Genetic diversity and haplotypes of Cysticercus tenuicollis isolates from slaughtered sheep and goats in Elazig and Bingol provinces of Turkey.

BACKGROUND: The cestode Taenia hydatigena uses canids, primarily dogs, as definitive hosts, whereas the metacestode larval stage cysticercus infects a range of intermediate hosts, including domestic animals such as goats, sheep and pigs. OBJECTIVES: In this study, we aimed to determine the genetic differences and haplotypes of Cysticercus tenuicollis isolates obtained from sheep and goats slaughtered in the Bingol and Elazig provinces of Turkey. METHODS: C. tenuicollis isolates were collected from 44 sheep and 26 goats slaughtered in slaughterhouses in Bingol and Elazig. After the isolation of total genomic DNA from C. tenuicollis isolates, the genetic characterization of the partial mitochondrial cytochrome oxidase 1 (CO1) gene region (866 bp) was amplified using specific primers by polymerase chain reaction (PCR), the products were then sequenced, and haplotype and genetic diversity analyses were carried out. RESULTS: As a result of the haplotype network analyses, 34 different haplotypes were detected around the main haplotype (Hap02) arranged in a star-like configuration and separated from other haplotypes by 1-28 mutation steps and covering 22.85% (16/70) of all isolates. Twenty-seven polymorphic fields were detected, 77.77% (21/27) of which were parsimony-informative, and secondary haplotype and nucleotide diversity were observed. Additionally, we detected high intraspecies haplotype diversity (hd: 0.933) and high nucleotide diversity (π: 0.00383), with 27 different nucleotide variation positions among the haplotypes of the isolates. Tajima's D value was negative, indicating population expansion and/or selection purification. The significantly negative Fu's Fs values indicated recent population expansion or the presence of expected rare haplotypes. CONCLUSION: The results of this study confirmed that C. tenuicollis isolates clustered in one lineage and were closely related to the relevant reference sequences in different countries, confirming the circulation of C. tenuicollis in different geographical regions.

Molecular characterization and haplotypes of hydatid cyst isolates collected from humans and ruminants in Setif Province (northeast of Algeria) based on mitochondrial cytochrome C oxidase subunit 1 (mt-CO1) gene sequences.

Cystic echinococcosis (CE) is a disease that can be transmitted from animals to humans, caused by the metacestode of Echinococcus granulosus. The disease has significant health and economic impacts worldwide, particularly in endemic areas. The study aimed to evaluate the prevalence of hydatid cysts in ruminants (cattle and sheep) (n = 2060) from the Setif Province of Algeria using microscopy. The results showed that hydatid cysts were detected in 9.6% (198/2060) of ruminants, with a higher prevalence in cattle (16.8%; 56/333) compared to sheep (8.2%; 142/1727). Molecular techniques were used to analyze a subset of animals consisting of 30 sheep and 4 cattle. Specifically, a fragment of the mitochondrial cytochrome c oxidase subunit 1 (mt-CO1) gene was sequenced and compared to sequences from seven humans from the same region. The results indicated that all isolates were identified as E. granulosus sensu stricto. Haplotype analysis identified 19 E. granulosus s.s. haplotypes arranged like a star, with the dominant haplotype (Hap04) at the center. Hap04 has been assigned a total of 17 positives, including positives from sheep, cattle, and two humans. This study is noteworthy for being the first to use a molecular approach to human and ruminant echinococcosis in Setif, a significant breeding region in Algeria.

Molecular discrimination of G1 and G3 genotypes of Echinococcus granulosus sensu stricto obtained from human, cattle, and sheep using the mitochondrial NADH dehydrogenase subunit 5 marker.

Cystic echinococcosis (CE) is a common zoonotic disease caused by the larval form of Echinococcus granulosus sensu lato. This study determined the genotype and haplotype differences using the NADH dehydrogenase subunit 5 gene in hydatid cyst samples. Human (n = 12), cattle (n = 28), and sheep (n = 31) hydatid cyst isolates were included. Seventy-one genomic DNA samples were successfully extracted, and a 759 bp mitochondrial NADH dehydrogenase subunit 5 gene fragment was amplified by PCR. Following the sequence analysis, E. granulosus sensu stricto isolates were identified as G1 (n = 61) and G3 (n = 10). A total of 23 haplotypes were obtained from the 71 E. granulosus s.s. G1 and G3 samples. The main haplotype was Hap01 (60.56 %), which consisted of the G1 genotype. The second largest haplotype was Hap04, which consisted entirely of the G3 genotype. Hap14 acted as a bridge between the G1 and G3 genotypes. This study identifies G1 as the dominant genotype in humans and farm animals in Turkey. High haplotype and nucleotide diversity in genotypes were observed. Additionally, this is the first report on the phylogeography and gene flow models of the E. granulosus s.s. population in Turkey using the NADH dehydrogenase subunit 5 gene, the best marker distinguishing between G1 and G3 genotypes.

Sequence and Haplotype Analyses of Ligula intestinalis in Acanthobrama marmid (Cyprinidae) in Turkey.

PURPOSE: Ligulosis caused by Ligula intestinalis adversely affects the fisheries carried out in the lakes and ponds, causing economic losses in the fish industry. In this study, it was aimed to reveal the molecular characterization of L. intestinalis isolates obtained from woodfish (Acanthobrama marmid) in Keban Dam Lake in Elazig province of Turkey by using mt-CO1 gene sequences and to determine the genetic differences and haplotypes between the isolates. METHODS: In the examination made in terms of L. intestinalis, the intestine of the fish was opened with the help of fine-tipped scissors, the contents were allowed to come out, and the parasites were taken into a petri dish containing phosphate buffered saline (PBS). Then, L. intestinalis plerocercoids were taken into 15 ml falcon tubes containing 70% ethanol and stored at - 20 °C until further analysis. From each isolate, total gDNA was extracted from the plerocercoids. A partial (480 bp) mt-CO1 gene was amplified by PCR and sequenced unidirectionally. The final size of the trimmed sequences was 392 bp for 43 sequences. Sequence and haplotype analyses were performed, followed by phylogenetic analyses. RESULTS: All isolates were confirmed as L. intestinalis by BLAST analysis. In addition, 87 nucleotide mutation positions were determined among 43 CO1 gene sequences. As a result of the haplotype network performed for the mt-CO1 gene region of L. intestinalis isolates; arranged in a star-like configuration with the main haplotype (Hap05), separated from other haplotypes by 1-6 mutation steps, and 29 haplotypes were identified, covering 13.9% (6/43) of the total isolates. Also, 75 variable (polymorphic) sites were determined, 52 of which were parsimony informative sites. CONCLUSIONS: The molecular characterization of L. intestinalis in woodfish (A. marmid) was identified for the first time in Turkey.

Successful management of delayed-onset adenosine deaminase deficiency with novel mutation.

A 4-year-old boy presented with acute-onset autoimmune cytopenia with severe, persistent lymphopenia, autoimmune thyroiditis, elevated IgE and glucose 6-phosphate dehydrogenase enzyme deficiency. In immunologic evaluation, lower T, B and natural killer cells and higher levels of adenosine deaminase (ADA) metabolites were observed. The compound heterozygous novel ADA gene mutations causing ADA deficiency were detected. Successful immunologic and metabolic cure was achieved with enzyme replacement therapy, followed by reduced intensity conditioning hematopoietic stem cell transplantation from a matched unrelated donor. An interesting aspect of this patient is the detection of novel compound heterozygous mutations without consanguinity and a secondary outcome is the recovery of glucose 6-phosphate dehydrogenase deficiency after hematopoietic stem cell transplantation.

Genetic diversity and haplotypes of Echinococcus granulosus isolated from cattle and buffaloes and first report of E. ortleppi (G5) in buffaloes in Pakistan based on mitochondrial cytochrome c oxidase subunit-1 gene (mt-CO1) markers.

Cystic echinococcosis (CE) is a parasitic disease that is caused by larval stage of Echinococcus granulosus tapeworm, one of the most important and neglected zoonotic disease. Although the echinococcosis is endemic in the neighboring countries, information regarding circulating genotypes of E. granulosus sensu lato is scarce in Pakistan. Therefore, the main purpose of this report was to contribute in molecular epidemiology and to find genetic variation and haplotypes of E. granulosus s.l. in cattle and buffalo isolates. To identify species circulating in country, parasite samples were collected from different slaughterhouses and butcher shops of two major cities, Rawalpindi and Peshawar located in Punjab and Khyber Pakhtunkhwa (KP) provinces, Pakistan, respectively. A total of 100 CE cyst samples were investigated from buffalo (n = 61), and cattle (n = 39) hosts. After genomic DNA extraction from individual cyst materials, mt-CO1 (875 bp) gene was amplified by PCR. After that, PCR products were electrophoresed on the agarose gel then purified and sequenced using forward primer. The sequences were trimmed (779 bp), aligned and matched with NCBI published sequences. E. granulosus s.s. (G1, G3) (71.4%; n = 20/28) was confirmed as the dominant species in buffalo and cattle. E. ortleppi (G5) (28.6%; n = 8/28) was recorded for the first time in both buffalo and cattle isolates from Rawalpindi. E. granulosus s.l. haplotype network showed single predominant haplotype, which comprised 40% of population. Tajima's D and Fu's Fs were negative and significant for E. ortleppi (G5), suggesting population expansion in Pakistan. Therefore, more studies using isolates of E. granulosus s.l. from various locations and intermediate hosts across Pakistan will add new data on molecular epidemiology and genotyping for effective control strategies of CE in Pakistan.

miRNA based biomarkers for the early diagnosis of Echinococcus granulosus in experimentally infected dogs.

Cystic echinococcosis, which is caused by the Echinococcus granulosus. Carnivores, as final hosts, contain adult tapeworms in the small intestine, and a variety of mammals, including humans, harbor the metacestod. This study was designed to investigate the miRNA-based biomarkers for early and accurate diagnosis of E. granulosus in experimentally infected dogs. A liver with an obvious hydatid cyst was obtained from a slaughterhouse and then protoscoleces were collected. Following, viable protoscoleces were administred to three experimental dogs (ED1, ED2 and ED3) and another uninfected control dog (UCD) was kept as control without infection. Stool samples of all groups were collected during 50 days from the beginning of the experimental infection and stored at - 80 °C till work. Total miRNA was isolated from all individual stool samples. The qRT-PCR method was used to determine the differences in the expression levels of E. granulosus specific miRNAs which were egr-let-7-5p, egr-miR-2b-5p, egr-miR-71-5p and egr-miR-125-5p. All miRNAs were found to be expressed from the first day in all infected dogs. In the stool samples of the UCD, the egr-miR-71-5p was detected, while the other miRNAs (egr-let-7-5p, egr-miR-2b-5p, egr-miR-125-5p) were not expressed. The expression of egr-let-7-5p and egr-miR-125-5p was significantly increased in ED1 compared to UCD on all days. In particular, for the first time, the expression levels of egr-let-7-5p and egr-miR-125-5p increased significantly between days 15 and 19. Similarly, the increase in let-7-5p and miR-125-5p expression was statistically significant in ED2. In ED3, egr-let-7-5p, egr-miR2b-5p and egr-miR-125-5p expressions were significantly increased on all days. In particular, egr-let-7-5p expression levels increased significantly for the first time between days 15 and 19. In addition, egr-mir-125-5p expression levels were found to increase at a high level for the first time on day 16. In conclusion, especially egr-let-7-5p and egr-miR-125-5p can be used as early diagnostic biomarkers in dogs infected with E. granulosus.

The importance of skin prick test in differentiating cat sensitivities and allergies in children.

BACKGROUND: The incidence of cat allergies in children has increased over the years. Children with cat allergies have mostly reported respiratory symptoms. The skin prick test (SPT) is the most preferred method to demonstrate sensitization to allergens. However, not all children who develop cat sensitization due to environmental exposure become allergic to cats. In our study, we aimed to determine the frequency of sensitization to cat and cat allergy, cat-related symptoms, and the cut-off value for the SPT that may indicate cat allergy. METHODS: Patients aged 2-18 years, who applied to the Health Sciences University Izmir Dr Behçet Uz Pediatrics and Surgery Training and Research Hospital and Balikesir University Application and Research Hospital Pediatric Allergy outpatient clinics between January 01, 2019 and December 31, 2020, were included in the study. Patients who underwent SPT and found to be sensitized to cat allergen, were evaluated retrospectively. Clinical and laboratory findings of the patients were recorded. Receiver operating characteristics (ROC) analysis was performed to determine the cut-off value for the SPT. RESULTS: Sensitization to cat was detected in 140 (4%) out of 3499 patients who underwent SPT. The median age of the patients was 12 years (min-max: 5-18) and 67.1% were male. Eighty-eight (62.9%) patients were symptomatic upon contact with cats, predominantly with nasal symptoms. These patients had significantly larger cat SPT wheal size than asymptomatic patients. The cut-off value was determined as 5.5 mm with a sensitivity of 72.7% and a specificity of 61.5% (95% CI, 60.5%-78.4%). Symptoms resolved in about half of our patients by reducing contact with cats. DISCUSSION: The present study is the first to report the frequency and clinical findings of cat sensitizations and allergies in Turkish children. For effective treatment, cat allergy must be diagnosed. In this regard, the use of a practical, readily accessible 5.5 mm cut-off point on the SPT may be helpful.

Molecular characterization of cattle and sheep isolates of Echinococcus granulosus from Elazig province in Turkey and expression analysis of the non-coding RNAs, egr-miR-7, egr-miR-71 and egr-miR-96.

Cystic Echinococcosis (CE) is a common zoonotic disease seen in human and animals worldwide, caused by the larval form of Echinococcus granulosus. In this study, E. granulosus s.l. species and haplotypes were determined in hydatid cysts isolated from cattle and sheep, and the expression levels of egr-miR-7, egr-miR-71 and egr-miR-96 miRNAs were compared in different cyst structures. A total of 82 (cattle, n = 41; sheep, n = 41) hydatid cyst isolates (germinal membranes and/or protoscoleces) were collected from a slaughterhouse in Elazig province of Turkey. After mt-CO1 gene sequences were made, 81 out of 82 hydatid cyst isolates were determined as E. granulosus s.s. (G1 and G3), while an isolate of cattle origin was determined as Echinococcus canadensis (G6/7). A total of 26 nucleotide polymorphisms and 29 haplotype groups were identified in the samples. miRNA expressions in germinal membranes of sterile cysts and germinal membrane and protoscoleces of fertile cysts were investigated by qRT-PCR and Real Time PCR analyses. It was determined that miRNAs were expressed at high levels in 79.31% of the 29 haplotype groups and at low levels in the remaining 10.34%. In 10 fertile samples of sheep origin, egr-miR-7, egr-miR-71 and egr-miR-96 miRNAs were found to be 44, 168, and 351-fold higher in expression, respectively, in the germinal membrane compared to the protoscoleces. Especially egr-miR-96 may have the potential to be used as biomarkers in the diagnosis of active CE.

Co-infection of Echinococcus equinus and Echinococcus canadensis (G6/7) in a gray wolf in Turkey: First report and genetic variability of the isolates.

Cystic echinococcosis (CE) is an important zoonotic diseases caused by larval form of Echinococcus granulosus sensu lato. The material of this study was the gray wolf (Canis lupus), which was found dead in the rural area of Bingol province of Turkey. The animal was brought to Veterinary Faculty for necropsy and many of adult Echinococcus spp. obtained. A total of 9 whole adult worms were morphologically examined under the microscope, gDNA was isolated from individual samples, a partial mt-CO1 gene fragment (875 bp) was amplified with PCR and sequenced. According to the phylogenetic analysis, six worms were characterized as E. equinus, while three were reported as E. canadensis (G6/7). It was found that the haplotypes of both species were similar to previously published haplotypes. This is the first report in which E. equinus and E. canadensis (G6/7) adult parasites were detected together in a gray wolf's intestine. The findings are important in that it draws attention to the importance of wild cycle in the spread of CE.

In Silico Evaluation of the Haplotype Diversity, Phylogenetic Variation and Population Structure of Human E. granulosus sensu stricto (G1 Genotype) Sequences.

Echinococcus granulosus sensu lato is the causative agent of cystic echinococcosis (CE), which is a neglected zoonotic disease with an important role in human morbidity. In this study, we aimed to investigate the haplotype diversity, genetic variation, population structure and phylogeny of human E. granulosus sensu stricto (s.s.) (G1 genotype) isolates submitted to GenBank from different parts of the world by sequencing the mitochondrial CO1 and ND1 genes. The sequences of the mt-CO1 (401 bp; n = 133) and mt-ND1 (407 bp; n = 140) genes were used to analyze the haplotype, polymorphism and phylogenetic of 273 E. granulosus s.s. (G1 genotype) isolates. Mutations were observed at 31 different points in the mt-CO1 gene sequences and at 100 different points in the mt-ND1 gene sequences. Furthermore, 34 haplotypes of the mt-CO1 sequences and 37 haplotypes of the mt-ND1 sequences were identified. Tajima's D, Fu's Fs, and Fu's LD values showed high negative values in both mt-CO1 and mt-ND1 gene fragments. The haplotype diversities in the sequences retrieved from GenBank in this study indicate that the genetic variation in human isolates of E. granulosus s.s. in western countries is higher than in eastern countries. This may be due to demographic expansions due to animal trades and natural selections.

Genetic Diversity and Haplotype Analysis of Cattle Hydatid Cyst Isolates Using Mitochondrial Markers in Turkey.

Echinococcus granulosus sensu lato (s.l.) causes cystic echinococcosis in ungulates and humans. The current study was designed to find the genetic diversity and haplotypic profiles of hydatid cysts from the lungs of cattle in three provinces in eastern Turkey. Individual cyst isolates (n = 60) were collected from infected cattle lungs after slaughter and then samples were stored in ethanol (70%) until further use. From each isolate, total gDNA was extracted from the cysts' germinal layers. A partial (875 bp) mt-CO1 gene was amplified by PCR and sequenced unidirectionally. The final size of the trimmed sequences was 530 bp for 60 sequences. Sequence and haplotype analyses were performed, followed by phylogenetic analyses. According to BLAST searches, all sequences were detected as E. granulosus s.s. (G1 and G3 strains). Forty-nine point mutations were identified. In addition, five conserved fragments were detected in all sequences. The haplotype analysis diagram showed E. granulosus s.s. haplotypes organized in a star-like configuration. The haplotypes were characterized by 1-17 mutations compared with the fundamental focal haplotype. Thirty-three haplotypes were determined in 60 samples of which 17 (28.3%) belonged to the main haplotype (Hap_06). The mt-CO1 sequences revealed 49 polymorphic sites, 34.5% (20/49) of which were informative according to parsimony analysis.

Parasite and Cancer Relationship

Cancer is a life-threatening disease that occurs as a result of the uncontrolled proliferation of cells in any organ or tissue of the body. Parasites are dangerous organisms that can cause death in some cases. Parasite and cancer cells are similar in their capacity to survive and proliferate independently of exogenous growth factors, to be resistant to apoptosis, and to evade host immune mechanisms. Therefore, it is difficult for the body to completely get rid of cancer cells and parasitic agents. In vitro studies or experimental animal studies examining the parasite-cancer relationship have shown that besides parasites that can cause cancer directly, there are also parasites that can indirectly stimulate cancer development through various mechanisms. On the other hand, it is known that the immune response against some parasites can show antitumoral activity in the body. Parasitic agents can have both tumoral and antitumoral effects through regulation of immune response, prevention of metastasis and angiogenesis, inhibition of proliferative signals, and regulation of inflammatory responses that induce cancer development.

Prevalence of Viruses in Acute Asthma Exacerbations in Childhood in a Hospital in West Part of Turkey.

Acute asthma exacerbations (AAE) are episodes characterized by potentially life-threatening and rapidly deteriorating asthma symptoms. Viral respiratory infections are one of the major triggers in the pathophysiology of childhood asthma exacerbations. In this study, we aimed to determine the distribution of viral agents among pediatric AAE patients. One hundred and three AAE patients, aged 5 or older, hospitalized between from February 2017 through February 2020 at Pediatric Immunology and Allergic Diseases Unit were included in this study. Fifty patients (48.5%) were female, and the mean age of the patients was 108.2 months. Viruses were detected in 58 (%56.3) of the patients, in 5 of whom more than one virus type was detected. The most commonly detected virus was human rhinovirus (n=43, 67.1%). Other types included respiratory syncytial virus (n=8; 12.5%), influenza (n=6; 9.3%), human metapneumovirus (n=4; 6.2%), adenovirus (n=1; 1.5%), enterovirus (n=1; 1.5%), and parainfluenza (n=1; 1.5%). Viral agents were detected in 29 out of the 47 patients with allergic asthma, with human rhinoviruses comprising the majority (18 patients). The mean length of hospital stay was 7.89 days. Human rhinovirus is the most common virus that triggers AAE, with similar distributions in allergic and non-allergic asthma. We found no correlation between virus type and the length of hospital stay.

Cloning, expression and serodiagnostic potential of HSP70 of Taenia multiceps in sheep.

The aim of this study was to determine the efficacy of the recombinant protein expressed by the T. multiceps HSP70 gene in the immunodiagnosis of sheep coenurosis. Specific primers were designed to amplify the gene encoding HSP70 from the whole genome sequence of T. multiceps, mRNA was isolated from C. cerebralis cysts obtained from an infected sheep's brain, and then, cDNA was isolated. The gene region related to the designed primers was amplified by PCR, and the obtained product was cloned and expressed. The specificity of the recombinant protein purified afterwards was demonstrated by Western Blot using known positive and negative sheep sera. Additionally, the efficiency of the recombinant protein in ELISA was determined with sera collected from 1207 sheep. Finally, the sensitivity and specificity values ​of the recombinant TmHSP70 antigen in the Western Blot were 100 %, while the sensitivity and specificity rates of the ELISA were 66.6 % and 52.6 %, respectively. At the same time, 567 (46.97 %) of the 1207 analyzed serum samples were found to be positive by ELISA. In conclusion, the Western Blot technique had a higher quality than ELISA in detecting the TmHSP70 antigen for the serodiagnosis of sheep coenurosis.

Molecular Characterization and Haplotype Analyses of Lung Hydatid Cyst Isolates of Cattle and First Report of Echinococcus canadensis (G6/G7) in Cattle Isolates in Turkey.

PURPOSE: Cystic echinococcosis (CE) is a zoonotic disease, caused by parasite known as Echinococcus granulosus sensu lato complex, leading to substantial economic losses in rural areas with public health problems. This study was carried out to understand the haplotypic profiles of the partial mitochondrial cytochrome c oxidase (mt-CO1) gene of cattle lung hydatid cyst samples which were obtained from two provinces in Turkey. METHODS: In this study, forty (n = 40) hydatid cyst samples from the lungs of slaughtered cattle were obtained. The germinal layers were taken separately for each individual cyst then stored in 70% ethanol. From each individual cyst sample, total genomic DNA was extracted. Amplification of the partial mt-CO1 gene (875 bp) was performed using a specific primer set by PCR, and then, the amplicons were sequenced. All sequences were analyzed individually, followed by alignment, and haplotype and phylogenetic analyses were then performed. RESULTS: By the sequence alignment process, 39 out of the 40 sequences were characterized as E. granulosus sensu stricto. However, one of them was matched with E. canadensis (G6/G7). The haplotype analyses of the E. granulosus s.s. isolates were arranged in a star-like orientation with a main haplotype, which was separated from other haplotypes by 1-10 mutation steps, and 26 haplotypes were identified. In the mt-CO1 sequences, 29 polymorphic sites were determined, and 34.5% (10/29) of them were parsimony informative. CONCLUSIONS: The findings of this study provide the first report of E. canadensis (G6/G7 genotype) among cattle in Turkey.

Genetic diversity and haplotypes of paediatric hydatid cyst isolates and first occurrence of E. canadensis (G6/G7) in paediatric cases in Turkey.

Cystic echinococcosis (CE) is a neglected zoonotic tropical disease caused by Echinococcus granulosus sensu lato. The aim of this study was to investigate the genetic variation of hydatid cyst isolates obtained from surgically confirmed paediatric cases originating from two different regions in eastern Turkey. Seventeen paediatric cases aged between 6 and 16 were operated by open surgery, and the germinal layers of their cysts were obtained for further molecular analyses. After genomic DNA isolation, 875 bp mt-CO1 gene fragments were amplified in all samples by PCR. Then, the unidirectional sequence analyses of the PCR products were carried out. According to the BLAST analyses of 17 sequences, 16 of these sequences were matched with E. granulosus sensu stricto, while one sequence was identified as E. canadensis (G6/G7) for the first time in paediatric cases in Turkey. High haplotype diversity and low nucleotide diversity were observed in the E. granulosus s.s. sequences.

Haplotype comparisons of Echinococcus granulosus sensu lato via mitochondrial gene sequences (co1, cytb, nadh1) among Pakistan and its neighbouring countries.

Echinococcus granulosus sensu lato (s.l.) is a zoonotic parasite that causes cystic echinococcosis (CE) in humans. However, E. granulosus sensu stricto (s.s.) is considered the predominant species in CE infections worldwide. According to the population genetic diversity and structure of E. granulosus s.l., gene flow can explain the parasite drift among the neighbouring countries of Pakistan. The mitochondrial (mt) co1 (n = 47), nadh1 (n = 37) and cytb (n = 35) nucleotide sequences of E. granulosus s.l. isolates from Pakistan, Iran, China and India were retrieved from the National Centre for Biotechnology Information database to determine the genealogical relationships. The sequences were grouped as the mt-co1 (genotypes G1 and G3, G6-G7), mt-cytb (genotypes G1 and G3), and mt-nadh1(genotypes G1 and G3). The data were analysed using bioinformatic tools. A total of 19 polymorphic sites for the mt-co1 sequence (374 bp) were observed of which 31.6% (6/19) were parsimony-informative sites. Unique singleton haplotypes within the E. granulosus s.s. haplotype network based on the mt-co1 gene were highly prevalent (68.4%; 13/19) in Pakistani isolates followed by Chinese, Indian and Iranian isolates; four polymorphic sites were detected in the E. canadensis (G6/G7). In E. canadensis mt-co1 haplotype network, 75% (3/4) unique singleton haplotypes were from the Iranian isolates. Twelve polymorphic sites were found using the mt-cytb sequence (547 bp); 25% (3/12) were parsimony-informative and there were 66.7% (8/12) unique singleton haplotypes within the mt-cytb haplotype network in E. granulosus s.s. with the most reported from Pakistan followed by Iran and China. 20 polymorphic sites were detected in E. granulosus s.s. mt-nadh1 sequences (743 bp); 20% (4/20) were parsimony-informative. There were 66.7% (8/12) main single haplotypes within the mt-nadh1 haplotype network, with the most reported from Pakistan followed by that from India, Iran and China. The sequence analyses show low nucleotide diversity and high haplotype diversity in general.

Molecular Characterization of Hydatid Cysts Cases in a Wild Boar and Mule in Turkey

Objective: Cystic echinococcosis (CE) is a zoonotic infection that affects humans, livestock and wild animals through the larval form of Echinococcus granulosus sensu lato (s.l.). Molecular and taxonomic studies carried out in the recent years accept that Echinococcus granulosus s.l., a complex of 5 cryptic species, causes CE. In this study, we performed morphological and molecular characterisation of cyst isolates obtained from a wild boar and mule naturally infected with hydatid cyst. Methods: After gDNA isolation, the mitochondrial cytochrome oxidase subunit 1 (mt-CO1) gene region was amplified by polymerase chain reaction (PCR) with specific primers. The amplified mt-CO1 PCR products were purified and one-way DNA sequence analysis was performed. Results: Comparison of the partial sequences of mt-CO1 gene from the hydatid cyst isolates with that of reference sequences in GenBank revealed 100% similarity with E. granulosus sensu stricto (G1-G3) sequences. Conclusion: To the best of our knowledge, this is the first study to determine the molecular characterisation of Echinococcus species in a wild boar in Turkey.

A Case-Study of the Molecular Diagnosis of Echinococcus multilocularis in Wild Boar with Comments on its Public Health Significance in Turkey.

Echinococcus multilocularis is a parasite species of zoonotic importance which can be fatal to humans and causes Alveolar Echinococcosis (AE). This report describes the development of a cyst from the liver of a wild boar and molecular confirmation of its identification. The cyst material was obtained from the liver of a wild boar killed by hunters. Genomic DNA was extracted from the germinal layer of the cyst material, and 875 bp mitochondrial cytochrome c oxidase subunit I (COI) gene fragment was amplified by PCR and sequenced. A BLAST search matched 100% with published Echinococcus multilocularis sequences. This study confirms the occurrence of E. multilocularis in a wild boar for the first time in Turkey.