[Efficacy evaluation of epsiprantel (Cestex) against Echinococcus mutilocularis in dogs and cats]. / Prüfung der Wirkung von Epsiprantel (Cestex) gegen Echinococcus multilocularis bei Hunden und Katzen.

Helminth-free dogs and cats were experimentally infected with protoscoleces of Echinococcus multilocularis and used in controlled trials for efficacy evaluation of the cestodicide epsiprantel. In two separate trials each 4 dogs were treated at day 20 post infection (p.i.) with average oral dosages of 5.1 (4.9-5.3) and 5.4 (5.2-5.8) mg/kg body weight (b.w.) epsiprantel, respectively, and necropsied at day 24 p.i. Among each 4 dogs of the two untreated control groups all animals were infected and had high intestinal worm burdens with averages of 33.575 and 100.725 E. multilocularis specimens per animal (individual worm burdens in group Ib 59,500-149,800, group IIb 20,500-43,200); in the two groups of treated dogs the average worm burdens were reduced by 99.6 and 99.9%. Among 8 treated dogs 4 were helminth-free, the other 4 had residual worm burdens (10-70 in 3 dogs, 1480 in 1 dog). In each 5 cats single oral treatments with average doses of 2.7 (2.7-2.8) and 5.5 (5.5-5.5) mg/kg b.w. epsiprantel were 100% effective against E. multilocularis 20 days p.i. and eliminated the worm burdens from all 10 animals. In the untreated group of 5 cats the average worm burden was 2864 per animal (individual worm burdens 20-6830). Side effects of the drug treatment were not observed. The results of the study show that in single therapeutic dosages recommended by the producer (dogs 5.5 mg, cats 2.75 mg/kg b.w.) epsiprantel eliminates E. multilocularis to over 99% or completely, but residual worm burdens may persist in some animals.

[Current spread and epidemiology of Echinococcus multilocularis]. / Zur aktuellen Verbreitung und Epidemiologie von Echinococcus multilocularis.

Recently, a wealth of new data was collected on the distribution and ecology of E. multilocularis. The parasite is now known to occur at surpisingly high prevalence rates in e.g. Belgium and northwestern Germany, new records exist for the Netherlands, and the parasite was found to be widespread in Poland and the Czech Republic. In addition, foxes in continental Europe have adapted their behaviour and are now common in many towns and cities where they are also known to carry the parasite. New data exist on endemicity regions in western Asia. In addition to new informations on the parasite's range, a summary is given of the current knowledge on prevalence of alveolar echinococcosis in man.

Experimental investigations on the B and T cell immune response in primary alveolar echinococcosis.

Susceptibility/resistance of the intermediate host to alveolar echinococcosis (AE) seems to be based on hitherto unknown immunological mechanisms, possibly involving the activation of different CD4+ T cell immune responses (Th1/Th2). Mice of two strains previously characterized as 'susceptible' (C57BL/6 J) and 'resistant' (C57BL/10 J) to secondary AE were orally infected with eggs of Echinococcus multilocularis and the course of infection was analysed by macroscopical, pathohistological and immunohistochemical examinations of the lymphocytes and cytokines participating in the periparasitic granulomas and by serological examinations of cytokines and E. multilocularis-specific antibodies. Although differences in the extent of parasitic growth were seen between the two groups, the composition of the granulomas was quite similar with CD4+ cells being the dominant lymphocyte subpopulation, succeeded by B cells and CD8+ cells. Interferon (IFN)-gamma-, interleukin (IL)-2- and IL-4-expressing cells could not be detected in the lesions of the early phase of the infection, possibly indicating the host's immunosuppression, but were present at the end. IL-10 was the most prominent cytokine throughout the course of the disease. Serological analyses of the cytokine concentrations revealed small amounts at the beginning and high levels at the end of the infection. The pattern of cytokine response was similar for IL-4 in both strains, but different for IL-2 and IL-10 in the late phase, when the C57BL/10 J strain developed higher levels than the C57BL/6 J strain. Correspondingly only small amounts of immunoglobulin (Ig)M, IgG1, IgG2a and IgG3 could be detected at the beginning of disease, followed by higher levels at the end. The courses of antibody titres were similar in both groups except IgG3, which was more pronounced in the C57BL/10 J strain. Parasite-specific IgG2b could neither be detected in the C57BL/6 J nor in the C57BL/10 J strain by the test system used. The results of the study suggest both subsets of CD4+ T cells (Th1 and Th2) being involved in murine primary alveolar echinococcosis. A strict differentiation of mice in susceptible and resistant animals based on the activation of different CD4+ T cell immune responses (Th1 'resistant' and Th2 'susceptible') should be avoided.

Detection of Echinococcus multilocularis in the definitive host: coprodiagnosis by PCR as an alternative to necropsy.

Recently, extensions of the range of Echinococcus multilocularis in Europe and North America and drastic increases in fox populations in Europe put an increasing proportion of the human population at risk of alveolar echinococcosis. To obtain data on the local infection pressure, studies of the prevalence of the parasite in the animals that transmit the parasite, foxes, dogs, and cats, are urgently required. Such investigations, however, have been hampered by the need for necropsy of the host animal to specifically diagnose infection with the parasite. In this study, a nested PCR and an improved method for DNA extraction were developed to allow the sensitive and specific diagnosis of E. multilocularis infections directly from diluted fecal samples from foxes. The target sequence for amplification is part of the E. multilocularis mitochondrial 12S rRNA gene. The specificity of the method was 100% when it was tested against 18 isolates (metacestodes and adult worms) of 11 cestode species, including E. granulosus. The sensitivity of the method was evaluated by adding egg suspensions and individual eggs to samples of diluted feces from uninfected foxes. The presence of one egg was sufficient to give a specific signal. To confirm the PCR results, an internal probe which hybridized only with E. multilocularis amplification products but not with the DNA of other cestodes was constructed. In order to investigate the applicability of this method for epidemiological studies, 250 wild foxes from a area in southern Germany where echinococcosis is highly endemic were examined by both necropsy and PCR of rectal contents. The sensitivity correlated with the parasites' number and stage of maturity. It ranged from 100% (>1,000 gravid worms) to 70% (<10 nongravid worms). On the basis of positive PCR results for 165 foxes, the sensitivity of the traditional and widely used necropsy method was found to be not higher than 76%. We therefore present this PCR system as an alternative method for the routine diagnosis of E. multilocularis in carnivores.

Echinococcus multilocularis in germany: increased awareness or spreading of a parasite?

The 'small fox tapeworm' Echinococcus multilocularis has recently become a matter of intense interest in Germany. A long-term increase of its prevalence in foxes has been noted in the well-known endemic areas in Southern Germany, and reports on the occurrence of the parasite in other parts of the country suggest that the parasite is actually much more widespread than previously thought. As nearly all of the relevant studies are published in the German language in a veterinary journal and in the hunting press, accessibility to the information is limited. Richard Lucius and Brigit Bilger here describe the situation, and discuss the possible reasons and consequences.

Influence of environmental factors on the infectivity of Echinococcus multilocularis eggs.

The sensitivity of eggs of Echinococcus multilocularis to environmental and controlled laboratory conditions was tested. Egg material was exposed and the infectivity was subsequently monitored by in vitro activation and by oral infection of the natural host, Microtus arvalis. To study the impact of environmental conditions in an endemic area of south-western Germany, eggs were sealed into bags of nylon mesh and exposed to the natural climate during various seasons. The maximal survival time of eggs was 240 days in an experiment performed in autumn and winter and 78 days in summer. A study of the tenacity of eggs under laboratory conditions revealed a high sensitivity to elevated temperatures and to desiccation. At 45 degrees C and 85-95% relative humidity the infectivity was lost after 3 h as well as after 4 h exposure to 43 degrees C suspended in water. Exposure to 27% relative humidity at 25 degrees C as well as exposure to 15% relative humidity at 43 degrees C resulted in a total loss of infectivity within 48 and 2 h, respectively. Temperatures of 4 degrees C and of -18 degrees C were well tolerated (478 days and 240 days survival, respectively), whereas exposure to -83 degrees C and to -196 degrees C quickly killed off the eggs (within 48 h and 20 h, respectively). Eggs of E. multilocularis were not killed off by exposure to various commercially available disinfectants applied according to the manufacturers' instructions and by exposure for 24 h to low concentrations of ethanol. Irradiation with 40 krad. from a 137Caesium source prevented the development of metacestodes but allowed seroconversion of infected rodents.