The helminth fauna of red foxes (Vulpes vulpes Linnaeus 1758) in north Hesse and east Westphalia. 1. Cestodes]

Between November 1989 and June 1990 a total number of 397 foxes were examined for the presence of cestodes. The animals came from the districts of Arnsberg, Detmold and Kassel. In 16.4% of the foxes infections with Echinococcus multilocularis were found, in 28.5% Taenia crassiceps, in 14.4% Taenia polyacantha, in 4.3% Mesocestoides spp., in 3.3% Multiceps multiceps, in 2.5% Hydatigera taeniaeformis, in 0.8% Taenia hydatigena and in 0.3% Taenia martis. Infections with Echinococcus multilocularis very often showed high worm numbers of more than 1000 per fox. The number of worms for the other cestodes mostly varied between one and ten specimen per animal.     

Echinococcus multilocularis infections in domestic dogs and cats from Germany and other European countries

A cross-sectional survey was conducted to estimate the prevalence of Echinococcus multilocularis and E. granulosus infections in domestic dogs and cats from Germany and other European countries. Faecal samples of 21,588 dogs and 10,650 cats routinely submitted to a private veterinary laboratory between June 2004 and June 2005 were examined using the ZnSO(4)-NaCl flotation method. Taeniid eggs were detected in 54 (0.25%) and 37 (0.34%) of the canine and feline faecal samples, respectively. Taeniid eggs were separated and subjected to a DNA preparation and a modified two-step PCR for the detection of Echinococcus spp. based on mitochondrial 12S rRNA genes. PCR products from Echinococcus-negative but cestode-positive reactions were cloned and sequenced to determine the Taenia species. E. multilocularis DNA was specifically amplified in 43 (0.24%) and 25 (0.23%) of the samples from dogs and cats, respectively. E. granulosus DNA was not detected in any sample, while, E. multilocularis-positive samples were detected in dogs from Germany only, those of cats originated from Germany, Denmark and The Netherlands. The prevalence of E. multilocularis egg-positive canine samples was significantly higher in southern (0.35%) than in northern Germany (0.13%). In contrast, no significant regional difference was observed in cats from Germany. Taeniid eggs from Echinococcus-negative samples and from a few samples with macroscopically detected Taenia sp. proglottids were identified as eggs of T. crassiceps (n=8), T. martis, T. serialis, T. polyacantha, T. taeniaeformis and T. pisiformis in dogs (n=1 of each) and T. taeniaeformis (n=11) in cats. The spectrum of cestodes detected in domestic dogs and cats indicate the consumption of small rodents as infection source. The high proportion of E. multilocularis-positive samples, suggest domestic dogs and cats as a possible source of E. multilocularis infection for humans.     

Detection of Echinococcus multilocularis in foxes in The Netherlands

Echinococcus multilocularis was demonstrated in 5 out of 272 foxes in The Netherlands close to the border with Germany and Belgium. Besides microscopic examination of mucosal scrapings, two different PCR assays were used based on the detection of E. multilocularis DNA in colon content. Two distinct areas in The Netherlands were positive for E. multilocularis. Two positive foxes were found in the northern province of Groningen and three positive foxes were found in the southern province of Limburg. Both PCR assays detected more positive foxes compared to microscopic examination of the intestinal content. This is the first report of E. multilocularis in foxes occurring in The Netherlands.     

Echinococcus multilocularis: the prevalence of the fox tapeworm with possible serious consequences for humans

The prevalence of the fox tapeworm in foxes (final host) and muskrats (one of the intermediate hosts) in the Netherlands and Europe has been discussed. The tapeworm was found in 9.4% of the investigated foxes from the province of Groningen and in 0.2% of the muskrats from the same region. Also in the province of Limburg positive foxes were found, but no positive muskrats. Possible ways of infection for humans are described together with methods for prevention. It is concluded that at this moment risks for humans to become infected are minimal, but vigilance and monitoring of foxes and muskrats remains needed.     

Helminths of red foxes (Vulpes vulpes) in Denmark

An epidemiological study of helminths in 1040 red foxes collected from various localities in Denmark during 1997-2002, revealed 21 helminth species at autopsy, including nine nematode species: Capillaria plica (prevalence 80.5%), Capillaria aerophila (74.1%), Crenosoma vulpis (17.4%), Angiostrongylus vasorum (48.6% from Northern Zealand (endemic area)), Toxocara canis (59.4%), Toxascaris leonina (0.6%), Uncinaria stenocephala (68.6%), Ancylostoma caninum (0.6%), and Trichuris vulpis (0.5%); seven cestodes: Mesocestoides sp. (35.6%), a number of Taeniid species (Taenia pisiformis, T. hydatigena, T. taeniaeformis, T. crassiceps, and unidentified Taenia spp.) (22.8%), and Echinococcus multilocularis (0.3%); four trematodes: Alaria alata (15.4%), Cryptocotyle lingua (23.8%), Pseudamphystomum truncatum (3.6% from Northern Zealand), and Echinochasmus perfoliatus (2.4% from Northern Zealand); one acanthocephalan: Polymorphus sp. (1.2%). Significant difference in prevalence was found for T. canis and A. vasorum according to host sex, and for T. canis, U. stenocephala, Mesocestoides sp., Taenia spp., A. alata, A. vasorum, and Capillaria spp. according to age groups (adult, young or cub). Prevalence and average worm intensity for each helminth species varied considerably according to geographical locality, season, and year. Aggregated distribution was found for several helminth species. The two species E. multilocularis and E. perfoliatus are first records for Denmark.     

Hair coat contamination with zoonotic helminth eggs of farm and pet dogs and foxes

Infections of dogs with Toxocara canis and Echinococcus multilocularis pose an infection-risk particularly for contact persons. We examined specimens of hair coat and faeces of 124 farm dogs, 118 household dogs, 49 kennel dogs, 15 puppies from two litters, and 46 red foxes. Microscopically identified eggs of Toxocara or taeniids were further investigated by species-specific PCRs. In farm dogs, eggs of E. multilocularis or T. canis were identified in each 2.4% of faecal samples, eggs of T. cati (gastrointestinal passage) in 7.3%, respectively. Household dogs excreted eggs of T. canis (0.8%) and of T. cati (2.5%). In kennel dogs, eggs of T. canis (4.1%), but not of T. cati were detectable. Coat samples contaminated with eggs of Toxocara spp. were found from farm dogs (5.6%), household dogs (1.7%) and kennel dogs (2.0%). Taeniid eggs were isolated from the coat samples from only two farm dogs (1.6%); a molecular species determination was not achieved. In six intrauterinely infected puppies, Toxocara-eggs were found in 17/38 samples taken within six weeks. No intact Toxocara eggs could be isolated from the coat of nine puppies from a second litter 13 days after deworming. Of the 46 red foxes investigated (dissection and faecal samples) 13 (28.3%) were infected with E. multilocularis and 20 (43.5%) with Toxocara. Eggs of taeniids and Toxocara were found in 13% (in three cases confirmed as E. multilocularis) and 21.7%, respectively, of the coat samples. None of the retrieved Toxocara eggs in the coat samples were embryonated. Thus, an infection of humans through the transmission of E. multilocularis eggs after direct contact with dogs or foxes is conceivable, whereas a corresponding infection risk by Toxocara eggs must be critically challenged.     

Increase in the prevalence of Echinococcus multilocularis infection in red foxes in Lower Saxony.]

Echinococcus multilocularis is a tapeworm having carnivores as final hosts, the red fox in particular, dog and cat less frequent. Its two host life cycle consists of a larval cycle which predominantly takes place in the liver of rodents such as mice but it can also develop in musk rats as intermediate hosts. Man can also be infected and serves as a wrong intermediate host. He develops an alveolar echinococcosis which usually ends lethal without medical treatment. The prevalence of E. multilocularis among 5.365 red foxes in Lower Saxony was monitored from 1991 to 1997. The data were analysed using spatial epidemiological methods. This evaluation is based on a significance test which was applied to the parameters of spatial autoregressive regression models (CAR) fitting to the data of two successive sampling periods from 1991-1994 and 1994-1997. The mean prevalence (spatial median) increased from 6% to 11%. The results of this epidemiological study which was restricted to Lower Saxony support for the first time the earlier reported assumption that the prevalence of foxes infected with E. multilocularis has risen in Europe in the last decade. The reasons for this development are still unknown.     

Prevalence and intensity of Echinococcus multilocularis in red foxes (Vulpes vulpes schrencki) and raccoon dogs (Nyctereutes procyonoides albus) in Otaru City, Hokkaido, Japan

A survey was done in an attempt to investigate the epidemiological status of Echinococcus multilocularis in red foxes and raccoon dogs in Otaru city from June to September 1999. Sixty-seven red foxes (Vulpes vulpes schrencki) and 13 raccoon dogs (Nyctereutes procyonoides albus) were captured, and postmortem examinations were conducted with them. Thirty-eight red foxes (56.7%) and 3 raccoon dogs (23.1%) were found to be infected with E. multilocularis. The total biomass of E. multilocularis in all infected red foxes and raccoon dogs were 2,817,000 and 1,515 worms, respectively. Nine of the infected red foxes harboring more than 100,000 worms accounted for 90.6% of the total biomass. No significant differences in the prevalence were observed between male and female, and juvenile and adult. However, the worm burden was higher in juvenile than in adult foxes. In one of the infected raccoon dogs, mature worms and eggs of E. multilocularis were found in the intestine and fecal sample, respectively. This result suggested that the raccoon dogs are probably playing a small role in the egg contamination of the environment. The validity of coproantigen ELISA for diagnosis of foxes was confirmed by comparing the results of autopsy, egg examination and coproantigen ELISA using rectal fecal samples.     

The use of tetracycline in anthelmintic baits to assess baiting rate and drug efficacy against Echinococcus multilocularis in foxes

Anthelmintic (praziquantel) baiting of wild red foxes against Echinococcus multilocularis infection was studied in a highly epizootic suburban area of Otaru, Hokkaido (the northern island of Japan) during the summer and autumn in the years 1999-2004. Acceptance of baits containing the biomarker tetracycline (TC) was evaluated. The prevalence of E. multilocularis infection in foxes before baiting (1999-2000) was 58% (88/153), whereas in the fourth year of bait distribution year (2004), it decreased to 11% (5/45). Analysis of TC marking in the teeth of foxes showed that 39% (77/195) of those captured after baiting were estimated to have consumed baits in the year of capture. Importantly, more juvenile (56%, 49/87) than adult foxes (26%, 28/108) were marked, indicating efficient baiting of juveniles, which tended to have a higher worm burden of E. multilocularis. Of 77 marked foxes, E. multilocularis and Alaria alata (monitored as the second indicator species of deworming) were not detected in 70 (90%) and 76 (99%) foxes, respectively. The results suggest effective deworming by bait consumption. However, it was also demonstrated that 9% of the marked foxes were infected or re-infected after bait consumption, suggesting high infection pressure and the importance of frequent baiting.     

Serological (Em2-ELISA) and parasitological examinations of fox populations for Echinococcus multilocularis infections

Serum or body fluid samples of 1,006 foxes were investigated in an ELISA for antibodies against a highly sensitive and specific antigen (Em2-antigen) of Echinococcus multilocularis. Parasitological examinations of the intestines and simultaneous serological examinations were carried out in 505 foxes: A group of 98 blue foxes (Alopex lagopus) from Norwegian fox farms did not contain intestinal stages of E. multilocularis and was clearly sero-negative in Em2-ELISA. On the other hand in red foxes (Vulpes vulpes) originating from European areas known to be endemic for E. multilocularis the following average prevalence rates were found: 244 foxes from Southern Germany, E. multilocularis prevalence 55% and sero-prevalence 60%; 139 foxes from Austria, E. multilocularis prevalence 4% and sero-prevalence 12%. Serological identification of individual foxes with or without intestinal E. multilocularis infection was not possible. Only serological (no parasitological) examination in 402 foxes originating from endemic areas in Switzerland resulted in a sero-prevalence rate of 37%. Sero-prevalence was only 6% and 4% in 54 and 26 other foxes, respectively, originating from Swiss and German areas where E. multilocularis has not yet been reported. Negative control Norwegian (farmed) silver foxes (n = 43) were all sero-negative. The specificity of the Em2-ELISA was confirmed by negative Em2-serologies with sera from dogs infected with intestinal and tissue dwelling helminth species (with the exception of two from 24 dogs infected with E. granulosus).(ABSTRACT TRUNCATED AT 250 WORDS)     

Base line prevalence and spatial distribution of Echinococcus multilocularis in a newly recognized endemic area in the Netherlands

The base line prevalence of Echinococcus multilocularis in foxes was determined in the eastern part of the province of Groningen in the Netherlands adjacent to the German border. This region has been identified in a previous study in 1998 as one of the westernmost border areas of E. multilocularis. Base line prevalence data are important for a better insight in the possible spread of the parasite and its changes in time. As fox feces containing E. multilocularis eggs are an important source for human exposure this base line prevalence is also an indicator for the potential risk for public health. The base line prevalence was estimated at 9.4% (95% CI: 5.2-16.5%). These results confirm previous findings of E. multilocularis in the same region. The spatial distribution of the infected foxes has been analyzed as a spatial gradient using a logistic model. The prevalence appeared to change strongest in east-western direction and was highest near the German border, adjacent to a German endemic area. These results suggest that the border areas in the Netherlands are the most margin of E. multilocularis territory.     

Echinococcus multilocularis and Toxocara canis in urban red foxes (Vulpes vulpes) in Brussels, Belgium

During the last decades, European red foxes (Vulpes vulpes) have been implicated in the transmission of several viral or parasitic pathogenic agents to domestic animals and humans. In urban areas, risks of zoonoses transmission are likely to increase as a result of a higher rate of intra- and inter-species contacts. Foxes occur on 35% of the Brussels-Capital Region area and local densities reach up to 4 family groups per square kilometre. According to the directive 2003/99/ECC, a first survey for the presence in foxes of Echinococcus multilocularis and Toxocara canis was conducted in Brussels from 2001 to 2004. None of 160 foxes were found to be infected with E. multilocularis and 24 of 134 foxes were found to be infected with T. canis. Considering numbers of examined foxes, the sensitivity and the specificity of tests used for diagnosis, the 95% credibility intervals for the true prevalence of E. multilocularis and T. canis were estimated in a Bayesian framework to be 0 to 1.87% (median value of 0%) and 12.7 to 26% (median value of 18.7%), respectively. For T. canis, a significantly higher risk to be a carrier occurs in cubs and a significantly lower risk in adults.     

Echinococcus multilocularis (Cestoda,Taeniidae) in Red foxes (Vulpes vulpes) in northern Belgium

The first record of the tapeworm Echinococcus multilocularis (Cestoda, Taeniidae) in Red foxes (Vulpes vulpes) in northern Belgium is described. Between 1996 and 1999, 237 dead foxes were examined for the presence of this tapeworm using the intestinal scraping technique. Four foxes (1.7%) were found to be infected with E. multilocularis and showed medium to very high parasitic burdens. Three infected foxes originated from the south of the study area and the fourth animal came from the north of the study area near the border with The Netherlands. These findings are discussed in relation to the high endemicity of E. multilocularis in southern Belgium and to the increased distribution of the Red fox (V. vulpes) in northern Belgium during the last two decades.     

Echinococcus multilocularis in Grisons: distribution in foxes and presence of potential intermediate hosts

The southern border of the European endemic area of Echinococcus multilocularis runs along the alpine crest. This endemic transition area was analysed in the canton Grisons on a small spatial scale. A total of 543 foxes originating from 10 areas north and 4 areas south of the main alpine divide were investigated. Parasites were isolated using the sedimentation and counting technique on intestinal contents. The mean prevalence of E. multilocularis was 6.4% with significant differences between different areas. In the southern valleys, only foxes from the Val Müstair were infected (14.3%). On the northern side of the main alpine divide, prevalences varied between 0 and 40%. The predation habits of foxes on potential intermediate hosts was investigated by means of stomach content analyses (n=530). Rodents of the genera Microtus/Pitymys were found in 19.6% of fox stomachs, Clethrionomys glareolus in 8.0% and Arvicola terrestris in 0.4%. A small scale analysis based on a 10 x 10 km grid suggested that the predation rate on the genera Microtus/Pitymys significantly correlated with the prevalence of E. multilocularis in foxes (Spearman's r = 0.51). Hence, E. multilocularis appears to occur in the alpine study area on a very small spatial scale. These local sources of infection may persist for decades. This may partly explains why, during the last 34 years, some human cases of alveolar echinococcosis occurred in areas of the Canton Grisons where the parasite is presently endemic in foxes. No such human cases have been recorded in other areas free of E. multilocularis.     

Emergence of Echinococcus multilocularis among Red Foxes in northern Germany, 1991--2005

The parasite Echinococcus multilocularis is currently of great concern in Europe because of its spreading behavior, which has public health implications. This study investigates the spatiotemporal distribution of Echinococcus multilocularis in Red Foxes. The infection status of 8459 foxes was sampled from 43 regions in the north German province of Lower Saxony over three investigation periods 1991--1994, 1994--1997 and 2003--2005. Linear empirical Bayesian smoothing within the binomial model was used to produce smoothed choropleth maps. Geostatistical kriging was applied to generate prevalence risk maps. Further geostatistical modeling of the prevalence difference between study periods facilitated spatiotemporal trend investigations. The spatial scan statistic was used for cluster detection analysis. The average prevalence risk for Lower Saxony increased from about 12 to 20% during 1991--2005. Specifically the increases from first to second and to third study periods were estimated by 3.3% (CI 95%: 0.6%-5.9%) and 8.5% (CI 95%: 5.2%-11.8%), respectively. Infections in foxes were clustering and a location stable disease cluster was detected in the south of the province. This study is the first showing evidence for steady emergence of Echinococcus multilocularis in Red Foxes. First cases of human Alveolar Echinococcosis were recorded recently.     

Genetic uniformity of Echinococcus multilocularis collected from different intermediate host species in Hokkaido, Japan

DNA from several isolates of Taenia taeniaeformis and Echinococcus multilocularis were digested with restriction enzymes and hybridized with digoxigenated oligonucleotide probe (CAC)5. Within the six wild isolates of Taenia taeniaeformis from Norway rats in Hokkaido, although several bands were common among isolates, fingerprinting patterns were specific to each isolate. In the case of E. multilocularis, regardless of hosts from which each isolate has been isolated, the five isolates collected from Hokkaido, showed the same fingerprinting pattern. These results indicate that there was very little genetic difference among these isolates. Although the fingerprinting pattern of E. multilocularis from St. Lawrence Is. was similar to that of the Hokkaido isolates, some bands were different from those in the Hokkaido isolates. Echinococcus multilocularis in Hokkaido seems to be closely-related genetically to that from St. Lawrence Is.     

Prevalence of Echinococcus multilocularis in the red fox (Vulpes vulpes) in southern Belgium

Between June 1998 and February 2002, 709 red foxes killed in Wallonia (south of Belgium) were available for parasitological examination of the gut. The identification of Echinococcus multilocularis was based on morphological data. E. multilocularis adults were observed in 20.2% of the animals. The analysis of data revealed marked differences between the geological areas of Wallonia; the highest prevalence (33%) was found in the Ardenne and the lowest (0%) on the Plateau de Herve. Host gender and the collection season had no effect on the prevalence. However, the latter was significantly higher in juveniles (<8 months of age). The geographical distribution of E. multilocularis in Belgium is much wider than originally thought.     

Detection of Echinococcus multilocularis in foxes: evaluation of a protocol of the intestinal scraping technique

The intestinal scraping technique (IST) is widely used for the detection of definitive hosts infected with Echinococcus multilocularis. The sensitivity of the method has been questioned in recent years. Several variations of the technique are used that may differ in their performance. We therefore estimated the diagnostic sensitivity and specificity of the IST protocol used in our lab by examining the small intestines of 210 E. multilocularis-infected foxes and 294 foxes that had tested negative for this parasite. To this end, IST was first performed on 48 and 294 intestines from infected and uninfected foxes respectively, followed by an examination of the entire remaining mucosa of the small intestine to discover any infections with E. multilocularis that had not been detected by IST. The point estimate for diagnostic sensitivity was 100%. Five different classes of infection intensities were formed. The lower limits of the 95% confidence intervals ranged between 76.8% and 92.6% depending on the number of samples analysed per class. When the small intestines of another 162 infected foxes were examined by IST and the results recorded separately for the anterior, middle and posterior third of the small intestine, a strong preference of E. multilocularis for the posterior third was observed. For epidemiological purposes, it may thus be possible to restrict routine investigations to the posterior portion of the small intestine. By contrast, a reduction of the number of slides examined per infected animal may lead to a considerable loss in sensitivity in animals with a low intensity of infection.     

Helminth findings in indigenous raccoon dogs Nyctereutes procyonoides (Gray, 1843)

Internal organs of 74 racoon dogs originating from the eastern districts of the Federal Land Brandenburg were subjected to a complete helminthological dissection. In addition, samples of diaphragma muscles from 46 further animals were examined for Trichinella larvae. Altogether 3 trematode- (Alaria alata, Isthmiophora melis, Metorchis bilis), 3 cestode- (Mesocestoides sp., Echinococcus multilocularis, Taenia polyacantha) and 7 nematode species (Capillaria aerophila, C. plica, Trichinella spiralis, Uncinaria stenocephala, Molineus patens, Crenosoma vulpis, Toxocara canis) could be identified. Thereby, the racoon dog has a comparable helminth spectrum as well known in red foxes. Differences in the frequency of the occurrence of helminths in both hosts can be explained by different food preferences.     

Bayesian space-time analysis of Echinococcus multilocularis-infections in foxes

A total of 26,220 foxes that were hunted or found dead in Thuringia, Germany, between 1990 and 2009 were examined for infection with Echinococcus multilocularis, the causative agent of human alveolar echinococcosis, and 6853 animals were found infected. The available data on the foxes including the location (local community; district) and the date of hunting/death were analyzed using a hierarchical Bayesian space-time model. The distribution of the model parameters and their variability was estimated on the basis of the sample size, the number of cases per spatial unit and time interval, and an adjacency matrix of the municipalities using a Markov Chain Monte Carlo simulation technique to assess the spatial and temporal changes in the distribution of the parasite. The model used to evaluate the data is widely applicable and can be applied to analyse data sets with gaps and variable sample sizes per spatial and temporal unit. In the study area, the prevalence of E. multilocularis increased from 11.9% (95% confidence interval 9.9-14.0%) in 1990 to 42.0% (39.1-44.1%) in 2005. While the infection was present in foxes only in the north-western parts of Thuringia in 1990, it had spread over the entire state by 2004. These results demand increased vigilance for human alveolar echinococcosis in Thuringia.