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1.
The aim of this study was to estimate the relevance of Echinococcus multilocularis coproantigen detection in fox faeces collected in the field to identify different levels of endemicity for Echinococcus multilocularis on a large scale (n×10 km2). Six study sites were selected in a high endemicity area and two study sites in a low endemicity area in eastern France on the basis of landscape composition. Sampling was undertaken in the winters of 1996–97, 1997–98 and 1998–99. At each site, (i) necropsy and intestine examination was undertaken on a sample of shot foxes (total number of foxes, 222), and (ii) fox faeces were collected in the field along road verges, and scored for degradation status (total number of faeces, 625). Fox faeces were also sampled in a control area (n=30) in western France in the summer of 1998. Intestines were examined according to the sedimentation method. Echinococcus multilocularis coproantigens were detected by using two ELISA tests: EM-ELISA and EmA9-ELISA. The necropsy prevalence in high and low endemicity areas was 63.3% and 19.4%, respectively, and the distribution of adult worms in the fox population was highly overdispersed (75.5% of the total biomass was harboured by 11.6% of foxes). Using the two ELISA tests, there was no difference in the detection of E. multilocularis coproantigens in field faeces, regardless of the degradation status. The medians of EM- and EmA9-ELISA OD values of field faeces in high endemicity area were significantly higher than in low endemicity area (P<0.001 for both ELISA). The distribution of EM-ELISA OD values in low endemicity area was significantly higher (P=0.002) than in the control area. Moreover, for the two ELISA, the observed ELISA OD value distributions in high endemicity area, low endemicity area and control area seemed representative of the distribution of adult worms in fox populations. These results indicate that E. multilocularis coproantigen detection in field faeces could serve for large-scale surveillance, as an alternative to necropsy.  相似文献   

2.
A sandwich ELISA for the detection of Echinococcus granulosus coproantigen in formalin and heat-treated faecal supernatants of dogs was developed. The assay used affinity-purified polyclonal antibodies obtained from rabbits hyperimmunised with E. granulosus excretory/secretory antigens and biotiaylated monoclonal antibody EmA9 produced against adult E. multilocularis somatic extract. The test was sensitive to 7 ng and 2.3 ng of E. granulosus protein and carbohydrate/ml of faecal supernatant, respectively. Thirteen helminth-free dogs were infected with different amounts of E. granulosus protoscoleces and the presence of coproantigen was monitored during the prepatent period until day 35 post-infection, when they were necropsied. Faecal antigen levels started to rise above the normal range between days 10 and 20 post-infection, and typically peaked at the end of the experiment. All the dogs, bearing from 3 to 67 700 worms, showed positive values in the ELISA during the prepatent period. One dog experimentally infected with Taenia hydatigena metacestode and harbouring three worms, tested positive only after the prepatent period at day 52. The test was applied to 98 stray dogs. The ELISA detected all of four dogs naturally infected with E. granulosus, two dogs with patent infections of T. hydatigena and two dogs with no cestode infections, sbowing a sensitivity of 100% and a specificity of 96%.  相似文献   

3.
A sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of Echinococcus multilocularis coproantigens (EM-ELISA) was developed with polyclonal rabbit (solid phase) and chicken egg (catching) antibodies that were directed against E. multilocularis coproantigens and somatic worm antigens, respectively. In experimentally infected dogs and cats, coproantigens were first detectable 6-17 days postinfection (PI) in samples of 8 dogs (worm burdens at necropsy: 6,330-43,200) and from 11 days PI onward in samples of 5 cats infected with 20-6,833 worms. After anthelmintic treatment of 4 dogs and 5 cats at day 20 PI, coproantigen excretion disappeared within 3-5 days. The sensitivity of the ELISA was 83.6% in 55 foxes infected with 4-60,000 E. multilocularis, but reached 93.3% in the 45 foxes harboring more than 20 worms. The EM-ELISA was used in surveys of "normal" dog and cat populations in Switzerland. Among 660 dogs and 263 cats, 5 dogs and 2 cats exhibited a positive reaction. In 2 of these dogs (0.30%) and 1 cat (0.38%), intestinal E. multilocularis infections were confirmed by necropsy, polymerase chain reaction PCR, or both. The specificites of the ELISA in these groups were found to be 99.5% and 99.6%, respectively, if positive ELISA results that could not be confirmed by other methods were classified as "false positive" reactions.  相似文献   

4.
In an epidemiological survey for Echinococcus multilocularis in rodents and insectivores from the northernmost part of the central mainland of Japan (Honshu), two taeniid species, Taenia crassiceps and Taenia polyacantha, were found in Microtus montebelli and Apodemus argenteus, respectively. The latter is the first record of distribution in Japan, and the former is the second after its first recovery from the central part of Japan. Although we have found neither larval nor strobilar stage of E. multilocularis there, discovery of these taeniid species, having overlapping global distribution with E. multilocularis in red foxes, Vulpes vulpes as well as multiple occurrences of hydatid patients having no history of visits to the endemic areas shows the possibility that the life-cycle of E. multilocularis might be maintained at least in the northernmost part of Honshu.  相似文献   

5.
This paper describes the role of fox population level on Echinococcus multilocularis infection in foxes in a highly endemic area in eastern France. Fox population level was monitored by spotlight survey at Le Souillot from 1989 to 2000, and from 1992 to 2000 at Chemin, a control site located in a low endemic area. The infection level of the fox population was estimated at Le Souillot from winter 1995 to winter 1999 using a coproantigen ELISA performed on faeces collected in the field. Population biomass of intermediate hosts (Microtus arvalis and Arvicola terrestris) was monitored using index methods from 1995 to 1999. At Le Souillot, a significant decline in the fox population level was recorded after spring 1997 (P<0.001), and the population level remained low until 2000. The decline occurred when 31% of the grassland area was treated with bromadiolone, an anticoagulant used at a large scale for the control of A. terrestris population outbreaks. No decline of population was recorded at Chemin, where bromadiolone was not used for rodent control. Significant differences among ELISA OD distributions in fox faeces were recorded for the five winters under study at Le Souillot (P=0.0004). The median of ELISA OD distribution was 0.209 and 0.207 before the population decline (winter 1995 and 1996, respectively), significantly increased to 0.306 just after the decline (winter 1997), and then significantly decreased to 0.099 and 0.104 afterwards (winter 1998 and 1999, respectively). Therefore, the decrease in infection level occurred during winter 1998, 1 year after the population decline, when the intermediate host biomass in the field was at its highest. These results suggest a complex dependence between the fox population level and E. multilocularis infection in a high endemicity area. Alternative ways to control fox population as a way to reduce E. multilocularis transmission in a given area are discussed.  相似文献   

6.
Coproparasitological and purging methods for diagnosing canids infected with the intestinal helminth Echinococcus granulosus, an important zoonotic parasite, are unreliable. Detection of coproantigens in feces of infected dogs by enzyme-linked immunosorbent assay (ELISA) is suitable for detecting patent and prepatent infections with a high degree of sensitivity and specificity. In the present study, natural and experimental infections in domestic and wild Australian canids were investigated using a coproantigen capture ELISA. Experimental infection of dogs with E. granulosus was detected at between 14 and 22 days postinfection (PI), and optical density (OD) values remained high until termination of experiments 35 days PI. After chemotherapy, coproantigen levels in infected dogs dropped rapidly, becoming negative 2-4 days after treatment. In experimentally infected red foxes (Vulpes vulpes), the coproantigen excretion profile was different, with ELISA OD levels peaking 15-17 days PI, then falling to low or undetectable levels by 30 days PI. Coproantigens were detected in the feces of naturally infected Australian wild dogs (dingoes, dingo/domestic dog hybrids) with infection levels ranging between 2 worms and 42,600. Preliminary data on the stability of coproantigen in dog feces exposed to environmental conditions indicated that there was no change in antigenicity over 6 days. The results suggest the coproantigen ELISA could be successfully used to monitor E. granulosus prevalence rates in Australian domestic dogs, foxes, and wild dogs.  相似文献   

7.
Survival of protoscolices of Echinococcus multiocularis removed from the intermediate host was studied at constant temperatures. The longest survival time was 16 days at 12°C when the protoscolices were stored within the cyst mass. The infectivity of the protoscolices in the decayed intermediate host was also studied. The carcasses harboring hydatid cysts were fed to dogs and a red fox on the 7th and 14th days after killing. Tapeworms were recovered from all these canines. The results indicate that the protoscolices in the decayed carcasses keep their infectivity for more than 2 weeks and suggest that the saprophagy of canines may be epidemiologically important for the transmission of E. multilocularis.  相似文献   

8.
Coproantigen ELISA based tests for diagnosis of canine echinococcosis provide high specificity and sensitivity. However, the antigenic molecules present in faeces from infected dogs have not yet been characterised. While initial attempts to determine the molecular weights of Echinococcus granulosus coproantigens by SDS-PAGE and Western blotting with coproantigen reactive capture antibodies were equivocal, they suggested presence of a significant carbohydrate component. Periodate treatment of coproantigen positive faecal supernatants resulted in a significant reduction (53%) in ELISA activity, suggesting that carbohydrates are involved in the antigenic structure of E. granulosus coproantigens. Protease treatment of antigenic molecules resulted in an 11% reduction in absorbance in ELISA, indicating that protein components were also present which affected by enzyme activity. Lectin-binding ELISA assays indicated strong affinity of E. granulosus coproantigens to concanavalin agglutinin and Lens culinaris agglutinin, and moderate binding to wheat-germ agglutinin and peanut agglutinin. No binding was detectable to Ulex europaensis agglutinin-I, Bandeiraea simplicifolia or Dolichos biflorus agglutinin. These data indicate that E. granulosus coproantigens from infected dog faeces possibly contained alpha-D-mannose and/or alpha-D-glucose, beta-galactose and N-acetyl-beta-glucosamine residues. To verify the role of carbohydrate moieties in coproantigens, faecal samples were treated with exoglycosidase and tested in the coproantigen ELISA. Treatment with beta-galactosidase or N-acetyl-beta-glucosamine reduced ELISA activity by 44 and 30%, respectively. Incubation with a panel of other specific exoglycosidases including alpha-galactosidase as well as alpha-L-fucosidase, alpha-mannosidase, beta-mannosidase, alpha-glucosidase, beta-glucosidase, beta- fructosidase, or neuraminidase, did not alter coproantigen detection in ELISA. The results indicate that coproantigens present in faeces from E. granulosus naturally infected dogs were highly glycosylated and contain beta- galactose and N-acetyl-beta-glucosamine. The putative relationship of antigenic molecules with the tapeworm glycocalyx is discussed.  相似文献   

9.
Antigen B in hydatid cyst fluid of Echinococcus granulosus is a polymeric lipoprotein of 160 kDa, and is an aggregate of several different but homologous small proteins with approximately 8 kDa which are encoded by a multigene family. Four genes encoding 8-kDa-subunit monomers of the antigen B have been identified from E. granulosus. Recently, we have isolated another novel gene from Echinococcus multilocularis encoding a fifth 8-kDa-subunit of AgB (named EmAgB8/5), predominantly transcribed in the adult worm, but not in vesicles of metacestodes. In this study, we cloned and characterized two EmAgB8/5 homologue genes from E. granulosus genotypes 1 and 6 by PCR, and named as EgG1AgB8/5 and EgG6AgB8/5, respectively. The phylogenetic relationship of these genes with other genes encoding the antigen B 8-kDa-subunit monomers was also discussed.  相似文献   

10.
The sandwich enzyme linked immunosorbent assay (ELISA) was used as a diagnostic test for Echinococcus granulosus infection by detecting coproantigens in 94 stray dogs Canis familiaris and eight red foxes (Vulpes vulpes) from northern Jordan. The results were analyzed in relation to actual helminth infection as revealed by necropsy. The infection rate of dogs with E. granulosus was 13.8% with a worm load ranging between 3-> 10,000 per infected dog. In contrast, eight of 13 E. granulosus infected dogs were coproantigen positive (overall sensitivity 61.5%). The sensitivity increased to 87.5% and 100% in dogs harboring > 20 and > 100 worms/dog, respectively. The specificity of coproantigen-ELISA was 91%. The greatest cross-reactivity was found in dogs infected with Dipylidium caninum. The positive and negative predictive values for the coproantigen-ELISA test were 50% and 94.2%, respectively. Thus, a coproantigen negative dog is most probably truly negative for E. granulosus. In contrast, a coproantigen positive dog may not be truly positive for E. granulosus, except if it has a high worm burden of > 100 worms/animal.  相似文献   

11.
Echinococcus felidis had been described in 1937 from African lions, but was later included in Echinococcus granulosus as a subspecies or a strain. In the absence of any genetic characterization, most previous records of this taxon from a variety of large African mammals remained unconfirmed due to the lack of diagnostic criteria and the possible confusion with the sympatric E. granulosus sensu stricto, Echinococcus ortleppi and Echinococcus canadensis. In this study, we obtained taeniid eggs from lion feces in Uganda and amplified DNA from individual eggs. Mitochondrial and nuclear DNA sequences showed similarities with those of other Echinococcus spp., but high values of percentage divergence of mitochondrial genes indicated the presence of a distinct species. In a second step, we compared this material with the preserved specimens of adult E. granulosus felidis, which had been identified morphologically approximately 40 years ago in South Africa. All DNA fragments (<200 bp) that could be amplified from the adults showed 100% similarity with the Ugandan material. In the phylogenetic tree of Echinococcus which was constructed from the mitochondrial genes, E. felidis is positioned as a sister taxon of E. granulosus sensu stricto. The data obtained will facilitate the development of diagnostic tools necessary to study the epidemiology of this enigmatic parasite.  相似文献   

12.
An epidemiological survey was conducted on the seasonal variation of Echinococcus multilocularis prevalence in red foxes from 1997 to 1998, using a monoclonal antibody-based detection of the tapeworm coproantigen. Thirty-six breeding dens of reproductive fox families were identified in the endemic area of Koshimizu, eastern Hokkaido, Japan. Fecal samples from each site were examined by coproantigen detection assay and fecal egg examination. Whereas the prevalence of coproantigen positive feces showed no seasonal fluctuation (51.6-66.7%), variation was found in the prevalence of egg positive feces in which a higher prevalence was observed in the summer and winter (31.1 and 38.7%) than spring and autumn (13.3 and 13.5%). Significant differences were observed between juveniles and adult foxes in both examinations. Samples from juvenile foxes gave higher coproantigen positive results and taeniid egg intensity. Those results suggest more juveniles infected with the cestode than adults in the same period. The practical use of coproantigen assay as a survey tool and factors which affect the prevalence and host age-related difference are discussed.  相似文献   

13.
To assess the epidemiological efficacy of type SRP III slow-released praziquantel-medicated bars for dogs in the prevention and control of cystic echinococcosis in man and livestock, praziquantel-medicated bars were implanted subcutaneously in over 90% of dogs in villages in north Xinjiang, China, where cystic echinococcosis is highly endemic. After implantation, infection rate of Echinococcus granulosus in dogs, specific antibodies in children and prevalence of echinococcosis in one-year-old lambs were observed for 3 years. Coproantigen of E. granulosus was positive in 41.2% of the dogs at the start of experiment. In the second and third year after medicated-bar implantation, coproantigen was undetectable in any dogs examined, while 3.0% of dogs were positive at the end of the fourth year. The antibody positive rate in 7-year-old pupils, that was 41.2% before the experiment, declined gradually and it was 5.4% in the fourth year, while children in the non-intervention control area showed 30.6% seropositivity. The prevalence of hydatid disease in children aged 7–16 years also declined significantly. The prevalence of hydatidosis in lambs one year of age was 44.8% in the first year, dropping to 10.7% in the fourth year, while in the non-intervention control area the level of infection was 46.4%. These results demonstrated not only that the slow released praziquantel-medicated bars efficiently blocked reinfection in dogs at least for 2 years, but also the measure was effective in preventing transmission of cystic echinococcosis to both man and livestock.  相似文献   

14.
The purpose of this study was to compare the sensitivity and specificity of an ELISA test to detect Fasciola gigantica antigens (coproantigens) in bovine feces, with fecal egg counting and an ELISA for detecting anti-F. gigantica antibodies in serum. Monoclonal antibodies to cathepsin L were generated and used to capture this antigen in feces of infected cattle. Blood, feces, and livers were collected from 150 cattle at an abattoir in Jakarta, Indonesia, for anti-Fasciola antibodies, coproantigen detection, and F. gigantica egg and worm counts. Fluke recovery varied from 1 to 426 per host, with a mean of 32 flukes. The results showed that the sensitivity and specificity of coproantigen detecting ELISA (95 and 91%, respectively) was better than the anti-F. gigantica antibody ELISA (91 and 88%, respectively) and to fecal egg counting (87 and 100%, respectively). The coproantigen ELISA was able to detect 100% of the cattle with >15 flukes. A survey of 305 cattle in central Java over a 10-mo period validated this test in the field, demonstrating a high prevalence of fascioliasis and establishing the test as a useful diagnostic method to determine patent F. gigantica infections in cattle.  相似文献   

15.
The role of the red fox Vulpes vulpes in the dissemination of eggs of Toxocara canis into the environment is considered with reference to female worm fecundity and egg output in the faeces of infected foxes collected from four localities in southern England. A significant positive correlation was found between female worm size and the number of eggs in the uterus but there was no significant relationship between T. canis worm numbers and egg output in fox faeces. Reliable estimates of worm burdens in foxes could not, therefore, be determined from faecal egg counts alone. The highest mean egg output of 2145.0 epg recorded from adult foxes indicated that fox cubs are not necessarily the main sources of environmental contamination with T. canis eggs. Saturated magnesium sulphate was found to be a more effective flotation solution than zinc sulphate and sodium chloride for recovering eggs from fox faecal samples.  相似文献   

16.
In order to identify the respective importance of the ecological and biological factors involved in the transmission of Echinococcus multilocularis, we estimated grassland vole intermediate host (Microtus sp. and Arvicola terrestris) population densities, in relation to the diet of the definitive host (red fox, Vulpes vulpes) and with the prevalence of E. multilocularis in the fox population. The study was conducted in the Ardennes, north-eastern France, which is an area with a high incidence of alveolar echinococcosis. Surface index methods showed that Microtus was the most abundant intermediate host in the area. Furthermore, Microtus was present in one-third of the 144 faeces and 98 stomach content samples examined and represented more than two-thirds of the rodent occurrences. Red fox predation on Microtus was significantly correlated with Microtus relative abundance. In contrast, the relative abundance of A. terrestris was very low. This species, as well as Clethrionomys glareolus and Apodemus sp., was little consumed. E. multilocularis prevalence in foxes was determined from carcasses and reached 53% (95% confidence interval 45-61%). Intensity of infection varied from 2 to 73,380 worms per fox, with 72% of the sampled worm burden harboured by 8% of the sampled foxes. The selected explanatory variables (sex, year, age class, health and nutritional condition, and season) failed to predict prevalence rate and worm burden. The high prevalence rate in foxes indicates the possibility of intense E. multilocularis transmission, apart from periods, or in landscapes, favourable to large population outbreaks of grassland rodents.  相似文献   

17.
, and 1988. Morphological variation of Echinococcus granulosus protoscoleces from hydatid cysts of human and various domestic animals in Jordan. International Journal for Parasitology 18: 1111–1114. Rostellar hook morphology of protoscoleces was employed to study the possible existence of Echinococcus granulosus strains in humans and various domestic animals in Jordan. A distinct form in the donkey was evident as the protoscoleces from this host did not share any characteristics with those from the other hosts examined. Sheep, goats and cattle appeared to be affected by another form since the protoscoleces from their hydatid cysts shared six out of nine characteristics studied. Protoscoleces of camel and human cysts shared seven out of nine characteristics studied and they were different in six characteristics from protoscoleces from other hosts. Differences observed among the three forms may reflect strain variation of E. granulosus in this country.  相似文献   

18.
Abstract We examine the role of the native fox, Pseudalopex culpaeus, as a frugivore and seed disperser in a semiarid thornscrub of Chile. We quantified the fruit and animal components in its diet versus the availability of fruits and small mammals in the field over a 2‐year period (January 1998 through February 2000). We tested the legitimacy and effectiveness of foxes as dispersers by quantifying the percentages of seed viability and of germination of seeds that passed through fox gut versus those picked from plants. We also studied their efficiency as dispersers, monitoring the fate of seeds in faeces placed in the field. The highest frequencies of fruit consumption by foxes were observed when abundances of small mammal prey were <6 individuals per hectare, regardless of fruit abundance in the field. Thus, foxes consumed fruits as a supplementary food resource. Based on 326 faeces, the total number of fruits consumed was about 34 000 over the 2‐year study period, and fruits from the alien shrub Schinus molle represented 98% of that total, with the native Porlieria chilensis a distant second. Germination and viability of defecated seeds of P. chilensis were reduced by 66% and 48%, respectively, in comparison to controls. In contrast, germination of seeds of S. molle increased by 50% and no effect on viability was observed. With regard to P. chilensis, foxes were legitimate (they defecated viable seeds), but ineffective (seeds in faeces had lower germination than those taken directly from parental plants and there was no seedling establishment in the field) and inefficient dispersers (seeds in faeces were deposited on microhabitats hostile to seed germination and seedling establishment). However, with regard to S. molle, foxes were legitimate, effective (seeds in faeces had higher germination than those taken directly from parental plants; there was germination but no establishment in the field), and efficient dispersers (over 41% of seeds were deposited on safe microsites). Thus, a native fox may be contributing to the spread of an alien shrub, co‐opting existing community processes.  相似文献   

19.
兔狲(Otocolobus manul)、藏狐(Vulpes ferrilata)和赤狐(V. vulpes)是青藏高原三江源区域分布的重要小型食肉兽。本研究于2014年6月至2019年9月在青海省长江源区沱沱河和通天河沿岸选取208个位点布设红外相机, 通过所获取的时空分布数据比较了上述3种同域分布小型食肉兽的时空利用情况。通过空间重叠度系数的比较分析, 兔狲和藏狐、兔狲和赤狐以及藏狐和赤狐之间的空间重叠度系数分别为0.25、0.48和0.17, 这表明兔狲、藏狐和赤狐三者在空间利用上存在一定的差异。通过核密度估计方法分析, 兔狲和藏狐属典型的昼行性动物, 而赤狐以夜行性活动为主。兔狲、藏狐和赤狐每个物种在冷暖两季的日活动节律重叠指数分别为0.83、0.78和0.88。两两比较分析表明, 兔狲和藏狐二者的日活动节律重叠指数最高(0.84), 兔狲和赤狐在夜间活动时段存在一定重叠(0.63), 而藏狐和赤狐的时间生态位分化最明显, 重叠指数最低(0.48)。此外, 在暖季, 两两物种之间的日活动节律重叠指数均小于其冷季的重叠指数。综上所述, 长江源区兔狲、藏狐和赤狐3种小型食肉兽可通过空间和时间资源的利用差异来降低物种间的干扰和竞争, 从而达到同域物种共存的目的。  相似文献   

20.
Alveolar echinococcosis (AE) is a helminth zoonosis which is encountered only in the northern hemisphere. In central France, the Auvergne region represents the most western and southern extension of this helminthiasis. In 1999, a human case of AE was diagnosed in the southern part of the Cantal department, where AE was supposed absent, and an epidemiological survey was subsequently carried out. The transmission of the zoonosis in the sylvatic and peridomestic definitive hosts was studied, as well as that in the rodent and human intermediate hosts. Eleven red foxes (Vulpes vulpes) were shot, and 50 fox faecal deposits were collected. Twelve farm dogs had their faeces taken by rectal touch, and four were checked after arecoline purgation. Optical detection of Echinococcus multilocularis worms was achieved on fox intestines after scraping, and also on dog stools after arecoline therapy. Coproantigen ELISA assay was performed for the 11 scraping products, for the 50 fox faeces, and for the 12 dog faecal samples. No adult AE agent was observed by microscopy, and the ELISA assay yielded positive results in one of 11 fox intestines, one of 50 fox faeces, and 2 of 12 dog faecal samples. Twenty-five small mammals were trapped, of which 19 were Arvicola terrestris water voles. One rodent liver exhibited a hepatic lesion consistent with AE. An epidemiological questionnaire was completed in 85 human volunteers, who were also serologically tested for AE. Only one (the case's husband) exhibited a Western-blotting pattern indicative of a low-grade AE infection. The results of this preliminary study suggested a slow AE extension to the south of Cantal department from the northern focus.  相似文献   

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