among sheep and goats; the prevalence of F. hepatica was higher than Fasciola gigantica. While among cattle, F. gigantica was the most common species. With respect to parasite burden, the heaviest burden was recorded in 38.46% (20-100 flukes/liver) of slaughtered livestock with the heaviest being in sheep (55.74%), followed by goats (71.43%) while lightest burden was recorded in cattle in 62.12% (1-10 flukes/liver). Most of the liver flukes were recovered from the bile ducts (67.69 %), followed by gall bladder (21.54%) and the least (10.77%) from both the bile ducts and gall bladder.

A survey was carried out on the incidence of Candida yeasts in urine of healthy (non-diabetic) and diabetic women in Duhok and Zakho cities from February 2019 to April 2019. Out of 620 urine samples examined, 144 samples were found positive for Candida infection (23.2%). There were significant differences in incidence of Candida infection between urine of healthy and diabetic women as well as among age groups. All Candida isolates from positive samples were identified by using Chromogenic Candida Agar medium and confirmed by sequencing of the ITS1 and ITS4 region of rDNA. The identified Candida species were C.albicans, C.glabrata, C.krusei and C.tropicalis. The most common species was C.albicans (38.8%), followed by C.glabrata (34%), then C.tropicalis (13.8%), whereas, C.krusei displayed the least incidence (9.7%). All tested strains (n=83) for their hemolytic activity on Sabouraud's dextrose agar amended with sheep blood and glucose showed positive activity. Candida albicans and C.tropicalis displayed the highest hemolytic activity.

A total of 36 unrelated males from the two population groups in Iraqi Kurdistan: Kurds and Arabs were analyzed for eight Y-chromosome STRs (DYS19, DYS392, DYS437, DYS448, DYS456, DYS458, DYS635 and Y-GATA-H4). Total DNA from blood cells was extracted using DNA extraction Kit. A number of genetic parameters such as mean number of alleles, allele frequency, gene diversity, polymorphic information content (PIC), and genetic distance were calculated using Power Marker V3.25 software. The Dendrogram separated the populations into two main clades, the Kurd group and the Arab group except in one case only from the whole population.

Candida species are part of the body normal flora .Under certain conditions, these opportunistic microorganisms may lead to infection.The purpose of this study was to identify Candida species isolated from sputum from patients suffering from respiratory tract disorders.Methods:A total of 59 sputum samples taken from patients attending Azadi hospital at Duhok province ,Kurdistan Region/Iraq. For primary isolation,sputum samples were cultured on Sabouraud Dextrose agar (SDA).Suspected colonies of Candida isolates were then subcultured on Chromogenic Candida agar for presumptive identification. Genomic DNA extraction was performed using a Genomic DNA Extraction kit. For rapid identification of Candida spp, specific primers based on the genomic sequence of DNA topoisomerase 11 of C. albicans, C. parapsilosis I, C. parapsilosis II, C. guilliermondi,C. dubliniensis, C. krusei, C. kefyr and C. glabrata,C. tropicalis I, C. tropicalis II, C. lusitaniaewereused.TheMultiplex PCR products were separated by electrophoresis in 1.5% agarose gel, visualized by staining with ethidium bromide, and photographed .Results: ThreeCandida species namely C.albicans,C.glabrata and C.tropicalis were differentiated by their colour produced on Chromogenic Candida agar.PCR with the primer mixes yielded 4 different sized of PCR products corresponding to C.albicans, C.glabrata, C.Keyfer and C.tropicalisII.C.glabrata was the most common species (33.33%), followed by C.albicans (16.66%). The highest rate of isolation of Candida species was between the ages of 36 to 45.

Fascioliasis caused by the liver flukes of the genus Fasciola is considered as the most significant Trematodes infection of ruminants in both temperate and tropical countries. In the present study fifty adult Fasciola flukes (Platyhelminthes: Trematoda: Digenia) were collected from the bile ducts and gallbladder of infected bovine hosts (cattle) slaughtered at Duhok abattoir, Kurdistan region, Iraq. Genomic DNA extraction was performed using a Genomic DNA Extraction kit. ITS-1 and ITS-2 ribosomal DNA sequences have been used to characterize these liver flukes as a specific marker. The PCR products were separated by electrophoresis in 1.5% agarose gel, visualized by staining with ethidium bromide, and photographed. ITS-2 marker for F. hepatica was amplified successfully and the length of produced band for ITS-2 was 330 bp. The present study is the first trail for molecular characterization of F. hepatica in cattle in Duhok, Kurdistan Region, Iraq using the ITS-2 rDNA as a reliable genetic marker.

Candida species are the second most common cause of vulvovaginitis worldwide. The purpose of this study was to identify the species of vaginal Candida isolates by using phenotypic and Multiplex PCR techniques. 91 isolates from patients admitted to Azadi hospital and Maternity hospital in Duhok city were collected. The vaginal swab specimens were inoculated on Sabouraud Dextrose Agar. Colonies were then sub cultured on Chromogenic Candida agar. Genomic DNA extraction was performed using a Genomic DNA Extraction kit. For rapid identification of Candida spp, specific primers based on the genomic sequence of DNA topoisomerase 11 of C. albicans, C. parapsilosis I, C. parapsilosis II, C. guilliermondi,C. dubliniensis, C. krusei, C. kefyr and C. glabrata,C. tropicalis I, C. tropicalis II, C. lusitaniae were used. The multiplex PCR products were separated by electrophoresis in 1.5% agarose gel, visualized by staining with ethidium bromide, and photographed. 4 Candida species, namely C. albicans,C. glabrata,C. krusei and C. tropicalis were distinguished by Chromogenic Candida agar on the basis of colony colour and morphology .PCR with the primer mixes yielded 7 different sized of PCR products corresponding to C. albicans, C. guilliermondii, C. dubliniensis, C. glabrata, C. kefyr, C. krusei and C. tropicalis II. The analysis revealed C.glabrata and C. albicans were the most common species isolated with the percentage 40% and 30% respectively. This study concluded that phenotypic characteristics on selective agar medium such as Chromogenic Candida agar are useful for presumptive identification of Candiada spp with the support of molecular method such as Multiplex PCR.

In this study a molecular based technique was employed to characterize two Iraqi goat breeds . For this thirty blood samples were obtained from two indigenous goat breeds( Native and Merzi) in Duhok province. Genomic DNA was extracted , out of eight SSR primers , 6 Primers were amplified and produced bands.