Novel isolate of Cysticercus tenuicollis-Kalar isolate has been revealed in Kalar, Iraq

Introduction: Although Cysticercus tenuicollis is one of the most economic and veterinary important parasite in Iraq, scanty molecular characterization exists for this helminth. This study aimed to determine the prevalence and molecular description of C. tenuicollis isolates from sheep in Kalar district of Iraq. Methodology: A total of 2,906 slaughtered sheep were examined post-mortem. Up to 20 samples of C. tenuicollis was extracted and amplified using mitochondrial COX1 gene. Results: The overall prevalence rate was 6.88%, and female sheep recorded higher rate of infection (24.35%) than male (6.16%) with significant difference (p<0.05). The molecular results showed 14 haplotypes for COX1 gene and the pairwise nucleotide variation among them was ranged from 0.2 to 2.6%. Twelve out of fourteen haplotypes of C. tenuicollis involving one to three base mutations were discovered in Kalar, Iraq for the first time and this could be a unique mutation internationally and did not registered previously. Eleven newly recorded haplotypes involved only one single mutation and the remaining one involved three mutations. Phylogenetic interpretation showed that Cysticercus tenuicollis-Kalar isolate were clustered in one clade, and closely related to isolates discovered in Nigeria, China, Turkey, Poland, and Iran. Conclusions: This study provided a new record data on prevalence and discovered novel strains of C. tenuicollis in the study area for the first time named Cysticercus tenuicollis-Kalar isolate. Novel haplotypes might consider endemic genetic characterization of this metacestode. The present data may be useful to provide a good molecular background for future preventive and control programs.

Parasitic cysts in wild boars hunted in Central Italy: The sanitary controls in the wild game meats chain

Seven hundred sixty-five (765) adult wild boars were examined during the 2016/2017 hunting season for the research of parasites. Post mortem inspection was carried out at the slaughterhouse by the Official Veterinarian on the plucks (heart, tongue, lungs, diaphragm, and liver) of the killed animals presented by hunters. Of these, 0.8% (6/765) were positive for Echinococcus granulosus sensu lato (s.l.), and 2.6% (20/765) were positive for the metacestode stage of Taenia hydatigena (Cysticercus tenuicollis), while 0.5% (4/765) animals showed a mixed infection (Echinococcus granulosus s.l. and Taenia hydatigena). Sixty-three (63) cystic lesions were found. Of these 25,4% (16/63) were caused by Echinococcus granulosus s.l. and 74,6% (47/63) were caused by Cysticercus tenuicollis. The more involved organs were liver and lungs, in a less extension omentum and diaphragm. Parasitological analyses showed an overall prevalence of 3.9% for metacestodes in the hunted animals examined (Paoletti et al., 2018). Hydatids were molecularly characterized as E. granulosus sensu stricto. Trichinella spp. examination results showed no evidences of parasitic cysts. The products of hunting used for own consumption and direct sale to the final consumer or retailer, according to Regulation (EC) No 853/2004, lack of overall control by the Competent Authority. This is a critical point in the food chain of the game meat. The data obtained show the importance of the post mortem inspection and the central role of the Competent Authority to ensure not only the food safety of game meat but also to collect data for extensive epidemiological investigations on live-stocks wildlife settings having a direct impact on public health.

In-silico analysis of mt-CO1 gene of Taenia hydatigena sheep isolates

Summary Taenia hydatigena is a tapeworm that affects herbivores in different regions of the world. Cysticercus tenuicollis (larvae of T. hydatigena), is prevalent in ruminants and pigs. In the current study, phylogenetic analysis of the published mt-CO1 gene of C. tenuicollis sheep isolates was analyzed using in-silico method and vertical and horizontal transmission at the global level by using a meta-analysis approach. A total of 82 mt-CO1 nucleotide sequences (339 bp) of C. tenuicollis sheep isolates from the NCBI database (Italy -Sardinia-, Iran, Palestine, Iraq, Finland, India and China) were used to investigate haplotype and genetic relationships. Tajima’s D (-2,2984) value was negative for the mt-CO1 sequences signifying the population expansion and/or purifying selection. The highly negative Fu’s Fs (-60,528) values determined for the sequences reflecting the existence of uncommon haplotypes. The mt-CO1 of C. tenuicollis haplotype network had 47 haplotypes arranged within a star-like configuration with a main haplotype, which encompassed 25.6 % of the total isolates. In the mt-CO1 haplotype network analyzed, there were 80.5 % unique single haplotype and highest ratio was observed in C. tenuicollis from sheep originating from Iran, followed by Sardinia, Palestine and Finland. If the current condition continues, genetic differences in T. hydatigena will be able to rise, and possible new strains and/or genotypes that may influence the host adaptation and life cycle of the parasite may emerge.

Αcute visceral cysticercosis caused by Taenia hydatigena in lambs: ultrasonographic findings

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.

A new molecular nomenclature for Taenia hydatigena: mitochondrial DNA sequences reveal sufficient diversity suggesting the assignment of major haplotype divisions

Cysticercosis caused by the metacestode larval stage of Taenia hydatigena formerly referred to as Cysticercus tenuicollis is a disease of veterinary importance that constitutes a significant threat to livestock production worldwide, especially in endemic regions due to condemnation of visceral organs and mortality rate of infected young animals. While the genetic diversity among parasites is found to be potentially useful in many areas of research including molecular diagnostics, epidemiology and control, that of T. hydatigena across the globe remains poorly understood. In this study, analysis of the mitochondrial DNA (mtDNA) of adult worms and larval stages of T. hydatigena isolated from dogs, sheep and a wild boar in China showed that the population structure consists of two major haplogroups with very high nucleotide substitutions involving synonymous and non-synonymous changes. Compared with other cestodes such as Echinococcus spp., the genetic variation observed between the haplogroups is sufficient for the assignment of major haplotype or genotype division as both groups showed a total of 166 point-mutation differences between the 12 mitochondrial protein-coding gene sequences. Preliminary analysis of a nuclear protein-coding gene (pepck) did not reveal any peculiar changes between both groups which suggests that these variants may only differ in their mitochondrial makeup.

Molecular study of Cysticercus tenuicollis from slaughtered sheep in Sulaymaniyah province, Iraq

IntroductionCysticercosis 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 MethodsDNA 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.ResultsFive 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.ConclusionsFour new strains of T. hydatigena were discovered for the first time in the study area.

Morphometric, genetic diversity and phylogenetic analysis of Taenia hydatigena (Pallas, 1766) larval stage in Iranian livestock

Cysticercus tenuicollis as metacestode of Taenia hydatigena is the most prevalent taeniid species in livestock. Eighty-eight C. tenuicollis samples were collected from sheep (n = 44) and goats (n = 44) of the northern Iran from 2015 to 2016. The isolated parasites were characterized by morphometric keys. The DNA of the larval stage was extracted, amplified and sequenced targeting mitochondrial 12S rRNA and Cox 1 markers. A significant difference in larval rostellar hook length was observed in 12S rRNA haplotypes. Analysis of molecular variance of 12S rRNA indicated a moderate genetic diversity in the C. tenuicollis isolates. The pairwise sequence distance of C. tenuicollis showed an intra-species diversity of 0.3–0.5% and identity of 99.5–100%. Using the 12S rRNA sequence data we found a moderate genetic difference (Fst; 0.05421) in C. tenucollis isolates collected from livestock of the northern and southeastern regions of Iran. We concluded that the genetic variants of C. tenuicollis are being undoubtedly distributing mostly in different parts of Iran. Further studies with a larger number of T. hydatigena isolates collected from various intermediate and definitive hosts are needed to study this evolutionary assumption and also to determine the apparent genetic differences observed in the studied regions.