Control Of Echinococcosis Research Articles

Quantifying the load of Echinococcus granulosus eggs in experimental dog infection using probe-based copro-qPCR analysis.

Designing and implementing Cystic Echinococcosis control programs require quantitative information about the worm load and the intensity of infection in dog populations in endemic areas. So far no "probe-based" molecular quantification tool has been available for E. granulosus. This study was conducted in order to develop and evaluate a qPCR technique for measuring worm load of E. granulosus in the final host. A species-specific TaqMan probe was designed based on the available sequences in GenBank. The study was conducted in two stages. First, stool samples from an experimentally infected dog were collected in days 7, 14, 21, 28, 35 and 49, and were analyzed by real-time qPCR assay. In the second stage, 600mg negative stool specimens were manually spiked with 1, 5, 10, 20 and 40 eggs and the specimens were analyzed using real-time qPCR. According to the standard curve analysis, 93% efficiency and coefficients of correlation (Rsq) > 0.991 were documented. Quantitative PCR assays showed an increasing signal of infection during the 7-week course of infection. As revealed by the qPCR results from week 5 onward, signals indicative of egg excretion began and reached maximum on week 7. No qPCR signal from the samples containing 1, 10 and 20 eggs was recorded, however the samples containing 5 and 40 eggs produced signals proportional to the primary DNA. The study presents a molecular tool to quantify the burden of E. granulosus infection in dogs. This tool could be applied for measuring the burden of infection in the definitive hosts in surveillance and control programs.

Intestinal Expression of miR-130b, miR-410b, and miR-98a in Experimental Canine Echinococcosis by Stem-Loop RT-qPCR

Echinococcus granulosus is a zoonotic cestode dwelling in the small intestine of canid definitive hosts. Intermediate hosts are a wide range of domestic and wild ungulates. Human infection with the larval stage causes cystic echinococcosis. Understanding the nature and extent of molecular mechanisms involved in host–parasite interactions helps to answer some very basic questions in the biology of cestode parasites with significant implications in the management and control of cystic echinococcosis. Little is known on the miRNAs expression in the intestinal tissues of dogs infected with E. granulosus. In the present study, expression of a selected profile of miRNAs was evaluated in experimental canine echinococcosis. MiRNAs were extracted from 20 different parts of small intestinal tract of two sibling 3-months-old mix-breed dogs. Complementary DNA was specifically synthesized using an optimized stem-loop system. Intestinal expression of four miRNAs (cfa-let7g, cfa-miR-98, cfamiR-410, cfa-miR-130b) was evaluated using RT-qPCR. The results of the study indicate a significant difference between test and control dogs in cfamiR-130b, cfa-miR-98, and cfa-miR-410 (P ≤ 0.05); however, there was no significant difference for cfa-let7g. The most upregulated miRNAs were cfamiR-130b and cfa-miR-98. An increasing trend for cfa-let7g and a declining trend for cfa-miR-98, cfa-miR-410, and cfamiR-130b were found toward the distal segments of the small intestine. Our study revealed that cfa-miR-98, cfa-miR-410, and cfamiR-130b are involved in the definitive host response in canine echinococcosis. The study provides new information on the molecular basis of interactions between E. granulosus and dogs in terms of miRNA expression and showed that E. granulosus infection could increase the expression of some pro-inflammatory miRNAs at the cellular level in the definitive host.

Identification of vulnerable populations and knowledge, attitude, and practice analysis of echinococcosis in Tibet Autonomous Region of China

Echinococcosis is one of the 17 neglected tropical diseases recognized by the World Health Organization and causes a huge global disease burden. The Tibet Autonomous Region (Tibet) of China is the most epidemic area of echinococcosis worldwide. Echinococcosis poses an enormous threat to local public health and economic development. Identifying vulnerable populations and analyzing people's knowledge, attitudes, and practices (KAP) with respect to echinococcosis is necessary to prevent and control echinococcosis. Based on reported cases and questionnaire responses, we first used a k-prototypes clustering algorithm to identify vulnerable populations according to human demographics (including gender, age, education level, and occupation). We then used a KAP analysis to evaluate the Tibetan people's understanding of echinococcosis. The results identified three types of vulnerable population. Vulnerable population I comprised illiterate middle-aged and older women engaged in agriculture and animal husbandry. Vulnerable population II comprised illiterate middle-aged male herdsmen. Vulnerable population III comprised young male farmers with a low education level. The KAP analysis indicated that the pass rate for all participants was 72.6%, 6.4%, 95.0%, and 75.8% for KAP, K, A, and P, respectively. The pass rates were significantly different among different age and gender groups. Based on these results, it is necessary to improve public health education and professional training, and to implement effective control and management measures targeting typical hosts of echinococcosis. This study provides a direct reference for the prevention and control of echinococcosis in Tibet.

In vitro effects of glutathione on Transforming Growth Factor beta and Epidermal Growth Factor Receptor genes expression in the protoscoleces and strobilated worms of Echinococcus granulosus

Cystic Echinococcosis (CE) caused by the small taeniid cestode Echinococcus granulosus, is a globally distributed zoonosis. Administration of some chemicals or natural compounds could lead to significant effects on the expression of some developmentally important genes including Transforming Growth Factor beta (TGF-β) and Epidermal Growth Factor Receptor (EGFR) in other parasitic organisms. The main purpose of this study was to describe the effect of glutathione (GSH) on the expression of TGF-β and EGFR genes in different developmental stages of E. granulosus.Protoscoleces of hydatid cysts collected from naturally infected sheep liver were cultured in diphasic CMRL1066 medium. Glutathione Mono-ethyl Ester (GME) at 250 μg/ml concentration was applied on the invaginated protoscoleces (PSCi), evaginated protoscoleces (PSCe) and strobilated worms (SW3) in vitro. TGF-β and EGFR genes expression were evaluated by using Real Time qPCR analysis compared to the controls.In response to GME treatment TGF-β expression was affected, however no significant effect was observed in EGFR expression. The results indicate a significant difference of TGF-β expression in the intact protoscoleces and the strobilated worms comparing to the controls. In intact invaginated protoscoleces TGF-β expression was significantly increased (p < 0.01) while in the strobilated worms a significant decrease was observed comparing to no-treatment controls (p < 0.001). None of the three developmental stages of E. granulosus demonstrated significant changes in EGFR expression.The results indicated that administration of GSH modified TGF-β expression in the protoscoleces and strobilar stages of E. granulosus. To improve our understanding of the physiology and biochemistry of the parasite more in depth in vitro and in vivo studies on the morphological and molecular effects of glutathione on the parasite is recommended. Further investigation on the gene profiles in other stages including microcysts and germinal layer cells is also suggested. This paves the way for the effective treatment and control of cystic echinococcosis.

SOME RESULTS OF RESEARCH WORK ON PREVENTION OF ECHINOCOCCOSIS IN KYRGYZSTAN

The Kyrgyz Research Institute of Veterinary Medicine is a co-executor of the Strategy for the Echinococcosis Control (2013–2018) and monitored dogs epizootological examinations after preventive treatment with azinox. Laboratory studies of biomaterials (feces) for infection with helminths were carried out along with the traditional coprological method, a more sensitive enzyme-linked immunosorbent assay (ELISA). The results of laboratory studies of fecal samples from dogs showed that the activities of the "Strategy for the Control of Echinococcosis in the Kyrgyz Republic", including the treatment of dogs with azinox, gave positive outcomes. From 2014 to 2018 2260 feces samples from deworming dogs from almost all regions of Kyrgyzstan were examined. At the same time, the infection of dogs with taeniidae is steadily decreasing; it has also been noted that the main joints of parasitocenosis in dogs are taeniidae, toxocaras, mesocestoids, and eimeria. In the studies conducted on the infection of dogs with taeniidae, the coprological and more sensitive ELISA method was used. Using both helminthological studies, a decrease in the invasion of dogs by taeniidae was confirmed. Thus, in the period from 2014 (the beginning of the Strategy) to 2018, the average invasion of dogs was reduced from 20.2 to 14.3%.

Control of Cystic Echinococcosis in Iran: Where Do We Stand?

Cystic echinococcosis (CE) is an important public health challenge in Iran that requires a One Health approach to control. Filling information gaps and research needs on different aspects of the disease, including parasite biology, disease ecology, human/animal infection, and administrative issues, is essential to address challenges faced by CE control programs.

Knowledge, attitude and practice of echinococcosis prevention and control, and their influencing factors among residents living in Seda County, Sichuan Province

To understand the status of knowledge, attitude and practice on echinococcosis prevention and control and investigate their influencing factors among residents living in Seda County of Sichuan Province, so as to provide a scientific basis for the further implementation of health education interventions in the county. A multi-stage stratified random sampling method was employed on September 2018. Ten townships were randomly sampled, and 2 to 3 administrative villages were randomly sampled from each township. Then, 20 to 40 households were randomly sampled from each village, and 1 to 2 villagers were investigated in each household. The knowledge, attitude and practice on echinococcosis prevention and control were investigated using a household questionnaire survey, and the factors affecting the qualification of echinococcosis prevention and control knowledge were identified using chi-square test and logistic regression analysis. A total of 760 questionnaires were assigned, and 748 effective questionnaires were recovered, with a recovery rate of 98.42%. The overall qualification rate of echinococcosis prevention and control knowledge was 56.42% (422/748) in residents living in Seda County, and the awareness of the question "How to get echinococcosis?" was only 48.40%. The proportion of subjects with a positive attitude towards echinococcosis prevention and control was 64.71% to 94.79%, and the rate of corrected echinococcosis prevention and control behaviors was 10.40% to 82.45%. Univariate analysis showed that factors affecting the qualification rate of echinococcosis prevention and control knowledge included gender, occupation, education level, frequency of echinococcosis screening and frequency of health education (all P < 0.05), and binary logistic regression analysis showed that women, herdsmen as the primary occupation, an education level of primary school and below, absence of echinococcosis screening and absence of health education were factors responsible for the unqualified awareness of echinococcosis prevention and control knowledge (all P < 0.05). There is a low awareness rate of echinococcosis prevention and control knowledge and a low proportion of corrected echinococcosis prevention and control behaviors in residents living in Seda County, Sichuan Province. Multiple forms of health education activities pertaining to echinococcosis prevention and control are needed with adaptation to local circumstance, and implementation of appropriate behavioral interventions is recommended.

An old drug as a promising new cure for the hard-to-treat echinococcosis

An old drug as a promising new cure for the hard-to-treat echinococcosis

The effects of dog management on Echinococcus spp. prevalence in villages on the eastern Tibetan Plateau, China

BackgroundThe pastoral area of the eastern Tibetan Plateau is highly endemic for human echinococcosis. Domestic dogs are the main definitive host for the transmission of both Echinococcus granulosus (sensu lato) and E. multilocularis to humans. To control the infection risks, a national-level canine echinococcosis prevention and control programme has been implemented since 2015 in Shiqu County, Ganze Tibetan Autonomous Prefecture, Sichuan, China. The objective of this study was to evaluate its effect on Echinococcus spp. prevalence in dogs.MethodsWe surveyed 69 households with 84 owned dogs, for dog fecal samples and dog keeping information in the villages of Rizha and Eduoma. A total of 105 dog fecal samples (75 from owned dogs and 30 unknown dog fecal samples) were collected between 2015–2017 to determine Echinococcus spp. prevalence using copro-PCR. Eight variables based on household surveys were included into a logistic regression model for significant risk factors to canine echinococcosis prevalence in dogs.ResultsBetween 2015–2017, the overall Echinococcus spp. copro-DNA prevalence decreased significantly in dogs from 51.2% (2015) to 20.0% (2017) in Rizha, and insignificantly from 11.5% (2016) to 4.3% (2017) in Eduoma. Echinococcus multilocularis was the most prevalent species continually copro-DNA detected during the entire study period, while E. granulosus was rare and not detected in 2017. Echinococcus shiquicus copro-DNA prevalence (a probable non-zoonotic wildlife species) was as high in dogs as that of E. multilocularis, although only detected in 2015 in Rizha. Unleashed dog feces were mainly collected in Rizha in 2015. Although 93.2% of owned dogs were leashed, and the monthly praziquantel dosing rate reached 97%, E. multilocularis infection could still be detected in 11.1% of owned dogs in 2017. Monthly deworming, leashing dogs 24 h per day, and the avoidance of dogs feeding on livestock viscera were significant measures to prevent canine echinococcosis infection in owned dogs.ConclusionsCarrying out a canine echinococcosis prevention and control programme can significantly decrease Echinococcus spp. prevalence. The potential contact between leashed dogs and wild small mammals is still a risk for re-infection of owned dogs with E. multilocularis. This study shows that the long-term application of regular dog treatment with praziquantel in the vast and remote echinococcosis endemic areas of the eastern Tibetan Plateau can reduce transmission in dogs but remains a challenging intervention.

Knowledge, attitudes and practices regarding echinococcosis in Xizang Autonomous Region, China

BackgroundEchinococcosis is a neglected zoonotic parasitic disease caused echinococcus parasitizes, poseing a significant economic burden on both humans and animals. There are limited studies on echinococcosis in China, especially Xizang Autonomous Region, although the area is endemic area for echinococcosis. The study aimed to provide information for strategic prevention against this disease.MethodsA cross-sectional survey was conducted among the residents in Xizang Autonomous Region, China to evaluate their knowledge, attitudes and practices on endemicity of echinococcosis. A face-to-face survey was conducted to collect the data using a well-designed questionnaire. The contents included basic personal information, knowledge, attitudes and practices about echinococcosis, personal hygiene and behavior habits, dog feeding and whether they had received the information on echinococcosis, and so on. We surveyed 840 persons in practice. All data analysis was performed using Epi Info 7.2.ResultsOf the total particpants, 86.8% had a primary education level or below (including primary and illiterate), and even 45.0% were illiterate. Farmers and herdsmen represent the main occupations in this study. People who know all the echinococcosis-related knowledge in the questionnaire only accounted for 8.7% of the participants. However, none of the participants was aware of routes of echinococcosis infection in human or dogs. The data showed participants with higher educational background had the high awareness rate of echinococcosis-related knowledge or attitudes (chi-square for trend, χ2 = 21.23, p<0.05 & χ2 = 48.43, p<0.05). In addition, The percentage of the participant with awareness of echinococcosis-related practices was associated with their age and principle occupation (χ2 = 52.72, p<0.05 & χ2 = 20.63, p<0.05).ConclusionsXizang Autonomous Region is an epidemic area of Echinococcosis. The prevalence of the disease has been largely due to the lack of knowledge, awareness, and poor hygiene practice in local residences. Therefore, effective disease prevention education and awareness campaigns in community will be significantly helpful in prevention and control of echinococcosis.

Assessment of the direct economic losses associated with hydatid disease (Echinococcus granulosus sensu stricto) in beef cattle slaughtered at an Australian abattoir

Bovine hydatid disease, characterised by fluid-filled hydatid cysts, is regularly found in the offal of beef cattle at slaughter. Organs found to be infected at slaughter are removed to preclude them from entering the human food chain. The organs are either downgraded to pet food or condemned. Previous studies have focussed on total economic losses, but have not calculated the cost of disease per animal, which would be useful information for producers when determining how best to manage hydatid disease. This study estimated the direct losses associated with hydatid disease in beef cattle slaughtered at an Australian beef abattoir both at the population (all cattle slaughtered) and individual animal level.Data on annual prevalence of hydatid disease in beef cattle were obtained from an Australian abattoir for the years 2011–2017. The direct losses resulting from the condemnation and downgrading of offal infected with hydatid cysts at the abattoir were estimated using data stratified by age, sex and feed-type. Official and literature-based sources of organ weight and price were used to estimate direct losses associated with hydatid disease in beef cattle slaughtered at the abattoir. Uncertainty and variability in input parameters were represented using uniform distributions and Monte Carlo sampling was used to model output parameter uncertainty.Out of 1,097,958 beef cattle slaughtered between January 2011 and December 2017, 97,832 (8.9%) were reported infected with hydatid disease. The median estimated direct loss to the abattoir for the duration of the study period was AU$655,560 (95% confidence interval [CI] AU$544,366–787,235). This equated to approximately AU$6.70 (95% CI AU$5.56–8.05) lost per infected animal. The annual median estimated direct losses due to hydatid disease at the abattoir were AU$93,651 (95% CI AU$77,767–112,462). Direct losses varied each year of the study and ranged from AU$38,683 in 2016 to AU$163,006 in 2014.This estimate of the direct losses associated with bovine hydatid disease most likely underestimates the true extent of the overall losses because indirect losses such as reduced carcass weights were not estimated in this study. Nevertheless, these estimates illustrate the negative economic impact of bovine hydatid disease and demonstrate that improved surveillance to enable control of hydatid disease should be considered both in Australia and globally. It would be worthwhile to estimate the losses in other beef abattoirs for the same time period to compare results, and to investigate the cost-benefit of control programs for bovine hydatid disease.

Contribution to the echinococcosis control programme in China by NIPD-CTDR.

As a zoonotic parasitosis caused by the parasitism of Echinococcus larvae, echinococcosis imposes serious disease and economic burdens on human beings and society, and is thus a global public health issue. Its complex life history, wide distribution, the combined influence of various epidemic factors, coupled with the unique natural environment, customs, and religious beliefs in endemic areas, pose a huge challenge to the national echinococcosis control programme in China. Accurate early detection and confirmation of diagnosis of echinococcosis, the use of effective drugs, real-time surveillance of the infection status of populations and various hosts, controlling the source of infection, and blocking the route of transmission are of enormous significance for control. In this paper, the work by NIPD-CTDR on the prevention and control of echinococcosis in China is reviewed, with a view to providing reference for the further promotion of the national echinococcosis control programme.

Development of a copro-LAMP assay for detection of several species of Echinococcus granulosus sensu lato complex

Cystic echinococcosis represents a significant problem in human and animal health and constitutes one of the most severe Neglected Tropical Diseases prioritized by the World Health Organization. The etiological agent is the complex Echinococcus granulosus sensu lato (s. l.), composed of several species/genotypes. Diagnosis in the definitive host and molecular epidemiology studies are important points for cystic echinococcosis control. Here we developed a new copro-LAMP assay, LAMP EGSL, for diagnosis in the definitive host for simultaneous detection of Echinococcus granulosus sensu stricto (s. s.), Echinococcus ortleppi, and Echinococcus canadensis species. Also, the analytical sensitivity, specificity and plausibility of performance in a rural context of a previously reported species-specific LAMP reaction, was evaluated. Both reactions showed high analytical sensitivity values (10 fg-100 fg DNA) and did not show cross reaction with DNA from host or other helminthic parasites. LAMP EGSL was performed with samples from an endemic area. In addition, the alkaline hydrolysis of one E. granulosus s. s. adult parasite followed by specific LAMP to E. granulosus s. s. was performed in a laboratory with low resources from another cystic echinococcosis endemic area. The results obtained suggest that LAMP EGSL represents a potential tool for canine diagnosis that could be useful for cystic echinococcosis control programs. In addition, we showed that LAMP reaction for E. granulous s. s., E. ortleppi and E. canadensis specific detection, could be useful for molecular epidemiology studies applicable to the definitive host. Both reactions were performed in endemic, rural areas without sophisticated equipment.

First inter-laboratory comparison of Echinococcus granulosus sensu lato diagnosis in Latin America.

ABSTRACTObjective.To compare the performance of polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) tests for diagnosing Echinococcus granulosus in dog feces among national reference laboratories in Argentina, Chile, Peru, and Uruguay.Methods.National laboratories affiliated with the Ministry of Health/Agriculture of each country exchanged panels of 10 positive/negative samples obtained from their regular national surveillance programs in November 2015 – November 2016. All laboratories applied PCR; two also applied ELISA techniques. Sensitivity and specificity were determined for each laboratory and concordance of results among the laboratories was evaluated by Cohen Kappa coefficient.Results.Poor concordance (3 of 10 paired comparisons had values of Kappa > 0.4), low sensitivity and specificity across all laboratories, and poor performance of both techniques in detecting E. granulosus in canine feces was demonstrated in this study. An ex-post comparison of the laboratories’ test protocols showed substantial heterogeneity that could partially explain poor concordance of results.Conclusion.The results underscore the heterogeneity of canine echinococcosis diagnosis across the region and indicate possible sources of variability. Efforts to standardize canine echinococcosis testing must be included in the plan of action for the Regional Initiative for the Control of Cystic Echinococcosis. Future comparisons with fecal samples of known parasite load are needed.

Risk factors of human cystic echinococcosis in Shiqu County Sichuan Province: a case-control study

To identify the main risk factors of human cystic echinococcosis in Shiqu County, Sichuan Province, so as to provide the reference for the formulation of echinococcosis control strategies in Shiqu County. During the period from November 2015 through June 2017, the patients with cystic echinococcosis (case group) and healthy controls (control group) were randomly sampled from Shiqu County as the study subjects. A questionnaire survey was conducted to capture the study subjects'age, gender, ethnicity, occupation, religion, lifestyle, education level, number of household dogs, bovine and sheep, and density of dog feces in the courtyard. The major risk factors of human cystic echinococcosis were identified using a logistic regression model. Univariate logistic regression analysis showed 7 risk factors with statistical significance between the case and control groups, and age, lifestyle, number of household bovine, number of household sheep, number of house-hold dogs, and the density of dog feces in the courtyard were included in the multivariate logistic regression model (OR = 1.026, 4.792, 1.067, 1.022, 1.709 and 1.095, respectively). High age, pastoral nomadic lifestyle, high number of house-hold bovine, high number of household sheep, high number of household dogs and high density of dog feces in the courtyard are strongly associated with the riks of human cystic echinococcosis in Shiqu County.

Prevalence of bovine hydatidosis and its economic importance in Adama Municipal Abattoir, Eastern Ethiopia

Hydatidosis is a major parasitic disease condition responsible for low productivity of livestock industry in Ethiopia due to imposing poor weight gains and condemnation of organs. This study was conducted from November 2016 to April 2017 in Adama municipal abattoir to estimate the prevalence, organ distribution of the cyst, and economic importance of bovine hydatidosis. Routine meat inspection, hydatid cyst count and characterization were conducted. Out of 400 cattle slaughtered at Adama municipal abattoir, 210 (52.5%) animals were found harboring hydatid cysts. The infection prevalence was higher in lung 105(26.2%) than the rest of the organs examined. In the present study, explanatory variables (sex, age, breed, body condition and origin) were found to have statistically significant (p&lt;0.05) association with the prevalence of hydatidosis. Of the total of 970 hydatid cysts counted, 529/731(72.4%) small, 89/100(89%) medium, 42/45(93.3%) large and 20/94(21.3%) calcified cysts were detected in lung. Liver harbored 194/731(26.5%) small, 11/100(11%) medium, 3/45(6.7%) large and 74/94(78.7%) calcified cysts. Out of 208 cysts subjected for fertility and viability testing 120(57.7%), 74(35.6%), 40(54%) 34(46%) and 14(6.7%) were sterile, fertile, viable, non-viable and calcified cysts, respectively. The financial loss due to organ condemnation annually and during the study period was estimated to be 894,505.612 ETB and 17,844.90 ETB, respectively. The prevalence of hydatidosis in such a high proportion implies that the disease is economically important in the study area. Therefore, there should be strict control of hydatidosis by focusing on backyard slaughter practice, unsafe offal feeding of dogs, and proper west disposal. Moreover, awareness creation on its zoonotic importance, route of transmission and the growing stray dog population need to be given special attention.Keywords: Abattoir; Adama; Cattle; E. granulosus; hydatidosis

Spatial and temporal distribution of newly diagnosed echinococcosis patients in Sichuan Province from 2007 to 2017

To explore the spatial and temporal distribution characteristics of cases with newly diagnosed echinococcosis in Sichuan Province from 2007 to 2017, so as to provide reference for the formulation of echinococcosis prevention and control strategies and for the identification of key areas. The spatial distribution maps of detection of cases with newly diagnosed echinococcosis were plotted in Sichuan Province from 2007 to 2017, and the spatial distribution characteristics and epidemic trends were analyzed. From 2007 to 2017, the detection rate of cases with newly diagnosed echinococcosis appeared a decline in Sichuan Province year by year, and the areas with a high detection rate of cases with newly diagnosed echinococcosis were mainly located in western, northwestern and northern parts of Sichuan Province, while the areas with a low detection rate were predominantly found in the southern and eastern parts of the province. The global Moran's I values were 0.19, 0.22, 0.17, 0.44, 0.48, 0.31 and 0.16 for the detection rate of cases with newly diagnosed echinococcosis in Sichuan Province from 2010 to 2016 (all Z scores > 1.96, all P values < 0.05), suggesting spatial aggregation distribution during this period. Local spatial autocorrelation analysis revealed that the"high-high"areas and"low-low"areas for the detection rate of cases with newly diagnosed echinococcosis all showed an aggregation tendency. The detection rate of cases with newly diagnosed echinococcosis decreases in Sichuan Province from 2007 to 2017 year by year, and shows a spatial aggregation. The echinococcosis control activities should be intensified in Shiqu, Seda, Dege, Ganzi and Baiyu counties.

Control of cystic echinococcosis: Background and prospects.

Cystic echinococcosis (CE), caused by Echinococcus granulosus, is a chronic and debilitating zoonotic larval cestode infection in humans, which is principally transmitted between dogs and domestic livestock, particularly sheep. Human CE occurs in almost all pastoral communities and rangeland areas of the underdeveloped and developed world. Control programmes against CE have been implemented in several endemic countries to reduce or eliminate the disease. New Zealand and Tasmania are examples of some of the first programmes to be undertaken (in insular territories) and which were very successful in the elimination of CE. The advent and proven effectiveness of praziquantel, plus the experience of insular models, produced high expectations for rapid advances in a second generation of control programmes undertaken in continental areas (Argentina, Uruguay and Chile). Nevertheless, only moderate gains in CE control have been made and the impact on prevalence among humans has been slight. A major impediment to the adoption of procedures that were successful in New Zealand and Tasmania has been the requirement to administer praziquantel to dogs in rural areas eight times per year over numerous years. In addition, there have been clear technological improvements made in the diagnosis of canine echinococcosis for surveillance, the genetic characterization of parasite strains and in vaccination against CE infection in livestock. In order to establish new paradigms and appropriate combinations of control strategies, we have carried out a review and discussion of the available control tools and control models. Control strategies must be suitable and sustainable to benefit the Echinococcosis-endemic areas primarily, which at the same time are the poorest regions of the world.

Isolated Human and Livestock Echinococcus granulosus Genotypes Using Real-Time PCR of cox1 Gene in Northeast Iran.

Cystic echinococcosis (CE), caused by the metacestode of Echinococcus granulosus, is highly endemic over large parts of Iran. The purpose of this study was to investigate the prevalence rate of hydatidosis and mitochondrial cox1 real-time PCR with high-resolution melting curve (HRM) analysis of E. granulosus isolated from human and livestock. In this cross-sectional study, 61 formalin-fixed paraffin-embedded tissue isolates were collected from human CE cases and 83 hydatid cysts from the liver and lung lesions of the livestock in Khorasan Razavi province, Northeast Iran. DNA was extracted from each isolate and amplified by real-time PCR and analyzed using the HRM method. The HRM analysis using the cox1 gene of 40 E. granulosus human isolates showed that 35 (87.5%), 4 (10%), and 1 (2.5%) of the isolates were categorized as G1, G3, and G6 genotypes, respectively. Out of the total 1342 livestock inspected, 39 (4%) goats and 44(12%) cattle were found harboring hydatid cysts all belonging to E. granulosus sensu stricto. The results confirmed that the high prevalence of E. granulosus sensu stricto in intermediate hosts is remarkable in northeast of Iran coupled with the high prevalence of infection in livestock, which reinforced the need for hydatidosis control programs in this region.

A multiplex PCR for differential detection of Echinococcus granulosus sensu stricto, Echinococcus multilocularis and Echinococcus canadensis in China

BackgroundEchinococcosis caused by Echinococcus is one of the most major infectious diseases in north-west highland of China. E. granulosus sensu strict, E. multilocularis, and E. canadensis are known to be the only three species related to human health transmitting in the areas. To achieve targeted treatment and control of echinococcosis, the accurate identification and discrimination of the species are important. However, currently the available diagnostic approaches do not present ideal results either in accuracy or efficiency.MethodsIn the study, a set of primers were designed to aim at the three human-pathogenic Echinococcus species in China. The one-step multiplex PCR assay was developed and evaluated for the specificity and sensitivity. A total of 73 parasitic lesions and 41 fecal materials obtained from human and various animals collected in the clinic and the field were tested to assess the applicability of this method.ResultsThe multiplex PCR effectively detected the individual DNA from the targeted species and their random mixtures generating with distinguishable expected size of products. The detection limit of the assay for each of the three species was 5 pg/μl when they were tested separately. When DNA mixtures of the targeted species containing the same concentration were used as templates, the lowest amount of DNA which can be detected was 50 pg/μl, 10 pg/μl and 5 pg/μl for E. granulosus s. s., E. multilocularis, and E. canadensis respectively. No cross-reactivity was observed when DNA from eight genetically close species was used as control templates. The multiplex PCR identifications of all samples were in line with the original sequencing results except for those infected with E. shiquicus, which showed negative signals in the developed assay. Of all the tested stool materials, 16 were previously found positive for Echinococcus by visual and microscopic examination. Among these 16 samples, 13 were confirmed by the multiplex PCR, and the other three tested negative. Additionally, the multiplex PCR identified another 14 positive feces from the remained 25 stool samples which absence of worms.ConclusionsThe developed multiplex PCR shows advantages in fast diagnosis and large-scale epidemiological investigation, which proven to be a promising tool utilized in clinic and surveillance system.