Gastrointestinal parasites of dogs and foxes in the Zanjan province of Iran: With an emphasis on Echinococcus species.

Canids harbor many zoonotic parasites and play an important role in the spread of parasites in the human environment. Estimation of parasitic infection among canids as definitive hosts may help competent authorities design efficient control programs. This study was conducted to determine the prevalence of intestinal parasites in dogs and foxes with an emphasis on Echinococcus spp. A total of 500 fecal samples of dogs and 30 fecal samples of foxes were studied in the summer, autumn, and winter of 2021 in the Zanjan province using the formalin-ethyl acetate concentration technique, followed by multiplex PCR. The overall prevalence of gastrointestinal parasite infection was estimated to be 19.05%. The prevalence was 24.8%, 10.2%, and 26.7% in stray, shelter dogs and foxes, respectively. No parasites were found among pet and guard dog samples. PCR results on Taenidae eggs showed that 2.4% of samples were positive for Echinococcus granulosus and none contained E. multilocularis. Noteworthy is that E. granulosus was identified only in stray dog samples. The higher prevalence of E. granulosus infection in stray dogs in this province emphasizes the importance of monitoring the food sources consumed by these animals.

DEVELOPMENT OF A TRIPLEX REAL-TIME PCR ASSAY TO DETECT ECHINOCOCCUS SPECIES IN CANID FECAL SAMPLES.

Echinococcosis is a zoonotic disease with great significance to public health, and appropriate detection and control strategies should be adopted to mitigate its impact. Most cases of echinococcosis are believed to be transmitted by the consumption of food and/or water contaminated with canid stool containing Echinococcus spp. eggs. Studies assessing Echinococcus multilocularis, Echinococcus granulosus sensu stricto, and Echinococcus shiquicus coinfection from contaminated water-derived, soil-derived, and food-borne samples are scarce, which may be due to the lack of optimized laboratory detection methods. The present study aimed to develop and evaluate a novel triplex TaqMan-minor groove binder probe for real-time polymerase chain reaction (rtPCR) to simultaneously detect the 3 Echinococcus spp. mentioned above from canid fecal samples in the Qinghai-Tibetan Plateau area (QTPA). The efficiency and linearity of each signal channel in the triplex rtPCR assay were within acceptable limits for the range of concentrations tested. Furthermore, the method was shown to have good repeatability (standard deviation ≤0.32 cycle threshold), and the limit of detection was estimated to be 10 copies plasmid/µl reaction. In summary, the evaluation of the present method shows that the newly developed triplex rtPCR assay is a highly specific, precise, consistent, and stable method that could be used in epidemiological investigations of echinococcosis.

Geographical Detector-based influence factors analysis for Echinococcosis prevalence in Tibet, China.

Echinococcosis, caused by genus Echinococcus, is the most pathogenic zoonotic parasitic disease in the world. In Tibet of the People's Republic of China, echinococcosis refers principally to two types of severe zoonosis, cystic echinococcosis (CE) and alveolar echinococcosis (AE), which place a serious burden on public health and economy in the local community. However, research on the spatial epidemiology of echinococcosis remains inadequate in Tibet, China. Based on the recorded human echinococcosis data, maps of the spatial distribution of human CE and AE prevalence in Tibet were produced at city level and county level respectively, which show that the prevalence of echinococcosis in northern and western Tibet was much higher than that in other regions. We employ a geographical detector to explore the influencing factors for causing CE and AE while sorting information on the maps of disease prevalence and environment factors (e.g. terrain, population, and yak population). The results of our analysis showed that biological factors have the most impact on the prevalence of echinococcosis, of which the yak population contributes the most for CE, while the dog population contributes the most for AE. In addition, the interaction between various factors, as we found out, might further explain the disease prevalence, which indicated that the echinococcosis prevalence is not simply affected by one single factor, but by multiple factors that are correlated with each other complicatedly. Our results will provide an important reference for the evaluation of the echinococcosis risk, control projects, and prevention programs in Tibet.

Economic Importance of Hydatid Cyst in Slaughtered Cattle of Mugla Province

Objective: This study was carried out to investigate the prevalence, seasonal distribution and economic effects of hydatid cyst infection in slaughtered cattle of Mugla province. Methods: Data from the 2019 records of the Mugla Metropolitan Municipal Abattoir were studied retrospectively. Both direct and indirect calculation methods were used to determine the economic losses incurred due to hydatid cyst. Results: Twenty-one out of 9.985 (0.21%) cattle were found to have been infected with hydatid cyst in 2019. The highest prevelance of infection was reported in February (1.17%), while no cases were observed in May, June and August. The direct economic loss attributed to liver destruction in Mugla province was 11,760 TL (1.950 $). The total indirect economic loss caused by hydatid cyst (carcass loss, milk production loss, decreased fecundity) was 122,691 TL (20,346 $). Cumulatively, the direct and indirect economic losses for hydatid cyst disease in Mugla province were 134,451 TL (22,296 $). Conclusion: According to our results, the prevalence of hydatid cyst in cattle of Mugla province was 0.21%, which was still lower than the average loss in Turkey in spite of the corresponding significant economic loss.

Concomitant Presence of Hydatid Cyst and Colorectal Liver Metastasis

A 65-year-old man, with signs of acute colon obstruction, was diagnosed with rectal tumour and liver hydatid cyst. Additionally, a focal liver lesion in segment 1 was detected. Moreover, physical examination revealed hepatomegaly and abdominal distension. Thus, rectal resection and small liver lesion biopsy was performed. Serological and pathohistological analyses showed concomitant presence of hydatid cyst and colorectal metastasis in the liver. Hence, the cyst was treated with anthelmintic therapy, and patient lived another year after the diagnosis. To the best of our knowledge, cases of concomitant hydatid cyst and colorectal liver metastasis has never been reported; thus, this article addresses a unique case of coexistence between these two serious liver diseases.

The situation of echinococcosis in stray dogs in Turkey: the first finding of Echinococcus multilocularis and Echinococcus ortleppi.

Echinococcosis, caused by larval stage of the genus Echinococcus, is one of the most important zoonotic diseases worldwide. The purpose of this study was to determine the presence and prevalence of Echinococcus species in stray dogs of Erzurum, a highly endemic region for cystic echinococcosis (CE) and alveolar echinococcosis (AE) in Turkey. The study samples consisted of 446 stray dog faecal specimens collected from an animal shelter in Erzurum, Turkey, between October 2015 and February 2016. The faecal samples were collected from individual dogs for the isolation of taeniid eggs using the sequential sieving and flotation method (SSFM). Molecular analyses and sequencing revealed the prevalence of Echinococcus spp. as 14.13% (63/446) in faecal samples. The stray dogs harboured five different Echinococcus spp.: E. granulosus s.s. (G1/G3) (n = 41), E. equinus (G4) (n = 3), E. ortleppi (G5) (n = 1), E. canadensis (G6/G7) (n = 3) and E. multilocularis (n = 16). E. granulosus s.s. was the most abundant species. Surprisingly, the occurrence of E. multilocularis in dogs was revealed for the first time in Turkey. E. ortleppi was also reported for the first time in Turkey. These findings highlight a significant public health risk for human AE and CE, presenting useful baseline data on Echinococcus spp. infection in dogs for designing control strategies.

A 27-Year-Old Man With Intermittent Chest Pain During the Last 4 Years.

CASE PRESENTATION: A 27-year-old man from Eritrea presented to the ED complaining about a progressively worse blunt chest pain in the anterior right hemithorax. Chest pain started 4 years ago and was intermittent. During the last 6 months, symptoms got worse, and the patient experienced shortness of breath in mild exercise. For this purpose, he visited another institution, where a chest radiograph was performed (Fig 1). He was advised to visit a pulmonologist for further evaluation, with the diagnosis of a loculated pleural effusion in the right upper hemithorax.

Serology for the diagnosis of human hepatic cystic echinococcosis and its relation with cyst staging: A systematic review of the literature with meta-analysis.

BACKGROUND: The diagnosis of cystic echinococcosis (CE) is primarily based on imaging, while serology should be applied when imaging is inconclusive. CE cyst stage has been reported among the most important factors influencing the outcome of serodiagnosis. We performed a systematic review and meta-analysis of the relation between cyst stage of hepatic CE and diagnostic sensitivity of serological tests, to evaluate whether their relation is a consistent finding and provide guidance for the interpretation of results of serological tests. METHODOLOGY/PRINCIPAL FINDINGS: MEDLINE, EMBASE, CENTRAL, and Lilacs databases were searched on December 1st 2019. Original studies published after 2003 (year of publication of the CE cyst classification), reporting sensitivity of serological tests applied to the diagnosis of human hepatic CE, as diagnosed and staged by imaging, were included. The quality of studies was assessed using the Newcastle-Ottawa Scale. Data from 14 studies were included in the meta-analysis. Summary estimates of sensitivities and 95% confidence intervals were obtained using random effects meta-analysis. Overall, test sensitivity was highest in the presence of CE2 and CE3 (CE3a and/or CE3b), and lowest in the presence of CE5 and CE4 cysts. ELISA, ICT and WB showed the highest sensitivities, while IHA performed worst. CONCLUSIONS/SIGNIFICANCE: The results of our study confirm the presence of a clear and consistent relation between cyst stage and serological tests results. Limitations of evidence included the heterogeneity of the antigenic preparations used, which prevented to determine whether the relation between cyst stage and sensitivity was influenced by the type of antigenic preparation, the paucity of studies testing the same panel of sera with different assays, and the lack of studies assessing the performance of the same assay in both field and hospital-based settings. Our results indicate the absolute need to consider cyst staging when evaluating serological results of patients with hepatic CE.

Meta-analysis of the prevalence of Echinococcus in dogs in China from 2010 to 2019.

BACKGROUND: Echinococcosis (canine Echinococcus disease) is a neglected tropical disease that causes serious public harm. Dogs, as a terminal host of Echinococcus spp., are a key part of the Echinococcus epidemic. Echinococcosis spreads easily in humans and animals in some areas of China and it is therefore necessary to fully understand the prevalence of Echinococcus spp. in dogs. METHODOLOGY/PRINCIPAL FINDINGS: PubMed, ScienceDirect, Chongqing VIP, China National Knowledge Infrastructure (CNKI), and WanFang databases were searched for relevant articles published in the past 10 years. A final total of 108 studies were included. The overall prevalence of Echinococcus spp. in dogs in China was 7.3%, with the highest point estimate found in sampling year 2015 (8.2%) and publication year 2015 (16.5%). Northwestern China (7.9%) had the highest infection rate in China. Qinghai Province (13.5%) showed the highest prevalence among the 11 provinces we included. We also found that geographical and climatic factors are related to the incidence of canine echinococcosis. We further investigated the source of heterogeneity by analysis of subgroups (sampling district, detection method, dog type, season, parasite species, medication, and study quality level). CONCLUSIONS/SIGNIFICANCE: Our research indicated that Echinococcus spp. were still prevalent in some areas in China. More localized prevention and control policies should be formulated, including improving drinking water hygiene and strengthening hygiene promotion. We recommend the rational use of anti-Echinococcus drugs. In addition, treatment of livestock offal and feces and improving the welfare of stray dogs may play an important role in reducing canine Echinococcus infections.

Echinococcus shiquicus in Qinghai-Tibet plateau: population structure and confirmation of additional endemic areas.

Echinococcus shiquicus is currently limited to the Qinghai­Tibet plateau, a large mountainous region in China. Although the zoonotic potential remains unknown, progress is being made on the distribution and intermediate host range. In this study, we report E. shiquicus within Gansu and Qinghai provinces in regions located not only around the central areas but also the southeast edge of the plateau and describe their genetic relationship with previous isolates from the plateau. From 1879 plateau pikas examined, 2.39% (95% CI 1.79­3.18) were infected with E. shiquicus. The highest prevalence of 10.26% (4.06­23.58) was recorded in Makehe town, Qinghai province. Overall the prevalence was marginally higher in Qinghai (2.5%, CI 1.82­3.43) than in Gansu (2%, CI 1.02­3.89). The cox1 and nad1 genes demonstrated high and low haplotype and nucleotide diversities, respectively. The median-joining network constructed by the cox1­nad1 gene sequences demonstrated a star-like configuration with a median vector (unsampled haplotype) occupying the centre of the network. No peculiar distinction or common haplotype was observed in isolates originating from the different provinces. The presence of E. shiquicus in regions of the southeast and northeast edges of the Qinghai­Tibet plateau and high genetic variation warrants more investigation into the haplotype distribution and genetic polymorphism by exploring more informative DNA regions of the mitochondrial genome to provide epidemiologically useful insight into the population structure of E. shiquicus across the plateau and its axis.

Cystic echinococcosis in sheep and goats of Lebanon.

Cystic echinococcosis (CE), a zoonotic disease caused by the species complex of Echinococcus granulosus sensu lato (s.l.), is endemic in Middle East and the Mediterranean basin, where pastoral activity is widespread. Despite the chronic endemicity of the disease in Lebanon and neighbouring countries, recent data are scant. The objectives of this survey were to evaluate the current epidemiology of CE in Lebanon, investigate the prevalence and determine the infecting genotypes in locally raised sheep and goats. A multidimensional approach combining post-mortem inspection of slaughtered animals and molecular diagnosis of the parasite was conducted to this end. From 2018 to 2020, 62.9% of the sheep and 20.9% of the goats were found positive for CE. The presence of hydatids varied between organs, showing higher prevalence in the liver of sheep vs the lungs of goats, however, a higher fertility rate of hydatid cyst was observed in lungs for both animals. Molecular diagnosis based on partial mitochondrial cox1 gene (795 bp) showed that the majority of isolates were identified as E. granulosus sensu stricto (98.7%) and only one isolate from goat was identified as Echinococcus canadensis (genotype G7; 1.3%). Echinococcus granulosus s.s. population among the sheep and goats was represented by 22 haplotypes having very little genetic differentiation and relatively moderate haplotype diversity. Population demographics explored through neutrality indices suggested expanding population within the intermediate hosts. These results document the high prevalence of CE in the livestock of Lebanon and reveal for the first time the presence of three different genotypes G1, G3 and G7.

First description of the emergence of Echinococcus ortleppi (G5 genotype) in sheep and goats in Iran.

Cystic echinococcosis is one of the most important cyclozoonotic helminthic diseases caused by various genotypes of Echinococcus granulosus SENSU lato complex in the Middle East. Echinococcus ortleppi G5 genotype has been previously reported to infect camels in Iran. However, no molecular data are available on the circulation of the cattle genotype from other animal intermediate hosts. Overall, 30 hydatid cyst samples were collected from 2015 to 2016 from sheep (n; 15) and goat (n; 15) isolates in Mazandaran province, northern Iran. The DNA of larval stages was extracted, amplified and sequenced by targeting of mitochondrial 12S rRNA marker. Based on sequencing and phylogenetic analyses, 28 isolates were identified as E. granulosus G1. However, two fertile hydatid cyst samples isolated from sheep and goat liver were confirmed as E. ortleppi with 99.8%-100% identity to species isolated from a buffalo in India. The first emergence of fertile cattle cysts from sheep and goats may have implications in the zoonotic importance of E. ortleppi in human infection and de-worming regime of infected dogs due to the shorter maturation time of G5 genotype compared with G1 genotype. Since the distribution of E. ortleppi in Iran is not fully known, further studies are urgently needed to appraise the evolutionary and epidemiology scenarios. These studies can ascertain the transmission dynamics of E. ortleppi inferred by full-length concatenated mitogenomes on the domestic and wildlife hosts, as well as humans in wider areas of Iran and the neighboring countries.

Prevalence of Liver Hydatidosis and Its Economic Significance in Sheep Slaughtered in a Private Abattoir in Konya

Objective: This study aimed to determine the prevalence of liver hydatidosis in sheep slaughtered in a private slaughterhouse in Konya and to estimate the economic loss incurred because of the disease. Methods: The study was conducted over a period of 12 months between 1 June 2018 and 31 May 2019. Given that the aim of this investigation was to determine the prevalence of liver hydatidosis, only the livers of 41,002 sheep were examined for hydatid cysts. Results: The liver of 810 (1.97%) sheep was found to be infected with hydatid cysts during the study period. The infection rate was determined as 5.34% in animals older than one year of age and 1.68% in animals less than one year of age. Regardless of the age group, the highest infection rate was found in autumn (3.34%), while the lowest infection rate was seen in spring (0.84%). In the sheep, the highest infection rate was in December (17.2%), and in lambs, it was in June (2.9%). On the other hand, the lowest infection rate in sheep was observed in November (1.8%), while the lowest infection rate in lambs was found in April (0.7%). The total economic loss incurred due to the annihilated livers was estimated as 36,450 TL (6.417$). Regardless of the number of cysts and degree of infection, the infected livers were completely discarded. The economic loss incurred due to the discarded livers was estimated by considering the 2019 offal prices. Conclusion: Based on the data obtained from this study, it could be concluded that hydatidosis still exists in Konya as well as throughout Turkey and that it causes serious economic loss.

Comparative Evaluation of the Efficacy of Indirect Hemagglutination Test and Radiological Methods in the Pre-diagnosis of Cystic Echinococcosis

Objective: Cystic echinococcosis (CE) is a common public health concern in Turkey. In this study, we investigated the agreement between the results of radiological imaging methods and serological tests through a 5-year retrospective evaluation in patients admitted to a university hospital with a suspicion of CE so as to determine the frequency of CE in the study region. Methods: The indirect hemagglutination test (IHA) results of 1.046 patients obtained from various clinics with the suspicion of CE between January 2014 and December 2018 were retrospectively analysed. Of these, patients with at least one radiological imaging report in the system (938 patients) were included in the study. Radiological imaging findings and IHA test results were compared and examined. Results: Seropositivity was detected by IHA test in 143 (15.2%) of 938 patients included in the study. The CE findings were recorded in at least one radiological imaging report in 130 (90.9%) of 143 patients with positive IHA test. At least one of the radiological imaging reports suggested presence of CE in 362 (38.5%) of all the patients. Conclusion: Thus, serological test and radiological imaging methods should be used in combination for the diagnosis of CE.

Giant Brain Hydatid Cyst in an Adult: A New Case Report

Cystic echinococcosis is an important zoonotic disease that occurs in humans and mammals in general, which causes considerable economic loss and poses health concerns in different parts of the world. The patient involved in this case report was a 28-year old man living in Birjand city who had been suffering from intermittent headache, nausea and vomiting for the past two weeks. The other symptoms presented by the patient were dizziness, blurred vision, seizures and imbalance. The patient only complained of headache in the last two weeks and had no symptoms of visual or speech impairment. He had a history of consuming raw vegetables, but did not have canine contact. In brain computed tomography and magnetic resonance imaging, a large cyst was evident in his brain. The patient was admitted to Razi Hospital in Birjand and followed-up by surgical treatment of the hydatid cyst, with no complications observed.

Copro-polymerase chain reaction has higher sensitivity compared to centrifugal fecal flotation in the diagnosis of taeniid cestodes, especially Echinococcus spp, in canids.

Prompt and reliable diagnostic tests for taeniid infection in canids are important due to the risk of zoonoses like Echinococcus spp. Current diagnostic methods relying on fecal flotation lack sensitivity and specificity, but this has rarely been quantified due to the challenges in performing adult cestode recovery (the gold standard) in domestic dogs (Canis familiaris). Therefore, we recovered adult Taenia and Echinococcus spp. from intestines, as well as fecal/intestinal material from 484 wild canids trapped for fur in two Canadian provinces (276 foxes - primarily Vulpes vulpes, coyotes - Canis latrans, and wolves - Canis lupus in Québec and 208 coyotes in Saskatchewan). The performances of a newly developed coproPCR for tapeworm DNA detection in dogs, and centrifugal fecal flotation using Sheather's solution, were evaluated against adult cestode recovery. Overall, adult taeniid cestode prevalence (Taenia and/or Echinococcus) was 28 % (95 % CI: 23-33 %) in Québec (62 % (CI: 51-73%) of 74 coyotes, 65 % (CI: 44-82) of 23 wolves, and 11 % (CI: 7-16%) of 179 foxes) and 79 % (CI: 73-84%) of 208 coyotes in Saskatchewan. In Québec, E. canadensis and Taenia spp. were detected in coyotes and wolves, and foxes were only infected with Taenia spp., whereas Saskatchewan coyotes were predominantly infected with E. multilocularis (at significantly higher prevalence, but not intensity, than coyotes in Québec). Compared with centrifugal fecal flotation, the new coproPCR had at least double the sensitivity (58 % vs 23 % in QC coyotes, 57 % vs 23 % in QC wolves, 24 % vs 0% in QC foxes, and 80 % vs 25 % in SK coyotes). Notably, no taeniid eggs were detected on flotations from foxes infected with Taenia spp., and the new coproPCR had highest sensitivity in Saskatchewan coyotes, which were predominantly infected with E. multilocularis. CoproPCR has promising prospects for use in Veterinary clinics and diagnostic laboratories to detect taeniid cestode infections because of its higher sensitivity than faecal flotation methods. This is particularly important for zoonotic Echinococcus spp. where, from a public health perspective, false negatives are a much greater concern than false positives.

Human and animal cystic echinococcosis in Konya, Turkey: molecular identification and the first report of E. equinus from human host in Turkey.

Cystic echinococcosis is a neglected, zoonotic disease in Turkey. The disease is commonly seen in rural areas where the local population is in close contact with livestock and dogs. This research aimed to molecularly identify of hydatid cysts in cattle and human isolates from Konya, Turkey. Following sample collection, direct microscopy was performed. After direct examination, total DNA was extracted, and positive PCR products of cox 1 mitochondrial gene (~ 875 bp) were sequenced. A total of 83 hydatid cysts (cattle n = 57 and human n = 26), 82 were identified as Echinococcus granulosus sensu stricto (G1-G3 genotypes), and one human isolate was characterized as Echinococcus equinus (G4 genotype). Fertility rates of cysts belonging to cattle for liver and lung cysts were 93.3% and 80%, respectively. Out of 26 human originated isolates, 18 (69.2%) of cysts were found to be fertile. To the best of our knowledge, this is the first report of E. equinus from human host in Turkey.

Liver graft harbouring hydatid disease: how far can we extend our donor pool?

As the current demand for liver transplantation exceeds our donor pool, the donor search is shifted towards the extended donor criteria. The livers harbouring hydatid disease are a controversial source of grafts. We report the use of a liver graft harbouring hydatid disease in urgent liver transplantation in a patient with autoimmune hepatitis. Corroborated with previous experiences, we show that the liver grafts harbouring hydatid cysts provide a rare but valuable source of organs.

Evaluation of 170 Followed-up Cases Treated for Hydatid Disease: A Multicentre Study.

OBJECTIVE: To determine the clinical characteristics of patients with cystic echinococcosis and the diagnostic and therapeutic approaches used. METHODS: This is a multicentre, retrospective study. Patients from six centres who were diagnosed with hydatid cysts in the last five years were evaluated. RESULTS: The mean age was 45.4±17.4 years, and 54.7% were female. The most common complaints were abdominal pain, nausea and vomiting, and the most common physical examination finding was abdominal tenderness. Most patients were diagnosed within 2-6 months. Anaemia and eosinophilia were the most common laboratory findings. The liver was the most commonly involved organ (n=153, 90%). One hundred twenty-five (73.5%) patients underwent ultrasonography. The largest cyst was present in the liver at stage four, and its diameter was 160x170 mm. The rates of the negative, grey zone and positive results were 9.4%, 8.8% and 81.8%. Surgery was more common (n=72, 42.4%) than puncture, aspiration, injection, and re-aspiration treatments (n=14, 8.2%). Of the 47 patients who had a recurrence, 22 (46.8%) had a history of hydatid cyst treatment. CONCLUSION: Hydatid disease, which is endemic in our country, is diagnosed at a late stage. In terms of disease diagnosis, serological tests may be misleading. It was not possible to compare the success rates according to the treatment choices because of the lack of randomisation of stages and sizes of the lesions.