Molecular Study of Cysticercus Tenuicollis from Slaughtered Sheep in Sulaymaniyah Province, Iraq.
ABSTRACT: Introduction:Cysticercosis caused by the larval stage of Taenia hydatigena is economically the most important endemic parasitic disease in Iraq. Few data are available relating to the genetic divergence of this helminth. This study aimed to molecularly characterise Cysticercus tenuicollis isolates from sheep in Sulaymaniyah province, Iraq. Material and Methods:DNA extraction and amplification of specimens of C. tenuicollis from 46 sheep were conducted by PCR for the mitochondrial 12S rRNA gene. The 19 amplicons were subjected to purification and partial sequencing. Results:Five 12S rRNA nucleotide sequence haplotypes were found. The pairwise nucleotide difference between haplotypes of 12S rRNA gene ranged from 0.2% to 0.7%. Four out of the five haplotypes of C. tenuicollis contained one to two base mutations and were discovered in Iraq for the first time, and this may be a unique mutation globally which has not been recorded previously. Three newly recorded haplotypes contained only one single mutation, and the other one contained two mutations. Phylogenetic analysis showed that all isolated strains were closely related to Iranian sheep isolates. Conclusions:Four new strains of T. hydatigena were discovered for the first time in the study area.
Project description:Taenia multiceps and Taenia hydatigena are widely distributed tapeworms of canids. Due to a lack of genetic information on these two parasites in China, in this study we analyzed six coenurus cerebralis and two cysticercus tenuicollis cysts from goats or sheep in Inner Mongolia, northern China by amplifying three mitochondrial genes (cox1, nad4, and cytb). Two haplotypes were obtained at each locus for either of the two Taenia cestode species, with ten nucleotide sequences being novel. The degrees of genetic variations were 1.18%, 0.61% and 0.52% for coenurus cerebralis, and 0.24%, 0.46% and 0.35% for cysticercus tenuicollis at the cox1, nad4 and cytb loci, respectively. This is the first molecular description of animal-derived metacestodes of T. multiceps and T. hydatigena in Inner Mongolia, China. Novel nucleotide sequences might reflect endemic genetic characterization of the two cestodes. The present data are useful to explore the biological and epidemiological significance of intra-specific variations within both Taenia cestodes.
Project description:Cysticercus tenuicollis, commonly known as "water bell," is a larva of Taenia hydatigena, which is the most significant parasite of pigs. However, until now very few information is available regarding the prevalence and genetic characterization of the Cysticercus tenuicollis in Tibetan pigs. Therefore, the aim of this study was to investigate the prevalence and phylogenetic analysis of Cysticercus tenuicollis in Tibetan pigs. For this purpose, the COX2 gene of Cysticercus tenuicollis was amplified and sequenced for the first time in Tibetan pigs. The overall prevalence of Cysticercus tenuicollis was 43.93% in Tibetan pigs, with further distribution of 42.86% in 2014 and 45.35% in 2015. In Tibetan male and female pigs, the prevalence of Cysticercus tenuicollis was 43.39% and 44.56%, respectively. The prevalence of Cysticercus tenuicollis in different growing stages (juveniles, subadults, and adults) varied from 30.20% to 63.79%. The phylogenetic analysis of the Cysticercus tenuicollis isolates showed very close resemblance to 16 reference strains, isolates from Gansu, Hunan, and Sichuan provinces of China. To the best of our knowledge, this is the first report on the prevalence and genetic characterization of Cysticercus tenuicollis derived from Tibetan pigs. The data of present study provides baseline information for controlling cysticerci infections in pigs in Tibetan Plateau, China.
Project description:The cestode Taenia hydatigena uses canids, primarily dogs, as definitive hosts, while the metacestode larval stage cysticercus infects a range of intermediate hosts, including domestic animals such as goats, sheep, and pigs. Cysticercosis due to T. hydatigena has large veterinary and economic drawbacks. Like other taeniids, e.g., Echinococcus, intraspecific variation is found among the members of the genus Taenia. In Africa, few studies are available on the epidemiology and distribution of T. hydatigena, and even fewer studies are available on its genetic variation. In this study, we molecularly identified 11 cysticerci from sheep in Sudan and demonstrated the genetic variation based on the NADH dehydrogenase subunit 1 (nad1) and cytochrome c oxidase subunit 1 (cox1) mitochondrial genes. The isolates were correctly identified as T. hydatigena with more than 99% similarity to those in the GenBank database. Low diversity indices and insignificant neutrality indices were observed, with 3 and 2 haplotypes for the nad1 and cox1 genes, respectively. The results suggest the presence of unique T. hydatigena haplotypes in Sudan, as haplotypes with 100% similarity were not found in the GenBank database. With few available studies on the genetic variation of T. hydatigena in Africa, this report represents the first insights into the genetic variation of T. hydatigena in Sudan and constitutes useful data.
Project description:BACKGROUND:Cysticercosis caused by cysticercus tenuicollis is a metacestode infection that affects several species of ungulates. It is caused by the larval stage of Taenia hydatigena, an intestinal tapeworm in dogs and wild canids. In the intermediate host, the mature cysticerci are usually found in the omentum, mesentery, and peritoneum, and less frequently in the pleura and pericardium. The migrating larvae can be found mostly in the liver parenchyma causing traumatic hepatitis in young animals. Most infections are chronic and asymptomatic, and are diagnosed at the abattoir. The acute form of infection is unusual in sheep and reports of death in lambs are rare. METHODS:In March 2018, fifteen female lambs presented anorexia, weakness, lethargy, and death secondary to acute visceral cysticercosis. Twelve of them underwent hepatic ultrasonography. Examinations were performed on standing or left lateral recumbent animals. RESULTS:Livers of affected animals presented rounded margins and a thickened, irregular and hyperechoic surface. Hepatic parenchyma appeared to be wholly or partially affected by lesions characterized by heterogeneous areas crossed by numerous, irregular, anechoic tracts ranging from 1 to 2 cm in length and 0.1 to 0.2 cm in width. Superficial and intraparenchymal cystic structures were also visualized. The presence of lesions was confirmed by anatomopathological examination, and T. hydatigena cysticerci was identified by morphological and molecular characterization of isolates. CONCLUSIONS:Our results highlighted that hepatic ultrasonography is effective for an intra-vitam diagnosis of acute cysticercosis in lambs.
Project description:Several developments have been recently achieved to understand pet-dog parasites and their relationship with hosts; however, parasites' presence and distribution in shepherd-dog have been mainly neglected; this knowledge gap is of critical sanitary importance, as shepherd-dogs could harbor zoonotic helminths including Echinococcus granulosus sensu lato. The related human disease, cystic echinococcosis, is a worldwide neglected disease, with high endemicity in the Mediterranean Basin. To evaluate the presence of E. granulosus and other parasites, a sheep-dog population from the province of Grosseto (Tuscany, Italy) has been investigated. Overall, 648 dog fecal samples obtained from 50 modern sheep farms, having a total of 216 dogs, were collected. Specimens were analyzed using a standardized centrifugal flotation method (specific gravity = 1.3). Taeniid eggs detected were further isolated using a sieving/flotation technique. DNA was isolated from eggs for PCR and sequence analyses for species identification (gene target: 12S rRNA and nad1). Thirty-nine (78%) farms tested positive for at least one parasite species or genus. The most represented intestinal helminths were Toxocara spp. in 64% of farms, followed by Ancylostomatidae (58%), Trichuris vulpis (50%), Capillaria spp. (34%), and taeniids (32%). Sequence analyses confirmed the presence of Taenia hydatigena in seven farms, Taenia (syn. Multiceps) multiceps in five farms, and T. pisiformis in one farm. No DNA was extracted from four previously taeniid egg-positive farms. No amplification of amplicon corresponding to E. granulosus was achieved in the investigated farms. Although not entirely expected, Spearman's test showed a positive correlation between flock size and the number of dogs per farm (? = 0.588, P < 0.001). The quantitative analysis reported that the home slaughter practice was affected neither by the flock size nor by the number of dogs per farm. The probability to diagnose farms positive for taeniids had been increased by about 35% for each dog unit increase [odds ratio (OR) = 1.35, P = 0.012]. In conclusion, the wide distribution of T. hydatigena and T. multiceps detected in the present study clearly reveals that dogs have still access to raw offal, a major risk for the transmission of E. granulosus. Home slaughtering is an unavoidable practice, and more efforts must be undertaken by the public health system to prevent and control potential zoonotic taeniids.
Project description:BACKGROUND:Cysticercosis caused by the metacestode larval stage of Taenia hydatigena is a disease of veterinary and economic importance. A considerable level of genetic variation among isolates of different intermediate hosts and locations has been documented. Generally, data on the genetic population structure of T. hydatigena is scanty and lacking in Nigeria. Meanwhile, similar findings in other cestodes like Echinococcus spp. have been found to be of epidemiological importance. Our aim, therefore, was to characterize and compare the genetic diversity of T. hydatigena population in Nigeria based on three mitochondrial DNA markers as well as to assess the phylogenetic relationship with populations from other geographical regions. METHODS:In the present study, we described the genetic variation and diversity of T. hydatigena isolates from Nigerian sheep and goats using three full-length mitochondrial genes: the cytochrome c oxidase subunit 1 (cox1), NADH dehydrogenase subunit 1 (nad1), and NADH dehydrogenase subunit 5 (nad5). RESULTS:The median-joining network of concatenated cox1-nad1-nad5 sequences indicated that T. hydatigena metacestodes of sheep origin were genetically distinct from those obtained in goats and this was supported by high FST values of nad1, cox1, and concatenated cox1-nad1-nad5 sequences. Genetic variation was also found to be higher in isolates from goats than from sheep. CONCLUSIONS:To the best of our knowledge, the present study described the genetic variation of T. hydatigena population for the first time in Nigeria using full-length mitochondrial genes and suggests the existence of host-specific variants. The population indices of the different DNA markers suggest that analysis of long mitochondrial DNA fragments may provide more information on the molecular ecology of T. hydatigena. We recommend that future studies employ long mitochondrial DNA sequence in order to provide reliable data that would explain the extent of genetic variation in different hosts/locations and the biological and epidemiological significance.
Project description:Cystic echinococcosis (CE) caused by E. granulosus is a serious helminthic zoonosis in humans, livestock and wildlife. Xinjiang is one of high endemic province for CE in China. A total of 55 sheep and cattle livers containing echinococcal cysts were collected from slaughterhouses in Changji and Yining City, northern region of Xinjiang. PCR was employed for cloning 2 gene fragments, 12S rRNA and CO1 for analysis of phylogenetic diversity of E. granulosus. The results showed that all the samples collected were identified as G1 genotype of E. granulosus. Interestingly, YL5 and CJ75 strains were the older branches compared to those strains from France, Argentina, Australia. CO1 gene fragment showed 20 new genotype haploids and 5 new genotype haplogroups (H1-H5) by the analysis of Network 5.0 software, and the YLY17 strain was identified as the most ancestral haplotype. The major haplotypes, such as CJ75 and YL5 strains, showed identical to the isolates from Middle East. The international and domestic trade of livestock might contribute to the dispersal of different haplotypes for E. granulosus evolution.
Project description:BACKGROUND:In farm animals, mitochondrial DNA (mtDNA) effect on economic performance remains hot-topic for breeding and genetic selection. Here, 53 maternal lineages of Small-tailed Han sheep were used to investigate the association of mitochondrial DNA variations and the lambing litter size. RESULTS:Sequence sweeping of the mitochondrial coding regions discovered 31 non-synonymous mutations, and the association study revealed that T7719G in mtDNA tRNA-Lys gene was associated with litter size (P?<?0.05), manifesting 0.29 lambs per litter between the G and T carriers. Furthermore, using the mixed linear model, we assayed the potential association of the ovine litter size and haplogroups and multiple-level mtDNA haplotypes, including general haplotypes, assembled haplotypes of electron transport chain contained sequences (H-ETC), mitochondrial respiratory complex contained sequences (H-MRC) and mitochondrial genes (H-gene, including polypeptide-coding genes, rRNA genes and tRNA genes). The strategy for assembled mitochondrial haplotypes was proposed for the first time in mtDNA association analyses on economic traits, although none of the significant relations could be concluded (P?>?0.05). In addition, the nuclear major gene BMPR1B was significantly correlated with litter size in the flock (P?<?0.05), however, did not interact with mtDNA T7719G mutation (P?>?0.05). CONCLUSIONS:Our results highlight mutations of ovine mitochondrial coding genes, suggesting T7719G in tRNA-Lys gene be a potentially useful marker for selection of sheep litter size.
Project description:Cyclophyllidean tapeworms obligatorily parasitize numerous mammalian species, including herbivores, domestic animals and humans, of which, the genera Taenia and Mesocestoides are well characterized. However, little is known about these parasitic infections in wild animals. This study aims to investigate the prevalence and distribution of Taenia sp. and Mesocestoides sp. in wild carnivores in Mongolia by identifying tapeworm species based on mtDNA analysis. The field survey was carried out in 2012-2013 in 19 provinces located in different ecological regions. A total of 405 fecal samples from wild carnivores were collected. Specific DNA markers in fecal samples was detected via copro-DNA analysis and tapeworm species were identified by DNA sequencing. From 27.7% (112/405) of samples, cox1 and 12S rRNA genes of tapeworms were amplified. Further, Taenia hydatigena (50.0%, 56/112) and two Mesocestoides species, including Mesocestoides sp.-1 (36.6%, 41/112) and Mesocestoides sp.-2 (13.4%, 15/112) were identified by DNA sequencing. The prevalence of T. hydatigena was 19.9% (27/136), 13.8% (23/167), 4.8% (3/62), and 7.5% (3/40) in wolves, red foxes, corsac foxes, and snow leopards, respectively. The prevalence of Mesocestoides sp.-1 was 14.7% (20/136), 9% (15/167), 9.7% (6/62) in wolves, red foxes, and corsac foxes, while the prevalence of Mesocestoides sp.-2 was 4.4% (6/136), 1.8% (3/167), 3.2% (2/62), and 10.0% (4/40) in wolves, red foxes, corsac foxes, and snow leopards, respectively. T. hydatigena was found throughout all ecological regions, while Mesocestoides sp.-1 was in the mountain taiga, forest-steppe, steppe, desert-steppe, and desert, and Mesocestoides sp.-2 in the alpine, forest-steppe, steppe, and desert-steppe ecoregions. This study revealed the prevalence and distribution of cyclophyllidean tapeworms in wild carnivores in Mongolia; while also confirming that wolves, red foxes, corsac foxes, and snow leopards serve as definitive hosts for unidentified Mesocestoides species.
Project description:This dataset investigates the transcriptional effect of mitochondrial 12S rRNA hypermethylation, both by overexpressing the mitochondrial methyltransferase mtTFB1 in HeLa cells and by using A1555G cybrids, where the 12S rRNA is hypermethylated. HeLa cells overexpressing a methyltransferase-deficient mtTFB1 (mtTFB1[G65A]) and wild-type A1555A cybrids were used as controls. four samples with 12S rRNA hypermethylation (two cell lines, with two biological replicates each) versus four samples with basal 12S rRNA methylation (two cell lines, with two biological replicates each)