Validation of deep amplicon sequencing of Dicrocoelium in small ruminants from Northern regions of Pakistan.

Dicrocoelium lancet flukes cause significant production loss in ruminant livestock. Although co-infection with multiple Dicrocoelium species within a host is common, techniques for studying the composition of these complex parasite communities are lacking. The pathogenicity, epidemiology, and therapeutic susceptibility of different helminth species vary, and little is known about the interactions that take place between co-infecting species and their hosts. Here, we describe the first applicationof metabarcoding deep amplicon sequencing method to studythe Dicrocoelium species in sheep and goats. First, rDNA ITS-2 sequences of four Dicrocoelium species (Dicrocoelium dendriticum, Dicrocoelium hospes, Dicrocoelium orientalis, and Dicrocoelium chinensis) were extracted from the NCBI public database. Phylogenetic analysis revealed separate clades of Dicrocoelium species; hence, molecular differentiation between each species is possible in co-infections. Second, 202 flukes belonging to seventeen host populations (morphologically verified as belonging to the Dicrocoelium genus) were evaluated to determine the deep amplicon sequencing read threshold of an individual fluke for each of the four species. The accuracy of the method in proportional quantification of samples collected from single hosts was further assessed. Overall, 198 (98.01%) flukes were confirmed as D. dendriticum and 1.98% produced no reads. The comparison of genetic distances between rDNA ITS-2 revealed 86% to 98% identity between the Dicrocoelium species. Phylogenetic analysis demonstrated a distinct clustering of species, apart from D. orientalis and D. chinensis, which sit very close to each other in a single large clade whereas D. hospes and D. dendriticum are separated into their own clade. In conclusion each sample was identified as D. dendriticum based on the proportion of MiSeq reads and validated the presence of this group of parasites in the Gilgit Baltistan and Khyber Pakhtunkhwa provinces of Pakistan. The metabarcoding deep amplicon sequencing technology and bioinformatics pathway have several potential applications, including species interactions during co-infections, identifying the host and geographical distribution of Dicrocoelium in livestock, drug therapy response evaluation and understanding of the emergence and spread of drug resistance.

Histopathology and antibody responses describe the seasonal pattern of dicrocoeliosis in small ruminants in the Himalayan ranges of Pakistan.

In some parts of the world, Dicrocoelium spp. lancet flukes cause significant production loss in pastoral livestock, and accurate diagnosis of infection is important. The aims of the present study were to describe the histopathology and to investigate the transmission patterns of Dicrocoelium amongst ten sheep and goat farms in Khyber Pakhtunkhwa and Gilgit Baltistan, Pakistan. The liver histology and indirect enzyme-linked immunosorbent assay (ELISA) analyses followed standard procedures. The liver histopathology showed intensive tissue destruction and biliary hyperplasia associated with presence of adult flukes, severe inflammatory cell infiltration, congestion of blood vessels, damaged hepatocytes, and sinusoids in the infected areas. The time of onset of infection was investigated by ELISA detection of antibodies in sheep (n = 164) and goats (n = 152). Colostral transfer of Dicrocoelium antibodies from seropositive mothers was detected in sheep and goats up to 16 weeks of age. In both sheep and goats, the estimated time of infection differed between farms and years. Infection was seen in both sheep flocks and goat herds, with high variation between flocks and herds, and the highest infection rate in lambs. Dicrocoelium infection was most prevalent in sheep and goats in September (n = 84) and August (n = 63) respectively. This study concluded Dicrocoelium causes severe inflammation and necrosis of liver tissues in sheep and goats. Colostral transfer of antibodies can be detected up to about ten weeks of age. Higher infection rates are observed during August and September in sheep than in goats, putatively due to effects of different grazing and browsing behaviors on the ingestion of ants. The results will aid in the development of effective disease control strategies to ensure optimal growth and productivity of sheep and goats.

Zoonotic parasite infection from a funerary context: A Late Antique child case from Cantabrian Spain.

OBJECTIVE: To evaluate the presence of Dicrocoelium sp. in a child from a Late Antique funerary context from Cantabrian Spain and discuss whether the infection is true infection or pseudoparasitosis. MATERIALS: Four skeletons, including one from a 5-7 year old child, have been analysed from the archaeological site of El Conventón, dated between the sixth and seventh centuries AD. METHODS: The paleoparasitological study was conducted through the analysis of soil samples from different parts of the skeleton and funerary context using the rehydration, homogenization, and micro-sieving method, and visualized through brightfield microscopy. RESULTS: A soil sample from the pelvic region tested positive for Dicrocoelium sp. (possibly D. dendriticum). CONCLUSIONS: The child was infected with Dicrocoelium dendriticum, which based on archaeological and historical contexts may be related to hygiene or dietary behaviour. SIGNIFICANCE: We present one of the few cases of the identification of a Dicrocoelidae parasite directly associated with a human skeleton that provides historical knowledge of a zoonotic disease. LIMITATIONS: The diagnosis of a zoonosis through the identification of ancient parasites is complex. In addition, Dicrocoelium sp. in association with skeletal human remains is rare due to the potential low prevalence of this parasite. SUGGESTIONS FOR FURTHER RESEARCH: Highlight the importance of paleoparasitological analysis to link parasitic infection diseases with socioeconomic issues by using funerary contexts with skeletal remains.

Spatial Distribution of Dicrocoelium in the Himalayan Ranges: Potential Impacts of Ecological Niches and Climatic Variables.

PURPOSE: Dicrocoeliosis can be an important cause of production loss in ruminants due to the cost of liver condemnation at slaughter. The aim of the present study was to determine the prevalence of Dicrocoelium infection and to predict the ecological niches and climatic variables that support dicrocoeliosis in the Himalayan ranges of Pakistan. METHODS AND RESULTS: Dicrocoelium was detected in 33 of 381 liver samples and 238 of 6060 blood samples taken from sheep and goat herds in the area. The prevalence of dicrocoeliosis was higher in sheep than in goats and highest in females aged more than 3 years. An environmental risk map was created to predict active zones of transmission and showed the highest probability values in central parts of the Chitral district in the northwest of Pakistan. Climatic variables of the mean monthly diurnal temperature range (Bio2), annual precipitation (Bio12), and normalised difference vegetation index (NDVI) were found to be significantly (p < 0.05) associated with the presence of Dicrocoelium infection. CONCLUSION: Together, the findings of this study demonstrate the most suitable ecological niches and climatic variables influencing the risk of dicrocoeliosis in the Himalayan ranges of Pakistan. The methods and results could be used as a reference to inform the control of dicrocoeliosis in the region.

A survey of sheep dicrocoeliosis in Sulaymaniyah slaughterhouse, northern Iraq in 2013-2014.

Dicrocoelium dendriticum is a helminth which lives in the bile ducts and gall bladder of mammalian definitive hosts. Dicrocoeliosis is thought to be endemic in some countries and it has an increasing prevalence in Northern Iraq, potentially due to increased importation of infected animals, particularly sheep, in high numbers from neighboring countries. The parasite's ability to infect rodents, wild animals, livestock and humans means this parasite is of significant interest in veterinary and human medicine. While D. dendriticum causes relatively mild clinical disease in animals, infection leads to liver condemnation at slaughter and subsequent economic losses to farmers. In this study, the livers of 91,486 sheep slaughtered at Sulaymaniyah New Slaughterhouse (Northern Iraq) were visually inspected for D. dendriticum infection between November 2013 and March 2014, with 1269 livers rejected due to D. dendriticum in this period, representing 29.4% of all condemned livers. The highest rate of rejection due to D. dendriticum infection was seen in December, possibly linked to increased numbers of intermediate host snails during the wet season. Routine inspection of condemned livers revealed adult flukes of D. dendriticum and eggs were seen in the faeces of imported sheep, indicating these animals may be a vector for introduction of D. dendriticum to this region. Due to the complex life cycle and wide range of animals which it infects, we cannot establish the exact route of introduction into Iraq; however, our study suggests that slaughterhouse workers, farmers and local health authorities should be aware of the presence of D. dendriticum, and the potential risks it represents to both human and animal health. Our data also suggest that some level of quarantine or border checks may be useful to prevent further introduction of D. dendriticum or other pathogens into Iraq, although this may prove difficult until accurate diagnostic assays are developed.

Evaluation of zoonotic platyhelminthe infections identified in slaughtered livestock in Iran, 2015-2019.

BACKGROUND: Platyhelminth infections of livestock can result in considerable economic losses. This study aims to evaluate the spatial frequency of cystic echinococcosis (CE), dicrocoeliasis, and fascioliasis in livestock slaughtered in Iran during the years 2015-2019 and estimate direct costs associated with organ condemnation due to these parasites. METHODS: Abattoir data from 413 abattoirs representing all 31 Iranian provinces were collected from the Iran Veterinary Organization. Infection prevalence was calculated per year at the province level. The Local Moran's I statistic was performed to evaluate spatial autocorrelation of animals positive at slaughter for the years 2015-2019. Direct costs associated with condemned livers were calculated for each parasitic condition, with costs associated with condemned lungs also included for CE. RESULTS: Overall prevalence values for the study timeframe were as follows: sheep and goat fascioliasis 1.56% (95% CI: 1.56-1.56%), cattle fascioliasis 3.86% (95% CI: 3.85-3.88%), sheep and goat dicrocoeliasis 4.63% (95% CI: 4.62-4.63%), cattle dicrocoeliasis 3.08% (95% CI: 3.07-3.09%), sheep and goat CE 5.32% (95% CI: 5.32-5.33%), and cattle CE 7.26% (95% CI: 7.24-7.28%). Northwest Iran had the highest prevalence of CE and fascioliasis. High infection areas for Dicrocoelium spp. included the provinces of Zanjan, Gilan, Qazvin, and Tehran, which are located in northern Iran. Direct economic losses for sheep and goat fascioliasis, dicrocoeliasis, and CE for the study period were US$13,842,759, US$41,771,377, and US$22,801,054, respectively. Direct economic losses for cattle fascioliasis, dicrocoeliasis, and CE for the study period were US$1,989,200, US$1,668,986, and US$2,656,568, respectively. CONCLUSION: Our findings provide valuable data for future monitoring of these important parasitic diseases in Iranian livestock. Disease control strategies are required to reduce the economic and public health impact of these platyhelminths.

Parasitic causes of meat and organs in cattle at four slaughterhouses in Sistan-Baluchestan Province, Southeastern Iran between 2008 and 2016.

This 8-year (from 2008 to 2016) retrospective study calculated the percentage of carcass and organ (lung and liver) condemnations and estimated the direct financial costs at four slaughterhouses in Sistan-Baluchestan Province, Southeastern Iran. Each carcass and organ (lung and liver) was thoroughly examined through inspection, palpation and incision following the standard protocol. Identification of the parasites was performed macroscopically. The total direct economic loss due to meat's condemnation was estimated by adding weights of each organ or carcass part and multiplying individual organ totals by their 2016 market unit price. A total of 857,039 cattle were slaughtered during this period, 64,497 livers (7.5%), 31,401 lungs (3.6%) and the carcasses of 1,171 cattle (0.1%) were condemned due to lesions caused by parasites. The main parasitic lesions in the condemned livers were attributed to Echinococcus granulosus (4.2%), Fasciola spp. (3.1%) and Dicrocoelium dendriticum (0.1%). All the condemned lungs were due to E. granulosus (3.6%). Taenia saginata cysticerci were detected in 0.1% of inspected animals. Liver condemnation due to cystic echinococcosis was the highest in fall (4.7%, p < 0.001); while lung condemnation was the highest during spring (3.98%, p < 0.001). Liver condemnation due to Fasciola spp. was the lowest in winter (2.99%, p < 0.001). Carcass condemnation as a result of cysticercosis was the highest in summer (p < 0.001). Considering the 2016 market prices, condemnations due to the studied parasites caused direct costs estimated U.S. $ 3,191,879. To the best of our knowledge, this is the first report estimating the monetary losses due to parasitic infections in the slaughterhouses of this province. Due to the high financial impact of the studied parasites, a control programme should be implemented to decrease this impact.

Dicrocoelium spp. in cattle from Wa, Ghana: prevalence and phylogeny based on 28S rRNA.

Dicrocoeliosis is a trematode infection in cattle, sheep and goats caused by the small liver fluke, Dicrocoelium spp. Though endemic in Ghana, its disease situation is poorly understood. In the present study, the prevalence, distribution and worm load of Dicrocoelium spp. in cattle at slaughter in Wa were determined. A total of 389 cattle were screened during meat inspection for liver flukes, and polymerase chain reaction accompanied by DNA sequencing of the 28S rRNA gene was used to identify Dicrocoelium spp. Generally, prevalence of bovine dicrocoeliosis (small liver fluke) stood at 19.54 % with prevalence in males and females being 17.62 % and 21.43 %, respectively. Animals under 2 years suffered more infection than older ones (23.08 % vs. 16.80 %). Dicrocoelium infection was recorded in animals from all the communities where slaughtered cattle came from. On average, 31 flukes per infected animal were recorded. A molecular confirmatory test on seven flukes identified them as D. hospes. This preliminary study highlights the importance of bovine dicrocoeliosis in Ghana and has identified D. hospes as a causal agent. The data provides basis for further studies to appraise the trematode disease situation in animals and phylogeny of Dicrocoelium spp. circulating in Ghana.

Prevalence of Liver Trematode Infections in Cattle in the Province of Agri in Turkey

Objective: This study was conducted to determine the prevalence of distomatosis in cattle in the Agri province in Turkey. Methods: The livers and bile ducts of 200 slaughtered cattle were examined macroscopically, and the gall bladders were examined by sedimentation. The presence of Fasciola hepatica eggs and antigens in 188 cattle were investigated using sedimentation and coproantigen ELISA, respectively. Egg counts per gram of faeces (EPG) was determined using the modified McMaster sedimentation method. Results: Adult Fasciola hepatica and Dicrocoelium dendriticum were found in the bile ducts of 47 and 25 cattle, respectively. Examination of the gall bladders of cattle revealed the presence of eggs of Fasciola hepatica and D. dendriticum in 63 and 48 cattle, respectively. The coproantigens of Fasciola hepatica were found in 148 (78.7%) cattle using ELISA. Additionally, the eggs of F. hepatica were found in 63 (33.5%) cattle by sedimentation, with the value of EPG between 17 and 83. The prevalence of D. dendriticum was determined as 25.5%, with the value of EPG between 17 and 67. Conclusion: In this study, distomatosis was detected in cattle in Agri province using coproantigen ELISA and sedimentation methods along with macroscopic examination. The findings necessitate an urgent implementation of an effective prevention and control program, which can increase the awareness of cattle breeders regarding these diseases. Furthermore, there is also a need for more detailed studies on these diseases.

Prevalence of hydatid cyst, Fasciola spp. and Dicrocoelium dendriticum in cattle and sheep slaughtered in Sabzevar abattoir, Iran.

Hydatid cyst, Dicrocoelium dendriticum, and Fasciola spp. are common parasites among ruminants. The current study was conducted to assess the prevalence of these parasites in cattle and sheep slaughtered in Sabzevar abattoir, from April 2014 to March 2016. Total of 1653 cattle and 26720 sheep were inspected for helminthic infections of liver and lung in different seasons. The prevalence rates were found to be 684 (2.55%) and 235 (14.21%) in sheep and cattle, respectively. The infection rate of liver in cattle was estimated to be 5.86%, 0.84% and 5.68% for hydatid cysts, Fasciola spp. and D. dendriticum, respectively and lung infection with hydatid cyst in cattle was 1.81%. Moreover, the liver infection rate of sheep was estimated to be 0.87%, 0.06% and 2.13% with hydatid cysts, Fasciola spp. and D. dendriticum, respectively and lung infection with hydatid cyst was 0.35%. The rate of infection with D. dendriticum was higher in the both animals than the other parasites. The results of this study indicated that the prevalence of the aforementioned parasites was high, which caused high economic losses. Therefore, sanitary and preventive measures should be taken.

Predicting environmental suitability and geographical distribution of Dicrocoelium dendriticum at littoral of Caspian Sea: An ecological niche-based modeling.

Dicrocoeliasisis caused by the small liver fluke (Dicrocoelium spp.), mainly Dicrocoelium dendriticum in domestic and wild ruminants. The small liver fluke is the probable predisposing cause of economic burden. The impact of geographic and climatic factors on the incidence of dicrocoeliasis has been severely ignored in different geographical areas. Due to the lack of data regarding dicrocoeliasis in Iran, this study was aimed to investigate the prevalence and intensity of ovine and bovine Dicrocoelium infection in the coastal strip south of the Caspian Sea. Fecal samples were obtained from the cattle and sheep in three provinces of Guilan, Mazandaran and Golestan at the littoral of the Caspian Sea. All collected samples were then tested by flotation methods for determining the number of eggs per gram of feces (EPG). Moreover, we applied maximum entropy niche-based modeling (MaxEnt), coupled with remote sensing and the Geographical Information System (GIS) to visualize the spatial distribution and risk factors of Dicrocoelium dendriticum at the littoral of Caspian Sea. A total of 2688 stool samples were collected from cattle (n = 1344) and sheep (n = 1344) in coastal provinces of the Caspian Sea including Guilan (n = 1280), Mazandaran (n = 768) and Golestan (n = 640) provinces. Based on the data presented here, the highest rate of infection was observed in Guilan and Mazandaran provinces. The results revealed the prevalence rates of 36.72% and 6.09% for sheep and cattle in Guilan province, respectively. This rate was 22.4% for sheep and 3.91% for cattle in Mazandaran province. However, the rate of sheep infection was 90% in some point locations. Dicrocoelium infection was found to be significantly different between three provinces in sheep (P < 0.00001, Chi = 111.633). Our findings exhibited a high reliability of the MaxEnt model, and area under the curve (AUC) values of the training and test data sets were determined to be 0.852 and 0.818, respectively. Jackknife analysis showed the relative variable contribution to the model performance, where four variables were found as key influential factors that highly affected the habitat suitability of the presence of the lancet fluke including the precipitation of driest quarter (Bio17), altitude, temperature seasonality (Bio4), and precipitation of driest month (Bio14). The findings of this study demonstrated a high presence rate of Dicrocoelium infection at the littoral of Caspian Sea, Iran. Moreover, climatic variables can be considered as important predictive factors affecting the distribution of infection in the studied areas. Further studies based on the findings of the GIS are also very important in the country for planning control programs.

Dicrocoeliosis in extensive sheep farms: a survey.

BACKGROUND: This study investigated the epidemiological and molecular aspects of dicrocoeliosis in extensive sheep farms. METHODS: From 2013 to 2014, copromicroscopical analyses in 190 dairy sheep farms and anatomo-pathological inspections in six slaughterhouses were carried in Sardinia, Italy. Rectal faecal samples were analyzed using the FLOTAC® method, and anatomo-pathological examinations were based on detecting thickened terminal bile ducts (TTBDs). In addition, genetic analyses were conducted on representative DNA samples of adult Dicrocoelium spp. RESULTS: Ninety-seven (51.1%) out of 190 sheep farms were coprologically positive for Dicrocoelium spp. In the liver, on the surface and cut surface, TTBDs were reported in 40.1% (309/770) and 15.3% (118/770) of the animals examined, respectively, with an overall prevalence of 25.5% (196/770). No intraspecific genetic variation was observed among the Dicrocoelium dendriticum isolates. CONCLUSIONS: Our survey reveals the widespread presence of D. dendriticum in Sardinia, although seasonal, geographical and climatic conditions might be key factors in modulating the infection prevalence. Examining typical lesions due to D. dendriticum in the liver in abattoirs can be used as a marker for tracking chronic dicrocoeliosis infection.

A Comprehensive Retrospective Study: Intestinal Parasites in Human in Van Province

Objective: The aim of present study was to retrospectively determine the prevalence of intestinal parasites in patients, who were referred to Parasitology Laboratory in Van Yüzüncü Yil University, Faculty of Medicine during an 11-year period. Methods: Stool samples of 69633 individuals admitted to the outpatient clinics in the hospital were initially evaluated with native-Lugol, and then by flotation, sedimentation, trichrome staining, modified acid-fast staining and ELISA methods. Results: Twenty-four parasite species were identified in parasite-positive patients. At least one or more parasite species were found in 34.1% of all patients. The most commonly observed parasite was Blastocystis hominis (26.5%). Among pathogen parasites, Giardia intestinalis (G. intestinalis) was detected in 9.3%, Ascaris lumbricoides (A. lumbricoides) was detected in 2.5%, Entamoeba histolytica/Entamoeba dispar in 0.8%, Cystoisospora belli in 0.004%, Fasciola hepatica in 0.04%, Dicrocoelium dendriticum in 0.001%, Strongyloides stercoralis in 0.001% and hookworm in 0.001% of the patients. Conclusion: It was determined that, pathogen parasites such as G. intestinalis and A. lumbricoides were still observed at high rates in Van province, especially in children, and the problem of parasitosis is still continuing, although the prevalence of parasites has declined when compared to the previous years.

Importance of Land Snails in Dicrocoeliosis Epidemiology.

Dicrocoeliosis is a helminthosis caused by the small liver fluke Dicrocoelium spp. (Trematoda, Dicrocoeliidae) parasitizing in the bile ducts and gall bladder of ruminants as well as many other animal species including humans. In the biological life cycle of Dicrocoelium, land snails are first intermediate hosts and ants are second intermediate hosts. Sporocysts and cercaria, which are larval stages, live in the hepatopancreas of land snails and metacercaria, which is also the larval stage, lives in the abdomen and brain of ants. Land snails, which are the first intermediate host of this parasite in Turkey, include Helicopsis derbentina, Helicopsis protea, Helicopsis krynickii, Cernuella virgata, Trochoidea pyramidata, Cochicella acuta, Monacha carthusiana, Helicella candicans, Helix aspersa, Helix lucorum, and Chondrus tournefortianus. Dicrocoeliosis is widespread in ruminants and affects their liver, which can lead to weight loss and reduced milk production. The number of reports on dicrocoeliosis is increasing due to the expansion of dry habitats and parasites becoming resistant to antihelminthic drugs. This study provides information on the epidemiology and control methods of Dicrocoelium.

Population genetic analysis informs the invasion history of the emerging trematode Dicrocoelium dendriticum into Canada.

Parasite distributions are constantly changing due to climate change, local and global movement of animals and humans, as well as land use and habitat change. The trematode Dicrocoelium dendriticum is a relatively recent invader of Canada, being first reported in eastern Canada in the 1930s and western Canada in the 1970s. However, historical records are scarce and its emergence is poorly understood. The establishment of this parasite in Canada provides an interesting opportunity to explore the use of population genetic approaches to help elucidate the invasion history of a relatively recently established helminth parasite. In this study, we compare the genetic diversity and population structure of a number of D. dendriticum populations from western and eastern Canada, and compare these with much longer established European populations. Two independent genetic marker systems were used; a microsatellite marker panel and a cytochrome c oxidase 1 (cox1) mitochondrial (mt)DNA sequence marker. We found distinct differences in both genetic diversity and population structure of the different Canadian populations that provide insights into their invasion histories compared with the European populations. Two populations from British Columbia, Canada - Salt Spring and Vancouver Islands - are of low diversity, show evidence of a population bottleneck and are closely related to each other, suggesting a shared recent history of establishment. These west coast populations are otherwise most closely related to those from eastern Canada and western Europe, and in contrast are genetically divergent from those in Cypress Hills, Alberta, Canada. Although the Alberta parasite population is the most recently reported in Canada, being first identified there in the early 1990s, it was the most genetically diverse of those examined and showed a strong pattern of admixture of genotypes present in western and eastern Europe. Overall, our results are consistent with a model in which western Europe is likely the source of flukes on the east coast of Canada, which were then subsequently translocated to the west coast of Canada. The most recently reported D. dendriticum population in Canada appears to have a different history and likely has multiple origins.

Dicrocoelium dendriticum: An Unusual Parasitological Diagnosis in a Reference International Health Unit.

Finding Dicrocoelium dendriticum eggs in human feces is exceptional and there are few prevalence data available. True infection occurs after accidental ingestion of ants containing metacercariae and spurious infection through the consumption of infected animal liver. Differential diagnosis between true and pseudo-infections is performed through stool examination after a diet free of liver. In addition, microscopy can help to differentiate the type of infection. We report six cases, all from sub-Saharan Africa, detection of this fluke at the Tropical Medicine Unit Vall d'Hebron-Drassanes (Barcelona, Spain). Dicrocoelium dendriticum transit eggs were visualized in five cases and there were no subsequent visualizations after diet, which reinforces that all these cases were false parasitism. In one case, few embryonated eggs were observed and the patient was treated for a possible true parasitism. There is a need to investigate the prevalence of D. dendriticum in our country focusing on the distinction between true and spurious infections.

Characterization of nine microsatellite loci for Dicrocoelium dendriticum, an emerging liver fluke of ungulates in North America, and their use to detect clonemates and random mating.

This study characterizes polymorphic microsatellite loci from adults of the liver fluke Dicrocoelium dendriticum sampled from a population of sympatric beef cattle and wapiti in a region of emergence in southern Alberta, Canada. We also scrutinized the markers to validate their use in studying the population genetics of this complex life cycle parasite. Among the nine loci described, four deviated significantly from Hardy Weinberg Equilibrium (HWE) due to technical artefacts. The remaining five loci were in HWE. These five provided sufficient resolution to identify clonemates produced from the obligate asexual reproduction phase of the life cycle in snails and to assess the impact of non-random transmission of clonemates on measures of FIS, FST and genotypic disequilibrium. Excluding clonemates, we show that the sub-population of worms was in HWE, that average FIS within hosts was 0.003 (p=0.4922) and that there was no population genetic structure among hosts FST=0.001 (p=0.3243). These markers will be useful for studies of Dicrocoelium dendriticum ecology, transmission, and evolution.

PREVALENCE OF DICROCOELIUM DENDRITICUM INFECTION IN SHEEP AT TAIF PROVINCE, WEST SAUDI ARABIA.

Dicrocoelium dendriticum is a common zoonotic parasite of sheep in many Regions of Saudi Arabia. In chronic infections, this parasite causes biliary cirrhosis in livers of cattle, sheep, or goats and leads to economic losses. This study compared the prevalence of Dicrocoelium dendriticum in local sheep and imported sheep in Taif Province, West Saudi Arabia. This cross-sectional survey was carried out in slaughterhouses in Taif Province. A total 2230 local sheep and 21383 imported sheep were studied. The number of injury in imported sheep was observed as 99 representing 0.46% of the number of animals examined, while no record of any injury was observed in local sheep.

Pulmonary Arterial Lesions in New World Camelids in Association With Dicrocoelium dendriticum and Fasciola hepatica Infection.

In Switzerland, dicrocoeliasis is regarded as the most significant parasitic infection of llamas and alpacas. Fasciola hepatica infestation is also a problem but less common. The aim of the present retrospective study was to evaluate the lungs of New World camelids (NWCs) for evidence of arterial hypertension in association with liver changes due to liver fluke infestation. The lungs of 20 llamas and 20 alpacas with liver fluke infestation were histologically evaluated. The hematoxylin and eosin and van Gieson (VG)-elastica stains as well as immunohistology for the expression of α-smooth muscle actin (α-SMA) were used to visualize the structures of arterial walls. Parasitology of fecal matter (11 llamas and 17 alpacas) confirmed that most of these animals were infested with both Dicrocoelium dendriticum and other gastrointestinal parasites. In most cases (10/12 llamas, 4/6 alpacas), liver enzyme activity in serum was elevated. Histologically, arteries in the lungs of 9 of 20 llamas (45%) and 3 of 20 alpacas (15%) showed severe intimal and adventitial and slight to moderate medial thickening, which was confirmed with α-SMA and VG-elastica staining. All animals exhibited typical liver changes, such as fibrosis and biliary hyperplasia, in association with the presence of liver flukes. This study shows that liver flukes can induce proliferative changes in lung arteries in NWCs that resemble those seen with pulmonary arterial hypertension due to liver parasites in humans. However, the degree of liver fluke infestation was not correlated with the extent of liver damage, or with the amount of thoracic or abdominal effusion or pulmonary arterial changes.